Note: Descriptions are shown in the official language in which they were submitted.
Therapeutic Modifiers of the Reverse Mode of ATP Svnthase
GRACE PERIOD
The inventor/applicant is also the inventor/applicant of PCT application
number
PCT/EP2018/051127 (filed 17 January 2018) and PCT application number
PCT/EP2018/069175 (filed 13 July 2018), which were published on 26 July 2018
and 17
January 2019 respectively, but they shoudn't be cited as prior art against the
present
disclosure because of the Canadian grace period.
FIELD OF THE INVENTION
This invention discloses compounds that preferentially slow the ATP-
hydrolysing mode of
ATP synthase, pharmaceutical compositions of these compounds, and methods of
use for
treating subjects known to have various diseases or disorders including cancer
(e.g. diagnosed
with), subjects suspected of having various diseases or disorders including
cancer or subjects
at risk of developing various diseases or disorders including cancer. In a
particular
embodiment, the subject is a human. In further embodiments, the subject is a
pet, or farm or
laboratory animal.
BACKGROUND OF THE INVENTION
ATP synthase
ATP synthase (also known as FiFo ATP synthase, FoFi ATP synthase, FiFo-ATPase,
FoFi-
ATPase, FIFO ATP hydrolase) is located at the inner mitochondrial membrane
(IM). It can
use the proton motive force (pmf) to generate ATP from ADP and Pi [1-3]. ATP
synthase is
reversible and - depending on its substrate/product concentrations, the pmf
and the voltage
across inner mitochondrial membrane {Tim} - it can work "forwards" (passaging
protons,
making ATP) or "backwards" (pumping protons, consuming ATP): its "forward" and
"reverse" modes respectively, which may also be termed FIF0ATP synthesis and
FiFo ATP
hydrolysis respectively.
Inhibitors of ATP synthase
There are drug inhibitors of ATP synthase, reviewed in [4] (herein
incorporated in its
entirety). Some inhibitors disproportionally/selectively inhibit the reverse
mode, as compared
to the forward mode, of ATP synthase [4-13]. Macrolides are a class of
polyketide. So
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CA 3050553 2019-07-25
macrolide FIFO ATP synthase inhibitors are polyketide FiFo ATP synthase
inhibitors, and
these terms are used interchangeably herein. Polyketide FIFO ATP synthase
inhibitors (e.g.
oligomycin) inhibit the forward mode, more than the reverse mode, of ATP
synthase [Ii].
Oligomycin is well known in the art as an inhibitor of FIR' ATP synthase, and
thence
oxidative phosphorylation and aerobic respiration [3]. Human life relies upon
aerobic
respiration. Indeed, the importance of breathing (02 in, CO2 out) is widely
appreciated.
Thence the danger of oligomycin is easily apparent.
IF1 is an endogenous protein, encoded by the ATPIF I gene, which selectively
blocks the
reverse mode of ATP synthase [4]. Its activity is pH sensitive and low, but
non-zero, at
normal matrix pH, and significant upon matrix acidification, caused by
collapse of the proton
motive force across the mitochondrial inner membrane.
Prior art teaches that compounds of this disclosure are NOT anti-cancer
therapeutics
Polyketide FiFo ATP synthase inhibitors (e.g. oligomycin) are poisonous to
cancer [14] and
normal [15] cells. Indeed, intraperitoneal injection of just 1 mg/kg
oligomycin kills healthy
rats (n=10) within 48 hours; LD33= 0.5 mg/kg [15]. Normal cells typically need
to use F1 F0
ATP synthase in its forward mode and so blocking this mode is typically
lethal. Thus,
polyketide FIFO ATP synthase inhibitors are not suitable as anti-cancer
therapeutics: indeed,
cytovaricin, ossamycin and peliomycin don't work in xenograft mouse models of
cancer
(data in [16], oligomycin untested) because a therapeutic window is absent
because, to repeat,
polyketide FiFo ATP synthase inhibitors are highly poisonous to normal cells,
whilst not even
being poisonous to all cancer cells: e.g. ineffective against glycolytic
cancers exhibiting the
Warburg effect [14]. [17] used oligomycin in a xenograft cancer mouse model
but only by
applying oligomycin to the cancer cells before they were inoculated into mice,
and washing
the excess oligomycin off before inoculation into the mice (by culture for 2
days in drug free
medium). They did the study like this (atypical, as clear to someone of the
art) because
oligomycin toxicity is not discriminate for cancer in a mammal. Obviously this
experiment
has no clinical parallel or utility. The synthesis/structure of some molecules
of this disclosure
has been disclosed in prior disclosures [PI, P2. P3], wherein these structures
are speculated to
be anti-cancer medicines merely by analogy to the anti-cancer activity of
polyketide FiFo
ATP synthase inhibitors in [14]. Indeed, to mirror and use the restriction of
[14], these
disclosures restrict their suggestion to "cancers having tumor cells that do
not exhibit the
Warburg effect" (in a claim of [P3], [Pl] doesn't claim for any cancer, [P2]
cancer claim
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rejected by USPTO in correspondance on 4/11/2006). [P1, P2, P3] state in the
Utility section
of their Description, "inhibitors of mitochondrial FiFo-ATPase selectively
kill metabolically
active tumor cells that do not exhibit the Warburg effect i.e., cells that do
not maintain a high
level of anaerobic carbon metabolism even in the presence of oxygen". So,
teaching that their
compounds will not exert anti-cancer activity against cancers that exhibit the
Warburg effect
i.e. they restrict their suggestion to cancers using oxidative phosphorylation
(OXPHOS) and
ATP synthase, in its forward mode, to generate ATP. But what undermines this
(postulated)
approach is that this aerobic profile is what normal cells typically use also,
especially on
aggregate across an organism: well known to those of the art (evidence:
importance of
breathing to mammalian life). By this analogy to polyketide FIFO ATP synthase
inhibitors,
these disclosures speculate these molecules are safe anti-cancer therapeutics.
When in fact, by
this analogy, they actually teach the opposite. This is clear when [14] isn't
considered in
isolation, as it shouldn't be, but alongside the rest of the literature e.g.
[15], [16] and the
knowledge of someone of the art (well known that normal cells need to use FiFo
ATP
.. synthase in its forward mode, to generate ATP, and that oligomycin blocks
this, and is
potently dangerous). So, these prior disclosures [P1, P2, P3] teach someone of
the art, that
these compounds are, by their chosen analogy to polyketide FIFO ATP synthase
inhibitors,
not suitable for anti-cancer therapy. It isn't sufficient to kill cancer to be
an anti-cancer
therapeutic. This killing must be selective, leaving normal cells alive.
Metabolic poisons such
.. as cyanide or oligomycin do not fit this criterion. By distinction, the
present invention
discloses selective killing of cancer cells, at compound concentrations
harmless to normal
cells. This couldn't have been anticipated from the prior art. Furthermore,
this selective anti-
cancer activity is pronounced for cancers that do exhibit the Warburg effect.
Distinctly, the present disclosure discloses experimental data. Its inventive
step is to show
that its compounds are safe anti-cancer therapeutics, exactly because of their
distinction from
polyketide FiFo ATP synthase inhibitors. There is a broad therapeutic margin
for the
compounds of this disclosure as a virtue of the distinctive (from oligomycin)
way they work,
leveraging differences between normal and cancer cells, discovered and
disclosed as part of
.. this invention. Indeed, the compounds of this disclosure can kill highly
glycolytic cancers
exhibiting the Warburg effect. These cancers tend to be the most dangerous,
with the worst
prognosis (numerous studies find this: representatives: [18-20]).
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CA 3050553 2019-07-25
[Pl, P2, P3] do not provide any experimental evidence of any anti-cancer
activity, and teach
towards treating ischemia. They teach away from the present invention by
suggesting, by an
analogy in the unpredictable arts, that their compounds exert anti-cancer
activity, like
oligomycin, by inhibition of FIR' ATP synthesis, wherein oligomycin itself is
unsuitable as
an anti-cancer drug. [Pl, P2, P3] contains millions of compounds. The person
of the art
would select a subset of these for anti-cancer testing. Teaching of [P1, P2,
P3] teaches the
person of the art to select compounds that maximally inhibit FiFo ATP
synthesis in the
submitochondrial assay they describe. Such compounds would very potently kill
cancer in
vitro, like oligomycin in [14], and so be excitingly entered into subsequent
animal studies,
wherein a lack of therapeutic margin would become evident (like polyketide Fi
Fo inhibitors
in [16]) and no useful cancer drugs would be found, concluding the
experimentation. Indeed,
none found in subsequent ¨16 years, despite long standing need. Whereas, by
the present
invention, compounds are selected from [Pl, P2, P3] for anti-cancer activity
by the inverse.
By selection of compounds that minimally inhibit FIFO ATP synthesis (as
compared to their
inhibition of Fi Fo ATP hydrolysis). Indeed, by experimental evidence herein,
the anti-cancer
activity of such compounds is evidential. Thus, enabling the person of the art
to arrive at
working cancer drugs, and having the rationale to arrive at further working
cancer drugs e.g.
found by a method(s) disclosed herein. These drugs can cause body temperature
drop, and
can have a very atypical dose-anticancer response profile, which are critical
teachings, with
compensatory and associated methods, of this invention. These teachings are
enabling for
rodent trials, and more especially mouse trials (small body, more susceptible
to body
temperature drop), which is a step that one of the art would use to assess
compounds of this
invention.
Compounds of the present invention don't just exert anti-cancer activity. They
can also affect
normal cells, making their metabolism more efficient, which can cause weight
gain/reduce
weight loss/maintain body weight, all of which combats cachexia. For example,
cancer driven
cachexia, which is the leading cause of death in cancer patients. By contrast,
polyketide Fi Fo
ATP synthase inhibitors are toxic to normal cells, denying them energy, rather
than enabling
them more energy, by efficiency gain, as molecules of this invention can do.
In short, [P1, P2, P3] teach that FIR ATP synthesis inhibitors only kill
oxidative cancers not
using Warburg metabolism, this invention experimentally shows that FIR ATP
hydrolysis
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inhibitors do kill cancers using Warburg metabolism. The former teaching
doesn't arrive at
therapeutic cancer drugs, the latter does.
[P4] teaches away from the present invention. Its exemplary compound, Bz-423,
inhibits the
forward and reverse modes of ATP synthase equally or, in other data it
discloses, the forward
(EC50= 5.5 M) more than the reverse (ECK' = 8.9 M) mode of ATP synthase.
[P4]
provides arguments and evidence that Bz-423 slows cellular proliferation, and
causes
apoptosis, by inhibiting FIR' ATP synthesis and not by inhibiting FIFO ATP
hydrolysis. It
extrapolates this feature of Bz-423 to other compounds it discloses (a point
the (same)
.. applicant of [P5] stresses often in examination correspondence with the
USPTO). For
example, in the title of Example 41 in [P4], it is written "Benzodione
derivatives inhibit ATP
hydrolysis, does not affect cell synthesis properties, and does not affect
cell viability", and
[P4] goes on to affirm this statement with experimental data, which teaches
away from the
present invention, wherein compound T5 is "capable of inhibiting ATP
hydrolysis, not
.. inhibiting cell synthesis, not affecting cell viability". In [P4], T5
experimental data is
included to contrast with that of Bz-423, to show the lack of utility of a
specific ATP
hydrolysis inhibitor such as T5, as compared to the utility of the exemplary
compound, Bz-
423, which inhibits ATP synthesis. In this data in [P4], Bz-423 inhibits FiFo
ATP synthesis
and hydrolysis and cellular viability, T5 only inhibits FIR ATP hydrolysis and
not cellular
viability, = Bz-423 is the exemplary compound and FIR ATP synthesis
inhibition, not FiFo
ATP hydrolysis inhibition, is experimentally highlighted as the responsible,
exemplary
mechanism. Bz-423 hyperpolarises Tim and decreases 02 consumption [21] whilst
compounds of the present invention do not, as presented in experimental data
of the present
disclosure. Example 52 in [P4] states "For ATP synthesis (the relevant
enzymatic reaction of
the mitochondrial FiFo-ATPase in vivo)". Thence stating that FiFo ATP
hydrolysis is
irrelevant in vivo, which the present invention discloses, with supporting
experimental data, is
a falsehood. This is a new fundamental biological discovery, which will
surprise those of the
art. [P4] teaches the use of inhibitors of the forward mode of ATP synthase,
which is not
teaching that arrives at the present invention; indeed, it teaches away from
the present
invention.
Well known to those of the art, biorxiv is a repository for documents that
have not been peer
reviewed. In 2015, in a document on biorxiv, the author of this present
disclosure suggested
the use of FIFO ATP hydrolysis inhibitors as anti-cancer therapeutics [22].
This document was
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subsequently submitted to peer reviewed journals (elife, BMC Cancer) and was
found
unworthy of dissemination, let alone pursuit, by those of the art. It
demonstrably wasn't
considered credible by those of the art. In 2017, a paper was published by
others, in a peer-
reviewed journal [23], with experimental data showing that inhibiting h Fo ATP
hydrolysis
assists (!), rather than harms, cancer. With this conclusion reached and
emphasised by its
authors. This paper is one of many experimental reports, published in leading
peer reviewed
journals, reaching the same conclusion, which directly opposes and teaches
away from this
author's suggestion in a document database well known for
unreviewed/unscrutinised
manuscripts. Indeed, no peer-reviewed journal publication years after a
biorxiv submission
would be noted very negatively by one of the art.
When selecting a path to pursue from the prior art, in the unpreditable arts,
one of the art will
always weigh experimental data more heavily than suggestion. Especially
wherein
experimental data postdates and falsifies/discredits suggestion. Experimental
data in the prior
art teaches away from the present invention. Indeed, it directly opposes it.
Whereas the
present invention is of FiFo ATP hydrolysis inhibition conveying anti-cancer
therapy,
experimental data in [23] shows that FIFO ATP hydrolysis inhibition assists
cancer (!), thence
increasing its danger. "The ATP synthase complex does not hydrolyze ATP in
either
IF i-expressing or Ih-silenced osteosarcoma cells" [23]. "Even severe hypoxia
could not
activate the hydrolysis of ATP by the FiFo-ATPase complex" [23]. "ATP synthase
does not
hydrolyze ATP in cancer cells" [23]. Moreover, "in cancer cells IF)
overexpression fully
prevents ATP synthase hydrolytic activity" and "IF' is present at higher
levels in cancer cells
than in untransformed cells" (also observable in gene expression databases,
also reported by
many other investigators in many other journal papers, lh overexpression in
cancer is a
prognostic marker of poor patient outcome, IF) knockdown suppresses tumour
growth in
mice [241) and "lh overexpression promotes cancer cells survival" [23].
In Claim 1 of this disclosure, some compounds are excluded from the presented
formula by
proviso, separating all its enumerations from [P1] and [P4].
Some guidance
All publications, patents and patent applications mentioned or cited in this
disclosure are
herein incorporated, in entirety, by reference. This disclosure uses ICso and
ECso
interchangeably, for a process being inhibited or reduced. Chemical structures
were drawn
6
CA 3050553 2019-07-25
using the chemical drawing feature in [25], and if a drawing feature is
unknown to the reader
they are referred to its documentation, or to explore the software themselves:
all clear to those
of the art. Hydrogen on structures is typically not shown, present implicitly,
but it is shown
for some presented structures "On Hetero and Terminal" [25] groups. Herein,
the symbol D is
used for deuterium (2H). For compound synthesis schemes herein, starting
materials are
commercially available or can be readily prepared by one of ordinary skill in
the art using
known methods or derived by procedures analogous to those described in the
literature.
Examples and preparations herein describe the manner and process of making and
using the
invention. It should be understood that there will be other embodiments which
fall within the
spirit and scope of the invention. A phrase in the form "A/B" or in the form
"A and/or B"
means (A), (B), or (A and B).
SUMMARY OF THE INVENTION
The compounds described herein, including for example almitrine dimesylate,
exert anti-
cancer activity because they reduce FIFO ATP hydrolysis in cancer cells. This
elucidated
mechanism is the core of this invention. Herein is the discovery of a cancer-
specific drug
target: the reverse mode of ATP synthase. Indeed, new experimental data,
disclosed herein,
demonstrates that molecules which specifically slow FIFO ATP hydrolysis can
exert specific
anti-cancer activity, at concentrations that do not harm normal cells. Any
anti-cancer drug
that targets/inhibits/reduces FiFo ATP hydrolysis is componentry to this
invention. This
disclosure discloses numerous anti-cancer drug working examples, many of which
are also
new compositions of matter, and discloses rationale and methods to find
further working
examples, which are, in turn, componentry to this invention and encompassed by
this
disclosure.
Almitrine dimesylate is 644-[bis(4-fluorophenyl)methyl]piperazin-1-y1]-2-N,4-N-
bis(prop-2-
enyl)-1,3,5-triazine-2,4-diamine methanesulfonic acid, the dimethanesulfonate
salt of
almitrine. Almitrine CAS number is 27469-53-0 and almitrine has the following
structure:
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N
N N
H2C I I
Almitrine dimesylate has been used clinically for millions of patient months
to treat chronic
obstructive pulmonary disease (COPD). However, after being used for decades,
it is now only
used sporadically. Because postmarketing surveillance has revealed it doesn't
actually treat
COPD, thence there is no reward to outweigh its side-effects risk, especially
since there are
better/working COPD treatments. Although, for example, almitrine is still in
the formulary of
the National Health Service (NHS) of Great Britain for its hospital/specialist
doctors to
prescribe for respiratory disorder. This invention repurposes almitrine for
anti-cancer
treatment. New experimental data herein shows that almitrine dimesylate exerts
strong anti-
cancer activity, greater than carboplatin in standardised NCI-60 testing at
the National Cancer
Institute (NCI, USA). Carboplatin is one of the most used chemotherapies today
and is on the
World Health Organisation (WHO) list of essential medicines, but carboplatin
has a terrible
side-effect profile. By contrast, the potential (don't occur in most patients)
side-effects of
almitrine are mild compared to present chemotherapies, and tend to only occur
with chronic
use, yielding the opportunity for a therapeutic window of treatment, which
will
treat/ameliorate/prevent/combat cancer in a subject.
An aspect of the invention is a compound, or a composition containing at least
one
compound, of the following formula:
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RB
I
........-N,...õ
N/
)"----..
N - N
R )1..... ........;:i,
Ai ....,
N N NH
H I
R A2
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
for use in a method
of treating, ameliorating, preventing or combating cancer, or cancer that
metabolizes much of
its glucose and/or glutamine to lactate, for example a cancer exhibiting the
Warburg effect
and/or a cancer that can be discriminated from surrounding tissue by PET
imaging (e.g. 18F-
FDG PET); wherein:
RAI and le2 are each independently selected from the groups
and
wherein Rc and RD are each independently selected from hydrogen, deuterium,
halogen and
alkyl, and wherein RE is hydrogen, deuterium, or alkyl;
RB is selected from RBI, hydrogen and deuterium;
wherein RBI is selected from phenyl, benzyl, pyridyl, pyrimidyl and pyrazinyl
optionally substituted with one or more substituents RB2;
wherein each RB2 is independently selected from halogen, alkyl, alkoxy, nitro,
amino,
methoxy and polyhalogen alkyl;
or RB is a phenylalkyl of the formula:
9
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11 Gq (RG)q
(RF) ¨I. )n 1
wherein le and RG are hydrogen or alkyl, G is a carbon-carbon double bond or a
carbon-
carbon single bond, n is 0 or 1 and q is 0 or 1 provided that where q is 0, G
is a carbon-carbon
double bond and where q is 1, G is a carbon-carbon single bond,
or RB is a diphenylalkyl of the formula
RH
_
= P
RH
wherein RH is hydrogen or halogen, and p is 0, 1 or 2;
or RB is the group
CA 3050553 2019-07-25
Rj
RK
wherein RJ and RK each independently represent 1-5 optional substituents on
each ring, and
wherein each It! and each RK, when present, is independently selected from
halogen, alkyl,
alkoxy, nitro, amino, methoxy and polyhalogen alkyl.
In some embodiments, RB is the group:
RJ
RK
In some embodiments, RB is the group:
11
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Rj
e
it
RK
wherein IV and RK each independently represent 1 or 2 substituents on each
ring, and wherein
each RI and each RK is independently selected from halogen, alkyl, alkoxy,
nitro, amino and
polyhalogen alkyl.
In some embodiments, le and RK each independently represent 1 or 2
substituents on each
ring, and wherein each IV and each RK is independently selected from halogen.
In some embodiments, IV and RK each independently represent I substituent on
each ring,
and wherein IV and RK are independently selected from halogen.
In some embodiments, RB is the group:
RL
RM
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wherein le- and Rm are each independently selected from halogen, alkyl,
alkoxy, nitro, amino
and polyhalogen alkyl.
In some embodiments, RL and Rm are each independently selected from halogen.
In some embodiments, RL and Rm are the same.
In some embodiments, RL and Rm are each F.
In some embodiments, RAI and RA2 are each independently selected from the
group
RD
wherein Rc and RD are each independently selected from hydrogen, deuterium,
halogen and
alkyl.
In some embodiments, RAI and RA2 are the same.
In some embodiments, Rc is hydrogen. In some embodiments, RD is hydrogen. In
some
embodiments, Rc and RD are the same. In some embodiments, Rc and RD are both
hydrogen.
In some embodiments, the compound is:
F F
"-..N./
N - N
N N N
H H
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof.
In some embodiments, the compound is an isotopologue(s) of:
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F F
,....--N-...,
\N---'
NIN..- N
H2C..,=== A., 1õ ,.,-.....,,...,-. CH2
N N N
H H
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof.
Also described herein for prevention or treatment of cancer in a subject,
particularly with
cancer exhibiting the Warburg effect, is to use a pharmaceutical composition
with an
effective amount of one or compounds of the following formula,
Z R2 R3N.N.,
N/s-NN= N
L H
I
(R4) R1
q
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
L is alkyl, or substituted alkyl, or deuterated alkyl, or aminoalkyl, or
thioalkyl, or
alkoxy, or halogen, or haloalkyl, or haloalkoxy, or hydroxyalkyl, or any atom
or isotope
permitted by valence (including any accompanying hydrogens by valence e.g.
(non-limiting)
OH, NH2, SH, SiH3, PH2 etc.);
RI is hydrogen, cyano, ¨S02R8, ¨C(=0)R9, heteroaryl or thiazolyl;
R2 is (i) independently hydrogen, alkyl, benzyl, or substituted alkyl, or (ii)
taken
together with R3 forms a heterocyclo;
R3 is (i) independently alkyl, substituted alkyl, alkylthio, aminoalkyl,
carbamyl, BB-
aryl, BB-heterocyclo, BB-heteroaryl, or BB-cycloalkyl, or (ii) phenyl
optionally substituted
with Cmalkyl, halogen, trifluoromethyl, OCF3, cyano, nitro, amino, hydroxy, or
methoxy, or
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CA 3050553 2019-07-25
(iii) independently selected from Ci_aalkyl, alkylthio, aminoalkyl, -BB-aryl, -
BB-heterocyclo,
BB-cycloalkyl, and -BB-hetaroaryl, optionally having one to three substituents
selected from
R3a; and/or having fused thereto a five or six membered carbocyclic ring, or
(iv) taken
together with R, forms a heterocyclo optionally substituted with alkyl or
substituted alkyl;
BB is a bond, Ci_aalkylene, C2_4alkenylene, substituted Chaalkylene,
substituted C2-
4a1keny1ene, substituted Ci_aalkylene-C(=D)NH-, -C(=3)NH-, -C1.4alkylene-
C(=0)NH-, -C(=0)NR19-, -C malkylene-C(=a)NR19-, or substituted C1_4alkylene-
C(=0)NR19-, -(CHR14)m-(CRI5R16),- or -(CHR14)1,--C(=0)NH-;
R3a at each occurrence is selected independently from alkyl, substituted
alkyl,
halogen, haloalkoxy, cyano, nitro, keto, trifluoromethyl, -NRI7R18, -SR17, -
011.17, -
SO2R17a, -SO2NR17R18, -NR17COR18, -0O2R17, -C(D)R17, cyoloalkyl, aryl,
heterocyolo, and heteroaryl, wherein when R3a is cycloalkyl, aryl, heterocyclo
or heteroaryl,
said cycloalkyl, aryl, heterocyolo and heteroaryl in turn is optionally
substituted with alkyl or
substituted alkyl;
Z is a heteroaryl, for example an optionally-substituted bicyclic heteroaryl;
or
Z is triazolyl optionally substituted with one to two R7 substituents or
imidazolyl
optionally substituted with one to two R7 substituents and/or having fused
thereto a benzene
ring in turn optionally substituted with one to two R7 substituents; and
R7 is alkyl, carbamyl, or substituted alkyl;
R4 at each occurrence is selected independently of each other R4 from the
group
consisting of halogen, trifluoromethyl, OCF3, alkyl, substituted alkyl,
haloalkyl, nitro, cyano,
haloalkoxy, 0R25, SR25, NR25R26, NR25S02R27, S02R27, S02NR25R26, CO2R26,
C(=0)R26,
C(=0)NR25R26, OC(=0)R25, -0C(=0)NR25R26, NR25C(=D)R26, NR25CO2R26, aryl,
heteroaryl, heterocyclo and cycloalkyl;
R8 is Cmalkyl or phenyl optionally substituted with alkyl, halogen,
haloalkoxy, cyano,
nitro, or trifluoromethyl;
R9 is -NR10R11, alkyl, substituted alkyl, alkoxy, alkylthio, cycloalkyl, aryl,
heteroaryl, heterocyclo, or -0O2R12, alkyl or phenyl optionally substituted
with one to four
of halogen, cyano, trifluoromethyl, nitro, hydroxy, Chaalkoxy, haloalkoxy,
Ci_6alkyl,
CO2alkyl, SO2alkyl, SO2NH2, amino, NH(Ci_aalkyl), N(Ci_4alky1)2, NHC(=0)alky,
CA 3050553 2019-07-25
C(=0)alkyl, and/or Ci_aalkyl optionally substituted with one to three of
trifluoromethyl,
hydroxy, cyano, phenyl, pyridinyl; and/or a five or six membered heteroaryl or
heterocylo in
turn optionally substituted with keto or having a benzene ring fused thereto
or
a) Ci_aalkyl optionally substituted with one to two of:
i) SR13, 0R13, NRI3aRi3b, halogen, trifluoromethyl, CO2R13a, and
C(=0)NIZI3aR13b;
ii) cycloalkyl optionally substituted with one to two of C(=0)H, Cmacyl,
alkenyl, carbamyl,
and/or phenyl in turn optionally substituted with halogen;
iii) phenyl or napthyl optionally substituted with one to two of halogen,
nitro, amino, alkyl,
hydroxy, Ci_aalkoxy, or having fused thereto a five or six membered
heterocyclo;
iv) pyridinyl, thiophenyl, furanyl, tetrahydrofuranyl, or azepinyl, optionally
substituted with
alkyl or having fused thereto a five to six membered carbocyclic ring
optionally substituted
with keto or Ci_aalkoxy;
b) 3 to 6 membered cycloalkyl optionally having up to four substituetits
selected from alkyl,
halogen, cyano, alkenyl, acyl, alkylthio, carbamyl, phenyl in turn optionally
substituted with
halogen; or having an aryl fused thereto;
c) pheyl optionally substituted with one to four of halogen, cyano,
trifluoromethyl, nitro,
hydroxy, Ci_aalkoxy, haloalkoxy, Ci_aalkyl, CO2alkyl, SO2alkyl, SO2NH2, amino
NH(C3-
4a1ky1), N(Ci_4alky1)2, NHC()alkyl, C(=C0)alkyl, and/or Ci_aalkyl optionally
substituted
with one to three of trifluoromethyl, hydroxy, cyano, phenyl, pyridinyl;
and/or a five or six
membejed heteroaryl or heterocyle in turn optionally substituted with keto or
having a
benzene ring fused thereto;
d) pyridinyl, thiazolyl, furanyl, thiophenyl, and pyrrolyl optionally
substituted with one to
two of halogen, alkyl, and phenyl in turn optionally substituted with halogen
or
trifluoromethyl;
Rio and RI I are (i) independently selected from hydrogen, alkyl, substituted
alkyl,
alkoxy, heterocyclo, cycloalkyl, aryl, heteroaryl or Ci_aalkyl optionally
substituted with one
to two of ¨0O2alkyl, ¨C(=0)NH(ary1), NH(ary1), cycloalkyl, phenyloxy, phenyl
in turn
optionally substituted with Ci_aalkyl, hydroxy, Ci_aalkoxy, halogen, amino,
nitro,
tetrahydrofuranyl, and/or five or six membered heterocyclo, or having a five
or six membered
16
CA 3050553 2019-07-25
heterocyclo fused thereto; pyrrolidinyl optionally substituted with keto;
napthyl, anthracenyl,
pyridinyl, thiophenyl, furanyl, imidazolyl, benzimidazolyl, or indolyl in turn
optionally
substituted with Ci_aalkyl or Ci_4alkoxy; or (ii) taken together form a
heteroaryl or
heterocyclo selected from pyrrolidinyl, piperazinyl, piperidinyl, morpholinyl,
tetrahydropyridinyl, and imidazoilidinyl, wherein said heterocyclo formed by
Rio and Ri i is
optionally substituted with one to two of keto, CO2H, Ciaalkoxy, CO2alkyl,
Ci_acarbamyl,
benzyl; phenyl in turn optionally substituted with alkyl, halogen, or C1-4
alkoxy;
tetrahydropyridinyl in turn optionally substituted with keto and/or phenyl;
alkyl optionally
substituted with amino or NHR21wherein R21 is alkyl or phenyl optionally
substituted with
alkyl; and/or has a benzene ring fused thereto in turn optionally substituted
with one to two of
alkyl, Ci_aalkoxy, CO2alkyl, and/or Ci_acarbamyl;
R12 and R19 are hydrogen or alkyl;
R13 is hydrogen or alkyl;
Ri3a and R13b are selected from hydrogen, alkyl, and aryl;
R14, R15 and R16 at each occurrence are independently selected from hydrogen,
alkyl,
hydroxy, hydroxyCi_aalkyl, Ci_aalkoxy, and phenyl, and/or one of R15 and one
of Rio join
together to form a 3 to 6 membered cycloalkyl;
R17 and R18 are independently selected from hydrogen, alkyl, substituted
alkyl, aryl,
phenyl, or benzyl wherein the phenyl or benzyl is optionally substituted with
alkyl, hydroxy,
or hydroxyalkyl;
RI% is alkyl or substituted alkyl;
R25 and R26 are independently selected from hydrogen, alkyl, or substituted
alkyl, or
taken together form a heterocyclo or heteroaryl ring;
R27 is alkyl or substituted alkyl;
q is 0, 1, 2, or 3;
m and n are 0, 1 or 2; and
p is 0, 1, 2, or 3.
17
CA 3050553 2019-07-25
In some embodiments, there is an enantiomeric excess of the enantiomer:
R2 R3
NN/
oµµµL
Id1N.'"/%N
HI
(R4)q R1
Herein, the terms "S-stereoisomer" and "S-enantiomer" refer to the arrangement
of groups
around the chiral centre shown in the structure above, regardless of the
specific identities of
the variables such as Z, L and R4 within the structure. This chiral
configuration is termed S by
IUPAC designation if Z is nitrogen and L is deuterium for example. But,
herein, where S is
written next to this configuration it is not intended to limit what atoms can
be Z or L
according to IUPAC stereoisomer naming rules. For example, herein, Z can be
carbon (or
nitrogen or other atoms as specified herein) even when an S is written next to
this chiral
carbon. However, when R group type aliases, such as Z and L, are not used and
all atoms are
uniquely specified around a chiral carbon then IUPAC stereoisomer naming is
adhered to.
In some embodiments, the S-enantiomer of the compound is in enantiomeric
excess. In some
embodiments, the enantiomeric excess of 5-enantiomer exceeds 70%.
In some embodiments, L is hydrogen or deuterium.
In some embodiments, L is alkyl, or deuterium, or substituted alkyl, or
deuterated alkyl, or
aminoalkyl, or thioalkyl, or alkoxy, or halogen, or haloalkyl, or haloalkoxy,
or hydroxyalkyl,
or any atom or isotope permitted by valence except hydrogen at natural
abundance.
In some embodiments, the compound is a compound according to the formula
R2 R3
NN/
N/k.N*. N
H H
(R4) R1
q
18
CA 3050553 2019-07-25
or pharmaceutically-acceptable salts, solvates, hydrates or prodrugs thereof.
In some embodiments, the compound is a compound according to the formula
R2 R3
00H
S
Nrk'N
R1
(R4)q
or pharmaceutically-acceptable salts, solvates, hydrates and prodrugs thereof,
wherein S symbolises the S stereoisomer, for example, in enantiomeric excess
(ee) exceeding
70%.
In some embodiments, the compound is a compound according to the formula
R2N3
R.
/j\NN
N N
D H
R1
(R4)q
or pharmaceutically-acceptable salts, solvates, hydrates and prodrugs thereof,
wherein D is deuterium (enrichment, for example, exceeding 40% deuterium
incorporation at
shown position, and optionally at other positions also).
In some embodiments, the compound is a compound according to the formula
R2
NR 3
ttoDzIN
S NH N\ rii
R1
(R4)q
19
CA 3050553 2019-07-25
or pharmaceutically-acceptable salts, solvates, hydrates and prodrugs thereof,
wherein D is deuterium (enrichment, for example, exceeding 40% deuterium
incorporation at
shown position, and optionally at other positions also);
S symbolises the S stereoisomer, for example, in enantiomeric excess (ee)
exceeding 70%.
In some embodiments, the compound is a compound according to the formula
R2 R3
Z N.N,
N//kNN
CH3 H I
(R4) R1q
or pharmaceutically-acceptable salts, solvates, hydrates or prodrugs thereof.
With a
compound of this formula, in some embodiments, there is an enantiomeric excess
of
enantiomer. In some embodiments, the S-enantiomer of the compound is in
enantiomeric
excess. In other embodiments, the R-enantiomer of the compound is in
enantiomeric excess.
To illustrate, for (non-limiting) example, with supporting experimental data
herein, against
some cancers, the S-enantiomer exerts more potent anti-cancer activity and is
preferred,
whilst against some other cancers, the R-enantiomer exerts more potent anti-
cancer activity
and is preferred, wherein in some embodiments both are independently trialled
against a
cancer (in vivo and/or ex vivo) to see which exerts the greater anti-cancer
activity, wherein
administration is subsequently delimited to the enantiomer, or sample with
enantiomeric
excess for that enantiomer, that is found to have greater anti-cancer activity
against that
particular cancer, and/or the racemate or a scalemate is administered,
optionally with another
compound(s) of this invention, or a pharmaceutically-acceptable salt, solvate,
hydrate or
prodrug thereof.
In some embodiments, the compound is a compound according to the formula
CA 3050553 2019-07-25
R3
Z R2N. /
N
\\CH)N3
.=`
fk S N
N N
I
R1
(R4)q
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein S
symbolises the S stereoisomer, for example, in enantiomeric excess (ee)
exceeding 70%.
In some embodiments, the compound is a compound according to the formula
Z R2''=.. ,R3
N
C*
R'10/N N ilk
H N
I
R1
(R4)q
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein R
symbolises the R stereoisomer, for example, in enantiomeric excess (ee)
exceeding 70%.
In some embodiments, the compound is a compound according to the formula
Z R2NN/R3
N/INN N
H
I
R1
(R4)q. OH
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof.
In some embodiments, the compound is a compound according to the formula
21
CA 3050553 2019-07-25
R2 R3
Z HO iN,
H
I
R1
(R4)q
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein R
symbolises the R stereoisomer, for example, in enantiomeric excess (ee)
exceeding 70%.
In some embodiments, the compound is a compound according to the formula
R3
Z R2 R3
\F
th /2' N.//s, N
HI
(R4),4 R1
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein R
symbolises the R stereoisomer, for example, in enantiomeric excess (ee)
exceeding 70%.
In some embodiments, the compound is
N
N/'=:Th..,..
,---N
N 0
CI H I
NZNNFI
H
CI
11.
CI
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof.
22
CA 3050553 2019-07-25
In some embodiments, the compound is
N
0
S NZNNH
CI
CI
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof. In
some
embodiments the enantiomeric excess (ee) of the S stereoisomer exceeds 70%.
In some embodiments, the compound is
N
411
0
CI D I
N/XNH
CI
101111
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein D is
deuterium (enrichment, for example, exceeding 40% deuterium incorporation at
shown
position, and optionally at other positions also).
In some embodiments, the compound is
23
CA 3050553 2019-07-25
..,....' N
O .../...
N7'',.....7
.
.....--N
N 0
CI \\D A
S N NH
H
CI
411/11)
ci
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein D is
deuterium (enrichment, for example, exceeding 40% deuterium incorporation at
shown
position, and optionally at other positions also). In some embodiments the
enantiomeric
excess (ee) of the S stereoisomer exceeds 70%.
In some embodiments, the compound is
_00-- N
110 ../...
N=/ ..:::., 1,
.....--N
N 0
CI CH3 I
NZXNH
H
CI
41111
CI
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof.
In some embodiments, the compound is
24
CA 3050553 2019-07-25
N
0
CI
NH
CI
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof. In
some
embodiments the enantiomeric excess (ee) of the S stereoisomer exceeds 70%.
In some embodiments, the compound is
N
410
0
CI CH3A
CI
NH
CI
41111
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof. In
some
embodiments the enantiomeric excess (ee) of the R stereoisomer exceeds 70%.
In some embodiments, the compound is
CA 3050553 2019-07-25
OH
0
CI
N/NNH
CI
CI
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof.
In some embodiments, the compound is
N
(010
OH
0
CI
W'oiNZNNH
CI
01111
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof. In
some
embodiments the enantiomeric excess (ee) of the R stereoisomer exceeds 70%.
In some embodiments, the compound is
26
CA 3050553 2019-07-25
1110
0
NH
CI
CI
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof. In
some
embodiments the enantiomeric excess (ee) of the R stereoisomer exceeds 70%.
Compounds of this invention, which reduce FiF0 ATP hydrolysis, can also be
used to
treat/ameliorate/prevent/combat other diseases, disorders and conditions.
Compounds of this
invention attack cancer characteristics shared with embryonic stem cells,
which aren't found
in the adult human body, but are in the blastocyst ¨5 days after
fertilization. Thus compounds
of this invention have utility as emergency contraceptives, for preventing
unwanted
pregnancy, with a later time window than the "morning after pill". Reducing
FIFO ATP
hydrolysis reduces a futile cycle of ATP synthesis and hydrolysis, used by the
body for heat
generation (supporting mouse data herein). If exogenous heat replaces this
reduced
endogenous heat (higher room temperature, wearing more clothes, geographical
relocation to
the tropics etc.), this reduces energy (food) consumption and
.. treats/ameliorates/prevents/combats cachexia, cancer driven cachexia and
weight loss,
wherein cachexia is the biggest cause of death in cancer patients. Reducing
this ATP
synthesis/hydrolysis cycle means the oxidative phosphorylation rate is slower,
less ROS are
produced and the body accumulates less ROS damage per unit time i.e. aging
slows.
Therefore, FIE) ATP hydrolysis inhibitors of this invention extend lifespan
and healthspan,
.. can treat/ameliorate/prevent/combat accelerated aging diseases, progeroid
syndromes and the
diseases of aging (e.g. Alzheimer's disease, dementia, Parkinson's disease,
cancer etc.). It is
noteworthy that compounds of this invention both treat cancer and slow aging,
whereas many
present cancer treatments accelerate aging, causing greater incidence of age
related disease(s)
27
CA 3050553 2019-07-25
and ailments. Also, it is noteworthy that compounds of this invention both
treat and prevent
cancer, whereas many present cancer treatments (e.g. radiotherapy) increase
cancer risk.
Activated macrophages are distinct from resting macrophages, and other normal
adult cells,
because the nitric oxide they produce to kill pathogens switches off their use
of oxidative
phosphorylation and they rely on FIE) ATP hydrolysis to maintain LPN.
Compounds of this
invention inhibit FiF0 ATP hydrolysis and so depolarise TN in activated (not
resting)
macrophages, which triggers their apoptosis. Compounds of this invention
treat/ameliorate/prevent/combat macrophage associated diseases or disorders
(e.g.
Macrophage Activation Syndrome, HIV hides safely in activated macrophages
during anti-
retroviral therapy {ART} and from here repopulates HIV virus in blood plasma
when ART is
interrupted or discontinued, virus neuroinvasion via macrophages, thence HIV-
associated
neurocognitive disorders). FIR ATP hydrolysis inhibitors, by increasing
metabolic/bioenergetic efficiency (less heat produced), can cause
energy/weight gain in a
subject, which has therapeutic, aesthetic, physical/mental performance
applications, and
commercial applications in livestock and farming. Compounds of this invention
reduce FIN
ATP hydrolysis and can reduce body temperature to a value controlled by
intersection of
compound dosage and ambient temperature (even at maximum possible effect,
compound
can't make body fall below, only to, ambient temperature; body temperature
controlled by
controlling ambient temperature), which can treat/ameliorate/prevent/combat a
disease or
disorder that causes a higher than normal body temperature (e.g. fever,
infection, sepsis,
malignant hyperthermia, neuroleptic malignant syndrome etc.) and a disease or
disorder
combated (or surgery or medical treatment helped) by hypothermia (e.g.
neuroprotection/cardioprotection/tissue protection after a stroke or ischemia,
deep
hypothermic circulatory arrest for surgery etc.). Disclosed herein are the
first drugs to
treat/ameliorate/prevent/combat emergency grade hyperthermia, wherein
hyperthermia is an
extremely dangerous aspect to many Emergency Room (ER) admissions e.g. in some
trauma
patients. This is a valuable contribution to the art. Inhibiting FIN ATP
hydrolysis reduces
body temperature, which slows/reduces neural activity, wherein as regards body
temperature,
large reduction confers sedation (experimentally observed in mice in data
herein), with
applications to sleep and surgery etc., and smaller reduction confers anti-
hyperactivity, anti-
anxiety, anti-depression, anti-pain and treatment for premature ejaculation,
epilepsy,
Tourette's syndrome, Attention Deficit Hyperactivity Disorder (ADHD), Post
Traumatic
Stress Disorder (PTSD), homicidal/criminal/suicidal/self-harm
ideation/tendency/thoughts
etc. The intersection between PIP ATP hydrolysis inhibitor drug dose, and
ambient
28
CA 3050553 2019-07-25
temperature, dictates how much body temperature falls and thence depth of the
sedation,
wherein if ambient temperature equals 37 C, the drug can't reduce body
temperature below
this, no matter the dose, and no sedative action can occur. Drug action
against a fundamental
physiological parameter (body temperature), which dictates a further
fundamental
physiological parameter (action potential characteristic(s): firing
threshold/conduction
velocity/firing frequency etc.), yields incredibly broad therapeutic
application. It combats any
pathology/condition characterized by too much/inappropriate/undesired
signals/activity/electrical activity in the nervous system. Juxtaposition of
sedation with anti-
aging action, which a compound of this invention confers, has applications to
space travel,
especially because the sedation can be turned on and off by settings of the
ambient
temperature.
An aspect of the invention is a pharmaceutical composition comprising at least
one
(optionally more than one) compound, as described herein, and a
pharmaceutically-
acceptable carrier or excipient or diluent.
An aspect of the invention is a compound(s) and/or composition(s) as described
herein for
use in a method of treatment of the human or animal body by therapy.
An aspect of the invention is a compound(s) and/or composition(s) as described
herein
administered to a subject topically or systemically or both.
An aspect of the invention is a compound(s), or a pharmaceutically-acceptable
salt, solvate,
hydrate or prodrug thereof, and/or composition(s) as described herein,
optionally a
compound(s), and/or composition(s) that reduces FIE) ATP hydrolysis in a
subject, for use in
treating, ameliorating, preventing, reversing or combating a disease or
disorder, or
unwanted/undesirable physiological process or its consequences or an
unwanted/undesirable
aesthetic, selected from the following list;
Encompassed by this invention is a method of treating, ameliorating,
preventing, reversing or
combating a disease or disorder, or unwanted/undesirable physiological process
or its
consequences or an unwanted/undesirable aesthetic, in a subject, selected from
(i) cancer, any cancer, neoplasia, metastasis, tumor
formation/growth/implantation,
tumorigenesis, solid tumor, blood borne tumor, cancer that is refractory or
resistant to
conventional chemotherapy, drug resistant tumor, multidrug resistant cancer;
29
CA 3050553 2019-07-25
(ii) cancer that metabolizes much of its glucose and/or glutamine to lactate,
for
example a cancer exhibiting the Warburg effect and/or a cancer that can be
discriminated
from surrounding tissue by PET imaging (e.g. '8F-FDG PET);
(iii) cachexia, cancer driven cachexia, cancer fatigue, weight loss, weight
loss for
known or unknown reason, chronic wasting disease, atrophy, brown atrophy,
frailty, frailty
syndrome, aging frailty, geriatric syndrome, age-related cachexia and/or
sarcopenia,
weakness, wasting, anorexia nervosa, bulimia nervosa, eating disorder,
amenorrhea,
underweight, low body mass index (BMI, e.g. <18.5), low body fat percentage,
body
composition change, wasting syndrome, HIV wasting syndrome, malnutrition,
clinical
malnutrition, starvation, kwashiorkor syndrome, marasmus syndrome,
malabsorption,
malabsorption due to parasitic/bacterial infection (e.g. helminthiasis,
Whipple's disease, small
intestine bacterial overgrowth (SIB0), giardiasis etc.), anemia, refeeding
syndrome, appetite
loss, catabolysis, muscle atrophy, asthenia, muscle weakness (myasthenia),
sarcopenia,
osteoporosis, cachexia associated with HIV, AIDS, multiple sclerosis,
rheumatoid arthritis,
familial amyloid polyneuropathy, chronic kidney disease, cystic fibrosis,
multiple sclerosis,
motor neuron disease, Parkinson's disease, dementia, Addison's disease,
mercury poisoning
(acrodynia), chronic pancreatitis, untreated/severe type 1 diabetes mellitus,
hormonal
deficiency, tuberculosis, gastroenteritis, diarrhea, dysentery, any digestive
disease or
disorder, any gastrointestinal disease or disorder including functional
gastrointestinal
disorders, coeliac disease, tropical sprue, irritable bowel syndrome,
inflammatory bowel
disease, Crohn's disease, ulcerative colitis, short bowl syndrome, congestive
heart failure,
constrictive pericarditis, bradycardia, chronic obstructive pulmonary disease
(COPD),
altitude sickness, hyperthyroidism (subclinical hyperthyroidism, Graves'
disease,
multinodular goiter, toxic adenoma, inflammation of the thyroid {thyroiditis},
pituitary
.. adenoma), fatigue, chronic fatigue syndrome or any disease or disorder or
pathology in which
a body tissue(s) is undersupplied or underutilises (vs. its need) an
energetic/chemical
substrate(s), including 02;
(iv) cancer associated fever, which is especially associated with, but not
limited to,
non Hodgkin lymphoma (NHL), Hodgkin lymphoma, acute leukaemia, kidney cancer
(renal
cell cancer), liver cancer (hepatocellular carcinoma), bone cancer, adrenal
gland tumours
such as phaeochromocytomas, tumours in the hypothalamus, solid tumours;
(v) disease or disorder or physiological process or condition that causes a
higher than
normal body temperature such as (without limitation) high environmental
temperature,
ingesting an uncoupler (e.g. 2,4-dinitrophenol), infection, sepsis,
neutropenic sepsis, stroke,
CA 3050553 2019-07-25
fever, pyrexia, hyperpyrexia, hyperthermia, malignant hyperthermia,
neuroleptic malignant
syndrome, serotonin syndrome, toxic serotonin syndrome, thyroid storm,
heatstroke, surgery
related, menopause ("hot flushes"), infection (non-limiting e.g. roseola,
measles, enteroviral
infections, parasitic, viral, fungal, Chlamydial, Rickettsial, bacterial,
mycobacterial, systemic
bacterial, intravascular, HIV associated, nosocomial), pyrogenic infection,
thermoregulatory
disorder(s), connective tissue disease(s), Kawasaki syndrome, drug overdose,
drug or drug
withdrawal induced hyperthermia, alcohol/drug withdrawal, idiosyncratic drug
reaction, fever
of known or unknown or uncertain origin (non-limiting e.g. infectious
disease(s),
inflammation, immunological disease(s), non-infectious inflammatory disease(s)
{non-
limiting eg. systemic rheumatic and autoimmune diseases, vasculitis,
granulomatous diseases,
autoinflammatory syndromes}, tissue destruction, reaction to incompatible
blood product(s),
metabolic disorder(s), inherited metabolic disorder(s), cancer, neoplasm,
endogenous or
exogenous pyrogen(s), injury, head injury);
(vi) disease/disorder/injury/pathology/surgery
treatable/ameliorated/prevented/combated/helped by conferring hypothermia in a
subject for
some medical or other purpose which can include slowing a chemical reaction(s)
rate in a
subject for therapeutic benefit, preventing/minimizing brain and/or tissue
damage, slowing
physiological/pathological processes (reaction rates are temperature
dependent) and so
"buying time" to get the subject to treatment for their emergency (e.g.
trauma/septic shock or
other medical emergency), slowing the progress of sepsis until a sufficient
concentration of a
working antibiotic(s) can be built up in the subject (furthermore hypothermia,
by slowing
sepsis progression, buys time to observe which antibiotic(s) can work,
yielding time to try
alternative further antibiotic option(s) if required), used soon after or just
before clinical/legal
death to preserve the subject's organs/tissues until the subject can be
frozen/cryogenically
frozen or the pathology that caused clinical/legal death (e.g. wound) can be
fixed and the
subject resuscitated, administered to a subject when a first responder (e.g.
ambulance crew)
deems the subject dead or unlikely to survive the journey to a medical
facility (e.g. hospital)
wherein this administration helps to preserve the subject which is helpful if
hospital staff
subsequently assess that they can, or might be able to, save the subject,
stabilizing
surgical/trauma/Emergency Room (ER) patients, deep hypothermic circulatory
arrest for
surgery (DHCA, non-limiting applications of DHCA include repairs of the aortic
arch, repairs
to head and neck great vessels, repair of large cerebral aneurysms, repair of
cerebral
arteriovenous malformations, pulmonary thromboendarterectomy, resection of
tumors that
have invaded the vena cava, brain tumor resection {wherein the anti-cancer
activity of a
31
CA 3050553 2019-07-25
compound(s) of this invention juxtaposes well}), Emergency Preservation and
Resuscitation
(EPR), hypothermia for a surgical purpose, protective hypothermia during
surgery and/or
surgery complication, hypothermia to slow/reduce blood loss, hypothermia for
neuro- and/or
cardio- and/or organ/tissue and/or life protection in a subject that has
trauma/brain
trauma/polytrauma/surgery/stroke/ischemic stroke/hemorrhagic stroke/cardiac
arrest/myocardial infarction/hypoxia/shock (including, without limitation, low
volume,
cardiogenic, obstructive, and distributive shock)/sepsis/septic shock/multiple
organ
dysfunction syndrome/systemic inflammatory response syndrome (SIRS)/organ
failure/cytokine storm/anaphylactic shock/seizure/disseminated intravascular
coagulation/blocked
airway/croup/rhabdomyolysis/[head/facial/spinal/chest/abdominal/ballistic/knife
injury/trauma], or some other medical
emergency/condition/disorder/disease/injury/operation,
hypothermia for cardiac and/or cardiovascular surgery and/or open heart
surgery and/or brain
surgery (neurosurgery) and/or surgery using total circulatory arrest and/or
surgery using
cardiopulmonary bypass, Emergency Preservation and Resuscitation (EPR),
preserving
detached body parts such as limbs and/or organs (for example during organ
storage/transport
and/or transplant, thus increasing the time window for transplantation of
organs to recipients;
compound(s) of this invention is administered to organ to be transplanted [by
administration
to donor and/or by administration to isolated organ] and/or to organ
recipient, optionally
during transplant operation), protective hypothermia, targeted temperature
management,
therapeutic hypothermia, hypothermia therapy for neonatal encephalopathy,
neonatal
hypoxia-ischemia, birth asphyxia, hypoxic-ischemic encephalopathy (HIE),
haemorrhage,
hypovolemia, exsanguination, suspended animation, decompression sickness, burn
injury(s)
including skin bum, inflammation, allergic reaction, anaphylaxis, tissue/organ
rejection,
.. hypoxia, hypoxemia, anoxemia, anoxia, anemia, hypervolemia, altitude
sickness, obstructed
airway, asthma attack, hypoxia in a body/tissue/organ, hypoglycemia,
reperfusion injury
(ischemia-reperfusion injury), upon release of a ligature or tourniquet,
uraemia, crush
syndrome, compartment syndrome, traumatic brain and/or spinal cord injury,
major trauma,
infection, bacterial and/or viral infection(s) (non-limiting e.g. meningitis),
sepsis, septic
shock, stroke, cerebrovascular disease, ischemic brain injury, ischemic
stroke, traumatic
injury, brain injury, spinal cord injury, cardiac arrest, heart failure,
congestive heart failure,
Dilated cardiomyopathy, valvular heart disease, pulmonary embolism, adrenal
crisis,
Addisonian crisis, hypertensive emergency, haemorrhagic (hypovolemic) shock,
cardiogenic
shock, neurogenic shock, hepatic encephalopathy, blood loss, ischemic
32
CA 3050553 2019-07-25
brain/heart/kidney/intestinal injury, neuroprotection and/or cardioprotection
and/or tissue
protection during/after a stroke and/or ischemia and/or cardiac arrest and/or
resuscitation
and/or a period(s) of poor blood flow anywhere in a subject;
(vii) poisoning by a toxic amount of a compound(s) in a subject (non-limiting
e.g.
carbon monoxide/methanol/heavy metal/ethylene glycol/pesticide poisoning,
snake/spider/bee/insect/lizard venom, metabolic poison(s), nerve agent,
chemical weapon,
bacterial toxin(s) (e.g. food poisoning, Salmonella poisoning), endotoxemia,
eukaryote
produced toxin(s) e.g. (non-limiting) brevetoxin, drug(s)/substance(s)
overdose e.g. (non-
limiting) heroin, ethanol, a prescription medication(s), an over the counter
medication(s) such
as aspirin, paracetamol etc.; hypothermia is protective to toxic insult);
(viii) hypermetabolism (optionally because of one or more of, without
restriction,
traumatic brain injury, injury to the body, infection, sepsis, burn, multiple
trauma, fever,
long-bone fracture, hyperthyroidism, prolonged steroid therapy, surgery, bone
marrow
transplant, recovery from anorexia/bulimia), heat intolerence, insomnia, fatal
insomnia,
nervousness, Luft's disease, non-thyroidal hypermetabolism, thyrotoxicosis,
hyperthyroidism,
overactive thyroid, subclinical hyperthyroidism, too much thyroid hormone(s)
in the subject,
too much triiodothyronine (T3) and/or thyroxine (T4) in the subject,
hyperthyroxinemia
(including, without restriction, familial dysalbuminemic hyperthyroxinemia,
familial
euthyroid hyperthyroxinemia, thyroid hormone resistance syndrome), thyroid
storm,
hyperthyroidism caused by one or more of (without restriction) Graves'
disease, thyroiditis,
Hashimoto's thyroiditis, subacute thyroiditis, postpartum thyroiditis, lumps
(nodules) on the
thyroid, enlarged thyroid gland (goitre), simple goitre, multinodular goiter,
toxic multinodular
goiter, toxic adenoma, toxic thyroid adenoma, inflammation of the thyroid,
hyperplasia of
thyroid, metastatic thyroid cancer, thyroid tumour, thyroid cancer (including,
without
restriction, papillary carcinoma, follicular carcinoma, medullary thyroid
carcinoma,
anaplastic thyroid carcinoma), eating too much iodine, goitrogen ingestion,
consumption of
ground beef contaminated with thyroid tissue ("hamburger hyperthyroidism"),
too much
synthetic thyroid hormone in the subject, pituitary adenoma, drug induced,
Amiodarone drug
induced, struma ovarii, Jod-Basedow syndrome, nonautoimmune autosomal dominant
hyperthyroidism;
(ix) low or less than desired metabolic/bioenergetic efficiency in a subject,
or low or
less than desired physical or mental performance (e.g. memory, IQ), or low or
less than
desired body weight, or fatigue/tiredness/weakness/exhaustion;
33
CA 3050553 2019-07-25
(x) accelerated aging disease or progeroid syndrome including, without
restriction,
Werner syndrome, Bloom syndrome, De Barsy syndrome, Rothmund¨Thomson syndrome,
Cockayne syndrome, xeroderma pigmentosum, trichothiodystrophy, combined
xeroderma
pigmentosum-Cockayne syndrome, restrictive dermopathy, Wiedemann¨Rautenstrauch
syndrome, Hutchinson¨Gilford progeria syndrome (progeria), Ataxia
telangiectasia-like
disorder 2, XFE progeroid syndrome, Muscular dystrophy, Muscular Dystrophy
(Becker's,
Duchenne, Limb-Girdle), Yamamoto's Muscular Dystrophy, Mandibuloacral
dysplasia,
Dilated cardiomyopathy, GAPO syndrome, Cutis laxia, Ehlers-Danlos syndrom,
Lenz-
Majewski hyperostatic dwarfism, SHORT syndrome, Progressive external
opthalmoplegia,
.. Nester-Guillermo progeria syndrome, MDPL syndrome, Dyskeratosis congenital,
Down
syndrome;
(xi) disease or disorder of aging (incidence/severity increases with increased
age/senescence) and/or unwanted/undesirable aspect(s) of aging and/or a
disease/disorder
associated with elevated reactive oxygen species including age-associated
decline, aging
frailty, frailty syndrome, sarcopenia, muscle weakness, osteoporosis,
cognitive decline,
cognitive defecit, mild cognitive impairment, degenerative diseases,
neurodegenerative
diseases, motor-associated neurodegenerative diseases, motor neuron disease,
amyotrophic
lateral sclerosis (ALS), primary lateral sclerosis, progressive muscular
atrophy, progressive
bulbar palsy, progressive supranuclear palsy, pseudobulbar palsy, hereditary
spastic
paraplegia, Parkinson's disease, Multiple System Atrophy (MSA), Progressive
Supranuclear
Palsy (PSP), essential tremor, resting tremor, Alzheimer's disease,
Huntington's disease,
spinocerebellar ataxias, Friedreich's ataxia, dementia, frontotemporal
dementia, chronic
traumatic encephalopathy, memory loss, aged cognition, age/aging related
cognitive
decline/impairment, Batten disease, polyglutamine diseases, osteoporosis,
atherosclerosis,
cardiovascular disease, myocardial infarction, cerebrovascular disease,
stroke, heart failure,
heart failure with preserved ejection fraction, idiopathic pulmonary fibrosis,
fibrotic disease,
pulmonary disease, coronary artery disease, hypercholesterolemia, obesity,
liver disease, fatty
liver disease, lysosomal storage disease, amyloidosis, systemic sclerosis,
kidney disease,
hepatic cirrhosis, immunosenscence, clonal hematopoiesis, chronic obstructive
pulmonary
disease (COPD), hypertension, hypercholesterolemia, age-related thymic
atrophy, arthritis,
osteoarthritis, arthritis (Osteo-and Rheumatoid), Juvenile Rheumatoid
Arthritis (JRA),
Degenerative Disc Disease, Tendinopathy, Androgenetic Alopecia, male-pattern
baldness,
Idiopathic Pulmonary Fibrosis, systemic sclerosis, Chronic Obstructive
Pulmonary Disease,
Psoriasis, age-related loss of cardiac/pulmonary/cognitive/vision function,
diabetes, type 2
34
CA 3050553 2019-07-25
diabetes, andropause, glaucoma, retinal degeneration, sarcopenia, cachexia,
age-related
cachexia and/or sarcopenia, age-related macular degeneration (AMD,
early/intermediate/late), neovascular/wet AMD, dry AMD, Geographic atrophy
(GA), wet
and dry AMD in the same eye(s), Stargardt's macular degeneration, Best
vitelliform macular
dystrophy, diabetic retinopathy, proliferative diabetic retinopathy, diabetic
macular edema,
age/aging-related eye disease, ophthalmological disease/disorder, ocular
disease, vision loss,
progressive vision impairment, myopia (short-sightedness), degenerative
myopia, hyperopia
(far-sightedness), accommodative dysfunction, glaucoma, cataract formation,
retinal
degeneration, progressive retinal degeneration, retinitis pigmentosa, leber
hereditary optic
neuropathy, Fuchs spot, Best's disease, Sorsby's fundus dystrophy, hearing
loss (e.g. age-
related), presbycusis, tinnitus, naive T cell shortage, movement disability,
nonalcoholic fatty
liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), immunosenescence,
respiratory/urinary tract infection (RTI/UTI) especially in older/aged/elderly
subjects, cancer;
(xii) aging and/or one or more signs of aging, wherein one or more of these
compounds slow/delay/reduce/treat/prevent/stop/reverse aging, and/or extend
lifespan and/or
healthspan, and/or treat or delay the onset of geriatric aging of the
human/animal body,
tissue(s), or organ(s), and/or treat or delay the onset of an age-associated
phenotype in a
cell(s)/organism(s), and/or prolong fertility e.g. female fertility, delay
menopause;
(xiii) skin aging and/or damage (including sun damage) and/or scalp and/or
hair aging
and/or hair greying and/or hair loss;
(xiv) insomnia, fatal insomnia, sleep onset latency, delayed sleep phase
disorder,
exploding head syndrome, parasomnia, sleep-maintenance insomnia, sleep
disorder, too
much/inappropriate/undesired signals/activity/electrical activity in the
nervous system,
hyperactivity, hypersensitivity, Premature ejaculation, hyperreflexia,
Autonomic dysreflexia
(AD), Hyperventilation syndrome, brain hyperactivity, overly sensitive sensory
system,
pathological crying and/or laughing, Pseudobulbar affect (PBA, emotional
lability),
photophobia, phonophobia, temperature-sensitive, pressure-sensitive, brain
hyperexcitability,
overstimulation, intrusive thought(s), Perseveration, sensory overload,
disorganized thinking,
fantasy prone personality, malapdative daydreaming, dissociation, hyperkinetic
disorder,
.. agitation, Psychomotor agitation, restlessness, difficulty controlling
behaviour, disruptive
behaviour disorder, Emotional and behavioral disorder, pervasive developmental
disorder,
Overactive disorder associated with mental retardation and stereotyped
movements, attention-
deficit disorder, Attention Deficit Hyperactivity Disorder (ADHD), adult
attention-deficit
hyperactivity disorder, severe behavioral problem(s) in children (e.g., to
illustrate and not
CA 3050553 2019-07-25
restrict, combativeness and/or explosive hyperexcitable behavior {out of
proportion to
immediate provocation[s]}, hyperactive children who show excessive motor
activity with
accompanying conduct disorders consisting of one or more of: impulsivity,
difficulty
sustaining attention, aggressivity, mood lability, poor frustration
tolerance), Premenstrual
.. dysphoric disorder (PMDD), premenstrual syndrome (PMS), impulsiveness,
impulsivity,
impulse control disorder, lack of self-control, hysteria, histrionic
personality disorder,
attention difficulty, inattention, poor attention control, anxiety, paranoid
anxiety, Paranoid
personality disorder, distress, dysphoria, Adjustment disorder, separation
anxiety, anxiety
disorder, depressive anxiety, agitated depression, treatment-resistant
depression, Generalized
anxiety disorder, social anxiety disorder, stranger anxiety, separation
anxiety (e.g. in dogs left
at home), separation anxiety disorder, Mixed anxiety-depressive disorder,
depression (all
forms, all severities), restlessness/apprehension/anxiety before surgery.
hypochondria, panic
disorder, panic attack, emotional outburst, emotional instability,
Intermittent explosive
disorder, unreasonable/unwarranted anger/aggression, hyper-aggression,
hostility, rage, poor
temper control, self-hatred, poor attentional control, worry, irritability,
neuroses, somatization
disorder, somatic symptom disorder, pain disorder, psychological pain,
psychogenic pain,
psychogenic facial pain, Atypical odontalgia (AO), burning mouth syndrome,
throbbing,
toothache/pulpitis/dental pain, chronic lower back pain, negative emotion,
persistent/enduring
negative emotion, body dysmorphic disorder, factitious disorder, illness
anxiety disorder,
unwarrented fight-or-flight response, stress, emotional stress, emotional
dysregulation,
distress, psychological stress, acute stress, chronic stress, acute stress
reaction, combat stress
reaction, traumatic grief, grief, grief after death of loved one, Prolonged
grief disorder (PGD),
heartbreak, guilt, shame, remorse, emotional pain, Algopsychalia, psychalgia,
suffering,
emotional trauma, psychological trauma, broken heart, Post Traumatic Stress
Disorder
(PTSD), Complex post-traumatic stress disorder (C-PTSD), hypervigilance,
sympathetic
hyperactivity, inability or impaired ability to relax, flashbacks, dysphoric
hyperarousal,
agoraphobia, insomnia, fatal insomnia, mood disorder, irrational/unwarrented
fear/terror,
phobia, social phobia, Cancerophobia, thunderstorm/firework phobia,
hypersexuality,
hypersexual disorder, depression, clinical depression, unipolar depression,
bipolar disorder,
Bipolar I, Bipolar II, Bipolar disorder not otherwise specified (Bipolar NOS),
cyclothymia,
cyclothymic disorder, mixed affective state, atypical depression, melancholic
depression,
postpartum depression, double depression, seasonal affective disorder, mania,
manic episode,
hypomania, increase in energy of psychomotor activity, delirium, excited
delirium, major
depressive disorder, minor depressive disorder, recurrent brief depression,
Depressive
36
CA 3050553 2019-07-25
Disorder Not Otherwise Specified (DD-NOS), major depressive episode,
persistent
depressive disorder (PDD), dysthymia, dysthymic disorder, absence of euthymia,
manic
thoughts, racing thoughts, thought disorder, disordered thinking, reduced
ability to plan and
execute tasks, paranoia, hallucination (including, without limitation, visual,
auditory,
olfactory, gustatory, tactile, proprioceptive, equilibrioceptive, nociceptive,
thermoceptive,
chronoceptive), Charles Bonnet syndrome, delusion, persecutory delusion,
hearing voices,
homicidal/criminal ideation/tendency/thoughts, suicidal
ideation/tendancy/thoughts, self-
injury, non-suicidal self-injury, violence, attacking others, negative mood
swing, personality
disorder, Borderline personality disorder, Narcissistic personality disorder,
malignant
narcissism, dissociative disorder, dissociative identity disorder (DID),
Psychosis, acute
psychosis, chronic psychosis, psychosis spectrum disorder, manifestations of
psychotic
disorders, behavioral complications of mental retardation, stimulant
psychosis, psychotic
depression, hallucinogen persisting perception disorder, Psychoactive
substance-related
disorder, amphetamine-induced psychosis, brief psychotic disorder, Brief
reactive psychosis,
Menstrual psychosis, postpartum psychosis, Psychotic disorder, Psychopathy,
chronic
hallucinatory psychosis, manifestation(s) of psychotic disorder,
neurotic/reactive/endogenous/involutional/psychotic depression/depressive
disorder
(optionally accompanied by anxiety or agitation), depressive neurosis,
delusional depression,
psychotic aggression, psychiatric symptoms of dementia, AIDS delirium,
Supersensitivity
psychosis, Tardive psychosis, Tardive dysmentia, Depersonalization disorder,
out-of-body
experience, Sociopathy, Schizophrenia, Paranoid schizophrenia, disorganized-
type
schizophrenia, simple-type schizophrenia, pseudoneurotic schizophrenia,
prodromal
schizophrenia, schizoaffective disorder, bipolar type schizoaffective
disorder, depressive type
schizoaffective disorder, schizoaffective psychosis, Schizotypal personality
disorder,
schizophreniform disorder, Delusional parasitosis, formication, paresthesias,
Acroparesthesia,
tinnitus, delusional disorder, delusional jealousy, inappropriate behaviour,
behavioural
disorder, antisocial personality disorder, Oppositional defiant disorder
(ODD), conduct
disorder (CD), Disruptive mood dysregulation disorder (DMDD), sadistic
personality
disorder, poor inhibitory control, kleptomania, Pyromania, trichotillomania,
dermatillomania,
pathological/problem gambling, dyskinesia, tardive dyskinesia, paroxysmal
dyskinesia,
Paroxysmal kinesigenic dyskinesia, Paroxysmal nonkinesigenic dyskinesia,
Paroxysmal
exercise-induced dystonia, Hemiballismus, tic disorder, tremor, clonus,
Tourette's syndrome,
coprolalia, copropraxia, Echophenomena, Echopraxia, Echolalia, Palilalia,
stuttering/stammering, stereotypy, punding, Self-stimulatory behaviour
(stimming),
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CA 3050553 2019-07-25
Stereotypic movement disorder (SMD), synesthesia, obsession,
Obsessive¨compulsive
disorder (OCD), obsessive¨compulsive personality disorder, anankastic
personality disorder,
relationship obsessive¨compulsive disorder (ROCD), Scrupulosity, Primarily
obsessional
obsessive compulsive disorder, sexual obsession, Akathisia (including, without
limitation,
chronic, acute, Pseudoakathisia, Tardive akathisia, withdrawal or "rebound"
akathisia),
Restless legs syndrome, motor restlessness, periodic limb movement disorder
(PLMD),
periodic limb movements in sleep (PLMS), Sydenham's chorea, chorea, dystonia,
Hypnic
jerk, twitching, spasms, Tetanus, tetanus muscle spasms, tetanized state,
Myoclonus,
myoclonic seizure, Action myoclonus, Palatal myoclonus, Middle ear myoclonus,
Spinal
myoclonus, Stimulus-sensitive myoclonus, Sleep myoclonus, Cortical reflex
myoclonus,
Essential myoclonus, myoclonic epilepsy, Juvenile myoclonic epilepsy,
Progressive
myoclonus epilepsy (PME, including, without limitation, Dentatorubral-
pallidoluysian
atrophy, Unverricht-Lundborg disease, MERRF syndrome, Lafora disease),
Reticular reflex
myoclonus, Myoclonic epilepsy, diaphragmatic flutter, automatism, status
epilepticus,
Epilepsia partialis continua, Complex partial status epilepticus, epilepsy,
epileptic seizure,
simple partial seizure, complex partial seizure, generalized epilepsy,
generalized seizure
(including, without limitation, tonic-clonic, tonic, clonic, myoclonic,
absence (including
typical absence and atypical absence), atonic seizure), focal epilepsy, focal
seizure,
focal/partial seizure (including, without limitation, Simple partial seizure
and Complex partial
seizure), focal aware seizure, focal impaired awareness seizure, generalised
epilepsy,
temporal lobe epilepsy (including, without restriction, mesial temporal lobe
epilepsy
{MTLE} and lateral temporal lobe epilepsy {LTLE}), Frontal lobe epilepsy,
Rolandic
epilepsy, Nocturnal epilepsy, Nocturnal frontal lobe epilepsy, Autosomal
dominant nocturnal
frontal lobe epilepsy (ADNFLE), Generalized epilepsy with febrile seizures
plus (GEFS+),
.. Panayiotopoulos syndrome, epileptic fit, reflex epilepsy, reflex seizure,
absence seizure
(including, without limitation, childhood absence epilepsy, epilepsy with
myoclonic
absences, juvenile absence epilepsy, juvenile myoclonic epilepsy, Jeavons
syndrome {eyelid
myoclonia with absences}, genetic generalised epilepsy with phantom absences),
complex
partial seizure, atonic seizure, generalized tonic-clonic seizure,
tonic¨clonic seizure, extrinsic
stimulus epilepsy, intrinsic stimulus epilepsy, photosensitive epilepsy,
musicogenic epilepsy,
thinking epilepsy, eating epilepsy, seizure(s), Febrile seizure, nerve agent
induced seizure,
Dravet syndrome (sometimes modest hyperthermic stressors like physical
exertion or a hot
bath can provoke seizures in affected individuals), acute symptomatic seizure,
seizure-related
disorder, drug related seizure, paroxysmal depolarizing shift, Ohtahara
syndrome, Epilepsy in
38
CA 3050553 2019-07-25
females with mental retardation, Rasmussen's encephalitis, Epilepsy syndrome,
benign
rolandic epilepsy, childhood absence epilepsy, absence epilepsy, juvenile
myoclonic
epilepsy, epileptic encephalopathies, Lennox¨Gastaut syndrome, West syndrome
(Epileptic
spasms), Doose syndrome (Myoclonic astatic epilepsy, MAE), Lennox-Gestaut
syndrome,
pseudo-Lennox Gastaut syndrome (atypical benign partial epilepsy), Benign
familial neonatal
epilepsy, Benign occipital epilepsy of childhood, familial neonatal
convulsions, Febrile
infection-related epilepsy syndrome, interictal dysphoric disorder, euphoria
sclerotic,
psychogenic non-epileptic seizure, non-epileptic seizure, gelastic seizure,
convulsion(s),
migraine, status migrainosus, tension headache, headache, Hypnic headache,
hiccups,
intractable hiccups, thumps in equines, Postural orthostatic tachycardia
syndrome (POTS),
Pheochromocytoma, agony, pain, chronic pain, acute pain, pain due to
disease/injury,
neuropathic pain, fibromyalgia, postherpetic neuralgia, phantom pain, referred
pain, back
pain, lower back pain, pelvic pain, cancer associated pain, chemotherapy
associated pain,
Diabetic neuropathy, small fiber peripheral neuropathy, Mononeuritis
multiplex,
Wartenberg's migratory sensory neuropathy, Breakthrough pain, idiopathic pain,
polyneuropathy, Neuritis, Mononeuropathy, Polyradiculoneuropathy, Radial
neuropathy,
neuropathy, visceral pain, Burning feet syndrome, Tarsal tunnel syndrome,
Carpal tunnel
syndrome, Guyon's canal syndrome, Cubital Tunnel Syndrome, Repetitive strain
injury,
Sciatica, Neurofibromatosis, Ulnar nerve entrapment, Erythromelalgia,
Paroxysmal extreme
pain disorder, Proctalgia fugax, dysesthesia, scalp dysesthesia, dysesthetic
burning,
hyperesthesia, hyperalgesia, Opioid-induced hyperalgesia, hyperpathia,
allodynia, pain
response from stimuli which do not normally provoke pain, Complex regional
pain syndrome
(said to be most painful condition known to man), Radiculopathy, neuralgia
(including,
without restriction, intercostal neuralgia, trigeminal neuralgia, atypical
trigeminal neuralgia,
glossopharyngeal neuralgia, occipital neuralgia, postherpetic neuralgia),
ciguatera poisoning,
irritable bowel syndrome (IBS), interstitial cystitis (IC), temporomandibular
joint disorder,
acute intermittent porphyria, Porphyria, Acute porphyria (including, without
limitation, acute
intermittent porphyria {AIP}, variegate porphyria {VP}, aminolevulinic acid
dehydratase
deficiency porphyria {ALAD}, hereditary coproporphyria {HCP}, drug induced),
Chronic
porphyria (including, without limitation, X-linked dominant protoporphyria
{XLDPP},
congenital erythropoietic porphyria {CEP}, porphyria cutanea tarda {PCT}, and
erythropoietic protoporphyria {EPP}), cutaneous porphyria, Porphyria cutanea
tarda, allergy,
allergic reaction, anaphylaxis, anaphylactic shock, hives, asthma, allergic
rhinitis, rhinitis,
urticaria, contact dermatitis, cough, sore throat, esophageal reflux disease,
heartburn, chest
39
CA 3050553 2019-07-25
pain, Esophageal motility disorder, Nutcracker esophagus, diseases involving
gastrointestinal
motility, Peptic ulcer disease, esophageal spasm, angina, itchiness, Pruritus,
severe pruritus,
Prurigo, Pruritic skin condition, chronic itch, eczema, pruritus in eczema,
neuropathic itch,
neurogenic itch, itchiness due to atopic dermatitis and/or lichen simplex
chronicus, peripheral
sensitization, central sensitization, sensory perception of absent stimuli,
too much sensory
stimulation, sensory stimulation brings discomfort, Neuromyotonia, Peripheral
nerve
hyperexcitability, Morvan's syndrome, Benign fasciculation syndrome, Cramp
fasciculation
syndrome, hyperhidrosis, undifferentiated somatoform disorder, somatoform
disorder,
somatic symptom disorder, conversion disorder, functional neurological symptom
disorder,
severe nausea and/or vomiting, severe nausea/emesis, Chemotherapy-induced
peripheral
neuropathy, chemotherapy-induced nausea and vomiting (CINV), radiation therapy-
induced
nausea and vomiting (RINV), postnarcotic nausea, hyperemesis gravidarum,
morning
sickness, Cyclic vomiting syndrome, retching/dry heaving, Urinary
incontinence, enuresis,
nocturnal enuresis, tail chasing, reduce urine spraying/marking behavior,
benzodiazepine
withdrawal syndrome, barbituate withdrawal syndrome, Neuroleptic
discontinuation
syndrome, drug withdrawal discomfort/pain/symptoms, drug use disorder, alcohol
use
disorder, opoid use disorder, amphetamine use disorder, cocaine use disorder,
alcohol
withdrawal syndrome/symptoms, delirium tremens, opoid withdrawal
sydrome/symptoms,
drug craving, drug addiction, drug dependence, polysubstance dependence, drug
overdose,
smoking, tobacco (nicotine) addition, tobacco (nicotine) withdrawal symptoms,
alcoholism,
addiction, opoid addiction, cocaine/crack addiction, addictive behaviour,
addictive
personality, behavioural addiction, internet/computer/computer game/social
media/media
addiction, exercise addiction, compulsive behaviour (e.g. {non-limiting}
checking, counting,
washing, repeating), anti-social behaviour, criminality, sexual compulsion,
impulsive sexual
behaviour, compulsive buying, gambling addiction, sex related addiction,
sexual urge,
hunger, eating desire/compulsion, eating disorder, polyphagia, overeating,
binge eating
disorder, compulsive overeating, insatiable/excessive appetite, bulimia
nervosa, anorexia
nervosa, substance abuse, substance-induced delirium, substance-induced
psychosis,
substance-induced mood disorder, drug overdose, vertigo, motion sickness,
seasickness,
mental/nervous breakdown, Autism spectrum disorder, neurological disorder,
cognitive
disorder, mental disorder, mental health disorder, mental health condition
involving impaired
or altered neural plasticity, mood disorder, mental disorder disclosed in
Diagnostic and
Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) or a later
edition, a
CA 3050553 2019-07-25
mental/behavioural disorder disclosed by the International Classification of
Diseases (ICD) in
LCD-10 Chapter V: Mental and behavioural disorders (World Health Organisation,
WHO); or
(xv) diseases or disorders or conditions or pathologies or
unwanted/undesirable
effects/actions/behaviour treatable/ameliorated/prevented/combated, in
totality or in part (e.g.
along with surgery), by anaesthesia, pre-anaesthesia, post-anaesthesia,
hypoesthesia,
hypoactivity, sedation, coma, tranquilization, behavioural submission, muscle
relaxation,
hibernation, artificial hibernation, torpor, synthetic torpor, suspended
animation (e.g. used
during spaceflight because e.g. reduces ionizing radiation damage to subject);
(xvi) hyperproliferative/hyperplasia disorder, non-cancerous proliferative
disorder,
hyperproliferative autoimmune disorder, hyperplasia, epidermal hyperplasia,
dysplasia (e.g.
epithelial dysplasia), nodule(s), wart(s), papilloma(s), squamous cell
papilloma, genital
wart(s), condyloma(s), condyloma acuminatum, cyst(s), polyp(s) {including,
without
restriction, digestive, colorectal, endometrial, cervical, nasal, laryngeal,
inflammatory fibroid
polyp[s])}, inherited/hereditary (including, without restriction, Familial
adenomatous
polyposis, Peutz¨Jeghers syndrome, Turcot syndrome, Juvenile polyposis
syndrome, Cowden
disease, Bannayan¨Riley¨Ruvalcaba syndrome {Bannayan-Zonana syndrome},
Gardner's
syndrome) and non-inherited (non-restrictive e.g. Cronkhite¨Canada syndrome)
polyposis
syndrome, benign tumour, adenoma, organ enlargement by hyperplasia, Cushing's
disease
(enlarged adrenal cortex by hyperplasia), congenital adrenal hyperplasia,
hyperplasia of
breast, atypical ductal hyperplasia, intraductal papillomatosis,
fibroadenomas, fibrocystic
changes, hemihyperplasia, focal epithelial hyperplasia, sebaceous hyperplasia,
sebaceous
adenoma, intimal hyperplasia, unwanted/undesirable smooth muscle cell
proliferation,
smooth muscle cell hyperplasia, intimal smooth muscle cell hyperplasia,
neointimal
hyperplasia, proliferative vascular disorders, stenosis, stenosis because of
cellular
proliferation, vaginal stenosis, stenosis in a blood vessel, lessoned patency
of a blood vessel,
stenosis in a blood vessel because of cellular proliferation, vascular
occlusion, restenosis,
restenosis in a blood vessel that has been implanted with a stent, in-stent
restenosis, post-
angioplasty restenosis, systemic sclerosis, cirrhosis of the liver, adult
respiratory distress
syndrome, idiopathic cardiomyopathy, lupus erythematosus, retinopathy (e.g.
diabetic
retinopathy and/or other retinopathy[y/ies]), cardiac hyperplasia, fibrosis,
pulmonary fibrosis,
idiopathic pulmonary fibrosis, fibromatosis, neurofibromatosis, renal
interstitial fibrosis,
Cowden syndrome, hamartoma(s), choristoma(s), hemangioma(s), lymphangioma(s),
rhabdomyoma(s), lymphangiomatosis, cystic hygroma, trichilemmoma, sarcoidosis,
neurosarcoidosis, aggressive fibromatosis, desmoid tumour(s),
unwanted/undesirable skin
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CA 3050553 2019-07-25
cell proliferation, hyperproliferative skin disorder, psoriasis (including,
without restriction,
plaque, guttate, inverse, pustular, napkin, seborrheic-like, nail, scalp and
erythrodermic
psoriasis), psoriatic arthritis, dactylitis, seborrhoeic dermatitis, dandruff,
eczema, atopic
dermatitis, rosacea, reactive arthritis (Reiter's syndrome), pityriasis rubra
pilaris,
hyperproliferative variants of disorders of keratinization (e.g., without
restriction, actinic
keratosis, senile keratosis, keratosis pilaris, seborrheic keratosis),
scleroderma, benign
prostatic hyperplasia, prostate enlargement, endometrial hyperplasia, atypical
endometrial
hyperplasia, benign endometrial hyperplasia, adenomyosis, atypical polypoid
adenomyoma,
endometriosis, endometriosis of ovary (endometrioma), endometrial polyp(s),
polycystic
ovary syndrome, ovarian cyst(s), cervical polyp(s), uterine fibroid(s),
uterine hyperplasia;
(xvii) Tumour Associated Macrophages (TAMs) or any macrophage associated
disease or disorder such as, without limitation, Macrophage Activation
Syndrome (MAS),
HIV, AIDS, HIV-associated neurocognitive disorders (HAND), AIDS dementia, HIV
peripheral neuropathy, HIV associated cancers, AIDS-defining cancers, non-AIDS
defining
cancers, any disease in which the pathogen(s) hides from the immune system in
macrophages
including, without limitation, Mycobacterium tuberculosis (causes
tuberculosis), Leishmania
parasite (causes Leishmaniasis), Chikungunya virus (causes Chikungunya),
Legionella
pneumophila (causes Legionnaires' disease), adenoviruses, T. whipplei (causes
Whipple's
Disease), Brucella spp. (causes brucellosis), Staphylococcus aureus, Ebola
virus, Hepatitis B
virus, Hepatitis C virus, influenza virus strains, dengue virus and antibiotic
resistant bacteria,
any disease or condition in which activated macrophages are unwanted or
undesirable;
(xviii) virus/pathogen neuroinvasion via macrophage(s), as used for non-
limiting
example by HIV, Heptatitis C virus and SARS coronavirus;
(xix) neurocognitive or neurodegenerative diseases/disorders, for non-limiting
example those caused by a virus;
(xx) virus/pathogen transmission from mother to fetus/baby via macrophage(s)
as
used for non-limiting example by zika (via Hofbauer cells) and HIV
(macrophages in breast
milk);
(xxi) acute or chronic or systemic inflammation or any inflammatory
disease/disorder/syndrome or any autoinflammatory disease/disorder/syndrome or
any
autoimmune disease/disorder/syndrome;
(xxii) acute inflammation, chronic inflammation, systemic inflammation,
inflammation because of infection or foreign bodies or injury or chemical or
toxin or drug or
stress or frostbite or burn or ionising radiation or surgery, inflammatory
42
CA 3050553 2019-07-25
diseases/disorders/syndromes, Macrophage Activation Syndrome (MAS),
autoinflammatory
diseases/disorders/syndromes, age-related chronic inflammatory diseases
("inflammaging"),
autoimmune diseases/disorders/syndromes, diseases/disorders of the innate
immune system,
sore throat, sore throat associated with cold or flu or fever, high-intensity
exercise associated
inflammation, ulcerative colitis, inflammatory bowel disease (IBD), irritable
bowel syndrome
(IBS), rheumatoid arthritis, osteoarthritis, inflammatory osteoarthritis,
psoriatic arthritis,
atopic dermatitis, allergic airway inflammation, asthma, inflammation
associated depression,
neuroinflammation, neuropathic pain, exercise-induced acute inflammation,
atherosclerosis,
allergy, hay fever, anaphylaxis, inflammatory myopathies, drug-induced
inflammation,
systemic inflammatory response syndrome, sepsis-related multiple organ
dysfunction/multiple organ failure, microbial infection, acute
brain/lung/hepatic/renal
injuries, lung inflammation, acute lung injury (ARDS), acne vulgaris, celiac
disease, celiac
sprue, chronic prostatitis, colitis, autoimmune hemolytic anemia,
diverticulitis,
glomerulonephritis, proliferative glomerulonephritis, membranous nephropathy,
minimal
change nephrotic syndrome, hidradenitis suppurativa, hypersensitivities,
interstitial cystitis,
Mast Cell Activation Syndrome, mastocytosis, otitis, pelvic inflammatory
disease (PID),
endometritis, reperfusion injury, rheumatic fever, rhinitis, sarcoidosis,
transplant rejection,
parasitosis, eosinophilia, type III hypersensitivity, ischaemia, chronic
peptic ulcer,
tuberculosis, Crohn's disease, hepatitis, chronic active hepatitis, immune
hepatitis, alcoholic
hepatitis, chronic viral hepatitis, ankylosing spondylitis, diverticulitis,
fibromyalgia, systemic
lupus erythematous (SLE), Alzheimer's disease, Parkinson's disease,
neurodegenerative
disease, cardiovascular disease, chronic obstructive pulmonary disease,
bronchitis, acute
bronchitis, bronchiectasis, bronchopneumonia, obliterative bronchiolitis,
appendicitis, acute
appendicitis, bursitis, cystitis, dermatitis, encephalitis, HIV encephalitis,
gingivitis,
meningitis, infective meningitis, myelitis, nephritis, neuritis,
periodontitis, chronic
periodontitis, phlebitis, prostatitis, RSD/CRPS, rhinitis, sinusitis, chronic
sinusitis, tendonitis,
testiculitis, tonsillitis, urethritis, vasculitis, respiratory
bronchiolitis¨associated interstitial
lung disease and desquamative interstitial pneumonia, pneumonia, interstitial
lung disease,
Lofgren syndrome, Heerfordt syndrome, monocytosis, liver fibrosis,
steatohepatitis,
.. nonalcoholic steatohepatitis, silicosis, histiocytoses, Langerhans' cell
histiocytosis,
haemophagocytic lymphohistiocytosis, pulmonary langerhans cell histiocytosis,
obesity, type
II diabetes, gout, pseudogout, organ transplant rejection, epidermal
hyperplasia, chronic
fatigue syndrome, graft versus host disease (GVHD), lymphadenopathy,
rheumatoid arthritis
(RA), osteoarthritis (OA), inflammatory osteoarthritis, lupus, multiple
sclerosis (MS),
43
CA 3050553 2019-07-25
myocarditis, uveitis, CNS disease(s), inflammation aspect to a CNS disease(s),
hypothalamic
inflammation, dementia, glaucoma, amyloid related/driven disease, lipid
storage disease(s),
fibrosis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic
steatohepatitis (NASH),
cirrhosis, cirrhosis of the liver, alcoholic cirrhosis, nephropath(y/ies),
lupus nephritis,
immune nephritis, fibrotic disorder(s), cardiovascular disease, heart disease,
atherosclerosis,
vulnerable plaque, plaque formation, lipid containing macrophage related
disease(s)/disorder(s)/malad(y/ies), macrophage foam cells, diabetes, type 1
diabetes, type 2
diabetes, insulin resistance, macrophage aspect to insulin resistance,
obesity, obesity
associated inflammation, macrophage accumulation/large numbers of macrophages
in
adipose tissue (e.g. associated with obesity), granuloma(s), granulomatous
diseases,
sarcoidosis (including, without limitation, Annular sarcoidosis, Erythrodermic
sarcoidosis,
Ichthyosiform sarcoidosis, Hypopigmented sarcoidosis, Lofgren syndrome, Lupus
pernio,
Morpheaform sarcoidosis, Mucosal sarcoidosis, Neurosarcoidosis, Papular
sarcoid, Scar
sarcoid, Subcutaneous sarcoidosis, Systemic sarcoidosis, Ulcerative
sarcoidos),
neurosarcoidosis, pulmonary sarcoidosis, interstitial lung disease, pulmonary
fibrosis,
pulmonary tuberculosis, immune reconstitution syndrome of HIV,
Jarisch¨Herxheimer
reaction, sepsis, Paget's disease of bone, osteolysis, monocytosis,
histiocytosis, X-type
histiocytoses, non-X histiocytoses, Langerhans cell histiocytosis, non-
Langerhans-cell
histiocytosis, malignant histiocytosis, malignant histiocytic disorders,
histiocytomas,
histiocytic lymphoma, hemophagocytic syndrome, hemophagocytic
lymphohistiocytosis,
lymphohistiocytosis, diffuse histiocytic sarcoma, Rosai¨Dorfman disease,
gliosis, Bergmann
gliosis, Chronic Obstructive Pulmonary Disease (COPD), chronic inflammatory
lung disease,
familial mediterranean fever (FMF), TNF receptor-associated periodic syndrome
(TRAPS),
Hyperimmunoglobulinemia D with recurrent fever syndrome (HIDS), cryopyrin
associated
periodic syndrome (CAPS), Blau syndrome, Majeed syndrome, deficiency of
interleukin-1
receptor antagonist (DIRA), mevalonate kinase deficiency, pyogenic-arthritis-
pyoderma
gangrenosum and acne syndrome (PAPA), periodic fever aphthous stomatitis
pharyngitis
adenitis (PFAPA) syndrome, Behcet's disease, Still's disease, Crohn's disease,
Schnitzler's
syndrome, Sweet's syndrome, NLRP12-associated autoinflammatory disorders,
deficiency of
interleukin-1 receptor antagonist (DIRA), pyoderma gangrenosum, cystic acne,
aseptic
arthritis, periodic Fever Associated with mevalonate kinase deficiency
(hyperimmunoglobulin D Syndrome), Pyogenic Arthritis Pyoderma Gangrenosum Acne
(PAPA) syndrome, Periodic Fever Aphthous Stomatitis, Pharyngitis and
Adenopathy
(PFAPA) syndrome, Adult-Onset Still's Disease (AOSD), Systemic Juvenile
Idiopathic
44
CA 3050553 2019-07-25
Arthritis (sJIA), Chronic Recurrent Multifocal Osteomyelitis (CRMO), Synovitis
Acne
Pustulosis Hyperostosis Osteitis (SAPHO) syndrome, Cryopyrin associated
Periodic
Syndrome (CAPS), Familial cold auto inflammatory syndrome (FCAS), Muckle-Wells
syndrome (MWS), Familial cold urticarial, Neonatal onset multisystemic
inflammatory
disorder (NOMID), hereditary Periodic Fever Syndromes, Periodic Fever
Syndromes,
systemic autoinflammatory diseases, Addison's disease, Agammaglobulinemia,
Alopecia
areata, Amyloidosis, Ankylosing spondylitis, Anti-GBM/Anti-TBM nephritis,
Antiphospholipid syndrome, autoimmune angioedema, autoimmune dysautonomia,
autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune inner ear
disease
(AIED), autoimmune myocarditis, autoimmune pancreatitis, autoimmune
retinopathy,
autoimmune urticaria, axonal & neuronal neuropathy (AMAN), Bak) disease,
Behcet's
disease, benign mucosal pemphigoid, Bullous pemphigoid, Castleman disease
(CD), Celiac
disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy
(CIDP),
chronic recurrent multifocal osteomyelitis (CRMO), Churg-Strauss, Cicatricial
pemphigoid,
Cogan's syndrome, cold agglutinin disease, congenital heart block, coxsackie
myocarditis,
CREST syndrome, Berger's disease, dermatitis herpetiformis., dermatomyositis,
Devic's
disease (neuromyelitis optica), discoid lupus, Dressler's syndrome,
endometriosis,
eosinophilic esophagitis (EoE), eosinophilic fasciitis, erythema nodosum,
essential mixed
cryoglobulinemia, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant
cell arteritis
(temporal arteritis), giant cell myocarditis, glomerulonephritis,
proliferative
glomerulonephritis, membranous nephropathy, minimal change nephrotic syndrome,
Goodpasture's syndrome, granulomatosis with polyangiitis, Graves' disease,
Guillain-Barre
syndrome, Hashimoto's thyroiditis, hemolytic anemia, immune hemolytic anemia,
Henoch-
Schonlein purpura (HSP), Herpes gestationis or pemphigoid gestationis (PG),
hypogammalglobulinemia, IgA Nephropathy, IgG4-related sclerosing disease,
Immune
thrombocytopenic purpura (ITP), Inclusion body myositis (IBM), Interstitial
cystitis (IC),
juvenile arthritis, juvenile diabetes (Type I diabetes), juvenile myositis
(JM), Kawasaki
disease, Lambert-Eaton syndrome, Leukocytoclastic vasculitis, Lichen planus,
Lichen
sclerosus, Ligneous conjunctivitis, Linear IgA disease (LAD), Lupus, Lyme
disease chronic,
Meniere's disease, Microscopic polyangiitis (MPA), Mixed connective tissue
disease
(MCTD), Mooren's ulcer, Mucha-Habermann disease, Multiple sclerosis,
Myasthenia gravis,
Myositis, Narcolepsy, Neuromyelitis optica, Neutropenia, Ocular cicatricial
pemphigoid,
Optic neuritis, Palindromic rheumatism (PR) PANDAS, Paraneoplastic cerebellar
degeneration (PCD), Paroxysmal nocturnal hemoglobinuria (PNH), Parry Romberg
CA 3050553 2019-07-25
syndrome, Pars planitis (peripheral uveitis), Parsonnage-Turner syndrome,
Pemphigus,
peripheral neuropathy, perivenous encephalomyelitis, Pernicious anemia (PA),
POEMS
syndrome, Polyarteritis nodosa, Polyglandular syndromes type I, II, III,
Polymyalgia
rheumatica, Polymyositis, Postmyocardial infarction syndrome,
Postpericardiotomy
syndrome, Primary biliary cirrhosis, Primary sclerosing cholangitis,
Progesterone dermatitis,
Psoriasis, Psoriatic arthritis, Pure red cell aplasia (PRCA), Pyoderma
gangrenosum,
Raynaud's phenomenon, Reactive arthritis, Reflex sympathetic dystrophy,
Relapsing
polychondritis, Restless legs syndrome (RLS), Retroperitoneal fibrosis,
Rheumatic fever,
Rheumatoid arthritis, Sarcoidosis, Schmidt syndrome, Scleritis, Scleroderma,
Sjogren's
syndrome, Sperm & testicular autoimmunity, Stiff person syndrome (SPS),
Subacute
bacterial endocarditis (SBE), Susac's syndrome, Sympathetic ophthalmia (SO),
Takayasu's
arteritis, Temporal arteritis/Giant cell arteritis, Thrombocytopenic purpura
(TTP), Tolosa-
Hunt syndrome (THS), Transverse myelitis, Type I diabetes, Ulcerative colitis
(UC),
Undifferentiated connective tissue disease (UCTD), Uveitis, Vasculitis,
Vitiligo, Wegener's
granulomatosis (or Granulomatosis with Polyangiitis (GPA)), idiopathic
thrombocytopenia
purpura, splenomegaly;
(xxiii) Systemic inflammatory response syndrome, cytokine release syndrome,
cytokine storm, immune reaction to a drug(s) or therapy(s), immune reaction to
an immune
activating drug(s) or agent(s) or treatment(s) or intervention(s), immune
reaction to
immunotherapy and/or immune-oncology and/or immunomodulatory drug(s) and/or
treatment(s), adverse reaction to adoptive T-cell therapy(s), adverse reaction
to a chimeric
antigen receptor T-cell therapy(s) (CAR-T cell therapy(s)), adverse reaction
to a immune
checkpoint inhibitor(s), adverse reaction to monoclonal antibody drug(s),
tumor lysis
syndrome;
(xxiv) cancer and Graft Versus Host Disease (GVHD) in transplantation therapy
in a
cancer patient;
(xxv) cardiovascular diseases and conditions associated with thrombosis and/or
the
formation of atherosclerotic plaques and/or ischemia and/or ischemic
conditions and/or
associated conditions including, without limitation, ischemia-reperfusion
injury, myocardial
ischemia, ischemic heart disease, chronic stable angina pectoris, first or
recurrent myocardial
infarction (MI), congestive heart failure, an acute coronary syndrome, muscle
cell damage,
necrosis, cardiac arrhythmia(s), non-Q wave MI, unstable angina, high blood
pressure,
coronary artery disease, coronary arterial thrombosis, ischemic hypoxia,
cyanosis, gangrene,
acute limb ischemia, stroke, ischemic stroke, brain ischemia, vascular
dementia, ischemic
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CA 3050553 2019-07-25
sudden death, transient ischemic attack (TIA), thrombophlebitis, ischemic
colitis, mesenteric
ischemia, angina pectoris, ischemic heart disease, ischemic neuropathy,
hypoxic-ischemic
encephalopathy, cerebral hypoxia, brain hypoxia, ischemia resulting from
vascular occlusion,
cerebral infarction, stroke and related cerebral vascular diseases (including
cerebrovascular
accident and transient ischemic attack), muscle cell damage, necrosis,
ventricular
hypertrophy, ventricular enlargement (including dilated cardiac myopathy and
heart failure),
Prinzmetal's angina, peripheral occlusive arterial disease (e.g., peripheral
arterial disease,
intermittent claudication, critical leg ischemia, prevention of amputation,
prevention of
cardiovascular morbidity such as MI, stroke or death), pericardial effusion,
constrictive
pericarditis, thrombosis, thrombotic or thromboembolic conditions, circulatory
disease
caused by blood clot (i.e. diseases involving fibrin formation, platelet
activation, and/or
platelet aggregation), thrombotic or thromboembolic symptoms of thromboembolic
stroke
(including that resulting from atrial fibrillation or ventricular mural
thrombus), arterial
cardiovascular thromboembolic disorders, venous cardiovascular thromboembolic
disorders,
thromboembolic disorders in the chambers of the heart, venous thrombosis
(including deep
vein thrombosis), arterial thrombosis, cerebral thrombosis, cerebral arterial
thrombosis,
pulmonary embolism, cerebral embolism, kidney embolism, arterial embolism,
thrombophilia, disseminated intravascular coagulation, restenosis, atrial
fibrillation,
atherosclerotic vascular disease, atherosclerotic plaque formation,
atherosclerosis,
atherosclerotic plaque rupture, peripheral arterial disease, coagulation
syndromes,
intermittent claudication, transplant atherosclerosis, vascular remodeling
atherosclerosis,
diabetic complications comprising retinopathy, nephropathy and neuropathy,
thromboembolic consequenses of surgery, interventional cardiology or
immobility,
thromboembolic consequenses of medication (such as oral contraceptives,
hormome
replacement and heparin), thrombotic consequenses of atherosclerotic vascular
disease and
atherosclerotic plaque rupture leading to tissue ischemia, prevention of
atherosclerotic plaque
formation, transplant atherosclerosis, thrombotic or thromboembolic
complications of surgery
including interventional cardiology, thromboembolic complications of pregancy
including
fetal loss, thromboembolic consequences of thrombophilia (e.g., Factor V
Leiden, and
homocystinenimia), prothrombotic consequences and/or complications of cancer,
prevention
of thrombosis on artificial surfaces (such as stents, blood oxygenators,
shunts, vascular access
ports, vascular grafts, artificial valves, etc.), coagulopathies (e.g.,
disseminated intravascular
coagulation), coagulation syndromes, vascular remodeling atherosclerosis,
restenosis and
systemic infection, Kasabach-Merritt syndrome, occlusion (e.g. after a bypass)
and
47
CA 3050553 2019-07-25
reocclusion (e.g., during or after percutaneous transluminal coronary
angioplasty),
thromboembolic disorders resulting from conditions including but not limited
to
atherosclerosis, surgery or surgical complications, prolonged immobilization,
arterial
fibrillation, congenital thrombophilia, cancer, diabetes, effects of
medications or hormones,
complications of pregnancy and thrombosis resulting from prosthetic valves or
other
implants, indwelling catheters, stents, cardiopulmonary bypass, hemodialysis,
or other
procedures in which blood is exposed to an artificial surface that promotes
thrombosis; or
(xxvi) diseases or disorders or conditions
treatable/ameliorated/prevented/combated
by conferring/maintaining blood vessel patency in a subject, which can be
useful during
interventional cardiology or vascular surgery including bypass grafting,
arterial
reconstruction, atherectomy, vascular graft and stent patency, organ, tissue
and cell
implantation and transplantation, preservation of host and/or graft tissue as
related to organ
transplantation,
wherein the method comprises administering to the subject an effective amount
of at
least one compound, or a composition containing at least one compound, which
reduces FiFo
ATP hydrolysis in a subject, or a pharmaceutically-acceptable salt, solvate,
hydrate or
prodrug thereof.
Another aspect is a method of treating, ameliorating, preventing or combating
any such
disease or disorder by administering to a subject in need thereof a
therapeutically effective
amount of a compound(s) and/or composition(s) as described herein.
Another aspect is the use of a compound(s) and/or composition(s) as described
herein for the
manufacture of a medicament for the treatment, amelioration, prevention or
combating of any
disease or disorder, optionally a disease or disorder mentioned or inferred
herein.
In some embodiments, the subject is further/also administered with one or more
compounds
or compositions approved for human use, optionally for anti-cancer use, by the
United States
Food and Drug Administration (FDA) and/or European Medicines Agency (EMA),
optionally in the same pharmaceutical composition.
In some embodiments, the mg/kg drug dose administered to the subject is
comparable with or
larger than the mg/kg dose which would be administered to a subject of smaller
bodily size
(optionally a subject of another, smaller, species), and optionally the mg/kg
dosage
48
CA 3050553 2019-07-25
administered to adult humans is comparable or greater than the No Observed
Adverse Effects
Level (NOAEL) mg/kg dosage in mice housed at 22 C. The latter would greatly
surprise
someone of the art because it is very distinct from most other drugs, wherein
bigger species
are administered much lower mg/kg doses, not comparable or larger.
BRIEF DESCRIPTION OF THE DRAWINGS
Figures 1, 2, 3, 4, 5, 6, 7, 8 show anti-cancer activity of carboplatin (10
M), BTB06584 (10
M), BTB06584 (100 M), BMS-199264 (10 M), BMS-199264 (100 M), compound 31
(10 M), almitrine dimesylate (at 10 and 100 M), compounds 6a and 6b (at 10
and 100
04), respectively in the NCI one-dose assay. Figure 9 consolidates data from
prior figures to
show that anti-cancer activity scales with inhibition of FIR:, ATP hydrolysis.
Figure 10 shows
anti-cancer activity of compounds 7a and 7b in the NCI one-dose (10 IA)
assay. Figure 11
shows anti-cancer activity of compounds 8a and 8b in the NCI one-dose (10 M)
assay.
Figure 12 consolidates data from prior figures to compare the anti-cancer
activities of
compounds 6a, 6b, 7a, 7b, 8a and 8b. Figure 13 shows anti-cancer activity of
BMS-199264,
Figure 14 shows anti-cancer activity of 31, Figure 15 shows anti-cancer
activity of 6a and 6b,
Figure 16 shows anti-cancer activity of 8a and 8b, in the NCI five-dose assay.
Figure 17
recasts data from Figures 8 & 15. Figure 18 interprets data from Figures 8,
15, and 17. Figure
19 interprets data from Figure 13. Figure 20 recasts data from Figure 16.
Figure 21 recasts
data from Figure 7. Figure 22 (A) shows that different cancer cell lines of
the NCI one dose
assay are tested with different numbers of cancer cells and (B) presents drug
aggregation data
for compound 7b. Figure 23 presents in vivo mouse data for compounds 6a and
6b. Figure 24
presents experimental data showing that IF1 is a determinant of lifespan.
Figure 25 shows
that, in vivo, inhibiting FIF0 ATP hydrolysis safely reduces the rate of
oxidative
phosphorylation and ROS generation, shown with forebrain neurons. Figure 26
shows that, in
vivo, inhibiting FiFo ATP hydrolysis safely reduces the rate of oxidative
phosphorylation,
shown with hepatocytes. Figure 27 shows that, in vivo, inhibiting FIE) ATP
hydrolysis safely
reduces the rate of oxidative phosphorylation, shown in intestine (colonic
cells). Figure 28 is
a diagram illustrating how decreasing [ROS] in a cell, for example by
inhibiting FiFo ATP
hydrolysis which reduces the oxidative phosphorylation/ROS generation rate,
can
prolong/increase the information fidelity of DNA, which slows/reverses aging.
Figure 29
presents some protein sequence embodiments of the invention. Figure 30
presents
experimental data on the racemization rate of 6b. Figure 31 discloses that
invention
compounds can reduce FIFO ATP synthesis by uncoupling the proton motive force,
with
49
CA 3050553 2019-07-25
supporting experimental data. Figure 32 discloses spectral data for compounds
6a and 6b.
Figure 33 discloses spectral data for compounds 7a and 7b. Figure 34 compares
spectral data
from Figures 32 and 33. Figure 35 discloses spectral data for compounds 8a and
8b. Figure
36 discloses spectral data for compound 31. Figure 37 discloses spectral data
for the
compound synthesized by Scheme IHa of this disclosure.
DETAILED DESCRIPTION OF THE INVENTION
Some definitions
As used herein with reference to the utilities described, the terms "treating"
or "treatment"
encompass both responsive and prophylaxis/preventative measures designed to
inhibit or
delay the onset of the disease or disorder, or to cure/eradicate, alleviate,
palliate, reverse,
prevent, ameliorate, lessen, reduce, modulate, stabalize, delay, suppress,
manage, reduce
predisposition to, reduce risk of, prevent, reduce reoccurrence of, lengthen
time to remission
of, or slow progression/spread of the disease or disorder and/or one or more
of its symptoms
and/or increase quality/length of life and/or improve subject
outcome/wellness. The terms
"subject" and "patient" refer to organisms to be treated by the
compounds/methods of the
present invention and can refer to a human or animal. As used herein, the term
"therapeutically effective amount" or "effective amount" refers to the amount
of a compound
(e.g. a compound of the present disclosure) sufficient to effect a
therapeutically/cosmetically/aesthetically beneficial/desired result
including, for example,
mitigating (reducing frequency/duration/severity) or eliminating one or more
symptoms of
the disease/disorder/condition/sub-optimum, or treating at least one
physiological defect or
pathology or etiology that causes or contributes to the
disease/disorder/condition/sub-
optimum being treated. In the case of cancer, a therapeutically effective
amount can be, for
example that which slows/halts/stabalises/regresses cancer
proliferation/spread/invasion/malignancy/danger in the subject and/or which
slows/halts/stabalises/regresses cancer associated cachexia. A therapeutically
effective
amount accounts for treatment variables including, for example, dose,
duration, timing and
route of administration. An invention embodiment is to administer a
compound(s) of this
invention to a subject diagnosed with cancer, suspected of having cancer,
exhibiting
symptoms of a cancer, at risk of cancer (e.g., a human who is genetically or
otherwise
predisposed to developing a cancer), susceptible to cancer,
recovering/recovered from cancer
or free of cancer. Palliative use of a compound(s) of this invention,
optionally in a subject
with cancer, is contemplated by, and componentry to, this invention.
CA 3050553 2019-07-25
Cancer is herein used to mean any member of a class of diseases/disorders
characterized by
uncontrolled/undesirable/abnormal/dysregulated/unregulated, including
harmful/dangerous
(to health and/or lifespan), division of cells, including that independent of
normal regulatory
.. mechanisms (e.g. loss of contact inhibition). A "tumor" comprises one or
more cancerous
cells. Cancer cells, in some cases, gain the ability to invade other tissues,
either by direct
growth into adjacent tissue through invasion or by implantation into distant
sites by
metastasis. Metastasis is defined as the stage in which cancer cells are
transported through the
bloodstream or lymphatic system. The cancer may be, for illustrating example,
a solid tumor,
metastatic cancer, non-metastatic cancer, malignant cancer, benign cancer or
pre-cancer. In
some embodiments, the cancer may be a chemo-resistant or multidrug resistant
cancer, i.e. a
refractive form of cancer. It should be appreciated that a
composition/compound of the
invention may be used alone or in combination with one or more additional anti-
cancer
agents or treatments (e.g. chemotherapeutic agents, targeted therapeutic
agents, pseudo-
.. targeted therapeutic agents, hormones, radiation, surgery, etc., or any
combination of two or
more thereof), optionally a further composition(s)/compound(s) of the
invention. In some
embodiments, a composition(s)/compound(s) of the invention may be administered
to a
subject who has undergone a treatment involving one or more of surgery,
radiation,
chemotherapy. In certain embodiments, a composition or compound of the
invention may be
.. administered chronically to prevent, or reduce the risk of, a cancer
recurrence. According to
one embodiment, the subject to be treated is characterized by the presence of
a precancerous
condition, and the administering of the compound is effective to prevent
development of the
precancerous condition into the cancerous condition. This can occur by
destroying the
precancerous cell prior to or concurrent with its further development into a
cancerous state.
For the purpose of this invention, beneficial or desired results in the
subject include, but are
not limited to, alleviation of symptoms, diminishment of extent of disease,
stabilized (i.e., not
worsening) state of disease, delay or slowing of disease progression,
amelioration or
palliation of the disease state, remission (partial or total), disease
prevention, or reducing
predisposition to the disease, prolonging survival as compared to expected
survival if not
receiving treatment. In some embodiments, a compound(s) of the present
invention is used to
prevent the growth of a tumor or cancer, and/or prevent the metastasis of a
tumor or cancer,
and/or to shrink or destroy a cancer and/or treat complications of cancer. A
treatment using
one or more of the disclosed therapeutic compounds and compositions disclosed
herein may
decrease the growth rate of tumor cells, decrease the cell division rate of
tumor cells,
51
CA 3050553 2019-07-25
decrease the extent of invasion of tumor cells into adjacent tissue or organs,
decrease the
extent of metastasis, decrease angiogenesis, increase apoptosis, increase
tumor cell death,
increase tumor cell necrosis, or all or any combination thereof. A treatment
using one or more
of the disclosed therapeutic compounds and compositions disclosed herein may
decrease
hyperplasia, decrease the growth rate of hyperproliferating cells, decrease
the cell division
rate of hyperproliferating cells, decrease the extent to which
hyperproliferating cells becomes
cancerous, decrease angiogenesis, decrease nodule formation, decrease cyst
formation,
increase apoptosis, increase tumor cell death and/or increase tumor cell
necrosis, or all or any
combination thereof.
Jurisdictions
To simultaneously capture the claim format of both Europe ("use") and USA
("method") and
other legal jurisdictions in a single disclosure, any place that one of the
following terms
(herein specified inside a ") is used, then also componentry to this
disclosure is any of the
other terms in this sentence, inside ", substituted in its place (including
iterated
substitutions): "the use of', "the method of using", "the method of using
(administering an
effective amount to a subject)", the method of using (administering an
effective amount
to a subject in need thereof)", "use of', "method of use of', "use", "method
of use", "method"
"for use in", "for use in the", "for use according to", "for use in a method
of', for use in a
method (administering an effective amount to a subject) of', "for use in a
method
(administering an effective amount to a subject in need thereof) of', "for use
for", "for use in
a method for", "for use in a method of', "for use in", "to use", "a method of
using", "a method
(administering an effective amount to a subject in need thereof) of using",
"used to", "used in
a method of', "used in a method (administering an effective amount to a
subject in need
thereof) of', "in use as", "in a method of use as", "in a method
(administering an effective
amount to a subject) of use as", "in a method (administering an effective
amount to a subject
in need thereof) of use as", "for a method of use for", "for a method of use
of', "for a method
of', "for a method (wherein an effective amount of the compound or compounds
is
administered to the subject) of", "used", "used in a method", "use", "use a
method of',
"method of use", "a method of use", "method", "a method", "use in a method",
"use in a
method for", "use in a method of', "use in a method of treatment", "use in a
method of
treating", "to use", "a method of using", "for use in", "for a method of use
in", "use for the
manufacture of a medicament", "use for the manufacture of a medicament for",
"use for the
manufacture of a medicament to", "for use in the manufacture of a medicament",
"for use in
52
CA 3050553 2019-07-25
the manufacture of a medicament to", "for use in the manufacture of a
medicament for", "for
use in a method of manufacture of a medicament", for use in a method of
manufacture of a
medicament for", "for a method of manufacture of a medicament", "for
manufacture of a
medicament". If this simultaneous capture technically/gramatically fails in a
place, places or
entirity, then someone of the art will recognize my intent by this paragraph
and factor in the
necessary correction(s) to arrive at a technically/gramatically correct
specification/interpretation for that jurisdiction. In some embodiments, where
the word
"method" is used in a claim or statement in this disclosure, it is substituted
with "process" or
"method (administering an effective amount to a subject)" or "method
(administering an
effective amount to a subject in need/want thereof)" or "method (administering
an effective
amount to a subject in need of treatment)" or "method of treatment". In some
embodiments,
where the word "subject" is used in a claim or statement in this disclosure,
it is substituted
with "subject in need of treatment" or "subject in need thereof' or "subject
in need/want
thereof'. In some embodiments, where the word "effective" is used in a claim
or statement in
this disclosure, it is substituted with "therapeutically effective". In some
embodiments, where
the word "manufacture" is used in a claim or statement in this disclosure, it
is substituted with
"preparation" or "production" or "import" or "transport" or "import across a
national
boundry" or "import into the country" or "sale" or "distribution" or "supply"
or "offer to
supply" or "advertisment" or "handling" or "prescription" or "testing" or
"trialling" or
"administration" or "administer". As known by those of the art, there are 3
different claim
types: method of medical treatment, Swiss-type and Product by process (purpose-
limited-
product format, EPC 2000) claims: in this disclosure, when a claim or
statement is given in
one of these forms it also incorporates by reference the same subject matter
in both the other
claim forms, wherein it is straightforward for one of the art to convert
subject matter in one of
these claim forms into both the other claim forms. Also, incorporated by
reference is subject
matter of this disclosure written in other claim forms that have the same
intent as claim forms
herein.
A pharmaceutical composition comprising at least one (optionally more than
one) compound,
as described herein, and a pharmaceutically-acceptable carrier or excipient or
diluent. A
pharmaceutical composition comprising at least one (optionally more than one)
compound, as
described herein, or a pharmaceutically-acceptable salt, solvate, hydrate or
prodrug thereof,
optionally a compound(s) of one or more of Formula (I), (II), (III), (IV),
(V), (VI), (VII), [X],
optionally a compound(s) that reduces FIFO ATP hydrolysis in a subject, and/or
a
53
CA 3050553 2019-07-25
composition containing at least one compound defined herein, and a
pharmaceutically-
acceptable carrier or excipient or diluent.
Use of a compound, specified herein, for treatment of a disease, specified
herein. Use of a
compound(s) defined herein, and/or use of a composition containing at least
one compound
defined herein, and/or use of a pharmaceutical composition defined herein, for
treatment of
one or more diseases or disorders specified herein. Use of a compound(s)
defined herein, or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, and/or
use of a
composition containing at least one compound defined herein, and/or use of a
pharmaceutical
composition defined herein, optionally a compound(s) and/or composition(s)
that reduces
FiFo ATP hydrolysis in a subject, optionally a compound(s) of one or more of
Formula (I),
(II), (III), (IV), (V), (VI), (VII), [X], for
treatment/amelioration/prevention/reversal/combat of
one or more diseases or disorders or unwanted/undesirable physiological
processes (or
consequences) or an unwanted/undesirable aesthetic specified herein.
Use of a compound(s) specified herein for the manufacture of a medicament. Use
of a
compound, specified herein, for manufacture of a medicament for treatment of a
disease,
specified herein. Use of a compound(s) defined herein, and/or use of a
composition
containing at least one compound defined herein, and/or use of a
pharmaceutical composition
defined herein, for the manufacture of a medicament for treatment of one or
more diseases or
disorders specified herein. Use of a compound(s) defined herein, or a
pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof, and/or use of a
composition containing at
least one compound defined herein, and/or use of a pharmaceutical composition
defined
herein, optionally a compound(s) and/or composition(s) that reduces F 1 Fo ATP
hydrolysis in
.. a subject, optionally a compound(s) of one or more of Formula (I), (II),
(III), (IV), (V), (VI),
(VII), [X], for the manufacture of a medicament for
treatment/amelioration/prevention/reversal/combat of one or more diseases or
disorders or
unwanted/undesirable physiological processes (or consequences) or an
unwanted/undesirable
aesthetic specified herein.
A compound(s) and/or composition(s) described herein for use in a method of
treatment of
the human or animal body by therapy.
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CA 3050553 2019-07-25
A compound(s) and/or composition(s) described herein for use in a method of
treatment/improvement/enhancement of the human or animal body by therapy.
A method of treating, ameliorating, preventing or combating a disease or
disorder by
administering a therapeutically effective amount to the subject of at least
one compound as
defined herein. A method of treating, ameliorating, preventing or combating a
disease or
disorder by administering a therapeutically effective amount to the subject in
need thereof of
at least one compound defined as herein. A method of treating, ameliorating,
preventing or
combating a disease or disorder by administering to a subject in need of
treatment an
effective amount of at least one compound defined herein. A method of
treating/ameliorating/preventing/reversing/combating one or more of a
disease/disorder or
unwanted/undesirable physiological process (or its consequences) or an
unwanted/undesirable aesthetic, specified herein, in a subject wherein the
method comprises
administering an effective amount to the subject of at least one compound
defined herein, or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
optionally a
compound(s) of one or more of Formula (I), (II), (III), (IV), (V), (VI),
(VII), [X], optionally a
compound(s) that reduces FiFo ATP hydrolysis in a subject, and/or a
composition containing
at least one compound defined herein, and/or a pharmaceutical composition
defined herein. A
method of treating/ameliorating/preventing/reversing/combating one or more of
a
disease/disorder or unwanted/undesirable physiological process (or its
consequences) or an
unwanted/undesirable aesthetic, specified herein, in a subject wherein the
method comprises
administering an effective amount to the subject in need/want thereof of at
least one
compound defined herein, or a pharmaceutically-acceptable salt, solvate,
hydrate or prodrug
thereof, optionally a compound(s) of one or more of Formula (I), (II), (III),
(IV), (V), (VI),
(VII), [X], optionally a compound(s) that reduces FiFo ATP hydrolysis in a
subject, and/or a
composition containing at least one compound defined herein, and/or a
pharmaceutical
composition defined herein.
Use of a compound(s) defined herein, or a pharmaceutically-acceptable salt,
solvate, hydrate
or prodrug thereof, and/or use of a composition containing at least one
compound defined
herein, and/or use of a pharmaceutical composition defined herein, optionally
a compound(s)
and/or composition(s) that reduces FIR) ATP hydrolysis in a subject,
optionally a
compound(s) of one or more of Formula (I), (II), (III), (IV), (V), (VI),
(VII), [X], for
treatment/amelioration/prevention/reversal/combat of one or more diseases or
disorders or
CA 3050553 2019-07-25
unwanted/undesirable physiological processes (or consequences) or an
unwanted/undesirable
aesthetic specified herein, in subject, wherein the administration to the
subject is topical/local
(not systemic). A method of
treating/ameliorating/preventing/reversing/combating one or
more of a disease/disorder or unwanted/undesirable physiological process (or
its
consequences) or an unwanted/undesirable aesthetic, specified herein, in a
subject, wherein
the method comprises topically/locally (not systemically) administering an
effective amount
to the subject in need/want thereof of at least one compound defined herein,
or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
optionally a
compound(s) of one or more of Formula (I), (II), (III), (IV), (V), (VI),
(VII), [X], optionally a
compound(s) that reduces FiFo ATP hydrolysis in a subject, and/or a
composition containing
at least one compound defined herein, and/or a pharmaceutical composition
defined herein,.
Use of a compound(s) defined herein, or a pharmaceutically-acceptable salt,
solvate, hydrate
or prodrug thereof, and/or use of a composition containing at least one
compound defined
herein, and/or use of a pharmaceutical composition defined herein, optionally
a compound(s)
and/or composition(s) that reduces FIFO ATP hydrolysis in a subject,
optionally a
. compound(s) of one or more of Formula (I), (II), (III), (IV), (V), (VI),
(VII), [X], for
treatment/amelioration/prevention/reversal/combat of one or more diseases or
disorders or
unwanted/undesirable physiological processes (or consequences) or an
unwanted/undesirable
aesthetic specified herein, in a subject, wherein the subject is also
administered with one or
more compounds or compositions approved for human use, optionally for anti-
cancer use, by
the United States Food and Drug Administration (FDA) and/or European Medicines
Agency
(EMA), optionally in the same pharmaceutical composition. A method of
treating/ameliorating/preventing/reversing/combating one or more of a
disease/disorder or
unwanted/undesirable physiological process (or its consequences) or an
unwanted/undesirable aesthetic, specified herein, in a subject, wherein the
method comprises
administering an effective amount to the subject in need/want thereof of at
least one
compound defined herein, or a pharmaceutically-acceptable salt, solvate,
hydrate or prodrug
thereof, optionally a compound(s) of one or more of Formula (I), (II), (III),
(IV), (V), (VI),
(VII), [X], optionally a compound(s) that reduces FiFo ATP hydrolysis in a
subject, and/or a
composition containing at least one compound defined herein, and/or a
pharmaceutical
composition defined herein, wherein the subject is also administered with an
effective
amount (which can be less than when administered alone) of one or more
compounds or
compositions approved for human use, optionally for anti-cancer use, by the
United States
56
CA 3050553 2019-07-25
Food and Drug Administration (FDA) and/or European Medicines Agency (EMA),
optionally in the same pharmaceutical composition.
Use of a compound(s) defined herein, or a pharmaceutically-acceptable salt,
solvate, hydrate
or prodrug thereof, and/or use of a composition containing at least one
compound defined
herein, and/or use of a pharmaceutical composition defined herein, optionally
a compound(s)
and/or composition(s) that reduces FIFO ATP hydrolysis in a subject,
optionally a
compound(s) of one or more of Formula (I), (II), (III), (IV), (V), (VI),
(VII), [X], for
treatment/amelioration/prevention/reversal/combat of one or more diseases or
disorders or
unwanted/undesirable physiological processes (or consequences) or an
unwanted/undesirable
aesthetic specified herein, in a subject, wherein comparable or larger mg/kg
dose is used in
larger subjects/animals/species, which is very distinct from most drugs, and
optionally the
mg/kg dosage administered to adult humans is comparable or greater than the No
Observed
Adverse Effects Level (NOAEL) mg/kg dosage in mice housed at 22 C. A method of
treating/ameliorating/preventing/reversing/combating one or more of a
disease/disorder or
unwanted/undesirable physiological process (or its consequences) or an
unwanted/undesirable aesthetic, specified herein, in a subject, wherein the
method comprises
administering an effective amount to the subject in need/want thereof of at
least one
compound defined herein, or a pharmaceutically-acceptable salt, solvate,
hydrate or prodrug
thereof, optionally a compound(s) of one or more of Formula (I), (II), (III),
(IV), (V), (VI),
(VII), [X], optionally a compound(s) that reduces FIF0 ATP hydrolysis in a
subject, and/or a
composition containing at least one compound defined herein, and/or a
pharmaceutical
composition defined herein, wherein a comparable or larger mg/kg compound(s)
dose is used
in, or administered to, a larger subject(s)/animal(s)/species, which is very
distinct from most
drugs, and optionally the mg/kg dosage administered to one or more adult
humans is
comparable or greater than the No Observed Adverse Effects Level (NOAEL) mg/kg
dosage
in mice housed at 22 C.
Invention
The invention of this disclosure hinges on the discovery, disclosed herein,
that some cancers
rely on FIFO ATP hydrolysis, even under normoxia (indeed under hyperoxia: ¨21%
02),
during some or all of their cell cycle. Evidence herein: compounds of this
disclosure, which
specifically inhibit FIFO ATP hydrolysis, slow cancer proliferation at
concentrations that they
do not harm normal cells.
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CA 3050553 2019-07-25
In some of the most dangerous cancers, refractory to present [chemo/radio]
therapies, during
some or all of their cell cycle, reactive oxygen species (ROS) decrease
[NADPH], because
NADPH is consumed in ROS mitigation processes, and this then pulls through
increased
pentose phosphate pathway (PPP) and glycolytic flux. But such a pivotal
increase in
glycolytic/PPP flux can only occur because of FiFo ATP hydrolysis, a
distinctive feature to
these cancers, which stops ATP produced by glycolysis from accumulating and
slowing
glycolysis by negative feedback inhibition of key glycolytic enzymes. This
increased PPP
flux maintains [NADPH] and ROS mitigation. In this way, these cancers can
maintain a very
high ROS mitigation capability, maintain very low intracellular EROS], and
tend to be the
most resistant to conventional [chemo/radio] therapies, which work, or often
don't work (!),
by increasing [ROS]. Compounds of this disclosure undermine this
process/resistance. By
inhibiting FIFO ATP hydrolysis, they increase the anti-cancer efficacy of any
chemical or
treatment that increases reactive oxygen species (ROS) in cancer cells. An
embodiment of
this disclosure is any such co-treatment(s). Indeed, a compound(s) of this
disclosure increases
the success rate of standard of care [chemo/radio] therapies and optionally
permits their use at
lower dosing, which reduces their horrendous side-effects. This disclosure
encompasses a
compound(s) of this invention, for example a compound(s) of Formula (I), (II),
(III), (IV),
(V), (VI), (VII) in co-therapy with one or more of surgery, chemotherapy,
immunotherapy,
immuno-oncology, radioimmunotherapy, biological therapy, hormone therapy,
radiotherapy
or any US Food and Drug Administration (FDA) and/or European Medicines Agency
(EMA)
approved drug(s) or treatment, for example, a drug/treatment approved for
cancer therapy. In
some embodiments the anti-cancer activity of a compound(s) of this invention,
for example a
compound(s) of Formula (I), (II), (III), (IV), (V), (VI), (VII), synergises
with (potentiates) the
anti-cancer activity of an FDA and/or EMA approved anti-cancer treatment(s)
e.g. one or
more of chemotherapy, radiotherapy, immunotherapy, surgery etc. In other
words, their
combined anti-cancer effect is greater than simply being the sum of each
alone. In some
embodiments, a compound(s) of this invention is used as an adjuvant or
neoadjuvant to
another cancer treatment(s) e.g. used as an adjuvant or neoadjuvant to chemo
and/or
radiotherapy and/or surgery. In some embodiments a compound(s) of this
invention, for
example a compound(s) of Formula (I), (II), (III), (IV), (V), (VI), (VII),
makes a cancer(s)
more radiosensitive/less radioresistant and/or more chemosensitive/less
chemoresistant i.e.
more amenable to treatment by radio- and/or chemo- therapy, acts as a
radiosensitizer and/or
chemosensitizer. This is very valuable for treating radio- and/or chemo-
resistant cancers.
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CA 3050553 2019-07-25
Chemotherapies are well known to those of the art, including, but not limited
to, cisplatin,
carboplatin, taxol, oxaliplatin etc, and tend to be (very) toxic. Encompassed
herein is a
method of reducing, treating and/or preventing adverse or undesired effects
associated with
conventional therapy including, but not limited to, chemotherapy,
radiotherapy,
immunotherapy, wherein a compound(s) provided herein, e.g., a compound(s) of
Formula (I),
(II), (III), (IV), (V), (VI), (VII) or a pharmaceutically acceptable salt,
solvate, hydrate,
prodrug thereof, is administered to a subject prior to, during, or after the
occurrence of the
adverse effect associated with conventional therapy, optionally wherein the
dosage/frequency/use of the conventional therapy is decreased.
In other embodiments, a compound(s) of this disclosure is used as cancer
therapy alone.
Indeed, this is a much more cancer-targeted therapeutic approach. The most
dangerous
cancers use this distinctive metabolism, with ATP synthase distinctively in
reverse,
consuming glycolytic ATP, to yield high glycolytic rate, thence abundant
glycolytic
intermediates for biosynthesis and, crucially, to keep [ROS] low (as prior
disclosed), which is
necessary to cancer immortality ("limitless replicative potential", a Hallmark
of cancer [26])
and thence danger. This distinction is targeted, by compound(s) of this
disclosure, without
significant damage to normal cells. Normal adult cells normally use a
different metabolism,
with ATP synthase more in forward mode, and a higher ATP yield from glucose,
but at the
cost of higher [ROS] and mortality.
This reliance of normal cells upon the forward mode of ATP synthase makes them
exquisitely susceptible to oligomycin. The compounds of this disclosure are
useful for anti-
cancer treatment, unlike oligomycin, because of their distinction from
oligomycin, which
couldn't have been foreseen without the inventive steps of this disclosure. In
normal cells that
are actively respiring (known as state 3 respiration [3]), inhibitors of the
forward mode of
ATP synthase (e.g. oligomycin) cause a state 3 to state 4 transition,
hyperpolarize
decrease 02 consumption and reduce [ATP] (so called "modulators" of the
forward mode of
ATP synthase, e.g. Bz-423, can also cause one or more of these effects) whilst
a specific
inhibitor of the reverse mode of ATP synthase does not exert these effects at
a working
concentration ([12-13], herein incorporated in their entirety). However, at
this working
concentration, after inhibition of the respiratory chain (e.g. blocked by
rotenone, or some
other respiratory chain inhibitor, or by a reduced 02 concentration), a
specific inhibitor of the
reverse mode of ATP synthase will depolarise Tim. This feature distinguishes a
molecule that
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CA 3050553 2019-07-25
inhibits the reverse mode of ATP synthase significantly more than it
inhibits/modifies the
forward mode of ATP synthase, and/or inhibits/modifies ATP synthesis. Such a
molecule, put
into use as an anti-cancer therapeutic, is an embodiment of this invention. A
further
embodiment is the process/method of seeking new anti-cancer molecules by
assaying
whether a candidate molecule can depolarise Tim, when Tim is maintained by
FiFo ATP
hydrolysis (e.g. when OXPHOS is blocked by a respiratory chain inhibitor or
insufficient 02),
but that can't hyperpolarize Tim and/or decrease 02 consumption, when Tim is
maintained by
proton pumping by complexes of the respiratory chain. If a candidate molecule
meets these
requirements, it is an anti-cancer therapeutic, as determined by the invention
of this
disclosure.
Some cancers intrinsically rely upon ATP synthase in reverse, as revealed by
experimental
data of this disclosure, and further cancers can have this reliance imposed
upon them, to
maintain Tim in the hypoxia of a solid tumour, which also makes them
susceptible to drugs of
this disclosure. Significant lactate release is correlated with the most
dangerous cancers and
poor patient outcomes (numerous studies find this: example: [27]). High
lactate release
indicates high glycolytic rate, which FIFO ATP hydrolysis enables, and which
drugs of this
disclosure attack. This invention confronts the most deadly cancers by
discovering/disclosing
a cancer-specific weakness, and the means to selectively attack it.
All the following molecules are ¨ in use as anti-cancer therapeutics -
embodiments of this
invention: (1) Molecules that inhibit the reverse, and not the forward, mode
of ATP synthase,
(2) Molecules that inhibit the reverse more than than forward mode of ATP
synthase, (3)
Molecules that inhibit the reverse mode of ATP synthase, and not its forward
mode, but that
shuttle protons across the mitochondrial inner membrane, dissipating the pmf
as heat
(uncoupling [3]), which reduces FiFo ATP synthesis, and in a further
embodiment:
uncoupling molecules that reduce FIR' ATP hydrolysis more than FIF0 ATP
synthesis, (4)
Molecules that inhibit ATP hydrolysis more than ATP synthesis at the
mitochondrial inner
membrane, (5) Molecules that have a lower IC50 or ECso for FiFo ATP hydrolysis
than Fi Fo
ATP synthesis. This invention discloses the process/method of using one or
more molecular
species, each with one or more of the characteristics in the aforementioned
numbered points,
as an anti-cancer medicine or treatment. Some examples are presented in this
disclosure. Any
cancer therapy or treatment or drug that leverages, relies upon, utilises or
targets that cancers
employ ATP synthase in its reverse mode is an embodiment of this disclosure.
CA 3050553 2019-07-25
MECHANISTIC DISTINCTION FROM POLYICETIDE FiFo ATP SYNTHASE
INHIBITORS
The compounds of this invention act by a distinctly different mechanism, upon
cancer cells,
than oligomycin. Drugs that act against the same molecular target have a
similar pattern of
activity against the different cancer cell lines of the NCI-60 assay i.e. the
smaller, and the
larger, of their GI50 values are against the same cell lines (GI50 is compound
concentration
that causes 50% growth inhibition of a cell line relative to no-drug control).
The degree of
(dis)similarity can be measured using the COMPARE algorithm [28-29], which
employs a
Pearson correlation coefficient. For example, [30] found that the COMPARE
algorithm can
successfully group different FDA-approved anti-cancer drugs by their method of
action using
their NCI-60 GI50 data. Oligomycin A (NSC: 717694 [16]) inhibits FIFO ATP
synthase [4,
14] and so do other polyketides: cytovaricin (NSC: 349622 [16]), ossamycin
(NSC: 76627
[16]) and peliomycin (NSC: 76455 [161); indeed, their NCI-60 pattern responses
(GI50
values) correlate with that of oligomycin A: 0.896, 0.809 and 0.865
respectively (COMPARE
algorithm output, all significant at p <0.05). However, the NCI-60 pattern
response (GI50
values) of BMS-199264 is uncorrelated to that of oligomycin A (0.009). As is
that of
compound 31 (-0.009). As is that of compounds 8b (0.144, not significant at p
< 0.05) and 8a
(0.282, p= 0.03, so actually significant at p <0.05), which have very
correlated anti-cancer
activity with each other (0.752, p <0.00001). As is that of compounds 6b
(0.198, not
significant at p < 0.05) and 6a (0.207, not significant at p <0.05), which are
separated
stereoisomers of racemate 19a that both epimerize towards being the 19a
racemate during
NCI testing, and that have very correlated anti-cancer-activity (0.754, p
<0.00001). This
mechanistic distinction from oligomycin is vital because polyketide FIR ATP
synthase
inhibitors are poisonous to normal cells [15], which means they fail in cancer
xenograft
mouse experiments [16] and are without clinical utility.
Higher HIF-la (and lower pyruvate kinase (liver isoenzyme}, lower aspartate
aminotransferase 2 {mitochondrial} and lower ATP synthase) gene expressions
are reported
to be a marker of the Warburg effect [14] and correlate (at p < 0.05) with
insensitivity to the
polyketide FIN ATP synthase inhibitor, cytovaricin (Table 1 of [14]). By
contrast, using the
same cell lines and gene expression data set used to make Table 1 of [14], BMS-
199264
sensitivity (GI50) does not correlate (at p < 0.05) with any of these gene
expressions. Nor
does compound 31 or 6b or 8b sensitivity (G150, at p < 0.05). Figure 5 of [14]
presents
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CA 3050553 2019-07-25
apoptolidin resistant NCI-60 cell lines, resistant because they utilise the
Warburg effect [14],
but the majority of these cell lines are more sensitive to BMS-19264 than the
average, with a
lower GI50 value than the average GI50 value (3.9 M) for BMS-199264, half of
these cell
lines are more sensitive to compound 31 than the average, with a lower GI50
value than the
average GI50 value (13.2 j.iM) for 31, 37.5% of these cell lines are more
sensitive to
compound 6b than the average, with a lower G150 value than the average GI50
value (0.446
11M) for 6b.
The lower the bioenergetic cellular index (BEC) of a cancer cell [18], the
more it
demonstrates the Warburg effect and the more it relies on glycolytic rather
than oxidative
metabolism. BEC is, by one measure [19], the ratio amount of the 13 subunit of
Fl ATPase (p
¨Fl-ATPase; gene: ATP5B) to that of Glyceraldehyde 3-phosphate dehydrogenase
(GAPDH). I calculated BEC for the same cell lines analysed for Table 1 of
[14], using the
mRNA transcript amounts of ATP5B and GAPDH in each cell line, data sourced
from [31-
32], and then calculating the GATP5BMGAPDH] transcript ratio) for each of
these cancer
cell lines. Using transcript data rather than protein data is a limitation,
but [33] report that a
protein's cellular amount is generally well correlated (0.76) to its mRNA
transcript amount,
at least for cells in the NCI-60 assay, for the protein subset they studied.
And furthermore,
[14] relied on transcript data, so best comparison with [14] is made using
such data.
Polyketide FiFo ATP synthase inhibitors don't work well against cancer cells
exhibiting the
Warburg effect [14] and, indeed, for the cell lines analysed (same ones used
as for Table 1 in
[14]) there is a significant (at p <0.05) negative Pearson correlation between
logio(GI50) and
BEC for oligomycin A (-0.9411; oligomycin A logio(GI50) values sourced from
its entry
[NSC: 717694] in the "Query drug data" component of the CellMiner database
[32]). So, this
correlation shows that the more a cancer uses Warburg metabolism, the less its
danger is
mitigated by oligomycin A. This significantly reduces the utility of
oligomycin A as a cancer
medicine because a low BEC score (indicating Warburg metabolism) is
characteristic to some
of the most dangerous cancers, with the worst patient outcomes [18-20]. By
contrast, there is
no significant (at p <0.05) Pearson correlation between cancer cell line BEC
and GI50 values
for BMS-199264 (0.3639), 31 (0.4247) or 6b (0.0298) or 8b (-0.1388). This
means that,
distinctly from the polyketide FIFO ATP synthase inhibitors, their anti-cancer
action is not
restricted to those, often less dangerous, cancers that don't utilise Warburg
metabolism.
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CA 3050553 2019-07-25
Molecules of this disclosure undermine cancer by inhibiting the reverse mode
of ATP
synthase. It is true that polyketide FiFo ATP synthase inhibitors also inhibit
this mode, but
distinctly, in addition, they also inhibit the forward mode of ATP synthase,
indeed more
potently [II], and whilst they can exert anti-cancer activity, because this
forward mode is
vital to many cancers, it is also vital to many normal cells. This makes
polyketide FiFo ATP
synthase inhibitors unsuitable as clinical molecules. Molecules of this
disclosure are
therapeutic because of their distinction from, not their similarity to,
polyketide FIN ATP
synthase inhibitors.
[14] sum up with "Many cancer cells maintain a high level of anaerobic carbon
metabolism
even in the presence of oxygen, a phenomenon that is historically known as the
Warburg
effect. From our results, we conclude that macrolide inhibitors of the
mitochondrial FoFi-
ATP synthase selectively kill metabolically active tumor cells that do not
exhibit the Warburg
effect". So, [14] find that these macrolides only kill cancers reliant upon
OXPHOS, so using
FIFO-ATP synthase in its forward mode to generate ATP (which unfortunately is
also the
metabolic profile of many key types of normal cell) and thus macrolide
inhibition of the
forward mode of FiFo-ATP synthase is key to this (unspecific) anti-cancer
activity. By
contrast, the molecules of this disclosure exert anti-cancer activity by
inhibition of the reverse
mode of ATP synthase. BMS-199264 [4, 7, 9, 10, 11], BTB06584 [13], 31 [8] and
the
stereoisomer 6b (and its racemate, 19a) [5, 6] have been described previously,
as molecules
that can inhibit this mode, and this invention discloses their utility as anti-
cancer therapeutics,
with supporting experimental data, thence identifying new cancer drugs and,
more
fundamentally/importantly, a new cancer specific drug target: FIFO-ATP
hydrolysis (Figure
9). The opposite stereoisomer to 6b, 6a, also exerts anti-cancer activity
(Figure 8) because
both 6a and 6b racemize towards being the racemate, 19a, during NCI testing.
BTB06584
(100 04) exerts anti-cancer activity (Figure 3), despite not inhibiting FiFo-
ATP synthesis, as
a function of inhibiting FiFo-ATP hydrolysis (at >100 M), and critically it
isn't harmful to
normal cells (mouse cortical neurons) at this concentration [13]. Its anti-
cancer potency (none
at 10 M, observed at 100 M; Figures 2 and 3 respectively) matches its
inhibitory potency
for FiFo-ATP hydrolysis (none at 10 M, requires ?100 M [13]). BMS-199264 (10
M)
exerts anti-cancer activity (Figure 4). It doesn't harm normal cells (ex vivo
rat heart) at this
concentration [11]. In NCI five-dose testing [34-35], the mean GI50 for BMS-
199264 is 3.9
M (Figure 13), which is lower/better than 62% of the 102 FDA approved cancer
drugs in
[30], their mean GI50 values sourced from Table 1 of [30]: all are directly
comparable
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CA 3050553 2019-07-25
because they too are sourced from the NCI-60 five-dose assay. Furthermore, in
NCI five-dose
testing, the mean G150 for 6a and 6b is 0.666 tiM and 0.446 AM respectively
(Figure 15),
which, in the 6b case, is lower/better than 77% of 102 FDA approved cancer
drugs in [30],
including cisplatin (mean GIs() = 1.4 M), which is one of the most used, if
not the most used,
.. chemotherapies of today, but typically with terrible side-effects. A salt
of 6b, e.g. 6b HC1, is
likely to have an even lower GIso in NCI five-dose testing. At 10 M, 6b
exerts greater anti-
cancer activity than BMS-199264 (Figure 9), despite it having less effect on
F1 F0-ATP
synthesis, because it inhibits the reverse mode of ATP synthase more potently
than BMS-
199264. Again, a vindication that the molecules of this disclosure exert anti-
cancer activity
.. by inhibiting the reverse mode of ATP synthase, which distinguishes them
from the
macrolides and, distinctly, makes them usable therapeutically. Indeed,
molecules of this
disclosure don't appreciably inhibit the forward mode of ATP synthase, in
sharp distinction
to the macrolides. The compounds that contain a protonable nitrogen atom in
their imidazole
reduce Fi Fo-ATP synthesis in SMPs because they shuttle protons across the
mitochondrial
inner membrane, dissipating the proton motive force (uncoupling). Figure 31
presents
structure-activity data for such uncoupling in whole cells, using compounds
that are also
componentry to this invention as anti-cancer drugs. BMS-199264 (logP = 3.79,
calculated
[25]) uncouples more than 6b (logP =5.97, calculated [25]) because its logP is
closer to the
logP = 3.2 (calculated) optimum for uncoupling [36].
19a is a racemate, wherein the S stereoisomer, and not the R stereoisomer,
potently inhibits
FIFO-ATP hydrolysis [5-6]. I tried to test the anti-cancer activity of the
separated
stereoisomers. They were successfully separated by chiral supercritical fluid
chromatography
(SFC). But subsequently underwent racemization during the NCI-60 tests. One
stereoisomer
sample conveyed slightly better anti-cancer activity than the other, revealing
more area under
the curve for S stereoisomer exposure, and possibly a slight enduing
enantiomeric excess (ee)
of S stereoisomer. Both samples ultimately contained a significant proportion
of S
stereoisomer and both had strong anti-cancer activity (Figure 8). The Pearson
correlation
coefficient (R = 0.8 at 10 AM; R = 0.9437 at 100 M) for their patterns of
anti-cancer activity
is significant (at p < 0.00001). Racemization of the S stereoisomer is slowed
by replacing the
hydrogen atom on its chiral carbon with a deuterium atom (enrichment) and this
is a new
composition of matter, which is componentry to this invention, as is the
method/process of
using it for anti-cancer therapy. With this modification, the enantiomeric
excess (ee) of the
eutomer endures for longer and so per-unit anti-cancer activity is better, for
longer. Analogy
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by the macrolide inhibitors of [14], which [PI, P2, P3] teaches, would suggest
that the Sand
R stereoisomers have equal anti-cancer activity, and that this would be weak,
because they
are both comparably weak reducers of FiFo-ATP synthesis (EC50> 100 M in SMP
assays).
By contrast, by the invention of this disclosure, the S stereoisomer
specifically is revealed to
be a potent anti-cancer therapeutic.
STEREOISOMERISM
For some molecules of this disclosure, one of its stereoisomers has much lower
1050 than the
other for inhibiting FIFO ATP hydrolysis, and so, by the invention of this
disclosure, this is
.. the preferred stereoisomer for anti-cancer use. Indeed, a form with high
enantiomeric excess
(ee) for this preferred stereoisomer is the preferred embodiment for anti-
cancer therapy, e.g.
ee = >70%, ee = >95%, >99% more preferred, =100% most preferred. However, ee
can be
eroded by racemization. This invention discloses an improvement. Embodied by
this
disclosure are permutations of each of its chiral molecules, wherein the
hydrogen attached to
each chiral carbon is replaced with a deuterium, wherein the natural abundance
of deuterium
(0.015%) at this position is enhanced (non-limiting example: >3000 times
greater than the
natural abundance of deuterium, i.e. a >40% incorporation of deuterium). The
deuterium
Kinetic Isotope Effect (KIE) [37] slows racemization.
PREFERRED EMBODIMENTS
....-- N ....--N
----N --N
N 0 N 0
A
S N NH S N NH
H H
CI CI
1110 CI CI
CI CI
hydrogen (H) at chiral centre deuterium (D) at chiral centre
More preferred
CA 3050553 2019-07-25
The structure on the left has a low EC50 against FIR' ATP hydrolysis (0.018
M), its [EC50
F Fo ATP synthesis/EC50 F IF ATP hydrolysis] ratio > 5,556. In rats, this
drug (administered
in polyethyleneglycol:water:ethanol, 1:1:1) is orally bioavailable (47%) with
good
pharmacokinetics (intravenously applied drug half-life in blood = 2.1 hours,
Cmax = 21 M,
.. volume of distribution = 2.37 1/kg). The deuterated analogue on the right,
wherein the
hydrogen atom on the chiral carbon is replaced with deuterium, conferring
greater
stereoisomeric stability because of the kinetic isotope effect (KIE, [37]) is
more preferred.
The greater the % deuterium enrichment at the chiral carbon (carbon atom
number 21) and
the greater the enantiomeric excess, the more preferred the embodiment. In
other preferred
embodiments other atoms or isotopes or groups are in place of hydrogen on the
chiral carbon,
blocking its racemization, ensuring enduring stereoisomeric excess. For
example, a halogen
e.g. fluorine or chlorine or bromine or iodine. Or carbon (methyl). Or a
bigger group such as
CH2OH.
The most valuable innovation of this invention is not a presented structure
but a discovered,
disclosed principle: the best anti-cancer compound of this invention is a
molecule that
inhibits PIP ATP hydrolysis as potently and specifically as possible, whilst
it inhibits, by
direct binding, the forward mode of the ATP synthase molecule as little as
possible: most
preferably not at all.
DESTROYING CANCER'S IMMORTALITY, lengthening lives
Unlike normal adult cells, cancer cells are immortal e.g. Hela cancer cells
have replicated
>>billions of times in laboratories across the world after their originator,
the person they
killed, Henrietta Lacks, is sadly long since dead. Figure 18 interprets
(particularly)
experimental data of Figures 8, 15 and 17; interpretation expanded upon here.
The core:
Removing cancer's immortality, rendering it mortal, removes its danger without
hurting
normal cells, which are mortal anyhow. A hyperpolarised TN correlates with the
most
dangerous cancers [38-43]. I argue because this feature/mode is a function of
cancer
proliferation and thus, the more aggressive/dangerous the cancer, the more
time they spend in
this operating mode, and thus the more chance it is detected at scale by Tim
imaging. In these
cancer cells, Tim = around -200 rather than the -140 mV in normal adult cells.
ROS cause
transient growth arrest to permanent growth arrest, to apoptosis or to
necrosis, dependent on
the level of ROS [44]. Constitutively activating oxidative phosphorylation
(OXPHOS) in
cancer cells halts their proliferation [45-46] or pushes them into apoptosis
[45-49], via its
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inherent ROS production [47]. In cancers, a higher glycolytic rate drives
greater FIN ATP
hydrolysis, greater pmf and a hyperpolarised Tim (-200 mV). This reduces the
"sink" drive
for electrons to enter the respiratory chain (especially when there is high
lactate
dehydrogenase expression (out)competing for NADH) and so decreases OXPHOS
rate,
which decreases ROS production, whilst simultaneous aforementioned features of
FiFo ATP
hydrolysis increase [NADPH] and ROS mitigation. Thus, cancers have decreased
ROS
production and increased ROS mitigation. The outcome is that cancers have
lower [ROS]
than normal cells, which is integral to their enduring information fidelity,
which permits their
"limitless replicative potential" (immortality), which confers their danger.
ROS sensing
fluorescent probes report higher EROS] in cancer than normal cells [50], and
those of the art
typically think cancers do have higher EROS], but these probes are cationic
and accumulate at
greater concentrations in cancer cells because of their more hyperpolarised
TIM (Nernst; A60
mV hyperpolarisation accumulates di-positive probe, like nitroblue
tetrazolium, 100 times
more). Other ROS sensing cancer studies report extracellular [ROS] [51] and
some cancers
overexpress NOX enzymes [52-53] at their plasma membrane, which generates
greater
extracellular [ROS], which reduces intracellular 02 and [ROS]. High mutation
rate
inextricably equals higher mutation rate, ultimately too much information
lost, thence limited
(not limitless) proliferative capability and mortality. Cancers have high
mutational load in
legacy to a higher mutation rate that excavated their lower [ROS], lower
mutation rate,
corrupted embryonic stem cell (ES) like phenotype from the genome, and as a
legacy of
cancer cells that exited this state by DNA mutation/repair, accrued mutation
at higher rate,
and mutated back into this state given its genomic proximity. In most cases by
a different
mutation route than they exited (many DNA mutation routes to a low [ROS]
destination).
Cancer is an anti-mutator phenotype, preceded and punctuated by mutator
epochs, whilst
normal adult bodies are in a runaway damage/mutation loop, aging and
ultimately dying.
Raise the [ROS] in cancer to that of a normal adult cell and cancer danger
attenuates. Indeed,
raise [ROS] less than this: cancers have many embryonic stem (ES) cell
characteristics [54-
57], such as limitless replicative potential and hyperpolarised TIM, and ES
cells respond to
ROS damage more by apoptosis than repair [57]. In cancer cells, interventional
inhibition of
FiFo ATP hydrolysis causes TIM depolarisation, more OXPHOS, greater [ROS],
which slows
cancer cell proliferation, and at greater FiF0 ATP hydrolysis inhibition, even
more OXPHOS,
[ROS], and the cancer cell dies by apoptosis or necrosis. However, once
switched into
significant OXPHOS, further FiFo ATP hydrolysis inhibition reduces OXPHOS
rate, because
less ATP needs to be synthesised because less ATP is hydrolysed by FIFO ATP
hydrolysis,
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which reduces [ROS] and anti-cancer action. So, higher doses of an FIFo ATP
hydrolysis
inhibitor can have less anti-cancer activity, seen in experimental data
disclosed herein
(Figures 8, 15, 17). Normal cells use OXPHOS and they benefit from this
slowing of
OXPHOS, less EROS], with slower aging and increased lifespan. Thus, FIE. ATP
hydrolysis
inhibitors of this disclosure hurt cancer whilst simultaneously assisting
normal cells. They
will extend lifespan in subjects with or without cancer, especially useful for
a subject(s) with
an accelerated aging disease(s) or progeroid syndrome. They will extend
healthspan by
delaying, and reducing the incidence of, the diseases of aging (any
disease/pathology whose
incidence increases with age and/or in which elevated ROS is a contributory
factor: numerous
such diseases/pathologies known to those of the art) e.g. {non-limiting}
Alzheimer's disease,
dementia, Parkinson's disease etc.). But with a caveat. This disclosure
discloses a new
fundamental biological discovery, supported by disclosed in vivo experimental
data (Figure
23): FiFo ATP hydrolysis is not a bug but a feature, necessary for heat
production and
homeothermy. Thus, inhibiting FIF0 ATP hydrolysis in a subject reduces the
subject's
endogenous heat production, which requires substitution with exogenous heat
e.g. higher
ambient temperature for the subject e.g. by appropriate geographical
(re)location. However,
whilst this issue is very serious for smaller animals such as mice (-20 g), it
is much less so
for rats (-150 g), and even less for humans (¨ 62 kg). In a simulation
reported in [58], a 30%
reduction in metabolic rate in man (70 kg, 170 cm, in 20 C ambient
temperature), maintained
for 4 hours, only decreases core body temperature by 0.18 C. By contrast,
experimental data
in [58] shows that 30% reduction in metabolic rate significantly decreases
core body
temperature in mice, and less in rats. The flip side of this is that because
much more of a
small animal's metabolism comprises futile cycling of ATP synthesis and
hydrolysis, to
generate heat, an FIFO ATP hydrolysis inhibitor can decrease their OXPHOS rate
by a greater
percentage, which will increase their lifespan by a greater percentage, than
for a larger
animal. Indeed, FIR, ATP hydrolysis inhibitors of this invention will, if
ambient temperature
is conducive, increase the lifespan of small animals immensely and an
embodiment of this
invention is to enter a compound(s) of this invention into a competition to
extend a rodent's
lifespan, wherein there is a financial prize or other benefit e.g. the Mprize.
FIFO ATP hydrolysis inhibitors of this invention confer greater life extension
in an animal
than for normal cells in culture because in culture the decrease in electron
flow along the
respiratory chain decreases ROS production but it also decreases 02
consumption and this
raises p02, which increases ROS production, whereas in an animal, their
breathing slows to
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CA 3050553 2019-07-25
keep tissue p02 (and pCO2) constant. An invention embodiment is to use an FIR,
ATP
hydrolysis inhibitor of this invention in co-therapy with a respiratory
stimulant (non-limiting
e.g. doxapram) to elevate blood and tissue p02 to more strongly
treat/ameliorate/prevent/combat cancer in a subject. Indeed, relevantly, a
compound of this
.. invention ¨ almitrine ¨ increases blood p02 in humans.
In an invention embodiment, one or more administered FIFO ATP hydrolysis
inhibitors of this
invention, or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug
thereof, reduces
the ROS generation per unit time in a subject, which reduces their DNA
damage/aging rate,
such that it becomes lower than their DNA repair rate, and so their DNA (and
other) repair
mechanisms are under rather than overwhelmed, and their aging is stopped
(repair matches
damage rate) or reverses (greater repair than damage rate) so the subject
becomes
biologically younger, rather than older, in chronological time.
.. In an invention embodiment, one or more FIR ATP hydrolysis inhibitors of
this invention, or
a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, are
administered to a
subject topically/locally rather than systemically, optionally to a cancer(s)
or close to a
cancer(s) or to a blood vessel perfusing a cancer(s), wherein this cancer can
be a tumour, and
thence the compound(s) conferred reduction in heat generation (and slower
aging) is
.. disproportionally applied to this localized region, wherein its lesser heat
generation is offset
by heat transfer from surrounding body area(s), especially given the heat
distributing nature
of blood flow. In a particular embodiment, the cancer is suspected rather than
diagnosed. In
an embodiment, a compound(s) of this invention is applied topically to the
skin, optionally to
a skin cancer(s).
Eukaryotes must maintain a hyperpolarised Tim or they will undergo apoptosis
[59]. In
cancers that cannot use OXPHOS because of deficiency(s) in their respiratory
chain, or
because of hypoxic/anoxic environment (tumours are often hypoxic), FIF0 ATP
hydrolysis is
the only means they can maintain Tim, which an FIR ATP hydrolysis inhibitor
undermines
.. and the subsequent Tim depolarisation triggers cancer apoptosis.
TREATING CACHEXIA
Cancers can utilise aerobic glycolysis (Warburg effect) at one or more stages
of their cell
cycle. ATP yield from one glucose molecule is 2 ATP by aerobic glycolysis and
¨30 ATP by
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CA 3050553 2019-07-25
oxidative phosphorylation [1-3]. The former produces 2 molecules of lactate
for each glucose
consumed, which can be converted, at the cost of 6 ATP, to glucose by the Cori
cycle in the
liver [1]. Thence, in this case, the overall ATP yield of aerobic glycolysis
is -4. Assuming
equal energy use, and assuming the cancer is always rather than
disproportionally using
aerobic glycolysis, 1 g of cancer uses ¨34 times more (potential) energy than
1 g of normal
tissue. However, this is likely an underestimate because cancers typically
have higher energy
use: cancer uses FiFo ATP hydrolysis, with protons returning to the
mitochondrial matrix by
UCP2 (overexpressed in many cancers [60-61], to burn glycolytic ATP. This
releases
glycolytic enzymes from ATP feedback inhibition and permits high glycolytic
and PPP rate,
producing glycolytic intermediates for biosynthesis and elevated [NADPH] for
increased
ROS mitigation, thence low [ROS], releasing significant lactate. Elevated
blood [lactate]
correlates with cancer danger [27]. So, if food intake does not increase upon
cancer
developing, a growing cancer can deny energy to normal tissues, which
atrophies them,
which means even more energy is available to the cancer, which grows further
and a positive
.. feedback loop (cachexia) ensues which is the leading cause of death in
cancer patients. By
this invention, FIFO ATP hydrolysis inhibitor(s) switch cancers out of aerobic
glycolysis, into
OXPHOS, with its associated ROS, aging, mortality, and break this positive
feedback loop,
treating/ameliorating/preventing/combating cancer associated cachexia in a
subject(s). FiFo
ATP hydrolysis inhibitor(s) also assist cachexia sufferers by making normal
cells more
efficient, thence requiring less glucose, and can
treat/ameliorate/prevent/combat non-cancer
driven cachexia in a subject(s) too. Cachexia occurs in many end-stage
illnesses such as
cancer, heart failure, chronic obstructive pulmonary disease (COPD), liver
failure, kidney
failure, stroke, rheumatoid arthritis, severe burn injury and HIV/AIDS. It
also occurs in
aging.
Embodied by this invention is a therapeutically effective amount of a
compound(s) that
reduces FIR) ATP hydrolysis, for example a compound(s) of Formula (I-VI),
administered in
co-therapy with a therapeutically effective amount of a compound(s) that
inhibits UCP2,
optionally incorporated in same or different pharmaceutical composition(s), to
treat/ameliorate/prevent/combat cancer and/or cachexia in a subject. Genipin
and cisplatin are
non-limiting examples of compounds that inhibit UCP2, cisplatin acts on
additional targets
also.
CA 3050553 2019-07-25
BODY TEMPERATURE
Administered to a subject, F1 F0-ATP hydrolysis inhibitor(s) conserves ATP, so
less ATP
needs to be synthesized, therefore respiration rate slows, thence metabolic
heat production
declines and body temperature can fall towards ambient temperature (if ambient
< body
temperature). Experimental evidence is disclosed in Figure 23. So, when the
ambient
temperature isn't arduous (not requiring significant energy consuming
physiological/behavioural adaptations to maintain body temperature) and
dietary intake stays
constant, weight gain/maintenance can occur, which can assist cachexia, for
example cancer
driven cachexia. This is clinically valuable because cachexia is the leading
cause of death in
cancer patients. If the ambient temperature is sufficiently close to the
required body
temperature, then the aforementioned decrease in heat generation is safe,
because the body
temperature can't fall below the ambient temperature. So, for example, if the
ambient
temperature is 37 C, inhibiting FiFo-ATP hydrolysis could make body
temperature fall to
this ambient temperature, but not below it, and this is safe because ---37 C
body temperature
is safe. Inhibiting FiFo-ATP hydrolysis will reduce, but not abolish,
metabolic heat
production. So, body metabolism will still contribute to heating the body,
just less so, which
will shift the thermoneutral and thermal comfort zones (terms well known to
those of the art
[62], temperatures vary by species, as is well known to those of the art) to
higher
temperature(s). If the subject is located at a higher temperature to account
for this shift, for
example at their updated, higher thermoneutral temperature, or make
behavioural adaptations
(e.g. wearing more clothes), then this shift is harmless. An embodiment of
this invention is
setting the dosage of a compound(s) that inhibits FIFO ATP hydrolysis with
consideration of
the ambient temperature, wherein higher dosages are permissible at higher
ambient
temperatures. The preferred ambient temperature for a dosage permits the
subject to be
thermoneutral, and/or thermal comfortable, without the metabolic heat
(respiration) fraction
driven by the FIR) ATP hydrolysis that is lost because of this dosage. This
temperature
management issue is more important for smaller than larger animals, because
surface area
scales to mass by a fractional power (e.g. refer Kleiber's law) and so larger
animals retain
their generated heat better, and so a given percentage drop in (per unit mass)
metabolism will
cause a smaller drop in body temperature in a bigger animal. The
aforementioned weight gain
can be of great clinical/health/nutritional value, or aesthetic value (by non-
limiting example:
bodybuilders), or commercial value when applied to livestock/farm animals or
any animal
with a commercial value e.g. racing animals, such as horses. This invention
encompasses a
method/process of using a compound(s) of this disclosure for these
applications, or any others
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wherein weight, nutritional or energetic gain is wanted in an animal or human.
An
embodiment of this invention is a method in which a subject takes or is
administered an
effective amount of a compound(s) of this invention, for example a compound of
Formula (I),
(II), (III), (IV), (V) or (VII) or another compound that selectively inhibits
FiFo ATP
.. hydrolysis, or a pharmaceutically-acceptable salt, solvate, hydrate or
prodrug thereof, to
treat/ameliorate/prevent/combat cachexia, cancer-associated/driven cachexia,
weight loss or a
disease or disorder or physiological process or environmental temperature that
causes a
higher than normal body temperature (many known to those of the art, only a
subset listed
here to illustrate and not restrict the invention) which can include, but
isn't limited to,
ingesting an uncoupler (e.g. 2,4-dinitrophenol), infection, sepsis,
neutropenic sepsis, stroke,
fever, pyrexia, hyperpyrexia, hyperthermia, malignant hyperthermia,
neuroleptic malignant
syndrome, serotonin syndrome, toxic serotonin syndrome, thyroid storm,
heatstroke, surgery
related, menopause ("hot flushes"), infection (non-limiting e.g. roseola,
measles, enteroviral
infections, parasitic, viral, fungal, Chlamydial, Rickettsia], bacterial,
mycobacterial, systemic
bacterial, intravascular, HIV associated, nosocomial), pyrogenic infection,
thermoregulatory
disorder(s), connective tissue disease(s), Kawasaki syndrome, drug overdose,
drug induced
hyperthermia, alcohol/drug withdrawal, idiosyncratic drug reaction, fever of
known (non-
limiting e.g. infectious disease(s), inflammation, immunological disease(s),
non-infectious
inflammatory disease(s) {non-limiting eg. systemic rheumatic and autoimmune
diseases,
vasculitis, granulomatous diseases, autoinflammatory syndromes}, tissue
destruction,
reaction to incompatible blood product(s), metabolic disorder(s), inherited
metabolic
disorder(s), cancer, neoplasm, endogenous or exogenous pyrogen(s), injury,
head injury) or
unknown or uncertain origin, or to cause greater metabolic/bioenergetic
efficiency in the
subject, enhancing their physical and/or mental performance and/or causing
body weight
gain, and/or to confer hypothermia in a subject for some medical or other
purpose which can
include, but isn't limited to, any disease/disorder/pathology/injury/surgery
that hypothermia
has been/is shown to benefit, slowing a chemical reaction(s) rate in a subject
for therapeutic
benefit, preventing/minimizing brain and/or tissue damage, slowing
physiological/pathological processes (reaction rates are temperature
dependent) and so
"buying time" to get the subject to treatment for their emergency (e.g.
trauma/septic shock or
other medical emergency), slowing the progress of sepsis until a sufficient
concentration of a
working antibiotic(s) can be built up in the subject (furthermore hypothermia,
by slowing
sepsis progression, buys time to observe which antibiotic(s) can work,
yielding time to try
alternative further antibiotic option(s) if required), used soon after or just
before clinical/legal
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CA 3050553 2019-07-25
death to preserve the subject's organs/tissues until the subject can be
frozen/cryogenically
frozen or the pathology that caused clinical/legal death (e.g. wound) can be
fixed and the
subject resuscitated, administered to a subject when a first responder (e.g.
ambulance crew)
deems the subject dead or unlikely to survive the journey to a medical
facility (e.g. hospital)
wherein this administration helps to preserve the subject which is helpful if
hospital staff
subsequently assess that they can, or might be able to, save the subject,
stabilizing
surgical/trauma/Emergency Room (ER) patients, deep hypothermic circulatory
arrest for
surgery (DHCA, non-limiting applications of DHCA include repairs of the aortic
arch, repairs
to head and neck great vessels, repair of large cerebral aneurysms, repair of
cerebral
arteriovenous malformations, pulmonary thromboendarterectomy, resection of
tumors that
have invaded the vena cava, brain tumor resection {wherein the anti-cancer
activity of a
compound(s) of this invention juxtaposes well)), Emergency Preservation and
Resuscitation
(EPR), hypothermia for a surgical purpose, protective hypothermia during
surgery and/or
surgery complication, hypothermia to slow/reduce blood loss, hypothermia for
neuro- and/or
.. cardio- and/or organ/tissue and/or life protection in a subject that has
trauma/brain
trauma/polytrauma/surgery/stroke/ischemic stroke/hemorrhagic stroke/cardiac
arrest/myocardial infarction/hypoxia/shock (including, without limitation, low
volume,
cardiogenic, obstructive, and distributive shock)/sepsis/septic shock/multiple
organ
dysfunction syndrome/systemic inflammatory response syndrome (SIRS)/organ
.. failure/cytokine storm/anaphylactic shock/seizure/disseminated
intravascular
coagulation/blocked
airway/croup/rhabdomyolysis/[head/facial/spinal/chest/abdominal/ballistic/knife
injury/trauma], or some other medical
emergency/condition/disorder/disease/injury/operation,
hypothermia for cardiac and/or cardiovascular surgery and/or open heart
surgery and/or brain
surgery (neurosurgery) and/or surgery using total circulatory arrest and/or
surgery using
cardiopulmonary bypass, Emergency Preservation and Resuscitation (EPR),
preserving
detached body parts such as limbs and/or organs (for example during organ
storage/transport
and/or transplant, thus increasing the time window for transplantation of
organs to recipients;
compound(s) of this invention is administered to organ to be transplanted [by
administration
to donor and/or by administration to isolated organ] and/or to organ
recipient, optionally
during transplant operation), protective hypothermia, targeted temperature
management,
therapeutic hypothermia, hypothermia therapy for neonatal encephalopathy,
neonatal
hypoxia-ischemia, birth asphyxia, hypoxic-ischemic encephalopathy (HIE),
haemorrhage,
hypovolemia, exsanguination, suspended animation, decompression sickness, burn
injury(s)
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including skin burn, inflammation, allergic reaction, anaphylaxis,
tissue/organ rejection,
hypoxia, hypoxemia, anoxemia, anoxia, anemia, hypervolemia, altitude sickness,
obstructed
airway, asthma attack, hypoxia in a body/tissue/organ, hypoglycemia,
reperfusion injury
(ischemia-reperfusion injury), upon release of a ligature or tourniquet,
uraemia, crush
syndrome, compartment syndrome, traumatic brain and/or spinal cord injury,
major trauma,
infection, bacterial and/or viral infection(s) (non-limiting e.g. meningitis),
sepsis, septic
shock, stroke, cerebrovascular disease, ischemic brain injury, ischemic
stroke, traumatic
injury, brain injury, spinal cord injury, cardiac arrest, heart failure,
congestive heart failure,
Dilated cardiomyopathy, valvular heart disease, pulmonary embolism, adrenal
crisis,
Addisonian crisis, hypertensive emergency, haemorrhagic (hypovolemic) shock,
cardiogenic
shock, neurogenic shock, hepatic encephalopathy, blood loss, ischemic
brain/heart/kidney/intestinal injury, neuroprotection and/or cardioprotection
and/or tissue
protection during/after a stroke and/or ischemia and/or cardiac arrest and/or
resuscitation
and/or a period(s) of poor blood flow anywhere in a subject, or to confer
hypothermia to
treat/ameliorate/prevent/combat a poisoning by a toxic amount of a compound(s)
in a subject
(non-limiting e.g. carbon monoxide/methanol/heavy metal/ethylene
glycol/pesticide
poisoning, snake/spider/bee/insect/lizard venom, metabolic poison, nerve
agent, chemical
weapon, bacterial toxin(s) {food poisoning, Salmonella poisoning},
endotoxemia, eukaryote
produced toxin(s) e.g. (non-limiting) brevetoxin, drug/substance overdose e.g.
(non-limiting)
.. heroin, ethanol, a prescription medication(s), an over the counter
medication such as aspirin,
paracetamol etc.; hypothermia is protective to toxic insult), or to confer one
or more of
sedation, anaesthesia, hypoactivity, hibernation, torpor, suspended animation,
life extension
in a subject. In an embodiment, the amplitude of hypothermia is controlled by
setting the
ambient temperature, wherein an effective amount of administered FiFo ATP
hydrolysis
inhibitor reduces subject body temperature to the ambient temperature, and so
hypothermic
amplitude is controlled by controlling ambient temperature. Another embodiment
is that the
body temperature that the body falls to, upon administration of an effective
amount of FiF0
ATP hydrolysis inhibitor, is controlled by controlling feature(s) of
electromagnetic radiation
upon the subject, for example emergent from a radiation heater(s), optionally
controlled by
servocontrol, with the set point set at the desired hypothermic body
temperature, used as a
body heating system alone or in combination with other body warming devices
and methods
(many possibilites known to those of the art), which are optionally controlled
by
servocontrol, optionally integrated into the same control loop, optionally
used by themselves
alone or in combination for this body heating purporse, to "catch" and offset
the hypothermic
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CA 3050553 2019-07-25
drive, of an effective amount of FIFO ATP hydrolysis inhibitor(s) in the body,
at some desired
hypothermic body temperature.
Many cancers cause fever in a subject. By the invention of this disclosure,
FIR ATP
.. hydrolysis inhibitors exert anti-cancer activity and can reduce body
temperature if [ambient <
.body temperature]. Thence an embodiment of this invention is a method in
which a subject
takes or is administered an effective amount of a compound(s) of this
invention, for example
a compound of Formula (I), (II), (III), (IV), (V) or (VII) or another compound
that selectively
inhibits FIE. ATP hydrolysis, or a pharmaceutically-acceptable salt, solvate,
hydrate or
.. prodrug thereof, to treat/ameliorate/prevent/combat cancer and cancer
associcated fever,
especially (but without restriction) one or more of the following cancers,
which are well
known by those of the art to drive fever in many cases: non Hodgkin lymphoma
(NHL),
Hodgkin lymphoma, acute leukaemia, kidney cancer (renal cell cancer), liver
cancer
(hepatocellular carcinoma), bone cancer, adrenal gland tumours such as
phaeochromocytomas, tumours in the hypothalamus, solid tumours.
Compounds of this invention, which inhibit FIR) ATP hydrolysis, have utility
for making
animals and/or humans feel more comfortable in hot weather, climates and
geographies. For
example, being taken by or administered to people, especially the elderly,
during hot
summers.
This temperature aspect to compounds of this disclosure isn't relevant to the
NCI-60 tests.
Because in these studies, the ambient temperature is controlled at 37 C [35],
which is
optimal for cells, and so if these drugs make cellular temperature fall to
ambient temperature,
.. this is not detrimental. It can be an issue for laboratory animal studies
though. Laboratory
mice, for example, are typically kept at room temperature (e.g. 20 to 23 C)
which renders
them very reliant upon additional metabolic/physiological/behavioural heat
production
because their thermoneutral zone is much higher, at 30 to 32 C (can vary
depending on
strain, size, age, gender etc. [62]). An administered compound(s) of this
disclosure, which
inhibits FIFO ATP hydrolysis, can add to the cold stress that laboratory mice
endure when
kept at typical room temperature. An embodiment of this invention is the
process/method of
keeping laboratory animals at, or close to, their thermoneutral zone when
performing animal
studies with a compound(s) of this disclosure. For example, keeping mice at 30
to 32 C. And
in a further embodiment, at even higher temperature to compensate for the
amount that an
CA 3050553 2019-07-25
administered compound(s) of this disclosure, by inhibiting FIB) ATP
hydrolysis, shifts the
animal's thermoneutral zone to a zone of higher temperature. The amount
shifted will depend
on the administered dosage, so in a further embodiment, the ambient
temperature is set
according to the dosage used. Wherein, for a compound of this disclosure, a
higher ambient
temperature, within safe limits, can make a greater compound dosage safer.
An embodiment of this invention is a method in which a subject takes or is
administered an
effective amount of a compound(s) of this invention, for example a compound of
Formula (I),
(II), (III), (IV), (V) or (VII) or another compound that selectively inhibits
FiFo ATP
hydrolysis, or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug
thereof, to
treat/ameliorate/prevent/combat a medical disease/disorder, wherein the
subject is monitored,
for example by a healthcare/research professional/worker (doctor, oncologist,
nurse, vet,
pharmacist, laboratory technician, scientist) or machine/artificial
intelligence substitute, for
any adverse signs/symptoms/non-normality after compound administration (in an
embodiment for 5 minutes, in a further embodiment for 10 minutes, and in a
further
embodiment for longer) and in a particular embodiment for signs of reduction
in body
temperature (methods well known to those of the art, in a particular
embodiment the subject's
body temperature is monitored) and/or the dosage administered is set, and/or
modified (e.g.
increased in graduations), by information from this subject
wellness/normality/temperature
monitoring and/or the subject is located at an ambient temperature (e.g. in a
temperature
controlled room/enclosure/confine/climate and/or their body temperature is
modified/regulated/interdicted by heating effect of electromagnetic radiation
e.g. infrared)
that maintains their body temperature within safe limits whilst they have an
effective amount
of compound in their system. An embodiment of this invention is the
process/method of
considering the ambient temperature in the decision of whether to take or
administer a
compound(s) of this disclosure, and at what dosage. In an embodiment, a period
of medical
observation, by a clinical or healthcare professional (e.g. pharmacist),
occurs after the subject
takes or is administered a compound(s) of this disclosure for the first time,
and in a further
embodiment when the compound dosage is increased or decreased. In a further
embodiment,
during this period of medical observation, the subject stays in a location
that has medical
facilities and/or expertise to treat/combat hypothermia (well known to those
of the art), in
non-limiting example embodiments this is a hospital or clinic or pharmacy or
workplace of
healthcare professionals. In an embodiment, during this period of medical
observation, the
patient stays in a temperature controlled room or area, or at a location where
one is available
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CA 3050553 2019-07-25
nearby, and if the patient displays signs or symptoms of hypothermia, feels
uncomfortable, or
their body temperature falls, they can be located in a higher ambient
temperature. In an
embodiment, while the subject takes or is administered a compound(s) of this
invention, or in
a monitoring period after it, they stay in a room/confinement/location at a
safe ambient
temperature for having a compound(s) of this disclosure (non-limiting
examples: wherein the
ambient temperature is close to the desired body temperature, ¨37 C, or
exceeding it within
safe limits) and are monitored by observation, and in a further embodiment
their body
temperature is monitored (methods well known to those of the art), as the
controlled
room/confinement/location temperature is reduced to a different temperature,
in a further
embodiment to, at or near, the ambient climatic temperature of that geography
at that time, or
colder. In a further embodiment, this process/method is iterated until the
greatest dosage is
found at which the subject has a safe body temperature at, or near, the
ambient climatic
temperature of that geography at that time or at the ambient temperature(s) at
which the
subject will spend their time at over their course of compound administration,
or that their
ambient temperature might fall to at some time over their course of compound
administration,
wherein the course of compound administration is the period during which the
subject has an
effective amount of compound in their body.
Many clinical oncology centres have equipment for body heating, for
administrating
hyperthermia, for anti-cancer treatment. And other clinical specialities have
body heating
equipment for heat therapy. An invention embodiment is to administer an
effective amount of
an FIFO ATP hydrolysis inhibitor(s) to a subject, who is heated by such body
heating
equipment/apparatus, or other body heating equipment/apparatus. An embodiment
of this
invention is a method in which a subject takes or is administered an effective
amount of a
compound(s) of this invention, for example a compound of Formula (I), (II),
(III), (IV), (V)
or (VII) or another compound that selectively inhibits FIFO ATP hydrolysis, or
a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, to
treat/ameliorate/prevent/combat a medical disease/disorder, wherein the
subject is warmed by
a device(s), for example a medical device(s) (numerous examples known to those
of the art
e.g. refer [63-64]), to maintain their body temperature within a safe range,
optionally locating
in an incubator and/or in/under a radiant heater, optionally a scale up of
that often used for
(especially premature and/or low birth weight) babies (such scale ups, e.g.
adult radiant
warmer(s), are commercially available, used for example when patients undergo
general
anaesthesia, which can reduce metabolic rate by 20-30% [63]; teaching of [64]
incorporated
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CA 3050553 2019-07-25
herein by reference, as applied to adult or infant), and the subject's body
temperature is
maintained by heating the air to a desired temperature and/or by controlled
electromagnetic
radiation (e.g. infrared, preferably IR-A, from 0.78 to 1.4 gm), and/or by
servo-controlling
the body temperature at a desired set-point, most preferably at a body
temperature conducive
to keeping the subject alive e.g. at or near 37 C and/or at a thermoneutral
temperature for the
subject (in a further embodiment the desired set-point is set at a lower
temperature than a
normal body temperature, to induce hypothermia in the subject for a medical
purpose).
Servocontrol in this context, as well known to those of the art, refers to an
electronic
feedback system which maintains a constant temperature at the site of a
thermistor (or other)
.. probe (for non-limiting example, on the skin over the abdomen) by
regulating the heat output
of an incubator and/or radiant warmer and/or other body heating device.
Componentry to this
invention is a servocontrol variant(s) whereby body temperature is recorded by
infra-red
(and/or other electromagnetic) emission from the body and/or body temperature
is increased
by infra-red (and/or other electromagnetic) emission towards the body,
optionally where
.. temperature detection and heating occurs at different wavelengths,
optionally for use for
heating a subject with an effective amount of a compound(s) that inhibits FIFO
ATP
hydrolysis in their body, optionally a compound(s) of Formula (I-V) or (VII).
An
embodiment of this invention is a method in which a subject is administered an
effective
amount of a compound(s) or pharmaceutical composition(s) that inhibits FIE)
ATP
hydrolysis, by intravenous injection, wherein the volume injected is heated to
be at or close to
normal body temperature and/or wherein any method(s), including equipment,
used to
treat/ameliorate/prevent/combat anaesthetic associated hypothermia (methods
are well known
to those of the art) is used for a subject administered with a compound(s)
that inhibits FiFo
ATP hydrolysis, optionally a compound(s) of Formula (I), (II), (III), (IV),
(V) or (VII). An
embodiment of this invention is a method in which a subject takes or is
administered an
effective amount of a compound(s) of this invention, for example a compound of
Formula (I),
(II), (III), (IV), (V) or (VII) or another compound that selectively inhibits
FIFO ATP
hydrolysis, or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug
thereof, to
treat/ameliorate/prevent/combat a medical disease/disorder, wherein the
subject wears one or
.. more clothes, optionally a hat, to maintain body temperature within
safe/comfortable limits as
the FIR ATP hydrolysis inhibitor(s) reduces metabolic heat production. An
embodiment of
this invention is a method in which a compound(s) of this invention, for
example a compound
of Formula (I), (II), (III), (IV), (V) or (VII) or another compound that
selectively inhibits FiFo
ATP hydrolysis, or a pharmaceutically-acceptable salt, solvate, hydrate or
prodrug thereof, is
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CA 3050553 2019-07-25
distributed, sold and/or administered with a verbal and/or written
communication, optionally
in a paper insert/leaflet in a packet(s) containing the compound(s)
(optionally called
"instructions for use", and/or "prescribing information" and/or "patient
information leaflet"),
that this compound(s) can reduce body temperature and, in a further
embodiment,
communicating that should the subject that has taken or been administered one
or more of
these compounds feel cold, and/or has a reduction in body temperature, they
should do one or
more of: wear more clothes, wear warmer clothes, locate in a hotter
environment, tell a doctor
or pharmacist, go to a hospital. Each of these aforementioned communications
is a separate
invention embodiment and combinations of these are further embodiments. In a
yet further
embodiment, communicating that this is a more serious problem in children,
optionally
communicating that this is because children have a larger surface area to
volume ratio than
adults, optionally communicating that this problem is especially acute with
babies and
optionally communicating that this compound(s) should not be administered to
babies (in a
further embodiment, unless the baby is in a controlled temperature environment
such as an
infant incubator or radiant warmer). In another embodiment, a method in which
a
compound(s) of this invention, for example a compound of Formula (I), (II),
(III), (IV), (V)
or (VII) or another compound that selectively inhibits FIR) ATP hydrolysis, or
a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, is
distributed, sold
and/or administered with a verbal and/or written communication, optionally in
a paper
insert/leaflet in a packet(s) containing the compound(s) (optionally called
"instructions for
use", and/or "prescribing information" and/or "patient information leaflet"),
that alcohol
shouldn't be consumed in large amounts, and in another embodiment not at all,
if the subject
is taking or being administered such a compound(s). Optionally communicating
because
alcohol can corrupt thermoregulation, which can potentially, negatively
interact with the
thermoregulatory effect(s) of the administered compound(s). Optionally
communicating the
same caveat/warning/communication in reference to other drug(s) that can
disrupt
thermoregulation, many of which are known to those of the art (non-limiting
examples:
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CA 3050553 2019-07-25
phenothiazines {like chlorpromazine etc.), thioxanthenes etc.), in place of or
in addition to
the communication relating to alcohol.
BODY HEAT BALANCE EQUATION (1st Law of Thermodynamics)
S = M-W-E-C-K-R;
S = storage of heat in body (=0=heat balance, i.e. no change in body
temperature,
positive=increased body temperature, negative=decreased body temperature);
M = metabolic heat production (always positive in living organisms);
W = work (positive = useful work accomplished, negative = mechanical work
absorbed by
body);
E = evaporative heat transfer (positive = transfer to environment);
,
C = convective heat transfer (positive = transfer to environment);
K = conductive heat transfer (positive = transfer to environment);
R = radiant heat exchange (positive = transfer to environment);
This concept is well known ([651, herein incorporated in entirety) and
numerous strategies to
keep S=0, as M decreases when an FIR ATP hydrolysis inhibitor is administered
to the
subject, will be apparent to those of the art. For (non-limiting) example, if
M decreases, use
an infra-red lamp to make R negative and substantial enough to offset the
decrease in M
(thence keep S=0), or increase the air temperature to make C negative and
substantially
negative to offset the decrease in M (thence keep S=0), or wear clothes to
make positive
values of one or more of E, C, K, R less positive to offset the decrease in M
(thence keep
S=0).
TRANSLATING DOSAGE BETWEEN SPECIES
Larger species have lower mass specific basal metabolic rate, because this
parameter scales to
animal mass by a negative fractional power (e.g. refer Kleiber's law). Thence
larger species
metabolise drugs slower and so require, and can withstand, a lower mg/kg drug
dosage, and
this is factored into converting a drug dosage in one species (e.g. mouse)
into one of different
size (e.g. human), as is well known to those of the art (e.g. refer [66] and
the FDA guidelines
it refers to). However, by the invention of this disclosure, toxicity of
selective FIFO ATP
hydrolysis inhibitors scales to animal mass by a negative fractional power and
is less in larger
animals than predicted by conventional allometric scaling methods used in the
art. Because
larger animals have a smaller surface area to mass ratio (animal mass is
proportional to
CA 3050553 2019-07-25
animal radius3, animal surface area is proportional to animal radius2, thence
bigger animals
have smaller surface area to mass ratio), they retain metabolically generated
heat better and
thence a given percentage drop in metabolic rate doesn't cause as big a
percentage drop in
body temperature i.e. they aren't as negatively affected by a drop in
metabolic heat
production, which FIN ATP hydrolysis inhibitors cause, which is the defining
limit upon
their maximally tolerated dosage (MTD), in smaller animals at least, dambient
temperature
is significantly below optimal body temperature e.g. when ambient temperature
= ¨22 C.
Mass specific metabolic rate (MR) [62] is, assuming core body temperature
stays constant
(heat production=heat loss), and that ambient temperature is below the lower
critical
temperature, which is the lower bound of ambient temperature that an animal is
thermoneutral:
MR = C(Tb-Ta);
Tb = body temperature;
Ta = ambient temperature;
C= whole body thermal conductance= heat transfer rate (by all heat transfer
mechanisms)
from body core to environment for temperature difference of 1 C;
1=insulation=1/C;
MR = 21.66*M" 25;
C = 4.23*M-0426;
where M = body weight (g);
MR and C decrease with increasing M; MR decrease increases drug toxicity; C
decrease
decreases drug toxicity (for an FIE) ATP hydrolysis inhibitor), let us define
a danger quotient
for an Fi Fo ATP hydrolysis inhibitor: its danger is proportional to C and
inversely
proportional to MR: danger = C/MR = (4.23*M-0 426)/(21.66*M- 25). Using M=20 g
for a
mouse, and M=62 kg for a human, danger quotient for mouse (=0.12) and human
(=0.03). So,
assuming same ambient temperature for both species, which is below the
thermoneutral
temperature zone of both species, the mg/kg MTD will be ¨4 times more in
humans than in
mice, and even greater than this for a human wearing clothes (decreases C).
This is far
removed from what one of the art would expect: expecting mg/kg MTD in humans
to be kss
than in mice: for example, calculating (21.66*20025)/(21.66*62000" 25) =
20'25/62000" 25 =
¨7.5 times less, or using the more conservative exponent of [66] (and possibly
less
appropriate, at least for chemotherapeutics: refer FDA guidance cited in [663)
they would
calculate 20033/62000033 = 14.2 times less. Any given mg/kg dose in humans
will be
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metabolised (20-025)462000325) = ¨7.5 times slower and so because we have
ascertained that
the maximum permissible mg/kg dose in humans is ¨4 times more than in mice:
then the
potential anti-cancer activity of an FiFo ATP hydrolysis inhibitor is 4*7.5=-
30 times higher
in a human than a mouse, at typical room temperature. So, elucidated by the
invention of this
disclosure, small animal studies at room temperature (-22 C) will dramatically
underestimate
the anti-cancer activity possible at room temperature in humans, and severely
curtail the
therapeutic benefit fconventional allometric scaling is used to set the human
dose from mice
studies. This invention supplies a new method to better translate the dosage
of an FIFo ATP
hydrolysis inhibitor between species. However, more conventional allometric
scaling
methods can be used [66] ifthe smaller species studies are conducted at an
ambient
temperature close to optimal homeothermic body temperature, Ta;---Tb, and
especially if
humans will live at this same temperature. Thus, the temperature at which the
smaller species
study is conducted, and the temperature that the human subjects will reside
at, determines the
allometric scaling method applied, to find the equivalent human dose, which
would not have
been apparent to someone of the art without this disclosure. Furthermore, a
method as
disclosed herein can be used to modulate FIFO ATP hydrolysis inhibitor dosage
between
individuals of a single species, e.g. humans, of different sizes.
The equations and parameters used in this section illustrate the principles of
the invention and
are not restrictive. For example, other allometric exponents are contemplated
and
componentry to this invention. A method of this invention is to compare the
MTD (or some
other safety metric e.g. NOAEL, LD50, LD33 etc.) of an FiFo ATP hydrolysis
inhibitor(s)
between animal individuals and/or species of different size (e.g. mice, rats,
guinea pigs,
rabbits, dogs, primates etc.) to calculate more precisely what the particular
allometric scaling
relation is, and optionally investigate how this scaling changes with ambient
temperature.
And then use this relation(s) to calculate/triangulate/estimate a safe
starting dose in humans,
from prior animal studies. Optionally factoring in what the ambient
temperature(s) will be for
the humans administered the FiFo ATP hydrolysis inhibitor(s).
ANTI-CANCER DOSAGE
Methods to gauge the anti-cancer activity of a treatment(s) in a subject(s),
and/or to gauge the
change in anti-cancer activity associated with a change (e.g. dosage change)
in treatment(s),
are well known to those of the art. They are routinely utilised in pre-
clinical studies and
clinical practice.
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The following methods are very atypical from present dosage methods in the art
of
chemotherapy, wherein dosage is typically, simply just the maximum dosage that
the patient
can tolerate. The method of decreasing the administered dose of a compound(s)
that reduces
F1Fo ATP hydrolysis, for (non-limiting) example a compound of Formula (I) or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, in
order for it to exert
greater anti-cancer activity in a subject is an embodiment of this invention.
The anti-cancer activity of a compound(s) of Formula (I) can increase or
decrease in
proportion to an increase in compound dosage, wherein there is an optimal anti-
cancer dosage
which is not, as one of the art would expect, at the maximally tolerated dose.
Decreasing or
increasing (!) the dose away from this optimal dosage decreases anti-cancer
activity. Herein
is a method to find this optimal dosage in a subject, which is componentry to
this invention,
as are other methods to find the optimal dosage for a compound(s) of Formula
(I) or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, which
leverage/utilise
the extremely atypical dose vs. anti-cancer activity profile, disclosed herein
(Figures 8, 15
and 17).
If increased dose = increased anti-cancer activity, increase dose further. If
increased dose =
decreased anti-cancer activity, decrease dose by more than it was increased
previously. Make
the next increase in dose smaller than prior. Repeat. As this loop is
repeated, and the
increment increase in dose gets smaller, the person of the art knows that they
are increasingly
converging upon the optimal dosage. They can choose to exit the loop at any
dosage they
wish, knowing that the size of the dosage increase increment on loop exit is
an indicator of
how close they are to the optimal dosage. The smaller the dosage increase
increment at loop
exit, the closer that the dosage is to the optimum. The person of the art may
choose to do the
mirror image of the aforementioned loop and choose decreased, instead of
increased, dose
increments. There are permutations to these schemes as will be clear to
someone of the art,
now that this logic has been set out, which are componentry to this invention.
To help
illustrate (not restrict) the invention the following R [67] programming code
is disclosed,
wherein the "optimal" parameter symbolises the optimal anti-cancer dosage, and
the loop
iterates until the administered drug dosage, "dose", equals the optimal.
Someone of the art
can adjust this code, for example changing parameters and/or mathematical
symbols, to
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CA 3050553 2019-07-25
explore and understand the invention further. Of course, in reality, the
optimal anti-cancer
dosage is unknown and is not user specified, as in the code, but the code
illustrates a method
to find the optimal anti-cancer dosage for a compound of Formula (I), a method
componentry
to this invention. All parameters are merely illustrative.
# R programming code
dose = 1 # first dose to be tried
reset = dose
dt = 600 # step size that dose will be adjusted
res = dt/100000000
# res = an accuracy parameter, larger denominator means more iterations and
more accuracy
x = 0
optimum = 60
# optimal anti-cancer dose, unknown in real case, following the logic herein
finds it
n = 10
# n = an accuracy parameter, MUST be >1, larger means more iterations and more
accuracy
m = n
count = 0 # how many iterations are taken to converge
while (dt > res)
1
dose = dose + dt
if (dose<optimum)
# Increased dose = Increased anti-cancer activity, so Increase dose further
dose = dose+dt
x=x+1
1
if (dose>optimum) # Increased dose = Decreased anti-cancer activity, so
Decrease dose
if (dose<0){
dose=reset
dt = dt/n
}
dose = dose-(dt+(dt/n))
x=x-1
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CA 3050553 2019-07-25
1
if (x==0){dt=dt/m }
# if dose is changed in one direction (increase or decrease) and then
subsequently changed in
# opposite direction, decrease step size
if (abs(x)==2){x=0}
count = count + 1
# if (abs(dose-optimum)<1){break} # not used, but here for interest
}
In place of, or after, the aforementioned, or other, method has converged the
administered
dose to be close to optimum, the dosage can be set randomly, optionally
restricted within a
range, by some formal/informal random number generator. It need not be
perfectly random
and can just be the arbitrary choice of a person(s). This formalised/informal
random walk is
used to find a compound(s) dosage, of Formula (I), with greater anti-cancer
activity. It is
distinct from conventional methods in its use of dosage reduction (not merely
increase) to
seek greater anti-cancer activity.
The most optimal dosage, or dosage range, for a compound of Formula (I) is
that which
causes cancer cell death. This is a narrow dosage range (Figure 18). Flanked
at lower and
higher doses by dosages that exert less anti-cancer activity. An embodiment of
this invention
is changing the administered dosage of a compound of Formula (I) in a subject
until a
dosage(s) causes cancer cell death, optionally by apoptosis. Detected, for non-
limiting
example, by cancer regression rather than merely slowing of cancer growth
and/or by
apoptosis markers, well known to those of the art (e.g. refer [68]), for
example, in the blood.
Using a dosage selection method disclosed herein to find a good anti-cancer
dosage for a
compound of Formula (I), optionally in a xenograft/syngeneic rodent(s),
optionally housing
the rodent(s) at an ambient temperature >26 C (in further embodiments: >28,
>30, >32, >34,
.?.36 C), is componentry to this invention. As is any housing/rearing of a
rodent(s) at an
ambient temperature >26 C (in further embodiments: >28, >30, >32, >34, >36 C)
whilst
administered with a compound(s) of Formula (I), optionally to select the
starting dosage(s)
for human clinical trial(s).
CA 3050553 2019-07-25
A method of decreasing the administered dose of almitrine or a
pharmaceutically-acceptable
salt, solvate, hydrate or prodrug thereof, optionally almitrine dimesylate, in
order for it to
exert greater anti-cancer activity in a subject is an embodiment of this
invention.
Componentry to this invention is to use one or more of the aforementioned
methods,
disclosed in this disclosure section for a compound(s) of Formula (I), to find
the optimal
dosage of a compound(s) of Formula (VI) or a pharmaceutically-acceptable salt,
solvate,
hydrate or prodrug thereof, optionally almitrine dimesylate, which
leverages/utilises its
extremely atypical dose vs. anti-cancer activity profile, disclosed herein
(Figures 7 and 21).
UNCOUPLING CANCER
The meaning and characteristics of an uncoupler are well known to those of the
art e.g. refer
[36]. An uncoupler is a molecule that can bind a proton(s) in the
mitochondrial
intermembrane space (IMS), move across the mitochondrial inner membrane, and
release the
proton(s) in the mitochondrial matrix, which dissipates the proton motive
force (pmf), and
that can then return to the IMS, and repeat this sequence iteratively. 2,4
dinitrophenol is an
example of an uncoupler. Many other uncouplers are known to those of the art.
An invention
embodiment is to administrate a therapeutically effective amount of a
compound(s) that
inhibits FiFo ATP hydrolysis (e.g. a compound(s) of Formula (I), (II), (III),
(IV), (V) or
(VII)), with a therapeutically effective amount of the same or a different
compound(s) that
uncouples the proton motive force, for use in a method of treatment of the
human or animal
body by therapy, wherein optionally the FiFo ATP hydrolysis inhibitor(s) and
uncoupler(s)
are in a single pharmaceutical composition and/or are packaged, and/or
distributed, and/or
sold together, optionally for the treatment/amelioration/prevention/combat of
cancer in a
subject(s). As aforementioned, the dosage range that an F1 F0 ATP hydrolysis
inhibitor of
Formula (I) kills cancer cells is extremely narrow. This range is broadened by
co-
administration of an uncoupler(s), which increases OXPHOS rate and [ROS], and
reduces the
concentration of FIFO ATP hydrolysis inhibitor required. In some embodiments,
the anti-
cancer activity of the uncoupler(s) and FiFo ATP hydrolysis inhibitor(s)
synergize
(potentiate). Optionally, an FIR ATP hydrolysis inhibitor(s) is administered
with an
aforementioned method(s) to (fractionally) optimise its dosage, before the
administration of
the uncoupler, which then decreases the dosage of uncoupler required. An FiFo
ATP
hydrolysis inhibitor(s) decreases metabolic inefficiency and body temperature,
an
uncoupler(s) increases metabolic inefficiency and body temperature: co-
administration,
especially optimised co-administration, optionally wherein an optimised amount
of each is in
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CA 3050553 2019-07-25
a single pharmaceutical composition, can reduce the change in body temperature
that each
would cause alone; and there is synergy in anti-cancer activity yielded.
Componentry to this
invention are compounds that inhibit FiFo ATP hydrolysis and that uncouple the
proton
motive force (pmf) e.g. (non-limiting) BMS-199264. An embodiment of this
invention is to
administrate, or for the subject to self-administer, a therapeutically
effective amount of a
compound(s) that inhibits FiFo ATP hydrolysis and that uncouples the proton
motive force,
for use in a method of treatment of the human or animal body by therapy,
optionally for the
treatment/amelioration/prevention/combat of cancer in a subject(s).
In cancers that cannot use OXPHOS because of deficiency(s) in their
respiratory chain, or
because of hypoxic environment (tumours are often hypoxic), which thence
singly rely upon
FiFo ATP hydrolysis to maintain TN, an uncoupler(s) will erode their Tim and
an FIFO ATP
hydrolysis inhibitor(s) will block their only means to counter this, their TN
will collapse,
triggering their apoptosis. Given the severity of this vulnerability, and the
potentiating effect
.. of the two drugs, low drug doses apply. Meanwhile, normal cells will
maintain TN by greater
OXPHOS rate.
Also componentry to this invention is the use of a therapeutic amount of
uncoupler(s) to
treat/ameliorate/prevent/combat cancer in a subject. If TIM depolarises,
apoptosis ensues [59].
.. Aerobic respiration, favoured by normal adult cells, hyperpolarises TIM as
it produces ATP.
Aerobic glycolysis, favoured by many cancers some or all of the time, consumes
ATP to
hypeipolarise 'Pm Under the challenge of an uncoupler compound(s), the former
is more
sustainable than the latter, even more so because of the difference in ATP
yields (-30 vs. 2
ATP per glucose), and thence there is a therapeutic margin. In response, some
cancers will be
able to switch out of aerobic glycolysis, and into aerobic respiration, but
its oxidative
phosphorylation (OXPHOS) component will increase ROS, bring aging, mortality
and
mitigated danger, especially because the uncoupler(s) will drive higher OXPHOS
rate. Unlike
other chemotherapeutics, uncouplers only interact with protons, not DNA
encoded proteins,
and so their therapy cannot be resisted by DNA mutation rendered changes in
protein
structure, which is the basis to present day cancer drug resistance, which
kills.
UNCOUPLING IS VIRTUOUS
The imidazole containing compounds of this disclosure inhibit FIFO ATP
hydrolysis and
uncouple (shuttle protons across the mitochondrial inner membrane (IM),
eroding the proton
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CA 3050553 2019-07-25
motive force, pmf). The former can exert a specific anti-cancer activity,
because it
undermines the means some cancers maintain Tim in normoxia (experimentally
shown by
data of this disclosure) or in hypoxic tumours, and the compound's uncoupling
can also exert
specific anti-cancer activity, explained now. The imidazole containing
compounds of this
disclosure bind ATP synthase at or near the IFI binding site. In normal cells
they bind ATP
synthase at this site and are sequestered from uncoupling, and the ATP they
"save" by
binding and inhibiting FIR ATP hydrolysis can (over)compensate for the ATP
"lost" to their
uncoupling. But some cancers have very high IF' expression (numerous studies
show this,
e.g. refer [23]). And for some cancers, this is to inhibit FIR ATP hydrolysis,
to make their
OXPHOS more efficient, which allows them to maintain [ATP] at low [02], and
thence
survive using OXPHOS in hypoxia (their heat generation is less but their
temperature is
maintained by heat conduction from surrounding tissues). This high IF'
expression blocks the
binding of these compounds to their binding site on ATP synthase, so the
compounds aren't
sequestered from uncoupling, and this uncoupling increases the 02 requirement
of this cancer
which can't be met in the hypoxic microenvironment of its tumour, thence the
cancer's
intracellular [ATP] can't be maintained and its proliferation is slowed and/or
it dies. So,
herein, this invention discloses that the uncoupling aspect to the imidazole
containing
compounds of this disclosure can deliver additional, specific, anti-cancer
activity, for
example, against those cancers that don't rely upon FiFo ATP hydrolysis. This
invention
discloses the process/method of using a compound(s) that can inhibit FiFo ATP
hydrolysis,
and that can shuttle protons across the IM to dissipate the pmf (uncouple), as
an anti-cancer
therapeutic. Wherein the compound inhibits FiFo ATP hydrolysis by direct
interaction with
ATP synthase, and reduces Fi Fo ATP synthesis (primarily) by uncoupling. So, a
compound
needn't necessarily have a much lower ECK' for FiFo ATP hydrolysis than FIFO
ATP
.. synthesis, in an SMP assay, to be componentry to this invention as an anti-
cancer therapeutic.
Indeed, even compounds with a lower EC50 for F1F0 ATP synthesis than FIR ATP
hydrolysis
in an SMP assay can be componentry to this invention, as anti-cancer
therapeutics, provided
they do inhibit Fi Fo ATP hydrolysis and provided their inhibition of FIR ATP
synthesis is
(primarily) because of uncoupling rather than inhibiting the forward mode of
ATP synthase.
Oligomycin, for example, does not fit these requisites. So, this invention
discloses the method
of using compounds that inhibit FiFo ATP hydrolase, that don't inhibit FIE)
ATP synthase,
and that uncouple the proton motive force, as anti-cancer therapeutics.
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DEUTERATED COMPOUNDS OF THE INVENTION
Deuterium (D or 2H) is a stable, non-radioactive isotope of hydrogen and has
an atomic
weight of 2.0144. Hydrogen naturally occurs as a mixture of the isotopes 1H
(hydrogen or
protium), D (2H or deuterium), and T (3H or tritium). Herein, all percentages
for the amount
of deuterium present are mole percentages. The natural abundance of deuterium
is 0.015%.
Thence one of the art recognizes that in all chemical structures containing an
H atom(s), this
H atom(s) actually represents a mixture of H and D in the compound, with about
0.015%
being D. Therefore compounds that have a higher level of deuterium
incorporation, i.e.
compounds enriched to have a greater D incorporation than natural abundance
(>0.015%),
should be considered unnatural and so distinct from their non-enriched
counterparts. A
compound is said to be "deuterium enriched" if it has a quantity of deuterium
that is greater
than in naturally occurring compounds, or synthetic compounds prepared from
substrates
having the naturally occurring distribution of isotopes. At a lab scale amount
(milligram or
greater) it can be difficult to achieve 100% deuteration at any one site of a
compound. Herein,
when 100% deuteration is recited or a deuterium atom is specifically shown in
a structure, it
is assumed that a small percentage of hydrogen may still be present. Deuterium-
enrichment
can be achieved either by exchanging compound protons with deuterium or by
synthesizing
the compound with deuterium enriched starting materials, which are
commercially available
or can be readily prepared by someone of the art using known methods.
Embodiments of this invention include compounds of Formula (I), (II), (III),
(IV), (V), (VI)
and (VII) with one or more of their hydrogen atoms replaced by deuterium, at a
greater
frequency than the natural abundance of deuterium (0.015%). For non-limiting
example:
>3000 times greater than the natural abundance of deuterium (i.e. a >40%
incorporation of
deuterium at a hydrogen replacement position). Additional examples of the
abundance of
deuterium at a position in, or positions of, a compound embodiment of this
invention include
40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58,
59, 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 to about 100%. In certain embodiments,
the abundance
of deuterium at a position in, or positions of, a compound embodiment of this
invention is at
least 40%. In certain other embodiments, the abundance of deuterium at a
position in, or
positions of, a compound embodiment of this invention is at least 60%. In
futher
embodiments, the abundance of deuterium is at least 75%. In yet other
embodiments, the
abundance of deuterium is at least 90%. It is to be understood that the
deuterium-enriched
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CA 3050553 2019-07-25
compounds described herein can be combined with a pharmaceutically acceptable
carrier to
form a pharmaceutical composition.
In the Description and Claims of this disclosure, when a position on a
compound structure is
designated deuterium (D), or said to have deuterium, or said to be enriched
for deuterium, it
is because the abundance of deuterium at that position is not at the natural
value (0.015%) but
greater. Typically, in excess of 40%. The phrase 'enrichment at the chiral
centre' herein, for
example for a compound of Formula (I), means that the molar amount of
deuterium at the
chiral centre, as a percentage of the total amount of all hydrogen isotopes at
the chiral centre,
is greater than 0.015%, especially greater than 1%, preferably greater than
40%, more
preferably greater than 45%, and in ascending order of preference, >52.5%
deuterium
enrichment at the chiral centre, >60% deuterium enrichment at the chiral
centre, >67.5%
deuterium enrichment at the chiral centre, >75% deuterium enrichment at the
chiral centre,
>82.5% deuterium enrichment at the chiral centre, >90% deuterium enrichment at
the chiral
centre, >95% deuterium enrichment at the chiral centre, >97% deuterium
enrichment at the
chiral centre, >99% deuterium enrichment at the chiral centre, >99.5%
deuterium enrichment
at the chiral centre, 100% deuterium enrichment at the chiral centre. Greater
% deuterium
enrichment is preferred.
Further possible isotopic variants of the structures of this invention are
further embodiments
of this invention. An invention embodiment is a compound that inhibits the
reverse mode,
more than the forward mode, of ATP synthase, which has deuterium in place of
hydrogen (at
a greater frequency than 0.015% e.g. >40%) at one or more places upon its
structure, and/or
any other isotopic substitution/enrichment (at a greater than natural
frequency e.g. 13C and/or
15N enriched; '3C enrichment {abundance > [natural abundance=1.109%] } at the
chiral
carbon of compounds of Formula (I) is especially preferred, especially when,
if applicable, its
attached hydrogen is enriched for 2H {abundance > [natural abundance=0.015%];
preferred is
when '3C enrichment at the chiral carbon >40%, optionally 2H enrichment at the
chiral
carbon >40% also}).
Molecule synthesis routes described in [5, 6, 7, 8, Pl, P2, P3, P4, P5, P6]
(including
references cited therein, where appropriate, and in their supplementary
materials, all herein
incorporated in their entirety) - for synthesizing molecules that inhibit F1F0
ATP hydrolysis
more than Fi Fo ATP synthesis - are componentry to this disclosure, as
synthesis routes for
CA 3050553 2019-07-25
synthesising anti-cancer molecules. In other embodiments of this invention,
any given
molecule synthesis route described in [5, 6, 7. 8, P1, P2, P3, P4, P5, P6, P7,
P8] is used with
starting reagents, compounds, solvents and/or intermediates that have
deuterium in place of
hydrogen at some position(s). Such compounds are commercially available (e.g.
refer C/DN
Isotopes Inc., Pointe-Claire, or CK Isotopes Ltd., Desford, UK, or Cambridge
Isotope
Laboratories, Tewksbury, MA). Or they can be created in house by invoking
standard
synthetic protocols known in the art for introducing isotopic atoms to a
chemical structure.
For (non-limiting, illustrative) example, a compound can be deprotonated by
LiHMDS in
tetrahydrofuran (THF) at -78 to -40 C for 20 minutes, followed by quenching
with
deuterium oxide (D20, "heavy water"), to obtain a deuterated compound [37].
During these
steps, a group upon which hydrogen is still desired over deuterium can be Boc
protected and
this Boc group removed subsequently using trifluoroacetic acid (TFA) treatment
at room
temperature. At the end, the level of deuterium can be checked by III NMR. The
initial
deprotonation step isn't absolutely necessary as HID exchange will occur when
a molecule is
quenched with D20, and this reaction can be catalysed, by acid, base or metal
based catalysts
such as platinum. If, after D20 quenching, the level of compound deuteration
is insufficient
(observed using 1H NMR) then the compound is quenched with D20, or some other
deuterium containing solvent, for a longer period of time. Compounds of this
disclosure can
be synthesised in D20, during one or more chemical steps, or a starting
compound,
intermediate or final molecule of this disclosure can be incubated in D20 to
produce a
deuterated version(s). So, deuterium-enriched compounds of this invention can
be prepared
by substituting a deuterium-enriched reagent or solvent for a non-isotopically
labeled reagent
or solvent in the synthetic schemes reported in [5, 6, 7, 8, P1, P2, P3, P4,
P5, P6, P7, P8].
Non-limiting example embodiments of the invention are deuterated enumerations
of Markush
Formulas (I), (II), (III), (IV), (V), (VI) and Formula (VII). These
isotopologues are
componentry to the present invention as new compositions of matter, and in non-
limiting
embodiments are used singly or in combination, optionally in co-therapy with
an FDA and/or
EMA approved medicine(s) and/or treatment(s), for example a licensed cancer
treatment, for
use in a method of treatment of the human or animal body by therapy,
optionally in a method
of treating/ameliorating/preventing/combating cancer in a subject.
SILICON ANALOGUES OF THE INVENTION
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CA 3050553 2019-07-25
Encompassed by the invention is a compound or compounds of Formula (I), (II),
(III), (IV),
(V), (VI) and/or (VII) wherein one or more carbon atoms are substituted with
silicon, and in a
further embodiment wherein this compound(s) is used, or a salt, hydrate,
solvate, prodrug or
pharmaceutical composition thereof, optionally in co-therapy with an FDA
and/or EMA
approved medicine(s) and/or treatment(s), for example a licensed cancer
treatment, optionally
in the same pharmaceutical composition, for use in a method of treatment of
the human or
animal body by therapy, optionally in a method of
treating/ameliorating/preventing/combating cancer in a subject.
METHODS TO FIND FURTHER COMPOUNDS COMPONENT TO THIS
INVENTION
A method to find a compound(s) of this invention is by screening for/seeking a
compound(s)
that preferentially inhibits the reverse mode of ATP synthase. For example, by
separately
assaying (in space and/or time) a compounds's effect upon ATP synthesis and
ATP
hydrolysis by ATP synthase (in its entirety or, less preferably, a component
part of it). Then
comparing these assay results. The greater the inhibition of reverse vs.
forward mode, the
more preferred a compound is for a use of this invention. To illustrate, by
the invention of
this disclosure, the greater a compound inhibits the reverse vs. forward mode
of ATP
synthase, the more preferred this compound is for anti-cancer and/or anti-
aging use. An
invention embodiment is the process/method of seeking a new compound(s) of
this invention
by assaying whether a candidate molecule can depolarise Tim, when Tim is
maintained by
FIFO ATP hydrolysis (e.g. when OXPHOS is blocked by a respiratory chain
inhibitor(s) or
insufficient 02), but that can't hyperpolarize Tim and/or decrease 02
consumption, when Tim
is maintained by proton pumping by complexes of the respiratory chain. Such an
assay is
.. described in [121. A further method is screening a number of compounds to
find one or more
with this activity in this assay. An invention embodiment is seeking a
compound(s) of this
invention by assaying whether a candidate molecule inhibits ATP hydrolysis
more than ATP
synthesis in submitochondrial particles (SMPs), wherein a further method is
screening a
number of compounds to find one or more with this activity in this assay. ATP
hydrolysis can
be assayed by (non-limiting example) a spectroscopic assay for NADH
fluorescence that
incubates the SMPs with pyruvate kinase and lactate dehydrogenase enzymes
(assay well-
known to those of the art). ATP synthesis can be assayed by (non-limiting
example) a
spectroscopic assay for NADPH fluorescence that incubates the SMPs with
hexokinase and
glucose-6-phosphate dehydrogenase enzymes (assay well-known to those of the
art). These
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CA 3050553 2019-07-25
assays are reported in in any one of [5, 7, 8, 11, 12, 13, 70], and/or as
referenced therein, all
of which are herein incorporated in their entirety. In these SMP assays, the
criteria for a
candidate anti-cancer compound is a low EC50 against ATP hydrolysis (thence
anti-cancer
activity) and a higher EC50 against ATP synthesis (thence safe for normal
cells). These SMP
assays deliver high signal-to-noise because non-specific protein inhibiting
compounds (Pan-
assay interference compounds, PAINS), which are the bane of drug discovery
screening
assays, inhibit both ATP hydrolysis and synthesis, and thus are dismissed by
the screening
algorithm. So, the screening assay inherently screens out PAINS. This is
distinctive and
valuable.
Componentry to this invention is screening, using one or more screening assays
herein
described, compound(s) from one or more compound collections/libraries known
to, or
findable by, one of the art, optionally a proprietary compound collection(s)
{optionally a
collection(s) or sub-collection(s) that belongs to, or is sourced from, a
major/multinational
pharmaceutical company and/or a pharmaceutical company with >$50 million in
annual sales
and/or a Contract Research Organisation [CRO, illustrative example would be
Charles River
Laboratories]) and/or a publically/commercially available compound
collection(s) {or a
fraction thereof}, for example, without limitation, eMolecules, Zinc,
MMsINCdatabase,
Pubchem, Chemspider, chEMBL, Chemical Structure Lookup Service, CoCoCo, Broad
Institute compound collection(s), NIH Molecular Libraries Probe Production
Centers
Network (MLPCN), Joint European Compound library at the European Lead factory,
ScreeningPort at Fraunhofer Institute for Molecular Biology and Applied
Ecology (IME),
Microsource Spectrum collection (contains human approved/trialled drugs),
and/or by a
compound collection/library generated by "diversity-oriented synthesis" and/or
by one of the
art. A combinatorial library, generated by combinatorial chemistry, may be
used, wherein
these terms are well known in the art (e.g. EP0774464, U55798035, U55789172,
US5751629; and refer to patents with the combinatorial chemistry: sub-class
"C40B" in the
International Patent Classification; refer GLARE software, available on
sourceforge.net
website, for combinatorial library design).
A method to find antibody embodiment(s) of this invention is to raise
antibodies against an
ATP synthase component(s), and/or the entirety of ATP synthase, and then assay
each in one
or more of the aforementioned assays, looking for the ability to
preferentially/specifically
inhibit FIE) ATP hydrolysis as compared to FIFO ATP synthesis. An invention
embodiment is
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CA 3050553 2019-07-25
to administer a nucleotide sequence coding for such an antibody to a subject,
optionally by
gene therapy, optionally wherein this antibody coding gene is integrated into
the subject's
genome in one or more cells, optionally into the subject's mitochondrial DNA
(mtDNA) in
one or more cells. In an embodiment, one or more antibody embodiments of this
invention,
and/or one or more nucleotide sequences encoding one or more of such
antibodies, are
administered to a subject to convey to them therapy/enhancement, optionally
cancer
treatment/amelioration/prevention/combat, optionally wherein one or more of
said nucleotide
sequences are incorporated into the subject's genome, and/or mitochondrial
DNA, in one or
more of their cells, optionally wherein the expression of this nucleotide
sequence, to
protein(s), is limited to a certain cell type/tissue type/organ/area/sub-
section of the subject,
optionally by the character of the promotor region incorporated with the
protein(s) coding
sequence and/or by where the sequence is targeted to insert into the genome
and/or by where
in the subject the nucleotide sequence (optionally in a vector) is introduced
and/or by the
nature of the vector selected. Incidentally, an invention embodiment is for an
ATP synthase
.. component(s)/entirety to be administered to a subject, optionally via
intravenous
administration, wherein this acts as an epitope in the subject, wherein the
subject produces
antibodies against it, which then convey therapy/enhancement to the subject.
What the terms
"antibody" and "antibodies" can refer to, and how to produce them
(illustrative e.g. refer
US2008/0089950A1, Methods and compositions for modulating the immune system
and uses
thereof, Lan Bo Chen is one of the inventors, also refer to the patents and
publications that it
cites), is well known in the art and can include, without restriction,
monoclonal antibodies,
multispecific antibodies, human antibodies, humanized antibodies, camelised
antibodies,
chimeric antibodies, single domain antibodies, single-chain FVS (ScPv), single
chain
antibodies, Fab fragments, F(abl) fragments, disulfide-linked FVs (sdFv), and
anti-idiotypic
(anti-Id) antibodies, and epitope-binding fragments of any of the above. In
particular,
antibodies include immunoglobulin molecules and immunologically active
fragments of
immunoglobulin molecules, i.e., molecules that contain an antigen binding
site.
Immunoglobulin molecules can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and
IgY), class
(e.g., IgGI, IgG2, IgG3, IgGa, IgAi and IgA2) or subclass.
The present invention isn't limited to a particular type of compound. In
certain embodiments,
a compound of the present invention can be, but isn't limited to, an inorganic
molecule,
organic molecule, small organic molecule, small molecule, drug compound, large
molecule,
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CA 3050553 2019-07-25
nucleic acid, LNA (locked nucleic acid), polynucleotide, oligonucleotide, DNA
molecule,
gene, protein coding sequence of DNA and/or RNA, plasmid, virus, morpholino,
RNA
molecule, mRNA, hairpin RNA, siRNA (small interfering RNA), miRNA, antagomir,
ribozyme, aptamer, amino acid, amino acid chain, peptide, cyclic peptide,
bicyclic peptide,
tricyclic (or higher number of cycles) peptide, peptidomimetic, polypeptide,
protein, fusion
protein, glycopeptide, glycoprotein, antibody, antibody fragment, antibody-
drug conjugate,
PNA (peptide nucleic acid), lipid, sugar, carbohydrate.
CANCER TYPES PARTICULARLY TARGETED BY THIS INVENTION
Leukemia cancer cell line examples in Figure 8 show that Compound 6b can
inhibit cancer
proliferation strongly at 10 M and then nearly not at all at 100 M. As
disclosed earlier,
lower [6b] switches cancer cells into greater OXPHOS use/rate, exerting anti-
cancer activity,
but higher [6b] decreases OXPHOS rate, exerting less anti-cancer activity.
Extrapolating this
data, even greater [6b] may assist cancer and increase its proliferation rate.
And if a cancer
has appropriate adaptations and is using OXPHOS greatly in the first place,
without 6b
administration, then 6b administration may only ever help and never hinder
that cancer.
Compound 6b, and other FIFO ATP hydrolysis inhibiting compounds of this
invention, are
best administered to cancers showing signs of glycolytic rather than oxidative
metabolism.
For example, those that show up, at least before treatment by this invention,
in positron
emission tomography (PET) imaging using fluorine-18 (18F) fluorodeoxyglucose
(FDG), 18F-
FDG PET [71].
Particularly vulnerable to compounds of this invention: cancers that exhibit
the Warburg
effect (i.e. that produce ATP primarily by glycolysis, rather than oxidative
phosphorylation,
even in abundant 02), highly glycolytic cancers (which metabolize glucose
and/or glutamine
to lactate rather than metabolizing one or both fully with the use of
oxidative
phosphorylation) and cancers that reside in hypoxia, which forces them to
produce ATP
primarily by glycolysis. As explained in a preceding section, the imidazole
containing
molecules of this disclosure, with their uncoupling capability, can also
attack cancers that
reside in hypoxia, which use high IFI expression to enable oxidative
phosphorylation at low
[02]. Many cancers reside in hypoxia as tumours are often hypoxic.
So, if a cancer is highly glycolytic, either because of the Warburg effect
(inherent glycolytic
metabolism, regardless of [02]) or because of residing in hypoxia (imposed
glycolytic
CA 3050553 2019-07-25
metabolism, because of low [02]), or uses oxidative metabolism but resides in
hypoxia
(survival enabled by high IF, expression), it will be
treated/ameliorated/prevented/combated
by a compound of this invention. How to identify such cancers?
Cancers exhibiting the Warburg effect, or that have an imposed (by low [02])
glycolytic
metabolism, are those that show up in positron emission tomography (PET)
imaging using
18F-FDG PET, optionally integrated with computed tomography (CT) [71]. FDG is
a glucose
analogue and glycolytic cancers take up more FDG than their surrounding tissue
because
glycolysis is an inefficient metabolism of glucose (yielding only ¨2 ATP per
glucose
compared to ¨30 ATP per glucose yielded by aerobic respiration [1-21) and so
they must
uptake more glucose to obtain even the equivalent energy yield to nearby
normal cells, which
are using oxidative metabolism, as most normal cells do. So, if a cancer
presents in this FDG-
PET diagnostic (higher glucose uptake than surround), it is susceptible to a
compound of this
invention. Highly glycolytic cancers also release much lactate. So, if a
patient has a high
blood lactate level, noticeably above the normal non-pathological range, as
clear to someone
of the art, then their cancer is susceptible to a compound of this invention.
Higher lactate
levels in and around the cancer or tumour (than surrounding tissue) can also
be detected using
imaging technologies, for example Magnetic Resonance Spectroscopy ('H-MRS)
or
chemical exchange saturation transfer magnetic resonance imaging (CEST MRI)
[72], or
other imaging modalities and methods of the art. So, if a cancer presents
(higher [lactate] than
surround) in a lactate imaging diagnostic it is susceptible to a compound of
this invention.
Cancer release of lactic acid acidifies its extracellular space and this
acidification can be
detected by imaging modalities, well known to those of the art e.g. [73-74],
and if a cancer
can be discriminated from its surrounding tissue by this method then it is
susceptible to a
compound of this invention. An oxygen-sensitive chemical probe can be used to
obtain 3D
maps of tissue p02 [75], and if a cancer is shown to reside in notable hypoxia
then it is
susceptible to a compound of this disclosure, because it is either glycolytic
or likely using
high IF, expression to enable oxidative metabolism, both of which make it
susceptible to a
compound of this invention. Imaging technologies can be integrated to improve
signal to
noise e.g. [75] integrate p02 and lactate imaging. Such integration can give
added
information: for example, a cancer producing much lactate in a high p02
environment is
exhibiting the Warburg effect because it is heavily utilising glycolytic
metabolism in
abundant 02. Cancer gene expression markers and indicators of the Warburg
effect, well
known to those of the art e.g. [18-20], specify that a cancer is susceptible
to a compound of
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CA 3050553 2019-07-25
this invention, wherein the cancer's genetic material can be retrieved by
biopsy, surgery,
cancer cells or parts circulating in the bloodstream or some other method of
the art.
If a cancer uses oxidative phosphorylation (OXPHOS) rather than glycolytic
metabolism, and
it does not already improve its OXPHOS efficiency by high IF' gene expression
(which many
cancers do e.g. refer [23]) then a compound of this invention, by
preferentially inhibiting FiFo
ATP hydrolysis, will confer this efficiency gain and actually assist, rather
than harm, this
cancer. How to identify these cancers? A cancer's IF] gene expression, and
particularly its
gene expression ratio of IF] to a core ATP synthase sub-unit (e.g. ATP6), is
informative.
More so if compared to the corresponding gene expressions in a normal cell of
its host tissue,
so detecting difference from normal. If a cancer uses oxidative, rather than
glycolytic,
metabolism and does not have an appreciably higher IF' (or IFI/ATP6 ratio)
gene expression
than its corresponding normal tissue then it isn't prudent to use a compound
of this invention
for cancer therapy. More simply, it is best to use a compound(s) of this
invention against
highly glycolytic cancers and some (non-limiting) imaging methods have been
described
herein to identify these.
This invention discloses a method of using a compound(s) that preferentially
inhibits the
ATP-hydrolysing mode of ATP synthase, for example a compound(s) of Formula
(I), (II),
(III), (IV), (V) or (VII), or a pharmaceutically-acceptable salt, solvate,
hydrate or prodrug
thereof, to treat/ameliorate/prevent/combat a cancer that preferentially uses
glycolytic rather
than oxidative metabolism, for example a cancer exhibiting the Warburg effect,
and discloses
methods to identify these cancers. Identification methods specified are to
illustrate the
invention and not to limit its scope: this invention encompasses all methods
to identify
glycolytic cancers, in order to identify cancers most amenable to treatment by
a compound(s)
of this invention.
So, innovatively and usefully, compounds of this disclosure are selected for
anti-cancer
therapy by metabolic feature of the cancer, which belie how the cancer
survives and
proliferates, and its weaknesses, weaknesses that compounds of this disclosure
attack, rather
than the typical, often too arbitrary, often unhelpful, allocation by tissue
type, which is the
present standard in the art. A diversity of cancers, from different tissues,
will be susceptible
to compounds of this invention, especially the most dangerous: glycolytic
cancers, with high
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CA 3050553 2019-07-25
lactate efflux, often have the worst prognosis [18-20, 27]. Experimental data
of this
disclosure shows that compounds of this invention are effective against many
cancers.
Compounds of the present invention treat tumour growth, treat metastasis,
treat metastatic
cancer, treat non-metastatic cancer, treat tumour implantation, are useful as
an adjunct to
chemo-/radio- therapy, treat cancers including, but not limited to, advanced
malignancy,
multiple brain metastase, poor prognosis malignant brain tumor, metastatic
hepatocellular
carcinoma, mesothelioma, malignant melanoma, malignant mesothelioma, malignant
pleural
effusion mesothelioma syndrome, neuroendocrine tumor, amyloidosis, meningioma,
hemangiopericytoma, chondrosarcoma, Ewing's sarcoma, malignant fibrous
histiocytoma of
bone/osteosarcoma, osteosarcoma, rhabdomyosarcoma, heart cancer, brain cancer,
astrocytoma, neuronal & mixed neuronal-glial tumors, glioma, brainstem glioma,
pilocytic
astrocytoma, ependymoma, HPV induced tumors, primitive neuroectodermal tumor,
craniopharyngioma, cerebellar astrocytoma, cerebral astrocytoma, malignant
glioma,
recurrent malignant glioma, medulloblastoma, neuroblastoma, schwannoma,
oligodendroglioma, anaplastic oligodendroglioma, pineal astrocytoma,
anaplastic
astrocytoma, pituitary adenoma, visual pathway and hypothalamic glioma,
glioblastoma,
glioblastoma multiforms, breast cancer, invasive ductal carcinoma, ductal
carcinoma in situ
(DCIS), invasive lobular carcinoma, tubular carcinoma, invasive cribriform
carcinoma,
medulloblastoma, medullary carcinoma, male breast cancer, phyllodes tumor,
inflammatory
breast cancer, adrenocortical carcinoma, islet cell carcinoma, multiple
endocrine neoplasia
syndrome, parathyroid cancer, pheochromocytoma, thyroid cancer, medullary
thyroid
carcinoma, papillary thyroid carcinoma, follicular thyroid carcinoma, merkel
cell carcinoma,
intraocular melanoma, retinoblastoma, ocular neoplasm, anal cancer, appendix
cancer,
cholangiocarcinoma, carcinoid tumor, colon cancer, extrahepatic bile duct
cancer, gallbladder
cancer, gastric (stomach) cancer, gastrointestinal cancer, gastrointestinal
carcinoid tumor,
gastrointestinal stromal tumor (GIST), hepatocellular cancer, pancreatic
cancer, rectal cancer,
bladder cancer, cervical cancer, endometrial cancer, extragonadal germ cell
tumor, ovarian
cancer, ovarian epithelial cancer (surface epithelial-stromal tumor), ovarian
germ cell tumor,
penile cancer, renal cell carcinoma, renal pelvis and ureter, transitional
cell cancer, prostate
cancer, androgen independent prostate cancer, androgen dependent stage IV non-
metastatic
prostate cancer, hormone-refractory cancer, hormone-insensitive prostate
cancer,
chemotherapy-insensitive prostate cancer, testicular cancer, gestational
trophoblastic tumor,
ureter and renal pelvis, genitourinary cancer, transitional cell cancer,
urethral cancer, uterine
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CA 3050553 2019-07-25
sarcoma, vaginal cancer, vulvar cancer, wilms tumor, esophageal cancer, head
and neck
cancer, nasopharyngeal carcinoma, oral cancer, oropharyngeal cancer, paranasal
sinus and
nasal cavity cancer, pharyngeal cancer, salivary gland cancer, hypopharyngeal
cancer, acute
biphenotypic leukemia, acute eosinophilic leukemia, acute lymphoblastic
leukemia (ALL),
acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute myeloid
dendritic
cell leukemia, karotype acute myeloblastic leukemia, primary myelofibrosis,
myelodysplastic
syndromes (MDS), myeloid sarcoma, myeloproliferative neoplasms (MPNs),
lymphoma,
AIDS-related lymphoma, anaplastic large cell lymphoma, angioimmunoblastic T-
cell
lymphoma, B-cell prolymphocytic leukemia, low grade follicular lymphoma,
Burkitt's
lymphoma, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia,
cutaneous
B-Cell lymphoma, cutaneous T-cell lymphoma, diffuse large B-cell lymphoma,
follicular
lymphoma, hairy cell leukemia, hepatosplenic T-cell lymphoma, Hodgkin's
lymphoma, non-
Hodgkin's lymphoma, hairy cell leukemia, intravascular large B-cell lymphoma,
large
granular lymphocytic leukemia, lymphoplasmacytic lymphoma, lymphomatoid
granulomatosis, mantle cell lymphoma, marginal zone B-cell lymphoma, mast cell
leukemia,
mediastinal large B cell lymphoma, multiple myeloma/plasma cell neoplasm,
myelodysplastic syndromes, mucosa-associated lymphoid tissue lymphoma, mycosis
fungoides, nodal marginal zone B cell lymphoma, non-Hodgkin lymphoma,
precursor B
lymphoblastic leukemia, primary central nervous system lymphoma, primary
cutaneous
follicular lymphoma, primary cutaneous immunocytoma, primary effusion
lymphoma,
plasmablastic lymphoma, Sezary syndrome, splenic marginal zone lymphoma, T-
cell
prolymphocytic leukemia, basal-cell carcinoma, melanoma, skin cancer (non-
melanoma),
bronchial adenomas/carcinoids, small cell lung cancer, mesothelioma, Non-Small
Cell Lung
Cancer (NSCLC), pleuropulmonary blastoma, adenocarcinoma, rectal
adenocarcinoma,
unresectable colorectal carcinoma, laryngeal cancer, thymoma and thymic
carcinoma,
peritoneal carcinoma, peritoneal cancer, papillary serous carcinoma, AIDS-
related cancers,
Kaposi sarcoma, epithelioid hemangioendothelioma (EHE), desmoplastic small
round cell
tumor, leiomyoma, leiomyosarcoma, Liposarcoma, fallopian tube cancer,
smoldering
myeloma, indolent myeloma, Waldenstrom's macroglobulinemia, fibrodysplasia
ossificans
progressive, breast carcinoma, non-small cell lung carcinoma, ovarian
carcinoma, pancreatic
carcinoma, prostate carcinoma, colorectal carcinoma, squamous cell carcinoma,
hepatocellular carcinoma benign prostatic hyperplasia (BPH) and polycystic
ovary syndrome,
a neoplasm disclosed by the International Classification of Diseases (ICD) in
ICD-10 Chapter
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CA 3050553 2019-07-25
II: Neoplasms (World Health Organisation, WHO) and/or the International
Classification of
Diseases for Oncology (WHO).
The compounds of the present invention treat cancers including, but not
limited to, those that
originate in the testis, cerebral cortex, skin, fallopian tube, parathyroid
gland, small intestine,
large intestine, kidney, skeletal muscle, duodenun, spleen, epididymis, bone,
bone marrow,
lymph node, adrenal gland, esophagus, thyroid gland, heart muscle, tonsil,
lung, prostate,
rectum, anus, adipose tissue, colon, stomach, cervix, gallbladder, seminal
vesicle, breast,
ovary, endometrium, smooth muscle, salivary gland, pancreas, urinary bladder,
blood, brain,
.. gum, mouth, throat, head, liver, nasopharynx, neck, tongue, uterus, penis,
vagina, chest, eye,
head, neck.
Local administration of a compound(s) of this invention, optionally for cancer
treatment
In some invention embodiments a compound(s) of this invention, optionally a
compound(s)
of Formula (I), (II), (III), (IV), (V), (VI), (VII), or a pharmaceutically-
acceptable salt, solvate,
hydrate or prodrug thereof, is administered to a subject locally rather than
systemically,
optionally to convey therapy, optionally to treat/ameliorate/prevent/combat
cancer in a
subject, optionally wherein the local administration is to the cancer(s)
itself. For non-limiting
.. example, wherein the local administration is to a skin cancer(s) and/or pre-
cancer, optionally
basal-cell skin cancer (BCC), squamous-cell skin cancer (SCC), melanoma,
dermatofibrosarcoma protuberans, Merkel cell carcinoma, Kaposi's sarcoma,
keratoacanthoma, spindle cell tumor, sebaceous carcinoma, microcystic adnexal
carcinoma,
Paget's disease of the breast, atypical fibroxanthoma, leiomyosarcoma,
angiosarcoma,
hemangioma, Melanocytic nevus, Bowen's disease, Actinic keratoses, optionally
administered via a
liquid/solution/cream/lotion/ointment/emulsion/foam/spray/patch/transdermal
patch/adhesive
bandage/time release technology or some other drug administration route known
to one of the
art. Skin cancer is the most prevalent cancer globally. This local drug
administration can
locally reduce Fi Fo ATP hydrolysis, thence FIFO ATP synthesis, oxidative
phosphorylation
rate and metabolic heat generation, which is not detrimental when ambient
temperature is
37 C, and not detrimental when ambient temperature is lower because heat
transfer from the
rest of the body, especially via blood flow, maintains the drug administered
area at or near
37 C.
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CA 3050553 2019-07-25
CANCER IMAGING
An FiFo ATP hydrolysis inhibitor compound(s) of this invention, for example a
compound(s)
of Formula (I), (II), (III), (IV), (V) or (VII), has greater binding affinity
for ATP synthase
operating in reverse than forwards. Thence such a compound(s)
disproportionally
accumulates in cancer cells because, as disclosed herein, cancers
disproportionally utilise
ATP synthase in reverse, as compared to normal cells. Thus, when the
compound(s) is
labelled, for example by "C or some other radionuclide incorporation, it can
be used for
cancer imaging, for example by positron emission tomography (PET). Example
embodiments
are one or more of 11C, 18F, 13N, 150, 124I incorporated into a compound of
this invention at a
greater than natural abundance, most preferably incorporated at their
corresponding
position(s) in the compound(s) e.g. "C in place of 12C, 13N in place of 14N,
1241 at halogen
designated position(s) etc. All radionuclide substitutions of corresponding
atoms in Formula
(I-V, VII) are contemplated and componentry to the present invention, as is
their use for anti-
cancer imaging and/or therapy. Alternative embodiments include iodine
incorporated at a
position(s) designated halogen in one or more compounds of Formula (I-V, VII),
and this
compound(s) is then used for iodinated x-ray contrast imaging of cancer.
Alternatively 1231 is
incorporated at halogen position(s) of one or more compounds of Formula (I-V,
VII), and this
compound(s) is used to image cancer using single photon emission computed
tomography
(SPECT). If a cancer shows up (above background tissue) in one or more of
these imaging
modalities, it is a good signal that this cancer will be responsive to a
compound of this
invention for anti-cancer therapy.
Given this asymmetric accumulation of compounds of Formula (I-V, VII) into
cancer cells,
radionuclides incorporated into one or more of these compound(s) can
disproportionally
damage cancer, and thence be used for radiotherapy. To illustrate, tritium at
greater than
natural abundance at one or more places on a compound(s) of this invention,
wherein this
compound(s) is used for anti-cancer therapy. Alternatively, or in addition,
1251 and/or 1311
, incorporated, optionally at a position(s) designated halogen in Formula
(1-V, VII).
Optionally, radionuclide imaging is performed prior to radionuclide therapy
(radiotherapy)
with compounds of this invention.
A boronated compound(s) of this invention, for example a compound(s) of
Formula (I), (II),
(III), (IV), (V) or (VII), substituted at one or more positions with 1 B, for
use in neutron
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CA 3050553 2019-07-25
capture therapy, optionally for an anti-cancer use, is componentry to the
present invention, as
is a method wherein 157Gd is used in place of ' B.
Compound(s) of this invention treat/ameliorate/prevent/combat non-cancerous
.. proliferative disorders
An embodiment of this invention is a method in which a subject takes or is
administered an
effective amount of a compound(s) of this invention, for example a compound(s)
of Formula
(I), (II), (III), (IV), (V) or (VII) or another compound(s) that selectively
inhibits FiFo ATP
hydrolysis, or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug
thereof, to
treat/ameliorate/prevent/combat a non-cancerous proliferative disorder(s)
including, to
illustrate, without restriction, hyperproliferative autoimmune disorders,
hyperplasia,
epidermal hyperplasia, dysplasia (e.g. epithelial dysplasia), nodule(s),
wart(s), papilloma(s),
squamous cell papilloma, genital wart(s), condyloma(s), condyloma acuminatum,
cyst(s),
polyp(s) {including, without restriction, digestive, colorectal, endometrial,
cervical, nasal,
.. laryngeal, inflammatory fibroid polyp[s])}, inherited/hereditary
(including, without
restriction, Familial adenomatous polyposis, Peutz¨Jeghers syndrome, Turcot
syndrome,
Juvenile polyposis syndrome, Cowden disease, Bannayan¨Riley¨Ruvalcaba syndrome
{Bannayan-Zonana syndrome}, Gardner's syndrome) and non-inherited (non-
restrictive e.g.
Cronkhite¨Canada syndrome) polyposis syndromes, benign tumour, adenoma, organ
enlargement by hyperplasia, Cushing's disease (enlarged adrenal cortex by
hyperplasia),
congenital adrenal hyperplasia, hyperplasia of breast, atypical ductal
hyperplasia, intraductal
papillomatosis, fibroadenomas, fibrocystic changes, hemihyperplasia, focal
epithelial
hyperplasia, sebaceous hyperplasia, sebaceous adenoma, intimal hyperplasia,
unwanted/undesirable smooth muscle cell proliferation, smooth muscle cell
hyperplasia,
.. intimal smooth muscle cell hyperplasia, neointimal hyperplasia,
proliferative vascular
disorders, stenosis, stenosis because of cellular proliferation, vaginal
stenosis, stenosis in a
blood vessel, stenosis in a blood vessel because of cellular proliferation,
vascular occlusion,
restenosis, restenosis in a blood vessel that has been implanted with a stent,
in-stent
restenosis, post-angioplasty restenosis, systemic sclerosis, cirrhosis of the
liver, adult
.. respiratory distress syndrome, idiopathic cardiomyopathy, lupus
erythematosus, retinopathy
(e.g. diabetic retinopathy and/or other retinopathy[y/ies]), cardiac
hyperplasia, fibrosis,
pulmonary fibrosis, idiopathic pulmonary fibrosis, fibromatosis,
neurofibromatosis, renal
interstitial fibrosis, Cowden syndrome, hamartomas, choristomas, hemangiomas,
lymphangiomas, rhabdomyomas, lymphangiomatosis, cystic hygroma, trichilemmoma,
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sarcoidosis, neurosarcoidosis, aggressive fibromatosis, desmoid tumours,
unwanted/undesirable skin cell proliferation, hyperproliferative skin
disorder, psoriasis
(including, without restriction, plaque, guttate, inverse, pustular, napkin,
seborrheic-like, nail,
scalp and erythrodermic psoriasis), psoriatic arthritis, dactylitis,
seborrhoeic dermatitis,
dandruff, eczema, atopic dermatitis, rosacea, reactive arthritis (Reiter's
syndrome), pityriasis
rubra pilaris, hyperproliferative variants of disorders of keratinization
(e.g., without
restriction, actinic keratosis, senile keratosis, keratosis pilaris,
seborrheic keratosis),
scleroderma, and the like. Other examples include reproductive system-
associated disorders
such as benign prostatic hyperplasia, prostate enlargement, endometrial
hyperplasia, atypical
endometrial hyperplasia, benign endometrial hyperplasia, adenomyosis, atypical
polypoid
adenomyoma, endometriosis, endometriosis of ovary (endometrioma), endometrial
polyp(s),
polycystic ovary syndrome, ovarian cyst(s), cervical polyp(s), uterine
fibroid(s), uterine
hyperplasia, and the like. Moreover, proliferative smooth muscle disorders,
such as intimal
smooth muscle cell hyperplasia, which can lead to blockage in, for non-
limiting example, the
urethra, the bile duct, the airway, the bronchial airways of the lung and/or a
blood vessel(s),
particularly following biologically or mechanically mediated tissue injury.
One common type
of non-cancerous proliferative disorder is restenosis, such as that associated
with balloon
angioplasty and/or insertion of a stent. In subjects with obstructive coronary
artery disease,
abatement of the chest pain associated with blocked blood vessels can
sometimes be achieved
by insertion of a stent-equipped angioplasty balloon. Inflating the balloon
opens the blood
vessel and installs the stent to keep the blood vessel open after removal of
the balloon. The
benefit is often temporary, however, because stented blood vessels can become
re-blocked
due to cell growth in response to tissue injury from the insertion. This
process is termed
restenosis. An embodiment of this invention is an angioplasty balloon and/or
stent (and/or
.. other medical device located inside the subject, optionally this device
includes a reservoir, a
coating composition, a controlled release polymer matrix, or the like which
comprises the
compound(s), which it can release in vivo, optionally over days or weeks or
months or years,
wherein optionally the device has surface contours for location of
compound(s)) coated with,
and/or containing ("drug-eluting balloon", "drug-eluting stent"), a
compound(s) of this
invention, for example a compound(s) of Formula (I), (II), (III), (IV), (V) or
(VII) or another
compound(s) that selectively inhibits FIFO ATP hydrolysis, or a
pharmaceutically-acceptable
salt, solvate, hydrate or prodrug thereof, and a method of implanting this
stent (and/or other
medical device) in a subject, optionally to maintain blood flow in a blood
vessel in the
subject, optionally in co-therapy with ionizing radiation, optionally wherein
the medical
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device, e.g. stent, is bioresorbable. In one embodiment, a compound(s) of the
invention is
delivered locally to a treatment site by contacting the treatment site with a
medical device
that includes the compound(s) in a reservoir, coating composition or
controlled release
polymer matrix. For example, the compound(s) may be included in a coating
composition or
.. reservoir in a stent. Stents in all locations in a subject are componentry
to this invention,
including glaucoma drainage stents (reduce intraocular pressure), duodenal
stents, colonic
stents, pancreatic stents, biliary stents, bile duct stents, esophageal
stents, gastrointestinal
stents, prostatic stents, ureteral stents, urethra stents, coronary stents,
peripheral stents,
arterial stents, venous stents, vascular stents, pulmonary stents and the
like. Componentry to
this invention is any medical/aesthetic device that is coated with and/or
contains a
compound(s) of this invention, and a method of inserting/implanting this in a
subject,
wherein such medical devices include, without restriction, stents, pacemakers
(e.g., including
a controller implanted under the skin and an electrode extending therefrom to
the heart,
optionally wherein any/all parts of this device can be coated with a
compound(s) of this
.. invention), implantable cardioverter defibrillator (ICD), tissue
augmentation implants (e.g.
breast implants), catheters, arterio-venous grafts, by-pass grafts, balloons
used in the
vasculature, sheaths for veins and arteries, GORE-TEX surgical prosthetics,
artificial valves,
artificial hearts, artificial joints, structural implants (pins, screws,
plates, and the like), tooth
implants, chochlear implants, osmotic pumps, and the like, any medical device
wherein
.. placement of the device at a treatment site in the subject can place that
site at risk of
pathological cell proliferation in response to tissue injury associated with
placement of the
device (e.g., formation of scars, lesions, adhesions, and the like). The
coating composition
can serve as a controlled release vehicle for the therapeutic compound(s) to
be delivered at
the site of a lesion, and can be selected such that the compound(s) can be
released at a desired
rate in vivo. The compound(s), optionally in combination with a polymer, can
be applied by
any conventional means such as dip coating, roll coating, spray coating, spin
coating, vapour
condensation, and the like. In a particular embodiment, coating polymers
include silicones
(poly siloxanes), polyurethanes, thermoplastic elastomers in general, ethylene
vinyl acetate
copolymers, polyolefin rubbers, EPDM rubbers, and combinations thereof. The
coating
composition can be polymeric and can further be hydrophilic, hydrophobic,
biodegradable, or
non-biodegradable. Example are homopolymers, co-polymers (including block
copolymers
and graft copolymers), dendritic polymers, crosslinked polymers and the like.
Suitable
polymers include synthetic and natural polymers (e.g. polysaccharides,
peptides) as well as
polymers prepared by condensation, addition and ring opening polymerizations.
Also
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included are rubbers, fibers and plastics. Polymers can be hydrophilic,
amphiphilic or
hydrophobic. In one aspect, the polymers of the present invention are non-
peptide polymers.
The material for the polymeric coating composition can be selected from the
group consisting
of polycarboxylic acids, cellulosic polymers, gelatin, polyvinylpyrrolidone,
maleic anhydride
polymers, polyamides, polyvinyl alcohols, polyethylene oxides,
glycosaminoglycans,
polysaccharides, polyesters, polyurethanes, silicones, polyorthoesters,
polyanhydrides,
polycarbonates, polypropylenes, polylactic acids, polyglycolic acids,
polycaprolactones,
polyhydroxybutyrate valerates, polyacrylamides, polyethers, and mixtures and
copolymers of
the foregoing. Coating compositions prepared from polymeric dispersions such
as
polyurethane dispersions (BAYHYDROL, etc.) and acrylic acid latex dispersions
can also be
employed. Biodegradable polymers that can employed in the coating composition
include
polymers such as poly(L-lactic acid), poly(DL-lactic acid), polycaprolactone,
poly(hydroxybutyrate), polyglycolide, poly(diaxanone), poly(hydroxyvalerate),
polyorthoester, copolymers such as poly (lactide co-glycolide),
polyhydroxy(butyrate-co-
valerate), polyglycollide-co-trimethylene carbonate; polyanhydrides;
polyphosphoester,
polyphosphoester-urethane; polyamino acids; polycyanoacrylates; biomolecules
such as
fibrin, fibrinogen, cellulose, starch, collagen and hyaluronic acid; and
mixtures of the
foregoing. Biostable materials that can be employed in the coating composition
include
polymers such as polyurethane, silicones, polyesters, polyolefins, polyamides,
polycaprolactam, polyimide, polyvinyl chloride, polyvinylmethyl ether,
polyvinyl alcohol,
acrylic polymers and copolymers, polyacrylonitrile, polystyrene copolymers of
vinyl
monomers with olefins (such as styrene acrylonitrile copolymers, ethylene
methyl
methacrylate copolymers, ethylene vinyl acetate), polyethers, rayons,
cellulosics (such as
cellulose acetate, cellulose nitrate, cellulose propionate, etc.), parylene
and derivatives
thereof, and mixtures and copolymers of the foregoing. Another polymer that
can be that can
be employed in the coating composition is poly(MPC:LAM.:HPMAy:TSMA,) where w,
x,
y, and z represent the molar ratios of monomers used in the feed for preparing
the polymer
and MPC represents the unit 2-methacryoyloxyethylphosphorylcholine, LMA
represents the
unit lauryl methacrylate, HPMA represents the unit 2-hydroxypropyl
methacrylate, and
TSMA represents the unit 3-trimethoxysilylpropylmethacrylate. The coated
medical device,
e.g., stent, can be used to maintain patency of a blood vessel, e.g. coronary
artery, previously
occluded by thrombus and/or atherosclerotic plaque. The delivery of a
compound(s)
described herein can reduce the rate of in-stent restenosis. Particular
polymers can be those
which are water insoluble and hydrophilic, i.e. can form hydrogels. A hydrogel
is a
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CA 3050553 2019-07-25
composition which can absorb large quantities of water. Polymers which can
form hydrogels
are generally more biocompatible than other polymers and can be used in
devices which are
inserted into, for example, vascular systems. Platelets and proteins typically
deposit upon
insertion of polymer into a treatment, e.g., vascular site and can initiate a
cascade of events
.. leading to restenosis or injury. This process can be slowed or eliminated
with polymers that
form hydrogels, resulting in reduced risk of protein deposition and platelet
activation.
Polymers which form hydrogels are typically crosslinked hydrophilic polymers.
Further
descriptions and examples of hydrogels are provided in Hydrogels and
Biodegradable
Polymers for Bioapplications, editors Attenbrite, Huang and Park, ACS
Symposium Series,
No. 627 (1996), U.S. Pat. Nos. 5,476,654, 5,498,613 and 5.487,898, the
teachings of which
are incorporated herein by reference. Examples of hydrogels include
polyethylene
hydroxides, polysaccharides and crosslinked polysaccharides. A "controlled
release" polymer
matrix can be a polymer combined with an active agent, such as a compound(s)
of this
invention, so that the active agent is released from the material in a
predesigned manner. For
example, the active agent may be released in a constant manner over a
predetermined period
of time, it may be released in a cyclic manner over a predetermined period of
time, or an
environmental condition or external event may trigger the release of the
active agent, and the
like. In one embodiment, the controlled release polymer matrix includes a
polymer that is
biologically degradable, chemically degradable, or both biologically and
chemically
.. degradable. In another embodiment, the controlled release polymer matrix
includes a non-
degradable polymer. Examples of suitable polymers for a controlled release
polymer matrix
include the polymers used for polymer coating compositions. In one embodiment,
a
controlled release polymer matrix is a coating. In another embodiment, the
controlled release
polymer matrix is solid component that forms part of the structure of the
medical device. For
example, a portion (e.g., about 1%, about 5%, about 10%, about 20% or about
50%) of the
fibers that make up a vascular graft can be made of a controlled release
polymer matrix.
Examples of synthetic biodegradable polymers include polyanhydrides,
polyhydroxyacids
such as polylactic acid, polyglycolic acids and copolymers thereof,
polyesters, polyamides,
polyorthoesters, and some polyphosphazenes. Examples of naturally occurring
biodegradable
polymers include proteins and polysaccharides such as collagen, hyaluronic
acid, albumin
and gelatin. A compound(s) of this invention can be encapsulated within,
throughout, and/or
on the surface of the implant/device. The compound(s) is released by
diffusion, degradation
of the polymer, or a combination thereof. There are two general classes of
biodegradable
polymers: those degrading by bulk erosion and those degrading by surface
erosion. U.S.
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Patents that describe the use of polyanhydrides for controlled delivery of
substances include
U.S. Pat. No. 4,857,311, U.S. Pat. No. 4,888,176, and U.S. Pat. No. 4,789,724
to Domb and
Langer. The entire teachings of these patents are incorporated herein by
reference. Non-
biodegradable polymers remain intact in vivo for extended periods of time
(e.g., at least about
one or more years). Drug loaded into the non-biodegradable polymer matrix is
released by
diffusion through the polymers micropore lattice in a sustained and
predictable fashion,
which can be tailored to provide a rapid or a slower release rate by altering
the percent drug
loading, porosity of the matrix, and implant structure. Ethylene-vinyl acetate
copolymer
(EVAc) is an example of a nonbiodegradable polymer that has been used as a
local delivery
system for proteins and other micromolecules, as reported by Langer and
Folkman, Nature
(London) 263:797-799 (1976). Other non-biodegradable polymers include
polyurethanes,
polyacrylonitriles, and some polyphosphazenes.
AGING
An invention embodiment is a method in which a subject takes or is
administered an effective
amount of a compound(s) of this invention, for example a compound of Formula
(1), (II),
(III), (IV), (V) or (VII) or another compound that selectively inhibits F1F0
ATP hydrolysis, or
a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, to
treat/ameliorate/prevent/combat an accelerated aging disease or progeroid
syndrome
including, but not limited to, Werner syndrome, Bloom syndrome, De Barsy
syndrome,
Rothmund¨Thomson syndrome, Cockayne syndrome, xeroderma pigmentosum,
trichothiodystrophy, combined xeroderma pigmentosum-Cockayne syndrome,
restrictive
dermopathy, Wiedemann¨Rautenstrauch syndrome, Hutchinson¨Gilford progeria
syndrome
(progeria), any progeroid syndrome, Ataxia telangiectasia-like disorder 2, XFE
progeroid
.. syndrome, Muscular dystrophy, Muscular Dystrophy (Becker's, Duchenne, Limb-
Girdle),
Yamamoto's Muscular Dystrophy, Mandibuloacral dysplasia, Dilated
cardiomyopathy,
GAPO syndrome, Cutis laxia, Ehlers-Danlos syndrom, Lenz-Majewski hyperostatic
dwarfism, SHORT syndrome, Progressive external opthalmoplegia, Nester-
Guillermo
progeria syndrome, MDPL syndrome, Dyskeratosis congenital, Down syndrome
and/or to
treat/ameliorate/prevent/combat an aging associated disease or
disease/disorder of aging
(incidence increases with increased age/senescence) and/or an unwanted aspect
of aging
and/or a disease/disorder associated with reactive oxygen species (ROS, e.g.
elevated [ROS])
including, but not limited to, age-associated decline, degenerative diseases,
neurodegenerative diseases, amyotrophic lateral sclerosis (ALS), Parkinson's
disease,
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CA 3050553 2019-07-25
Multiple System Atrophy (MSA), Progressive Supranuclear Palsy (PSP), essential
tremor,
resting tremor, Alzheimer's disease, Huntington's disease, spinocerebellar
ataxias,
Friedreich's ataxia, dementia, frontotemporal dementia, chronic traumatic
encephalopathy,
memory loss, aged cognition, age/aging related cognitive decline/impairment,
hereditary
.. spastic paraplegia, Batten disease, polyglutamine diseases,
atherosclerosis, cardiovascular
disease, myocardial infarction, cerebrovascular disease, stroke, heart
failure, heart failure
with preserved ejection fraction, idiopathic pulmonary fibrosis, fibrotic
disease, pulmonary
disease, coronary artery disease, hypercholesterolemia, obesity, liver
disease, fatty liver
disease, lysosomal storage disease, amyloidosis, systemic sclerosis, kidney
disease, hepatic
cirrhosis, immunosenscence, clonal hematopoiesis, chronic obstructive
pulmonary disease
(COPD), hypertension, hypercholesterolemia, age-related thymic atrophy,
arthritis,
osteoarthritis, arthritis (Osteo-and Rheumatoid), Juvenile Rheumatoid
Arthritis (JRA),
Degenerative Disc Disease, Tendinopathy, Androgenetic Alopecia, male-pattern
baldness,
Idiopathic Pulmonary Fibrosis, systemic sclerosis, Chronic Obstructive
Pulmonary Disease,
Psoriasis, age-related loss of cardiac/pulmonary/cognitive/vision function,
diabetes, type 2
diabetes, andropause, sarcopenia, muscle weakness, cachexia, age-related
cachexia and/or
sarcopenia, aging frailty, frailty syndrome, osteoporosis, age-related macular
degeneration
(AMD, early/intermediate/late), neovascular/wet AMD, dry AMD, Geographic
atrophy (GA),
wet and dry AMD in the same eye(s), Stargardt's macular degeneration, Best
vitelliform
.. macular dystrophy, diabetic retinopathy, proliferative diabetic
retinopathy, diabetic macular
edema, age/aging-related eye disease, ophthalmological disease/disorder,
ocular disease,
vision loss, progressive vision impairment, myopia (short-sightedness),
degenerative myopia,
hyperopia (far-sightedness), accommodative dysfunction, glaucoma, cataract
formation,
retinal degeneration, progressive retinal degeneration, retinitis pigmentosa,
leber hereditary
optic neuropathy, Fuchs spot, Best's disease, Sorsby's fundus dystrophy,
hearing loss,
age/aging-related hearing loss, presbycusis, tinnitus, naive T cell shortage,
movement
disability, nonalcoholic fatty liver disease (NAFLD), nonalcoholic
steatohepatitis (NASH),
immunosenescence, respiratory/urinary tract infection (RTI/UTI) especially in
older/aged/elderly subjects, cancer etc., any age-related/correlated
disease/disorder/condition
.. including age-related/correlated cardiovascular, neurodegenerative, eye
diseases,
inflammatory diseases and conditions. Exemplary cardiovascular/cerebrovascular
diseases
and conditions include arteriosclerosis, coronary heart disease, arrhythmia,
heart failure,
hypertension, hypercholesterolemia, age-related thymic atrophy, orthostatic
hypotension,
myocardial infarction, angina pectoris, atherosclerosis, dyslipidemia, stroke,
heart disease,
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congenital heart disease, renal artery disease or stenosis, peripheral
vascular disease, chronic
obstructive pulmonary disease (COPD), chronic renal disease, renal failure,
and heart disease.
Exemplary neurodegenerative diseases include Parkinson's disease and
Alzheimer's disease.
Exemplary eye diseases and conditions include macular degeneration, Stargardt
disease,
cataracts, diabetic retinopathy and glaucoma. Exemplary inflammatory diseases
and
conditions include arthritis, such as rheumatoid arthritis. Additional
exemplary age-
associated diseases and disorders include cancer, ulcers, osteopetrosis,
progeria, weakness,
hearing loss, celiac disease, liver spots and type 2 diabetes. It is to be
understood that "age-
related" refers to diseases/disorders/conditions frequently associated with
aging, however, a
given subject need not be of advance age, but rather the methods, compounds
and
compositions of this invention can be used regardless of the subject's age.
Not only does an FiFo ATP hydrolysis inhibitor compound of this invention
treat/ameliorate/combat cancer in a subject, it also prevents cancer in a
subject, which is
distinct from many other cancer treatments (e.g. radiotherapy) which are a
drive to further
cancer, and so compounds of this invention are especially preferred for cancer
treatment in
children, who have enough lifespan left for secondary cancers, as a result of
radiotherapy for
example [76], to be a very severe concern. Also it is noteworthy that
compounds of this
invention both treat cancer and slow aging, whereas many present cancer
treatments
accelerate aging [77], causing greater incidence of age related disease(s) and
ailments.
An anti-aging compound does one or more of slowing/reversing aging,
slowing/reversing a
sign(s) of aging, extending lifespan and/or healthspan,
delaying/preventing/treating diseases
that have an increased incidence with age, such as the neurodegenerative
diseases, treating
accelerated aging diseases. Any anti-aging compound that targets/inhibits FIE)
ATP
hydrolysis is componentry to this invention, preferably those that
preferentially inhibit FiFo
ATP hydrolysis as compared to FIE) ATP synthesis, and most preferably those
that don't
inhibit FiFo ATP synthesis at all. This disclosure discloses numerous such
working examples,
many of which are also new compositions of matter, and discloses rationale and
methods to
find further working examples (e.g. SMP studies, looking for compounds that
inhibit FiFo
ATP hydrolysis more than FiFo ATP synthesis), which are, in turn, componentry
to this
invention and encompassed by this disclosure, for example for an anti-aging
use, or for other
disclosed use(s).
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ANTI-AGING SKIN CREAM
FIFO ATP hydrolysis inhibitor compound(s) of this invention slow aging but can
reduce body
temperature. An invention embodiment is to target an FIFO ATP hydrolysis
inhibitor
compound(s) to a part/area of the subject/body where slower aging is desired,
optionally for
aesthetic/cosmetic or medical/therapeutic desire or need. This body part or
area will have
slower aging and lesser heat production, but heat transfer from surrounding
body areas
(especially via blood flow) will maintain the temperature of this body
part/area at an
acceptable value. So, the temperature issue is mitigated and slower aging
endures in that body
part/area. An invention embodiment is a method in which a subject takes or is
administered
an effective amount of a compound(s) of this invention, for example a compound
of Formula
(I), (II), (III), (IV), (V) or (VII) or another compound that selectively
inhibits FIF0 ATP
hydrolysis, or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug
thereof, to
treat/ameliorate/prevent/combat skin aging, optionally administered to the
skin, optionally by
skin and/or subcutaneous injection/implant, optionally as a skin cream,
optionally to the face.
In another embodiment, administered to the scalp and/or hair, optionally in a
hair treatment,
optionally in a shampoo, to treat/ameliorate/prevent/combat hair follicle and
hair
aging/loss/greying/baldness. All means of applying a compound(s) of this
invention to the
skin, and/or scalp and/or hair are contemplated by, and componentry to, this
invention.
Cosmetic/aesthetic embodiments of the invention
An invention embodiment is an FIFO ATP hydrolysis inhibitor compound(s),
optionally at
least one compound of Formula (I), (II), (III), (IV), (V), (VI) or (VII), or a
salt, solvate,
hydrate, precursor, liposome or nanoparticle thereof, and/or a
composition/formulation
comprising one or compounds of Formula (I), (II), (III), (IV), (V), (VI) or
(VII), as the
entirety or a component/ingredient of a cosmetic, for example (without
restriction) a product
for one or more of cleansing, beautifying, promoting attractiveness and/or
altering the
appearance of a subject (e.g., without restriction, making a more
youthful/younger
appearance, reducing the appearance of lines and wrinkles, preventing/reducing
the signs of
aging/premature aging, making skin/hair look visibly younger), optionally
wherein the
product is intended to be rubbed/poured/sprinkled/sprayed on/introduced into
or otherwise
applied to the human body, for example (without restriction) a skin/hair care
product,
shampoo, antidandruff shampoo, conditioner, a product delivered by micro-
needling/Dermaroller/plasma-needling/DermaPen, hair tonic, hair colour, hair
dye, soap, soap
substitute, shower gel, bath oil, bubble bath, toothpaste, mouthwash,
moisturizer, emollient,
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skin/face/body/hair/moisturizing/anti-aging/wrinkle/Crow's-
feet/masking/whitening/age
spot/liver spot cream/oily cream/aqueous
cream/lotion/powder/spray/aerosol/butter/gel/hydrogel/oil/salve/liquid/alcohol/
emulsion/anhd
rous-
cream/stick/wax/ointment/foam/paste/solution/drop/gum/jelly/serum/scrub/mask/ba
lm,
.. stretch mark/cellulite/thigh cream/treatment, varicose vein cream,
chapstick, lipstick, lip
gloss, lip liner, lip plumper, lip balm, lip stain, lip conditioner, lip
primer, lip booster, lip
butter, make-up, makeup, mascara, mascara primer, eye shadow, eye liner,
eyebrow
pencil/cream/wax/gel/powder, foundation, concealer, bronzer, fake tan, rouge,
blush, blusher,
highlighter, setting spray, cleanser, skin cleanser, foaming wash, toner,
facial mask (non-
.. limiting e.g. clay-based masks, e.g. using kaolin clay or fuller's earth,
peel masks, sheet
masks), exfoliant, perfume, cologne, aftershave, shaving foam, beard balm,
fragrance,
deoderant, antiperspirant, hairstyling product(s), hairspray, nail polish,
massage oil, barrier
cream, sunscreen/sunblock/sun cream (e.g. offering protection against UVA
and/or UVB
radiation), spot/acne cream, wherein optionally the product also contains an
anti-
aging/rejuvenating/moisturizing/revitalizing/exfoliating/anti-oxidant
compound(s), and/or a
vitamin(s), and/or one or more ingredients in a cosmetic for sale in the
USA/Canada/European Union/Japan/China/Korea/Australia/Brazil, and/or one or
more
ingredients listed in the International Nomenclature of Cosmetic Ingredients
(INCI, INCI
names are developed by the International Nomenclature Committee (INC) and
published in
the International Cosmetic Ingredient Dictionary and Handbook), and/or one or
more
ingredients listed in CosIng (the European Commission database for information
on cosmetic
substances and ingredients),and/or one or more anti-aging
compounds/ingredients (e.g.,
without limitation, botulinum toxin, epidermal growth factor, Vitamin A, one
or more
Retinoids {e.g., without limitation, retinol, retinal, tretinoin [all-trans-
retinoic acid],
isotretinoin [13-cis-retinoic acid], alitretinoin [9-cis-retinoic acid],
etretinate, acitretin,
adapalene, bexarotene, tazarotene, seletinoid GI, one or more retinoid
complexes/salts/esters/ethers {e.g., without limitation, Retinyl palmitate},
one or more Alpha
Hydroxy Acids [AHAs], one or more Beta Hydroxy Acids [BHAs], Vitamin C,
Vitamin E,
Coenzyme Q10, one or more antioxidants, one or more peptides {e.g., without
restriction,
acetyl hexapeptide-3, acetyl hexapeptide-8}, one or more copper peptides e.g.
Copper peptide
GHK-Cu, Matryxil, Nicotinamide adenine dinucleotide (NAD ), Nicotinamide
mononucleotide [NMN], Nicotinamide riboside (NR), Nicotinamide (Nam),
Nicotinic acid
(NA), Nicotinic acid adenine dinucleotide (NaAD), Nicotinic acid
mononucleotide (NaMN),
platelet-rich plasma, rapamycin) and/or one or more compounds/ingredients to
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treat/ameliorate/prevent/combat hair loss (e.g., without limitation, one or
more 5a-Reductase
inhibitors {5-ARls, also known as dihydrotestosterone (DHT) blockers},
Finasteride,
Dutasteride, Epristeride, Saw palmetto extract, Serenoa repens extract,
Alfatradiol {also
known as 17a-estradiol}, one or more antiandrogens (e.g., without limitation,
steroidal
antiandrogens and non-steroidal antiandrogens), Bicalutamide, Bimatoprost,
Cyproterone
acetate, Flutamide, Ketoconazole, Latanoprost, Minoxidil, MK-434, Nepidermin,
Nonsteroidal antiandrogen, RU-58841, Spironolactone, Steroidal antiandrogen,
Topilutamide, Kopexil, Latanoprost, bimatoprost, Pinacidil, Diazoxide, one or
more
corticosteroids, IGF-1 {optionally in liposomes}), and/or one or more natural
product
extracts, and/or one or more of Acetone, Acetyl hexapeptide-3, Allantoin,
Aloe, Alpha
hydroxy acid, Aluminium zirconium tetrachlorohydrex gly, Argan oil, Azulene,
Behentrimonium chloride, Bimatoprost, Bisabolol, Canthaxanthin, Carnauba wax,
Castor oil,
Ceteareth, Cetyl alcohol, Cocamide DEA, Cocamide MEA, Cocamidopropyl betaine,
Cocamidopropyl hydroxysultaine, Cocoa butter, Conditioner, Copernicia Cerifera
(Carnauba)
Wax, Copper peptide GHK-Cu, Decamethylcyclopentasiloxane, Dihydroxyacetone,
Dioxalin,
Dipropylene glycol, Disodium cocoamphodiacetate, DMDM hydantoin, Erythrulose,
Ethyl
macadamiate, Ethylhexyl palmitate, Film-forming agent, Glycerol, Glyceryl
behenate, Glycol
distearate, Guaiazulene, Guanine, Hydrogenated jojoba oil, Hydrolyzed jojoba
ester,
lodopropynyl butylcarbamate, Isoceteth-20, Isopropyl jojobate, Isopropyl
myristate,
Isopropyl palmitate, Jojoba alcohol, Jojoba ester, Jojoba oil, Jojoba Wax PEG-
80 Esters,
Jojoba Wax PEG-120 Esters, Lapyrium, Macadamia oil, Malic acid, Marula oil,
Microbead,
Microcrystalline wax, Mineral cosmetics, Mineral oil, Myristamine oxide, Oleyl
alcohol,
denatured alcohol, Palmitoyl pentapeptide-4, Panthenol, Paraben, PEG-10
sunflower
glycerides, PEG-16 macadamia glycerides, PEG-80 Jojoba, PEG-120 Jojoba, PEG-
150
hydrogenated jojoba, Petroleum jelly, Polyacrylic acid, Polydimethylsiloxane,
Polyethylene
glycol propylene glycol cocoates, Polyquaternium, Polyquaternium-7, Propylene
glycol,
Quaternium-15, Rice bran wax, Sculptra, Selenium disulfide, Silicone,
Simmondsia
Chinensis (Jojoba) Seed Oil, Simmondsia chinensis (jojoba) seed powder, Sodium
laureth
sulfate, Sodium lauroamphoacetate, Sodium lauroyl sarcosinate, Sodium myreth
sulfate,
Spermaceti, Stearalkonium chloride, Stearamidopropyl dimethylamine, Sunflower
oil, Talc,
1-Tetradecanol, Tetramethyl acetyloctahydronaphthalenes, Tocopherol, 1-
Tridecanol,
Triethanolamine, Vitellaria, Zinc pyrithione, Zinc ricinoleate, Amiloxate, 4-
Aminobenzoic
acid, Avobenzone, Bemotrizinol, Benzophenone-n, Bisdisulizole disodium,
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Bisoctrizole, Cerium(IV) oxide, Cinoxate, Dibenzylideneacetone, Diethylamino
hydroxybenzoyl hexyl benzoate, Dioxybenzone, Drometrizole trisiloxane,
Ecamsule,
Ensulizole, Enzacamene, Ethylhexyl triazone, Homosalate, Iscotrizinol, Menthyl
anthranilate,
Mexenone, Octocrylene, Octyl methoxycinnamate, Octyl salicylate, Oxybenzone,
Padimate
A, Padimate 0, Polysilicone-15, Sulisobenzone, Titanium dioxide, Titanium
dioxide
nanoparticle, Trolamine salicylate, Umbelliferone, Zinc oxide, alkyl benzoate
C 12-C15,
allantoin, water, ascorbyl palmitate, butane, Butyrospermum parkii, shea
butter, cocoamide
dea, dodecanol, egg oil, hydroxyethyl cellulose, hydroxypropyl cellulose,
isobutane,
isopentane, lauryl glucoside, Polysorbate 20, propane, sodium hydroxide,
triethanolamine,
honey, shea butter, almond oil, argan oil, rosehip oil, beeswax, stevia,
glycerin, essential
oil(s), benzyl alcohol, dehydroacetic acid, Glyceryl Caprilate, Potassium
sorbate,
Caprylhydroxamic Acid, Caprylyl Glycol, Glycerin, Xanthan Gum, EDTA,
Emulsifying wax,
Olive oil, Evening primrose oil, Tocopheyl Acetate, Cetearyl Alcohol, Caprylyl
Glycol,
Phenoxyethanol, Hexylene Glycol, Glycyrrhiza Glabra (Licorice) Root Extract,
Sodium Hyaluronate.
Componentry to this invention is a compound of Formula (VII), e.g. IF1 (e.g.
from a human
and/or another mammalian species), in liposomes in a fluid gel formulation
(e.g., without limitation, as used in [78] for (IGF)-1) in use as a cosmetic,
optionally wherein
a more youthful/younger appearance is desired in a subject.
EYE AGING
Fi Fo ATP hydrolysis inhibitor compound(s) of this invention slow aging but
can reduce body
temperature. An invention embodiment is to target an FIR, ATP hydrolysis
inhibitor
compound(s) to one or both eyes of a subject, optionally by intravitreal
injection(s) and/or
eye drop and/or contact lens coating/solution (optionally wherein the contact
lens has little to
no refractive ability or wherein the contact lens is prescriptive to the
refractive defect/error of
the subject's eye(s)) and/or some other drug administration route/device to
the eye(s), known
or findable to those of the art, wherein the eye(s) then has slower aging and
lesser heat
production, but wherein heat transfer from surrounding body areas (especially
via blood
flow) maintains eye(s) temperature at acceptable value. So, the temperature
issue is mitigated
and slower aging in the eye(s) endures. An invention embodiment is a method in
which a
subject takes or is administered an effective amount of a compound(s) of this
invention, for
example a compound of Formula (I), (II), (III), (IV), (V) or (VII) or another
compound that
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selectively inhibits FiF0 ATP hydrolysis, or a pharmaceutically-acceptable
salt, solvate,
hydrate or prodrug thereof, optionally by a local drug administration route to
the eye(s) (e.g.
by an eye delivery route known or findable to those of the art e.g. an eye(s)
administration
route used for an FDA/EMA licensed/sanctioned drug(s) e.g. as described in the
patent/scientific literature e.g. refer [79, 80, 811 and the papers they cite
and the papers that
cite them), to treat/ameliorate/prevent/combat eye(s) aging and/or an eye
aging related
disease/disorder, including any eye disease/disorder whose likelihood of onset
increases with
age and/or worsens with age, including, without limitation, age-related
macular degeneration
(AMD, early/intermediate/late), neovascular/wet AMD, dry AMD, Geographic
atrophy (GA),
wet and dry AMD in the same eye(s), Stargardt's macular degeneration, Best
vitelliform
macular dystrophy, diabetic retinopathy, proliferative diabetic retinopathy,
diabetic macular
edema, vision loss, progressive vision impairment, myopia (short-sightedness),
degenerative
myopia, hyperopia (far-sightedness), accommodative dysfunction, glaucoma,
cataract
formation, retinal degeneration, progressive retinal degeneration, retinitis
pigmentosa, leber
hereditary optic neuropathy, Fuchs spot, Best's disease, Sorsby's fundus
dystrophy. In an
embodiment, one eye of the subject is treated and the other not (optionally
administered drug
vehicle control), optionally for a course of administrations over a period of
time, and the
anatomical/physiological/functional difference(s) between them is then
compared after some
period. Non-limiting example eye function tests are using the Snellen chart,
or LogMAR
chart, for visual acuity testing and/or the Amsler grid to investigate central
vision. In an
embodiment, a subject genetically predispositioned to age-associated eye
disease(s)/disorder(s), e.g. macular degeneration, optionally discovered by
genetic testing
and/or family history analysis, is administered a compound(s) of this
invention
prophylactically. When a compound(s) of this invention is administered by
intravitreal
injection(s), optionally antibiotic(s) eye drop(s) (alternatively/in addition
oral antibiotic(s)) is
administered one or more times on the same day and/or in the same week and/or
in the same
month. Given increasingly aging societies in many countries, more and more
people are
succumbing to macular degneration. It is noteworthy that there is presently no
treatment on
the market for dry AMD, which comprises 90% of macular degeneration cases,
affecting
millions globally. The projected number of people with age-related macular
degeneration in
2020 is 196 million, increasing to 288 million in 2040 [82].
EAR AGING
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FiFo ATP hydrolysis inhibitor compound(s) of this invention slow aging but can
reduce body
temperature. An invention embodiment is to target an FIFO ATP hydrolysis
inhibitor
compound(s) to one or both ears of a subject, optionally by intratympanic
and/or
intracochlear administration and/or trans-oval window delivery and/or by ear
drops and/or
some other drug administration route/device to the ear(s), known or findable
to those of the
art, wherein the ear(s) then has slower aging and lesser heat production, but
wherein heat
transfer from surrounding body areas (especially via blood flow) maintains
ear(s) temperature
at acceptable value. So, the temperature issue is mitigated and slower aging
in the ear(s)
endures. An invention embodiment is a method in which a subject takes or is
administered an
effective amount of a compound(s) of this invention, for example a compound of
Formula (I),
(II), (III), (IV), (V) or (VII) or another compound that selectively inhibits
FIE) ATP
hydrolysis, or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug
thereof,
optionally by a local drug administration route to the ear(s) (e.g. by an ear
delivery route
known or findable to those of the art e.g. an ear(s) administration route used
for an
FDA/EMA licensed/sanctioned drug(s) e.g. as described in the patent/scientific
literature e.g.
refer [83, 84] and the papers they cite and the papers that cite them), to
treat/ameliorate/prevent/combat ear(s) aging and/or an ear aging related
disease/disorder,
including any ear disease/disorder whose likelihood of onset increases with
age and/or
worsens with age, including, without limitation, age-related hearing loss,
presbycusis,
tinnitus.
BRAIN AGING
Neurodegenerative diseases have an aging component to their etiology [85] as
their onset is a
function of age (oxidative stress [85]). Indeed, all these diseases
(prototypical examples
include Parkinson's disease, dementia, Alzheimer's disease, amyotrophic
lateral sclerosis
{ALS}, Huntington's disease, Friedreich's ataxia, hereditary spastic
paraplegia) can be
thought of as the brain aging faster and dying before the rest of the body
(adult brain mass
decreases with age [86]). In our rapidly greying societies these diseases are
a demographic
time bomb. Indeed, beyond immeasurable personal suffering, they stand to
decimate whole
economies (healthcare spending becomes unsustainable percentage of GDP,
already ¨30% in
the USA). For example, nearly half of Americans, over 85, have dementia, which
in time is
an age that an increasing proportion of the population will surpass, it has no
cure and can be
completely debilitating, which strains families and communities [86]. Thus,
any treatment
that can slow brain aging, to make brain function last as long as the rest of
the body, will
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greatly assist in matching "healthspan" to lifespan, which is arguably the
Holy Grail in
modern medicine.
An invention embodiment is a method in which a subject takes or is
administered an effective
amount of a compound(s) of this invention, for example a compound of Formula
(I), (II),
(III), (IV), (V) or (VII) or another compound that selectively inhibits FIN
ATP hydrolysis, or
a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, to
treat/ameliorate/prevent/combat brain aging and neurodegenerative disease(s).
Optionally
wherein the compound(s) is disproportionally delivered to the brain or central
nervous system
(CNS), or to specific brain/CNS area(s) or cell type(s), by administration
route, strategy or
targeting. Illustratively, not restrictively, brain targeting had been shown
with exogenous
dopamine [87-88]. Preferred brain structures/cells/neurons to target are those
whose failure
drives a neurodegenerative disease e.g. dopamine neurons in the pars compacta
(in the
substantia nigra). There are few of them, only 7,200 in rat [89], and in
humans their number
decline by aging at 5-10% per decade [90], which is a predisposing drive to
Parkinson's
disease (PD). An invention embodiment is a method in which a subject takes or
is
administered an effective amount of a compound(s) of this invention, for
example a
compound of Formula (I), (II), (III), (IV), (V) or (VII) or another compound
that selectively
inhibits FIR) ATP hydrolysis, or a pharmaceutically-acceptable salt, solvate,
hydrate or
prodrug thereof, to treat/ameliorate/prevent/combat Parkinson's disease,
optionally wherein
the compound(s) is disproportionally administered to dopamine neurons in the
substantia
nigra. If a compound of this invention decreases their heat generation, heat
transfer from
neighbouring brain and/or body regions will substitute this heat.
LIFESPAN AND/OR HEALTHSPAN EXTENSION IN A SUBJECT BY
ADMINISTRATING A COMPOUND(S) OF THIS INVENTION
Maximal Tolerated Dose (MTD) study of a compound(s) of this invention
Three mice receive an initial dose of intravenous (i.v.) 10 mg/kg of drug. If
these mice
survive for 72 hours, the i.v. dose for the next cohort of three different
mice is increased,
whereas if one or more mice die, the i.v. dose for the next cohort of three
different mice is
decreased. And this schema is run iteratively. To illustrate, next dose level,
72 hours after
prior dose level, can be determined by the following scheme:
10 mg/kg, if no death, 30 mg/kg, if no death, 100 mg/kg
10 mg/kg if no death, 30 mg/kg, if death, 17 mg/kg
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mg/kg, if death, 3 mg/kg, if death, 1 mg/kg,
10 mg/kg, if death, 3 mg/kg, if no death, 5 mg/kg
But other schema can be developed by one of the art, involving different doses
and/or a
greater number of doses and/or using a different route of administration e.g.
oral (PO),
5 -- intraperitoneal (IP), intravenous (IV), subcutaneous (SC), intramuscular
(IM) or other. At
each dose level, animals are observed for the presence of acute toxic symptoms
(mortality,
convulsions, tremors, muscle relaxation, sedation, etc.) and autonomic effects
(diarrhea,
salivation, lacrimation, vasodilation, piloerection, etc.) during the first 60
minutes, again at 2,
24, 48 and 72 hours. Body weights are recorded pre-dose and at 72 hours after
dose.
An alternative MTD determination method that better conserves compound and
minimizes
the number of animals sacrificed: a single mouse is given a dose (IP, IV, SC,
IM or PO) of
400 mg/kg, a second mouse receives a dose of 200 mg/kg and a third mouse
receives a dose
of 100 mg/kg. The mice are observed for a period of 2 weeks. They are
sacrificed if they lose
-- more than 20% of their body weight or if there are other signs of
significant toxicity. If all 3
mice must be sacrificed or die, the next 3 dose levels (e.g. 50, 35 and 12.5
mg/kg) are tested
in a similar manner, whereas if only one or two dies, or needs to be
sacrificed, the next 3 dose
levels are between the highest shown safe dose thus far and the lowest
lethal/toxic dose
shown thus far. This process is repeated until a maximal tolerated dose (MTD)
is found.
A compound of this invention, a compound that preferentially inhibits FiFo ATP
hydrolysis
over FiFo ATP synthesis, for example a compound(s) of Formula (I), (II),
(III), (IV), (V) or
(VII), or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug
thereof has the
peculiarity that its MTD is higher if the animal is housed at 37 C instead of
normal room
temperature (-22 C). The MTD and/or LD50 and/or LD30 and/or LDio and/or No-
Observed-
Adverse-Effect Level (NOAEL) of a compound(s) of this invention, optionally
compound 7b,
is investigated and recorded at both temperatures, optionally at interim
temperature(s) also.
This information is then useful for implementing other example embodiment(s)
of this
invention. The MTD (or other drug dose safety measure) to use, the value to
use and apply in
-- designing a study, depends on what temperature(s) the animal(s) is to be
housed at in the
study. This different MTD at different temperature aspect to a compound(s) of
this invention
is more pronounced the smaller the animal e.g. more pronounced/important for
mice than
rats.
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Lifespan extension using a compound(s) of this invention
An example embodiment of this invention is to use a compound(s) of this
invention, a
compound that preferentially inhibits F1 F0 ATP hydrolysis over FIFO ATP
synthesis, for
example a compound(s) of Formula (I), (II), (III), (IV), (V) or (VII) or a
pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof in an animal (e.g. mouse)
lifespan study.
Illustratively, compound 7b is used in a mouse lifespan study. For non-
restrictive example,
300 six-week old female Mus Muscu/us C57BL/6 strain mice are sourced from a
commercial
vendor (e.g. Charles River Laboratories Inc., MA, USA). Alternatively more
mice can be
used to be able to statistically detect smaller percentage increases in
lifespan. In an alternative
embodiment male mice are used also, wherein a study with both sexes valuably
permits
gender difference(s) to be identified. However, males bring the additional
complexity of
fighting, which can lead to mice deaths (need more males than females because
some males
will be invariably lost to fighting). In other example embodiments another
mouse strain(s) is
used and/or genetically heterogeneous mice, which avoids genotype-specific
effects on
disease susceptibility. The mice are housed at 37 C, which is safe for mice
(refer [205]), by
setting the room/ambient temperature accordingly e.g. by placing their cage(s)
(3-5 mice per
cage) in a Plant-Growth/Veterinary or Animal Intensive Care (ICU) incubator(s)
set at this
temperature, wherein such incubators can be sourced from one or more of
Precision
Refrigerated Plant-Growth Incubators, Thermo Fisher Scientific, Darwin
Chambers Inc.,
Powers Scientific Inc, Brinsea Products Ltd., Lyon Technologies Inc., or
similar company,
wherein some of these companies even make custom incubator designs. The
electrical cost of
this temperature maintenance is reduced by running the study in a hot
geographical
location/country, Singapore. Mice are kept on a 12 hour light/dark cycle, in
40-70%
humidity, with corn cob bedding and have ad libitum sterilized/irradiated chow
(illustratively
AIN-93G standard diet or Purina 5LG6 or Purina 5001) and water. Preferably the
mice are
housed in a pathogen-free barrier environment (SPF conditions). One of the art
knows how to
successfully look after laboratory mice and there are well known guidelines
and guides
publically available. Mice are randomly allocated into two groups: 100 mice
are in the drug
treatment group, 200 mice are in the non-drug control group (twice more mice
in control than
drug group). Optionally a positive control (100 mice, no drug administered,
calorie restricted
diet) group is added. In some example embodiments the test drug is
administered to the mice
through drinking water/solution (in which case fluid intake of drug treatment
and control
groups is recorded). Indeed, there are compounds of this invention that are
orally bioavailable
e.g. 6b is 47% orally bioavailable in rats when administered in
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polyethyleneglycol:water:ethanol (1:1:1) solution [5]. Administering 6b as a
salt, e.g. 6b HC1,
increases its solubility, which is advantageous for oral administration via
drinking solution.
Alternatively, the drug (base and/or salt) is mixed in with previously
irradiated (sterilized)
chow, wherein the drug is 0.0001%, or 0.001%, or 0.01%, or 0.05% (recommended
starting
.. percentage for experimentation to find optimal percentage), or 1%, or 2%,
or 3% or another
percentage of chow weight, which is done by BioSery (Flemington, NJ, USA) or
TestDiet
Inc. (TestDiet, Richmond, IN, USA) or Dyets Inc. (Bethlehem, PA, USA) or a
similar
company/service, the drug content of chow is checked using HPLC, wherein this
chow is
produced every 2 months during the length of the study, and is (alongside
untreated chow)
stored refrigerated, is never permitted to exceed 40 C and is kept away from
light whenever
possible to ensure drug stability (the light/dark cycle in the mouse facility
is not altered).
Preferably, water and chow are warmed to 37 C before being accessible to the
mice. To
calculate how many mg/kg of drug any given mg/kg of drug in chow will deliver,
a 30 g
mouse consumes ¨5 g food/day [911(1/6 of body weight, which is an approximate
relation
that can be applied to younger and lighter mice also), so illustratively, to
deliver 40 mg/kg of
drug to mice per day requires drug to constitute 240 mg/kg (0.024%) of chow.
Optionally,
further mice can be sourced for multiple drug treatment groups, all with the
same number
(100) of mice, which differ in the percentage weight of chow that is the test
drug. So, that
mice of the different drug treatment groups are administered a different drug
dosage.
Optionally, the test drug is microencapsulated e.g. by Southwest Research
Institute (San
Antonio, Texas) using a spinning disk atomization coating process with the
enteric coating
material Eudragit S100 (Rohm Pharma). This thermoplastic coating material
increases the
drug fraction that survives the chow preparation process. Because the coating
material is
water soluble only in non-acidic conditions, the encapsulated drug is released
in the small
intestine rather than in the stomach. It is prudent to verify that the
compound retains activity
after incorporation into mouse chow and that therapeutic blood levels of the
drug can be
achieved (blood drawn from tail vein). Method(s) to record the amount of a
drug in blood is
well known to those of the art e.g. using HPLC with ultraviolet detection [92]
and/or LC-MS
and/or LC-MS/MS. If eating the chow kills the mice, iteratively reduce the
drug content of
the chow until the mice can safely survive eating the chow. To observe if
there is sufficient
drug in chow to cause a physiological effect, house the mice at 22 C and
record the rectal
temperature of the mice every 15 minutes and observe if their body temperature
falls. If so,
there is a working drug concentration in the chow. Optionally, increase the
drug content of
chow until the body temperature drop is sufficient to kill the mice in this 22
C study. Then
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check that this drug content of chow doesn't kill the mice when they are
housed at 37 C, if
not, proceed with the study, if so, reduce the dose until finding the largest
safe drug dose in
chow at 37 C. Either use this largest safe dose or some fractional function
(e.g. half e.g. 10%
e.g. another percentage) of it. In Figure 23 of this disclosure 40 mg/kg i.v.
of 6a was safe and
40 mg/kg i.v. of 6b wasn't safe at 22 C ambient temperature, 6b is 47% orally
bioavailable in
rats [5], and so the recommended oral starting dose of 6b is 40*2 = 80 mg/kg ;-
--; 0.05% weight
of chow, wherein this recommendation is extended to compound 7b also. However,
in other
embodiments a different chow drug percentage is used and one of the art will
be able to
experiment with different chow drug percentages to explore the best
positioning/compromise
between drug safety and maximal drug effect, wherein preferably a Maximal
Tolerated Dose
(MTD) study, as described elsewhere herein, would have been performed prior to
give further
information to guide this assessment. Methods to derive a drug dose to be used
in a drug trial,
if the drug's MTD/LD5o (drug dose that kills 50%)/LDio (drug dose that kills
10%) is known,
are well known in the art. In an embodiment, the No-Observed-Adverse-Effect
Level
(NOAEL) is found and used, or some selected fraction (e.g. 50%, 10% or other)
of it,
wherein most optimally the NOAEL is found for the route (e.g. oral) and method
(e.g. chow)
of drug administration used in the lifespan study. MTD studies are typically
single dose
studies whereas this will be a long term study with the drug being
administered frequently
over a long period and this distinction needs to be considered. Pilot studies
with small
numbers of mice can be very instructive for setting the parameters of larger
studies with
many mice. In an alternative embodiment the drug is (e.g. daily) administered
intravenously
(e.g. at tail vein) through a catheter wherein control mice also have a
catheter fitted and are
administered vehicle at the same frequency as test mice are administered
[drug+vehicle]. In
other embodiments the drug is administered by some other route/method of
administration.
Food intake (important to record because calorie restriction extends life
[93], so any
difference in food intake between the drug treatment and control groups needs
to be known;
food intake of drug treated mice will be less than control mice because the
drug renders their
metabolism more efficient requiring less food and so they will choose to eat
less food) and
body weight are measured on a biweekly or bimonthly basis for the duration of
the study.
Healthspan assays (e.g. as set out in [94, 95, 96] and/or assaying homeostatic
capacity and/or
observing heart rate variability and/or rotarod assay(s) and/or grip assay
and/or horizontal bar
assay and/or GSSG/GSH ratio and/or NAD/NADH ratio determination and/or one or
more of
the healthspan/fiinctional assays listed in another example embodiment of this
invention
and/or another assay of the art e.g. recording one or more of body
coordination, memory,
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learning, movement, cognitive function) can be performed at regular intervals,
especially as
the mice get older. During the study, the day that each mouse dies is recorded
and the study
ends when all mice have died. Survival curves are plotted using the
Kaplan¨Meier method,
which includes all available animals at each time point. Statistical analyses
is performed
using JMP IN (SAS, Cary, NC). The criteria for euthanasia is based on an
independent
assessment by a veterinarian, according to AAALAC guidelines and only cases,
where the
condition of the animal is considered incompatible with continued survival,
are represented in
the curves. Every animal found dead or euthanized is necropsied for pathology
score. At
study end, the mean, median and maximal lifespan is calculated separately for
drug treated
and control groups. Comparing the proportion of mice still alive in each group
at each age
when the pooled population reaches the 90% mortality point is also a useful
measure. The
data will show that 7b extends the lifespan of mice, especially if they are
maintained at 37 C.
This temperature dependence aspect can be shown by running the experiment
again, or in
parallel, wherein all the mice (drug treated and control) are kept at 22 C
rather than 37 C,
wherein there is a lower drug dose(s) with the drug treated group(s) at this
lower ambient
temperature (because the tolerated drug dose is lower at lower ambient
temperature), and
wherein, with lower drug dose, the lifespan extension isn't as great. In an
alternative
embodiment, when mice are first sourced they are older e.g. older (e.g. old)
mice are sourced
from the National Institute on Aging Aged Rodent Colony or from the Jackson
Laboratory
(USA, has 19.5 months old mice available, roughly equivalent to a 50 year old
human). This
means that the experiment will take less time to run, because the mice will
die sooner after
being received. But the increase in lifespan observed will be less. Another
way to shorten the
duration of the study is to use mice that undergo accelerated aging [97, 98,
99, 100] e.g.,
without limitation, Senescence Accelerated Mouse-Prone 8 (SAMP8) mice
(approximately
half the lifespan of normal laboratory mice; commercially available from
Harlan
Laboratories, Bicester, UK; also available from the Society for Senescence-
Accelerated
Mouse (SAM) Research, Japan [http://www.samrc.jp], as are further senescence
accelerated
mouse strains) and/or BubR1 progeroid mice [101] and/or XPD (e.g. XPDTTD
[102])
mutant mice (optionally carrying an additional mutation(s) in XPA and/or XPC)
[103, 104]
(mice with a Trichothiodystrophy [TTD] mutation in XPD, with XPC knocked out,
have
accelerated aging and only live 4-8 weeks) and/or XPC mutant mice [105, 106]
(commercially available from The Jackson Laboratory, Stock No: 010563) and/or
ERCC1
mutant mice (e.g. ERCC1-/- [107] e.g. ERCC1 Ai- mice carry a null mutation in
one allele and
a 7-amino acid truncation in the second allele, maximum lifespan is ¨6 months)
[108, 109,
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97] and/or Ku70 and/or Ku80 and/or Ku86 [110] and/or DNA-PKcs mutant mice
[111]
and/or Caspase-2 mutant mice (commercially available from The Jackson
Laboratory, Stock
No: 007899) [112, 113] and/or ICE mice (Induced Changes in Epigenome) and/or
some other
accelerated aging mouse model of the art. Some of these accelerated aging
mouse models, as
are others not mentioned but that can be found by one of the art, are
recognised models of
human accelerated aging diseases. Alternatively, to shorten the study
duration, one can use a
smaller and shorter lived mammal species than mice (20 g) e.g. the Common
Shrew (9 g) or
the even smaller Etruscan shrew (1.8 g). An embodiment of this invention is to
enter a
compound(s) of this invention into the Major Mouse Testing Program (MMTP)
and/or the
National Institute on Aging's Interventions Testing Program (ITP) and/or use
the
same/similar/inspired testing protocol for a lifespan study using a
compound(s) of this
invention, or another lifespan study protocol in the literature or a lifespan
study protocol
conceived by someone of the art, optionally after their reading lifespan
studies in the
literature e.g., without limitation, [92, 114, 115, 116, 117, 118, 119]. An
invention
embodiment is to enter a compound(s) of this invention, or result(s) from
using a
compound(s) of this invention, into a mouse/rodent or other animal lifespan
competition such
as the Methuselah Mouse Prize (MPrize) and/or Palo Alto Longevity Prize and/or
other/similar. A compound(s) of this invention extends lifespan by a direct
anti-aging effect
and also by an anti-cancer effect, reducing the incidence of, and by
treating/ameliorating/preventing/combating cancer. Given the established link
between
age/aging and neurodegenerative disease, a compound of this invention, which
slows aging as
shown by this example, has utility as a therapeutic for neurodegenerative
disease e.g.
(without restriction) for Alzheimer's disease and/or dementia. Illustratively,
Rapamycin
extends mouse lifespan [92, 117] and exerts therapy in a mouse model of
Alzheimer's disease
[120]. An accelerated mouse model of aging, SAMP8, is concurrently a mouse
model of
Alzheimer's disease [121].
Instead of, or in addition to, using mouse death as an endpoint in this study,
an
aging/mortality biomarker(s) can be used, e.g. one or more listed in the
database:
http://mortalitypredictors.org/ [139] e.g. walking speed. In this way,
compound(s) effect on
aging/mortality can be assayed before death. This is especially important for
human studies
with a compound(s) of this invention, or a pharmaceutically-acceptable salt,
solvate, hydrate
or prodrug thereof, optionally following on from mouse studies, given that
humans have a
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long lifespan and so a surrogate endpoint (change in a biomarker(s) e.g.
walking speed) to
assess the change in aging/mortality is preferable to awaiting lifespan data.
Healthspan assays show that a compound(s) of this invention slows aging,
including
brain aging, and treats/ameliorates/prevents/combats neurodegenerative
disease(s),
including Alzheimer's disease
APP/swePS1AE9 mice is a mouse model of Alzheimer's disease [140], available
from The
Jackson Laboratory (stock no: 004462). Senescence Accelerated Mouse-Prone 8
(SAMP8)
mice display a phenotype of accelerated aging, with associated cognitive
decline, and is a
.. mouse model of aging driving Alzheimer's disease and/or dementia [121],
available from
Harlan Laboratories (Bicester, UK). In an invention embodiment APP/swePS1AE9
mice (or
an alternative Alzheimer's disease mouse model {to illustrate and not
restrict: from the
Model-AD project and/or The Jackson Laboratory have a number of different
Alzheimer's
disease mouse models available [typically present learning deficit, from
variable age, many
including spatial learning deficit], or the PDAPP (also known as hAPP(J20)
transgenic mouse
model of Alzheimer's disease [120]}, or a mouse model of a different
neurodegenerative
disease e.g. a mouse model of Parkinson's disease, optionally sourced from The
Jackson
Laboratory) are used for the following study. In a different invention
embodiment SAMP8
mice (or an alternative accelerated aging mouse model) are used in the
following study. In a
.. different invention embodiment normal mice are used in the following study.
This study will
now be described with SAMP8 mice. At all places that SAMP8 is referred to, in
another
embodiment, "APP/swePS1AE9" is substituted in its place. At all places that
SAMP8 is
referred to, in a further embodiment, "normal" is substituted in its place.
Six-week old male
SAMP8 mice are sourced and randomly assigned to the following groups: 200
SAMP8 mice
are maintained on control chow (LabDiet 5015, TestDiet, Richmond, IN) and 100
SAMP8
mice are maintained on chow (LabDiet 5015) that contains a compound(s) of this
invention: a
compound that preferentially inhibits FIFO ATP hydrolysis over FIR) ATP
synthesis, for
example a compound(s) of Formula (I), (II), (III), (IV), (V) or (VII) or a
pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof. Illustratively, compound
7b of this
disclosure. How to prepare such chow has been disclosed for an earlier
invention example.
Food consumption and body weight are monitored during the study. In a further
example
embodiment there is more than one drug treated group, wherein these groups
differ in drug
dose given. Preferably all mice are housed at 37 C, as described for a prior
invention
example, and preferably the behavioural experiments are performed at 37 C
also, wherein
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preferably water in the water tests is also at 37 C. Mouse body weights are
measured
regularly. Behavioural tests are conducted every month. The study ends when
all mice die.
As the mice get older, drug treated SAMP8 mice start to outperform control
SAMP8 mice in
one or more of these tests, and/or a test variant(s), and/or a similar
test(s), and/or another
test(s) of mental/cognitive faculty(s) e.g. as found in the literature or as
modified by someone
of the art upon reading the literature: for the following test(s) the
experimenter is blinded to
which mice are drug treated and non-drug treated, preferably all tests are
videotaped for
parallel independent confirmatory analysis by another experimenter(s), wherein
automatic
computer software analysis is used where available/possible to aid analysis:
(1) Activity. The open field test is performed using MED Associates hardware
and Activity
Monitor software according to manufacturer's protocol (MED Associates Inc, St.
Albans, VT,
USA). Animals are individually placed into clear Plexiglas boxes (40.6 x 40.6
x 38.1 cm)
surrounded by multiple bands of photo beams and optical sensors that measure
horizontal and
vertical activity. Mouse movement is detected and recorded for 30 minutes by
breaks within
the beam matrices. Old (e.g. 10 months) drug treated SAMP8 mice have a higher
average
velocity of movement, moving greater distance and a greater number of vertical
movements
than equally old SAMP8 control mice.
(2) Prudence. Elevated Plus maze. Young healthy mice have an aversion to open
spaces.
Mental decline is associated with dis-inhibition and greater comfort/time
spent in open
spaces. The elevated plus maze consists of four arms (two open without walls
and two
enclosed by 15.25 cm high walls) 30 cm long and 5 cm wide in the shape of a
plus. A video
camera mounted overhead on the ceiling linked to video tracking software
(Noldus Etho
Vision) is used to collect behavioural data. This software detects and records
when mice enter
the open or closed arms of the maze and the time spent in each. Mice are
habituated to the
maze for 1 minute before testing by placing them in the centre of the maze and
blocking their
entry to the arms. Dis-inhibition is measured by comparing time spent on open
arms to time
spent on closed arms over a 5 minute testing period. Old (e.g. 10 months) drug
treated
SAMP8 mice have less dis-inhibition than equally old control SAMP8 mice.
(3) Memory, object recognition. Young healthy mice spend more time exploring a
novel
object than a familiar one. Mice are tested in a standard home cage. Phase 1
(Habituation):
Each mouse is placed into the apparatus (no objects present) for two 10 minute
sessions
separated by 1-4 hours to habituate to the testing environment. Phase 2
(Training): Two
identical velcro-backed objects (object "A") are attached into designated
corners of the
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apparatus. The mouse is placed into the apparatus opposite to the objects and
recorded by a
camera for 10 minutes. Phase 3 (Test): One hour after training, the test phase
begins. Only
one of the objects is replaced with a new object (object "B"). The mouse is
placed into the
apparatus opposite to the objects and recorded for 5 minutes. The apparatus is
wiped and
objects cleaned with 70% alcohol to remove odours between mice. "Object
recognition
index" is calculated by dividing the amount of time spent with (touching with
nose or nose
pointing at object and within 0.5 cm of object) object B by the total time
spent with objects A
+ B and multiplied by 100. Old (e.g. 10 months) drug treated SAMP8 mice have a
greater
recognition index than equally old control SAMP8 mice.
(4) Memory, learning, re-learning. Barnes maze: The maze consists of a flat
circular surface
(36" diameter) with 20 equally spaced holes (2" diameter) along the outer
edge. One of the
holes leads to a dark hide box while the other 19 lead to boxes that are too
small to be
entered. The latency to enter the hide box is recorded. The test is conducted
in three phases.
Phase 1 (Training): A hide box is placed under one of the holes. Animals are
placed into an
opaque cylinder in the centre of the maze for 30 seconds to promote spatial
disorientation at
the start of the test. After 30 seconds, the cylinder is removed and the
animal explores the
maze until it finds and enters the hide box. The number of incorrect entries
(nose pokes and
head deflections over any hole that did not have the hide box beneath it) is
scored. If the
mouse fails to enter the box within 3 minutes, it is gently led into the box.
The animal then
remains in the box for an additional 20 seconds before it is removed from the
box and gently
placed into the home cage. Training is repeated three times a day for four
days. The location
of the hide box remains the same during every trial but it is shifted between
subjects to
reduce the potential for unintended intra-maze cues. Phase 2 (Retention): This
phase
measures retention of spatial memory following a delay. After a two day break
from training,
each animal is re-tested for a one day, three-trial session using the same
hide box location as
before. Phase 3 (Reversal): This phase examines memory reversal. On the day
following the
retention phase, a new hide box location is established 180 degrees from the
original location.
The same method as before is used and trials are repeated three times a day
over two
consecutive days. Old (e.g. 10 months) drug treated SAMP8 mice find the hide
box faster,
better retain knowledge of where the hide box is and learn faster a new
location of the hide
box than equally old control SAMP8 mice.
(5) Spatial navigational memory. Two-day water maze. Mouse tracking is
performed using
SMART version 2.0 (Panlab). The water is painted milk-white with nontoxic
paint. A
platform which is visible during training on Day 1 is then submerged just
under the water
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level during testing on Day 2 and mice use spatial cues on the wall around the
pool to
navigate to the platform during testing. During testing on Day 2, the time it
takes each mouse
to find the hidden platform is measured. Old (e.g. 10 months) drug treated
SAMP8 mice find
the hidden platform faster than equally old control SAMP8 mice.
(6) Morris Water Maze [141]. Mouse tracking is performed using SMART version
2.0
(Panlab). The water is painted milk-white with nontoxic paint. 4 trials per
day for 5
consecutive days. For each trial, mice are placed in the pool at 1 of 4 start
locations. The
starting locations are separated by 90 and are termed south, west, north, and
east. Mice start
a trial once from each of the 4 possible start locations on each day. The goal
platform is
positioned 45 cm from the outside wall in the south quadrant of the maze for
all groups. The
latency to find and mount the hidden platform is measured. Swimming speeds are
also
recorded to assess drug-induced motor effects. If the mice fail to find the
platform before 120
seconds expires, they are placed on the platform by the experimenter. Mean
daily latency to
find the goal platform is calculated for each mouse. On day 6, the platform is
removed and
time spent in the platform quadrant determined. The water tank is surrounded
by opaque dark
panels with geometric designs at approximately 30 cm from the edge of the
pool, to serve as
distal cues. Old (e.g. 10 months) drug treated SAMP8 mice swim faster and
reach the hidden
platform faster, and spend longer in the platform quadrant when it is removed,
than equally
old control SAMP8 mice.
(7) Fear conditioning, fear memory, associative learning. A mouse freezes if
it remembers
and associates that environment with an aversive stimulus. Mice are trained on
Day 1 to
associate their environment with an aversive stimulus (a foot shock). The
amount of time
spent freezing in response to the environment is measured on Day 2. Fear
conditioning is
performed in a conditioning chamber (Med Associates) equipped with a grid
floor via which
the foot shock can be administered. Each mouse is placed inside the
conditioning chamber for
180 seconds. A foot shock (2 seconds, 0.4 mA) is delivered 148 seconds after
placement in
the chamber. Twenty-four hours later, context-dependent freezing is measured
during 3
minutes. Time spent freezing is measured using Any-Maze Tm software. To avoid
any
influence of foot shock exposure on further testing, this is the last test
performed of the
battery of tests and all other tests are carried out in tests rooms other than
the fear
conditioning test. Old (e.g. 10 months) drug treated SAMP8 mice spend more
time freezing
in response to the context associated with the aversive stimulus than equally
old control
SAMP8 mice.
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(8) As measured by an assay(s) described in [142], old (10+ months) drug
treated SAMP8
mice have less hearing and/or vision loss (have a lower hearing threshold
and/or a greater
visual contrast sensitivity) than old (10 months) control SAMP8 mice.
(9) As measured by an assay(s) described in [143], old (10+ months) drug
treated SAMP8
mice have greater neuronal plasticity (e.g. greater hippocampal synaptic
plasticity e.g. greater
Long-Term Potentiation [LTP] with excitatory neurons), and less neuronal
degeneration and
reactive astrocytosis, than old (10 months) control SAMP8 mice.
(10) Social preference test (SPT). Assesses sociability and social novelty
preference (i.e.
social recognition memory). The apparatus consists of 3 chambers, a central
chamber (length:
9 cm, width: 18 cm, depth: 20 cm) and two outer chambers (6 cm*18 cm* 20 cm).
The
dividing walls are made of clear Plexiglas, with square passages, 4 cm high
and 4 cm wide.
One circular cage (i.e. mouse enclosure) is placed into each outer chamber.
The mouse
enclosures are 15 cm in height with a diameter of 7 cm and bars spaced 0.5 cm
apart to allow
nose contact between mice but prevent fighting. The chambers and enclosures
are cleaned
with 30% ethanol in-between trials (inter-trial interval of 5 minutes) and
fresh corn cob
bedding is added prior to each test trial. Test animals are isolated for an
hour prior to the start
of testing. During the habituation trial, two mice are placed individually in
the central
chamber and allowed to freely explore the apparatus and the two empty
enclosures for 5
minutes. For the sociability test an unfamiliar adult male mouse is placed in
one of the two
enclosures (i.e. opponent chamber) in a quasi-randomised fashion. Then the
test mouse is
returned to the apparatus and allowed to explore all three chambers for 10
minutes. Finally,
test animals are observed in a 10 minute social recognition test. For this, a
second, unfamiliar
mouse is placed in the previously empty chamber so that the test mouse has the
choice to
explore either the familiar mouse (from the previous trial) or the novel,
unfamiliar mouse.
AnyMazeTm tracking software is used to determine the time spent in the
different chambers,
number of entries and distance travelled by the test mice in each trial. Time
spent sniffing the
opponent is recorded manually (i.e. snout of test mouse within the enclosure
containing the
opponent mouse or <5 mm away from enclosure). Old (e.g. 10 months) drug
treated SAMP8
mice spend more time with the novel individual, as compared to time spent with
the familiar
individual, than equally old control SAMP8 mice.
(11) Olfactory test (i.e. cookie test). Test mice are familiarised with a high
carbohydrate food
(Froot Loops: Kellogg Pty. Ltd., Strawberry Hills, Australia) in their home
cages,
24 hours prior to the test. Consumption is observed by the experimenter to
ensure the novel
food is palatable for the mice. On test day, test mice are habituated for 5
minutes to a large
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opaque cage (47 cm*18 cm*13 cm) containing 2 cm deep bedding. The animal is
removed
from the cage thereafter, and one Froot Loop is buried randomly in the cage
bedding. The
animal is then returned to the cage and given 10 minutes to locate the buried
food. The
latency to find the Froot Loop is recorded. Old (e.g. 10 months) drug treated
SAMP8 mice
will find the Froot Loop faster than equally old control SAMP8 mice.
(12) In treadmill testing, old (e.g. 10 months) drug treated SAMP8 mice have a
faster
maximal running speed, and greater running endurance, than equally old control
SAMP8
mice, whether trained or untrained at treadmill running.
(13) Old (e.g. 10 months) drug treated SAMP8 mice have one or more of better
Blood Brain
Barrier (BBB) homeostasis, less inflammation (e.g. in the brain), less
gliosis, better vascular
function (e.g. in the brain), less Amyloid beta (AO), less tau protein (and/or
less
hyperphosphorylation of tau protein), lower levels of Vascular Cell Adhesion
Molecule 1
(VCAM-1, a protein associated with vascular endothelium inflammation), lower
levels of
endogenous immunoglobulin G (IgG, high levels observed in old mice as
consequence of
disrupted BBB permeability), less glial fibrillary acidic protein (GFAP)
expression, increased
brain Docosahexaenoic Acid [DHA] (possibly because of less oxidation of DHA,
DHA is the
primary structural fatty acid in the human brain and has been linked to
cognitive
performance. Low plasma levels of DHA are associated with cognitive decline in
elderly and
Alzheimer's disease patients, higher DHA intake and plasma levels inversely
correlate with
Alzheimer's disease risk, DHA supplementation in aged animals enhances
learning and
memory [125]), increased brain glutamate levels (brain [glutamate] decrease
with age [122]
and low [glutamate] has been observed with Alzheimer's disease [123, 124])
and/or a lesser
pro-oxidant status in the brain than equally old control SAMP8 mice.
(14) Old (e.g. 10 months) drug treated SAMP8 mice have less aging (are more
similar to
young SAMP8 mice), at one or more of the
cognitive/movement/anatomical/physiological/electrophysiological/cellular
(e.g. number of
senescent cells [126])/biochemical/neurochemical/protein/protein modification
(e.g.
carbamylation [127])/oxidation e.g. [128,
129]/metabolite/metabolic/epigenetic/histone
loss/histone modification/telomere length/gene expression/DNA/DNA modification
(e.g.
DNA methylation)/RNA levels, than equally old control SAMP8 mice, for example
as
reported using one or more of the assays described in [130, 131, 132, 133,
134, 135. 136,
137, 138] or some other aging assay(s) of the art e.g. as described in the
literature e.g.
using/leveraging an aging/mortality biomarker(s) reported in the database:
http://mortalitypredictors.org/ [139].
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(15) Transcriptional drift is an age-associated loss of coordination among
groups of genes
[144, 145]. Aging causes genes within functional groups to change expression
in opposing
directions, which cause a transcriptome-wide loss in mRNA stoichiometry and
loss of co-
expression patterns in aging animals, as compared to young animals. Observing
hippocampal
gene expression data, old (e.g. 10 months) drug treated SAMP8 mice have less
transcriptional
drift than equally old control SAMP8 mice i.e. old (e.g. 10 months) drug
treated SAMP8
mice have a transcriptome (e.g. hippocampal transcriptome) more similar to
young SAMP8
mice than equally old control SAMP8 mice. Metabolomic/metabolic drift is an
age-associated
change in the relative/absolute amounts of metabolite(s) e.g. reduced [NAD]
[146, 138],
increased AMP/ATP etc. [147]. Old (e.g. 10 months) drug treated SAMP8 mice
have a (e.g.
plasma and/or brain [e.g. hippocampal]) metabolome more similar to young SAMP8
mice
than equally old control SAMP8 mice i.e. old (e.g. 10 months) drug treated
SAMP8 mice
have less (e.g. plasma and/or brain [e.g. hippocampal]) metabolomic/metabolic
drift than
equally old control SAMP8 mice. Optionally metabolome analysis is performed
using
Precision MetabolomicsTM (Metabolon Inc., Morrisville, NC, USA).
HYPERMETABOLISM
An invention embodiment is a method in which a subject takes or is
administered an effective
amount of a compound(s) of this invention, for example a compound of Formula
(I), (II),
(III), (IV), (V) or (VII) or another compound that selectively inhibits FIR'
ATP hydrolysis, or
a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
optionally in co-
therapy with one or more of an anti-thyroid drug(s) (illustrating, without
restriction,
carbimazole, methimazole, propylthiouracil/PTU, potassium perchlorate),
radioiodine, beta
blocker(s) (illustrating, without restriction, propranolol, metoprolol),
surgery
(thyroidectomy), to treat/ameliorate/prevent/combat one or more of
hypermetabolism, heat
intolerance, thyroidal hypermetabolism, non-thyroidal hypermetabolism e.g.
Luft's disease.
COMPOUNDS OF THIS INVENTION ARE ANXIOLYTICS, ANTICONVULSANTS,
ANTIPSYCHOTICS, ANTIDEPRESSANTS, ANTIEMETICS, ANALGESICS,
SEDATIVES, HYPNOTICS AND ANTIHISTAMINES
As reported in the legend of Figure 23 of this disclosure, when mice were
administered
Compound 6b of this invention they exhibited hypoactivity, the duration of
which correlated
with their drop in rectal temperature, which correlated with the administered
dosage of 6b,
wherein greater 6b dose caused greater rectal temperature drop and greater
hypoactivity.
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CA 3050553 2019-07-25
Hypoactivity of mouse, after being administered compound 6b, was because 6b
caused the
mouse's body temperature to drop towards its ambient temperature (22 C). When
ambient
equals optimal body temperature (37 C), 6b doesn't/can't reduce body
temperature and
doesn't cause hypoactivity. In some invention embodiments, the
hypoactivity/sedation aspect
to a compound(s) of this invention is utilized for therapy. The intersection
between drug dose
and ambient temperature dictates how much the body temperature falls and
thence the depth
of the sedation. Larger drug dose, and/or lower ambient temperature, causes
deeper sedation
(e.g. useful for inducing anaesthesia, pre-anaesthesia, post-anaesthesia,
hypoesthesia,
sedation, coma, tranquilization, behavioural submission, muscle relaxation
and/or
treating/ameliorating/preventing/combating insomnia, fatal insomnia, sleep
onset latency,
delayed sleep phase disorder, exploding head syndrome, parasomnia, sleep-
maintenance
insomnia, sleep disorder etc.; repeated/continuous administration, e.g.
repeated i.v. injections
or continuous i.v. infusion, of a compound(s) of this invention can sedate a
subject for an
extended period of time). Smaller drug dose, and/or higher ambient
temperature, causes
lighter sedation (e.g. useful for anti-anxiety, anti-depression, hyperactivity
etc.). At
ambient>optimal body temperature (37 C), no sedation occurs. Increasing
sedation occurs
with lower body temperature because action potential characteristics are
temperature
dependent. In some invention embodiments, deeper sedation is conveyed by a
larger body
temperature drop (body temperature<34 C) and in other invention embodiments a
slight
calming sedation, optionally imperceptible to the subject, is conveyed by a
smaller body
temperature drop, optionally less than 1 C, optionally less than 0.5 C. To
repeat, the
intersection between dose and ambient temperature dictates the magnitude of
body
temperature drop, wherein this can be zero, even at high drug dose, when
ambient
temperature >37 C. So many diseases/disorders are because of too
much/inappropriate/undesired signals/activity/electrical activity in the
nervous system,
wherein a compound(s) of this invention can decrease nervous system activity
by a tunable
degree (reduction amplitude set by intersection of drug dose with ambient
temperature) and
so can treat/ameliorate/prevent/combat an incredibly large number of
diseases/disorders.
Fundamental action equals broad applicability. Drug action against a
fundamental
physiological parameter (body temperature), which dictates a further
fundamental
physiological parameter (action potential characteristic(s): firing
threshold/conduction
velocity/firing frequency etc.), yields broad therapeutic application. For
example, it raises the
stimulus threshold for an epileptic seizure, which decreases the frequency of
epileptic
seizures in a subject. For example, it raises the stimulus threshold for
ejaculation, therefore
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delaying ejaculation during sex, therefore assisting a subject with premature
ejactulation. For
example, it raises the threshold for pain perception and so any pain is
reduced in magnitude.
For example, in the brain, it raises the threshold for a "behaviour suggestion
signal" to be
selected for action, thence reducing the occurrence of unwanted behaviour
(e.g. in Tourette's
syndrome).
Reduced temperature decreases action potential (AP) conduction velocity (Q10 =
¨1.7, so AP
velocity is ¨10% less at 35 than 37 C [148, 149]), decreases AP frequency (30%
decrease in
spike rate with 2 C decrease in temperature [150]), increases AP firing
threshold (AP firing
threshold vs. temperature is U shaped because threshold increases as move away
from an
optimum temperature [15 I, 152]) and decreases neural circuit activity in vivo
[153].
An embodiment of this invention is to administer to a subject compound 6b,
and/or 7b, and/or
another compound(s) of this invention, i.e. a compound that preferentially
inhibits FIR, ATP
hydrolysis over FiFo ATP synthesis, for example a compound(s) of Formula (I),
(II), (III),
(IV), (V) or (VII) or a pharmaceutically-acceptable salt, solvate, hydrate or
prodrug thereof,
to treat/ameliorate/prevent/combat hyperactivity, hypersensitivity, Premature
ejaculation,
hyperreflexia, Autonomic dysreflexia (AD), Hyperventilation syndrome, brain
hyperactivity,
overly sensitive sensory system, pathological crying and/or laughing,
Pseudobulbar affect
(PBA, emotional lability), photophobia, phonophobia, temperature-sensitive,
pressure-
sensitive, brain hyperexcitability, overstimulation, intrusive thought(s),
Perseveration,
sensory overload, disorganized thinking, fantasy prone personality,
malapdative
daydreaming, dissociation, hyperkinetic disorder, agitation, Psychomotor
agitation,
restlessness, difficulty controlling behaviour, disruptive behaviour disorder,
Emotional and
.. behavioral disorder, pervasive developmental disorder, Overactive disorder
associated with
mental retardation and stereotyped movements, attention-deficit disorder,
Attention Deficit
Hyperactivity Disorder (ADHD), adult attention-deficit hyperactivity disorder,
severe
behavioral problem(s) in children (e.g., to illustrate and not restrict,
combativeness and/or
explosive hyperexcitable behavior {out of proportion to immediate
provocation[s]},
hyperactive children who show excessive motor activity with accompanying
conduct
disorders consisting of one or more of: impulsivity, difficulty sustaining
attention,
aggressivity, mood lability, poor frustration tolerance), Premenstrual
dysphoric disorder
(PMDD), premenstrual syndrome (PMS), impulsiveness, impulsivity, impulse
control
disorder, lack of self-control, hysteria, histrionic personality disorder,
attention difficulty,
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CA 3050553 2019-07-25
inattention, poor attention control, anxiety, paranoid anxiety, Paranoid
personality disorder,
distress, dysphoria, Adjustment disorder, separation anxiety, anxiety
disorder, depressive
anxiety, agitated depression, treatment-resistant depression, Generalized
anxiety disorder,
social anxiety disorder, stranger anxiety, separation anxiety (e.g. in dogs
left at home),
separation anxiety disorder, Mixed anxiety-depressive disorder, depression
(all forms, all
severities), restlessness/apprehension/anxiety before surgery. hypochondria,
panic disorder,
panic attack, emotional outburst, emotional instability, Intermittent
explosive disorder,
unreasonable/unwarranted anger/aggression, hyper-aggression, hostility, rage,
poor temper
control, self-hatred, poor attentional control, worry, irritability, neuroses,
somatization
.. disorder, somatic symptom disorder, pain disorder, psychological pain,
psychogenic pain,
psychogenic facial pain, Atypical odontalgia (AO), burning mouth syndrome,
throbbing,
toothache/pulpitis/dental pain, chronic lower back pain, negative emotion,
persistent/enduring
negative emotion, body dysmorphic disorder, factitious disorder, illness
anxiety disorder,
unwarrented fight-or-flight response, stress, emotional stress, emotional
dysregulation,
distress, psychological stress, acute stress, chronic stress, acute stress
reaction, combat stress
reaction, traumatic grief, grief, grief after death of loved one, Prolonged
grief disorder (PGD),
heartbreak, guilt, shame, remorse, emotional pain, Algopsychalia, psychalgia,
suffering,
emotional trauma, psychological trauma, broken heart, Post Traumatic Stress
Disorder
(PTSD), Complex post-traumatic stress disorder (C-PTSD), hypervigilance,
sympathetic
.. hyperactivity, inability or impaired ability to relax, flashbacks,
dysphoric hyperarousal,
agoraphobia, insomnia, fatal insomnia, mood disorder, irrational/unwarrented
fear/terror,
phobia, social phobia, Cancerophobia, thunderstorm/firework phobia,
hypersexuality,
hypersexual disorder, depression, clinical depression, unipolar depression,
bipolar disorder,
Bipolar 1, Bipolar II, Bipolar disorder not otherwise specified (Bipolar NOS),
cyclothymia,
cyclothymic disorder, mixed affective state, atypical depression, melancholic
depression,
postpartum depression, double depression, seasonal affective disorder, mania,
manic episode,
hypomania, increase in energy of psychomotor activity, delirium, excited
delirium, major
depressive disorder, minor depressive disorder, recurrent brief depression,
Depressive
Disorder Not Otherwise Specified (DD-NOS), major depressive episode,
persistent
depressive disorder (PDD), dysthymia, dysthymic disorder, absence of euthymia,
manic
thoughts, racing thoughts, thought disorder, disordered thinking, reduced
ability to plan and
execute tasks, paranoia, hallucination (including, without limitation, visual,
auditory,
olfactory, gustatory, tactile, proprioceptive, equilibrioceptive, nociceptive,
thermoceptive,
chronoceptive), Charles Bonnet syndrome, delusion, persecutory delusion,
hearing voices,
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CA 3050553 2019-07-25
homicidal/criminal ideation/tendency/thoughts, suicidal
ideation/tendancy/thoughts, self-
injury, non-suicidal self-injury, violence, attacking others, negative mood
swing, personality
disorder, Borderline personality disorder, Narcissistic personality disorder,
malignant
narcissism, dissociative disorder, dissociative identity disorder (DID),
Psychosis, acute
psychosis, chronic psychosis, psychosis spectrum disorder, manifestations of
psychotic
disorders, behavioral complications of mental retardation, stimulant
psychosis, psych'otic
depression, hallucinogen persisting perception disorder, Psychoactive
substance-related
disorder, amphetamine-induced psychosis, brief psychotic disorder, Brief
reactive psychosis,
Menstrual psychosis, postpartum psychosis, Psychotic disorder, Psychopathy,
chronic
hallucinatory psychosis, manifestation(s) of psychotic disorder,
neurotic/reactive/endogenous/involutional/psychotic depression/depressive
disorder
(optionally accompanied by anxiety or agitation), depressive neurosis,
delusional depression,
psychotic aggression, psychiatric symptoms of dementia, AIDS delirium,
Supersensitivity
psychosis, Tardive psychosis, Tardive dysmentia, Depersonalization disorder,
out-of-body
experience, Sociopathy, Schizophrenia, Paranoid schizophrenia, disorganized-
type
schizophrenia, simple-type schizophrenia, pseudoneurotic schizophrenia,
prodromal
schizophrenia, schizoaffective disorder, bipolar type schizoaffective
disorder, depressive type
schizoaffective disorder, schizoaffective psychosis, Schizotypal personality
disorder,
schizophreniform disorder, Delusional parasitosis, formication, paresthesias,
Acroparesthesia,
tinnitus, delusional disorder, delusional jealousy, inappropriate behaviour,
behavioural
disorder, antisocial personality disorder, Oppositional defiant disorder
(ODD), conduct
disorder (CD), Disruptive mood dysregulation disorder (DMDD), sadistic
personality
disorder, poor inhibitory control, kleptomania, Pyromania, trichotillomania,
dermatillomania,
pathological/problem gambling, dyskinesia, tardive dyskinesia, paroxysmal
dyskinesia,
Paroxysmal kinesigenic dyskinesia, Paroxysmal nonkinesigenic dyskinesia,
Paroxysmal
exercise-induced dystonia, Hemiballismus, tic disorder, tremor, clonus,
Tourette's syndrome,
coprolalia, copropraxia, Echophenomena, Echopraxia, Echolalia, Palilalia,
stuttering/stammering, stereotypy, punding, Self-stimulatory behaviour
(stimming),
Stereotypic movement disorder (SMD), synesthesia, obsession,
Obsessive¨compulsive
disorder (OCD), obsessive¨compulsive personality disorder, anankastic
personality disorder,
relationship obsessive¨compulsive disorder (ROCD), Scrupulosity, Primarily
obsessional
obsessive compulsive disorder, sexual obsession, Akathisia (including, without
limitation,
chronic, acute, Pseudoakathisia, Tardive akathisia, withdrawal or "rebound"
akathisia),
Restless legs syndrome, motor restlessness, periodic limb movement disorder
(PLMD),
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CA 3050553 2019-07-25
periodic limb movements in sleep (PLMS), Sydenham's chorea, chorea, dystonia,
Hypnic
jerk, twitching, spasms, Tetanus, tetanus muscle spasms, tetanized state,
Myoclonus,
myoclonic seizure, Action myoclonus, Palatal myoclonus, Middle ear myoclonus,
Spinal
myoclonus, Stimulus-sensitive myoclonus, Sleep myoclonus, Cortical reflex
myoclonus,
Essential myoclonus, myoclonic epilepsy, Juvenile myoclonic epilepsy,
Progressive
myoclonus epilepsy (PME, including, without limitation, Dentatorubral-
pallidoluysian
atrophy, Unverricht-Lundborg disease, MERRF syndrome, Lafora disease),
Reticular reflex
myoclonus, Myoclonic epilepsy, diaphragmatic flutter, automatism, status
epilepticus,
Epilepsia partialis continua, Complex partial status epilepticus, epilepsy,
epileptic seizure,
simple partial seizure, complex partial seizure, generalized epilepsy,
generalized seizure
(including, without limitation, tonic-clonic, tonic, clonic, myoclonic,
absence (including
typical absence and atypical absence), atonic seizure), focal epilepsy, focal
seizure,
focal/partial seizure (including, without limitation, Simple partial seizure
and Complex partial
seizure), focal aware seizure, focal impaired awareness seizure, generalised
epilepsy,
temporal lobe epilepsy (including, without restriction, mesial temporal lobe
epilepsy
{MTLE} and lateral temporal lobe epilepsy {LTLE}), Frontal lobe epilepsy,
Rolandic
epilepsy, Nocturnal epilepsy, Nocturnal frontal lobe epilepsy, Autosomal
dominant nocturnal
frontal lobe epilepsy (ADNFLE), Generalized epilepsy with febrile seizures
plus (GEFS+),
Panayiotopoulos syndrome, epileptic fit, reflex epilepsy, reflex seizure,
absence seizure
(including, without limitation, childhood absence epilepsy, epilepsy with
myoclonic
absences, juvenile absence epilepsy, juvenile myoclonic epilepsy, Jeavons
syndrome {eyelid
myoclonia with absences}, genetic generalised epilepsy with phantom absences),
complex
partial seizure, atonic seizure, generalized tonic-clonic seizure,
tonic¨clonic seizure, extrinsic
stimulus epilepsy, intrinsic stimulus epilepsy, photosensitive epilepsy,
musicogenic epilepsy,
thinking epilepsy, eating epilepsy, seizure(s), Febrile seizure, nerve agent
induced seizure,
Dravet syndrome (sometimes modest hyperthermic stressors like physical
exertion or a hot
bath can provoke seizures in affected individuals), acute symptomatic seizure,
seizure-related
disorder, drug related seizure, paroxysmal depolarizing shift, Ohtahara
syndrome, Epilepsy in
females with mental retardation, Rasmussen's encephalitis, Epilepsy syndrome,
benign
rolandic epilepsy, childhood absence epilepsy, absence epilepsy, juvenile
myoclonic
epilepsy, epileptic encephalopathies, Lennox¨Gastaut syndrome, West syndrome
(Epileptic
spasms), Doose syndrome (Myoclonic astatic epilepsy, MAE), Lennox-Gestaut
syndrome,
pseudo-Lennox Gastaut syndrome (atypical benign partial epilepsy), Benign
familial neonatal
epilepsy, Benign occipital epilepsy of childhood, familial neonatal
convulsions, Febrile
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CA 3050553 2019-07-25
infection-related epilepsy syndrome, interictal dysphoric disorder, euphoria
sclerotic,
psychogenic non-epileptic seizure, non-epileptic seizure, gelastic seizure,
convulsion(s),
migraine, status migrainosus, tension headache, headache, Hypnic headache,
hiccups,
intractable hiccups, thumps in equines, Postural orthostatic tachycardia
syndrome (POTS),
Pheochromocytoma, agony, pain, chronic pain, acute pain, pain due to
disease/injury,
neuropathic pain, fibromyalgia, postherpetic neuralgia, phantom pain, referred
pain, back
pain, lower back pain, pelvic pain, cancer associated pain, chemotherapy
associated pain,
Diabetic neuropathy, small fiber peripheral neuropathy, Mononeuritis
multiplex,
Wartenberg's migratory sensory neuropathy, Breakthrough pain, idiopathic pain,
polyneuropathy, Neuritis, Mononeuropathy, Polyradiculoneuropathy, Radial
neuropathy,
neuropathy, visceral pain, Burning feet syndrome, Tarsal tunnel syndrome,
Carpal tunnel
syndrome, Guyon's canal syndrome, Cubital Tunnel Syndrome, Repetitive strain
injury,
Sciatica, Neurofibromatosis, Ulnar nerve entrapment, Erythromelalgia,
Paroxysmal extreme
pain disorder, Proctalgia fugax, dysesthesia, scalp dysesthesia, dysesthetic
burning,
hyperesthesia, hyperalgesia, Opioid-induced hyperalgesia, hyperpathia,
allodynia, pain
response from stimuli which do not normally provoke pain, Complex regional
pain syndrome
(said to be most painful condition known to man), Radiculopathy, neuralgia
(including,
without restriction, intercostal neuralgia, trigeminal neuralgia, atypical
trigeminal neuralgia,
glossopharyngeal neuralgia, occipital neuralgia, postherpetic neuralgia),
ciguatera poisoning,
irritable bowel syndrome (IBS), interstitial cystitis (IC), temporomandibular
joint disorder,
acute intermittent porphyria, Porphyria, Acute porphyria (including, without
limitation, acute
intermittent porphyria {AIP}, variegate porphyria {VP}, aminolevulinic acid
dehydratase
deficiency porphyria {ALAI)}, hereditary coproporphyria {HCP}, drug induced),
Chronic
porphyria (including, without limitation, X-linked dominant protoporphyria
{XLDPP},
congenital erythropoietic porphyria {CEP}, porphyria cutanea tarda {PCT}, and
erythropoietic protoporphyria {EPP}), cutaneous porphyria, Porphyria cutanea
tarda, allergy,
allergic reaction, anaphylaxis, anaphylactic shock, hives, asthma, allergic
rhinitis, rhinitis,
urticaria, contact dermatitis, cough, sore throat, esophageal reflux disease,
heartburn, chest
pain, Esophageal motility disorder, Nutcracker esophagus, diseases involving
gastrointestinal
motility, Peptic ulcer disease, esophageal spasm, angina, itchiness, Pruritus,
severe pruritus,
Prurigo, Pruritic skin condition, chronic itch, eczema, pruritus in eczema,
neuropathic itch,
neurogenic itch, itchiness due to atopic dermatitis and/or lichen simplex
chronicus, peripheral
sensitization, central sensitization, sensory perception of absent stimuli,
too much sensory
stimulation, sensory stimulation brings discomfort, Neuromyotonia, Peripheral
nerve
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hyperexcitability, Morvan's syndrome, Benign fasciculation syndrome, Cramp
fasciculation
syndrome, hyperhidrosis, undifferentiated somatoform disorder, somatoform
disorder,
somatic symptom disorder, conversion disorder, functional neurological symptom
disorder,
severe nausea and/or vomiting, severe nausea/emesis, Chemotherapy-induced
peripheral
neuropathy, chemotherapy-induced nausea and vomiting (CINV), radiation therapy-
induced
nausea and vomiting (RINV), postnarcotic nausea, hyperemesis gravidarum,
morning
sickness, Cyclic vomiting syndrome, retching/dry heaving, Urinary
incontinence, enuresis,
nocturnal enuresis, tail chasing, reduce urine spraying/marking behavior,
benzodiazepine
withdrawal syndrome, barbituate withdrawal syndrome, Neuroleptic
discontinuation
syndrome, drug withdrawal discomfort/pain/symptoms, drug use disorder, alcohol
use
disorder, opoid use disorder, amphetamine use disorder, cocaine use disorder,
alcohol
withdrawal syndrome/symptoms, delirium tremens, opoid withdrawal
sydrome/symptoms,
drug craving, drug addiction, drug dependence, polysubstance dependence, drug
overdose,
smoking, tobacco (nicotine) addition, tobacco (nicotine) withdrawal symptoms,
alcoholism,
addiction, opoid addiction, cocaine/crack addiction, addictive behaviour,
addictive
personality, behavioural addiction, internet/computer/computer game/social
media/media
addiction, exercise addiction, compulsive behaviour (e.g. {non-limiting}
checking, counting,
washing, repeating), anti-social behaviour, criminality, sexual compulsion,
impulsive sexual
behaviour, compulsive buying, gambling addiction, sex related addiction,
sexual urge,
hunger, eating desire/compulsion, eating disorder, polyphagia, overeating,
binge eating
disorder, compulsive overeating, insatiable/excessive appetite, bulimia
nervosa, anorexia
nervosa, substance abuse, substance-induced delirium, substance-induced
psychosis,
substance-induced mood disorder, drug overdose, vertigo, motion sickness,
seasickness,
mental/nervous breakdown, Autism spectrum disorder, neurological disorder,
cognitive
disorder, mental disorder, mental health disorder, mental health condition
involving impaired
or altered neural plasticity, mood disorder, mental disorder disclosed in
Diagnostic and
Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) or a later
edition, a
mental/behavioural disorder disclosed by the International Classification of
Diseases (ICD) in
ICD-10 Chapter V: Mental and behavioural disorders (World Health Organisation,
WHO).
Because a compound(s) of this invention can cause sedation (when
ambient<optimal body
temperature {37 C}) and slow aging, this juxtaposition makes a compound(s) of
this
invention useful for inducing hibernation/artificial
hibernation/torpor/synthetic
torpor/suspended animation, optionally used on a long journey, optionally
during spaceflight,
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optionally on a journey to Mars (projected duration with present technology is
¨18 months
transit time for round trip). Moreover because the compound(s) reduces food,
power (e.g.
reduced lighting/heating), living space and 02 requirements in a subject, this
makes the
spacecraft load lighter, and the induced lower respiration rate means
slower/shallower
breathing, less 02 in the body, which reduces the damaging effect of ionizing
radiation
(significant in space), moreover permitting a lower 02 concentration outside
the body, which
reduces ionizing radiation damage to the outside of the body, and the smaller
living space
afforded by the sedation permits greater radiation shielding per unit living
space and the
induced hypometabolism reduces the rate of muscle and bone atrophy (reduces
spaceflight
osteopenia) and other negative health effects (e.g. sleep disturbance) of
microgravity, and the
sedation side-steps the anticipated worrying problem of interpersonal
friction(s) during long
confined spaceflight (Cosmonaut Valery Ryumin's autobiography: "If you want to
instigate
the art of manslaughter just shut two men up in a eighteen by twenty-foot
cabin for a month.
Human nature won't stand it."). Optionally the drug can be administered by
continuous
intravenous infusion, wherein optionally respiratory substrates, nutrients,
fluids etc. can be
administered similarly (e.g. using parenteral nutrition). If there is a
job/emergency that needs
to be attended to in the spacecraft (and/or the subject is to
eat/wash/administrate themselves
etc.) the hibernation is paused by raising the ambient temperature of the
subject to 37 C.
Afterwards, assuming the subject still has sufficient compound(s) in their
system, the
hibernation can be induced again by lowering the ambient temperature. During
spaceflight,
the use of a compound(s) of this invention combats many of the problems
identified for space
exploration/travel by NASA Report No. IG-16-003 ("NASA's efforts to manage
health and
human performance risks for space exploration", October 29 2015, audit
conducted by Office
of Inspector General).
An aspect of this invention is to use a compound(s) of this invention to
sedate a subject
undergoing treatment, and/or a course of treatment, for a
pathology/disease/disorder/dysfunction/unwanted characteristic(s) of the
subject. For non-
limiting example, for a subject undergoing drug (e.g. opoid) withdrawal,
wherein a
compound(s) of this invention is used to sedate the subject during their drug
withdrawal
phase so they don't suffer the, typically horrific (why many drug users can't
get off drugs),
withdrawal symptoms such as pain, nausea, craving etc., which are worst in the
first few days
of withdrawal, a common time of drug relapse.
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Because a compound(s) of this invention can cause sedation (when
ambient<optimal body
temperature{37 C}), slow aging and exert anti-cancer activity, these
attributes make a
compound(s) of this invention useful for a subject undergoing anti-cancer
treatment,
optionally during a hospital stay, wherein more than the time the subject
loses sedated is
returned to them by a longer [life/health]span. When the cancer patient has a
visitor the
sedation can be paused by raising the ambient temperature to 37 C (e.g. by
transferring the
patient's bed trolly into a visitors area/room maintained at this
temperature). If a light
sedation (small body temperature drop, cancer patient remains conscious but
calmer, cancer
patient can go about their normal life) is instead chosen for a cancer
subject, there is a useful
juxtaposition in the compound(s) anti-cancer and anxiolytic and/or
antidepressant effects,
because many cancer patients are anxious/depressed, and there is benefit to
the compound's
analgesic and/or antiemetic effects also, if radio/chemo-therapy is used in co-
therapy, because
radio/chemo-therapy typically causes cancer patients pain and nausea/vomiting,
often
extreme. Optionally a compound(s) of this invention is taken before the
subject wishes to
sleep, for example at night, and so any perceptible sedation, should it occur
with the dose
taken at that ambient temperature, is then virtuous rather than limiting to
normal life.
SURGERY
An embodiment of this invention is a method in which a subject takes or is
administered
and/or has a plasma blood level of an effective amount of a compound(s) of
this invention,
for example a compound of Formula (I), (II), (III), (IV), (V), (VI) or (VII)
or another
compound that selectively reduces FIR) ATP hydrolysis, or a pharmaceutically-
acceptable
salt, solvate, hydrate or prodrug thereof, and the subject is administered an
analgesic(s)
and/or an anti-nausea medication(s) and/or an anxiolytic and/or antidepressant
and/or a local
and/or a general anaesthetic (examples well known to those of the art), and/or
optionally
another drug commonly used in surgery, such as an anti-anxiety/sedative drug
used before
general anaesthesia or after surgery (examples are well known to those of the
art), optionally
to treat/ameliorate/prevent/combat cancer, optionally wherein the subject
undergoes surgery
to remove a tumour(s).
EMERGENCY CONTRACEPTIVES
Adult bodies don't contain embryonic stem (ES) cells. ES cells are in the
inner cell mass of
the blastocyst, which forms ¨5 days post-fertilization, and exist temporarily
because they
soon differentiate into other cell types, without ES cell characteristics. So,
a compound(s) that
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specifically kills ES cells will have utility as an emergency contraceptive,
taken after
unprotected sex for example, with a later window of effectiveness than present
emergency
contraceptives. Furthermore, it will have utility as a contraceptive, which
can be administered
instead of, or in combination with, another contraceptive(s) such as the
combined oral
contraceptive pill, wherein this term and the range of compositions it can
refer to are well
known to those of the art.
Cancer cell metabolism is similar to that of ES cells. Both can proliferate
rapidly, forever
(without limit, immortal). They share gene expression fingerprints [54]. ES
cells also have a
hyperpolarised Tim [55], employ aerobic glycolysis some or all of the time
[56] and tend to
respond to ROS damage by apoptosis rather than repair [57].
Fi Fo ATP hydrolysis inhibitors have anti-cancer activity, as disclosed by
this invention, and,
also by this invention, anti-ES cell activity. An embodiment of this invention
is a
compound(s) that inhibits FiFo ATP hydrolysis, for example a compound(s) of
Formula (I-V,
VII), administered or self-administered to a subject, for use in
preventing/ending their
pregnancy/conception, optionally co-administered (optionally in a
pharmaceutical
composition) with another compound(s) or combination of compounds with this
use, many of
which are known to those of the art .e.g. progestin, antiprogestin, estrogen
etc. This use could
be after unprotected sex for example. In another embodiment, this use is
restricted to the time
during which ES cells exist in embryogenesis, which is early.
An embodiment of this invention is an FIR ATP hydrolysis inhibitor(s) in use
as an anti-
cancer medicine, or some other therapeutic use in a subject, wherein the
compound(s) is
distributed, sold and/or administered with a verbal and/or written warning,
optionally in a
paper insert in a packet containing the compound(s), that it should not be
administered to a
woman in the early days and/or weeks of pregnancy and/or in a woman trying to
get
pregnant.
COMPOUNDS OF THIS INVENTION ARE ANTI-INFLAMMATORIES
An embodiment of this invention is a method of using a therapeutically
effective amount of at
least one compound of this disclosure, which inhibits FIFO ATP hydrolysis, as
an
immunosuppressant and/or anti-inflammatory therapeutic. This therapeutic
opportunity exists
because if TN collapses in a cell, apoptosis ensues [59], and activated
macrophages, unlike
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resting macrophages, singly use and thence completely rely upon ATP synthase
in its reverse
mode, hydrolysing ATP, to maintain Tim [154]. This is because activated
macrophages
produce nitric oxide (NO), which switches down/off their OXPHOS (NO increases
the Km of
Complex IV for 02). Compounds of the present invention inhibit FiFo ATP
hydrolysis and
selectively collapse TIM in activated macrophages, selectively killing them
(optionally
administered in co-therapy with an uncoupler(s), to erode Tim as the FiFo ATP
hydrolysis
inhibitor(s) blocks the only means activated macrophages can counter this
erosion; thence
Tim collapses at lower dose of FIFO ATP hydrolysis inhibitor). Thus a
therapeutically
effective amount of at least one compound of this invention, an FIFO ATP
hydrolysis
inhibitor, for example a compound of Formula (I), (II), (III), (IV), (V) or
(VII) or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof
(optionally in co-
therapy with an uncoupler(s), optionally administered in the same
pharmaceutical
composition and/or administered/packaged/sold together; in some embodiments
the
therapeutic utility of the FIN ATP hydrolysis inhibitor(s) synergizes with
[potentiates] that of
the uncoupler(s); optionally the same compound is both an FIR ATP hydrolysis
and
uncoupler e.g. BMS-199264), attenuate the activated macrophage (or similar
cell type)
component to inflammation, and its pathologies, and
treats/ameliorates/prevents/combats any
disease or disorder associated with the undesirable activation and/or activity
and/or number
of macrophages/pancreatic islet macrophages/Langerhans cells/dendritic
cells/monocytes/histiocytes/Hofbauer cells/Kupffer
cells/phagocytes/microglia/epithelioid
cells/osteoclasts/macrophage like cells/cells of the mononuclear phagocyte
system, and/or
any cell type(s) of the innate immune system and/or of the monocyte lineage,
especially
inducible nitric oxide synthase (iNOS) and/or iN0S2 expressing and/or NO
producing cells
(e.g. monocyte-derived inflammatory dendritic cells), and/or immune or
inflammation
diseases/disorders/pathologies including, but not limited to, acute
inflammation, chronic
inflammation, systemic inflammation, inflammation because of infection or
foreign bodies or
injury or chemical or toxin or drug or stress or frostbite or burn or ionising
radiation,
inflammatory/neuroinflammatory diseases/disorders/syndromes, Macrophage
Activation
Syndrome (MAS), autoinflammatory diseases/disorders/syndromes, age-related
chronic
inflammatory diseases ("inflammaging"), autoimmune
diseases/disorders/syndromes,
diseases/disorders of the innate immune system, sore throat, sore throat
associated with cold
or flu or fever, high-intensity exercise associated inflammation, ulcerative
colitis,
inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), rheumatoid
arthritis,
osteoarthritis, inflammatory osteoarthritis, psoriatic arthritis, atopic
dermatitis, allergic airway
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inflammation, asthma, inflammation associated depression, neuroinflammation,
neuropathic
pain, exercise-induced acute inflammation, atherosclerosis, allergy, hay
fever, anaphylaxis,
inflammatory myopathies, drug-induced inflammation, systemic inflammatory
response
syndrome, sepsis-related multiple organ dysfunction/multiple organ failure,
microbial
infection, acute brain/lung/hepatic/renal injuries, lung inflammation, acute
lung injury
(ARDS), acne vulgaris, celiac disease, celiac sprue, chronic prostatitis,
colitis, autoimmune
hemolytic anemia, diverticulitis, glomerulonephritis, proliferative
glomerulonephritis,
membranous nephropathy, minimal change nephrotic syndrome, hidradenitis
suppurativa,
hypersensitivities, interstitial cystitis, Mast Cell Activation Syndrome,
mastocytosis, otitis,
pelvic inflammatory disease (PID), endometritis, reperfusion injury, rheumatic
fever, rhinitis,
sarcoidosis, transplant rejection, parasitosis, eosinophilia, type III
hypersensitivity, ischaemia,
chronic peptic ulcer, tuberculosis, Crohn's disease, hepatitis, chronic active
hepatitis, immune
hepatitis, alcoholic hepatitis, chronic viral hepatitis, ankylosing
spondylitis, diverticulitis,
fibromyalgia, systemic lupus erythematous (SLE), Alzheimer's disease,
Parkinson's disease,
neurodegenerative disease, cardiovascular disease, chronic obstructive
pulmonary disease,
bronchitis, acute bronchitis, bronchiectasis, bronchopneumonia, obliterative
bronchiolitis,
appendicitis, acute appendicitis, bursitis, cystitis, dermatitis,
encephalitis, HIV encephalitis,
gingivitis, meningitis, infective meningitis, myelitis, nephritis, neuritis,
periodontitis, chronic
periodontitis, phlebitis, prostatitis, RSD/CRPS, rhinitis, sinusitis, chronic
sinusitis, tendonitis,
testiculitis, tonsillitis, urethritis, vasculitis, respiratory
bronchiolitis¨associated interstitial
lung disease and desquamative interstitial pneumonia, pneumonia, interstitial
lung disease,
Ltifgren syndrome, Heerfordt syndrome, monocytosis, liver fibrosis,
steatohepatitis,
nonalcoholic steatohepatitis, silicosis, histiocytoses, Langerhans' cell
histiocytosis,
haemophagocytic lymphohistiocytosis, pulmonary langerhans cell histiocytosis,
obesity, type
II diabetes, gout, pseudogout, organ transplant rejection, epidermal
hyperplasia, chronic
fatigue syndrome, graft versus host disease (GVHD), lymphadenopathy,
rheumatoid arthritis
(RA), osteoarthritis (OA), inflammatory osteoarthritis, lupus, multiple
sclerosis (MS),
myocarditis, uveitis, CNS disease(s), inflammation aspect to a CNS disease(s),
hypothalamic
inflammation, dementia, glaucoma, amyloid related/driven disease, lipid
storage disease(s),
fibrosis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic
steatohepatitis (NASH),
cirrhosis, cirrhosis of the liver, alcoholic cirrhosis, nephropath(y/ies),
lupus nephritis,
immune nephritis, fibrotic disorder(s), cardiovascular disease, heart disease,
atherosclerosis,
vulnerable plaque, plaque formation, lipid containing macrophage related
disease(s)/disorder(s)/malad(y/ies), macrophage foam cells, diabetes, type 1
diabetes, type 2
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diabetes, insulin resistance, macrophage aspect to insulin resistance,
obesity, obesity
associated inflammation, macrophage accumulation/large numbers of macrophages
in
adipose tissue (e.g. associated with obesity), granuloma(s), granulomatous
diseases,
sarcoidosis (including, without limitation, Annular sarcoidosis, Erythrodermic
sarcoidosis,
Ichthyosiform sarcoidosis, Hypopigmented sarcoidosis, Lofgren syndrome, Lupus
pernio,
Morpheaform sarcoidosis, Mucosal sarcoidosis, Neurosarcoidosis, Papular
sarcoid, Scar
sarcoid, Subcutaneous sarcoidosis, Systemic sarcoidosis, Ulcerative
sarcoidos),
neurosarcoidosis, pulmonary sarcoidosis, interstitial lung disease, pulmonary
fibrosis,
pulmonary tuberculosis, immune reconstitution syndrome of HIV,
Jarisch¨Herxheimer
reaction, sepsis, Paget's disease of bone, osteolysis, monocytosis,
histiocytosis, X-type
histiocytoses, non-X histiocytoses, Langerhans cell histiocytosis, non-
Langerhans-cell
histiocytosis, malignant histiocytosis, malignant histiocytic disorders,
histiocytomas,
histiocytic lymphoma, hemophagocytic syndrome, hemophagocytic
lymphohistiocytosis,
lymphohistiocytosis, diffuse histiocytic sarcoma, Rosai¨Dorfman disease,
gliosis, Bergmann
gliosis, Chronic Obstructive Pulmonary Disease (COPD), chronic inflammatory
lung disease.
In clinical utility, the anti-inflammatory activity of compounds of this
invention juxtaposes
well with their aforementioned ability to reduce body temperature.
.. The anti-inflammatory action by a compound(s) of this invention has an anti-
cancer action.
Because it reduces the number of Tumour Associated Macrophages (TAMs) [155].
These can
constitute a large component of tumour mass and their presence is often
associated with poor
patient prognosis because they can drive cancer pathology. Indeed,
inflammation is now
considered one of the hallmarks of cancer [156]. The anti-inflammatory action,
and thence
.. anti-cancer action, of these compounds synergises with their direct anti-
cancer activities
disclosed herein.
Macrophages can be subverted by pathogens, which hide inside them in safety
from the
immune system. Examples, without limitation, of such pathogens are HIV (causes
HIV/AIDS; HIV virus can lay latent in macrophages during antiretroviral
therapy, wherein
HIV virus becomes undetectable in blood, and then repopulate the virus in
blood when
antiretroviral therapy is interrupted or discontinued; HIV can replicate in
macrophages [157-
1581), Simian Immunodeficiency Virus (Sly), Mycobacterium tuberculosis (causes
tuberculosis), Leishmania parasite (causes Leishmaniasis), Chikungunya virus
(causes
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Chikungunya), Legionella pneumophila (causes Legionnaires' disease),
adenoviruses (e.g. the
adenovirus that causes pink eye), T. whipplei (causes Whipple's Disease),
Brucella spp.
(causes brucellosis), Staphylococcus aureus, Ebola virus, Hepatitis B virus
(HBV), Hepatitis
C virus (HCV), influenza virus strains, dengue virus and antibiotic resistant
bacteria. So, by
exerting anti-macrophage activity, a compound(s) of this invention, or a
pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof, can
treat/ameliorate/prevent/combat such
disorders and diseases. This is especially useful for AIDS, wherein HIV
resides in
macrophages, propagating the immunocompromised character of the disease,
making it more
likely for intracellular opportunistic pathogens (e.g. Mycobacterium
tuberculosis) to reside in
macrophages also. Because the compounds of this invention are selective for
activated
macrophages, an option is to activate macrophages before the compound(s)
administration,
by administering to the patient an effective amount of a compound(s),
protein(s),
antibody(y/ies) or some other entit(y/ies), e.g. pathogen, attenuated pathogen
or pathogen
component that activates macrophages. Some examples (non-limiting) of factors
that can
activate macrophages are cytokines such as interferon-gamma (IFN-y) and/or
tumour necrosis
factor (TNF, TNF-a), and/or IL-4, and/or IL-13, and/or IL-10, and/or IL-2,
and/or IL-12,
and/or IL-6, and/or IL-18 and/or chemokines (CCL3, CCL4, CCL5) and/or a
bacterial
endotoxin such as lipopolysaccharide (LPS), or a commercially available agent
for
macrophage activation in biological research (e.g. CAS 61512-20-7) or an
antibody targeting
a receptor on the macrophage cell surface or on the surface of a different
cell type, which
then activates a macrophage by mechanism. Macrophage activating antibodies are
well
known to those of the art.
An embodiment of this invention is the use of an effective amount of at least
one compound
of this invention, which inhibits FIN ATP hydrolysis, or a pharmaceutically-
acceptable salt,
solvate, hydrate or prodrug thereof, to treat/ameliorate/prevent/combat HIV
infection/transmission/drug resistance, optionally with an effective amount of
a compound(s),
protein(s), antibody(s), pathogen(s) or pathogen component(s) that activates
macrophages
(isn't absolutely necessary because HIV activates macrophages [159-160], which
drives the
chronic inflammation pathology component to HIV infection; HIV stimulates
nitric oxide
production by human macrophages [161, 162, 163, 164]), optionally with a
compound(s),
protein(s), antibody(s), pathogen(s) or pathogen component(s) that polarises
macrophages
towards Ml type (examples, without restriction, include Granulocyte-macrophage
colony-
stimulating factor [GM-CSF] and/or a pharmaceutical
version/analog(s)/biosimilar of GM-
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CSF such as, without limitation, one or more of sargramostim, molgramostim,
regramostim,
filgrastim, pegfilgrastim), optionally in co-therapy with, or after, Anti-
Retroviral Therapy
(ART), combination Anti-Retroviral Therapy (cART, Highly Active Anti
Retroviral
Therapy, HAART), optionally in co-therapy with L-arginine (substrate for
inducible Nitric
Oxide Synthase, iNOS, optionally wherein food(s) rich in this amino acid are
eaten),
optionally for pre- and/or post-exposure prophylaxis (PEP) e.g. after
needlestick injury and/or
sex with an HIV infected person(s), e.g. to reduce the probability of mother
to baby HIV
transmission during pregnancy/birth/breast feeding e.g. given to a subject
without HIV to
decrease the risk they will acquire HIV, optionally wherein this subject is at
sizeable risk of
acquiring HIV, wherein this pre-exposure prophylaxis can reduce the spread of
HIV within a
population (for example in sub-Saharan Africa). Even after prolonged cART,
which drives
plasma HIV down to undetectable levels, HIV-1 DNA and RNA is detectable in
macrophages: they are an HIV reservoir that remains extant, even during cART,
and that the
virus can spread from during any interruption or termination of cART [158].
Moreover, HIV
virus recombines and mutates in macrophages [165], which is a drive to HIV
drug resistance.
Thence the vital importance of the methods and compounds herein. By denying
HIV a
reservoir in which to hide (and mutate, developing drug resistance) from
ART/cART/HAART therapy, a compound(s) of this invention, or a pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof, decreases the amount of
HIV virus in the
body, increasing the chance of HIV viral elimination from the subject,
decreasing the risk the
subject can transmit the HIV virus to another subject, decreasing HIV
associated
symptoms/pathology, decreasing the chance of HIV developing drug resistance to
one or
more drugs used in ART/cART/HAART therapy, improving clinical outcome.
Notably, a
compound(s) of this invention, or a pharmaceutically-acceptable salt, solvate,
hydrate or
prodrug thereof, treats/ameliorates/prevents/combats HIV-associated chronic
inflammation
and/or HIV peripheral neuropathy, wherein the latter is caused by infiltration
of HIV infected
monocytes/macrophages to the dorsal root ganglia (DRG) causing neuronal loss
and
formation of Nageotte nodules.
Macrophages mediate HIV virus neuroinvasion (and neuroinvasion by other
viruses/pathogens also e.g. SARS coronavirus, Heptatitis C virus) and HIV
containing
macrophages and/or microglia cause nervous system and brain patholog(y/ies)
and a
compound(s) of this invention, or a pharmaceutically-acceptable salt, solvate,
hydrate or
prodrug thereof, opposes this and treats/ameliorates/prevents/combats HIV-
associated
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CA 3050553 2019-07-25
neurocognitive disorders (HAND) such as HIV-associated dementia complex (HAD)
(and
neurocognitive and neurodegenerative diseases/disorders caused by other
viruses/bacteria/fungi/parasites/protozoa/pathgens/prions also e.g. [non-
limiting] SARS
coronavirus, cytomegalovirus, herpes simplex encephalitis caused by Herpes
Simplex Virus
(HSV) in microglia, toxoplasmic encephalitis, Japanese encephalitis,
California encephalitis,
tick-borne encephalitis, viral encephalitis {non-limiting e.g. caused by one
or more of rabies
virus, poliovirus, measles virus, Herpesvirus 6, varicella zoster virus,
Epstein-Barr,
cytomegalovirus, coxsackievirus, West Nile virus}, viral or bacterial
meningitis,
meningoencephalitis, encephalomyelitis, refer [166] for further example CNS
pathologies,
without limitation, treated/ameliorated/prevented/combated by a compound(s),
or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, of this
invention). The
anti-HIV and anti-cancer activity of a compound(s) of this invention synergise
to
treat/ameliorate/prevent/combat HIV associated cancers: AIDS-defining cancers
(Kaposi
sarcoma, aggressive B-cell non-Hodgkin lymphoma, cervical cancer) and/or non-
AIDS
defining cancers. This disclosure encompasses a compound(s) of this invention,
or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, in co-
therapy/synergy
with any Food and Drug Administration (FDA) and/or European Medicines Agency
(EMA)
approved drug(s) or treatment(s) for HIV/AIDS or other/additional viral
infection(s).
Examples include, but aren't limited to, abacavir, didanosine, emtricitabine,
lamivudine,
stavudine, tenofovir, disoproxil fumarate (tenofovir DF, TDF), zidovudine
(azidothymidine,
AZT, ZDV), atazanavir, darunavir, fosamprenavir, indinavir, nelfinavir,
ritonavir, saquinavir,
tipranavir, enfuvirtide, maraviroc, dolutegravir, elvitegravir, raltegravir,
cobicistat, acyclovir,
vidarabine. Optionally, additionally, in co-therapy/synergy with any Food and
Drug
Administration (FDA) and/or European Medicines Agency (EMA) approved drug(s)
or
treatment(s) for cancer.
In some invention embodiments a compound(s) of this invention, or a
pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof, is used to
treat/ameliorate/prevent/combat
antibiotic resistant bacteria, optionally intracellular bacteria, optionally
bacteria harbouring
inside macrophages and/or microglia, optionally wherein this intracellular
bacterial reservoir
is contributory to antibiotic resistance. This disclosure encompasses a
compound(s) of this
invention in co-therapy/synergy with any Food and Drug Administration (FDA)
and/or
European Medicines Agency (EMA) approved antibiotic(s)/antimicrobial(s).
Examples
include, but aren't limited to, the following antibiotic classes with an
illustrative non-limiting
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example(s) of each class given in subsequent brackets: Aminoglycosides
(Gentamicin),
Ansamycins (Rifampin), Carbapenems and other penems (Meropenem),
Cephalosporins
(Ceftriaxone), Phosphonic acid derivatives (Fosfomycin), Glycopeptides
(Vancomycin),
Glycylcyclines (Tigecycline), Lipopeptides (Daptomycin), Macrolides and
ketolides
(Erythromycin, telithromycin), Monobactams (Aztreonam), Oxazolidinones
(Linezolid),
Penicillins (Ampicillin, Oxacillin, Amoxicillin), Polymyxins (Colistin),
Quinolones and
Fluoroquinolones (Ciprofloxacin), drugs used solely to treat tuberculosis or
other
mycobacterial diseases (Isoniazid), Amidinopenicillins (Mecillinam),
Amphenicols
(Chloramphenicol), Cephalosporins and cephamycins (Cefazolin), Lincosamides
(Clindamycin), Pleuromutilins (Retapamulin), Pseudomonic acids (Mupirocin),
Riminofenazines (Clofazimine), Steroid antibacterials (Fusidic acid),
Streptogramins
(Quinupristin, dalfopristin), Sulfonamides, dihydrofolate reductase
inhibitors, and
combinations (Sulfamethoxazole, trimethoprim), Tetracyclines
(Chlortetracycline),
Aminocyclitols (Spectinomycin), cyclic polypeptides (Bacitracin),
Nitroftirantoins
(Nitrofurantoin), Nitroimidazoles (Metronidazole).
Macrophages can mediate mother-to-[fetus/baby] viral/pathogen transmission
(vertically
transmitted infection), for (non-limiting) example, zika virus transfers from
mother to fetus
by infecting placental macrophages (Hofbauer cells), HIV-1 virus transfers
from mother to
baby in macrophages in breast milk, and a compound(s) of this invention, or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, opposes
this and
treats/ameliorates/prevents/combats this mother-to-[fetus/baby] viral/pathogen
transmission.
Pathogens can drive macrophages/microglia to drive pathology e.g.
Streptococcus
pneumonia, the most common cause of bacterial meningitis, can cause an
inflammatory
response, triggered by microglia, which can cause intracerebral edema, e.g.
Plasmodium
falciparum, a parasite that causes malaria in humans, can cause cerebral
malaria wherein
microglia have been implicated in this serious malarial complication. A
compound(s) of this
invention or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug
thereof will
treat/ameliorate/prevent/combat a pathogen(s) that drives
macrophages/microglia and/or
another cell of the mononuclear phagocyte system to drive pathology.
Monocytes can be subverted by pathogens, which hide inside them in safety from
the
immune system, wherein this subversion can, in some cases, involve increased
iNOS
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CA 3050553 2019-07-25
expression and increased Nitric Oxide (NO) production in the monocyte.
Examples, without
limitation, of such pathogens include Human Cytomegalovirus (HCMV). So, by
exerting
anti-monocyte activity, especially to those with high intracellular [NO], a
compound(s) of
this invention, or a pharmaceutically-acceptable salt, solvate, hydrate or
prodrug thereof, can
treat/ameliorate/prevent/combat such pathogens and their associated disorders
and diseases.
Non-limiting examples of autoinflammatory diseases/disorders/syndromes that
the
compounds of this invention treat/ameliorate/prevent/combat include, but
aren't limited to,
recurrent fever syndromes, which can be hereditary or acquired, characterized
by recurrent
fever associated with rash, serositis, lymphadenopathy and musculoskeletal
involvement.
Examples include familial mediterranean fever (FMF), TNF receptor-associated
periodic
syndrome (TRAPS), Hyperimmunoglobulinemia D with recurrent fever syndrome
(HIDS),
cryopyrin associated periodic syndrome (CAPS), Blau syndrome, Majeed syndrome,
deficiency of interleukin-1 receptor antagonist (DIRA), mevalonate kinase
deficiency,
pyogenic-arthritis-pyoderma gangrenosum and acne syndrome (PAPA), periodic
fever
aphthous stomatitis pharyngitis adenitis (PFAPA) syndrome, Behcet's disease,
Still's disease,
Crohn's disease, Schnitzler's syndrome, Sweet's syndrome, NLRP12-associated
autoinflammatory disorders, deficiency of interleukin-1 receptor antagonist
(DIRA),
pyoderma gangrenosum, cystic acne, aseptic arthritis, periodic Fever
Associated with
mevalonate kinase deficiency (hyperimmunoglobulin D Syndrome), Pyogenic
Arthritis
Pyoderma Gangrenosum Acne (PAPA) syndrome, Periodic Fever Aphthous Stomatitis,
Pharyngitis and Adenopathy (PFAPA) syndrome, Adult-Onset Still's Disease
(AOSD),
Systemic Juvenile Idiopathic Arthritis (sJIA), Chronic Recurrent Multifocal
Osteomyelitis
(CRMO), Synovitis Acne Pustulosis Hyperostosis Osteitis (SAPHO) syndrome,
Cryopyrin
associated Periodic Syndrome (CAPS), Familial cold auto inflammatory syndrome
(FCAS),
Muckle-Wells syndrome (MWS), Familial cold urticarial, Neonatal onset
multisystemic
inflammatory disorder (NOMID), hereditary Periodic Fever Syndromes, Periodic
Fever
Syndromes, systemic autoinflammatory diseases.
Non-limiting examples of autoimmune diseases/disorders/syndromes that the
compounds of
this invention treat/ameliorate/prevent/combat include, but aren't limited to,
Addison's
disease, Agammaglobulinemia, Alopecia areata, Amyloidosis, Ankylosing
spondylitis, Anti-
GBM/Anti-TBM nephritis, Antiphospholipid syndrome, autoimmune angioedema,
autoimmune dysautonomia, autoimmune encephalitis, autoimmune
encephalomyelitis,
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autoimmune hepatitis, autoimmune inner ear disease (AIED), autoimmune
myocarditis,
autoimmune pancreatitis, autoimmune retinopathy, autoimmune urticaria, axonal
& neuronal
neuropathy (AMAN), BalO disease, Behcet's disease, benign mucosal pemphigoid,
Bullous
pemphigoid, Castleman disease (CD), Celiac disease, Chagas disease, chronic
inflammatory
demyelinating polyneuropathy (CIDP), chronic recurrent multifocal
osteomyelitis (CRMO),
Churg-Strauss, Cicatricial pemphigoid, Cogan's syndrome, cold agglutinin
disease,
congenital heart block, coxsackie myocarditis, CREST syndrome, Berger's
disease,
dermatitis herpetiformis, dermatomyositis, Devic's disease (neuromyelitis
optica), discoid
lupus, Dressler's syndrome, endometriosis, eosinophilic esophagitis (EoE),
eosinophilic
fasciitis, erythema nodosum, essential mixed cryoglobulinemia, Evans syndrome,
fibromyalgia, fibrosing alveolitis, giant cell arteritis (temporal arteritis),
giant cell
myocarditis, glomerulonephritis, proliferative glomerulonephritis, membranous
nephropathy,
minimal change nephrotic syndrome, Goodpasture's syndrome, granulomatosis with
polyangiitis, Graves' disease, Guillain-Barre syndrome, Hashimoto's
thyroiditis, hemolytic
anemia, immune hemolytic anemia, Henoch-Schonlein purpura (HSP), Herpes
gestationis or
pemphigoid gestationis (PG), hypogammalglobulinemia, IgA Nephropathy, IgG4-
related
sclerosing disease, Immune thrombocytopenic purpura (ITP), Inclusion body
myositis (IBM),
Interstitial cystitis (IC), juvenile arthritis, juvenile diabetes (Type I
diabetes), juvenile
myositis (JM), Kawasaki disease, Lambert-Eaton syndrome, Leukocytoclastic
vasculitis,
Lichen planus, Lichen sclerosus, Ligneous conjunctivitis, Linear IgA disease
(LAD), Lupus,
Lyme disease chronic, Meniere's disease, Microscopic polyangiitis (MPA), Mixed
connective tissue disease (MCTD), Mooren's ulcer, Mucha-Habermann disease,
Multiple
sclerosis, Myasthenia gravis, Myositis, Narcolepsy, Neuromyelitis optica,
Neutropenia,
Ocular cicatricial pemphigoid, Optic neuritis, Palindromic rheumatism (PR)
PANDAS,
.. Paraneoplastic cerebellar degeneration (PCD), Paroxysmal nocturnal
hemoglobinuria (PNH),
Parry Romberg syndrome, Pars planitis (peripheral uveitis), Parsonnage-Turner
syndrome,
Pemphigus, peripheral neuropathy, perivenous encephalomyelitis, Pernicious
anemia (PA),
POEMS syndrome, Polyarteritis nodosa, Polyglandular syndromes type I, II, III,
Polymyalgia
rheumatica, Polymyositis, Postmyocardial infarction syndrome,
Postpericardiotomy
syndrome, Primary biliary cirrhosis, Primary sclerosing cholangitis,
Progesterone dermatitis,
Psoriasis, Psoriatic arthritis, Pure red cell aplasia (PRCA), Pyoderma
gangrenosum,
Raynaud's phenomenon, Reactive Arthritis, Reflex sympathetic dystrophy,
Relapsing
polychondritis, Restless legs syndrome (RLS), Retroperitoneal fibrosis,
Rheumatic fever,
Rheumatoid arthritis, Sarcoidosis, Schmidt syndrome, Scleritis, Scleroderma,
Sjogren's
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syndrome, Sperm & testicular autoimmunity, Stiff person syndrome (SPS),
Subacute
bacterial endocarditis (SBE), Susac's syndrome, Sympathetic ophthalmia (SO),
Takayasu's
arteritis, Temporal arteritis/Giant cell arteritis, Thrombocytopenic purpura
(TIP), Tolosa-
Hunt syndrome (THS), Transverse myelitis, Type 1 diabetes, Ulcerative colitis
(UC),
Undifferentiated connective tissue disease (UCTD), Uveitis, Vasculitis,
Vitiligo, Wegener's
granulomatosis (or Granulomatosis with Polyangiitis (GPA)), idiopathic
thrombocytopenia
purpura, splenomegaly.
EXPERIMENTALLY DEMONSTRATING COMPOUND UTILITY AGAINST
ACTIVATED (more than non-activated, resting) MACROPHAGES
This protocol is adapted from that used in [154]. It is illustrative and not
limiting. Alternative
protocols, and modification(s) to this protocol, will be clear to those of the
art, based on the
common general knowledge, the literature, and/or the content of prior art
disclosures cited
herein. Cells of the murine macrophage cell line J774.A1(ATCC TIB67) are
adherent. Very.
Physical (e.g. scraping/lifting) and/or chemical techniques (e.g. one or more
of EDTA, EDTA
in PBS solution, trypsin, TrypLE, HyQTase, Detachin, Accutase, Versene
solution,
nonenzymatic cell dissociation solution from Sigma-Aldrich, Cellstripper
nonenzymatic cell
dissociation solution or {Dulbecco's phosphate-buffered saline [DPBS] with no
calcium and
no magnesium solution}) can separate them from an adhered surface but this
manipulation(s)
risks damaging/modifying the macrophages. Adherence is lessened by culturing
macrophages
in a container with a Teflon coated surface (e.g. Teflon coated
(polytetrafluoroethylene)
flasks or gas permeable bags that are coated with Teflon or Teflon like
material made by
companies such as Milteny, OriGen, Gibco, CellGenix [FEP Teflon-coated cell
culture bag],
Afc and others) or (less preferably) in bacterial plastic petri dishes (i.e.
non cell culture
coated) or polypropylene plasticware. In the preferred protocol, adherence
issues are
circumnavigated because J774.A1 macrophages are grown in suspension in stirrer
bottles
(Techne, Burlington, NJ, USA; stirring lifts the cells into suspension, low
speed [12-25 rpm]
to minimize cell shearing, soft-start/stop feature provides slow acceleration
and deceleration
of the stirrer, low heat production, its glass is siliconized to reduce cells
adhering to it)
containing cell culture medium (Dulbecco's modified Eagle's medium, DMEM)
supplemented with 10% Fetal Calf Serum (FCS), 4.5 g/1D-glucose, 2 mM L-
glutamine (4
mM final), 25 mM HEPES, penicillin (100 units/ml) and streptomycin (100 g/ml)
{optionally L-arginine is also added, e.g. at 400 M in final medium, wherein
this is a
substrate that iNOS uses to produce nitric oxide [NO]}. Incubated at 37 C in a
humidified
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atmosphere containing 5% CO2. Cell density is kept <1*106 cells/ml, which
maintains cell
viability at >98%. Four samples are taken from this suspension culture and
each re-suspended
in a separate stirrer bottle in fresh medium at a density of 0.2*106 = 200,000
cells/ml. Two of
these samples additionally contain 10 U/ml murine IFN-y (e.g. sourced from
Insight Biotech,
.. Wembley, UK) and 10 ng/ml LPS (LPS from Salmonella typhosa 0901 e.g.
sourced from
Difco, Surrey, UK), which activates J774.A1 macrophages. So, there are two
samples in
which the macrophages are not activated, and two samples in which the
macrophages are
activated. Test drug is added to a non-activated and an activated sample,
wherein drug
concentration in medium is 10 iM (optionally different, to illustrate, 1 or
100 M), drug
vehicle is added to the two other samples. The test drug is a compound of this
invention, an
FIR ATP hydrolysis inhibitor, which preferentially inhibits FIFO ATP
hydrolysis over FiFo
ATP synthesis, for example a compound of Formula (I), (II), (III), (IV), (V)
or (VII) or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
optionally Compound
7b of this disclosure. Vehicle is DMSO or [12.5% solutol, 12.5% ethanol, 75%
water], or
[25% solutol and 75% water], or some other vehicle selected by one of the art.
So there are
four different samples, numbered as: (1) non-activated+vehicle, (2) non-
activated+drug, (3)
activated+vehicle, (4) activated+drug. At 3 hour time points, for 12 to 24
hours, aliquots (e.g.
of 15 ml each) are taken from each of the four samples and cell viability in
each aliquot is
recorded using the Trypan blue exclusion method. A further embodiment is to
record for a
longer period, wherein if longer than 24 hours, optionally the medium is half-
changed for all
samples (optionally wherein drug/vehicle is at same concentration in replacing
medium than
medium replaced) every 24 hours to avoid nutrient depletion and medium
acidification,
which can happen with activated macrophages especially because they have a
high glycolytic
metabolism. Optionally the study is run with replicates of each sample and/or
the study is run
more than once and/or the study is run again with different drug
concentration(s) and/or the
study is run with RAW 264.7 mouse macrophages (ATCC TIB-71; also an adherent
cell line)
instead of J774.Al.
During the course of the study, the macrophage number in Sample 1 increases.
The gradient
of increase is less, or negative, for Sample 2 because compounds of this
invention are anti-
cancer therapeutics and J774.A1 is a cancer cell line. Upon activation,
J774.A1 macrophages
stop proliferating and so the macrophage number of Sample 3 is constant or
slightly
decreases during the study. Upon activation, J774.A1 macrophages produce
substantial
amounts of nitric oxide (NO), which reduces their OXPHOS use, and so they
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disproportionally rely upon F1F0 ATP hydrolysis to maintain Tim, wherein a
compound of
this invention selectively inhibits FIR, ATP hydrolysis, Tim collapses,
apoptosis follows.
Sample 4 will have a much lower number of macrophages than the other samples.
Although
the Sample 4 number is greater if S-ethyl isothiourea (SEITU, 500 NI in
medium, available
from Sigma-Aldrich) is administered coincident with macrophage activation,
because SEITU
inhibits inducible NO synthase (iNOS). Sample 4 having a lower macrophage
number than
Sample 3 shows that a compound of this invention, optionally 7b, can reduce
the number of
activated macrophages. Sample 4 having a lower macrophage number than Sample 2
shows
that a compound of this invention, optionally 7b, disproportionally affects
activated rather
than resting macrophages. The amplitude of this differential, between Sample 2
and Sample
4, isn't as large as it could be because J774.AI is a cancer cell line and a
compound of this
invention, e.g. 7b, also exerts anti-cancer activity. A bigger differential
between Sample 2
and Sample 4 can be observed if primary macrophages (M1 type, iNOS expressing,
nitric
oxide (NO) producing) are used in this study instead of a cancer cell line.
The following
protocol is adapted from that used in [167], with background detail added from
[168].
Peritoneal macrophages are isolated from 8 week old pathogen free CD-1 mice.
An eliciting
agent, such as 1.5 ml of sterile thioglycolate broth (Difco Laboratories)
intraperitoneal
injected 4 days before macrophage sourcing, which increases monocyte migration
into the
peritoneum, therefore increasing macrophage yield, is NOT used, because this
can activate
the macrophages, causing them to produce nitric oxide [167], whereas
resting/unactivated
macrophages are desired. So, resident rather than elicited macrophages are
sourced from mice
peritoneum, wherein the yield is around ¨0.5-1*106 macrophages per mouse (-1-
2*106 total
peritoneal exudate cells, ¨40% are macrophages). With these sourced cells, the
macrophages
are separated from other cell types by flow cytometry coupled with cell
sorting or purification
.. using magnetic beads and then cultured at a seeding density of 300,000
cells/ml, at 37 C in a
humidified atmosphere containing 5% CO2, cultured in DMEM (Sigma¨Aldrich)
supplemented with 10% (v/v) fetal bovine serum, 100 U/ml penicillin, 100 g/ml
streptomycin and 250 ng/ml amphotericin B. Wherein these macrophages can be
activated by
100 ng/ml LPS (from Escherichia coli strain 111:B4) and IFN-y (100 U/ml).
These primary
macrophages can be used in place of J774.A1 cells in the aforementioned
protocol. [168]
describes alternative methods to source primary macrophages from mice, wherein
a preferred
method is to use a method it describes but with GM-CSF instead of the M-CSF
that it
specifies. Granulocyte-macrophage colony-stimulating factor (GM-CSF) promotes
M1
whereas supplementation with Macrophage colony-stimulating factor (M-CSF)
promotes M2
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macrophages [169]. Although activating with LPS and IFN-y favours M1
polarization of
primary macrophages anyhow. It is important that the primary macrophages used
produce
nitric oxide (NO) when activated, wherein NO production can optionally be
checked for, in a
pre-experiment and/or during the test drug application and control
experiments, using a
method of the art e.g. using Griess reagent (1:1 v:v with culture medium,
Sigma-Aldrich) and
the Griess reaction wherein absorbance at 540 or 570 nm is assayed, e.g. using
an ELISA
reader (Bio-Tek Instruments, Burlington, VT; standard curves of various fixed
concentrations
of sodium nitrite (Sigma-Aldrich) are used to calibrate a given absorbance to
a given nitrite
concentration) after 20 minutes of reaction time. Or, alternatively, or in
addition, an NO
nanosensor (amiN0-600, Innovative Instruments, FL) can be used. Or, or in
addition,
Electron Paramagnetic Resonance Detection of Free NO by spin-trap method can
be used
[170]. An easier option is to source cryopreserved primary mouse macrophages
(e.g. from
C57BL/6 mice) for this experiment commercially, for (non-limiting) example
from Astarte
Biologics LLC (Bothell, WA, USA) or Nanobiotech (Whippany, NJ, USA).
A lesser preferred study option is to, instead of culturing the (mouse primary
or mouse cancer
cell line {e.g. J774.A1}) macrophages in suspension using stirrer bottles
(Techne, Burlington,
NJ, USA; can be expensive) as has been described, permit the macrophages to
adhere to a
surface, wherein their cell viability is assayed using the RealTime Glo MT
Cell Viability
.. Assay (Promega, Madison, WI), which can assay the viability of adhered
cells and so doesn't
require prior scraping/lifting etc. that itself can negatively modulate
macrophage cell number,
wherein this assay reports ATP content, wherein macrophage activation itself
decreases
intracellular [ATP] [154], and so this must be factored into interpretation
(ATP content
doesn't just scale with cell viability, but can correlate with cellular
processes also, and so this
.. [ATP] assay isn't a direct read out of cell number, which often isn't
appreciated by those of
the art, however, in this case this feature of the assay doesn't change the
study conclusion:
activation of macrophages decreases their ATP content without test drug
applied, with test
drug of this invention applied, e.g. compound 7b, the ATP content decreases
even more after
activation because the test drug drives activated macrophages to death,
reducing the
macrophage number). In this alternative study plan, commercially sourced
cryopreserved
bone-marrow macrophages from C57BL/6 mice are thawed, counted and plated in a
96 well
flat bottom plate at 2*104 cells/well (well volume is 0.2 ml, thus macrophage
density =
0.1*106 = 100,000 cells/m1) in Iscove's Modified Dulbecco's Medium (IMDM) + 2
mM
Glutamaxlm (a dipeptide: L-alanine-L-glutamine) + 10% Fetal Calf Serum (FCS) +
20 ng/ml
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M-CSF (or more preferably GM-CSF). The plate is transferred to a 37 C, 5% CO2
incubator
for 18-24 hours. In a pre-experiment, optionally using a commercially sourced
nitric oxide
assay kit, it is assessed whether the sourced macrophages produce nitric oxide
when activated
(by LPS {100 ng/m11 and IFN-y {100 U/ml} added) in this experimental set up,
wherein it is
necessary that they do, and if not the source of macrophages is changed and/or
the activation
protocol is changed. Furthermore, it is also assessed whether the sourced
macrophages
produce nitric oxide when NON-activated (resting), wherein it is necessary
that they do not,
at least not to any major degree. There should be an observed NO production
differential
between the resting and activated macrophages, wherein activated should
produce much more
NO than resting macrophages. In the actual experiment, test drug (10 M;
experiment is run
other times with 1 or 100 M, or other concentration(s)) and/or [LPS {100
ng/m11 and IFN-y
{100 U/m1}} is added after the 18-24 incubation period, as shown in the table
below:
LPS IFN-y Test drug
1 Non-activated+vehicle No No No
2 Non-activated+Test drug No No Yes
3 Activated+vehicle Yes Yes No
4 Activated+Test drug Yes Yes Yes
After macrophage activation and/or test drug addition, macrophage cell
viability
(approximated by reading out [ATP]) is recorded by RealTime Glo MT Cell
Viability Assay
(Promega, Madison, WI) at 0, 3, 6, 9 and 24 hours. Optionally there are two
more test groups,
wherein SEITU drug (500 NI in medium in well), which inhibits iNOS, is added:
LPS IFN-y Test drug SEITU
5 Activated+vehicle+SEITU Yes Yes No Yes
6 Activated+vehicle+SEITU+Test drug Yes Yes Yes Yes
.. EXPERIMENTALLY DEMONSTRATING COMPOUND UTILITY AGAINST HIV
INFECTED MONOCYTES/MACROPHAGES
This protocol is adapted from that used in [170]. It is illustrative and not
limiting. Alternative
protocols, and modification(s) to this protocol, will be clear to those of the
art, based on the
common general knowledge, the literature, and/or the content of prior art
disclosures cited
herein. Human monocytes are recovered from peripheral blood mononuclear cells
(PBMCs)
of an HIV- and hepatitis B- seronegative human donor after leukapheresis and
purified by
countercurrent centrifugal elutriation. Cell suspensions are >98% monocytes by
the criteria of
cell morphology on Wright-stained cytosmears, by granular peroxidase, and by
nonspecific
esterase. Monocytes are cultured, at 37 C in a humidified atmosphere
containing 5% CO2, in
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Teflon coated (polytetrafluoroethylene) flasks {or gas permeable bags that are
coated with
Teflon or Teflon like material made by companies: Milteny, OriGen, Gibco,
CellGenix [FEP
Teflon-coated cell culture bag], Afc and others} (at 2*106 cells/ml) in DMEM
(Sigma
Chemical Co., St. Louis, MO) supplemented with 10% pooled human sera, 50 g/ml
gentamicin, and 1,000 U/ml highly purified (<0.01 ng/ml endotoxin) recombinant
human M-
CSF {or alternatively GM-CSF} (Genetics Institute, Cambridge, MA) for 7 days
before HIV
virus infection. HIV Infection of Monocytes: the monocyte culture is exposed
to 2*105
TCID5o/m1 of a monocytotropic viral strain, HIV-1A, wherein all viral stocks
are tested
prior and found free of mycoplasma contamination (Gen-probe II; Gen-probe
Inc., San
Diego, CA). Culture medium is half-exchanged every 2 days. On the 6th day
(optionally
earlier or later, optionally after significant Reverse Transcriptase activity
is recorded in the
medium, assayed as described in [170] and references, particularly its
reference 6), four
samples are taken from this suspension culture and each re-suspended in a
separate Teflon (or
Teflon like) coated flask/bag in fresh medium at a density of 0.2*106
cells/ml. Two of these
samples are administered with LPS (10 ng/ml) and TNF-a (1000 U/ml). Test drug
is added to
a sample with and without [LPS and TNF-a], wherein drug concentration in
medium is 10
M (optionally different, to illustrate, 1 or 100 M), drug vehicle is added to
the two other
samples. The test drug is a compound of this invention, an FIFO ATP hydrolysis
inhibitor,
which preferentially inhibits FIFO ATP hydrolysis over Fi Fo ATP synthesis,
for example a
compound of Formula (I), (II), (III), (IV), (V) or (VII) or a pharmaceutically-
acceptable salt,
solvate, hydrate or prodrug thereof, optionally Compound 7b of this
disclosure. Vehicle is
DMSO or 12.5% solutol, 12.5% ethanol, 75% water, or 25% solutol and 75% water,
or some
other vehicle selected by one of the art. So there are four different samples,
numbered as: (1)
vehicle, (2) drug, (3) [LPS and TNF-a]+vehicle, (4) [LPS and INF-a]+drug. At 3
hour time
points, for 12 to 24 hours, aliquots (e.g. of 15 ml each) are taken from each
of the four
samples and cell viability in each aliquot is recorded using the Trypan blue
exclusion method.
A further embodiment is to record for a longer period, wherein if longer than
24 hours,
optionally the medium is half-changed for all samples (optionally wherein
drug/vehicle is at
same concentration in replacing medium than medium replaced) every 24 hours to
avoid
nutrient depletion and medium acidification. Optionally the study is run with
replicates of
each sample and/or the study is run more than once and/or the study is run
again with
different drug concentration(s). Importantly the study is repeated with a
different monocyte
donor used, wherein the study is repeated 20 times with 20 different human
donors.
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The monocyte number is less in Sample 2 than in Sample 1, showing that a
compound of this
invention kills HIV infected monocytes. HIV infected monocytes, in distinction
to uninfected
monocytes, produce nitric oxide, which reduces their OXPHOS use, and so they
disproportionally rely upon Fi Fo ATP hydrolysis to maintain Tim, wherein a
compound of
this invention selectively inhibits FIFO ATP hydrolysis, Tim collapses,
apoptosis follows. The
monocyte number is even less in Sample 4 than Sample 2, showing that [LPS and
1NF-a]
assist the killing of HIV infected monocytes by a compound of this invention,
which they do
by increasing nitric oxide production by I-IIV infected monocytes further,
which further
reduces their OXPHOS use, further increases their reliance upon Fi Fo ATP
hydrolysis to
.. maintain TN, wherein a compound of this invention selectively inhibits FiFo
ATP hydrolysis,
Tim collapses, apoptosis follows. There may be no difference between Sample 4
and Sample
2 if, in Sample 2, the nitric oxide production is already at maximal capacity.
Optionally, the
nitric oxide dependence can be shown by running the study again but
administering the test
drug along with NG-methyl-L-arginine (2 mM), a competitive inhibitor of
inducible Nitric
.. Oxide Synthase (iNOS, competing with L-arginine), wherein the difference in
monocyte
number between Sample 1 and 2 will be less or absent.
Control: testing drug effect on monocytes that haven't been infected with HIV:
referring to
the aforementioned protocol: just before the HIV infection step, two samples
are taken from
the uninfected suspension culture and each re-suspended in a separate Teflon
(or Teflon like)
coated flask/bag in fresh medium at a density of 0.2*1V cells/ml. Test drug in
vehicle is
added to one sample, wherein drug concentration in medium is 101AM (optionally
different,
to illustrate, 1 or 100 0/1), drug vehicle alone is added to the other sample.
At 3 hour time
points, for 12 to 24 hours, aliquots (e.g. of 15 ml each) are taken from each
of the four
.. samples and cell viability in each aliquot is recorded using the Trypan
blue exclusion method.
A further embodiment is to record for a longer period, wherein if longer than
24 hours,
optionally the medium is half-changed for both samples (optionally wherein
drug/vehicle is at
same concentration in replacing medium than medium replaced) every 24 hours to
avoid
nutrient depletion and medium acidification. In this case, the monocyte number
isn't
.. especially different between the drug and vehicle only treated samples.
None of these
monocytes are activated, and so don't produce nitric oxide substantially, and
so there is no
particular reliance upon Fi Fo ATP hydrolysis to maintain Tim, thence no
particular
sensitivity/liability to a compound of this invention.
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EXPERIMENTALLY DEMONSTRATING COMPOUND UTILITY AGAINST HIV
INFECTED MACROPHAGES
This protocol is adapted from that used in [171]. It is illustrative and not
limiting. Alternative
protocols, and modification(s) to this protocol, will be clear to those of the
art, based on the
common general knowledge, the literature, and/or the content of prior art
disclosures cited
herein. The buffy coat is the fraction of an anticoagulated blood sample that
contains most of
the white blood cells and platelets following density gradient centrifugation
of the blood.
Buffy coats from (at least 10) healthy donors are obtained. Peripheral blood
mononuclear
cells (PBMCs) are isolated from buffy coats by Percoll or Ficoll-Paque Plus
(GE Healthcare
.. Life Sciences, Pittsburgh, PA) gradient centrifugation. Monocytes are
isolated from PMBCs
using CD14 microbeads (Miltenyi Biotec, Auburn, CA). Monocyte purity is
confirmed by
fluorescence-activated cell sorter (FACS) analysis. Then, to generate
macrophages, the
monocytes are cultured for 7 days to allow differentiation into macrophages:
optionally
cultured in 50% Myelocult (Stemcell Technologies, Vancouver, Canada), 25%
Iscove's
modified Dulbecco's medium (IMDM) (containing 10% FBS), 25% H527 human
fibroblast
conditioned IMDM, and 1 ng/ml of macrophage colony-stimulating factor (M-CSF,
or
alternatively, more preferably, GM-CSF) (Sigma-Aldrich, St. Louis, MO). After
7 days
culture, wherein the starting monocytes are now macrophages, the macrophages
are cultured
at 37 C in RPM! 1640 medium supplemented with 10% fetal bovine serum (FBS) and
1mM
sodium pyruvate at 45,000 cells/ml (9,000 cells per well) in triplicate in a
96-well plate in the
presence of various concentrations (e.g. 0.1, 1, 10, 100 tiM) of test drug,
wherein this drug is
a compound of this invention, optionally Compound 7b. After 24 hours, the
CellTiter-Glo
reagent is added, resulting in cell lysis and generation of a luminescent
signal proportional to
the amount of ATP present in each well. The data is normalized to the average
signals from
the untreated (no drug added) cells. This investigation aspect shows that the
drug is non-toxic
to normal non-activated macrophages at a concentration(s) that, in the next
step, is shown to
reduce the number of macrophages infected with an HIV reporter virus. The
absence of drug-
associated cytotoxicity at 24 hours is also confirmed by trypan blue
exclusion.
The HIV-1 reporter virus construct pSF162R3 Nef+ plasmid is replication
competent,
macrophage-tropic and expresses Enhanced Green Fluorescent Protein (EGFP) in
conjunction with HIV nef gene expression and has been used to track HIV
expression in
macrophage cultures in prior studies in the literature. Detection of GFP
protein is indicative
of Tat-mediated transactivation of the HIV promoter and the expression of
early viral gene
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products such as Nef, Tat and Rev. Therefore, GFP detection can serve as a
surrogate marker
for HIV DNA integration and Nef expression. The plasmid is transformed into
Max
Efficiency Stb12 competent cells (Life Technologies, Grand Island, NY); large-
scale plasmids
are prepared using the PureYield plasmid MaxiPrep system (Promega, Madison,
WI). The
viral stocks are generated by transient transfection into 293T/17 cells (ATCC,
Manassas, VA)
using SuperFect transfection reagents (Qiagen, Valencia, CA) according to the
manufacturer's protocol. At 37 C, macrophages are infected with viral
supernatant at 150 to
180 ng p24 per million cells (0.45 to 0.75 pg per well, 6 to 8 M cells per
well) for 6 hours in
the presence or absence of test drug, a compound of this invention.
Subsequently, the viral
inoculate is removed, and the cells are washed and cultured in the same medium
for 7 days
with or without drug treatment. Medium is exchanged every 3 to 4 days. Cells
are inspected
for EGFP expression before harvesting using a fluorescence microscope. When
examined
under a fluorescence microscope, observed is a significant reduction in the
proportion of
EGFP-HIV+ macrophages following treatment with test drug. Macrophages are
harvested by
scraping and then stained using the LIVE/DEAD fixable red dead cell staining
kit (Life
technologies, Grand Island, NY) according to the manufacturer's manual,
followed by
fixation in 3% formaldehyde. The samples are analysed on a FACScan instrument
and the
percentage of EGFP+ cells are determined after gating out the debris by
FSC/SCC and the
dead cells by the LIVE/DEADstain. Observed is a significant reduction in the
proportion of
live EGFP-HIV+ macrophages, and/or less ELISA quantification of supernatant
HIV p24
antigen, when treated with test drug.
SLOW RELEASE FORMULATIONS
An invention embodiment is to administer to a subject a therapeutic amount of
at least one
compound of this invention, for example a compound of Formula (I), (II),
(III), (IV), (V) or
(VII), in a formulation/dosage selected from modified release, extended
release, long acting
release, sustained release, prolonged release, controlled release, slow
release or similar, as
clear to someone of the art, for use in a method of treatment of the human or
animal body by
therapy. Such a formulation exposes the subject body to the compound(s) over a
longer
period of time than if the compound was applied alone. This is useful because
it delivers
good area under the curve for the compound, which for example exerts anti-
cancer activity in
the subject, without an abrupt large body temperature drop. Any body
temperature drop is
less in amplitude, more in duration, which is safer. Methods to make such
formulations for a
compound are well known to those of the art. To illustrate with some non-
limiting examples:
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compound is administered in an excipient/tablet/pill which takes time to
dissolve/degrade/disintegrate because it is, for example, poorly soluble.
Furthermore, enteric
coating, acrylics (e.g. chitin), liposomes, drug-polymer conjugate(s),
microencapsulation
(coating an active pharmaceutical ingredient around an inert core and layering
it with
insoluble substances to form a microsphere), dissolution systems (rate release
is dependent on
dissolution of an excipient; 2 categories: reservoir, matrix), diffusion
systems (rate release is
dependent on rate that drug dissolves through a barrier, usually a type of
polymer; 2
categories: reservoir, matrix), osmotic systems, ion exchange resins, matrix
systems (sub-
categories: hydrophobic matrices, lipid matrices, hydrophilic matrices,
biodegradable
matrices, mineral matrices), stimuli induced release (e.g. temperature,
ultrasonic, electronic
etc.) and other encapsulation technologies known to those of the art.
An invention embodiment is using a dosage regime of FIFO ATP hydrolysis
inhibitor that
doesn't cause a significant temperature drop in a human. Optionally spreading
the daily
therapeutic dose over multiple pills per day so that any body temperature drop
is lowered in
amplitude, lengthened in duration, which is safer.
ALBUMIN AS SLOW RELEASE ASSIST
Compound 31 of this invention is a 1,4-benzodiazepine, which is a compound
class known to
bind albumin in blood extensively. For example, 99% of diazepam is protein
bound in the
blood, wherein the overwhelming majority of this protein is albumin. Albumin
can bind
many things non-specifically but it has high affinity binding sites with
higher affinity binding
to certain substances [172], for example, 1,4-benzodiazepines. Indeed, albumin
has two
specific high affinity binding sites, one of which is called the
"benzodiazepine site", also
called Site II, the diazepam site or the indole-BDZ site, which can bind a
range of
benzodiazepines, and so possibly Compound 31 also. Compound 31 binding albumin
in
blood will buffer, slow and prolong Compound 31 exposure to the tissues, which
will buffer,
reduce the amplitude, slow and prolong Compound 31 effect on body temperature.
In the
anti-cancer data of this disclosure, Compound 31 (Figure 6) outperformed the
FDA
approved, widely used, chemotherapeutic carboplatin (Figure 1). However, it
underperformed by comparison to its EC50 FIFO ATP hydrolysis (Figure 9) and
its
sequestration by binding albumin in the 5% fetal bovine serum (FBS) added to
RPMI 1640
medium used in NCI-60 testing [35] might be contributory to this
underperforrnance (in
addition to Compound 31 interaction with CYP2C9, as cited in Figure 9 and
legend).
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Reducing FBS amount to 2% and assaying if Compound 31 exerts greater anti-
cancer activity
vs. no drug control (also with FBS reduced to 2%) would reveal this. N.B.
carboplatin does
not bind albumin with any great affinity, indeed, its protein (i.e. mainly
albumin) binding in
blood is very low.
An invention embodiment is to administer a compound(s) of this invention, one
or more of a
FiFo ATP hydrolysis modifier, optionally a compound(s) of Formula (I), (II),
(III), (IV), (V),
(VI) and/or (VII), in complex with albumin to a subject, optionally for use in
a method of
treatment of the human or animal body by therapy, optionally to
treat/ameliorate/prevent/combat one or more diseases/disorders/conditions
referred to in this
disclosure, optionally cancer, optionally for the
treatment/amelioration/prevention/combat of
cancer in a subject(s), and/or for the manufacture of a medicament, optionally
for treating one
or more of the diseases/disorders/conditions referred to in this disclosure,
optionally cancer.
TEMPERATURE CONTROLLED RELEASE
An invention embodiment is a temperature-sensitive pharmaceutical
composition/vehicle that
only releases a compound of this invention, for example a compound of Formula
(I), (II),
(III), (IV), (V), (VII) or other FIFO ATP hydrolysis inhibitor(s), when the
body is at normal
body temperature or higher. The latter is reached if the subject has a fever
for example. Many
cancers cause fever. Such a temperature-sensitive delivery
composition/vehicle, releasing
drug(s) at normal body temperature (37 C) for example, can effect a safety
feedback loop
because as FiFo ATP hydrolysis inhibitor is released, body temperature falls,
thence less drug
is released, body temperature can thence recover, further compound is
released, and this loop
iterates, implementing extended release and minimising the perturbation to
body temperature
from optimal. For non-limiting example, a FIFO ATP hydrolysis inhibitor(s) is
loaded into a
structure incorporating biocompatible thermo-sensitive polymer which shrinks
at a
temperature exceeding its phase/volume transition temperature, releasing the
compound. This
volumetric change is reversible. Should the temperature subsequently fall
below the
phase/volume transition temperature the structure expands and compound release
doesn't
occur [173]. In some embodiments the phase/volume transition temperature is
tuned to be at
normal body temperature, in other embodiments, at a pathologically elevated
body
temperature(s). Biocompatible thermosensitive polymers can be used to prepare
temperature-
responsive hydrogels/nanogels and thence nanoparticles, optionally with
polysaccharides to
modulate the drug encapsulation and release efficiency, which have a phase
transition
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temperature, above which they release the "cargo" compound(s). Transition
temperature can
be readily tuned by the copolymerization conditions and by varying the content
of repeating
units in the copolymer. Non-limiting options for making temperature sensitive
vehicles for
compounds of this invention include thermosensitive hydrogels/nanogels,
temperature
sensitive liposomes [174-176] (these have been used in clinical trials e.g.
ThermoDox),
thermosensitive micelles, polymeric micelles, core shell structures,
core¨shell microgel
particles, thermoresponsive composite films, smart three dimensionally ordered
porous
materials, thermosensitive microcontainers, nanoscale drug delivery vehicles.
Also contemplated and componentry to this invention is a compound(s) of this
invention
administered/released by a pharmaceutical composition/vehicle triggered by
temperature in
parallel with one or more other stimuli e.g. pH, and/or a pharmaceutical
composition/vehicle
that disproportionally delivers a compound(s) of this invention to cancer as
compared to
normal tissue(s) in a subject (numerous strategies available to those of the
art e.g. refer
[177]). For example, by the pharmaceutical composition/vehicle release being
triggered by
one or more cancer associated stimuli e.g. acidic pH, or one or more
externally applied
stimuli to the cancer/tumour e.g. heat.
COMPOUND 31, CYP2C9
Compound 31 inhibits CYP2C9 [8], which decreases Compound 31 inhibition of
FIFO ATP
hydrolysis, and an invention embodiment is a method of treating, ameliorating,
preventing or
combating a disease or disorder, optionally cancer, by administering to a
subject a
therapeutically effective amount of Compound 31, and/or another F IF ATP
hydrolysis
inhibitor, optionally a compound(s) of Formula (I), (II), (III), (IV), (V),
(VI), (VII) and a
CYP2C9 substrate(s) and/or inhibitor(s), optionally wherein these compounds
are in the same
pharmaceutical composition or sold/distributed together, optionally in the
same packaging.
An invention embodiment is Compound 31, and/or another FIFO ATP hydrolysis
inhibitor,
optionally a compound(s) of Formula (I), (II), (III), (IV), (V), (VI), (VII)
and a CYP2C9
substrate(s)/inhibitor(s), and/or other cytochrome P450 enzyme
substrate(s)/inhibitor(s), for
use in a method of treatment of the human or animal body by therapy, wherein
the latter is,
illustratively, not restrictively, one or more of fluorouracil (5-FU),
capecitabine (prodrug of
5-FU), imatinib (gleevac), nilotinib, sorafenib, dasatinib, teniposide,
tamoxifen, idarubicin,
sulfaphenazole, fluconazole, miconazole, cannabis, THC, a coumarin or coumarin
derivative/analogue, amentoflavone, valproic acid, valproate, apigenin,
amiodarone,
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antihistamines, cyclizine, promethazine, chloramphenicol, fenofibrate,
flavone, flavonol,
fluvastatin, fluvoxamine, isoniazid, lovastatin, modafinil, phenylbutazone,
probenecid,
sertraline, sulfamethoxazole, voriconazole, zafirlukast, quercetin,
tetrahydrocannabinol,
cannabidiol, cannabinol, polyunsaturated fatty acids, nonsteroidal anti-
inflammatory drug
.. (NSAID), celecoxib, lornoxicam, diclofenac, ibuprofen, flubiprofen,
naproxen, ketoprofen,
piroxicam, meloxicam, suprofen, phenytoin, fluvastatin, sulfonylureas,
glipizide,
glibenclamide, glimepiride, tolbutamide, glyburide, angiotensin II receptor
antagonists,
irbesartan, losartan, S-warfarin, sildenafil, terbinafine, amitriptyline,
fluoxetine, nateglinide,
rosiglitazone, tamoxifen, torasemide, JWH-018, AM-2201, limonene, tapentadol,
montelukast, ticrynafen, benzydamine N-oxide hydrogen maleate, phenprocoumon,
benzbromarone, acenocoumarol, methadone, clopidogrel, delavirdine, disulfiram,
doxifluridine, efavirenz (HIV/AIDS drug), leflunomide, metronidazole,
sulfinpyrazole,
nicardipine, gemfibrozil, floxuridine, levofloxacin, progesterone,
testosterone etc.
.. A preferred method to treat/ameliorate/prevent/combat cancer in a subject
is to administer
Compound 31, and/or another FiFo ATP hydrolysis inhibitor, optionally a
compound(s) of
Formula (I), (II), (Ill), (IV), (V), (VI), (VII) for example Compound 24 from
[8] (ICso
CYP2C9 = 0.038 M, IC50 FiFo ATP hydrolysis = 0.008 M), and a
chemotherapeutic(s) that
is a substrate/inhibitor of CYP2C9 (non-limiting e.g. one or more of
fluorouracil (5-FU),
capecitabine (prodrug of 5-FU), imatinib (gleevac), nilotinib, sorafenib,
dasatinib, teniposide,
idarubicin, tamoxifen etc.); these drugs interfere with the metabolism of each
other and cause
greater anti-cancer activity per unit dose of each administered drug than when
each drug is
administered alone. Furthermore, when an anti-pain or anti-inflammatory
drug(s) is
prescribed, an invention embodiment is that it should be a drug that is CYP2C9
metabolised
e.g. celecoxib, tapentadol, ibuprofen etc. An invention embodiment is a method
wherein the
subject's CYP2C9 gene sequence is obtained/read/analysed, and/or the sequence
of their
CYP2C9 regulatory non-coding DNA regions, and/or their CYP2C9 gene/protein
expression
is measured, in order to inform at what dosage, if at all, Compound 31, and/or
another FiFo
ATP hydrolysis inhibitor, optionally a compound(s) of Formula (I), (II),
(III), (IV), (V), (VI),
(VII) should be administered to a subject for use in a method of treatment of
the human or
animal body by therapy. Impaired CYP2C9 function in a subject, either because
of gene
sequence and/or expression (non-limiting e.g. human subject with rs1799853(T)
and/or
RS1057910(C) and/or RS9332131 (a frame shift) SNP variants of CYP2C9), permits
lower
Compound 31 dosage, and/or lower dosage of another FIR' ATP hydrolysis
inhibitor,
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optionally a compound(s) of Formula (I), (II), (III), (IV), (V), (VI), (VII)
to be used in a
method of treatment of the subject by therapy.
An invention embodiment is to administer Compound 31 to a subject, and/or
another FiFo
ATP hydrolysis inhibitor, optionally a compound(s) of Formula (I), (II),
(III), (IV), (V), (VI)
and/or (VII), with an anti-coagulant(s) (e.g. optionally to
treat/ameliorate/prevent/combat one
or more of deep vein thrombosis, pulmonary embolism, systemic embolism, venous
thromboembolism, acute coronary syndrome, haemorrhage, stroke, ischemic
attack,
thrombotic disorder, coronary artery disease, myocardial infarction, atrial
fibrillation,
thrombophilia etc.) that is not warfarin, especially not S-warfarin, wherein
administered anti-
coagulant(s) embodiments include one or more of heparin, unfractionated
heparin, low-
molecular-weight heparin (e.g. dalteparin, bemiparin, certoparin, dalteparin,
enoxaparin,
nadroparin, parnaparin, reviparin, tinzaparin), oligosaccharide (e.g.
fondaparinux,
idraparinux), heparinoid (e.g. danaparoid, dermatan sulfate, sulodexide),
dabigatran,
rivaroxaban, apixaban, edoxaban, betrixaban, darexaban, apixaban, betrixaban,
darexaban,
edoxaban, otamixaban, rivaroxaban, letaxaban, eribaxaban, hirudin,
bivalirudin, desirudin,
lepirudin, argatroban, dabigatran, efegatran, inogatran, melagatran,
ximelagatran,
antithrombin III, defibrotide, protein C (drotrecogin alfa), ramatroban, REG1,
Vitamin E,
batroxobin, hementin, direct thrombin inhibitor (e.g. hirudin), aspirin,
fibrinogen receptor
antagonists, streptokinase, urokinase and/or tissue plasminogen activator.
PATIENT & BUSINESS METHODS
Componentry to this invention are methods of selecting a cancer treatment
regime for a
subject by providing cancerous cells, obtained from this subject who has
cancer, and
exposing the cells, in an animal model (e.g. xenograft/syngenic mouse/rat)
and/or in cell
culture, to a compound(s)/composition(s) of this invention and determining if
it is effective
against those cancer cells (as evidenced by, for example, the ability of the
invention
compound(s)/composition(s) to kill the cancer cells, reduce their motility, or
reduce the rate
at which they grow or proliferate). This information can be utilized in
choosing whether to
administer this compound(s)/composition(s) to the subject and at what dosage,
optionally
wherein this decision is made by a clinical professional such as a doctor,
vet, oncologist,
pharmacist or similar. In addition, the invention encompasses methods of
monitoring the
growth/regression/stasis of cancer(s) in a subject wherein the subject has a
compound(s)/composition(s) of this invention in their body, optionally a
therapeutically
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effective amount, optionally wherein the cancer is monitored by blood testing
and/or imaged,
optionally by one or more of PET, SPECT, X-ray, CT, MRI, ultrasound,
mammography or
some other method known to those of the art, and optionally wherein this
monitoring is
performed and/or analysed by a clinical professional(s) such as a doctor, vet,
oncologist,
radiologist, pharmacist or similar, and optionally wherein this clinical
professional(s)
modifies the amount/frequency of compound(s)/composition(s) of this invention
administered
to the subject on the basis of this collected data. Componentry to this
invention are methods
of advertising/promoting/teaching/instructing/selling/distributing a
compound(s)/composition(s)/method(s) of this invention (to patient(s) and/or
clinical
professional(s)) for a therapeutic/beneficial use(s) in a subject, optionally
an anti-cancer use,
optionally wherein this method is performed by a pharmaceutical company or
subsidiary or
employee or representative.
DETAILED DESCRIPTION OF THE DRAWINGS
For purposes of clarity, not every component is labelled in every figure, nor
is every
component of each embodiment of the invention shown where illustration is not
necessary to
allow those of ordinary skill in the art to understand the invention.
Figures 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16: Experimental
evidence: molecules that
specifically inhibit the reverse mode of ATP synthase: specifically exert anti-
cancer activity:
representative, non-limiting examples.
Figures 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 show results from the NCI-60 one-
dose in vitro
assay [34-35] at the Developmental Therapeutics Program (DTP), at the National
Cancer
Institute (NCI, Bethesda, MD, USA). Its protocol is well known to those of the
art, and it
tests the effect, if any, of a test compound on the growth/survivability of a
cancer cell line as
compared to the no compound control. When this protocol was first developed, a
compound
was tested against 60 cancer cell lines, hence the name NCI-60, but more
recently this has
been reduced to 59 cell lines, and there is some variation over time in the
cancer cell lines
making up this 59 (and sometimes it drifts from being 59 also). However, a
constant is that
there is always representative cell lines from leukemia, melanoma and cancers
of the lung,
colon, brain, ovary, breast, prostate and kidney. In a one-dose NCI compound
test report, NCI
report a number for each cell line, which they call "Growth Percent", which is
its growth
relative to the no-compound control, and relative to the time zero number of
cells. This
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reported parameter allows detection of both growth inhibition (values between
0 and 100)
and lethality (values less than 0). For example, an NCI "Growth Percent" value
of 100 means
no growth inhibition. Value of 40 means 60% growth inhibition. Value of 0
means no net
growth over the course of the experiment. Value of -40 means 60% lethality.
Value of -100
means all cells are dead. I don't present NCI one-dose data in this original
format. Instead, if
the NCI "Growth Percent" value for a cell is positive, it is manipulated: [100
minus this
original NCI-60 "Growth Percent" data point], to yield the percentage "Growth
Inhibition". If
the original NCI "Growth Percent" value for a cell is negative, it is made
positive to be the
percentage of original cancer cells (at time zero) killed: "Percentage Killed"
{and in these
cases, of course, all growth has been inhibited, so percentage "Growth
Inhibition" is then
specified to be 100% for this cancer cell line}. In my one-dose figures,
"Growth Inhibition"
(0-100%) is presented on the x-axis and, if applicable, "Percentage Killed" (0-
100%) further
along on the x-axis. The latter is applicable when there is not just cancer
growth inhibition
but a reduction in the number of cancer cells from the start time i.e. when
the compound is
not merely slowing cancer growth, but is actively reducing the number of
cancer cells from
the starting number. In cases where there is only growth inhibition, only
"Growth inhibition"
is presented on the x-axis. In all cases, the greater the percentage number on
the x-axis, for a
given cancer cell line named on the y-axis, the greater the anti-cancer
activity of this
compound against this cancer cell line.
NCI-60 tests are performed at a controlled temperature of 37 C [35].
To perform the anti-cancer testing reported: BMS-199264 hydrochloride was
purchased from
Sigma-Aldrich. BTB06584 was purchased from AdooQ Bioscience, Irvine, CA, USA.
Almitrine dimesylate was purchased from Ak Scientific, Palo Alto, CA, USA.
Compounds
19a (separated into 6a and 6b stereoisomers), 31, 7a and 7b were synthesized
by reaction
schemes disclosed herein. Tested compounds are available from NCI by NSC
number which
are: BTB06584 (NSC: 794220), BMS-199264 HCl (NSC: 795767), almitrine
dimesylate
(NSC: 800450), 6h (NSC: 801828), 6a (NSC: 801827), 31 (NSC: 802605), 7b (NSC:
809247), 7a (NSC: 809248), 8a (NSC: 814041), 8b (NSC: 814040).
Figure 1: Anti-cancer activity of carboplatin in National Cancer Institute
(NCI) one-dose (10
M) assay. Data retrieved from NCI Developmental Therapeutics Program (DTP)
screening
database [32], database entry NSC: 241240. To retrieve this data, input NSC at
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https://dtp.cancer.gov/dtpstandard/dwindex/index.isp. DTP database contains
anti-cancer
performance (or more typically lack thereof) for >800,000 compounds (NSC
numbers,
wherein each NSC number represents a unique chemical entity, started at 1 and
each new
compound tested was assigned the next largest number as their NSC identifier
[I know this by
personal communication with NCI DTP] and recently assigned NSC numbers herein
are
>800,000) in the exact same experimental protocol. New entries become
publically
searchable after 3 years delay (to give time for compound submitters to secure
IP protection,
should they wish, before public disclosure by this database). This figure
presents the anti-
cancer performance of an FDA licensed cancer drug, carboplatin, one of the
most used cancer
drugs today, which is on the World Health Organisation (WHO) list of most
Essential
medicines, in exactly the same experimental protocol used for the novel cancer
drugs of this
disclosure, presented in later figures. This figure inclusion enables a direct
like-for-like
comparison between the new cancer drugs, disclosed by this disclosure, with a
cancer drug in
present, widespread clinical use. For carboplatin (10 M), mean and median
cancer growth
inhibition are 6.37% and 1.6% respectively. Maximal growth inhibition observed
is with a
leukemia cell line (SR) = 49.3%.
Figure 2: No anti-cancer activity of BTB06584 at 10 M (NCI one-dose assay).
Figure 3: Anti-cancer activity of BTB06584 at 100 M (NCI one-dose assay).
Mean and
median cancer growth inhibition are 38.76% and 33.79% respectively. Maximal
growth
inhibition observed is with a leukemia cell line (MOLT-4) = 95.38%.
Figure 4: Anti-cancer activity of BMS-199264 hydrochloride at 10 M (NCI one-
dose
assay). Mean and median cancer growth inhibition are 22.95% and 19.71%
respectively.
Maximal growth inhibition observed is with a prostate cancer cell line (PC-3)
= 85.35%.
Figure 5: Anti-cancer activity of BMS-199264 hydrochloride at 100 M (NCI one-
dose
assay).
Figure 6: Anti-cancer activity of compound 31 at 10 M (NCI one-dose assay).
Mean and
median cancer growth inhibition are 15.17% and 13.51% respectively. Maximal
growth
inhibition observed is with an NSCLC cell line (HOP-92) = 66.24%.
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Figure 7: Anti-cancer activity, in NCI one-dose assay, of almitrine dimesylate
at (7A) 10 M
and (7B) 100 M. Mean cancer growth inhibition is 32.08% (10 M) and 35.84%
(100 M).
Median cancer growth inhibition is 30.07% (10 M) and 35.32% (100 M). Maximal
growth
inhibition observed at 100 tiM is with an NSCLC cell line (HOP-92) = 76.46%
growth
inhibition. With some cancer cell lines, anti-cancer activity is less at 100
prVI than 10 RIVI
(7C).
Figure 8: (8A) A racemic mixture, or racemate, has equal amounts of the S and
R
enantiomers of a chiral molecule. Structure 19a is a racemate and (RIS)
symbolises the chiral
carbon as R or S. Structure 6a is its R stereoisomer, structure 6b is its S
stereoisomer. The
EC50 FiFo ATP hydrolase of the racemate (19a, 0.033 M) is approximately half
as potent as
that of the isolated S stereoisomer (6b, 0.018 IA) because it contains half
as much of the S
stereoisomer per unit mass, because it also contains R stereoisomer (EC50 FiFo
ATP
hydrolase > 100 M) in a 50:50 ratio. Chiral supercritical fluid
chromatography (SFC) was
used to separate 19a into its component R and S stereoisomers, which don't and
do potently
inhibit FiFo ATP hydrolase respectively (ECso values from SMPs in [5-6]), and
two samples
of opposite >97% enantiomeric excess (ee) was achieved: termed 6a and 6b
respectively.
These were independently tested in NCI one-dose (10 piM) testing: their
results are shown in
Figures (8B) and (8C) respectively. The anti-cancer activity of 6a and 6b was
similar
(Pearson correlation: R = 0.8, significant at p <0.00001). This is because
during the 48 hours
of NCI one-dose testing, they underwent racemization and their ee eroded. Such
that both
samples ultimately contained a significant proportion of S stereoisomer and
both exerted anti-
cancer activity by inhibiting FIFO ATP hydrolysis. Racemization is not
instantaneous and so
one sample, 6b, conferred greater/longer S stereoisomer exposure to the cancer
cells than the
other sample, 6a. Racemization isn't necessarily complete at testing end.
These features
explain why the 6b origin (>97% ee 6b at start) sample has greater anti-cancer
activity than
the 6a origin (>97% ee 6a at start) sample: 66% vs. 57% mean (67% vs. 59%
median) cancer
growth inhibition, across all 59 cancer cell lines, respectively. During, and
more certainly by
end, of NCI one-dose testing, 6a and 6b have 97% > ee? 50% i.e. they are a
racemate or
scalemate. Although the fidelity of the (distinction between) 6a and 6b
samples erodes during
NCI testing, as they each converge (by epimerization) upon being 19a, I still
use the 6a and
6b terms at times in this disclosure to refer to these samples during NCI
testing. In addition,
given that during NCI testing 6a ¨> 19a, and 6b --* 19a, I use the terms 6a,
6b and 19a
interchangeably at other times during this disclosure. Racemization reduces,
and increases,
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the anti-cancer activity of 6b and 6a respectively. (8D) 6b exerts greater
anti-cancer activity
than 6a. 6b is the S stereoisomer, 6a the R stereoisomer. (8E) and (8F) are
the anti-cancer
activities of 6a and 6b respectively in the NCI one-dose (100 M) assay. In
this assay,
median cancer growth inhibition for both 6a and 6b >79%. Pearson correlation
coefficient
between 6a and 6b anti-cancer activity at 100 M = 0.9437 (p < 0.00001). With
some and
overwhelmingly same cancer cell lines, for both 6a (8G) or 6b (8H), anti-
cancer activity is
much less at 100 M than 10 M.
Figure 9: Anti-cancer potency (mean % decrease in cancer growth, as compared
to no
.. compound control, in NCI-60 one-dose testing) scales with inhibition of
(EC50) Fi Fo ATP
hydrolase, across diverse chemical structures. Mean cancer growth inhibition
values are from
data in Figures 2, 3, 4, 5, 6 and S. ECso values against ATP synthase, for BMS-
199264, 19a,
6a, 6b and 31 are from sub-mitochondrial (SMP) studies in [5-8], wherein these
EC50 values
are comparable because they are from the same research group using the same
SMP assay. A
point to note, the EC50 values against ATP synthase are from bovine studies
and the anti-
cancer data is from human cancer cell lines, so there is a species difference,
and it is assumed
here that approximately the same EC50 values apply to the human case. BTB06584
potency
information (is not an EC50) is from a whole cell study [13]. Compound 31 ECso
for CYP2C9
is from [8]. Note that the presented EC50 for BMS-199264 is for BMS-199264,
not for BMS-
199264 HCI, which is the form tested for anti-cancer activity herein.
6b and 6a anti-cancer activities are similar because of their epimerization in
biological
systems, which erodes their enantiomeric excess (ee) during NCI testing,
making them
converge upon being the racemate, 19a. Thus, during NCI testing, 6b ECso FiFo
ATP
hydrolase is not constant but in the range 0.033 M > EC50 FIFO ATP hydrolase
< 0.018 M
because EC50 FiFo ATP hydrolase ¨> 0.033 M as 6b 19a, as racemization
proceeds.
Similarly, 6a ECso FIFO ATP hydrolase 0.033 M as 6a ¨> 19a.
Mean % cancer growth inhibition for BMS-199264 at 100 M is >100% because for
most
cancer cell lines tested it doesn't just cause 100% cancer growth inhibition
but, in addition,
causes cancer regression, wherein the number of cancer cells at experiment end
is less than at
experiment start. BMS-199264 predominantly exerts anti-cancer activity at 10
M by
inhibiting FiFo ATP hydrolase, and at 100 M, by reducing FIFO ATP synthesis.
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At 10 M, 6b (and 6a) exerts more anti-cancer activity than BMS-199264,
despite having
less effect on F1 F0-ATP synthesis, because it inhibits FiFo-ATP hydrolysis
more potently.
This shows that FiFo-ATP hydrolysis is an anti-cancer target. BTB06584 doesn't
exert anti-
cancer activity at 10 M, only at 100 M, wherein it has been shown only to
inhibit Fi Fo-
ATP hydrolysis at >100 M, and doesn't inhibit FiFo-ATP synthesis at this
concentration
[13]. Reaffirming that Fi Fo-ATP hydrolysis is an anti-cancer target. 31
exerts less anti-cancer
activity than BMS-199264 at 10 M. Despite 31 having the lower EC50 for FiFo-
ATP
hydrolysis [7-8]. However, BMS-199264 could be recruiting additional anti-
cancer activity at
M by reducing F1 F0-ATP synthesis also, wherein BMS-199264 has a lower EC50
for
10 FiFo-ATP synthesis than 31 ([18 9.5 M [7], so potentially 8.5 M] vs.
>30 M [8]
respectively). Furthermore [7] reports EC50 values for pure BMS-199264 whereas
I test BMS-
199264 HC1, which might be more soluble and thence more potent, with lower
EC5ovalues
for FiFo-ATP synthesis and hydrolysis than the pure form.
31 has less anti-cancer activity than its ECso FIFO ATP hydrolase value would
predict because
it is broken down by cytochrome P450 enzyme: CYP2C9, which it inhibits
competitively (31
being consumed in the process). Average 1og2 transcript intensity of CYP2C9,
across all
NCI-60 cell lines, is 3.539 [31-32]. Average 10g2 transcript intensity of
ATP5A1, the alpha
subunit of Fi ATP synthase, across all NCI-60 lines, is 9.871 [31-32]. There
are 3 alpha
subunits per ATP synthase [1]. So, on average, approximately, there is a
comparable amount
of CYP2C9 and ATP synthase in an NCI-60 cancer cell line: 3.539:(9.871/3=3.29)
1. If we
equate ECsoas some measure of binding affinity then compound 31 has a greater
affinity for
binding ATP synthase in its reverse mode (EC5o= 0.022 M) than for binding
CYP2C9
(ECso = 2 M) (these EC50 values come from different assays, thence this
comparison isn't
very robust). However, ATP synthase does not always operate in reverse, it
likely has
different operating propensities at different stages of the cell cycle, and
compound 31 EC50
FiFo ATP synthesis is > 30 M. Thus, CYP2C9 can meaningfully reduce compound
31
inhibition of FIFO ATP hydrolase, and thence its anti-cancer activity.
Especially because
CYP2C9, a cytochrome P450 enzyme, does not merely bind and sequester compound
31, but
metabolises and inactivates compound 31 at a rate set by its kcat for compound
31.
Compound 31 as substrate for an enzyme(s) of the cytochrome P450 enzyme,
whilst itself
having anti-cancer activity, means it can add to the anti-cancer action, for
example potentiate
the anti-cancer action, of one or more other anti-cancer therapeutics which
are also broken
down by this system e.g. fluorouracil (5-FU), capecitabine (prodrug of 5-FU),
imatinib
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(gleevac), nilotinib, sorafenib, dasatinib, teniposide, tamoxifen, idarubicin
etc. N.B. CYP2C9
inhibition does not exert anti-cancer activity. Sulfaphenazole is a specific,
potent CYP2C9
inhibitor [178] and it does not exert anti-cancer activity in NCI-60 one-dose
testing (NSC
757859 in DTP database [32], 10 1iM, mean cancer growth inhibition = -3.8%,
negative
symbol signifies growth promotion (!) rather than inhibition).
Compound 31 is a 1,4-benzodiazepine, which is a compound class known to bind
albumin in
blood extensively. For example, 99% of diazepam is protein bound in the blood,
wherein the
overwhelming majority of this protein is albumin. Albumin can bind many things
non-
specifically but it has high affinity binding sites with higher affinity
binding to certain
substances [ 1 72], for example, 1,4-benzodiazepines. Indeed, albumin has two
specific high
affinity binding sites, one of which is called the "benzodiazepine site", also
called Site II, the
diazepam site or the indole-BDZ site, which can bind a range of
benzodiazepines, and so
possibly compound 31 also. Compound 31 sequestration by binding albumin in the
5% Fetal
Bovine Serum (FBS) component to NCI-60 testing medium [35] might be
contributory to the
anti-cancer underperformance of 31, less than predicted by its ECso FIR) ATP
hydrolysis.
Indeed, when 31 is tested at 10 M, there are more albumin molecules than 31
molecules in
the NCI-60 test medium (200 I medium, 5% FBS [23 mg/ml albumin on average]).
Reducing FBS amount to 2% and assaying if compound 31 exerts greater anti-
cancer activity
vs. no drug control (also with FBS reduced to 2%) would reveal how much of a
factor, if at
all, albumin drug sequesteration is for compound 31 in NCI testing.
Figure 10: Chiral supercritical fluid chromatography (SFC) was used to
separate the shown
racemate into its component R and S stereo isomers and two samples of opposite
>97%
enantiomeric excess (ee) was achieved: termed 7a and 7b respectively. 7a and
7b differ from
6a and 6b in Figure 8 because they have deuterium instead of hydrogen upon
their chiral
carbon. 7a and 7b were independently tested in NCI one-dose (10 M) testing:
their results
are shown in Figures (10B) and (10C) respectively. The anti-cancer activity of
7a and 7b
against the different cancer cell lines of the NCI-60 assay was correlated
i.e. the greater, and
lesser, of their anti-cancer activity was against the same cell lines (Pearson
correlation: R =
0.8, significant at p < 0.00001). The anti-cancer activity of compounds 6a,
6b, 7a, 7b are
correlated, which suggests that they all exert anti-cancer activity by the
same mechanism,
inhibition of FiFo ATP hydrolysis (refer Figure 8 and its legend), and the
following table
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shows their pairwise Pearson correlation (R) coefficients, all significant at
p < 0.00001,
except 7a vs. 6b, wherein p = 0.00002, significant at p <0.00003.
6a 6h
7a 0.7693 0.5205
7b 0.6614 0.6451
The following table compares the mean and median cancer growth inhibition
caused by 6a
and 6b, from Figure 8, with that of 7a and 7b from the present figure:
6a 6b (6b-6a) 7a 7b (7b-7a) (7b-7a)/(6b-
6a)
Mean 57.3 66.15 8.85 52.35 72.81 20.46 2.3
Median 58.62 66.9 8.28 52.09 74.51 22.42 2.7
7b exerts greater anti-cancer activity than 6b, 7a exerts less anti-cancer
activity than 6a.
Thence the difference between the anti-cancer activity of 7b and 7a is greater
than that
between 6b and 6a. This is because, whereas 6a and 6b have hydrogen attached
to the chiral
carbon, 7a and 7b have deuterium attached to the chiral carbon. Deuterium
slows the
racemization rate by a Kinetic Isotope Effect (KIE). So, 7b maintains its
enantiomeric excess
and anti-cancer activity better, because of a slower epimerization rate to
inactive 7a. Thence
7b exerts greater anti-cancer activity than 6b. 7a maintains its enantiomeric
excess and anti-
cancer inactivity better, because of a slower epimerization rate to active 7b.
Thence 7a exerts
less anti-cancer activity than 6a. The disparity in anti-cancer activity
between 7b and 7a is 2-3
times greater than that between 6b and 6a, which is the correct order of
magnitude for a KIE
(between 1 and 7 [179], can be greater if tunnelling is very mechanistically
relevant [180]).
(10D) Recasts data from Figures 10B and 10C. Null hypothesis: equal (0.5)
probability that
x-axis value is positive (+ve) or negative (-ve): that 7a or 7b is the more
powerful
stereoisomer for any given cell line. Binomial probability of observed number
of +ve and -ve
<0.00000001. P value (one tailed) <0.000001, Significant. Conclusion: 7b has
greater anti-
cancer activity than 7a at 10 M. 7b is the S stereoisomer, 7a the R
stereoisomer.
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Replacing hydrogen(s) with deuterium(s) on a drug structure can decrease its
efficacy, for
example replacing CH3 with CD3 on morphine decreases its analgesic potency
(ED50) and
lethality (LD50) in mice [181]. With 19 hydrogens on the structure of 6b,
there are 219
(=524,288) different deuterium/hydrogen substitution combinations possible for
6a or 6b. It
isn't predictable which of these will decrease, increase or be neutral to any
anti-cancer
activity, and/or whether a given substitution(s) will impact any anti-cancer
activity of 6a and
6b in the same direction or not, at what amplitude for each, if at all. Before
this experiment
the anti-cancer impact, if any, of deuterating 6a and 6b on their chiral
carbon, to produce 7a
and 7b, couldn't have been predicted. The literature shows that KIEs can be
positive [180],
negative (inverse KIE) [182] or zero (neutral) [183], wherein it can't be
predicted which will
apply in each new case. The magnitude and direction of KlEs are unpredictable.
Moreover,
the literature teaches that the epimerization of 6a and 6b is irrelevant
because [Pl, P2, P3],
using analogy (in the unpredictable arts) to the macrolide inhibitors of [14],
teaches that FiFo
ATPase inhibitors exert anti-cancer activity by inhibiting FIFO ATP synthesis,
wherein 6a and
6b are both poor to non-inhibitors of this mode (EC5o> 100 M for both [5-61),
and so both
are taught to have poor to no anti-cancer activity, wherein this anti-cancer
action, even if
present, isn't cancer selective (normal cells rely upon FIR) ATP synthesis)
and thus isn't
clinically useful. Thus interconversion between 6a and 6b is taught
irrelevant. Furthermore,
the literature teaches that their epimerization rate is slow (refer Figure 30
legend herein),
indeed data disclosed herein also teaches this (Figure 30), so their
epimerization is taught
inconsequential for this reason also. However, novel data disclosed by this
invention
(Figures 8 & 23) shows that 6a and 6b rapidy epimerize in biological systems.
By the
invention of the present disclosure, 6b, the S stereoisomer, is discovered to
selectively exert
anti-cancer activity by selectively inhibiting FIFO ATP hydrolysis (reverse
mode), wherein
6a, the R stereoisomer, only exerts anti-cancer activity by epimerizing to
become 6b, the S
stereoisomer. That 7b has greater anti-cancer activity than 6b, and 7a has
less anti-cancer
activity than 6a, is further evidence (adding to that of Figure 9) that 6b
exerts anti-cancer
activity by inhibiting FIFO ATP hydrolysis and further teaches that, by the
most fundamental
invention of this disclosure, anti-cancer activity can be specifically exerted
by specifically
inhibiting FiFo ATP hydrolysis. A new composition of matter, 7b, is taught by
the present
disclosure and, furthermore, a method of its use for anti-cancer therapy. It
wasn't known
prior to this invention that 6b would exert greater anti-cancer activity than
6a, that 7b would
exert greater anti-cancer activity than 6b, nor that 7a would exert less anti-
cancer activity
than 6a.
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Deuteration increases and decreases the anti-cancer activity of the active, S.
and inactive, R,
stereoisomers respectively. So, to acquire greater net anti-cancer activity
from this effect, the
S stereoisomer must be administered in enantiomeric excess to the R
stereoisomer. In some
embodiments of this invention, 7b {or a pharmaceutically-acceptable salt,
solvate, hydrate or
prodrug thereof} is administered in enantiomeric excess (ee) to 7a {or a
pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof} (non-limiting examples:
ee = >70%, ee =
>95%, >99% more preferred, =100% most preferred) for use in a method of
treatment of the
human or animal body by therapy, optionally to treat/ameliorate/prevent/combat
one or more
diseases/disorders/conditions referred to in this disclosure, optionally
cancer, optionally for
the treatment/amelioration/prevention/combat of cancer in a subject(s), and/or
for the
manufacture of a medicament, optionally for treating one or more of the
diseases/disorders/conditions referred to in this disclosure, optionally
cancer. The greater the
% deuterium enrichment at the chiral carbon of 7b, and the greater the
enantiomeric excess of
7b, the more preferred the embodiment.
Figure 11: Chiral supercritical fluid chromatography (SFC) was used to
separate the shown
racemate into its component Rand S stereoisomers and two samples of opposite
>97%
enantiomeric excess (ee) was achieved: termed 8a and 8b respectively. 8a and
8b differ from
6a and 6b in Figure 8 because they have methyl (Me, CH3), instead of hydrogen
(H), upon
their chiral carbon. 8a and 8b were independently tested in NCI one-dose (10
M) testing:
their results are shown in Figures (11B) and (11C) respectively. The anti-
cancer activity of 8a
and 8b against the different cancer cell lines of the NCI-60 assay was
correlated i.e. the
greater, and lesser, of their anti-cancer activity was against the same cell
lines (Pearson
correlation: R = 0.5669, significant at p <0.00001). This correlation is
notably less than for
6a vs. 6b (0.7991) and 7a vs. 7b (0.8049).
(11D) Recasts data from Figures 11B and 11C. Null hypothesis: equal (0.5)
probability that
x-axis value is positive (+ve) or negative (-ye): that 8a or 8b is the more
powerful
stereoisomer for any given cell line. Binomial probability of observed number
of +ve (45)
and ¨ve (15) {n=60} = 0.00004613852. P value (one tailed) for 45 or more +ve
(n = 60) =
0.000091. Conclusion: 8b has greater anti-cancer activity than 8a at 10 M.
But one can see
that there are notable exceptions, which are discussed now with 11E.
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(11E) 8a and 8b NCI one-dose (10 M) test results side by side. % cancer
growth inhibition
> 100 means there are less cancer cells at experiment end than beginning (i.e.
cancer killing
activity), = 200 = all cancer cells dead at experiment end. The observed
pattern of 8a vs. 8b
activity is hard to explain. One would expect one stereoisomer to have greater
or equal
.. activity than the other. Whereas here, 8b mostly has greater activity, and
in some cases much
greater (e.g. A=85.34% with MDA-MB-231/ATCC), but in some cases 8a has the
much
greater activity (e.g. A=38.57 with A498). Moreover, for one cancer cell line
(NCI-H322M),
8a has no activity, yet 8b does. While not wishing to be bound by theory, the
following model
can explain these results, wherein the anti-cancer activity of 8a and 8b is
set by (1) the
cancer's sensitivity to a specific FIF0 ATP hydrolysis (over synthesis)
inhibitor, which can
vary between different cancer cell lines, and (2) the activity of an enzyme(s)
that hydroxilates
the methyl group (CH3) of 8a and 8b to CH2OH, which can vary between different
cancer cell
lines, wherein this enzyme(s) is referred to herein as CYP, but it needn't
necessarily be a
Cytochrome P450 enzyme(s) as other hydroxylase/monooxygenase enzymes are known
to
those of the art:
Hydroxylase/
=¨=N rµ__.1,1 OH
CH3N 0 monooxygenase N 0
CI
enzyme(s) CI
N NH N NH
CI CI
CI CI
Moreover, the ability to exert specific FIFO ATP hydrolysis inhibition, and
thence anti-cancer
activity, is ranked: R (CH20H) > S (CH2OH) S (Me) > R (Me), wherein RI S
refers to
stereochemistry and the group in brackets is the group on the chiral carbon.
If CYP activity is
low or non-existant, R (Me) and S (Me) are the predominant intracellular
species of 6a and 6b
respectively, and so, for anti-cancer activity, 6a < 6h. If CYP activity is
high, R (CH2OH) and
S(CH2OH) are the predominant intracellular species of 6a and 6b respectively,
and so, for
anti-cancer activity, 6a> 6b. Is hard to delinearate whether S (CH2OH) or S
(Me) has the
greater FiFo ATP hydrolysis inhibitory, and thence anti-cancer, activity. S
(Me) does have
anti-cancer activity, as observed with MDA-MB-231/ATCC, when 6a activity is
much lower
than 6b and so CYP activity must be low, thence S (Me) predominates. Yet S
(CH2OH) does
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have anti-cancer activity because 6b can still exert anti-cancer activity when
6a can. The
NCI-H322M cancer cell line, against which 6a has no anti-cancer activity, may
have a
mutation in, and/or especially low expression of, the CYP enzyme(s), wherein
this prevents it
from hydroxylating the methyl of R (Me) and S (Me).
(11F) can explain this model more. 6a, R (H), and 6b, S (H), cannot and can
potently inhibit
FiFo ATP hydrolysis respectively [5-6]. This figure shows that S (Me) and S
(CH2OH) are
likely to be very structurally similar to S (H), and thence likely to also
potently inhibit FiFo
ATP hydrolysis, and to exert anti-cancer activity. Crucially, R (Me) and R
(CH2OH) are not
likely to be very structurally similar to R (H), nor each other, and in the
case of R (CH2OH)
this enables it to potently inhibit FiFo ATP hydrolysis and exert anti-cancer
activity. So, in
this (CH2OH) case, and in the priming CH3 on chiral carbon case, R over S
stereochemistry
can, in some embodiments, be favoured/desirable.
CYP activity may also be relevant to the observed anti-cancer activity of 6a
and 6b wherein
the following reaction may occur inside the cell:
-;:34 N
Hydroxylase/ N
N 0 monooxygenase
OH N N 0
CI H A enzyme(s) CI
Cl
N NH N NH ______________ N NH
CI ci ci
ci ci ci
So, 6a and 6b may converge upon the same structure inside the cell, which may
explain
partially how they can both inhibit FIFO ATP hydrolysis and exert anti-cancer
activity,
because they both converge upon a structure more similar to that of S (H) than
R (H), wherein
the 2"d step is likely very fast, the 1St step might be slower, set by the
amount of enzyme(s)
available, which could allow S (H) and R (H) to endure for a period, which
could explain how
S (H) can exert greater anti-cancer activity than R (H), when they both
ultimately arrive at the
same product. The equivalent scheme may apply for 7a and 7b but wherein the
conversion is
slower because the CD bond is stronger and harder to break than a CH bond
(Kinetic Isotope
Effect, KIE), which means S (D) and R (D) endure for longer and the anti-
cancer activity
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disparity between 7a and 7b is greater than between 6a and 6b, wherein such
disparity is
experimentally observed herein.
Figure 12: The anti-cancer activity of compounds 6a, 6b, 7a, 7b, 8a and 8b are
all correlated,
which suggests that they all exert anti-cancer activity by the same mechanism,
inhibition of
F1F0 ATP hydrolysis (refer Figure 8 and its legend), and the bottom table
shows their
pairwise Pearson correlation (R) coefficients, all significant (p<0.05). In
some invention
embodiments, one or more of the compounds shown in this figure is in
stereoisomeric excess
such that it rotates polarized light in the levorotatory (L) direction,
optionally wherein this
compound(s) in stereoisomeric excess (L-rotating) is used to convey therapy in
a subject, in a
method of treatment of the human or animal body by therapy, optionally to
treat/ameliorate/prevent/combat one or more diseases/disorders/conditions
referred to in this
disclosure, optionally cancer, optionally for the
treatment/amelioration/prevention/combat of
cancer in a subject(s), and/or for the manufacture of a medicament, optionally
for treating one
or more of the diseases/disorders/conditions referred to in this disclosure,
optionally cancer.
Without wishing to be bound by theory, the order of anti-cancer activity can
be explained if
the order of inhibitory potency against F1F0 ATP hydrolysis is: R (CH2OH) > S
(CH2OH) S
(Me) > S (D) > S (H) > R (H) > R (D) > R (Me). Explaining first that 6a, R
(H), and 6b, S (H),
cannot and can potently inhibit FIFO ATP hydrolysis respectively [5-6]. Then,
explaining
from the bottom, R (D) has greater anti-cancer activity than R (Me) because it
has some
chance of racemizing to S (D) or S (H). R (H) has greater anti-cancer activity
than R (D)
because a CH bond is weaker than a CD bond and so it has a greater chance of
racemizing to
S (D) or S (H). S (D) has greater anti-cancer activity than S (H) because a CD
bond is stronger
than a CH bond and so it has less chance of racemizing to R (D) or R (H). S
(Me) and S
(CH2OH) have greater anti-cancer activity than S (D) and S (H) because they
have no (but
non-zero) chance of racemizing to R (H). R (CH2OH) has a structure very suited
to inhibiting
FiFo ATP hydrolysis, very distinct from the structure of R (H), more akin to S
(H), but better.
Below, the stereoisomer excess of the 6a, 6b, 7a, 7b, 8a and 8b samples used
in this study, as
determined by a number of different methods. For the primary data and
calculations behind
the numbers in this table, refer Figures 32, 33, 35 and their legends.
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STEREOISOMER EXCESS
UV chromatogram Chiral SFC-MS Chiral SFC
Alternative name herein Absolute(%) ee(%) Absolute(%) ee(%) Absolute(%) ee(%)
6a Stereoisomer 2 98.83 97.66 99.64 99.28
6b Stereoisomer 1 98.23 96.46 97.9 95.8
7a Stereoisomer B 98.71 97.42
7b Stereoisomer A 100 100
8a Stereoisomer 100 100 99.57 99.14 100 100
8b Stereoisomer a 100 100 99.89 99.78 100 100
Enantiomeric excess (ee) = (Absolute excess-SO) *2
Optical rotation data:
[a]= a/(c*L)
where [a] = specific rotation ( )
a = observed rotation (0)
c = concentration (g/m1)
L = length of polarimeter tube (in decimetres (dm))
Instrument used: RUDOLPH AUTOPOL V. Following were
each performed with small amount of sample. So, whilst the
direction of optical rotation is likely accurate, the magnitude
of rotation is likely not. Positive rotation = Dextrorotatory (0).
6a
1 g/100m1= 0.01 g/ml, diluted with Me0H,
20 C, A = 298 nm,
[a].38 rim= 438 nm/(c*L) = 0.075/(0.01*0.5) = +150
7a
0.25 g/100m1= 0.0025 g/ml, diluted with CHCI3, 25 C,
= 589 nm, Optical rotation = positive (+)
7b
0.25 g/100m1= 0.0025 g/ml, diluted with CHCI3, 25 C,
= 589 nm, Optical rotation = negative (-)
8a
1.0270 g/100m1= 0.01027 g/ml, diluted with Me0H,
25 C, A.= D = sodium D line = 589 nm, two trials:
[a]g= aL5/(c*L) = 0.023/(0.01027*0.5) = 4.48
= 0.024/(0.01027*0.5) = 4.67
mean = +4.58
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Figure 13: Anti-cancer activity of BMS-199264 hydrochloride. Results are from
the NCI-60
five-dose in vitro assay [34-35] at the Developmental Therapeutics Program
(DTP), at the
National Cancer Institute (NCI, Bethesda, MD, USA). In this assay, which is
well known to
those of the art, a compound is tested, in vitro, against 59 different cancer
cell lines, sourced
from 9 different tissue types, across 5 different concentrations. 9 graphs in
9 sub-figures are
presented (labelled 13A to 131), one for each of the 9 tissue types. These
graphs are as
outputted by the NCI but changed from colour to black and white. On the y-axis
of each is
the aforementioned "Growth Percent" parameter used by NCI, which is growth
relative to the
no-compound control, and relative to the time zero number of cells. This
parameter allows
detection of both growth inhibition (values between 0 and 100) and lethality
(values less than
0). GI50 is the compound concentration that causes 50% growth inhibition of a
cell line
relative to the no-compound control. Each cancer cell line has a GI50 value
and the "mean
GI50" of all 59 cell lines can be calculated: this mean GI50 for BMS-199264
hydrochloride
is 3.9 M.
Figure 14: Anti-cancer activity of compound 31 in NCI-60 five-dose in vitro
assay [34-35].
Mean GI50 is 13.2 M.
Using the COMPARE algorithm [28-29], which employs a Pearson correlation
coefficient,
one observes that the NCI-60 5-dose pattern response (GI50 values) of compound
31 is
correlated to that of BMS-199264 (0.544, p <0.00001), which indicates that
they have the
same mechanism of anti-cancer action, because [30] found that the COMPARE
algorithm can
successfully group different FDA-approved anti-cancer drugs by their method of
action.
Using this COMPARE algorithm BMS-199264 and 31 are both uncorrelated to
oligomycin A
(0.009 and -0.009 respectively), which indicates that they both have a
different mechanism of
anti-cancer action to oligomycin A. This is because, unlike oligomycin A which
potently
inhibits FIN ATP synthesis and hydrolysis, there are BMS-199264 and 31
concentrations
that selectively reduce FiFo ATP hydrolysis, which exerts anti-cancer
activity, whilst leaving
FiF0 ATP synthesis intact.
Figure 15: Anti-cancer activity of 6a and 6b in NCI-60 five-dose in vitro
assay [34-35].
Mean GI50 for 6b is 0.446 M. Mean GI50 for 6a is 0.666 M. (15J) GI50 values
tend to be
lower with 6b than 6a.
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Using the COMPARE algorithm [28-29], the NCI-60 5-dose pattern responses of 6a
and 6b
are correlated (0.754, p < 0.00001). In turn, they are correlated to the
pattern response of
BMS-199264 (6a, 0.378, p = 0.004831) and 31 (6a, 0.303, p = 0.024535). But not
too
strongly. Because all these (BMS-199264, 31, 6a, 6b) potently inhibit the
reverse mode of
.. ATP synthase (EC5o< 1 M for all [5-8]; 6a inhibits by epimerising to 6b)
but BMS-199264
and 31, distinctly from 6a and 6b, reduce FIFO ATP synthesis also with an ECso
of 18 M [7]
and >30 tiN4 [8] respectively, whereas for 6a and 6b: ECso FIE) ATP synthesis
> 100 M [5-
6], so outside the concentration range tested. Thus, within the 5 doses
tested, BMS-199264
and 31 selectively reduce FiFo ATP hydrolysis at lower test concentrations and
FiF0 ATP
synthesis also at higher test concentrations (especially at 100 M, -4 on x-
axis) whereas 6a
and 6b selectively reduce FIFO ATP hydrolysis, without significantly affecting
FiFo ATP
synthesis, at all tested concentrations. So, within the 5-dose NCI assay, BMS-
199264 and 31
exert anti-cancer activity by the same mechanism as 6a and 6b at low (< 10 M)
concentrations, but by a different mechanism at high concentration (100 M).
The x-axis of the graphs of Figure 15 might be incorrect. At present, the 5
doses shown are
from -9 to -5. When the NCI first returned the 5-dose results back for 6a and
6b, the 5 doses
shown were from -7 to -3. However, this struck me as wrong and I queried this
with the NCI
and they then sent the results through a 2"d time but with different labels on
the x-axis, as
shown herein, wherein the 5 doses shown are from -9 to -5. However, I think
this might be
wrong also. I think it should be from -8 to -4, as is typical for most of the
compounds tested
in 5-dose testing in the DTP database [16]. In which case, the mean G150 for
6b and 6a is
4.46 and 6.66 M respectively. I have emailed the NCI for further
clarification on this but no
response has been forthcoming to date. But either way, this 5-dose data shows
that 6a and 6b
exerts good anti-cancer activity. This ambiguity in the x-axis of this figure
drives ambiguity
in the x-axis of figures that use this data: Figures 17 and 18.
This paragraph (alone) assumes that the x-axis of this figure is mislabelled
and makes the
aforementioned adjustment. The Pearson correlation coefficient between 6a
activity in 1-dose
(10 M) and at 10 M in 5-dose NCI-60 testing: R= 0.8943, p<0.00001. The
Pearson
correlation coefficient between 6b activity in 1-dose (10 M) and at 10 M in
5-dose NCI-60
testing: R= 0.8737, p<0.00001. In 5-dose NCI-60 testing, at 10 M, the anti-
cancer activity of
6a and 6b is greater than at 10 M in 1-dose NCI-60 testing. Mean % cancer
growth
inhibition at 10 M in 1-dose testing: 6a (57.30%), 6b (66.15%); in 5-dose
testing: 6a
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(67.61%), 6b (76.08%). Median % cancer growth inhibition at 10 M in 1-dose
testing: 6a
(58.62%), 6b (66.90%); in 5-dose testing: 6a (67%), 6b (77%). A compound
exerting greater
anti-cancer activity at 10 M in 5-dose than 1-dose NCI-60 testing is
typically what is
observed for compounds in the DTP database [16]. So, this isn't unexpected.
What is
unexpected here, to one of the art, is that, both in 1-dose and 5-dose NCI-60
testing, anti-
cancer activity is less against some cancer cell lines at 100 M than at 10
M, wherein this
effect is more pronounced in 5-dose than 1-dose testing, wherein anti-cancer
activity is
actually greater at 10 M in 5-dose than 1-dose testing. Indeed, in 5-dose
testing the mean
and median cancer growth inhibition is actually less at 100 NI than at 10 M.
Mean %
cancer growth inhibition at 100 M in 1-dose testing: 6a (70.60%), 6b
(74.94%); in 5-dose
testing: 6a (66.22%), 6h (72.42%). Median % cancer growth inhibition at 100 M
in 1-dose
testing: 6a (79.13%), 6b (82.22%); in 5-dose testing: 6a (66.22%), 6b (72%).
The Pearson
correlation coefficient between 6a activity in 1-dose (100 M) and at 100 M
in 5-dose NCI-
60 testing: R=0.0952, p=0.473232. The Pearson correlation coefficient between
6b activity in
1-dose (100 M) and at 100 M in 5-dose NCI-60 testing: R=0.1757, p=0.183162.
Here, the
non-correlation between 5-dose and 1-dose NCI-60 results at 100 M contrasts
strongly with
their very strong correlation at 10 M reported prior.
Figure 16: Anti-cancer activity of 8a and 8b in NCI-60 five-dose in vitro
assay [34-35].
Mean GI50 for 8b and 8a is 3.09 and 2.85 M respectively. So, 8a has the lower
mean GI50
and so is the more potent. In the 8a case, this GI50 is lower/better than 65%
of 102 FDA
approved cancer drugs in [30]. A salt of 8a, e.g. 8a HC1, is likely to have an
even lower GI50
in NCI five-dose testing.
Using the COMPARE algorithm [28-29], the NCI-60 5-dose pattern responses of 8a
and 8b
are correlated (0.752, p < 0.00001). Further correlations with other NCI-60 5-
dose data,
found using the COMPARE algorithm, are shown below, all significant at p<0.05
except [6b
vs. 31] and [8b vs. 31].
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8b 8a 6a 6b BMS-199264 31
8b 1
8a 0.752 1
0.329 0.415 1
6b 0.402 0.339 0.754 1
BMS-199264 0.438 0.503 0.378 0.302 1
31 0.139 0.389 0.303 0.245 0.544 1
8b exerts greater anti-cancer activity than 8a in 1-dose (10 M) NCI-60
testing (Figure 11).
8a exerts greater anti-cancer activity than 8b at 10 M in 5-dose NCI-60
testing. So, the 1-
dose and 5-dose results contradict. However, there hasn't been a mix up of 8a
and 8b samples
because in 1-dose (10 M) testing, in Figures 11E and 11D, one can see that 8b
exerts much
greater activity against, for example, the MDA-MB-231/ATCC, 0VCAR-5, HCC-2998
and
NCI-H322M cell lines than 8a. In the 5-dose testing, although most GI50s are
lower with 8a,
one can see that the GI50 for these cancer cell lines is greater with 8a than
8b (Figure 16J).
So, this signature carries through.
To revisit the above again, using Figure 16K: in 1-dose (10 M) NCI-60
testing, cancer
growth inhibition (%) with 8b tends to be greater than with 8a, and 8b-8a is
positive. By
contrast, in 5-dose testing, GI50 tends to be lower for 8a than 8b, because 8a
has the more
potent anti-cancer activity, which makes GI50(8b-8a) positive. The exceptions
to this tend to
be for cancer cell lines against which 8b exerts much greater anti-cancer
activity than 8a in 1-
dose NCI-60 testing, so wherein 8b-8a is large, and then in 5-dose testing,
GI50 tends to be
lower for 8b than 8a, which makes GI50(8b-8a) negative. For these cancer cell
lines, 8b has
so much greater anti-cancer activity than 8a in 1-dose testing that despite
the increased anti-
cancer of 8a relative to 8b in 5-dose testing, it isn't sufficient to overtake
that of 8b in these
cases.
The Pearson correlation coefficient between 8a activity in 1-dose (10 M) and
at 10 M in 5-
dose NCI-60 testing: R=0.4544 p=0.00034. The Pearson correlation coefficient
between 8b
activity in 1-dose (10 M) and at 10 M in 5-dose NCI-60 testing: R=0.6156,
p<0.00001. In
5-dose NCI-60 testing, at 10 NI, the anti-cancer activity of 8a and 8b is
greater than at
10 M in 1-dose NCI-60 testing. Mean % cancer growth inhibition at 10 M in 1-
dose
testing: 8a (60.32%), 8b (76.51%); in 5-dose testing: 8a (102.97%), 8b
(99.42%). Median %
cancer growth inhibition at 10 M in 1-dose testing: 8a (59.83%), 8b (76.99%);
in 5-dose
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testing: 8a (95%), 8b (93%). A compound exerting greater anti-cancer activity
at 10 M in 5-
dose than 1-dose NCI-60 testing is typically what is observed for compounds in
the DTP
database [16]. So, this isn't unexpected. Not wishing to be bound by theory,
in 5-dose NCI-60
testing, why 8a has a lower mean GI50 than 8b will now be explained. 8a does
exert greater
anti-cancer activity than 8b against some cancer cell lines in 1-dose (10 M)
NCI-60 testing
(Figures 11D and 11E). As explained in the legend of Figure 11, because it can
be
metabolized to a form (hydroxylated on the chiral carbon) with greater anti-
cancer activity
than 8b. In 5-dose NCI-60 testing, whatever the reason(s) that compounds tend
to exert
greater anti-cancer activity at 10 M in 5-dose than 1-dose NCI-60 testing,
perhaps (to
speculate) relating to greater compound availability because better
care/optimization is taken
over compound solubilizing, means there is more 8a, so more 8a being
metabolized to the
more active form (increased substrate, increased reaction rate and product),
and this tips 8a
into exerting greater anti-cancer activity at 10 M than 8b. Except for the
cancer cell lines
that 8b exerted much more activity than 8a at in 1-dose testing, wherein the
increased activity
of 8a can't completely make up the deficit and overtake the activity of 8b.
Figure 17: Greater anti-cancer activity occurs with greater anti-cancer drug
concentration.
This is what someone of the art would expect. However, compounds 6a and 6b
can, upon
some (and overwhelmingly same) cancer cell lines, exert less anti-cancer
activity at 100 M
than 10 M in the NCI one-dose assay (Figures 17A and 17B) and can, upon some
(and
overwhelmingly same) cancer cell lines, exert less anti-cancer activity at 10
M than 1 M in
the NCI five-dose assay (Figure 17C). In Figures 17A and 17B, most inhibited
cancer cell
lines at 10 M are least inhibited at 100 M. Next most inhibited cancer cell
lines at 10 M
undergo cell death at 100 M. Least inhibited cancer cell lines at 10 M are
more inhibited at
100 M. Thus, to interpret, as one increases compound concentration, there are
3 zones
passed, with their boundaries different for different cancer cell lines: (1)
increased compound
concentration increases anti-cancer activity, (2) compound causes cancer cell
death, (3)
increased compound concentration decreases anti-cancer activity. The anti-
cancer activity of
compounds 6a and 6b is highly correlated, which verifies the voracity of this
data. 6b is
.. stronger acting than 6a in the NCI one-dose assay and in NCI five-dose
assay also (lower
mean GI50), wherein 6b has more cancer cell lines with less activity at 10 M
than 1 M.
Interpretation: 6a, and 6b, [compound] increase/decrease driven
increase/decrease in anti-
cancer activity is by action on same target. Data in Figures 17A and 17B is
from data in
Figure 8. Data in Figure 17C is from data in Figure 15. Supplementary data to
Figure 17C:
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NCI-60 5-dose data:
Mean GI50: 6b=0.446 M, 6a=0.666 uM
% cancer growth inhibition:
by [6b]=1 M: Mean=76.08%, Median=77%
by [6a]=1 M: Mean=67.61%, Median=67%
by [6b]=10uM: Mean=72.42%, Median=72%
by [6a]=10pM: Mean=66.22%, Median=66%
Mb: Mean=-3.66%, Median=-4%
Ma: Mean=-2.17%, Median=-2%
Pearson R (Ma, Mb) = 0.78 Ip<0.000011
Number of "uptickers": 6b=40, 6a=32
Showing that greater incidence and amplitude of "upticking" (less anti-cancer
activity at 10
than 1 M) occurs with 6b, the more potently (lower GI50) anti-cancer
stereoisomer, which
indicates that anti-cancer and upticking activity is caused by drug action
upon the same
molecular target, which is discussed further in the legend of Figure 18.
Figure 18: This diagram is an interpretation of experimental data in Figures
8, 15 and 17. It
does not itself present real data. (18A) In Figure 15, increasing
concentration of 6a or 6b
(both symbolised as 19a in the present figure) slows cancer proliferation,
until a
concentration of maximum slowing, after which further increase in compound
concentration
decreases cancer growth inhibition. The present diagram incorporates Figure 15
data with an
additional observation from Figure 8, wherein high 6a and 6b doses decrease
the cell number
of some cancer cell lines, which I suggest is due to apoptosis. This
observation is
incorporated by a narrow dosage range that causes cancer cell death, the
boundaries of which
varies by cancer cell line, and it's narrowness explains why cancer cell
killing is not observed
for most cancer cell lines in the broadly separated doses of NCI five-dose
testing. The
diagram shows that increasing compound concentration increasingly blocks FIN
ATP
hydrolysis in cancer cells, which increasingly increases their OXPHOS rate,
and thence
reactive oxygen species EROS]. Elevated [ROS] slows proliferation by ROS
checkpoint
blockade and atrophies DNA information fidelity, which reduces the number of
possible cell
divisions from limitless to a value increasingly convergent upon the Hayflick
limit [184] of
normal cells. At the inverted peak, the OXPHOS rate is so great, and EROS] so
elevated,
apoptosis is triggered (by comparison, normal cells use this OXPHOS rate
routinely).
However, as compound concentration is increased beyond this point, greater
FIFO ATP
hydrolysis inhibition makes OXPHOS more efficient (less ATP needs to be made
because
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less ATP is hydrolysed), which reduces the OXPHOS rate and [ROS], and anti-
cancer
activity is less. (18B) Upper panel shows same diagram as (A), relating to a
cancer cell,
above an equivalent diagram for a normal cell, with equivalent x-axis. It
shows that there are
concentrations of an FIR ATP hydrolysis inhibitor(s) that harm cancer and help
normal cells.
Cancers have, and need, lower intracellular [ROS] than normal cells. There are
concentrations of FIFO ATP hydrolysis inhibitor(s) that simultaneously raise
[ROS] in cancer
cells and decrease [ROS] in normal cells. In normal cells, greater FIFO ATP
hydrolysis
inhibition makes OXPHOS more efficient (less ATP needs to be made because less
ATP is
hydrolysed), which reduces the OXPHOS rate and [ROS] and increases normal cell
lifespan.
FiFo ATP hydrolysis inhibitor(s) conferred OXPHOS efficiency gain comes from,
and so its
maximum is dictated by, proportion of OXPHOS produced ATP hydrolysed by FIFO
ATP
hydrolysis, which is high, and so lifespan (and healthspan) extension
significant, especially if
it reduces EROS] sufficiently to reduce DNA mutation rate below DNA repair
rate.
Especially, if this EROS] is sufficient to keep cells differentiated,
maintaining tissue and
organ function.
Figure 19: This diagram is an interpretation of experimental data in Figure
13. It does not
itself present real data. BMS-199264 is distinct from compound 6b because, in
addition to
inhibiting FiFo ATP hydrolysis, it can also significantly reduce FiFo ATP
synthesis at NCI
tested concentrations. Background: BMS-199264 ECso FiFo ATP hydrolysis = 0.48
p,M, ECso
FIR) ATP synthesis = 18 M. (19A) Cancer cell. At lower [BMS-199264], anti-
cancer
activity is predominantly by inhibition of Fi Fo ATP hydrolysis in cancer
cells, which
increases their OXPHOS rate, and thence reactive oxygen species [ROS].
Elevated EROS]
slows proliferation by ROS checkpoint blockade and atrophies DNA information
fidelity,
which reduces the number of possible cell divisions from limitless to a value,
at increasing
[BMS-199264], increasingly convergent upon the Hayflick limit [184] of normal
cells. At
higher [BMS-199264], anti-cancer activity is additionally by reduction of FIR)
ATP
synthesis, principally by BMS-199264 conferred uncoupling of the proton motive
force (refer
Figure 27), which increases cancer OXPHOS rate and intracellular EROS]. So
much that
apoptosis ensues. (19B) Upper panel shows same diagram as (A), relating to a
cancer cell,
above equivalent diagram for a normal cell, with equivalent x-axis. It shows
that lower
[BMS-199264], which inhibits FiFo ATP hydrolysis, harms cancer and helps
normal cells.
Thus, there is a therapeutic window. However, higher [BMS-199264], which
significantly
reduces F1F0 ATP synthesis, harms both cancer and normal cells. Greater EROS]
sensitivity
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of cancer, running an embryonic stem (ES) cell type phenotype, shown in
relative positioning
of plots upon equivalent x-axis.
Figure 20: This figure recasts data from Figure 16. Greater anti-cancer
activity occurs with
greater anti-cancer drug concentration. This is what someone of the art would
expect.
However, compounds 8a and 8b can exert LESS anti-cancer activity at 100 M
than 10 M.
Looking at the mean and median % cancer growth inhibition, 8a has greater anti-
cancer
activity at 10 M, and much less anti-cancer activity at 100 1.04, than 8b.
This is because
there is a much greater number of cancer cell lines with less activity at 100
M than 10 M
with 8a (29) than 8b (6). Indeed, whilst mean and median A8b are positive
(14.70%, 11%),
mean and median A8a are negative (-1.21%, -1%). In terminology developed
herein:
8a has greater "upticking" than 8b. 8a is the more potent stereoisomer, with a
lower mean
GI50 (2.85 [04) than 8b (3.09 M). The fact that the stereoisomer with greater
anti-cancer
potency (lower GI50), 8a, has greater upticking (less anti-cancer activity at
higher drug
concentrations), and that there is a correlation (R=0.371, p=0.04755, just
significant at
p<0.05) between anti-cancer and upticking magnitude (Figure 20B), suggests
that anti-cancer
activity and its upticking might be exerted by action upon the same molecular
target: reverse
mode of ATP synthase, wherein the mechanisms involved have been disclosed in
the legend
of Figure 18. Another possible explanation for the upticking is drug
aggregation at higher
.. concentration (discussed in legend of Figure 23), but it isn't very likely
that opposite
stereoisomers could have the large difference in drug aggregation properties
required to
account for the upticking disparity between 8a and 8b. (P.S. to remind, if %
cancer growth
inhibition > 100 = less cancer cells at experiment end than start: if = 200% =
all cancer cells
dead).
Figure 21: Greater anti-cancer activity occurs with greater anti-cancer drug
concentration.
This is what someone of the art would expect. However, almitrine dimesylate
can, upon some
(24 of 57) cancer cell lines, exert less anti-cancer activity at 100 M than
10 M in the NCI
one-dose assay. As shown in Figure 7C and this figure. Regarding ATP synthase,
almitrine
makes its forward mode less efficient (less ATP synthesized per protons
passed) and its
reverse mode more efficient (more protons pumped per ATP hydrolysed) [250-
253];
expanded upon later in this disclosure. For a cancer cell currently using
oxidative
metabolism, almitrine makes FiFo ATP synthesis less efficient, whilst
concurrently making
FIF0 ATP hydrolysis more efficient, which maintains Tim and blocks a
compensatory
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increase in OXPHOS rate to maintain [ATP], [ATP] falls, which exerts anti-
cancer activity.
Almitrine decreases the maximal ATP yield available from respiratory
substrates. For a
cancer cell currently using glycolytic metabolism, almitrine making FIFO ATP
hydrolysis
more efficient exerts anti-cancer activity because it renders less ATP
hydrolysed by FIR,
ATP hydrolysis and thence, higher cytoplasmic [ATP], greater ATP feedback
inhibition of
key glycolytic enzyme(s), slower glycolytic rate, less glycolytic
intermediates available for
biosynthesis/proliferation, slower pentose phosphate pathway (PPP) rate, less
NADPH
generated, less ROS mitigation, greater intracellular EROS], wherein ROS cause
transient
growth arrest to permanent growth arrest, to apoptosis or to necrosis,
dependent on the level
of ROS [44]. While not wishing to be bound by theory, almitrine may exert
greater anti-
cancer activity for a cancer cell predominantly and/or totally relying upon
oxidative rather
than glycolytic metabolism e.g. a cancer rigidly reliant upon glutaminolysis.
The lesser anti-
cancer activity observed at 100 M almitrine dimesylate for some cancer cell
lines might be
because the greater almitrine concentration switches them to a more glycolytic
metabolism,
wherein the anti-cancer activity exerted by almitrine is less severe for these
cancer cell lines.
Indeed, increasing inefficiency of FIFO ATP synthesis and decreasing [ATP]
should pull
through increasing glycolytic rate/metabolism, assuming glucose is available
(NCI-60 assay
is run with 2 mM L-glutamine and glucose is available from the glucose content
of the 5%
Fetal Bovine Serum (FBS) added [35], ¨0.345 mM glucose in NCI-60 asssay media
by my
.. calculation [not shown]). So, in clinical use of almitrine for anti-cancer
therapy, a lower
almitrine dose might convey greater anti-cancer therapy than a higher dose and
this invention
discloses methods (elsewhere herein) to converge closer to, or upon, an
optimal anti-cancer
dose of almitrine. Different cancer cell lines likely have different
sensitivities to almitrine
depending on their relative reliance upon oxidative versus glycolytic
metabolism (which
probably varies at different stages of their cell cycle), their malleability
between these states
and the respiratory substrate(s) available.
Delimiting to cell lines that almitrine dimesylate exerts less % cancer growth
inhibiton at 100
than 10 M (those underneath the y = 0 line, bottom panel of this figure), the
Pearson
correlation coefficient between % cancer growth inhibition at 10 M and %
cancer growth
inhibition ([@100]-[@10] M) is -0.281 (p = 0.183482). So, for almitrine
dimesylate, there is
no significant (at p < 0.05) correlation between anti-cancer activity at 10 M
and
retrenchment in anti-cancer activity at 100 M. This is in distinction to 6a
and 6b, wherein
there is (Figure 17 and legend of Figure 22), and wherein the cancer cell
lines that show
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retrenchment with 6a and 6b are less in number and distinct from those with
almitrine
dimesylate.
Figure 22A: In Figures 7, 8, 15, 16, 17, 20 and 21, less anti-cancer activity
occurs with
some cancer cell lines at 100 than 10 M drug concentration. This figure and
legend teaches
that this is not because of drug aggregation at 100 M.
In vitro, less cancer growth inhibition can occur with greater cancer drug
concentration, if
this greater concentration exceeds the drug's Critical Aggregation
Concentration (CAC) [185,
186]. The CAC of an anti-cancer drug is higher when there are more cancer
cells. Because
more cancer cells sequesters more of the drug from aggregating with itself.
Thence at a CAC
exceeding drug concentration, % cancer growth inhibition is greater, as
compared to no drug
control, the greater the number of cancer cells. So when greater drug
concentration causes
less % cancer growth inhibition, one can determine if drug aggregation is the
mechanism by
assessing whether an increased number of cancer cells causes greater % cancer
growth
inhibition as compared to no drug control. If drug aggregation isn't the
mechanism one would
conversely observe a further reduction in % cancer growth inhibition as
compared to no drug
control. This methodology is componentry to this invention.
.. I couldn't do a controlled test and change the number of cancer cells
tested against for any
given cancer cell line. Instead I cautiously use an uncontrolled methodology.
In the NCI-60
one-dose assay, different cancer cell lines have different starting cell
numbers ("innoculation
densities", reported at [187]) and different cell lines have different
proliferation rates
("doubling rates", reported at [187]). Thus, at the end of the NCI-60 assay,
which runs for 48
.. hours, different cancer cell lines have different cell numbers when no drug
is applied (no drug
control) as shown in this Figure. Different cell lines have different mean
cell diameters
(reported at [188]) and this is represented in the figure also. The following
table shows, for no
drug control, the Pearson correlation coefficient (R) between starting cell
numbers, starting
cell numbers multiplied by mean diameters (in pm), final cell numbers, final
cell numbers
multiplied by mean diameters. These all correlate so highly that only one,
final cell numbers
multiplied by mean diameters (called [N*M] herein), is shown in the figure.
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R values
Innoculation density*cell diameter final cell number final cell number*cell
diameter
Innoculation density 0.9049 0.7838
0.8368
Innoculation density*cell diameter 0.496 0.682
final cell number
0.9282
P values
Innoculation density*cell diameter final cell number final cell number*cell
diameter
Innoculation density p <0.00001 p < 0.00001 p < 0.00001
Innoculation density*cell diameter p = 0.000056 p <0.00001
final cell number p < 0.00001
IF (greater number of cancer cells, greater % cancer growth inhibition (vs. no
drug control) =
positive correlation) then {drug aggregation relevant). IF (greater number of
cancer cells,
less % cancer growth inhibition (vs. no drug control) = negative correlation)
then (drug
aggregation IRrelevant).
For almitrine dimesylate (Figure 7), the Pearson correlation coefficient
between % cancer
growth inhibition (100 M) and [N*M] is 0.1119 (p = 0.39879), between % cancer
growth
inhibition ([@100]-[@10] M) and [N*M] is 0.1282 (p = 0.341922). Delimiting to
cell lines
that almitrine dimesylate exerts less % cancer growth inhibiton at 100 than 10
M, the
Pearson correlation coefficient between % cancer growth inhibition ([@100]-
[@101 M) and
[N*M] is -0.0972 (p = 0.652062). None of these are significant at p < 0.05.
There is no
statistically significant positive correlation that could indicate drug
aggregation as a
significant factor in this almitrine dimesylate anti-cancer activity data set.
For 6b and 6a (Figure 8), the Pearson correlation coefficient between % cancer
growth
inhibition (100 M) and [N*M] is -0.3811 (p = 0.002671) and -0.3784 (p =
0.002904)
respectively. For 6b and 6a, the Pearson correlation coefficient between %
cancer growth
inhibition ([@100]-[@10] M) and [N*M] is -0.3792 (p = 0.002824) and -0.3947
(p =
0.001841) respectively. Delimiting to cell lines that 6b exerts less % cancer
growth inhibiton
at 100 than 10 M, the Pearson correlation coefficient between % cancer growth
inhibition
([@100]-[@10] M) and [N*M] is -0.6201 (p = 0.023764) i.e. in this data
subset, the more
cells a cancer cell line has with no drug control, the greater the
retrenchment in % cancer
growth inhibition when 6b concentration is concentration is increased from 10
to 100 M.
Delimiting to cell lines that 6a exerts less % cancer growth inhibiton at 100
than 10 M, the
Pearson correlation coefficient between % cancer growth inhibition ([@100]-
[@10] M) and
[N*M] is -0.5355 (p = 0.072764). Leukemia cells tend to be amongst the most
numerous
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CA 3050553 2019-07-25
when no drug is added (refer figure) and are the majority of the few cell
lines that show
retrenchment in cancer growth inhibition when 6a or 6b drug concentration is
increased from
to 100 M (Figure 8G and 8H).
5 To consolidate, the 6a and 6b data, with significant (at p < 0.05)
negative correlations noted,
suggests that the retrenchment in cancer growth inhibition observed, for some
cancer cell
lines, when 6a or 6b drug concentration is increased from 10 to 100 M (Figure
8) isn't
because of drug aggregation. Moroever, the data suggests that this
retrenchment is actually by
drug action upon the same target that the drug causes cancer growth
inhibition. Because
10 greatest retrenchment at 100 M occurs with cell lines most inhibited at
10 M. These are the
cell lines most sensitive to this drug. For 6b and 6a, the Pearson correlation
coefficient
between % cancer growth inhibition at 10 M and % cancer growth inhibition
([@1001-
[@10] M) is -0.5857 (p <0.00001) and -0.6643 (p <0.00001) respectively.
Delimiting to
cell lines that 6b exerts less % cancer growth inhibiton at 100 than 10 04,
the Pearson
correlation coefficient between % cancer growth inhibition at 10 M and %
cancer growth
inhibition ([@100]-[@10] M) is -0.3656 (p = 0.219275) and -0.6243 (p =
0.030015) if the
outlier (not seen in 6a data) cell line data point (MDA-MB-231/ATCC) is
excluded from the
set. Delimiting to cell lines that 6a exerts less % cancer growth inhibiton at
100 than 10 M,
the Pearson correlation coefficient between % cancer growth inhibition at 10
M and %
cancer growth inhibition ([@100]-[@10] VI) is -0.7986 (p = 0.001062). Indeed,
looking at
Figure 17, looking especially underneath the y = 0 line, one can visually
observe the strong
correlation between greater anti-cancer activity at 10 M and lesser anti-
cancer activity at
100 M.
Delimiting to cell lines that 6a exerts less % cancer growth inhibiton at 100
than 10 M, the
mean and median retrenchment in % cancer growth inhibiton, when concentration
is
increased from 10 to 100 M, is 43.52% and 53.3% respectively (Figure 8G).
Delimiting to
cell lines that 6b exerts less % cancer growth inhibiton at 100 than 10 M,
the mean and
median retrenchment in % cancer growth inhibiton, when concentration is
increased from 10
to 100 M, is 54.73% and 66.05% respectively (Figure 8H). So, greater
retrenchment occurs
with 6b, the stereoisomer with greater anti-cancer activity (Figure 8D),
indicating that
retrenchment occurs because of drug action on the same target that the drug
acts upon to exert
anti-cancer activity.
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In Figure 17, greater incidence and amplitude of "upticking" (less anti-cancer
activity at 10
than 1 M) occurs with 6b, the more potent (lower GI50) anti-cancer
stereoisomer, which
indicates that anti-cancer and upticking activity of 6b is caused by drug
action upon the same
molecular target. Comparing Figures 8 and 15, with both 6a and 6b, greater
anti-cancer
activity at lower dose, and greater retrenchment in anti-cancer activity at
higher dose, occurs
in 5-dose than 1-dose NCI-60 testing, which indicates that 6a and 6b exert
both anti-cancer
and upticking activity by drug action on the same molecular target, which is
discussed further
in the legend of Figure 18.
In Figure 20, the fact that the stereoisomer with greater anti-cancer potency
(lower GI50), 8a,
has greater upticking (less anti-cancer activity at higher drug
concentrations), and that there is
a correlation (R=0.371, p=0.04755, just significant at p<0.05) between anti-
cancer and
upticking magnitude (Figure 20B), suggests that anti-cancer activity and its
upticking is
exerted by action upon the same molecular target: reverse mode of ATP
synthase, wherein
the mechanisms involved have been disclosed in the legend of Figure 18.
Another possible
explanation for the upticking is drug aggregation at higher concentration, but
it isn't very
likely that opposite stereoisomers could have the large difference in drug
aggregation
properties required to account for the especially large upticking disparity
observed between
8a and 8b in Figure 20.
Figure 22B: Drug aggregation assay, using methodology of [185, 186], for
compound 7b. 7b
starts to significantly aggregate (colloidal aggregation) at concentrations
higher than its
Critical Aggregation Concentration (CAC), 3.9 NI, above which it starts to
scatter light,
increasingly as its concentration is increased, as more and more 7b drug
aggregates are
formed. Drug aggregates of 7b can cause non-specific enzyme inhibition, as
shown by
inhibition of two very different enzymes, Malate dehydrogenase (MDH, IC50=3.1
M) and
AmpC beta lactamase (IC50=13.2 M), but wherein this inhibition is lifted when
Triton-X
(0.01%) detergent is added, which breaks up drug aggregates (effectively
raises the CAC).
Adding cells, e.g. cancer cells, will also raise the CAC as 7b then becomes
sequestered from
self-aggregating by binding to its intracellular target(s) and also probably
by, given its high
logP value, accumulating in cell membranes.
Could drug aggregation be the reason for the "uptick" observed with many
cancer cell lines
in the NCI-60 5-dose data for 6a and 6b (Figure 15) and/or 8a and 8b (Figure
16)? Wherein
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an "uptick" is when, after a dose is found that exerts anti-cancer activity, a
higher dose has
lower anti-cancer activity. 7b isn't 6a or 6b. But one could tentatively
suggest that the CAC
of 6b might be similar to 7b given their structural similarity. Although, a
word of caution, this
may well be wrong! But let us say that the CAC of 6b is ¨3.9 M. Of interest,
FDA approved
cancer drugs with a lower CAC value (greater propensity to aggregate), wherein
all these
CAC values are comparable as they are deduced from the same methodology [185,
186],
have been observed to have much less, or no, "upticking" in NCI-60 5-dose
tests than 6b
[32]. This might be because they have a higher affinity for their
intracellular target(s), and/or
their intracellular target(s) are more numerous, and so they are more
sequestered by the
cancer cells and their free concentration is lower in relation to their CAC.
Or, especially
because 6b might be expected to be the most sequestered by cancer cells
(membranes)
because it has the highest (computationally predicted) logP and logD values,
it might be a
sign that the "upticking" observed with 6b is not completely, or at all,
related to 6b compound
aggregation, but to a different process, for example, the process disclosed in
the legend of
Figure 18.
CAC ( M) Uptick observed in NCI-60 5-dose data? NSC number in DTP database
logP logD (pH 7)
Crizotinib 19.3 Yes (with 3 cell lines, from 109100 M) 756645 3.57
0.67
Lapatinib 0.6 Yes (with 4 cell lines, from 109100 p.M) 745750 4.64
4.19
Nilotinib 0.9 No 747599 5.36
5.33
Sorafenib 3.5 Yes (with 1 cell line, from 109100 M) 747971 4.34
4.34
Vemurafenib 1.2 No 761431 4.62
4.62
7b 3.9 ND 809247 5.97
5.89
6b ND Yes (with 40 cell lines, from 1-410 M) 801828 5.97
5.89
"Uptick" = when, after a dose is found that exerts anti-cancer activity, a
higher dose has lower anti-cancer activity.
CAC values are comparable because determined by same assay.
ND = Not Determined.
6b and 7b won't be found in DTP database until 3 years after first submitted,
by DTP policy.
DTP = Developmental Therapeutics Program (at National Cancer Institute, USA).
logP and logD: computationally predicted from structure using MarvinSketch
software (ChemAxon).
Figure 23: In vivo study of compounds 6a and 6b, conducted at 22 C room
temperature.
METHODOLOGY: 4 mice were used. Some experimental details: intravenous (IV)
administration (tail vein), dosing volume = 10 l/g, solution (not
suspension), sterilised (0.22
pm filter) vortexed vehicle= 12.5% solutol, 12.5% ethanol, 75% water, IV
solutions freshly
prepared before injection except on days 22 and 39 when solutions prepared on
previous
.. dosing days were used (sub-optimal), Female Mus Muscu/us C57BL/6 strain, 6-
8 weeks old
at study start, mice sourced from Shanghai Lingchang Bio-Technology Co. Ltd,
mice were
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observed/quarantined for a few days after arrival and before dosing, ad
libitum Co6
irradiation sterilized dry granule food & reverse osmosis autoclaved water,
corn cob bedding,
polysulfone individually ventilated cage (IVC, containing up to 5 animals):
325 mm x 210
mm x 180 mm, 12 hour light/dark cycle, 40-70% humidity, 20-26 C room
temperature,
StudyDirectorTM software (Studylog Systems, Inc. CA, USA) used to
allocate/randomize
control/treatment groups, animals marked by ear coding (notch), care and use
of animals in
accordance with the regulations of the Association for Assessment and
Accreditation of
Laboratory Animal Care (AAALAC), when recording rectal temperature: care was
taken to
ensure constant depth of probe insertion across different recordings, time was
afforded for
rectal probe temperature to equilibrate with rectal temperature, and time was
afforded
between recordings for probe temperature to reset. Rectal temperature
recording experiments
were always started between 8 and 9 am.
RESULTS: (23A) 4 mice were used, and in the presented table, "Day" refers to
the number of
days since the 1st drug/vehicle dose on Day 1. (23B) Statistics comparing
[mouse weight vs.
day, for days 1 to 44] for Mouses 1, 2, 3, 4. Mean (Mouse 2,3,4) is
significantly (at p<0.05)
different from Mouse 1, the vehicle treated control. However, Mouse 4, which
is the mouse
administered the highest cumulative drug dose, is not significantly (at
p<0.05) different from
Mouse 1, the vehicle treated control. Note that naturally, any drug treatment
aside, there is
variation in the growth trajectories of different mice individuals. Mouses 1,
2, 3, 4 are
C57BL/6 female mice. The Jackson laboratory (Jax) studied the growth
trajectory of
hundreds of C57BL/6 mice [189] and the mean and standard deviation (SD1) for
their female
C57BL/6 mice, from 7-14 weeks old, is shown in the presented graph. Only for
one data
point (17.47) is the Mean weight (Mouse 2,3,4) outside of the Jax standard
deviation (SD!),
and it is still within the 2nd standard deviation (SD2) of the Jax data (Jax
mean-SD2 = 16.9)
i.e. within the observed weight range of female Jax C57BL/6 mouse weight data.
However, a
potential source of error in comparing with the Jax data is that Mouse 1,2,3,4
are between 6-8
weeks old, i.e. it isn't known exactly how old they are, at study start. For
comparison, Jax
data is presented from 7 weeks old. However, even if Jax data from 8 weeks old
is used, the
Mean weight (Mouse 2,3,4) = 17.47 data point is still within the 2nd standard
deviation
(SD2) of the Jax data (Jax mean-SD2 = 17.2). An extant disparity is that Mouse
1,2,3,4
weren't sourced from The Jackson Laboratory, but from a different supplier of
C57BL/6 mice
i.e. they weren't C57BL/6J mice, wherein the J refers to The Jackson
Laboratory. All mice in
the present study, including the no drug control mouse, Mouse 1, have
decreases as well as
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increases in body weight during the course of observation. The administered
drug (6a or 6b)
confers mouse sedation when ambient temperature is 22 C (observed, refer
later), and so then
there is a potential margin for the drug to affect body weight if this
sedation diminishes
appetite/feeding.
(23C): For each animal, the l' rectal temperature recording is typically of an
atypically high
body temperature, which is associated with the stress of being handled, which
a mouse
becomes habituated to during the course of the experiment. This handling
effect has been
reported in other rectal thermistor studies of rodents e.g. [190-191]. Mouse 3
rectal
temperature was <30 C by 45 minutes after 20 mg/kg IV injection of 6b, it died
¨210
minutes after this IV injection, whilst drinking water, "choked on water" was
reported
observation, water temperature was at room temperature (RT) = ¨22 C (so
ingestion reduces
body temperature, if it isn't already at RT). Mouse 4 rectal temperature was
<30 C by 45
minutes after 40 mg/kg IV injection of 6b, survived >6 hours, found dead the
next day, did
not survive the night. After IV of 6b: Mouse 3 and 4 exhibited hypoactivity
and tachypnea,
both signs of hypothermia [192], coinciding with their rectal temperature drop
(<30 C).
Mouse 2 exhibited hypoactivity but recovered after 30 minutes, matching its
recovery of
rectal temperature. With 6b IV administration, there is a dose-dependent drop
in rectal
temperature.
Before 6b dosing experiments, Mouse 1, 2 and 3 had all survived IV injections
of 6a. 6a
doesn't potently reduce rectal temperature like 6b: the dose-dependent rectal
temperature
reduction (with hypoactivity reported over same timescale that rectal
temperature is reduced)
that 6a can cause is because of in vivo epimerization of 6a to 6b. Similarly,
when 6b is the
administered compound, in vivo epimerization of 6b to 6a reduces the effective
dose of 6b
and chemical modifications to the 6b compound structure to prevent or slow
this
epimerization are componentry to this invention: for example, a non-limiting
example
embodiment is to replace the hydrogen on the chiral carbon of 6b with
deuterium. >40 mg/kg
doses of 6a weren't trialled because I ran out of 6a compound. Indeed, I only
had enough 6a
to dose 2 of the 3 test mice with 40 mg/kg.
Vehicle control can cause a drop in rectal temperature because of its 12.5%
ethanol content:
12.5% of 10 ul/g solution administered = 1.25 ul/g ethanol = 0.000989226 g/g =
0.99 g/kg =
1 g/kg ethanol (IV). 1.9 g/kg ethanol (intraperitoneal injection, IP) reduced
rat body
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temperature by 1.6 C (in 24.5 to 25 C ambient temperature; raising temperature
of IP
injected ethanol solution to 37 C didn't have major impact) [193]. Ideally,
future studies
should not use ethanol as a vehicle component. The problem is not its
hypOthermia, which is
safely mitigated by a higher ambient temperature [193]. But because at just a
slightly higher
ambient temperature than this, ethanol can cause hypERthermia [193]. And the
ambient
temperature that safely mitigates ethanol driven hypothermia, without causing
ethanol driven
hyperthermia, varies with the ethanol dose [193]. This [ethanol dose/ambient
temperature/hypothermia/hyperthermia/safe rectal temperature] matrix can be
mapped by
experimentation, and indeed there is much in the literature already e.g. non-
limiting
examples: [193-199], to guide the best use of ethanol as a vehicle component
in future
studies. However, this experimentation can be avoided: alternative vehicle
options, which are
not a potent drug in and of themselves, as ethanol is, are well known to those
of the art, e.g.
see [200-201]: one or more of these can be employed as an alternative. When
ethanol as
vehicle is used, the fraction of rectal temperature drop accountable to the
test drug can be
calculated by subtracting any rectal temperature drop observed just with the
ethanol
containing vehicle control (assumes that ethanol and drug induced rectal
temperature drops
are additive and not potentiating). Drug induced rectal temperature reductions
in this study,
when they occur, are dose-dependent and well in excess of any rectal
temperature drop
observed when only ethanol containing vehicle control is injected.
The presented data shows that inhibiting the reverse mode of ATP synthase
reduces body
temperature. 6b potently inhibits the reverse mode of ATP synthase (IC5o=
0.018 j.tM [5-6]),
6a does not (IC5o> 100 M [5-6]). 6b potently reduces rectal temperature, 6a
does not (it
does to a minor degree, which is evidence for in vivo epimerization of 6a to
6b, on a faster
timescale than 6a clearance). A significant reduction in body temperature is
lethal. Thence the
maximal tolerated dose (MTD) of 6b, at room temperature = ¨22 C, is lower than
the MTD
of 6a. Body temperature cannot fall below ambient temperature and so the MTD
of 6b is
increased by ensuring ambient temperature is closer to the normal mouse body
temperature,
which ensures that mouse body temperature is maintained at an acceptable
value. This brings
greater alignment between the MTD of 6a and 6b, which in the case of 6a is
very safe: LDso
>40 mg/kg (IV). This is safer than the FDA approved anti-depressants
clomipramine HCl and
imipramine HC1: LD50 (mouse, IV} of 22 mg/kg and LD50 (mouse, IV} of 27 mg/kg
respectively (Register of Toxic Effects of Chemical Substances, RTECS). Some
patients take
these drugs daily, safely, for years.
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Non-limiting example embodiments to maintain mice, or some other animal,
including
humans, at a life permissive body temperature, whilst having a compound of
this disclosure
in their body, include locating them in a temperature-controlled room or
confmement. For
example, in small animal experiments, a plant growth (e.g. Precision
Refrigerated Plant-
Growth Incubators, Thermo Fisher Scientific Inc.) or egg or veterinary or
animal intensive
care unit (ICU) incubator or similar type device. To illustrate, and not
constrain, incubators
are made by Darwin Chambers Inc. Powers Scientific Inc, Brinsea Products Ltd.
(particularly
favoured is its Vetario range e.g. Vetario S50 model), Lyon Technologies Inc.
An
embodiment is to administer a compound of this disclosure to an animal(s),
including
human(s), in a hot country, geography or climate e.g. Dubai or somewhere else
in the Middle
East, more preferably during summer when it has high daytime and night
temperatures. There
are many methods in the literature to keep rodents at elevated temperature,
easing the cold
stress they feel at typical room temperatures [62]: e.g. partially submerging
water proof
mouse cages into fish tanks, in use as water baths, heated by thermostatic
electric fish tank
heaters [204], or by heating cages with chemical reaction hand warmers [202-
203]. Such
methods, or any method with equivalent intention, when employed with an
animal(s)/human(s) with a compound of this invention in its body, is
componentry to this
invention. Adaptive heating can be employed, which adjusts the heating element
output (e.g.
an infrared lamp, or any other heating element(s)) in response to the measured
body
temperature (e.g. by rectal temperature probe or by thermal imaging, or any
other body
temperature recording device(s)), to maintain a life-permissive body
temperature, when a
compound of this invention is in the body. With a compound of this disclosure,
the need for
(and amplitude of) ambient temperature intervention is more important for
smaller than larger
animals e.g. more so for a mouse (-20 g) than a rat (-150 g). If an
experimenter has to work
with a compound of this disclosure at typical room temperature (20-25 C) then
the test
species, and individual(s), chosen should be as large as possible. All methods
of maintaining
body temperature within a temperature range that permits life, whilst having a
compound of
this invention in the body, are componentry to this invention. For (non-
limiting) example,
.. wearing clothes.
In some invention embodiments, when the subject has a compound(s) of this
invention in its
system, for example a compound(s) of Formula (I), the subject is kept in an
ambient
temperature at or near 37 C i.e. at or near the optimal body temperature of a
mammal. This
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CA 3050553 2019-07-25
ensures that the subject's body temperature cannot fall below this optimal
body temperature.
This renders a compound(s) of this invention safer and tolerable at higher
dose, which can
enable the compound(s) to safely convey greater therapeutic utility e.g.
greater anti-cancer
activity. To illustrate by analogous example, anaesthetic can dramatically
reduce subject body
temperature, but not when the body is kept at 37 C ambient temperature [205].
An ambient
temperature equal to optimal body temperature can keep body temperature at its
optimum
when a compound(s) that reduces body heat generation and/or increases body
heat dissipation
is administered to the subject.
Figure (23D): diagram, prediction, not real experimental data. Compound 6b
reduces FIFO
ATP hydrolysis, reduces futile cycling of ATP synthesis and hydrolysis, thence
reduces
metabolic heat generation. If body temperature (BT) is greater than ambient
temperature
(AT), 6b reduces BT to be closer to AT, but it canNOT reduce BT below AT. 6b
can reduce
BT to more nearly above AT (BT=AT), not completely reducing BT=AT because
there are
other aspects to metabolic heat production than F IF ATP hydrolysis, which 6h
does not
reduce, and so BT remains higher than AT (BTzAT<BT), unless the animal dies,
in which
case BT=AT. If AT is at or above optimal body temperature, upon 6b
administration, BT will
remain at this optimum because 6b canNOT reduce BT below AT.
Figure (23E): This is a diagram relating to mouse and does NOT present real
data, although
it is inspired by experimental data in [62] and experimental data herein. When
ambient equals
thermoneutral temperature, which is ¨32 C normally for a mouse [62], the
mouse's basal
heat production (heat production of the basal metabolic rate) is sufficient to
maintain body
temperature at ¨37 C. At lower ambient temperatures than this, greater
metabolic rate/heat
production (thermogenesis) is required, and at higher ambient temperatures
than this, greater
metabolic rate is required for cooling, all to maintain body temperature at
¨37 C. A specific
FIF0 ATP hydrolysis inhibitor, e.g. compound 6b, reduces the mouse's basal
metabolic rate
and shifts its thermoneutral temperature higher, illustratively to 35 C in
this figure, which
makes the mouse more comfortable (lower metabolic rate) at higher ambient
temperatures.
Furthermore, this figure anticipates that FIE) ATP hydrolysis is integral to
thermogenic
metabolic rate, in addition to basal metabolic rate, and so the gradient of
the thermogenic
metabolic rate increase is shallower, because of reduced FIN ATP hydrolysis,
and thus the
mouse is unable to maintain 37 C body temperature at lower ambient
temperatures than its
thermoneutral temperature. The metabolic rate at thermoneutral temperature =
35 C was
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selected by drawing a line from metabolic rate at thermoneutral temperature =
32 C, = 10
W/Kg, which is an experimental data point from classic Herrington albino mouse
study
described in [62], to 37 C on the x-axis (thermoneutral temperature = 37 C,
metabolic rate =
0 W/kg) and selecting the corresponding metabolic rate for 35 C on this line.
Accordingly the
basal metabolic rate was 60% lower and, in accordance, the gradient of the
thermogenic plot
was reduced by 60% also, anticipating that FiFo ATP hydrolysis contributes
equally to basal
and thermogenic metabolic rates, although it probably contributes more to the
thermogenic
than basal metabolic rate, in which case the ascending thermogenic metabolic
gradient can be
shallower, and the descending body temperature gradient steeper, than shown
(although it
cannot outpace the gradient of ambient temperature decline: the mouse body
temperature
must be higher or at {if metabolism=0 W/kg} ambient temperature). Conclusion:
mice
administered with 6b compound can't survive at as low temperatures than
vehicle treated
mice. However, if kept at higher temperatures, at or safely greater than their
thermoneutral
temperature, the lower metabolic rate of 6b administered mice confers them
longer lifespan.
When the connection between work (unit: joule) and time is disrupted, the
simple correlation
between age and time is broken. Oxidative metabolism produces damaging/aging
reactive
oxygen species (ROS) and a lower oxidative metabolic rate produces less ROS
per unit time,
reducing the damage/aging rate, extending lifespan. An older (in time) 6b
administered body
can be younger (in aging/damage) than a younger (in time) vehicle administered
body. Like a
car, the less miles per unit time it drives, the longer it lasts, like a body
metabolism, the less
oxidative work (joules) it performs per unit time, the longer it lasts (P.S.
health benefit of
exercise doesn't breach this: exercise provokes adaptive changes that means
the body works
less at rest, e.g. endurance athletes have lower resting heart rate, = net
reduction in work
performed by body).
Figure 24: IF! is a determinant of lifespan. Background: smaller species have
a greater
metabolic rate per unit mass, faster heartbeat and shorter lifespan than
larger species [206].
Smaller species live fast, die young. Larger species live slower for longer.
Herein I disclose a
reason/mechanism for this and how to manipulate it.
Figure: data in upper two figure panels is from [207], data in lower two
panels is from the
AnAge database [208]. Interpretation here is novel. There was some margin for
error in
marrying these two data sets because [207] uses imprecise terms such as sheep,
hamster etc.
wherein there are a number of different species in [208] that can fall into
these categories. But
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CA 3050553 2019-07-25
a common sense alignment was applied in each case, by estimating which species
[207]
likely had easiest access to, so most likely used, and so most likely refer
to. So, the 12 species
that the present figure refers to are: cow (domestic cattle, Bos taurus),
mouse (house mouse,
Mus musculus), rat (Rattus rattus), hamster (golden hamster, Mesocricetus
auratus), guinea
pig (Cavia porcellus), pigeon (common wood-pigeon, Columba palumbus), chicken
(red
junglefowl, Gallus gallus), rabbit (European rabbit, Oryctolagus cuniculus),
sheep (domestic
sheep, Ovis aries), pig (wild boar, Sus scrofa), dog (Canis familiaris) and
human (Homo
sapiens). All warm blooded. Specific metabolic rate data wasn't available in
[208] for all
these species, thence the smaller number of data points in the 3rd figure
panel. The maximum
longevity of human from [208] (122.5 years) isn't shown in the figure because
arguably this
value isn't fairly comparable to the others shown, because modern medicine is
disproportionally applied to humans and the verifiable longevity data set for
humans is much,
much bigger with so many countries recording births and deaths (the bigger the
data set the
greater the chance a higher maximum longevity will be found). Human could
perhaps be
more comparably incorporated by using a maximum longevity record from a small
human
data set, to mirror the small data sets for the other species, wherein this
data set comes from
humans living in the past e.g. from 1881 Germany where life expectancy of men
and women
was 35.6 and 38.5 years respectively (Statistisches Bundesamt Deutschland,
www.destatis.de). However, omission was chosen instead. For domestic cattle
(mass=500
kg), maximum longevity of 20 years is used in my figure, which is from [208],
but [208] do
caution that this value is of "questionable" quality and say in this entry's
"observations"
section that the real value is likely to be higher, which if applied, would
make the positive
trend shown here, between body mass and lifespan, stronger.
.. The 1st figure panel shows a negative correlation between species size and
mass specific FiFo
ATP hydrolysis during ischemia. The 3rd figure panel shows a negative
correlation between
species size and mass specific metabolic rate. The 2'd figure panel shows a
negative
correlation between species size and heart rate, wherein bpm refers to beats
per minute. The
4th
4 figure panel shows a positive correlation between species size and maximum
longevity
(maximal recorded lifespan). Herein disclosed, the 1st panel correlation
drives the 3rd panel
correlation which drives the rd panel correlation which drives the 4th panel
correlation.
The following table presents the Pearson correlation coefficients, with
associated p values
below, for the different relationships.
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Pearson R correlation Coefficient
Heart rate Specific metabolic rate Maximal longevity
Specific ATPase during ischemia 0.8007 0.5331 -0.7194
Heart rate 0.9034 -0.8567
Specific metabolic rate -0.7472
P value
Heart rate Specific metabolic rate Maximal longevity
Specific ATPase during ischemia 0.001753 0.139433 0.01265
Heart rate 0.000838 0.00075
Specific metabolic rate 0.033192
IF1 inhibits FIFO ATP hydrolysis. IF I inhibits F1F0 ATP hydrolysis more
during ischemia but
its inhibition of FIR ATP hydrolysis is non-zero under normal conditions.
Larger species
inhibit specific FIFO ATP hydrolysis more than smaller species during ischemia
(1' figure
panel). This is because larger species have greater 1F1 abundance, indeed a
greater IF1/F1
ratio, and/or because their IF1 has greater inhibitory potency against F iFo
ATP hydrolysis
[207-219]. Thence there is a positive correlation between species size and
their capability to
inhibit their FIFO ATP hydrolysis i.e. the bigger the species, the less FIE)
ATP hydrolysis.
Animal mass is proportional to animal radius3, animal surface area is
proportional to animal
radius2 [220]. So, smaller animals have a larger surface area to mass ratio
and so lose a
greater proportion of their heat to the environment and so need to generate
more heat per unit
mass than larger animals, which they do by a greater metabolic rate per unit
mass, which they
achieve by greater specific FiF0 ATP hydrolysis, which consumes more ATP per
unit
mass/time, which requires more ATP be created per unit mass/time, and thence a
greater
metabolic rate, thence a greater specific heat production. So, smaller species
have greater
specific FIN ATP hydrolysis capacity than larger species (1st figure panel).
That FiF0 ATP
hydrolysis is used for heat production by animals is shown by greater FIFO ATP
hydrolysis
capability, because of greater FIFO ATP synthase and lesser IF! abundance, in
mitochondria
sourced from cows in winter than summer, wherein such seasonal changes don't
occur in
laboratory rats kept at constant temperature [209]. Furthermore, in new
experimental data
disclosed herein, specifically inhibiting FIN ATP hydrolysis in mice reduces
their heat
generation and body temperature. Because smaller species have a higher
specific metabolic
rate than larger animals, more fuel/waste is needed/ejected per unit mass per
unit time, and
they require and have a faster heart rate (2nd figure panel). Smaller species
have a greater
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specific metabolic rate (3rd panel), faster heart rate (2n(1 panel) and lower
lifespan (4th panel),
wherein I inventively suggest this is a function of their greater FIN ATP
hydrolysis capacity
(1' panel) as a function of their lesser IF' inhibitory capacity. And thence
increasing the
abundance of IFI in a species, and/or expressing/administering the IF1 amino
acid sequence
of a larger and/or longer living species, increases the lifespan of a species,
IF exogenous heat
(and/or greater body insulation) substitutes for the lower endogenous heat
production that
will ensue. Similarly, a specific/preferential drug inhibitor(s) of FIFO ATP
hydrolysis, for non-
limiting example a compound(s) of Formula (1) herein, increases the lifespan
of a subject, IF
exogenous heat (and/or greater body insulation) substitutes for the lower
endogenous heat
production that will ensue. An invention embodiment is a method of
administering a specific
or preferential inhibitor(s) of FiFo ATP hydrolysis, for non-limiting example
a compound(s)
of Formula (I-V, VII) herein, to a subject to extend their health and/or
lifespan. An invention
embodiment is a method of increasing the amount of IF! in a subject to extend
their health
and/or lifespan. An invention embodiment is a method of administering a
subject one or more
of an IF1 protein, which has a greater inhibitory potency against FIFO ATP
hydrolysis than
their endogenous IF1, especially at pH 8, to extend their health and/or
lifespan, optionally
wherein one or more of a gene or nucleotide or DNA or RNA is administered that
is
translated into such an IF! protein. An invention embodiment is to
express/administer the IF I
of a larger species in a smaller species to increase the health and/or
lifespan of the smaller
species. An invention embodiment is to express/administer the IFI of a longer
living species
in a shorter living species to increase the health and/or lifespan of the
latter.
Figure 25: In vivo, inhibiting FiFo ATP hydrolysis safely reduces the rate of
oxidative
phosphorylation and ROS generation (and thence the rate of aging). Shown in
forebrain
neurons. This figure presents re-interpreted data from [322].
[322] is an illustrative example of how an IF1 gene copy, or a mutant thereof,
from the same
or different species, can be transferred into an organism in order to increase
its IF1 protein
expression. This example shows it is safe in mouse brain (more specifically
neurons in
forebrain) to increase IF1 protein content by three times (300%), wherein the
delta increase in
[322] occurs with a mutant human IF I form with increased inhibitory potency
against FIFO
ATP hydrolysis at pH 8, which reduces FIFO ATP hydrolysis capability by ¨35%,
which
demonstrates the safety of inhibiting FiFo ATP hydrolysis in vivo, at least
specifically in
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forebrain neurons (mice were "normal in appearance, home-cage behaviour,
reproduction,
and longevity up to 1-year follow-up").
(Figure 25A) MitoSOXTM, an ROS (superoxide) reporting fluorogenic dye,
accumulates
more in the mitochondrial matrix than Tetramethylrhodamine Methyl Ester
Perchlorate
(TMRM), a fluorescent dye used to report TIM.
(Figure 25B) Experiments with mitochondria extracted from the brain of wild-
type mice (wt)
and double transgenic mice (H+/T+) with (1) a mutant human IF1 mutant gene
(H49K), with
a histidine (H) in its "pH dependence motif' {Figure 29) substituted with
lysine (K), under a
tetracycline-responsive promoter element (TRE) and (2) a tetracycline-
controlled
transactivator protein gene (tTA) under the control of the CaMKIIa promoter,
wherein
CaMKIIa is only expressed in forebrain neurons [323], thence tTA and thence
the human
H49K IF1 gene is only expressed in the mouse's forebrain neurons (in the
absence of a
.. tetracycline(s) e.g. doxycycline), wherein their IF1 amount (native +
mutant) is 3 times (i.e.
300%) greater than wild-type. H49K mutation renders IF1 with greater
inhibitory potency
against F1 F0 ATP hydrolysis at normal matrix pH (8). FiFo ATP hydrolysis is
35% less in
brain mitochondria isolated from H+/T+ than wild-type mice. The reduction in
F1F0 ATP
hydrolysis doesn't match the increase in IF1, perhaps because an IF1 fraction
is inactivated
.. by phosphorylation on its "phosphorylation control switch"/serine {Figure
29), wherein in
some invention embodiments, IF1 is used with this serine substituted for
another residue,
optionally alanine, so that it can't be phosphorylated at this position and
inactivated, and in
further embodiments, wherein this IF I also has a H49K substitution. H+/T+
mice have a
lower respiration (02 consumption, including oligomycin sensitive 02
consumption) rate than
wild-type, during State 4 (substrate [e.g. glucose, malate] stimulated) and
State 3 (+ADP
stimulated) respiration. Probably as a function of their lower respiration
rate, H+/T+ mice
have a more hyperpolarized membrane potential across their mitochondrial inner
membrane,
Pm (also referred to as Tim herein), because not so much of their proton
motive force (cant) is
being eroded per unit time to drive ATP synthesis. However, upon
FCCP/antimycin A
administration, H+/T+ mice have a more depolarized Tim than wild-type because
the
response to an uncoupler/respiratory chain inhibitor involves global reversal
of ATP synthase
and FIR, ATP hydrolysis to pump protons, partially maintaining TN, wherein
FIF0 ATP
hydrolysis capability is partially compromised in H+/T+ mice.
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CA 3050553 2019-07-25
(Figure 25C) Experiments with cortical neurons in culture, after being
cultured for 9-10
days, after being extracted from mouse embryos. The 1S and 2"d panel shows
disparity in Tim
between H+/T+ and wild-type (CRL) mice again, as in Figure 25B, but this time
in cultured
cortical neurons from the mice (embryos), wherein the mitochondria of H+/T+
mice have a
more hyperpolarized Tim (accumulate more TMRM+) than wild-type but, as in
Figure 25B,
have a more depolarized TIM upon FCCP/respiratory chain inhibitor (antimycin
A/rotenone)
administration. A typical value for TIM in normal mitochondria is -140 mV and
if we equate
the 5 a.u. value for wild-type in the 2nd panel with -140 mV, then the 6 a.u.
value of H+/T+
here in the 2"d panel is -168 mV. This Tim disparity means that H+/T+
mitochondria
.. accumulates more MitoSOX ROS (superoxide) reporting compound in their
mitochondria]
matrix, wherein this disparity can be calculated using the equation developed
in Figure 25A,
and this disparity is shown here, in the 3rd panel. With this MitoSOX
accumulation disparity,
one would expect a greater MitoSOX signal from H+/T+ mitochondria, to wild-
type
mitochondria, in the proportion shown in the 3rd panel, but what is actually
experimentally
.. observed is in the 4111 panel. Thence H+/T+ mitochondria must produce 66%
less ROS
(superoxide) than wild-type, which fits with their lower oxidative respiration
rate observed in
(Figure 25B). Thus, given that ROS are the drive to aging [325, 326], H+/T+
cells have a
slower aging rate than wild-type. Indeed, the reduced ROS in the forebrain
neurons of H+/T+
mice is likely underestimated by this in vitro cultured neuron assay. H+/T+
neurons have less
02 consumption, as shown in (Figure 25B), and so in neuron culture, because of
this lesser
02 consumption, H+/T+ neurons experience greater p02 near their respiratory
chain, which
favours increased [ROS], wherein this is an experimental artefact because in
vivo reduced 02
consumption doesn't increase p02, because breathing (rate, depth etc.)
maintains tissue p02
within a narrow range.
[322] interpret the lesser 02 consumption of H+/T+ mitochondria as evidence
that IF1
directly inhibits FIFO ATP synthesis (which suggests no clinical utility
because Fi Fo ATP
synthesis is essential to aerobic life). This is incorrect. As elucidated by
the invention of this
disclosure, substantial FIFO ATP hydrolysis is occurring under normal
conditions in mice,
which sets OXPHOS at high rate, to generate heat, as disclosed by novel
experimental data
herein (Figure 23). Increased [IF1] inhibits FIFO ATP hydrolysis more, and so
less ATP
needs to be made by FIF0 ATP synthesis, thence less OXPHOS is required, less
02 is
consumed (when Fi Fo ATP hydrolysis capability is reduced by ¨35% there is
¨60% less 02
consumption during State 3 respiration), thus less ROS are generated per unit
time, thence
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aging is slower: ¨60% slower during State 3 respiration, by the invention of
this disclosure,
reinterpreting data of [322]. Less heat is produced, but because in this case
this effect is
limited to neurons of the forebrain, heat transfer from other mouse body
regions, and from
astrocytes in the forebrain (which don't express CaMKIla, so don't express
tTA, so don't
express the transgenic IF1 gene), maintains an appropriate temperature in
forebrain neurons.
By the invention of this disclosure, the H+/T+ mice are disclosed to have
slower aging in
forebrain neurons, wherein they have reduced [susceptibility to/progression
of] brain diseases
of aging e.g. neurodegenerative diseases like Alzheimer's disease, dementia,
Parkinson's
disease etc., and less cognitive decline with aging (e.g. as assayed by one or
more of the
mouse behavioural assays disclosed elsewhere herein, or another
"brainspan"/cognitive assay
known to one of the art). This feature can be stopped by administering
tetracycline/doxycycline to these mice, which blocks IF1 transgene expression
in their
forebrain neurons. By contrast to the present invention, [322] claim that this
mutant IF1 gene
introduction increases ROS and oxidative stress in mouse forebrain neurons.
Given the
linkage between oxidative stress and the development/progression of
neurodegenerative
diseases [85], [322] teaches that these transgenic mice have increased
susceptibility
to/progression of neurodegenerative disease(s) than wild-type mice.
To briefly discuss some of the data in [322], that isn't replicated in this
figure, but that is also
.. re-interpreted herein. The protein carbonylation assay that [322] uses can
be unreliable
("numerous problems with data reproducibility or production of spurious
results" [324])
and/or, furthermore is corrupted by the large difference in [tubulin] between
the H+/T+ and
wild-type mice (refer the western blots; [tubulin] is used as a denominator in
their
carbonylation calculation). Brain of H+/T+ mice has less [ATP], but less [ADP]
also, and so
the ATP/ADP ratio isn't changed. There is greater [AMP]. These H+/T+ mice have
less ATP
and ADP because they have less cycling of FiFo ATP synthesis (ATP generation)
and FiFo
ATP hydrolysis (ADP generation), so a greater proportion of their nucleotide
exists as AMP.
This, greater [AMP], activates the AMP-activated protein kinase (AMPK; more
phosphorylated active AMPK is observed [322]), which upregulates glycolysis,
wherein the
extra NADH it produces isn't very thermodynamically inclined to enter an
electron into the
respiratory chain because of the hyperpolarised Tim (no "sink drive"), which
comes from a
reduced rate of [pmf erosion (FIFO ATP synthesis) to lesser pmf creation (FIR'
ATP
hydrolysis) to generate heat]. More thermodynamically favourable is for
lactate
dehydrogenase to use this extra NADH, switching pyruvate to lactate, which is
then exported,
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exporting this chemical energy to be accepted by a cell more in need of
energy, elsewhere in
the subject.
Figure 26: In vivo, inhibiting FiFo ATP hydrolysis safely reduces the rate of
oxidative
phosphorylation (and thence ROS generation (by extrapolation from data of
Figure 25), and
thence the rate of aging). Shown in liver cells (hepatocytes). This figure
presents re-
interpreted data from [327].
[327] is an illustrative example of how an IF I gene copy, or a mutant
thereof, from the same
.. or different species, can be safely transferred into an organism in order
to increase its IF1
protein expression. Shown in this figure is data from experiments with
mitochondria
extracted from the brain of wild-type mice (CRL) and double transgenic mice
(HIT) with (i) a
mutant human IF1 mutant gene (H49K), with a histidine (H) in its "pH
dependence motif'
{Figure 29} substituted with lysine (K), under a tetracycline-responsive
promoter element
.. (TRE) and (ii) a tetracycline-controlled transactivator protein gene (tTA)
under the control of
the rat liver-enriched activator protein (LAP; member of Cebpb gene family)
promoter,
wherein LAP is only expressed in liver cells, thence tTA and thence the human
H49K IF1
gene is only expressed in the mouse's liver cells (in the absence of a
tetracycline(s) e.g.
doxycycline). In perivenous hepatocytes especially. These are "Tet-off' mice,
expressing the
transgenic IF1 gene (h-IF1) in the absence of a tetracycline e.g. doxycycline
(Dox), as shown
in the 1' panel, wherein the 2" panel shows the presence of h-IF1 in the mouse
liver cells
using an antibody specific for human over mouse IF I , wherein this added IF1
inhibits the
FiFo ATP hydrolysis capability by 25% (31d panel) and decreases State 3
respiration rate by
37%. Alternative transgenic mice were also generated, "Tet-on" mice, which
have rtTA
instead of tTA under the control of the LAP promoter, which only express the
IF1 transgene
in the presence of a tetracycline e.g. doxycycline (Dox), wherein this added
IF I inhibits the
FiFo ATP hydrolysis capability by 40% (31d panel) and decreases State 3
respiration rate by
44%. These experiments demonstrate the safety of inhibiting FiFo ATP
hydrolysis in vivo, at
least specifically in liver (these transgenic IF1 mice had "no differences in
weight, life span
and cage behavior when compared to controls after one year of follow up").
Figure 27: In vivo, inhibiting FIFO ATP hydrolysis safely reduces the rate of
oxidative
phosphorylation (and thence ROS generation (by extrapolation from data of
Figure 25), and
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thence the rate of aging). Shown in intestine. This figure presents re-
interpreted data from
[328].
[328] is an illustrative example of how an IF1 gene copy, from the same or
different species,
can be safely transferred into an organism in order to increase its IF1
protein expression.
Shown in this figure is data from mitochondria extracted from the colon of
wild-type mice
(CL), and double transgenic mice (I/T; "Tet-on") with (i) a human IF1 (non-
mutant) gene
under a tetracycline-responsive promoter element (TRE) and with (ii) the
intestine-specific
Villin-rtTA2-M2 transactivator, thence the human IF1 gene is only expressed in
the mouse's
intestine cells (in the presence of a tetracycline(s) e.g. doxycycline). This
extra (human) IF1
inhibits the FiFo ATP hydrolysis capability by 35% and decreases oligomycin
sensitive
respiration rate by 60%. This experiment demonstrates the safety of inhibiting
FiFo ATP
hydrolysis in vivo, at least specifically in intestine.
Figure 28: Diagram (not real data) illustrating how reducing [ROS] in a cell,
for example by
inhibiting FiFo ATP hydrolysis which reduces the oxidative phosphorylation/ROS
generation
rate, can prolong/increase the information fidelity of genomic/mitochondrial
DNA, which
slows/reverses aging. ROS = reactive oxygen species. The terms in the
Michaelis-Menten
equation are extremely well known to those of the art. Figure 25 shows the
mechanism,
(inhibiting FIFO ATP hydrolysis which reduces the oxidative phosphorylation
rate) and safety
of reducing ROS in vivo. Figures 26 and 27 provide further proof of mechanism
and safety.
Figure 24 elucidates that a different rate of FIFO ATP hydrolysis is why
different species
have different maximal lifespans. Any compound(s) administered and/or
method(s) that
reduces/inhibits FiFo ATP hydrolysis to slow/reverse aging, and/or extend
lifespan/healthspan, in a subject is componentry to this invention, optionally
wherein the
expression/amount/activity of one or more DNA repair enzymes is increased in
the subject
also. For (non-limiting) example, any such use of a compound(s) of Formula I-V
and/or VII,
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof. In
some
embodiments, a compound(s) of this invention, a FiFo ATP hydrolysis
inhibitor(s), or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, is
taken/administered
before/during sleep, optionally wherein the subject sleeps in a temperature
controlled/heated
atmosphere, and/or where the subject is heated by radiative heating,
optionally wherein
exogenous heat substitutes for lower endogenous heat production by the subject
(lower
because of a compound(s) of this invention in their system), and wherein
lesser ROS
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production results, thence less DNA damage per unit time, optionally wherein
the rate of
DNA repair thence exceeds the rate of DNA damage and so there is net DNA
repair, wherein
most DNA damage can still be recognised as damage (thence the possibility of
being
repaired) within a 24 hour period, which is a factor that permits the
restorative action of sleep
-- itself, which has a metabolism slowing/body temperature dropping (ROS
reducing)
component, which a compound(s) of this invention increases/improves. In this
way, the
subject doesn't need to live in a temperature controlled environment whilst
awake, just when
they are sleeping, and/or during some other time(s) of their choosing. In some
embodiments
the subject, with a compound(s) or a pharmaceutically-acceptable salt,
solvate, hydrate or
prodrug thereof in their system, wears heat generating/retaining
clothing/device(s), optionally
that monitors the subject's body temperature and adjusts its heat
generating/retaining ability
to maintain the subject's body at or near a desired body temperature
(optionally at or near
37 C).
-- Figure 29: Some protein/peptide and nucleotide sequence embodiments of the
invention, one
or more of which, or a precursor(s)/mutant(s)/vector(s)/"expression vector(s)"
thereof, in an
aspect of the invention, are used as a medicament/in the manufacture of a
medicament/in a
pharmaceutical composition, optionally for
treating/ameliorating/preventing/combating
cancer and/or slowing aging for therapeutic (including preventative)/aesthetic
utility,
optionally wherein a vector can be a nucleotide sequence that contains a
nucleotide
sequence(s) of this invention, optionally wherein the vector is/contains a
nucleotide
sequence(s) of this invention and one or more of a promotor (constitutive or
inducible),
enhancer, inducer, termination sequence, regulatory DNA sequence, optionally
wherein the
vector is a virus or plasmid, capable or incapable of replication inside a
host cell, preferably
-- wherein the vector in a host cell leads to the expression of one or more
protein sequence
embodiments of the invention in this host cell, many applicable vectors are
known to those of
the art. Protein sequences are disclosed here using the one letter amino acid
code.
Componentry to the invention, without limitation, is a mammalian IF1, mouse IF
I, rat IF I,
rabbit IF1, guinea pig IF1, bovine IF1, canine IF1, feline IF1, non-human
primate IF! and
-- human IF I protein. (29A) These are IF1 proteins from a diversity of
species, all componentry
to the invention, as are their sub-sequences, aligned to show the incredible
conservation of
the bolded residues. The Pt bolded reside, from left to right, is the
"phosphorylation control
switch" [329] (when phosphorylated, IF I cannot bind and inhibit FiFo ATP
hydrolysis) and
the other 4 bolded residues constitute the "pH dependence motif' [330, 332,
334]
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(underlining identifies a residue that deviates from the most stereotypical
consensus
sequence). In other protein sequence embodiments of the invention, one or more
of these
bolded residues is replaced with another amino acid coded for by the genetic
code. For
example, a bolded serine (S) residue can be replaced with another residue that
can't be
phosphorylated, so that the IF1 protein cannot be inactivated by
phosphorylation at this
"phosphorylation control switch". For example, serine replaced by alanine (A).
The four
other bolded residues constitute the "pH dependence motif' and an amino acid
substitution at
one or more of its positions, optionally with lysine (K) or alanine, increases
IF1 inhibition of
FIFO ATP hydrolysis at pH 8, the normal (non-pathological) pH of the
mitochondrial matrix.
Also componentry to this invention are sub-sequences of the shown sequences
(and their
forms with bolded residue(s) substituted for another amino acid coded for by
the genetic
code) as separate stand-alone protein sequence embodiments of the invention
e.g. a sub-
sequence without the "mitochondrial import sequence/signal" (wherein this
import sequence
for human is MAVTALAARTWLGVWGVRTMQARGF [335]) or with a different
mitochondrial import sequence instead. Especially a sub-sequence with greater
inhibition of
Fi Fo ATP hydrolysis at 8. And/or a sub-sequence that aligns and
corresponds to the
"minimal inhibitory sequence" of bovine IF1, which is the smallest, minimal
sub-section of
bovine IF I that can inhibit FIFO ATP hydrolysis [330], wherein a non-limiting
candidate
"minimal inhibitory sequence" is shown in the figure, corresponding to bovine
IF1 residues
14-47 (corresponding human "minimal inhibitory sequence" is
SIREAGGAFGKREQAEEERYFRAQSREQLAALKK [335]), wherein it may actually be
shorter/longer [330] than shown (e.g. 10-47 or 16-47 or 17-47) or different
[330] (e.g.
residues 42-58 or (unlikely) 22-46 of bovine IF1). Following codes are
"primary accession
numbers" in UniProtKB database, SV = Sequence Version: (a) Q9UII2, Homo
sapiens,
SV=1, (b) H2PYG9, Pan troglodytes, SV=1, (c) G3QEV8, Gorilla gorilla gorilla,
SV=1, (d)
F6ZXX7, Macaca mulatta,SV=1, (e) A0A2U3VIM7, Odobenus rosmarus divergens,
5V=1,
(f) A0A2Y9DM04, Trichechus manatus latirostris,SV=1, (g) A9XG49, Ailuropoda
melanoleuca, SV=1, (h) E2QYN4, Canis lupus familiaris,SV=1, (i) M3WI58, Felis
catus,
SV=2, (j) F6ZXTO, Equus caballus, SV=1, (k) A0A384CECO, Ursus maritimus, SV=1,
(1)
Q03344, Rattus norvegicus, SV=2, (m) A0A2Y9LD45, Delphinapterus leucas,SV=1,
(n)
G3SWQ8, Loxodonta afi-icana,SV=1, (o) A0A2Y9EF27, Physeter catodon,SV=1, (p)
G1SEZ3, Oryctolagus cuniculus,SV=1, (q) A0A286Y43 I, Cavia porcellus, SV=1,
(r)
A0A2U3VOR3, Tursiops truncatus,SV=1, (s) A0A383Z6R7, Balaenoptera
acutorostrata
scammoni,SV=1, (t) M3YVR5, Mustela putorius furo, 5V=1, (u) Q29307, Sus
scrofa, SV=2,
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CA 3050553 2019-07-25
(v) P01096, Bos taurus, SV=2, (w) Bovine "minimal inhibitory sequence" [330,
331], (x)
G5AP86, Heterocephalus glaber, SV=1 , (y) 035143, Mus muscu/us, SV=2, (z)
S9XNE5,
Camelus ferus, SV=1, (ai) A0A1S2ZPB9, Erinaceus europaeus, SV=1, (bi)
A0A1U8CVF2,
Mesocricetus auratus, SV=1, (ci) G1NSN7, Myotis lucifugus, SV=1, (di)
A0A151PGL2,
Alligator mississippiensis,SV=1, (ei) A0A0B8RSH7, Boiga irregularis, SV=1,
(fi) H2TBT1,
Takifugu rubripes, SV=1, (gi) F7BK26, Xenopus tropicalis, SV=1, (hi)
A0A3B4D9E6,
Pygocentrus nattereri, SV=1 , (ii) A0A1D5PBD2, Gallus gallus, SV=2, (ji)
A3KNL5, Danio
rerio, SV =1 , (ki) A0A0E9WGC1, Anguilla anguilla, SV=1. One of the art will
know how to
find further IF1 sequences of other/same species, not shown here for brevity,
but which are
also componentry to this invention (as are their sub-sequences embodied as
separate stand-
alone protein embodiments of this invention, and/or are their forms with their
"phosphorylation control switch" position residue, and/or one or more of the
residues in their
"pH dependence motif', changed to another amino acid, wherein the character of
these IF I
control modules is disclosed in Figure 29B). For example, one of the art can
refer to InterPro
family "Mitochondrial ATPase inhibitor (IPR007648)" and/or Pfam family "IATP
(PF04568)". All protein sequences in the IPR007648 and/or PF04568 protein
families, and
their sub-sequences, are componentry to this invention (and their "analogs",
derivatives" and
"variants", as herein defined). Or, for example, one of the art can use the
human IF1 protein
sequence (UniProtKB: Q9U1I2, SV=1) as the query string in a "Basic Local
Alignment
Search Tool" (BLAST, https://blast.ncbi.nlm.nih.gov) search of a protein
database (protein
BLAST, BLASTp), optionally the "non-redundant protein sequences" (nr) database
at the
National Center for Biotechnology Information (NCBI) which is one of the
database options
offered with NCBI BLAST, and/or use BLAST at UniProt and search the UniProt
protein
database (https://www.uniprot.org/blast/), and from the results outputted
select protein
sequences of high BLAST score/percentage sequence similarity (in different non-
limiting
embodiments: >99%, >98%, >95%, >90%, >80%, >70%, >60%, >50%, >40%, >30%
percentage sequence similarity) to the query protein, wherein BLAST results
are returned to
the user in order of BLAST score from high to low, and/or that have one or
more IF1 motif
characteristics, such as the "pH dependence motif'. Especially preferred are
those proteins
that have been annotated in the database as an IF1 protein and/or as a protein
product of an
ATP5IF1 (alternatively known as ATPIF1) gene, wherein the human ATP5IF1 gene
has
HUGO Gene Nomenclature Committee (HGNC) ID of HGNC:871, and IF1 protein
products
of orthologs of this gene are componentry to the present invention. One of the
art knows,
and/or knows how to find, multiple online methods to find orthologs e.g. using
OrthoDB
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(www.orthodb.org, to illustrate refer to Group 1566610at2759 and/or Group
212481at40674
therein). All protein products of an ATP5IF1 gene are componentry to the
present invention.
For example, the human ATP5IFI gene has three protein products: primary
accession
numbers in UniProtKB: Q9U1I2, Q9U1I2-2 and Q9UII2-3, which are all componentry
to this
invention. When a single species has multiple IF1 proteins/homologs, e.g.
Heterocephalus
glaber has G5AP86 and A0A0P6J910 (UniProtkb primary accession numbers), e.g.
Mus
muscu/us has 035143, E9PV44 and Q8BTA7, e.g. Oryctolagus cuniculus has GI
SEZ3,
G1TES2 and GI U0F8, they are all componentry to the invention. IF1 protein
sequences (and
their "analogs", derivatives" and "variants", as herein defined), and their
sub-sequences, are
.. componentry to the present invention as are the gene and DNA and mRNA and
cDNA
sequences that can code (and the opposite complimentary base pairing strand of
the coding
sequence) for these protein sequences in a cell, wherein the genetic code,
with its redundant
character, is well known to those of the art, and wherein further nucleotide
sequences that can
hybridize with one or more of these nucleotide sequence embodiments of the
invention,
.. "under stringent conditions" (term well understood in the art, illustrative
definition included
herein), are also componentry to the present invention, as are the protein
sequences that they
encode. Given that many species have one or more of an IF1 protein sequence,
and there are
many species, there are many IF1 protein embodiments of the present invention.
Although
I Fl sequences can vary between species, they are typically functionally
interchangeable
between different species [330], wherein even a yeast IF1 sequence can inhibit
bovine FIN
ATP hydrolysis [333]. All protein sequences in the IPR007648 and/or PF04568
families of
proteins, and their sub-sequences, are componentry to this invention. As are
their "analogs"
and "derivatives" (herein defined, refer definitions section of disclosure).
As are their
variants, wherein a "variant" is a sequence of >70% sequence similarity and/or
identity, more
-- preferably >80% sequence similarity and/or identity, and increasingly
preferred: >90%,
>95%, >98%, >99% sequence similarity and/or identity. One of the art knows,
optionally
guided by software (e.g. DNASTAR Software, optionally DNASTAR Lasergene
package
[DNASTAR, Masidon Wisconsin, USA]), amino acid substitution(s) that highly
conserve
structural/chemical properties such as one or more of polarity, charge,
solubility,
hydrophobicity, hydrophilicity, amphipathic character and so can select
protein sequence
substituents, and/or variants with amino acid insertions/deletions, that
retain biological
activity, which can still inhibit FiF0 ATP hydrolysis. For protein sequence
embodiments of
this invention, post-translational modification at one or more amino acid
residues is
contemplated by, and componentry to, the invention. Illustrative post-
translational
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modifications include, without restriction, removing the initiator methionine
residue,
isomerization, methylation, alkylation, phosphorylation, glycation,
glycosylation, 0-linked
glycosylation, N-linked glycosylation, lipidation, acylation, acetylation,
amidation (for
example upon the C-terminus), deamidation, formylation, sulfation,
succinylation,
butyrylation, carbamylation, carbonylation, oxidation, biotinylation,
pegylation, gamma-
carboxylation, gamma-carboxyglutamic acid hydroxylation, pyrrolidone
carboxylic acid,
polysialylation, malonylation, hydroxylation, iodination, nucleotide addition
(e.g. ADP-
ribosylation), phosphate ester (0-linked) or phosphoramidate (N-linked)
formation,
adenylylation, uridylylation, propionylation, pyroglutamate formation, S-
glutathionylation, S-
nitrosylation, S-sulfenylation, S-sulfinylation, S-sulfonylation,
myristoylation,
palmitoylation, isoprenylation/prenylation, farnesylation,
geranilgeranilatyon, glipyatyon,
glycosylphosphatidylinositol (GPI) anchor formation, lipoylation,
phosphopantetheinylation,
one or more flavin moietys (FMN or FAD) may be covalently attached, heme C
attachment
via thioether bonds with cysteines, retinylidene Schiff base formation,
ubiquitination,
SUMOylation. Especially preferred is modification to one or more amino acids
of a protein
sequence of this invention, wherein the modification(s) is known to those of
the art to
increase the plasma stability of a protein/peptide, for example, wherein this
has been shown
with one or more protein/peptide examples in the literature. Optionally, the
protein can be
N"-methylated at one or more places. For protein sequence embodiments of this
invention, a
D-amino acid in place of the corresponding L-amino acid, at one or more
places, is
componentry to this invention. For protein sequence embodiments of this
invention,
substitution at one or more positions with a non-classical amino acid (e.g. an
amino acid not
ordinarily used by humans/mammals/eukaryotes/living systems) is contemplated
by, and
componentry to, this invention. For protein sequence embodiments of this
invention,
replacement of a negatively charged amino acid with a non-negatively charged
amino acid at
one or more positions is componentry to the invention. As is modification at
the C-terminus
to remove its negative charge, for example by amidation, or wherein at the C-
terminus,
instead of COOH, there is C(0)R or C(0)N(H)R or C(0)NR2 or R, and/or at the N
terminus
instead of NH2 there is N(H)R or NR2 or H or D or R or CH3 or C(H2)R or C(H)R2
or CR3,
wherein R is independently at each point of use selected from alkyl (e.g.
C(CH3)3),
substituted alkyl (non-limiting examples: CF3, CC13), deuterated alkyl (non-
limiting example:
CD3), aminoalkyl, thioalkyl, alkoxy, halogen, haloalkyl, haloalkoxy, any atom
or isotope
permitted by valence (including any accompanying hydrogen(s)/deuterium(s) by
valence e.g.
(non-limiting), H, NH2, SH, SiH3, PH2, BH2 etc.) including, without
limitation, La, Ti, Ce, V,
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CA 3050553 2019-07-25
Ta, Cr, Mo, Mn, Fe, Ru, Os, Co, Pd, Pt, Cu, Ag, Au, Zn, B, Al, Go, C, Si, N,
P, As, Sb, Bi, 0,
S, Se, F, Cl, Br, I, Hg. In an embodiment, one or more protein sequences of
this invention,
and/or one or more nucleotide sequences encoding one or more of such proteins,
are
administered to a subject to convey to them therapy/enhancement, optionally
cancer
treatment/amelioration/prevention/combat, optionally wherein one or more of
said nucleotide
sequences are incorporated into the subject's genome, and/or mitochondrial
DNA, in one or
more of their cells, optionally wherein the expression of this nucleotide
sequence, to protein,
is limited to a certain cell type/tissue type/organ/area/sub-section of the
subject, optionally by
the character of the promotor region incorporated with the protein(s) coding
sequence and/or
-- by where the sequence is targeted to insert into the genome and/or by where
in the subject the
nucleotide sequence (optionally in a vector) is introduced and/or by the
nature of the vector
selected. (29B) The "phosphorylation control switch" and "pH dependence motif"
of an IF1
protein [330, 332, 334]. In some invention embodiments, in an IF1 protein, the
amino acid at
the "phosphorylation control switch" is substituted with a different amino
acid coded for by
the genetic code, preferably one that can't be phosphorylated, optionally
alanine (A). And/or,
in other embodiments, one or more histidines (H), and/or tyrosine (Y), and/or
the
glutamine/glutamate, of the "pH dependence motif' is replaced with another
amino acid
coded for by the genetic code, optionally, without restriction, tyrosine (Y),
alanine (A), lysine
(K), glutamate (E), glutamine (Q), valine (V), leucine (L), isoleucine (I).
Especially preferred
is lysine (K) replacing the histidine (H) marked with a *, which corresponds
to a I-149K
(mature protein {mitochondria' import signal sequence cleaved off} numbering)
substitution
in the Bos taurus IF1 sequence. Alternatively, in alternative embodiments,
arginine (R)
replaces histidine at this position. (29C) Some protein sequences of the
invention, which are
modifications of the human IF1 protein (wherein, in further protein sequence
embodiments,
the N-terminal mitochondrial import sequence, MAVTALAARTWLGVWGVRTMQARGF,
is absent). Other IF1 protein(s), from human and/or other species, can be
modified at one or
more equivalent amino acid sequence positions, i.e. at their own
"phosphorylation control
switch" and/or in their own "pH dependence motif', by the shown rationale of
this figure,
wherein these modified protein sequences are also componentry to the present
invention, as
are nucleotide sequences that code for them by the genetic code. Any protein
sequence from
InterPro family "Mitochondrial ATPase inhibitor (IPRO07648)", and/or Pfam
family "IATP
(PF04568)", with one or more amino acid substitutions at its "phosphorylation
control
switch", and/or in its "pH dependence motif', wherein these sequence elements
are herein
defined, is componentry to the present invention. These protein embodiments,
with one or
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more post-translational modifications (examples disclosed prior), are further
embodiments of
this invention. In further embodiments of the invention, not shown in the
figure, the
substituted residue at one or more of the 5 bolded positions can be any amino
acid coded for
by the genetic code (20 standard amino acids, 21 when selenocysteine
included):
215=4,084,101 protein sequences, all componentry to the present invention (as
are their post-
translationally modified forms), as are nucleotide sequences encoding them by
the genetic
code. Further componentry to the invention are IF1 protein sequences
containing
selenomethionine and/or N-formylmethionine, optionally in place of methionine,
and/or a
"non-standard amino acid(s)" and/or a "non-proteingenic amino acid(s)" (e.g.
-- hydroxyproline) and/or a non-eukaryote amino acid(s) (e.g. pyrrolysine).
(29D) Especially
preferred is an IF1 protein with the histidine (H) marked with a * in its "pH
dependence
motif' (Figure 29B) replaced with a lysine (K). This figure shows illustrative
IF1 proteins
that have been modified at this position, wherein the site of lysine (K)
substitution is bolded,
as is the enduring remainder of the "pH dependence motif', which is
unmodified, and the
-- residue at their "phosphorylation control switch", which is unmodified.
These protein
sequences are componentry to the present invention as are other modified IF1
proteins, not
shown, which are modified in the equivalent position of their own "pH
dependence motif', as
are nucleotide sequences that code for them by the genetic code, as are sub-
sequences such as
those with their N-terminal mitochondrial import sequence absent. Any protein
sequence
from InterPro family "Mitochondrial ATPase inhibitor (IPR007648)" and/or Pfam
family
"IATP (PF04568)", with the starred (*) histidine (Figure 29B) of its "pH
dependence motif'
replaced with a lysine (K), is componentry to this invention. Illustrative IF1
proteins
modified at this position are shown in this figure (wherein the original
unmodified sequences
are also componentry to this invention): (a) Q9U1I2, Homo sapiens, SV=1, (b)
Q5RFJ9,
-- Pongo abelii,SV=1, (c) A0A2R9CQC9, Pan paniscus, SV=1, (d) A0A2J8Y5Q9,
Pongo
abelii, SV=1, (e) H2PYG9, Pan troglodytes, SV=1, (f) A0A213H 1 P7, Nomascus
leucogenys,
SV=1, (g) G3QEV8, Gorilla gorilla gorilla, SV=1, (h) A0A2K6AYV8, Macaca
nemestrina,
SV=1, (i) A0A2K5Y149, Mandrillus leucophaeus,SV=1, (j) A0A2K5POW3, Cercocebus
atys, SV=1, (k) G7NWV6, Macaca fascicularis, SV=1 , (1) A0A096NQ00, Papio
anubis,
-- SV=1, (m) F6ZXX7, Macaca mulatta,SV=1, (n) A0A0D9S814, Chlorocebus sabaeus,
SV=1, (o) A0A2K6N3T3, Rhinopithecus bieti,SV=1, (p) A0A2K6Q8H8, Rhinopithecus
roxellana, SV=1, (q) A0A2K5SG67, Cebus capucinus imitator, SV=1, (r)
A0A2K6SEK8,
Saimiri boliviensis boliviensis,SV=1, (s) A0A2K5KB15, Colobus angolensis
palliatus,
SV=1 , (t) A0A2K5DQW7, Aotus nancymaae, SV=1 , (u) F7IA10, Callithrixjacchus,
SV=1,
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(v) A0A0D9SDU9, Chlorocebus sabaeus,SV=1, (w) AOA1D5QRM5, Macaca mulatta,
SV=1, (x) A0A1U7SXJ3, Tarsius syrichta, SV=1, (y) A0A2K6Q8J3, Rhinopithecus
roxellana,SV=1, (z) A0A2K6SEL7, Saimiri boliviensis boliviensis, SV=1, (ai)
A0A2K6GYY1, Propithecus coquereli, SV=1, (bi) HOX2G2, Otolemur garnettii,SV=1,
(ci)
A0A2Y9GJM5, Neomonachus schauinslandi, SV=1, (di) A0A2K5J921, Colobus
angolensis
palliatus, SV=1 , (ei) A0A2U3VIM7, Odobenus rosmarus divergens, SV=1, (fi)
L8Y809,
Tupaia chinensis,SV=1, (gi) L5JUTO, Pteropus alecto, SV=1, (hi) A0A2Y9DM04,
Trichechus manatus latirostris,SV=1, (ii) A9XG49, Ailuropoda melanoleuca,SV=1.
(29E)
Further IF1 proteins modified to have a lysine (K) at the starred (*) position
of their "pH
dependence motif' (Figure 29B), wherein sub-sequences are componentry to the
invention,
such as those with their N-terminal mitochondrial import sequence absent, as
are the
unmodified native IF1 proteins: (a) E2QYN4, Canis lupus familiaris, SV=1, (b)
M3WIS8,
Felis catus, SV=2, (c) F6ZXTO, Equus caballus,SV=1, (d) HOXQ94, Otolemur
garnettii,
SV=1, (e) A0A384CECO, Ursus maritimus,SV=1, (f) D2GWK3, Ailuropoda
melanoleuca,
SV=1, (g) I3N8E6, ktidomys tridecemlineatus,SV=1, (h) A0A2U3YF49,
Leptonychotes
weddellii, SV=1, (i) Q03344, Rattus norvegicus, SV=2, (j) A0A341D6Q8,
Neophocaena
asiaeorientalis asiaeorientalis, SV=1, (k) A0A2Y9LD45, Delphinapterus
leucas,SV=1, (1)
G3SWQ8, Loxodonta africana,SV=1, (m) A0A2Y9EF27, Physeter catodon, SV=1, (n)
G1SEZ3, Oryctolagus cuniculus, SV=1 , (o) A0A286Y431, Cavia porcellus, SV=1,
(p)
A0A340XS26, Lipotes vexillifer, SV=1, (q) A0A2U3Y890, Leptonychotes weddellii,
SV=1,
(r) A0A1A6FZ83, Neotoma lepida,SV=1, (s) A0A2Y9J3D1, Enhydra lutris
kenyoni,SV=1,
(t) A0A2U3VOR3, Tursiops truncatus,SV=1, (u) A0A383Z6R7, Balaenoptera
acutorostrata
scammoni, SV=1, (v) M3YVR5, Mustela putorius furo,SV=1, (w) Q29307, Sus
scrofa,
SV=2, (x) L8IJ24, Bos mutus,SV=1, (y) P01096, Bos taurus, SV=2, (z)
A0A250Y8Y0,
Castor canadensis,SV=1, (ai) G5AP86, Heterocephalus glaber, SV=1, (bi) 035143,
Mus
muscu/us, SV=2, (ci) G3H1Z3, Cricetulus griseus, SV=1, (di) S9XNE5, Camelus
ferus,
SV =1 , (ei) A0A1S2ZPB9, Erinaceus europaeus, SV=1, (fi) A0A1U8CVF2,
Mesocricetus
auratus, SV=1, (gi) A0A091E4M7, Fukomys damarensis, SV=1, (hi) GIU0F8,
Oryctolagus
cuniculus, SV=1 , (ii) G1PGS1, Myotis lucifugus, SV =1, ai) F7BE70,
Monodelphis
domestica, SV=1, (ki) W5NYG6, Ovis aries, SV=1. (29F) Compare and contrast
this figure
with Figure 29B, wherein the present figure shows a scheme for especially
preferred IF1
protein sequence embodiments of this invention, which have a "phosphorylation
control
switch" that is locked in the "on" position, and an "attenuated pH dependence
motif' in place
of a "pH dependence motif'. (29G) Especially preferred is an IF1 protein with
the residue in
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its "phosphorylation control switch" position set to alanine (A) and the
starred (*) histidine
(H) of its "pH dependence motif' (Figure 29B) replaced with a lysine (K). This
figure shows
illustrative IF1 proteins that have been modified in this way. These protein
sequences are
componentry to the present invention as are other modified IF1 proteins, not
shown, which
are modified equivalently, as are nucleotide sequences that code for them by
the genetic code,
as are their protein and nucleotide sub-sequences, such as those with their N-
terminal
mitochondrial import sequence absent. Any protein sequence from InterPro
family
-Mitochondria] ATPase inhibitor (IPR007648)" and/or Pfam family "IATP
(PF04568)", with
the residue in its "phosphorylation control switch" position (Figure 29B) set
to alanine (A)
and the starred (*) histidine (Figure 29B) of its "p1-1 dependence motif'
replaced with a
lysine (K), is componentry to this invention. Illustrative IF1 proteins
modified at these two
positions are shown in this figure (wherein the original unmodified sequences
are also
componentry to this invention): (a) Q9U1I2, Homo sapiens, SV=1, (b) P01096,
Bos taurus,
SV=2, (c) 035143, Mus muscu/us, SV=2, (d) Q03344, Rattus norvegicus, SV=2, (e)
G1SEZ3, Oryctolagus cuniculus, SV=1 , (f) A0A286Y431, Cavia porcellus, SV=1 ,
(g)
E2QYN4, Canis lupus familiaris,SV=1, (h) M3WIS8, Felis catus, SV=2, (i)
F6ZXTO, Equus
caballus,SV=1, (j) G5AP86, Heterocephalus glaber, SV=1, (k) G3H1Z3, Cricetulus
griseus,
SV=1, (1) A0A1U8CVF2, Mesocricetus auratus,SV=1, (m) M3YVR5, Mustela putorius
furo, SV=1, (n) A0A2Y9LD45, Delphinapterus leucas, SV=1 , (o) G3SWQ8,
Loxodonta
africana, SV=1, (p) A0A2Y9EF27, Physeter catodon, SV=1 , (q) Q5RFJ9, Pongo
abelii,
SV=1, (r) A0A2R9CQC9, Pan paniscus, SV=1, (s) A0A2J8Y5Q9, Pongo abelii, SV=1,
(t)
H2PYG9, Pan troglodytes, SV=1, (u) A0A213H1P7, Nomascus leucogenys, SV=1 , (v)
G3QEV8, Gorilla gorilla gorilla, SV=1 , (w) A0A2K6AYV8, Macaca nemestrina,
SV=1, (x)
A0A2K5Y149, Mandrillus leucophaeus, SV=1, (y) A0A2K5POW3, Cercocebus atys,
SV=1 ,
(z) G7NWV6, Macaca fascicularis, SV=1, (ai) A0A096NQ00, Papio anubis, SV=1,
(bi)
F6ZXX7, Macaca mulatta,SV=1, (ci) A0A0D9S814, Chlorocebus sabaeus, SV=1, (di)
A0A2K6N3T3, Rhinopithecus bieti, SV=1, (ei) A0A2K6Q8H8, Rhinopithecus
roxellana,
SV=1, (fi) A0A2K5SG67, Cebus capucinus imitator, SV=1, (gi) A0A2K6SEK8,
Saimiri
boliviensis boliviensis, SV=1, (hi) A0A2K5KB15, Colobus angolensis
palliatus,SV=1, (ii)
A0A2K5DQW7, Aotus nancymaae, SV=1 (29H) Some preferred protein sequence
embodiments of the invention, applying teaching of Figure 29F to the Human IF1
protein
sequence. In further protein sequence embodiments, shown, the N-terminal
mitochondrial
import sequence, MAVTALAARTWLGVWGVRTMQARGF, is absent. (291) DNA and
protein sequence for Human IF1 protein, and a mutant thereof, wherein two
different DNA
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sequences coding for this IF1 mutant protein sequence are presented,
illustrating the
redundancy of the genetic code. In further sequence embodiments, not shown,
the N-terminal
mitochondrial import sequence is absent. (29J) Consensus DNA and protein
sequence for
Human IF I protein, and a mutant thereof. (29K) Number of DNA sequences that
code, using
the genetic code, for the Human IF1 protein, and a mutant thereof. (29L) The
human ATPIF I
gene, shown, with IF1 protein sequence coding and non-coding regions, is
componentry to
this invention, as are its sub-sequences and variants e.g. with a different
codon(s) used for
coding the same amino acid (or stop). (29M) Some protein and DNA sequence
embodiments
of the invention. The termination sequence shown is illustrative and not
restrictive. Any other
termination sequence (enables release of RNA polymerase during transcription)
can be used
in its place, wherein such sequences may comprise, without limitation, a stop
codon(s),
thymine rich region(s) (poly-T tail), cytosine rich region(s), palindromic
region(s) etc. and
wherein a termination sequence can be added at the end of any sequence of this
invention to
become a further sequence of this invention. Furthermore, a different
mitochondrial targeting
sequence than shown may be used. (29N) Some protein and DNA sequence
embodiments of
the invention. (290) Componentry to the invention is to locate a DNA
sequence(s) encoding
an IF1 protein sequence(s) of this invention in one or more mitochondria of a
subject,
wherein the DNA codons used are different (AGA and AGG are not used for
arginine (R),
one of CGT/CGC/CGA/CGG is used independently in each case instead; this figure
presents
some illustrative sequence embodiments of this invention with this
modification), because
mitochondria use a different genetic code than the nucleus, to enable the same
IF1 protein
sequence to be produced. (29P) Bovine IF1 protein, and some non-limiting sub-
sequence
embodiments of the invention, wherein all IF1 (all species) sub-sequences, or
mutant(s)
thereof, and/or concatenation thereof, are componentry to this invention, as
are the nucleotide
sequences that code for them. IF1 1-60 fragment can't dimerize and exists as a
monomer
[336], 10-46 has ten-fold less activity than 10-47 showing the importance of
the 47th residue,
14-47 has been termed the "minimal inhibitory sequence" [331, 330], 22-46 can
inhibit F1
ATP hydrolysis [337] but not F1 F0 ATP hydrolysis [338, 331]. 42-58 is an
alternative
"minimal inhibitory sequence" [338-341]. It might be that 42-58 inhibits FIR)
ATP
hydrolysis by a different binding/mechanism than 14-47 and entire IF I . Also
shown are
further non-limiting IF1 sub-sequences of this invention, which interact with
the
oligomerization residue(s) {e.g. H49} of IF1, therein blocking IF1
oligomerization, whilst not
blocking the inhibitory sequence of IF1 (which IF1 oligomerization [>dimer]
does), thus
increasing free IF1 (monomer and/or dimer) and IF1 inhibition of FIFO ATP
hydrolysis.
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CA 3050553 2019-07-25
(29Q) Sub-sequences of an IF1 protein or mutant thereof (and nucleotide
sequences that code
for them) are componentry to this invention, wherein illustrative example
embodiments are
shown. Human IF1 protein, and some non-limiting sub-sequence embodiments of
the
invention, wherein all IF1 sub-sequences, or mutant(s) thereof, and/or
concatenation thereof,
are componentry to this invention, as are the nucleotide sequences that code
for them. (29R)
A sub-sequence of an IF1 protein, or mutant thereof, in either orientation (N
to C, C to N),
attached to a mitochondrial import sequence, in either orientation, optionally
the
mitochondrial import sequence of Human IF1 (MAVTALAARTWLGVWGVRTMQARGF)
or that of a different IF1 protein, optionally that of a mammal, rodent or non-
human primate,
-- or attached to a mitochondrial import sequence of another protein that is
located to the
mitochondrial matrix, is componentry to this invention (as are the nucleotide
sequences that
code for it), wherein illustrative example embodiments are shown. (29S) An IF1
protein or
sub-sequence thereof, and/or mutant thereof, in either orientation (N to C, C
to N), attached
to a cell penetrating peptide sequence, in either orientation, optionally via
a connecting
-- glycine (increases flexibility between domains), wherein many such cell
penetrating peptide
sequences are known to those of the art (>1,800; e.g. refer to [343] for CPP
sequences, e.g.
the HIV-1 Tat cell penetrating peptide sequence, YGRKKRRQRRR), is componentry
to this
invention (as are the nucleotide sequences that code for it), wherein
illustrative example
embodiments are shown. Furthermore, a sub-sequence of an IF1 protein, or
mutant thereof, in
either orientation, attached to a mitochondrial import sequence, in either
orientation,
optionally the mitochondrial import sequence of Human IF1 (or that of a
different IF1
protein, optionally that of a mammal, rodent or non-human primate, or attached
to a
mitochondrial import sequence of another protein that is located to the
mitochondrial matrix),
which is itself attached to a cell penetrating peptide sequence (many known to
those of the
art), in either orientation, optionally via a connecting glycine, is
componentry to this
invention (as are the nucleotide sequences that code for it), wherein
illustrative example
embodiments are shown. Encompassed by the invention: the domains can be
ordered
differently that shown in the figure e.g. the "mitochondrial import sequence"
can instead be
"upstream" (closer to N terminus) of the "cell penetrating sequence" and all
possible
-- orientation (N to C, C to N) combinations are contemplated by this
invention. At all places in
this disclosure that a cell penetrating peptide, and/or CPP, is referred to,
in other invention
embodiments this is replaced/substituted with mitochondria penetrating peptide
(MPP),
wherein many such sequences are known to those of the art. To illustrate, and
not restrict,
refer to [344] or a paper/patent/patent application/document that cites it
(e.g. [345]), or a
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CA 3050553 2019-07-25
paper/patent/patent application/document that cites a paper that cites it, or
a
paper/patent/patent application/document that cites a paper that cites a paper
that cites it etc.,
optionally found using pubmed or google scholar. Or alternatively, cell
penetrating peptide,
and/or CPP, can be replaced/substituted with a non-peptide mitochondria
targeting moiety, to
illustrate (not restrict): [346-348]. (29T) An IF1 protein or sub-sequence
thereof, and/or
mutant thereof, in either orientation (N to C, C to N), attached to a cell
penetrating peptide
sequence (many known to those of the art), in either orientation, which is
attached to an
epitope tag sequence (many good tags known to those of the art), in either
orientation,
optionally wherein the cell penetrating peptide sequence is flanked by
glycines (increases
-- flexibility between domains), is componentry to this invention (as are the
nucleotide
sequences that code for it), wherein illustrative example embodiments are
shown.
Furthermore, a sub-sequence of an IF I protein, or mutant thereof, in either
orientation,
attached to a mitochondrial import sequence, in either orientation, optionally
the
mitochondrial import sequence of Human IF I (or that of a different IF1
protein, optionally
that of a mammal, rodent or non-human primate, or attached to a mitochondrial
import
sequence of another protein that is located to the mitochondrial matrix),
which is itself
attached to a cell penetrating peptide sequence (many known to those of the
art), in either
orientation, attached to an epitope tag sequence (many good tags known to
those of the art),
in either orientation, optionally wherein the cell penetrating peptide
sequence is flanked by
-- glycines, is componentry to this invention (as are the nucleotide sequences
that code for it),
wherein illustrative example embodiments are shown. Encompassed by the
invention: the
domains can be ordered differently than shown e.g. the "mitochondrial import
sequence" can
instead be "upstream" (closer to N terminus) of the "cell penetrating
sequence" and/or
"epitope tag sequence", the "cell penetrating sequence" can be "upstream" of
the "epitope tag
sequence" etc. Any IF1 protein sequence or sub-sequence or concatenation of
sub-sequences,
optionally mutant thereof, attached to a cell penetrating peptide and/or
mitochondrial import
sequence and/or epitope tag sequence, wherein all orientation (N to C or C to
N)
combinations are contemplated, is componentry to this invention, as are their
coding
nucleotide sequences. (29U) Collection of non-limiting invention embodiments.
(29V)
Collection of non-limiting invention embodiments. (29W) Some invention
embodiments:
non-limiting peptide inhibitors of FIFO ATP hydrolysis: melittin, pre-sequence
of subunit IV
of yeast cytochrome c oxidase, Syn-A2, Syn-C and A11,12 [4], bovine Fi p
subunit residues
394-413, 384-403, 404-423 [337], or mutant(s) thereof, and/or concatenation
thereof, are
componentry to this invention, as are the nucleotide sequences that code for
them.
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Encompassed by the invention: the domains can be ordered differently than
shown e.g. the
"mitochondrial import sequence" can instead be "upstream" (closer to N
terminus) of the
"cell penetrating sequence" and/or "epitope tag sequence", the "cell
penetrating sequence"
can be "upstream" of the "epitope tag sequence" etc. Any FIFO ATP hydrolysis
inhibitory
peptide or concatenation of such sequences, optionally mutant thereof,
attached to a cell
penetrating peptide and/or mitochondrial import sequence and/or epitope tag
sequence,
wherein all orientation (N to C or C to N) combinations are contemplated, is
componentry to
this invention, as are their coding nucleotide sequences. (29X) A few non-
limiting example
embodiments are shown, wherein all IF1 protein sequences and sub-sequences
from all
species, or mutant(s) thereof, and/or concatenation thereof, are componentry
to this invention,
as are the nucleotide sequences that code for them. In some embodiments one or
more of the
E in the EEE sub-sequence is replaced with an amino acid whose side-chain is
not negative,
optionally glutamine (Q) or asparagine (N), optionally a non-proteingenic
amino acid,
optionally 5,6-dehydrohomoleucine (CAS: 73322-75-5; available on
labnetwork.com e.g.
from Arena Chemical, La Mure, France) or (S)-2-amino-5-methylhexanoic acid
(CAS:
31872-98-7; available on labnetwork.com e.g. from Astatech Inc., Bristol PA,
USA). In some
embodiments, in place of one or more histidines, (S)-2-amino-3-(1H-imidazol-1-
yl)propanoic
acid (CAS 114717-14-5; PubChem CID: 12311022; available from BOC sciences,
Shirley,
NY, USA, PubChem SID: 254789149) is incorporated into the protein/peptide
chain instead.
Optionally, one or more of the NH are replaced with NCH3, especially preferred
at one or
more places on the peptide backbone i.e. one or more Nu are methylated.
Optionally, instead
of N(CH3)2 as shown here, N(CH3)3 is at the N and/or C terminal ends. Or
N(H)R, NR2, CH3,
C(H2)R, C(H)R2, CR3, R, wherein R is independently selected at each point of
use from the
options for R given earlier in this figure legend. D-amino acid in place of
the corresponding
L-amino acid, at one or more places, is componentry to this invention.
Figure 30: The stereoisomer excess (%) of 6b (and from this, by inference, 6a)
is stable in
stereoisomer excess stability experiments.
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STEREOISOMER EXCESS STABILITY of 6b sample
After 2.5 After
months 72 hours
stored as in water IN PHOSPHATE SOLUTION AT 37 C
powder @RT TIME=0
TIME=24 hours TIME=48 hours
6a fib 6a 6b 6a 6b 6a 6b 6a 6b
Retention Time (minutes) 1.68 1.91 1.67 1.88
1.858 2.125 absent 2.117 1.892 2.142
Area under peak 0.66 33.6
0 9.87 0.34 24.28
Absolute excess (%) 99.34 99.41 98.07 100
98.62
Enantiomeric excess (%) 98.68 98.82 96.14 100
97.24
Retention times can vary because of different chiral-SFC columns (of different
lengths) used.
RT=room temperature, 6b in water assay: water was in contact with air (and so
CO2), thence (by inference)
water pH ¨5.6. Phosphate buffer solution, 1 M, sourced from Sigma-Aldrich
(product code: P3619), its
recorded pH at the 37 C temperature used = 7.41. Enantiomeric excess =
(absolute excess-50)*2.
Figure 30A: 6b sample was incubated in phosphate (1 M) buffer solution at 37
C. Via chiral
SFC-MS, utilising the chiral AD-3 column (IPAJPAm_10-40_Gradient_4m1_S), the
stereoisomeric excess of 6b was recorded at the start (Time=0), after 24
hours, and after 48
hours in the phosphate solution, wherein this excess wasn't observed to be
eroded. However,
after this incubation in phosphate solution, in the 24 and 48 hour snapshots,
a new RT peak
(at RT=0.325 min) was seen, containing a molecular species at m/z 450.429, not
observed at
Time=0 (not above the level of noise; so it is not an impurity in the 6b
compound sample, nor
an impurity in the phosphate solution, nor an impurity in the chiral column
step), nor
observed after 72 hours incubation in water (data not shown), which shows that
it probably
isn't a hydrolysis product of the 6b compound. The m/z 450.429 species, and
the m/z 391.406
species, both observed in the RT=0.325 min peak, are not observed in the
RT=2.1 peak,
which contains 6b: m/z 537.08 [M+H]+. Further data for Time=0, Time=24 hours,
Time=48
-- hours incubation in phosphate solution is shown in Figures 30B, 30C and 30D
respectively.
So, this emergent RT peak likely relates to 6b compound reaction with
phosphate. Possibly
relevantly, phosphate is well known to facilitate (some mechanisms of)
racemization (e.g. "at
pH 7.4 the rate of [chiral] inversion was linearly dependent on phosphate
concentration"
[359]). Enantiomer self-disproportionation (SDE) [360] is the separation of a
non-racemic
mixture (scalemate) of enantiomers into an enantioenriched fraction and a more
racemic
fraction i.e. in an Achiral separation, when a compound has strong
intermolecular associative
forces, a racemic fraction (of predominantly hetero (R, S') oligomers e.g. R-S
dimers) can
elute separately (separate) from a fraction of enantiomeric excess (of
predominantly homo
oligomers or monomers), especially (but not restrictively) when the number of
molecules in
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CA 3050553 2019-07-25
the (most thermodynamically favored) oligomer/monomer is different. In chiral
(as opposed
to achiral) separation, such as conducted here by chiral SFC, for some
compounds (those with
significant intermolecular associative forces), there is the potential
(prediction) for 3 different
RT peaks to be observed, corresponding to the R stereoisomer (monomer or homo-
oligomer),
S stereoisomer (monomer or homo-oligomer) and racemate (hetero-oligomer). In
the
presented data here, peaks observed at RT=1.8 and RT=2.1 minutes likely relate
to the R and
S stereoisomers and, if applicable, the RT peak for the racemate is out of the
observed range
(i.e. at a higher unobserved RT) or it could be the emergent RT=0.325 min
peak, which
emerges after compound 6b incubation in phosphate solution, wherein the mono-
charged
-- (+1) form of a 6b-6a hetero-dimer, or higher oligomer, is out of the m/z
range observed
within it, wherein a multi-charged oligomer species of 6b-6a could conceivably
be within the
observed m/z range, possibly relating to the m/z 450.429 species observed.
Although,
alternatively, this emergent RT peak, with its m/z 450.429 species, could
relate to phosphate
related break up of 6b. In retrospect, what would have been helpful, which
wasn't done, is
inject 6a-6b racemate into the column to see what RT it elutes at. Then see if
this RT peak
emerges after 6b incubation in phosphate solution, wherein this would then
indicate if 6b
racemization is occurring in this experiment or not.
As disclosed elsewhere herein, 6a and 6b significantly epimerize within 48
hours in a
biological system (NCI-60 testing). This ee instability in biological media is
an unexpected
result. This finding is componentry to the invention of this disclosure, as
are any
modifications to the structure of 6a or 6b to slow their ee erosion due to
epimerization/racemization. The ee instability is unexpected because it runs
contrary to
theory. The 2nd order rate constant (kgb) for general-base catalysed
racemization can be
predicted for 6a and 6b by theory from [221] (and its supplementary
information, all
incorporated by reference). Joined to the stereogenic carbon of 6a and 6b
there is a phenyl
group, alkyl and a guanidine group: IAAG(RI,R2,R3) = -19.9 +2.8 -19.1 = -36.2
kcal/mol.
The value for the guanidine group (19.1) is not from [221] but from one of its
authors, Dr.
Andrew Leach, by personal communication, who sourced it by quantum mechanical
calculation with the Gaussian 03 software package. The cross-conjugation
correction [221]
was not applied in this case because Dr. Leach wrote "I would not expect
guanidine to be a
conjugating group". log(kg) = -0.11 * IAAG(RI,R2,R3) - 9.81, Kgb =
0.00000148593 M-Is-
1. This is not particularly fast. Not seeking to be bound by theory,
racemization might occur
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CA 3050553 2019-07-25
faster for 6a and 6b in cells than predicted by theory because their high
lipophibicity
concentrates them in biological membranes. 6a logP = 5.97, calculated from 6a
structure
using [25]. This means 6a accumulates -1,000,000 times more in model-lipid
octanol than
water, which means 6a disproportionally accumulates in biological membranes,
which drives
it to high concentration, which permits significant intermolecular
racemization. The
protonable nitrogen atom of the 6a imidazole group pulls a proton off the
stereogenic carbon
of another 6a molecule, leaving a planar carboanion, and when a proton
reattaches to this
carbon there is an equal probability of it doing so from either side, and so
there is a 50%
chance that the stereochemistry changes. Thence, racemization proceeds at a
faster rate than
predicted by methodology of [221], the present state of the art in
racemization
theory/prediction. The same process can apply to 6b. Deuterium in place of
hydrogen on the
stereogenic carbon of 6b decreases the rate of this racemization, because a
carbon-deuterium
bond is stronger than a carbon-hydrogen bond (kinetic istope effect, KIE) and
this
isotopologue of 6b is componentry to this invention. The difference in %
cancer growth
inhibition between 6a and 6b for each cell line at 10 04 drug dose (in NCI-60
one-dose
testing) correlates with the difference in % cancer growth inhibition between
6a and 6b for
each cell line at 1 1.1M drug dose (in NC1-60 five-dose testing). Pearson
correlation
coefficient, R = 0.7919, p-value <0.00001, significant at p <0.05. This shows
that there are
cellular factors, which can vary between different cancer cell lines,
determining the rate of 6a
and/or 6b racemization. For example, this could be membrane volume and/or the
level of
expression of a racemase/epimerase enzyme(s), and/or an enzyme(s) that
converts the drug
into a form that can undergo enzyme-catalysed epimerisation. In NCI-60 one-
dose testing at
10 04, across cell lines tested, there is a positive correlation between %
cancer growth
inhibition of 6b and the product [(% cancer growth inhibition due to 6b) - (%
cancer growth
inhibition due to 6a)]. Pearson correlation coefficient, R = 0.4167, p =
0.000927, significant
at p <0.05. Thus, when the anti-cancer performance of 6a is further behind the
anti-cancer
performance of 6b, anti-cancer performance of 6b is better. This suggests that
for 6a to have
greater anti-cancer activity, 6b needs to have less anti-cancer activity.
Conversely, for 6b to
have greater anti-cancer activity, 6a needs to have less anti-cancer activity.
This is because
what gives 6a its anti-cancer activity, racemization, takes away anti-cancer
activity from 6b.
Thus, in cancer cell lines that enable faster epimerization of 6a and 6b,
there is greater 6a vs.
6b anti-cancer activity. This explains why, for NCI-60 one-dose (10 04) assay,
across the cell
lines, there is a negative correlation (although not significant) between %
cancer growth
inhibition of 6a and the product [(% cancer growth inhibition due to 6b) - (%
cancer growth
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CA 3050553 2019-07-25
inhibition due to 6a)]. Pearson correlation coefficient, R = -0.2136, but not
significant at p <
0.05. The same phenomenon (6a and 6b compete for anti-cancer activity, because
what gives
to 6a, racemization, takes away from 6b) is observed in NCI60 five-dose data
at 1 M.
Across cell lines tested, there is a positive correlation between % cancer
growth inhibition of
.. 6b and the product [(% cancer growth inhibition due to 6b) ¨ (% cancer
growth inhibition due
to 6a)]. Pearson correlation coefficient, R = 0.3125, p = 0.017949,
significant at p <0.05.
There is a negative correlation between % cancer growth inhibition of 6a and
the product [(%
cancer growth inhibition due to 6b) ¨ (% cancer growth inhibition due to 6a)].
Pearson
correlation coefficient, R = -0.4211, p = 0.00111, significant at p <0.05.
Perhaps relevantly, around the chiral carbon, 6b and Econazole share 3 (of 4)
of the same
groups, wherein Econazole racemizes fast [361], faster than predicted by the
methodology of
[221].
Figure 31: Mechanistic distinction from oligomycin enables therapeutic
utility. Drawn
molecules of this figure (31A) have an imidazole group, with a protonable
nitrogen atom that
can shuttle protons across the mitochondrial inner membrane (IM), dissipating
the proton
motive force (pmf, uncoupling). This figure presents experimental data (31B)
using the HL-1
cardiac muscle cell line (cancer derived, but now very cardiac differentiated
e.g.
spontaneously contracts and beats like heart cells). Refer to the "Benchmark
Drugs" first,
which produce cellular effects well known to those of the art [3]. Oligomycin
here refers to
Oligomycin B. Oligomycin binds ATP synthase and blocks its forward, proton
passing, ATP
synthesizing, mode. This means less protons pass through ATP synthase, less
pmf consumed
per unit time, pmf increases, Tim hyperpolarizes, electron flow along the
respiratory chain
slows, and 02 consumption is decreased. Carbonilcyanide p-
triflouromethoxyphenylhydrazone (FCCP) is an uncoupler that shuttles protons
across the
IM, dissipates pmf (as heat), pmf decreases, Tim depolarizes, electron flow
along the
respiratory chain speeds, and 02 consumption is increased. Distinct from
oligomycin, three
molecules of this figure increase, rather than decrease, 02 consumption, which
signifies their
mechanistic distinction from oligomycin: they reduce ATP synthesis more by
uncoupling
than any inhibition of forward mode ATP synthase. They all contain a
protonable nitrogen
atom, with a basic pKa value conducive to uncoupling i.e. a pKa value
reasonably close to
{pH of mitochondrial intermembrane space + pH of mitochondrial matrix)/2).
Although for
VG025, its most conducive pKa is on its main ring rather than its imidazole.
In a NADPH-
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CA 3050553 2019-07-25
linked sub-mitochondrial (SMP) assay of ATP synthesis, these molecules would
decrease
ATP production because they dissipate pmf as heat, and so there is less pmf
available for
ATP production. In interpretation, this uncoupling could be incorrectly
attributed to
inhibition of the forward mode of ATP synthase and so the full mechanistic
distinction of
.. these molecules from oligomycin could be missed. This has been the case
with other
imidazole containing compounds of this disclosure. Also with a protonable
nitrogen in their
imidazole, also with a pKa conducive to uncoupling, and wherein their
inhibition of FIN
ATP synthesis in the NADPH-linked SMP assay has been attributed to inhibiting
the forward
mode of ATP synthase [5-8]. But wherein their uncoupling is likely to be the
more
predominant factor (extrapolated from data of this figure) and wherein they do
not inhibit the
forward mode of ATP synthase much, if at all, in stark distinction to
oligomycin. Uncoupling
capability, which decreases with increased logP (refer next paragraph),
explains why
different molecules of the present figure exert different effects on 02
consumption. The high
logP value of VG019 means its uncoupling is minimal and its effect on 02
consumption is
zero (rounded) at a concentration (100 M) it inhibits the reverse mode of ATP
synthase, in
stark distinction to oligomycin (3 i.iM), which dramatically decreases 02
consumption (-
40%), because, distinctly, it potently inhibits the forward mode of ATP
synthase.
LogP = ¨3.2 is the optimal compromise for best passing a membrane: its
hydrophobic core
(selecting for high logP) and hydrophilic boundary layer (selecting for low
logP) ([36], herein
incorporated in its entirety). The imidazole containing molecules presented in
this figure, and
in this disclosure's drawings more generally, have logP > 3.2 and present
increased logP =
decreased uncoupling. The uncoupling capability/liability of a molecule
actually hinges on its
intersection of pKa(s) and logP [36] but for the molecules in this
disclosure's drawings,
wherein the imidazole pKa values are, generally, all within a fairly narrow
range, the more
primary determinant to each molecule's uncoupling rate, relative to the
others, is the
molecule's logP value relative to the others.
The drawn molecules of this figure do inhibit the reverse mode of ATP
synthase. When a
respiratory chain inhibitor blocks electron flow, TIM is maintained, not by
proton pumping by
the respiratory complexes, but by proton pumping by ATP synthase i.e. the
reverse mode of
ATP synthase. In the presented data, when the respiratory chain is blocked,
the presented
molecules depolarise 'Pim because they block the reverse mode of ATP synthase.
They do not
affect Tim by these means when the respiratory chain is operational. Because
Tim is not
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CA 3050553 2019-07-25
set/maintained by the reverse mode of ATP synthase in this case. Although the
molecules
with stronger uncoupling capability, they can shuttle more protons across the
IM (dissipate
more pmf) than the respiratory chain can increase its rate to replace, and
they do depolarise
Tim. When the respiratory chain is blocked, a stronger uncoupler in this
figure depolarises
Tim more. Because not only does it inhibit the generator of Tim (reverse mode
ATP
synthase), it simultaneously erodes Tim (uncoupling).
Oligomycin does inhibit the reverse mode of ATP synthase. But distinctly it
inhibits its
forward mode more [11]. So, using oligomycin, there is no margin to inhibit
the reverse mode
(anti-cancer), without adversely affecting cells using OXPHOS i.e. most normal
cells.
Contrast this with molecule VG019 of this figure, for example, which can
inhibit the reverse
mode of ATP synthase, and yet ¨ in observed distinction to oligomycin ¨ does
not affect cells
using OXPHOS: it does not change their 02 consumption or Tim (at 100 M). This
grants it,
in distinction to oligomycin, anti-cancer selectivity. Other molecules of this
disclosure have
even greater cancer selectivity. For example, a preferred embodiment (refer
disclosure
section: "Preferred Embodiments") inhibits FIR) ATP hydrolysis >5,556 times
more than
FIFO ATP synthesis, in NADH-linked and NADPH-linked SMP assays [5-6], whilst
oligomycin ¨ inversely - inhibits F1F0 ATP hydrolysis less than FiFo ATP
synthesis in such
assays [11].
Computational calculations of logP and pKa were made using [25]. The data
presented in this
Figure is from [12] (herein incorporated in entirety), but the
analysis/(re)interpretation is
novel. As is the process/method of using these molecules as anti-cancer
therapeutics, which is
componentry to this invention. The imidazole of the drawn molecules is 4-yl.
Permutations,
with 5-y1 instead, are also disclosed by this invention as anti-cancer
therapeutics.
Figure 32. Spectra for the separated stereoisomers, Stereoisomer 1 and
Stereoisomer 2, of
structure 19a (shown in 32A, synthesised by Scheme 2 of this disclosure,
Expected/calculated
m/z {using "exact molecular weight" calculator in MarvinSketch software
[Chemaxon,
Budapest, Hungary, [25]], which uses the mass of the most prevalent isotope of
each element
in the molecule}: 537.08 [M+H], 559.06 [M+Na], 269.04 [M+2F1]2+, 575.03
[M+K]+). An
invention embodiment is the use of a compound/stereoisomer(s) with chiral SFC-
MS and/or
LC-MS and/or NMR feature(s) as presented in this figure for use in a method of
treatment of
the human or animal body by therapy, optionally for cancer
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CA 3050553 2019-07-25
treatment/amelioration/prevention/combat in a subject. (32B) Chiral
Supercritical Fluid
Chromotography Mass Spectrum (chiral SFC-MS) for Stereoisomer 1 sample. Upper
panel:
chiral-SFC annotated with Retention Time (RT, minutes) and area under peak.
Stereoisomer
1 is found in [RT=2.365] peak. Middle panel: component of mass spectrum of
[RT=2.365]
peak (polarity positive; ESCiTm is multi-mode ionization {PCT/US03/16892},
which
switches at very high speed between Electrospray Ionization (ESI) and
Atmospheric Pressure
Chemical Ionization (APCI) ionization methods, and ESCi+ refers to using ESCi
in its
positive ionization mode), wherein y-axis presents intensity and therein is
m/z 536.839
[M+H] (0.19 removed from Expected, better concordance in subsequent spectra).
Bottom
.. panel: component of mass spectrum of [RT=2.701] peak. Using this figure,
Stereoisomer 1
excess (%) in this sample = (80589248/(80589248+1727802))*100 = 97.9%;
Enantiomeric
excess (ee, %) = (97.9-50)*2 = 95.8%. Calculating the ee directly, rather than
via the
stereoisomer excess, ee = ((80589248-1727802)/(80589248+1727802))*100 = 95.8%.
Working back from this ee to the stereoisomer excess: (95.8/2)+50 = 97.9%.
(32C) Using this
figure, using UV Chromatogram component (more accurate), Stereoisomer 1 excess
(%) in
this sample = (19.93/(19.93+0.36))*100 = 98.23%; ee = (98.23-50)*2 = 96.46%.
(32D)
Liquid Chromatography¨Mass Spectrometry (LC-MS) spectrum for Stereoisomer 1.
Upper
panel: Liquid Chromatography (LC) Retention Time (RT) = 2.516 minutes. Lower
panel:
Mass Spectrum (MS) of [RT peak=2.516]. Shows positively charged species:
"Ionization
mode = API-ES" and "Polarity = Positive" wherein API is "Atmospheric Pressure
Ionization"
and ES is Electrospray. Percentages are relative abundance. (32E) MS
components. Upper
panel: m/z 537.1 [M+H]t 2nd panel: m/z 559.1 [M+Na]. 3rd panel: m/z 269.1
[M+2F1]2+.
Bottom panel: m/z 575.1 [M+K]. All these Observed m/z are within 0.07 of
Expected. (32F)
1H NMR {NMR probe temperature = 298.15 K} for Stereoisomer 1, scaled to
highest
compound peak, peaks identified by "Auto peak picking" algorithm (based on
Global
Spectral Deconvolution, GSD) of MestReNova version 12 software (Mestrelab
Research,
Santiago de Compostela, Spain), with default program settings used, NO
chemical structure
used/inputted to guide/constrain this peak picking. (32G) Aromatic region of
previous
spectrum, for Stereoisomer 1, expanded. (32H) chiral SFC-MS for Stereoisomer 2
sample.
Stereoisomer 2 is found in [RT=2.701] peak: m/z 536.839 [M+H] (0.19 removed
from
Expected). Stereoisomer 2 excess (%) = (435788880/(435788880+1578563))*100 =
99.64%;
ee = (99.64-50)*2 = 99.28%. (321) Using UV Chromatogram component (more
accurate),
Stereoisomer 2 excess (%) in this sample = (38.72/(38.72+0.46))*100 = 98.83%;
ee = (98.83-
50)*2 = 97.66%. (32J) LC-MS spectrum for Stereoisomer 2. RT = 2.516 minutes.
(32K) MS
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components. Upper panel: m/z 537.1 [M+H]t 2"d panel: m/z 559.1 [M+Na]t 3rd
panel: m/z
269.1 [M+2H]2+. Bottom panel: m/z 575.1 [M+K]t All these Observed m/z are
within 0.07 of
Expected. (32L) 1H NMR {NMR probe temperature = 298.15 K} for Stereoisomer 2.
"Auto
peak picking" algorithm in MestReNova v12 used to pick peaks shown. (32M)
Aromatic
region of previous spectrum, for Stereoisomer 2, expanded. (32N) Stereoisomer
1 in upper
panel, Stereoisomer 2 in lower panel. (320) Aromatic region, Stereoisomer 1 in
upper panel,
Stereoisomer 2 in lower panel.
Figure 33. Spectra for the separated stereoisomers, Stereoisomer A and
Stereoisomer B, of
Structure 8 in Scheme 7 of this disclosure (structure shown in 33A,
Expected/calculated m/z
{using "exact molecular weight" calculator in MarvinSketch}: 538.08 [M+Hr,
560.06
[M+Na], 269.54 [M+2H]2+, 576.04 [M+Kr). An invention embodiment is the use of
a
compound/stereoisomer(s) with chiral SFC and/or chiral SFC-MS and/or LC-MS
and/or
NMR feature(s) as presented in this figure for use in a method of treatment of
the human or
animal body by therapy, optionally for cancer
treatment/amelioration/prevention/combat in a
subject. (33B) Chiral Supercritical Fluid Chromotography Mass Spectrum (SFC-
MS) for
Stereoisomer A sample. Upper panel: chiral-SFC annotated with Retention Time
(RT,
minutes) and area under peak. Stereoisomer A is found in [RT=2.13] peak. Lower
panel:
component of mass spectrum of [RT=2.13] peak (ESCi+ is multi-mode ionization
{PCT/US03/16892} in positive ionization mode), wherein y-axis presents
intensity and
therein is m/z 538.215 [M+H] (0.135 removed from Expected, better concordance
in
subsequent spectra). (33C) Stereoisomer A excess = 100% (no other peak in the
UV
chromatogram), ee = (100-50)*2 = 100%. (33D) Liquid Chromatography¨Mass
Spectrometry
(LC-MS) spectrum for Stereoisomer A. Upper panel: Liquid Chromatography (LC)
Retention
Time (RT) = 2.685 minutes. Bottom panel: Mass Spectrum (MS) of [RT=2.685
peak], shows
positively charged species (API-ES positive), percentages are relative
abundance. (33E) MS
components. Upper panel: m/z 538.1 [M+H]t 2"d panel: m/z 560.1 [M+Na]+. 3rd
panel: m/z
269.6 [M+21-1]2 . 4th panel: m/z 576.1 [M+K]t All these Observed m/z are
within 0.06 of
Expected. (33F) High Resolution Mass Spectrometry (HRMS): Liquid
Chromatography-
Time of Flight (LC-TOF) Mass Spectrometry (MS) for Stereoisomer A. LC
Retention Time
(RT) = 0.166 minutes (LC data not shown). Upper panel: OBSERVED m/z
538.0745928061
[M+H], CALCULATED from structure m/z 538.0821456 [M+Hr, so Observed mass is
only
0.0075527939 Daltons (0.7% the mass of a proton) removed from calculation. The
species
with m/z 537.0671121449 [M+H] has hydrogen instead of deuterium on the chiral
carbon,
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and indeed is the mass of a neutron (1.008664 Daltons) lighter (538.0745928061-
537.0671121449) = 1.0074806612 Daltons mass difference (0.12% removed from
being the
exact mass of a neutron difference). Absolute abundance is shown on the y-axis
and the
molar percent deuterium incorporation for Stereoisomer A at the chiral carbon
is 100% -
((6180.43/710180.69)*100=0.87%) = 99.13%. 2nd panel: OBSERVED m/z
560.0600137508
[M+Na], CALCULATED from structure m/z 560.0640898 [M+Na], so Observed mass is
only 0.0040760492 Daltons (0.4% the mass of a proton) removed from
calculation. The
species with hydrogen instead of deuterium on chiral carbon is too low
abundance to be
observed here. Bottom panel: OBSERVED m/z 576.0250917093 [M+Kr, CALCULATED
from structure m/z 576.0380270 [M+K], so Observed mass is only 0.0129352907
Daltons
(1.3% the mass of a proton) removed from calculation. The species with
hydrogen instead of
deuterium on chiral carbon is too low abundance to be observed here. (33G) 11-
1 NMR {NMR
probe temperature = 301 K) for Stereoisomer A, scaled to highest compound
peak, peaks
identified by "Auto peak picking" algorithm of MestReNova v12 software, with
default
program settings used, NO chemical structure used/inputted to guide/constrain
this peak
picking. (33H) Aromatic region of previous spectrum, for Stereoisomer A,
expanded. (331)
The chiral SFC-MS machine broke down and so chiral SFC was performed in its
place for
Stereoisomer B, wherein RT = 4.535 min. (33J) LC-MS spectrum for Stereoisomer
B. RT =
2.685 minutes. (33K) MS spectrum components. Upper panel: m/z 538.1 [M+H]t 2'
panel:
m/z 560.1 [M+Na]+. 3"I panel: m/z 269.6 [M+2H]2+. Bottom panel: m/z 576.0
[M+K]. All
these Observed m/z are within 0.06 of Expected. (33L) HRMS: LC-TOF MS for
Stereoisomer B. LC Retention Time (RT) = 0.163 minutes (LC data not shown).
Upper panel:
m/z 538.0727757864 [M+H]t The species with m/z 537.0689210144 [M+H] has
hydrogen
instead of deuterium on the chiral carbon. Absolute abundance is shown on the
y-axis and the
molar percent deuterium incorporation for Stereoisomer B at the chiral carbon
is 100% -
((5782.74/669735.94)*100=0.86%) = 99.14%. 2nd panel: m/z 560.0513502753
[M+Na]t
Bottom panel: m/z 576.0327248583 [M+K]. (33M) 11-1 NMR {NMR probe temperature
=
300.7 K} for Stereoisomer B. "Auto peak picking" algorithm in MestReNova v12
used to
pick peaks shown. (33N) Aromatic region of previous spectrum, for Stereoisomer
B,
expanded. (330) Stereoisomer A in upper panel, Stereoisomer B in lower panel,
shows
integration values in this ppm region as automatically assigned by the
MestReNova v12
software used. Stereoisomer A has 2H assigned (1.03+0.93=1.96, rounds to 2),
Stereoisomer
B has 1H assigned (1.13, rounds to 1), in this ppm region. (33P) Shown here, I
manually
extended the integration line (N.B. this automatically changes the MestReNova
v12
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integration calculation method from "peak" to "sum") for Stereoisomer B to be
the same
length as that for Stereoisomer A, which changes its integration value from
1.13 (prior) to
1.52, which rounds to 2H. This improves the Stereoisomer B assignment because
now it has
the correct number of hydrogen nuclides (18H), whereas before it had
(incorrectly) 17H. It is
true that 1.52 narrowly crosses the threshold for 2H but interestingly, and
perhaps relevantly,
the integration value is also very low in this ppm region for the hydrogen on
chiral carbon
stereoisomers in Figure 32 (integration value at 7.93 ppm = 1.51 for
Stereoisomer 1, 1.57 for
Stereoisomer 2, as assigned by "Auto Assignment" algorithm of MestReNova v12).
(33Q)
'3C NMR {NMR probe temperature = 311.9 K} for Stereoisomer B. "Auto peak
picking"
algorithm in MestReNova v12 used to pick peaks shown. (33R) A region of
previous 13C
NMR spectrum, for Stereoisomer B, expanded. (33S) Stereoisomer A in upper
panel,
Stereoisomer B in lower panel. (33T) Aromatic region, Stereoisomer A in upper
panel,
Stereoisomer B in lower panel.
N.B. For Stereoisomer B, its LC-MS, HRMS and NMR are not completely aligned:
to
explain, LC-MS, HRMS and 'H NMR were performed and the NMR showed a high
amount
of impurities (spectrum not shown), further mass based purification steps were
taken, and the
1H NMR was repeated (spectrum shown herein), and a 13C NMR also, all showing a
much
lower, acceptable level of impurity, and it is this purer form that was
entered into anti-cancer
testing at the National Cancer Institute (NCI). However, LC-MS and HRMS were
not
repeated. But the LC-MS and HRMS are clear enough to identify the [M+ion]
species
needed, all be it with some additional species/impurities also observed that
aren't in the LC-
MS and HRMS spectra for Stereoisomer A (e.g. compare 33F and 33L).
Figure 34. Hydrogen vs. deuterium on chiral carbon. For 34A-34B: upper panel
shows 1I-1
NMR spectrum for Stereoisomer 1 of compound 19a synthesized by Scheme 2
(Figure 32),
lower panel shows 1H NMR spectrum for Stereoisomer B of Compound 8 synthesized
by
Scheme 7 (Figure 33). The former has a peak at 5.96 ppm from the hydrogen
attached to its
chiral carbon. The latter has this hydrogen replaced with a deuterium, and
thence this 5.96
ppm peak is absent in its 1H NMR spectrum.
Figure 35. Spectra for the separated stereoisomers, Stereoisomer a and
Stereoisomer p, of
Structure 8 in Scheme 13 of this disclosure (structure shown in 35A,
Expected/calculated m/z
{using "exact molecular weight" calculator in MarvinSketch}: 551.09 [M+Hr,
573.07
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[M+Na], 276.05 [M+2H]2 , 589.05 [M+K]-). Enantiomeric excess (ee) of both
stereoisomer
samples is >97%. An invention embodiment is the use of a
compound/stereoisomer(s) with
chiral SFC and/or chiral SFC-MS and/or LC-MS and/or NMR feature(s) as
presented in this
figure for use in a method of treatment of the human or animal body by
therapy, optionally
for cancer treatment/amelioration/prevention/combat in a subject. (35B) Chiral
SFC (AD-3
column, 1 ILL injected volume, co-solvent Me0H [0.05% IPAm]) for Stereoisomer
p sample,
RT = 1.9 min. (35C) Chiral Supercritical Fluid Chromotography Mass Spectrum
(SFC-MS)
for Stereoisomer a sample. Upper panel: chiral-SFC annotated with Retention
Time (RT,
minutes) and area under peak. Stereoisomer a is found in [RT=2.47] peak.
Middle panel:
component of mass spectrum of [RT=2.47] peak (ESCi+ is multi-mode ionization
{PCT/US03/16892} in positive ionization mode), wherein y-axis presents
intensity and
therein is m/z 551.290 [M+H] (0.2 removed from Expected, better concordance in
subsequent spectra). Bottom panel: component of mass spectrum of [RT=3.44]
peak.
Stereoisomer a excess (%) = (7100000/(7100000+7535))*100 = 99.89%; ee = (99.89-
50)*2
=99.78% (35D) Stereoisomer a excess = 100% (no other peak in the UV
chromatogram), ee
= (100-50)*2 = 100%. (35E) Liquid Chromatography¨Mass Spectrometry (LC-MS)
spectrum for Stereoisomer a. Upper panel: Liquid Chromatography (LC) Retention
Time
(RT) = 2.536 minutes. Bottom panel: Mass Spectrum (MS) of [RT=2.536 peak],
shows
positively charged species (API-ES positive), percentages are relative
abundance. (35F) MS
components. Upper panel: m/z 551.0 [M+H]t 2nd panel: m/z 573.0 [M+Na]. 3rd
panel: m/z
276.0 [M+2HF-. Bottom panel: m/z 588.9 [M+Kr. All these Observed m/z are
within 0.15 of
Expected. (35G) H NMR {NMR probe temperature = 298.2 K} for Stereoisomer a in
CDC13 solvent. "Auto peak picking" algorithm in MestReNova v12 used to pick
peaks
shown. The peak for the methyl group is likely to be around the peak for HDO
and so this
likely masks the methyl group from computational verification algorithm of
Mestrelab which
is why it doesn't concord that this spectrum corresponds with the structure
shown in 35A.
The large peak at 3.48 ppm corresponds to the ppm shift of methanol (Me0H) in
CDC13
solvent [362], wherein methanol was componentry to the mobile phase with the
chiral SFC,
and is likely here residual from this (labelling this as a known impurity
doesn't change the
negative finding of the Mestrelab verification algorithm). (35H) Aromatic
region of previous
spectrum, for Stereoisomer a, expanded. (351) IH NMR {NMR probe temperature =
298.2
ICI for Stereoisomer a in DMSO-d6 solvent. This spectrum was taken after HPLC
purification to try and remove impurities observed in the prior presented I H
NMR spectrum
for Stereoisomer a. "Auto peak picking" algorithm in MestReNova v12 used to
pick peaks
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CA 3050553 2019-07-25
shown. With DMSO-d6 solvent, the peak for the methyl group is not
masked/complicated by
a nearby solvent related (e.g. HDO) peak and the Mestrelab verification
algorithm does return
concordance between this spectrum and the structure shown in 35A. The large
peak at 2.08
ppm corresponds to the ppm shift of acetonitrile in DMSO-d6 solvent [362],
wherein the
mobile phase used for the HPLC consisted of acetonitrile and water, wherein
this peak is
likely residual from that process. (35J) Aromatic region of previous spectrum,
for
Stereoisomer a, expanded. (35K) Chiral SFC (AD-3 column, 3 IA injected volume,
co-
solvent Me0H [0.05% IPAm]) for Stereoisomer p sample, RT = 2.4 min. (35L)
Chiral
Supercritical Fluid Chromotography Mass Spectrum (SFC-MS) for Stereoisomer p
sample.
Upper panel: chiral-SFC annotated with Retention Time (RT, minutes) and area
under peak.
Stereoisomer 13 is found in [RT=3.44] peak. Middle panel: component of mass
spectrum of
[RT=3.44] peak (ESCi+ is multi-mode ionization {PCT/US03/16892) in positive
ionization
mode), wherein y-axis presents intensity and therein is m/z 551.290 [M+H] (0.2
removed
from Expected, better concordance in subsequent spectra). Bottom panel:
component of mass
spectrum of [RT=2.47] peak. Stereoisomer p excess (%) =
(4876384/(4876384+20876))*100
= 99.57%; ee = (99.57-50)*2 = 99.14%. (35M) Stereoisomer 13 excess = 100% (no
other peak
in the UV chromatogram), ee = (100-50)*2 = 100%. (35N) LC-MS spectrum for
Stereoisomer p. Upper panel: Retention Time (RT) = 2.540 minutes. Bottom
panel: Mass
Spectrum (MS) of [RT= 2.540 peak], shows positively charged species (API-ES
positive),
percentages are relative abundance. (350) MS components. Upper panel: m/z
551.1 [M+H].
2nd panel: m/z 573.0 [M+Na]+. 31d panel: m/z 276.1 [M+2H]2+. Bottom panel: m/z
588.9
[M+K]. All these Observed m/z are within 0.15 of Expected. (35P) 'H NMR {NMR
probe
temperature = 298.2 IC) for Stereoisomer 1 in CDC13 solvent. "Auto peak
picking" algorithm
in MestReNova v12 used to pick peaks shown. (35Q) Aromatic region of previous
spectrum,
for Stereoisomer 13, expanded.
Figure 36. Spectra for Compound 31, synthesized by Scheme Ha of this
disclosure (structure
shown in 36A, Expected/calculated m/z {using "exact molecular weight"
calculator in
MarvinSketch}: 555.14 [M+H], 577.12 [M+Na], 278.07 [M+2H]2+, 593.09 [M+K]+).
An
invention embodiment is the use of a compound with LC-MS and/or NMR and/or
HPLC
feature(s) as presented in this figure for use in a method of treatment of the
human or animal
body by therapy, optionally for cancer
treatment/amelioration/prevention/combat in a subject.
High Performance Liquid Chromatography (HPLC) Retention Time (RT) for Compound
31
= 2.117 minutes (HPLC spectrum not shown). (36B) Liquid Chromatography¨Mass
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Spectrometry (LC-MS) spectrum for Compound 31. Liquid Chromatography (LC)
Retention
Time (RT) = 0.87 minutes. Mass Spectrum (MS) of [RT=0.87 peak] shows
positively
charged species (API-ES positive), percentages are relative abundance. There
is an intense
peak at m/z 130.200 which might correspond to a N,N-Diisopropylethylamine
(DIPEA)
impurity ([DIPEA+H] mass is 130.16 Daltons using "exact molecular weight"
calculator in
MarvinSketch), wherein an impurity with a similar m/z value is observed in
other LC-MS
spectra herein for other compounds, thence this impurity might relate to the
machine and/or
LC-MS protocol used. (36C) MS components: Upper panel: m/z 554.90 [M+H]+. 2'
panel:
m/z 576.90 [M+Na]t Bottom panel: m/z 278.90 [M+2H]2 . No peak observed in
region of
[M+K] (not shown). First two m/z Observed are within 0.25 of Expected, last is
0.83
removed. (36D) 'H NMR (400 MHz, Methanol-d4) for Compound 31 {NMR probe
temperature = 297.9 IC}, scaled to highest compound peak, peaks identified by
"Auto peak
picking" algorithm of MestReNova v12 software, NO chemical structure
used/inputted to
guide/constrain this peak picking. (36E) Aromatic region of (36D) spectrum
expanded. (36F)
Downfield region of (36D) spectrum expanded. (36G) 1H NMR (400 MHz, Chloroform-
d)
for Compound 31 {NMR probe temperature = 298.5 K} . "Auto peak picking"
algorithm in
MestReNova v12 used to pick peaks shown. (36H) Aromatic region of (36G)
spectrum
expanded. (361)13C NMR (101 MHz, Methanol-d4) for Compound 31 {NMR probe
temperature = 298.0 K}. Firstly, "Auto peak picking" algorithm in MestReNova
v12 used to
pick peaks. Then, because the central Methanol-d4 solvent peak was observed at
49 instead
of expected 47.60 ppm, I manually assigned this 49 ppm Methanol-d4 solvent
peak as a
"reference peak" in the MestReNova v12 software, which shifted all ppm values
accordingly
(by [49-47.60] ppm), to become as presented in this spectrum. (36J) Upfield
region of (36I)
spectrum expanded. (36K) Downfield region of (36I) spectrum expanded. (36L) 1H-
13C
HSQC NMR (Me0D, 400 MHz, 101 MHz, F2 (proton dimension) acquisition
temperature =
299.8 K). (36M) Region of spectrum (36L) expanded. (36N) Region of spectrum
(36L)
expanded. (360)1H-13C HMBC NMR (Me0D, 400 MHz, 101 MI-k, F2 (proton dimension)
acquisition temperature = 298.9 K). (36P) Region of spectrum (360) expanded.
(36Q)
Region of spectrum (360) expanded. (36R) Assignments: atom numbers that follow
are
IUPAC numbers for the structure as it is shown in the figure: these
assignments come from
my analysis of the NMR data, especially utilising the 2D NMR spectra: 1H NMR
(400 MHz,
Methanol-d4) 5 (ppm) 8.35 (s, 1H, 35), 8.10 (m, 1H, 32), 8.04 (d, J = 2.1 Hz,
1H, 17), 7.78
(m, 2H, 16, 20), 7.27 (dt, J = 22.4, 7.4 Hz, 4H, 8, 10, 26, 28), 7.16 (dd, J =
8.0, 5.6 Hz, 2H,
11,27), 7.09 (d, J = 7.5 Hz, 2H, 25, 29), 7.03 (t, J = 7.4 Hz, 1H, 9), 4.71
(d, J = 13.9 Hz, 1H,
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4"), 4.37 (d, J = 14.2 Hz, 1H, 30'), 4.29 (d, J = 14.2 Hz, 1H, 30"), 3.96 (dd,
J = 19.7, 13.4 Hz,
2H, 2', 4'), 3.13 (m, 1H, 3), 2.78 (d, J = 12.8 Hz, 1H, 2"), 2.69 (ddd, J =
14.5, 9.7, 5.3 Hz, 1H,
23'), 2.54 (dt, J = 13.7, 8.6 Hz, 1H, 23"), 2.11 (s, 3H, 36), 1.50 (s, 1H,
22"), 1.36 (dtd, J =
14.1, 9.1, 5.1 Hz, 1H, 22').
13C NMR (101 MHz, Methanol-d4) (ppm) 149.14, 7, 142.91 , 24, 140.73, 13,
138.20, 19,
134.62 , 5, 134.51 ,20, 132.72 , 32, 132.22, 18, 130.62, 10, 130.05, 17,
129.75 , 8, 129.50,
26, 28, 129.39, 25, 29, 129.20, 31, 128.48 ,34, 127.87, 16, 127.01 , 27,
123.78 , 9, 121.75,
11, 58.88 , 3, 53.98 , 4, 53.69 , 2,48.11 ,30, 33.55 , 23, 31.58 , 22, 9.53 ,
36.
Figure 37: Spectra for compound synthesized by Scheme IIIa of this disclosure
(was
synthesized as an HC1salt, which dissociates in solvent, structure of most
prevalent (-100%)
free base tautomer is shown in 37A, Expected/calculated m/z {using "exact
molecular
weight" calculator in MarvinSketch}: 545.20 [M+H], 567.18 [M+Na], 273.10
[M+2H]2+,
583.15 [M+K]+). An invention embodiment is the use of a compound with LC-MS
and/or
NMR feature(s) as presented in this figure for use in a method of treatment of
the human or
animal body by therapy, optionally for cancer
treatment/amelioration/prevention/combat in a
subject. (37B) LC-MS spectrum. Retention Time (RT) = 1.763 minutes. Mass
Spectrum
(MS) of [RT=1.763 peak] shows positively charged species (API-ES positive),
percentages
are relative abundance. (37C) MS components: Upper panel: m/z 545.1 [M+H]. 2nd
panel:
m/z 567.1 [M+Na]+. 3rd panel: m/z 273.8 [M+2H]2+. Bottom panel: m/z 583.1
[M+K]t Three
of these Observed are within 0.1 of Expected, but the [M+2H]2+ species is 0.7
removed. Only
0.12% relative abundance is BMS-199264 without the methyl added (Observed m/z
530.90
[M+1-1]+, Expected m/z 531.18 [M+H]). (37D) 1H-13C HSQC NMR (CD3CN, 400 MHz,
101
MHz, F2 (proton dimension) acquisition temperature = 302.2 K). A potential
concern using
Scheme IHa is that the methyl group may attach to the 0 and not the imidazole
N atom of
BMS-199264. Well, in this 2D spectrum the [3.80 ppm (1H dimension) vs. 34.41
ppm (13C
dimension)] point is, with its 13C shift of 34.41 ppm, as apparent to someone
of the art, much
more likely to be methyl attached to the N, than the 0, atom. If the methyl
was attached to the
0 atom, the 13C shift would be closer to 50 ppm.
EXAMPLE EMBODIMENTS OF THE INVENTION
The Drawings present embodiments of the invention. Further examples are
enumerations of
Markush Formulas (I), (II), (III), (IV), (V) and (VI), presented henceforth.
Note: none of
these formulae share Markush symbols, which can be, for example, symbols of
the type: Rx,
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wherein x is an integer and/or letter. Such symbols are well recognised by
those of the art.
Markush Formulas (I), (II), (III), (IV), (V) and (VI), presented henceforth,
each have their
own Markush symbols, as specified for each, in their own respective sections
of this
disclosure. Further examples are protein embodiments of Formula (VII),
including
embodiments wherein one or more of a gene or nucleotide or DNA or RNA sequence
is
administered to the subject to administer a protein embodiment of Formula
(VII) to the
subject.
In this disclosure, the term "Formula [X]" is used when a statement is true
for Formula (I),
(II), (III), (IV), (V), (VI) and (VII), and all are being referred to
independently. A compound
of Formula [X] is a compound of Formula (I), or Formula (II), or Formula
(III), or Formula
(IV), or Formula (V), or Formula (VI), or Formula (VII), or any compound
presented in this
disclosure's Drawings, or any compound componentry to this invention.
This invention is described using these example embodiments but it isn't
limited to these.
These merely illustrate the invention. Compounds of other structures, which
are identified as
therapeutic inhibitors by the rationale and methods of the present invention,
are also
encompassed by the present invention.
An aspect of the invention is at least one compound of Formula (I), (II),
(III), (IV), (V), (VI)
or (VII) or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug
thereof, and/or a
pharmaceutical composition(s) comprising one or compounds of Formula (I),
(II), (III), (IV),
(V), (VI) or (VII), and/or an FIR ATP hydrolysis inhibitor(s) (that
optionally/preferably
inhibits FIFO ATP synthesis less or, more preferably, not at all), and/or a
compound(s)/composition(s) that reduces FiFo ATP hydrolysis, for use in
treating,
ameliorating, preventing, reversing or combating a disease or disorder, or
unwanted/undesirable physiological process or its consequences or an
unwanted/undesirable
aesthetic, selected from the following list;
Encompassed by this invention is a method of treating, ameliorating,
preventing, reversing or
.. combating a disease or disorder, or unwanted/undesirable physiological
process or its
consequences or an unwanted/undesirable aesthetic, in a subject, selected from
(i) cancer, any cancer, neoplasia, metastasis, tumor
formation/growth/implantation, tumorigenesis, solid tumor, blood borne tumor,
cancer that is
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refractory or resistant to conventional chemotherapy, drug resistant tumor,
multidrug resistant
cancer;
(ii) cancer that metabolizes much of its glucose and/or glutamine to lactate,
for
example a cancer exhibiting the Warburg effect and/or a cancer that can be
discriminated
from surrounding tissue by PET imaging (e.g. 18F-FDG PET);
(iii) cachexia, cancer driven cachexia, cancer fatigue, weight loss, weight
loss for
known or unknown reason, chronic wasting disease, atrophy, brown atrophy,
frailty, frailty
syndrome, aging frailty, geriatric syndrome, age-related cachexia and/or
sarcopenia,
weakness, wasting, anorexia nervosa, bulimia nervosa, eating disorder,
amenorrhea,
.. underweight, low body mass index (BMI, e.g. <18.5), low body fat
percentage, body
composition change, wasting syndrome, HIV wasting syndrome, malnutrition,
clinical
malnutrition, starvation, kwashiorkor syndrome, marasmus syndrome,
malabsorption,
malabsorption due to parasitic/bacterial infection (e.g. helminthiasis,
Whipple's disease, small
intestine bacterial overgrowth (SIB0), giardiasis etc.), anemia, refeeding
syndrome, appetite
loss, catabolysis, muscle atrophy, asthenia, muscle weakness (myasthenia),
sarcopenia,
osteoporosis, cachexia associated with HIV, AIDS, multiple sclerosis,
rheumatoid arthritis,
familial amyloid polyneuropathy, chronic kidney disease, cystic fibrosis,
multiple sclerosis,
motor neuron disease, Parkinson's disease, dementia, Addison's disease,
mercury poisoning
(acrodynia), chronic pancreatitis, untreated/severe type 1 diabetes mellitus,
hormonal
.. deficiency, tuberculosis, gastroenteritis, diarrhea, dysentery, any
digestive disease or
disorder, any gastrointestinal disease or disorder including functional
gastrointestinal
disorders, coeliac disease, tropical sprue, irritable bowel syndrome,
inflammatory bowel
disease, Crohn's disease, ulcerative colitis, short bowl syndrome, congestive
heart failure,
constrictive pericarditis, bradycardia, chronic obstructive pulmonary disease
(COPD),
.. altitude sickness, hyperthyroidism (subclinical hyperthyroidism, Graves'
disease,
multinodular goiter, toxic adenoma, inflammation of the thyroid {thyroiditis},
pituitary
adenoma), fatigue, chronic fatigue syndrome or any disease or disorder or
pathology in which
a body tissue(s) is undersupplied or underutilises (vs. its need) an
energetic/chemical
substrate(s), including 02;
(iv) cancer associated fever, which is especially associated with, but not
limited to,
non Hodgkin lymphoma (NHL), Hodgkin lymphoma, acute leukaemia, kidney cancer
(renal
cell cancer), liver cancer (hepatocellular carcinoma), bone cancer, adrenal
gland tumours
such as phaeochromocytomas, tumours in the hypothalamus, solid tumours;
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(v) disease or disorder or physiological process or condition that causes a
higher than
normal body temperature such as (without limitation) high environmental
temperature,
ingesting an uncoupler (e.g. 2,4-dinitrophenol), infection, sepsis,
neutropenic sepsis, stroke,
fever, pyrexia, hyperpyrexia, hyperthermia, malignant hyperthermia,
neuroleptic malignant
syndrome, serotonin syndrome, toxic serotonin syndrome, thyroid storm,
heatstroke, surgery
related, menopause ("hot flushes"), infection (non-limiting e.g. roseola,
measles, enteroviral
infections, parasitic, viral, fungal, Chlamydial, Rickettsial, bacterial,
mycobacterial, systemic
bacterial, intravascular, HIV associated, nosocomial), pyrogenic infection,
thermoregulatory
disorder(s), connective tissue disease(s), Kawasaki syndrome, drug overdose,
drug or drug
withdrawal induced hyperthermia, alcohol/drug withdrawal, idiosyncratic drug
reaction, fever
of known or unknown or uncertain origin (non-limiting e.g. infectious
disease(s),
inflammation, immunological disease(s), non-infectious inflammatory disease(s)
{non-
limiting eg. systemic rheumatic and autoimmune diseases, vasculitis,
granulomatous diseases,
autoinflammatory syndromes}, tissue destruction, reaction to incompatible
blood product(s),
metabolic disorder(s), inherited metabolic disorder(s), cancer, neoplasm,
endogenous or
exogenous pyrogen(s), injury, head injury);
(vi) disease/disorder/injury/pathology/surgery
treatable/ameliorated/prevented/combated/helped by conferring hypothermia in a
subject for
some medical or other purpose which can include slowing a chemical reaction(s)
rate in a
subject for therapeutic benefit, preventing/minimizing brain and/or tissue
damage, slowing
physiological/pathological processes (reaction rates are temperature
dependent) and so
"buying time" to get the subject to treatment for their emergency (e.g.
trauma/septic shock or
other medical emergency), slowing the progress of sepsis until a sufficient
concentration of a
working antibiotic(s) can be built up in the subject (furthermore hypothermia,
by slowing
sepsis progression, buys time to observe which antibiotic(s) can work,
yielding time to try
alternative further antibiotic option(s) if required), used soon after or just
before clinical/legal
death to preserve the subject's organs/tissues until the subject can be
frozen/cryogenically
frozen or the pathology that caused clinical/legal death (e.g. wound) can be
fixed and the
subject resuscitated, administered to a subject when a first responder (e.g.
ambulance crew)
deems the subject dead or unlikely to survive the journey to a medical
facility (e.g. hospital)
wherein this administration helps to preserve the subject which is helpful if
hospital staff
subsequently assess that they can, or might be able to, save the subject,
stabilizing
surgical/trauma/Emergency Room (ER) patients, deep hypothermic circulatory
arrest for
surgery (DHCA, non-limiting applications of DHCA include repairs of the aortic
arch, repairs
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CA 3050553 2019-07-25
to head and neck great vessels, repair of large cerebral aneurysms, repair of
cerebral
arteriovenous malformations, pulmonary thromboendarterectomy, resection of
tumors that
have invaded the vena cava, brain tumor resection {wherein the anti-cancer
activity of a
compound(s) of this invention juxtaposes well}), Emergency Preservation and
Resuscitation
(EPR), hypothermia for a surgical purpose, protective hypothermia during
surgery and/or
surgery complication, hypothermia to slow/reduce blood loss, hypothermia for
neuro- and/or
cardio- and/or organ/tissue and/or life protection in a subject that has
trauma/brain
trauma/polytrauma/surgery/stroke/ischemic stroke/hemorrhagic stroke/cardiac
arrest/myocardial infarction/hypoxia/shock (including, without limitation, low
volume,
cardiogenic, obstructive, and distributive shock)/sepsis/septic shock/multiple
organ
dysfunction syndrome/systemic inflammatory response syndrome (SIRS)/organ
failure/cytokine storm/anaphylactic shock/seizure/disseminated intravascular
coagulation/blocked
airway/croup/rhabdomyolysis/[head/facial/spinal/chest/abdominal/ballistic/knife
injury/trauma], or some other medical
emergency/condition/disorder/disease/injury/operation,
hypothermia for cardiac and/or cardiovascular surgery and/or open heart
surgery and/or brain
surgery (neurosurgery) and/or surgery using total circulatory arrest and/or
surgery using
cardiopulmonary bypass, Emergency Preservation and Resuscitation (EPR),
preserving
detached body parts such as limbs and/or organs (for example during organ
storage/transport
.. and/or transplant, thus increasing the time window for transplantation of
organs to recipients;
compound(s) of this invention is administered to organ to be transplanted [by
administration
to donor and/or by administration to isolated organ] and/or to organ
recipient, optionally
during transplant operation), protective hypothermia, targeted temperature
management,
therapeutic hypothermia, hypothermia therapy for neonatal encephalopathy,
neonatal
hypoxia-ischemia, birth asphyxia, hypoxic-ischemic encephalopathy (HIE),
haemorrhage,
hypovolemia, exsanguination, suspended animation, decompression sickness, burn
injury(s)
including skin burn, inflammation, allergic reaction, anaphylaxis,
tissue/organ rejection,
hypoxia, hypoxemia, anoxemia, anoxia, anemia, hypervolemia, altitude sickness,
obstructed
airway, asthma attack, hypoxia in a body/tissue/organ, hypoglycemia,
reperfusion injury
(ischemia-reperfusion injury), upon release of a ligature or tourniquet,
uraemia, crush
syndrome, compartment syndrome, traumatic brain and/or spinal cord injury,
major trauma,
infection, bacterial and/or viral infection(s) (non-limiting e.g. meningitis),
sepsis, septic
shock, stroke, cerebrovascular disease, ischemic brain injury, ischemic
stroke, traumatic
injury, brain injury, spinal cord injury, cardiac arrest, heart failure,
congestive heart failure,
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Dilated cardiomyopathy, valvular heart disease, pulmonary embolism, adrenal
crisis,
Addisonian crisis, hypertensive emergency, haemorrhagic (hypovolemic) shock,
cardiogenic
shock, neurogenic shock, hepatic encephalopathy, blood loss, ischemic
brain/heart/kidney/intestinal injury, neuroprotection and/or cardioprotection
and/or tissue
protection during/after a stroke and/or ischemia and/or cardiac arrest and/or
resuscitation
and/or a period(s) of poor blood flow anywhere in a subject;
(vii) poisoning by a toxic amount of a compound(s) in a subject (non-limiting
e.g.
carbon monoxide/methanol/heavy metal/ethylene glycol/pesticide poisoning,
snake/spider/bee/insect/lizard venom, metabolic poison(s), nerve agent,
chemical weapon,
bacterial toxin(s) (e.g. food poisoning, Salmonella poisoning), endotoxemia,
eukaryote
produced toxin(s) e.g. (non-limiting) brevetoxin, drug(s)/substance(s)
overdose e.g. (non-
limiting) heroin, ethanol, a prescription medication(s), an over the counter
medication(s) such
as aspirin, paracetamol etc.; hypothermia is protective to toxic insult);
(viii) hypermetabolism (optionally because of one or more of, without
restriction,
traumatic brain injury, injury to the body, infection, sepsis, burn, multiple
trauma, fever,
long-bone fracture, hyperthyroidism, prolonged steroid therapy, surgery, bone
marrow
transplant, recovery from anorexia/bulimia), heat intolerence, insomnia, fatal
insomnia,
nervousness, Luft's disease, non-thyroidal hypermetabolism, thyrotoxicosis,
hyperthyroidism,
overactive thyroid, subclinical hyperthyroidism, too much thyroid hormone(s)
in the subject,
too much triiodothyronine (T3) and/or thyroxine (14) in the subject,
hyperthyroxinemia
(including, without restriction, familial dysalbuminemic hyperthyroxinemia,
familial
euthyroid hyperthyroxinemia, thyroid hormone resistance syndrome), thyroid
storm,
hyperthyroidism caused by one or more of (without restriction) Graves'
disease, thyroiditis,
Hashimoto's thyroiditis, subacute thyroiditis, postpartum thyroiditis, lumps
(nodules) on the
.. thyroid, enlarged thyroid gland (goitre), simple goitre, multinodular
goiter, toxic multinodular
goiter, toxic adenoma, toxic thyroid adenoma, inflammation of the thyroid,
hyperplasia of
thyroid, metastatic thyroid cancer, thyroid tumour, thyroid cancer (including,
without
restriction, papillary carcinoma, follicular carcinoma, medullary thyroid
carcinoma,
anaplastic thyroid carcinoma), eating too much iodine, goitrogen ingestion,
consumption of
ground beef contaminated with thyroid tissue ("hamburger hyperthyroidism"),
too much
synthetic thyroid hormone in the subject, pituitary adenoma, drug induced,
Amiodarone drug
induced, struma ovarii, Jod-Basedow syndrome, nonautoimmune autosomal dominant
hyperthyroidism;
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(ix) low or less than desired metabolic/bioenergetic efficiency in a subject,
or low or
less than desired physical or mental performance (e.g. memory, IQ), or low or
less than
desired body weight, or fatigue/tiredness/weakness/exhaustion;
(x) accelerated aging disease or progeroid syndrome including, without
restriction,
Werner syndrome, Bloom syndrome, De Barsy syndrome, Rothmund¨Thomson syndrome,
Cockayne syndrome, xeroderma pigmentosum, trichothiodystrophy, combined
xeroderma
pigmentosum-Cockayne syndrome, restrictive dermopathy, Wiedemann¨Rautenstrauch
syndrome, Hutchinson¨Gilford progeria syndrome (progeria), Ataxia
telangiectasia-like
disorder 2, XFE progeroid syndrome, Muscular dystrophy, Muscular Dystrophy
(Becker's,
Duchenne, Limb-Girdle), Yamamoto's Muscular Dystrophy, Mandibuloacral
dysplasia,
Dilated cardiomyopathy, GAPO syndrome, Cutis laxia, Ehlers-Danlos syndrom,
Lenz-
Majewski hyperostatic dwarfism, SHORT syndrome, Progressive external
opthalmoplegia,
Nester-Guillermo progeria syndrome, MDPL syndrome, Dyskeratosis congenital,
Down
syndrome;
(xi) disease or disorder of aging (incidence/severity increases with increased
age/senescence) and/or unwanted/undesirable aspect(s) of aging and/or a
disease/disorder
associated with elevated reactive oxygen species including age-associated
decline, aging
frailty, frailty syndrome, sarcopenia, muscle weakness, osteoporosis,
cognitive decline,
cognitive defecit, mild cognitive impairment, degenerative diseases,
neurodegenerative
diseases, motor-associated neurodegenerative diseases, motor neuron disease,
amyotrophic
lateral sclerosis (ALS), primary lateral sclerosis, progressive muscular
atrophy, progressive
bulbar palsy, progressive supranuclear palsy, pseudobulbar palsy, hereditary
spastic
paraplegia, Parkinson's disease, Multiple System Atrophy (MSA), Progressive
Supranuclear
Palsy (PSP), essential tremor, resting tremor, Alzheimer's disease,
Huntington's disease,
spinocerebellar ataxias, Friedreich's ataxia, dementia, frontotemporal
dementia, chronic
traumatic encephalopathy, memory loss, aged cognition, age/aging related
cognitive
decline/impairment, Batten disease, polyglutamine diseases, osteoporosis,
atherosclerosis,
cardiovascular disease, myocardial infarction, cerebrovascular disease,
stroke, heart failure,
heart failure with preserved ejection fraction, idiopathic pulmonary fibrosis,
fibrotic disease,
pulmonary disease, coronary artery disease, hypercholesterolemia, obesity,
liver disease, fatty
liver disease, lysosomal storage disease, amyloidosis, systemic sclerosis,
kidney disease,
hepatic cirrhosis, immunosenscence, clonal hematopoiesis, chronic obstructive
pulmonary
disease (COPD), hypertension, hypercholesterolemia, age-related thymic
atrophy, arthritis,
osteoarthritis, arthritis (Osteo-and Rheumatoid), Juvenile Rheumatoid
Arthritis (JRA),
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Degenerative Disc Disease, Tendinopathy, Androgenetic Alopecia, male-pattern
baldness,
Idiopathic Pulmonary Fibrosis, systemic sclerosis, Chronic Obstructive
Pulmonary Disease,
Psoriasis, age-related loss of cardiac/pulmonary/cognitive/vision function,
diabetes, type 2
diabetes, andropause, glaucoma, retinal degeneration, sarcopenia, cachexia,
age-related
cachexia and/or sarcopenia, age-related macular degeneration (AMD,
early/intermediate/late), neovascular/wet AMD, dry AMD, Geographic atrophy
(GA), wet
and dry AMD in the same eye(s), Stargardt's macular degeneration, Best
vitelliform macular
dystrophy, diabetic retinopathy, proliferative diabetic retinopathy, diabetic
macular edema,
age/aging-related eye disease, ophthalmological disease/disorder, ocular
disease, vision loss,
progressive vision impairment, myopia (short-sightedness), degenerative
myopia, hyperopia
(far-sightedness), accommodative dysfunction, glaucoma, cataract formation,
retinal
degeneration, progressive retinal degeneration, retinitis pigmentosa, leber
hereditary optic
neuropathy, Fuchs spot, Best's disease, Sorsby's fundus dystrophy, hearing
loss (e.g. age-
related), presbycusis, tinnitus, naive T cell shortage, movement disability,
nonalcoholic fatty
liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), immunosenescence,
respiratory/urinary tract infection (RTI/UTI) especially in older/aged/elderly
subjects, cancer;
(xii) aging and/or one or more signs of aging, wherein one or more of these
compounds slow/delay/reduce/treat/prevent/stop/reverse aging, and/or extend
lifespan and/or
healthspan, and/or treat or delay the onset of geriatric aging of the
human/animal body,
tissue(s), or organ(s), and/or treat or delay the onset of an age-associated
phenotype in a
cell(s)/organism(s), and/or prolong fertility e.g. female fertility, delay
menopause;
(xiii) skin aging and/or damage (including sun damage) and/or scalp and/or
hair aging
and/or hair greying and/or hair loss;
(xiv) insomnia, fatal insomnia, sleep onset latency, delayed sleep phase
disorder,
exploding head syndrome, parasomnia, sleep-maintenance insomnia, sleep
disorder, too
much/inappropriate/undesired signals/activity/electrical activity in the
nervous system,
hyperactivity, hypersensitivity, Premature ejaculation, hyperreflexia,
Autonomic dysreflexia
(AD), Hyperventilation syndrome, brain hyperactivity, overly sensitive sensory
system,
pathological crying and/or laughing, Pseudobulbar affect (PBA, emotional
lability),
photophobia, phonophobia, temperature-sensitive, pressure-sensitive, brain
hyperexcitability,
overstimulation, intrusive thought(s), Perseveration, sensory overload,
disorganized thinking,
fantasy prone personality, malapdative daydreaming, dissociation, hyperkinetic
disorder,
agitation, Psychomotor agitation, restlessness, difficulty controlling
behaviour, disruptive
behaviour disorder, Emotional and behavioral disorder, pervasive developmental
disorder,
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Overactive disorder associated with mental retardation and stereotyped
movements, attention-
deficit disorder, Attention Deficit Hyperactivity Disorder (ADHD), adult
attention-deficit
hyperactivity disorder, severe behavioral problem(s) in children (e.g., to
illustrate and not
restrict, combativeness and/or explosive hyperexcitable behavior {out of
proportion to
.. immediate provocation[s]}, hyperactive children who show excessive motor
activity with
accompanying conduct disorders consisting of one or more of: impulsivity,
difficulty
sustaining attention, aggressivity, mood lability, poor frustration
tolerance), Premenstrual
dysphoric disorder (PMDD), premenstrual syndrome (PMS), impulsiveness,
impulsivity,
impulse control disorder, lack of self-control, hysteria, histrionic
personality disorder,
attention difficulty, inattention, poor attention control, anxiety, paranoid
anxiety, Paranoid
personality disorder, distress, dysphoria, Adjustment disorder, separation
anxiety, anxiety
disorder, depressive anxiety, agitated depression, treatment-resistant
depression, Generalized
anxiety disorder, social anxiety disorder, stranger anxiety, separation
anxiety (e.g. in dogs left
at home), separation anxiety disorder, Mixed anxiety-depressive disorder,
depression (all
forms, all severities), restlessness/apprehension/anxiety before surgery.
hypochondria, panic
disorder, panic attack, emotional outburst, emotional instability,
Intermittent explosive
disorder, unreasonable/unwarranted anger/aggression, hyper-aggression,
hostility, rage, poor
temper control, self-hatred, poor attentional control, worry, irritability,
neuroses, somatization
disorder, somatic symptom disorder, pain disorder, psychological pain,
psychogenic pain,
.. psychogenic facial pain, Atypical odontalgia (AO), burning mouth syndrome,
throbbing,
toothache/pulpitis/dental pain, chronic lower back pain, negative emotion,
persistent/enduring
negative emotion, body dysmorphic disorder, factitious disorder, illness
anxiety disorder,
unwarrented fight-or-flight response, stress, emotional stress, emotional
dysregulation,
distress, psychological stress, acute stress, chronic stress, acute stress
reaction, combat stress
reaction, traumatic grief, grief, grief after death of loved one, Prolonged
grief disorder (PGD),
heartbreak, guilt, shame, remorse, emotional pain, Algopsychalia, psychalgia,
suffering,
emotional trauma, psychological trauma, broken heart, Post Traumatic Stress
Disorder
(PTSD), Complex post-traumatic stress disorder (C-PTSD), hypervigilance,
sympathetic
hyperactivity, inability or impaired ability to relax, flashbacks, dysphoric
hyperarousal,
agoraphobia, insomnia, fatal insomnia, mood disorder, irrational/unwarrented
fear/terror,
phobia, social phobia, Cancerophobia, thunderstorm/firework phobia,
hypersexuality,
hypersexual disorder, depression, clinical depression, unipolar depression,
bipolar disorder,
Bipolar I, Bipolar II, Bipolar disorder not otherwise specified (Bipolar NOS),
cyclothymia,
cyclothymic disorder, mixed affective state, atypical depression, melancholic
depression,
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postpartum depression, double depression, seasonal affective disorder, mania,
manic episode,
hypomania, increase in energy of psychomotor activity, delirium, excited
delirium, major
depressive disorder, minor depressive disorder, recurrent brief depression,
Depressive
Disorder Not Otherwise Specified (DD-NOS), major depressive episode,
persistent
.. depressive disorder (PDD), dysthymia, dysthymic disorder, absence of
euthymia, manic
thoughts, racing thoughts, thought disorder, disordered thinking, reduced
ability to plan and
execute tasks, paranoia, hallucination (including, without limitation, visual,
auditory,
olfactory, gustatory, tactile, proprioceptive, equilibrioceptive, nociceptive,
thermoceptive,
chronoceptive), Charles Bonnet syndrome, delusion, persecutory delusion,
hearing voices,
.. homicidal/criminal ideation/tendency/thoughts, suicidal
ideation/tendancy/thoughts, self-
injury, non-suicidal self-injury, violence, attacking others, negative mood
swing, personality
disorder, Borderline personality disorder, Narcissistic personality disorder,
malignant
narcissism, dissociative disorder, dissociative identity disorder (DID),
Psychosis, acute
psychosis, chronic psychosis, psychosis spectrum disorder, manifestations of
psychotic
disorders, behavioral complications of mental retardation, stimulant
psychosis, psychotic
depression, hallucinogen persisting perception disorder, Psychoactive
substance-related
disorder, amphetamine-induced psychosis, brief psychotic disorder, Brief
reactive psychosis,
Menstrual psychosis, postpartum psychosis, Psychotic disorder, Psychopathy,
chronic
hallucinatory psychosis, manifestation(s) of psychotic disorder,
neurotic/reactive/endogenous/involutional/psychotic depression/depressive
disorder
(optionally accompanied by anxiety or agitation), depressive neurosis,
delusional depression,
psychotic aggression, psychiatric symptoms of dementia, AIDS delirium,
Supersensitivity
psychosis, Tardive psychosis, Tardive dysmentia, Depersonalization disorder,
out-of-body
experience, Sociopathy, Schizophrenia, Paranoid schizophrenia, disorganized-
type
.. schizophrenia, simple-type schizophrenia, pseudoneurotic schizophrenia,
prodromal
schizophrenia, schizoaffective disorder, bipolar type schizoaffective
disorder, depressive type
schizoaffective disorder, schizoaffective psychosis, Schizotypal personality
disorder,
schizophreniform disorder, Delusional parasitosis, formication, paresthesias,
Acroparesthesia,
tinnitus, delusional disorder, delusional jealousy, inappropriate behaviour,
behavioural
disorder, antisocial personality disorder, Oppositional defiant disorder
(ODD), conduct
disorder (CD), Disruptive mood dysregulation disorder (DMDD), sadistic
personality
disorder, poor inhibitory control, kleptomania, Pyromania, trichotillomania,
dermatillomania,
pathological/problem gambling, dyskinesia, tardive dyskinesia, paroxysmal
dyskinesia,
Paroxysmal kinesigenic dyskinesia, Paroxysmal nonkinesigenic dyskinesia,
Paroxysmal
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exercise-induced dystonia, Hemiballismus, tic disorder, tremor, clonus,
Tourette's syndrome,
coprolalia, copropraxia, Echophenomena, Echopraxia, Echolalia, Palilalia,
stuttering/stammering, stereotypy, punding, Self-stimulatory behaviour
(stimming),
Stereotypic movement disorder (SMD), synesthesia, obsession,
Obsessive¨compulsive
disorder (OCD), obsessive¨compulsive personality disorder, anankastic
personality disorder,
relationship obsessive¨compulsive disorder (ROCD), Scrupulosity, Primarily
obsessional
obsessive compulsive disorder, sexual obsession, Akathisia (including, without
limitation,
chronic, acute, Pseudoakathisia, Tardive akathisia, withdrawal or "rebound"
akathisia),
Restless legs syndrome, motor restlessness, periodic limb movement disorder
(PLMD),
periodic limb movements in sleep (PLMS), Sydenham's chorea, chorea, dystonia,
Hypnic
jerk, twitching, spasms, Tetanus, tetanus muscle spasms, tetanized state,
Myoclonus,
myoclonic seizure, Action myoclonus, Palatal myoclonus, Middle ear myoclonus,
Spinal
myoclonus, Stimulus-sensitive myoclonus, Sleep myoclonus, Cortical reflex
myoclonus,
Essential myoclonus, myoclonic epilepsy, Juvenile myoclonic epilepsy,
Progressive
myoclonus epilepsy (PME, including, without limitation, Dentatorubral-
pallidoluysian
atrophy, Unverricht-Lundborg disease, MERRF syndrome, Lafora disease),
Reticular reflex
myoclonus, Myoclonic epilepsy, diaphragmatic flutter, automatism, status
epilepticus,
Epilepsia partialis continua, Complex partial status epilepticus, epilepsy,
epileptic seizure,
simple partial seizure, complex partial seizure, generalized epilepsy,
generalized seizure
(including, without limitation, tonic-clonic, tonic, clonic, myoclonic,
absence (including
typical absence and atypical absence), atonic seizure), focal epilepsy, focal
seizure,
focal/partial seizure (including, without limitation, Simple partial seizure
and Complex partial
seizure), focal aware seizure, focal impaired awareness seizure, generalised
epilepsy,
temporal lobe epilepsy (including, without restriction, mesial temporal lobe
epilepsy
{MTLE} and lateral temporal lobe epilepsy ILTLEI), Frontal lobe epilepsy,
Rolandic
epilepsy, Nocturnal epilepsy, Nocturnal frontal lobe epilepsy, Autosomal
dominant nocturnal
frontal lobe epilepsy (ADNFLE), Generalized epilepsy with febrile seizures
plus (GEFS+),
Panayiotopoulos syndrome, epileptic fit, reflex epilepsy, reflex seizure,
absence seizure
(including, without limitation, childhood absence epilepsy, epilepsy with
myoclonic
absences, juvenile absence epilepsy, juvenile myoclonic epilepsy, Jeavons
syndrome {eyelid
myoclonia with absences}, genetic generalised epilepsy with phantom absences),
complex
partial seizure, atonic seizure, generalized tonic-clonic seizure,
tonic¨clonic seizure, extrinsic
stimulus epilepsy, intrinsic stimulus epilepsy, photosensitive epilepsy,
musicogenic epilepsy,
thinking epilepsy, eating epilepsy, seizure(s), Febrile seizure, nerve agent
induced seizure,
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Dravet syndrome (sometimes modest hyperthermic stressors like physical
exertion or a hot
bath can provoke seizures in affected individuals), acute symptomatic seizure,
seizure-related
disorder, drug related seizure, paroxysmal depolarizing shift, Ohtahara
syndrome, Epilepsy in
females with mental retardation, Rasmussen's encephalitis, Epilepsy syndrome,
benign
rolandic epilepsy, childhood absence epilepsy, absence epilepsy, juvenile
myoclonic
epilepsy, epileptic encephalopathies, Lennox¨Gastaut syndrome, West syndrome
(Epileptic
spasms), Doose syndrome (Myoclonic astatic epilepsy, MAE), Lennox-Gestaut
syndrome,
pseudo-Lennox Gastaut syndrome (atypical benign partial epilepsy), Benign
familial neonatal
epilepsy, Benign occipital epilepsy of childhood, familial neonatal
convulsions, Febrile
infection-related epilepsy syndrome, interictal dysphoric disorder, euphoria
sclerotic,
psychogenic non-epileptic seizure, non-epileptic seizure, gelastic seizure,
convulsion(s),
migraine, status migrainosus, tension headache, headache, Hypnic headache,
hiccups,
intractable hiccups, thumps in equines, Postural orthostatic tachycardia
syndrome (POTS),
Pheochromocytoma, agony, pain, chronic pain, acute pain, pain due to
disease/injury,
neuropathic pain, fibromyalgia, postherpetic neuralgia, phantom pain, referred
pain, back
pain, lower back pain, pelvic pain, cancer associated pain, chemotherapy
associated pain,
Diabetic neuropathy, small fiber peripheral neuropathy, Mononeuritis
multiplex,
Wartenberg's migratory sensory neuropathy, Breakthrough pain, idiopathic pain,
polyneuropathy, Neuritis, Mononeuropathy, Polyradiculoneuropathy, Radial
neuropathy,
neuropathy, visceral pain, Burning feet syndrome, Tarsal tunnel syndrome,
Carpal tunnel
syndrome, Guyon's canal syndrome, Cubital Tunnel Syndrome, Repetitive strain
injury,
Sciatica, Neurofibromatosis, Ulnar nerve entrapment, Erythromelalgia,
Paroxysmal extreme
pain disorder, Proctalgia fugax, dysesthesia, scalp dysesthesia, dysesthetic
burning,
hyperesthesia, hyperalgesia, Opioid-induced hyperalgesia, hyperpathia,
allodynia, pain
response from stimuli which do not normally provoke pain, Complex regional
pain syndrome
(said to be most painful condition known to man), Radiculopathy, neuralgia
(including,
without restriction, intercostal neuralgia, trigeminal neuralgia, atypical
trigeminal neuralgia,
glossopharyngeal neuralgia, occipital neuralgia, postherpetic neuralgia),
ciguatera poisoning,
irritable bowel syndrome (IBS), interstitial cystitis (IC), temporomandibular
joint disorder,
acute intermittent porphyria, Porphyria, Acute porphyria (including, without
limitation, acute
intermittent porphyria {AIP}, variegate porphyria {VP}, aminolevulinic acid
dehydratase
deficiency porphyria {ALAD}, hereditary coproporphyria {HCP}, drug induced),
Chronic
porphyria (including, without limitation, X-linked dominant protoporphyria
{XLDPP},
congenital erythropoietic porphyria {CEP}, porphyria cutanea tarda {PCT}, and
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CA 3050553 2019-07-25
erythropoietic protoporphyria {EPP}), cutaneous porphyria, Porphyria cutanea
tarda, allergy,
allergic reaction, anaphylaxis, anaphylactic shock, hives, asthma, allergic
rhinitis, rhinitis,
urticaria, contact dermatitis, cough, sore throat, esophageal reflux disease,
heartburn, chest
pain, Esophageal motility disorder, Nutcracker esophagus, diseases involving
gastrointestinal
motility, Peptic ulcer disease, esophageal spasm, angina, itchiness, Pruritus,
severe pruritus,
Prurigo, Pruritic skin condition, chronic itch, eczema, pruritus in eczema,
neuropathic itch,
neurogenic itch, itchiness due to atopic dermatitis and/or lichen simplex
chronicus, peripheral
sensitization, central sensitization, sensory perception of absent stimuli,
too much sensory
stimulation, sensory stimulation brings discomfort, Neuromyotonia, Peripheral
nerve
hyperexcitability, Morvan's syndrome, Benign fasciculation syndrome, Cramp
fasciculation
syndrome, hyperhidrosis, undifferentiated somatoform disorder, somatoform
disorder,
somatic symptom disorder, conversion disorder, functional neurological symptom
disorder,
severe nausea and/or vomiting, severe nausea/emesis, Chemotherapy-induced
peripheral
neuropathy, chemotherapy-induced nausea and vomiting (CINV), radiation therapy-
induced
nausea and vomiting (RINV), postnarcotic nausea, hyperemesis gravidarum,
morning
sickness, Cyclic vomiting syndrome, retching/dry heaving, Urinary
incontinence, enuresis,
nocturnal enuresis, tail chasing, reduce urine spraying/marking behavior,
benzodiazepine
withdrawal syndrome, barbituate withdrawal syndrome, Neuroleptic
discontinuation
syndrome, drug withdrawal discomfort/pain/symptoms, drug use disorder, alcohol
use
disorder, opoid use disorder, amphetamine use disorder, cocaine use disorder,
alcohol
withdrawal syndrome/symptoms, delirium tremens, opoid withdrawal
sydrome/symptoms,
drug craving, drug addiction, drug dependence, polysubstance dependence, drug
overdose,
smoking, tobacco (nicotine) addition, tobacco (nicotine) withdrawal symptoms,
alcoholism,
addiction, opoid addiction, cocaine/crack addiction, addictive behaviour,
addictive
personality, behavioural addiction, internet/computer/computer game/social
media/media
addiction, exercise addiction, compulsive behaviour (e.g. {non-limiting}
checking, counting,
washing, repeating), anti-social behaviour, criminality, sexual compulsion,
impulsive sexual
behaviour, compulsive buying, gambling addiction, sex related addiction,
sexual urge,
hunger, eating desire/compulsion, eating disorder, polyphagia, overeating,
binge eating
disorder, compulsive overeating, insatiable/excessive appetite, bulimia
nervosa, anorexia
nervosa, substance abuse, substance-induced delirium, substance-induced
psychosis,
substance-induced mood disorder, drug overdose, vertigo, motion sickness,
seasickness,
mentaUnervous breakdown, Autism spectrum disorder, neurological disorder,
cognitive
disorder, mental disorder, mental health disorder, mental health condition
involving impaired
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or altered neural plasticity, mood disorder, mental disorder disclosed in
Diagnostic and
Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) or a later
edition, a
mental/behavioural disorder disclosed by the International Classification of
Diseases (LCD) in
LCD-I 0 Chapter V: Mental and behavioural disorders (World Health
Organisation, WHO); or
(xv) diseases or disorders or conditions or pathologies or
unwanted/undesirable
effects/actions/behaviour treatable/ameliorated/prevented/combated, in
totality or in part (e.g.
along with surgery), by anaesthesia, pre-anaesthesia, post-anaesthesia,
hypoesthesia,
hypoactivity, sedation, coma, tranquilization, behavioural submission, muscle
relaxation,
hibernation, artificial hibernation, torpor, synthetic torpor, suspended
animation (e.g. used
during spaceflight because e.g. reduces ionizing radiation damage to subject);
(xvi) hyperproliferative/hyperplasia disorder, non-cancerous proliferative
disorder,
hyperproliferative autoimmune disorder, hyperplasia, epidermal hyperplasia,
dysplasia (e.g.
epithelial dysplasia), nodule(s), wart(s), papilloma(s), squamous cell
papilloma, genital
wart(s), condyloma(s), condyloma acuminatum, cyst(s), polyp(s) {including,
without
restriction, digestive, colorectal, endometrial, cervical, nasal, laryngeal,
inflammatory fibroid
polyp[s])}, inherited/hereditary (including, without restriction, Familial
adenomatous
polyposis, Peutz¨Jeghers syndrome, Turcot syndrome, Juvenile polyposis
syndrome, Cowden
disease, Bannayan¨Riley¨Ruvalcaba syndrome {Bannayan-Zonana syndrome},
Gardner's
syndrome) and non-inherited (non-restrictive e.g. Cronkhite¨Canada syndrome)
polyposis
syndrome, benign tumour, adenoma, organ enlargement by hyperplasia, Cushing's
disease
(enlarged adrenal cortex by hyperplasia), congenital adrenal hyperplasia,
hyperplasia of
breast, atypical ductal hyperplasia, intraductal papillomatosis,
fibroadenomas, fibrocystic
changes, hemihyperplasia, focal epithelial hyperplasia, sebaceous hyperplasia,
sebaceous
adenoma, intimal hyperplasia, unwanted/undesirable smooth muscle cell
proliferation,
smooth muscle cell hyperplasia, intimal smooth muscle cell hyperplasia,
neointimal
hyperplasia, proliferative vascular disorders, stenosis, stenosis because of
cellular
proliferation, vaginal stenosis, stenosis in a blood vessel, lessoned patency
of a blood vessel,
stenosis in a blood vessel because of cellular proliferation, vascular
occlusion, restenosis,
restenosis in a blood vessel that has been implanted with a stent, in-stent
restenosis, post-
angioplasty restenosis, systemic sclerosis, cirrhosis of the liver, adult
respiratory distress
syndrome, idiopathic cardiomyopathy, lupus erythematosus, retinopathy (e.g.
diabetic
retinopathy and/or other retinopathy[y/ies]), cardiac hyperplasia, fibrosis,
pulmonary fibrosis,
idiopathic pulmonary fibrosis, fibromatosis, neurofibromatosis, renal
interstitial fibrosis,
Cowden syndrome, hamartoma(s), choristoma(s), hemangioma(s), lymphangioma(s),
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CA 3050553 2019-07-25
rhabdomyoma(s), lymphangiomatosis, cystic hygroma, trichilemmoma, sarcoidosis,
neurosarcoidosis, aggressive fibromatosis, desmoid tumour(s),
unwanted/undesirable skin
cell proliferation, hyperproliferative skin disorder, psoriasis (including,
without restriction,
plaque, guttate, inverse, pustular, napkin, seborrheic-like, nail, scalp and
erythrodermic
psoriasis), psoriatic arthritis, dactylitis, seborrhoeic dermatitis, dandruff,
eczema, atopic
dermatitis, rosacea, reactive arthritis (Reiter's syndrome), pityriasis rubra
pilaris,
hyperproliferative variants of disorders of keratinization (e.g., without
restriction, actinic
keratosis, senile keratosis, keratosis pilaris, seborrheic keratosis),
scleroderma, benign
prostatic hyperplasia, prostate enlargement, endometrial hyperplasia, atypical
endometrial
hyperplasia, benign endometrial hyperplasia, adenomyosis, atypical polypoid
adenomyoma,
endometriosis, endometriosis of ovary (endometrioma), endometrial polyp(s),
polycystic
ovary syndrome, ovarian cyst(s), cervical polyp(s), uterine fibroid(s),
uterine hyperplasia;
(xvii) Tumour Associated Macrophages (TAMs) or any macrophage associated
disease or disorder such as, without limitation, Macrophage Activation
Syndrome (MAS),
HIV, AIDS, HIV-associated neurocognitive disorders (HAND), AIDS dementia, HIV
peripheral neuropathy, HIV associated cancers, AIDS-defining cancers, non-AIDS
defining
cancers, any disease in which the pathogen(s) hides from the immune system in
macrophages
including, without limitation, Mycobacterium tuberculosis (causes
tuberculosis), Leishmania
parasite (causes Leishmaniasis), Chikungunya virus (causes Chikungunya),
Legionella
pneumophila (causes Legionnaires' disease), adenoviruses, T. whipplei (causes
Whipple's
Disease), Brucella spp. (causes brucellosis), Staphylococcus aureus, Ebola
virus, Hepatitis B
virus, Hepatitis C virus, influenza virus strains, dengue virus and antibiotic
resistant bacteria,
any disease or condition in which activated macrophages are unwanted or
undesirable;
(xviii) virus/pathogen neuroinvasion via macrophage(s), as used for non-
limiting
example by HIV, Heptatitis C virus and SARS coronavirus;
(xix) neurocognitive or neurodegenerative diseases/disorders, for non-limiting
example those caused by a virus;
(xx) virus/pathogen transmission from mother to fetus/baby via macrophage(s)
as
used for non-limiting example by zika (via Hofbauer cells) and HIV
(macrophages in breast
.. milk);
(xxi) acute or chronic or systemic inflammation or any inflammatory
disease/disorder/syndrome or any autoinflammatory disease/disorder/syndrome or
any
autoimmune disease/disorder/syndrome;
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(xxii) acute inflammation, chronic inflammation, systemic inflammation,
inflammation because of infection or foreign bodies or injury or chemical or
toxin or drug or
stress or frostbite or burn or ionising radiation or surgery, inflammatory
diseases/disorders/syndromes, Macrophage Activation Syndrome (MAS),
autoinflammatory
.. diseases/disorders/syndromes, age-related chronic inflammatory diseases
("inflammaging"),
autoimmune diseases/disorders/syndromes, diseases/disorders of the innate
immune system,
sore throat, sore throat associated with cold or flu or fever, high-intensity
exercise associated
inflammation, ulcerative colitis, inflammatory bowel disease (IBD), irritable
bowel syndrome
(IBS), rheumatoid arthritis, osteoarthritis, inflammatory osteoarthritis,
psoriatic arthritis,
atopic dermatitis, allergic airway inflammation, asthma, inflammation
associated depression,
neuroinflammation, neuropathic pain, exercise-induced acute inflammation,
atherosclerosis,
allergy, hay fever, anaphylaxis, inflammatory myopathies, drug-induced
inflammation,
systemic inflammatory response syndrome, sepsis-related multiple organ
dysfunction/multiple organ failure, microbial infection, acute
brain/lung/hepatic/renal
injuries, lung inflammation, acute lung injury (ARDS), acne vulgaris, celiac
disease, celiac
sprue, chronic prostatitis, colitis, autoimmune hemolytic anemia,
diverticulitis,
glomerulonephritis, proliferative glomerulonephritis, membranous nephropathy,
minimal
change nephrotic syndrome, hidradenitis suppurativa, hypersensitivities,
interstitial cystitis,
Mast Cell Activation Syndrome, mastocytosis, otitis, pelvic inflammatory
disease (PID),
.. endometritis, reperfusion injury, rheumatic fever, rhinitis, sarcoidosis,
transplant rejection,
parasitosis, eosinophilia, type III hypersensitivity, ischaemia, chronic
peptic ulcer,
tuberculosis, Crohn's disease, hepatitis, chronic active hepatitis, immune
hepatitis, alcoholic
hepatitis, chronic viral hepatitis, ankylosing spondylitis, diverticulitis,
fibromyalgia, systemic
lupus erythematous (SLE), Alzheimer's disease, Parkinson's disease,
neurodegenerative
disease, cardiovascular disease, chronic obstructive pulmonary disease,
bronchitis, acute
bronchitis, bronchiectasis, bronchopneumonia, obliterative bronchiolitis,
appendicitis, acute
appendicitis, bursitis, cystitis, dermatitis, encephalitis, HIV encephalitis,
gingivitis,
meningitis, infective meningitis, myelitis, nephritis, neuritis,
periodontitis, chronic
periodontitis, phlebitis, prostatitis, RSD/CRPS, rhinitis, sinusitis, chronic
sinusitis, tendonitis,
.. testiculitis, tonsillitis, urethritis, vasculitis, respiratory
bronchiolitis¨associated interstitial
lung disease and desquamative interstitial pneumonia, pneumonia, interstitial
lung disease,
LOfgren syndrome, Heerfordt syndrome, monocytosis, liver fibrosis,
steatohepatitis,
nonalcoholic steatohepatitis, silicosis, histiocytoses, Langerhans' cell
histiocytosis,
haemophagocytic lymphohistiocytosis, pulmonary langerhans cell histiocytosis,
obesity, type
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II diabetes, gout, pseudogout, organ transplant rejection, epidermal
hyperplasia, chronic
fatigue syndrome, graft versus host disease (GVHD), lymphadenopathy,
rheumatoid arthritis
(RA), osteoarthritis (OA), inflammatory osteoarthritis, lupus, multiple
sclerosis (MS),
myocarditis, uveitis, CNS disease(s), inflammation aspect to a CNS disease(s),
hypothalamic
inflammation, dementia, glaucoma, amyloid related/driven disease, lipid
storage disease(s),
fibrosis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic
steatohepatitis (NASH),
cirrhosis, cirrhosis of the liver, alcoholic cirrhosis, nephropath(y/ies),
lupus nephritis,
immune nephritis, fibrotic disorder(s), cardiovascular disease, heart disease,
atherosclerosis,
vulnerable plaque, plaque formation, lipid containing macrophage related
disease(s)/disorder(s)/malad(y/ies), macrophage foam cells, diabetes, type 1
diabetes, type 2
diabetes, insulin resistance, macrophage aspect to insulin resistance,
obesity, obesity
associated inflammation, macrophage accumulation/large numbers of macrophages
in
adipose tissue (e.g. associated with obesity), granuloma(s), granulomatous
diseases,
sarcoidosis (including, without limitation, Annular sarcoidosis, Erythrodermic
sarcoidosis,
Ichthyosiform sarcoidosis, Hypopigmented sarcoidosis, LOfgren syndrome, Lupus
pernio,
Morpheaform sarcoidosis, Mucosal sarcoidosis, Neurosarcoidosis, Papular
sarcoid, Scar
sarcoid, Subcutaneous sarcoidosis, Systemic sarcoidosis, Ulcerative
sarcoidos),
neurosarcoidosis, pulmonary sarcoidosis, interstitial lung disease, pulmonary
fibrosis,
pulmonary tuberculosis, immune reconstitution syndrome of HIV,
Jarisch¨Hencheimer
reaction, sepsis, Paget's disease of bone, osteolysis, monocytosis,
histiocytosis, X-type
histiocytoses, non-X histiocytoses, Langerhans cell histiocytosis, non-
Langerhans-cell
histiocytosis, malignant histiocytosis, malignant histiocytic disorders,
histiocytomas,
histiocytic lymphoma, hemophagocytic syndrome, hemophagocytic
lymphohistiocytosis,
lymphohistiocytosis, diffuse histiocytic sarcoma, Rosai¨Dorfman disease,
gliosis, Bergmann
gliosis, Chronic Obstructive Pulmonary Disease (COPD), chronic inflammatory
lung disease,
familial mediterranean fever (FMF), TNF receptor-associated periodic syndrome
(TRAPS),
Hyperimmunoglobulinemia D with recurrent fever syndrome (HIDS), cryopyrin
associated
periodic syndrome (CAPS), Blau syndrome, Majeed syndrome, deficiency of
interleukin-1
receptor antagonist (D1RA), mevalonate kinase deficiency, pyogenic-arthritis-
pyoderma
gangrenosum and acne syndrome (PAPA), periodic fever aphthous stomatitis
pharyngitis
adenitis (PFAPA) syndrome, Behcet's disease, Still's disease, Crohn's disease,
Schnitzler's
syndrome, Sweet's syndrome, NLRP12-associated autoinflammatory disorders,
deficiency of
interleukin-1 receptor antagonist (DIRA), pyoderma gangrenosum, cystic acne,
aseptic
arthritis, periodic Fever Associated with mevalonate kinase deficiency
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(hyperimmunoglobulin D Syndrome), Pyogenic Arthritis Pyoderma Gangrenosum Acne
(PAPA) syndrome, Periodic Fever Aphthous Stomatitis, Pharyngitis and
Adenopathy
(PFAPA) syndrome, Adult-Onset Still's Disease (AOSD), Systemic Juvenile
Idiopathic
Arthritis (sJIA), Chronic Recurrent Multifocal Osteomyelitis (CRMO), Synovitis
Acne
Pustulosis Hyperostosis Osteitis (SAPHO) syndrome, Cryopyrin associated
Periodic
Syndrome (CAPS), Familial cold auto inflammatory syndrome (FCAS), Muckle-Wells
syndrome (MWS), Familial cold urticarial, Neonatal onset multisystemic
inflammatory
disorder (NOMID), hereditary Periodic Fever Syndromes, Periodic Fever
Syndromes,
systemic autoinflammatory diseases, Addison's disease, Agammaglobulinemia,
Alopecia
areata, Amyloidosis, Ankylosing spondylitis, Anti-GBM/Anti-TBM nephritis,
Antiphospholipid syndrome, autoimmune angioedema, autoimmune dysautonomia,
autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune inner ear
disease
(AIED), autoimmune myocarditis, autoimmune pancreatitis, autoimmune
retinopathy,
autoimmune urticaria, axonal & neuronal neuropathy (AMAN), BalO disease,
Behcet's
disease, benign mucosal pemphigoid, Bullous pemphigoid, Castleman disease
(CD), Celiac
disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy
(CIDP),
chronic recurrent multifocal osteomyelitis (CRMO), Churg-Strauss, Cicatricial
pemphigoid,
Cogan's syndrome, cold agglutinin disease, congenital heart block, coxsackie
myocarditis,
CREST syndrome, Berger's disease, dermatitis herpetiformis, dermatomyositis,
Devic's
disease (neuromyelitis optica), discoid lupus, Dressler's syndrome,
endometriosis,
eosinophilic esophagitis (EoE), eosinophilic fasciitis, erythema nodosum,
essential mixed
cryoglobulinemia, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant
cell arteritis
(temporal arteritis), giant cell myocarditis, glomerulonephritis,
proliferative
glomerulonephritis, membranous nephropathy, minimal change nephrotic syndrome,
Goodpasture's syndrome, granulomatosis with polyangiitis, Graves' disease,
Guillain-Barre
syndrome, Hashimoto's thyroiditis, hemolytic anemia, immune hemolytic anemia,
Henoch-
Schonlein purpura (HSP), Herpes gestationis or pemphigoid gestationis (PG),
hypogammalglobulinemia, IgA Nephropathy, IgG4-related sclerosing disease,
Immune
thrombocytopenic purpura (ITP), Inclusion body myositis (IBM), Interstitial
cystitis (IC),
juvenile arthritis, juvenile diabetes (Type 1 diabetes), juvenile myositis
(JM), Kawasaki
disease, Lambert-Eaton syndrome, Leukocytoclastic vasculitis, Lichen planus,
Lichen
sclerosus, Ligneous conjunctivitis, Linear IgA disease (LAD), Lupus, Lyme
disease chronic,
Meniere's disease, Microscopic polyangiitis (MPA), Mixed connective tissue
disease
(MCTD), Mooren's ulcer, Mucha-Habermann disease, Multiple sclerosis,
Myasthenia gravis,
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Myositis, Narcolepsy, Neuromyelitis optica, Neutropenia, Ocular cicatricial
pemphigoid,
Optic neuritis, Palindromic rheumatism (PR) PANDAS, Paraneoplastic cerebellar
degeneration (PCD), Paroxysmal nocturnal hemoglobinuria (PNH), Parry Romberg
syndrome, Pars planitis (peripheral uveitis), Parsonnage-Turner syndrome,
Pemphigus,
peripheral neuropathy, perivenous encephalomyelitis, Pernicious anemia (PA),
POEMS
syndrome, Polyarteritis nodosa, Polyglandular syndromes type I, II, III,
Polymyalgia
rheumatica, Polymyositis, Postmyocardial infarction syndrome,
Postpericardiotomy
syndrome, Primary biliary cirrhosis, Primary sclerosing cholangitis,
Progesterone dermatitis,
Psoriasis, Psoriatic arthritis, Pure red cell aplasia (PRCA), Pyoderma
gangrenosum,
Raynaud's phenomenon, Reactive arthritis, Reflex sympathetic dystrophy,
Relapsing
polychondritis, Restless legs syndrome (RLS), Retroperitoneal fibrosis,
Rheumatic fever,
Rheumatoid arthritis, Sarcoidosis, Schmidt syndrome, Scleritis, Scleroderma,
Sjogren's
syndrome, Sperm & testicular autoimmunity, Stiff person syndrome (SPS),
Subacute
bacterial endocarditis (SBE), Susac's syndrome, Sympathetic ophthalmia (SO),
Takayasu's
arteritis, Temporal arteritis/Giant cell arteritis, Thrombocytopenic purpura
(TTP), Tolosa-
Hunt syndrome (THS), Transverse myelitis, Type I diabetes, Ulcerative colitis
(UC),
Undifferentiated connective tissue disease (UCTD), Uveitis, Vasculitis,
Vitiligo, Wegener's
granulomatosis (or Granulomatosis with Polyangiitis (GPA)), idiopathic
thrombocytopenia
purpura, splenomegaly;
(xxiii) Systemic inflammatory response syndrome, cytokine release syndrome,
cytokine storm, immune reaction to a drug(s) or therapy(s), immune reaction to
an immune
activating drug(s) or agent(s) or treatment(s) or intervention(s), immune
reaction to
immunotherapy and/or immune-oncology and/or immunomodulatory drug(s) and/or
treatment(s), adverse reaction to adoptive T-cell therapy(s), adverse reaction
to a chimeric
antigen receptor T-cell therapy(s) (CAR-T cell therapy(s)), adverse reaction
to a immune
checkpoint inhibitor(s), adverse reaction to monoclonal antibody drug(s),
tumor lysis
syndrome;
(xxiv) cancer and Graft Versus Host Disease (GVHD) in transplantation therapy
in a
cancer patient;
(xxv) cardiovascular diseases and conditions associated with thrombosis and/or
the
formation of atherosclerotic plaques and/or ischemia and/or ischemic
conditions and/or
associated conditions including, without limitation, ischemia-reperfusion
injury, myocardial
ischemia, ischemic heart disease, chronic stable angina pectoris, first or
recurrent myocardial
infarction (MI), congestive heart failure, an acute coronary syndrome, muscle
cell damage,
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necrosis, cardiac arrhythmia(s), non-Q wave MI, unstable angina, high blood
pressure,
coronary artery disease, coronary arterial thrombosis, ischemic hypoxia,
cyanosis, gangrene,
acute limb ischemia, stroke, ischemic stroke, brain ischemia, vascular
dementia, ischemic
sudden death, transient ischemic attack (TIA), thrombophlebitis, ischemic
colitis, mesenteric
ischemia, angina pectoris, ischemic heart disease, ischemic neuropathy,
hypoxic-ischemic
encephalopathy, cerebral hypoxia, brain hypoxia, ischemia resulting from
vascular occlusion,
cerebral infarction, stroke and related cerebral vascular diseases (including
cerebrovascular
accident and transient ischemic attack), muscle cell damage, necrosis,
ventricular
hypertrophy, ventricular enlargement (including dilated cardiac myopathy and
heart failure),
Prinzmetal's angina, peripheral occlusive arterial disease (e.g., peripheral
arterial disease,
intermittent claudication, critical leg ischemia, prevention of amputation,
prevention of
cardiovascular morbidity such as MI, stroke or death), pericardial effusion,
constrictive
pericarditis, thrombosis, thrombotic or thromboembolic conditions, circulatory
disease
caused by blood clot (i.e. diseases involving fibrin formation, platelet
activation, and/or
platelet aggregation), thrombotic or thromboembolic symptoms of thromboembolic
stroke
(including that resulting from atrial fibrillation or ventricular mural
thrombus), arterial
cardiovascular thromboembolic disorders, venous cardiovascular thromboembolic
disorders,
thromboembolic disorders in the chambers of the heart, venous thrombosis
(including deep
vein thrombosis), arterial thrombosis, cerebral thrombosis, cerebral arterial
thrombosis,
pulmonary embolism, cerebral embolism, kidney embolism, arterial embolism,
thrombophilia, disseminated intravascular coagulation, restenosis, atrial
fibrillation,
atherosclerotic vascular disease, atherosclerotic plaque formation,
atherosclerosis,
atherosclerotic plaque rupture, peripheral arterial disease, coagulation
syndromes,
intermittent claudication, transplant atherosclerosis, vascular remodeling
atherosclerosis,
diabetic complications comprising retinopathy, nephropathy and neuropathy,
thromboembolic consequenses of surgery, interventional cardiology or
immobility,
thromboembolic consequenses of medication (such as oral contraceptives,
hormome
replacement and heparin), thrombotic consequenses of atherosclerotic vascular
disease and
atherosclerotic plaque rupture leading to tissue ischemia, prevention of
atherosclerotic plaque
formation, transplant atherosclerosis, thrombotic or thromboembolic
complications of surgery
including interventional cardiology, thromboembolic complications of pregancy
including
fetal loss, thromboembolic consequences of thrombophilia (e.g., Factor V
Leiden, and
homocystinenimia), prothrombotic consequences and/or complications of cancer,
prevention
of thrombosis on artificial surfaces (such as stents, blood oxygenators,
shunts, vascular access
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ports, vascular grafts, artificial valves, etc.), coagulopathies (e.g.,
disseminated intravascular
coagulation), coagulation syndromes, vascular remodeling atherosclerosis,
restenosis and
systemic infection, Kasabach-Merritt syndrome, occlusion (e.g. after a bypass)
and
reocclusion (e.g., during or after percutaneous transluminal coronary
angioplasty),
thromboembolic disorders resulting from conditions including but not limited
to
atherosclerosis, surgery or surgical complications, prolonged immobilization,
arterial
fibrillation, congenital thrombophilia, cancer, diabetes, effects of
medications or hormones,
complications of pregnancy and thrombosis resulting from prosthetic valves or
other
implants, indwelling catheters, stents, cardiopulmonary bypass, hemodialysis,
or other
procedures in which blood is exposed to an artificial surface that promotes
thrombosis; or
(xxvi) diseases or disorders or conditions
treatable/ameliorated/prevented/combated
by conferring/maintaining blood vessel patency in a subject, which can be
useful during
interventional cardiology or vascular surgery including bypass grafting,
arterial
reconstruction, atherectomy, vascular graft and stent patency, organ, tissue
and cell
implantation and transplantation, preservation of host and/or graft tissue as
related to organ
transplantation;
wherein the method comprises administering to the subject an effective amount
of at
least one compound of Formula (I), (II), (III), (IV), (V), (VI) or (VII) or a
pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof, and/or a pharmaceutical
composition(s)
comprising one or compounds of Formula (I), (II), (III), (IV), (V), (VI) or
(VII), and/or an
FiFo ATP hydrolysis inhibitor(s) (that optionally/preferably inhibits FIFO ATP
synthesis less
or, more preferably, not at all), and/or a compound(s)/composition(s) that
reduces FIE) ATP
hydrolysis.
The invention further includes the use of one or more of a
compound/composition disclosed
herein for the manufacture of a medicament, optionally for treating one or
more of the
diseases/disorders/conditions listed in the list immediately
above/aforementioned.
The present invention provides pharmaceutical compositions comprising a
pharmaceutically
acceptable carrier and at least one of the compounds of the present invention
or
stereoisomers, tautomers, pharmaceutically acceptable salts, or solvates
thereof. In an
embodiment, the present invention provides a composition comprising at least
one of the
compounds of the present invention or a stereoisomer, a tautomer, a
pharmaceutically
acceptable salt, or a solvate thereof. In another embodiment, the present
invention provides a
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pharmaceutical composition comprising a pharmaceutically acceptable carrier
and at least
one of the compounds of the present invention or a stereoisomer, a tautomer, a
pharmaceutically acceptable salt, or a solvate, thereof. In another
embodiment, the present
invention provides a pharmaceutical composition, comprising: a
pharmaceutically acceptable
.. carrier and a therapeutically effective amount of at least one of the
compounds of the present
invention or a stereoisomer, a tautomer, a pharmaceutically acceptable salt,
or a solvate
thereof. In another embodiment, the present invention provides a
pharmaceutical composition
further comprising additional therapeutic agent(s).
The compound(s)/composition(s) of the present invention can be used in
therapy. In another
embodiment, the present invention provides a compound of the present invention
or a
stereoisomer, a tautomer, a pharmaceutically acceptable salt, or a solvate
thereof, for use in
therapy. The compound(s)/composition(s) of the invention can be used in the
treatment
and/or prophylaxis of cancer. In another embodiment, the present invention
provides a
compound of the present invention or a stereoisomer, a tautomer, a
pharmaceutically
acceptable salt, or a solvate thereof, for use in therapy for the treatment
and/or prophylaxis of
cancer. In another embodiment, the present invention provides a method for the
treatment
and/or prophylaxis of cancer comprising administering to a patient in need of
such treatment
and/or prophylaxis a therapeutically effective amount of at least one of the
compounds of the
present invention or a stereoisomer, a tautomer, a pharmaceutically acceptable
salt, or a
solvate thereof. The compound(s)/composition(s) of the present invention can
be used for the
manufacture of a medicament, optionally a medicament for the treatment and/or
prophylaxis
of cancer. In another embodiment, the present invention provides the use of a
compound of
the present invention or a stereoisomer, a tautomer, a pharmaceutically
acceptable salt, or a
.. solvate thereof, for the manufacture of a medicament for the treatment
and/or prophylaxis of
cancer. The compounds/compositions of the invention can be used alone, in
combination with
other compounds/compositions of the present invention, or in combination with
one or more,
preferably one to two, other agent(s). In another embodiment, the present
invention provides
a combined preparation of a compound of the present invention and additional
therapeutic
agent(s) for simultaneous, separate or sequential use in therapy. In another
embodiment, the
present invention provides a combined pharmaceutical preparation of a compound
of the
present invention and additional therapeutic agent(s) for simultaneous,
separate or sequential
use in treatment and/or prophylaxis of cancer. In a preferred embodiment, the
additional
therapeutic agent(s) is an anti-cancer agent or combination thereof.
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CA 3050553 2019-07-25
In an embodiment, the present invention provides a process for making a
compound of the
present invention. In another embodiment, the present invention provides an
intermediate for
making a compound of the present invention. The present invention provides
processes and
.. intermediates for making the compounds of the present invention. All
processes used to
prepare compounds of the present invention and intermediates made therein are
considered to
be part of the present invention. The compounds of the present invention can
be prepared in a
number of ways known to one skilled in the art of organic synthesis. The
compounds of the
present invention can be synthesized using the methods described below,
together with
synthetic methods known in the art of synthetic organic chemistry, or by
variations thereof as
appreciated by those skilled in the art. Preferred methods include, but are
not limited to, those
described below. These schemes are illustrative and are not meant to limit the
possible
techniques one skilled in the art may use to prepare the compounds disclosed
herein.
Different methods to prepare the compounds of the present invention will be
evident to those
skilled in the art. Additionally, the various steps in the synthesis may be
performed in an
alternate sequence in order to give the desired compound or compounds. The
reactions are
performed in a solvent or solvent mixture appropriate to the reagents and
materials employed
and suitable for the transformations being effected. It will be understood by
those skilled in
the art of organic synthesis that the functionality present on the molecule
should be consistent
with the transformations proposed. This will sometimes require a judgment to
modify the
order of the synthetic steps or to select one particular process scheme over
another in order to
obtain a desired compound of the invention.
Example compounds are typically prepared as racemic mixtures. Preparation of
homochiral
.. examples may be carried out by techniques known to one skilled in the art.
For example,
homochiral compounds may be prepared by separation of racemic products by
chiral phase
preparative HPLC. Alternatively, the example compounds may be prepared by
methods
known to give enantiomerically enriched products. These include, but are not
limited to, the
incorporation of chiral auxiliary functionalities into racemic intermediates
which serve to
control the diastereoselectivity of transformations, providing enantio-
enriched products upon
cleavage of the chiral auxiliary.
It will also be recognized that another major consideration in the planning of
any synthetic
route in this field is the judicious choice of the protecting group(s) used
for protection of the
253
CA 3050553 2019-07-25
reactive functional group(s) present in the compounds described in this
invention. An
authoritative account describing the many alternatives to the trained
practitioner is Greene et
al. (Protective Groups in Organic Synthesis, 3rd Ed., Wiley-Interscience
(1999)). One of the
art is familiar with orthogonal protecting groups, orthogonality, wherein
different groups are
protected by different protecting groups, removable by different conditions,
and so a
protecting group(s) can be removed without removing another protecting
group(s).
EXAMPLE (I)
Summary of Formula (I)
This invention embodiment relates to compounds having the following formula:
Formula (I)
R2 R3
Z LM)rn X / (LN)1
'"=(LTh G2
(LU)u
w .õ,õG1 G3
ZZ (L c
(L.-)r ¨R1
OR
R2 R3
(LM)m (LN)n
Z N
(_1")t G2
(LU G1 ZN'' G3
)u
ZZ (LR)r ¨R1
OR
254
CA 3050553 2019-07-25
R2 R3
(LõN)m, ,,(LN)n
(LT)t G2*.
(LU)u4G3
I (LP)p
ZZ L (I-1r ¨R1
including
R2 R3
I
(LlmN (LN)n
(LT)t G2
(LU)u4 G3
I L
I
ZZ (Llr ¨R1
including
R2 R3
u (LN)n
zs=
(LlmI
(LTh NN I
(LU)u N G3
I
(L¨R1
=L H
(R4),4
including
R2N R3
N/
ONG3
L H
R1
(R4)q
255
CA 3050553 2019-07-25
or a pharmaceutically-acceptable salt, solvate, hydrate, or prodrug thereof,
wherein:
G1 is N or CH;
G4 is NH or Cl-I2;
G2 is N or CH;
G3 is sulphur (S) or oxygen (0) or selenium (Se) or CH2 and R1 is absent and r
is 0; or
G3 is nitrogen (N), or CH or phosphorus (P) and R1 is present;
example embodiments include
Z R2N.NrR3
L L
4110 N/%S
L H
(R4)q
,
Z R2NN"R3
L L
N X N
L H I
R1
(R4),4
;
Lm, LN, Lu, . T,
L LW, LP and LR are each independently selected from a single bond, 0, S, Se,
NRv, pRv, Be, c¨vs
(IC )2 or Si(Rv)2, wherein each Rv is independently selected from
hydrogen, deuterium, halogen (e.g. F), alkyl, or substituted alkyl (non-
limiting examples:
CF3, CC13), or deuterated alkyl (non-limiting example: CD3), or aminoalkyl, or
thioalkyl, or
alkoxy, or 0, or OH (hydroxyl), or halogen, or haloalkyl, or haloalkoxy;
m, n, u, t, w, p and r are each independently selected from 0, 1, 2, 3 and 4;
L is independently at each point of its use alkyl, or substituted alkyl (non-
limiting examples:
CF3, CC13), or deuterated alkyl (non-limiting example: CD3), or aminoalkyl, or
thioalkyl, or
alkoxy, or halogen, or haloalkyl, or haloalkoxy, or hydroxyalkyl or any atom
or isotope
permitted by valence (including any accompanying hydrogen(s)/deuterium(s) by
valence e.g.
(non-limiting) OH, NH2, SH, S1H3, PH2, BH2 etc.) including, without
limitation, La, Ti, Ce,
V, Ta, Cr, Mo, Mn, Fe, Ru, Os, Co, Pd, Pt, Cu, Ag, Au, Zn, B, Al, Ga, C, Si,
N, P, As, Sb, Bi,
0, S, Se, F, CI, Br, I, Hg;
256
CA 3050553 2019-07-25
R1 is absent, R.
- R3, hydrogen, deuterium, cyano, aryl, heteroaryl, ¨S02R8,
¨C(=0)R9,
¨C(=CH2)R9, ¨C(-0H)R9, ¨C(-SH)R9, ¨C(-SeH)R9, ¨C(-0L)R9 (wherein L defined
earlier), ¨C(=S)R9, ¨C(=Se)R9, ¨C(=NH)R9, C(=PH)R9, ¨S(=0)R9, ¨C(=N-OH)R9, ¨
C(-N=0)R9, ¨C(-P=0)R9, ¨C(=N-0-CH3)R9, --C(=RD)-(0-R9,
C(=RD)-R9, or
ic\z/R9
0 Se 0= s=
or or or or or
Se=
RD is 0, S, Se, NH or PH;
1.-1 is selected from a single bond, 0, S, NW or C(102, wherein each RJ is
independently
selected from hydrogen, deuterium, halogen (e.g. F), alkyl, or substituted
alkyl (non-limiting
examples: CF3, CCI3), or deuterated alkyl (non-limiting example: CD3), or
aminoalkyl, or
thioalkyl, or alkoxy, or halogen, or haloalkyl, or haloalkoxy;
j is 0, 1, 2 or 3;
Rextra is selected from L (defined earlier), aryl, heteroaryl, cycloalkyl,
heterocyclo, arylalkyl,
(heterocyclo)alkyl, (heteroaryl)alkyl, alkyl, substituted alkyl, alkenyl,
substituted alkenyl,
alkylene, substituted alkylene, alkynyl, substituted alkynyl, alkoxy,
thioalkyl, aminoalkyl,
carbamyl, sulfonyl, sulfonamide, cycloalkyl, (cycloalkyl)alkyl, hydroxyalkyl,
haloalkyl,
haloalkoxy, alkoxyalkyl, morpholinylalkyl, acyl, alkoxycarbonyl, substituted
amino;
R2 is (i) independently hydrogen, L (defined earlier), alkyl, or substituted
alkyl,
or (ii) taken together with R3 forms a heterocyclo;
R3 is (i) independently RI, alkyl, substituted alkyl, L (defined earlier),
alkylthio, aminoalkyl,
carbamyl, BB-aryl, BB-heterocyclo, BB-heteroaryl, or BB-cycloalkyl, or (ii)
taken together
with R2 forms a heterocyclo;
Z is heteroaryl;
Zz is aryl, heteroaryl, cycloalkyl or heterocyclo;
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CA 3050553 2019-07-25
BB is a bond, Ci_aalkylene, C2_4alkenylene, substituted Ci_aalkylene,
substituted C2-
4alkenylene, ¨C(=0)NR 9¨, ¨C _aalkylene-C(=0)NRI 9¨, or substituted C
1_4alkylene-
C(=-0)NR19¨;
114 at each occurrence is selected independently of each other R4 from the
group consisting of
PH2,0H, SH, halogen, alkyl, substituted alkyl, haloalkyl, nitro, cyano,
haloalkoxy, 0R25,
SR25, NR25R26, NR25S02R27, S02R27, S02NR25R26, CO2R26, C(=0)R26, CHNR25R26,
OC(=0)R25, ¨0C(=0)NR25R26, NR25C(=0)R26, NR25CO2R26, aryl, heteroaryl,
heterocyclo
and cycloalkyl;
R8 is alkyl, substituted alkyl, aryl, or heteroaryl;
R9 is ¨NR1ORII, alkyl, substituted alkyl, alkoxy, alkylthio, cycloalkyl, aryl,
heteroaryl,
heterocycle, heterocyclo or ¨CO2R12;
Rio and Rii, are (i) independently selected from hydrogen, alkyl, substituted
alkyl, alkoxy,
heterocyclo, cycloalkyl, aryl, and heteroaryl; or (ii) taken together form a
hetero
cyclo or heteroaryl;
Ri2 and R19 are hydrogen or alkyl;
R25 and R26 are independently selected from hydrogen, alkyl, or substituted
alkyl, or taken
together form a heterocyclo or heteroaryl ring;
R27 is alkyl or substituted alkyl, and
q is 0, I, 2, or 3.
Preferred compounds of Formula (I)
Preferred methods are to use, and preferred compounds are, compounds with the
following
formula, or pharmaceutically-acceptable salts, solvates, hydrates or prodrugs
thereof,
R2 R3
L
(R4),4 R1
further preferred methods are to use, and preferred compounds are, compounds
with the
following formula, or pharmaceutically-acceptable salts, solvates, hydrates or
prodrugs
thereof,
258
CA 3050553 2019-07-25
R3
R2 R3
oxµµH
S
N/kµN
R1
(R4)q
and even more preferred methods are to use, and preferred compounds are,
compounds with
the following formula, or pharmaceutically-acceptable salts, solvates,
hydrates or prodrugs
thereof,
R3
R2 R3
S _
N X N
R1
(R4)q
=
other preferred methods are to use, and preferred compounds are, compounds
with the
following formula, or pharmaceutically-acceptable salts, solvates, hydrates or
prodrugs
thereof,
R2'`=N/R3
41
N/IN.\
N CH3 R1
(R4)q H
further preferred methods are to use, and preferred compounds are, compounds
with the
following formula, or pharmaceutically-acceptable salts, solvates, hydrates or
prodrugs
thereof,
259
CA 3050553 2019-07-25
Z R2NN/R3
tik
\CH
s eix S
N N N
H'
R1
(R4 )q
other preferred methods are to use, and preferred compounds are, compounds
with the
following formula, or pharmaceutically-acceptable salts, solvates, hydrates or
prodrugs
thereof,
Z R2NN/R3
CH )
Sk x
eiiiN
li
H N N
I
(R4)q R1
other preferred methods are to use, and preferred compounds are, compounds
with the
following formula, or pharmaceutically-acceptable salts, solvates, hydrates or
prodrugs
thereof,
Z R2NN/R3
\,\F tik N
R N NN
HI
R1
(R4 )q
in which, in the preceding 7 structures shown:
L is hydrogen, or methyl, or alkyl, or hydroxyalkyl, or CF3, or CD3, or
deuterium (D);
D is deuterium (enrichment, for example, exceeding 40% deuterium incorporation
at shown
position, and optionally at other positions also);
S symbolises the S stereoisomer, for example, in enantiomeric excess (ee)
exceeding 70%;
R symbolises the R stereoisomer, for example, in enantiomeric excess (ee)
exceeding 70%
(following IUPAC naming rules the chiral carbon of the fluorine (F) analogue
is labelled R
rather than S, but note that the arrangement of which bond is up, bold wedge,
and down,
260
CA 3050553 2019-07-25
dashed, around the stereogenic carbon is the same as the preceeding structures
labelled S at
their chiral carbon, it is this molecule arrangement that is salient, and that
is disclosed, rather
than a mere label in a naming convention. This clarification won't be repeated
at every place
to which it applies in this disclosure, at every point at which there is an F
in place of an I-I on
the chiral carbon, or any other higher order of priority atom (by IUPAC
rules), e.g. (non-
limiting) any other halogen, because this clarification here itself is likely
superfluous: all this
is very clear to someone of the art. So, when there is said to be an
enantiomeric excess (ee) in
this disclosure in relation to this example embodiment, Formula (I), it
applies to this
molecular configuration, this arrangement of solid/dashed wedges, about the
chiral carbon,
whether this be S or R by IUPAC naming rules);
Z is triazolyl optionally substituted with one to two R7 or imidazolyl
optionally substituted
with one to two R7 and/or having fused thereto a benzene ring in turn
optionally substituted
with one to two R7;
Ri is cyano or ¨C(=0)R9;
R2 is hydrogen, alkyl, or benzyl;
R3 is aryl or arylalkyl optionally substituted with alkyl, halogen,
trifluoromethyl, OCF3,
cyano, nitro, amino, hydroxy, or methoxy;
R4 is halogen, alkyl, trifluoromethyl, or OCF3;
R7 is alkyl, carbamyl or carbamylCi_aalkyl;
R9 is ¨NRioRli, alkyl, substituted alkyl, alkoxy, alkylthio, cycloalkyl, aryl,
heteroaryl,
heterocycle, heterocyclo or ¨0O2R12;
Rio and Ri i are (i) independently selected from hydrogen, alkyl, substituted
alkyl, alkoxy,
heterocyclo, cycloalkyl, aryl, and heteroaryl; or (ii) taken together form a
heterocyclo or
heteroaryl;
R12 is hydrogen or alkyl; and
q is 0, 1, 2, or 3.
More preferred methods are to use, and preferred compounds are, compounds with
the
following formula, or pharmaceutically-acceptable salts, solvates, hydrates or
prodrugs
thereof,
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CA 3050553 2019-07-25
(R24)y
111110
Z R2 NV
(CHR23)x
N
*)k`
O (R4)q L N
R1
other preferred compounds are those with the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
(R24)y
Z NNV R2 (CHR23)x
Osoµ\H
lik S
N/kµN
I
(R4)q H R1
and even more preferred are compounds having the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
262
CA 3050553 2019-07-25
= (R24)y
Z R2NN," .õ(CHR23)x
*AD/ IN
jjk S N Ns= N
I
R1
(R4)q
other preferred compounds are those with the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
(11110 (R24)y
Z NNV R2 (CHR23)x
N -
/JO CH3 H I
(R4)q R1
other preferred compounds are those with the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
263
CA 3050553 2019-07-25
1110 (R24)y
Z R2N Z(CHR23)x
N
ACH;IN
.01
ak S ti,ii
N N
I
R1
(R4 )q
other preferred compounds are those with the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
11110 (R24)y
Z R2NNZ(CHR23)x
CH3
O R's1/4N//%N
I
(R4) (1H
R1
(1
other preferred compounds are those with the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
264
CA 3050553 2019-07-25
(R24)y
110
Z \NV R2 (CHR23)x
ox\F /IN
ik 12*
N X N
H
I
R1
(R4)q
in which, for the preceding 7 structures shown:
E
(R7a)0_3 I
Z is
%
%
Y¨N
and more preferably
,
265
CA 3050553 2019-07-25
R7c
R7a A syR7b R7a
Z iS or
Y ¨A ---- E A __
R7c
R7aNR7b R7a
or or
Y ¨N
+
R7a Nkr.R7b
or /7- or
Y¨N+ Y ¨N
L is hydrogen, or methyl, hydroxyalkyl, or CF3, or CD3, or deuterium (D);
D is deuterium (enrichment, for example, exceeding 40% deuterium incorporation
at shown
position, and optionally at other positions also);
S symbolises the S stereoisomer, for example, in enantiomeric excess (ee)
exceeding 70%;
A is nitrogen (N), or N+, or carbon;
E is absent, or alkyl, or substituted alkyl, or deuterated alkyl, or
aminoalkyl, or thioalkyl, or
alkoxy or any atom or isotope permitted by valence (including any accompanying
hydrogens
by valence e.g. (non-limiting) OH, NH2, SH, SiH3, PH2 etc.), for example
hydrogen,
deuterium or fluorine;
Y is N, CH or CR7c;
RI is cyano or ¨q=0)R9;
R2 is hydrogen or Cmalkyl;
R4 is halogen, Ci_zialkyl, trifluoromethyl; or OCF3;
R7a, R7b, and It7c are independently E (defined earlier), hydrogen, alkyl,
carbamyl or
carbamylCi_aalkyl, or R7a and R7c join to form an optionally substituted fused
phenyl ring;
R9 is ¨NR1ORII, alkyl, substituted alkyl, alkoxy, alkylthio, cycloalkyl, aryl,
heteroaryl,
heterocycle, heterocyclo or ¨0O21Z12;
266
CA 3050553 2019-07-25
Rio and Rii are (i) independently selected from hydrogen, alkyl, substituted
alkyl, alkoxy,
heterocyclo, cycloalkyl, aryl, and heteroaryl; or (ii) taken together form a
heterocyclo or
heteroaryl;
R12 is hydrogen or alkyl;
R23 is hydrogen, alkyl, hydroxyalkyl, or phenyl;
R24 is alkyl, substituted alkyl, haloalkyl, halogen, trifluoromethyl, cyano,
hydroxy, OCF3,
methoxy, phenyloxy, benzyloxy, cyano, or acyl, or two R24 groups join to form
a fused
cycloalkyl or benzene ring;
el is 1 or 2;
x is 0, 1, or 2; and
y is 0, 1, 2, or 3.
More preferred methods are to use, and preferred compounds are, compounds with
the
following formula, or pharmaceutically-acceptable salts, solvates, hydrates or
prodrugs
thereof,
IP (R24)y
Z z(CHR23)x
HN
ilk N N N
L H I
R1
(R4),1
other preferred compounds are those with the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
267
CA 3050553 2019-07-25
IP (R24)y
Z "(CHR23)x
HN
.0 \\ H
110 S N/N
R1
(R4 )q
and even more preferred are compounds having the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
e(R24)y
Z "(CHR23)x
HN
AD
O Ss NZN
I
R1
(R4)q H
other preferred compounds are those with the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
268
CA 3050553 2019-07-25
(R24)y
1110
Z y(CHR23)x
HN
SIPN/NN N
CH3 " I
(R4)q R1
other preferred compounds are those with the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
(R24)y
=
Z z(CHR23)x
HN
ACH3
O S' N/\\\N
I
(R4)4 H R1
other preferred compounds are those with the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
269
CA 3050553 2019-07-25
(R24)y
11110
Z y(CHR23)x
HN
IcaCH3",,IN
eft/ X
N N
H'
R1
(R4 )q
other preferred compounds are those with the following formula, or
pharmaceutically-
acceptable salts, solvates, hydrates or prodrugs thereof,
(R24)y
110
Z z(CHR23)x
HN
F\N
l io .0
H R1
R N/N
I
(R4)q
in which, for the preceding 7 structures shown:
270
CA 3050553 2019-07-25
R7c
R7a N=y.. R7b R7a
Z iS or
R7c
R7a N + R7b R7a
or
'5Y or \
N+
R7c R7c
L is hydrogen, or methyl, or hydroxyalkyl, or deuterium;
D is deuterium (enrichment, for example, exceeding 40% deuterium incorporation
at shown
position, and optionally at other positions also);
S symbolises the S stereoisomer, for example, in enantiomeric excess (ee)
exceeding 70%;
RI is cyano or ¨C(=0)R9;
R4 is halogen, Ci_aalkyl, trifluoromethyl, or OCF3;
R7c is hydrogen or R7 and R7c join to form a fused benzene ring optionally
substituted with
C _4a1ky1 or ¨(CH2)1.2¨NHC(=0)Ci_4a1ky1,
R7b is hydrogen, C1_4alkyl, or ¨(CH2)i_2¨NHC(=0)Ci_4alkyl;
R9 is a) ¨NRIoRi
b) Ci_salkyl optionally substituted with one to two of:
i) SR13, OR13, NRi3aRi3b, halogen, trifluoromethyl, CO2Ri3a, and
C(=0)NRi3aR13b,
ii) cycloalkyl optionally substituted with one to two of C(=0)H,
alkenyl, carbamyl,
and/or phenyl in turn optionally substituted with halogen;
iii) phenyl or napthyl optionally substituted with one to two of halogen,
nitro, amino, alkyl,
hydroxy, Ci_aalkoxy, or having fused thereto a five or six membered
heterocyclo;
iv) pyridinyl, thiophenyl, furanyl, tetrahydrofuranyl, or azepinyl, optionally
substituted with
alkyl or having fused thereto a five to six membered carbocyclic ring
optionally substituted
with keto or Ci_aalkoxy;
c) C i_aalkoxy;
d) Ci_4alkylthio;
e) CO2alkyl;
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CA 3050553 2019-07-25
f) 3 to 6 membered cycloalkyl optionally having up to four substituents
selected from alkyl,
halogen, cyano, alkenyl, acyl, alkylthio, carbamyl, and/or phenyl in turn
optionally
substituted with halogen; or having an aryl fused thereto;
g) phenyl optionally substituted with one to four of halogen, cyano,
trifluoromethyl, nitro,
.. hydroxy, Cmalkoxy, haloalkoxy, Ci_6alkyl, CO2alkyl, SO2alkyl, SO2NH2,
amino, NH(Ci-
4alkyl), N(Cmalky1)2, NHC(=0)alkyl, C(=0)alkyl, and/or C14 alkyl in turn
optionally
substituted with one to three of trifluoromethyl; hydroxy, cyano, phenyl,
pyridinyl; and/or a
five or six membered heteroaryl or heterocyle in turn optionally substituted
with keto or
having a benzene ring fused thereto;
.. h) pyridinyl, thiazolyl, furanyl, thiophenyl, and pyrrolyl optionally
substituted with one to
two of halogen, alkyl, and phenyl in turn optionally substituted with halogen
or
trifluoromethyl;
Rio is hydrogen, alkyl, or alkoxy;
Rii is alkyl, substituted alkyl, alkoxy, heterocyclo, cycloalkyl, aryl, or
heteroaryl;
or Rio and Rii, taken together form a heterocyclo or heteroaryl;
R23 is hydrogen, alkyl, hydroxyalkyl, or phenyl;
R24 is alkyl, halogen, trifluoromethyl, cyano, halogen, hydroxy, OCF3,
methoxy, phenyloxy,
benzyloxy, cyano, or acyl, or two R24 groups join to form a fused cycloalkyl
or benzene ring;
q is 0, 1, or 2;
xis 0 or 1; and
y is 0, 1, or 2.
Most preferred are compounds as immediately defined above wherein, Ri is cyano
or
-C(=0)R9; R9 is optionally substituted phenyl or phenyl Cmalkyl; x is 0 or 1;
and q and y
are 1 or 2. For this preferred structure, its S stereoisomer is preferred. And
further preferred is
for its L group to be deuterium.
Example embodiments of Formula (I)
Compounds from [5-6], selected as specific anti-cancer therapeutics by the
invention of this
disclosure, selected because they inhibit the reverse, more than the forward,
mode of ATP
synthase. EC50 and ICso used interchangeably. ECso values for FiFo ATP
hydrolysis, and FiFo
ATP synthesis, in NADH-linked and NADPH-linked sub-mitochondrial (SMP) assays
respectively, sourced from [5-6], are presented. [5-6] refer to these ECso
values as IC50 values
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CA 3050553 2019-07-25
for inhibiting FIFO ATP hydrolase (reverse mode) and Fi Fo ATP synthase
(forward mode).
However, this in incorrect. Because, as identified by the invention of this
disclosure,
explained herein, although these molecules inhibit FIFO ATP hydrolase, their
reducing of FiFo
ATP synthesis is not (predominantly) because of inhibiting FM ATP synthase,
but by
uncoupling. More preferred molecules of this invention have a low ECK' for
FiFo ATP
hydrolysis, and a higher ECK' for FiFo ATP synthesis, and their ratio
difference is large.
N
t_ I
0
CI
N NH
CI
SOD
Racemate CI
EC so FIFO ATP hydrolase = 0.033 0.02 (pM)
EC so RIF ATP synthesis > 100 (pM)
EC50 Ratio >3,030
N N
I
0 0
CI , HA CI .0õH
NH S N NH
CI
CI
4111
R stereoisomer Cl S stereoisomer Preferred CI
EC50FiFo ATP hydrolase > 100 (pM) EC so FiFo ATP hydrolase = 0.018 0.016
(pM)
EC50 FiFo ATP synthesis > 100 (pM) ECso F1F0 ATP synthesis > 100 (pM)
ECso Ratio >5,556
In rat: orally bioavailable (47%), iv. half-life (2.1 hours),
C = 21 pM, volume of distribution Vss = 2.37
Ukg
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CA 3050553 2 01 9-0 7-25
Further example embodiments of Formula (I), with SMP data, reinterpreted (as
aforementioned, these molecules don't significantly inhibit FIR) ATP synthase
but do reduce
FIFO ATP synthesis by uncoupling), from [5],
CI
N 0
CI CI
CI N//11\ NH
N).\ NH
ci 1.1 CI 41
CI CI
Ec50 Fi Fo ATP hydrolase = 0.082 t 0.03 (pM) EC50 FiF0 ATP hydrolase = 2.41
(pM)
EC50 F1F0 ATP synthesis > 100 (pM) EC50 F1F0 ATP synthesis > 100 (pM)
EC Ratio >1,220 EC Ratio >41.5
F N
FJJJci
N NH N NH
CI 41 CI
CI
41i
CI CI
EC FiFo ATP hydrolase = 0.71 t 0.34 (pM) EC F1F0 ATP hydrolase = 0.60 t 0.16
(pM)
EC F1 F0 ATP synthesis > 100 (pM) EC50 FiFo ATP synthesis > 100 (pM)
EC50 Ratio >141 EC Ratio >167
Further examples [5]:
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CA 3050553 2019-07-25
For all: EC50 F1F0 ATP synthesis > 100 OA
R1 R2 EC50F0 ATP hydrolase ( M)
4-CI CN 8.8
2-CI CN 2.23 /R2
2,3-Cl2 CN 2.49 0.72
3-CL CN 9.17
4-CI C(=0)4-CN-Ph 0.28 CI
4-CI C(=0)Et 2.27 NH
Ri
= H
CI
Further example, with synthesis step, effectively without a protonable element
in its
imidazole, which diminishes the molecule's ability to uncouple the proton
motive force:
N INCH3 N 0
N NH N NH
CI CI is 401
CI
Further example embodiment:
/NH
ci
N NH
CI
Further example embodiments:
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CA 3050553 2019-07-25
0
N11', N1/'''
\µ--N CI CICI .--N
0 N 0
CI I I
N NH NH
H
1410 0
CI CI
,
GENERAL COMPOUND SYNTHESIS
A general synthetic route applicable to some compounds of the invention is set
out in Scheme
/ below.
Scheme 1
Z
Z (a) Z-,, (b) L
3,H20
N H2
-,-'' 0 H DMF =-...4-----.'"----"--"-1N3
THE
1 (R4)q X'', (R4)4
SCN-R3
(R4 )(1
(C) CH3CN
Z R3
Z R,,, 3 KO Hil""'
H NI" L
H ACN, NH3,
H20 (R4)q
(R4)q I Rg-COOH
(e) CDI, THF
Z, ,R3
--- HN" Rg Z
HN3 R9
L
(n Lõ,,
I H
NNO
SEC s'µ
H
(R44' 0
(R4),:i
The person skilled in the art is able to make modifications to this general
synthetic route,
based on the common general knowledge, the chemical reaction literature,
and/or the content
276
CA 3050553 2019-07-25
of prior art disclosures cited herein, in order to synthesise compounds of the
invention where
necessary.
SPECIFIC COMPOUND SYNTHESIS
Racemate 19a [5] was synthesised by the following synthesis route, Scheme 2,
and separated
into component stereoisomers using chiral supercritical fluid chromatography
(SFC). Starting
reagents for this synthesis were sourced commercially using the LabNetwork
(www.labnetwork.com), which is a website that permits one to search for
chemical suppliers
for inputted structures/chemical names. There are numerous suppliers listed on
LabNetwork
for the starting compound, Compound 1 (e.g. Apollo Scientific Ltd., Stockport,
UK), and for
Compound 3-A (e.g. Astatech Inc., Bristol PA, USA) and Compound 5-A (e.g.
Atlantic
Research Chemicals Ltd., Bude, UK). Scheme 2
N
DPPA, DBU PPh3, H20
OH
DMF THF
N3
417% CI 77 4%
CI CI CI
1 2
CI
SI 3-A
CIJII'
NCS IBX
CH3CN NANH ACN, NH3 H20
NH2
i 89 8% a 4 a 40
c
3
Nr":z1
CI
NH
N
NANH C(9 ci 5
N10
HO 0-A
SFC
N NH
COI, THF
CI 4024 2% 19a CI
N CI N
N
N 0 N 0
11
NH(NNH
S
CI CI Ci ci
6a 6b
CI CI
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CA 3050553 2019-07-25
Products of Scheme 2:
Stereoisomer ("Stereoisomer 1", enantiomeric excess >97%): Liquid
Chromatography-Mass
Spectrometry (LC-MS): Liquid chromatography (LC) retention time (RT) = 2.516
minutes,
Mass spectrometry (MS; electrospray ionization, positive mode): m/z 537.1
[M+Hr, 559.1
[M+Na], 269.1 [M+2H]2 .
1H NMR (400 MHz, DMSO-d6) (ppm) 11.43 (s, 1H), 8.30 (s, 1H), 8.21 (d, J = 7.9
Hz, 1H),
7.93 (d, J = 7.8 Hz, 2H), 7.68 ¨ 7.56 (m, 3H), 7.56¨ 7.46 (m, 4H), 7.20 (s,
1H), 7.09 (d, J =
8.1 Hz, 2H), 6.97 (s, 1H), 5.96 (s, 1H), 4.33 (s, 1H), 4.19 (s, 1H). {NMR
probe temperature =
298.15 .
Opposite stereoisomer ("Stereoisomer 2", enantiomeric excess >97%): LC-MS:
Liquid
Chromatography RT = 2.516 minutes, Mass spectrometry (MS; electrospray
ionization,
positive mode): m/z 537.1 [M+H], 559.1 [M+Na], 269.1 [M+2H]2+.
1H NMR (400 MHz, DMSO-d6) 5 (ppm) 11.43 (s, 1H), 8.30 (s, 1H), 8.21 (d, J =
7.9 Hz, 1H),
7.93 (d, J = 7.8 Hz, 2H), 7.67 ¨ 7.56 (m, 3H), 7.50 (t, J = 6.5 Hz, 4H), 7.20
(s, 1H), 7.09 (d, J
= 8.2 Hz, 2H), 6.97 (s, 1H), 5.96 (s, 1H), 4.34 (s, 1H), 4.19 (s, 1H). {NMR
probe temperature
= 298.15 K} .
Presented NMR peaks are from using the "Auto Assignment" algorithm in
MestReNova
version 12 software (Mestrelab Research, Santiago de Compostela, Spain), which
picks peaks
in an inputted NMR spectrum, influenced by an inputted structure, which was
structure 19a in
Scheme 2. An invention embodiment is the use of a compound with LCMS and/or
NMR
features as presented above, for use in a method of treatment of the human or
animal body by
therapy, optionally for cancer treatment/amelioration/prevention/combat in a
subject. The
actual LCMS and NMR spectra are presented in Figure 32 herein, with further
information in
its legend.
The reaction scheme below, Scheme 3, is the same as Scheme 2 up until Compound
5. This
shared component isn't shown, just the point of divergence from Compound 5,
which comes
from using a different Compound 5-A, which is available from multiple
suppliers on
labnetwork.com (e.g. HE Chemical, Changzhou, Jiangsu, China).
Scheme 3
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CA 3050553 2019-07-25
CN
NH
5-A
1110 N)LNH
CI ci HO 0 N 0
COI, THF
N NH
CI CI CI *
CI
The reaction scheme below, Scheme 4, is the same as Scheme 2 up until Compound
5. This
shared component isn't shown, just the point of divergence from Compound 5,
which comes
from using a different Compound 5-A, which is available from multiple
suppliers on
5 labnetwork.com (e.g. HE Chemical, Changzhou, Jiangsu, China).
Scheme 4
CN
N
NH
NANH 5-A
CI CI HO 0 NZ
5 =COI, THF
ao N NH
CI CI c, is
CI
The reaction schemes below, Scheme 5 and Scheme 6, show only starting material
and
product because they use the same internal steps as Scheme 2 (not shown), but
with a
different starting material, a different Compound 1 than Scheme 2 and each
other, as shown
below, and thence different products produced, as shown. Compound 1 of Scheme
5 and
Scheme 6 are available from suppliers listed on labnetwork.com (e.g. Toronto
Research
Chemicals, Ontario, Canada).
Scheme 5
1110
D
D D
OH D CI DI
N NH
CI
1
CI
ci
CI
Scheme 6
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CA 3050553 2019-07-25
-;-,N
p
L/D
rs ts1
D N
OH
N NH
CI CI
1
CI CI 101
CI
The reaction scheme below, Scheme 7, is modified from Scheme 2, in order to
produce a
deuterated analogue, with deuterium in place of hydrogen on the chiral carbon.
The scheme is
provided for the purpose of illustrating the invention, and should not be
regarded in any
manner as limiting the scope or the spirit of the invention. This
illustrating, not limiting,
feature applies to all the compound synthesis schemes of this disclosure. The
starting
compound in the scheme below, Compound 1, is available from multiple suppliers
listed on
LabNetwork (e.g. Apollo Scientific Ltd., Stockport, UK). The 2nd compound,
Compound 2, is
also available from multiple suppliers listed on LabNetwork (e.g. Manchester
Organics Ltd.,
UK). So, one of the art can choose between these two starting options
depending on their
preference. Compound 2 tends to be more expensive than Compound 1 to buy and
the
Compound 1 to Compound 2 reaction shown gives high yield, and imidazole
(Compound 1A)
can be purchased cheaply (e.g. from Apollo Scientific Ltd., Stockport, UK),
thus starting
from Compound 1 as opposed to Compound 2 can be cheaper. In an alternative
embodiment
(not shown), Compound lA is replaced with Imidazole-13C,15N2 (CAS no: 1173018-
62-6;
available from suppliers on Labnetwork.com e.g. Meihezhiku(Wuhan)
Biotechnology Co.,
Ltd, China) in Scheme 7 to produce an alternative product of this invention,
isotopically
enriched for '3C and '5Ix1 at equivalent positions in the final product.
Scheme 7
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CA 3050553 2019-07-25
Br /---:-.-.1 iz--. -. -....1
r---NH N.......,N N 1
\`\..,.....N
?IA NaBD4 D DPPA, DBU
0 N ____... ________________ ==
DMF
K2CO3, DMF 0 OH
CI CI
1
Nn CI 2 ci ci 3 CI
_õ...N NCS
C
Of 5A
D PPh3, H20 I _____
...,..N
N3
THF
D =
NH2 CH3CN
CI CI
4
7:-.-...1 ----N
N..õ,..N 5 N 1
...,,....N
S NH hiJ7A
DA D...1,
,cI
N NH IBX
= N NH
H ACN, NH3 H20
6 7 a H HO 'LO
_________________________________________________________ =
CI CI 01 Cl 40 CD, THF
a tsl a
":õN ..:5,1=1
liz
=.,õ..N r---1- -/----- 1._
N.....-N N 0 N
D..,.. SFC I, N 0 N 0
D), .õ.u)..,
H 40 Fe/N NH 40 s N NH
H H
ci N)NH 8 c, 401 c, ci 40, c, ci 0,
CI CI CI
Products of Scheme 7:
Stereoisomer ("Stereoisomer A", enantiomeric excess >97%): LC-MS: LC retention
time
(RT) = 2.685 minutes, MS (electrospray ionization, positive mode): m/z 538.1
[M+H], 560.1
[M+Na], 269.6 [M+2F1]2+. High Resolution Mass Spectrometry (HRMS): Liquid
Chromatography-Time of Flight (LC-TOF) MS (electrospray ionization, positive
mode): LC
Retention Time (RT) = 0.166 minutes, m/z 538.0745928061 [M+H], m/z
560.0600137508
[M+Na], m/z 576.0250917093 [M+K], molar percent deuterium incorporation at
chiral
carbon = 99.13%.
IHNMR (400 MHz, DMSO-d6) a 11.48 (s, 1H), 8.33 (s, 1H), 8.25 (dt, J = 7.8, 1.5
Hz, 1H),
7.96 (dt, J = 7.7, 1.5 Hz, 1H), 7.92 (s, 1H), 7.71 ¨7.60 (m, 3H), 7.60¨ 7.49
(m, 4H),
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CA 3050553 2019-07-25
7.23 (s, 1H), 7.14 (s, 2H), 7.00 (s, 1H), 4.38 (d, J = 14.1 Hz, 1H), 4.23 (s,
1H). {NMR probe
temperature = 301 K).
Opposite stereoisomer ("Stereoisomer B", enantiomeric excess >97%): LC-MS: LC
retention time (RT) = 2.685 minutes, MS (electrospray ionization, positive
mode): m/z 538.1
[M+H]+, 560.1 [M+Na], 269.6 [M+2H]2+. HRMS: LC-TOF MS (electrospray
ionization,
positive mode): LC RT = 0.163 minutes, m/z 538.0727757864 [M+H], m/z
560.0513502753
[M+Na], m/z 576.0327248583 [M+K]+, molar percent deuterium incorporation at
chiral
carbon = 99.14%.
'H NMR (400 MHz, DMSO-d6) 5 11.46 (s, 1H), 8.32 (s, 1H), 8.24 (d, J = 7.9 Hz,
1H),
.. 7.95 (d, J = 7.7 Hz, 2H), 7.70 - 7.59 (m, 3H), 7.59 - 7.49 (m, 4H), 7.21
(s, 1H), 7.12 (d, J =
7.9 Hz, 2H), 6.99 (s, 1H), 4.36 (d, J = 13.9 Hz, 1H), 4.22 (s, 1H). {NMR probe
temperature =
300.7K}.
Presented NMR peaks come from using the "Auto Assignment" algorithm in
MestReNova
version 12 software (Mestrelab Research, Santiago de Compostela, Spain), which
picks peaks
in an inputted NMR spectrum, influenced by an inputted structure, which was
structure 8 in
Scheme 7. However, after automatic assignment, I manually made a single change
for the 2"d
stereoisomer: I changed the number of hydrogens at 7.95 ppm from 1H to 2H. I
executed this
change by manually extending the integration line (this manipulation
automatically changes
the MestReNova program's integration calculation method from "peak" to "sum")
in this
7.95 ppm region for the 2nd stereoisomer, making it the same length as the
integration line in
the same region for the 1st stereoisomer, which then rendered/integrated this
region with 2H
as compared to 1H, the same as for the 1st stereoisomer in this region (by
"Auto Assignment"
algorithm: 1st stereoisomer has peaks at 7.96 [1H] and 7.92 [1H] = 2H in 7.95
ppm region).
.. This change in integration increases the nuclide count to the correct
number for the structure
(18H). This manipulation is best explained in pictures and so please refer to
Figures 33L and
33M, which both show the 1st stereoisomer in upper panel, 2nd stereoisomer in
lower panel,
wherein Figure 33L shows how this region was automatically integrated and
Figure 33M
shows how I manually extended the integration line of the 2"d stereoisomer to
match the
.. length of the 1St stereoisomer, so that both now integrate to 2H in this
region.
On another issue, please note that for the 2nd stereoisomer, its LC-MS, HRMS
and NMR are
not completely aligned: to explain, LC-MS, FIRMS and 1H NMR were performed and
the
NMR showed a high amount of impurities (spectrum not shown), further
purification steps
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CA 3050553 2019-07-25
were taken, and the 1H NMR was repeated (spectrum shown herein), and a 13C NMR
also, all
showing a much lower, acceptable level of impurity. However, LC-MS and HRMS
were not
repeated. But the LC-MS and HRMS are clear enough to identify the [M+ion]
species
needed, all be it with some additional species/impurities also observed that
aren't in the LC-
MS and HRMS spectra for the lst stereoisomer.
An invention embodiment is the use of a compound with LC-MS and/or NMR
features as
presented above, for use in a method of treatment of the human or animal body
by therapy,
optionally for cancer treatment/amelioration/prevention/combat in a subject.
The actual LC-
MS and NMR spectra are presented in Figure 29 herein, with further information
in its
legend.
The separated stereoisomers of structure 19a in Scheme 2 have a 'H NMR peak at
5.96 ppm
= from the hydrogen attached to their chiral carbon. The present
stereoisomers, those produced
by Scheme 7, have this hydrogen replaced with a deuterium and thence this 5.96
ppm peak is
observed absent in their 'H NMR spectra. Indeed, aside from this absence all
these
stereoisomers have an incredibly similar 1H NMR spectra. The 1H NMR spectra of
hydrogen
vs. deuterium on chiral carbon stereoisomers are compared in Figure 34.
Compound 5 in Scheme 7 is of the form of Compound 1, the starting compound, in
the
molecule synthesis embodiments of [Pl] (presented in its "Process of
Preparation" section),
BUT with the exception that is deuterated on its chiral carbon. This
deuterated form can be
substituted into the synthesis schemes described in [Pl] to produce deuterated
molecules,
with deuterium on their chiral carbon, which are componentry to the present
invention, and in
a non-limiting embodiment, one of more of these new compositions of matter are
used as
anti-cancer medicines. Solvents, temperatures, pressures, and other reaction
conditions may
readily be selected by one of ordinary skill in the art. Starting materials
are commercially
available or can be readily prepared by one of ordinary skill in the art using
known methods.
Further methods to synthesize Compound 5, of Scheme 7 above, are given below
in Scheme
8.
Scheme 8
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CA 3050553 2019-07-25
DPPA, DBU PPh H20,
=
DMF THF
OH N3 NH2
1
CI CI CI CI CI CI
IH/D exchange H/D exchange
NO NO NO
D DPPA, DBU, D PPh H20 D Further
OH
DMF N3 THF " synthesis
4 2
ci cs CI
(deuterium can be incorporated at other positions also, not shown, depending
on the particular deuteration reaction used)
Compound 1 in Scheme 8 is available from multiple suppliers listed on
LabNetwork (e.g.
Apollo Scientific Ltd., Stockport, UK). Conducting Scheme 8, the aim is to
obtain a higher
degree of deuterium incorporation on the chiral carbon than natural abundance:
for
Compound 4 ultimately, and for Compound 1 first if the lower arm of the
synthesis route is
used. In both cases, the greater the deuterium incorporation, the better.
Deuterium
incorporation at other positions of each molecule is permissible and within
the scope of the
invention, as is elevated deuterium incorporation only at the chiral carbon.
Reactions
described in [L, H, G, K, M, .11, J2, J3, I, F, S] deuterate (herein defined
as replace hydrogen
with deuterium) the a-carbon to a secondary alcohol and so the chiral carbon
of Compound 1.
Reactions described in [A, B, P, El, E2, F] deuterate the a-carbon to primary
amines, thence
can deuterate the chiral carbon of Compound 4. Reactions described in [N]
deuterate sp3
carbons, thence can deuterate the chiral carbon of Compounds 1 and 4.
Reactions described
in [01, 021 can deuterate the I3-carbon to phenyl groups and so can deuterate
the chiral
carbon of Compounds 1 and 4. Reactions described in [R1, R2, Q I, Q2] can
deuterate widely,
upon aromatic and alkyl molecular components, and thence can deuterate the
chiral carbon of
Compounds 1 and 4. Reactions described in [D] deuterate the 13-carbon to
tertiary amines,
thence can deuterate the chiral carbon of Compounds 1 and 4. The teaching of
[D] is
.. especially preferred for use in the present context. Whichever option(s) is
chosen, solvents,
temperatures, pressures, and other reaction conditions may readily be selected
by one of
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CA 3050553 2019-07-25
ordinary skill in the art. The level of deuteration can be modulated by
modulating the reaction
time: greater deuterium incorporation by longer reaction time. One can do
multiple cycles of
one or more of these reactions until the desired level of deuterium
incorporation occurs,
monitored by 1H and/or 2H NMR (e.g. deuterium incorporation quantified by
decrease of 1H
NMR integral intensity at specified position(s) compared to starting material)
and/or mass
spectrometry. Some of the reactions cited herein use commercially available
catalysts e.g.
10% Pd/C catalyst [01, 02, Q I, R1, R2], and/or Pt/C catalyst [R1, R2, Ql,
Q2], or shvo
catalyst [D], or RuC12(P(Ph)3)3 (CAS no: 15529-49-4) [A], or 5% Ru/C catalyst
[K], or Ru-
macho catalyst [G, M], all available from Sigma-Aldrich. Others teach, or cite
literature
teaching, how to prepare the catalyst to use. The aforementioned list of
synthesis options, to
make Compound 4 in Scheme 8, is not exhaustive. A person skilled in the art
will know how
to find further options. For example using computational tools, including
artificial
intelligence (AI, non-limiting e.g. [222, 223, 224]), to search the chemical
reaction
literature/databases, e.g. (non-limiting) the Reaxys or CAS databases, and
their own skill in
the art to find, plan and prioritise synthesis routes. 2-(1H-imidazol-1-y1)-1-
phenylethanamine
is commercially available on LabNetwork and can be deuterated at its chiral
carbon (and
optionally at other positions also) by one or more of the aforementioned
methods disclosed
herein for deuterating the chiral carbon of 1-(2,4-dichloropheny1)-2-(imidazol-
1-
yl)ethanamine. Before or after deuteration, its phenyl group can be (non-
limiting) alkylated,
halogenated, or CF3 added (non-limiting example: at the 1UPAC 2,5 positions),
at desired
position(s) by methods well known to those of the art. Then it can be a
starting compound in
the synthesis schemes of [Pl] and used to produce deuterated compositions of
matter that are
componentry to this invention, which in non-limiting embodiments, are used
singly or in a
combination in anti-cancer therapy, in an animal or human. Alternatively, the
final products,
rather than starting materials, of the synthesis schemes of [Pl] can be
deuterated, to produce
deuterated compositions of matter that are componentry to this invention,
which in non-
limiting embodiments, are used singly or in a combination in anti-cancer
therapy, in an
animal or human. Reactions described in [A, B, El, E2, F] can deuterate the a-
carbon to
secondary amines, and thence the chiral carbon of Compound 19a, in Scheme 2
presented
previously, and the chiral carbon of other molecules with the scaffold of
[P1], as presented in
the abstract of [P1]. These compounds can also be deuterated at their chiral
carbon, and in
further embodiments at further or other position(s), by reactions described in
[N], which
deuterate sp3 carbons. And/or by reactions described in [01, 02], which can
deuterate the 3-
carbon to phenyl groups. And/or by reactions described in [R1, R2. Q1, Q2],
which deuterate
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CA 3050553 2019-07-25
aromatic and alkyl molecular components. And/or by reactions described in [D],
which can
deuterate the 13-carbon to tertiary amines. Some of these reactions are
stereoretentive [F, N.
El, E2] and thus can be used, optionally, after stereoisomer enrichment.
Others are not, e.g.
[Q1, Q2], and so should be used before any enantiomeric excess (ee) enrichment
step. All
patents and papers cited by the present disclosure, and their supplementary
materials, are
herein incorporated by reference, and are componentry, to the present
disclosure.
The reaction scheme below, Scheme 9, is the same as Scheme 7 up until Compound
7. This
shared component isn't shown, just the point of divergence from Compound 7,
which comes
from using a different Compound 7A, which is available from multiple suppliers
on
labnetwork.com (e.g. HE Chemical, Changzhou, Jiangsu, China).
Scheme 9
CN
I
NH
- 7A
r
1- cHDOI THO NN F. 0
IX<N NH
CI CI CI
CI
The reaction scheme below, Scheme 10, is the same as Scheme 7 up until
Compound 7. This
shared component isn't shown, just the point of divergence from Compound 7,
which comes
from using a different Compound 7A, wherein three different options of
Compound 7A are
shown and the three resultant products, wherein Compound 7A(i), 7A(ii) and
7A(iii) are all
available from multiple suppliers on labnetwork.com (e.g. all available from
Fluorochem,
Hadfield, Derbyshire, UK).
Scheme 10
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CA 3050553 2019-07-25
Br Br
7A(i)
0 NN0
HO
THF NNH
CI CI H110
CI
N 07A(ii)
NH
X*lZN NH Ho
CDI, THF N) NH
CI CI
7
CI CI
CI
CI
7A(Iii)
0 0
HO
D.)t,
COI, THFµ N NH
CI CI
CI
The reaction scheme below, Scheme 11, shows only starting material and product
because it
uses the same internal steps as Scheme 7, but with a different starting
compound, as shown
below (available from suppliers listed on labnetwork.com e.g. Matrix
Scientific, Columbia,
SC, USA), and thence a different product is produced, as shown.
Scheme 11
N
k
40 0 N
D)L.
CI 1 N NH
CI
CI
The reaction scheme below, Scheme 12, shows only starting material and product
because it
uses the same internal steps as Scheme 7(starting from its Compound 2), but
with a different
starting compound, as shown below (available from suppliers listed on
labnetwork.com e.g.
Vitas-M Laboratory, Champaign, IL, USA), and thence a different product is
produced, as
287
CA 3050553 2019-07-25
shown. Product shown is the (predicted, MarvinSketch software [Chemaxon,
Hungary])
predominant tautomer.
Scheme 12
,-N
H C
3 ¨N+
HaC¨tri
NH 0
0
N N
CI 4
CI CI
CI 0
CI
The reaction scheme below, Scheme 13, differs from Scheme 2 in order to
produce
methylated analogues, with methyl in place of hydrogen on the chiral carbon.
The starting
compound and imidazole are both available from Apollo Scientific Ltd.,
Stockport, UK and
other suppliers listed on labnetvvork.com. In an alternative embodiment (not
shown),
Compound IA is replaced with Imidazole-13C,15N2 (CAS no: 1173018-62-6;
available from
suppliers on Labnetwork.com e.g. Meihezhiku(Wuhan) Biotechnology Co., Ltd,
China) in
Scheme 13 to produce an alternative product of this invention, isotopically
enriched for 13C
and 'Nat equivalent positions in the final product.
Scheme 13
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CA 3050553 2019-07-25
Br fz"-----1 r----1
f N N
N 11"\
N.=,. NH CH3 HnSO4
MeMgBr
K2CO3, DMF 0 = OH DH3CN
THF
CI i CI
CI CI CI CI
2 3
/---:----1
NCS
N
CH3 ,,, HCI
--=- CH3 a
N--e
4 CI 5 CI CH3 CN
CN
N.õ....,N Isk\,,...,N` so
CH3S CH3NH
--11-,õ,õ IBX
a 1,,i4 .,.. ACN, NH3 -H2 6 a H N.11...NH HO 0
6
CI CI a c,= ci a CD, THF
7
CI CI
--- --- ---
CH.31 SFC . cH3N 0
),, H, 0
N NH 40 s N., NH 0 Fi NH
H F I
CI 8 CI (110 CI CI 100 CI CI ao
Products of Scheme 13:
Stereoisomer ("Stereoisomer a", enantiomeric excess >97%): LC retention time
(RT) =
2.536 minutes, MS (electrospray ionization, positive mode): m/z 551.0 [M+Hr,
573.0
[M+Na], 276.0 [M+2H]2+.
1H NMR (400 MHz, DMSO-d6) 8 (ppm) 11.37 (s, 1H), 7.89 (dt, J = 7.6, 1.5 Hz,
1H), 7.80 (d,
J = 7.8 Hz, 1H), 7.65 -7.56 (m, 3H), 7.55 -7.47 (m, 2H), 7.47 - 7.37 (m, 3H),
7.22 (d, J =
8.4 Hz, 2H), 7.05 (s, 1H), 6.95 (s, 1H), 6.91 (s, 1H), 4.88 (d, J = 13.6 Hz,
1H), 4.76 (d, J =
13.7 Hz, 1H), 1.70 (s, 3H). {NMR probe temperature = 298.2 K}
Opposite Stereoisomer ("Stereoisomer p", enantiomeric excess >97%): LC
retention time
(RT) = 2.540 minutes, MS (electrospray ionization, positive mode): m/z 551.1
[M+H], 573.0
[M+Na], 276.1 [M+2H]2+;
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CA 3050553 2019-07-25
Presented NMR peaks come from using the "Auto Assignment" algorithm in
MestReNova
version 12 software (Mestrelab Research, Santiago de Compostela, Spain), which
picks peaks
in an inputted NMR spectrum, influenced by an inputted structure, which was
structure 8 in
Scheme 13. However, after automatic assignment, I manually made a single
change: I
changed the number of hydrogens at 7.05 ppm from OH to 1H by manually changing
its
integration value from 0.41 to 0.5 (which is rounded to 1). This change in
integration
increases the nuclide count to the correct number for the structure (21H).
On another issue, please note that the presented LC-MS and NMR are not
completely
aligned: the LC-MS was conducted before, and the shown NMR after, HPLC
purification.
Prior NMR, before the HPLC purification, showed many impurities present, and
so the NMR
was repeated after HPLC purification (from which peaks are shown). This rd NMR
was only
performed for one of the stereoisomers.
An invention embodiment is the use of a compound with LC-MS and/or NMR
features as
presented above, for use in a method of treatment of the human or animal body
by therapy,
optionally for cancer treatment/amelioration/prevention/combat in a subject.
The actual LC-
MS and NMR spectra are presented in Figure 35 herein, with further information
in its
legend.
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CA 3050553 2019-07-25
Compound 5 in Scheme 13 is of the form of Compound 1, the starting compound,
in the
molecule synthesis embodiments of [P11 (presented in its "Process of
Preparation" section),
BUT with the exception that is methylated on its chiral carbon. This
methylated form can be
substituted into the synthesis schemes described in [Pl] to produce methylated
molecules,
with methyl on their chiral carbon, that are componentry to the present
invention, and in a
non-limiting embodiment, one of more of these new compositions of matter are
used as anti-
cancer medicines. Solvents, temperatures, pressures, and other reaction
conditions may
readily be selected by one of ordinary skill in the art. Starting materials
are commercially
available or can be readily prepared by one of ordinary skill in the art using
known methods.
In some embodiments, Scheme 13 is implemented with each intermediate purified
by HPLC,
especially the last 3 intermediates, wherein this yields a lower fraction of
impurit[y/ies] in the
final product. Two alternatives for a step in Scheme 13:
CN
CH3NH
N NH
HO 0 CH3N 0
CI ci T3P,THF
7 N NH
ci
CI 8 CI io
OR CI
CN
CI CI
CH3aNH
NNH N
HO 0 cH3N
o HATU,THF..
7 N NH
ci CI 8 CI
CI
The reaction scheme below, Scheme 14, differs from Scheme 2 in order to
produce
fluorinated analogues, with fluorine in place of hydrogen on the chiral
carbon.
Scheme 14
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CA 3050553 2019-07-25
f--z-.1 r-----t- Cl A /--,=-1
-N
N N.,...-N Nr.--1 03- N...-
....-N S
DPPA, DBU. PPh3, H20 NCS IBX
DMF .
OH th THF
NH2 CH3CN NANH ACN, NH3=1120
H
CI 1 CI CI 2 CI
CI CI CI 4 CI io
3
Selectfluor, K2S208, cl
MeCN/H20, heat (e.g. 80 C)
OR
Selectfluor,
Fe(acac)2 catalyst
MeCN
NH OR
5-A 1-:"---1
0 NANH HO __ 0 N\,.......N Selectfluor,
H .
N 0 9-fluorenone catalyst
CI 5 CI 0 CDI, THF .
__11 MeCN, visable light
(light bulb)
N¨NH OR
a H Selectfluor,
Cl 6 Cl 40 xanathone catalyst
MeCN, blacklight (360 nm)
CI OR
Selectfluor,
1,2,4,5-tetracyanobenzene (TCB) catalyst,.
MeCN, UV (302 nm) light
OR
TREAT=HF (+ optionally AgF)
[Mn(salen)Cl] catalyst
PhI0, MeCN, heat (e.g. 50 C)
OR
N-fluorobenzenesulfonimide (NFSI)
tetrabutylammonium decatungstate (TBDAT) catalyst
N NaHCO3, MeCN, hf (A = 365 nm)
v
.- ,--
U fliIP
Iµ1\,___N
N
RN NNN 0
F.,..4._ SFC N 0 N 0
N NH
H '`N H H"---NH 40 R N NH
CI 7 ci 40
c, ci 0 ci a a
ci
ci ci
Solvents, temperatures, pressures, and other reaction conditions may readily
be selected by
one of ordinary skill in the art. Starting materials are commercially
available (e.g. refer
www.labnetwork.com) or can be readily prepared by one of ordinary skill in the
art using
known methods. In alternative invention embodiments the fluorination step is
implemented
with an earlier or later intermediate than that shown in Scheme 14. A method
to fluorinate a
benzylic carbon in a structure of Scheme 14 using a Fe(acac)2 [225-226], or 9-
fluorenone
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CA 3050553 2019-07-25
[227], or xanathone [228], or 1,2,4,5-tetracyanobenzene (TCB) [229] or
[Mn(salen)C1] [230],
or tetrabutylammonium decatungstate (TBDAT) [231], or no [232] catalyst is
componentry to
this invention, optionally with the judicious (to someone of the art) addition
of a protecting
group(s) before, and removal after, the fluorination step e.g. (non-limiting)
protecting the OH
group, optionally with TMS, if fluorinating Compound 1. Alternative
fluorination methods
can be found in the literature, e.g. (non-limiting) refer [233] or [234].
Compound 3
(compound with the NH2 group) in Scheme 14 is of the form of Compound 1, the
starting
compound, in the molecule synthesis embodiments of [Pl] (presented in its
"Process of
Preparation" section). Its fluorinated form, fluorinated on its chiral carbon,
optionally by a
benzylic carbon fluorination reaction disclosed herein, can be substituted
into the synthesis
schemes described in [P1] to produce fluorinated molecules, with fluorine on
their chiral
carbon, that are componentry to the present invention, and in a further non-
limiting
embodiment, one of more of these new compositions of matter are used as anti-
cancer
medicines.
The following scheme can be substituted into Scheme 14 to produce
alternatively
halogenated invention embodiments. In other invention embodiments a
halogenation step is
implemented with an earlier or later intermediate of Scheme 14 than that shown
below.
NBS
MeCN, fluorescent lamp
OR <,1=1
N-bromosuccinimide (NBS)
SePPh3 40
Na0C1
CDCI3
Mn5(salen)C1
OR
N 0 N 0 TBACI, DCM, N2 N 0
Br_x, BBr4
OR
N NH CCI4 N NH TCCA N NH
OR DBDMH Cl6 ClH Cu(OAc)2=1120
CI CI a CI CI
40
NHPI, CH7C1,
ZrCI4
.CH2C12, ambient light
CI CI Cl
DIH or 3-ITMH catalyst N
o-NO2C6H4CO2H
NHPI, CH2Cl2
NN
N 0
I
N NH
CI CI
c,
A method to chlorinate [235-236], or brominate [237-240], or iodinate [241] a
benzylic
carbon in a structure of Scheme 14 is componentry to this invention,
optionally with the
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CA 3050553 2019-07-25
judicious (to someone of the art) addition of a protecting group(s) before,
and removal after,
the halogenation step.
Scheme 15
CH3
CI rt.,
I-% N"-- NH (NH
(NH
..., '-I
CI C
lb 1)TEA N.--
I . -
1 2) AcOH, HCI NaBD4N-- D DPPA, DBU.
0 OH DMF
T-NH
CI 2 CI CI 3 CI
\fµr
D PPh3, H20 c NH CI 5b
N3 THE
,
N
D NC.S
CI 4 CI
NH2 CH3CN
DA
eNH
CI CIeNH N-- S
N NH IBX N.--- NH I. 7b
J1ICkH ACN, NH3.H2 N--
0.. D u
CI 6 CI al -'NH
H HO 0 ,
CI ci a CDI, THF
7
7N --_õ,=N
T-NH
e
\µNr N 0 NH (NH
isr N 0
0).., _S_FS li-.,... . D).LN
N NH rNNH i N NH
H H10 CI OH,
tio
CI 8 CI 0 CI a a
ci ci ci
5
Scheme 16
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CA 3050553 2019-07-25
a cH3
eNH (NH
NI-- N--
a CI CI
H2S0,4,
lb 1) TEA 0 MeM9Br. OH CH3CN
CI 1 CI THF
2) AcOH, Ha
a3
rNli 2
(NH NCS
\N--
CH3 NCI N--- i
0 +. r .......4 3 (10 5b
=11¨f CH3 NH2
CI 4 CI CI '
(NH CIS 5 CI CH3CN
N-- CH
3 õ eml
, la CN
N--11.-, NH IBX N CH NH
3 H 11111WIL
NH N..
ACN, NH3 .H2O )L,
H
CI CI ao H HO 0 .
6
CI 7 CI a CDI, THF
CI õN ,- N "N
...- --.
CI
NH
eNH H r (-NH
N"--- C N 0 \N--
SFC __________________ . PH3 N 3,...tL.
N NH S N NH N.-- C., F1N 0
R ''N NH
H H H
CI 8 CI ao ci ci a ci ci 40
CI CI CI
For Scheme 15 and Scheme 16, starting materials are available from suppliers
listed on
labnetwork.com. For example: Compound 1 (e.g. Apollo Scientific Ltd.,
Stockport, UK),
Compound lb (e.g. ChemScene, Monmouth Junction, NJ, USA), Compound 5b (e.g.
Astatech Inc., Bristol PA, USA) and Compound 7b (e.g. Atlantic Research
Chemicals Ltd.,
Bude, UK). For Scheme 17 below, starting materials are available from
suppliers listed on
labnetwork.com: e.g. Compound lb from ChemScene, Monmouth Junction, NJ, USA;
e.g.
Compound 5b from HE Chemical, Changzhou, Jiangsu, China.
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CA 3050553 2019-07-25
Scheme 17
CI ¨CH3 (NH
rl N-.";''= NH N¨
..., NH20H
lb 1)TEA
CI CI . 0 Zn, HCI
1 2) AcOH, HCI
CI
CI 2 CI
r NH NH
411D _... r
N¨ Li N-- NCS S. NH3, HgOs
NH2 CH3CN N NH
H
CI CI CI 4 CI ao
rNH CN
CI
N¨
NH 5b ("NH
N ANH N¨
N 0
HO 0 II SFC,
H
CDI THF
CI 5 CI * ' - N---"NH
H
CI 6 CI Op
CI
CI
(¨NH
eNNH
N¨ N 0 N¨ N 0
H Ji õM., ji.s.
$ N NH
H H
CI CI 5 CI CI 0
CI CI
For Scheme 18, starting materials are commerically available from suppliers
listed on
labnetwork.com: Compound 1 (Toronto Research Chemicals, Ontario, Canada),
Compound 4
and Compound 7 (Astatech Inc., Bristol PA, USA).
Scheme 18
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CA 3050553 2019-07-25
NJ
7--:--1- /----
N.---N D N....-N D N.....-N1 D
D D DPPA DBU D D PPh3, H20. D D
OH DMFJJJ(N3 THF
NH2
CI i CI CI 2 CI CI 3 CI
,-N
.,-- -->N Nr-L-1
....-N D
D D
CS2, Mel
________________ = NH2
H3C 4 0
I 0 CI 3 CI
H3C¨S 5 S¨CH3
NH2
r----1- r=1¨
D D 1 7 CI, D D I
N S¨CH3 N NH
H 8 H
CI CI CI CI 40
8
CI
For Scheme 19, starting materials are commerically available from suppliers
listed on
labnetwork.com: Compounds 1, 3, 9, 11 (Toronto Research Chemicals, Ontario,
Canada).
Scheme 19
-
NJr-,----1 o o /--,--1
N...¨N D Nt,,,N D Et01,1{, 1µ1,,,..N D
D D Mn 2 D Et0 3 0E,t D 0
'..
OH 0 4 OEt
CI 1 CI CI 2 CI CI CI
N/------1 r---1
N,..-NI
Pd/C, H2 D 0 NH4OH D D 0 TFAA
5 OEt 6 NH2 :=N
CI CI CI CI
=/=----1 P-------1
µ....N D 9
Et0H D D NH CI 0 __ .
=N
7 8 OEt =N
CI CI =N CI .. CI
NH2 Nr'---/¨
D
D D N 0
011 I
N 0
D 1 _ CI 12 NH
OEt CI CI 0
ci ci
cl
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Reaction Scheme 2 in more detail
In this section, numbers in square brackets are CAS numbers.
Nr-- N17----
...,N1
DPPA, DBU Isl
______________________ D.
OH DMF N3
CI CI CI CI
1 2
I. Charge Compound 1 [24155-42-8] (10.0 g, 1.00 X by weight) and DPPA [26386-
88-9]
(10.7 g, 1.07 X by weight) in DMF [68-12-2] (50 mL, 5.00 X by volume).
2. Charge DBU [6674-22-2] (5.90 g, 0.59 X by weight) into the mixture under
nitrogen
atmosphere at 0 C, then stir for about 15 min.
3. After that, take the reaction mixture to 20-30 C and stir for 24 h.
4. TLC (DCM/Me0H = 10/1, Rf = 0.6) show raw material remains, LC-MS (SM: RT =
0.729
min, Product: RT = 0.822 min) indicates reaction mixture has desired product.
5. The reaction mixture is extracted with Et0Ac (100 mL x 3).
6. The combined organic phase is washed with aq.NaC1 (100 mL), dried with
Na2SO4,
filtered and concentrated in vacuum to get the residue.
7. The residue is purified by column chromatography (DCM/Me0H = 1/0 to 100/1).
8. Compound 2 (4.82 g, 41.7%, 95% purity) is obtained as yellow oil, which is
confirmed by
HNMR and HPLC (Retention Time = 1.919 min).
Compound 2: 1H NMR (CDC13, 400 MHz) 6 (ppm) 7.47 (d, J = 2.2 Hz, 1H), 7.41 (s,
1H),
7.33 - 7.28 (m, 1H), 7.26 - 7.23 (m, 1H), 7.06 (s, 1H), 6.92 (s, 1H), 5.26
(m,1H), 4.23 (m,
1H), 4.02 (m, 1H).
Ni---- hill
,.,,Isl \,_..,...N
PPh3, H20
N3 THF NH2
CI CI CI CI
2 3
1. Charge Compound 2 (3.00 g, 1.00 X by weight) and PPh3 [603-35-0] (8.40 g,
2.80 X by
weight) in H20 (1 mL, 0.33 X by volume) and THF [109-99-9] (10 mL, 3.33 X by
volume).
2. Stir at 70 C for 24 h.
3. Check the reaction by LC-MS (SM: RT = 0.822 min, Product: RT = 0.299 min),
it shows
Compound 2 is consumed completely and desired mass is detected.
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4. The reaction mixture is concentrated under reduced pressure to remove TI-
IF.
5. The reaction mixture is added with H20 (30 mL), adjust pH to 3 with con.HC1
and washed
with Et0Ac (30 mL).
6. After that collect water phase, adjust pH to 13 with IN aq.Na0H, then exact
with Et0Ac
(30 mL x 2).
7. The combined organic phase is washed with aq.NaC1 (30 mL), dried with
Na2SO4, filtered
and concentrated in vacuum to obtain the product.
8. Compound 3 (2.23 g, 77.4%, 94.5% purity) is obtained as a yellow liquid,
which is
confirmed by HNMR and HPLC (Retention Time = 0.658 min).
Compound 3: 1H NMR (CDC13, 400 MHz) 5 (ppm) 7.48 - 7.37 (m, 3H), 7.30 - 7.26
(m, 1H),
7.06 (s, 1H), 6.91 (s, 1H), 4.72 (m, 1H), 4.21 (m, 1H), 3.91 (m, 1H).
rsirs
CI
3-A Mi Ncs N )L NH
NH2 CH3CN CI CI 40
CI c,
CI
3 4
1. Charge compound 3 (2.00 g, 1.00 X by weight) and compound 3-A [2131-55-7]
(1.30 g,
0.65 X by weight) in CH3CN [75-05-8] (30 mL, 15.00 X by volume).
2. Stir at 20-30 C for 12 h.
3. Check the reaction by TLC (DCM/Me0H = 10/1, Rf = 0.7) and LC-MS (SM: RT =
0.299
min, Product: RT = 0.949 min), desired product is detected, TLC indicated
Compound 3 is
consumed completely.
4. The reaction mixture is concentrated under reduced pressure to remove CH3CN
to obtain
the residue.
5. The residue is purified by column chromatography (DCM/Me0H = 1/0 to 50/1).
6. Compound 4 (2.95 g, 89.8%, 99.7% purity) is obtained as a white solid,
which is
confirmed by HNMR and HPLC (Retention Time = 2.521 min).
Compound 4: 1H NMR (DMSO-d6, 400 MHz) 5 (ppm) 9.81 (s, 1H), 8.54 (d, J = 7.5
Hz, 1H),
7.64 (s, 1H), 7.55 (s, 1H), 7.53 - 7.48 (m, 1H), 7.45 - 7.41 (m, 114), 7.38
(d, J = 8.6 Hz, 2H),
7.28 (d, J=8.4 Hz, 2H), 7.11 (s, 1H), 6.94 (s, 1H), 6.03 (s, 1H), 4.46 - 4.34
(m, 1H), 4.31 -
4.22 (m, 1H).
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NH
NANH IBX NA NH
CI CI C H 3C N, NH3 H20 CI CI
*
CI CI
4 5
1. Charge Compound 4(1.63 g, 1.00 X by weight) and IBX [61717-82-6] (1.18 g,
0.75 X by
weight) in CH3CN [75-05-8] (16 mL, 10.00 X by volume) and NH3.H20 [1336-21-6]
(8 mL,
5.00 X by volume).
2. Stir at 20-30 C for 18 h.
3. Check the reaction by LC-MS (SM: RT = 0.949 min, Product: RT = 1.050 min)
and HPLC
(SM: RT = 2.535 min, Product: RT = 1.757 min), desired product is detected.
4. The reaction mixture is extracted with Et0Ac (20 mL x 3).
5. The combined organic phase is washed with aq.NaC1(20 mL), dried with
Na2SO4, filtered
and concentrated in vacuo to obtain the crude product (1.0 g, white solid),
which is used into
next step directly.
N
116
NH
NNH 5-A
N 0
HO 0
N NH
CI 5 ci 40 COI, THF
CI
19a io
ci
CI
1. Charge Compound 5-A [1877-72-1] (0.36 g, 0.36 X by weight) and CDI [530-62-
1] (0.40
g, 0.36 X by weight) in THF [109-99-9] (20 mL, 20.00 X by volume), stir at 20-
30 C for 3 h.
2. Charge Compound 5 into the reaction mixture, stir at 20-30 C for 14 h.
3. Check the reaction by LC-MS (SM: RT = 1.050 min, Product: RT = 1.347 min)
and TLC
(DCM/Me0H = 10/1, Rf = 0.75), desired mass is detected. TLC indicates the raw
material is
consumed completely.
4. The reaction mixture is concentrated under reduced pressure to remove THF.
5. The reaction mixture is extracted with Et0Ac (20 mL x 3)
6. The combined organic phase is washed with aq.NaC1(20 mL), dried with
Na2SO4, filtered
and concentrated in vacuo to obtain the residue.
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7. The residue is purified by column chromatography (DCM/Me0H = I/O to 100/1)
and
chiral SFC {column: AD (250 mm*50 mm, 10 gm); mobile phase: [0.1%NH3H20 IPA];
B%:
28%-28%, 3.5 min} separating the stereoisomers of Compound 19a: Stereoisomer
1(0.17 g,
SFC RT = 2.35 min) is obtained as a white solid. Stereoisomer 2 (0.15 g, SFC
RT = 2.69
min) is obtained as a white solid.
Reaction Scheme 7 in more detail
In this section, numbers in square brackets are CAS numbers.
Br 1 1A ,õN1
0 0
K2CO3, DMF
CI CI CI CI
1 2
1. Charged Compound 1 [170894-53-8] (15.0 g, 55.9 mmol) and DMF [68-12-2] (150
mL)
into the reactor at 25 C under N2.
2. Charged K2CO3 [584-08-7] (23.2 g, 167 mmol) into the reactor at 25 C.
3. Charged imidazole [288-32-4] (11.4 g, 167 mmol) into the reactor at 25 C.
4. The mixture was stirred at 25 C for 3 h.
5. TLC (petroleum ether:ethyl acetate = 5:1, Rf = 0.0) showed raw material was
consumed.
6. The mixture was quenched with water (100 mL).
7. The aqueous phase was extracted with ethyl acetate (100 mL x 2). The
combined organic
phase was washed with brine (50 mL), dried with anhydrous Na2SO4, filtered and
concentrated in vacuum.
8. The residue was purified by silica gel chromatography (column height: 250
mm, diameter:
100 mm, 100-200 mesh silica gel, petroleum ether/ethyl acetate=20/1, 3/1) to
give Compound
2 (13.0 g, 38.8 mmol, 69.3% yield, 76.1% purity) as yellow solid.
Compound 2: 1H NMR (CDC13, 400 MHz) Es (ppm) 7.55 (d, J = 8.0 Hz, 1H), 7.50
(s, 1H),
7.35-7.37 (dd, J = 8.0 Hz, 8.0 Hz, 1H), 7.09 (d, J = 8.0 Hz, 111), 6.93 (s,
1H), 5.32 (s, 2H).
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NaBD4
0 OH
CI CI CI CI
2 3
1. Charged Compound 2 (8.00 g, 31.3 mmol) and Me0H (40 mL) into the reactor at
25 C
under N2.
2. Charged NaBDa [15681-89-7] (1.31 g, 34.5 mmol) to the solution.
3. The mixture was stirred at 50 C for 3 h.
4. LCMS (product: RT= 0.914 min) showed raw material was consumed.
5. The reaction was quenched with water (100 mL).
6. The aqueous phase was extracted with ethyl acetate (50 mL x 2). The
combined organic
phase was washed with brine (50 mL), dried with anhydrous Na2SO4, filtered and
concentrated in vacuum.
7. The residue was purified by silica gel chromatography (column height: 250
mm, diameter:
100 mm, 100-200 mesh silica gel, petroleum ether/ethyl acetate=10/1, 0/1) to
give Compound
3 (5.50 g, 21.3 mmol, 67.9% yield) as white solid.
Compound 3: 1HNMR (CDCI3, 400 MHz) 5 (ppm) 7.57 (d, J = 8.0 Hz, 1H), 7.39 (m,
1H),
.. 7.28-7.30 (dd, J = 8.0 Hz, 8.0 Hz, 1H), 6.88 (d, J = 8.0 Hz, 2H), 4.20 (d,
J = 12 Hz, 1H), 3.84-
3.88 (d, J = 16 Hz, 1H).
DPPA, DBU
OH DMF N3
Ci CI Ci CI
3 4
1. Set up a reactor R-1. (Note: R-1 is a 100 mL three-necked bottle)
2. Charged Compound 3 (4.50 g, 17.4 mmol) and DPPA [26386-88-9] (4.80 g, 17.4
mmol,
3.78 mL), DMF (22.5 mL) into the reactor R-1 at 25-30 C.
3. Charged DBU [6674-22-2] (2.65 g, 17.4 mmol, 2.63 mL) into the mixture under
nitrogen
atmosphere at 0-5 C, then the mixture was stirred for about 15 min.
4. After that, the reaction mixture was stirred for 24 h at 20-30 C.
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5. TLC (DCM/Me0H = 10/1, SM Rf = 0.55, product Rf = 0.72) showed raw material
was
consumed, LCMS (product: RT = 0.764 min) and HPLC (product: RT = 1.893 min)
indicated
reaction mixture has desired product.
6. The reaction mixture was extracted with Et0Ac (40 mL x 3).
7. The combined organic phase was washed with brine (40 mL), dried with
Na2SO4, filtered
and concentrated in vacuum to get the residue.
8. The residue was purified by column chromatography (SiO2, dichloromethane:
methanol =
1/0 to 100/1) to give Compound 4 (3.80 g, crude) as yellow oil.
Compound 4: 1H NMR (CDCI3, 400 MHz) 8 (ppm) 7.50 (s, 1H), 7.44 (s, 1H), 7.35
(d, J =
2.0 Hz, 1H), 7.33 (d, J = 2.0 Hz, 1H), 7.29-7.30 (m, 1H), 6.94-7.10 (m, 1H),
4.25 (d, J = 14.4
Hz, 1H), 4.04 (d, J = 14.4 Hz, 1H).
Nf="---1-- 1::----1
PPh3, H20
N3 THF NH2
CI CI CI CI
4 5
1. Set up a reactor R-1. (Note: R-1 is a 100 mL three-necked bottle)
2. Charged Compound 4 (3.80 g, 13.4 mmol), PPh3 [603-35-0] (10.5 g, 40.2
mmol), THF
[109-99-9] (12.6 mL) and H20 (1.26 mL) into the reactor R-1 at 25-30 C.
3. Stirred at 65-70 C for 16 h.
4. Checked the reaction mixture by TLC (dichloromethane: methano1=10:1, SM: Rf
= 0.54,
product: Rf= 0.45) and LCMS (product: RT = 1.253min) showed that reactant was
consumed
completed.
5. The reaction mixture was concentrated under reduced pressure to remove THF.
6. The reaction mixture was added with H20 (38 mL), adjusted pH to 3 with con.
HC1 and
washed with Et0Ac (38 mL).
7. The water phase was adjusted pH to 13 with IN aq. NaOH, then exacted with
Et0Ac (38
mL x 2).
8. The combined organic phase was washed with brine (38 mL), dried with
Na2SO4, filtered
and concentrated in vacuum to give Compound 5 (2.60 g, crude) as a yellow
liquid.
Compound 5: 1H NMR (CDC13, 400 MHz) 8 (ppm) 7.44 (d, J = 14 Hz, 1H), 7.41 (s,
1H),
7.29 (d, J = 2.0 Hz, 1H), 7.27 (d, J = 1.2 Hz, 1H), 7.07 (s, 1H), 6.92 (s,
1H), 4.21 (d, J = 14
Hz, 1H), 3.92 (d, J = 14 Hz, 1H).
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CI a 5A
NCS
N NH
NH2 CI CI CH3CN
CI CI (10
6
CI
1. Charged Compound 5(2.50 g, 9.72 mmol), Compound 5A [2131-55-7] (1.65 g,
9.72
mmol) and MeCN (37.5 mL) into the reactor at 25 C.
5 2. The mixture was stirred at 25 C for 12 h.
3. TLC (dichloromethane: methano1=10:1, SM: Re= 0.5, product: Re= 0.2) showed
raw
material was consumed completed.
4. The mixture was concentrated in vacuum.
5. The residue was purified by silica gel chromatography (column height: 250
mm, diameter:
100 mm, 100-200 mesh silica gel, petroleum ether/ethyl acetate=3/1, 0/1) to
give Compound
6 (3.60 g, 8.44 mmol, 86.7% yield) as white solid.
Compound 6: 1H NMR (CDC13, 400 MHz) ö (ppm) 9.08 (s, 1H), 7.48 (d, J = 2.0 Hz,
1H),
7.33 (d, J = 8.0 Hz, I H), 7.25-7.28 (m, 1H), 7.16 (d, J = 2.0 Hz, 1H), 7.00
(s, 1H), 6.88 (d, J =
8.0 Hz, 1H), 6.72 (s, 1H), 4.46 (dd, J = 14.4 Hz, 14 Hz, 1H), 1.84 (s, 1H).
NH
DA DA
N NH __ IBX N NH
ACN, NH3 H20 ci
CI CI CI
6 7
CI CI
1. Charged Compound 6 (3.00 g, 7.03 mmol) and MeCN (30 mL) into the reactor at
25 C.
2. Charged IBX [61717-82-6] (2.17 g, 7.73 mmol) into the reactor at 25 C.
3. Charged NH3.H20 [1336-21-6] (13.6 g, 116 mmol, 15 mL, 30% purity) into the
reactor at
25 C.
4. The mixture was stirred at 25 C for 12 h.
5. LCMS (product: RT = 0.717 min) showed raw material was consumed completed.
6. The mixture was filtered and the filter cake was concentrated in vacuum to
give Compound
7 (2.50 g, 6.10 mmol, 86.8% yield) as white solid.
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Compound 7: 1H NMR (CDC13, 400 MHz) 5 (ppm) 7.87 (d, J = 8.0 Hz, 11-1), 7.67
(s, 1H),
7.55-7.60 (m, 2H), 7.37-7.41 (m, 2H), 7.23 (s, 1H), 7.07-7.09 (m, 1H), 6.92
(s, 1H), 4.33 (dd,
J = 14.4 Hz, 14 Hz, 1H).
N
, N /
Ni----- IW
NH 7A r---
D)1 N
, HO 0 s\N
N NH N 0
H D
CM, THF
CI CI 0 N NH
H
CI CI io
7 8
CI
CI
1. Charged Compound 7 (538 mg, 3.66 mmol), CDI [530-62-1] (593 mg, 3.66 mmol)
and
THF [109-99-9] (30 mL) into the reactor at 25 C.
2. The solution was stirred at 25 C for 3 h.
3. Charged compound 7A [1877-72-1] (1.50 g, 3.66 mmol) into the reactor at 25
C.
4. The mixture was stirred at 25 C for 12 h.
5. LCMS (SM: RT = 0.730 min, product: RT = 1.030 min) showed the raw material
was
consumed/completed.
6. The mixture was concentrated in vacuum.
7. Seperating stereoisomers of Compound 8: the residue was purified with Prep-
HPLC
(column: DAICEL CHIRALPAKAD-H (250 mm*30 mm, 5 pm); mobile phase: [0.1%
NH3H20 IPA]; B%: 44%-44%, 6.45 min) to give Stereoisomer A (0.025 g, 45.8
ttmol, 1.25%
yield, 98.8% purity) as white solid and Stereoisomer 13 (0.025 g, 45.5 mot,
1.24% yield,
98.1% purity) as white solid.
Reaction Scheme 13 in more detail
In this section, numbers in square brackets are CAS numbers.
r ____________________
B Nr------1
N-/----1-
_..NH
0 b.
K2003,DMF 0
01 01
01 01
1 2
1. Compound 1 (10.0 g, 37.3 mmol) was added in DMF (100 mL) at 25 C under
N2.
2. K2CO3 [584-08-7] (15.4 g, 111 mmol) was added to the solution.
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3. Imidazole [288-32-4] (7.62 g, 111 mmol) was added.
4. The mixture was stirred at 25 C for 3 h.
5. TLC (petroleum ether: ethyl acetate = 5:1, Rf = 0.35) showed raw
material was
completed.
6. The mixture was poured into water (100 mL) and extracted with ethyl
acetate (100
mLx 2).
7. The combined organic phase was washed with brine (100 mL), dried with
anhydrous
Na2SO4, filtered and concentrated in vacuum.
8. The residue was purified by silica gel chromatography (100-200 mesh
silica gel,
Petroleum ether/Ethyl acetate = 20/1, 3/1).
9. Compound 2 (7.20 g, 14.4 mmol, 38.7% yield, 51.2% purity) was obtained
as a
yellow solid.
Compound 2: 1H NMR (CDC13, 400 MHz) 8 (ppm) 7.55 (d, J = 8.00 Hz, 1 H), 7.48-
7.49 (m,
2 H), 7.34-7.36 (m, 1 H), 7.09 (s, 1 H), 6.90 (s, 1 H), 5.31 (s, 2 H).
LC-MS: RT = 1.02 min, m/z 254.9 [M+H].
MeMgBr
0 THF OH
CI CI CI CI
2 3
1. MeMgBr (3.0 M, 91.4 mL, 274 mmol) was added to the reactor and cooled
to 0 C
under N2.
2. A solution of Compound 2 (7.00 g, 27.4 mmol) in THF (70 mL) was added.
3. The mixture was stirred at 25 C for 12 h.
4. TLC (petroleum ether: ethyl acetate = 5:1, Rf = 0) showed raw material
was
completed.
5. The reaction was quenched with water (100 mL) and extracted with ethyl
acetate (100
mL x 2).
6. The combined organic phase was washed with brine (100 mL), dried with
anhydrous
Na2SO4, filtered and concentrated in vacuum.
7. Compound 3 (9.70 g, crude) was obtained as a yellow solid.
Compound 3: 1H NMR (CDC13, 400 MHz) 8 (ppm) 7.63 (d, J = 6.36 Hz, 1 H), 7.31-
7.44 (m,
3 H), 7.10 (d, J = 5.50 Hz, 1 H), 6.70 (s, 1 H), 4.34-4.52 (m, 2 H), 1.62 (s,
3 H).
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LC-MS: RT = 1.05 min, m/z 270.9 [M+H]t
Nrn,
CH3CN 0
N OH H2SO4
CI CI CI CI
3 4
1. To a mixture of Compound 3 (3.00 g, 11.0 mmol) in CH3CN (40 mL) was
added
H2SO4 [7664-93-9] (15 mL, 281mm01) in one portion at 20 C under N2.
2. The mixture was stirred at 40 C for 10 h.
3. LC-MS (product: RT = 0.63 min; SM: RT = 0.72 min) showed the reaction
was
completed.
4. The mixture was poured into ice-water (w/w = 1/1) (200 mL) and adjust to
pH = 7
with 10% aq.Na0H.
5. The aqueous phase was extracted with DCM (100 mL x 2).
6. The combined organic phase was washed with brine (100 mL), dried with
anhydrous
Na2SO4, filtered and concentrated in vacuum.
7. Compound 4 (3.00 g, crude) was obtained as a yellow solid.
1H NMR (CDCI3, 400 MHz) 6 (ppm) 7.32-7.35 (m, 2 H), 7.18 (m, 2 H), 6.94 (s, 1
H), 6.72
(s, 1 H), 5.99 (s, 1 H), 5.09 (d, J = 13.7 I-1z, 1 H), 4.53 (d, J = 13.7 Hz, 1
H), 1.57 (s, 3 H).
dTh
0 Con HCI
NH2
CI CI CI CI
4 5
1. A mixture of Compound 4(2.70 g, 8.65 mmol) in Con.HC1 (12 M, 50 mL, 600
mmol)
was stirred at 100 C for 40 h.
2. LC-MS (product: RT = 0.32 min; SM: RT = 0.67 min) showed the reaction
was
completed.
3. The mixture was adjusted to PH = 7 with 10% aq.Na0H.
4. The aqueous phase was extracted with ethyl acetate (100 mL x 2).
5. The combined organic phase was washed with brine (100 mL), dried with
anhydrous
Na2SO4, filtered and concentrated in vacuum.
6. The residue was used into next step without purification.
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7. Compound 5 (3.00 g, crude) was obtained as a yellow solid.
CI At
"PP NCS
1IIitN
5A
11.=
NH2 CH3CN A NH CI CI SI
CI CI
CI
6
1. A mixture of compound 5 (3.00 g, 11.1 mmol) and compound 5A [2131-55-7]
(1.88
5 g, 11.1 mmol) in CH3CN (30 mL) was stirred at 20 C for 12 h.
2. LC-MS (product: RT = 0.95 min; SM: RT = 0.32 min) showed the reaction
was
completed.
3. The mixture was poured into water (20 mL) and extracted with ethyl
acetate (30 mL x
2).
4. The combined organic phase was washed with brine (30 mL), dried with
anhydrous
Na2SO4, filtered and concentrated in vacuum.
5. The residue was purified by silica gel chromatography (100-200 mesh
silica gel,
CH2C12/Me0H = 200/1, 50/1).
6. Compound 6 (1.80 g, 4.09 mmol, 36.8% yield) was obtained as a yellow
solid.
Compound 6: 1H NMR (CDC13, 400 MHz) 8 (ppm) 7.50 (s, 1 H), 7.34 (s, 1 H), 7.19-
7.26 (m,
4 H), 6.94-7.06 (m, 4 H), 6.23 (s, 1 H), 6.58-6.59 (m, 1 H), 4.35 (d, J = 13.0
Hz, 1 H), 1.50 (s,
3H).
NH
NANH _______________________________________ N A NH
IBX
CI CI 40 CH3CN, NH3=1120 CI CI 410
CI CI
6 7
1. To a mixture of compound 6(1.80 g, 4.09 mmol) in CH3CN (20 mL) was added
IBX
[61717-82-6] (1.26 g, 4.50 mmol) and NH3.H20 (9.0 mL, 70.1 mmol) in one
portion at 20 C
under Nz.
2. The mixture was stirred at 20 C for 3 h.
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CA 3050553 2019-07-25
3. LC-MS (product: RT = 0.73 min; SM: RT = 0.95 min) showed the reaction
was
completed.
4. The mixture was poured into water (20 mL) and extracted with ethyl
acetate (20 mL x
2).
5. The combined organic phase was washed with brine (20 mL), dried with
anhydrous
Na2SO4, filtered and concentrated in vacuum.
6. The residue was used into next step without purification.
7. Compound 7 (1.50 g, crude) was obtained as a yellow oil.
7.N
40 CN
NNH TA
HO 0 N 0 N 0
40 COI, THF
CI CI
7 N NH N NH
CI CI C ao
CI I
R stereoisomer/ R stereoisomer/
S stereoisomer_A CI S stereoisomer_B CI
1. To a mixture of compound 7A [1877-72-1] (278 mg, 1.89 mmol) in CH3CN (20
mL)
was added CDI [530-62-1] (306 mg, 1.89 mmol) in one portion at 25 C under N2.
2. The mixture was stirred at 25 C for 3 h.
3. Then Compound 7 (800 mg, 1.89 mmol) was added into the mixture, and
stirred at
25 C for 12 h.
4. LC-MS (product: RT = 0.73 min; SM: RT = 1.03 min) showed the reaction
was
completed.
5. The mixture was poured into water (20 mL) and extracted with ethyl
acetate (10 mL x
2).
6. The combined organic phase was washed with brine (10 mL), dried with
anhydrous
Na2SO4, filtered and concentrated in vacuum.
7. The residue was purified by silica gel chromatography (100-200 mesh
silica gel,
CH2C12/Me0H = 200/1, 50/1) to give 400 mg compound R stereoisomer / S
stereoisomer,
and further purified by SFC (SFC condition: column: DAICEL CHIRALPAK AD-H
(250mm*30mm, 5 pm); mobile phase: [Neu-Me0H]; B%: 60%-60%, 5 min).
8. Compound R stereoisomer / S stereoisomer_A (80.0 mg, 144 pmol, 15.3%
yield,
100% e.e.) was obtained as an off-white solid.
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9. Compound R stereoisomer / S stereoisomer_B (80.0 mg, 144 1.1mol,
15.3% yield,
98.9% e.e.) was obtained as an off-white solid.
EXAMPLE (H)
Summary of Formula (II)
This invention embodiment relates to compounds having the formula:
Formula (H)
(R1)m
\Aa/ (R7)d
$k
(Cvv)w Zõ
Rr x / v `.(C')g
/ ,
(Cf)f/<
including
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CA 3050553 2019-07-25
(R1)m
Aa (R7)d
k
Nh
)s
(Cw)w µz n
Rr v k
/ ' x ________ L
G.(Cf)fi
including
(R1)m
(R7)d
Aa,
N "
Z
ik (Cg)g
,4)w
Rr,T tõY X
Gi
Gi
(Cf)f L
including
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CA 3050553 2019-07-25
(Rilm
A/(R7)d a,
%
N(h)h
R2 Z
A
E\ ---1 4N........f___EA \ (cg)g
I
,,I, ik
R6
L
R3N2 -..,... L
R4 (Cf)f L
Bb
\
(R5)n
including
(R1)m
( R7
)q
R2
E\
ifsit .....*Rr=-..Tt.....-Y-..,N
N¨Z=
R35 /
11 R6
---
R4 N L
¨Bb
\
(R5),,
or pharmaceutically-acceptable salts, solvates, hydrates or prodrugs thereof,
wherein:
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CA 3050553 2019-07-25
GI is, independently at each point of use, aryl or heteroaryl;
G2 is, independently at each point of use, N or CH;
L is independently at each point of use alkyl, or substituted alkyl, or
deuterated alkyl, or
aminoalkyl, or thioalkyl, or alkoxy, or halogen, or haloalkyl, or haloalkoxy,
or hydroxyalkyl,
or any atom or isotope permitted by valence (including any accompanying
hydrogens by
valence e.g. (non-limiting) OH, NH2, SH, SiH3, PH2 etc.), for example
hydrogen, or
deuterium, or fluorine;
A is nitrogen (N), or N , or carbon;
E is absent, or alkyl, or substituted alkyl, or deuterated alkyl, or
aminoalkyl, or thioalkyl, or
alkoxy or any atom or isotope permitted by valence (including any accompanying
hydrogens
by valence e.g. (non-limiting) OH, NH2, SH, SiH3, PH2 etc.), for example
hydrogen, or
deuterium or fluorine;
Cf, Cg, Ch and C"' are each independently selected from a single bond, 0, S,
Se, Nle, PRv,
BRv, C(Rv)2 or Si(Rv)2, wherein each le is independently selected from a
constituent group
of L (defined earlier);
x, w, f, g, h are independently selected to be 0, 1, 2 or 3;
d is a selected integer between 0 and 7;
k, s and sk are independently selected to be 0, 1, 2, or 3;
The 5-sided ring structure is attached by any one of its available ring atoms,
and none, one or
two of its bonds can be a double bond, for example at locations shown by the
"single or
double bond" symbol;
RI and R5 are attached to any available carbon atom of phenyl rings A and Bb,
respectively,
and at each occurrence are independently selected from PH2, OH, SH, hydrogen,
deuterium,
alkyl, substituted alkyl, trifluoromethoxy, halogen, haloalkyl, cyano, nitro,
OR8, NR8R9,
C(=0)R8, CO2R8, C(=0)NR8R9, NR8C(=0)R9, NR8C(=0)0R9,
S(0)0R9, NR8S02R9, S02NR8R9, cycloalkyl, heterocycle, heterocyclo, aryl, and
heteroaryl,
and/or two of RI and/or two of R5 join together to form a fused benzo Ting;
R2, R3 and R4 are independently selected from E (defined earlier), hydrogen,
or deuterium, or
alkyl, or deuterated alkyl, and substituted alkyl, or one of R2, R3 and R4 is
a bond to R, T or Y
and the other of R2, R3 and R4 are independently selected from hydrogen,
alkyl, and
substituted alkyl;
Z and Y are independently selected from C(=0),-0O2-, -SO2---, -CH2-,
-CH2C(=0)-, and -C(=0)C(=0)-, or Z may be absent;
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CA 3050553 2019-07-25
R and T are selected from ¨CH2--, ¨C(=0)¨, and ¨CH RCH2)p(Q)]¨, wherein Q is
NRioRti, 0R10 or CN;
R6 is selected from thienyl, alkyl, alkenyl, substituted alkyl, substituted
alkenyl, aryl,
substituted aryl, cycloalkyl, heterocyclo, heteroaryl and aryl optionally
substituted with a
lower aliphatic group or one or more functional groups selected independently
from the
group consisting of¨NH2, -OH, phenyl, halogen, (Cl-C4)alkoxy or -NHCOCH3;
R7 is selected from L (defined earlier), PH2, OH, SH, hydrogen, alkyl,
substituted alkyl,
alkenyl, substituted alkenyl, aminoalkyl, halogen, haloalkyl, cyano, nitro,
keto (=0), hydroxy,
alkoxy, alkylthio, C(=0)H, acyl, CO2H, alkoxycarbonyl, carbamyl, sulfonyl,
sulfonamidyl,
cycloalkyl, heterocycle, heterocyclo, aryl, and heteroaryl;
R8 and R9 are independently selected from hydrogen, alkyl, substituted alkyl,
C2-4 alkenyl
optionally substituted, cycloalkyl, heterocycle, heterocyclo, aryl, and
heteroaryl, or R8 and R9
taken together to form a heterocycle or heterocyclo or heteroaryl, except R9
is not hydrogen
when attached to a sulfonyl group as in S02R9;
R10 and RI I are independently selected from hydrogen, alkyl, and substituted
alkyl;
m and n are independently selected from 0, 1, 2 and 3
o, p and q are independently 0, 1 or 2; and
r and t are 0 or 1.
Preferred compounds of Formula (H)
Preferred methods are to use, and preferred compounds are, compounds with the
following
formula, or pharmaceutically-acceptable salts, solvates, hydrates or prodrugs
thereof,
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CA 3050553 2019-07-25
(R1),,
R2 Aka, (R7)q
E\
A N¨Z
' t N
R3-'5 NZ R6
R4
(R5),,
including
(R1),,
(R7)q
R2 AaZ
N¨Z
t R6
R4
¨13-b
(R5)õ
wherein:
L is hydrogen, or deuterium, or methyl, or hydroxyalkyl, or fluorine;
A is nitrogen (N), or N+, or carbon;
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CA 3050553 2019-07-25
E is absent, or alkyl, or substituted alkyl, or deuterated alkyl, or
aminoalkyl, or thioalkyl, or
alkoxy or any atom or isotope permitted by valence (including any accompanying
hydrogens
by valence e.g. (non-limiting) OH, NH2, SH, SiH3, PH2 etc.), for example
hydrogen or
deuterium;
R1 and R5 are attached to any available carbon atom of phenyl ring Aa and
phenyl ring Bb,
respectively, and at each occurrence are independently selected from hydrogen,
deuterium,
alkyl, aralkyl, aminoalkyl, halogen, cyano, nitro, hydroxy, alkoxy,
trifluoromethoxy,
alkylthio, NH2, NH(alkyl), N(alkyl)2, C(=0)H, acyl, CO2H, alkoxycarbonyl,
carbamyl,
sulfonyl, sulfonamide, cycloalkyl, heterocycle, heterocyclo, aryl, and
heteroaryl, and/or two
.. of RI and/or two of R5 join together to form a fused benzo ring;
R2, R3 and R4 are independently selected from hydrogen and alkyl;
Z is ¨0O2¨, ¨S02¨, or is absent;
Y, R and T are selected from ¨CH2¨ and ¨C(=0) ¨,
R6 is selected from:
Cmalkyl or Ci_aalkenyl optionally substituted with up to three of halogen,
aryl and CO2Ci-
6a1ky1;
phenyl optionally substituted with up to three R12 and/or having fused thereto
a benzo-ring or
a five to six membered heteroaryl;
heteroaryl selected from thiophenyl, imidazolyl, pyrazolyl, and isoxazolyl
wherein said
heteroaryl is optionally substituted with up to two Ri2,
R7 is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted
alkenyl,
aminoalkyl, halogen, cyano, nitro, keto (=0), hydroxy, alkoxy, alkylthio,
C(=0)H, acyl,
CO2H, alkoxycarbonyl, carbamyl, sulfonyl, sulfonamide, cycloalkyl,
heterocycle,
heterocyclo, aryl, and heteroaryl;
R12 at each occurrence is independently selected from each other R12 from the
group
consisting of Ci_6alkyl, halogen, nitro, cyano, hydroxy, alkoxy, NHC(=0)alkyl,
¨0O2alky1,
¨S02phenyl, aryl, five to six membered monocyclic heteroaryl, and phenyloxy or
benzyloxy
in turn optionally substituted with halogen, hydroxyl, Ci_aalkyl, and/or
0(Ci_4a1ky1);
m and n are independently selected from 0, 1, 2 or 3; and
q is 0, 1 or 2; and
r and t are 0 or 1.
More preferred are compounds having the following formula, or pharmaceutically-
acceptable
salts, solvates, hydrates or prodrugs thereof,
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CA 3050553 2019-07-25
(R1),,
R2
\ N N-Z
\
R3 N R6
H L
11 (R5),,
wherein
RI and R5 are attached to any available carbon atom of phenyl ring Aa and
phenyl ring Bb,
respectively, and at each occurrence are independently selected from alkyl,
halogen, cyano, hydroxy, alkoxy, NH2, NH(alkyl), N(alkyl)2, C(0)H, acyl, CO2H,
,
alkoxycarbonyl, and/or two of Ri and/or two of R5 join together to form a
fused benzo ring;
R2, R3 and R4 are independently selected from hydrogen and lower alkyl;
Z is ¨0O2¨, ¨S02¨, or is absent;
R6 is selected from:
C i_aalkyl or Ci_aalkenyl optionally substituted with up to three of halogen,
aryl and CO2C i_
6alkyl;
phenyl optionally substituted with up to three R12 and/or having fused thereto
a benzo ring or
a five to six membered heteroaryl;
heteroaryl selected from thiophenyl, imidazolyl, pyrazolyl, and isoxazolyl,
wherein said
heteroaryl is optionally substituted with up to two RI2,
RI2 at each occurrence is independently selected from each other R12 from the
group
consisting of Ci_6 alkyl, halogen, nitro, cyano, hydroxy, alkoxy,
NHC(=0)alkyl,
¨0O2alkyl, ¨S02phenyl, aryl, five to six membered monocyclic heteroaryl, and
phenyloxy
or benzyloxy in turn optionally substituted with halogen, hydroxyl, C1-4
alkyl, and/or 0(C1-4
alkyl); and
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CA 3050553 2019-07-25
m and n are independently selected from 0, 1, or 2.
Even more preferred are compounds as immediately defined above wherein R6 is
selected
from Ci_aalkyl, trifluoromethyl, benzyl, C2_3alkenyl substituted with phenyl,
(R15),, (R17), (R17),
411 R16 , __ et, , ________
0
(R17)v (R17)v
N
wherein:
RI5 is halogen, alkyl, nitro, cyano, hydroxy, alkoxy, NHC(=0)alkyl, and/or two
RI5 groups
are taken together to form a fused benzo ring or a five to six membered
heteroaryl;
RI6 is selected from hydrogen, deuterium, halogen, alkyl, nitro, cyano,
hydroxy, alkoxy,
NHC(=0)alkyl, and phenyloxy or benzyloxy in turn optionally substituted with 1
to 3 of
hydrogen, deuterium, halogen, cyano, and Ci_aalkoxy;
RI7 is selected from alkyl, alkoxy, CO2C1_6alkyl, and SO2phenyl;
and u and v are independently 0, 1 or 2.
Most preferred compounds of Formula (II) are those having the formula:
R2
N N ¨Z \
R6
NH
=
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CA 3050553 2019-07-25
wherein
L is deuterium;
R2 is hydrogen or CH3;
Z is ¨0O2¨, ¨S02¨, or is absent; and
R6 is selected from the groups recited immediately above, most preferably
(R15)u
R16
Example embodiments of Formula (II)
Compounds from [8, 12], selected as specific anti-cancer therapeutics by the
invention of this
disclosure, selected because they inhibit the reverse, more than the forward,
mode of ATP
synthase. EC50 and IC50 used interchangeably. EC50 values for FIFO ATP
hydrolysis, and FiFo
ATP synthesis, in NADH-linked and NADPH-linked sub-mitochondrial (SMP) assays
respectively, sourced from [8], are presented. [8] refer to these ECso values
as IC50 values for
inhibiting FIFO ATP hydrolase (reverse mode) and FIF0 ATP synthase (forward
mode).
However, this in incorrect. Because, as identified by the invention of this
disclosure,
explained herein, although these molecules inhibit FiFo ATP hydrolase, their
reducing of FiFo
ATP synthesis is not (predominantly) because of inhibiting FiFo ATP synthase,
but by
uncoupling.
4y\
---NH 0 0
N\__ N-- N--S
./
H3C 113C
=
CI
CI
H3C CH3CH3
EC50 FiFo ATP hydrolase = 0.022 (pM) EC5c, F1F0 ATP hydrolase = 0.077 (pM)
EC, F1F0 ATP synthesis > 30 (pM) EC50FiFo ATP synthesis > 30 (pM)
EC 50 Ratio >1,364 EC Ratio >390
5o
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CA 3050553 2019-07-25
For all compounds: EC50 F1F0 ATP synthesis > 30 pAl
R6 RS imidazole ECK, FiFo ATP hydrolase ( M)
4-F-Ph SO2 5-y1 0.221
Ph SO2 5-y1 0.282
4-0H-Ph SO2 5-y1 0.667
4-0Me-Ph SO2 5-y1 0.077
2,5-di-CI-Ph SO2 5-y1 0.158
4-(AcNH)-Ph SO2 5-y1 2.981
4-CN-Ph SO2 5-y1 0.255 .4
IN1-.....R5
2-CI-4-CN-Ph 502 5-y1 0.939 -44 , . ' N
.,
3-NO2-Ph 502 5-y1 0.423 \
Naphth-1-y1 SO2 5-y1 0.338 R6
Thiophen-2-y1 SO2 5-y1 0.636
Benzofurazan-7-y1 SO2 5-y1 1.777
Quinolin-8-y1 SO2 5-y1 2.935
Bn SO2 5-y1 2.405
CF3 SO2 5-y1 0.077
4-t-Bu-Ph SO2 5-y1 0.008
41/
4-t-8u-Ph CH2 5-y1 2.138
4-t-Bu-Ph CH2 4-Me-5-y1 2.352
4-t-Bu-Ph SO2 2-y1 >10
4-F-Ph 502 2-Me-5-y1 9.623
4-F-Ph SO2 4-Me-5-y1 0.151
Further examples [12],
/-=--N 0 ,0 /-=---N 1110 , 0 ,=N 0
HN õ- NITS, H ,..IN N N-S' HN,
N 041h 0 N -,S' CH3
WIP- 1 . H3C
F Cl CH3
Further,
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CA 3050553 2019-07-25
N., õN N N¨,-,S
"---
CI
CI
=
Scheme Ha is a route for synthesizing Compound 31 [8], starting reagents are
commercially
available from multiple suppliers listing on the LabNetwork
(www.labnetwork.com) e.g.
Compounds 1, 7 and 9 are available from Astatech Inc., Bristol PA, USA,
Compound 2 from
Stru Chem, Wujiang city, China.
Scheme Ha
0
2
COOEt ., H COOEt
is NH 4 N
TFA, Et3Sill o.
NH2 toluene, 130 C, 18 h NH DCE, 0-25 C, 2 h
1
IIP
01 H
N LiAIH4
1N NaOH 0
4 .
NH2 HN 0 C, 18 h 0 THF, 25 C, 1Erh
111
COOEt NH
0 CI,.. //0 5
/IS Is a
H 0 7
ON Cl
N
TEA, DCM, 25 C, 3 h.-
NH
6
ii--NH 5 0
H N--r---- \ I/
SI H3C.--L
N ...N.,1H Ny,)\__ N¨S
9
--, HC N 0//
Ni= ,0 0 ,
NaBH(0/1/403, DCE, 25 C 1. Cl
0-S
8 . CI Ci
it, 31
Cl
321
CA 3050553 2019-07-25
Product of Scheme Ha, Compound 31:
LC retention time (RT) = 0.87 minutes, MS (electrospray ionisation, positive
mode): m/z
554.90 [M+Hr, 576.90 [M+Na], 278.90 [M+2H]2+, (all Observed m/z are within 0.3
Daltons of Expected: 555.14 [M+11] , 577.12 [M+Na] , 278.07 [M+211_72");
1H NMR (400 MHz, Methanol-d4) 8 (ppm) 8.35 (s, 1H), 8.14 - 8.06 (m, 1H), 8.04
(d, J = 2.1
Hz, 1H), 7.84 - 7.71 (m, 2H), 7.27 (dt, J = 22.4, 7.4 Hz, 4H), 7.16 (dd, J =
8.0, 5.6 Hz, 2H),
7.09 (d, J = 7.5 Hz, 2H), 7.03 (t, J = 7.4 Hz, 1H), 4.71 (d, J = 13.9 Hz, 1H),
4.37 (d, J = 14.2
Hz, 1H), 4.29(d, J = 14.2 Hz, 1H), 3.96 (dd, J = 19.7, 13.4 Hz, 2H), 3.17 -
3.10 (m, 1H),
2.78 (d, J = 12.8 Hz, 1H), 2.69 (ddd, J = 14.5, 9.7, 5.3 Hz, 11-1), 2.54 (dt,
J = 13.7, 8.6 Hz,
1H), 2.11 (s, 3H), 1.50 (s, 1H), 1.36 (dtd, J = 14.1, 9.1, 5.1 Hz, 1H) {NMR
probe temperature
= 297.9K).
1H NMR (400 MHz, Chloroform-d) 8 (ppm) 8.21 (s, 1H), 7.93 (d, J = 2.1 Hz, 1H),
7.82 (s,
1H), 7.65 (dd, J = 8.4, 2.1 Hz, 1H), 7.58 (d, J = 8.4 Hz, 11-1), 7.30 (s, OH),
7.26 (s, 2H), 7.20
(t, J = 7.3 Hz, 1H), 7.10 (d, J = 7.4 Hz, 2H), 7.07 - 6.98 (m, 2H), 4.60 (d, J
= 13.8 Hz, 1H),
4.32 (d, J = 14.1 Hz, 1H), 4.12 (dd, J = 24.1, 14.0 Hz, 2H), 3.82 (d, J = 12.8
Hz, 1H), 3.08 (s,
2H), 2.91 (d, J = 13.0 Hz, 1H), 2.76 - 2.64 (m, 1H), 2.49 (dt, J = 14.8, 8.2
Hz, 1H), 2.11 (s,
3H), 1.58 - 1.43 (m, 1H) {NMR probe temperature = 298.5 .
"C NMR (101 MHz, Methanol-d4) 8 (ppm) 149.14, 142.91, 140.73, 138.20, 134.62,
134.51,
132.72, 132.22, 130.62, 130.05, 129.75, 129.50, 129.39, 129.20, 128.48,
127.87, 127.01,
123.78, 121.75, 58.88, 53.98, 53.69, 48.11, 33.55, 31.58, 9.53 {NMR probe
temperature =
298.0 K1.
Presented NMR peaks come from using the "Auto Assignment" algorithm in
MestReNova
version 12 software (Mestrelab Research, Santiago de Compostela, Spain), which
picks peaks
in an inputted NMR spectrum, influenced by an inputted structure, which was
Compound 31
in Scheme Ha. However, for the 13C NMR spectrum, after automatic assignment, I
manually
intervened because the central Methanol-d4 solvent peak was observed at 49
instead of
expected 47.60 ppm. By my intervention, this 49 ppm Methanol-d4 solvent peak
was then
used/set as a "reference peak" in MestReNova which shifted all ppm values
accordingly (by
[49-47.60] ppm).
An invention embodiment is the use of a compound with LCMS and/or NMR features
as
presented above, for use in a method of treatment of the human or animal body
by therapy,
optionally for cancer treatment/amelioration/prevention/combat in a subject.
The actual
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CA 3050553 2019-07-25
LCMS and NMR spectra are presented in Figure 32 herein, with further
information in its
legend.
Compound 31, in Scheme Ha, can be deuterated at its chiral carbon, and in
further
embodiments at further or other position(s), by reactions described in [N],
which deuterate
sp3 carbons. And/or by reactions described in [R1, R2, Ql, Q2], which
deuterate aromatic
and alkyl molecular components. And/or by reactions described in [D], which
deuterate a and
13-carbons to tertiary amines. And/or by reactions described in [F, El, E2,
Exl], which
deuterate a carbons to tertiary amines. Alternatively, to produce a Compound
31
isotopologue, deuterated on its chiral carbon, and in further embodiments at
further or other
position(s), an intermediate in Compound 31 synthesis, presented in Scheme Ha,
can be
deuterated. For (non-limiting) example, Compound 6 in Scheme Ha can be
deuterated by
reactions described in [A, B, El, E2, F], which can deuterate the a-carbon to
secondary
amines. And/or by reactions described in [N], which deuterate sp3 carbons.
And/or by
reactions described in [R1, R2, Q I, Q2], which deuterate aromatic and alkyl
molecular
components. A deuterated Compound 6 can be inputted into synthesis schemes of
[P2], in
place of Compound 10 in Scheme III in the "Process of Preparation" section of
[P2], to make
deuterated isotopologues with the scaffold of [P2]. These are componentry to
the present
invention as new compositions of matter, and in non-limiting embodiments are
used singly or
in combination, optionally in co-therapy with an FDA and/or EMA approved
medicine(s)
and/or treatment(s), for example a licensed cancer treatment, as anti-cancer
therapeutics.
Throughout this disclosure, deuteration methods conveyed are illustrative
rather than
limiting. All stereoisomers of all the compounds of the present invention are
contemplated,
either in admixture or in pure or substantially pure form.
Scheme Jib
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CA 3050553 2019-07-25
o
2
COOEt ., H COOEt
4 N
NH2NH2 toluene, 130 C, 18 hi 0 Ill NH TFA, Et3SiH
DCE, 0-25 C, 2
1
SO H
IN NaOH N
0 Boc20
4 = =
NH2 HN 0 C, 18 h 0 DMAP, TEA,
THF
NH
COOEt 11, 5
111P Boc
I Boc
N I
Mel = N CH3 HCI, Me0H
1110 N......Bo0c t-BuOK, DMF
11101 o
6 N--õBoc
IP, 7
0 H 410,
N CH3 LiAl H4 c,,0
NH 0 THF, 25 C, 18 h H
N 0 01 S Cl
//
8
1110 cH3 CI
TEA, DCM, 25 C, 3 hi.
II 9 NH
r---NH 1110 0
H N.=-- \ ../y\\__ N¨S
N //
N
SI CH3 H3c .., --,12 NH N ,**/
H3C 0
r/
Rt.,- 0 0 0. CH3
...,,o- NaBH(OAc)3, DCE, 25 C Cl
U.-
11 c
c, ,
c, .
The reaction schemes below, Scheme Ilc, Scheme lid, Scheme Ile and Scheme Ill,
show
only starting material(s) and product because they use the same internal steps
as Scheme Ha
5 (not shown), but with a different starting material, a different Compound
1 in the case of
Scheme Hc, Scheme lid, Scheme He, or a different Compound 2 in the case of
Scheme Ilf,
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CA 3050553 2019-07-25
and thence different products are produced as shown. Starting compounds are
available from
suppliers listed on labnetwork.com: e.g. Compound 1 of Scheme lk and Scheme
Ile from
Fluorochem, Hadfield, Derbyshire, UK, e.g. Compound 1 of Scheme lid from J&W
Pharmlab LLC, Levittown, PA, USA, e.g. Compound 2 of Scheme Ilf from Arena
Chemical,
.. La Mure, France, Compound 1 of Scheme Ilf from Astatech Inc., Bristol PA,
USA.
Scheme lk
NH2
0'
NH2 H3C
1 CI
CI
Scheme lid
NH2 b¨NH
NS'
0
H3C ç 4110 ci
1
NH2 CI
Scheme Ile
40 NH2 ,0
N N-S
1 NH2 H3C /
CI
CI
Scheme Ilf
2
1110 ,
io NH2 COOEt 0
N 0
NH2 toluene, 130 C, 18h H3C
1 411 CI
CI
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CA 3050553 2019-07-25
The reaction schemes below, Scheme Hg and Scheme Hit are the same as Scheme Ha
up
until Compound 6. This shared component isn't shown, just the point of
divergence from
Compound 6, which comes from using a different Compound 7, wherein the
Compound 7
options shown in Scheme Hg and Scheme Ms are available from multiple suppliers
on
labnetwork.com (e.g. available from Fluorochem, Hadfield, Derbyshire, UK).
Scheme lig
ci
=
o=s=0
7CI
W.:7A 110
--(;)
N
NH TEA. DCM, 25 C, 3ChI.
8 N-1 H3C
9
H3C
6 o--=s=o NaBH(OAc)3, DCE, 25 C 40
c,
c,
Scheme Hh
\ /
IS 7 -NH WI= \ ir 1110
= H3c N=Nr.AN
TEA, DCM, 25 C, 3 h 110). LçJ 9
H3c
NH 8 0
6 NaBH(OAc)3, DCE, 25 C
The reaction schemes below, Scheme Hi, Scheme II j, Scheme Ilk and Scheme III
are the
same as Scheme Ha up until Compound 8. This shared component isn't shown, just
the point
of divergence from Compound 8, which comes from using a different Compound 9,
wherein
the Compound 9 options shown in Scheme Hi, Scheme I lj, Scheme Ilk and Scheme
III are
all available from multiple suppliers on labnetwork.com (e.g. all, except that
for Scheme Ill,
available from Fluorochem, Hadfield, Derbyshire, UK, Compound 9 from Scheme
III
available from Matrix Scientifjc, Columbia, SC, USA).
Scheme Ili
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CA 3050553 2019-07-25
H 14,---\ Tr-NH 110 ,P
Br-Cc,
Nõ\i,,,,,-õ.1.\\_, 11-7S
0 N 9
,.. Br N O'Ar
N _____________________ 0, _0
-
OS aBH(0A03. DCE, 25 C
'
8 . CI CI
CI
Scheme Hj
* H3C, 110 .
H N-:-"\ ,CH3 fi¨N ,0
=N--cs Br--4
9 Ny-,\\___N
Br
o'at
N0. o .
-
S - NaBH(OAc), DCE, 25 C
O'
8
CI
11 a
Cl
Scheme Ilk
HA .H N--=\- ,CH3 F-N ,0
1.-1:;k<4 N.,?,\._
1110 N 9 N O'Aia
I
N0, O W ci
-
S - NaBH(OAc),, DCE 25`C
O'
CI
8 41 CI
CI
Scheme III
H
N 110
N
N(;;;"--
\:=,,,o
N ¨
\ ,..ti . di CI
*S NaBH(00s,03, DCE, 25 C
0
CI
8 11 ci
a
Reaction Scheme Ha in more detail
In this section, numbers in square brackets are CAS numbers.
327
CA 3050553 2019-07-25
0
2 COOEt
H COOEt
NH2
NH2 toluene, 130 C, 18 II-7.
NH
1
3
A mixture of 2-(aminomethyl)aniline (2 g, 16.37 mmol, 1 eq) and ethyl 2-oxo-4-
phenyl-
butanoate (3.38 g, 16.37 mmol, 3.10 mL, 1 eq) in toluene (100 mL) was stirred
at 130 C for
18 h. TLC(PE:Et0Ac=1:1, rf=0.6) showed the starting material was consumed and
a new
.. spot was generated. The mixture was concentrated to the desired material
(4.8 g, yield:
94.5%) as a brown oil, which was used for next step without further
purification.
Compound 3: ill NMR (CDC13, 400 MHz) 6 (ppm) 7.25 - 7.16 (m, 3 H), 7.16 - 7.07
(m, 3
H), 6.96 (t, J=7.2 Hz, 1H), 6.83 (d, J=7.3 Hz, 1 H), 6.63 (dt, J=0.7, 7.4 Hz,
1 H), 6.52 (d,
J=7.9 Hz, 1 H), 4.22 - 4.08 (m, 2 H), 4.04 - 3.81 (m, 2 H), 2.83 - 2.66 (m, 1
H), 2.66 - 2.50
(m, 1 H), 2.13 -2.01 (m, 2 H), 1.21 (t, J=7.1 Hz, 3 H).
H COOEt TFA, Et3S1H
N
NH DCE, 0-25*C, 2 h .. NH2HN
COOEt
3 4
To a solution of ethyl 2-(2-phenylethyl)-3,4-dihydro-1H-quinazoline-2-
carboxylate (3.3 g,
10.63 mmol, 1 eq) in DCE (30 mL) at 0 C was added TFA (11.86 g, 104.00 mmol,
7.70 mL,
9.78 eq) and Et3SiH (2.08 g, 17.91 mmol, 2.86 mL, 1.68 eq). The resultant
solution was
stirred at 25 C for 2 h. The mixture was concentrated to dryness, the residue
was used for
next step without further purification. Compound 4: LC-MS: RT= 1.057 min, m/z
296.2 (M-
NH2)+.
1N NaOH
NH2HN 0 C, 18 h
0
COOEt NH
4 5
Ethyl 2[2-(aminomethypanilino]-4-phenyl-butanoate (4.5 g, 10.55 mmol, 1 eq,
TFA) was
dissolved in Me0H (50 mL), then the solution was adjusted pH=12-13 with IN
NaOH. The
resultant solution was stirred at 25 C for 16 h. The mixture was filtered to
give the desired
328
CA 3050553 2019-07-25
material, which was used for next step directly. Compound 5: LC-MS: RT= 0.879
min, m/z
289.1 (M+Na).
Compound 4: 1H NMR (CDC13, 400 MHz) 8 (ppm) 7.39 - 7.28 (m, 2 H), 7.26 - 7.18
(m, 3
H), 7.08 (t, J=7.8 Hz, 1 H), 6.91 (d, J=7.4 Hz, 1 H), 6.68 (t, J=7.4 Hz, 1 H),
6.54 (d, J=8.0 Hz,
1 H), 6.15 (br s, 1 H), 4.90 (dd, J=5.9, 16.2 Hz, 1 H), 4.36 (br t, J=6.5 Hz,
1 H), 3.89 (dd,
J=7.1, 16.3 Hz, 1 H), 3.52 (br s, 1 H), 2.98 - 2.84 (m, 1 H), 2.84 - 2.71 (m,
1 H), 2.44 - 2.24
(m, 1 H), 1.95 (m, 1 H).
LiAIH4
N
NH 0 THF, 25 C, 18 h
NH
To a solution of LiA1H4 (313.51 mg, 8.26 mmol, 1.1 eq) in THF (20 mL) was
added a
solution of 2-(2-phenylethyl)-1,2,4,5-tetrahydro-1,4-benzodiazepin-3-one (2.0
g, 7.51 mmol,
1 eq) in THF (20 mL). The resultant solution was stirred at 25 C for 18 h. The
reaction was
quenched with 1 mL water, and filtered. The filtrate was concentrated to
dryness. The residue
was purified by reverse chromatography (TFA) to give 2-(2-phenylethyl)-2,3,4,5-
tetrahydro-
1H-1,4-benzodiazepine (1.2 g, 4.13 mmol, 55.03% yield, 86.9% purity) was
obtained as a
brown oil.
LC-MS: RT= 0.668 min, m/z 253.3 (M+H)+.
NMR (CDC13, 400 MHz) 8 (ppm) 7.37 - 7.30 (m, 2H), 7.26 - 7.13 (m, 5H), 6.97 -
6.89
(m, 1H), 6.64 (d, J=7.8 Hz, 1H), 4.30 (d, J=14.1 Hz, 1H), 4.14 - 4.00 (m, 1H),
3.45 (br d,
J=11.7 Hz, 2H), 3.32 - 3.21 (m, 1H), 3.12 - 3.01 (m, 1H), 2.90 - 2.72 (m, 2H),
2.02- 1.89 (m,
2H).
CI,
S
7 op
N N 0
NH TEA, DCM, 25 C, 3 h C?'S/'
CI
6 8 a
To a solution of Compound 6 (220 mg, 871.79 mol, 1 eq) in DCM (5 mL) at 25 C
was
added TEA (363.50 mg, 3.59 mmol, 0.5 mL, 4.12 eq) and Compound 7 (214.04 mg,
871.79
mol, 1 eq). The resultant solution was stirred at 25 C for 2 h. LCMS showed
the starting
329
CA 3050553 2019-07-25
material was consumed and the desired MS was found. The mixture was
concentrated to
dryness, and the residue was purified by reverse chromatography (HCOOH) to
give
Compound 8 (170 mg, 362.36 gmol, 41.57% yield, 98.35% purity) as a colorless
oil.
Compound 8: LC-MS: RT= 1.058 min, m/z 461.1 (M-FH)+.
Compound 8: ill NMR (CDC13, 400 MHz) 7.72 (d, J=2.1 Hz, 1H), 7.46 - 7.41 (m,
1H), 7.40
- 7.36 (m, 1H), 7.34- 7.28 (m, 2H), 7.26- 7.21 (m, 1H), 7.20- 7.13 (m, 3H),
7.09 (dt, J=1.5,
7.6 Hz, 1H), 6.92- 6.84 (m, 1H), 6.50 (d, J=7.7 Hz, 1H), 4.59 -4.44 (m, 1H),
4.41 -4.26 (m,
1H), 3.61 (dd, J=2.1, 13.3 Hz, 1H), 3.33 (dd, J=8.6, 13.3 Hz, 1H), 3.10 - 2.97
(m, 1H), 2.78 -
2.67 (m, 2H), 1.86- 1.71 (m, 2H).
fi-NH 1
N 110
c-ç9NH Nj,S'
dab
RP _______________________ H3C=-o a
-s' NaBH(OAc), DCE. 25'C
8 r"-- 31
CI
A mixture solution of Compound 8 (100 mg, 216.73 gmol, 1 eq) and Compound 9
(30 mg,
272.44 gmol, 1.26 eq) in CH3COOH (0.5 mL) and DCE (1 mL) was stirred at 25 C
for 30
min. Then NaBH(OAc)3 (150 mg, 707.74 gmol, 3.27 eq) was added into the mixture
solution, the resultant solution was stirred at 25 C for 1.5 h. LCMS showed
the starting
material was consumed and the desired MS was generated. The mixture was
diluted with
water (50 mL) and extracted with Et0Ac (50 mL*2). The organic layer was washed
with
brine, dried over Na2SO4 and concentrated to dryness, the residue was purified
by Prep-
HPLC (column: Phenomenex Synergi C18 150*25*10 gm; mobile phase: [water
(0.225%
FA)-ACN];B%: 40%-51%, 4 min) to Compound 31 (46 mg, 66.30 gmol, 30.59% yield,
99.7% purity) was obtained as a white solid.
EXAMPLE (III)
Summary of Formula (III)
This invention embodiment relates to compounds having the following formula:
330
CA 3050553 2019-07-25
Formula (III)
R12
w I
(Xlw I aµ
¨GI¨ (kW.
(Xic)k
R1 R2
R3
R5 R4
including
R12
w I
(XII&
N¨(.)(9)9
R1 R2
R3
R5 R4
including
/3¨Th
0 1%
1
X
R.12 (R8)q N
N
R1 R2
R3
R5 R4
including
331
CA 3050553 2019-07-25
%1
N,¨ Rl2x N (R8)q
R2
R1
R3
R5 R4
including
R12,_
(R8)q
R1 R2
R3
R5 R4
including
(L),
147.41µ
NH
(L)õ, 0
o
)c soµ H
(L),
including
332
CA 3050553 2019-07-25
(Om
(L),
N NH
0 0
\OH
oµµ
(L)m (1-)m
or their enantiomers, diastereomers, pharmaceutically-acceptable salts,
solvates, hydrates or
prodrugs thereof, wherein:
Optionally, one or more places have deuterium in place of hydrogen, at an
artificially high
level of deuterium incorporation, in excess of the naturally occuring
abundance;
Optionally, one or more places have fluorine, or other halogen, or methyl, or
alkyl, or
substituted alkyl, in place of hydrogen;
Z is heteroaryl;
g, w and k are independently selected from 0, 1, 2, 3, 4;
.. L is independently at each point of its use hydrogen, alkyl, or substituted
alkyl (non-limiting
example: CF3), or deuterated alkyl (non-limiting example: CD3), or aminoalkyl,
or thioalkyl,
or alkoxy, or halogen, or haloalkyl, or haloalkoxy, or hydroxyalkyl, or any
atom or isotope
permitted by valence (including any accompanying hydrogens by valence e.g.
(non-limiting)
OH, NH2, SH, S1H3, PH2etc.);
Xg, X' and Xk are independently selected from a single bond, 0, S. Se, Nle,
PRv, BRv,
C(Rv)2 or Si(Rv)2, wherein each Rv is independently selected from a
constituent group of L
(defined earlier);
GI is, independently at each point of use, N or CH;
c is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9;
m is independently at each point of use selected from 0, 1, 2, 3, 4, 5, 6, as
valence permits;
R2 is hydrogen, L (defined earlier), hydroxyl (-OH), SH, NH2, methyl, alkoxy,
substituted
alkoxy, haloalkoxy, ether, halogen or ¨0C(0)R14;
R14 is hydrogen, alkyl, haloalkyl, aryl, arylalkyl, cycloalkyl or
(cycloalkyl)alkyl;
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CA 3050553 2019-07-25
R3 and R4 are each independently hydrogen, or L (defined earlier), or CF3, or
NH2, or OH, or
chlorine or other halogen, or alkyl, or substituted alkyl, or deuterated
alkyl, or arylalkyl, or
R3 and R4 taken together with the carbon atom to which they are attached form
a 3- to 7-
membered carbocyclic ring;
R5 is independently at each point of use hydrogen, L (defined earlier), PH2,
OH, SH, alkyl,
substituted alkyl, halogen, nitrile, haloalkyl, aryl, heteroaryl, cycloalkyl
or heterocyclo;
R12 is selected from hydrogen, deuterium, alkyl, aryl, heteroaryl, cycloalkyl,
heterocyclo;
X is alkyl;
Y is a single bond, ¨CH2¨, ¨C(0)¨, ¨0¨, ¨S¨, ¨N(R14) ¨ or (Xf)f where Xf is
selected from a single bond, 0, S, Nle or C(Rv)2, wherein each Itv is
independently selected
from a constituent group of L (defined earlier);
f is 0, 1, 2 or 3;
A is nitrogen (N), or N , or carbon;
E is absent, or alkyl, or substituted alkyl (non-limiting example: CF3), or
deuterated alkyl, or
aminoalkyl, or thioalkyl, or alkoxy or any atom or isotope permitted by
valence (including
any accompanying hydrogens by valence e.g. (non-limiting) OH, NH2, SH, SiH3,
PH2etc.),
for example hydrogen, or deuterium, or fluorine;
R8 is independently selected at each point of use from E (defined earlier),
hydrogen, alkyl,
halogen, carbamyl, carbamylChaalkyl, substituted alkyl or two R8 groups join
to form an
optionally substituted fused phenyl ring;
q is 0, 1, 2, 3 or 4.
R1 is selected from L (defined earlier), hydrogen, deuterium, CN, 502-
piperidine, S02-
piperidine substituted with 0-10 of R5, R9, cyano, halogen, alkyl, substituted
alkyl, alkenyl,
substituted alkenyl, alkylene, substituted alkylene, alkynyl, substituted
alkynyl, alkoxy,
thioalkyl, aminoalkyl, carbamyl, sulfonyl, sulfonamide, cycloalkyl,
(cycloalkyl)alkyl,
hydroxyalkyl, haloalkyl, haloalkoxy, alkoxyalkyl, morpholinylalkyl, aryl,
arylalkyl,
heterocyclo, heteroaryl, (heterocyclo)alkyl, (heteroaryl)alkyl, acyl,
alkoxycarbonyl,
substituted amino;
Most preferably R1 is smaller than 300 Daltons;
R9 is
334
CA 3050553 2019-07-25
0
I I
R7N
"
R6 I I
0
=
,
R6 and R7 are independently hydrogen, L (defined earlier), R1 (provided R1 is
not R9),
alkyl, aryl, heteroaryl, cycloalkyl, heterocyclo, arylalkyl,
(heteroaryl)alkyl, haloalkyl,
hydroxyalkyl, hydroxyalkyl substituted with a carboxylic ester or carboxylic
acid,
alkoxyalkyl, thioalkyl, (cycloalkyl)alkyl, morpholinylalkyl, heterocyclo or
(heterocyclo)alkyl; or R6 and R7 taken together with the nitrogen atom to
which they are
attached form a 5- to 7-membered mono or bicyclic ring including fused rings
such as
1-pyrrolidinyl, 1-piperidinyl, 1-azepinyl, 4-motpholinyl, 4-thiamorpholinyl, 4-
thiamorpholine
dioxide, 1-piperazinyl, 4-alkyl-1-piperazinyl, 4-arylalkyl-1-piperazinyl, 4-
diarylalky1-1-
piperazinyl; or 1-piperazinyl, 1-pyrrolidinyl, 1-piperidinyl or 1-azepinyl
substituted with one
or more L (defined earlier), alkyl, alkoxy, alkylthio, halo, trifluoromethyl,
hydroxy, aryl,
arylalkyl, ¨000R14 or ¨CO-substituted amino;
or R5 and R6 taken together with the atoms to which they are attached form a 5-
to 7-
membered ring optionally substituted with aryl;
Encompassed by this invention are methods of administering a therapeutically
effective
amount of any compound(s) of [P6], or a pharmaceutically-acceptable salt,
solvate, hydrate
or prodrug thereof, optionally in a pharmaceutical composition(s), optionally
in co-therapy
with another anti-cancer treatment(s), to treat/ameliorate/prevent/combat
cancer in a subject.
Especially preferred for this use are compounds of [P6] with 3S, 4R
stereochemistry.
Preferred compounds of Formula (III)
Preferred methods are to use, and preferred compounds are, compounds of
Formula (III),
their enantiomers, diastereomers, pharmaceutically-acceptable salts, solvates,
hydrates or
prodrugs thereof, in which:
Z is triazolyl optionally substituted with one to two R8 or imidazolyl
optionally substituted
with one to two R8 and/or having fused thereto a benzene ring in turn
optionally substituted
with one to two R8;
Y is oxygen;
335
CA 3050553 2019-07-25
R2 is hydroxyl;
R3 and R4 are methyl or chlorine;
R1 is R9;
GI is nitrogen;
R6 and R7 are alkyl; or R6 and R7 taken together with the nitrogen atom to
which they are
attached (GI = N) form a 6-membered ring;
X is alkyl;
R12 is aryl or heterocyclo;
A is N;
E is absent, or deuterium, or hydrogen;
R5 and R8 are hydrogen;
Stereochemistry is 3S, 4R;
Example embodiments of Formula (III)
.. Immediately below, compounds from [7], selected as specific anti-cancer
therapeutics by the
invention of this disclosure. EC50values for FIFO ATP hydrolysis, and FIFO ATP
synthesis, in
NADH-linked and NADPH-linked sub-mitochondrial (SMP) assays respectively. [7]
refers to
these EC50 values as ICsovalues for inhibiting FiFo ATP hydrolase (reverse
mode) and FIF0
ATP synthase (forward mode). However, this in incorrect. Because, as
identified by the
invention of this disclosure, explained herein, although these molecules
inhibit FIFO ATP
hydrolase, their reducing of FIN ATP synthesis is not (predominantly) because
of inhibiting
FiFo ATP synthase, but by uncoupling. The structure on the left is BMS-199264.
It does not
harm ex vivo rat heart at a concentration (10 t.tM [11]) that it exerts anti-
cancer activity
(discovery of this disclosure).
336
CA 3050553 2019-07-25
/-----
i--''¨I--\
Cl N
0 Ny-NH Cl 0 y NH
N
0
' R
OH
0 ON N
=,.'
aCH3 CH3
0 0
CH3 3S,4R CH3
EC50F 1 Fo ATP hydrolase = 0.48 i 0.23 (pM) EC50F1F0 ATP hydrolase = 0.24 t
0.13 (pM)
EC50F1F0 ATP synthesis = 18 t 9.5 (pM) EC50F1F0 ATP synthesis = 3.8 2.1
(pM)
EC50 Ratio = 38 3R,4S
pM doesn't harm ex vivo rat heart EC50 F1F0 ATP hydrolase = 0.48 0.23
(pM)
EC FiFo ATP synthesis = 4 i 0.45 (pM)
For the following example embodiment, with synthesis scheme (as 2 possible
salts shown,
Scheme IHa), the starting material is BMS-199264, which is available
commercially. For
5 example from Sigma-Aldrich, a chemical and reagents vender well known to
those of the art.
Scheme Illa
/....:-.1
41111 õ... ..-NH CI 011) --N
CI ,CH3
--I
N I-13C N"
0õ ,0
R
K2CO3, DMF .õ,-----.N.:-S" R. OH
---...--) s s
CH3 ."....) CH3
0 ,..,,, 0 a...1_1
%A-13 t...1-13
HBF4 HCII
-r CIz--1-.
HN t
Me0H '
CI 0N"
F`F
F\_-õ-F CI N/---1
* Hy.N.t.
...-b N
CH3 0, ,0
N ..---,N;S' 0 R
0\õ0 s
0 CH3
"....--) 0 s CH3 CH3
CH3
Further example embodiment:
337
CA 3050553 2019-07-25
CI *
0 0
PI
C H3
The following example embodiment does not uncouple the proton motive force
(pmf)
because its imidazole group, unlike BMS-199264, for example, does not have a
protonable
element.
H3C-1\14.1"1¨
CI yr\I¨CH3
,0
CH3
CH3
The following example embodiment (logP=3.79, calculated from structure [31])
uncouples
the proton motive force (pmf) less than BMS-199264 (logP=4.35, calculated from
structure
[31]) because its logP is further removed from the logP=-3,2 optimum for
uncoupling [32].
CI
0 0
, HO
.0µ
CI
0
CI
Further example embodiment,
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CA 3050553 2019-07-25
CI
0õ0
OH
0CH3
CH3
Further example embodiment,
CI N \ NH
0, ,0
OH
0 CH3
CH3
BMS-199264, and/or its analogues, can be deuterated by reactions described in
[R1, R2. Ql ,
Q2], which deuterate aromatic and alkyl molecular components. Furthermore,
there is a great
wealth of reactions available to deuterate their aromatic rings, and those
skilled in the art will
know these. For (non-limiting) example, refer [Ex2]. Carbon 1 of BMS-199264, a
chiral
centre, can be deuterated by reactions described in [D, F, El, E2, Exl], which
deuterate a
carbons to tertiary amines, and/or by reactions described in [N], which
deuterate sp3 carbons,
and/or by reactions described in [I, M, G. H], which deuterate a and 0-carbons
to an OH
group. Carbon 6 of BMS-199264, a chiral centre, can be deuterated by reactions
described in
[L, H, G, A, K, M, J1,12, J3, I, F, S], which deuterate the a-carbon to an OH
group, and/or
by reactions described in [D], which deuterate a and 0-carbons to tertiary
amines, and/or by
reactions described in [N], which deuterate sp3 carbons. The scaffold of [P6]
is presented in
its abstract. Deuterated isotopologues of this [P6] scaffold, for (non-
limiting) example
deuterated BMS-199264, are componentry to the present invention as new
compositions of
matter, and in non-limiting embodiments are used singly or in combination,
optionally in co-
therapy with an FDA and/or EMA approved medicine(s) and/or treatment(s), for
example a
licensed cancer treatment, as anti-cancer therapeutics.
EXAMPLE (IV)
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CA 3050553 2019-07-25
Background
Well known to those of the art: amino acids have the following structure,
wherein the R
group is different in different amino acids.
R
H
"N/ <0
H OH
H
Summary of Formula (IV)
This invention embodiment relates to compounds having the following formula:
Formula (IV)
R3
R2 R1 X A
.. j...........G1. i,..,...,,,.s,.0
lik G1 (R1)u
I I R8
01
R6 R7
R4 R5 ( n
)m
Z
including
R2 R1 X A
0
R3 . N7."*Isr"....'......... (R1 )u
I I R8 N
R6 R7
R4 R5 ( n
)m
Z
including
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CA 3050553 2019-07-25
(R8)0.4
NN
\t*
R2 R1 X
R3 N
I R8
gN m
R6 R7
R4 R5
or their enantiomers, diastereomers, pharmaceutically-acceptable salts,
solvates, hydrates or
prodrugs thereof, wherein:
GI is, independently at each point of use, N or CH;
u is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8;
X is selected from 0 or S;
A is selected from hydrogen, deuterium, alkyl, substituted alkyl, cycloalkyl,
heterocycle,
heterocyclo, aryl, heteroaryl, aminoalkyl, thioalkyl, alkoxy and an R group of
a proteogenic
amino acid, or other amino acid synthesized or used by a living system (non-
limiting example
of such a system: a human), which is optionally isotopically enriched, and/or
substituted by
alkyl, substituted alkyl, deuterated alkyl, halogen, cycloalkyl, heterocycle,
heterocyclo, aryl,
heteroaryl, aminoalkyl, thioalkyl, alkoxy, haloalkyl, haloalkoxy, or any atom
or isotope
permitted by valence (including any accompanying hydrogens by valence e.g.
(non-limiting)
OH, NH2, SH, SiH3, PH2etc.);
n and m are 0, 1, or 2;
RI through R5 are independently selected from hydrogen, halogen, NO2, PH2, OH,
SH, CN,
Chsalkyl, substituted Ci_salkyl, C3_8cycloalkyl, aryl, heterocyclo,
heteroaryl, 0R9, SR9,
CORI], CO2R11, CONR9R10 or NR9R10;
R6 and R7 are independently hydrogen, alkyl or substituted alkyl;
R8 is hydrogen, deuterium, Ci_salkyl, substituted Ci_aalkyl, deuterated
Ci_8alkyl, aryl,
heterocyclo, heteroaryl, aminoalkyl, thioalkyl, alkoxy, halogen, haloalkyl,
haloalkoxy, or any
341
CA 3050553 2019-07-25
atom or isotope permitted by valence (including any accompanying hydrogens by
valence
e.g. (non-limiting) OH, NH2, SH, SiH3, PH2 etc.);
Z is hydrogen, alkyl, substituted alkyl, cycloalkyl, aryl, heterocyclo,
heteroaryl, CORii,
CO2R1i, S02R11, S(0)R1 or CONR9R10;
R9 and Rio are independently hydrogen, C1.8alkyl, substituted Ci_salkyl,
C3_10cycloalkyl, aryl,
heterocyclo, heteroaryl, COR13, S02R13 or S(0)R13; and
R11, R12 and R13 are independently hydrogen, Ci_salkyl, substituted Ci_aalkyl,
Cflocycloalkyl,
aryl, heterocyclo or heteroaryl;
wherein each occurrence of R9-R13 is chosen independently.
Preferred compounds of Formula (IV)
Preferred methods are to use, and preferred compounds are, compounds of
Formula (IV),
their enantiomers, diastereomers, pharmaceutically-acceptable salts, solvates,
hydrates or
prodrugs thereof, in which:
R2, R3 and R4 are all hydrogen; and/or
R6 and R7 are both hydrogen; and/or
n and mare both 1; and/or
RI and R5 are both C1-8 alkyl, preferably both Ri and R5 are isopropyl groups.
Other preferred methods use, and preferred compounds are, compounds of Formula
(IV),
their enantiomers, diastereomers, pharmaceutically-acceptable salts, solvates,
hydrates or
prodrugs thereof, in which:
Z is Ci_salkyl, C2_8alkeny1, Ci_shaloalkyl, cycloalkyl, aryl, arylalkyl,
heteroaryl,
heteroarylalkyl ¨CORI 1, ¨0O2R11, ¨SO2R1i, ¨S(0)Ri or ¨CONR9Rio; especially
preferable is benzyl, ¨C(0)2H or ¨C(0)2Ci_salkyl;
R9 is hydrogen;
Rio is Ci_salkyl or C3_10cycloalkyl; aryl or arylalkyl; and
Ri is hydrogen, C _salkyl, C3-locycloalkyl, C3-loheterocycloalkyl, C3.10aryl
or C3-io arylalkyl.
Other preferred methods use, and preferred compounds are, compounds of Formula
(IV),
their enantiomers, diastereomers, pharmaceutically-acceptable salts, solvates,
hydrates or
prodrugs thereof, in which:
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CA 3050553 2019-07-25
A is hydrogen, deuterium, Ci_aalkyl, aminoalkyl, heteroaryl, aryl, or alkyl
substituted with
heterocyclo, aryl, OH, SH, ST'. ¨C(0), H, T3-NT5T6, -T8-C(0)tT9-NT5T6 or T3-
N(T2)T4NT5T6,
TI is alkyl, (hydroxy)alkyl, (alkoxy)alkyl, alkenyl, alkynyl, cycloalkyl,
(cycloalkyl)alkyl,
cycloalkenyl, (cycloalkenyl)alkyl, aryl, (aryl)alkyl, heterocyclo,
(heterocyclo)alkyl,
heteroaryl or (heteroaryl)alkyl;
T2 and T3 are each independently a single bond, -T8-S(0)t-T9-, -18-C(0)-T9-, -
T18-C(S)-T9, _
T8-S-T9-, -T8-0¨C(0)-T9-, -T8-C(0)tT9-, -T8-C(=NT10)-T9- or ¨T8-C(0) ¨C(0)-T9-
;
T5, T6, T7, T8 and T9 are independently hydrogen, alkyl, (hydroxy)alkyl,
(alkoxy)alkyl,
alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, cycloalkenyl,
(cycloalkenyl)alkyl, aryl,
(aryl)alkyl, heterocyclo, (heterocyclo)alkyl, heteroaryl or (heteroaryl)alky,
each group
optionally substituted where valence allows by one to three groups selected
from halo, cyano,
nitro, OH, oxo, ¨SH, alkyl, (hydroxy)alkyl, (alkoxy)alkyl, alkenyl, alkynyl,
cycloalkyl,
(cycloalkyl)alkyl, cycloalkenyl, (cycloalkenyl)alkyl, aryl, (aryl)alkyl,
heterocyclo,
(heterocylco)alkyl, heteroaryl or (heteroaryl)alkyl, ¨STI I, ¨C(0)tH,
¨C(0)tTI I,
¨0¨C(0)T11, T8c(0) )1tN(T12,-.-11,
¨S03H, ¨S(0)JI I, S(0)tN(1-12)T1 I, ¨T'3-NT" T'2, T12, _
T'3¨N(T12)-T4-NTI T22, _T13_N(T11,--.-
) 1 12-
TH and 3-N(T18)-Tm-H; or
T8 and T9 are each independently a single bond, alkylene, alkenylene or
alkynylene;
TI I is alkyl, (hydroxy)alkyl, (alkoxy)alkyl, alkenyl, alkynyl, cycloalkyl,
(cycloalkyl)alkyl,
cycloalkenyl, (cycloalkenyl)alkyl, aryl, (aryl)alkyl, heterocyclo,
(heterocyclo)alkyl,
heteroaryl or (heteroaryl)alkyl;
TI2 is halo, cyano, nitro, OH, oxo, ¨SH, alkyl, (hydroxy)alkyl, (alkoxy)alkyl,
alkenyl,
alkynyl, cycloalkyl, (cycloalkyl)alkyl, cycloalkenyl, (cycloalkenyl)alkyl,
aryl, (aryl)alkyl,
heterocyclo, (heterocyclo)alkyl, heteroaryl or (heteroaryl)alkyl, ¨C(0)tH or
¨S03H;
TI3 and TI4 are each independently a single bond, ¨S(0)t¨, ¨C(0)¨, ¨C(S)--, ¨0-
-,
¨S¨, ¨0¨C(0)--, ¨C(0)t¨, ¨C(=NTI3)-- or ¨C(0)--C(0)--;
wherein each occurrence of T1-T14 is chosen independently; and
t is 1 or 2.
Preferred compounds of the foregoing section are those in which A is hydrogen,
deuterium,
Ci_salkyl, aminoalkyl, hydroxyalkyl, heterocycloalkyl, heteroaryl alkyl, aryl,
arylalkyl, or
alkyl substituted with a group selected from SH, ST4, ¨C(0)H, T6-NT8T9, -III-
C(0)J12-
NT8T9 and T6-N(T5)T7NT8T9.
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CA 3050553 2019-07-25
More preferred are those compounds in which A is hydrogen, deuterium, methyl,
¨
CH2(CH3)2, ¨(CH2)2(CH3)2, ¨CH(CH3)CH2(CH3), ¨(CH2)0H, hydroxyethyl, ¨
(CH2)2SCH3, ¨CH2SH, phenyl, ¨CH2(phenyl), ¨CH2(p-hydroxyphenyl), ¨CH2(indole),
¨(CH2)C(0)NH2, ¨(CH2)2C(0)NH2, ¨(CH2)2C(0)0H, ¨CH2C(0)0H, ¨(CH2)4NF12, ¨
(CH2)3(=NH)CNH2, or ¨CH2(imidazole). Especially preferred A groups are ¨
CH(CH3)CH2(CH3), phenyl, phenyl alkyl or ¨CH2(2-indole).
Alternatively preferred methods use, and preferred compounds are, compounds of
Formula
(IVb), their enantiomers, diastereomers, pharmaceutically-acceptable salts,
solvates, hydrates
or prodrugs thereof, in which:
Formula (IVb)
R1
0 A
0
ii õ----õN
1 R8
gN
R6 R7
R5
Z
including
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CA 3050553 2019-07-25
(R8)04
N\--\
R1
0
=
0
7 11 R8 \
(RN
R5 R6 R7
wherein:
A is selected from hydrogen, deuterium, Ci_salkyl, aminoalkyl, substituted
alkyl, deuterated
alkyl, aryl, heteroaryl, or alkyl substituted with heterocyclo, aryl,
heteroaryl, OH, SH, ST', ¨
C(0)H, T3-NT5T6, -T8-C(0)tT9-NT5T6 or T3-N(12)T4NT5T6;
RI and R5 are independently Ci_salkyl optionally substituted where valence
allows;
R6 and R7 are independently hydrogen or Ci_salkyl;
R8 is hydrogen, halogen, deuterium, C1.8alkyl or substituted Ci_salkyl;
Z is hydrogen, Ci_8alkyl, C2_8alkenyl, Ci_shaloalkyl, cycloalkyl, aryl,
arylalkyl, heteroaryl,
heteroarylalkyl ¨CORI ¨0O21211, ¨SO2RI I, ¨S(0)R11 or ¨CONR9RI ;
R9 is hydrogen,
R'' is Ci_8a1ky1 or C3_10cycloalkyl; aryl or arylalkyl;
RH is hydrogen, Ci_8a1ky1, C3_10cycloalkyl, C3_10heterocycloalkyl, C3_10aryl
or C3_10arylalkyl.
TI is alkyl, (hydroxy)alkyl, (alkoxy)alkyl, alkenyl, alkynyl, cycloalkyl,
(cycloalkyl)alkyl,
cycloalkenyl, (cycloalkenyl)alkyl, aryl, (aryl)alkyl, heterocyclo,
(heterocyclo)alkyl,
heteroaryl or (heteroaryl)alkyl;
T2 and T3 are each independently a single bond, -T8-S(0)t-T9-, -T8-C(0)-T9-,
-T8-S-T9-, -T8 -O ______ C(0)-T9-, -T8-C(0)tT9-, -T8-C(=NTI )-T9 or -T8-C(0)
C(0)-
T9-;
.. T5, T6, T7, T8 and T9 are independently hydrogen, alkyl, (hydroxy)alkyl,
(alkoxy)alkyl,
alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, cycloalkenyl,
(cycloalkenyl)alkyl, aryl,
(aryl)alkyl, heterocyclo, (heterocyclo)alkyl, heteroaryl or (heteroaryl)alky,
each group
345
CA 3050553 2019-07-25
optionally substituted where valence allows by one to three groups selected
from halo, cyano,
nitro, OH, oxo, ¨SH, alkyl, (hydroxy)alkyl, (alkoxy)alkyl, alkenyl, alkynyl,
cycloalkyl,
(cycloalkyl)alkyl, cycloalkenyl, (cycloalkenyl)alkyl, aryl, (aryl)alkyl,
heterocyclo,
(heterocylco)alkyl, heteroaryl or (heteroaryl)alkyl, ¨OTII, ¨ST, ¨C(0)tH,
¨C(0)1T11,
¨0¨C(0)TI I, T8C(0)tN(T12)TI I, ¨S03H, ¨S(0)1TI I, S(0)1N(T12)TI I, -T'3-NT'
'T'2, -T13-
N(T12)-T4-NTIIT22,
-T13-N(Tl I)-T12 -Viand -T13-N(T18)-T14-H; or
T8 and T9 are each independently a single bond, alkylene, alkenylene or
alkynylene;
T" is alkyl, (hydroxy)alkyl, (alkoxy)alkyl, alkenyl, alkynyl, cycloalkyl,
(cycloalkyl)alkyl,
cycloalkenyl, (cycloalkenyl)alkyl, aryl, (aryl)alkyl, heterocyclo,
(heterocyclo)alkyl,
.. heteroaryl or (heteroaryl)alkyl;
T12 is halo, cyano, nitro, OH, oxo, ¨SH, alkyl, (hydroxy)alkyl, (alkoxy)alkyl,
alkenyl,
alkynyl, cycloalkyl, (cycloalkyl)alkyl, cycloalkenyl, (cycloalkenyl)alkyl,
aryl, (aryl)alkyl,
heterocyclo, (heterocylco)alkyl, heteroaryl or (heteroaryl)alkyl, ¨C(0)111 or
¨S03H;
T13 and TI4 are each independently a single bond, ¨S(0)r¨, ¨C(0)--, ¨C(S)--,
¨0¨,
¨S¨, ¨C(0)1¨, ¨C(=NT13)- or ¨C(0)--C(0)¨; and
t is 1 or 2.
More preferred methods/compounds use/are:
A is hydrogen, deuterium, methyl, ¨CH2(CH3)2, --(CH2)2(CH3)2, --
CH(CH3)CH2(CH3), ¨
(CH2)0H, hydroxyethyl, ¨(CH2)2SCH3, ¨CH2SH, phenyl, ¨CH2(phenyl), ¨CH2(p-
hydroxyphenyl), ¨CH2(indole), ¨(CH2)C(0)NH2, ¨(CH2)2C(0)NH2, ¨(CH2)2C(0)0H,
¨CH2C(0)0H, ¨(CH2)4NH2, ¨(CH2)3(=NH)CNH2 or ¨CH2(imidazole).
Especially preferred methods/compounds use/are:
A is ¨CH(CH3)CH2(CH3), phenyl, CH2(phenyl) or ¨CH2(2-indole).
Also, especially preferred methods/compounds use/are:
R8 is hydrogen and the configuration about the carbon marked with the * is S,
provided A is
not H. Also preferred: R8 is deuterium and the configuration about the carbon
marked with
the * is S, provided A is not H or deuterium.
Other preferred methods/compounds use/are:
RI and R5 are both isopropyl; and/or R6R7and R9 are all hydrogen; and/or Z is
CH2(phenyl),
¨C(0)2H or ¨C(0)2C1_8alkyl.
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CA 3050553 2019-07-25
Example embodiments of Formula (IV)
In the following scheme, Scheme X, all reactants are commercially available
e.g. Compound
1 from Matrix Scientific, Columbia, SC, USA, Compound 2 from Oxchem
Corporation, IL,
USA, Compound 4A from Astatech Inc., Bristol PA, USA, Compound 4B from Apollo
Scientific Ltd., Stockport, UK.
Scheme X
CH3 0 N N H
C.) N
CH3 ..- -...
N NaOH3C CH3 N
N=c=0 4. Fi2N av
v. 0 õ.4 , 4B
T
NANO
H3//N. CH3
HO 0 HF H H OH 0 0___/
CH3
2 Fi O
3 ------------------------------------------------------ -1
1 HC "'CH B = ED
CI
i CH2Cl2
(r¨N A H 1
) N N
H3C CH3 N H3C CH3 * r N)
0 v,?0 ...
NAN 0
OH H N 4A NAN-4
H H i_N.,
EDCI
4 f,u r
H3C 'CH3 CH3 112%-.1i 2
H3C CH
I
5A
5B 0 CH3
,
Using Scheme X, above, with different amino acids as the Compound 2 input,
gives different
Compound 5 products. Non-limiting illustration follows: Compound 2b is
available from
Aurora Fine Chemicals LLC, San Diego, USA, Compound 2c and 2d from Sigma-
Aldrich:
N--,---A N--=\
CH3
-,, NH
N¨\ H3C CH3 . NH H3C
0 0
.,.
Nr., NH
.,A 0 NAN 0
H2N Scheme X . 11.1 11 F N 142N Scheme
H H ' N
F (arm A) F (arm B)
HO 0 ,
H3C CH3 HO 0 H3C CH3
2b 2b H3C'NOT
. 0 ;
9
N---=-- \
H3C CH3 ''' N¨CH3 H3C CH3 ,.., N--CH3
N=.-\
-N. N--CH3 I 0 N. N--CH3 0
0 AN NAN 0
H2N Scheme X N H2N Scheme X.
(arm A) -- (arm B)
H H N H H
,
HO 0 HO 0 ---------------- ,--
Fi3C CH3 H3C CH3
2c 2c ..---\
H3C 07
. 0 .
9 9
347
CA 3050553 2019-07-25
¨
HN F HN \ z F
HN H3C CH3 '''' HN H3C CH3 ''
N N
0
arm B)
Scheme ______________________________________ X 411 ,k 0
H2N Scheme )c. (1 N)ls'N H2N N N
H H N
(arm A) (
HO 0 H3C CH HO 0 H3C CH3
2d 2d
H3C o_n
0
In preferred embodiments, the S stereoisomer product of Scheme X is in
enantiomeric excess,
optionally because the Compound 2 input into Scheme X has enantiomeric excess
of S
stereoisomer. To illustrate, with histidine (available from Sigma-Aldrich) as
Compound 2
input,
N=--\ N--.--\
NH -..H
H3C CH3 H3C CH3
N---,A N.=-1
0 0
.tts:H
A 0 4:,,,C1H
A 0
õNH2 Scheme X. 414 N N S NH2 Scheme )c 4110 N NI
H H N
S ' (arm A) S (arm B)
0 OH H3C CH3
2e 2e "
H3C 07
=
0
, ;
Where the amino acid side chain contains an NH group it can optionally be
protected as a
first step, preferably using a protecting group with some degree of
specificity for an amine
over an alcohol group, and for a secondary (NH) over primary (NH2) amine,
wherein greater
specificity is more favoured. Or using an amine protecting group (or chemical
reaction/modification e.g. [non-limiting] refer [242-243]) with some degree of
specificity for
a primary over seconday amine, then protecting the secondary amine with a
different
protecting group, then removing the first protecting group (or reversing the
chemical
reaction/modification of primary amine) with conditions that don't remove the
second
protecting group. Thence a protection group on the NH group of the amino acid
side chain,
and not on the NH2 and OH of the amino acid, is extant. This judicious use of
protecting
groups is inherent to one of the art e.g. refer Greene et al., Protective
Groups in Organic
Synthesis, 3rd Ed., Wiley-Interscience, 1999 (or later edition). Alternatively
one can use an
amino acid starting material with desirable protections incorporated, for
example the starting
material, a protected form of L-histidine (CAS: 274927-61-6), in the scheme
below is
available from multiple suppliers on Labnetwork.com e.g. Astatech Inc.,
Bristol PA, USA
348
CA 3050553 2019-07-25
CH3
Bn 1) NaOH H
CH KII N N---=\
..--,,,,,,
2)CJJ Bn N=C=O H3C CH3 ,,.,. 1.,,,,,N H3C
CH3 NH
N--\\ 0
0
HCI crN
c.
CH3 0 "e
CH3 N)INN4'
t:NF12 Et3N H H 0-CH3 3) H2, Pd/CQr N--11,,N .H H
N
0 9 H3c CH3
H3C CH3
CH3
An illustrating, non-limiting, judicious use of protecting groups is shown
below, wherein
Compound 1 is available from Sigma-Aldrich (609226) and the remaining starting
compounds from suppliers listed on labnetwork.com: Compound 2 (Fluorochem,
Hadfield,
Derbyshire, UK), Compound 5 (Astatech Inc., Bristol PA, USA), Compound 9
(Matrix
Scientific, Columbia, SC, USA) and Compound 12 (Alfa Aesar, Shanghai, China).
0
N=A 0 N=\ N=\
NH CrNH
15"
µr:H 142N-5
2 0¨ N - 0 1 SOCI 10¨
Ji 0
* --.õ _21.
.===
OOH 0 OH 0 CI 9
N=-\ N=-\ N=\
Cc Selectfluor C 1.;
NH l
HN
S-Quinuclidin-3-ol 1
---. -------------.
Cu(OAc)2, Ag2CO3 F-.15.,)-- 7 2) LiOH F b_ 8
6 0¨
0 NH 0 NH 0 OH
AN
N
I ll
N--=\ .1...,,,J N=\
N=\ sCCH CH3
CNH 0
1,5 CH3
0¨
NH2
F 0
fe NCO =N)15N4
15N N2H4o. F
'>=` '.' H HFN
0 ll 2
Cits,1 13
CH LJ
1..
CH3
JJ
11
10 Other example embodiments:
349
CA 3050553 2019-07-25
H3C CH3 0 H3C CH3 0
0 0
.)1. N N S 0 N N 0
H H N H H (N.,
H3C CH3 H3C CH3 1`,.../
0
H3C CH3 143C CH3 111101
0 0
a 0
N N =-= lik --NAN 0
H H N H H rN.,
H3C CH3 1-13C CH3
143C 07
0
Other embodiments:
H3C CH3 0 ...r4;;7,ii H3c CH3 0
¨ )1..
F
O
i-
HN)Nli ,...N.,
0
( / HN HN S N
H3C CH3 H3C CH3
H3C".\041''
0
N ;...,.:.
H3C CH3
--NH H3C CH3
-NH
0 0
NAN74.0 A 0
N N
H H N H H
H3C CH3 H3C CH3
H3C..\oi
cl
Further embodiments:
350
CA 3050553 2019-07-25
H3C CH3 rsd H3C CH3 (rN)
} 9
_
- N N
H H
143C CH3 H3C CH
H3C\ 0 T1
0
() ())
H3C CH3 N) H3C CH3
0 0
NAN 0
H H N H H
H3C CH3 H3C CH3 c.,""
0
Following is Structure IV, from Scheme I, in the "Process of Preparation"
section of [P3],
symbol definitions are as in [P3]. Scheme I in [P3] is a more general form of
Scheme X
above.
1 X
A
R2
OH
IV
This Structure IV can be deuterated, as can final compounds of the scaffold of
[P3] (scaffold
presented in its abstract), at its chiral carbon, and in further embodiments
at further or other
position(s), by reactions described in [A, B, El, E2, F], which can deuterate
the a-carbon to
.. secondary amines. And/or by reactions described in [Ex3], which can
deuterate the a-carbon
to a carbonyl, using pyrrolidine (available from Sigma-Aldrich) as catalyst,
and/or by
reactions described in [Ex4], which deuterate ketones. And/or by reactions
described in [N],
which deuterate sp3 carbons. And/or by reactions described in [RI, R2, Ql,
Q2], which
deuterate aromatic and alkyl molecular components. Deuterated Compound IV
structure(s)
can be inputted into the synthesis Scheme I of [P3], in place of an
undeuterated Compound
IV form compound(s), to make deuterated isotopologue(s) with the scaffold of
[P3], its
scaffold is shown in its abstract. Alternatively, to achieve this aim, a
compound(s) of
Structure III form in Scheme I of [P3] can be deuterated at its chiral carbon,
and in further
351
CA 3050553 2019-07-25
embodiments at further or other position(s), by reactions described in [A, B,
P, El, E2, F],
which deuterate the a-carbon to primary amines. And/or by a methodology used
to deuterate
amino acids, of which many are known to those of the art (non-limiting e.g.
[244-249, B]),
because Structure III (of [P3]) is of the amino acid form. Indeed, deuterated
(and/or other
isotopically enriched e.g. 13C and/or 15N) amino acids can be sourced
commercially, e.g.
(non-limiting) from Sigma-Aldrich or Cambridge Isotope Laboratories Inc., e.g.
look in their
catalogues for options/contact them for further options (e.g. for custom
synthesis), and used
in Scheme I of [P3] to produce isotopically enriched compound embodiments of
the present
invention. For (illustrative, non-limiting) example, Cambridge Isotope
Laboratories Inc. sell
L-histidine enriched (97-99%) for 13C, 15N, 2H at the respective positions of
C, N and H in
histidine (item number: CDNLM-6806-PK). Sigma-aldrich sell this also (item
number:
750158 ALDRICH) and L-tryptophan (749931), L-lysine (749907) and L-
phenylalanine
(749885) enriched for these isotopes also, and L-histidine (791318) and L-
phenylalanine
(CAS: 17942-32-4) enriched for 2H, at multiple positions, only. Cambridge
Isotope
Laboratories sell L-lysine enriched for 2H at multiple positions (DLM-570-PK).
Deuterated
(and other isotopically enriched) compound embodiments of the scaffold of [P3]
(scaffold
presented in its abstract), most preferably deuterated at their chiral carbon
(which in an
embodiment is 13C at enriched, non-natural abundance, e.g. {non-limiting}
>70%13C
incorporation), are componentry to the present invention as new compositions
of matter. And
in non-limiting embodiments, these are used singly or in combination,
optionally in co-
therapy with an FDA and/or EMA approved medicine(s) and/or treatment(s), for
example a
licensed cancer treatment, as anti-cancer and/or anti-aging therapeutics.
To further illustrate by example, without restriction, Compounds 2f (CAS
number: 54793-54-
3), 2g (CAS: 136056-01-4), 2i (L-lysine), 2j (CAS Number: 169524-86-1), 2k
(607665
ALDRICH) are available from Sigma-Aldrich, Compound 2h (CAS: 91037-48-8) from
Cambridge Isotope Laboratories:
H3c cH3 Fi3c cH3
0 0
S
,,,NH2 Scheme )c NH Scheme X.,
N
H H
S D (arm A) S D (arm B)
0 OH H3C CH3 0 OH H3C CH3
2f 2f H3e\DP
0 .
352
CA 3050553 2019-07-25
1101 H3C CH3 *
0 H3C CH3 0
0
A 13c 0 1110 A 13c 0
N N'S'"
13CiNH2 S(cahemAe
H H N 13,,,,NH2 Scheme
----, '-, S (arm B)
===-. H H .õ14...,
0 OH H3C CH3 00H H3C
2g 2g
H3C 0_11
0 ;
N---:---\ N---:--- \
N--=\ I-13C CH3 N, N-CD3
N-=\ H3C CH3 N N-CD3
..c.N-CD3 0
c.,N-CD3 0
Sche 0 ..,kN0
=-. 0
1NH2 me NN X 4110' A S
S (arm A) H H N .5,,,N H Scheme2(arm B)X 10 N
H H N
--- N
0 OH H3C CH3 0"0H H3C CH-4-
2h 2h n,_, ,..õ---\
3k,,, 0T
= 0 ,
5 5
.N, 0 0
NH2 ,....1 .H2
H2NI, H3C CH3 I H2NN HC CI-43
Scheme X. 40' NAN'ACI
Nej:NH2 H H N
s (arm B) ,AH2 ScheITI-12-4.
H H N
(arm A)
c
0 OH H3CXCH3 OOH H3C CH3
21 21 Iv"o_ri
= 0 .
5 5
H2NN H3C CH3 <NH2 H2N <NH2
H3C CH3
0
'IN 0
mu Scheme X, 411 NAN11`e
'113,, ,,..I l'f H H N õõ.., Scheme X 41
XS ' (arm A) 13cs='"' .2 (arm B) H H N
C
0 OH H3C CH3 -)N
0 OH H3C CH3
2j 2j
=
5 0 ;
15,NH2 isc
1,qH2 1,11.i2
13c H3C CH3 13015NNõ. 13C
' --z H3C CH3
l'N 0
LN 0
cheme X NAN0 Scheme X S N N
0
H H N NH.,
_ S
S (arm A) I'-` (arm B) H H .õN...,,
...; ,
0 OH H3C CH3 0 OH H3C CH3 "N.---
2k
S 2k "
H3C 0-7
5 0 ;
5
EXAMPLE (V)
353
CA 3050553 2019-07-25
Formula (V)
11 (R110.4
R2 = 0, S, Se
R3 = 0, C, S, Se, Si R3 0_3
R4 = S, Se L....._hi
R2=R4=R2
R5 = N, C
R6 = R3, X
0-3
R2
R3
(R1)0_3
0
0-3 R3
1 -3 R2
I/
R3 R5
1-3 \
(R1)0_4 R6
I
X
354
CA 3050553 2019-07-25
X
X X R2
R1 = H X I ___________ X
II
R2
AN<C:-
0¨ X
0¨ X f---o o-x
x \_N-x
X
¨X
N¨ XI II
___________________________________________ X X
N=N
X X
X R2 X R2 R2 X
I I E-s-x F-s-x r---s-x 13 1--X I x
I I I I X
X R2 X R2 X X
X = absent, H, Deuterium, OH (hydroxyl), SH (thiol), =0 (keto), CN (cyano),
halogen, CH3 (methyl), methoxy
(OCH3), trifluoromethyl, OCF3, NH2(amino), NOOH (nitro), =N-OH, COOH
(carboxyl), COH (formyl), N=0
(nitroso), 0-N=0 (nitrosooxy), alkyl(C1.4), alkoxy(C1_4), haloalkyl(C1_4),
alkylthio(C14), hydroxyalkyl(C1_4),
aminoalkyl(C1.4), cycloalkyl (C14), haloalkoxy(C1.4), alkenyl(C14),
alkynyl(C14), alkoxycarbonyl(C14),
substituted alkyl(C1_4) (which is an alkyl with between 1 and 4 carbons and
one or more independent
substituents of X)
Molecular permutations of BTB06584. Enumerations of this Markush structure,
and their
pharmaceutically-acceptable salts, solvates, hydrates and prodrugs thereof,
are disclosed as
anti-cancer molecules: the process/method of their use as anti-cancer
molecules is disclosed
by this invention. As valence permits: RI is selected from the options of RI
(independently in
each case of RI), X is selected from the options of X (independently in each
case of X), R2 is
selected from the options of R2 (independently in each case of R2), R3 is
selected from the
options of R3 (independently in each case of R3), R4 is selected from the
options of R4
(independently in each case of R4). In other embodiments one or more phenyl
groups has one
or more of its double bonds replaced with a single bond. In other embodiments,
one or more
phenyl groups is replaced with cyclohexane, each with the same possible
substitutions as the
.. phenyl it replaces. Hydrogen atoms aren't shown in this figure, but in
further embodiments
one or more hydrogen atoms is replaced with deuterium. In further embodiments:
any
possible isotopic substitution at one or more places.
355
CA 3050553 2019-07-25
Example embodiments of Formula (V)
11411111
BTB06584 o=s=o
411
=
- N+
CI 0 0
1411
0=s=0 0=s=0
0 0111 0 0110
(10 0 /10 0
-Nt
CI cy-N--to CI ,0 '0
,
H3C
B1B06584, and/or its analogues, can be deuterated by reactions described in
[RI, R2, Ql,
Q2], which deuterate aromatic and alkyl molecular components. Furthermore,
there is a great
wealth of reactions available to deuterate their aromatic rings, and those
skilled in the art will
know these. For (non-limiting) example, refer [Ex2]. Deuterated isotopologues
of Formula
(V), for (non-limiting) example deuterated BTB06584, are componentry to the
present
invention as new compositions of matter, and in non-limiting embodiments are
used singly or
in combination, optionally in co-therapy with an FDA and/or EMA approved
medicine(s)
and/or treatment(s), for example a licensed cancer treatment, as anti-cancer
therapeutics.
EXAMPLE (VI)
Encompassed by this embodiment are methods of treating a subject suffering
from cancer by
administering an effective amount of at least one compound of Formula (VI) or
a
356
CA 3050553 2019-07-25
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof, or a
pharmaceutical
composition(s) comprising one or compounds of Formula (VI).
Summary of Formula (VI)
This invention embodiment relates to compounds having the following formula:
Formula (VI)
RB
b( M I LA
X
Q
xa(4)
Qr = )xa
.0)%...\\
RA1 cr N
b(
QB QB
b( LY-RA2
X
including
RB
N N
NH
RA2
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
each QA is independently selected from N and CH;
each QB is independently selected from 0, S, Se, NH, CH2, NRw, PRw, BRw,
C(Rw)2 and
Si(Rw)2;
357
CA 3050553 2019-07-25
each M is independently selected from 0, S. Se, NH, CH2, NRw, PRw, BRw, C(R)2
and
Si(Rw)2;
each Rw is independently selected from hydrogen, deuterium, halogen (e.g. F),
alkyl, or
substituted alkyl (non-limiting examples: CF3, CC13), or deuterated alkyl (non-
limiting
example: CD3), or aminoalkyl, or thioalkyl, or alkoxy, or halogen, or
haloalkyl, or
haloalkoxy;
xa is independently at each point of use selected from 1, 2, 3, 4, or 5; xb is
independently at
each point of use selected from 0, 1, 2, 3, 4, or 5; LA represents 0-5
optional substituents on
the ring independently selected from alkyl, substituted alkyl, deuterated
alkyl, aminoalkyl,
thioalkyl, alkoxy, halogen, haloalkyl, haloalkoxy, or any atom or isotope
permitted by
valence (including any accompanying hydrogens by valence e.g. (non-limiting)
OH, NH2,
SH, SiH3, PH2 etc.);
RAI and RA2 are each independently selected from the groups
R C
RD " _________________________________________________ RE
and
wherein Rc and RD are each independently selected from hydrogen, deuterium,
halogen and
alkyl, and wherein RE is hydrogen, deuterium, halogen or alkyl;
le is selected from RBI, hydrogen and deuterium;
wherein RBI is selected from phenyl, benzyl, heteroaryl, pyridyl, pyrimidyl
and
pyrazinyl optionally substituted with one or more substituents RB2;
wherein each RB2 is independently selected from halogen, alkyl, substituted
alkyl,
deuterated alkyl, alkoxy, nitro, amino, methoxy, haloalkyl, polyhalogen alkyl,
aminoalkyl,
thioalkyl, alkoxy, haloalkoxy, and any atom or isotope permitted by valence
(including any
accompanying hydrogens by valence e.g. (non-limiting) OH, NH2, SH, SiH3, PH2
etc.);
or le is a phenylalkyl of the formula:
358
CA 3050553 2019-07-25
. G _______________________________________ (RG)q
(RF)q
wherein RF and RG are hydrogen or alkyl, G is a carbon-carbon double bond or a
carbon-
carbon single bond, n is 0 or 1 and q is 0 or 1 provided that where q is 0, G
is a carbon-carbon
double bond and where q is 1, G is a carbon-carbon single bond,
or RH is a diphenylalkyl of the formula
R H 1
_
it P
RH2
wherein RH' and RH2 each independently represent 1-5 optional substituents on
each ring, and
wherein each RHI and RH2, when present, is independently selected at each
point of use from
hydrogen, deuterium, halogen, alkyl, substituted alkyl, deuterated alkyl,
alkoxy, nitro, amino,
.. methoxy, haloalkyl, polyhalogen alkyl, aminoalkyl, thioalkyl, alkoxy,
haloalkoxy, and any
atom or isotope permitted by valence (including any accompanying hydrogens by
valence
e.g. (non-limiting) OH, NH2, SH, SiH3, PH2 etc.), and p is 0, 1,2 or 3;
or RH is the group
359
CA 3050553 2019-07-25
RJ
.
(GTA\ IL
,QH
(Gu)u"
0
RK
including
Rj
e
Q--1
.
RK
including
360
CA 3050553 2019-07-25
Fe
RK
Wherein GT and Gu are each independently selected from a single bond, 0, S, Ne
or
C(Rv)2, wherein each le is independently selected from hydrogen, deuterium,
alkyl, or
substituted alkyl (non-limiting examples: CF3, CC13), or deuterated alkyl (non-
limiting
example: CD3), or aminoalkyl, or thioalkyl, or alkoxy, or halogen (e.g. F), or
haloalkyl, or
haloalkoxy;
u and t are each independently selected from 0, 1, 2, 3 and 4;
Q is C, CH or N, Wand RK each independently represent 1-5 optional
substituents on
each ring, and wherein each IV and each RK, when present, is independently
selected from
deuterium, halogen, alkyl, substituted alkyl, deuterated alkyl, alkoxy,
haloalkoxy, methoxy,
nitro, amino, aminoalkyl, thioalkyl, haloalkyl, polyhalogen alkyl, and any
atom or isotope
permitted by valence (including any accompanying hydrogens by valence e.g.
(non-limiting)
OH, NH2, SH, Sift, PH2 etc.);
L is absent (when Q is N), alkyl, or substituted alkyl, or deuterated alkyl,
or
aminoalkyl, or thioalkyl, or alkoxy, or halogen, or haloalkyl, or haloalkoxy,
or hydroxyalkyl,
or any atom or isotope permitted by valence (including any accompanying
hydrogens by
valence e.g. (non-limiting) OH, NH2, SH, SiH3, PH2 etc.).
In some embodiments, when one or both of R3 and RK is alkoxy, this alkoxy
group may be
methoxy.
It is to be understood that in the compounds of general Formula (VI), wherein
RA' and/or RA2
are alkenyl moieties having different substituents at the position Rc and RD,
that compound
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CA 3050553 2019-07-25
may exist in cis or trans isomeric forms both of which are considered to be
within the scope
of the present invention. All isotopic, including radionuclide, forms of
Formula (VI) are
within the scope of the present invention.
Preferred embodiments of Formula (VI)
For Formula (VI), the symbols Rc and le, as defined in subgroups RAI and RA2,
may be
hydrogen, halogen (suitably fluorine, chlorine or bromine), alkyl, suitably
"lower alkyl"
(herein now defined) having from 1 to 5 carbon atoms, such as methyl, ethyl,
propyl,
isopropyl, butyl, isobutyl, tert butyl, pentyl and the like, most preferably
methyl;
and the moiety RE may be hydrogen, or lower alkyl having from 1 to 5 carbon
atoms such as
methyl, ethyl, propyl, butyl, or pentyl, most suitably methyl.
The subgroup RB may be hydrogen; phenyl; or substituted phenyl. The
substituted phenyl
group may include one or more of the preferred substituents in any of the
available positions
for substitution, however, mono substitution in the 4-position of the phenyl
nucleus is
.. especially preferred. Suitable substituents for the phenyl nucleus include
halogen, preferably
fluorine, chlorine or bromine; lower alkyl, lower alkoxy, and poly halogen
lower alkyl (i.e.
substituted alkyl) wherein the alkyl moiety contains from 1 to 5 carbon atoms,
especially
preferred however are methyl, methoxy, and trifluoromethyl; and nitro and
amino.
Where the subgroup RB represents substituted pyridyl, substituted pyrimidyl,
or substituted
pyrazinyl, the substituting group may be located on one or more of the
available carbon
atoms in the nucleus, and may be the same or different. Preferred among the
substituting
groups are lower alkyl or lower alkoxy having from 1 to 5 carbon atoms such as
methyl,
ethyl, butyl or penty; or methoxy, propoxy, butoxy or pentoxy.
Where the moiety le represents substituted benzyl, the benzyl moiety may be
substituted in
one or more of the available positions on the phenyl nucleus thereof. Among
the preferred
substituents are halogen (suitably fluorine, chlorine or bromine), lower
alkoxy having from 1
to 5 carbon atoms, especially preferred is methoxy and most preferred being di-
and tri-
methoxy; or alkylenedioxy suitably lower alkylenedioxy such as methylenedioxy,
ethylenedioxy, propylenedioxy and the like, most suitably, the alkylenedioxy
moiety is
attached across the 3- and 4-positions of the phenyl nucleus, although the
bridging of other
carbon atoms in the phenyl nucleus is to be considered within the scope of the
present
invention.
The moieties RF and RG may be hydrogen, or lower alkyl of 1 to 5 carbon atoms,
most
preferred however being methyl.
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CA 3050553 2019-07-25
The groups RBI and RIL2 may be independently hydrogen, or halogen suitably
fluorine,
chlorine or bromine.
Preferred embodiments of Formula (VI) include wherein Rc and RD are methyl, RE
is methyl
and RB is selected from chlorophenyl, methylphenyl, methoxyphenyl,
trifluorophenyl,
chlorophenyl, dimethoxybenzyl, trimethoxybenzyl, methylenedioxybenzyl and
ethylenedioxybenzyl.
In some embodiments RB is the group
411
RK
In some embodiments, RB is the group
RL
RM
wherein RL and Rm are each independently selected from halogen, alkyl, alkoxy,
nitro, amino
= and polyhalogen alkyl.
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CA 3050553 2019-07-25
Synthesis of structures of Formula (VI)
Synthesis routes for example embodiments of Formula (VI) are in [P7], which is
herein
incorporated in entirety by reference. One or more chemical
enumerations/structures from
[P7], in use as an anti-cancer therapeutic, is componentry to the present
invention. Indeed,
encompassed by this embodiment are methods of treating a subject suffering
from cancer by
administering an effective amount of at least one compound from [P7] or a
pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof, or a pharmaceutical
composition(s)
comprising one or compounds from [P7].
Example embodiments of Formula (VI)
Invention embodiments include compounds of Formula (VI), whether a decoupler
or not, and
any compound(s) of any formula, which has decoupling activity (changes FIFO
ATP synthase
stoichiometry), in use for anti-cancer therapy.
Decoupler drugs as anti-cancer medicines
Enumerations of Formulas I-V exert anti-cancer activity by inhibiting, and so
reducing, FiFo
ATP hydrolysis. The present embodiment also exerts anti-cancer activity by
reducing FIE.
ATP hydrolysis. However, not by inhibition of FIR) ATP hydrolysis, but by
making FiFo
ATP hydrolysis more efficient! Such that less ATP is hydrolysed per unit
proton motive force
(pmf) generated i.e. FIFO ATP hydrolysis is reduced. The shared feature of
these
embodiments is that cancer function is impaired, and cancer danger reduced, by
reducing
FiFo ATP hydrolysis in cancer cells. The present embodiment relates to and
discloses the
method/use of a "decoupler" drug(s) as an anti-cancer therapeutic e.g. (non-
limiting)
almitrine, which is a compound of Formula (VI). Disclosed experimental data
shows that
almitrine exerts anti-cancer activity (Figure 7).
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A A
I A A I
A., õGõ, I I G,, A
G G
II I A II
A¨Gõ I õ..Gõ
G
A A
.¨A
A¨G
G:ZA
A A i5N
13c
N N A A E E N
H2C NH Pk
õ/ G IA E N N NH
I
A A
Almitnne GA11
G = 12C,13C (enriched, >NA) I I
CH2 A¨G¨A CH2
E 14N, 15N (enriched, >NA)
A = 1H, 2H (enriched, >NA), F, 32CI (enriched, >NA),
halogen, alkyl, substituted alkyl, CF3, deuterated alkyl
NA = natural abundance
Decoupler drugs modify the Fr/ATP stoichiometry of ATP synthase, so modifying
the
ATP/0 ratio, without significantly changing Tim [250-253]. Such drugs
symmetrically
modify the forward and reverse modes of ATP synthase: they make the forward
mode less
efficient (less ATP synthesized per protons passed) and the reverse mode more
efficient
(more protons pumped per ATP hydrolysed). In other words, they reduce the
(fractional)
ATP yield/cost of a proton passing through ATP synthase energetically
"downhill"/"uphill",
in relation to the direction of the proton motive force (pmf). In isolated
mitochondria, the
almitrine conferred decrease in FiFo ATP synthesis and hydrolysis is maximal
at 40%, no
matter how great the almitrine concentration [251]. Almitrine can double the
stoichiometry:
double the number of protons required/pumped for/by ATP synthesis/hydrolysis.
Almitrine
reduces the amount of ATP that FiFo ATP synthase synthesizes, but it also
reduces the
amount of ATP that FIFO ATP synthase hydrolyses, which is normally, without
almitrine,
significant. Indeed, this disclosure discloses a new fundamental biological
discovery, with
supporting in vivo experimental data (Figure 23): FiFo ATP hydrolysis isn't a
bug but a
feature, substantial and essential to heat generation and homeothermy.
Almitrine reduces
ATP synthesis and hydrolysis and these aspects largely offset and
intracellular [ATP] remains
within survivable limits. Almitrine reduces inefficiency (heat generation) but
simultaneously
increases inefficiency (heat generation). Making the forward mode of ATP
synthase less
efficient might be bad for normal cells, as it might be for cancer cells that
use OXPHOS, but
this decoupler action disproportionally affects cells with a higher
respiratory rate [251],
which could disproportionally affect cancer cells. Although decouplers make
the reverse
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mode of ATP synthase more efficient, this action still exerts anti-cancer
activity on cancers
using ATP synthase in reverse. Because these cancers use this mode for its
inefficiency: to
consume ATP, to permit high glycolytic and PPP flux, to enable high [NADPH]
and low
[ROS]. So, slowing/reducing this FIFO ATP hydrolysis exerts anti-cancer
activity. The same
cancer can be disrupted by both almitrine actions, upon the forward and
reverse modes of
ATP synthase, at different stages of the cell cycle.
In normal cells, almitrine decreases ATP synthesis, but decreases ATP
hydrolysis also, and so
[ATP] is maintained. In cancer cells residing in hypoxia, thence forced to
survive with a
lower OXPHOS rate, which already rely upon high IFI expression (many cancers
overexpress
[23-24]) to block ATP hydrolysis and buoy [ATP], almitrine conferred decrease
in ATP
synthesis strikes them disproportionally.
Comparing Figure 1 and 7A, one observes that in standardized NCI one-dose
testing [16-17],
almitrine dismesylate (10 M) exerts greater anti-cancer activity than
carboplatin (10 M),
which is an FDA approved chemotherapeutic, one of the most used chemotherapies
today,
and is on the World Health Organisation (WHO) list of essential medicines.
Furthermore,
almitrine dismesylate is less toxic to normal cells than carboplatin. In mice,
intraperitoneal
injection (IP) LD5o (dosage at which 50% of test mice die) = 118 mg/kg for
carboplatin [254]
and = 370 mg/kg for almitrine dismesylate [255].
In humans, 200 mg per day of oral almitrine dimesylate has been trialled for
sleep apnea,
which is a drive to snoring [256]. In humans, oral almitrine dimesylate has
been used for
decades, totalling millions of patient months of almitrine administration, for
chronic
obstructive pulmonary disease (COPD), often at 200 mg oral almitrine
dimesylate per day
[257, 258], wherein 400 mg per day has been shown safe in humans [259]. In
humans, a
single 200 mg oral almitrine dimesylate dose (>85% orally bioavailable [260])
renders a
mean Cmax plasma concentration of 286 ng/ml = 0.6 M, AUC(0-72 hours) = 3943.2
(ng*h)/ml,
AUC(0.03) = 6248 (ng*h)/m1 [258]. Using [Css= AUC(0.00/dosing interval] [261],
wherein Css
is drug plasma concentration at steady state, wherein 200 mg is administered
per day (thence
dosing interval = 24 hours), Cs, = 6248/24 = 260.3 ng/ml = 0.55 M. Because
almitrine
volume of distribution (VD) = 17 1/kg [258], and human volume = 1/kg [262],
corresponding almitrine tissue concentration (assuming uniform) at steady
state = (0.55*17)
= --10 M, which is an almitrine concentration that exerts anti-cancer
activity in NCI testing
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(Figure 7A). Almitrine has a long half-life in the body, so daily doses
fractionally compound
and drive higher and higher almitrine plasma concentrations and, indeed, in
humans, oral 50
mg almitrine dimesylate, twice per day, for 1 year produced an average steady
state almitrine
plasma concentration on the 3651h day of 432.8 ng/ml (daily peak plasma
concentration in
some patients exceeded 1000 ng/ml); almitrine plasma levels didn't stop
increasing and
stabilize until a point between day 90 and day 180 [263]. Extrapolating from
[263], oral 200
mg per day almitrine dimesylate (a dosage used in human therapy [257, 258])
produces
average steady state almitrine plasma concentration on the 365th day of 865.6
ng/ml = 1.81
M, (1.81*17) = 30.8 M tissue concentration of almitrine, which is 3 times
greater than an
almitrine concentration, 10 M, that exerts significant anti-cancer activity
in NCI testing
(Figure 7A). A different human study [264] recorded a higher almitrine volume
of
distribution (39.23 1/kg), which translates to a greater tissue concentration
of almitrine:
(1.81*39.23) = 71 M. In clinical use for COPD, a plasma concentration of 300
ng/ml
almitrine is the directive [257], which corresponds to an anti-cancer tissue
concentration (-10
M to >10 M) of almitrine. Non-limiting anti-cancer embodiments of this
invention are
almitrine dosages/formulations/compositions/salts/patterns of administration
(e.g. sequential
administration scheme) already used in humans (for example, as reported in the
literature).
Furthermore, higher or lower almitrine doses for anti-cancer therapy,
optionally administered
intravenously and/or with layoff periods (no drug administered), are further
embodiments of
.. this invention.
Five minutes after i.v. bolus administration of radioactively labelled "C-
almitrine to
anesthetized rats, almitrine in plasma is 0.69% dose/g and almitrine has
disproportionally
distributed to the lungs [12.26% dose/g = 17.77 times the plasma
concentration, this disparity
rises to 75.3 times at 15 minutes because plasma falls faster than lung
concentration] (bronchi
[10.62% dose/g]), adrenal glands [10.66% dose/g], liver [2.55% dose/g]
(although a
significant amount of radioactivity detected in liver isn't because of intact
almitrine but
because of one or more of its metabolite [s] ; if accumulated by the liver,
almitrine is
significantly broken down), kidney [1.58% dose/g], heart [2.73% dose/g],
carotid body
.. [2.66% dose/g] and superior cervical ganglion (SCG) [0.26% dose/g, but
increases to 1.81%
dose/g by 2 hours] [266]. The same experiment was performed without the rats
being
anesthetized, with the Pt recording time point two hours after i.v. bolus,
wherein this was the
last recording time point for the prior experiment with anesthetized rats, and
so at this shared
time point, differences in almitrine tissue distribution between anesthetized
and
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unanesthetized rats can be identified. There was no notable difference for any
of the
aforementioned tissues, except for (anesthetized vs. unanesthetized) the
caratoid body (1.18
vs. 0.58 % dose/g), SCG (1.81 vs. 0.22 % dose/g), which both had less
almitrine, and the
liver (0.88 vs. 2.51 % dose/g), which had more. Relating the tissue types that
disproportionally accumulate almitrine in rats to the case of humans
administered with
almitrine: the almitrine concentration in one or more of these human tissues,
most especially
the lung (especially the bronchi), might exceed the human tissue calculation
disclosed earlier,
which treated the entirety of the tissues as a single uniform compartment (in
the case of
lungs, it at least matches it on the basis of the rat data, wherein lung has
17.77 times greater
than plasma concentration of almitrine, 5 minutes after i.v. bolus of
almitrine, which is close
to the multiplicative constant, 17, that I used in my earlier calculation).
Therefore, in
invention embodiments, almitrine or a pharmaceutically-acceptable salt,
solvate, hydrate or
prodrug thereof is especially suited, without limitation, to treating a cancer
in one or more of
these regions in a subject. For example,
treating/ameliorating/preventing/combating a lung
cancer(s) such as, without limitation, small-cell lung carcinoma (SCLC), non-
small-cell lung
carcinoma (NSCLC, including, without limitation, adenocarcinoma of the lung,
bronchioloalveolar lung cancer, bronchioloalveolar carcinoma, squamous-cell
carcinoma of
the lung, large-cell lung carcinoma, pleomorphic, carcinoid tumor, salivary
gland-like
carcinoma, unclassified carcinoma, rhabdoid carcinoma, sarcomatoid carcinoma,
adenosquamous carcinoma, papillary adenocarcinoma, giant-cell carcinoma, an
admix of
NSCLC types, "not otherwise specified" type), combined small-cell lung
carcinoma (c-
SCLC), pancoast tumors, carcinoid tumors, bronchial gland carcinomas,
sarcomatoid
carcinomas and non-carcinomas (such as, without limitation, sarcoma, lymphoma,
immature
teratoma, melanoma), a cancer(s) listed in [267], cancer(s) in a
bronchus/bronchi, bronchial
adenoma. Almitrine or a pharmaceutically-acceptable salt, solvate, hydrate or
prodrug thereof
is particularly suited to treat/ameliorate/prevent/combat Non-Small Cell Lung
Cancer
(NSCLC) because in vitro data shows almitrine dimesylate to exert anti-cancer
activity
against NSCLC cell lines (Figure 7, 31.6% mean growth inhibition for NSCLC
cell lines
when administered at 10 uM). At 10 M, almitrine dimesylate exerts greater
anti-cancer
activity than carboplatin (Figure 1, 6.3% mean growth inhibition for NSCLC
cell lines when
administered at 10 p,M), a drug in present clinical use for NSCLC. At 100 M,
the maximum
growth inhibition that almitrine dimesylate exerts against an NSCLC cell line,
= 76.46%
growth inhibition (Figure 7B). Moreover, the way almitrine exerts anti-cancer
activity,
disclosed elsewhere herein, it is suited to treat chemoresistant and/or
radioresistant lung
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cancer(s) because it undermines a mechanism by which these cancers have radio-
and/or
chemo- resistance, as disclosed elsewhere herein. Futhermore, almitrine or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof is
especially suited,
without limitation, to treat/ameliorate/prevent/combat a cancer(s) of the
caratoid body, such
.. as carotid paraganglioma (carotid body tumor), and/or a liver cancer(s)
such as hepatocellular
carcinoma, cholangiocarcinoma, hepatoblastoma and/or a kidney cancer(s) such
as renal cell
carcinoma (RCC), renal oncocytoma, transitional cell carcinoma (TCC), squamous
cell
carcinoma, juxtaglomerular cell tumor (reninoma), angiomyolipoma, Bellini duct
carcinoma,
clear-cell sarcoma of the kidney, mesoblastic nephroma, metanephric adenoma,
cystic
nephroma, Wilms' tumor, mixed epithelial stromal tumor and/or a heart
cancer(s) (primary
and/or secondary) such as papillary fibroelastoma, rhabdomyoma, angiosarcoma,
teratoma,
cystic tumour of the atrioventricular nodal region and/or a cancer of the
adrenal gland such as
adrenocortical adenoma, adrenocortical carcinoma, neuroblastoma,
pheochromocytoma, and
paraganglioma.
An embodiment of the invention is administering almitrine or a
pharmaceutically-acceptable
salt, solvate, hydrate or prodrug thereof to a subject with cancer, optionally
lung cancer
and/or mesothelioma of the lung, wherein this subject has difficulty breathing
and/or has low
blood p02 and/or high blood pCO2 and/or Chronic Obstructive Pulmonary Disease
(COPD)
and/or Acute Respiratory Distress Syndrome and/or is undergoing (or will
undergo or has
undergone) surgery including anti-cancer surgery including surgery to remove
lung cancer
including surgery to remove part or the entirety of a lung (pneumonectomy)
including open-
chest thoracic surgery (thoracotomy) including open-chest one lung
ventillation, optionally in
co-administration with (e.g. inhalation of) Nitric Oxide, NO (illustratively,
not restrictively,
NO at 10 parts per million [p.p.m]) and/or hyperbaric 02 therapy. Almitrine or
a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof is
especially suited to
treating lung cancer because it exerts anti-cancer activity, thence
fundamentally treating the
symptoms of lung cancer, and concurrently directly treats the shortness of
breath/beathlessness symptom of lung cancer. Without limitation to any
mechanism(s), the
former action is by almitrine effect upon ATP synthase in cancer cells, the
latter action is by
almitrine effect upon BK potassium channels in the caratoid bodies, wherein
this latter action
also exerts anti-cancer activity, adding/potentiating with the first, because
it increases tissue
p02 and thence ROS production. Elegantly, almitrine both increases ROS (by
increasing
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blood and tissue p02) and decreases ROS mitigation, especially in cancer cells
(by slowing
FIR) ATP hydrolysis).
Almitrine acts upon BK potassium channels in chemoreceptors, within the
caratoid bodies,
and acts as a respiratory stimulant, which increases blood and tissue
oxygenation, decreasing
their [CO2] [268, 269]. This respiratory stimulation should exert an
additional anti-cancer
effect in vivo because increasing [02] in blood and tissues increases their
[ROS], especially in
combination with ROS inducing [chemo/radio] therapies (permitting their use at
lower doses,
reducing their side effects). This synergises with the almitrine conferred
reduction in Fi Fo
ATP hydrolysis in cancer cells, which corrupts the system cancers use to
maintain low
intracellular [ROS] at key stage(s) of the cell cycle, which is paramount to
their "limitless
replicative potential" (Hallmark of cancer [26]) and thence danger. Almitrine
will be
especially valuable against cancers (e.g. lung, breast) that can disrupt
breathing and/or reduce
02 delivery to tissues. Embodiments of this invention are to use almitrine, or
any other
drug(s) that modifies ATP synthase stoichiometry (a decoupler), as an anti-
cancer medicine,
optionally in co-therapy with one or more FDA and/or EMA approved drug(s),
e.g. a cancer
drug(s), and/or in co-therapy with any other compound(s) embodiments of the
present
invention e.g. a compound(s) of Formula (I-V) herein. Almitrine dimesylate is
also known as
almitrine bismesylate or almitrine dimethanesulfonate. All pharmaceutical
salts of almitrine
are contemplated as anti-cancer therapeutics, as is almitrine in complex with
another drug(s)
e.g. almitrine-raubasine.
When used chronically, almitrine dimesylate can have side effects [257]. A 30
year national
pharmacovigilance survey in France, representing several million patient
months of oral
.. almitrine dimesylate treatment [265], showed that upon multi-year use (mean
onset of
adverse reactions = 11 months), some patients receiving oral almitrine
dimesylate for COPD
exhibited weight loss (795 cases) and peripheral neuropathy (2,304 cases)
[257]. Although
these side effects only presented in a minority of patients and only in a sub-
minority (<10%)
of these were they categorised as serious [257]. In most cases, these side-
effects were
.. reversible. Almitrine has never been submitted for FDA approval (doxapram
is a drug
alternative to almitrine, also a respiratory stimulant, which is FDA approved,
wherein
doxapram was found first, which may, to speculate, have been a factor in
dissuading an
{expensive} FDA application for almitrine). Oral almitrine dimesylate has now
been
withdrawn from use in France, Portugal and Poland, where it was previously
approved to
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treat chronic obstructive pulmonary disease (COPD). This withdrawal was
because of the
aforementioned two side effects and because "available efficacy data,
including data which
became available since the initial marketing authorisation, showed only very
limited clinical
efficacy of oral almitrine in its approved indications" [257]. Although oral
almitrine does
increase arterial p02, this does not translate to significant clinical benefit
for COPD sufferers
[257]. Injectable almitrine dimesylate was reviewed separately, in a review of
its use to treat
its own indications, distinct from the review in 2013 of oral almitrine
dimesylate for COPD
[257]. It was reviewed in France by the Haute Autorite De Sante in 2013 [270].
After this
review, injected almitrine dimesylate is still used in France ("Special
status, List II, medicinal
product for hospital use only") wherein "the [review] Committee recommends the
continued
inclusion of VECTARION [brand name for almitrine dimesylate in France, where
it is made
by Servier], lyophilisate and solution for preparation for injection, in the
list of medicines
approved for hospital use in hypoxaemia and hypercapnia associated with
alveolar
hypoventilation when weaning patients off artificial ventilatory support and
at the dosages
indicated in the Marketing Authorisation". From this report, "the adverse
effects observed
with injectable almitrine are paradoxical breathing difficulties and possible
digestive
disturbances such as nausea, heartburn and bloating...these effects do not
routinely entail the
discontinuation of treatment". Up to 25/11/2011, the number of treatment days
with
injectable almitrine in France was estimated at 9,453 patient-months and from
26/11/2006 to
25/11/2011 (5 years) there were only two pharmacovigilance reports: "one case
of a non-
serious increase in lactic acidaemia with a plausible causal link to VECTARION
for
injection, and one case of Stevens-Johnson syndrome in a polymedicated female
patient
under treatment with injectable VECTARION". Injectable almitrine is also
available/used in
some other jurisdictions, e.g. for "specialist and hospital prescribing" by
the British National
Health Service (NHS) [271]. Why does oral almitrine dimesylate for COPD
treatment
produce side-effects in a small number of cases [265], whereas injected
almitrine dimesylate
produces less side-effects [270]? Probably because oral almitrine dimesylate
for COPD
treatment tends to be used chronically, over months to years, whereas
injectable almitrine
dimesylate tends to be used acutely, over day(s). And it is chronic almitrine
use (mean onset
of adverse reactions = 11 months [265]) that can cause, typically non-serious
and reversible,
side effects in a minority of patients. Acute almitrine administration is even
safer. Thence
there is a therapeutic window for using almitrine, very safely, for acute anti-
cancer treatment.
Moreover, I argue later in this disclosure that the side-effects observed with
chronic almitrine
use for COPD are very tied to the COPD pathology itself, and won't be seen so
prevelantly
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with chronic almitrine use for anti-cancer treatment. But anyhow, chronic
administration
won't be needed/chosen in many cases. Significant cancer treatment will come
from using
almitrine acutely or non-continuously, with breaks in almitrine treatment.
Indeed, present
radio- and chemo- therapies are extremely toxic and so can only be
administered infrequently
(e.g. by FDA label: a cisplatin dose [100 mg/m2] should not be administered
more frequently
than every 3 to 4 weeks, e.g. a carboplatin dose [360 mg/m2] should not be
administered
more frequently than every 4 weeks; this instruction is not because these
drugs stay in the
body for an especially long time, they don't, but because they are just so
toxic that the body
needs time to recover inbetween doses) and typically for a capped number of
times, or
regularly (e.g. daily) for a short period, or for cycles of a short period of
administration
alternating with periods of non-administration inbetween them. Alm itrine or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof can be
administered
exactly or approximately matching, or some function of, the pattern of
administration of one
or more of chemo- and/or radio- therapy given to the subject, wherein
almitrine will
synergize with their anti-cancer effects by a mechanism that will be disclosed
later in this
disclosure. To illustrate, an invention embodiment is to administer almitrine
or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof (oral
and/or injected)
every day that radiotherapy is administered, wherein illustrative (not
restictive) courses of
radiotherapy are to administer it every week day (one or more times) for a
period that can
vary between 3 to 9 weeks, or 3 times per day for 12 days (continuous
hyperfractionated
accelerated radiotherapy, CHART) or (especially with stereotactic
radiotherapy) 3 to 8 times
over 2 to 3 weeks, or accelerated fractionation, or hyperfractionation, or
hypofractionation
radiotherapy administration schemes. In further embodiments, almitrine or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof is also
administered
(oral and/or injected) at weekends during a weekday radiotherapy course,
and/or during
pauses in radiotherapy treatment and/or is administered (oral and/or injected)
for a further
period of administration flanking one or both sides of a radiotherapy
treatment period. In
some embodiments, almitrine or a pharmaceutically-acceptable salt, solvate,
hydrate or
prodrug thereof is administered during a period before and/or during and/or
after anti-cancer
surgery.
Oral almitrine dimesylate (100-200 mg) was administered to human COPD patients
daily for
a year [266]. In a minority, peripheral neuropathy started to manifest (5
times higher
incidence in almitrine than control group) by 7 months on average. Subjects
with greatest
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plasma [almitrine] were most at risk. Averaged, those presenting peripheral
neuropathy in the
study year had plasma [almitrine] of 344 and 617 ng/ml at 3 and 12 months
respectively,
those that didn't had plasma [almitrine] of 249 and 387 ng/ml at 3 and 12
months
respectively. The suggested optimal long term mean (not peak) plasma
[almitrine] for treating
COPD, without inducing neuropathy, is in the 200-300 ng/ml range [266, 272].
Almitrine has
a long half-life in the human body. When daily almitrine intake exceeds daily
almitrine
elimination from the body, as it does with 2100 mg oral almitrine dimesylate
dosed per day
[263], there is fractional compounding of daily doses, which drives higher
plasma [almitrine]
over time, until an eventual point (between day 90 and day 180 with the 100 mg
administered
in [263]) when plasma [almitrine] stops increasing and stabilizes. If the
almitrine dimesylate
daily dose is sufficiently high (2100 mg and more surely 2200 mg), peripheral
neuropathy
can occur in the minority of COPD subjects that eliminate almitrine most
poorly, who have
greatest compounding of almitrine daily doses, and in which the greatest mean
and trough
plasma [almitrine] occurs (trough refers to the lowest drug concentration
inbetween two
doses). Wherein it still typically takes months, on 100-200 mg almitrine
dimesylate daily, for
this fraction of COPD subjects to accumulate enough almitrine in their body to
cause
peripheral neuropathy [272]. The majority of COPD subjects don't accumulate
such an
amount with clinically used almitrine doses (50 to 200 mg oral almitrine
dimesylate per day),
at least for the length of periods that have been studied, which is months to
years. So
peripheral neuropathy is a dose-dependent side-effect, wherein the
overwhelming majority of
COPD subjects administered, even with 200 mg oral almitrine dimesylate daily,
don't
accumulate sufficient [almitrine] in their body for it to occur. Almitrine
administered subjects
suffering paraesthesiae/peripheral neuropathy must already have a high
almitrine
concentration in their body, which is enough to exert significant anti-cancer
activity in
subjects with cancer. For these subjects, the solution is just to stop/reduce
their almitrine
daily dosage. The almitrine concentration in their body will then decrease,
and the
paraesthesiae/peripheral neuropathy with it. Afterwards, optionally, their
almitrine dosing can
be restarted, optionally at a lower (e.g. daily) dosage. This is a reactionary
dosage regime.
Alternatively a "proactive dosage regime" can be used. Wherein the
administered daily
dosage of almitrine, or a pharmaceutically-acceptable salt, solvate, hydrate
or prodrug
thereof, is decreased over the treatment course, by some function of time
since treatment start
(non-limiting e.g. 200 mg almitrine dimesylate administered per day for 1s1
month, 100 mg
per day in subsequent month(s)), and/or the course of administration has
pauses or rest
periods, wherein less or no almitrine is administered (non-limiting example: a
repeating cycle
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of [2 months almitrine administration, 1 month none]). Optionally the
almitrine
administration dosage/regime can be individually tailored to the almitrine
elimination
parameters of each individual subject. When deciding the almitrine
dosage/regime to
administer, the duration of the administration course should be a principal
consideration.
Wherein if the course is short, e.g. for days to weeks, it is probable that
constantly high daily
almitrine concentrations (e.g. 200 mg oral almitrine dimesylate per day) will
not produce
significant side-effects in most to all subjects. Indeed, 400 mg oral
almitrine dimesylate per
day is safe, in healthy subjects at least [259], at least over the short term.
For longer courses
of almitrine administration (non-limiting e.g. >3 months), to protect the
minority of subjects
susceptible to associated peripheral neuropathy, a "proactive dosage regime",
as prior
defined, can be used, and/or a lower almitrine dose. Illustratively, in human
COPD patients,
[273] 75 mg oral almitrine dimesylate administered daily for 6 months produced
no adverse
effects in any of the study subjects, wherein at study end, the average of the
trough (lowest
inbetween doses) plasma [almitrine] in the subjects was 302 ng/ml. In human
COPD patients,
[274] administered 100 mg oral almitrine dimesylate daily for 2 months, then 1
month with
no administration, and this cycle was repeated for I year, at the end of which
the mean
plasma [almitrine] was 285 ng/ml, and the authors mention that "there is a
close relationship
between the almitrine [mean or trough I would specify] plasma level and the
occurrence of
side-effects such as peripheral neuropathies", concluding that their regime is
safe. In human
COPD patients, [275] administered 50 mg almitrine dimesylate twice daily (i.e.
100 mg per
day) for 8 weeks and, before the morning dose, plasma [almitrine] was on
average across the
subjects: 93, 134, 148, 171 ng/ml on days 14, 28, 42, 56 respectively, and 104
ng/ml 2 weeks
after the study. In different human COPD patients, [275] administered 100 mg
almitrine
dimesylate twice daily (i.e. 200 mg per day) for 8 weeks and, before the
morning dose,
plasma [almitrine] was on average across the subjects: 268, 409, 442, 572
ng/ml on days 14,
28, 42, 56 respectively, and 311 ng/ml 2 weeks after the study. In human COPD
patients,
over long term, two 50 mg doses of oral almitrine dimesylate per day results
in plasma
[almitrine] that is 2 to 3 times higher than a single 100 mg oral dose [276],
so dividing the
daily dose can make a big difference. In human COPD patients, [263]
administered 100 mg
.. oral almitrine dimesylate daily for 1 year and at year end mean plasma
[almitrine] was 409.35
ng/ml, trough plasma [almitrine] was 301.8 ng/ml (doesn't comment on any side-
effects
observed or lack thereof, was a purely pharmacokinetic study). In human COPD
patients
[277], 100 mg oral almitrine dimesylate daily for I year increased the
occurrence, compared
to placebo control, of a nervous system disorder by 4%, paraesthesia by 5% and
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CA 3050553 2019-07-25
polyneuropathy by 3.6%. Anyhow, I will now discuss how almitrine associated
peripheral
neuropathy is probably more prevelant in COPD than cancer patients, thence is
less of a
concern in cancer patients.
Greater incidence of peripheral neuropathy is observed in almitrine than
placebo
administered COPD patients [265]: 9% greater chance of peripheral neuropathy
in COPD
patients administered almitrine than placebo [278]. However, a complicating
issue is that
COPD itself is associated with sub-clinical (93.8% [279], 60-87% [280], 58-95%
[281] 87%
[283], 58% [284], 4% [285] incidence in COPD patents), and even clinically
evidential (40%
[280], 7-88% [281], 87.5% [282], 17% [283], 7% [284], 20% [285] incidence in
COPD
patents), peripheral neuropathy. Plus, there is wider evidence to suggest that
hypoxia, which
is of course symptomatic of COPD, can cause peripheral neuropathy [286, 287].
So, it might
be that almitrine doesn't cause peripheral neuropathy per se, but that it
unmasks/increases
COPD driven peripheral neuropathy, perhaps in some cases elevating it from sub-
clinically
evident (e.g. in electrophysiological recordings, reduced nerve conduction
velocities etc.) to
clinically evident (patient complains of neuropathic pain etc.). Thus, I
anticipate that
almitrine used for a different disease, e.g. cancer, which isn't characterised
by poor 02
delivery to normal body tissues, will have a lower incidence of peripheral
neuropathy than its
use for COPD: much lower frequency of incidence, and much longer duration of
use
(therapeutic window), before any neuropathic onset. Thus, the risk-reward
benefit for
almitrine cancer treatment might be even greater than the present data would
at first glance
suggest.
Many COPD patients are or were smokers, often heavy. Indeed, in many patients
smoking is
the drive to their COPD. Moreover, smoking constricts blood vessels, driving
hypoxia, and
contains many harmful chemicals, and smoking and peripheral neuropathy are
correlated in
COPD patients [283] and a wider patient set (smokers reporting to the hospital
for any reason
were investigated) [288]. Few of the clinical studies, used by [257] to
appraise almitrine use
for COPD, wherein [257] recommended almitrine withdrawal, control for smoking,
past or
present, which is an oversight, and merits re-appraisal, which can't be done
in most cases
because the constituent clinical studies didn't document the smoking
frequency/history of
their particpants (at least this isn't reported in their publications).
Moreover, because the
almitrine group is larger than the placebo group in most of these
studies/pooled studies [278],
if a given percentage of trial participants smoke or have smoked (likely to be
high in COPD
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CA 3050553 2019-07-25
patients), there will be a higher percentage of trial smokers in the almitrine
than placebo
group. Nor do these almitrine clinical studies control for alcohol intake,
which also
predisposes to peripheral neuropathy (alcoholic polyneuropathy), wherein a
COPD patient set
is likely to have a disproportionate number of heavy drinkers than general
population because
smoking is a strong drive to COPD and smokers are disproportionally alcohol
drinkers e.g.
refer [289]. Nor do they control for diabetes, which correlates with COPD
(COPD patients
have higher diabetes prevelance than general population [290]), wherein
diabetes is
associated with neuropathy [286, 287].
Peripheral neuropathy incidence in almitrine dimesylate administered COPD
patients is low
(and might be even lower, as explained prior), only occurs after prolonged
administration
(mean onset = 11 months into the almitrine administration [257], therapeutic
window) and in
most cases is reversible, in the minority of cases that it occurs. Plus, it
might be even less
frequent/serious in cancer than COPD patients (as prior explained). So,
almitrine conferred
peripheral neuropathy is much less prevelant/serious than Chemotherapy Induced
Peripheral
Neuropathy (CIPN), which occurs in 71-96% [291]/52.7% [292]/68.6% [293] of
patients
who receive chemotherapy, 68.1% have it from the 1st month of chemotherapy
treatment
[294], wherein CIPN is irreversible in many cases [292], greatly diminishing
the quality of
life of cancer survivors (cancer patients rank CIPN near the most intense pain
imaginable, 8
or 9 on a 10-point scale, "like walking on shards of glass or hot coals"
[295]). Carboplatin
produces peripheral neuropathy in 49% of cancer patients [292], whereas
almitrine
dimesylate produces a much lower incidence of peripheral neuropathy
(reversible, only after
prolonged use), wherein almitrine dimesylate exerts greater anti-cancer
activity in
standardised testing (at 10 NI) than carboplatin (compare Figures 1 and 7A):
almitrine =
less side-effects + more potent anti-cancer activity = better anti-cancer
drug. Furthermore,
chemotherapies deliver many further, horrific side-effects, which almitrine
doesn't. Indeed, in
some cases chemotherapy, rather than cancer, kills cancer patients [296]. That
is how toxic
chemotherapies in present use are. Almitrine dimesylate, with its anti-cancer
activity and low
side-effect profile vs. chemotherapy, will help many cancer patients. For
example, firstly, the
many that withdraw from chemotherapy because of its horrific side-effects and
whose present
prognosis is stark. With the widespread problem of cancer resistance to
present
chemotherapies, which kills many, any new cancer drug is a blessing,
especially one so
structurally distinct, and targeting a target so different, from other cancer
drugs, as is the case
with almitrine.
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CA 3050553 2019-07-25
For the minority of almitrine administered patients that do present peripheral
neuropathy, and
for those that don't (i.e. proactive treatment), an invention embodiment is to
administer a
treatment(s) sanctioned for treating peripheral neuropathy. For (non-limiting)
example, a
.. treatment(s) that is, or has been, used for CIPN and/or diabetic neuropathy
e.g. (non-limiting)
one or more of duloxetine (a preferred embodiment, [297]), a tricyclic
antidepressant(s),
pregabalin, venlafaxine, carbamazepine, oxcarbazepine, amifostine, Org 2766,
zonisamide,
gabapentin, lamotrigine, prednisone, baclofen, am itriptyline, ketamine,
intravenous calcium
and/or magnesium, glutathione, amifostine, glutamine, acetyl-L-carnitine,
alpha-lipoic acid,
acetylcysteine, diethyldithiocarbamate, vitamin E, vitamin B6, valproate,
interleukin-6,
capsaicin cream, tapentadol, lidocaine patches, topical menthol, a topical
analgesic(s), an
alpha-2-delta antagonist(s), an antiepileptic/anticonvulsant drug(s) (AEDs), a
serotonin-
norepinephrine reuptake inhibitor(s) (SNRIs), a tricyclic antidepressant(s)
(TCAs), an
opoid(s), morphine sulfate, oxycodone, botulinum toxin, acupuncture, scrambler
therapy,
cannabis, THC, transcutaneous electrical nerve stimulation (TENS),
interferential current
(IFC), medical device(s).
Almitrine's anti-cancer activity was unknown prior to my work, despite
almitrine being
around since the early 1970s, approaching 50 years ago. Its anti-cancer
activity is unexpected
to a person of the art. Especially because another respiratory stimulant,
doxapram, has been
publically shown by others to have no anti-cancer activity in the same one-
dose (10 M)
NCI-60 test in which, disclosed herein, almitrine dimesylate (10 M) exerts
anti-cancer
activity. Doxapram in NCI-60 (10 pM) testing: mean % cancer growth inhibition
= -3.7%
(median = -2.3%) i.e. negative numbers show cancer growth promotion (!) rather
inhibition,
as compared to no drug control, NSC: 760347 in [16]. Thence, the discovery of
almitrine
conferred anticancer activity, disclosed herein, is unforeseen by a person of
the art, novel and
componentry to the invention of this disclosure. The risk-reward axis for
almitrine is
sufficient, indeed it is excellent, for an anti-cancer drug. Especially when
used acutely,
because most of almitrine's side-effects only occur with chronic use. Acute
almitrine use for
.. cancer treatment has a different risk-reward axis than chronic almitrine
use for COPD
treatment (for which it is ineffective [257]), especially because almitrine's
side-effects are
mostly associated with chronic use, and because cancer can be an immediately
life-
threatening disease for too many patients, with too few life-saving options,
wherein present
treatments typically have absolutely horrendous side-effects ("after a few
courses, Tim only
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CA 3050553 2019-07-25
needed to see the drip coming towards him down the corridor to trigger
aggressive and
relentless vomiting", Cancer Research UK article on "Our milestones:
Cisplatin" [298], one
of, if not the most, widely used cancer drug today). Indeed, the merit of anti-
cancer treatment
merits the risk of higher almitrine dosages than 200 mg per day.
Intravenous delivery of 459 155 mg almitrine dimesylate, infused within 24
hours, caused
reversible lactic acidosis and hepatic dysfunction in 30% of 25 patients
[299]. The other 70%
of patients had no ill effects, and unaltered plasma [lactate]. The side-
affected minority
correlated with an impaired liver function parameter, increased plasma
[bilirubin], prior to
almitrine administration. Thus, this side-affected cohort is largely
predictable. Most side-
affected were women, but not all women were affected (N.B. women can have a
smaller liver
relative to body size e.g. refer [300]). The liver converts lactate to glucose
by the Cori cycle
[1] and an impaired/overwhelmed liver cannot process the elevated plasma
lactate that
almitrine administration can cause [252], which renders lactate acidosis. An
embodiment of
this invention is to select a cancer patient's almitrine dosage dependent upon
their liver
function. That is, in a further (non-limiting) embodiment, assessed by
measuring plasma
[bilirubin]. For non-limiting example: if (plasma [bilirubin] > 17 M) {the
patient should not
be administered high almitrine dosage(s)}. Patients with better liver function
are at less risk
of almitrine driven lactic acidosis [299] and can endure higher almitrine
dosages. Another
embodiment is to record plasma [lactate], and/or a liver function assay
chemical(s) (non-
limiting e.g. bilirubin), whilst a cancer patient is administered with
almitrine, or a course of
almitrine administrations, and to lower the administered almitrine
dosage/frequency if these
plasma concentrations become abnormal. An invention embodiment is to use
almitrine, and a
drug(s)/treatment(s) treating/mitigating lactic acidosis (e.g. sodium
dichloroacetate, DCA,
which also has reported anti-cancer activity [47]), in anti-cancer therapy.
Another
embodiment is almitrine for anti-cancer therapy, given with a dosage
adjustment dependent
upon initial body weight, before treatment, and optionally reducing the dosage
if significant
weight loss occurs. Almitrine in co-therapy with a high(er) calorie diet, as
an anti-cancer
treatment, is another embodiment. Almitrine in co-therapy with a drug to treat
or mitigate
peripheral neuropathy (e.g. {non-limiting} gabapentin, duloxetine, pregabalin
etc.), as an
anti-cancer treatment, is an embodiment of this invention. An embodiment is to
use almitrine
for anti-cancer therapy and to monitor the almitrine recipient, or for the
almitrine recipient to
self-monitor, for weight loss and/or signs of neuropathy, and/or odd
neurological sensations,
especially in the body periphery e.g. the limbs. An embodiment is to use
almitrine for anti-
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cancer therapy under medical supervision. Wherein, in animal or human,
almitrine dosage,
frequency, route and duration of administration is directed/recommended,
and/or almitrine is
administered, by a medically qualified professional(s) e.g. a doctor or vet or
nurse or
pharmacist. In an embodiment, an oncologist or other cancer specialist or a
medically
qualified professional that has undergone additional training and/or
qualification and/or
residency in oncology beyond a degree in human and/or veterinary medicine. And
optionally
wherein one or more of the dosage, frequency, route and duration of almitrine
administration
is modulated in the light of cancer progression/regression/stasis during the
course of almitrine
administration.
Mechanistic studies in animals [301-302] have identified that it might not be
almitrine itself
that causes almitrine associated neuropathy but instead
difluorobenzhydrylpiperadine
(DFBP), which is the major almitrine metabolite formed in humans. DFBP also
causes
weight loss in [301] and so DFBP could also be the basis to almitrine
associated weight loss
(reported in [257]), or this could just be a function of altered feeding
behaviour as a function
of the DFBP generated neuropathy. To render DFBP from almitrine, the bond
between
almitrine's nitrogen, at atom number 11, and carbon, at atom number 9, must be
broken. An
embodiment of this invention is almitrine isotopically enriched (greater than
natural
abundance, e.g. {non-limiting} >70%) for 15N at Atom Number 11, and/or
isotopically
enriched for '3C at Atom Number 9 (can be done, to illustrate and not limit,
by substituting
cyanuric chloride-'3C3 {CAS: 286013-07-8; available from Sigma-Aldrich} for
cyanuric
chloride in almitrine synthesis, to give almitrine enriched at three carbon
positions with '3C,
including at Atom Number 9), which will make this bond stronger by the kinetic
isotope
effect (KIE), which will reduce the rate of DFBP formation, and reduce
neuropathy (Atom
Numbers as labelled by [25]). Kinetic isotope effect (KIE) is the change in
the rate of a
chemical reaction when one (or more) of the atoms in the reactants is replaced
with its
isotope. Heavier isotopes form stronger bonds that require higher energy to
break them,
which ultimately slows down the chemical reaction rate. Other atom(s) of
almitrine enriched
(greater than natural abundance, e.g. {non-limiting} >70%) with their heavier,
stable
respective isotope(s) (e.g. {non-limiting} 2H replacements of 'H) is also
componentry to the
present invention. As is one or more hydrogen atom(s) upon almitrine, or an
aforementioned
almitrine isotopologue, replaced by fluorine (or other halogen), especially
near the N11-C9
bond that breaks to release DFBP, preferably upon the piperazine ring. The use
of one or
more of the new compositions of matter of this disclosure, to treat a
condition for which
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almitrine has been used in humans, for (non-limiting) example, chronic
obstructive
pulmonary disease (COPD), is componentry to this invention. As is their use as
an anti-
cancer treatment.
Following reactions are illustrative, not restrictive: almitrine could be
deuterated, upon its
piperazine ring and/or other loci, by reactions described in [N], which
deuterate sp3 carbons.
And/or by reactions described in [RI, R2, Q1, Q2] which deuterate widely, upon
aromatic
and alkyl molecular components. And/or by reactions described in [01, 02],
which deuterate
a- and 0-carbons to phenyl groups. And/or by reactions described in [D], which
deuterate a-
and 0-carbons to tertiary amines. And/or by reactions described in [F, El, E2,
Exl], which
deuterate a-carbons to tertiary amines. And/or by reactions described in [A,
B, El, E2, F],
which deuterate a-carbons to secondary amines. Whichever option(s) is chosen,
solvents,
temperatures, pressures, and other reaction conditions can be selected by one
of ordinary skill
in the art. Deuteration can be modulated by modulating reaction time: greater
deuterium
incorporation by longer reaction time. One can do multiple cycles of one or
more of these
reactions until the desired level of deuterium incorporation occurs, monitored
by 1H and/or
2H NMR and/or mass spectrometry.
Encompassed by this invention are methods of administering an effective amount
of almitrine
(and/or one or more of its metabolites or derivatives), or a pharmaceutically-
acceptable salt,
solvate, hydrate or prodrug thereof, or a pharmaceutical composition(s)
comprising almitrine
(and/or one or more of its metabolites or derivatives), optionally in co-
therapy with another
anti-cancer treatment(s) including radiotherapy, to
treat/ameliorate/prevent/combat cancer in
a subject. Encompassed by this invention are methods of administering an
effective amount
of GAL021 [268-269] and/or any compound(s) of [P8-12], or a pharmaceutically-
acceptable
salt, solvate, hydrate or prodrug thereof, or a pharmaceutical composition(s)
comprising
GAL021 [268-269] and/or any compound(s) of [P8-12], optionally in co-therapy
with another
anti-cancer treatment(s) including radiotherapy, to
treat/ameliorate/prevent/combat cancer in
a subject. Encompassed by this invention are methods of administering an
effective amount
of doxapram, or a pharmaceutically-acceptable salt, solvate, hydrate or
prodrug thereof, or a
pharmaceutical composition(s) comprising doxapram, optionally in co-therapy
with another
anti-cancer treatment(s) including radiotherapy, to
treat/ameliorate/prevent/combat cancer in
a subject. Encompassed by this invention are methods of administering an
effective amount
of a (e.g. chemoreceptor) respiratory stimulant(s), or a pharmaceutically-
acceptable salt,
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CA 3050553 2019-07-25
solvate, hydrate or prodrug thereof, or a pharmaceutical composition(s)
comprising a
respiratory stimulant(s), optionally in co-therapy with another anti-cancer
treatment(s)
including radiotherapy, to treat/ameliorate/prevent/combat cancer in a
subject. Encompassed
by this invention are methods of administering an effective amount of a
compound(s) that
increases p02 in the subject's blood, or a pharmaceutically-acceptable salt,
solvate, hydrate
or prodrug thereof, or a pharmaceutical composition(s) comprising a
compound(s) that
increases p02 in the subject's blood, optionally in co-therapy with another
anti-cancer
treatment(s) including radiotherapy, to treat/ameliorate/prevent/combat cancer
in the subject.
A method of treating, ameliorating, preventing or combating cancer in a
subject wherein the
method comprises the subject taking, or being administered, a therapeutically
effective
amount of almitrine and/or other compound(s) of Formula VI (and/or a
pharmaceutical
composition(s) containing a therapeutically effective amount of almitrine
and/or other
compound(s) of Formula VI herein). Almitrine and/or other compound(s) of
Formula VI,
.. (and/or a pharmaceutical composition(s) containing almitrine and/or other
compound(s) of
Formula VI) for use in the treatment/amelioration/prevention/combat of cancer
in a subject.
The use of almitrine, and/or other compound(s) of Formula VI, in the
manufacture of a
medicament for the therapeutic and/or prophylactic treatment of cancer,
optionally in a ready-
to-use drug form, optionally in a package together with instructions for its
anti-cancer use.
Almitrine, and/or other compound(s) of Formula VI, for use in a method for the
treatment/amelioration/prevention/combat of cancer and/or ischemia and/or
stroke (reduces
ATP hydrolysis and maintains intracellular [ATP] when 02 and glucose is in
short supply
because of a vascular occlusion or similar) in a subject.
An invention embodiment is a pharmaceutical composition comprising a
therapeutically
effective amount of almitrine and a fatty acid(s), wherein the scope of what
is a fatty acid is
well known to those of the art. For non-limiting example, wherein almitrine
and a fatty acid
are in a 1:2 ratio. Other stoichiometries/ratios are also componentry to the
present invention.
For example, a 1:1 almitrine to fatty acid ratio. Some fatty acid(s) can exert
anti-cancer
activity (illustrative literature: [303, 304]) and in preferred embodiments
almitrine is
combined in a composition(s) with a fatty acid(s) that exerts anti-cancer
activity, and in more
preferred embodiments the anti-cancer activity of almitrine and fatty acid(s)
synergise. The
greater the anti-cancer activity of the fatty acid(s) the more preferred the
embodiment of its
formulation with almitrine. In a further embodiment, an almitrine and fatty
acid(s) containing
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CA 3050553 2019-07-25
composition(s) is used in a method of treatment of the human or animal body by
therapy, for
(non-limiting) example to treat/ameliorate/prevent/combat cancer in a subject.
In another
embodiment, an almitrine and fatty acid(s) containing composition is used for
the
manufacture of a medicament for the treatment/amelioration/prevention/combat
of cancer. An
invention embodiment is almitrine in a 1:2 stoichiometry with 9Z,1 1E
conjugated linoleic
acid (Rumenic Acid), which is an example of a fatty acid with anti-cancer
activity [lips2]. In
other illustrative example embodiments almitrine is in a 1:2 stoichiometry
with
Eicosapentaenoic Acid, or Docosahexaenoic Acid, or Erucic acid. Example
embodiment:
FJF
H30,,
01-i3
N N
o
H20
N N N
OH
HO
0
In some embodiments, almitrine or a pharmaceutically-acceptable salt, solvate,
hydrate or
prodrug thereof is used as an adjuvant or neoadjuvant to another cancer
treatment(s) e.g. used
as an adjuvant to chemo and/or radiotherapy e.g. used as a chemosensitizer
and/or
radiosensitizer/radioenhancer. Many conventional [chemo/radio] therapies act
against cancer,
wholly or in part, by increasing [ROS]. Indeed, radiotherapy [305] and
chemotherapy [306,
307, 308] increase [ROS] in cancer cells. A mechanism that cancers use to
mitigate and
counteract this [ROS] increase is greater FiFo ATP hydrolysis, consuming ATP,
which
releases glycolysis from ATP feedback inhibition, permitting higher glycolytic
and pentose
phosphate pathway rate, more NADPH produced and thence greater ROS mitigation.
Almitrine slows Fi Fo ATP hydrolysis, reducing ROS mitigation and this assists
the anti-
cancer activity of chemo/radio therapy. Almitrine also increases blood and
tissue p02,
increasing ROS production. So, almitrine combats tumor hypoxia, wherein this
hypoxia can
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CA 3050553 2019-07-25
be a drive to radio- [305] and chemo- [sens5, sens6] resistance of cancer. In
some
embodiments the anti-cancer activity of almitrine or a pharmaceutically
acceptable salt,
solvate, hydrate, prodrug thereof, synergises with (potentiates) the anti-
cancer activity of an
FDA and/or EMA approved anti-cancer treatment(s) e.g. one or more of
chemotherapy,
radiotherapy, immunotherapy, surgery, immune-oncology, radioimmunotherapy,
biological
therapy, hormone therapy etc. In other words, in some embodiments, the
combined anti-
cancer effect of almitrine and another cancer treatment(s) is greater than the
sum of each
alone. In some embodiments, almitrine administration enables the same or
greater anti-cancer
activity to be exerted by another anti-cancer treatment(s) but with lower
radiative (e.g. x-ray,
y ray, electromagnetic radiation, radioactivity etc.) and/or drug(s) exposure,
e.g. lower radio-
and/or chemo- therapeutic(s) dose, most preferably enabling a lower side-
effect profile.
Encompassed herein is a method of reducing, treating and/or preventing adverse
or undesired
effects associated with conventional therapy including, but not limited to,
chemotherapy,
radiotherapy, immunotherapy, wherein almitrine or a pharmaceutically
acceptable salt,
solvate, hydrate, prodrug thereof, is administered to a subject prior to,
during, or after the
occurrence of the adverse effect associated with conventional therapy,
optionally wherein the
dosage/frequency/use of the conventional therapy is decreased. In some
embodiments,
almitrine is used in co-therapy with cisplatin and/or carboplatin and/or some
other platinum
based therapeutic(s) for anti-cancer treatment in a subject, and in further
embodiments their
anti-cancer activities synergize. In some embodiments, almitrine or a
pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof is used in co-therapy
with radiotherapy for
anti-cancer treatment in a subject, and in further embodiments their anti-
cancer activities
synergize. In some embodiments, almitrine or a pharmaceutically-acceptable
salt, solvate,
hydrate or prodrug thereof is administered alongside radiotherapy to treat
radioresistant
cancer(s) and/or alongside chemotherapy to treat chemoresistant cancer(s).
In some embodiments, almitrine or a pharmaceutically-acceptable salt, solvate,
hydrate or
prodrug thereof is administered/prescribed for anti-cancer therapy in a
subject(s) at King's
College Hospital, London, and/or another National Health Service (NHS)
hospital/location
and/or by an NHS employee(s) and/or in a country where almitrine or a
pharmaceutically-
acceptable salt, solvate, hydrate or prodrug thereof is, or is not, available
for
prescription/administration by a medical practitioner(s). The i.v. dose of
almitrine used in
clinical practice at King's College Hospital (London, UK) is 8 tig/kg/min for
COPD and 4-16
tig/kg/min for Acute Respiratory Distress Syndrome (ARDS) [271]. Intravenous,
as opposed
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CA 3050553 2019-07-25
to oral, administration of almitrine or a pharmaceutically-acceptable salt,
solvate, hydrate or
prodrug thereof can be advantageous for anti-cancer therapy in a subject,
especially in
clinical trialing, because subject variability in oral bioavailability of
almitrine is rendered
irrelevant. An aspect to this is that, unlike oral administration, i.v.
dosing, if sufficiently
large, permits one to know precisely when peak plasma [almitrine] occurs in
each subject. It
being at the final time point of i.v. administration. This is useful when
almitrine or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof is
administered with
radio- and/or chemo- therapy and maximal therapeutic synergy is desired. For
example, if
radiotherapy can't be administered during, it should be administered as soon
as possible after,
the i.v. administration of almitrine or a pharmaceutically-acceptable salt,
solvate, hydrate or
prodrug thereof. The shorter the delay between the end of i.v. almitrine
administration and
the start of radiotherapy, the more preferred the embodiment. Most preferred
is the delay is
zero and they are concurrent. During concurrent administration, radiotherapy
preferably starts
after the start of i.v. almitrine administration, to permit a build up of
[almitrine] in the tissues
.. before radiotherapy commences.
In humans, [266] infused ¨7.47 g/kg/min for 2 hours, delivering 60 mg
almitrine
dimesylate, wherein the mean almitrine plasma level in the subjects was 327
ng/ml at the end
of the infusion period (lowest observed = 242 ng/ml), which then fell to to
157, 154, 105, 67,
55 ng/ml, at 15, 30, 60, 120, 600 minutes afterwards respectively. In humans,
[312] infused 8
g/kg/min for 20 minutes during which the mean plasma concentration of
almitrine was 325
ng/ml. In humans, [313] infused 8.3 g/kg/min for 30 minutes, wherein "no
adverse side-
effect was observed during or after the administration of almitrine". In
humans, [314] infused
16 g/kg/min for 1 hour, during which plasma almitrine concentration rose to
be greater than
600 ng/ml in all subjects, much greater in some subjects (-1,600 ng/ml highest
observed),
and by 2 hours after the infusion stopped, this had fallen to below 400 ng/ml
in all subjects
but one, and by 12 hours was below 200 ng/ml in all. In humans, [315] infused
16.7
g/kg/min for 1 hour "without deleterious effects". In humans, [316] infused 16
g/kg/min
= and 20 minutes into this infusion the mean plasma concentration of
almitrine was 659 ng/ml,
wherein "plasma lactate concentrations remained within the normal range in all
patients".
There are many more human studies in the literature wherein ¨16 or 16
g/kg/min is infused,
and many more wherein less is infused. Someone of the art will know how to
find all these. A
fraction, but by no means all, are listed and usefully compared in a table in
[270]. In humans,
[3 1 7] infuses a higher rate than ¨16 g/kg/min, = 25 g/kg/min for 1 hour.
[270], a French
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CA 3050553 2019-07-25
regulatory review document for injectable almitrine, states "maximum
recommended flow-
rate: 15 mg/minute", which corresponds to a flow rate of 242 g/kg/min for a
62 kg human.
In humans, [318] gave a rapid intravenous (bolus) injection of 0.5 mg/kg,
followed by
infusing 2 mg/kg (corresponds to 16.7 g/kg/min for a 62 kg human) for 2
hours, giving 2.5
mg/kg (corresponds to 155 mg for a 62 kg human) in just ¨2 hours. 200 mg
(=3.23 mg/kg for
a 62 kg human) almitrine dimesylate orally per day has clinical precedent
[257], as does an
i.v. infusion rate of 16 g/kg/min [271]. Combining these, one arrives at a
daily i.v. infusion
of 16 g/kg/min for 202 minutes (3 hours and 22 minutes) to deliver 200 mg to
a 62 kg
human. Because, on average across subjects, only 85% [260] of the oral dose is
bioavailable,
.. this oral dose can actually be approximated (in a 62 kg human) by a daily
i.v. infusion of 16
g/kg/min for 171 minutes (2 hours and 51 minutes). To illustrate, and not
restrict, to deliver
this same dose (to a 62 kg human) of almitrine dimesylate by continuous i.v.,
with an
infusion rate of 8, 32, 64 g/kg/min would take 5.7, 1.43 and 0.71 hours
respectively.
Optionally, infusion time can be shortened by injecting a proportion of the
daily dose as i.v.
bolus, optionally preceding the steady infusion. So, to illustrate, if 15 mg
was administered
by a preceding i.v. bolus (permissible by "maximum recommended flow-rate"
instruction of
[2701) then the duration required (in a 62 kg human) to replicate a 200 mg
oral dose
(factoring in bioavailability issue) would be 159 minutes of 16 g/kg/min
continuous i.v.
infusion. In some invention embodiments, almitrine or a pharmaceutically-
acceptable salt,
solvate, hydrate or prodrug thereof is administered orally and by i.v. in
combination to
treat/ameliorate/prevent/combat cancer in a subject. So, to illustrate (not
restrict), per day,
100 mg is administered orally and 100 mg is administered intravenously by 16
g/kg/min for
(in a 62 kg human) 101 minutes, 86 minutes if 15 mg of the i.v. dose is given
by bolus.
Optionally, the oral and i.v. administrations are timed so that as the i.v.
administration
finishes, and as plasma [almitrine] from the i.v. dose declines, almitrine
from the oral dose
enters the bloodstream to buoy/increase plasma [almitrine]. Figure 3.3. in
[266] shows (data
normalised to a 1 mg/kg dose) that the gradient of increase in plasma
[almitrine] in the first 3
hours after an oral almitrine dose is similar to the gradient of decrease in
plasma [almitrine]
in the first 3 hours after an i.v. dose of almitrine terminates. So,
administering oral almitrine
.. as i.v. almitrine administration finishes can act to buoy/increase plasma
[almitrine] for a
longer period of time. This is very useful if the subject is to undergo radio-
and/or chemo-
therapy afterwards, wherein a high plasma [almitrine] is desired to gain the
maximal
additive/synergistic anti-cancer effect. All aforementioned dosages,
routes/patterns of
administration and infusion rates of almitrine or a pharmaceutically-
acceptable salt, solvate,
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CA 3050553 2019-07-25
hydrate or prodrug thereof, in use for anti-cancer therapy/treatment in a
subject, are
componentry to the present invention. As are others, which are also
contemplated by the
scope of this invention. For example, the administered daily dosage could be
greater than 200
mg. Indeed, relevantly, 400 mg oral almitrine dimesylate per day has been
shown safe in
healthy subjects, at least for the duration of that study [259]. Or the
administered dosage
could be less.
Intravenous infusion of 5.5 ( 1.7) g/kg/min almitrine dimesylate, in co-
therapy with
inhalation of nitric oxide (5 p.p.m.), increased arterial p02 by >30% across
all subjects [311].
There are many similar papers in the literature, describing co-administration
of almitrine and
NO, which can be found readily by someone of the art. In some invention
embodiments,
almitrine or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug
thereof is
administered, optionally intravenously, in co-therapy with nitric oxide (NO),
optionally
breathed, to treat/ameliorate/prevent/combat cancer in a subject, optionally
in co-therapy with
.. radio- and/or chemo- therapy, wherein the almitine and NO combination
increases the
subject's blood and tissue p02, which makes radio- and/or chemo- therapy more
effective,
adding to/synergising with the inherent anti-cancer activity of almitrine. At
any point that
almitrine administration is referred to in the disclosure, in further
embodiments of this
invention, NO is administered also.
Componentry to this invention is administering an i.v. bolus dose of almitrine
or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof for anti-
cancer therapy
in a subject, optionally wherein the subject has lung cancer, and optionally
wherein the
almitrine concentration in the body is subsequently increased/prolonged by a
subsequent
period(s) of continuous i.v. infusion (optionally where the infusion rate
equals/approximates
the almitrine elmination rate) and/or one or more oral administrations of
almitrine or a
pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof (non-
limiting e.g. tablet
and/or solution). Optionally, wherein a foundational concentration of
almitrine in subject
plasma has been built up, prior to the i.v. dose of almitrine or a
pharmaceutically-acceptable
salt, solvate, hydrate or prodrug thereof, by one or more oral and/or i.v.
doses of almitrine or
a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
optionally
administered on the same day and/or at some regular/irregular frequency, e.g.
daily, in the
day(s)/week(s)/month(s) before. Faster almitrine build up in the body can be
achieved by
dividing the daily dose up into multiple smaller doses, e.g. (non-limiting)
200 mg per day is
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administered by 100 mg administered twice per day, which means the build up
period can be
shorter.
At every point that radiotherapy is referred to in this disclosure, it
encompasses all that a
person of the art would expect it to. Including, without limitation, external
(including,
without limitation, one or more of external beam radiation therapy {e.g. using
photons/electrons/hadrons/protons/neutrons/ions/nuclei etc.}, stereotactic
body radiation
therapy [SBRT], radiosurgery, 3-dimensional conformal radiation therapy, image-
guided
radiation therapy, intensity-modulated radiation therapy, tomotherapy,
volumetric modulated
arc therapy, particle therapy, proton therapy, neutron capture therapy, auger
therapy) and/or
internal (including, without limitation, one or more of brachytherapy,
unsealed source
radiotherapy, intraoperative radiotherapy, deep inspiration breath-hold,
selective internal
radiation therapy) radiotherapy. Moreover, when radiotherapy is referred to in
this disclosure
it encompasses radiotherapy with or without the co-administration of excess
oxygen, wherein
the subject breathes gas with a greater 02 fraction than normal air at that
altitude, optionally
pure 02, optionally the subject is administered hyperbaric 02 therapy. An
embodiment of this
invention is to administer almitrine or a pharmaceutically acceptable salt,
solvate, hydrate,
prodrug thereof to a subject with cancer, optionally lung cancer, either
orally and/or by i.v.
(bolus and/or continuous) and/or by some other administration route, prior (on
the same day
and/or on prior day[s]) and/or during and/or after radiotherapy and/or
chemotherapy. In some
embodiments, almitrine or a pharmaceutically acceptable salt, solvate,
hydrate, prodrug
thereof is i.v. administered (bolus and/or continuous) before, during and
after the subject
undergoes radiotherapy and/or chemotherapy (optionally administered by i.v.
also, bolus
and/or continuous, in a separate or same infusion line to the subject),
alternatively only
before, or only during or only after, alternatively only before and after,
alternatively only
before and during, alternatively only during and after. In some embodiments,
the
administered almitrine enables a lower radiative/ionizing/chemotherapeutic(s)
dose to be
used to convey therapy. Optionally, it permits the same radiative dose to be
used but over a
longer timeframe, so the radiative intensity (per unit time) is less.
Alternatively, the same
radio- and/or chemo- therapy dose is used and greater therapeutic effect
ensues. In a further
embodiment, the subject has cancer and in more particular embodiments, the
subject has lung
cancer. In some invention embodiments a high g/kg/min i.v. infusion rate,
e.g. >8
g/kg/min or ?16 g/kg/min or _>_64 mg/kg/min, of almitrine or a
pharmaceutically acceptable
salt, solvate, hydrate, prodrug thereof is delivered to a subject, optionally
that has cancer,
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CA 3050553 2019-07-25
optionally lung cancer, prior and/or during and/or after radiotherapy and/or
chemotherapy. A
high infusion rate (by bolus only, or bolus+continuous, or continuous only)
optimizes for all,
or a significant proportion of, the daily almitrine dose, which in an
embodiment is 200 mg but
in other embodiments is higher or lower, being inside the subject at or around
the time that
radiotherapy and/or chemotherapy is administered, which gives the best
opportunity for
therapeutic synergy to occur. In some embodiments, i.v. (bolus and/or
continuous) almitrine
or a pharmaceutically acceptable salt, solvate, hydrate, prodrug thereof is
administered
preceding radio- and/or chemo- therapy and when this i.v. administration is
stopped an oral
dose of almitrine or a pharmaceutically acceptable salt, solvate, hydrate,
prodrug thereof is
administered so that the amount of almitrine entering the bloodstream from the
oral dose
partially/completely/exceeds the amount of i.v. dose almitrine eliminated from
the body,
which buoys the plasma almitrine concentration, which gives greater
opportunity for anti-
cancer therapeutic synergy between almitrine and radio- and/or chemo- therapy.
In some
embodiments, on days that the subject is administered radio- and/or chemo-
therapy, all or
some of the daily dose of almitrine or a pharmaceutically acceptable salt,
solvate, hydrate,
prodrug thereof is administered by i.v. (bolus and/or continuous), preferably
near in time to
the radio- and/or chemo- therapy, the nearer in time the better, most
preferably coincident in
time (for continuous i.v. infusion) or just before (for i.v. bolus only) or
starting just before
(for i.v. bolus followed my continuous i.v. infusion and/or oral
administration), and,
optionally, on days that the subject is not administered radio- and/or chemo-
therapy, the
daily dose of almitrine or a pharmaceutically acceptable salt, solvate,
hydrate, prodrug
thereof is absent or administered/taken orally, which the subject can do
easily outside of a
medical facility e.g. at home. So, for non-limiting example, during a daily
course of
radiotherapy and/or chemotherapy the subject receives almitrine or a
pharmaceutically
acceptable salt, solvate, hydrate, prodrug thereof by i.v. (bolus and/or
continuous) and when
this course of radiotherapy and/or chemotherapy pauses or finishes, and/or
before it starts, the
subject receives almitrine or a pharmaceutically acceptable salt, solvate,
hydrate, prodrug
thereof, optionally daily, orally, for a duration decided by a medical
practitioner(s) and/or the
subject.
For i.v. administering almitrine or a pharmaceutically acceptable salt,
solvate, hydrate,
prodrug thereof, and/or one or more chemotherapeutics (e.g. cisplatin,
carboplatin etc.),
during radiotherapy an uncharacteristically long i.v tube is preferable to
keep the bulk of the
i.v. equipment further away from the radiotherapy beam, most preferably
wherein it is located
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behind some shielding. Optionally its radioactivity, or lack thereof, is
regularly checked (e.g.
using a Geiger counter and/or using some radioactivity sensor that stays on
the equipment
always {e.g. similar to that worn by workers that work in radioactive risk
settings e.g. a
dosimeter)), in an embodiment, before each use, and if too high, it is
replaced with new
equipment. In some embodiments, radioactive/electromagnetic/ionizing shielding
is
incorporated into the equipment itself. Preferably, the i.v. administration
site(s) upon the
subject shouldn't be directly under the radiotherapy beam and should be
appropriately
shielded as much as possible. In some embodiments, multiple i.v. lines are
incident upon the
subject undergoing radiotherapy, optionally wherein different
chemotherapeutics are
adminstered by the different lines, optionally wherein one or more i.v. lines
administer
almitrine or a pharmaceutically acceptable salt, solvate, hydrate, prodrug
thereof.
In some embodiments, almitrine or a pharmaceutically acceptable salt, solvate,
hydrate,
prodrug thereof is i.v. administered (bolus and/or continuous), optionally in
the same i.v,
infusion, with one or more chemotherapies to a subject with cancer, optionally
a lung cancer
patient. In some embodiments, the timing of administering almitrine or a
pharmaceutically
acceptable salt, solvate, hydrate, prodrug thereof (e.g. oral or i.v. [bolus
and/or continuous]),
and the timing of administering one or more chemotherapies (e.g. oral or i.v.
[bolus and/or
continuous]), is coordinated so that peak plasma almitrine concentration
occurs at the same
time as the peak plasma concentration of chemotherapeutic(s), whereupon
optionally, at this
peak time, radiotherapy is administered.
In some embodiments, almitrine or a pharmaceutically acceptable salt, solvate,
hydrate,
prodrug thereof is orally administered before and/or during and/or after the
subject, who in a
further embodiment has cancer, optionally lung cancer, undergoes radiotherapy
and/or
chemotherapy; most preferably this oral almitrine is administrated before
radiotherapy and/or
chemotherapy and in further embodiments the timing of this oral almitrine
administration vs.
timing of radiotherapy and/or chemotherapy is co-ordinated so that the peak
plasma
concentration of almitrine occurs during or near in time to the radiotherapy
and/or
chemotherapy (N.B. in human, peak plasma concentration of almitrine occurs 3.5
0.7 hours
after consuming almitrine dimesylate, absorption is improved by eating food
[266]). In
further embodiments, an oral course (e.g. daily, or other frequency) of
almitrine
administration, or a pharmaceutically acceptable salt, solvate, hydrate,
prodrug thereof, is
started days/weeks/months before a course of radiotherapy and/or chemotherapy
starts, to
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build up a foundational level of almitrine inside the subject, and in further
embodiments oral
almitrine or a pharmaceutically acceptable salt, solvate, hydrate, prodrug
thereof continues to
be administered during the course of of radiotherapy and/or chemotherapy and,
in further
embodiments, afterwards also. Before radio- and/or chemo- therapy, the number
of days and
the timing/frequency/dose of almitrine administered, or a pharmaceutically
acceptable salt,
solvate, hydrate, prodrug thereof, is optimized so that the plasma
concentration of almitrine in
the subject is suitably high (in some embodiments, to illustrate and not
restrict, >300 ng/ml
and/or >200 ng/ml) before radio- and/or chemo- therapy starts, wherein in a
further
embodiment the plasma concentration of almitrine in the subject is recorded to
make sure. If
not, the same or increased dose of almitrine or a pharmaceutically acceptable
salt, solvate,
hydrate, prodrug thereof is administered/taken to/by the subject, optionally
more frequently,
for a further time period before radio- and/or chemo- therapy commences.
In some invention embodiments, a therapeutically effective amount of almitrine
or a
pharmaceutically acceptable salt, solvate, hydrate, prodrug thereof, and/or an
almitrine
containing pharmaceutical composition, is administered to
treat/ameliorate/prevent/combat
cancer in a subject, optionally lung cancer, optionally Non-Small Cell Lung
Cancer
(NSCLC), optionally lung adenocarcinoma, optionally a PET positive cancer,
optionally in
co-therapy with radiotherapy, optionally wherein almitrine makes the cancer
more
radiosensitive/less radioresistant and/or the anti-cancer activities of
almitrine and
radiotherapy add/synergize, optionally in co-therapy with one or more
chemotherapies,
optionally wherein almitrine makes the cancer more chemosensitive/less
chemoresistant
and/or the anti-cancer activities of almitrine and chemotherap[y/ies]
add/synergize, optionally
in co-therapy with (without limitation) one or more of cisplatin, carboplatin,
etoposide,
gemcitabine, vincristine, vinblastine, paclitaxel, albumin-bound paclitaxel,
vinorelbine,
topotecan, irinotecan, temozolomide, docetaxel, pemetrexed, erlotinib,
brigatinib, gefitinib,
lorlatinib, afatinib, alectinib, denosumab, ipilimumab, pembrolizumab
(Keytruda), nivolumab
(Opdivo), atezolizumab (Tecentriq), durvalumab, dacomitinib, osimertinib,
tivantinib,
onartuzumab, sorafenib, crizotinib, ceritinib, trametinib, dabrafenib,
bevacizumab (avastin),
exisulind, bexarotene, cetuximab, squalamine, necitumumab, cyclophosphamide,
ramucirumab, doxorubicin, porfimer, methotrexate, an FDA and/or EMA approved
drug(s) or
treatment(s) for lung cancer (including, without limitation, radiofrequency
ablation,
microwave ablation, cryoablation, thermal ablation, chemoembolization),
optionally with
plasma [almitrine] and/or [lactate] and/or [bilirubin] recording(s) before
and/or during the
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CA 3050553 2019-07-25
course of almitrine administration, wherein if these are abnormally/too high,
almitrine
administration is reduced/stopped/not commenced. Almitrine greatly assists the
treatment/amelioration/prevention/combat of NSCLC, which can be very radio-
(e.g. refer
radioresistance of NCI-H460 cell line in [319]) and chemo- resistant, wherein
almitrine
undermines this resistance, enabling greater radio- and chemo- therapy to
occur, improving
the clinical outcome of the subject and/or their quality of life. It
especially, without =
limitation, helps subjects with NSCLC that can't/won't undergo surgery, or
whose cancer is
inoperable, and who must completely rely on radio- and/or chemo- therapy for
cancer
treatment, wherein some NSCLC cancers are very radio- [319] and chemo-
resistant and
thence incredibly dangerous.
Almitrine is mentioned, in combination with a plant extracted compound, in the
Simplified
Chinese to English machine translation of CN105693806A [P13]. The "First
Office Action"
from The State Intellectual Property Office of People's Republic of China for
CN105693806A identifies that CN105693806A is not a teaching document because
it
presents exactly the same extraction protocol as another Chinese filing
(CN201610258319)
but wherein a very different compound is extracted, which is highly
improbable, to the point
of incredulity, as reported by the Chinese Examiner. Furthermore the anti-
cancer data points
(and even error bars) in CN105693806A are identical to that of CN201610258319,
despite
different compounds being tested, which is an extremely improbable result,
again, to the
point of incredulity, again, as identified by the Chinese Examiner, wherein
CN105693806A
and CN201610258319 were both filed on the same day (23/04/2016) by different
inventors.
Furthermore, perversely, there are many other Chinese patent applications,
from same and
different inventors, many submitted on the same or within days of one another,
that report the
exact same protocol as CN105693806A for extracting very different compounds,
which is
extremely strange and improbable, again to the point of incredulity, as
highlighted by the
Chinese examiner of CN105693806A in their First Office Action report, wherein
some of
these other applications are listed. But there are many further such
applications findable by
one of the art (>100), and their examination reports typically identify
fabrication in each case,
while further identifying that this fabrication is componentry to a wider
fabrication of
multiple patent filings, with shared characterizing features (e.g. same
extraction protocol used
in every case). So there are many legal documents teaching one of the art
about this iterated
fabrication scheme, of which CN105693806A is apart. Refer, for non-limiting
example, to
the following filings and (especially) their Chinese Intellectual Property
Office examination
391
CA 3050553 2019-07-25
reports: CN201610111060 (CN105622711), CN201610157201 (CN105585609),
CN201610167771 (CN105663143A), CN201610167879 (CN105837393A),
CN201610168492 (CN I05777682A), CN201610172606 (CN105646642), CN201610203075
(CN105837656), CN201610263608 (CN 1 05884773A), CN201610331470
(CN I06008539A), CN201610331596 (CN106008542A), CN201610333811
(CN105906647A), CN201610334869 (CN105924339A), CN201610335078
(CN I 05859535A), CN201610335171 (CN105884792A), CN201610335324
(CN105906683A), CN201610338448 (CN105924413A), CN201610340391
(CN105949216A), CN201610343221 (CN 1 06008248A), CN201610473707
(CN106083988A), CN201610493013 (CN106046112A), CN201610580967
(CN106220702A). So, to recap, the Chinese examiner of CN105693806A concluded
that
CN105693806A is not a teaching document. CN105693806A is, or at least would be
surmised by one of the art, to be a fabricated filing. There are motivations
for fabricating a
patent filing(s), for example in China, some of which are disclosed in this
Economist
magazine article [320]. In China "workers and students who file patents are
more likely to
earn a hukou (residence permit) to live in a desirable city", "for some
patents the government
pays cash bonuses", "corporate income tax can be cut from 25% to 15% for firms
that file
many patents" and "they are also more likely to win lucrative government
contracts", "many
companies therefore offer incentives to their employees to come up with
patentable ideas",
"the generosity of China's incentives for patent-filing may make it worthwhile
for companies
and individuals to patent even worthless ideas", "most of these patents are
probably filed with
the expectation that they will be ignored" [320]. Refer also to this Bloomberg
article [321]. In
short, the official Chinese Intellectual Property Office examination report
(First Office
Action) of CN105693806A, reporting that CN105693806A is not a teaching
document,
teaches away from CN105693806A.
EXAMPLE (VII)
Formula (VII): Any amino acid sequence/protein/peptide/polypeptide/antibody,
either
naturally occurring or produced by engineering or fortuity, that
preferentially/disproportionally/selectively inhibits the "reverse" ATP
hydrolysing mode as
compared to the "forward" ATP synthesising mode of ATP synthase. Note that
selectively
inhibiting FIFO ATP hydrolysis can seemingly inhibit FiFo ATP synthesis
because less ATP is
made, but this is because less ATP is hydrolysed and so less ATP needs to be
made, rather
than any actual direction inhibition upon FIFO ATP synthesis. Preferred
embodiments are
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CA 3050553 2019-07-25
those that inhibit the reverse mode of ATP synthase potently and the forward
mode of ATP
synthase less potently, and most preferably not at all. If FiFo ATP synthesis
falls because of
inhibited FiFo ATP hydrolysis, and not primarily because of direct inhibition
of FIFO ATP
synthesis, then this compound is still within the scope of this invention.
Componentry to Formula (VII) is one or more of melittin, the presequence of
yeast
cytochrome oxidase subunit IV and each synthetic derivative of this
presequence ([4],
incorporated in its entirety). Also componentry to Formula (VII) is one or
more IF1 proteins
from any eukaryote, and/or one or more of any IF1 subsequence e.g. any minimal
inhibitory
IF I sequence, which, for example, for bovine IF I is amino acid residues 14
to 47 [330, 331],
and/or any IF1 protein variant especially wherein this variant has greater
inhibitory potency
against FIFO ATP hydrolysis, and/or one or more of any gene or nucleotide or
DNA or RNA
that translates to one or more of any IF1 protein, IF1 protein variant, IF1
subsequence and
IF1 subsequence variant. These examples just given, and further examples
herein, for
Formula (VII) are illustrative and not restrictive.
IF1 proteins are highly conserved and typically interchangeable between
species [330]. An
invention embodiment is an increased amount of IF1 protein in a subject,
beyond normally
occurring IF1 levels in a subject, and/or expression of an IF1 protein from a
different species
and/or an IF1 protein variant(s), and/or subsequence variant(s), most
preferably one that has
greater inhibitory action against FIR) ATP hydrolysis at normal matrix pH (-8)
e.g. the H49K
(mature {mitochondrial import signal sequence cleaved off} IF1 protein
numbering) mutant
of bovine IF1 [330, 332], wherein methods of introducing genes and/or gene
copies and/or
DNA and/or RNA and/or proteins into a subject are well known to those of the
art e.g. refer
[322], which expresses H49K mutant human [Fl in mice (this amino acid
substitution makes
IF1 constitutively active at normal matrix pH {-8}). An invention embodiment
is bovine IF I
expressed in a subject, wherein this IF1 has one or more of its histidine
residues at positions
His-48, 49, 55 changed to another amino acid, for example alanine or lysine.
In another
embodiment, IF1 from a non-bovine species is expressed in a subject, wherein
this 1F1 is
changed at the equivalent histidine positions to those aforementioned in the
bovine IF I
sequence (highly conserved across species, but not always with same amino acid
numbering),
which renders the IF1 with greater inhibitory potency against FIFO ATP
hydrolysis at normal
matrix pH (-8) [330, 332]. A further invention embodiment is the expression of
IF1 in a
subject (a native IF1 sequence to a species or an altered [Fl sequence herein
described) that
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CA 3050553 2019-07-25
has one or more amino acid changes that don't significantly alter its
inhibitory potency
against FIF0 ATP hydrolysis (could be termed neutral changes), and more
preferably that
increases its inhibitory potency against FiFo ATP hydrolysis.
Larger species have more IFI [330], and/or use an IF I variant that has
greater inhibitory
potency for FIR' ATP hydrolysis, a lower specific metabolic rate, less
metabolic heat
production per unit mass, and longer lifespan (Figure 24). Increasing the
amount of IF1,
and/or expressing an IF I variant that has greater inhibitory potency for FiFo
ATP hydrolysis,
decreases metabolic rate and extends lifespan, so long as exogenous heat
substitutes for the
lesser endogenous heat production, or greater body insulation (e.g. wearing
more clothes)
renders same body temperature for a lessor metabolic rate. An invention
embodiment is to
use an animal(s), whose quantity and/or nature of its IF1 is manipulated, in a
lifespan and/or
healthspan study, optionally entering it into a competition with a financial
or other reward
e.g. the M prize. To extend the lifespan of a subject, for example a mouse or
human, increase
the amount of its own IF1 sequence and/or express one or more of the IF1
sequence of a
bigger and/or longer living species. The naked mole rat (Heterocephalus
glaber) expresses
ATPIF I five times more than the mouse [342], and lives ten times longer. An
invention
embodiment is to express the IF1 (National Center for Biotechnology
Information [NCBI]
Gene ID: 101712500), and/or its further IF1 like gene (NCBI Gene ID:
110349814), of the
naked mole rat in a homeothermic species to slow its metabolism and extend its
lifespan, with
the caveat that this species will now acquire some or all of the
thermoconformer character of
the naked mole rat and now must be kept at higher ambient temperature: naked
mole rats can
survive without homeothermy because they live in hot East African countries,
permanently
underground in burrows (wherein heat is maintained at night), even eating
underground
(tubers), and can behaviourally thermoregulate by choosing their depth in the
burrow.
Any method wherein a compound(s), including protein and/or antibody compounds,
that
inhibits FIFO ATP hydrolysis is administered to a subject to increase/improve
the subject's
health, healthspan and/or lifespan, and/or to reduce/slow the
signs/damage/maladies/diseases
of aging, is componentry to the present invention.
For reference, human IF1 amino acid sequence, with pre-sequence (using one
letter amino
acid code; NCBI Gene ID: 93974):
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MAVTALAARTWLGVWGVRTMQARGFGSDQSENVDRGAGSIREAGGAFGKREQAE
EERYFRAQSREQLAALKKHHEEEIVHHKKEIERLQKEIERHKQKIKMLKHDD
Human IF1 amino acid sequence, with less pH dependency (has a histidine
changed to
another amino acid, in this case a lysine):
MAVTALAARTWLGVWGVRTMQARGFGSDQSENVDRGAGSIREAGGAFGKREQAE
EERYFRAQSREQLAALKKHKEEEIVHHKKEIERLQKEIERHKQKIKMLKHDD
Another human IF1 amino acid sequence, with less pH dependency (key histidines
changed
to another amino acid, alanine in each case here to illustrate):
MAVTALAARTWLGVWGVRTMQARGFGSDQSENVDRGAGSIREAGGAFGKREQAE
EERYFRAQSREQLAALKKAAEEEIVAHKKEIERLQKEIERHKQKIKMLKHDD
Naked mole rat IF1 sequence (NCBI Gene ID: 101712500):
MAGTALASRARLGVWGVRAMQTRGFSSDKDHESSSGSIRDAGGAFGKREQAEEERF
FRQKTKEQLEALKKHHEDEIYHHKKAIEHMEKEIERHKQKIKQLKHDD
Naked mole rat IF1 like sequence (NCBI Gene ID: 110349814):
QDHESSSGSIRDAGGAFGKREQAEEEPFFQQKTKEQLEALKKHHEDEIYHHKKAIEC
MEKEIERHKQKIKQLKHDD
Melittin sequence: GIGAVLKVLTTGLPALISWIKRKRQQ-NH2
Presequence of yeast cytochrome oxidase subunit IV:
MLSLRQSIRFFKPATRTLCSSRYLL-NH2
Synthetic derivatives of yeast cytochrome oxidase subunit IV presequence:
MLSLRQSIRFPATRTLCSSRYLL-NH2
MLSRLSLRLLSRLSLRLLSRYLL-NH2
MLSSLLRLRSLSLLRLRLSRYLL-NH2
Some protein sequence embodiments of the invention are presented in Figure 29
and in the
SEQUENCE LISTING componentry to this disclosure.
Cyclic peptide embodiments of the invention
Componentry to this invention is any IF1 sub-sequence(s)/FIFo ATP hydrolysis
inhibitory
peptide(s), or mutant thereof, optionally with one or more non-proteingenic
(e.g. D-) amino
acids (increases plasma stability), optionally lipidated (fatty acid(s)
attached), optionally with
one or more Na-methylated, optionally with a cell penetrating peptide (CPP)
sequence(s), in a
cyclic form (optionally monocyclic, bicyclic, tricyclic, or higher cycle
number), wherein a
CPP sequence(s) for cyclic forms is preferred e.g. (non-limiting) as disclosed
in
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US20170190743A1, US20170355730A1, W02015/179691A2, [349-350]. In a bicyclic
form,
in some invention embodiments, an IF1 sub-sequence(s)/Fi Fo ATP hydrolysis
inhibitory
peptide(s) sequence is in one cycle and a CPP sequence(s) in the other cycle.
So, the target
binding and CPP sequences are in different cycles of a bicyclic form, wherein,
to illustrate
(not restrict), this division (for a different target) was used in [351]. In
some embodiments, an
IF1 sub-sequence(s)/FIF0 ATP hydrolysis inhibitory peptide(s) sequence, with a
CPP
sequence, is incorporated into a reversible bicycle structure which becomes
linear in the
reducing intracellular environment, to illustrate (not restrict), as used in
[352], wherein 3,5-
bis(mercaptomethyl)benzoyl [BMB] and two cysteine residues are incorporated
into the
sequence judiciously, optionally with sequence order: BMB ¨ CPP ¨ cysteine ¨
[IF1 sub-
sequence] ¨ cysteine. In alternative embodiments, the CPP is in an
intracellular labile cycle
and the IF1 sub-sequence is linear, optionally implemented by using 3-
mercaptopropionyl
(MP) and sequence order: MP ¨ CPP ¨ cysteine ¨ [IF1 sub-sequence], following
teaching of
[352]. In some embodiments, an IF1 sub-sequence(s)/FIF0 ATP hydrolysis
inhibitory
peptide(s) sequence, or mutant thereof, optionally with one or more non-
proteingenic amino
acids, is incorporated into a bicycle structure, optionally with the same
sequence in both
cycles, optionally with a different IF1 sub-sequence/FIF0 ATP hydrolysis
inhibitory sequence
in each cycle, optionally with a CPP sequence also in both or just one of the
cycles,
optionally wherein a CPP sequence occupies one of the cycles and an IF1 sub-
sequence(s)/FiFo ATP hydrolysis inhibitory sequence(s) occupies the other
cycle, optionally
wherein the bicycle structure is made according to a teaching of the art e.g.
[353-358] , e.g. as
exemplified by the publication/patent/drug output of Bicycle Therapeutics
Ltd., (Babraham
Research Campus, Cambridge, U.K) and/or its
subsidiaries/affiliates/founders/investors/acquirer(s).
An IF! sub-sequence(s)/FIF0 ATP hydrolysis inhibitory peptide(s), or mutant
thereof,
optionally with a cell penetrating peptide (CPP) sequence(s), in a cyclic form
(optionally
monocyclic, bicyclic, tricyclic, or higher cycle number) is componentry to
this invention.
Some non-limiting invention embodiments, disclosed using one-letter amino acid
code and
further symbols designated herein: Cyclo(F(DRRRRx); Cyclo(xFORRRR);
Cyclo(RRFRORx); Cyclo(xRRFROR); Cyclo(FORRRx); Cyclo(xFORRR);
Cyclo(FORRRRRx); Cyclo(xFORRRRR); Cyclo(FFORRRRx); Cyclo(xFFORRRR);
Cyclo(RFRFRORx); Cyclo(xRFRFROR); Cyclo(fORrRrx); Cyclo(xfORrRr);
Cyclo(fearRrRx); Cyclo(xfORrRrR); Cyclo(FfelltrRrx); Cyclo(xFfORrRr);
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Cyclo(F9rRrRx); Cyclo(xF(prRrR); Cyclo(FWRRRRx); Cyclo(xFWRRRR);
Cyclo(YORRRRx); Cyclo(xYORRRR); Cyclo(gORRRRx); Cyclo(xgORRRR);
Cyclo(F9rRrRx); Cyclo(xF9rRrR);
wherein,O= L-2-naphthylalanine, 9 = D-2-naphthylalanine, f= D-phenylalanine, r
= D-
arginine, g = L-phenylglycine, x = IF1 sub-sequence(s)/F 1F0 ATP hydrolysis
inhibitory
peptide(s) {in either orientation}, optionally one of following sequences (or
one of these
reversed): H, HH, HHH, HHHH, HHHHH, HHHHHH, HHHHHHH, HHHHHHHH, HHE,
HHEE, HHEEE, HHEEE1, HHEEEIV, HHEEEIVH, HHEEEIVHH.
In further embodiments,
(ANyN /(AA)r, (AA)v (AA) y z(NA)n (AA)m (AA)v
Cys Cys Cys Cys Cys
S--I Ls
Loop 1 z3 Loop 2
zi Monocyclic z2 zi bicyclic z2
AA = amino acid (proteingenic or non-proteingenic);
Cys = cysteine;
y and v are independently selected from 0 and an integer between 1-100;
n and m are independently selected from an integer between 1-20;
zl, z2, z3 are independently selected from an integer between 1-5;
(AA)n = IF1 sub-sequence(s)/FIF0 ATP hydrolysis inhibitory
peptide(s)/histidine(s);
(AA)m = IF1 sub-sequence(s)/FiFo ATP hydrolysis inhibitory peptide(s)/cell
penetrating
peptide (CPP) sequence;
Optionally, (AA)m is selected from one of FORRRR, RRRROF, RRFROR, RORFRR,
=
FORRR, RRROF, FORRRRR, RRRRROF, FFORRRR, RRRROFF, RFRFROR,
RORFRFR, fORrRr, rRritil)f, filarRrR, RrRrRilf, FfetRrRr, rRrRoDfF, F9rRIR,
RrRr9F,
FWRRRR, RRRRWF, YORRRR, RRRROY, gilaRRR, RRRROg, F9rRrR, RrRr9F;
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Optionally, (AA)n is selected from one of H, HH, HHH, HHHH, HHHI-IH, HHHHHH,
HHHHHHH, HHHHHHHH, HHE, HHEE, HHEEE, HHEEE1, HHEEEIV, HHEEEIVH,
HHEEEIVHH.
Gene therapy embodiments of the invention
Componentry to this invention are the nucleic acid sequences that code for the
protein
sequences of the invention. In a specific embodiment, a nucleic acid sequence
encoding one
or more protein embodiments of the invention is administered to a subject,
optionally to treat
and/or enhance the subject, by way of gene therapy. Any method for gene
therapy available
in the art can be used according to the present invention. For general reviews
of the methods
of gene therapy, refer Goldspiel et al., 1993, Clinical Pharmacy 12:488-505;
Wu and Wu,
1991, Biotherapy 3:87-95; Tolstoshev, 1993, Ann. Rev. Pharmacol. Toxicol.
32:573-596;
Mulligan, Science 260:926-932 (1993); and Morgan and Anderson, 1993, Ann. Rev.
Biochem. 62:191-217; May, 1993, TIBTECH 11(5): 155-215. Methods known in the
art of
recombinant DNA technology can be used, some of which are described in Ausubel
et al.
(eds.), Current Protocols in Molecular Biology, John Wiley & Sons, NY (1993);
and
Kriegler, Gene Transfer and Expression, A Laboratory Manual, Stockton Press,
NY (1990).
In a preferred aspect, a composition of the invention comprises a nucleic acid
sequence,
encoding a protein sequence embodiment of the invention, in an expression
vector that
expresses the nucleic acid sequence into a protein sequence of the invention.
Such a nucleic
acid sequence has a promoter, optionally a heterologous promoter, operably
linked to the
protein coding region. Said promoter is inducible or constitutive, and,
optionally, tissue-
specific. With an inducible promoter, expression of the nucleic acid is
controllable by
controlling the presence or absence of the appropriate inducer of
transcription (non-limiting
.. e.g. "Tet-Off' {transcription is inactive in presence of tetracycline or
doxycycline or similar}
and "Tet-On" {transcription is only active in presence of tetracycline or
doxycycline or
similar} expression systems). In a particular embodiment, the nucleic acid
sequence used has
the sequence coding for the therapeutic protein embodiment, and optionally its
regulatory
region(s) {promoter(s) etc.}, flanked by regions that promote homologous
recombination at a
desired site in the genome, thus providing for intrachromosomal expression of
the protein
encoding nucleic acid sequence (Koller and Smithies, 1372718-2 72 1989, Proc.
Natl. Acad.
Sci. USA 86:8932-8935; Zijlstra et al., 1989, Nature 342:435 438). Delivery of
the nucleic
acid into a subject may be direct, by directly exposing the subject to the
nucleic acid or
nucleic acid-carrying vector, or indirect, wherein cells are transformed with
the nucleic acid
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in vitro, then transplanted into the subject: termed in vivo and ex vivo gene
therapy
respectively. In a specific embodiment, a nucleic acid sequence of this
invention is
incorporated into a nucleic acid expression vector and administered to the
subject such that it
becomes intracellular e.g. by infection using a defective or attenuated
retroviral or other viral
vector (see U.S. Pat. No. 4,980.286), or by direct injection of naked DNA, or
by use of
microparticle bombardment (e.g., a gene gun; Biolistic, Dupont), or by a
matrix with in situ
scaffolding in which the nucleic acid Sequence is contained (See, e.g.,
European Patent No.
EP 0 741 785 BI and U.S. Pat. No. 5,962,427), or coating with lipids or cell-
surface receptors
or transfecting agents, encapsulation in liposomes, microparticles, or
microcapsules, or by
administering in linkage to a peptide which is known to enter the nucleus, by
administering it
in linkage to a ligand subject to receptor-mediated endocytosis (see, e.g., Wu
and Wu, 1987,
J. Biol. Chem. 262:4429-4432) (which can be used to target cell types
specifically expressing
the receptors), etc. In another embodiment, nucleic acid-ligand complexes can
be formed in
which the ligand comprises a fusogenic viral peptide to disrupt endosomes,
allowing the
nucleic acid to avoid lysosomal degradation. In yet another embodiment, the
nucleic acid can
be targeted in vivo for cell specific uptake and expression, by targeting a
specific receptor
(See, e.g., PCT Publication Nos. WO 92/06180, WO 92/22635, WO 92/20316, WO
93/14188, and WO 93/20221). Alternatively, the nucleic acid can be introduced
intracellularly and incorporated within host cell DNA for expression, by
homologous
recombination (Koller and Smithies, 1989, Proc. Nat). Acad. Sci. USA 86:8932-
8935; and
Zijlstra et al., 1989, Nature 342:435-438). In a specific embodiment, a viral
vector that
contains a nucleic acid sequence of this invention is used. For example, a
retroviral vector
can be used (see, e.g., Miller etal., 1993, Meth. Enzymol. 217:581 599). These
retroviral
vectors contain the components necessary for the correct packaging of the
viral genome and
integration into the host cell DNA. More detail about retroviral vectors can
be found in
Boesen et al., 1994, Biotherapy 6:291-302, which describes the use of a
retroviral vector to
deliver the MDR I gene to hematopoietic stem cells in order to make the stem
cells more
resistant to chemotherapy. Other references illustrating the use of retroviral
vectors in gene
therapy are: Clowes et al., 1994, J. Clin. Invest. 93:644-651; Klein et al.,
1994, Blood
83:1467-1473; Salmons and Gunzberg, 1993, Human Gene Therapy 4:129-141; and
Grossman and Wilson, 1993, Curr. Opin. in Genetics and Devel. 3:110-114.
Adenoviruses
are other viral vectors that can be used in gene therapy. Adenoviruses are
especially attractive
vehicles for delivering genes to respiratory epithelia. Adenoviruses naturally
infect
respiratory epithelia where they cause a mild disease. Other targets for
adenovirus-based
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delivery systems are liver, the central nervous system, endothelial cells, and
muscle.
Adenoviruses have the advantage of being capable of infecting non-dividing
cells. Kozarsky
and Wilson, 1993, Current Opinion in Genetics and Development 3:499-503
present a review
of adenovirus based gene therapy. Bout et al., 1994, Human Gene Therapy 5:3-10
demonstrated the use of adenovirus vectors to transfer genes to the
respiratory epithelia of
rhesus monkeys. Other instances of the use of adenoviruses in gene therapy can
be found in
Rosenfeld et al., 1991, Science 252:431-434; Rosenfeld et al., 1992, Cell
68:143-155;
Mastrangeli et al., 1993, J. Clin. Invest. 91:225-234, PCT Publication
W094/12649; and
Wang et al., 1995, Gene Therapy 2:775-783. Adeno-associated virus (AAV) is
another option
for use in gene therapy (Walsh et al., 1993, Proc. Soc. Exp. Biol. Med.
204:289-300; and U.S.
Pat. No. 5,436,146). Another option is to use an ancestral AAV such as Anc80
(Zinn et al.
2015, Cell Reports, 12(6):1056-68). Another approach to gene therapy involves
transferring a
gene to cells in vitro, in tissue culture, by such methods as electroporation,
lipofection,
calcium phosphate mediated transfection, or viral infection. Usually, the
method of transfer
includes the transfer of a selectable marker to the cells. The cells are then
placed under
selection to isolate those cells that have taken up and are expressing the
transferred gene.
Those cells are then delivered to a subject. In an embodiment, the nucleic
acid is introduced
into a cell prior to administration in vivo of the resulting recombinant cell.
Such introduction
can be carried out by any method known in the art, including but not limited
to transfection,
electroporation, microinjection, infection with a viral or bacteriophage
vector containing the
nucleic acid sequence(s), cell fusion, chromosome-mediated gene transfer,
microcellmediated
gene transfer, spheroplast 1372718-2 74 fusion, etc. Numerous techniques are
known in the
art for the introduction of foreign genes into cells (see, e.g., Loeffler and
Behr, 1993, Meth.
Enzymol. 217:599-618; Cohen et al., 1993, Meth. Enzymol. 217:618-644; Clin.
Pharma.
Ther. 29:69-92 (1985)) and may be used in accordance with the present
invention. The
technique should provide for the stable transfer of the nucleic acid to the
cell, so that the
nucleic acid is expressible by the cell and, optionally, heritable and
expressible by its cell
progeny. The resulting recombinant cells can be delivered to a subject by
various methods
known in the art. Recombinant blood cells (e.g., hematopoietic stem or
progenitor cells) are
preferably administered intravenously. The amount of cells envisioned for use
depends on the
desired effect, patient state, etc., and can be determined by one skilled in
the art. Cells into
which a nucleic acid can be introduced for purposes of gene therapy encompass
any desired,
available cell type, and include but are not limited to epithelial cells,
endothelial cells,
keratinocytes, fibroblasts, muscle cells, hepatocytes, blood cells Such as T
lymphocytes, B
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lymphocytes, natural killer (NK) cells, monocytes, macrophages, neutrophils,
eosinophils,
megakaryocytes, granulocytes, various stem or progenitor cells, in particular
hematopoietic
stem or progenitor cells, e.g., as obtained from bone marrow, umbilical cord
blood, peripheral
blood, fetel liver, etc. In a preferred embodiment, the cell used for gene
therapy is autologous
to the subject. In a specific embodiment, stem or progenitor cells are used.
Any stem and/or
progenitor cells which can be isolated and maintained in vitro can potentially
be used in
accordance with this embodiment of the present invention (see e.g., PCT
Publication No.
W094/08598; Stemple and Anderson, 1992, Cell 7 1:973-985; Rheinwald, 1980,
Meth. Cell
Bio. 21A:229; and Pittelkow and Scott, 1986, Mayo Clinic Proc. 61:771).
"Tet-Off" and "Tet-On" inducible gene expression embodiments of the invention
In a "Tet-Off' system, gene expression is under the control of a tetracycline-
responsive
promoter element (TRE), which permits gene expression if the tetracycline-
controlled
transactivator protein (tTA) binds, wherein tTA binding is blocked by the
presence of
tetracycline or doxycycline or similar compound structure. In a "Tet-On"
system, gene
expression is under the control of a tetracycline-responsive promoter element
(TRE), which
permits gene expression if the reverse tetracycline-controlled transactivator
(rtTA) binds,
wherein rtTA binding is enabled and conditional upon the presence of
tetracycline or
doxycycline or similar compound structure. So, one can transfer a gene into an
organism
under the control of a TRE promoter, and then control this gene's expression,
spatially and/or
temporally, by (1) controlling whether tTA or rtTA is expressed (which is set
by which of
these genes is also gene transferred into the organism) and (2) whether a
tetracycline type
compound structure is present or absent (set by compound administration or
lack thereof).
For example, [322] generated transgenic mice with a human IF1 mutant gene (a
histidine in
the "pH dependence motif' {refer Figure 29} substituted with lysine) under a
TRE promotor.
Furthermore, these mice had another exogenous gene present, for tTA protein
expression, but
only expressed in its forebrain neurons, and thence the mutant human IF I gene
was only
expressed in forebrain neurons (wherein this expression is dependent on
absence of
tetracycline or similar structure). [322] doesn't specify the ambient
temperature these mice
.. were housed at. With this omission, I presume normal room temperature,
which is typically
in the range 20-25 C. [322] is an illustrative example of how an IF1 gene
copy, or a mutant
thereof, from the same or different species, can be transferred into an
organism in order to
increase its IF1 protein expression. This example shows it is safe in mouse
brain (more
specifically neurons in forebrain [323], [322] is incorrect to refer to whole
brain as it does) to
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CA 3050553 2019-07-25
increase IF1 protein content by three times, wherein the delta increase in
[IPI] occurs with a
mutant human IF1 form with increased inhibitory potency against FIF0 ATP
hydrolysis at pH
8, which reduces FIFO ATP hydrolysis capability by ¨35%, which demonstrates
the safety of
inhibiting FIFO ATP hydrolysis in vivo, at least specifically in forebrain
neurons (mice were
"normal in appearance, home-cage behavior, reproduction, and longevity up to 1-
year follow-
up"). For more data and analysis on this herein, refer Figure 25 and legend,
and then Figures
26 and 27 and their legends. In alternative embodiments, tTA is expressed in
all the tissues of
the mouse, not just the brain (or part(s) thereof), and this mouse, and others
like it, are
entered into a lifespan study. These mice will have a longer health- and life
¨span than
control mice (control mice optionally have the transgene for tTA but not the
transgene for IF1
or IF I mutant, or are the same genetically but are administered doxycycline,
or other
tetracycline, so that they don't express the extra IF1 gene), especially if
the mice are kept at
37 C.
Further "Tet-Off' embodiments of the invention
An invention embodiment is for an IF1 protein coding gene (herein referred to
as an IF1
gene), optionally a mutant thereof, optionally with a nucleotide change(s)
that causes an
amino acid residue change(s) at the "phosphorylation control switch" and/or
the "pH
dependence motif' in the IF! protein coded for (these IF I elements are
defined in Figure 29),
to be expressed only, or only substantially, or disproportionally, or more, in
a specific cell
type/tissue/organ/area of a subject, optionally wherein this selective IF1
expression reduces
Fi Fo ATP hydrolysis and slows the metabolic/aging rate in this sub-section(s)
of the subject,
wherein this sub-section(s) generates less metabolic heat, but this is
substituted for by heat
transfer from surrounding body areas. For illustrative example, by making a
transgenic mouse
that expresses tTA specifically in a certain mouse body tissue, by putting tTA
expression
under the control of a tissue specific promoter, wherein this tissue specific
expression of tTA
then drives tissue specific expression of an introduced IF1 gene, optionally a
mutant thereof,
whose expression is under the control of a tetracycline-responsive promoter
element (TRE).
For non-limiting example, putting tTA expression under the control of a
dopamine neuron
specific promoter such as that for tyrosine hydroxylase (the first and rate-
limiting enzyme for
dopamine synthesis) [363], or the dopamine transporter (DAT, required for
dopamine re-
uptake into dopaminergic neurons) [364, 365], or Pitx3 (transcription factor
involved in
dopaminergic neuron differentiation), or DIA dopamine receptor subtype, which
then drives
specific expression of an IF I gene, or mutant thereof, in dopaminergic
neurons, wherein this
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CA 3050553 2019-07-25
IF1 gene is under the control of a tetracycline-responsive promoter element
(TRE). Greater
IF1 protein expression in dopaminergic neurons reduces their F IF ATP
hydrolysis,
especially if the extra IF1 protein has an amino acid residue that can't be
phosphorylated
(optionally alanine) at its "phosphorylation control switch" position and/or a
H49K
substitution (position 49 using mature {mitochondria' import signal sequence
cleaved oft)
human IF1 protein numbering) in its "pH dependence motif', which slows the
metabolic/aging rate in dopaminergic neurons, which reduces the subject's
risk/progression
of Parkinson's disease, wherein these dopaminergic neurons generate less
metabolic heat, but
this is substituted for by heat transfer from surrounding body areas, such
that the
.. dopaminergic neurons are maintained at normal body temperature. A useful
control is that
this subject, when administered (e.g. in drinking water) with tetracycline,
has the same
risk/progression of Parkinson's disease as control subject without these
genetic manipulations
(which are, to recap, addition of tTA gene, under dopaminergic neuron specific
promoter, and
addition of IF1 gene, or mutant thereof, under control of a tetracycline-
responsive promoter
element, TRE). In another invention embodiment, tTA expression is put under
the control of
a photoreceptor specific promoter, such as that for rhodopsin (or other
opsin), which then
drives specific expression of an IF1 gene, or mutant thereof, in
photoreceptors, wherein this
IF1 gene is under the control of a tetracycline-responsive promoter element
(TRE). Greater
IF1 protein expression in photoreceptors reduces their FIFO ATP hydrolysis,
especially if the
extra IF1 protein has an amino acid residue that can't be phosphorylated
(optionally alanine)
at its "phosphorylation control switch" position and/or a H49K substitution
(position 49 using
mature {mitochondria' import signal sequence cleaved off) human IF I protein
numbering) in
its "pH dependence motif', which slows the metabolic/aging rate in
photoreceptors, which
reduces the subject's risk/progression of aging related/correlated eye
disease(s), optionally
age-related macular degeneration (AMD), wherein these photoreceptors generate
less
metabolic heat, but this is substituted for by heat transfer from surrounding
body areas, such
that the photoreceptors are maintained at normal body temperature.
In an invention embodiment, the experiment of [322] is repeated but with the
mutant IF1
transgene expressed in a different part of the mouse. So, instead of using B16-
Tg(Camk2a-
tTa)1Mmay/J mice, which express tTA in forebrain neurons, as [322] used, a
different
transgenic mouse type can be used, which expresses tTA in a different part of
the mouse
body, which drives mutant IF1 expression in that different part of the mouse
body.
Illustratively, a variety of transgenic mice that express tTA in different
body
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areas/tissues/organs are available to those of the art, e.g. refer to a
database of such mice at
[366], and a number are commercially available, for illustrative example from
The Jackson
Laboratory (USA). Wherein (stock numbers given are for The Jackson
Laboratory), for non-
limiting example, B6.Cg-Tg(GFAP-tTA)110Pop/.1 mouse (stock No: 005964) has tTA
expression driven by the human glial fibrillary acidic protein (GFAP) promoter
and expresses
tTA in astrocytes, B6.Cg-Tg(Sirpa-tTA)AUmri/J mouse (stock No: 023970) has tTA
expression driven by mouse signal-regulatory protein alpha (Sirpa) promoter
and expresses
tTA in dentate granule cells of the hippocampus, B6.Cg-Tg(Scg2-tTA)1Jta mouse
(stock No:
008284) has tTA expression driven by mouse secretogranin II promoter and
expresses tTA in
the brain, especially the suprachiasmatic nucleus, B6.Cg-Tg(Eno2tTA)5030Nes/J
mouse
(stock No: 003763) has tTA expression driven by rat neuron-specific enolase
(Eno2)
promoter and expresses tTA at high levels in the striatum and to a lesser
extent in the cerebral
cortex and hippocampus, B6;C3-Tg(NEFH-tTA)8V1e/J mouse (stock No: 025397) has
tTA
expression driven by human neurofilament heavy polypeptide (NEFH) promoter and
expresses tTA in neurons and large-caliber axons of the brain and spinal cord,
B6;129S-
51c6a3tm4.1(tTA)Xza mouse (stock No: 027178) has tTA expression driven by
mouse
Slc6a3 (solute carrier family 6, neurotransmitter transporter, dopamine, DAT)
promoter and
expresses tTA in dopamine neurons, NOD.Cg-Tg(Ins2-tTA)1Doi/DoiJ mouse (stock
No:
004937) has tTA expression driven by rat insulin promoter (Ins2, commonly
designated RIP)
and expresses tTA in pancreatic beta cells, FVB.Cg-Tg(Myh6-tTA)6Smbf/J mouse
(stock
No: 003170) has tTA driven by rat alpha myosin heavy chain promoter and
expresses tTA in
cardiac myocytes. Many other such mice are commercially available, and/or are
known to
those of the art (e.g. from the literature) and other such mice, with tTA
(and/or rtTA)
specifically expressed in other cell type(s)/tissue(s)/organ(s), can be
generated using
techniques of the art, wherein how to apply these techniques to other mammal
species is
known to those of the art.
An invention embodiment is for an IF1 gene, optionally a mutant thereof,
optionally with a
nucleotide change(s) that causes an amino acid residue change(s) at the
"phosphorylation
control switch" and/or the "pH dependence motif' (these IF1 elements are
defined in Figure
29) in the IF1 protein coded for, to be expressed ubiquitously in a subject,
in every tissue. For
illustrative example, by making a transgenic mouse that expresses tTA
ubiquitously, by
putting tTA expression under the control of a ubiquitous (e.g. promoter for
actin gene) or
synthetic (e.g. CAG promoter [367]) promoter, wherein this ubiquitous
expression of tTA can
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then drive ubiquitous expression of an introduced IF! gene, optionally a
mutant thereof,
whose expression is under the control of a tetracycline-responsive promoter
element (TRE).
More direct IF1 genetic manipulations of this invention
An invention embodiment is to render localized or ubiquitous expression of an
extra IFI
gene(s) in a subject, optionally a mutant thereof, directly, without using tTA
or rtTA and
TRE. So, to illustrate, and not restrict, to render tissue specific IF1
expression in a subject by
administering the IF1 gene(s), or mutant thereof, locally (rather than
systemically) and/or
under the control of a tissue specific promoter or, alternatively, rendering
ubiquitous IF!
expression in a subject by introducing the IF1 gene(s), or mutant thereof,
under the control of
a ubiquitous promoter, optionally a synthetic promoter. Without restriction,
there are two
possible options: (1) the introduced IF1 gene(s), or mutant thereof, under the
control of an
introduced promoter (and/or enhancer and/or other regulatory element(s)), is
introduced to
the subject or (2) the IF1 gene(s) only, or mutant thereof, is introduced to
the subject, wherein
it inserts into the genome at a place that renders its expression under the
control of a
cell/tissue/organ specific or ubiquitous promoter already in the genome. The
IF1 gene(s), or
mutant thereof, can be introduced into one or more cells of the subject, a
somatic or germline
cell(s), in vivo or ex vivo (wherein cell(s) are removed from the subject, the
IF1 gene(s)
and/or mutant thereof is introduced into the cell(s), and the cell(s) are
returned back into the
subject afterwards), wherein optionally the introduced nucleotide sequence,
including the IF1
gene(s) (or mutant thereof) and/or regulatory sequence(s), is introduced as
naked nucleotide
material (optionally, without restriction, by one or more of heat shock, cell
squeezing, using
calcium phosphate to bind the DNA, electroporation, gene gun, sonoporation,
photoporation,
magnetofection, magnet assisted transfection, lipofection, impalefection,
optical transfection,
nucleofection, protofection, hydroporation, hydrodynamic delivery,
microinjection {DNA is
injected through the cell's nuclear envelope directly into the nucleus},
pronuclear injection
{after the sperm enters the egg, but before the genetic material of the sperm
and egg fuse,
genetic material is injected into the pronucleus of either the sperm or egg,
when these
pronuclei become visible, which is the first sign of successful fertilization;
this oocyte is then
.. implanted in the oviduct of a pseudopregnant female [e.g. induced when
female is bred by an
infertile male], the offspring thence carry the genetic modification},
embryonic Stem Cell-
Mediated Gene Transfer {gene is transfected into an embryonic stem cell(s)
that is then
inserted into a subject blastocyst(s), which is then implanted into foster
mother, resulting
offspring is/are chimeric, further mating can produce mice fully transgenic
subject(s) with the
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gene of interest}) or in a vector, wherein the vector can be, without
restriction, one or more of
a plasmid, episome, lipoplex (optionally with cationic lipids), liposome,
cationic liposome,
liposome coated in a polymer called polyethylene glycol, polymersome
(synthetic liposome,
made of amphiphilic block copolymers; copolymer is a polymer derived from more
than one
species of monomer), polyplex (complex of polymer with DNA), dendrimer (highly
branched
macromolecule with a spherical shape), inorganic nanoparticle (e.g. gold,
silica, iron oxide,
calcium phosphate etc.), organic nanoparticle, fugene, cell-penetrating
peptide, virosome
(viral envelope {phospholipid membrane and surface glycoproteins} of a virus
that, instead
of viral genetic material, contains therapeutic agent(s) of this invention
e.g. nucleotide
sequence(s) of the invention), virus (replication-competent, or replication-
defective/attenuated {coding region(s) for gene(s) necessary for virion
replication/packaging
is replaced with other gene(s), corrupted or deleted}), retrovirus (has a
single stranded RNA
genome that codes for a reverse transcriptase enzyme that produces a double
stranded DNA
sequence that is inserted into host genome by viral enzyme integrase; most
retroviruses can
only insert into actively dividing cells, which makes them appropriate for
delivering IF1
gene(s), or mutant thereof, for anti-cancer therapy; however, they tend to
insert into the
genome unpredictably, so "insertional mutagenesis" is a concern, however, in
an
embodiment, sequence(s) is incorporated that directs a retroviral coded DNA
sequence(s) to
be inserted at a specific locus or loci in the genome e.g. utilizing the beta-
globin locus control
region and/or by inserting a sequence for a Zinc finger nuclease which cuts
the genome at a
place that the viral DNA is desired to be inserted; retroviral vector
particles with tropism for
various target cells have been designed in the art), lentivirus (a genus of
retrovirus; can insert
into non-dividing cells; HIV is an example; enhancers can be used to improve
transduction
efficiency such as, without limitation, polybrene, protamine sulfate,
retronectin, and DEAE
Dextran), gammaretrovirus (a genus of retrovirus; non-limiting e.g. moloney
retrovirus),
adenovirus (doesn't integrate into genome, not replicated during cell
division), adeno-
associated virus (AAV; single stranded DNA virus {becomes double stranded in
the cell};
can be integrated into genome, mostly at a known site in humans {designated
AAVS1, in
human chromosome 19}, but is more likely to remain episomal {replicating
without
incorporation into the chromosome}; useful for dividing and non-dividing
cells; has been
used in clinical trials e.g. for retinal gene therapy by subretinal and/or
intravitreal injection
{e.g. Voretigene neparvovec (Luxturna)}, wherein an invention embodiment is to
deliver an
IF I gene(s), or mutant thereof, similarly to treat/ameliorate/prevent/combat
an aging-
related/linked/correlated eye(s) disease/condition, optionally age-related
macular
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degeneration, AMD; all AAV serotypes are [contemplated by/ componentry to] the
present
invention), self-complementary adeno-associated virus (scAAV; viral vector
engineered from
the naturally occurring adeno-associated virus (AAV); double stranded DNA
virus),
pseudotyped viruses (in which the endogenous viral envelope proteins have been
replaced by
either envelope proteins from other viruses, or by chimeric proteins, e.g. to
alter the cell
type(s) that the virus infects), hybrid vector (=genetically engineered virus,
to have desired
vector characteristics), recombinant virus, replication-deficient Herpes
simplex virus (human
neurotropic virus, infects neurons), cell penetrating peptide (e.g. protein
transduction domain
of Tat protein of HIV-1 virus) etc. This list is not exhaustive, other methods
of introducing
new genetic material to a subject are known to those of the art. Including
using an ancestral
or ancestral like AAV, such as Anc80 [368], which has significant merit
because it doesn't
stimulate immune reactions, because modern organisms haven't encountered this
virus for
generations.
Componentry to this invention is to use genome editing with engineered
nuclease(s) and/or
CRISPR/Cas9 and/or CRISPR/Cpfl and/or Zinc finger nuclease(s) and/or Zinc-
finger
nickase(s) and/or Transcription activator-like effector nuclease(s) (TALEN)
and/or
meganuclease(s) and/or megaTAL(s) and/or homing endonuclease(s) and/or
restriction
enzyme(s) and/or endonuclease and/or nuclease(s) and/or "gene targeting" (in
particular
"gene knock-in", replacement strategy based on homologous recombination)
and/or
Recombinant AAV mediated genome engineering (rAAV) and/or Multiplex Automated
Genomic Engineering (MAGE) and/or Cre-Lox system(s) and/or Flp-FRT system(s)
and/or
similar and/or other genetic editing/engineering technology to edit a native
IF I gene in a
subject to become a different IF I gene of this invention, optionally with a
nucleotide
change(s) that causes an amino acid residue change(s) at the "phosphorylation
control
switch" (optionally serine to alanine substitution) and/or the "pH dependence
motif'
(optionally a H49K substitution at position 49, using mature {mitochondrial
import signal
sequence cleaved off) human IF1 protein numbering) in the IF1 protein coded
for (these IF I
elements are defined in Figure 29), and/or to insert a nucleotide sequence(s)
of this
invention, which is a nucleotide sequence(s) that codes for one or more of the
IF1 protein
sequence embodiments of this invention, into a genome, optionally in one or
more cells of a
subject, optionally one or more of a somatic cell(s), germline cell(s),
gamete(s) (e.g. sperm(s)
or egg(s)), gametocyte(s), spermatocyte(s), oocyte(s), fertilized egg(s),
pronuclear fertilized
egg(s), embryonic stem cell(s), induced pluripotent stem cell (IPSC), or
other(s).
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Componentry to this invention is to "knock-in", wherein this term is well
understood in the
art (e.g. one of the art is very familiar with "knock-out" and "knock in"
mice, and protocols
used), an IF1 protein coding nucleotide DNA sequence of this invention,
optionally with a
nucleotide change(s) that causes an amino acid residue change(s) at the
"phosphorylation
control switch" (optionally serine to alanine substitution) and/or the "pH
dependence motif"
(optionally a H49K substitution at position 49, using mature {mitochondrial
import signal
sequence cleaved off} human IF1 protein numbering) in the IF1 protein coded
for (these IF1
elements are defined in Figure 29), in place of the native IF1 gene of the
subject.
Manipulating IF1 gene expression
An invention embodiment is to manipulate/change the
promoter/enhancer/repressor/regulatory sequence(s) of an IF1 gene(s), to
increase the
expression of IF1 protein, in somatic and/or germline cell(s) of a subject,
optionally wherein
a new promoter/enhancer/regulatory sequence(s) is added, optionally wherein
this increased
IF1 expression reduces FiFo ATP hydrolysis and slows the metabolic/aging rate
in this cell(s)
of the subject, optionally wherein exogenous/ambient heat is administered to
the subject to
substitute for this lesser heat generation and/or greater body insulation
(e.g. clothing) is
administered to the subject to reduce heat loss per unit time, which
correspondingly keeps the
subject warm despite lesser heat generation.
Summary/overview/recap of IF1 genetic manipulations of the invention
An embodiment of the invention is to change the sequence of a subject's IF1
gene, optionally
conferring a greater inhibitory potency against FIR) ATP hydrolysis at normal
mitochondrial
matrix pH 8, and/or increase the amount of IF1 protein in a cell(s) of a
subject, optionally in
all or the majority of cells in the subject or a subset of cell type(s) and/or
cell(s) in certain
tissue(s)/area(s) of the subject, optionally wherein change in the amount of
IF1 protein is
effected by (1) altering the regulatory sequence(s) of the subject's IF1 gene
to effect greater
IF1 gene expression, by greater transcription and/or translation, and/or (2)
by adding another
IF1 gene(s) to the subject's cell(s) genome, optionally wherein one or more of
the added IF!
gene(s) are from a different (larger and/or longer living {greater maximal
lifespan}) species
and/or is a mutant IF1 gene, optionally with a greater inhibitory potency
against FiFo ATP
hydrolysis at normal mitochondrial matrix pH 8, optionally with a nucleotide
change(s) that
causes an amino acid residue change(s) at the "phosphorylation control switch"
(optionally
serine to alanine substitution) and/or the "pH dependence motif' (optionally a
H49K
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substitution at position 49, using mature {mitochondria] import signal
sequence cleaved off}
human IF1 protein numbering) in the IF1 protein coded for (these IF1 elements
are defined in
Figure 29), optionally, especially if many/all of the subject's cells have
been manipulated in
this way, greater exogenous/ambient heat is administered to the subject to
substitute for lesser
endogenous heat generation and/or greater body insulation (e.g. clothing) is
administered to
the subject to reduce heat loss per unit time, which correspondingly keeps the
subject warm
despite lesser metabolic rate/body heat generation.
Example embodiment: IF1 gene therapy for Parkinson's disease
Firstly, gene therapy is now well established in the clinic, with thousands of
clinical trials
having taken place, with more being conducted, across dozens of countries for
many
indications, with notable approvals [369]. Secondly, for closer context, there
are numerous
reviews of gene therapy for Parkinson's disease in the literature e.g. [370,
371, 372, 373]. An
invention embodiment is to locally deliver, optionally bilaterally, optionally
by injection into
the subject's putamen (as used in [374]) and/or into the subject's basal
ganglia/substantia
nigra (SN)/substantia nigra pars compacta (SNpc), an IF1 gene(s) {optionally
another gene(s)
also, optionally one or more genes coding for an enzyme in dopamine synthesis
e.g. for
tyrosine hydroxylase, this enzyme catalyses the rate-limiting step of dopamine
synthesis, e.g.
for aromatic L-amino acid decarboxylase (AADC)}, optionally in a viral vector,
optionally in
a lentiviral viral vector (optionally ProSavin [374]) and/or an adeno-
associated viral vector
(optionally AAV2 [375]), to dopaminergic neurons, which reduces their FiFo ATP
hydrolysis, especially if the resulting extra IF I protein has an amino acid
residue that can't be
phosphorylated (optionally alanine) at its "phosphorylation control switch"
position and/or a
H49K substitution (position 49 using mature {mitochondrial import signal
sequence cleaved
off} human IF1 protein numbering) in its "pH dependence motif', which slows
the
metabolic/aging rate in dopaminergic neurons, which reduces the subject's
risk/progression
of Parkinson's disease, wherein these dopaminergic neurons generate less
metabolic heat, but
this is substituted for by heat transfer from surrounding body areas, such
that the
dopaminergic neurons are maintained at normal body temperature, optionally
wherein this
therapy is administered alongside one or more other therapies for Parkinson's
disease known
to those of the art e.g. L-DOPA administration, deep brain stimulation etc. In
other
embodiments, additionally or instead of, an IF I gene(s) is administered to
another brain
region(s), optionally a brain region(s) (whose dysfunction is) implicated in
Parkinson's
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CA 3050553 2019-07-25
disease e.g. optionally by infusion directly into the subject's subthalamic
nucleus (as used in
[374]).
Example embodiment: IF1 gene therapy for Osteoarthritis
Example embodiment: IF1 gene therapy extends mouse lifespan
An IF1 gene(s), optionally with a nucleotide change(s) that causes an amino
acid residue
change(s) at the "phosphorylation control switch" and/or the "pH dependence
motif' in the
IF1 protein coded for (these IF1 protein elements are defined in Figure 29),
with a promoter
that can drive its expression, optionally the CMV enhancer element and a
chicken 13-actin
promoter containing its first intron (regulatory element used in [376];
alternatively IF1 gene
is flanked by -actin promoter and terminator), is delivered to a nascent
B6(B6C3F1) mouse
embryo by pronuclear microinjection, wherein the resulting mouse has this
extra IF1 gene(s)
(with regulatory element(s) for expression) in all its cells (checked by PCR
for a number of
different cells from different tissues isolated from the mouse, amount of IF1
protein is also
assayed, wherein the genetically modified mouse should have more IF1 protein
in its cells).
This protocol is repeated to generate a number of such genetically modified
mice and their
lifespans, and/or those of their progeny as they are mated with each other
and/or backcrossed
with C57BL/6J mice one or more times, are monitored as compared to control
mice (no
genetic modification), wherein the genetically modified mice live longer:
longer health- and
life- (median and maximal) span. Food intake and body weight is monitored for
all mice over
their lifespan. Mice are kept in specific pathogen free conditions. Optionally
the study is
repeated or run in parallel with mice of a different genetic background. More
details on the
protocol to follow (e.g. number of mice and statistics to be used, healthspan
assays that can
be used {e.g. cardiac pathology, cataract development, oxidative damage,
mitochondrial
deletions etc.) etc.) can be sourced from emulating the protocol of [376]. A
protocol of a
different study of genetic manipulation and lifespan in the literature can be
used instead e.g.
[377]. But crucially, distinctly, all mice in the present study are housed at
37 C. The
genetically modified mice have more IF1 protein in their cells, less FiFo ATP
hydrolysis per
unit time, less oxidative phosphorylation per unit time, less heat generation
per unit time (not
detrimental when ambient temperature is 37 C), less ROS production per unit
time, less
oxidative damage per unit time, slower aging and longer lifespan.
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Instead of the 'IF1 transgene(s) adding an extra IF1 gene copy or copies to
the mouse genome,
as above, in an alternative invention embodiment, the IF1 transgene replaces
the native IF1
gene by a "genetic knock-in" (replacement strategy based on homologous
recombination),
optionally wherein the IF1 transgene has a nucleotide change(s) that causes an
amino acid
residue change(s) at the "phosphorylation control switch" and/or the "pH
dependence motif'
in the IF1 protein coded for (these IF I protein elements are defined in
Figure 29). These
mice also have slower aging and longer lifespan.
Example embodiment: IF1 gene therapy, restricted to brain, extends "brainspan"
Transgenic mice are made via pronuclear microinjection of mouse embryos with
an IF I
transgene(s), optionally with a nucleotide change(s) that causes an amino acid
residue
change(s) at the "phosphorylation control switch" and/or the "pH dependence
motif' in the
IF1 protein coded for (these IF1 protein elements are defined in Figure 29),
under the control
of human neurofilament heavy polypeptide (NEFH) promoter, or other
brain/neuron/astrocyte
specific promoter of the art, some (non-limiting) examples of which have been
mentioned
herein, optionally using multiple IF I gene copies with different promoters to
ensure IF1
transgene expression in both neurons and astrocytes. The mice are housed at an
ambient
temperature between 21 and 37 C. These genetically modified mice have more IF1
protein in
their brain, wherein there is less FIFO ATP hydrolysis per unit time, less
oxidative
phosphorylation per unit time, less heat generation per unit time (not
detrimental when
ambient temperature is 37 C, and not detrimental when ambient temperature is
lower because
heat transfer from the rest of the body, especially via blood flow, maintains
the brain at
37 C), less ROS production per unit time, less oxidative damage per unit time,
slower brain
aging and longer "brainspan" i.e. less cognitive decline with age, lower
risk/later
onset/slower progression of age-related/correlated brain disease(s) e.g.
Alzheimer's disease,
dementia etc.
CONSOLIDATORY FORMULA/MECHANISM
This Formula consolidates prior formulas to present some preferred embodiments
of the
invention. These structures share the common motif:
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CA 3050553 2019-07-25
(X3)
NsN
(x2xx)xx
X1
especially.
N' NH
N7:21 Or
NNNN.N
X1 Xi
Wherein each X3 is, independently at each point of use, absent, or hydrogen,
or alkyl, or
substituted alkyl (non-limiting example: CF3), or deuterated alkyl (non-
limiting example:
CD3), or aminoalkyl, or thioalkyl, or alkoxy, or haloalkyl, or haloalkoxy, or
hydroxyalkyl, or
any atom or isotope permitted by valence (including any accompanying hydrogens
by
valence e.g. (non-limiting) OH, NH2, SH, SiH3, PH2 etc.), for example
deuterium, or halogen,
or fluorine;
Each X2" is, independently at each point of use, selected from a single bond,
0, S. Se, NXP,
PXP, BXP, C(XP)2 or Si(XP)2, wherein each XP is, independently at each point
of use, selected
from a constituent group of X3 (defined earlier);
xx and xy are each independently selected from 0, 1, 2, 3, 4, 5;
X 1 is
R2N/R3
/INN.
N
(R4)q R1
wherein Markush symbols are as defined previously for Formula (I), or
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CA 3050553 2019-07-25
(R1),õ
(R7)q
\ Aa,
N¨Z
1-- N \
R6
L
Bb
\
(R5)õ
wherein Markush symbols are as defined previously for Formula (II), or
R12
I
(Xk)k
R1 R2
L
L
R3
Y
R5 R4
wherein Markush symbols are as defined previously for Formula (III), or
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CA 3050553 2019-07-25
R2 R1 X NOM*
R3 fi (R1),,
I 8
G1
R6 R7
R4 R5 ( n
)m
Z
wherein Markush symbols are as defined previously for Formula (IV), or
0 ......Ar 0
AAA ¨Nly\ /yNH AAA
N
H
MR 0 MR
wherein each AAR is, independently at each point of use, an amino acid side
chain,
optionally (but not restrictively) the side chain of an amino acid coded for
by the genetic
code, each AAA is, independently at each point of use, an amino acid
(proteingenic or non-
proteingenic) or a chain of amino acids (proteingenic, or non-proteingenic or
a combination)
linked by peptide bonds, not exceeding 300 amino acid residues, optionally an
amino acid
sequence coded for by a component of a genome, optionally an amino acid chain
sub-
sequence of an IF I protein sequence, preferably an amino acid sequence
falling under
Formula (VII) and/or presented in Figure 29, optionally wherein one or more of
the amino
acids have a post-translational modification(s), and/or a
modification/manipulation to
increase plasma stability, wherein such strategies are well known to those of
the art e.g.
swopping stereochemistry (D instead of L) of one or more amino acids,
optionally with a
modification at their N and/or C terminal ends as disclosed elsewhere herein.
Whilst it is true that compounds of Formula (V) don't have this imidazole
structural motif, it
is also true that they aren't particularly potent inhibitors of FIR ATP
hydrolysis.
IF I monomers and [Fl dimers can inhibit FIR ATP hydrolysis because their FiFo
ATP
hydrolysis domain is exposed. Higher IF I oligmers (>dimer) cannot inhibit
FIR) ATP
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CA 3050553 2019-07-25
hydrolysis because their FIFO ATP hydrolysis domain is buried in the IF1
oligomer, thence
higher oligomerization (>dimer) sequesters IF I from inhibiting FiFo ATP
hydrolysis, and this
is the basis to the pH dependence of IF1 inhibition of F1 F0 ATP hydrolysis:
higher pH = IF1
higher (>dimer) oligomerization and low FIR ATP hydrolysis inhibition, lower
pH = higher
(>dimer) IF1 oligomers break up and high FIR ATP hydrolysis inhibition.
Without wishing
to be restricted by theory, how this imidazole motif enables a compound to
inhibit FiFo ATP
hydrolysis is that it interacts with, and locates the compound to, one or more
histidines in the
pH dependence motif of IF I (Figure 29B) and its location here blocks the
higher
oligomerization (>dimer) of IF1, without blocking the FIFO ATP hydrolysis
domain of IFI,
and so there are more free IF1 monomers and dimers, thence more inhibition of
FiFo ATP
hydrolysis. So, the compound interacts with, and exerts its action through, IF
I rather than
ATP synthase. The compound can reduce FIFO ATP synthesis, but this is by
uncoupling
(Figure 31) rather than by interaction with ATP synthase and direct inhibition
of FIFO ATP
synthesis. Componentry to this invention is any compound(s) that interacts
with IF1,
optionally the pH dependence motif of IF I (Figure 29B), optionally wherein
the
compound(s) has an imidazole motif, to reduce/prevent/stop the higher (>dimer)
oligomerization of IF1, optionally to treat/prevent/ ameliorate/combat one or
more of the
diseases or disorders or physiological processes or sub-optimalities mentioned
herein,
optionally cancer and/or aging.
With a compound(s) of this invention bound, IF I should still be able to
inhibit FiFo ATP
hydrolysis, but with the compound bound, IF I can't higher (>dimer)
oligomerize and so there
is more free IF] monomer/dimer and so greater inhibition of FiFo ATP
hydrolysis. Thence
the size/shape of the compound is paramount, such that is blocks/interferes
with the higher
(>dimer) oligomerization, and not the inhibitory domain, of IFI. The higher
(>dimer)
oligomerization domain (or part thereof, or modified form thereof) of IF!,
without the
inhibitory domain of IF I attached, is componentry to this invention (Figure
29).
A method to find a compound of this invention is to assess whether a compound
reduces/prevents/stops the higher (>dimer) oligomerization of IF1 at pH 8. If
so, in an
optional but preferable rd step, it is assessed whether the compound doesn't
(necessary)
block IF1 inhibition of FIFO ATP hydrolysis in a sub-mitochondrial particle
(SMP) assay,
optionally conducted at pH 6.7. In a 3rd step, or as an alternative rd step,
it is assessed
whether the compound enhances (preferable) IF I inhibition of FiFo ATP
hydrolysis in a sub-
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mitochondrial particle (SMP) assay, optionally conducted at pH 8, optionally
wherein action
through IF I, rather than direct action upon ATP synthase, can be assayed by
observing the
effect of removing IF I from the SMP assay. At pH 8, if there is much less FIR
ATP
hydrolysis inhibition with much less IF1 in the assay, then the compound is
acting upon FIFO
ATP hydrolysis via interaction with IF1 rather than directly upon ATP
synthase. If a
compound hits criterion/criteria in one or more of these steps, and most
preferably in all 3 of
these steps, then it is a compound of this invention. In some embodiments a
screening method
is employed using one or more of these steps, wherein a number of compounds
are tested in
this method to find one or more compounds that reduces/prevents/stops the
higher (>dimer)
oligomerization of IF I at pH 8 and increases IF I inhibition of F IF ATP
hydrolysis at pH 8.
The 1st step of this method is especially suited to high throughput screening,
and only those
compounds that pass the 1st step need be entered into the more
technically/time demanding
2' and/or 3' steps. In some embodiments one or more of these steps is perfomed
at the
European Lead Factory.
Definitions used to specify Formulas (I), (H), (III), (IV), (V), (VI) and
(VII)
The initial definition provided for a group or term herein applies to that
group or term
throughout the present specification, individually or as part of another
group, unless
otherwise indicated.
The term "alkyl" refers to straight or branched chain hydrocarbon groups
having 1 to 21
carbon atoms, preferably 1 to 8 carbon atoms. Lower alkyl groups, that is,
alkyl groups of 1
to 4 carbon atoms, are most preferred.
The term "substituted alkyl" refers to an alkyl group as defined above having
one, two, three,
or four substituents selected from the group consisting of PH2, deuterium,
halogen,
trifluoromethyl, alkenyl, alkynyl, nitro, cyano, CH, keto (=0), ORa, SRa,
NRaRb, NRaS02,
NRaSO2Rc, SO2Rc, SO2NRaRb, CO2Ra, C(=0)Ra, C(=0)NRaRb, OC(=0)Ra, ¨
OC(=0)NRaRb, NRaC(=0)Rb, NRaCO2Rb, =N¨OH, =N-0-alkyl, aryl, heteroaryl,
heterocyclo and cycloalkyl, wherein Ra and Rb are selected from hydrogen,
alkyl, alkenyl,
cycloalkyl, heterocyclo, aryl, and heteroaryl, and Rc is selected from
hydrogen, alkyl,
cycloalkyl, heterocyclo aryl and heteroaryl. When a substituted alkyl includes
an aryl,
heterocyclo, heteroaryl, or cycloalkyl substituent, said ringed systems are as
defined below
and thus may in turn have zero to four substituents (preferably 0-2
substituents), also as
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CA 3050553 2019-07-25
defined below. When either Ra, Rb or fic is an alkyl, said alkyl may
optionally be substituted
with 1-2 of PH2, deuterium, halogen, trifluoromethyl, alkenyl, alkynyl, nitro,
cyano, CH, keto
(=0), OH, 0(alkyl), phenyloxy, benzyloxy, SH, S(alkyl), NH2, NH(alkyl),
N(alkyl)2, NHS02,
NHS02(alkyl), S02(alkyl), SO2NH2, SO2NH(alkyl), CO2H, CO2(alkyl), C(0)H,
C(0)alkyl, C(=0)NH2, C(=0)NH(alkyl), C(=0)N(alky1)2, OC(=0)alkyl, ¨0C(=0)NH2,¨
OC(=0)NH(alkyl), NHC(=0)alkyl, and/or NHCO2(alkyl).
"Alkyl" when used in conjunction with another group such as in arylalkyl
refers to a
substituted alkyl in which at least one of the substituents is the
specifically named group. For
example, the term arylalkyl includes benzyl, or any other straight or branched
chain alkyl
having at least one aryl group attached at any point of the alkyl chain. As a
further example,
the term carbamylalkyl includes the group ¨(CH2)n¨NH¨C(=0)alkyl, Wherein n is
1 to
12.
The term "alkenyl" refers to straight or branched chain hydrocarbon groups
having 2 to 21
carbon atoms and at least one double bond. Alkenyl groups of 2 to 6 carbon
atoms and having
one double bond are most preferred.
The term "alkynyl" refers to straight or branched chain hydrocarbon groups
having 2 to 21
carbon atoms and at least one triple bond. Alkynyl groups of 2 to 6 carbon
atoms and having
one triple bond are most preferred.
The term "alkylene" refers to bivalent straight or branched chain hydrocarbon
groups having
1 to 21 carbon atoms, preferably 1 to 8 carbon atoms, e.g., {¨CH2¨}n, Wherein
n is 1 to 12,
preferably 1-8. Lower alkylene groups, that is, alkylene groups of 1 to 4
carbon atoms, are
most preferred. The terms "alkenylene" and "alkynylene" refer to bivalent
radicals of alkenyl
and alknyl groups, respectively, as defined above.
When reference is made to a substituted alkylene, alkenylene, or alkynylene
group, these
groups are substituted with one to four substituents as defined above for
alkyl groups. A
substituted alkylene, alkenylene, or alkynylene may have a ringed substituent
attached in a
spiro fashion as in
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`,..,...%5L 7 %.**=,..57=....s.7....
f
and so forth.
The term "alkoxy" refers to an alkyl or substituted alkyl group as defined
above having one,
two or three oxygen atoms (-0¨) in the alkyl chain. For example, the term
"alkoxy"
includes the groups ¨0¨CI-12alkyl, ¨C1_6alkylene-O¨C1.6alkyl, ¨C1_4alkylene-0-
phenyl,
and so forth.
The term "thioalkyl" or "alkylthio" refers to an alkyl or substituted alkyl
group as defined
above having one or more sulphur (¨S¨) atoms in the alkyl chain. For example,
the term
"thioalkyl" or "alkylthio" includes the groups ¨(CH2), ¨S¨CH2aryl,
¨(CH2)n¨S¨aryl,
etc. etc.
The term "aminoalkyl" or "alkylamino" refers to an alkyl or substituted alkyl
group as
.. defined above having one or more nitrogen (¨N11.`¨) atoms in the alkyl
chain. For example,
the term "aminoalkyl" includes the groups ¨NR`¨C1..12alkyl and ¨CH2¨NR`-aryl,
etc.
(where R' is hydrogen, alkyl or substituted alkyl as defined above.) "Amino"
refers to the
group ¨N H2.
When a subscript is used as in Ci_salkyl, the subscript refers to the number
of carbon atoms
the group may contain. Zero when used in a subscript denotes a bond, e.g., CO-
4 alkyl refers to
a bond or an alkyl of 1 to 4 carbon atoms. When used with alkoxy, thioalkyl or
aminoalkyl, a
subscript refers to the number of carbon atoms that the group may contain in
addition to
heteroatoms. Thus, for example, monovalent. C1_2aminoalkyl includes the groups
¨CH2-
__________________________________________________ NH2, ¨NH¨C H3, ¨(C H2)2¨N
H2, ¨NH¨C H2¨C H3, ¨CH2¨NH2 CH3, and ¨N¨
(CH3)2. A lower aminoalkyl comprises an aminoalkyl having one to four carbon
atoms.
The alkoxy, thioalkyl, or aminoalkyl groups may be monovalent or bivalent. By
"monovalent" it is meant that the group has a valency (i.e., power to combine
with another
group), of one, and by "bivalent" it is meant that the group has a valency of
two. For
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example, a monovalent alkoxy includes groups such as ¨0¨C1-12alkyl,
¨Ci_6alkylene¨
C1.6alkyl, etc., whereas a bivalent alkoxy includes groups such as
¨0¨C1_2alkylene-, ¨
C _6alky lene-O¨C 1_6alkylene-, etc.
The term "acyl" refers to a carbonyl
(ii)
linked to an organic group i.e.
0
I I
Rd
wherein Rd may be selected from alkyl, alkenyl, substituted alkyl, substituted
alkenyl, aryl,
.. heterocyclo, cycloalkyl, or heteroaryl, as defined herein.
The term "alkoxycarbonyl" refers to a group having a carboxy or ester group
ii
0¨
linked to an organic radical, i.e.,
0
I I 0
Rd
Wherein Rd is as defined above for acyl.
The term "carbamyl" refers to a functional group in which a nitrogen atom is
directly bonded
to a carbonyl, i.e., as in ¨NReC(=0)Rf or ¨C(=0)NReRe, wherein Re and Re can
be
hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy,
cycloalkyl, aryl,
heterocyclo, or heteroaryl, or they may join to form a ring.
The term "sulfonyl" refers to a sulphoxide group (i.e.,¨S(0)1_2) linked to an
organic radical
Re, as defined above.
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The term "sulfonamide" or "sulfonamido" refers to the group -S(0)2NReRf,
wherein Re and
Rf are as defined above. Preferably when one of Re and Rf is optionally
substituted heteroaryl
or heterocycle (as defined below), the other of Re and Rf is hydrogen or
alkyl.
The term "cycloalkyl" refers to fully saturated and partially unsaturated
hydrocarbon rings of
3 to 9, preferably 3 to 7 carbon atoms. The term "cycloalkyl" includes such
rings having zero
to four substituents (preferably 0-2 substituents), selected from the group
consisting of OH,
SH, PH2, deuterium, halogen, alkyl, substituted alkyl (e.g., trifluoromethyl),
alkenyl,
substituted alkenyl, alkynyl, nitro, cyano, CH, keto, ORd, SRd NRafte NRcS02,
NReS02Re,
C(=0)H, acyl, -CO2H, alkoxycarbonyl, carbamyl, sulfonyl, sulfonamide, -
0C(=0)Rd,
=N-OH, =N-0-alkyl, aryl, heteroaryl, heterocyclo, a 4 to 7 membered
carbocyclic ring,
and a five or six membered ketal, e.g., 1,3-dioxolane or 1,3-dioxane, wherein
Re, Rd and Re
are defined as above. The term "cycloalkyl" also includes such rings having a
phenyl ring
fused thereto or having a carbon-carbon bridge of 3 to 4 carbon atoms.
Additionally, when a
cycloalkyl is substituted with a further ring, i.e., aryl, arylalkyl,
heteroaryl, heteroarylalkyl,
heterocyclo, heterocycloalkyl, cycloalkylalkyl, or a further cycloalkyl ring,
such ring in turn
may be substituted with one to two of Co_aalkyl optionally substituted with
OH, SH, PH2,
halogen, trifluoromethyl, alkenyl, alkynyl, nitro, cyano, CH, keto (=0), OH,
0(alkyl),
phenyloxy, benzyloxy, SH, S(alkyl), NH2, NH(alkyl), N(alkyl)2, NHS02,
NHS02(alkyl),
S02(alkyl), SO2NH2, SO2NH(alkyl), CO2H, CO2(alkyl), C(=0)H, C(=0)alkyl,
C(=0)NH2,
C(=0)NH(alkyl), C(0)N(alkyl)2, OC(=0)alkyl, -0C(=0)NH2, -0C(=0)NH(alkyl),
NHC(=0)alkyl, and NHCO2(alkyl).
The term "halo" or "halogen" refers to chloro, bromo, fluoro and iodo.
The term "haloalkyl" means a substituted alkyl having one or more halo
substituents. For
example, `taloallcyl" includes mono, bi, and trifluoromethyl.
The term "haloallcoxy" means an alkoxy group having one or more halo
substituents. For
example, "haloalkoxy" includes OCF3.
The term "aryl" refers to phenyl, biphenyl, I-naphthyl, 2-naphthyl, and
anthracenyl, with
phenyl being preferred. The term "aryl" includes such rings having zero to
four substituents
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(preferably 0-2 substituents), selected from the group consisting of
deuterium, OH, SH,
PH2, halo, alkyl, substituted alkyl (e.g., trifluoromethyl), alkenyl,
substituted alkenyl, alkynyl,
nitro, cyano, CH, ORd, SRd, NRdRe, NRdS02, NRdS02Re, C(0)H, acyl, -CO2H,
alkoxycarbonyl, carbamyl, sulfonyl, sulfonamide, -0C(=0)Rd, heteroaryl,
heterocyclo,
cycloalkyl, phenyl, benzyl, napthyl, including phenylethyl, phenyloxy, and
phenylthio,
wherein Re, Rd and Re are defined as above. Additionally, two substituents
attached to an
aryl, particularly a phenyl group, may join to form a further ring such as a
fused or spiro-ring,
e.g., cyclopentyl or cyclohexyl or fused heterocycle or heteroaryl. When an
aryl is substituted
with a further ring, such ring in turn may be substituted with one to two of
Co-aalkyl
optionally substituted with deuterium, SH, PH2, halogen, trifluoromethyl,
alkenyl, alkynyl,
nitro, cyano, CH, keto (=0), OH, 0(alkyl), phenyloxy, benzyloxy, SH, S(alkyl),
NH2,
NH(alkyl), N(alkyl)2, NHS02, NHS02(alkyl), S02(alkyl), SO2NH2, SO2NH(alkyl),
CO2H,
CO2(alkyl), C(=0)H, C(=0)alkyl, C(=0)NH2, C(=0)NH(alkyl), C(=0)N(alky1)2,
OC(=0)alkyl, -0C(=0)NH2, -0C(=0)NH(alkyl), NHC(=0)alkyl, and NHCO2(alkyl).
The term "heterocyclo" refers to substituted and unsubstituted non-aromatic 3
to 7 membered
monocyclic groups, 7 to 11 membered bicyclic groups, and 10 to 15 membered
tricyclic
groups, in which at least one of the rings has at least one heteroatom
selected from 0, S and
N. Each ring of the heterocyclo group containing a heteroatom can contain one
or two oxygen
or sulfur atoms and/or from one to four nitrogen atoms provided that the total
number of
heteroatoms in each ring is four or less, and further provided that the ring
contains at least
one carbon atom. The fused rings completing bicyclic and tricyclic groups may
contain only
carbon atoms and may be saturated, partially saturated, or unsaturated. The
nitrogen and
sulfur atoms may optionally be oxidized and the nitrogen atoms may optionally
be
quaternized. The heterocyclo group may be attached at any available nitrogen
or carbon
atom. The heterocyclo ring may contain zero to four substituents (preferably 0-
2
substituents), selected from the group consisting of deuterium, OH, SH, PH2,
halo, alkyl,
substituted alkyl (e.g., trifluoromethyl), alkenyl, substituted alkenyl,
alkynyl, nitro, cyano,
CH, keto, ORd, SRd, NRdRe, NRdS02, NReiSO2Rc, SO2Rd, C(0)H, acyl, -CO2H,
alkoxycarbonyl, carbamyl, sulfonyl, sulfonamide, -0C(0)Rd, =N-OH, =N-0-alkyl,
aryl, heteroaryl, cycloalkyl, a five or six membered ketal, e.g., 1,3-
dioxolane or 1,3-dioxane,
or a monocyclic 4 to 7 membered non aromatic ring having one to four
heteroatoms, wherein
Re, Rd and Re are defined as above. The term "heterocyclo" also includes such
rings having a
phenyl ring fused thereto or having a carbon-carbon bridge of 3 to 4 carbon
atoms.
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Additionally, when a heterocyclo is substituted with a further ring, i.e.,
aryl, arylalkyl,
heteroaryl, heteroarylalkyl,cycloalkyl, cycloalkylalkyl, heterocycloalkyl, or
a further
heterocyclo ring, such ring in turn may be substituted with one to two of
Co_aalkyl optionally
substituted with deuterium, SH, PH2, halogen, trifluoromethyl, alkenyl,
alkynyl, nitro, cyano,
CH, keto (=0), OH, 0(alkyl), phenyloxy, benzyloxy, SH, S(alkyl), NH2,
NH(alkyl),
N(alkyl)2, NHS02, NHS02(alkyl), S02(alkyl), SO2NH2, SO2NH(alkyl), CO2H,
CO2(alkyl),
C(=0)H, C(0)alkyl, C(=0)NH2, C(=0)NH(alkyl), C(=0)N(alky1)2, OC(=0)alkyl, -
OC(=0)NH2, -0C(=0)NH(alkyl), NHC(=0)alkyl, and NHCO2(alkyl).
Exemplary monocyclic groups include azetidinyl, pyrrolidinyl, oxetanyl,
imidazolinyl,
oxazolidinyl, isoxazolinyl, thiazolidinyl, isothiazolidinyl,
tetrahydrofuranyl, piperidinyl,
piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-
oxoazepinyl, azepinyl,
4-piperidonyl, tetrahydropyranyl, morpholinyl, thiamorpholinyl,
thiamorpholinyl sulfoxide,
thiamorpholinyl sulfone, 1,3-dioxolane and tetrahydro-1,1-dioxothienyl and the
like.
Exemplary bicyclic heterocyclo groups include quinuclidinyl.
The term "heteroaryl" refers to substituted and unsubstituted aromatic 5 to 7
membered
monocyclic groups, 9 or 10 membered bicyclic groups, and 11 to 14 membered
tricyclic
groups which have at least one heteroatom selected from 0, S and N in at least
one of the
rings. Each ring of the heteroaryl group containing a heteroatom can contain
one or two
oxygen or sulfur atoms and/or from one to four nitrogen atoms provided that
the total number
of heteroatoms in each ring is four or less and each ring has at least one
carbon atom. The
fused rings completing the bicyclic and tricyclic groups may contain only
carbon atoms and
may be saturated, partially saturated, or unsaturated. The nitrogen and sulfur
atoms may
optionally be oxidized and the nitrogen atoms may optionally be quaternized.
Heteroaryl
groups which are bicyclic or tricyclic must include at least one fully
aromatic ring but the
other fused ring or rings may be aromatic or non-aromatic. The heteroaryl
group may be
attached at any available nitrogen or carbon atom of any ring. The heteroaryl
ring system may
contain zero to four substituents (preferably 0-2 substituents), selected from
the group
consisting of deuterium, OH, SH, PH2, halo, alkyl, substituted alkyl (e.g.,
trifluoromethyl),
alkenyl, substituted alkenyl, alkynyl, nitro, cyano, CH, ORd, SRd, NRdRe,
NRdS02,
NRdS02Re, SO2Rd, C(=0)H, acyl, -CO2H, alkoxycarbonyl, carbamyl, sulfonyl,
sulfonamide, -0C(=0)Rd, heterocyclo, cycloalkyl, aryl, or a monocyclic 4 to 7
membered
aromatic ring having one to four heteroatoms, including phenylethyl,
phenyloxy, and
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phenylthio, wherein Itc, Rd and R, are defined as above. Additionally, when a
heteroaryl is
substituted with a further ring, i.e., aryl, arylalkyl, heterocyclo,
heterocycloalkyl, cycloalkyl,
cycloalkylalkyl, heteroarylalkyl, or a further heteroaryl ring, such ring in
turn may be
substituted with one to two of C0-4 alkyl optionally substituted with
deuterium, PH2, halogen,
trifluoromethyl, alkenyl, alkynyl, nitro, cyano, CH, keto (=0), OH, 0(alkyl),
phenyloxy,
benzyloxy, SH, S(alkyl), NH2, NH(alkyl), N(alkyl)2, NHS02, NHS02(alkyl)n,
S02(alkyl),
SO2NH2, SO2NH(alkyl), CO2H, CO2(alkyl), C(0)H, C(0)alkyl, C(=0)NH2,
C(=0)NH(alkyl), C(=0)N(alky1)2, OC(=0)alkyl,-0C(=0)NH2, ¨0C(=0)NH(alkyl),
NHC(=0)alkyl, and NHCO2(alkyl).
Exemplary monocyclic heteroaryl groups include pyrrolyl, pyrazolyl,
pyrazolinyl, imidazolyl,
oxazolyl, isoxazolyl, thiazolyl
(Le <;-),
thiadiazolyl, isothiazolyl, furanyl, thienyl, oxadiazolyl, pyridyl, pyrazinyl,
pyridinyl,
pyrimidinyl, pyridazinyl, triazinyl and the like.
Exemplary bicyclic heteroaryl groups include indolyl, benzothiazolyl,
benzodioxolyl,
benzoxaxolyl, benzothienyl, quinolinyl, tetrahydroisoquinolinyl,
isoquinolinyl,
benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, chromonyl,
coumarinyl,
benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl,
furopyridinyl,
dihydroisoindolyl, tetrahydroquinolinyl and the like.
Exemplary tricyclic heteroaryl groups include carbazolyl, benzidolyl,
phenanthrollinyl,
acridinyl, phenanthridinyl, xanthenyl and the like.
When the term "unsaturated" is used herein to refer to a ring or group, the
ring or group may
be fully unsaturated or partially unsaturated.
The phrase "optionally substituted" is intended to include substituted or
unsubstituted
possibilities. Accordingly, the phrase "each group of which may be optionally
substituted
means that each group includes both substituted and unsubstituted groups.
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The use of the phrase "Where valence allows" means that the groups may be
substituted only
to the degree and nature allowed by valency of the group. This is commonly
understood by
those of skill in the art. For example, a hydrogen substituent cannot be
further substituted nor
can a phenyl group be directly substituted by an oxo group due to limits on
valency.
The term "substituted amino" refers to a group of the formula -NZ2Z3 wherein
Z2 is
hydrogen, alkyl, cycloalkyl, aryl, arylalkyl, (cycloalkyl)alkyl,
morpholinylalkyl, heterocyclo
or (heterocyclo)alkyl and Z3 is hydrogen, alkyl, cycloalkyl, aryl, arylalkyl,
haloalkyl,
hydroxyalkyl, alkoxyalkyl, thioalkyl, (cycloalkyl)alkyl or hydroxyalkyl
further substituted
with a carboxylic ester or carboxylic acid, with the proviso that when Z2 is
hydrogen, then Z3
is other than hydrogen; or Z2 and Z3 taken together with the nitrogen atom to
which they are
attached are 1-pyrrolidinyl, 1-piperidinyl, 1-azepinyl, 4-morpholinyl, 4-
thiamorpholinyl, 1-
piperazinyl, 4-alkyl-1-piperazinyl, 4-arylalkyl-1-piperazinyl, 4-diarylalkyl-1-
piperazinyl; or
1-pyrrolidinyl, 1-piperidinyl, 1-azepinyl substituted with alkyl, alkoxy,
alkylthio, halo,
trifluoromethyl or hydroxy.
Hydroxy, hydroxyl and ¨OH are used interchangeably herein.
The term "heterocyclo" or "hetero" also includes such monocyclic and bicyclic
rings wherein
an available carbon atom is substituted with a (Cl-C4)-alkyl, aryl, (CI-C4)-
alkylthio, (Ci-C4)-
alkoxy, halo, nitro, keto, cyano, CH, hydroxy, azo, thiazo, amino, -NH-(Cl-C4)-
alkyl, -N((Ci-
C4)-alky1)2, -CF3, (aminoester)alkyl, carboxylic acid, carboxylic ester, -
OCHF2 or (Ci-C4)-
alkoxy further substituted with a carboxylic acid or such monocyclic and
bicyclic rings
wherein two or three available carbons have substituents selected from methyl,
methoxy,
methylthio, halo, -CF3, nitro, hydroxy, amino and -OCHF2.
Herein, when "carboxyl" or "carboxylic acid" is used, this can mean ¨C(0)0H
but can also
refer to carboxylic ester or ester, which encompasses -0C(0)0Rester and -
C(0)0R.
- -,ster
wherein Rester is selected from hydrogen, deuterium, alkyl, alkenyl, alkoxy,
thioalkyl,
aminoalkyl, cycloalkyl, haloalkyl, haloalkoxy, aryl, heteroaryl or
heterocyclo, each of which
may be optionally substituted, as defined herein.
Use of "aryl" herein encompasses "aryloxy", which refers to ¨0-aryl, wherein
aryl is selected
from prior definition of aryl specified herein. Use of "heteroaryl" herein
encompasses
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"heteroaryloxy", which refers to ¨0-heteroaryl, wherein heteroaryl is selected
from prior
definition of heteroaryl specified herein. Use of "heterocyclo" herein
encompasses
"hetereocyclooxy", which refers to ¨0-heterocyclo, wherein heterocyclo is
selected from
prior definition of heterocyclo specified herein.
Use of "ether" herein encompasses --0.,,R-ther wherein Rether is selected from
hydrogen,
deuterium, alkyl, alkenyl, alkoxy, thioalkyl, aminoalkyl, cycloalkyl,
haloalkyl, haloalkoxy,
aryl, heteroaryl or heterocyclo, each of which may be optionally substituted,
as defined
herein.
At various places herein, substituents of compounds are disclosed in groups or
in ranges. It is
specifically intended that the invention include each and every individual sub-
combination of
the members of such groups and ranges. For example, the term "Ci_3allcyl" is
intended to
include Cialkyl (methyl), C2alkyl (ethyl), C3alkyl.
As used herein, the terms "nucleic acid(s)" and "nucleotide sequence(s)" and
"polynucleotide(s)" include DNA molecules (e.g. cDNA or genomic DNA), RNA
molecules
(e.g. mRNA), combinations of DNA and RNA molecules or hybrid DNA/RNA
molecules,
and analogs, derivatives and modifications (e.g. methylation) of DNA or RNA
molecules.
Analogs can be generated using, for example, nucleotide analogs, which
include, but are not
limited to, inosine or tritylated bases. Such analogs can also comprise DNA or
RNA
molecules comprising modified backbones that lend beneficial attributes to the
molecules
such as, for example, nuclease resistance or an increased ability to cross
cellular membranes.
The nucleic acids or nucleotide sequences can be single-stranded (the sense or
antisense
strand), double-stranded, may contain both single-stranded and double-stranded
portions, and
may contain triple-stranded portions.
As used herein, the term "hybridizes under stringent conditions" describes
conditions for
hybridization and washing under which nucleotide sequences at least 30%
(preferably, at
least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least
60%, at least 65%, at
least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least
95%) identical to
each other typically remain hybridized to each other. Such stringent
conditions are known to
those skilled in the art and can be found in Current Protocols in Molecular
Biology, John
Wiley & Sons, N.Y. (1989). In one, non-limiting example stringent
hybridization conditions
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are hybridization at 6*Sodium chloride/sodium citrate (SSC) at about 45 C,
followed by one
or more washes in 0.1*SSC, 0.2% SDS at about 68 C. In a preferred, non-
limiting example
stringent hybridization conditions are hybridization in 6xSSC at about 45 C,
followed by one
or more washes in 0.2*SSC, 0.1% SDS at 50-65 C (i.e. one or more washes at 50
C, 55 C,
60 C. or 65 C). It is understood that the nucleic acids of the invention do
not include nucleic
acid molecules that hybridize under these conditions solely to a nucleotide
sequence
consisting of only A or T nucleotides.
As used herein, the term "proteinaceous agent" is a peptide, polypeptide,
protein, fusion
protein or antibody. Wherein "analog" refers to a proteinaceous agent that
possesses a
similar/identical function to a second proteinaceous agent, which does, or
does not, comprise
a similar/identical amino acid sequence. Two proteinaceous agents have a
similar amino acid
sequence, and/or 3D structure, if at least one of the following is satisfied:
(a) a proteinaceous
agent having an amino acid sequence that is at least 30%, at least 35%, at
least 40%, at least
45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at
least 75%, at least
80%, at least 85%, at least 90%, at least 95% or at least 99% identical to the
amino acid
sequence of a second proteinaceous agent; (b) a proteinaceous agent encoded by
a nucleotide
sequence that hybridizes under stringent conditions to a nucleotide sequence
encoding a
second proteinaceous agent of at least 5 contiguous amino acid residues, at
least 10
contiguous amino acid residues, at least 15 contiguous amino acid residues, at
least 20
contiguous amino acid residues, at least 25 contiguous amino acid residues, at
least 40
contiguous amino acid residues, at least 50 contiguous amino acid residues, at
least 60
contiguous amino residues, at least 70 contiguous amino acid residues, at
least 80 contiguous
amino acid residues, at least 90 contiguous amino acid residues, at least 100
contiguous
amino acid residues, at least 125 contiguous amino acid residues, or at least
150 contiguous
amino acid residues; (c) a proteinaceous agent encoded by a nucleotide
sequence that is at
least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least
55%, at least 60%, at
least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least
90%, at least 95% or
at least 99% identical to the nucleotide sequence encoding a second
proteinaceous agent; (d)
a proteinaceous agent with similar structure to a second proteinaceous agent
refers to a
proteinaceous agent that has a similar secondary, tertiary or quaternary
structure to the second
proteinaceous agent. The structure of a proteinaceous agent can be determined
by one or
more methods known to those skilled in the art, including but not limited to x-
ray
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crystallography, nuclear magnetic resonance (NMR) spectroscopy, circular
dichroism,
crystallographic electron microscopy, cryogenic electron microscopy (cryo-EM).
To determine the percent identity of two amino acid sequences, or of two
nucleic acid
sequences, the sequences are aligned for optimal comparison purposes (e.g.,
gaps can be
introduced in the sequence of a first amino acid, or nucleic acid, sequence
for optimal
alignment with a second amino acid, or nucleic acid, sequence). The amino
acid/nucleotide
residues at corresponding amino acid/nucleotide positions are then compared.
When a
position in the first sequence is occupied by the same amino acid residue or
nucleotide as the
corresponding position in the second sequence, then the molecules are
identical at that
position. The percent identity between the two sequences is a function of the
number of
identical positions shared by the sequences {i.e. % identity = (number of
identical
overlapping positions/total number of positions)*100}. The determination of
percent identity
between two sequences can also be accomplished using a mathematical algorithm.
A
preferred, non-limiting example of a mathematical algorithm utilized for the
comparison of
two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad.
Sci. U.S.A.
87:2264-2268, modified as in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci.
U.S.A.
90:5873-5877. Such an algorithm is incorporated into the NBLAST and XBLAST
programs
of Altschulet al., 1990, J. Mol. Biol. 215:403. BLAST nucleotide searches can
be performed
with the NBLAST nucleotide program parameters set e.g. for score=100,
word1ength=12 to
obtain nucleotide sequences homologous to a nucleic acid molecules of the
present invention.
BLAST protein searches can be performed with the XBLAST program parameters set
e.g. to
Score=50, wordlength=3 to obtain amino acid sequences homologous to a protein
molecule
of the present invention. To obtain gapped alignments for comparison purposes,
Gapped
BLAST can be utilized as described in Altschul et al., 1997, Nucleic Acids
Res. 25:3389-
3402. Alternatively, PSI-BLAST can be used to perform an iterated search which
detects
distant relationships between molecules. When utilizing BLAST, Gapped BLAST
and/or
PSI-Blast programs, the default parameters of the respective programs (e.g.,
of XBLAST and
NBLAST) can be used (see e.g. the NCBI website). Another preferred, non-
limiting example
of a mathematical algorithm utilized for the comparison of sequences is the
algorithm of
Myers and Miller, 1988, CABIOS 4:11-17. Such an algorithm is incorporated in
the ALIGN
program (version 2.0) which is part of the GCG sequence alignment software
package. When
utilizing the ALIGN program for comparing amino acid sequences, a PAM 120
weight
residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.
The percent
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identity between two sequences can be determined using techniques similar to
those
described above, with or without allowing gaps. In calculating percent
identity, typically only
exact matches are counted.
As used herein, the term "derivative" in the context of a proteinaceous agent
refers to a
proteinaceous agent that comprises an amino acid sequence which has been
altered by the
introduction of amino acid residue substitutions, deletions or additions. The
term "derivative'
as used herein also refers to a proteinaceous agent which, has been modified,
i.e. by the
covalent attachment of any type of molecule to the proteinaceous agent. For
example, but not
by way of limitation, a protein/antibody may be modified, e.g., by
glycosylation, acetylation,
pegylation, phosphorylation, amidation, derivatization by known
protecting/blocking groups,
proteolytic cleavage, linkage to a cellular ligand or other protein, etc. A
derivative of a
proteinaceous agent may be produced by chemical modifications using techniques
known to
those of skill in the art, including, but not limited to specific chemical
cleavage, acetylation,
formylation, etc. Further, a derivative of a proteinaceous agent may contain
one or more non-
classical amino acids. In a preferred embodiment, a proteinaceous derivative
possesses a
similar or identical function as the proteinaceous agent from which it was
derived.
Stereoisomers
All stereoisomers of Formula [X], such as those, for example, which may exist
due to
asymmetric carbons, including enantiomeric forms (which may exist even in the
absence of
asymmetric carbons) and diastereomeric forms, are contemplated and within the
scope of this
invention. Individual stereoisomers of the compounds of this invention may,
for example, be
substantially free of other isomers, or may be admixed, for example, as
racemates or with all
other or other selected, stereoisomers. The chiral centers of the present
invention can have the
S or R configuration as defined by the IUPAC 1974 Recommendations.
This disclosure encompasses the use of stereomerically pure forms of a
compound(s) of this
invention, as well as the use of mixtures of those forms. For example,
mixtures comprising
equal or unequal amounts of the enantiomers of a compound(s) of Formula [X],
for example
a compound(s) of Formula (I), may be used in methods and compositions
disclosed herein.
These isomers may be asymmetrically synthesized or resolved using standard
techniques
such as chiral columns or chiral resolving agents. See, e.g., Jacques, J., et
al., Enantiomers,
Racemates and Resolutions (Wiley-Interscience, New York, 1981); Wilen, S. H.,
et al.,
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Tetrahedron 33:2725 (1977); Eliel, E. L., Stereochemistry of Carbon Compounds
(McGraw-
Hill, N Y, 1962); and Wilen, S. H., Tables of Resolving Agents and Optical
Resolutions p.
268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind., 1972).
For the molecules presented in this invention's Description and Drawings: the
present
invention contemplates all polymorphs, crystals, co-crystals, metabolites,
isotopologues,
geometric/conformational isomers, rotamers, atropisomers, atropenantiomers,
stereoisomers,
optically active forms, racemates, scalemates, tautomers, keto-enol tautomers,
cis- and trans-
isomers, E and Z isomers, R- and S-enantiomers, diastereomers, isomers, (D)-
isomers, (L)-
.. isomers, the racemic mixtures thereof, other mixtures thereof and isotopic
variants (e.g.
deuterium in place of hydrogen in some or all places upon the molecule {s}) as
falling within
the scope of the invention. All such isomers, as well as mixtures thereof, are
intended to be
included in this invention. As well as analogues and
pharmaceutically/physiologically
acceptable salts/ethers/esters/solvates/hydrates/solvates of
salts/chelates/complexes/metal
complexes/mixtures/prodrugs/particles/radionuclides/derivatives/carriers/crysta
lline
forms/isomorphic crystalline forms/crystals/co-
crystals/clathrates/liposomes/nanoparticles/micronized forms/polymer
matrices/microspheres/vesicles/micelles/compositions/formulations/doses/combina
tionsN-
oxides/acid salt hydrates thereof. Unless indicated otherwise, chemical
structures and
graphical representations of compounds herein encompass all stereoisomers,
racemates,
scalemates, relative proportions/combinations of R and S stereoisomers and
optically active
or inactive forms. Optically active forms may be prepared by resolution of
racemic forms or
by synthesis from optically active starting materials. Many geometric isomers
of CC double
bonds, C=N double bonds, ring systems, and the like can also be present in the
compounds,
and all such stable isomers are contemplated in the present invention.
Substituents around a
carbon-carbon double bond are designated as being in the "Z" or "E"
configuration wherein
the terms "Z" and "E" are used in accordance with IUPAC standards. Unless
otherwise
specified, structures depicting double bonds encompass both the "E" and "Z"
isomers.
Compounds of the present invention, free form and salts thereof, may exist in
multiple
tautomeric forms, in which hydrogen atoms are transposed to other parts of the
molecules and
the chemical bonds between the atoms of the molecules are consequently
rearranged. It
should be understood that all tautomeric forms, insofar as they may exist, are
included within
the invention. The present invention is not limited to any particular
mechanism, nor to any
understanding of the action of the agents being administered.
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The invention also embraces isotopically labelled compounds of the invention
which are
identical to those recited herein, except that one or more atoms are replaced
by an atom
having an atomic mass or mass number different from the atomic mass or mass
number
usually found in nature. Isotopes include those atoms having the same atomic
number but
different mass numbers. By way of general example and without limitation,
examples of
isotopes that can be incorporated into compounds of the invention include
isotopes of
hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine,
such
as 2H, 3H, 13C, 14C, 15N, 180, 170, 31p, 32p, 35s, 18F, and
ui respectively. Isotopically
labelled compounds of the present disclosure can generally be prepared by
following
proceedures analogous to those disclosed herein by substituting an
isotopically labelled
reagent for a non-isotopically labelled reagent. Such isotopically labeled
compounds may be
useful in metabolic studies, reaction kinetic studies, detection or imaging
techniques, such as
positron emission tomography (PET) or single-photon emission computed
tomography
(SPECT) including drug or substrate tissue distribution assays or in
radioactive treatment of
subjects (e.g. humans). Substitution with positron emitting isotopes, such as
'IC, r '50 and
can be useful in PET studies for examining substrate receptor occupancy. These
radiolabelled compounds can be useful to further determine or measure the
effectiveness of
the compounds, by characterizing, for example, the site/mode of action, or
binding affinity to
pharmacologically important site of action. Substitution with heavier isotopes
such as
deuterium, i.e., 2H, may afford certain therapeutic advantages resulting from
greater
metabolic stability, for example, increased in vivo half-life or reduced
dosage requirements,
and hence may be preferred in some circumstances. Also provided for
isotopically labeled
compounds described herein are any pharmaceutically acceptable salts,
solvates, hydrates or
.. prodrugs, as the case may be.
In cases wherein there are nitrogen atoms (e.g., amines) on compounds of the
present
invention, these may be converted to N-oxides by treatment with an oxidizing
agent (e.g.,
mCPBA and/or hydrogen peroxides) to afford other compounds of this invention.
Thus, shown/claimed nitrogen atoms are considered to cover both the shown
nitrogen and its
N-oxide (N¨*0) derivative.
Any metabolite of a compound of this invention is a compound of this
invention.
Componentry to the invention is any compound of this invention, optionally a
compound of
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Formula [X], (I), (II), (III), (IV), (V), (VI), (VII) herein, with one or more
of its H
atoms/isotopes replaced with OH (hydroxyl), for example catalysed/enabled by
an enzyme
e.g. one or more of a Cytochorome P450 enzyme(s), monoxygenase enzyme(s),
hydroxylase
enzyme(s) and/or by other relevant enzyme(s) known to those of the art.
Optionally, but not
restrictively, such a compound(s) of the invention can be produced by
incubating a
compound(s) of the invention with microsomes, optionally liver microsomes,
wherein this
technique is well known to those of the art.
Reference to a compound(s) of Formula [X], (I), (II), (III), (IV), (V), (VI),
(VII) herein is
understood to include reference to every tautomer, enantiomer, mixture of
enantiomers, salt,
solvate, hydrate, prodrug, chemically protected form, ester, N-oxide,
metabolite, crystal,
polymorph, co-crystal, clathrate thereof, unless otherwise indicated.
Salts & solvates
Throughout the specification, groups and substituents thereof may be chosen by
one skilled in
the field to provide stable moieties and compounds.
The compounds of Formula [X] form salts which are also within the scope of
this invention.
Reference to a compound of the Formula [X] herein is understood to include
reference to
salts thereof, unless otherwise indicated. For illustrating, not restricting,
example: quaternary
ammonium salts of compounds of Formula [X] are componentry to the present
invention.
Generally speaking, salts tend to be more soluble in aqueous or other protonic
solvents than
are the corresponding free base forms.
The phrase "pharmaceutically acceptable" is employed herein to refer to those
compounds,
materials, compositions, and/or dosage forms that are, within the scope of
sound medical
judgment, suitable for use in contact with the tissues of human beings and
animals without
excessive toxicity, irritation, allergic response, and/or other problem or
complication,
commensurate with a reasonable benefit/risk ratio.
As used herein, the term "pharmaceutically acceptable salt" refers to any
pharmaceutically
acceptable salt (e.g., acid or base) of a compound of the present invention
which, upon
administration to a subject, is capable of providing a compound of this
invention or an active
metabolite or residue thereof. As is known to those of ordinary skill in the
art, "salts" of the
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compounds of the present invention may be derived from inorganic or organic
acids and
bases. For therapeutic use, salts of the compounds of the present invention
are contemplated
as being pharmaceutically acceptable (i.e., non-toxic, physiologically
acceptable). However,
salts of acids and bases that are non-pharmaceutically acceptable may also
find use, for
example, in the preparation, isolation or purification of a pharmaceutically
acceptable
compound. Lists of suitable salts are found in Remington's Pharmaceutical
Sciences, 18th
Edition, Mack Publishing Company, Easton, Pa. (1990) and Handbook of
Pharmaceutical
Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002)
and refer
Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19, all hereby
incorporated by
reference.
The term "salt(s)", as employed herein, denotes acidic and/or basic salts
formed with
inorganic and/or organic acids and bases. In addition, when a compound of
Formula [X]
contains both a basic moiety, such as, but not limited to an amine or a
pyridine or imidazole
ring, and an acidic moiety, such as, but not limited to a carboxylic acid,
zwitterions ("inner
salts") may be formed and are included within the term "salt(s)" as used
herein.
Salts can be prepared in situ during the final isolation and purification of
the compounds of
the invention, or by separately reacting a purified compound of the invention
in its free base
form with a suitable organic or inorganic acid, and isolating the salt thus
formed. Salts of the
compounds of the Formula [X] may be formed, for example, by reacting a
compound of the
Formula [X] with an amount of acid or base, such as an equivalent amount, in a
medium such
as one in which the salt precipitates or in an aqueous medium followed by
lyophilization.
The compounds of Formula [X] which contain a basic moiety, such as, but not
limited to an
amine or a pyridine or imidazole ring, may form salts with a variety of
organic and inorganic
acids. Exemplary acid addition salts include acetates (such as those formed
with acetic acid
or trihalo acetic acid, for example, trifluoroacetic acid), adipates,
alginates, ascorbates,
aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates,
citrates, camphorates,
camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfates,
ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates,
hemisulfates, heptanoates,
hexanoates, hydrochlorides (formed with hydrochloric acid), hydrobromides
(formed with
hydrogen bromide), hydroiodides, 2-hydroxyethanesulfonates, lactates, maleates
(formed
with maleic acid), methane-sulfonates (formed with methanesulfonic acid), 2-
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CA 3050553 2019-07-25
naphthalenesulfonates, nicotinates, nitrates, oxalates, pectinates,
persulfates, 3-
phenylpropionates, phosphates, picrates, pivalates, propionates, salicylates,
succinates,
sulfates (such as those formed with sulfuric acid), sulfonates (such as those
mentioned
herein), tartrates, thiocyanates, toluenesulfonates such as tosylates,
undecanoates, and the
like.
The compounds of Formula [X] which contain an acidic moiety, such as, but not
limited to a
carboxylic acid, may form salts with a variety of organic and inorganic bases.
Exemplary
basic salts include ammonium salts, alkali metal salts such as sodium,
lithium, and potassium
salts, alkaline earth metal salts such as calcium and magnesium salts, salts
with organic bases
(for example, organic amines) such as benzathines, dicyclohexylamines,
hydrabamines
[formed with N,N-bis(dehydro-abietyl)ethylenediamine], N-methyl D-glucamines,
N-methyl-
D-glucamides, t-butyl amines, and salts with amino acids such as arginine,
lysine and the
like. Basic nitrogen-containing groups may be quaternized with agents such as
lower alkyl
halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides and
iodides), dialkyl
sulfates (e.g., dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain
halides (e. g., decyl,
lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides
(e.g., benzyl and
phenethyl bromides), and others.
Compounds of Formula [X], and salts thereof, may exist in their tautomeric
form (for
example, as an amide or imino ether). All such tautomeric forms are
contemplated herein as
part of the present invention.
It should be understood that solvates (e.g., hydrates) of a compound(s) of
Formula [X] are
also within the scope of the present invention. Herein, the term "solvate"
means a compound
provided herein or a salt thereof that further includes a stoichiometric or
non-stoichiometric
amount of solvent (organic or inorganic) bound by non-covalent intermolecular
forces (e.g.
hydrogen bonding). Where the solvent is water, the solvate is a hydrate. In
certain instances
the solvate will be capable of isolation, for example when one or more solvent
molecules are
incorporated in the crystal lattice of the crystalline solid. The solvent
molecules in the solvate
may be present in a regular arrangement and/or a non-ordered arrangement. The
solvate may
comprise either a stoichiometric or nonstoichiometric amount of the solvent
molecules.
"Solvate" encompasses both soluiion-phase and isolable solvates. Exemplary
solvates
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include, but are not limited to, hydrates, ethanolates, methanolates, and
isopropanolates.
Methods of solvation are known in the art.
Chelates, metal complexes (metal complexes include calcium, zinc, iron and the
like),
mixtures, radionuclides and liposomes of Formula [X] are within the scope of
this invention.
In some embodiments, a compound(s) of Formula [X] is anhydrous.
Fatty acids
In an embodiment, a therapeutic compound(s) componentry to the present
invention, for
example a compound(s) of Formula [X], is formulated with a pharmaceutically-
acceptable
fatty acid(s). In an embodiment, the stoichiometry of a compound of Formula
[X] to a fatty
acid is 1:1, in another embodiment it is 1:2, and in other embodiments it is
other
stoichiometries/ratios. What constitutes a fatty acid is well known to those
of the art, and isn't
necessarily limited to the brief specification given herein. A fatty acid is a
carboxylic acid
with an aliphatic chain, which is either saturated (no C=C double bonds; in
some cases with a
formula CH3(CH2)nCOOH where n is a positive integer) or unsaturated (with one
or more
C=C double bonds, each of which can be in a cis or trans configuration;
monounsaturated
fatty acids have a single C=C double bond, polyunsaturated fatty acids (PUFA)
have more
than one C=C double bond, methylene-interrupted polyenes have two or more cis
double
bonds separated from each other by a single methylene bridge (-CH2-) (i.e. C=C-
C-C=C),
conjugated fatty acids have at least one pair of double bonds separated by
only a single bond
(i.e. C=C-C=C)), which is either branched or (more typically) unbranched,
which is
optionally substituted. This arrangement confers a fatty acid with a polar,
hydrophilic end,
and a nonpolar, hydrophobic end that is insoluble in water. Most naturally
occurring fatty
acids have an unbranched hydrocarbon chain of an even number of carbon atoms,
typically
between 4 and 28 carbons, and may be attached to functional groups containing
oxygen,
halogens, nitrogen, and sulfur. Synthetic or non-natural fatty acids may have
a hydrocarbon
chain of any number of carbon atoms from between 3 and 40 carbons. Where a
double bond
exists, there is the possibility of either cis or trans geometric isomerism (=
EIZ isomerism in
IUPAC nomenclature), which significantly affects the molecule's molecular
configuration.
Cis-double bonds cause the fatty acid chain to bend, an effect that is more
pronounced the
more cis double bonds there are in a chain. Most naturally occurring fatty
acids are of the cis
configuration, although the trans form does exist in some natural and
partially hydrogenated
fats and oils, wherein trans fats have been associated with increased risk of
coronary heart
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disease. Omega (co) is the name for the methyl end of a fatty acid and,
starting from the
omega end, omega-3 (co-3 or n-3) fatty acids have their first (in some cases
only) double bond
between the 3rd and zith carbon atoms from the methyl end, omega-6 (co-6 or n-
6) fatty acids
have their first (in some cases only) double bond between the 6th and 7th
carbon atoms from
.. the methyl end, omega-7 (co-7 or n-7) fatty acids have their first (in some
cases only) double
bond between the 71h and 8th carbon atoms from the methyl end, omega-9 (co-9
or n-9) fatty
acids have their first (in some cases only) double bond between the 9th and
10th carbon atoms
from the methyl end. In naming fatty acids a form used is, to illustrate with
an example,
(18:3, n-3), wherein this fatty acid has 18 carbons in its aliphatic chain, 3
double bonds, the
first of which emanates from the 3rd carbon from the methyl (omega, o)) end of
the chain (i.e.
this is an omega-3 fatty acid). To give a further example, (20:5, n-3),
wherein this fatty acid
has 20 carbons in its aliphatic chain, 5 double bonds, the first of which
emanates from the 3'
carbon from the methyl (omega, co) end of the chain (i.e. this is an omega-3
fatty acid). To
give yet a further example, (20:3, n-9), wherein this fatty acid has 20
carbons in its aliphatic
chain, 3 double bonds, the first of which emanates from the 9th carbon from
the methyl
(omega, co) end of the chain (i.e. this is an omega-9 fatty acid). Note that
these examples
haven't been fully specified using this notation because the cis/trans (EIZ)
isomerism of their
double bond(s) hasn't been specified. Also, note that this naming system is
not what is
specified by IUPAC, where for one thing, carbons are counted from the carboxyl
rather than
the methyl end of the aliphatic chain. a-Linolenic acid (ALA) is (18:3, n-3)
and in IUPAC
nomenclature: (9Z,12Z,15Z)-9,12,15-Octadecatrienoic acid, wherein this fatty
acid has 3
double bonds, all Z configuration, at carbons 9, 12, 15 from the carboxyl end,
wherein (to
revert back away from IUPAC) its first double bond emanates from the 3rd
carbon atom from
the methyl end.
Examples of fatty acids include, without limitation, Capryllic acid, Caprylic
acid, pelargonic
acid, Capric acid. Undecylic acid, Lauric acid, Tridecylic acid, Myristic
acid, Myristoleic
acid, Pentadecyclic acid, Palmitic acid, Palmitoleic acid, Sapienic acid,
Margaric acid, Stearic
acid, Oleic acid, Elaidic acid, Vaccenic acid, Linoleic acid, Linoelaidic
acid, a -Linolenic
acid, y -Linolenic acid, Stearidonic acid, Nonadecylic acid, Arachidic acid,
Eicosenoic acid,
11-Eicosenoic acid, Dihomo-y-linolenic acid, Mead acid, Arachidonic acid,
Eicosapentaenoic
acid, Heneicosylic acid, Behenic acid, Erucic acid, Docosahexaenoic acid,
Tricosylic acid,
Lignoceric acid, Nervonic acid, Pentacosylic acid, Cerotic acid, Heptacosylic
acid, Montanic
acid, Nonacosylic acid, Melissic acid, Henatriacontylic acid, Lacceroic acid,
Psyllic acid,
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Geddic acid, Ceroplastic acid, and Hexatriacontylic acid, a-Linolenic acid, y-
Linolenic acid,
omega 3 fatty acid (ALA), omega 3 fatty acid (DHA), omega 3 fatty acid (EPA),
omega 6
fatty acid (AA), omega 7 fatty acid (CLA), omega 9 fatty acid (OA), omega 9
hydroxylated
fatty acid (RA), Arachidonic acid, [9Z, 11E conjugated Linoleic acid],
conjugated linoleic
acid, conjugated (9Z,11E)-Linoleic acid (CAS Number 2540-56-9),
Docosahexaenoic acid,
Eicosapentaenoic acid, Oleic acid, Ricinoleic acid, butyric acid, hexanoic
acid, decanoic acid,
valeric acid, pentadecan acid, heptanoic acid, docosatetraenoic acid,
heptadecanoic acid,
paullinic acid, alpha-parinaric acid, calendic acid, docosapentaenoic acid,
linolelaidic acid,
gadoleic acid, alpha-Eleostearic acid, petroselinic acid, punicic acid,
pinolenic acid, rumenic
acid, eicosatetraenoic acid, tuberculostearic acid, vernolic acid, prostanoic
acid, catalpic acid,
jacaric acid, malvalic acid, aleuritic acid, lesquerolic acid,
bosseopentaenoic acid,
hydnocarpic acid.
In aspects of this embodiment, a saturated or unsaturated fatty acid
comprises, e.g., at least 8,
at least 10, at least 12, at least 14, at least 16, at least 18, at least 20,
at least 22, at least
24, at least 26, at least 28, or at least 30 carbon atoms. In other aspects of
this embodiment, a
saturated or unsaturated fatty acid comprises, e.g., between 4 and 24 carbon
atoms, between
6 and 24 carbon atoms, between 8 and 24 carbon atoms, between 10 and 24 carbon
atoms,
between 12 and 24 carbon atoms, between 14 and 24 carbon atoms, or between 16
and
24 carbonatoms, between 4 and 22 carbonatoms, between 6 and 22 carbon atoms,
between 8
and 22 carbon atoms, between 10 and 22 carbon atoms, between 12 and 22 carbon
atoms,
between 14 and 22 carbon atoms, or between 16 and 22 carbonatoms, between 4
and 20
carbon atoms, between 6 and 20 carbon atoms, between 8 and 20 carbon atoms,
between 10
and 20 carbon atoms, between 12 and 20 carbon atoms, between 14 and 20 carbon
atoms, or
between 16 and 20 carbon atoms. If unsaturated, the fatty acid may have, e.g.,
1 or more, 2 or
more, 3 or more, 4 or more, 5 or more, or 6 or more double bonds.
In another embodiment, an adjuvant may comprise one kind of pharmaceutically-
acceptable
fatty acid. In another embodiment, an adjuvant may comprise a plurality of
different
.. pharmaceutically-acceptable fatty acids. In aspects of this embodiment, an
adjuvant may
comprise, e.g., two or more different fatty acids, three or more different
fatty acids, four or
more different fatty acids, five or more different fatty acids, or six or more
different fatty
acids.
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A pharmaceutically-acceptable fatty acid useful in the pharmaceutical
compositions disclosed
herein may be a pharmaceutically-acceptable omega fatty acid. Non-limiting
examples of an
omega fatty acid include an omega-3 fatty acid, an omega-6 fatty acid, an
omega-7 fatty acid,
an omega-9 fatty acid.
Omega-3 fatty acids (also known as n-3 fatty acids or a)-3 fatty acids) are a
family of
unsaturated fatty acids that have in common a final carbon-carbon double bond
in the n-3
position, that is, the third bond, counting from the methyl end of the fatty
acid. Omega-3 fatty
acids are "essential" fatty acids because they are vital for normal metabolism
and cannot be
synthesized by the human body. An omega-3 fatty acid includes, without
limitation,
Hexadecatrienoic acid (16:3), a-Linolenic acid (18:3), Stearidonic acid
(18:4), Eicosatrienoic
acid (20:3), Eicosatetraenoic acid (20:4), Eicosapentaenoic acid (20:5),
Heneicosapentaenoic
acid (21:5), Docosapentaenoic acid (Clupanodonic acid) (22:5), Docosahexaenoic
acid
(22:6), Tetracosapentaenoic acid (24:5), Tetracosahexaenoic acid
(Nisinic acid) (24:6).
Omega-6 fatty acids (also known as n-6 fatty acids or co-6 fatty acids) are a
family of
unsaturated fatty acids that have in common a final carbon-carbon double bond
in the n-6
position, that is, the sixth bond, counting from the methyl end of the fatty
acid. An omega-6
fatty acid includes, without limitation, Linoleic acid (18:2), y-linolenic
acid (18:3), Calendic
acid (18:3), Eicosadienoic acid (20:2), Dihomo-y-linolenic acid (20:3),
Arachidonic acid
(20:4), Docosadienoic acid (22:2), Adrenic acid (22:4). Docosapentaenoic acid
(22:5),
Tetracosatetraenoic acid (24:4), and Tetracosapentaenoic acid (24:5).
Omega-7 fatty acids (also known as n-7 fatty acids or a)-7 fatty acids) are a
family of
unsaturated fatty acids that have in common a final carbon-carbon double bond
in the n-7
position, that is, the seventh bond, counting from the methyl end of the fatty
acid. An omega-
7 fatty acid includes, without limitation, 5-Dodecenoic acid (12:1), 7-
Tetradecenoic acid
(14:1), 9-1-lexadecenoic acid (Palmitoleic acid) (16:1), 11-Decenoic acid
(Vaccenic acid)
(18:1), 9Z, 11E conjugated Linoleic acid (Rumenic acid) (18:2), 13-Eicosenoic
acid
(Paullinic acid) (20:1), 15-Docosenoic acid (22:1), and 17-Tetracosenoic acid
(24:1).
Omega-9 fatty acids (also known as n-9 fatty acids or 6)-9 fatty acids) are a
family of
unsaturated fatty acids that have in common a final carbon-carbon double bond
in the n-9
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position, that is, the ninth bond, counting from the methyl end of the fatty
acid. An omega-9
fatty acid includes, without limitation, Oleic acid (18:1), Elaidic acid
(18:1), Eicosenoic acid
(20:1), Mead acid (20:3), Erucic acid (22:1), Nervonic acid (24:1), Ricinoleic
acid, ximenic
acid (26:1).
A pharmaceutically-acceptable fatty acid useful in the pharmaceutical
compositions disclosed
herein may be a pharmaceutically-acceptable conjugated fatty acid. Conjugated
fatty acids
are positional and geometric isomers of polyunsaturated fatty acids in which
at least one pair
of double bonds are separated by only one single bond. In one aspect of this
embodiment, a
pharmaceutically-acceptable conjugated fatty acid is, e.g., a C16 conjugated
fatty acid, a C18
conjugated fatty acid, a C20 conjugated fatty acid, a C22 conjugated fatty
acid, a C24
conjugated fatty acid, a C26 conjugated fatty acid, a C28 conjugated fatty
acid or a C30
conjugated fatty acid. In one aspect of this embodiment, pharmaceutically-
acceptable
conjugated fatty acid is, e.g., a C16-C18 conjugated fatty acid, a C 16-C20
conjugated fatty
acid, a C16-C22 conjugated fatty acid, a C16-C24 conjugated fatty acid, a C16-
C26
conjugated fatty acid, a C16-C28 conjugated fatty acid, a C16-C30 conjugated
fatty acid, a
C18-C20 conjugated fatty acid, a C18-C22 conjugated fatty acid, a C 18-C24
conjugated fatty
acid, a C18-C26 conjugated fatty acid, a C18-C28 conjugated fatty acid, a C18-
C30
conjugated fatty acid, a C20-C22 conjugated fatty acid, a C20-C24 conjugated
fatty acid, a
C20-C26 conjugated fatty acid, a C20-C28 conjugated fatty acid, a C20-C30
conjugated fatty
acid, a C22-C24 conjugated fatty acid, a C22-C26 conjugated fatty acid, a C22-
C28
conjugated fatty acid, a C22-C30 conjugated fatty acid, a C24-C26 conjugated
fatty acid, a
C24-C28 conjugated fatty acid, a C24-C30 conjugated fatty acid, a C26-C28
conjugated fatty
acid, a C26-C30 conjugated fatty acid, or C28-C30 conjugated fatty acid.
In another aspect of this embodiment, a pharmaceutically acceptable conjugated
fatty acid
includes, e.g., a conjugated Linoleic acid, a conjugated Linoelaidic acid, a
conjugated a-
Linolenic acid, a conjugated y-Linolenic acid, a conjugated Calendic acid, a
conjugated
Eicosadienoic acid, a conjugated Stearidonic acid, a conjugated Nonadecylic
acid, a
conjugated Arachidic acid, a conjugated Dihomo-y-linolenic acid, a conjugated
DocoSadienoic, a conjugated Mead acid, a conjugated Arachidonic acid, a
conjugated
Eicosapentaenoic acid, a conjugated Adrenic acid, a conjugated
Docosapentaenoic acid, a
conjugated Heneicosylic acid, a conjugated Tetracosatetraenoic acid, a
conjugated
Tetracosapentaenoic acid, a conjugated Behenic acid, a conjugated
Docosahexaenoic acid, a
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conjugated Tricosylic acid, a conjugated Lignoceric acid, a conjugated
Pentacosylic acid, a
conjugated Cerotic acid, a conjugated Heptacosylic acid, a conjugated Montanic
acid, a
conjugated Nonacosylic acid, a conjugated Melissic acid, a conjugated
Henatriacontylic acid,
a conjugated Lacceroic acid, a conjugated Psyllic acid, a conjugated Geddic
acid, a
conjugated Ceroplastic acid, a conjugated Hexatriacontylic acid, or any
combination thereof.
A pharmaceutically-acceptable fatty acid useful in the pharmaceutical
compositions disclosed
herein may be a pharmaceutically-acceptable conjugated linoleic acid (CLA).
Conjugated
linoleic acid (CLA) refers to a group of at least 28 positional and geometric
isomers of the
omega-6 essential fatty acid linoleic acid (cis-9, cis-12, octadecadienoic
acid). The double
bonds of CLAS are conjugated, with only one single bond between them.
Virtually all cis-
and trans-isomeric combinations of CLA have been identified. A CLA includes,
without
limitation, cis-9, trans-11, octadecadienoic acid (c-9, t-11 CLA), cis-9, cis-
11,
octadecadienoic acid (c-9, c-11 CLA), trans-9, trans-11, octadecadienoic acid
(t-9, t-11
CLA), and trans-9, cis-11, octadecadienoic acid (t-9, c-11 CLA), cis-9, trans-
11, conjugated
linoleic acid (c-9, t-11 CLA), cis-9, cis 11, conjugated linoleic acid (c-9, c-
11 CLA), trans-9,
trans 11, conjugated linoleic acid (t-9, t-11 CLA), and trans-9, cis-11,
conjugated linoleic acid
(t-9, c-11 CLA), cis-10, trans 12, conjugated linoleic acid (c-10, t-12 CLA),
cis-10, cis-12,
conjugated linoleic acid (c-10, c-12 CLA), trans-10, trans-12, conjugated
linoleic acid (t-10,
t-12 CLA), and trans-i0, cis 12, conjugated linoleic acid (t-10, c-12 CLA),
and any
combination thereof.
In an aspect of this embodiment, a pharmaceutical composition comprises a
compound(s) of
Formula [X] and an Omega-3 fatty acid(s). In another aspect of this
embodiment, a
pharmaceutical composition comprises a compound(s) of Formula [X] and an Omega-
6 fatty
acid(s). In yet another aspect of this embodiment, a pharmaceutical
composition comprises a
compound(s) of Formula [X] and an Omega-7 fatty acid(s). In still another
aspect of this
embodiment, a pharmaceutical composition comprises a compound(s) of Formula
[X] and an
Omega-9 fatty acid(s). In other aspects, a pharmaceutical composition
comprises a
compound(s) of Formula [X] and an Omega-3 fatty acid, an Omega-6 fatty acid,
an Omega-7
fatty acid, an Omega-9 fatty acid, or any combination thereof. In yet other
aspects, a
pharmaceutical composition comprises a compound(s) of Formula [X] and C-
Linolenic acid,
Arachidonic acid, Docosahexaenoic acid, Rumenic acid, or any combination
thereof.
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In an embodiment, a pharmaceutical composition comprises one or more compounds
of
Formula [X] and one or more of a fatty acid(s), wherein the fatty acid is an
omega-3 fatty
acid, an omega-6 fatty acid, an omega-7 fatty acid, an omega-9 fatty acid, or
any combination
thereof,
wherein (if present) the omega-3 fatty acid is Hexadecatrienoic acid (16:3), C-
Linolenic acid
(18:3), Stearidonic acid (18:4), Eicosa trienoic acid (20:3), Eicosatetraenoic
acid (20:4),
Eicosa pentaenoic acid (20:5), Heneicosapentaenoic acid (21:5),
Docosapentaenoic acid
(Clupanodonic acid) (22:5), Docosahexaenoic acid (22:6), Tetracosapentaenoic
acid (24:5),
Tetracosahexaenoic acid (Nisinic acid) (24:6), or any combination thereof,
wherein (if present) the omega-6 fatty acid is Linoleic acid (18:2), Y-
linolenic acid (18:3),
Calendic acid (18:3), Eicosadienoic acid (20:2), Dihomo-y-linolenic acid
(20:3), Arachidonic
acid (20:4), Docosadienoic acid (22:2), Adrenic acid (22:4).Docosapentaenoic
acid (22:5),
Tetracosatetraenoic acid (24:4), and Tetracosapentaenoic acid (24:5), or any
combination
thereof,
wherein (if present) the omega-7 fatty acid is 5-Dodecenoic acid, 7-
Tetradecenoic acid, 9-
Hexadecenoic acid (Palmitoleic acid), 11-Decenoic acid (Vaccenic acid), 13-
Eicosenoic acid
(Paullinic acid), 15-Docosenoic acid, 17-Tetracosenoic acid, and 9Z, 11E
conjugated Linoleic
acid (Rumenic acid), or any combination thereof,
wherein (if present) the omega-9 fatty acid Oleic acid, Elaidic acid,
Eicosenoic acid,
Meadacid, Erucic acid, Nervonic acid, and Ricinoleic acid, or any combination
thereof.
In an embodiment, a pharmaceutical composition comprises one or more compounds
of
Formula [X] and one or more of a fatty acid, plus optionally a
chemotherapeutic and/or anti-
proliferative agent(s) and/or one or more compounds approved for human use,
optionally for
anti-cancer use, by the United States Food and Drug Administration (FDA)
and/or European
Medicines Agency (EMA). Optionally, further/alternative constituent(s) are
componentry to
this composition, wherein the range of options is clear to someone of the art,
some of which
are disclosed elsewhere herein, and include (without limitation)
pharmaceutically-acceptable
carriers include vehicles, stabilizers, diluents, additives, auxiliarys or
excipients, including
buffers, preservatives, tonicity adjusters, salts, demulcents, antioxidants,
osmolality adjusting
agents, physiological substances, pharmacological substances, bulking agents,
emulsifying
agents, wetting agents, flavoring agents, or coloring agents, a
therapeutically effective
amount of a chemotherapeutic or anti-proliferative agent(s), wherein the
chemotherapeutic or
anti-proliferative agent(s) is selected from the group consisting of an
alkylating agent, a
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platinum agent, an antimetabolite, a topoisomerase inhibitor, an antitumor
antibiotic, an
aromatase inhibitor, a thymidylate synthase inhibitor, a DNA antagonist,
farnesyltransferase
inhibitor, a pump inhibitor, a metalloproteinase inhibitor, a ribonucleoside
reductase
inhibitor, a TNFC, agonist, an endothelin A receptor antagonist, a retinoic
acid
receptoragonist, an immuno-modulator, a hormonal and antihormonal agent, a
photodynamic
agent, a tyrosine kinase inhibitor, and any combination thereof.
An aspect of the invention is to use a composition(s) componentry to this
disclosure, for
(non-limiting) example a composition containing a compound(s) of Formula [X]
and a fatty
acid(s), in a method of treatment of the human or animal body by therapy, for
(non-limiting)
example to treat/ameliorate/prevent/combat one or more of the
diseases/disorders/pathologies/damages/processes mentioned herein, including
(non-limiting)
aging, signs of aging, diseases of aging, cancer and cachexia. A yet further
aspect is to use a
composition componentry to this disclosure, for (non-limiting) example a
composition
containing a compound(s) of Formula [X] and a fatty acid(s), for the
manufacture of a
medicament for the treatment/amelioration/prevention/combat of body damage,
aging and/or
a disease/disorder/pathology and/or to enhance/improve body/brain function,
optionally for
one or more of the therapeutic/beneficial/desirable applications mentioned
herein, optionally
cancer.
Fatty acids can have anti-cancer activity (e.g. refer [303, 304]). In an
embodiment, when
formulating a composition of a compound(s) of Formula [X] and a fatty acid(s),
if this
composition is to be used for an anti-cancer use, the fatty acid(s) selected
should, in a
preferred embodiment, be a fatty acid(s) with the greatest anti-cancer
activity, especially the
fatty acid(s) with the greatest anti-cancer activity at the dosage used. In
especially preferred
composition embodiments the anti-cancer activity of a compound(s) of Formula
[X] and a
fatty acid(s) synergise.
An invention embodiment is a pharmaceutical composition comprising a
therapeutically
effective amount of a compound(s) of Formula [X] and 9Z,I1E conjugated
linoleic acid,
optionally in a 1:1 ratio.
Prod rugs
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In addition, compounds of the Formulas [X] may have prodrug forms. Any
compound that
will be converted or react under biological conditions, e.g. in vivo, to
provide the bioactive
agent (i.e., a compound of Formula [X]) is a prodrug within the scope and
spirit of the
invention. For example, a derivative of a compound of Formula [X] that can
hydrolyze,
oxidize, or otherwise react under biological conditions to provide a compound
of Formula
[X]. For example, prodrug compounds of Formula [X] may be carboxylate ester
moieties. A
carboxylate ester may be conveniently formed by esterifying any of the
carboxylic acid
functionalities found on the disclosed structure(s). For example, prodrug
compounds of
Formula [X] comprise biohydrolyzable moieties such as biohydrolyzable amides,
biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable
carbonates,
biohydrolyzable ureides, and biohydrolyzable phosphate analogues. Other
examples of
prodrugs include derivatives of a compound of Formula [X] that comprise ¨NO,
¨NO2, ¨
ONO, or ¨0NO2 moieties. Various forms of prodrugs are well known in the art.
For
examples of such prodrug derivatives, see:
a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985), and Methods
in
Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et. al. (Academic Press,
1985);
b) A Textbook of Drug Design and Development, edited by Krosgaard-Larsen and
H.
Bundgaard, Chapter 5, "Design and Application of Prodrugs," by H. Bundgaard,
p. 113-191
(1991);
c) H. Bundgaard, Advanced Drug Delivery Reviews, Vol. 8, p. 1-38 (1992);
d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, Vol. 77, p. 285
(1988); and
e) N. Kakeya, et. al., Chem Phar Bull, Vol. 32, p. 692 (1984).
0 Burger's Medicinal Chemistry and Drug Discovery, 172-178, 949-982 (Manfred
E. Wolff
.. ed., 5th ed. 1995)
g) Medicinal Chemistry: Principles and Practice, King, F. D., ed. The Royal
Society of
Chemistry, Cambridge, UK (1994); Testa, B. et al., Hydrolysis in Drug and
Prodrug
Metabolism. Chemistry, Biochemistry and Enzymology, VCHA and Wiley-VCH,
Zurich,
Switzerland (2003); The Practice of Medicinal Chemistry, Wermuth, C. G., ed.,
Academic
Press, San Diego, Calif. (1999); Pro-drugs as Novel Delivery Systems, Vol. 14,
ACS
Symposium Series (T. Higuchi and W. Stella) and Bioreversible Carriers in Drug
Design,
Pergamon Press, 1987 (Ed. E. B. Roche, American Pharmaceutical Association).
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Prodrugs in accordance with the invention can, for example, be produced by
replacing
appropriate fimctionalities present in the compounds of Formula [X] with
certain moieties
known to those skilled in the art as 'pro-moieties' as described, for example,
in Design of
Prodrugs by H. Bundgaard (Elsevier, 1985), or in Prodrugs: Challenges and
Reward, 2007
edition, edited by Valentino Stella, Ronald Borchardt, Michael Hageman, Reza
Oliyai, Hans
Maag, Jefferson Tilley, pages 134-175 (Springer, 2007).
Moreover, certain compounds of Formula [X] may themselves act as prodrugs of
other
compounds of Formula (I).
Also included within the scope of the invention are metabolites of compounds
of Formula
[X], that is, compounds formed in vivo upon administration of the drug.
As used herein and unless otherwise indicated, the terms "biohydrolyzable
amide,"
"biohydrolyzable ester," "biohydrolyzable carbamate," "biohydrolyzable
carbonate,"
"biohydrolyzable ureide," and "biohydrolyzable phosphate" mean an amide,
ester, carbamate,
carbonate, ureide, or phosphate, respectively, of a compound that either: 1)
does not interfere
with the biological activity of the compound but can confer upon that compound
advantageous properties in vivo, such as uptake, duration of action, or onset
of action; or 2) is
biologically inactive but is converted in vivo to the biologically active
compound. Examples
of biohydrolyzable esters include, but are not limited to, lower alkyl esters,
lower
acyloxyalkyl esters (such as acetoxylmethyl, acetoxyethyl,
aminocarbonyloxymethyl,
pivaloyloxymethyl, and pivaloyloxyethyl esters), lactonyl esters (such as
phthalidyl and
thiophthalidyl esters), lower alkoxyacyloxyalkyl esters (such as
methoxycarbonyl-oxymethyl,
ethoxycarbonyloxyethyl and isopropoxycarbonyloxyethyl esters), alkoxyalkyl
esters, choline
esters, and acylamino alkyl esters (such as acetamidomethyl esters). Examples
of
.. biohydrolyzable amides include, but are not limited to, lower alkyl amides,
a-amino acid
amides, alkoxyacyl amides, and alkylaminoalkylcarbonyl amides. Examples of
biohydrolyzable carbamates include, but are not limited to, lower alkylamines,
substituted
ethylenediamines, amino acids, hydroxyalkylamines, heterocyclic and
heteroaromatic
amines, and polyether amines.
The term "prodrug" is also meant to include any covalently bonded carriers,
which release
the active compound of the invention in vivo when such prodrug is administered
to a
mammalian subject. Prodrugs of a compound of the invention may be prepared by
modifying
functional groups present in the compound of the invention in such a way that
the
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modifications are cleaved, either in routine manipulation or in vivo, to the
parent compound
of the invention. Prodrugs include compounds of the invention wherein a
hydroxy, amino or
mercapto group is bonded to any group that, when the prodrug of the compound
of the
invention is administered to a mammalian subject, cleaves to form a free
hydroxy, free amino
or free mercapto group, respectively. Examples of prodrugs include, but are
not limited to,
acetate, formate and benzoate derivatives of alcohol or amide derivatives of
amine functional
groups in the compounds of the invention and the like.
Dosage
As used herein, the term "therapeutically effective amount" or "effective
amount" refers to
the amount of an administered compound sufficient to effect a beneficial or
desired result(s)
in the subject, e.g. prevents, reduces or eliminates a cause and/or symptom(s)
of a
disease/disorder, optionally in combination with other active compound(s). An
effective
amount can be administered in one or more administrations, applications or
dosages and is
not intended to be limited to a particular formulation or administration
route.
The effective amount of a compound of the present invention may be determined
and by one
of ordinary skill in the art. The specific dose level and frequency of dosage
for any particular
subject may vary and will depend upon a variety of factors, including the
activity of the
specific compound employed, the metabolic stability and length of action of
that compound,
the pharmacokinetics of the compound, the formulation of compound used, the
species, age,
body weight, general health, medical condition, medical history, resilience,
sex and diet of
the subject, the mode/route and frequency of administration, rate of
excretion, renal and
hepatic function of patient, drug combination, concurrent treatment and type
and
severity/extent of the particular condition, nature/type/extent of presenting
symptoms, the
desired effect/outcome and/or the responsiveness/reaction of the subject. In
an embodiment, a
physician or veterinarian determines and prescribes the effective amount of
the drug required.
Broadly, small dosages may be used initially and, if necessary, increased by
small increments
until the desired effect under the circumstances is reached.
An exemplary effective amount of compounds of Formula [X] may be within the
dosage
range of about 0.001 to about 300 mg/kg, preferably about 0.2 to about 50
mg/kg and more
preferably about 0.5 to about 25 mg/kg (or from about 1 to about 2500 mg,
preferably from
about 5 to about 2000 mg) on a regimen in single or 2 to 6 (or more) divided
daily doses. But
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more exactly it depends upon the compound used, the condition and its
advancement/severity
(e.g. the type and grade of cancer), the route of administration, type of
dosing (e.g. pulse or
consistent etc.), what other treatments are undertaken alongside or previously
(e.g.
chemotherapeutics, surgery, radiotherapy, immunotherapy etc.), the age, sex,
condition,
previous/other diseases of the patient, pharmacokinetics of compound in that
patient,
response to treatment and exceptions to this dosage range are contemplated by
the present
invention, and can be changed during treatment to find the optimum. Optimal
dosage to be
administered to a subject can be determined by those skilled in the art.
Clinical trials may be
used to optimize the dose and dosing frequency for any particular compound,
with
subsequent further optimization for each particular subject by the direction
of one of ordinary
skill of the art e.g. a medical/veterinary practitioner. When the compounds
described herein
are co-administered with another agent, the effective amount may be less than
when the agent
is used alone.
Once improvement of the subject's disease/disorder/condition has occurred, the
dose may be
adjusted for preventative or maintenance treatment. For example, the dosage or
the frequency
of administration, or both, may be reduced as a function of the symptoms, to a
level at which
the desired therapeutic or prophylactic effect is maintained. Of course, if
symptoms have
been alleviated to an appropriate level, treatment may cease. Subject may,
however, require
intermittent treatment on a long-term basis upon any recurrence of symptoms.
Subjects may
also require chronic treatment on a long-term basis.
Pharmaceutical composition
While it is possible for a compound of the present invention to be
administered alone, it is
preferable to administer the compound as a pharmaceutical
formulation/composition. The
compound(s) of Formula [X], or an enantiomer or a mixture of enantiomers
thereof; or a
pharmaceutically acceptable salt, solvate, hydrate, prodrug, co-crystal,
clathrate, or
polymorph thereof, may be formulated, alone or together, in suitable dosage
unit with
pharmaceutically acceptable excipients, carriers, adjuvants and vehicles,
appropriate for each
route of administration.
The term "pharmaceutical composition" means a composition comprising a
compound of the
invention in combination with at least one additional pharmaceutically
acceptable carrier, ,
inert or active, making the composition especially suitable for diagnostic or
therapeutic use in
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vivo. Disclosed is a pharmaceutical composition of a therapeutically effective
amount of a
compound(s) of Formula [X] or a pharmaceutically acceptable salt thereof, and
one or more
pharmaceutically acceptable carriers, additives and/or diluents. This can be
manufactured by
a method of the art including, but not restricted to, mixing, dissolving,
granulating, dragee-
making, levigating, emulsifying, encapsulating, entrapping, melt-spinning,
spray-drying, or
lyophilizing processes. Pharmaceutical compositions may be presented in unit
dose forms
containing a predetermined amount of active ingredient, e.g. a compound(s) of
Formula [X],
per unit dose.
A "pharmaceutically acceptable carrier" refers to media generally accepted in
the art for the
delivery of biologically active agents to animals, in particular, mammals,
including (without
limitation), i.e., adjuvant, excipient, carrier or vehicle, such as diluents,
preserving agents,
preservatives, stabalizers, fillers, flow regulating agents, disintegrating
agents, encapsulating
materials, coating agents, release agents, wetting agents, emulsifying agents,
water,
phosphate buffered saline solution, emulsions (e.g., such as an oil/water or
water/oil
emulsions), suspending agents, anti-oxidants, buffers, pH buffers, tonicity
adjusters,
osmolality adjusting agents, physiological substances, pharmacological
substances,
sweetening agents, flavoring agents, coloring agents, perfuming agents,
bulking agents,
antibacterial agents, antifungal agents, surfactants, humectants, absorbents,
precipitation
inhibitors, adsorbants, solution retarding agents, solvents, anti-foaming
agents, salivary
stimulating agents, absorption accelerators, cooling agents, lubricating
agents, viscosity
enhancing agents, dispensing agents, and the like, depending on the nature of
the mode of
administration and dosage forms. Most preferably, but not restrictively, the
chosen
pharmaceutically acceptable vehicle(s) (e.g., carrier(s), adjuvant(s), and/or
other excipient(s))
has met the required standards of toxicological and manufacturing testing
and/or is included
on the Inactive Ingredient Guide prepared by the U.S. Food and Drug
administration.
Pharmaceutically acceptable carriers are formulated according to a number of
factors well
within the purview of those of ordinary skill in the art. These include,
without limitation: the
type and nature of the active agent being formulated; the subject to which the
agent-
containing composition is to be administered; the intended route of
administration of the
composition; and the therapeutic indication being targeted. Pharmaceutically
acceptable
carriers include both aqueous and non-aqueous liquid media, as well as a
variety of solid and
semi-solid dosage forms. Such carriers can include a number of different
ingredients and
additives in addition to the active agent, such additional ingredients being
included in the
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formulation for a variety of reasons, e.g., stabilization of the active agent,
binders, etc., well
known to those of ordinary skill in the art. Descriptions of suitable
pharmaceutically
acceptable carriers, and factors involved in their selection, are found in a
variety of readily
available sources such as, for example, Remington's Pharmaceutical Sciences,
18th Ed. (1990)
or Remington: The Science and Practice of Pharmacy, 22n1 Ed., Pharmaceutical
Press (2013)
or Introduction to Pharmaceutical Dosage Forms, 4th ed., Lea & Febiger,
Philadelphia (1985)
or Handbook of Pharmaceutical Excipients (Raymond C. Rowe et al., APhA
Publications, 4th
edition 2003) or Goodman & Gilman's The Pharmacological Basis of Therapeutics
(Joel G.
Hardman etal., eds., McGraw Hill Professional, 10th ed. 2001, or 13th ed.
2017) or
Pharmaceutical Dosage Forms and Drug Delivery Systems (Howard C. Ansel et al.,
eds.,
Lippincott Williams & Wilkins Publishers, 7th ed. 1999). Suitable
pharmaceutical
compositions may be formulated by means known in the art, or that become
available to
those skilled in the art, and their mode of administration and dose determined
by the skilled
practitioner. These protocols are routine proceedures and any modifications
are well within
the scope of one skilled in the art and from the teaching herein. In preferred
embodiments,
pharmaceutical compositions according to the invention are sterile
compositions.
A therapeutic compound disclosed herein may be formulated by itself in a
pharmaceutical
composition, or may be formulated together with one or more other therapeutic
compounds
disclosed herein in a single pharmaceutical composition. Depending upon the
particular
condition, or disease, to be treated, additional therapeutic agent(s) that are
normally
administered to treat that condition may also be present in the compositions
disclosed herein.
Some examples of materials which can serve as pharmaceutically acceptable
carriers include:
.. (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as
corn starch and potato
starch; (3) cellulose, and its analogs, such as sodium carboxymethyl
cellulose, ethyl cellulose
and cellulose acetate; (4) powdered tragacanth; (5) malt, (6) gelatin; (7)
talc.; (8) excipients,
such as cocoa butter and suppository waxes; (9) oils, such as peanut oil,
cotton seed oil,
safflower oil, sesame oil, olive oil, corn oil and soybean oil: (10) glycols,
such as propylene
glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene
glycol; (12) esters,
such as ethyl oleate and ethyl laurate; (13) agar, (14) buffering agents, such
as magnesium
hydroide and aluminum hydroxide; (15) alginic acid, (16) pyrogen-free water,
(17) isotonic
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saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer
solutions; and (21)
other non-toxic compatible substances employed in pharmaceutical formulations.
Buffers include, without limitation, acetate buffers, citrate buffers,
phosphate buffers, neutral
buffered saline, phosphate buffered saline and borate buffers. It is
understood that acids or
bases can be used to adjust the pH of a composition as needed.
Pharmaceutically acceptable
antioxidants include, without limitation, sodium metabisulfite, sodium
thiosulfate,
acetylcysteine, butylated hydroxyanisole and butylated hydroxytoluene, (1)
water soluble
antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate,
sodium
metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such
as ascorbyl
palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT),
lecithin,
propyl gallate, alpha-tocopherol, and the like; (3) metal chelating agents,
such as citric acid,
ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric
acid, and the like.
Useful preservatives include, without limitation, benzalkonium chloride,
chlorobutanol,
thimerosal, phenylmercuric acetate, phenylmercuric nitrate, a stabilized
oxychloro
composition and chelants, such as, e.g., DTPA or DTPA-bisamide, calcium DTPA,
and
CaNaDTPA-bisamide. Tonicity adjustors useful in a pharmaceutical composition
include,
without limitation, salts such as, e.g., sodium chloride, potassium chloride,
mannitol or
glycerin and other pharmaceutically acceptable tonicity adjustor.
In certain embodiments, the pharmaceutical preparation is non-pyrogenic i.e.
does not
substantially elevate the body temperature of a subject.
The amount of active ingredient that can be combined with a carrier material
to produce a
single dosage form will generally be that amount of the compound which
produces a
therapeutic effect. Generally, out of one hundred percent, this amount will
range from about 1
percent to about ninety-nine percent of active ingredient, preferably from
about 5 percent to
about 70 percent, most preferably from about 10 percent to about 30 percent.
Methods of preparing these formulations or compositions include the step of
bringing into
association a compound(s) of the present invention with the carrier and,
optionally, one or
more accessory ingredients. In general, the formulations are prepared by
uniformly and
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intimately bringing into association a compound of the present invention with
liquid carriers,
or finely divided solid carriers, or both, and then, if necessary, shaping the
product.
Administration
The compounds of Formula [X] may be administered by any means suitable for the
condition
to be treated e.g. parenteral administration e.g. by intramuscular,
intradermal, epicutaneous,
intraperitoneal, intravenous or intra-arterial (infusion or bolus, dose given
continuously or
intermittently), intracardiac, intracistemal, epidural, intrastemal,
intramedullary, intravesical
injection or infusion, via the cerebrospinal fluid, instillation, subcutaneous
injection/implant,
transdermal (e.g. by skin patch, controlled release patch), transmucosal
(e.g., nasal,
sublingual, vaginal, buccal, or rectal) or oral routes, all using dosage forms
well known to
those of ordinary skill in the pharmaceutical arts. For further example: oral,
parenteral,
enteral (e.g. oral, buccal, sublabial, lingual, sublingual), infusion,
injection, transdermal,
topical, paste applied to tongue, intravaginal (e.g. pessaries, tampons),
rectal (e.g. by
suppository, retention enema, enema, Murphy drip), transcutaneous,
intracutaneous,
subcutaneous, sublabial, subcuticular, intramuscular, intraarticular,
intracapsular,
subcapsular, subarachnoid, ocular/intraocular/ophthalmic (e.g. by eye drops
{optionally using
a measured dose eyedropper}, eye lotion), intraorbital, intravitreal,
retrobulbar, aural, ear
drops, intrathecal, intraventricular, intracardiac, intrasynovial,
intracerebral,
intracerebroventricular (optionally using a Ommaya reservoir), intraduodenal,
intradermal,
transdermal, intrapleural, intraspinal, intrastemal, intrathecal,
intralesional, intratumoral,
intracavemous (base of penis), extra-amniotic administration/infusion,
perivascular
administration, intravesicular, intraosseously (e.g. intraosseous infusion),
endotracheally,
transtracheal, intravaginal, nasally, by inhalation, insufflation (snorting),
by skin patch, by
dermal patch, by transdermal patch (e.g., without restriction, matrix type
patch, reservoir type
patch, monolithic drug-in-adhesive type patch, multilaminate drug-in-adhesive
type patch,
and the like, refer Ghosh TK, Pfister WR, Yum SI, Transdermal and Topical Drug
Delivery
Systems, Interpharm Press, Inc., one of ordinary skill in the art can
determine other patches
which can be employed in the present invention), by adhesive bandage,
transdermal spray,
transdermal implant, nanocell injection, patient controlled pump, peripherally
inserted central
catheter (PIC line), delivery by catheter, indwelling catheter,
gastric/duodenal feeding tube,
gastrostomy, enteral nutrition, balloon (drug-eluting balloon), stent (drug-
eluting stent),
vaginal ring, vaginal sponge, vaginal douche, intrauterine device, cervical
ring and the like,
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local injection into a body part(s) that is affected by a cancer(s). The
compound may be
delivered by
pill/tablet/capsule/pastille/time release technology/modified release
dosage/osmotic delivery
system/orally disintegrating tablet/film/lollipop/sublingual
drops/lozenges/effervescent
buccal tablet/chewing gum/smoking device/dry-powder
inhaler/vaporizer/nebulizer/metered-
dose inhaler/gas mask/nasal cannula/nasal spray/mucoadhesive
microdisc/pessary/suppository/electuary/emulsion/extended-release
syrup/effervescent
[powder/tablet]/hydrogel/molecular
=
encapsulation/powder/softgel/solution/suspension/syrup/syrup concentrate for
dilution and/or
addition of carbonated water/tincture/moutwash/toothpaste/ointment/oral
spray/nasal
spray/liniment/electrophoretic dermal delivery system/liposomes/transfersome
vesicles/lip
balm/shampoo/jet
injector/vapor/solid/decoction/ointment/cream/salve/oil/rinse/alcohol/spray/aer
osol/foam/past
e/lotion/tincture/shake
lotion/gel/implant/dressing/sponge/tape/drop/powder/patch/transdermal
patch/electroporation/iontophoresis/phonophoresis/sonophoresis, penetration
enhancers may
be incorporated e.g. see Finnin and Morgan, J. Pharm. Sci. 1999, 88, 955-958).
In other
embodiments, local administration may be accomplished by implanting a
sustained-release
device such as a pump or a micropump, or sustained-release implant, such as a
bead or gel
that contains the compound(s), e.g. anti-cancer agent, and slowly releases the
drug into the
desired area over time. Suitable devices for parenteral administration include
needle
(including microneedle) injectors, needle-free injectors and infusion
techniques. The
compound may be delivered orally, such as in the form of tablets(optionally
chewable)/caplets/capsules/pills/lozenges(including liquid
filled)/pastilles/electuary/paste/oral films/[buccal/mucoadhesive] patches
(each of which also
includes sustained release or timed release formulations), granules, pastilles
(e.g. using an
inert base, such as gelatin and glycerin, or sucrose and acacia), lozenges
(e.g. using a flavored
basis, usually sucrose and acacia or tragacanth), lyophilisates, dragees,
cachets, troches,
microgranules, pellets, soft-gels, powders, powders for reconstitution,
dispersions, tinctures,
or liquid formulations including syrups, liquids, solutions, elixirs,
suspensions (e.g., aqueous
or non-aqueous liquid suspensions, oil-in-water emulsions, or a water-in-oil
liquid
emulsions), emulsions, microemulsions, solutions, mouthwashes, elixirs and the
like, each
containing a predetermined amount of a compound(s) of the present invention as
an active
ingredient; liquid dosage forms suitable for parenteral administration to a
patient; eye drops
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or other ophthalmic preparations suitable for topical administration;
emulsions or magmas;
sublingually; bucally; transdermally; parenterally, such as by subcutaneous,
intravenous,
intramuscular or intrasternal injection or infusion (e.g., as sterile
injectable aqueous or non-
aqueous solutions or suspensions; sterile solids {e.g., crystalline or
amorphous solids} that
can be reconstituted to provide liquid dosage forms suitable for parenteral
administration);
nasally such as by inhalation spray (aerosol, nasal spray, inhaler, nebuliser
etc.); rectally such
as in the form of suppositories; or liposomally; each containing a
predetermined amount of a
compound(s) of the present invention as an active ingredient. A compound(s) of
the present
invention may also be administered as a bolus, electuary or paste. A
compound(s) of this
invention can be administered alone, but generally will be administered with a
pharmaceutical carrier selected on the basis of the chosen route of
administration and
standard pharmaceutical practice. Dosage unit formulations containing non-
toxic,
pharmaceutically acceptable vehicles or diluents may be administered. The
compounds may
be administered in a form suitable for immediate release or extended release.
Immediate
release or extended release may be achieved with suitable pharmaceutical
compositions or,
particularly in the case of extended release, with devices such as
subcutaneous implants or
osmotic pumps.
To illustrate, for oral administration in the form of a tablet or capsule, the
active drug
component can be combined with an oral, non-toxic, pharmaceutically
acceptable, inert
carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium
stearate,
dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like; for
oral administration
in liquid form, the oral drug components can be combined with any oral, non-
toxic,
pharmaceutically acceptable inert carrier such as ethanol, glycerol, water,
and the like.
Moreover, when desired or necessary, suitable binders, lubricants,
disintegrating agents, and
coloring agents can also be incorporated into the mixture. Suitable binders
include starch,
gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners,
natural and synthetic
gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose,
polyethylene
glycol, waxes, and the like. Lubricants used in these dosage forms include
sodium oleate,
sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium
chloride, and
the like. Disintegrators include, without limitation, starch, methyl
cellulose, agar, bentonite,
xanthan gum, and the like.
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In solid dosage forms of the invention for oral administration (capsules,
tablets, pills, dragees,
powders, granules and the like), the active ingredient is mixed with one or
more
pharmaceutically acceptable carriers, such as sodium citrate or dicalcium
phosphate, and/or
any of the following: (1) fillers or extenders, such as starches, lactose,
sucrose, glucose,
mannitol, and/or silicic acid; (2) binders, such as, for example,
carboxymethylcellulose,
alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3)
humectants, such as
glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate,
potato or tapioca
starch, alginic acid, certain silicates, and sodium carbonate; (5) solution
retarding agents,
such as paraffin; (6) absorption accelerators, such as quaternary ammonium
compounds; (7)
.. wetting agents, such as, for example, cetyl alcohol and glycerol
monostearate; (8) absorbents,
such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium
stearate, magnesium
stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures
thereof; and (10)
coloring agents. In the case of capsules, tablets and pills, the
pharmaceutical compositions
may also comprise buffering agents. Solid compositions of a similar type may
also be
employed as fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or
milk sugars, as well as high molecular weight polyethylene glycols and the
like.
A tablet may be made by compression or molding, optionally with one or more
accessory
ingredients. Compressed tablets may be prepared' using binder (for example,
gelatin or
hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative,
disintegrant (for
example, sodium starch glycolate or cross-linked sodium carboxymethyl
cellulose), surface-
active or dispersing agent. Molded tablets may be made by molding in a
suitable machine a
mixture of the powdered compound moistened with an inert liquid diluent.
The tablets, and other solid dosage forms of the pharmaceutical compositions
of the present
invention, such as dragees, capsules, pills and granules, may optionally be
scored or prepared
with coatings and shells, such as enteric coatings and other coatings well
known in the
pharmaceutical-formulating art. They may also be formulated so as to provide
slow or
controlled release of the active ingredient therein using, for example,
hydroxypropylmethyl
cellulose in varying proportions to provide the desired release profile, other
polymer
matrices, liposomes and/or microspheres. They may be sterilized by, for
example, filtration
through a bacteria-retaining filter, or by incorporating sterilizing agents in
the form of sterile
solid compositions which can be dissolved in sterile water, or some other
sterile injectable
medium immediately before use. These compositions may also optionally contain
opacifying
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agents and may be of a composition that they release the active ingredient(s)
only, or
preferentially, in a certain portion of the gastrointestinal tract,
optionally, in a delayed
manner. Examples of embedding compositions which can be used include polymeric
substances and waxes. The active ingredient can also be in micro-encapsulated
form, if
appropriate, with one or more of the above-described excipients.
Gelatin capsules may contain the active ingredient and powdered carriers, such
as lactose,
starch, cellulose derivatives, magnesium stearate, stearic acid, and the like.
Similar diluents
can be used to make compressed tablets. Both tablets and capsules can be
manufactured as
sustained release products to provide for continuous release of medication
over a period of
hours. Compressed tablets can be sugar coated or film coated to mask any
unpleasant taste
and protect the tablet from the atmosphere, or enteric coated for selective
disintegration in the
gastrointestinal tract.
Exemplary compositions for oral administration include suspensions which may
contain, for
example, microcrystal line cellulose for imparting bulk, alginic acid or
sodium alginate as a
suspending agent, methylcellulose as a viscosity enhancer, and sweeteners or
flavouring
agents such as those known in the art; and immediate release tablets which may
contain, for
example, microcrystalline cellulose, sugar(s), dicalcium phosphate, starch,
magnesium
stearate and/or lactose and/or other excipients, binders, extenders,
disintegrants, diluents,
granulating agents and lubricants such as those known in the art. The
inventive compounds
may be orally delivered by sublingual and/or buccal administration, e.g., with
molded,
compressed, or freeze-dried tablets. Exemplary compositions may include fast-
dissolving
diluents such as mannitol, lactose, sucrose, and/or cyclodextrins. Also
included in such
formulations may be high molecular weight excipients such as celluloses
(AVICEL0) or
polyethylene glycols (PEG); an excipient to aid mucosal adhesion such as
hydroxypropyl
cellulose (HPC), hydroxypropyl methyl cellulose (HPMC), sodium carboxymethyl
cellulose
(SCMC), and/or maleic anhydride copolymer (e.g., GANTREZ0); and agents to
control
release such as polyacrylic copolymer (e.g., CARBOPOL 9340). Lubricants,
glidants,
flavours, colouring agents and stabilizers may also be added for ease of
fabrication and use.
Liquid dosage forms for oral administration of the compounds of the invention
include
pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions,
syrups and
elixirs. In addition to the active ingredient, the liquid dosage forms may
contain inert diluents
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commonly used in the art, such as, for example, water or other solvents,
solubilizing agents
and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate,
ethyl acetate,
benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils
(in particular,
cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol,
tetrahydrofuryl
alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures
thereof.
Liquids for oral administration may be in the form of suspensions, solutions,
emulsions, or
syrups, or may be lyophilized or presented as a dry product for reconstitution
with water or
other suitable vehicle before use. Such liquid compositions may optionally
contain:
pharmaceutically-acceptable excipients such as preservatives, buffers,
propellants,
suspending agents (for example, sorbitol, methyl cellulose, sodium alginate,
gelatin,
hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel and the
like); non-
aqueous vehicles, e.g., oil (for example, almond oil or fractionated coconut
oil), propylene
glycol, ethyl alcohol, or water; preservatives (for example, methyl or propyl
p-
hydroxybenzoate or sorbic acid); wetting agents such as lecithin; and, if
desired, flavoring or
coloring agents. Indeed, liquid dosage forms for oral administration can
contain coloring and
flavoring to increase patient acceptance.
Suspensions, in addition to the active compounds, may contain suspending
agents as, for
example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and
sorbitan esters,
microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and
tragacanth,
and mixtures thereof.
Oily Suspensions may be formulated by suspending a therapeutic compound
disclosed herein
in admixture with (a) Vegetable oils, such as, e.g., almond oil, arachis oil,
avocado oil, canola
oil, castor oil, coconut oil, corn oil, cottonseed oil, grape seed oil,
hazelnut oil, hemp oil,
linseed oil, olive oil, palm oil, peanut oil, rapeseed oil, rice bran oil,
safflower oil, sesame oil,
soybean oil, soya oil, sunflower oil, walnut oil, wheat germ oil, or a
combination thereof, (b)
a saturated fatty acid, an unsaturated fatty acid, or a combination thereof.
Such as, e.g.,
palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, or a
combination thereof,
(c) mineral oil such as, e.g., liquid paraffin, (d) Surfactants or detergents.
The oily
suspensions may contain a thickening agent, for example beeswax, hard paraffin
or cetyl
alcohol. Sweetening agents and flavoring agents may be added to provide a
palatable oral
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preparation. These compositions may be preserved by the addition of an
antioxidant such as
ascorbic acid.
A therapeutic compound disclosed herein may be in the form of oil-in-water
emulsions. The
oily phase may be a vegetable oil as disclosed herein or a mineral oil as
disclosed herein or
mixtures thereof. Suitable emulsifying agents may be naturally occurring gums,
such as, e.g.,
gum acacia or gum tragacanth, naturally occurring phosphatides, for example
soya bean,
lecithin, and esters or partial esters derived from fatty acids and hexitol
anhydrides, for
example sorbitan monooleate and condensation products of the said partial
esters with
ethylene oxide, for example polyoxyethylene sorbitan monooleate.
Exemplary compositions for nasal aerosol or inhalation administration include
solutions
which may contain, for example, benzyl alcohol or other suitable
preservatives, absorption
promoters to enhance absorption and/or bioavailability, and/or other
solubilizing or
dispersing agents such as those known in the art.
Pharmaceutical compositions of this invention suitable for parenteral
administration comprise
one or more compounds of the invention in combination with one or more
pharmaceutically
acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions,
suspensions or
emulsions, or sterile powders which may be reconstituted into sterile
injectable solutions or
dispersions just prior to use, which may contain antioxidants, buffers,
bacteriostats, solutes
which render the formulation isotonic with the blood of the intended
recipient, or suspending
or thickening agents.
Examples of suitable aqueous and nonaqueous carriers which may be employed in
the
pharmaceutical compositions of the invention include water, ethanol, polyols
(such as
glycerol, propylene glycol, polyethylene glycol, and the like), and suitable
mixtures thereof,
vegetable oils, such as olive oil, and injectable organic esters, such as
ethyl oleate. Proper
fluidity can be maintained, for example, by the use of coating materials, such
as lecithin, by
the maintenance of the required particle size in the case of dispersions, and
by the use of
surfactants.
These compositions may also contain adjuvants such as preservatives, wetting
agents,
emulsifying agents and dispersing agents. Inhibition of the action of
microorganisms may be
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ensured by the inclusion of various antibacterial and antifungal agents, for
example, paraben,
chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to
include isotonic
agents, such as sugars, sodium chloride, and the like into the compositions.
In addition,
prolonged absorption of the injectable pharmaceutical form may be brought
about by the
inclusion of agents which delay absorption, such as aluminum monostearate and
gelatin.
In some cases, in order to prolong the effect of a drug, it is desirable to
slow the absorption of
the drug from subcutaneous or intramuscular injection. This may be
accomplished by the use
of a liquid suspension of crystalline or amorphous material having poor water
solubility. The
rate of absorption of the drug then depends upon its rate of dissolution
which, in turn, may
depend upon crystal size and crystalline form. Alternatively, delayed
absorption of a
parenterally administered drug form is accomplished by dissolving or
suspending the drug in
an oil vehicle.
.. Injectable depot forms are made by forming microencapsule matrices of the
subject
compounds in biodegradable polymers such as polylactide-polyglycolide.
Depending on the
ratio of drug to polymer, and the nature of the particular polymer employed,
the rate of drug
release can be controlled. Examples of other biodegradable polymers include
poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also
prepared by
entrapping the drug in liposomes or microemulsions which are compatible with
body tissue.
Exemplary compositions for parenteral administration include injectable
solutions or
suspensions, preferably sterile and preferably buffered to an appropriate pH
and isotonicity,
and/or which may contain, for example, suitable non-toxic, parenterally
acceptable diluents,
oils or solvents, such as mannitol, 1,3-butanediol, water, Ringer's solution,
an isotonic
sodium chloride solution, or other suitable dispersing or wetting and
suspending agents,
including synthetic mono- or diglycerides, and fatty acids, including oleic
acid, ethyl alcohol,
polyethylene glycol, polypropylene glycol, corn oil, cottonseed oil, peanut
oil, sesame oil,
ethyl oleate, isopropyl myristate, and benzyl benzoate. In general, water, a
suitable oil, saline,
aqueous dextrose (glucose), and related sugar solutions and glycols such as
propylene glycol
or polyethylene glycols are suitable carriers for parenteral solutions.
Solutions for parenteral
administration preferably contain a water soluble salt of the active
ingredient, suitable
stabilizing agents, and if necessary, buffer substances. Antioxidizing agents
such as sodium
bisulfite, sodium sulfite, or ascorbic acid, either alone or combined, are
suitable stabilizing
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agents. Also used are citric acid and its salts and sodium EDTA. In addition,
parenteral
solutions can contain preservatives, such as benzalkonium chloride, methyl- or
propyl-
paraben, and chlorobutanol. Such forms may be presented in unit-dose form such
as
ampoules or disposable injection devices, in multi-dose forms such as vials
from which the
appropriate dose may be withdrawn, or in a solid form or pre-concentrate that
can be used to
prepare an injectable formulation. Compounds that increase the solubility of
one or more of
the active ingredients disclosed herein can also be incorporated into the
parenteral dosage
forms provided herein. For example, cyclodextrin and/or its derivatives can be
used to
increase the solubility of a compound provided herein.
Sterile compositions of compound(s) of this invention are contemplated by the
invention,
indeed preferred, including compositions that are in accord with national and
local
regulations governing such compositions.
Formulations of the pharmaceutical compositions of the invention for rectal or
vaginal
administration may be presented as a suppository, which may be prepared by
mixing one or
more compounds of the invention with one or more suitable nonirritating
excipients or
carriers comprising, for example, cocoa butter, polyethylene glycol, a
suppository wax or a
salicylate, and which is solid at room temperature, but liquid at body
temperature and,
therefore, will melt in the rectum or vaginal cavity and release the active
ingredient.
Exemplary compositions for rectal/vaginal administration include suppositories
which may
contain, for example, suitable non-irritating excipients, such as cocoa
butter, synthetic
glyceride esters or polyethylene glycols, which are solid at ordinary
temperatures but liquefy
and/or dissolve in the rectal/vaginal cavity to release the drug. Formulations
of the present
invention which are suitable for vaginal administration also include
pessaries, tampons,
creams, gels, pastes, foams or spray formulations containing such carriers as
are known in the
art to be appropriate.
Dosage forms for the topical or transdermal administration of a compound of
this invention
include powders, sprays, ointments, pastes, creams, lotions, gels, solutions,
patches and
inhalants. The active compound may be mixed under sterile conditions with a
pharmaceutically acceptable carrier, and with any preservatives, buffers, or
propellants which
may be required.
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The ointments, pastes, creams and gels may contain, in addition to an active
compound of
this invention, excipients, such as animal and vegetable fats, oils, waxes,
paraffins, starch,
tragacanth, cellulose analogs, polyethylene glycols, silicones, bentonites,
silicic acid, talc and
zinc oxide, or mixtures thereof.
Powders and sprays can contain, in addition to a compound of this invention,
excipients such
as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and
polyamide powder, or
mixtures of these substances. Sprays can additionally contain customary
propellants, such as
chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as
butane and
propane.
Ophthalmic formulations, eye ointments, powders, solutions, drops and the
like, are also
contemplated as being within the scope of this invention.
Transdermal patches have the added advantage of providing controlled delivery
of a
compound of the present invention to the body. Such dosage forms can be made
by
dissolving or dispersing the composition in the proper medium. Absorption
enhancers can
also be used to increase the flux of the composition across the skin. The rate
of such flux can
be controlled by either providing a rate-controlling membrane or dispersing
the compound in
a polymer matrix or gel.
In some embodiments, a compound or pharmaceutical formulation in accordance
may be
administered by means of a medical device or appliance such as an implant,
graft, prosthesis,
stent etc. Implants may be devised which are intended to contain and release
such compounds
or compositions. An example would be an implant made of a polymeric material
adapted to
release the compound over a period of time.
The compounds of the present invention can also be administered in the form of
liposome
delivery systems, such as small unilamellar vesicles, large unilamellar
vesicles, and
multilamellar vesicles. Liposomes can be formed from a variety of
phospholipids, such as
cholesterol, stearylamine, or phosphatidylcholines. Any non-toxic,
physiologically acceptable
and metabolizable lipid capable of forming liposomes can be used. The
compounds described
herein, when in liposome form can contain, in addition to the compounds
described herein,
stabilizers, preservatives, excipients, and the like. Methods to form
liposomes are known in
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the art. (See, e.g., Prescott, Ed., Methods in Cell Biology, Volume XIV,
Academic Press,
New York, N.Y., (1976), P33 et seq.).
Compounds of the present invention may also be coupled with soluble polymers
as targetable
drug carriers. Such polymers can include polyvinylpyrrolidone, pyran
copolymer,
polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol, or
polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore,
the
compounds of the present invention may be coupled to a class of biodegradable
polymers
useful in achieving controlled release of a drug, for example, polylactic
acid, polyglycolic
acid, copolymers of polylactic and polyglycolic acid, polyepsilon
caprolactone, polyhydroxy
butyric acid, polyorthoesters, polyacetals, polydihydropyrans,
polycyanoacylates, and
crosslinked or amphipathic block copolymers of hydrogels.
The composition, shape, and type of dosage forms provided herein may vary
depending on
their use. For example, a dosage form used in the acute treatment of a disease
may contain
larger amounts of one or more of the active ingredients than a dosage form
used in the
chronic treatment of the same disease. Similarly, a parenteral dosage form may
contain
smaller amounts of one or more of the active ingredients than an oral dosage
form used to
treat the same disease. See, e.g., Remington's Pharmaceutical Sciences, 18th
ed., Mack
Publishing, Easton Pa. (1990).
Whether a particular excipient is suitable for incorporation into a
pharmaceutical composition
or dosage form provided herein depends on a variety of factors, including, but
not limited to,
the route of administration. For example, oral dosage forms such as tablets
may contain
excipients not suited for use in parenteral dosage forms. The suitability of a
particular
excipient may also depend on the specific active ingredients in the dosage
form. For example,
the decomposition of some active ingredients may be accelerated by some
excipients such as
lactose, or when exposed to water. Active ingredients that comprise primary or
secondary
amines are particularly susceptible to such accelerated decomposition.
Consequently,
encompassed herein are pharmaceutical compositions and dosage forms that
contain little, if
any, lactose. As used herein, the term "lactose-free" means that the amount of
lactose present,
if any, is insufficient to substantially increase the degradation rate of an
active ingredient.
Lactose-free compositions provided herein can comprise excipients that are
listed, for
example, in the U.S. Pharmacopeia (USP) 25-NF20 (2002). In certain
embodiments, lactose-
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free compositions comprise active ingredients, a binder/filler, and a
lubricant in
pharmaceutically compatible and pharmaceutically acceptable amounts. In
certain
embodiments, lactose-free dosage forms comprise active ingredients,
microcrystalline
cellulose, pre-gelatinized starch, and magnesium stearate.
Further encompassed herein are anhydrous pharmaceutical compositions and
dosage
forms comprising active ingredients, since water can facilitate the
degradation of some
compounds. For example, the addition of water (e.g., 5%) is widely accepted in
the
pharmaceutical arts as a means of simulating long-term storage in order to
determine
characteristics such as shelf-life or the stability of formulations over time.
See, e.g., Jens T.
Carstensen, Drug Stability: Principles & Practice, 2d. Ed., Marcel Dekker, NY,
N.Y., 1995,
pp. 379-80. In effect, water and heat accelerate the decomposition of some
compounds. Thus,
the effect of water on a formulation can be of great significance since
moisture and/or
humidity are commonly encountered during manufacture, handling, packaging,
storage,
shipment, and use of formulations.
Anhydrous pharmaceutical compositions and dosage forms provided herein can be
prepared using anhydrous or low moisture containing ingredients and low
moisture or low
humidity conditions. Pharmaceutical compositions and dosage forms that
comprise lactose
and at least one active ingredient that comprises a primary or secondary amine
are preferably
anhydrous if substantial contact with moisture and/or humidity during
manufacturing,
packaging, and/or storage is expected.
An anhydrous pharmaceutical composition should be prepared and stored such
that its
anhydrous nature is maintained. Accordingly, in certain embodiments, provided
herein are
anhydrous compositions packaged using materials to prevent exposure to water
such that they
can be included in suitable formulary kits. Examples of suitable packaging
include, but are
not limited to, hermetically sealed foils, plastics, unit dose containers
(e.g., vials), blister
packs, and strip packs.
Encompassed herein are pharmaceutical compositions and dosage forms that
comprise
one or more compounds that reduce the rate by which an active ingredient will
decompose.
Such compounds, which are referred to herein as "stabilizers," include, but
are not limited to,
antioxidants such as ascorbic acid, pH buffers, or salt buffers.
Examples of pharmaceutically-acceptable antioxidants include, but are not
limited to: (1)
water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride,
sodium bisulfate,
sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble
antioxidants, such as
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ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxy toluene
(BHT),
lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal
chelating agents, such as
citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid,
phosphoric acid,
and the like.
The pH of a pharmaceutical composition or dosage form may also be adjusted to
improve
delivery of one or more active ingredients. Similarly, the polarity of a
solvent carrier, its ionic
strength, or tonicity can be adjusted to improve delivery. Compounds such as
stearates can
also be added to pharmaceutical compositions or dosage forms to advantageously
alter the
hydrophilicity or lipophilicity of one or more active ingredients so as to
improve delivery. In
this regard, stearates can serve as a lipid vehicle for the formulation, as an
emulsifying agent
or surfactant, and as a delivery-enhancing or penetration-enhancing agent.
Different salts,
hydrates or solvates of the active ingredients can be used to further adjust
the properties of
the resulting composition.
Consumable oral films for human or veterinary use are typically pliable water-
soluble or
water-swellable thin film dosage forms which may be rapidly dissolving or
mucoadhesive
and typically comprise a compound(s) of Formula [X], a film-forming polymer, a
binder, a
solvent, a humectant, a plasticizer, a stabilizer or emulsifier, a viscosity-
modifying agent and
a solvent. Some components of the formulation may perform more than one
function.
Alternatively, the compound(s) of Formula [X] may be in the form of
multiparticulate beads.
The film-forming polymer may be selected from natural polysaccharides,
proteins, or
synthetic hydrocolloids and is typically present in the range 0.01 to 99
weight %, more
typically in the range 30 to 80 weight %. Films in accordance with the
invention are typically
prepared by evaporative drying of thin aqueous films coated onto a peelable
backing support
or paper. This may be done in a drying oven or tunnel, typically a combined
coater dryer, or
by freeze-drying or vacuuming.
The compounds of the invention (including pharmaceutically acceptable salts
thereof) can
also be administered intranasally or by inhalation, typically in the form of a
dry powder
(either alone; as a mixture, for example, in a dry blend with lactose; or as a
mixed component
particle, for example, mixed with phospholipids, such as phosphatidylcholine)
from a dry
powder inhaler, as an aerosol spray from a pressurized container, pump, spray,
atomizer (for
example an atomizer using electrohydrodynamics to produce a fine mist), or
nebulizer, with
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or without the use of a suitable propellant, such as 1,1,1,2-tetrafluoroethane
or 1,1,1,2,3,3,3-
heptafluoropropane, or as nasal drops. For intranasal use, the powder may
comprise a
bioadhesive agent, for example, chitosan or cyclodextrin.
The pressurized container, pump, spray, atomizer, or nebulizer contains a
solution or
suspension of the compound(s) of the invention comprising, for example,
ethanol, aqueous
ethanol, or a suitable alternative agent for dispersing, solubilizing, or
extending release of the
active, a propellant(s) as solvent and an optional surfactant, such as
sorbitan trioleate, oleic
acid, or an oligolactic acid.
Prior to use in a dry powder or suspension formulation, the drug product is
micronized to
a size suitable for delivery by inhalation (typically less than 5 microns).
This may be
achieved by any appropriate comminuting method, such as spiral jet milling,
fluid bed jet
milling, supercritical fluid processing to form nanoparticles, high pressure
homogenization,
or spray drying.
In inhaled dosage forms, the therapeutic compound(s) may be prepared for
delivery as an
aerosol in a liquid propellant for use in a pressurised (PD!) or other metered
dose inhaler
(MDI). Propellants suitable for use in a PD! or MDI include, without
limitation, CFC-12,
HFA-134a, HFA-227, HCFC-22 (difluorochloromethane), HFA-152 (difluoroethane
and
isobutane). A therapeutic compound may also be delivered using a nebulisers or
other aerosol
delivery system. A therapeutic compound may be prepared for delivery as a dry
powder for
use in a dry powder inhaler (DPI). A dry powder for use in the inhalers will
usually have a
mass median aerodynamic diameter of less than 30 pm, preferably less than 20
pm and more
preferably less than 10 pm. Microparticles having aerodynamic diameters in the
range of
about 5 pm to about 0.5 pm will generally be deposited in the respiratory
bronchioles,
whereas smaller particles, having aerodynamic diameters in the range of about
2 pm to about
0.05 pm, are likely to be deposited in the alveoli. A DPI may be a passive
delivery
mechanism, which relies on the individual's inspiration to introduce the
particles into the
lungs, or an active delivery mechanism, requiring a mechanism for delivering
the powder to
the individual. In inhalatory formulations, a therapeutically effective amount
of a therapeutic
compound disclosed herein for an inhaled formulation may be between about
0.0001% (w/v)
to about 60% (w/v), about 0.001% (w/v) to about 40.0% (w/v), or about 0.01%
(w/v) to about
20.0% (w/v). In inhalatory formulations, a therapeutically effective amount of
a therapeutic
compound disclosed herein for an inhaled formulation may also be between about
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0.0001% (w/w) to about 60% (w/w), about 0.001% (w/w) to about 40.0% (w/w), or
about
0.01% (w/w) to about 20.0% (w/w).
Tablets
Tablets may be uncoated or they may be coated by known techniques to delay
disintegration
and absorption in the gastrointestinal tract and thereby provide a sustained
action over a
longer period. For example, a time delay material such as glyceryl
monostearate or glyceryl
distearate may be employed.
Because of their ease of administration, tablets and capsules represent the
most advantageous
oral dosage unit forms, in which case solid excipients are employed. If
desired, tablets can be
coated by standard aqueous or nonaqueous techniques. Such dosage forms may be
prepared
by some known methods of pharmacy. In certain embodiments, pharmaceutical
compositions
and dosage forms are prepared by uniformly and intimately admixing the active
ingredients
with liquid carriers, finely divided solid carriers, or both, and then shaping
the product into
the desired presentation if necessary.
In certain embodiments, a tablet is prepared by compression or molding. In
certain
embodiments, compressed tablets are be prepared by compressing in a suitable
machine the
active ingredients in a free-flowing form, e.g., powder or granules,
optionally mixed with an
excipient. In certain embodiments, molded tablets are made by molding in a
suitable machine
a mixture of a powdered compound moistened with an inert liquid diluent.
Examples of excipients that can be used in oral dosage forms provided herein
include, but
are not limited to, binders, fillers, disintegrants, and lubricants. Binders
suitable for use in
pharmaceutical compositions and dosage forms provided herein include, but are
not limited
to, corn starch, potato starch, or other starches, gelatin, natural and
synthetic gums such as
acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth,
guar gum,
cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate,
carboxymethyl cellulose
calcium, sodium carboxymethyl cellulose), polyvinyl pyrrolidone, methyl
cellulose, pre-
gelatinized starch, hydroxypropyl methyl cellulose, (e.g., Nos. 2208, 2906,
2910),
microcrystalline cellulose, and mixtures thereof.
Suitable forms of microcrystalline cellulose include, but are not limited to,
AVICEL-PH-
101, AVICEL-PH-103 AVICEL RC-581, AVICEL-PH-105 (FMC Corporation, American
Viscose Division, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. An
specific binder
is a mixture of microcrystalline cellulose and sodium carboxymethyl cellulose
(e.g., AVICEL
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RC-581). Suitable anhydrous or low moisture excipients or additives include
AVICEL-PH-
103Tm and Starch 1500 LM.
Examples of fillers suitable for use in the pharmaceutical compositions and
dosage forms
provided herein include, but are not limited to, talc, calcium carbonate
(e.g., granules or
powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin,
mannitol, silicic
acid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof. In
certain embodiments,
the binder or filler in pharmaceutical compositions provided herein is present
in from about
50 to about 99 weight percent of the pharmaceutical composition or dosage
form.
Disintegrants are used in the compositions provided herein to provide tablets
the ability to
disintegrate when exposed to an aqueous environment. Tablets that contain too
much
disintegrant may disintegrate in storage, while those that contain too little
may not
disintegrate at a desired rate or under the desired conditions. Thus, a
sufficient amount of
disintegrant that is neither too much nor too little to detrimentally alter
the release of the
active ingredients should be used to form solid oral dosage forms provided
herein. The
amount of disintegrant used varies based upon the type of formulation. In
certain
embodiments, the pharmaceutical compositions provided herein comprise from
about 0.5 to
about 15 weight percent or from about 1 to about 5 weight percent of
disintegrant.
Disintegrants that are suitable for use in pharmaceutical compositions and
dosage forms
provided herein include, but are not limited to, agar-agar, alginic acid,
calcium carbonate,
microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin
potassium,
sodium starch glycolate, potato or tapioca starch, other starches, pre-
gelatinized starch, other
starches, clays, other algins, other celluloses, gums, and mixtures thereof.
Lubricants that are suitable for use in pharmaceutical compositions and dosage
forms
provided herein include, but are not limited to, calcium stearate, magnesium
stearate, mineral
oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol,
other glycols, stearic
acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut
oil, cottonseed oil,
sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc
stearate, ethyl oleate, ethyl
laureate, agar, and mixtures thereof. Additional lubricants include, but are
not limited to, a
syloid silica gel (AEROSIL200, W.R. Grace Co., Baltimore, Md.), a coagulated
aerosol of
synthetic silica (Degussa Co. of Plano, Tex.), CAB-O-SIL (a pyrogenic silicon
dioxide,
Cabot Co. of Boston, Mass.), and mixtures thereof. In certain embodiments, if
used at all,
lubricants are used in an amount of less than about 1 weight percent of the
pharmaceutical
compositions or dosage forms into which they are incorporated.
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In certain embodiments, provided herein is a solid oral dosage form,
comprising a
compound(s) of Formula [X], or an enantiomer or a mixture of enantiomers
thereof, or a
pharmaceutically acceptable salt, solvate, hydrate, prodrug, co-crystal,
clathrate, or
polymorph thereof; and one or more excipients selected from anhydrous lactose,
microcrystalline cellulose, polyvinylpyrrolidone, stearic acid, colloidal
anhydrous silica, and
gelatin.
If required, the solubility and bioavailability of the compounds of the
present invention in
pharmaceutical compositions may be enhanced by methods well-known in the art.
One
method includes the use of lipid excipients in the formulation. See "Oral
Lipid-Based
Formulations: Enhancing the Bioavailability of Poorly Water-Soluble Drugs
(Drugs and the
Pharmaceutical Sciences)," David J. Hauss, ed. Informa Healthcare, 2007; and
"Role of Lipid
Excipients in Modifying Oral and Parenteral Drug Delivery: Basic Principles
and Biological
Examples," Kishor M. Wasan, ed. Wiley-Interscience, 2006. Another known method
of
enhancing bioavailability is the use of an amorphous form of a compound of
this invention
optionally formulated with a poloxamer, such as LUTROLTm and PLURONICTM (BASF
Corporation), or block copolymers of ethylene oxide and propylene oxide.
Controlled release
Administration of compounds in controlled release formulations is useful where
the
compound of the present invention has
(i) a narrow "therapeutic index" e.g. the difference between the plasma
concentration leading
to harmful side effects or toxic reactions and the plasma concentration
leading to a
therapeutic effect is small; generally, the therapeutic index (TI) is defined
as the ratio of
median lethal dose (LD50), or median toxic dose (TD50), to median effective
dose (ED50);
(ii) a narrow absorption window in the gastro-intenstinal tract; or
(iii) a short biological half-life, such that frequent dosing during a day is
required in order to
sustain the plasma concentration at a therapeutic level.
Many strategies can be pursued to obtain controlled release in which the rate
of release
outweighs the rate of metabolism of the therapeutic compound. For example,
controlled
release can be obtained by the appropriate selection of formulation parameters
and
ingredients, including e.g. appropriate controlled release compositions and
coatings.
Examples include single or multiple unit tablet or capsule compositions, oil
solutions,
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suspensions, emulsions, microcapsules, microspheres, nanoparticle
formulations, patches,
and liposomes.
In certain embodiments, the active ingredients provided herein are
administered by controlled
.. release means or by delivery devices. Examples include, but are not limited
to, those
described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123;
4,008,719,
5,674,533, 5,059,595, 5,591,767, 5,120,548, 5,073,543, 5,639,476, 5,354,556,
and 5,733,566,
each of which is incorporated herein by reference in its entirety. In certain
embodiments, such
dosage forms are be used to provide slow or controlled-release of one or more
active
ingredients using, for example, hydropropylmethyl cellulose, other polymer
matrices, gels,
permeable membranes, osmotic systems, multilayer coatings, microparticles,
liposomes,
microspheres, or a combination thereof to provide the desired release profile
in varying
proportions. Encompassed herein are single unit dosage forms suitable for oral
administration, including, but not limited to, tablets, capsules, gelcaps, and
caplets that are
adapted for controlled-release.
All controlled-release pharmaceutical products have a common goal of improving
drug
therapy over that achieved by their non-controlled counterparts. Ideally, the
use of an
optimally designed controlled-release preparation in medical treatment is
characterized by a
minimum of drug substance being employed to cure or control the condition in a
minimum
amount of time. Advantages of controlled-release formulations include extended
activity of
the drug, reduced dosage frequency, and increased patient compliance. In
addition,
controlled-release formulations can be used to affect the time of onset of
action or other
characteristics, such as blood levels of the drug, and can thus affect the
occurrence of side
(e.g., adverse) effects.
Most controlled-release formulations are designed to initially release an
amount of drug
(active ingredient) that promptly produces the desired therapeutic effect, and
gradually and
continually release of other amounts of drug to maintain this level of
therapeutic or
prophylactic effect over an extended period of time. In order to maintain this
constant level of
drug in the body, the drug must be released from the dosage form at a rate
that will replace
the amount of drug being metabolized and excreted from the body. Controlled-
release of an
active ingredient can be stimulated by various conditions including, but not
limited to, pH,
temperature, enzymes, water, or other physiological conditions or compounds.
Controlled release II
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Compound(s) herein may also be formulated so as to provide slow or controlled
release of
this compound(s) using, for example, hydroxypropylmethyl cellulose in varying
proportions
to provide the desired release profile, other polymer matrices, liposomes,
microemulsions
and/or microspheres. They may be formulated for rapid release, e.g., freeze-
dried. These
compositions may also optionally contain opacifying agents and may be of a
composition that
they release the active ingredient(s) only, or preferentially, in a certain
portion of the
gastrointestinal tract, optionally, in a delayed manner. Examples of embedding
compositions
which can be used include aluminum monostearate, gelatin, polymeric substances
and waxes.
The active ingredient can also be in micro-encapsulated form, if appropriate,
with one or
more of the excipients herein or known to those of the art.
Modified release formulations include delayed-, sustained-, pulsed-,
controlled-, targeted and
programmed release. Suitable modified release formulations for the purposes of
the invention
are described in U.S. Pat. No. 6,106,864. Details of other suitable release
technologies such
as high energy dispersions and osmotic and coated particles are to be found in
Verma et al.,
Pharmaceutical Technology On-line, 25(2), 1-14 (2001). The use of chewing gum
to achieve
controlled release is described in WO 00/35298. Compounds of the invention may
be
formulated as a suspension or as a solid, semi-solid, or thixotropic liquid
for administration as
an implanted depot providing modified release of the active compound. Examples
of such
formulations include drug-coated stents and semi-solids and suspensions
comprising drug-
loaded poly(DL-lactic-coglycolic acid) (PLGA) microspheres.
In some cases, in order to prolong the effect of a drug, it is desirable to
slow the absorption of
the drug. This may be accomplished by the use of a liquid suspension of
crystalline or
amorphous material having poor water solubility. The rate of absorption of the
drug then
depends upon its rate of dissolution which, in turn, may depend upon crystal
size and
crystalline form. Alternatively, delayed absorption of an administered drug
form is
accomplished by dissolving or suspending the drug in an oil vehicle or by
forming and using
microencapsuled matrices of the subject compounds in biodegradable polymers
such as
polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the
nature of the
particular polymer employed, the rate of drug release can be controlled.
Examples of other
biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot
injectable
formulations are also prepared by entrapping the drug in liposomes or
microemulsions which
are compatible with body tissue.
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A therapeutic compound(s) disclosed herein, or a composition comprising such a
therapeutic
compound(s), may be incorporated into a drug delivery platform in order to
achieve a
controlled release profile over time. Such a drug delivery platform comprises
a therapeutic
compound(s) disclosed herein dispersed within a polymer matrix, typically a
biodegradable,
bioerodible, and/or bioresorbable polymer matrix. As used herein, the term
"polymer" refers
to synthetic homo- or copolymers, naturally occurring homo- or copolymers, as
well as
synthetic modifications or derivatives thereof having a linear, branched or
star structure.
Copolymers can be arranged in any form, such as, e.g., random, block,
segmented, tapered
blocks, graft, or triblock. Polymers are generally condensation polymers.
Polymers can be
further modified to enhance their mechanical or degradation properties by
introducing cross-
linking agents or changing the hydrophobicity of the side residues. If
crosslinked, polymers
are usually less than 5% crosslinked, usually less than 1% crosslinked.
Suitable polymers include, without limitation, alginates, aliphatic
polyesters, polyalkylene
oxalates, polyamides, polyamidoesters, polyanhydrides, polycarbonates,
polyesters,
polyethylene glycol, polyhydroxyaliphatic carboxylic acids, polyorthoesters,
polyoxaesters,
polypeptides, polyphosphazenes, polysaccharides, and polyurethanes. The
polymer usually
comprises at least about 10% (w/w), at least about 20% (w/w), at least about
30% (w/w), at
.. least about 40% (w/w), at least about 50% (w/w), at least about 60% (w/w),
at least about
70% (w/w), at least about 80% (w/w), or at least about 90% (w/w) of the drug
delivery
platform. Examples of biodegradable, bioerodible, and/or bioresorbable
polymers and
methods useful to make a drug delivery platform are described in, e.g., Drost,
et. al.,
Controlled Release Formulation, U.S. Pat. No. 4,756,911; Smith, et. al.,
Sustained Release
Drug Delivery Devices, U.S. Pat. No. 5,378.475; Wong and Kochinke. Formulation
for
Controlled Release of Drugs by Combining Hyrophilic and Hydrophobic Agents,
U.S. Pat.
No. 7,048,946; Hughes, et. al., Compositions and Methods for Localized Therapy
of the Eye,
U.S. Patent Publication 2005/0181017: Hughes, Hypotensive Lipid-Containing
Biodegradable Intraocular Implants and Related Methods, U.S. Patent
Publication
2005/0244464; Altman, et al., Silk Fibroin Hydrogels and Uses Thereof, U.S.
Patent
Publication 2011/0008437; each of which is incorporated by reference in its
entirety.
In aspects of this embodiment, a polymer composing the matrix is a polypeptide
such as, e.g.,
silk fibroin, keratin, or collagen. In other aspects of this embodiment, a
polymer composing
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the matrix is a polysaccharide such as, e.g., cellulose, agarose, elastin,
chitosan, chitin, or a
glycosaminoglycan like chondroitin sulfate, dermatan sulfate, keratan sulfate,
or hyaluronic
acid. In yet other aspects of this embodiment, a polymer composing the matrix
is a polyester
such as, e.g., D-lactic acid, L-lactic acid, racemic lactic acid, glycolic
acid, caprolactone, and
combinations thereof.
One of ordinary skill in the art appreciates that the selection of a suitable
polymer for forming
a suitable disclosed drug delivery platform depends on several factors. The
more relevant
factors in the selection of the appropriate polymer(s), include, without
limitation,
compatibility of polymer with drug, desired release kinetics of drug, desired
biodegradation
kinetics of platform at implantation site, desired bioerodible kinetics of
platform at
implantation site, desired bioresorbable kinetics of platform at implantation
site, in vivo
mechanical performance of platform, processing temperatures, biocompatibility
of platform,
and patient tolerance. Other relevant factors that, to some extent, dictate
the in vitro and in
vivo behavior of the polymer include the chemical composition, spatial
distribution of the
constituents, the molecular weight of the polymer and the degree of
crystallinity.
A drug delivery platform includes both a sustained release drug delivery
platform and an
extended release drug delivery platform. "Sustained release" can refer to the
release of a
therapeutic compound disclosed herein over a period of about seven days or
more.
"Extended release" can refer to the release of a therapeutic compound
disclosed herein over a
period of time of less than about seven days. In aspects of this embodiment, a
drug delivery
platform (e.g. sustained/extended release drug delivery platform) releases a
therapeutic
compound(s) disclosed herein with substantially zero or first order release
kinetics over an
illustrative (not restrictive) period of, e.g., about 1 day after
administration, about 2 days after
administration, about 3 days after administration, about 4 days after
administration, about 5
days after administration, about 6 days after administration, about 7 days
after administration,
about 15 days after administration, about 30 days after administration, about
45 days after
administration, about 60 days after administration, about 75 days after
administration, or
about 90 days after administration. In other aspects of this embodiment, a
drug delivery
platform (e.g. sustained/extended release drug delivery platform) releases a
therapeutic
compound(s) disclosed herein with substantially zero or first order release
kinetics over an
illustrative (not restrictive) period of, e.g., at most 1 day after
administration, at most 2 days
after administration, at most 3 days after administration, at most 4 days
after administration,
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at most 5 days after administration, or at most 6 days after administration,
at least 7 days after
administration, at least 15 days after administration, at least 30 days after
administration, at
least 45 days after administration, at least 60 days after administration, at
least 75 days after
administration, or at least 90 days after administration.
Micronization
An invention embodiment is a micronized form of a compound(s) and/or
composition(s)
and/or formulation(s) of Formula [X], wherein the meaning of micronization is
well
understood by those of the art. A pharmaceutical composition comprising a
micronized
compound(s) of Formula [X] or a pharmaceutically acceptable salt, solvate,
hydrate or
prodrug thereof, wherein the compound/composition particles have an average
diameter of
less than 30 microns.
Smaller particles have increased surface area to volume ratio, which can
deliver greater water
solubility in the gastrointestinal tract and thence increased bioavailability,
especially useful
for compounds with poor water solubility, wherein many compounds of Formula
[X] have
poor water solubility.
Micronization is the process of reducing the average diameter of a material's
particles. It
usually refers to the reduction of average particle diameters to the
micrometer range, but can
also describe further reduction to the nanometer scale (nanonization). Many
micronization
techniques are based on friction to reduce particle size. Such methods include
one or more of
milling, bashing, grinding, crushing, cutting of larger particles into smaller
particles. More
modern methods use supercritical fluids and fluid energy, to accelerate and
collide particles,
in the micronization process. The most widely applied techniques of
this category include the RESS process (Rapid Expansion of Supercritical
Solutions), the
SAS method (Supercritical Anti-Solvent) and the PGSS method (Particles from
Gas
Saturated Solutions).
Preferably micronized means to reduce to particles that possess an average
diameter of less
than about 30 microns in diameter, preferably less than about 20 microns, more
preferably
less than about 10 microns, and still more preferably less than about 5
microns. The average
diameter (e.g., volume diameter) of the particles is typically at least about
400 nm, preferably
at least about 500 nm, more preferably at least about 700 nm, such as at least
about 800 nm or
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even at least about 1000 nm. An exemplary preferred formulation of the
invention includes
particles having an average diameter of about 0.8 (or 1.0) to about 5.0
microns, such as about
0.8 (or 1.0) to about 2 microns. A micronized drug form promotes better and
more uniform
absorption than forms having a larger average size and a wider distribution of
particle sizes.
In certain embodiments, a large proportion of the particles have diameters
within a defined
range. For example, 80% of the particles have a diameter of from about 0.1
microns to about
20 microns, such as about 0.2 microns to about 10 microns, particularly about
0.2 microns to
about 5 microns or even about 0.2 microns to about 2 microns.
Drugs according to the invention can be micronized using conventional
micronization
equipment, such as the Micron-Master line of micronizers available from The
Jet Pulverizer
Company (Moorestown, N.J.), or processed by a third-party micronization
processor such as
Micron Technologies (Exton, Pa.).
Many drug manufacturing, milling and micronizing machines pulverize substances
into
extremely fine particles, and thus reduce bulk chemicals to the required size
for
pharmaceutical formulation. Particles may also be micronized by chemical or
temperature
controlled processes. The primary benefit to micronizing is the increase in
solubility/bioavailability due to the increase in surface area. These finished
chemicals are
combined and processed further in mixing machines. The mixed ingredients may
then be
mechanically capsulated, pressed into tablets, or made into solutions.
"Micronizing" can be
considered to refer to the processes of making uniform particle size of a
drug, wherein the
size desired may be 10 microns or less, and wherein said process may be
mechanical,
chemical, temperature or pH controlled, or any other process known to those of
the art.
Optimization and control of micronizing processes, particularly relating to
particle size, are
becoming ever more important in the development of pharmaceuticals. Air jet
micronization
is a well proven technique that consistently produces particles in the 1-30
micron range.
Micron Technologies and Jet Pharma are contract micronizers. The primary
advantages of air
jet micronizers are that particle reduction occurs via particle to particle
collisions, with
limited reduction from metal to product contact, and no generation of heat.
Other advantages
include no moving parts and easy to clean surfaces.
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The original principles of jet milling are simple. The powder particles are
fed into the flat
cylindrical milling chamber tangentially through a venturi system by
pressurized air or
nitrogen. The particles are accelerated in a spiral movement inside the
milling chamber by a
number of nozzles placed around the periphery of the chamber. The micronizing
effect takes
place by the collision between the incoming particles and those already
accelerated into the
spiral path. While centrifugal force retains the larger particles at the
periphery of the milling
chamber, the smaller particles exit with the exhaust air from the centre of
the chamber. The
particle size distribution is controlled by adjusting a number of parameters,
two of the main
ones being pressure and feed rate.
In general, there are two types of air jet micronizers or tangential fluid
energy mills, pancake
and loop. The primary difference between the two is in overall distribution.
Loop mills are
excellent choices for cleaning up the tails of the distribution. An additional
advantage is that
both mill types generate no heat. The mills are available in numerous sizes
ranging from 1",
4", 6", 8", 12", and 15" to 20" and provide flexibility in engineering the
desired particle size
in ranges of 1-30 micons.
U.S. Pat. Nos. 6,645,466, 6,623,760, 6,555,135, hereby incorporated by
reference, describe
other micronization procedures.
As used herein, particle size of micronized drug dispersions refers to an
average particle size
as measured by conventional particle size measuring techniques well known to
those skilled
in the art, such as sedimentation field flow fractionation, photon correlation
spectroscopy,
disk centrifugation, and dynamic and static light scattering (e.g., laser
diffraction such as Mie
scattering).
Typically, micronized active compounds are formulated as a freely flowing
liquid (e.g., not a
gel, paste or gum) or as a solid dosage form (e.g., a tablet or capsule). Such
formulations are
preferably safe for internal use (e.g., ingestion, injection). Powders
comprising micronized
drug can be made by spray-drying aqueous dispersions of a micronized drug to
form a dry
powder which consists of aggregated drug particles. Alternatively, the aqueous
dispersion of
drug can contain a dissolved diluent, such as lactose or mannitol, which when
spray dried
forms diluent particles, each of which contains at least one embedded drug
particle.
Micronized drug dispersions can also be freeze dried to obtain powders
suitable for
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formulation into solid dose forms. Such powders comprise aggregated micronized
drug
particles. Freeze dried powders can also be obtained by freeze drying aqueous
dispersions of
drug, which additionally contain a dissolved diluent such as lactose or
mannitol. In these
instances the freeze dried powders consist of particles of diluent, each of
which contains at
least one embedded drug particle.
Other known methods of processing liquid dispersions, and which can be
employed in the
present invention, include granulation, including but not limited to high
shear granulation,
fluid bed granulation, roto granulation, and melt granulation. Additional
methods such as
spray coating and extrusion spherization can also be used. Any other
conventional method for
drying or otherwise processing a liquid dispersion can also be used in the
invention.
The particles are preferably reduced in size at a temperature which does not
significantly
degrade the drug substance. Processing temperatures of less than about 30
degrees to about
40 degrees Centigrade are ordinarily preferred. If desired, the processing
equipment can be
cooled with conventional cooling equipment. The method is conveniently carried
out under
conditions of ambient temperature and at processing pressures which are safe
and effective
for the milling process.
Typically, micronized drug dispersions contain a discrete phase of a drug
substance as
described above having a surface modifier adsorbed on the surface thereof.
Useful surface
modifiers are believed to include those which physiccally adhere to the
surface of the drug
substance but do not chemically bond to the drug. Suitable surface modifiers
can preferably
be selected from known organic and inorganic pharmaceutical excipients. Such
excipients
include various polymers, low molecular weight oligomers, natural products and
surfactants,
such as nonionic and anionic surfactants. Representative examples of
excipients include
gelatin, casein, lecithin (phosphatides), gum acacia, cholesterol, tragacanth,
Stearic acid,
benzalkonium chloride, calcium stearate, glyceryl monostearate, cetostearl
alcohol,
cetomacrogol emulsifying wax, sorbitan esters, polyoxyethylene alkyl ethers,
e.g., macrogol
ethers such as cetomacrogol 1000, polyoxyethylene castor oil derivatives,
polyoxyethylene
sorbitan fatty acid esters, e.g., the commercially available Tweens (Tween
80), polyethylene
glycols, polyoxyethylene stearates, colloidol silicon dioxide, phosphates,
sodium
dodecylsulfate, carboxymethylcellulose calcium, carboxymethylcellulose sodium,
methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylm
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ethycellulose phthalate, noncrystalline cellulose, magnesium aluminum
silicate,
triethanolamine, polyvinyl alcohol, and polyvinylpyrrolidone (PVP). Most of
these excipients
are described in detail in the Handbook of Pharmaceutical Excipients,
published jointly by
the American Pharmaceutical Association and The Pharmaceutical Society of
Great
Britain, the Pharmaceutical Press, 1986, the disclosure of which is hereby
incorporated by
reference in its entirety. The surface modifiers are commercially available
and/or can be
prepared by techniques known in the art.
Nanoparticles
An aspect of the present invention provides a composition that includes
nanoparticles
comprising a compound(s) of Formula [X], or a pharmaceutically acceptable
salt, solvate,
hydrate or prodrug thereof. Such particles can, for example, have a mean
diameter of 50 nm
to 500 nm, such as 100 nm to 200 nm.
Nanoparticles are particles with a diameter from about 5 nm to up to about
1000 nm. The
term "nanoparticles" can refer to particles formed by a polymeric matrix in
which the active
compound is dispersed, also known as "nanospheres, and also refers to
nanoparticles which
are composed of a core containing the active compound which is surrounded by a
polymeric
membrane, also known as "nanocapsules.
Nanoparticles can be prepared by in situ polymerization of dispersed monomers
or by using
preformed polymers. Since polymers prepared in situ are often not
biodegradable and/or
contain toxicological byproducts, nanoparticles from preformed polymers are
preferred.
Nanoparticles from preformed polymers can be prepared by different techniques,
e.g., by
emulsion evaporation, solvent displacement, salting-out and by emulsification
diffusion.
Emulsion evaporation is the classical technique for preparation of
nanoparticles from
preformed polymers. According to this technique, the polymer and the active
compounds are
dissolved in a water-immiscible organic solvent, which is emulsified in an
aqueous solution.
The crude emulsion is then exposed to a high-energy source such as ultrasonic
devices or
passed through high pressure homogenizers or microfluidizers to reduce the
particle size.
Subsequently the organic solvent is removed by heat and/or vacuum resulting in
formation of
the nanoparticles with a diameter of about 100 nm to about 300 nm. Usually,
methylene
chloride and chloroform are used as organic solvent because of their water
insolubility, good
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solubilizing properties, easy emulsification and high volatility. These
solvents are, however,
critical in view of their physiological tolerability. Moreover, the high shear
force needed for
particle size reduction can lead to damage of polymer and/or the active
compound.
The solvent displacement process is described in EPO 274.961 Al. In this
process the active
compound and the polymer are dissolved in an organic solvent which is miscible
with water
in all proportions. This solution is introduced in an aqueous solution
containing a stabilizer
under gentle agitation resulting in spontaneous formation of nanoparticles.
Examples for
suitable organic solvents and stabilizer are acetone or ethanol.
Advantageously chlorinated
solvents and shear stress can be avoided. The mechanism of formation of
nanoparticles has
been explained by interfacial turbulence generated during solvent displacement
(Fessi et al., Int. J. Pharm. 55:R1-R4 (1989)). A solvent displacement
technique was
disclosed by WO 97/03657A1, in which the organic solvent containing the active
compound
and the polymer is introduced into the aqueous solution without agitation.
The salting-out technique is firstly in WO 88/08011A1. In this technique a
solution of a
water-in soluble polymer and an active compound in a water-miscible organic
solvent. Such
as acetone, is mixed with a concentrated aqueous viscous solution or gel
containing a
colloidal stabilizer and a salting-out agent. To the resulting oil-in water
emulsion water is
added in a quantity sufficient to diffuse into the aqueous phase and to induce
rapid diffusion
of the organic solvent into the aqueous phase leading to interfacial
turbulence and formation
of nanoparticles. The organic solvent and the salting-out agent remaining in
the suspension of
nanoparticles are subsequently eliminated by repeated washing with water.
Alternatively, the
solvent and salting-out agent can be eliminated by cross-flow filtration.
In emulsification-diffusion process the polymer is dissolved in a water-
saturated partially
water-soluble organic solvent. This solution is mixed with an aqueous solution
containing a
stabilizer resulting in an oil-in-water emulsion. To this emulsion water is
added causing the
solvent to diffuse into the aqueous external phase accompanied with formation
of
nanoparticles. During particle formation each emulsion droplet leads to
several nanoparticles.
As this phenomenon cannot be fully explained by convection effect caused by
interfacial
turbulence, it has been proposed that diffusion of organic solvent from the
droplets of the
crude emulsion carries molecules of active compound and polymer phase into the
aqueous
phase resulting in supersaturated local regions, from which the polymer
aggregates in the
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form of nanoparticles (Quintanar-Guerrero et al., Colloid. Polym. Sci. 275:640-
647 (1997)).
Advantageously, pharmaceutically acceptable solvents like propylene carbonate
or ethyl
acetate are used as organic solvents.
With the methods described above, nanoparticles can be formed with various
types of
polymers. For use in the method of the present invention, nanoparticles made
from
biocompatible polymers are preferred. The term "biocompatible" refers to
material that after
introduction into a biological environment has no serious effects to the
biological
environment. From biocompatible polymers those polymers are especially
preferred which
are also biodegradable. The term "biodegradable' refers to material that after
introduction into
a biological environment is enzymatically or chemically degraded into smaller
molecules,
which can be eliminated subsequently. Examples are polyesters from
hydroxycarboxylic
acids such as poly(lactic acid) (PLA), poly(glycolic acid) (PGA),
polycaprolactone (PCL),
copolymers of lactic acid and glycolic acid (PLGA), copolymers of lactic acid
and
caprolactone, polyepsilon caprolactone, polyhyroxy butyric acid and
poly(ortho)esters,
polyurethanes, polyanhydrides, polyacetals, polydihydropyrians,
polycyanoacrylates, natural
polymers such as alginate and other polysaccharides including dextran and
cellulose, collagen
and albumin.
Additional methods of preparing nanoparticles include the steps of dispersing
a therapeutic or
diagnostic agent in a liquid dispersion medium and applying mechanical means
in the
presence of grinding media to reduce the particle size of the therapeutic or
diagnostic agent to
an effective average particle size of less than about 400 nm. The particles
can be reduced in
size in the presence of a surface modifier. Alternatively, the particles can
be contacted with a
surface modifier after attrition.
It is preferred, but not essential, that the particle size of the
compound/composition of
Formula [X] be less than about 10 mm as determined by sieve analysis. If the
coarse particle
size is greater than about 100 mm, then it is preferred that the particles be
reduced in size to
less than 100 mm using a conventional milling method such as airjet or
fragmentation
milling.
The compound(s) of Formula [X] can then be added to a liquid medium in which
it is
essentially insoluble to form a premix. The concentration of the therapeutic
or diagnostic
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agent in the liquid medium can vary from about 0.1-99%, and preferably is from
5-30%
(w/w). It is preferred, but not essential, that the surface modifier be
present in the premix.
The concentration of the surface modifier can vary from about 0.1 to about
90%, and
preferably is 1-75%, more preferably 20-60%, by weight based on the total
combined weight
of the compound(s) of Formula [X] and surface modifier. The apparent viscosity
of the
premix suspension is preferably less than about 1000 centipoise.
The premix can be used directly by subjecting it to mechanical means to reduce
the average
particle size in the dispersion to less than 1000 nm. It is preferred that the
premix be used
directly when a ball mill is used for attrition. Alternatively, the
therapeutic or diagnostic
agent and, optionally, the surface modifier, can be dispersed in the liquid
medium using
suitable agitation, e.g., a roller mill or a Cowles type mixer, until a
homogeneous dispersion
is observed in which there are no large agglomerates visible to the naked eye.
It is preferred
that the premix be subjected to such a premilling dispersion step when a
recirculating media
mill is used for attrition.
The mechanical means applied to reduce the particle size of
compound(s)/composition(s) of
Formula [X] conveniently can take the form of a dispersion mill. Suitable
dispersion mills
include a ball mill, an attritor mill, a vibratory mill, and media mills such
as a sand mill and a
bead mill. A media mill is preferred due to the relatively shorter milling
time required to
provide the intended result, desired reduction in particle size. For media
milling, the apparent
viscosity of the premix preferably is from about 100 to about 1000 centipoise.
For ball
milling, the apparent viscosity of the premix preferably is from about 1 to
about 100
centipoise. Such ranges tend to afford an optimal balance between efficient
particle
fragmentation and media erosion.
The attrition time can vary widely and depends primarily upon the particular
mechanical
means and processing conditions selected. For ball mills, processing times of
up to five days
or longer may be required. On the other hand, processing times of less than 1
day (residence
times of one minute up to several hours) have provided the desired results
using a high shear
media mill.
The particles must be reduced in size at a temperature which does not
significantly degrade
the compound(s) of Formula [X]. Processing temperatures of less than about 30-
40 C are
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ordinarily preferred. If desired, the processing equipment can be cooled with
conventional
cooling equipment. The method is conveniently carried out under conditions of
ambient
temperature and at processing pressures which are safe and effective for the
milling process.
For example, ambient processing pressures are typical of ball mills, attritor
mills and
vibratory mills. Control of the temperature, e.g., by jacketing or immersion
of the milling
chamber in ice water are contemplated. Processing pressures from about 1 psi
(0.07 kg/cm2)
up to about 50 psi (3.5 kg/cm2) are contemplated. Processing pressures from
about 10 psi (0.7
kg/cm2) to about 20 psi (1.4 kg/cm2).
The surface modifier, if it was not present in the premix, must be added to
the dispersion after
attrition in an amount as described for the premix above. Thereafter, the
dispersion can be
mixed, e.g., by shaking vigorously. Optionally, the dispersion can be
subjected to a
sonication step, e.g., using an ultrasonic power supply. For example, the
dispersion can be
subjected to ultrasonic energy having a frequency of 20-80 kHz for a time of
about 1 to 120
seconds.
After attrition is completed, the grinding media is separated from the milled
particulate
product (in either a dry or liquid dispersion form) using conventional
separation techniques,
such as by filtration, sieving through a mesh screen, and the like.
In a particular method, a compound(s) of Formula [X] is prepared in the form
of submicron
particles by grinding the agent in the presence of a grinding media having a
mean particle
size of less than about 75 microns.
Another method of forming a nanoparticle dispersion is by microprecipitation.
This is a
method of preparing stable dispersions of a compound(s) of Formula [X] in the
presence of a
surface modifying and colloid stability enhancing surface active agent free of
any toxic
solvents or solubilized heavy metal inpurities by the following procedural
steps:
I. Dissolving the therapeutic or diagnostic agent in aqueous base with
stirring,
2. Adding above #1 formulation with stirring to a surface active surfactant
(or surface
modifiers) solution to form a clear solution, and
3. Neutralizing above formulation #2 with stirring with an appropriate acid
solution. The
procedure can be followed by:
4. Removal of formed salt by dialysis or diafiltration and
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5. Concentration of dispersion by conventional means.
This microprecipitation process produces a dispersion of a compound(s) of
Formula [X] with
Z-average particle diameter less than 400 nm (as measured by photon
correlation
spectroscopy) that is stable in particle size upon keeping under room
temperature or
refrigerated conditions. Such dispersions also demonstrate limited particle
size growth upon
autoclave-decontamination conditions used for standard blood-pool
pharmaceutical agents.
In one embodiment, the above procedure is followed with step 4 which comprises
removing
the formed salts by diafiltration or dialysis. This is done in the case of
dialysis by standard
dialysis equipment and by diafiltration using standard diafiltration equipment
known in the
art. Preferably, the final step is concentration to a desired concentration of
the agent
dispersion. This is done either by diafiltration or evaporation using standard
equipment
known in this art.
In another embodiment of the microprecipitation process, a crystal growth
modifier is used.
A crystal growth modifier is defined as a compound that in the co-
precipitation process
incorporates into the crystal structure of the microprecipitated crystals of
the pharmaceutical
agent, thereby hindering growth or enlargement of the microcrystalline
precipitate, by the so
called Ostwald ripening process. A crystal growth modifier (or a CGM) is a
chemical that is
at least 75% identical in chemical structure to the pharmaceutical agent. By
"identical' is
meant that the structures are identical atom for atom and their connectivity.
Structural identity
is charactarized as having 75% of the chemical structure, on a molecular
weight basis,
identical to the therapeutic or diagnostic agent. The remaining 25% of the
structure may be
absent or replaced by different chemical structure in the CGM. The crystal
growth modifier is
dissolved in step #1 above with the therapeutic or diagnostic agent.
Suitable surface modifiers can preferably be selected from known organic and
inorganic
pharmaceutical excipients. Such excipients include various polymers, low
molecular weight
.. oligomers, natural products and surfactants. Preferred surface modifiers
include nonionic and
ionic surfactants. Representative examples of surface modifiers include
gelatin, casein,
lecithin (phosphatides), gum acacia, cholesterol, tragacanth, Stearic acid,
benzalkonium
chloride, calcium stearate, glycerol monostearate, cetostearyl alcohol,
cetomacrogol
emulsifying wax, sorbitan esters, polyoxyethylene alkyl ethers, e.g., macrogol
ethers such as
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cetomacrogol 1000, polyoxyethylene castor oil derivatives, polyoxyethylene
sorbitan fatty
acid esters, e.g., the commercially available TweensTm, polyethylene glycols,
polyoxyethylene stearates, colloidal silicon dioxide, phosphates, sodium
dodecylsulfate,
carboxymethylcellulose calcium, carboxymethylcellulose sodium,
methylcellulose,
hydroxyethylcellulose, hydroxy propylcellulose, hydroxypropylmethylcellulose
phthalate,
noncrystalline cellulose, magnesium aluminum silicate, triethanolamine,
polyvinyl alcohol,
and polyvinylpyrrolidone (PVP). Most of these surface modifiers are known
pharmaceutical
excipients and are describedin detail in the Handbook of Pharmaceutical
Excipients,
published jointly by the American Pharmaceutical Association and The
Pharmaceutical
Society of Great Britain, the Pharmaceutical Press, 1986.
Particular surface modifiers include polyvinylpyrrolidone, tyloxapol,
poloxamers such as
PluronicsTM F68 and F108, which are block copolymers of ethylene oxide and
propylene
oxide, and polyxamines such as TetronicsTm 908 (also known as Poloxamine 908),
which is a
tetrafunctional block copolymer derived from sequential addition of propylene
oxide and
ethylene oxide to ethylenediamine, available from BASF, dextran, lecithin,
dialkylesters of
sodium sulfosuccinic acid, such as Aerosol OTsTm, which is a dioctylester of
sodium
sulfosuccinic acid, available from American Cyanimid, DuponolsTM P. which is a
sodium
lauryl sulfate, available from DuPont, Tritons Tm X-200, which is an alkylaryl
polyether
sulfonate, available from Rohn and Haas, TweenTm 20 and TweenTm 80, which are
polyoxyethylene sorbitan fatty acid esters, available from ICI Specialty
Chemicals;
CarbowaxsTM 3550 and 934, which are polyethylene glycols available from Union
Carbide:
CrodestasTM F-110, which is a mixture of sucrose stearate and sucrose
distearate, available
from Croda Inc., CrodestasTm SL-40, which is available from Croda, Inc., and
SA9OHCO,
which is C181-137CH2(CON(CH3)CH2(CHOH)4(CH2OH)2. Surface modifiers which have
been
found to be particularly useful include TetronicTm 908, the TweensTm,
PluronicsTM F-68 and
polyvinylpyrrolidone. Other useful surface modifiers include: decanoyl-N-
methylglucamide;
n-decyl-beta-D-glucopyranoside; n-decyl-beta-D-maltopyranoside; n-dodecyl beta-
D-
glucopyranoside; n-dodecyl-beta-D-maltoside; heptanoyl-N-methylglucamide; n-
heptyl-beta-
Dglucopyranoside; n-heptyl-beta-D-thioglucoside; n-hexyl beta-D-
glucopyranoside;
nonanoyl-N-methylglucamide; n-noyl-beta-D-glucopyranoside; octanoyl-N-
methylglucam ide; n-octyl-beta-D-glucopyranoside; octyl beta-D-
thioglucopyranoside; and
the like.
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Another useful surface modifier is tyloxapol (a nonionic liquid polymer of the
alkyl aryl
polyether alcohol type; also known as superinone or triton). Another Surface
modifier is p-
isononylphenoxypoly(glycidol) also known as Olin-10GTm or Surfactant 10-G,
commercially
available as lOGIm from Olin Chemicals, Stamford, Conn.
Two-or more surface modifiers can be used in combination.
Auxiliary surface modifiers can be used to impart resistance to particle
aggregation during
sterilization and include dioctylsulfosuccinate (DOSS), polyethylene glycol,
glycerol, sodium
dodecyl sulfate, dodecyl trimethyl ammonium bromide and a charged phospholipid
such as
dimyristoyl phophatidyl glycerol. Two or more auxiliary surface modifiers can
be used in
combination.
Further description on preparing nanoparticles can be found, for example, in
U.S. Pat. No.
6,264,922, the contents of which are incorporated herein by reference.
Liposomes
Liposomes are a further drug delivery system. Accordingly, in the method of
invention the
active compound(s) can also be administered in the form of a liposome delivery
system.
Liposomes are well-known by a person skilled in the art. Liposomes can be
formed from a
variety of phospholipids, such as cholesterol, stearylamine of
phosphatidylcholines.
Liposomes being usable for the method of invention encompass all types of
liposomes
including, but not limited to, small unilamellar vesicles, large unilamellar
vesicles and
multilamellar vesicles.
= 25
Liposomes are used for a variety of therapeutic purposes, and in particular,
for carrying
therapeutic agents to target cells. Advantageously, liposome-drug formulations
offer the
potential of improved drug-delivery properties, which include, for example,
controlled drug
release. An extended circulation time is often needed for liposomes to reach a
target region,
cell or site. In particular, this is necessary where the target region, cell
or site is not located
near the site of administration. For example, when liposomes are administered
systemically,
it is desirable to coat the liposomes with a hydrophilic agent, for example, a
coating of
hydrophilic polymer chains such as polyethyleneglycol (PEG) to extend the
blood circulation
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lifetime of the liposomes. Such surface-modified liposomes are commonly
referred to as
"long circulating or "sterically stabilized" liposomes.
One surface modification to a liposome is the attachment of PEG chains,
typically having a
molecular weight from about 1000 daltons (Da) to about 5000 Da, and to about 5
mole
percent (%) of the lipids making up the liposomes (see, for example, Stealth
Liposomes, CRC
Press, Lasic. D. and Martin, F., eds., Boca Raton, Fla., (1995)), and the
cited references
therein. The pharmacokinetics exhibited by such liposomes are characterized by
a dose-
independent reduction in uptake of liposomes by the liver and spleen via the
mononuclear
phagocyte system (MPS), and significantly prolonged blood circulation time, as
compared to
non surface-modified liposomes, which tend to be rapidly removed from the
blood and
accumulated in the liver and spleen.
The PEG moiety can have a molecular weight of for example, 750-20,000 Daltons,
such as
1000-10,000 Daltons, in particular 2000-5000 Daltons. In one embodiment, the
complex may
comprise more than one type of PEG moiety (for example, PEG molecular weight
5K and
PEG molecular weight 2K). The PEG moiety may further comprise a suitable
functional
group, such as, for example, methoxy, N-hydroxyl succinimide (NHS),
carbodimide,
etc., for ease of conjugating PEG to the lipid or to the targeting factor.
Table 2 of Harasym et
al. Advanced Drug Delivery Reviews 32:99-118 (1998) provides examples of
suitable
functional groups. Functionalized PEG moieties can be purchased from, for
example,
Shearwater Polymer Inc. (Huntsville, Ala.) and Avanti Polar Lipid Inc.
(Alabaster,
Ala.). In an exemplary embodiment, the PEG moiety is N-[methoxy(polyethylene
glycol)-
5K] (PEG5k). Other types of hydrophilic polymers may be substituted for the
PEG moiety,
including, for example, poloxamer and poloxamine, as described in Feldman et
al. (1997)
Gene Therapy 4(3):189-198: Lemieux et al. (2000) Gene Therapy 7(11):986-91;
Moghimi et
al. (2000) Trends. In Biotechnology 18:412-420; Torchilin (1998) Journal of
Microencapsulation 15(1): 1-19; and Claesson et al. (1996) Colloids &
Surfaces A-Physicochemical & Engineering Aspects 112(2):-3, 131-139.
The PEG moiety may be conjugated to a suitable lipid to form a "pegylated
lipid'. Preferably,
the PEG moiety is covalently attached to the lipid. Suitable lipids include
dioleoylphosphatidyl-ethanolamine (DOPE), cholesterol, and ceramides. Lipids
comprising a
polar end (such as, e.g., phosphatidylethanolamines, including DOPE, DPPE and
DSPE),
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which may be utilized for conjugating to PEG, are preferred for ease of
synthesis of
pegylated lipids. See Harasym et al., Advanced Drug Delivery Reviews 32:99-118
(1998) for non-limiting examples of suitable functionalized lipids. In a
particular
embodiment, the lipid is 1,2-distearoyl-sn-glycero-3-phosphotidylethanolamine
(DSPE) or
dimyristoyl phophatidylethanolamine (DMPE). In a particular embodiment, the
pegylated
lipid comprises 1,2-distearoyl-sn-glycero-3-phosphatidylethanolamine-N-
[methoxy(polyethylene glycol)-51{] (DSPE-PEG5k) or dimyristoyl
phosphatidylethanolamine-N-[methoxy(polyethylene glycol)-5K] (DSPE-PEG5k).
The PEG moiety can be conjugated to the lipid by methods known in the art.
See, for
example, Woodle (1998) Adv. Drug Delivery Reviews 32:139-152 and references
cited
therein; Haselgruber et al. (1995) Bioconjug Chem 6:242-248: Shahinian et al.
(1995)
Biochim Biophys Acta 1239:157-167; Zalipsky etal. (1994) FEBS Lett. 353:71-74;
Zalipsky et al. (1997) Bioconjug Chem. 8(2):111-118: Zalipsky et al. (1995)
Bioconjug
Chem. 6:705-708; Hansen et al. (1995) Biochim Biophys Acta. 1239(2):133-44;
Allen et al.
(1995) Biochim Biophys Acta 1237(2): 99-108; Zalipsky (1995) Bioconjug Chem
6(2): 150-
65; Zalipsky (1993) Bioconjug Chem 4(4): 296-9; and Zalipsky (1995) in Stealth
Liposomes.
(Eds: Lasic, D., et al.) CRC Press, Boca Raton, Fla., p. 93-102. Pegylated
lipids are also
available commercially from, for example, Shearwater Polymer Inc. (Huntsville,
Ala.).
It is to be understood that compounds other than lipids, such as, for example,
peptides,
hydrophobic anchors or polymers, carbohydrates, metals or other ions can be
used for
conjugating with PEG, provided the compounds anchor PEG to the lipid complex,
and allow
PEG to be displayed on the surface of the lipid complex.
While not wishing to be bound by theory, the charge shielding effect provided
by PEG may
enhance the circulatory half-life of the complexes. Shielding may also
increase the resistance
(decrease the sensitivity) of nucleic acid to degradation, for example by
nucleases or other
species present in vitro or in vivo (e.g., hyuralonic acid, poly(Asp)) and/or
decrease or
prevent interactions between individual complex particles or interactions with
other species
present in vitro or in vivo that may lead to increased complex particle size
or aggregation of
complex particles. Accordingly, in a preferred embodiment, the complex
comprises a neutral
surface. In another preferred embodiment, the complex is charge shielded.
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CA 3050553 2019-07-25
As used herein, the term "shielding, and its cognates such as "shielded,
refers to the ability of
"shielding moieties" to reduce the non-specific interaction of the complexes
described herein
with serum complement or with other species present in serum in vitro or in
vivo. Shielding
moieties may decrease the complex interaction with or binding to these species
through one
or more mechanisms, including, for example, non-specific steric or non-
specific electronic
interactions. Examples of such interactions include non-specific electrostatic
interactions,
charge inter actions, Van der Waals interactions, steric-hindrance and the
like. For a moiety
to act as a shielding moiety, the mechanism or mechanisms by which it may
reduce
interaction with, association with or binding to the serum complement or other
species does
not have to be identified. One can determine whether a moiety can act as a
shielding moiety
by determining whether or to what extent a complex binds serum species.
Other moieties that will act as shielding moieties can be identified by their
ability to block
binding of serum complement or the serum complement pathway, such as the C3A
or C5
proteins of the complement pathway. If a moiety is not recognized by (e.g.,
does not bind) at
least one of the components of serum complement or the serum complement
pathway, then
the moiety likely acts as a shielding moiety. In particular examples, if a
moiety does not bind
to or interact with at least one of the C3A or C5 proteins, then the moiety
likely is not bound
by or does not interact with serum complement.
Incorporation of a moiety which does not bind, associate with, or interact
with serum
complement or other serum species on the surface of the complexes described
herein results
in the shielding of the complex. In other words, the components (e.g., lipids)
of the complex
that would be recognized by or would interact with components of serum are
instead shielded
from the serum components (e.g., serum proteins, for example, albumin, serum
complement,
hormones, vitamins, co-factors and others) and therefore are not accessible to
serum
components and thus are not bound by, associated with, or interacting with
these components,
including serum complement. The complex therefore can be described as
"shielded. A moiety
capable of providing shielding can be termed a 'shielding moiety'.
Shielding, as described above, can also be measured by the level of complement
opsonization, as described herein. In particular embodiments, the shielding
moiety will
reduce complement opsonization by approximately 30%, approximately 40%,
approximately
50%, approximately 60%, approximately 65%, approximately 70%, approximately
75%, or
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approximately 80%. In other embodiments, the shielding moiety will reduce
complement
opsonisation by at least 40%, at least 50%, at least 55% or at least 60%.
It should be noted that "shielding moieties' can be multifunctional. For
example, a shielding
moiety may also function as, for example, a targeting factor. A shielding
moiety may also be
referred to as multifunctional with respect to the mechanism(s) by which it
shields the
complex. While not wishing to be limited by proposed mechanism or theory,
examples of
such a multifunctional shielding moiety are pH sensitive endosomal membrane-
disruptive
synthetic polymers, such as PPAA or PEAA. Certain poly(alkylacrylic acids)
have been
shown to disrupt endosomal membranes while leaving the outer cell surface
membrane intact
(Stayton et al. (2000).J. Controll. Release 65:203-220; Murthy et al. (1999)
J. Control!.
Release 61:137-143: W099/34831), thereby increasing cellular bioavailability
and
functioning as a targeting factor. However, PPAA reduces binding of serum
complement to
complexes in which it is incorporated, thus functioning as a shielding moiety.
As will be understood by those of skill in the art, it is important that
incorporation of a
shielding moiety does not eliminate the complex's ability to be delivered to
cells. Therefore,
in some embodiments, complexes incorporating a shielding moiety will further
comprise a
targeting factor. For example, a complex may comprise a cell surface receptor
ligand (e.g.,
folate, an RGD peptide, an LHRH peptide, etc.) that may, for example, be
conjugated to a
lipid or pegylated lipid and optionally also incorporate PPAA. In certain
embodiments, the
lipid-targeting factor conjugate is DSPE-PEG5k-RGD or DSPE-PEG5k-Folate.
The amount or ratio of shielding moiety incorporated in a complex formulation
can be
limited, so as not to eliminate the complex's delivery to cells. Thus in
particular examples,
the complexes comprise less than about 15%, less than about 12%, less than
about 10%, less
than about 8%, less than about 7%, less than about 5%, less than about 4%,
less than about
3%, or less than about 2% shielding moiety. In particular embodiments, the
amount of
shielding moiety is about 10%, about 8%, about 5% or about 2%. A complex may
also
incorporate more than one shielding moiety. In certain embodiments, the amount
of shielding
moiety is at least 2%, at least 5% or at least 8% or at least 10%.
In certain embodiments, the shielding moiety may be conjugated to another
component of the
complex, for example a lipid or pegylated lipid. In certain examples, the
shielding moiety
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CA 3050553 2019-07-25
may be conjugated to a co-lipid or pegylated co-lipid. In other embodiments,
the shielding
moiety is not conjugated to any other component of the complex.
In particular embodiments, the complex is shielded by incorporation of
compounds
.. comprising polyethylene glycol moieties (PEG) or by the incorporation of
synthetic
polymers. In particular examples of the complexes described herein, the
shielded complex
may comprise one or more synthetic polymers, including for example, membrane
disruptive
synthetic polymers, pH sensitive membrane-disruptive synthetic polymers, pH
sensitive
endosomal membrane disruptive synthetic polymers, or poly(alkylacrylic acid)
polymers.
Particular examples of membrane disruptive polymers include poly(alkylacrylic
acid)
polymers such as poly(ethyl acrylic acid) (PEAA) and poly(propyl acrylic acid)
(PPAA).
It is also possible that shielding the complexes may reduce the toxicity of
the complexes.
The pegylated lipid and/or targeting factor-pegylated lipid conjugate and/or
targeting factor-
lipid conjugate may comprise, for example, from about 0.01 to about 30 mol
percent of the
total lipids, more preferably, from about 1 to about 30 mol percent of the
total lipids. The
pegylated lipid and/or targeting factor-pegylated lipid conjugate and/or
targeting factor-lipid
conjugate may comprise, for example, from about 1 to about 20 mol percent,
from about 1 to
about 10 mol percent of the total lipids, from about 2 to about 5 mol percent,
about 1 mol
percent, about 2 mol percent, about 3 mol percent, about 4 mol percent, about
5 mol percent,
about 10 mol percent, about 15 mol percent or about 20 mol percent of the
total lipids. The
complex may comprise a pegylated lipid without conjugated targeting factor as
well as a
targeting factor-pegylated lipid conjugate. The complex may also comprise a
targeting factor-
pegylated lipid conjugate and a targeting factor-lipid conjugate. The complex
may comprise
more than one targeting factor-pegylated lipid conjugate or targeting factor-
lipid conjugate.
The PEG moiety may be the same or different when more than one pegylated lipid
is present
in the complex. In one non limiting example, the targeting factor-pegylated
lipid conjugate
may comprise PEG of 5 KDa molecular weight, and the pegylated lipid without
conjugated
targeting factor may comprise PEG of 750 Da -2 KDa molecular weight. The
complex may
also comprise a pegylated lipid and a targeting factor conjugated to a lipid.
In one
embodiment, the complex comprises a targeting factor-pegylated lipid conjugate
and a
targeting factor-lipid conjugate. Alternatively, in other embodiments, the
complex comprises
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a targeting factor that is not conjugated to lipid or pegylated lipid, and
comprises a pegylated
lipid.
Cyclodextrin
A composition comprising a cyclodextrin(s) and a compound(s) of Formula [X],
or a
pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof,
optionally wherein the
composition is a liquid that comprises at least 30 mM of a compound of Formula
[X],
optionally wherein the cyclodextrin is a substituted cyclodextrin, optionally
wherein the
cyclodextrin is substituted on the 2-, 3- or 6- hydroxyl group of a
glycopyranose moiety,
optionally wherein the cyclodextrin is amorphous, optionally wherein the
cyclodextrin is one
or more of 2-hydroxypropyl-p-cyclodextrin (or derivative
thereof)/hydroxypropyl-P-
cyclodextrin (or derivative thereof)/-cyclodextrin (or derivative thereof)/a-
cyclodextrin (or
derivative thereof)/y-cyclodextrin (or derivative thereof), optionally wherein
the composition
is a lyophilized (e.g. water soluble), optionally wherein the cyclodextrin(s)
is of the structure:
OR (R1)m OR
0
0 0 (R1)m
0 OR RO 0 OR
OR RO
0 OR RO
RO OR RO 0
(R1)m OR OR OR (R1)m
OR 0
0
0
(R1)m OR
RO _n
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or a pharmaceutically acceptable salt, ester, solvate, or hydrate thereof.
Wherein, each R is
independently H, D, halogen, alkyl, alkenyl, alkynyl, cycloalkyl,
heterocycloalkyl, aryl, or
heteroaryl, each of which is optionally substituted; or C(0)ORB, -0C(0)RB, -
C(0)1e, or
C(0)NRARB;
each R1 is selected independently from H, D, alkyl, alkenyl, alkynyl,
cycloalkyl, aryl,
heteroaryl, halogen, hydroxy, amino, CN, CF3, N3, NO2, ORB, SRB, SORB, S02R8, -
N(RB)S(02), RB, -N(RB)S(02)NRARB, NRARB, -C(0)ORB, OC(0)RB, C(0)RB, C(0)NRARB,
or N(RB)C(0)RB; each of which is optionally substituted;
wherein each RA is independently hydrogen, deuterium, alkyl, alkenyl, alkynyl,
cycloalkyl,
heterocycloalkyl, aryl, or heteroaryl, each of which is optionally
substituted;
wherein each RB is independently hydrogen, deuterium, alkyl, alkenyl, alkynyl,
cycloalkyl,
heterocycloalkyl, aryl, or heteroaryl, each of which is optionally
substituted;
wherein n is 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , or 10;
wherein each m is independently 0, 1 , 2 , 3 , 4, or 5.
A way to produce a formulation, particularly a solution, of a compound(s) of
Formula [X] or
a derivative thereof, is through the use of cyclodextrin. By cyclodextrin, it
is meant a-,13 or '-
cyclodextrin. Cyclodextrins are described in detail in Pitha et al., U.S. Pat.
No. 4,727,064,
which is incorporated herein by reference. Cyclodextrins are cyclic oligomers
of glucose;
these compounds form inclusion complexes with any drug whose molecule can fit
into the
lipophile-seeking cavities of the cyclodextrin molecule.
By amorphous cyclodextrin, it is meant non-crystalline mixtures of
cyclodextrins wherein the
mixture is prepared from a-,13-, or y-cyclodextrin. In general the amorphous
cyclodextrin is
prepared by non-selective additions, especially alkylation of the desired
cyclodextrin species.
Reactions are carried out to yield mixtures containing a plurality of
components thereby
preventing crystallization of the cyclodextrin. Various alkylated and
hydroxyalkyl
cyclodextrins can be made and of course will vary, depending upon the starting
species of
cyclodextrin and the addition agent used. Among the amorphous cyclodextrins
suitable for
compositions according to the invention are hydroxypropyl, hydroxyethyl,
glucosyl, maltosyl
and maltotriosyl derivatives of 13-cyclodextrin, carboxyamidomethyl-P-
cyclodextrin,
carboxymethy1-13-cyclodextrin, hydroxypropyl-13-cyclodextrin and diethylamino-
P-
cyclodextrin. The substituted y-cyclodextrins may also be suitable, including
hydroxypropyl,
hydroxyethyl, glucosyl, maltosyl and maltotriosyl derivatives of y-
cyclodextrin.
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The cyclodextrin of the compositions according to the invention may be a-, p-,
or y-
cyclodextrin. a-cyclodextrin contains six glucopyranose units; P-cyclodextrin
contains seven
glucopyranose units; and 'y-cyclodextrin contains eight glucopyranose units.
The molecule is
believed to form a truncated cone having a core opening of 4.7-5.3 angstroms,
6.0-6.5
angstroms, and 7.5-8.3 angstroms in a-, 0-, or y-cyclodextrin respectively.
The composition
according to the invention may comprise a mixture of two or more of the a-, 0-
, or y-
cyclodextrins. Typically, however, the composition according to the invention
will comprise
only one of the a-, 0-, or y-cyclodextrins.
The unmodified a-, 0-, or y-cyclodextrins are less preferred in the
compositions according to
the invention because the unmodified forms tend to crystallize and are
relatively less soluble
in aqueous solutions. More preferred for the compositions according to the
invention are the
a-, 0-, and 'y-cyclodextrins that are chemically modified or substituted.
Chemical substitution
at the 2, 3 and 6 hydroxyl groups of the glucopyranose units of the
cyclodextrin rings yields
increases in solubility of the cyclodextrin compound.
Most preferred cyclodextrins in the compositions according to the invention
are amorphous
cyclodextrin compounds. By amorphous cyclodextrin is meant non-crystalline
mixtures of
cyclodextrins wherein the mixture is prepared from a-, 0-, or y-cyclodextrin.
In general, the
amorphous cyclodextrin is prepared by non-selective alkylation of the desired
cyclodextrin
species. Suitable alkylation agents for this purpose include but are not
limited to propylene
oxide, glycidol, iodoacetamide, chloroacetate, and 2-
diethylaminoethlychloride. Reactions
are carried out to yield mixtures containing a plurality of components thereby
preventing
crystallization of the cyclodextrin. Various alkylated cyclodextrins can be
made and of course
will vary, depending upon the starting species of cyclodextrin and the
alkylating agent used.
Among the amorphous cyclodextrins suitable for compositions according to the
invention
are hydroxypropyl, hydroxyethyl, glucosyl, maltosyl and maltotriosyl
derivatives of 1-
cyclodextrin, carboxyamidom ethyl-0-cyclodextrin, carboxymethyl-P-
cyclodextrin,
hydroxypropyl-P-cyclodextrin and diethylamino-P-cyclodextrin.
Importantly, if the aqueous solution comprising the therapeutic compound(s)
and a
cyclodextrin is to be administered parenterally, especially via the
intravenous route, a
cyclodextrin will be substantially free of pyrogenic contaminants. Various
forms of
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cyclodextrin, such as forms of amorphous cyclodextrin, may be purchased from a
number of
vendors including Sigma-Aldrich, Inc. (St. Louis, Mo., USA). A method for the
production of
hydroxypropy1-13-cyclodextrin is disclosed in Pitha et al., U.S. Pat. No.
4,727,064 which is
incorporated herein by reference.
To produce the formulations according to the invention, a pre-weighed amount
of a
cyclodextrin compound, which is substantially pyrogen free is placed in a
suitable
depyrogenated sterile container. Methods for depyrogenation of containers and
closure
components are well known to those skilled in the art and are fully described
in the United
States Pharmacopeia 23 (United States Pharmacopeial Convention, Rockville, Md.
USA).
Generally, depyrogenation is accomplished by exposing the objects to be
depyrogenated to
temperatures above 400 degrees Centigrade for a period of time sufficient to
fully incinerate
any organic matter. As measured in U.S.P. Bacterial Endotoxin Units, the
formulation will
contain no more than 10 Bacterial Endotoxin Units per gram of amorphous
cyclodextrin. By
substantially pyrogen free is meant that the cyclodextrin con tains less than
10 U.S.P.
bacterial endotoxin units per gram using the U.S.P. method. Preferably, the
cyclodextrin will
contain between 0.1 and 5 U.S.P. bacterial endotoxin units per mg, under
conditions specified
in the United States Pharmacopeia 23.
Sufficient sterile water for injection is added to the substantially pyrogen
free amorphous
cyclodextrin until the desired concentration of the cyclodextrin is in
solution. To this solution
a pre-weighed amount of the therapeutic compound(s), optionally a compound(s)
of Formula
[X], is added with agitation and with additional standing if necessary until
it dissolves.
The solution is then filtered through a sterile 0.22 micron filter into a
sterile holding vessel
and is subsequently filled in sterile depyrogenated vials and is capped. For
products that will
be stored for long periods of time, a pharmaceutically acceptable preservative
may be added
to the solution of therapeutic compound(s) and cyclodextrin prior to
filtration, filling and
capping or alternatively, may be added sterilely after filtration.
As discussed above, the present invention provides improved water soluble
formulations of
compound(s) of Formula [X] and methods of preparing and employing such
formulations.
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The advantages of these water soluble formulations are that a drug is
entrapped in
cyclodextrin in dissolved form. These compositions can be delivered in the
form by slow
infusions or by bolus injection or by other parenteral or oral delivery
routes.
Additional description of the use of cyclodextrin for solubilizing compounds
can be found in
US 2005/0026849, the contents of which are incorporated herein by reference.
In an embodiment, the therapeutic action of a cyclodextrin(s), or
derivative(s) thereof,
synergises with the therapeutic action of one or more compounds of Formula
[X], or a
pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof. For non-
limiting
example, cyclodextrin(s) can slow aging (delay the onset of an age associated
phenotype in a
cell) and a compound of Formula [X] can slow aging (delay the onset of an age
associated
phenotype in a cell) and when co-administered, optionally in a pharmaceutical
composition,
these anti-aging properties add/synergise. A method of treating or delaying
the onset of
geriatric aging of the human or animal body, tissue, or organ comprising
administering a
composition that includes one or more cyclodextrins and one or more compounds
of Formula
[X] to a subject. A method of preventing, treating or delaying the onset of a
disease/condition
associated with aging, and/or a lipofuscin associated disorder, by
administering a
composition that includes one or more of cyclodextrins and one or more
compounds of
Formula [X] to a subject.
Implants
Implantable devices containing a compound(s) of Formula [X] are also included
in the
invention, optionally a bioerodable implant comprising an the active agent
dispersed within a
biodegradable polymer matrix, optionally wherein 75% of the particles of the
active agent
have a diameter of less than about 10 um. The invention provides implantable
(optionally
rechargeable or biodegradable) devices that contain a compound(s) of Formula
[X], or a
pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof. Various
slow release
polymeric devices have been developed and tested in vivo in recent years for
the controlled
delivery of drugs, including proteinacious biopharmaceuticals. A variety of
biocompatible
polymers (including hydrogels), including both biodegradable and non-
degradable polymers,
can be used to form an implant for the sustained release of a drug at a
particular target site.
The biodegradable polymer can be, for example, a poly(lactic-co-glycolic)acid
(PLGA)
copolymer. The ratio of lactic to glycolic acid monomers in the polymer can be
about 50/50
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CA 3050553 2019-07-25
,
weight percentage. Additionally, the PLGA copolymer can be about 20 to about
90 weight
percent of the bioerodible implant. Alternately, the PLGA copolymer can be
about 40 percent
by weight of the bioerodible implant.
In another example, a drug delivery device is formed, in whole or in part, by
co-extruding a
drug core and an outer tube. The outer tube may be permeable, semi-permeable,
or
impermeable to the drug. The drug core may include a polymer matrix which does
not
significantly affect the release rate of the drug. The outer tube, the polymer
matrix of the drug
core, or both may be bioerodible. The co-extruded product can be segmented
into drug
delivery devices. The devices may be left uncoated so that their respective
ends are open, or
the devices may be coated with, for example, a layer that is permeable to the
drug, semi-
permeable to the drug, or bioerodible.
In a further example, a surgically implanted device has a reservoir container
having a
diffusible wall of polyvinyl alcohol or polyvinyl acetate and containing
milligram quantities
of a compound(s) of Formula [X]. As another example, milligram quantities of
agent(s) may
be incorporated into a polymeric matrix, optionally having dimensions of about
2 mm by 4
mm, and made of a polymer such as polycaprolactone, poly(glycolic) acid,
poly(lactic) acid,
or a polyanhydride, or a lipid such as sebacic acid. This is usually
accomplished with the
patient receiving either a topical or local anesthetic and using a small 3-4
mm incision to
enter the implant. The matrix, containing the agent(s), is then inserted
through the incision
and sutured to the sclera using 9-0 nylon.
Additional description of implantable devices can be found, for example, in
U.S. Publication
Nos. 2004/0009222, 2004/0180075, 2005/0048099, 2005/0064010 and 2005/0025810,
the
contents of which are incorporated herein by reference.
Fast melt formulations
Rapid disintegration facilitates delivery of the active material. Rapidly
disintegrating or
dissolving dosage forms are useful for the rapid absorption, particularly
buccal and
sublingual absorption, of pharmaceutically active agents. Fast melt dosage
forms are
beneficial to patients, such as aged and pediatric patients, who have
difficulty in swallowing
typical solid dosage forms, such as caplets and tablets. Additionally, fast
melt dosage forms
circumvent drawbacks associated with, for example, chewable dosage forms,
wherein the
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length of time an active agent remains in a patient's mouth plays an important
role in
determining the amount of taste masking and the extent to which a patient may
object to
throat grittiness of the active agent.
To overcome such problems manufacturers have developed a number of fast melt
solid dose
oral formulations. These are available from manufacturers including Cima Labs,
Fuisz
Technologies Ltd., Prographarmn, R. P. Scherer, Yamanouchi-Shaklee, and McNeil-
PPC,
Inc. All of these manufacturers market different types of rapidly dissolving
solid oral dosage
forms.
Cima Labs markets OraSolTM, which is an effervescent direct compression tablet
having an
oral dissolution time of five to thirty seconds, and DuraSolTM, which is a
direct compression
tablet having a taste-masked active agent and an oral dissolution time of 15
to 45 seconds.
Cima's U.S. Pat. No. 5,607,697, for "Taste Masking Microparticles for Oral
Dosage Forms,"
the contents of which are incorporated herein by reference, describes a solid
dosage form
consisting of coated microparticles that disintegrate in the mouth. The
microparticle core of
Cima's patented oral dosage form has a pharmaceutical agent and one or more
sweet-tasting
compounds having a negative heat of solution wherein the sweet-tasting
compound can be
mannitol, sorbitol, a mixture of an artificial sweetener and menthol, a
mixture of sugar and
menthol, or methyl salicylate. The microparticle core is coated, at least
partially, with a
material that retards dissolution in the mouth and masks the taste of the
pharmaceutical agent.
The microparticles are then compressed to form a tablet. Cima's patent
discloses that other
excipients can also be added to the tablet formulation.
WO 98/46215 for "Rapidly Dissolving Robust Dosage Form," the contents of which
are
incorporated herein by reference, is directed to a hard, compressed, fast melt
formulation
having an active ingredient and a matrix of at least a non-direct compression
filler and
lubricant. A non-direct compression filler is typically not free-flowing, in
contrast to a direct
compression (DC grade) filler, and usually requires additionally processing to
form free-
flowing granules.
Cima also has U.S. patents and international patent applications directed to
effervescent
dosage forms (U.S. Pat. Nos. 5,503,846, 5,223,264, and 5,178,878, the contents
of each are
incorporated herein by reference) and tableting aids for rapidly dissolving
dosage forms (U.S.
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Pat. Nos. 5,401,513 and 5,219,574, the contents of both are incorporated
herein by reference),
and rapidly dissolving dosage forms for water soluble drugs (WO 98/14179 for
"Taste-
Masked Microcapsule Composition and Methods of Manufacture", the contents of
which are
incorporated herein by reference).
Fuisz Technologies, now part of BioVail, markets Flash DoseTM, which is a
direct
compression tablet containing a processed excipient called ShearformTM.
ShearformTM is a
cotton candy-like substance of mixed polysaccharides converted to amorphous
fibers. U.S.
patents describing this technology include U.S. Pat. No. 5,871,781 for
"Apparatus for Making
Rapidly Dissolving Dosage Units;" U.S. Pat. No. 5,869,098 for "Fast-Dissolving
Comestible
Units Formed Under High-Speed/High-Pressure Conditions;" U.S. Pat. Nos.
5,866,163,
5,851,553, and 5,622,719, all for "Process and Apparatus for Making Rapidly
Dissolving
Dosage Units and Product Therefrom;" U.S. Pat. No. 5,567,439 for "Delivery of
Controlled-
Release Systems;" and U.S. Pat. No. 5,587,172 for "Process for Forming Quickly
Dispersing
Comestible Unit and Product Therefrom," each of which is incorporated herein
by reference.
Prographarm markets FlashtabTM, which is a fast melt tablet having a
disintegrating agent
such as carboxymethyl cellulose, a swelling agent such as a modified starch,
and a taste-
masked active agent. The tablets have an oral disintegration time of under one
minute (U.S.
Pat. No. 5,464,632, the contents of which are incorporated herein by
reference).
R. P. Scherer markets ZydisTM, which is a freeze-dried tablet having an oral
dissolution time
of 2 to 5 seconds. Lyophilized tablets are costly to manufacture and difficult
to package
because of the tablets sensitivity to moisture and temperature. U.S. Pat. No.
4,642,903 (R. P.
Scherer Corp.), the contents of which are incorporated herein by reference,
refers to a fast
melt dosage formulation prepared by dispersing a gas throughout a solution or
suspension to
be freeze-dried. U.S. Pat. No. 5,188,825 (R. P. Scherer Corp.), the contents
of which are
incorporated herein by reference, refers to freeze-dried dosage forms prepared
by bonding or
complexing a water-soluble active agent to or with an ion exchange resin to
form a
substantially water insoluble complex, which is then mixed with an appropriate
carrier and
freeze dried. U.S. Pat. No. 5,631,023 (R. P. Scherer Corp.), the contents of
which are
incorporated herein by reference, refers to freeze-dried drug dosage forms
made by adding
xanthan gum to a suspension of gelatin and active agent. Finally, U.S. Pat.
No. 5,827,541 (R.
P. Scherer Corp.), the contents of which are incorporated herein by reference,
discloses a
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process for preparing solid pharmaceutical dosage forms of hydrophobic
substances. The
process involves freeze-drying a dispersion containing a hydrophobic active
ingredient and a
surfactant, in a non-aqueous phase; and a carrier material, in an aqueous
phase.
Yamanouchi-Shaklee markets WowtabTM, which is a tablet having a combination of
a low
moldability and a high moldability saccharide. U.S. patents covering this
technology include
U.S. Pat. No. 5,576,014 for "Intrabuccally Dissolving Compressed Moldings and
Production
Process Thereof," and U.S. Pat. No. 5,446,464 for "Intrabuccally
Disintegrating Preparation
and Production Thereof," both of which are incorporated herein by reference.
Other companies owning rapidly dissolving technology include Janssen
Pharmaceutica. U.S.
patents assigned to Janssen describe rapidly dissolving tablets having two
polypeptide (or
gelatin) components and a bulking agent, wherein the two components-have a net
charge of
the same sign, and the first component is more soluble in aqueous solution
than the second
component. See U.S. Pat. No. 5,807,576 for "Rapidly Dissolving Tablet;" U.S.
Pat. No.
5,635,210 for "Method of Making a Rapidly Dissolving Tablet;" U.S. Pat. No.
5,595,761 for
"Particulate Support Matrix for Making a Rapidly Dissolving Tablet;" U.S. Pat.
No.
5,587,180 for "Process for Making a Particulate Support Matrix for Making a
Rapidly
Dissolving Tablet;" and U.S. Pat. No. 5,776,491 for "Rapidly Dissolving Dosage
Form,"
each of which is incorporated herein by reference.
Eurand America, Inc. has U.S. patents directed to a rapidly dissolving
effervescent
composition having a mixture of sodium bicarbonate, citric acid, and
ethylcellulose (U.S. Pat.
Nos. 5,639,475 and 5,709,886, the contents of which are incorporated herein by
reference).
L.A.B. Pharmaceutical Research owns U.S. patents directed to effervescent-
based rapidly
dissolving formulations having a pharmaceutically active ingredient and an
effervescent
couple comprising an effervescent acid and an effervescent base (U.S. Pat.
Nos. 5,807,578
and 5,807,577, each of which is incorporated herein by reference).
Schering Corporation has technology relating to buccal tablets having an
active agent, an
excipient (which can be a surfactant) or at least one of sucrose, lactose, or
sorbitol, and either
magnesium stearate or sodium dodecyl sulfate (U.S. Pat. Nos. 5,112,616 and
5,073,374, each
of which is incorporated herein by reference).
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Laboratoire L. LaFon owns technology directed to conventional dosage forms
made by
lyophilization of an oil-in-water emulsion in which at least one of the two
phases contains a
surfactant (U.S. Pat. No. 4,616,047, the contents of which are incorporated
herein by
reference). For this type of formulation, the active ingredient is maintained
in a frozen
suspension state and is tableted without micronization or compression, as such
processes
could damage the active agent.
Takeda Chemicals Inc., Ltd. owns technology directed to a method of making a
fast
dissolving tablet in which an active agent and a moistened, soluble
carbohydrate are
compression molded into a tablet, followed by drying of the tablets (U.S. Pat.
No. 5,501,861,
which is incorporated herein by reference).
Finally, Elan's U.S. Pat. No. 6,316,029, for "Rapidly Disintegrating Oral
Dosage Form," the
contents of which are incorporated by reference, discloses fast melt dosage
forms comprising
nanoparticulate active agents.
In one example of fast melt tablet preparation, granules for fast melt tablets
made by either
the spray drying or pre-compacting processes are mixed with excipients and
compressed into
tablets using conventional tablet making machinery. The granules can be
combined with a
variety of carriers including low density, high moldability saccharides, low
moldability
saccharides, polyol combinations, and then directly compressed into a tablet
that exhibits an
improved dissolution and disintegration profile.
The granules used to make the tablets can be, for example, mixtures of low
density alkali
earth metal salts or carbohydrates. For example, a mixture of alkali earth
metal salts includes
a combination of calcium carbonate and magnesium hydroxide. Similarly, a fast
melt tablet
can be prepared according to the methods of the present invention that
incorporates the use of
A) spray dried extra light calcium carbonate/maltodextrin, B) magnesium
hydroxide and C) a
eutectic polyol combination including sorbitol instant, xylitol and mannitol.
These materials
can be combined to produce a low density tablet that dissolves very readily
and promotes the
fast disintegration of the active ingredient. Additionally, the pre-compacted
and spray dried
granules can be combined in the same tablet.
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For fast melt tablet preparation, a compound(s) useful in the present
invention can be in a
form such as solid, particulate, granular, crystalline, oily or solution. The
compound(s) for
use in the present invention may be a spray dried product or an adsorbate that
has been pre-
compacted to a harder granular form that reduces the medicament taste. A
pharmaceutical
active ingredient for use in the present invention may be spray dried with a
carrier that
prevents the active ingredient from being easily extracted from the tablet
when chewed.
In addition to being directly added to the tablets of the present invention,
the medicament
drug itself can be processed by the pre-compaction process to achieve an
increased density
prior to being incorporated into the formulation.
The pre-compaction process used in the present invention can be used to
deliver poorly
soluble pharmaceutical materials so as to improve the release of such
pharmaceutical
materials over traditional dosage forms. This could allow for the use of lower
dosage levels to
deliver equivalent bioavailable levels of drug and thereby lower toxicity
levels. Poorly
soluble pharmaceutical materials can be used in the form of nanoparticles,
which are
nanometer-sized particles.
In addition to the active ingredient and the granules prepared from low
density alkali earth
metal salts and/or water soluble carbohydrates, the fast melt tablets can be
formulated using
conventional carriers or excipients and well established pharmaceutical
techniques.
Conventional carriers or excipients include, but are not limited to, diluents,
binders, adhesives
(i.e., cellulose derivatives and acrylic derivatives), lubricants (i.e.,
magnesium or calcium
stearate, vegetable oils, polyethylene glycols, talc, sodium lauryl sulphate,
polyoxy ethylene
monostearate), disintegrants, colorants, flavorings, preservatives, sweeteners
and
miscellaneous materials such as buffers and adsorbents.
Additional description of the preparation of fast melt tablets can be found,
for example, in
U.S. Pat. No. 5,939,091, the contents of which are incorporated herein by
reference.
Animal feed
Componentry to this invention is to add a compound(s) of Formula [X] to animal
feed and/or
drinking water/fluid.
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The addition of the active compound of the invention to animal feed is
preferably
accomplished by preparing an appropriate feed premix containing the active
compound in an
effective amount and incorporating the premix into the complete ration.
Alternatively, an
intermediate concentrate or feed supplement containing the active ingredient
can be blended
into the feed. The way in which such feed premixes and complete rations can be
prepared and
administered are described in reference books (such as "Applied Animal
Nutrition", W.H.
Freedman and CO., San Francisco, U.S.A., 1969 or "Livestock Feeds and Feeding"
0 and B
books, Corvallis, Oreg., U.S.A., 1977).
Cycling Therapy
In certain embodiments, the prophylactic/therapeutic agents provided herein
are cyclically
administered to a subject. Cycling therapy involves the administration of an
active agent for a
period of time, followed by a rest for a period of time, wherein the rest
period equals a day,
days, weeks, or months without the compound(s) being administered, before the
administration period starts again, and this cycle is repeated over a period
of time, optionally
wherein the length (and dosage) of the administration and/or rest periods can
be modulated,
optionally to find the best cycle for the subject. A variant is wherein the
rest period isn't
complete rest, but the compound(s) is administered at lower dosage. Cycling
therapy can
reduce the development of resistance to one or more of the therapies, avoid,
or reduce the
side effects of one of the therapies, and/or improves the efficacy of the
treatment. In an
embodiment, a cycle of administration with one or more compounds of Formula
[X] is
implemented completely out of phase, or with a degree of overlap, with a cycle
of
administration of one or more other compounds of Formula [X], or another drug
approved for
human use by the regulatory body applicable to the country that the subject
resides e.g. the
FDA applies for a subject in the USA.
Optimising for brain/CNS administration
Delivery approaches can be used to deliver therapeutic agents to the brain
whilst
circumventing the blood-brain barrier. Such approaches utilize intrathecal
injections, surgical
implants (Ommaya, Cancer Drug Delivery, 1: 169-178 (1984) and U.S. Pat. No.
5,222,982),
interstitial infusion (Bobo et al., Proc. Natl. Acad. Sci. U.S.A., 91: 2076-
2080 (1994)), and
the like. These strategies deliver an agent to the CNS by direct
administration into the
cerebrospinal fluid (CSF) or into the brain parenchyma (ECF).
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Drug delivery to the central nervous system through the cerebrospinal fluid
can be achieved,
for example, by means of a subdurally implantable device the "Ommaya
reservoir". The drug
is injected into the device and subsequently released into the cerebrospinal
fluid surrounding
the brain. It can be directed toward specific areas of exposed brain tissue
which then adsorb
the drug. This adsorption is limited since the drug does not travel freely. A
modified device,
whereby the reservoir is implanted in the abdominal cavity and the injected
drug is
transported by cerebrospinal fluid (taken from and returned to the spine) to
the ventricular
space of the brain, is used for agent administration. Through omega-3
derivatization, site-
specific biomolecular complexes can overcome the limited adsorption and
movement of
therapeutic agents through brain tissue.
Another strategy to improve agent delivery to the CNS is by increasing the
agent absorption
(adsorption and transport) through the blood-brain barrier and the uptake of
therapeutic agent
by the cells (Broadwell, Acta Neuropathol., 79: 117-128 (1989); Pardridge
etal., J.
Pharmacol. Experim. Therapeutics, 255: 893-899 (1990); Banks et al., Progress
in Brain
Research, 91: 139-148 (1992); Pardridge, Fuel Homeostasis and the Nervous
System, ed.:
Vranic et al., Plenum Press, New York, 43-53 (1991)). The passage of agents
through the
blood-brain barrier to the brain can be enhanced by improving either the
permeability of the
agent itself or by altering the characteristics of the blood-brain barrier.
Thus, the passage of
the agent can be facilitated by increasing its lipid solubility through
chemical modification,
and/or by its coupling to a cationic carrier, or by its covalent coupling to a
peptide vector
capable of transporting the agent through the blood-brain barrier. Peptide
transport vectors
are also known as blood-brain barrier perrneabilizer compounds (U.S. Pat. No.
5,268,164).
Site specific macromolecules with lipophilic characteristics useful for
delivery to the brain
are described in U.S. Pat. No. 6,005,004. Other examples (U.S. Pat. No.
4,701,521, and U.S.
Pat. No. 4,847,240) describe a method of covalently bonding an agent to a
cationic
macromolecular carrier which enters into the cells at relatively higher rates.
These patents
teach enhancement in cellular uptake of bio-molecules into the cells when
covalently bonded
to cationic resins. U.S. Pat. No. 4,046,722 discloses anti-cancer drugs
covalently bonded to
cationic polymers for the purpose of directing them to cells bearing specific
antigens. The
polymeric carriers have molecular weights of about 5,000 to 500,000. Such
polymeric
carriers can be employed to deliver compounds described herein in a targeted
manner.
Further work involving covalent bonding of an agent to a cationic polymer
through an acid-
sensitive intermediate (also known as a spacer) molecule, is described in U.S.
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Pat. No. 4,631,190 and U.S. Pat. No. 5,144,011. Various spacer molecules, such
as cis-
aconitic acid, are covalently linked to the agent and to the polymeric
carrier. They control the
release of the agent from the macromolecular carrier when subjected to a mild
increase in
acidity, such as probably occurs within a lysosome of the cell. The drug can
be selectively
hydrolyzed from the molecular conjugate and released in the cell in its
unmodified and active
form. Molecular conjugates are transported to lysosomes, where they are
metabolized under
the action of lysosomal enzymes at a substantially more acidic pH than other
compartments
or fluids within a cell or body. The pH of a lysosome is shown to be about
4.8, while during
the initial stage of the conjugate digestion, the pH is possibly as low as
3.8.
Kits
The present invention also encompasses an article of manufacture. As used
herein, article of
manufacture is intended to include, but not be limited to, kits and packages.
The kit may
comprise multiple internal containers to keep components separate. An article
of manufacture
of the present invention, comprises: (a) a first container (most preferably
sterilised,
sterilization methodology well known to those skilled in the art, and
packaged/sealed to
maintain sterilization; optionally it is a blister pack, wherein this term and
its meaning is well
known in the art); (b) a pharmaceutical composition located within the first
container,
wherein the composition, comprises: a first therapeutic agent, comprising: a
compound of the
present invention or a pharmaceutically acceptable salt form thereof; and, (c)
a package insert
stating that the pharmaceutical composition can be used for the treatment of a
specified
disease(s)/disorder(s)/condition(s) in a subject, optionally stating the
treatment of a
disease(s)/disorder(s)/condition(s) refered to herein, optionally stating the
treatment of
cancer. In another embodiment, the package insert states that the
pharmaceutical composition
can be used in combination (as defined previously) with a second therapeutic
agent(s) to treat
the same specified disease(s)/disorder(s) in a subject, optionally wherein
this is cancer. The
article of manufacture can further comprise: (d) a second container, wherein
components (a)
and (b) are located within the second container and component (c) is located
within or outside
of the second container. Located within the first and second containers means
that the
respective container holds the item within its boundaries.
The first container is a receptacle used to hold a pharmaceutical composition.
This
container can be for manufacturing, storing, shipping, and/or individual/bulk
selling. First
container is intended to cover a bottle, jar, vial, flask, syringe, tube
(e.g., for a cream
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preparation), or any other container used to manufacture, hold, store, or
distribute a
pharmaceutical product.
The second container is one used to hold the first container and, optionally,
the package
insert. Examples of the second container include, but are not limited to,
boxes (e.g.,
cardboard or plastic), crates, cartons, bags (e.g., paper or plastic bags),
pouches, and sacks.
The package insert can be physically attached to the outside of the first
container via tape,
glue, staple, or another method of attachment, or it can rest inside the
second container
without any physical means of attachment to the first container.
Alternatively, the package
insert is located on the outside of the second container. When located on the
outside of the
second container, it is preferable that the package insert is physically
attached via tape, glue,
staple, or another method of attachment. Alternatively, it can be adjacent to
or touching the
outside of the second container without being physically attached.
The package insert is a label, tag, marker, etc. that recites information
relating to the
pharmaceutical composition located within the first container. The information
recited will
usually be determined by the regulatory agency governing the area in which the
article of
manufacture is to be sold (e.g., the United States Food and Drug
Administration). Preferably,
the package insert specifically recites the indications for which the
pharmaceutical
composition has been approved. The package insert may be made of any material
on which a
person can read information contained therein or thereon. Preferably, the
package insert is a
printable material (e.g., paper, plastic, cardboard, foil, adhesive-backed
paper or plastic, etc.)
on which the desired information has been formed (e.g., printed or applied).
Information may
be provided by reference to a website(s).
Herein disclosed are articles of manufacture that include a compound(s) and/or
pharmaceutical composition(s) described herein in a suitable container. The
container may be
a vial, jar, ampoule, preloaded syringe, intravenous bag or other used for
containing a drug
for human or veterinary use. Disclosed herein are kits that include a
compound(s) of the
disclosure, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug
thereof, and
suitable packaging, optionally with a package insert, optionally a paper
package insert. In
some embodiments, a kit further includes instructions for use, which can be
upon a package
insert. In some embodiments, a kit includes a compound(s) of the disclosure,
or a
pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof,
packaging and a label
and/or instructions for use of the compound(s)/composition(s) in the
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treatment/amelioration/prevention/combating of indications, optionally
including one or more
diseases/disorders/conditions described herein. Pharmaceutical
compound(s)/composition(s)
of this invention can be inside one or more embodiments of a
container/pack/package/dispenser, optionally together with instructions for
administration,
optionally with drug information such as, for example,
disease(s)/disorder(s)/condition(s)
treated/ameliorated/prevented/combated, possible side-effects, desirable or
undesirable/dangerous drug interactions possible in the subject, information
related to what to
do upon overdose, and any information specified for inclusion according to the
relevant
regulatory authority e.g. the FDA in the USA, and optionally wherein each dose
(e.g. daily)
of compound(s)/composition(s) is compartmentalized in its own compartment or
discrete
storage within a bigger package, optionally labelled (e.g. with day of week).
For example, a
packaged product may comprise a container; an effective amount of a
compound(s) of the
invention; and an insert associated with the container, indicating
administering the
compound(s) for treating a disorder(s), optionally cancer. The composition(s)
of the kit may
be provided as any suitable form. The kits may contain instructions for
mixing, diluting,
and/or administrating the compounds. In certain embodiments, the kit provided
herein further
comprises a device that is used to administer the active ingredients. Examples
of such devices
include, but are not limited to, syringes, drip bags, patches, and inhalers.
The kits also can
include other container(s) with pharmaceutically acceptable vehicle(s) that
can be used to
administer one or more active ingredients, such as one or more solvents,
surfactants,
preservatives, and/or diluents (e.g., normal saline (0.9% NaCI), or 5%
dextrose) as well as
containers for mixing, diluting or administering the components to the subject
in need of such
treatment. When the composition provided is a dry powder, the powder may be
reconstituted
by the addition of a suitable solvent, which may also be provided. In
embodiments where
liquid forms of the composition are sued, the liquid form may be concentrated
or ready to
use. The solvent will depend on the compound and the mode of use or
administration.
Suitable solvents for drug compositions are well known and are available in
the literature. In
a particular embodiment, the solution for administering the compound is
sterilised. In some
embodiments, the package insert instructs a user of the kit to administer the
compound or
pharmaceutical composition to a subject. In some embodiments, the package
insert instructs a
user of the kit to mix the compound or pharmaceutical composition with an
aqueous solution.
In some embodiments, the package insert instructs a user of the kit to orally
administer the
compound to the subject. The kits may further comprise conventional
pharmaceutical kit
components which will be readily apparent to those skilled in the art.
Optionally the kit
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contains a compound(s) of Formula [X] and a further compound or compounds
approved for
human use by the FDA and/or EMA, for example an anti-cancer drug(s), and
optionally
instructions for administrating the compounds contained therein to a subject,
optionally with
materials to perform or assist said administration.
An article of manufacture embodiment of this invention is almitrine dimesylate
in a kit for
cancer treatment wherein almitrine dimesylate, or some other composition of
almitrine, is in a
package associated with instructions for, and/or information pertaining to,
its use as an anti-
cancer treatment in a subject. These instructions can be inside the package,
optionally in an
insert, or outside the package and distributed with the package, most
preferably physically
attached to the package, for example being on a label adhered to the package.
Promotion; business methods of the invention
The invention encompasses methods of promoting the
treatment/amelioration/prevention/combating of a
disease/disorder/condition/etiology/unwanted body feature or aspect by a
subject taking or
being administered a compound(s) and/or pharmaceutical composition(s) of the
present
invention. The invention encompasses a method of promoting the
treatment/amelioration/prevention/combating of cancer/tumor/abnormal cell
proliferation
according to any of the methods/compounds/compositions/kits/combinations
described herein
e.g. promoting the administration of a therapeutically effective amount of a
compound(s) of
this invention to the subject in need thereof, optionally where the subject
has, or is suspected
of having, or could, have cancer. As used herein, "promoted" or "promoting"
includes all
methods of doing business including methods of education, hospital and other
clinical
instruction, pharmaceutical industry activity including pharmaceutical
sales/licensing, and
any advertising/marketing or other promotional activity (directed at
patient(s) and/or clinical
professional(s)) including written, oral and electronic communication of any
form, associated
with compounds/compositions/methods/kits of the invention optionally in
connection with
treatment of cell proliferation, cancers or tumors. "Instructions" can define
a component of
promotion, and typically involve written instructions on or associated with
packaging of
compounds/compositions of the invention. Instructions also can include any
oral or electronic
instructions provided in any manner. The "kit" typically defines a package
including any one
or a combination of the compounds/compositions of the invention and the
instructions, but
can also include the composition of the invention and instructions of any form
that are
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provided in connection with the composition in a manner such that a clinical
professional will
clearly recognize that the instructions are to be associated with the specific
compound(s)/composition(s).
An invention embodiment is the promotion of almitrine dimesylate, or some
composition(s)
of almitrine, for treatment/amelioration/prevention/combat of cancer in a
subject. A method
of this invention is wherein almitrine or almitrine dimesylate, or some other
composition(s)
of almitrine, is exchanged between people, either directly or by exchange(s)
via further proxy
person(s), for example between physician and patient, or physician and
almitrine distributor,
or pharmaceutical company representative and patient via almitrine distributor
and physician,
and associated with this physical exchange of almitrine/almitrine composition
between
people, information is also transmitted between these people, optionally
orally or written,
pertaining to the use of almitrine and/or almitrine composition(s) for anti-
cancer use in a
subject.
An invention embodiment is a business method wherein the subject pays for an
insurance
product from a company (e.g. an insurance company/medical insurance company),
optionally
paying a daily, weekly, monthly or yearly fee, and when the subject is need of
a compound(s)
of this invention, optionally as directed by a physician or medical/clinical
practitioner, the
insurance company pays for all or some of its cost, or the subject and/or the
medical practice
(a place of one or more working clinical professionals) pays for it and this
cost to the subject
and/or medical practice is then reimbursed to the subject and/or medical
practice, either in
part or in total, by the insurance company, optionally wherein the insurance
covers the cost of
other medical treatment also. Put another way, the cost of the compound(s) of
this invention
is not met by the subject to be treated directly but through one or more
intermediates which
can include a hospital or clinical establishment and an insurance company or
companies,
wherein the subject pays an insurance company for a promise/agreement/contact
that the
company will pay directly or ultimately for a compound(s) of this invention
should the
subject require it for their treatment.
Clinical Trials
Embodied by this invention are methods of testing/assessing/verifying the
safety and/or
therapeutic/beneficial activity of a compound(s) and/or pharmaceutical
composition(s) of this
invention in a subject, optionally in human subjects using a clinical trial
methodology or
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methodologies, many variants of which are known to, and could be conceived by,
those of
ordinary skill of the art, optionally wherein the human subjects have cancer,
optionally
wherein this clinical trial(s) is to obtain regulatory approval for a human
use(s) from a
regulatory body(s), optionally a national or supranational regulator,
optionally the FDA in
USA, optionally wherein the clinical trial is for a compound(s)/composition(s)
of this
invention in use for anti-cancer treatment/amelioration/prevention/combat in a
subject(s),
optionally administered with another drug(s)/treatment(s), optionally wherein
this other
drug(s)/treatment(s) is also for anti-cancer treatment, optionally wherein it
already has
regulatory approval (e.g. by FDA) for this use or other. Possible clinical
trial endpoints
include, illustratively, not restrictively, disease free survival (DFS),
objective response rate
(ORR), time to progression (TTP), progression-free survival (PFS), and time-to-
treatment
failure (TTF), where these terms are well known in the art. Stable disease or
lack thereof is
determined by methods known in the art such as evaluation of patient symptoms,
physical
examination, visualization of the tumor that has been imaged using X-ray, CAT,
PET, or Mill
scan and other commonly accepted evaluation modalities.
Solvents
The reactions of the synthetic methods described herein can be carried out in
suitable solvents
which can be readily selected by one skilled in the art of organic synthesis.
Generally,
suitable Solvents are solvents which are substantially non-reactive with the
starting materials
(reactants), the intermediates, or products at the temperatures at which the
reactions are
carried out, i.e., temperatures which can range from the solvents freezing
temperature to the
solvent's boiling temperature. A given reaction can be carried out in one
solvent or a mixture
of more than one solvent. Depending on the particular reaction, suitable
solvents for a
particular work-up following the reaction can be selected. Suitable solvents,
as used herein
can include, by way of example and without limitation, chlorinated solvents,
hydrocarbon
solvents, aromatic solvents, ether solvents, protic solvents, polar aprotic
solvents, and
mixtures thereof.
Suitable halogenated solvents include, but are not limited to, carbon
tetrachloride,
bromodichloromethane, dibromochloromethane, bromoform, chloroform,
bromochloromethane, dibromomethane, butyl chloride, dichloromethane,
tetrachloroethylene,
trichloroethylene, 1,1,1-trichloroethane, 1,1,2-trichloroethane, 1,1-
dichloroethane, 2-
chloropropane, hexafluorobenzene, 1,2,4-trichlorobenzene, 0-dichlorobenzene,
chlorobenzene, fluorobenzene, fluorotrichloromethane, chlorotrifluoromethane,
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CA 3050553 2019-07-25
bromotrifluoromethane, carbon tetrafluoride, dichlorofluoromethane,
chlorodifluoromethane,
trifluoromethane, 1,2-dichlorotetrafluorethane and hexafluoroethane.
Suitable hydrocarbon solvents include, but are not limited to, alkane or
aromatic solvents
such as cyclohexane, pentane, hexane, toluene, cycloheptane,
methylcyclohexane, heptane,
ethylbenzene, m-, o-, or p-Xylene, octane, indane, nonane, benzene,
ethylbenzene, and m-, o-
, or p-xylene.
Suitable ether solvents include, but are not limited to dimethoxymethane,
tetrahydrofuran,
1,3-dioxane, 1,4-dioxane, furan, diethyl ether, ethylene glycol dimethyl
ether, ethylene glycol
diethyl ether, diethylene glycol dimethylether, diethylene glycol diethyl
ether, triethylene
glycol diisopropyl ether, anisole, or t-butyl methyl ether.
Suitable protic solvents include, but are not limited to water, methanol,
ethanol, 2-
nitroethanol, 2-fluoroethanol, 2.2.2-trifluoroethanol, ethylene glycol, 1-
propanol, 2-propanol,
2-methoxyethanol, 1-butanol, 2-butanol, 1-butyl alcohol, t-butyl alcohol, 2-
ethoxyethanol,
diethylene glycol, 1-, 2-, or 3-pentanol, neo-pentyl alcohol, t-pentyl
alcohol, diethylene
glycol monomethyl ether, diethylene glycol monoethyl ether, cyclohexanol,
benzyl alcohol,
phenol, and glycerol.
Suitable aprotic solvents include, but are not limited todimethylformamide
(DMF),
dimethylacetamide (DMAC), 1,3-dimethy1-3,4,5,6-tetrahydro-2(1H)-pyrimidinone
(DMPU), 1,3-dimethy1-2-imidazolidinone (DMI), N-methylpyrrolidinone (NMP),
formamide, N-methylacetamide,N-methylformamide, acetonitrile (ACN),
dimethylsulfoxide
(DMSO), propionitrile, ethyl formate, methyl acetate, hexachloroacetone,
acetone, ethyl
methyl ketone, ethyl acetate, isopropyl acetate, t-butyl acetate, Sulfolane,
N,Ndimethylpropionamide,nitromethane, nitrobenzene, and
hexamethylphosphoramide.
Polymorphs
In certain embodiments, a compound(s) of Formula [X] is a solid. In certain
embodiments, a
solid compound(s) of Formula [X] is amorphous (lacks long-range order at the
molecular
level). In certain embodiments, a solid compound(s) of Formula [X] is
crystalline. The term
'crystalline' refers to a solid phase in which the material has a regular
ordered internal
structure at the molecular level and gives a distinctive X-ray diffraction
pattern with defined
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peaks. Compounds of the present invention may exist as polymorphs. As used
herein
"polymorph" refers to crystalline forms having the same chemical composition
but different
spatial arrangements of the molecules, and/or ions forming the crystal. Co-
crystals are
typically defined as crystalline complexes of neutral molecular constituents
that are bound
together through non-covalent interactions, but could also be a complex of a
neutral molecule
with a salt. Co-crystals may be prepared by melt crystallization, by
recrystallization from
solvents, or by physically grinding the components together; see 0. Almarsson
and M. J.
Zaworotko, Chem. Commun. 2004, 17, 1889-1896. For a general review of multi-
component
complexes, see J. K. Haleblian, J. Pharm. Sci. 1975, 64, 1269-1288. Reference
to a
.. compound of the Formula [X] herein is understood to include reference to
amorphous/crystal/polymorph/co-crystal/clathrate form thereof, unless
specified otherwise or
otherwise clear from context. A polymorph(s) of a compound(s), or a polymorph
of a
salt/solvate/hydrate/prodrug of a compound(s), of Formula (I), (II), (III),
(IV), (V), (VI) or
(VII), or a polymorph(s) of another compound(s) that reduces F IF ATP
hydrolysis, is
.. componentry to this invention, as is its incorporation, optionally of a
therapeutically effective
amount, into a pharmaceutically acceptable composition, optionally with a
further
polymorph(s) and/or compound(s) of this invention, optionally with an FDA
and/or EMA
approved therapeutic(s). An invention embodiment is the use of a polymorph(s)
of this
invention for the manufacture of a medicament for the treatment, amelioration,
prevention or
combating of a disease or disorder, optionally one or more
diseases/disorders/conditions
referred to herein. An invention embodiment is the use of one or more of these
polymorph(s)/composition(s) for use in a method of treatment of the human or
animal body
by therapy, optionally to treat/ameliorate/prevent/combat one or more
diseases/disorders/conditions referred to herein, optionally cancer. In some
embodiments, the
subject is further administered with one or more compounds or compositions
approved for
human use, optionally for anti-cancer use, by the United States Food and Drug
Administration (FDA) and/or European Medicines Agency (EMA), optionally in the
same
pharmaceutical composition. The present invention provides pharmaceutically
acceptable
crystalline forms. An aspect of the invention is a pharmaceutical composition
comprising at
least one polymorph of a compound as described herein and a pharmaceutically-
acceptable
carrier or diluent.
In one embodiment, a compound of the present invention is in substantially
pure form. The
term "substantially pure", as used herein, means a compound having a purity
greater than
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about 90% including greater than 90, 91, 92, 93, 94, 95, 96, 97, 98, and 99
weight %, and
also including equal to about 100 weight % of the compound, based on the
weight of the
compound. The remaining material comprises other form(s) of the compound,
and/or reaction
impurities and/or processing impurities arising from its preparation. For
example, a
.. crystalline form of a compound may be deemed substantially pure in that it
has a purity
greater than 90 weight %, as measured by means that are at this time known and
generally
accepted in the art, where the remaining less than 10 weight % of material
comprises other
form(s) of the compound and/or reaction impurities and/or processing
impurities.
.. Samples of the crystalline forms may be provided with substantially pure
phase homogeneity,
indicating the presence of a dominant amount of a single crystalline form and
optionally
minor amounts of one or more other crystalline forms. The presence of more
than one
crystalline form in a sample may be determined by techniques such as powder X-
ray
diffraction (PXRD) or solid state nuclear magnetic resonance spectroscopy
(SSNMR). For
example, the presence of extra peaks in the comparison of an experimentally
measured
PXRD pattern with a simulated PXRD pattern may indicate more than one
crystalline form in
the sample. The simulated PXRD may be calculated from single crystal X-ray
data. see
Smith, D. K., A FORTRAN Program for Calculating X- Ray Powder Diffraction
Patterns,"
Lawrence Radiation Laboratory, Livermore, Calif, UCRL-7196, April 1963.
Preferably, the
crystalline form has substantially pure phase homogeneity as indicated by less
than 10%,
preferably less than 5%, and more preferably less than 2% of the total peak
area in the
experimentally measured PXRD pattern arising from the extra peaks that are
absent from the
simulated XRPD pattern. Most preferred is a crystalline form having
substantially pure phase
homogeneity with less than 1% of the total peak area in the experimentally
measured PXRD
pattern arising from the extra peaks that are absent from the simulated PXRD
pattern.
The crystalline forms may be prepared by a variety of methods, including for
example,
crystallization or recrystallization from a suitable solvent, sublimation,
growth from a melt,
solid state transformation from another phase, crystallization from a
supercritical fluid, and
jet spraying. Techniques for crystallization or recrystallization of
crystalline forms from a
solvent mixture include, for example, evaporation of the solvent, decreasing
the temperature
of the solvent mixture, crystal seeding a supersaturated solvent mixture of
the molecule
and/or salt, freeze drying the solvent mixture, and addition of antisolvents
(countersolvents)
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to the solvent mixture. High throughput crystallization techniques may be
employed to
prepare crystalline forms including polymorphs.
Crystals of drugs, including polymorphs, methods of preparation, and
characterization of
.. drug crystals are discussed in Solid- State Chemistry of Drugs, S. R. Bym,
R. R. Pfeiffer, and
J. G. Stowell, 2"d Edition, SSCI, West Lafayette, Ind., 1999.
For crystallization techniques that employ solvent, the choice of solvent or
solvents is
typically dependent upon one or more factors, such as solubility of the
compound,
.. crystallization technique, and vapor pressure of the solvent. Combinations
of solvents may be
employed, for example, the compound may be solubilized into a first solvent to
afford a
solution, followed by the addition of an antisolvent to decrease the
solubility of the
compound in the solution and to afford the formation of crystals. An
antisolvent is a solvent
in which the compound has low solubility. Suitable solvents for preparing
crystals include
.. polar and nonpolar solvents.
In one method to prepare crystals, the compound of the present invention is
suspended and/or
stirred in a suitable solvent to afford a slurry, which may be heated to
promote dissolution.
The term "slurry", as used herein, means a saturated solution of the compound
and a solvent
at a given temperature. Suitable solvents in this regard include, for example,
polar aprotic
solvents, and polar protic solvents, and nonpolar solvents, and mixtures of
two or more of
these.
Suitable polar aprotic solvents include, for example, dicholomethane (CH 2CI 2
or DCM),
tetrahydrofuran (THF), acetone, methyl ethyl ketone (MEK), dimethylformamide
(DMF),
dimethylacetamide (DMAC), 1,3-dimethy1-3,4,5,6-tetrahydro-2(1H)-pyrimidinone
(DMPU),
1,3-dimethy1-2-imidazolidinone (DM I), N-methylpyrrolidinone (NMP), formamide,
N-
methylacetamide, N-methylformamide, acetonitrile (ACN or MeCN),
dimethylsulfoxide
(DMSO), propionitrile, ethyl formate, methyl acetate (Me0Ac), ethyl acetate
(Et0Ac),
isopropyl acetate (Ip0Ac), butyl acetate (BuOAc), t-butyl acetate,
hexachloroacetone,
dioxane, sulfolane, N,N-dimethylpropionamide, nitromethane, nitrobenzene and
hexamethylphosphoramide.
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Suitable polar protic solvents include, for example, alcohols and glycols,
such as H 20,
methanol, ethanol, 1-propanol, 2-propanol, isopropanol (IPA), 1-butanol (1-
BuOH), 2-
butanol (2-BuOH), i-butyl alcohol, t-butyl alcohol, 2-nitroethanol, 2-
fluoroethanol, 2,2,2-
trifluoroethanol, ethylene glycol, 2-methoxyethanol, 2-ethoxyethanol,
diethylene glycol, 1-,
2-, or 3-pentanol, neo-pentyl alcohol, t-pentyl alcohol, diethylene glycol
monomethyl ether,
diethylene glycol monoethyl ether, cyclohexanol, benzyl alcohol, phenol,
glycerol and methyl
t-butyl ether (MTBE).
Preferred solvents include, for example, acetone, H 20, CH 2C12, methanol,
ethanol, MEK,
IPA, and Et0Ac. Other solvents suitable for the preparation of slurries, in
addition to those
exemplified above, would be apparent to one skilled in the art, based on the
present
disclosure.
Seed crystals may be added to any crystallization mixture to promote
crystallization. As will
be clear to the skilled artisan, seeding is used as a means of controlling
growth of a particular
crystalline form or as a means of controlling the particle size distribution
of the crystalline
product. Accordingly, calculation of the amount of seeds needed depends on the
size of the
seed available and the desired size of an average product particle as
described, for example,
in "Programmed cooling of batch crystallizers," J. W. Mullin and J. Nyvlt,
Chemical
Engineering Science, 1971, 26, 369-377. In general, seeds of small size are
needed to
effectively control the growth of crystals in the batch. Seeds of small size
may be generated
by sieving, milling, or micronizing of larger crystals, or by micro-
crystallization of solutions.
Care should be taken that milling or micronizing of crystals does not result
in any change in
crystallinity of the desired crystal form or form conversions (i.e. change to
amorphous or to
another polymorph).
A cooled mixture may be filtered under vacuum, and the isolated solids may be
washed with
a suitable solvent, such as cold recrystallization solvent, and dried under a
nitrogen purge to
afford the desired crystalline form. The isolated solids may be analyzed by a
suitable
spectroscopic or analytical technique, such as SSNMR, DSC, PXRD, or the like,
to assure
formation of the preferred crystalline form of the product. The resulting
crystalline form is
typically produced in an amount of greater than about 70 weight % isolated
yield, but
preferably greater than 90 weight % based on the weight of the compound
originally
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employed in the crystallization procedure. The product may be comilled or
passed through a
mesh screen to delump the product, if necessary.
Crystalline forms may be prepared directly from the reaction medium of the
final process step
for preparing the compound of the present invention. This may be achieved, for
example, by
employing in the final process step a solvent or mixture of solvents from
which the
compound may be crystallized. Alternatively, crystalline forms may be obtained
by
distillation or solvent addition techniques. Suitable solvents for this
purpose include any of
those solvents described herein, including protic polar solvents such as
alcohols, and aprotic
polar solvents such as ketones.
By way of general guidance, the reaction mixture may be filtered to remove any
undesired
impurities, inorganic salts, and the like, followed by washing with reaction
or crystallization
solvent. The resulting solution may be concentrated to remove excess solvent
or gaseous
constituents. If distillation is employed, the ultimate amount of distillate
collected may vary,
depending on process factors including, for example, vessel size, stirring
capability, and the
like, by way of general guidance, the reaction solution may be distilled to
about 1/10 the
original volume before solvent replacement is carried out. The reaction may be
sampled and
assayed to determine the extent of the reaction and the wt % product in
accordance with
standard process techniques. If desired, additional reaction solvent may be
added or removed
to optimize reaction concentration. Preferably, the final concentration is
adjusted to about 50
wt % at which point a slurry typically results.
It may be preferable to add solvents directly to the reaction vessel without
distilling the
reaction mixture. Preferred solvents for this purpose are those which may
ultimately
participate in the crystalline lattice as discussed above in connection with
solvent exchange.
Although the final concentration may vary depending on desired purity,
recovery and the
like, the final concentration of the in solution is preferably about 4% to
about 7%. The
reaction mixture may be stirred following solvent addition and simultaneously
warmed. By
way of illustration, the reaction mixture may be stirred for about 1 hour
while warming to
about 70 C. The reaction is preferably filtered hot and washed with either
the reaction
solvent, the solvent added or a combination thereof. Seed crystals may be
added to any
crystallization solution to initiate crystallization.
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The various forms described herein may be distinguishable from one another
through the use
of various analytical techniques known to one of ordinary skill in the art.
Such techniques
include, but are not limited to, solid state nuclear magnetic resonance
(SSNMR)
spectroscopy, X-ray powder diffraction (PXRD), differential scanning
calorimetry (DSC),
and/or thermogravimetric analysis (TGA).
The crystalline forms of the compound of the present invention may be
formulated into
pharmaceutical compositions and/or employed in therapeutic and/or prophylactic
methods.
These methods include, but are not limited to, the administration of the
crystalline compound,
alone or in combination with one or more other pharmaceutically active agents,
including
agents that may be useful in the treatment of the disorders mentioned herein.
The methods
preferably comprise administering to a patient a pharmaceutically effective
amount of the
novel crystals of the present invention, preferably in combination with one or
more
pharmaceutically acceptable carriers and/or excipients. The relative
proportions of active
ingredient and carrier and/or excipient may be determined, for example, by the
solubility and
chemical nature of the materials, chosen route of administration and standard
pharmaceutical
practice.
The crystalline forms of the compound may be administered to a patient in such
oral dosage
forms as tablets, capsules (each of which includes sustained release or timed
release
formulations), pills, powders, granules, elixirs, tinctures, suspensions,
syrups, and emulsions.
They may also be administered in intravenous (bolus or infusion),
intraperitoneal,
subcutaneous, or intramuscular form, all using dosage forms well known to
those of ordinary
skill in the pharmaceutical arts. They may be administered alone, but
generally will be
administered with a pharmaceutical carrier selected on the basis of the chosen
route of
administration and standard pharmaceutical practice.
The dosage regimen for the crystalline forms of the compound will, of course,
vary
depending upon known factors, such as the pharmacodynamic characteristics of
the particular
.. agent and its mode and route of administration; the species, age, sex,
health, medical
condition, and weight of the recipient; the nature and extent of the symptoms;
the kind of
concurrent treatment; the frequency of treatment; the route of administration,
the renal and
hepatic function of the patient, and the effect desired. A physician or
veterinarian can
determine and prescribe the effective amount of the drug required to prevent,
counter, or
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arrest the disease/disorder/condition. Obviously, several unit dosage forms
may be
administered at about the same time. The dosage of the crystalline form of the
compound that
will be most suitable for prophylaxis or treatment may vary with the form of
administration,
the particular crystalline form of the compound chosen and the physiological
characteristics
of the particular patient under treatment. Broadly, small dosages may be used
initially and, if
necessary, increased by small increments until the desired effect under the
circumstances is
reached.
By way of general guidance, in the adult, suitable doses may range from about
0.001 to about
1000 mg/Kg body weight, and all combinations and subcombinations of ranges and
specific
doses therein. Preferred doses may be from about 0.01 to about 100 mg/kg body
weight per
day by inhalation, preferably 0.1 to 70, more preferably 0.5 to 20 mg/Kg body
weight per day
by oral administration, and from about 0.01 to about 50, preferably 0.01 to 10
mg/Kg body
weight per day by intravenous administration. In each particular case, the
doses may be
determined in accordance with the factors distinctive to the subject to be
treated, such as age,
weight, general state of health and other characteristics which can influence
the efficacy of
the medicinal product. The crystalline forms of the compound may be
administered in a
single daily dose, or the total daily dosage may be administered in divided
doses of two,
three, or four times daily.
For oral administration in solid form such as a tablet or capsule, the
crystalline forms of the
compound can be combined with a non-toxic, pharmaceutically acceptable inert
carrier, such
as lactose, starch, sucrose, glucose, methylcellulose, magnesium stearate,
dicalcium
phosphate, calcium sulfate, mannitol, sorbitol and the like. Preferably, in
addition to the
active ingredient, solid dosage forms may contain a number of additional
ingredients referred
to herein as "excipients". These excipients include among others diluents,
binders, lubricants,
glidants and disintegrants. Coloring agents may also be incorporated.
"Diluents", as used
herein, are agents which impart bulk to the formulation to make a tablet a
practical size for
compression. Examples of diluents are lactose and cellulose. "Binders", as
used herein, are
agents used to impart cohesive qualities to the powered material to help
ensure the tablet will
remain intact after compression, as well as improving the free-flowing
qualities of the
powder. Examples of typical binders are lactose, starch and various sugars.
"Lubricants", as
used herein, have several functions including preventing the adhesion of the
tablets to the
compression equipment and improving the flow of the granulation prior to
compression or
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encapsulation. Lubricants are in most cases hydrophobic materials. Excessive
use of
lubricants is undesired, however, as it may result in a formulation with
reduced disintegration
and/or delayed dissolution of the drug substance. "Glidants", as used herein,
refer to
substances which may improve the flow characteristics of the granulation
material. Examples
of glidants include talc and colloidal silicon dioxide. "Disintegrants", as
used herein, are
substances or a mixture of substances added to a formulation to facilitate the
breakup or
disintegration of the solid dosage form after administration. Materials that
may serve as
disintegrants include starches, clays, celluloses, algins, gums and cross-
linked polymers. A
group of disintegrants referred to as "super-disintegrants" generally are used
at a low level in
the solid dosage form, typically 1% to 10% by weight relative to the total
weight of the
dosage unit. Croscarmelose, crospovidone and sodium starch glycolate represent
examples of
a cross-linked cellulose, a cross-linked polymer and a cross-linked starch,
respectively.
Sodium starch glycolate swells seven- to twelve-fold in less than 30 seconds
effectively
disintegrating the granulations that contain it.
The disintegrant preferably used in the present invention is selected from the
group
comprising modified starches, croscarmallose sodium, carboxymethylcellulose
calcium and
crospovidone. A more preferred disintegrant in the present invention is a
modified starch
such as sodium starch glycolate.
Preferred carriers include capsules or compressed tablets which contain the
solid
pharmaceutical dosage forms described herein. Preferred capsule or compressed
tablet forms
generally comprise a therapeutically effective amount of the crystalline forms
of the
compound and one or more disintegrants in an amount greater than about 10% by
weight
relative to the total weight of the contents of the capsule or the total
weight of the tablet.
Preferred capsule formulations may contain the crystalline forms of the
compound in an
amount from about 5 to about 1000 mg per capsule. Preferred compressed tablet
formulations
contain the crystalline forms of the compound in an amount from about 5 mg to
about 800
mg per tablet. More preferred formulations contain about 50 to about 200 mg
per capsule or
compressed tablet. Preferably, the capsule or compressed tablet pharmaceutical
dosage form
comprises a therapeutically effective amount of the crystalline forms; a
surfactant; a
disintegrant; a binder; a lubricant; and optionally additional
pharmaceutically acceptable
excipients such as diluents, glidants and the like; wherein the disintegrant
is selected from
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modified starches; croscarmallose sodium, carboxymethylcellulose calcium and
crospovidone.
For oral administration in liquid form, the crystalline forms of the compound
can be
combined with any oral, non-toxic pharmaceutically acceptable inert carrier
such as ethanol,
glycerol, water and the like. The liquid composition may contain a sweetening
agent which to
make the compositions more palatable. The sweetening agent can be selected
from a sugar
such as sucrose, mannitol, sorbitol, xylitol, lactose, etc. or a sugar
substitute such as
cyclamate, saccaharin, aspartame, etc. If sugar substitutes are selected as
the sweetening
agent the amount employed in the compositions of the invention will be
substantially less
than if sugars are employed. Taking this into account, the amount of
sweetening agent may
range from about 0.1 to about 50% by weight, and all combinations and
subcombinations of
ranges and specific amounts therein. Preferred amounts range from about 0.5 to
about 30%
by weight. The more preferred sweetening agents are the sugars and
particularly sucrose. The
particle size of the powdered sucrose used has been found to have a
significant influence in
the physical appearance of the finished composition and its ultimate
acceptance for taste. The
preferred particle size of the sucrose component when used is in the range of
from 200 to less
than 325 mesh US Standard Screen, and all combinations and subcombinations of
ranges and
specific particle sizes therein.
Sterile injectable solutions may be prepared by incorporating the crystalline
forms of the
compound in the required amounts, in the appropriate solvent, with various of
the other
ingredients enumerated herein, as required, followed by filtered
sterilization. Generally,
dispersions may be prepared by incorporating the sterilized active ingredient
into a sterile
vehicle which contains the dispersion medium and any other required
ingredients. In the case
of sterile powders for the preparation of sterile injectable solutions, the
preferred methods of
preparation may include vacuum drying and the freeze drying technique which
may yield a
powder of the active ingredient, plus any additional desired ingredient from
the previously
sterile-filtered solution thereof.
As would be apparent to a person of ordinary skill in the art, once armed with
the teachings
of the present disclosure, when dissolved, a crystalline compound loses its
crystalline
structure, and is therefore considered to be a solution of the compound. All
forms of the
present invention, however, may be used for the preparation of liquid
formulations in which
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the compound may be, for example, dissolved or suspended. In addition, the
crystalline forms
of the compound may be incorporated into solid formulations.
The liquid compositions may also contain other components routinely utilized
in formulating
pharmaceutical compositions. One example of such components is lecithin. Its
use in
compositions of the invention as an emulsifying agent in the range of from
0.05 to 1% by
weight, and all combinations and subcombinations of ranges and specific
amounts therein.
More preferably, emulsifying agents may be employed in an amount of from about
0.1 to
about 0.5% by weight. Other examples of components that may be used are
antimicrobial
preservatives, such as benzoic acid or parabens; suspending agents, such as
colloidal silicon
dioxide; antioxidants; topical oral anesthetics; flavoring agents; and
colorants.
The selection of such optional components and their level of use in the
compositions of the
invention is within the level of skill in the art and will be even better
appreciated from the
working examples provided hereinafter.
The crystalline forms of the compound may also be coupled with soluble
polymers as
targetable drug carriers. Such polymers can include polyvinylpyrrolidine pyran
copolymer,
polyhydroxypropylmethacrylamide-phenol, polyhydroxyethyl-aspartamidephenol or
polyethylene oxide-polylysine substituted with palmitolyl residues.
Furthermore, the
crystalline compound may be coupled to a class of biodegradable polymers
useful in
achieving controlled release of a drug, for example, polylactic acid,
polyglycolic acid,
copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone,
polyhydroxy
butyric acid, polyorthoesters, polyacetals, polydihydropyrans,
polycyanoacrylates and
crosslinked or amphipathic block copolymers of hydrogels.
Gelatin capsules of the crystalline forms of the compound may contain the
crystalline
compound and the liquid or solid compositions described herein. Gelatin
capsules may also
contain powdered carriers such as lactose, starch, cellulose derivatives,
magnesium stearate,
stearic acid and the like. Similar diluents can be used to make compressed
tablets. Both
tablets and capsules can be manufactured as sustained release products to
provide for
continuous release of medication over a period of hours. Tablets can be sugar
coated or film
coated to mask any unpleasant taste and to protect the tablet from the
atmosphere or enteric
coated for selective disintegration in the gastrointestinal track.
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In general, water, a suitable oil, saline, aqueous dextrose (glucose), and
related sugar
solutions and glycols, such as propylene glycol or polyethylene glycols are
suitable carriers
for parenteral solutions. Solutions for parenteral solutions are prepared by
dissolving the
crystalline compound in the carrier and, if necessary, adding buffering
substances. Anti-
oxidizing agents such as sodium bisulfite, sodium sulfite, or ascorbic acid
either alone or
combined, are suitable stabilizing agents. Citric acid and its salts and
sodium EDTA may also
be employed. Parenteral solutions may also contain preservatives, such as
benzalkonium
chloride, methyl- or propyl-paraben and chlorobutanol.
Suitable pharmaceutical carriers are described in Remington's Pharmaceutical
Sciences,
Mack Publishing Co., the disclosures of which are hereby incorporated herein
by reference,
in their entireties.
The crystalline forms of the compound of the present invention and
pharmaceutical
composition thereof may be administered to treat/ameliorate/prevent/combat
cancer in a
subject. Accordingly, the present invention provides methods for the treatment
and/or
prevention of cancer in mammals. Crystalline forms of the compound may be used
alone or
in combination with other anti-cancer agent(s). The preferred crystalline form
of the
compound may serve as component (a) of this invention and can independently be
in any
dosage form, such as those described above, and can also be administered in
various
combinations, as described above. In the following description component (b)
is to be
understood to represent one or more agents as described herein suitable for
combination
therapy. Accordingly, components (a) and (b) of the present invention may be
formulated
together, in a single dosage unit (that is, combined together in one capsule,
tablet, powder, or
liquid, etc.) as a combination product. When component (a) and (b) are not
formulated
together in a single dosage unit, the component (a) may be administered at the
same time as
component (b) or in any order; for example component (a) of this invention may
be
administered first, followed by administration of component (b), or they may
be administered
in the reverse order. If component (b) contains more than one agent, these
agents may be
administered together or in any order. When not administered at the same time,
preferably the
administration of component (a) and (b) occurs less than about one hour apart.
Preferably, the
route of administration of component (a) and (b) is oral. Although it may be
preferable that
component (a) and component (b) both be administered by the same route (that
is, for
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CA 3050553 2019-07-25
example, both orally) or dosage form, if desired, they may each be
administered by different
routes (that is, for example, one component of the combination product may be
administered
orally, and another component may be administered intravenously) or dosage
forms.
Co-administration
A compound(s) of Formula [X] can be combined with one or more other
pharmacologically
active compounds ("second active agents") in methods and compositions provided
herein. As
used herein, the term "co-administration" refers to the administration of at
least two
agent(s)/therapies, one or more of which is a compound or composition of the
present
invention. In some embodiments, the co-administration of two or more
agents/therapies is
concurrent. In other embodiments, a first agent/therapy is administered prior
to a second
agent/therapy. Those of skill in the art understand that the formulations
and/or routes of
administration of the various agents/therapies used may vary. The appropriate
dosage for co-
administration can be readily determined by one skilled in the art. In some
embodiments,
when agents/therapies are co-administered, the respective agents/therapies are
administered at
lower dosages than appropriate for their administration alone. Thus, co-
administration is
especially desirable in embodiments where the co-administration of the
agents/therapies
lowers the requisite dosage of a known potentially harmful (e.g., toxic)
agent(s). The use of
multiple compounds to treat an indication can increase the beneficial effects
while reducing
the presence of side effects.
The compounds of the present disclosure may be administered individually, in
combination
with each other, and/or in combination with other pharmaceuticals/treatments
useful for
treating the disease or condition of interest. It is understood that the
component medications
of a combination therapy may be administered to the patient simultaneously or
at different
times, and may be administered on the same or different dosing schedules, as
appropriate.
When administered in combination, each component may be administered at the
same time or
sequentially in any order at different points in time. Thus, each component
may be
administered separately but sufficiently closely in time so as to provide the
desired
therapeutic effect, in a way that the therapeutical effects of the first
administered one is not
entirely dissipated when the subsequent is administered. The dosage of each
component
medication need not be the same and are expected to be different in most
cases. Furthermore,
the routes of administration may be the same or different. In one embodiment,
the component
medications are coformulated for convenience (e.g. in the same injectable or
ingestible
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composition). In an alternative embodiment, the component medications are
packaged/distributed/sold/promoted/advertised together.
Particularly when provided as a single dosage unit, the potential exists for a
chemical
interaction between the combined active ingredients. For this reason, when the
compound of
the present invention and a second therapeutic agent are combined in a single
dosage unit
they are formulated such that although the active ingredients are combined in
a single dosage
unit, the physical contact between the active ingredients is minimized (that
is, reduced). For
example, one active ingredient may be enteric coated. By enteric coating one
of the active
ingredients, it is possible not only to minimize the contact between the
combined active
ingredients, but also, it is possible to control the release of one of these
components in the
gastrointestinal tract such that one of these components is not released in
the stomach but
rather is released in the intestines. One of the active ingredients may also
be coated with a
material that affects a sustained-release throughout the gastrointestinal
tract and also serves to
minimize physical contact between the combined active ingredients.
Furthermore, the
sustained-released component can be additionally enteric coated such that the
release of this
component occurs only in the intestine. Still another approach would involve
the formulation
of a combination product in which the one component is coated with a sustained
and/or
enteric release polymer, and the other component is also coated with a polymer
such as a low
viscosity grade of hydroxypropyl methylcellulose (HPMC) or other appropriate
materials as
known in the art, in order to further separate the active components. The
polymer coating
serves to form an additional barrier to interaction with the other component.
These as well as other ways of minimizing contact between the components of
combination
products of the present invention, whether administered in a single dosage
form or
administered in separate forms but at the same time by the same manner, will
be readily
apparent to those skilled in the art, once armed with the present disclosure.
For cancer indications, additional such agents include, but are not limited
to, kinase
inhibitors, such as EGFR inhibitors (e.g., erlotinib, gefitinib), Raf
inhibitors (e.g.,
vemurafenib), VEGFR inhibitors (e.g., sunitinib), ALK inhibitors (e.g.,
crizotinib) standard
chemotherapy agents such as alkylating agents, antimetabolites, anti-tumor
antibiotics,
topoisomerase inhibitors, platinum drugs, mitotic inhibitors, antibodies,
hormone therapies,
or corticosteroids. For pain indications, suitable combination agents include
anti-
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CA 3050553 2019-07-25
inflammatories such as NSAIDs. The pharmaceutical compositions of the
invention may
additional comprise one or more of such active agents, and methods of
treatment may
additionally comprise administering an effective amount of one or more of such
active
agents.
Further additional active ingredients include other therapeutics or agents
that mitigate adverse
effects of therapies for the intended disease targets. Such combinations may
serve to increase
efficacy, ameliorate other disease symptoms, decrease one or more side
effects, or decrease
the required dose of an inventive compound. The additional active ingredients
may be
administered in a separate pharmaceutical composition from a compound of the
present
invention or may be included with a compound of the present invention in a
single
pharmaceutical composition. The additional active ingredients may be
administered
simultaneously with, prior to, or after administration of a compound of the
present invention.
Provided herein are methods of treating patients who have been previously
treated for cancer
but are non-responsive to standard therapies, as well as those who have not
previously been
treated. The invention further encompasses methods of treating patients who
have undergone
surgery in an attempt to treat the disease or condition at issue, as well as
those who have not.
Because patients with cancer have heterogeneous clinical manifestations and
varying clinical
outcomes, the treatment given to a patient may vary, depending on his/her
prognosis. The
skilled clinician will be able to readily determine without undue
experimentation specific
secondary agents, types of surgery, and types of non-drug based standard
therapy to
administer to a patient administered with a compound(s) of Formula [X], which
can be
effectively used to treat an individual subject with cancer.
A compound(s) provided herein, e.g. a compound(s) of Formula [X], or an
enantiomer or a
mixture of enantiomers thereof, or a pharmaceutically acceptable salt,
solvate, hydrate, co-
crystal, clathrate, or polymorph thereof, can be combined with one or more
other
pharmacologically active compounds ("second active agents") in methods and
compositions
provided herein. It is believed that certain combinations work synergistically
in the treatment
of particular types of cancer. The compound(s) of Formula [X] provided herein
can also work
to alleviate adverse effects associated with certain second active agents. One
or more second
active ingredients or agents can be used in the methods and compositions
provided herein
with the compound(s) of Formula [X] provided herein. Administration of a
compound(s) of
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CA 3050553 2019-07-25
Formula [X] and one or more second active agents to a subject can occur
simultaneously or
sequentially by the same or different routes of administration. Second active
agents can be
large molecules (e.g., proteins) or small molecules (e.g., synthetic
inorganic, organometallic,
or organic molecules). Examples of small molecule second active agents
include, but are not
.. limited to, anti-cancer agents, anti-neoplastic agents, anti-proliferation
agents,
chemotherapeutics, antibiotics, painkillers, anti-depressants, anti-anxiety
drugs, anti-nausea
drugs, antihistamines, immunosuppressive agents, steroids, anti-angiogenic
agents, anti-
fibrotic agents, anti-inflammatory agents, immune modulating agents,
immunotherapeutic
agents, therapeutic antibodies, radiotherapeutic agents, (i) antimetabolites;
(ii) DNA-
.. fragmenting agents, (iii) DNA-crosslinking agents, (iv) intercalating
agents (v) protein
synthesis inhibitors, (vi) topoisomerase I poisons, such as camptothecin or
topotecan; (vii)
topoisomerase II poisons, (viii) microtubule-directed agents, (ix) kinase
inhibitors (x)
miscellaneous investigational agents (xi) hormones and (xii) hormone
antagonists.
Examples of large molecule active agents include, but are not limited to,
immunotherapies, immuno-oncology agents, hematopoietic growth factors,
cytokines,
monoclonal and polyclonal antibodies e.g. antagonistic antibodies to one or
more of PD-L1,
CD27, CD137, CD25, CD40, OX4OL, IDO, GITR, LAG-3, CTLA-4. In certain
embodiments, large molecule active agents are biological molecules, such as
naturally
occurring or artificially made proteins. Proteins that are particularly useful
in this disclosure
.. include proteins that stimulate the survival and/or proliferation of
hematopoietic precursor
cells and immunologically active poietic cells in vitro or in vivo. Others
stimulate the division
and differentiation of committed erythroid progenitors in cells in vitro or in
vivo. Particular
proteins include, but are not limited to: interleukins, such as IL-2
(including recombinant IL-
II ("rIL2") and canarypox 1L-2), IL-10, IL-12, and IL-18; interferons, such as
interferon alfa-
2a, interferon alfa-2b, interferon alfa-nl, interferon alfa-n3, interferon
beta-I a, and interferon
gamma-I b; GM-CF and GM-CSF; and EPO.
Particular proteins that can be used in the methods and compositions of the
disclosure
include, but are not limited to: filgrastim, which is sold in the United
States under the trade
name NEUPOGEN (Amgen, Thousand Oaks, Calif.); sargramostim, which is sold in
the
United States under the trade name LEUKINE (Immunex, Seattle, Wash.); and
recombinant
EPO, which is sold in the United States under the trade name EPGEN (Amgen,
Thousand
Oaks, Calif.).
Inhibitors of ActRII receptors or activin-ActRII inhibitors may be used in the
methods
and compositions provided herein. ActRII receptors include ActRIIA inhibitors
and ActRIIB
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CA 3050553 2019-07-25
inhibitors. Inhibitors of ActRII receptors can be polypeptides comprising
activin-binding
domains of ActRII. In certain embodiments, the activin-binding domain
comprising
polypeptides are linked to an Fc portion of an antibody (i.e., a conjugate
comprising an
activin-binding domain comprising polypeptide of an ActRII receptor and an Fe
portion of an
antibody is generated). In certain embodiments, the activin-binding domain is
linked to an Fe
portion of an antibody via a linker, e.g., a peptide linker. Examples of such
non-antibody
proteins selected for activin or ActRIIA binding and methods for design and
selection of the
same are found in WO/2002/088171, WO/2006/055689, WO/2002/032925,
WO/2005/037989, US 2003/0133939, and US 2005/0238646, each of which is
incorporated
herein by reference in its entirety.
Recombinant and mutated forms of GM-CSF can be prepared as described in U.S.
Pat.
Nos. 5,391,485; 5,393,870; and 5,229,496; the disclosure of each of which is
incorporated
herein by reference in its entirety. Recombinant and mutated forms of G-CSF
can be prepared
as described in U.S. Pat. Nos. 4,810,643; 4,999,291; 5,528,823; and 5,580,755;
the disclosure
of each of which is incorporated herein by reference in its entirety.
This disclosure encompasses the use of native, naturally occurring, and
recombinant
proteins. The disclosure further encompasses mutants and derivatives (e.g.,
modified forms)
of naturally occurring proteins that exhibit, in vivo, at least some of the
pharmacological
activity of the proteins upon which they are based. Examples of mutants
include, but are not
limited to, proteins that have one or more amino acid residues that differ
from the
corresponding residues in the naturally occurring forms of the proteins. Also
encompassed by
the term "mutants" are proteins that lack carbohydrate moieties normally
present in their
naturally occurring forms (e.g., nonglycosylated forms). Examples of
derivatives include, but
are not limited to, pegylated derivatives and fusion proteins, such as
proteins formed by
fusing IgG1 or IgG3 to the protein or active portion of the protein of
interest. See, e.g.,
Penichet, M. L. and Morrison, S. L., J. Immunol. Methods 248:91-101 (2001).
One or more antibodies (or antibody fragments) and/or antibody-drug conjugates
can be
used in combination(s) with a compound(s) of Formula [X] including, but not
limited to,
monoclonal, polyclonal, multi-specific, murine, chimeric, humanized and human
antibodies.
Examples of antibodies include, but are not limited to, immune checkpoint
inhibitors,
including an antibody that targets one or more of PD-1 (CD279), PDL-1 (CD274),
PDL-2,
CTLA4, TIM3, 0X40, GITR, LAG3, CD137, 4-1BB, B7-H3, B7-H4, KIR, PARP, CD27,
ICOS, TIGIT, BTLA, VISTA, A2AR, CD80, CD86, CD-47, GD2, CSF1R, or antibodies
that
target cancer expressed antigens, which may or not be also expressed by normal
cells, such
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CA 3050553 2019-07-25
as, to illustrate, (on B cells) CD20, CD22, CD52, CD23, ROR-1. Exemplary
antibodies
include, but are not limited to, pembrolizumab (Keytruda), nivolumab (Opdivo),
durvalumab,
avelumab, atezolizumab (Tecentriq), ipilimumab (Yervoy), pidilizumab, AMP-224,
AMP-
514, PDR001, cemiplimab, BMS-936559, CK-301, pexidartinib, PLX7486, ARRY-382,
JNJ-
40346527, BLZ945, emactuzumab, AMG820, IMC-CS4, MCS110, cabiralizumab,
abagovomab, adecatumumab, afutuzumab, alemtuzumab, altumomab, amatuximab,
anatumomab, arcitumomab, bavituximab, bectumomab, bevacizumab, bivatuzumab,
blinatumomab, brentuximab, cantuzumab, catumaxomab, cetuximab, citatuzumab,
cixutumumab, clivatuzumab, conatumumab, daratumumab, drozitumab, duligotumab,
dusigitumab, detumomab, dacetuzumab, dalotuzumab, ecromeximab, elotuzumab,
ensituximab, ertumaxomab, etaracizumab, farietuzumab, ficlatuzumab,
figitumumab,
flanvotumab, futuximab, ganitumab, gemtuzumab, girentuximab, glembatumumab,
ibritumomab, igovomab, imgatuzumab, indatuximab, inotuzumab, intetumumab,
ipilimumab,
iratumumab, labetuzumab, lexatumumab, lintuzumab, lorvotuzumab, lucatumumab,
mapatumumab, matuzumab, milatuzumab, minretumomab, mitumomab, moxetumomab,
narnatumab, naptumomab, necitumumab, nimotuzumab, nofetumomabn, ocaratuzumab,
ofatumumab, olaratumab, onartuzumab, oportuzumab, oregovomab, panitumumab,
parsatuzumab, patritumab, pemtumomab, pertuzumab, pintumomab, pritumumab,
racotumomab, radretumab, rilotumumab, rituximab, robatumumab, satumomab,
sibrotuzumab, siltuximab, simtuzumab, solitomab, tacatuzumab, taplitumomab,
tenatumomab, teprotumumab, tigatuzumab, tositumomab, trastuzumab, tucotuzumab,
ublituximab, veltuzumab, vorsetuzumab, votumumab, zalutumumab, CC49 and 3F8.
trastuzumab (HERCEPTINg), rituximab (RITUXANt), bevacizumab (AVASTINTm),
pertuzumab (OMNITARGTm), tositumomab (BEXXARO), edrecolomab (PANOREXg),
panitumumab and G250. The exemplified therapeutic antibodies may be further
labeled or
combined with a radioisotope particle, such as indium In-111, yttrium Y-90,
iodine 1-131
(radioimmunotherapy). A compound(s) of Formula [X], or
salt(s)/hydrate(s)/solvate(s)/prodrug(s) thereof, can be combined with or used
in combination
with one or more of cytokine(s) therapy, such as interferon(s) and/or
interleukin(s)
administration, oncolytic virus therapy or therapies, cancer vaccine(s)
administration,
antigen(s)/tumour antigen(s)/neoantigen(s) administration, CpG
oligodeoxynucleotide(s),
immunostimulant(s), dendritic cell therapy, adoptive cell transfer, CAR-T cell
therapy,
antibody therapy, anti-TNF-a antibody(ies), combination immunotherapy,
immunotherapy,
cancer immunotherapy, immuno-oncology therapy or any treatment(s), most
preferably FDA
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CA 3050553 2019-07-25
approved, that utilises/leverages a subject's immune system or an immune
system
component(s) or mimetic(s) to treat cancer in a subject, optionally in further
combination
with one or more other cancer treatment(s) including, but not limited to,
chemotherapy,
targeted agent(s), radiation etc.
Examples of anti-cancer agents include, but are not limited to: abraxane; ace-
11; acivicin;
aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin;
altretamine;
ambomycin; ametantrone acetate; amrubicin; amsacrine; anastrozole;
anthramycin;
asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat;
benzodepa;
bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin;
bleomycin sulfate;
brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone;
caracemide; carbetimer;
carboplatin; carmustine; carubicin hydrochloride; carzelesin; cedefingol;
celecoxib (COX-2
inhibitor); chlorambucil; cirolemycin; cisplatin; cladribine; crisnatol
mesylate;
cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicin
hydrochloride;
decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone;
docetaxel;
doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifene citrate;
dromostanolone
propionate; duazomycin; edatrexate; eflomithine hydrochloride; elsamitrucin;
enloplatin;
enpromate; epipropidine; epirubicin hydrochloride; erbulozole; esorubicin
hydrochloride;
estramustine; estramustine phosphate sodium; etanidazole; etoposide; etoposide
phosphate;
etoprine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine;
fludarabine
phosphate; fluorouracil; flurocitabine; fosquidone; fostriecin sodium;
gemcitabine;
gemcitabine hydrochloride; herceptin; hydroxyurea; idarubicin hydrochloride;
ifosfamide;
ilmofosine; iproplatin; irinotecan; irinotecan hydrochloride; lanreotide
acetate; lapatinib;
letrozole; leuprolide acetate; liarozole hydrochloride; lometrexol sodium;
lomustine;
losoxantrone hydrochloride; masoprocol; maytansine; mechlorethamine
hydrochloride;
megestrol acetate; melengestrol acetate; melphalan; menogaril; mercaptopurine;
methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide;
mitocarcin;
mitocromin; mitogillin; mitomalcin; mitomycin; mitosper; mitotane;
mitoxantrone
hydrochloride; mycophenolic acid; nocodazole; nogalamycin; ormaplatin;
oxisuran;
paclitaxel; pegaspargase; peliomycin; pentamustine; peplomycin sulfate;
perfosfamide;
pipobroman; piposulfan; piroxantrone hydrochloride; plicamycin; plomestane;
porfimer
sodium; porfiromycin; prednimustine; procarbazine hydrochloride; puromycin;
puromycin
hydrochloride; pyrazofurin; riboprine; romidepsin; safingol; safingol
hydrochloride;
semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermanium
hydrochloride;
spiromustine; spiroplatin; stem cell treatments such as PDA-001;
streptonigrin; streptozocin;
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sulofenur; talisomycin; tecogalan sodium; taxotere; tegafur; teloxantrone
hydrochloride;
temoporfin; teniposide; teroxirone; testolactone; thiamiprine; thioguanine;
thiotepa;
tiazofurin; tirapazamine; toremifene citrate; trestolone acetate; triciribine
phosphate;
trimetrexate; trimetrexate glucuronate; triptorelin; tubulozole hydrochloride;
uracil mustard;
uredepa; vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate;
vindesine; vindesine
sulfate; vinepidine sulfate; vinglycinate sulfate; vinleurosine sulfate;
vinorelbine tartrate;
vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin;
and zorubicin
hydrochloride.
Other anti-cancer drugs include, but are not limited to: 20-epi-1,25
dihydroxyvitamin D3;
5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol;
adozelesin; aldesleukin;
ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine;
aminolevulinic acid;
amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis
inhibitors;
antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein-
1;
antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston; antisense
oligonucleotides;
.. aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators;
apurinic acid; ara-
CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane; atrimustine;
axinastatin 1;
axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatin III
derivatives; balanol;
batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta
lactam
derivatives; beta-alethine; betaclamycin B; betulinic acid; b-FGF inhibitor;
bicalutamide;
bisantrene; bisaziridinylspermine; bisnafide; bistratene A; bizelesin;
breflate; bropirimine;
budotitane; buthionine sulfoximine; calcipotriol; calphostin C; camptothecin
derivatives;
capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRest M3;
CARN 700;
cartilage derived inhibitor; carzelesin; casein kinase inhibitors (ICOS);
castanospermine;
cecropin B; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost;
cis-porphyrin;
cladribine; clomifene analogues; clotrimazole; collismycin A; collismycin B;
combretastatin
A4; combretastatin analogue; conagenin; crambescidin 816; crisnatol;
cryptophycin 8;
cryptophycin A derivatives; curacin A; cyclopentanthraquinones; cycloplatam;
cypemycin;
cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; decitabine;
dehydrodidemnin B;
deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil;
diaziquone;
didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine; dihydrotaxol, 9-
;
dioxamycin; diphenyl spiromustine; docetaxel; docosanol; dolasetron;
doxifluridine;
doxorubicin; droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine;
edelfosine;
edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride;
estramustine analogue;
estrogen agonists; estrogen antagonists; etanidazole; etoposide phosphate;
exemestane;
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fadrozole; fazarabine; fenretinide; filgrastim; finasteride; flavopiridol;
flezelastine;
fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex;
formestane;
fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate; galocitabine;
ganirelix;
gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;
heregulin;
hexamethylene bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene;
idramantone;
ilmofosine; ilomastat; imatinib (e.g., GLEEVECO), imiquimod; immunostimulant
peptides;
insulin-like growth factor-1 receptor inhibitor; interferon agonists;
interferons; interleukins;
iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact; irsogladine;
isobengazole;
isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F; lamellarin-N
triacetate;
lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin;
letrozole; leukemia
inhibiting factor; leukocyte alpha interferon;
leuprolide+estrogen+progesterone; leuprorelin;
levamisole; liarozole; linear polyamine analogue; lipophilic disaccharide
peptide; lipophilic
platinum compounds; lissoclinamide 7; lobaplatin; lombricine; lometrexol;
lonidamine;
losoxantrone; loxoribine; lurtotecan; lutetium texaphyrin; lysofylline; lytic
peptides;
maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysin
inhibitors; matrix
metalloproteinase inhibitors; menogaril; merbarone; meterelin; methioninase;
metoclopramide; MIF inhibitor; mifepristone; miltefosine; mirimostim;
mitoguazone;
mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growth
factor-saporin;
mitoxantrone; mofarotene; molgramostim; Erbitux, human chorionic
gonadotrophin;
.. monophosphoryl lipid A+myobacterium cell wall sk; mopidamol; mustard
anticancer agent;
mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-
acetyldinaline; N-
substituted benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin;
naphterpin;
nartograstim; nedaplatin; nemorubicin; neridronic acid; nilutamide; nisamycin;
nitric oxide
modulators; nitroxide antioxidant; nitrullyn; oblimersen (GENASENSE0); 0 6-
benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;
ondansetron;
ondansetron; oracin; oral cytokine inducer; ormaplatin; osaterone;
oxaliplatin; oxaunomycin;
paclitaxel; paclitaxel analogues; paclitaxel derivatives; palauamine;
palmitoylrhizoxin;
pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine;
pegaspargase; peldesine;
pentosan polysulfate sodium; pentostatin; pentrozole; perflubron;
perfosfamide; perillyl
alcohol; phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil;
pilocarpine
hydrochloride; pirarubicin; piritrexim; placetin A; placetin B; plasminogen
activator
inhibitor; platinum complex; platinum compounds; platinum-triamine complex;
porfimer
sodium; porfiromycin; prednisone; propyl bis-acridone; prostaglandin J2;
proteasome
inhibitors; protein A-based immune modulator; protein kinase C inhibitor;
protein kinase C
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CA 3050553 2019-07-25
inhibitors, microalgal; protein tyrosine phosphatase inhibitors; purine
nucleoside
phosphorylase inhibitors; purpurins; pyrazoloacridine; pyridoxylated
hemoglobin
polyoxyethylene conjugate; raf antagonists; raltitrexed; ramosetron; ras
farnesyl protein
transferase inhibitors; ras inhibitors; ras-GAP inhibitor; retelliptine
demethylated; rhenium
Re 186 etidronate; rhizoxin; ribozymes; RII retinamide; rohitukine; romurtide;
roquinimex;
rubiginone B1; ruboxyl; safingol; saintopin; SarCNU; sarcophytol A;
sargramostim; Sdi 1
mimetics; semustine; senescence derived inhibitor 1; sense oligonucleotides;
signal
transduction inhibitors; sizofiran; sobuzoxane; sodium borocaptate; sodium
phenylacetate;
solverol; somatomedin binding protein; sonermin; sparfosic acid; spicamycin D;
spiromustine; splenopentin; spongistatin 1; squalamine; stipiamide;
stromelysin inhibitors;
sulfinosine; superactive vasoactive intestinal peptide antagonist; suradista;
suramin;
swainsonine; tallimustine; tamoxifen methiodide; tauromustine; tazarotene;
tecogalan
sodium; tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;
teniposide;
tetrachlorodecaoxide; tetrazomine; thaliblastine; thiocoraline;
thrombopoietin;
thrombopoietin mimetic; thymalfasin; thymopoietin receptor agonist;
thymotrinan; thyroid
stimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocene
bichloride; topsentin;
toremifene; translation inhibitors; tretinoin; triacetyluridine; triciribine;
trimetrexate;
triptorelin; tropisetron; turosteride; tyrosine kinase inhibitors;
tyrphostins; UBC inhibitors;
ubenimex; urogenital sinus-derived growth inhibitory factor; urokinase
receptor antagonists;
vapreotide; variolin B; velaresol; veramine; verdins; verteporfin;
vinorelbine; vinxaltine;
vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatin
stimalamer.
Specific second active agents include, but are not limited to, oblimersen
(GENASENSE , remicade, docetaxel, celecoxib, melphalan, dexamethasone
(DECADRONO), steroids, gemcitabine, cisplatinum, temozolomide, etoposide,
cyclophosphamide, temodar, carboplatin, procarbazine, gliadel, tamoxifen,
topotecan,
methotrexate, ARISA , taxol, taxotere, fluorouracil, leucovorin, irinotecan,
xeloda, CPT-11,
interferon alpha, pegylated interferon alpha (e.g., PEG INTRON-A),
capecitabine, cisplatin,
thiotepa, fludarabine, carboplatin, liposomal daunorubicin, cytarabine,
doxetaxol, pacilitaxel,
vinblastine, IL-2, GM-CSF, dacarbazine, vinorelbine, zoledronic acid,
palmitronate, biaxin,
busulphan, prednisone, bisphosphonate, arsenic trioxide, vincristine,
doxorubicin (DOXILO),
paclitaxel, ganciclovir, adriamycin, estramustine sodium phosphate (EMCYTO),
sulindac,
and etoposide.
In one embodiment, the compound or combination described herein may be used or
combined with one or more additional therapeutic agents. The one or more
therapeutic agents
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CA 3050553 2019-07-25
include, but are not limited to, an enzyme inhibitor, an inhibitor of Abl,
activated CDC kinase
(ACK), adenosine A2B receptor (A2B), apoptosis signal-regulating kinase (ASK),
Auroa
kinase, Bruton's tyrosine kinase (BTK), BET-bromodomain (BRD) such as BRD4, c-
Kit, c-
Met, CDK-activating kinase (CAK), calmodulin-dependent protein kinase (CaMK),
cyclin-
.. dependent kinase (CDK), casein kinase (CK), discoidin domain receptor
(DDR), epidermal
growth factor receptors (EGFR), focal adhesion kinase (FAK), Flt-3, FYN,
glycogen
synthase kinase (GSK), HCK, histone deacetylase (HDAC), IKK such as IKK3c,
isocitrate
dehydrogenase (IDH) such as IDH1, Janus kinase (JAK), KDR, lymphocyte-specific
protein
tyrosine kinase (LCK), lysyl oxidase protein, lysyl oxidase-like protein
(LOXL), LYN,
matrix metalloprotease (MMP), MEK, mitogen-activated protein kinase (MAPK),
NEK9,
NPM-ALK, p38 kinase, platelet-derived growth factor (PDGF), phosphorylase
kinase (PK),
polo-like kinase (PLK), phosphatidylinositol 3-kinase (PI3K), protein kinase
(PK) such as
protein kinase A, B, and/or C, PYK, spleen tyrosine kinase (SYK),
serine/threonine kinase
TPL2, serine/threonine kinase STK, signal transduction and transcription
(STAT), SRC,
serine/threonine-protein kinase (TBK) such as TBKI, TIE, tyrosine kinase (TK),
vascular
endothelial growth factor receptor (VEGFR), or any combination thereof.
Many experimental strategies for vaccination against cancers/tumors have been
devised (see
Rosenberg, S., 2000, Development of Cancer Vaccines, ASCO Educational Book
Spring: 60-
62; Logothetis, C, 2000, ASCO Educational Book Spring: 300-302; Khayat, D.
2000, ASCO
Educational Book Spring: 414-428; Foon, K. 2000, ASCO Educational Book Spring:
730-
738; see also Restifo, N. and Sznol, M., Cancer Vaccines, Ch. 61, pp. 3023-
3043 in DeVita,
V. et al. (eds.), 1997, Cancer: Principles and Practice of Oncology. Fifth
Edition). The study
of gene expression and large scale gene expression patterns in various tumors
has led to the
definition of so called tumor specific antigens (Rosenberg, S A (1999)
Immunity 10: 281-7).
In many cases, these tumor specific antigens are differentiation antigens
expressed in the
tumors and in the cell from which the tumor arose, for example melanocyte
antigens gp100,
MAGE antigens, and Trp-2.
In certain embodiments, a compound(s) of Formula [X] or a salt(s) thereof can
be
combined with one or more anti-cancer or anti-tumor vaccine, antigen,
immunogenic agent,
such as cancerous cells, purified tumor antigens (including recombinant
proteins, peptides,
and carbohydrate molecules), cells, and cells transfected with genes encoding
immune
stimulating cytokines (He et al (2004) J. Immunol. 173:4919-28). Non-limiting
examples of
tumor vaccines that can be used include peptides of melanoma antigens, such as
peptides of
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CA 3050553 2019-07-25
gp100, MAGE antigens, Trp-2, MARTI and/or tyrosinase, or tumor cells
transfected to
express the cytokine GM-CSF. Or for example, vaccines that secrete, or cause
the secretion
of, cytokines such as IL-2, SCF, CXC14 (platelet factor 4), G-CSF, and GM-CSF
can be used
in the methods, pharmaceutical compositions, and kits of the disclosure. See,
e.g., Emens, L.
A., et at., Curr. Opinion Mol. Ther. 3(1):77-84 (2001). The administered tumor
antigen may
also include the protein telomerase, which is required for the synthesis of
telomeres of
chromosomes and which is expressed in more than 85% of human cancers and in
only a
limited number of somatic tissues (Kim, Net al. (1994) Science 266: 2011-
2013).
Administered tumor antigen may also be "neo-antigens" expressed in cancer
cells because of
somatic mutations that alter protein sequence or create fusion proteins
between two unrelated
sequences (ie. bcr-abl in the Philadelphia chromosome), or idiotype from B
cell tumors.
Other tumor vaccines may include the proteins from viruses implicated in human
cancers
such a Human Papilloma Viruses (HPV), Hepatitis Viruses (HBV, HDV and HCV) and
Kaposi's Herpes Sarcoma Virus (KHSV). Administered antigen may be purified
heat shock
.. proteins (HSP) isolated from the tumor tissue itself. These heat shock
proteins contain
fragments of proteins from the tumor cells (Suot, R & Srivastava, P (1995)
Science 269:
1585-1588; Tamura, Y. et at. (1997) Science 278: 117-120). Dendritic cells
(DC) are potent
antigen presenting cells that can be used to prime antigen-specific responses
in a subject upon
administration. DCs can be produced ex vivo and loaded with various protein
and peptide
antigens as well as tumor cell extracts (Nestle, F. et at. (1998) Nature
Medicine 4: 328-332).
DCs may also be transduced by genetic means to express these tumor antigens as
well. DCs
have also been fused directly to tumor cells for the purposes of immunization
(Kugler, A. et
al. (2000) Nature Medicine 6:332-336).
In certain embodiments, a compound provided herein, e.g., the compound of
Formula [X], or
an enantiomer or a mixture of enantiomers thereof, or a pharmaceutically
acceptable salt,
solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is administered
in combination
with specific agents such as heparin, aspirin, coumadin, or G-CSF to avoid
adverse effects
that are associated with anti-cancer drugs such as but not limited to
neutropenia or
thrombocytopenia.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula [X], or
an enantiomer or a mixture of enantiomers thereof, or a pharmaceutically
acceptable salt,
solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is administered
with melphalan
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CA 3050553 2019-07-25
and dexamethasone to patients with amyloidosis. In certain embodiments, a
compound
provided herein, e.g., the compound of Formula [X], or an enantiomer or a
mixture of
enantiomers thereof, or a pharmaceutically acceptable salt, solvate, hydrate,
co-crystal,
clathrate, or polymorph thereof, and steroids can be administered to patients
with
amyloidosis.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
= [X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered with
gemcitabine and cisplatinum to patients with locally advanced or metastatic
transitional cell
bladder cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with a second active ingredient as follows: temozolomide to
pediatric patients
with relapsed or progressive brain tumors or recurrent neuroblastoma;
celecoxib, etoposide
and cyclophosphamide for relapsed or progressive CNS cancer; temodar to
patients with
recurrent or progressive meningioma, malignant meningioma, hemangiopericytoma,
multiple
brain metastases, relapsed brain tumors, or newly diagnosed glioblastoma
multiforms;
irinotecan to patients with recurrent glioblastoma; carboplatin to pediatric
patients with brain
stem glioma; procarbazine to pediatric patients with progressive malignant
gliomas;
cyclophosphamide to patients with poor prognosis malignant brain tumors, newly
diagnosed
or recurrent glioblastoma multiforms; Gliadel for high grade recurrent
malignant gliomas;
temozolomide and tamoxifen for anaplastic astrocytoma; or topotecan for
gliomas,
glioblastoma, anaplastic astrocytoma or anaplastic oligodendroglioma.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered with
methotrexate, cyclophosphamide, taxane, abraxane, lapatinib, herceptin,
aromatase inhibitors,
selective estrogen modulators, estrogen receptor antagonists, and/or PLX3397
(Plexxikon) to
patients with metastatic breast cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered with
temozolomide to patients with neuroendocrine tumors.
530
CA 3050553 2019-07-25
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered with
gemcitabine to patients with recurrent or metastatic head or neck cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered with
gemcitabine to patients with pancreatic cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered to patients
with colon cancer in combination with ARISAS, avastatin, taxol, and/or
taxotere.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered with
capecitabine and/or PLX4032 (Plexxikon) to patients with refractory colorectal
cancer or
patients who fail first line therapy or have poor performance in colon or
rectal
adenocarcinoma.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with fluorouracil, leucovorin, and irinotecan to patients with
Dukes C & D
colorectal cancer or to patients who have been previously treated for
metastatic colorectal
cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered to patients
with refractory colorectal cancer in combination with capecitabine, xeloda,
and/or CPT-11.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered with
capecitabine and irinotecan to patients with refractory colorectal cancer or
to patients with
unresectable or metastatic colorectal carcinoma.
531
CA 3050553 2019-07-25
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered alone or in
combination with interferon alpha or capecitabine to patients with
unresectable or metastatic
hepatocellular carcinoma; or with cisplatin and thiotepa to patients with
primary or metastatic
liver cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with pegylated interferon alpha to patients with Kaposi's sarcoma.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with fludarabine, carboplatin, and/or topotecan to patients with
refractory or
relapsed or high-risk acuted myelogenous leukemia.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with liposomal daunorubicin, topotecan and/or cytarabine to
patients with
unfavorable karotype acute myeloblastic leukemia.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with gemcitabine, abraxane, erlotinib, geftinib, and/or irinotecan
to patients with
non-small cell lung cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with carboplatin and irinotecan to patients with non-small cell
lung cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered with
doxetaxol to patients with non-small cell lung cancer who have been previously
treated with
carboNP 16 and radiotherapy.
532
CA 3050553 2019-07-25
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with carboplatin and/or taxotere, or in combination with
carboplatin, pacilitaxel
and/or thoracic radiotherapy to patients with non-small cell lung cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with taxotere to patients with stage IIIB or IV non-small cell
lung cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with oblimersen (Genasensee) to patients with small cell lung
cancer.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with ABT-737 (Abbott Laboratories) and/or obatoclax (GX15-070) to
patients
with lymphoma and other blood cancers.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered alone or in
combination with a second active ingredient such as vinblastine or fludarabine
to patients
with various types of lymphoma, including, but not limited to, Hodgkin's
lymphoma, non-
Hodgkin's lymphoma, cutaneous T-Cell lymphoma, cutaneous B-Cell lymphoma,
diffuse
large B-Cell lymphoma or relapsed or refractory low grade follicular lymphoma.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered in
combination with taxotere, IL-2, IFN, GM-CSF, PLX4032 (Plexxikon) and/or
dacarbazine to
patients with various types or stages of melanoma.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered alone or in
combination with vinorelbine to patients with malignant mesothelioma, or stage
IIIB non-
533
CA 3050553 2019-07-25
small cell lung cancer with pleural implants or malignant pleural effusion
mesothelioma
syndrome.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered to patients
with various types or stages of multiple myeloma in combination with
dexamethasone,
zoledronic acid, palmitronate, GM-CSF, biaxin, vinblastine, melphalan,
busulphan,
cyclophosphamide, IFN, palmidronate, prednisone, bisphosphonate, celecoxib,
arsenic
trioxide, PEG INTRON-A, vincristine, or a combination thereof.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered to patients
with relapsed or refractory multiple myeloma in combination with doxorubicin
(Doxile),
vincristine and/or dexamethasone (Decadrone).
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered to patients
with various types or stages of ovarian cancer such as peritoneal carcinoma,
papillary serous
carcinoma, refractory ovarian cancer or recurrent ovarian cancer, in
combination with taxol,
carboplatin, doxorubicin, gemcitabine, cisplatin, xeloda, paclitaxel,
dexamethasone, or a
combination thereof.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered to patients
with various types or stages of prostate cancer, in combination with xeloda, 5
FU/LV,
gemcitabine, irinotecan plus gemcitabine, cyclophosphamide, vincristine,
dexamethasone,
GM-CSF, celecoxib, taxotere, ganciclovir, paclitaxel, adriamycin, docetaxel,
estramustine,
Emcyt, denderon or a combination thereof.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered to patients
with various types or stages of renal cell cancer, in combination with
capecitabine, IFN,
tamoxifen, IL-2, GM-CSF, Celebrex , or a combination thereof.
534
CA 3050553 2019-07-25
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered to patients
with various types or stages of gynecologic, uterus or soft tissue sarcoma
cancer in
combination with IFN, a COX-2 inhibitor such as Celebrex , and/or sulindac.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered to patients
with various types or stages of solid tumors in combination with celebrex,
etoposide,
cyclophosphamide, docetaxel, apecitabine, IFN, tamoxifen, IL-2, GM-CSF, or a
combination
thereof.
In certain embodiments, a compound provided herein, e.g., the compound of
Formula
[X], or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically acceptable
salt, solvate, hydrate, co-crystal, clathrate, or polymorph thereof, is
administered to patients
.. with scleroderma or cutaneous vasculitis in combination with celebrex,
etoposide,
cyclophosphamide, docetaxel, apecitabine, IFN, tamoxifen, IL-2, GM-CSF, or a
combination
thereof.
The above "other" therapeutic agents, when employed in combination with a
compound(s) of
the present disclosure, may be used, for example, in those amounts indicated
in the
Physicians' Desk Reference (PDR) or as otherwise determined by one of ordinary
skill in the
art. In the methods of the present disclosure, such other therapeutic agent(s)
may be
administered prior to, simultaneously with, or following the administration of
the inventive
compounds.
Use with transplantation therapy
The abovementioned therapies can be supplemented or combined with stem cell
transplantation or treatment, including peripheral blood stem cell
transplantation, autologous
hematopoietic stem cell transplantation, autologous bone marrow
transplantation, infusion of
stem cells, bone marrow ablation with stem cell support, in vitro-treated
peripheral blood
stem cell transplantation, umbilical cord blood transplantation, and
nonmyeloablative
allogeneic hematopoietic stem cell transplantation.
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CA 3050553 2019-07-25
The compound(s) of Formula [X] provided herein can be used to reduce the risk
of Graft
Versus Host Disease (GVHD). Therefore, encompassed herein is a method of
treating,
preventing and/or managing cancer, which comprises administering one or more
of these
compounds, or an enantiomer or a mixture of enantiomers thereof, or a
pharmaceutically
acceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorph
thereof, in conjunction
with transplantation therapy.
As those of ordinary skill in the art are aware, the treatment of cancer is
often based on
the stages and mechanism of the disease. For example, as inevitable leukemic
transformation
develops in certain stages of cancer, transplantation of peripheral blood stem
cells,
hematopoietic stem cell preparation or bone marrow may be necessary. The
combined use of
a compound provided herein and transplantation therapy provides a unique and
unexpected
synergism. In particular, the compound exhibits immunomodulatory activity that
may provide
additive or synergistic effects when given concurrently with transplantation
therapy in
patients with cancer.
The compound(s) of Formula [X] can work in combination with transplantation
therapy
reducing complications associated with the invasive procedure of
transplantation and risk of
GVHD. Encompassed herein is a method of treating, preventing and/or managing
cancer
which comprises administering to a subject at least one compound of Formula
[X], or an
enantiomer or a mixture of enantiomers thereof, or a pharmaceutically
acceptable salt,
solvate, hydrate, co-crystal, clathrate, or polymorph thereof, before, during,
or after the
transplantation of umbilical cord blood, placental blood, peripheral blood
stem cell,
hematopoietic stem cell preparation, or bone marrow. Some examples of stem
cells suitable
for use in the methods provided herein are disclosed in U.S. Pat. No.
7,498,171, the
disclosure of which is incorporated herein by reference in its entirety.
In one embodiment, at least one compound of Formula [X] is administered to
patients
with multiple myeloma before, during, or after the transplantation of
autologous peripheral
blood progenitor cell.
In another embodiment, at least one compound of Formula [X] is administered to
patients
with relapsing multiple myeloma after the stem cell transplantation.
In yet another embodiment, at least one compound of Formula [X] and prednisone
are
administered as maintenance therapy to patients with multiple myeloma
following the
transplantation of autologous stem cell.
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CA 3050553 2019-07-25
In yet another embodiment, at least one compound of Formula [X] and
dexamethasone
are administered as salvage therapy for low risk post transplantation to
patients with multiple
myeloma.
In yet another embodiment, at least one compound of Formula [X] and
dexamethasone
are administered as maintenance therapy to patients with multiple myeloma
following the
transplantation of autologous bone marrow.
In yet another embodiment, at least one compound of Formula [X] is
administered
following the administration of high dose of melphalan and the transplantation
of autologous
stem cell to patients with chemotherapy responsive multiple myeloma.
In yet another embodiment, at least one compound of Formula [X] and PEG INTRO-
A
are administered as maintenance therapy to patients with multiple myeloma
following the
transplantation of autologous CD34-selected peripheral stem cell.
In yet another embodiment, at least one compound of Formula [X] is
administered with
post transplant consolidation chemotherapy to patients with newly diagnosed
multiple
myeloma to evaluate anti-angiogenesis.
In still another embodiment, at least one compound of Formula [X] and
dexamethasone
are administered as maintenance therapy after DCEP consolidation, following
the treatment
with high dose of melphalan and the transplantation of peripheral blood stem
cell to 65 years
of age or older patients with multiple myeloma.
In one embodiment at least one compound of Formula [X] is administered to
patients with
NHL (e.g., DLBCL) before, during, or after the transplantation of autologous
peripheral
blood progenitor cell.
In another embodiment, at least one compound of Formula [X] is administered to
patients
with NHL (e.g., DLBCL) after a stem cell transplantation.
Scope
The examples of this invention are provided to better illustrate the claimed
invention and are
not to be interpreted in any way as limiting the scope of the invention. These
specific
compounds, compositions, materials, methods and kits are not intended to limit
the invention,
but merely to illustrate specific embodiments falling within the scope of the
invention. One
skilled in the art may develop equivalent compounds, compositions, materials,
and methods
without the exercise of inventive capacity and without departing from the
scope of the
invention. It is therefore to be understood that within the scope of the
appended claims, the
invention may be practiced otherwise than as specifically described herein. It
will be
537
CA 3050553 2019-07-25
understood that many variations can be made in the procedures herein described
while still
remaining within the bounds of the invention. It is the intention of the
inventor that such
variations are included within the scope of the invention. The present
invention may be
embodied in other specific forms without departing from the spirit or
essential attributes
thereof. This invention encompasses all combinations of preferred aspects of
the invention
noted herein. It is understood that any and all embodiments of the present
invention may be
taken in conjunction with any other embodiment or embodiments to describe
additional
embodiments. It is also to be understood that each individual element of the
embodiments is
its own independent embodiment. Furthermore, any element of an embodiment is
meant to be
combined with any and all other elements from any embodiment to describe an
additional
embodiment. Feature(s) described in connection with one embodiment of the
invention may
be used in conjunction with another embodiment(s), even if not explicitly
stated.
Abbreviations
All abbreviations used in this disclosure are standard/used in the art,
familiar/discernable to a
person of the art, especially in their context, to illustrate:
Ph=phenyl
Bn=benzyl
Me=methyl
Et=ethyl
Me0H=methanol
Et0H=ethanol
Pr=propyl
Bu=butyl
PE=petroleum ether
COOEt=ethoxycarbonyl
CO2Et=ethoxycarbonyl
Et3SiH=triethylsilane
LiA1H4=lithium aluminium hydride
ACN=acetonitrile=CH3CN
AcOH=acetic acid
MeI=CH3I
Boc=tert-butyloxycarbonyl protecting group
538
CA 3050553 2019-07-25
CDI=1,1'-carbonyldiimidazole
DCE=1,2-Dichloroethane
DCM=dichloromethane
DBU=1,8-diazabicyclo[5,4,0]undec-7-ene
DMAP=4-Dimethylaminopyridine
DMF=N,N-dimethylformamide
DPPA=Diphenylphosphoryl azide
EDCI=N-(3-dimethylaminopropy1)-N'-ethylcarbodiimide hydrochloride
Et0Ac=ethyl acetate
HEPES= 4-(2-hydroxyethyl)- 1 -piperazineethanesulfonic acid
IBX=2-iodoxybenzoic acid
IPA=isopropanol
MEI=methyl iodide
Tf20=triflic anhydride
NaBH(OAc)3=sodium triacetoxyborohydride
NCS= N=C=S
Rf=retention value
t-BuOK=potassium tert-butoxide
TEA=triethylamine=Et3N
THF=tetrahydrofuran
TFA=trifluoroacetic acid
T3P=Propanephosphonic acid anhydride
HATU=Hexafluorophosphate Azabenzotriazole Tetramethyl Uronium
TLC=thin-layer chromatography
SFC=supercritical fluid chromatography or chiral supercritical fluid
chromatography
IPAm=isopropylamine
SM=starting material
min=minute(s)
h or hr=hour(s)
aq. or aq=aqueous
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Green D,
Ferrara FN, inventors; Bristol-Myers Squibb Co., assignee. (1-phenyl-2-
heteoaryl) ethyl-
539
CA 3050553 2019-07-25
guanidine compounds as inhibitors of mitochondrial FIFO ATP hydrolase. United
States
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[P2] Ding C, Hamann L, Stein P, Pudzianowski A, inventors; Ding Charles Z.,
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[P5] Glick G, inventor; University of Michigan, assignee. Methods and
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[P6] Ding CZ, Atwal KS, inventors; Bristol-Myers Squibb Company, assignee.
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[P7] Regnier G, Canevari R, Laubie M, inventors; En Nom Collectif Science
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NN'substituted polymethylene diamines. United States patent US 4,514,399. 1985
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[377] Mitsui A, Hamuro J, Nakamura H, Kondo N, Hirabayashi Y, Ishizaki-Koizumi
S,
Hirakawa T, Inoue T, Yodoi J. Overexpression of human thioredoxin in
transgenic mice
controls oxidative stress and life span. Antioxidants and Redox Signaling.
2002 Aug
1 ;4(4):693-6.
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CA 3050553 2019-07-25
DESCRIPTION claims (Method of use format)
[1] A compound, or a composition containing at least one compound, of the
following
formula:
R2NN/R3
=N
L
HI
(R4)q R1
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
L is alkyl, or deuterium, or substituted alkyl, or deuterated alkyl, or
aminoalkyl, or
thioalkyl, or alkoxy, or halogen, or haloalkyl, or haloalkoxy, or
hydroxyalkyl, or any atom or
isotope permitted by valence except hydrogen at natural abundance;
RI is hydrogen, cyano, ¨SO2R8, ¨C(3)R9, heteroaryl or thiazolyl;
R2 is (i) independently hydrogen, alkyl, benzyl, or substituted alkyl, or (ii)
taken
together with R3 forms a heterocyclo;
R3 is (i) independently alkyl, substituted alkyl, alkylthio, aminoalkyl,
carbamyl, BB-
aryl, BB-heterocyclo, BB-heteroaryl, or BB-cycloalkyl, or (ii) phenyl
optionally substituted
with Ci..4alkyl, halogen, trifluoromethyl, OCF3, cyano, nitro, amino, hydroxy,
or methoxy, or
(iii) independently selected from Ci_aalkyl, alkylthio, aminoalkyl, -BB-aryl, -
BB-heterocyclo,
BB-cycloalkyl, and -BB-hetaroaryl, optionally having one to three substituents
selected from
R3a; and/or having fused thereto a five or six membered carbocyclic ring, or
(iv) taken
together with R2 forms a heterocyclo optionally substituted with alkyl or
substituted alkyl;
BB is a bond, Chaalkylene, C2_4alkenylene, substituted Ci_aalkylene,
substituted C2-
4a1keny1ene, substituted Ci_4alkylene-C(=0)NH¨, ¨Ci_4alkylene-
C(=0)NH¨, ¨C(I)NRI ¨C
malkylene-C()NRI9¨, or substituted C 4alkylene-
C(=0)NRI9¨, ¨(CHR14)m¨(CRI5R16),¨ or ¨(CHR14)p¨C(=0)NH¨;
R3a at each occurrence is selected independently from alkyl, substituted
alkyl,
halogen, haloalkoxy, cyano, nitro, keto, trifluoromethyl, ¨NRI7R18, ¨SR17,
¨0R17, ¨
-- SO2R17a, ¨S02NRI7R18, ¨Nit17g=a)R18, --0O2R17, --C(D)R17, cyoloalkyl, aryl,
heterocyolo, and heteroaryl, wherein when R3a is cycloalkyl, aryl, heterocyclo
or heteroaryl,
said cycloalkyl, aryl, heterocyolo and heteroaryl in turn is optionally
substituted with alkyl or
substituted alkyl;
576
CA 3050553 2019-07-25
Z is a heteroaryl, for example an optionally-substituted bicyclic heteroaryl;
or
Z is triazolyl optionally substituted with one to two R7 substituents or
imidazolyl
optionally substituted with one to two R7 substituents and/or having fused
thereto a benzene
ring in turn optionally substituted with one to two R7 substituents; and
R7 is alkyl, carbamyl, or substituted alkyl;
R4 at each occurrence is selected independently of each other R4 from the
group
consisting of halogen, trifluoromethyl, OCF3, alkyl, substituted alkyl,
haloalkyl, nitro, cyano,
haloalkoxy, OR25, SR25, NR25R26, NR25S02R27, S02R27, S02NR25R26, CO2R26,
C(=0)R26,
C(=0)NR25R26, OC(=C))R25, -0C(=0)NR25R26, NR25C(:))R26, NR25CO2R26, aryl,
heteroaryl, heterocyclo and cycloalkyl;
R8 is Ci_aalkyl or phenyl optionally substituted with alkyl, halogen,
haloalkoxy, cyano,
nitro, or trifluoromethyl;
R9 is -NRIORII, alkyl, substituted alkyl, alkoxy, alkylthio, cycloalkyl, aryl,
heteroaryl, heterocyclo, or -0O2R12, alkyl or phenyl optionally substituted
with one to four
of halogen, cyano, trifluoromethyl, nitro, hydroxy, Ci_aalkoxy, haloalkoxy,
C1_6alkyl,
CO2alkyl, SO2alkyl, SO2NH2, amino, NH(C1-4alkyl), N(Ci_4alky1)2, NHC(=0)alkY,
C(=0)alkyl, and/or Ci_aalkyl optionally substituted with one to three of
trifluoromethyl,
hydroxy, cyano, phenyl, pyridinyl; and/or a five or six membered heteroaryl or
heterocylo in
turn optionally substituted with keto or having a benzene ring fused thereto
or
a) Ci_aalkyl optionally substituted with one to two of:
i) SR13, 0R13, NR13aRi3b, halogen, trifluoromethyl, CO2R13a, and
C(=0)NR13aRi3b;
ii) cycloalkyl optionally substituted with one to two of C(=0)H, Ci_aacyl,
alkenyl, carbamyl,
and/or phenyl in turn optionally substituted with halogen;
iii) phenyl or napthyl optionally substituted with one to two of halogen,
nitro, amino, alkyl,
hydroxy, Ci_aalkoxy, or having fused thereto a five or six membered
heterocyclo;
iv) pyridinyl, thiophenyl, fiiranyl, tetrahydrofuranyl, or azepinyl,
optionally substituted with
alkyl or having fused thereto a five to six membered carbocyclic ring
optionally substituted
with keto or Ci_4a1k0xy;
b) 3 to 6 membered cycloalkyl optionally having up to four substituetits
selected from alkyl,
-- halogen, cyano, alkenyl, acyl, alkylthio, carbamyl, phenyl in turn
optionally substituted with
halogen; or having an aryl fused thereto;
c) pheyl optionally substituted with one to four of halogen, cyano,
trifluoromethyl, nitro,
hydroxy, Ci_aalkoxy, haloalkoxy, Ci_aalkyl, CO2alkyl, SO2alkyl, SO2NH2, amino
NH(Ci_
4a1ky1), N(Ci_4alky1)2, NHC(=3)alkyl, C(=;30)alkyl, and/or Chaalkyl optionally
substituted
577
CA 3050553 2019-07-25
with one to three of trifluoromethyl, hydroxy, cyano, phenyl, pyridinyl;
and/or a five or six
membejed heteroaryl or heterocyle in turn optionally substituted with keto or
having a
benzene ring fused thereto;
d) pyridinyl, thiazolyl, furanyl, thiophenyl, and pyrrolyl optionally
substituted with one to
two of halogen, alkyl, and phenyl in turn optionally substituted with halogen
or
trifluoromethyl;
Rio and Ri I are (i) independently selected from hydrogen, alkyl, substituted
alkyl,
alkoxy, heterocyclo, cycloalkyl, aryl, heteroaryl or Ci_4alkyl optionally
substituted with one
to two of ¨0O2alkyl, ¨C(C)NH(ary1), NH(ary1), cycloalkyl, phenyloxy, phenyl in
turn
optionally substituted with Ci_aalkyl, hydroxy, Ci_aalkoxy, halogen, amino,
nitro,
tetrahydrofuranyl, and/or five or six membered heterocyclo, or having a five
or six membered
heterocyclo fused thereto; pyrrolidinyl optionally substituted with keto;
napthyl, anthracenyl,
pyridinyl, thiophenyl, furanyl, imidazolyl, benzimidazolyl, or indolyl in turn
optionally
substituted with Ci_4alkyl or Ci_aalkoxy; or (ii) taken together form a
heteroaryl or
heterocyclo selected from pyrrolidinyl, piperazinyl, piperidinyl, morpholinyl,
tetrahydropyridinyl, and imidazoilidinyl, wherein said heterocyclo formed by
Rio and RH is
optionally substituted with one to two of keto, CO2H, Ciaalkoxy, CO2alky1,
Ci_acarbamyl,
benzyl; phenyl in turn optionally substituted with alkyl, halogen, or CI-4
alkoxy;
tetrahydropyridinyl in turn optionally substituted with keto and/or phenyl;
alkyl optionally
substituted with amino or NHR21wherein R21 is alkyl or phenyl optionally
substituted with
alkyl; and/or has a benzene ring fused thereto in turn optionally substituted
with one to two of
alkyl, Ci_aalkoxy, CO2alkyl, and/or Ci_acarbamyl;
R12 and R19 are hydrogen or alkyl;
Ri3 is hydrogen or alkyl;
Ri3a and Ri3b are selected from hydrogen, alkyl, and aryl;
RI4, RI5 and RI6 at each occurrence are independently selected from hydrogen,
alkyl,
hydroxy, hydroxyCi_aalkyl, Cmalkoxy, and phenyl, and/or one of R15 and one of
Ri6 join
together to form a 3 to 6 membered cycloalkyl;
R17 and R18 are independently selected from hydrogen, alkyl, substituted
alkyl, aryl,
phenyl, or benzyl wherein the phenyl or benzyl is optionally substituted with
alkyl, hydroxy,
or hydroxyalkyl;
RIM is alkyl or substituted alkyl;
R25 and R26 are independently selected from hydrogen, alkyl, or substituted
alkyl, or
taken together form a heterocyclo or heteroaryl ring;
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CA 3050553 2019-07-25
R27 is alkyl or substituted alkyl;
q is 0, 1, 2, or 3;
m and n are 0, 1 or 2; and
p is 0, 1, 2, or 3.
[2] A compound according to Claim 1, of the formula,
R2 R3
0,s\L
R1
(R4),14*
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
there is an enantiomeric excess (cc) of the particular stereoisomer shown in
the
formula.
[3] A compound according to Claim 1, of the formula,
R2 R3
NN./
N/L
N
D
R1
(R4)q
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
D is deuterium (enrichment, for (non-limiting) example, exceeding 40%
deuterium
incorporation at position shown, optionally deuterium enrichment at other
locations also),
141 A compound according to Claim 1, of the formula,
579
CA 3050553 2019-07-25
R3
Z R2N r
N
,oxµD
Slik S
H
I
(R4)q R1
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
D is deuterium (enrichment, for example, exceeding 40% deuterium incorporation
at
position shown);
the enantiomeric excess (ee) of the S stereoisomer exceeds 70%;
[5] A compound according to Claim 1, of the formula,
Z R2N. ..,R3
N
ZIN\
jjkN " N
CH3 H I
R1
(R4)q
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof.
[6] A compound according to Claim 1, of the formula,
Z R2 R3
N
NV
ACH>IN
ss"
N X N
I
R1
(R4)q
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
the enantiomeric excess (ee) of the S stereoisomer exceeds 70%;
[7] A compound according to Claim 1, of the formula,
580
CA 3050553 2019-07-25
R2 R3
Z X N ./.
CI*
(R4)q RD'f// N ga
N N
I
H
R1
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
the enantiomeric excess (ee) of the R stereoisomer exceeds 70%;
-- 181 A compound according to Claim 1, wherein,
R7c
R7a .............c,N,R7b R7a ...,.......\,k,N
or \
Y ¨N N
c
Y is CH, N or CIZ7c;
RI is cyano, ¨S02R8, ¨C(=D)R9, or heteroaryl;
R2 is (i) independently hydrogen, alkyl, or substituted alkyl, or (ii) taken
together with
R3 forms a heterocyclo;
R3 is (i) independently selected from
(a) alkyl optionally substituted with one to two of hydroxy and alkoxy;
(b) alkylthio or aminoalkyl optionally substituted with hydroxy or alkoxy;
(c) -At-aryl, wherein the aryl is optionally substituted with up to four
substituents selected
from alkyl, substituted alkyl, halogen, haloalkoxy, cyano, nitro, ¨NRI7R18,
¨SR17, ¨0R17,
¨S021Zi7a, ¨S02NRI7R18, ¨NRi7C(=a)R18, ¨0O2R17, ¨C(=0)R17, cycloalkyl, aryl,
heterocyclo, and heteroaryl, and/or has fused thereto a five or six membered
cycloalkyl ring;
(d) -A2-heteroaryl wherein the heteroaryl is a five or six membered monocyclic
ring having 1
to 3 heteroatoms selected from N, 0, and S, or an eight or nine membered
bicyclic ringed
system having at least one aromatic ring and 1 to 4 heteroatoms selected from
N, 0, and S in
at least one of the rings, said heteroaryl being optionally substituted with
halogen. alkyl,
alkoxycarbonyl, sulfonamide, nitro, cyano, trifluoromethyl, alkylthio, alkoxy,
keto, ¨
581
CA 3050553 2019-07-25
C(=0)H, acyl, benzyloxy, hydroxy, hydroxyalkyl, or phenyl optionally
substituted with alkyl
or substituted alkyl;
(e) -A2-heterocyclo wherein the heterocyclo is optionally substituted with one
to two groups
selected from alkyl, keto, hydroxy, hydroxyalkyl, ¨C(=0)H, acyl, CO2H,
alkoxycarbonyl,
phenyl, and/or benzyl, and/or has a bridged carbon¨carbon chain or fused
benzene ring
joined thereto;
(0 -A2-cycloalkyl wherein the cycloalkyl is optionally substituted with one to
two groups
selected from alkyl, keto, ¨C(=0)H, acyl, CO2H, alkoxycarbonyl, and/or benzyl,
and/or has
a bridged carbon¨carbon chain or fused benzene ring joined thereto; or
(ii) taken together with R2 forms a heterocyclo;
R4 at each occurrence is selected independently of each other R4 from the
group
consisting of halogen, alkyl, haloalkyl, cyano, and haloalkoxy;
R7a, R7b and R7c are independently selected from hydrogen, alkyl, carbamyl, or
carbamylalkyl, or R7a and R7c join to form an aryl or heteoraryl;
R8 is alkyl, arylalkyl, or aryl;
R9 is ¨NRIORI I, alkyl, substituted alkyl, alkoxy, alkylthio, cycloalkyl,
aryl,
heteroaryl, heterocyclo, CO2R12, or phenyl optionally substituted with one to
four of halogen,
cyano, trifluoromethyl, nitro, hydroxy, Ci_aalkoxy, haloalkoxy, Ci_6alkyl,
CO2alkyl, SO2alkyl,
SO2NH2, amino, NH(C1.4alkyl), N(Ci_4alky1)2, NHC(=0)alkyl, C(=0)alkyl, and/or
Ci_aalkyl
optionally substituted with one to three of trifluoromethyl, hydroxy, cyano,
phenyl, pyridinyl;
and/or a five or six membered heteroaryl or heterocyclo in turn optionally
substituted with
keto or having a benzene ring fused thereto;
Rio is independently hydrogen, alkyl, or alkoxy; and
Ri is independently hydrogen, alkyl, substituted alkyl, alkoxy heterocyclo
cycloalkyl,
aryl, or heteroaryl; or
Rio and Ru1 taken together form a heterocyclo or heteroaryl optionally
substituted with
alkyl, keto, CO2H, alkoxycarbonyl, hydroxy, alkoxy, alkyl, carbamyl, aryl, or
substituted
alkyl, wherein when the RIO and Ri group comprises a phenyl ring, said phenyl
ring is
optionally substituted with one to two of alkyl, halogen, and alkoxy;
Ri2 is hydrogen or alkyl;
Au is ¨(CHRia)m¨V¨(CRI5R16)n¨ or ¨(CHR14)p¨(C=D)NH¨;
A2 IS ¨(CHR14)m¨V¨(CRI5R16)n;
V is a bond, S, or ¨NR22¨;
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CA 3050553 2019-07-25
Ri4, RI5 and R16 at each occurrence are independently selected from hydrogen,
alkyl,
hydroxy, hydroxyCi-aalkyl, Ci_aalkoxy, and phenyl, and/or one of R15 and one
of Ri6join
together to form a three to six membered cycloalkyl;
R17 and Riaare independently selected from hydrogen, alkyl, phenyl, and
benzyl,
wherein the phenyl and benzyl is optionally substituted with alkyl, hydroxy,
or hydroxyalkyl;
R17a is alkyl or substituted alkyl;
R22 is hydrogen or alkyl;
m and n are 0, 1, 2, or 3;
p is 0, 1, 2, or 3; and
q is 0, 1, 2, or 3.
191 A compound according to Claim 8 having the formula,
(R24)y
41111
Z NNZ R2 (CHR23)x
L
NrkN
H
I
(R4) R1
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
R7a, R7b and R7c are independently selected from hydrogen, alkyl, carbamyl or
carbamylCi_aalkyl, or R7a and R7, join to form a fused phenyl ring;
R23 is selected from hydrogen, alkyl, hydroxyalkyl, or phenyl;
R24 is selected from alkyl, halogen, trifluoromethyl, cyano, halogen, hydroxy,
OCF3,
methoxy, phenyloxy, benzyloxy, cyano, acyl, or two R24 groups join to form a
fused
cycloalkyl or benzene ring; and
x is 0, 1, or 2; and
y is 0, 1, 2, or 3.
1101 A compound according to Claim 1, having the structure:
583
CA 3050553 2019-07-25
N
1110
N/
0
CI
=
CI 1110
HZNNH
4111
CI
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
deuterium (D) enrichment at the chiral centre, and optionally at other
locations,
exceeds 40% deuterium incorporation
the enantiomeric excess (ee) of the S stereoisomer exceeds 70%.
1111 A compound according to Claim 1, having the structure:
584
CA 3050553 2019-07-25
......==" N
N'('7%'
t¨N
N 0
CI CH3A
CI * N
H NH
0
Ci
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof.
1121 A compound according to Claim 1, having the structure:
<21
1.1
NiZZL--1
N
N 0
CI AC H3 I
el s ....................
N NH
H
CI
01
CI
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
the enantiomeric excess (ee) of the S stereoisomer exceeds 70%.
585
CA 3050553 2019-07-25
1131 A compound according to Claim 1, having the structure:
N
11/\
NO
CI CH3/1
Rs"//N NH
CI
411
CI
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
the enantiomeric excess (ee) of the R stereoisomer exceeds 70%.
[14] A pharmaceutical composition comprising at least one compound as defined
in Claim 1
and a pharmaceutically-acceptable carrier or excipient or diluent.
[15] At least one compound, or a composition containing at least one compound,
according to
Claim 1 for use in a method of treatment of the human or animal body by
therapy.
[16] A method of treating, ameliorating, preventing, reversing or combating a
disease or
disorder, or unwanted/undesirable physiological process or its consequences or
an
unwanted/undesirable aesthetic, in a subject, selected from
(i) cancer that metabolizes much of its glucose and/or glutamine to lactate,
for
example a cancer exhibiting the Warburg effect and/or a cancer that can be
discriminated
from surrounding tissue by PET imaging (e.g. 18F-FDG PET);
(ii) cachexia, cancer driven cachexia, cancer fatigue, weight loss, weight
loss for
known or unknown reason, chronic wasting disease, atrophy, brown atrophy,
frailty, frailty
586
CA 3050553 2019-07-25
syndrome, aging frailty, geriatric syndrome, age-related cachexia and/or
sarcopenia,
weakness, wasting, anorexia nervosa, bulimia nervosa, eating disorder,
amenorrhea,
underweight, low body mass index (BMI, e.g. <18.5), low body fat percentage,
body
composition change, wasting syndrome, HIV wasting syndrome, malnutrition,
clinical
.. malnutrition, starvation, kwashiorkor syndrome, marasmus syndrome,
malabsorption,
malabsorption due to parasitic/bacterial infection (e.g. helminthiasis,
Whipple's disease, small
intestine bacterial overgrowth (SIB0), giardiasis etc.), anemia, refeeding
syndrome, appetite
loss, catabolysis, muscle atrophy, asthenia, muscle weakness (myasthenia),
sarcopenia,
osteoporosis, cachexia associated with HIV, AIDS, multiple sclerosis,
rheumatoid arthritis,
familial amyloid polyneuropathy, chronic kidney disease, cystic fibrosis,
multiple sclerosis,
motor neuron disease, Parkinson's disease, dementia, Addison's disease,
mercury poisoning
(acrodynia), chronic pancreatitis, untreated/severe type 1 diabetes mellitus,
hormonal
deficiency, tuberculosis, gastroenteritis, diarrhea, dysentery, any digestive
disease or
disorder, any gastrointestinal disease or disorder including functional
gastrointestinal
disorders, coeliac disease, tropical sprue, irritable bowel syndrome,
inflammatory bowel
disease, Crohn's disease, ulcerative colitis, short bowl syndrome, congestive
heart failure,
constrictive pericarditis, bradycardia, chronic obstructive pulmonary disease
(COPD),
altitude sickness, hyperthyroidism (subclinical hyperthyroidism, Graves'
disease,
multinodular goiter, toxic adenoma, inflammation of the thyroid {thyroiditis},
pituitary
adenoma), fatigue, chronic fatigue syndrome or any disease or disorder or
pathology in which
a body tissue(s) is undersupplied or underutilises (vs. its need) an
energetic/chemical
substrate(s), including 02;
(iii) cancer associated fever, which is especially associated with, but not
limited to,
non Hodgkin lymphoma (NHL), Hodgkin lymphoma, acute leukaemia, kidney cancer
(renal
cell cancer), liver cancer (hepatocellular carcinoma), bone cancer, adrenal
gland tumours
such as phaeochromocytomas, tumours in the hypothalamus, solid tumours;
(iv) disease or disorder or physiological process or condition that causes a
higher than
normal body temperature such as (without limitation) high environmental
temperature,
ingesting an uncoupler (e.g. 2,4-dinitrophenol), infection, sepsis,
neutropenic sepsis, stroke,
.. fever, pyrexia, hyperpyrexia, hyperthermia, malignant hyperthermia,
neuroleptic malignant
syndrome, serotonin syndrome, toxic serotonin syndrome, thyroid storm,
heatstroke, surgery
related, menopause ("hot flushes"), infection (non-limiting e.g. roseola,
measles, enteroviral
infections, parasitic, viral, fungal, Chlamydial, Rickettsial, bacterial,
mycobacterial, systemic
bacterial, intravascular, 1-11V associated, nosocomial), pyrogenic infection,
thermoregulatory
587
CA 3050553 2019-07-25
disorder(s), connective tissue disease(s), Kawasaki syndrome, drug overdose,
drug or drug
withdrawal induced hyperthermia, alcohol/drug withdrawal, idiosyncratic drug
reaction, fever
of known or unknown or uncertain origin (non-limiting e.g. infectious
disease(s),
inflammation, immunological disease(s), non-infectious inflammatory disease(s)
{non-
limiting eg. systemic rheumatic and autoimmune diseases, vasculitis,
granulomatous diseases,
autoinflammatory syndromes}, tissue destruction, reaction to incompatible
blood product(s),
metabolic disorder(s), inherited metabolic disorder(s), cancer, neoplasm,
endogenous or
exogenous pyrogen(s), injury, head injury);
(v) disease/disorder/injury/pathology/surgery
treatable/ameliorated/prevented/combated/helped by conferring hypothermia in a
subject for
some medical or other purpose which can include slowing a chemical reaction(s)
rate in a
subject for therapeutic benefit, preventing/minimizing brain and/or tissue
damage, slowing
physiological/pathological processes (reaction rates are temperature
dependent) and so
"buying time" to get the subject to treatment for their emergency (e.g.
trauma/septic shock or
.. other medical emergency), slowing the progress of sepsis until a sufficient
concentration of a
working antibiotic(s) can be built up in the subject (furthermore hypothermia,
by slowing
sepsis progression, buys time to observe which antibiotic(s) can work,
yielding time to try
alternative further antibiotic option(s) if required), used soon after or just
before clinical/legal
death to preserve the subject's organs/tissues until the subject can be
frozen/cryogenically
frozen or the pathology that caused clinical/legal death (e.g. wound) can be
fixed and the
subject resuscitated, administered to a subject when a first responder (e.g.
ambulance crew)
deems the subject dead or unlikely to survive the journey to a medical
facility (e.g. hospital)
wherein this administration helps to preserve the subject which is helpful if
hospital staff
subsequently assess that they can, or might be able to, save the subject,
stabilizing
.. surgical/trauma/Emergency Room (ER) patients, deep hypothermic circulatory
arrest for
surgery (DHCA, non-limiting applications of DHCA include repairs of the aortic
arch, repairs
to head and neck great vessels, repair of large cerebral aneurysms, repair of
cerebral
arteriovenous malformations, pulmonary thromboendarterectomy, resection of
tumors that
have invaded the vena cava, brain tumor resection {wherein the anti-cancer
activity of a
compound(s) of this invention juxtaposes well}), Emergency Preservation and
Resuscitation
(EPR), hypothermia for a surgical purpose, protective hypothermia during
surgery and/or
surgery complication, hypothermia to slow/reduce blood loss, hypothermia for
neuro- and/or
cardio- and/or organ/tissue and/or life protection in a subject that has
trauma/brain
trauma/polytrauma/surgery/stroke/ischemic stroke/hemorrhagic stroke/cardiac
588
CA 3050553 2019-07-25
arrest/myocardial infarction/hypoxia/shock (including, without limitation, low
volume,
cardiogenic, obstructive, and distributive shock)/sepsis/septic shock/multiple
organ
dysfunction syndrome/systemic inflammatory response syndrome (SIRS)/organ
failure/cytokine storm/anaphylactic shock/seizure/disseminated intravascular
.. coagulation/blocked
airway/croup/rhabdomyolysis/[head/facial/spinal/chest/abdominal/ballistic/knife
injury/trauma], or some other medical
emergency/condition/disorder/disease/injury/operation,
hypothermia for cardiac and/or cardiovascular surgery and/or open heart
surgery and/or brain
surgery (neurosurgery) and/or surgery using total circulatory arrest and/or
surgery using
cardiopulmonary bypass, Emergency Preservation and Resuscitation (EPR),
preserving
detached body parts such as limbs and/or organs (for example during organ
storage/transport
and/or transplant, thus increasing the time window for transplantation of
organs to recipients;
compound(s) of this invention is administered to organ to be transplanted [by
administration
to donor and/or by administration to isolated organ] and/or to organ
recipient, optionally
during transplant operation), protective hypothermia, targeted temperature
management,
therapeutic hypothermia, hypothermia therapy for neonatal encephalopathy,
neonatal
hypoxia-ischemia, birth asphyxia, hypoxic-ischemic encephalopathy (HIE),
haemorrhage,
hypovolemia, exsanguination, suspended animation, decompression sickness, burn
injury(s)
including skin burn, inflammation, allergic reaction, anaphylaxis,
tissue/organ rejection,
hypoxia, hypoxemia, anoxemia, anoxia, anemia, hypervolemia, altitude sickness,
obstructed
airway, asthma attack, hypoxia in a body/tissue/organ, hypoglycemia,
reperfusion injury
(ischemia-reperfusion injury), upon release of a ligature or tourniquet,
uraemia, crush
syndrome, compartment syndrome, traumatic brain and/or spinal cord injury,
major trauma,
infection, bacterial and/or viral infection(s) (non-limiting e.g. meningitis),
sepsis, septic
shock, stroke, cerebrovascular disease, ischemic brain injury, ischemic
stroke, traumatic
injury, brain injury, spinal cord injury, cardiac arrest, heart failure,
congestive heart failure,
Dilated cardiomyopathy, valvular heart disease, pulmonary embolism, adrenal
crisis,
Addisonian crisis, hypertensive emergency, haemorrhagic (hypovolemic) shock,
cardiogenic
shock, neurogenic shock, hepatic encephalopathy, blood loss, ischemic
brain/heart/kidney/intestinal injury, neuroprotection and/or cardioprotection
and/or tissue
protection during/after a stroke and/or ischemia and/or cardiac arrest and/or
resuscitation
and/or a period(s) of poor blood flow anywhere in a subject;
(vi) poisoning by a toxic amount of a compound(s) in a subject (non-limiting
e.g.
carbon monoxide/methanol/heavy metal/ethylene glycol/pesticide poisoning,
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CA 3050553 2019-07-25
snake/spider/bee/insect/lizard venom, metabolic poison(s), nerve agent,
chemical weapon,
bacterial toxin(s) (e.g. food poisoning, Salmonella poisoning), endotoxemia,
eukaryote
produced toxin(s) e.g. (non-limiting) brevetoxin, drug(s)/substance(s)
overdose e.g. (non-
limiting) heroin, ethanol, a prescription medication(s), an over the counter
medication(s) such
as aspirin, paracetamol etc.; hypothermia is protective to toxic insult);
(vii) hypermetabolism (optionally because of one or more of, without
restriction,
traumatic brain injury, injury to the body, infection, sepsis, burn, multiple
trauma, fever,
long-bone fracture, hyperthyroidism, prolonged steroid therapy, surgery, bone
marrow
transplant, recovery from anorexia/bulimia), heat intolerence, insomnia, fatal
insomnia,
nervousness, Luft's disease, non-thyroidal hypermetabolism, thyrotoxicosis,
hyperthyroidism,
overactive thyroid, subclinical hyperthyroidism, too much thyroid hormone(s)
in the subject,
too much triiodothyronine (T3) and/or thyroxine (T4) in the subject,
hyperthyroxinemia
(including, without restriction, familial dysalbuminemic hyperthyroxinemia,
familial
euthyroid hyperthyroxinemia, thyroid hormone resistance syndrome), thyroid
storm,
hyperthyroidism caused by one or more of (without restriction) Graves'
disease, thyroiditis,
Hashimoto's thyroiditis, subacute thyroiditis, postpartum thyroiditis, lumps
(nodules) on the
thyroid, enlarged thyroid gland (goitre), simple goitre, multinodular goiter,
toxic multinodular
goiter, toxic adenoma, toxic thyroid adenoma, inflammation of the thyroid,
hyperplasia of
thyroid, metastatic thyroid cancer, thyroid tumour, thyroid cancer (including,
without
restriction, papillary carcinoma, follicular carcinoma, medullary thyroid
carcinoma,
anaplastic thyroid carcinoma), eating too much iodine, goitrogen ingestion,
consumption of
ground beef contaminated with thyroid tissue ("hamburger hyperthyroidism"),
too much
synthetic thyroid hormone in the subject, pituitary adenoma, drug induced,
Amiodarone drug
induced, struma ovarii, Jod-Basedow syndrome, nonautoimmune autosomal dominant
hyperthyroidism;
(viii) low or less than desired metabolic/bioenergetic efficiency in a
subject, or low or
less than desired physical or mental performance (e.g. memory, IQ), or low or
less than
desired body weight, or fatigue/tiredness/weakness/exhaustion;
(ix) accelerated aging disease or progeroid syndrome including, without
restriction,
Werner syndrome, Bloom syndrome, De Barsy syndrome, Rothmund¨Thomson syndrome,
Cockayne syndrome, xeroderma pigmentosum, trichothiodystrophy, combined
xeroderma
pigmentosum-Cockayne syndrome, restrictive dermopathy, Wiedemann¨Rautenstrauch
syndrome, Hutchinson¨Gilford progeria syndrome (progeria), Ataxia
telangiectasia-like
disorder 2, XFE progeroid syndrome, Muscular dystrophy, Muscular Dystrophy
(Becker's,
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CA 3050553 2019-07-25
Duchenne, Limb-Girdle), Yamamoto's Muscular Dystrophy, Mandibuloacral
dysplasia,
Dilated cardiomyopathy, GAPO syndrome, Cutis laxia, Ehlers-Danlos syndrom,
Lenz-
Majewski hyperostatic dwarfism, SHORT syndrome, Progressive external
opthalmoplegia,
Nester-Guillermo progeria syndrome, MDPL syndrome, Dyskeratosis congenital,
Down
syndrome;
(x) disease or disorder of aging (incidence/severity increases with increased
age/senescence) and/or unwanted/undesirable aspect(s) of aging and/or a
disease/disorder
associated with elevated reactive oxygen species including age-associated
decline, aging
frailty, frailty syndrome, sarcopenia, muscle weakness, osteoporosis,
cognitive decline,
cognitive defecit, mild cognitive impairment, degenerative diseases,
neurodegenerative
diseases, motor-associated neurodegenerative diseases, motor neuron disease,
amyotrophic
lateral sclerosis (ALS), primary lateral sclerosis, progressive muscular
atrophy, progressive
bulbar palsy, progressive supranuclear palsy, pseudobulbar palsy, hereditary
spastic
paraplegia, Parkinson's disease, Multiple System Atrophy (MSA), Progressive
Supranuclear
Palsy (PSP), essential tremor, resting tremor, Alzheimer's disease,
Huntington's disease,
spinocerebellar ataxias, Friedreich's ataxia, dementia, frontotemporal
dementia, chronic
traumatic encephalopathy, memory loss, aged cognition, age/aging related
cognitive
decline/impairment, Batten disease, polyglutamine diseases, osteoporosis,
atherosclerosis,
cardiovascular disease, myocardial infarction, cerebrovascular disease,
stroke, heart failure,
heart failure with preserved ejection fraction, idiopathic pulmonary fibrosis,
fibrotic disease,
pulmonary disease, coronary artery disease, hypercholesterolemia, obesity,
liver disease, fatty
liver disease, lysosomal storage disease, amyloidosis, systemic sclerosis,
kidney disease,
hepatic cirrhosis, immunosenscence, clonal hematopoiesis, chronic obstructive
pulmonary
disease (COPD), hypertension, hypercholesterolemia, age-related thymic
atrophy, arthritis,
osteoarthritis, arthritis (Osteo-and Rheumatoid), Juvenile Rheumatoid
Arthritis (JRA),
Degenerative Disc Disease, Tendinopathy, Androgenetic Alopecia, male-pattern
baldness,
Idiopathic Pulmonary Fibrosis, systemic sclerosis, Chronic Obstructive
Pulmonary Disease,
Psoriasis, age-related loss of cardiac/pulmonary/cognitive/vision function,
diabetes, type 2
diabetes, andropause, glaucoma, retinal degeneration, sarcopenia, cachexia,
age-related
cachexia and/or sarcopenia, age-related macular degeneration (AMD,
early/intermediate/late), neovascular/wet AMD, dry AMD, Geographic atrophy
(GA), wet
and dry AMD in the same eye(s), Stargardt's macular degeneration, Best
vitelliform macular
dystrophy, diabetic retinopathy, proliferative diabetic retinopathy, diabetic
macular edema,
age/aging-related eye disease, ophthalmological disease/disorder, ocular
disease, vision loss,
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CA 3050553 2019-07-25
progressive vision impairment, myopia (short-sightedness), degenerative
myopia, hyperopia
(far-sightedness), accommodative dysfunction, glaucoma, cataract formation,
retinal
degeneration, progressive retinal degeneration, retinitis pigmentosa, leber
hereditary optic
neuropathy, Fuchs spot, Best's disease, Sorsby's fundus dystrophy, hearing
loss (e.g. age-
related), presbycusis, tinnitus, naive T cell shortage, movement disability,
nonalcoholic fatty
liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), immunosenescence,
respiratory/urinary tract infection (RTI/UTI) especially in older/aged/elderly
subjects, cancer;
(xi) aging and/or one or more signs of aging, wherein one or more of these
compounds slow/delay/reduce/treat/prevent/stop/reverse aging, and/or extend
lifespan and/or
.. healthspan, and/or treat or delay the onset of geriatric aging of the
human/animal body,
tissue(s), or organ(s), and/or treat or delay the onset of an age-associated
phenotype in a
cell(s)/organism(s), and/or prolong fertility e.g. female fertility, delay
menopause;
(xii) skin aging and/or damage (including sun damage) and/or scalp and/or hair
aging
and/or hair greying and/or hair loss;
(xiii) insomnia, fatal insomnia, sleep onset latency, delayed sleep phase
disorder,
exploding head syndrome, parasomnia, sleep-maintenance insomnia, sleep
disorder, too
much/inappropriate/undesired signals/activity/electrical activity in the
nervous system,
hyperactivity, hypersensitivity, Premature ejaculation, hyperreflexia,
Autonomic dysreflexia
(AD), Hyperventilation syndrome, brain hyperactivity, overly sensitive sensory
system,
pathological crying and/or laughing, Pseudobulbar affect (PBA, emotional
lability),
photophobia, phonophobia, temperature-sensitive, pressure-sensitive, brain
hyperexcitability,
overstimulation, intrusive thought(s), Perseveration, sensory overload,
disorganized thinking,
fantasy prone personality, malapdative daydreaming, dissociation, hyperkinetic
disorder,
agitation, Psychomotor agitation, restlessness, difficulty controlling
behaviour, disruptive
behaviour disorder, Emotional and behavioral disorder, pervasive developmental
disorder,
Overactive disorder associated with mental retardation and stereotyped
movements, attention-
deficit disorder, Attention Deficit Hyperactivity Disorder (ADHD), adult
attention-deficit
hyperactivity disorder, severe behavioral problem(s) in children (e.g., to
illustrate and not
restrict, combativeness and/or explosive hyperexcitable behavior {out of
proportion to
immediate provocation[s]}, hyperactive children who show excessive motor
activity with
accompanying conduct disorders consisting of one or more of: impulsivity,
difficulty
sustaining attention, aggressivity, mood lability, poor frustration
tolerance), Premenstrual
dysphoric disorder (PMDD), premenstrual syndrome (PMS), impulsiveness,
impulsivity,
impulse control disorder, lack of self-control, hysteria, histrionic
personality disorder,
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CA 3050553 2019-07-25
attention difficulty, inattention, poor attention control, anxiety, paranoid
anxiety, Paranoid
personality disorder, distress, dysphoria, Adjustment disorder, separation
anxiety, anxiety
disorder, depressive anxiety, agitated depression, treatment-resistant
depression, Generalized
anxiety disorder, social anxiety disorder, stranger anxiety, separation
anxiety (e.g. in dogs left
at home), separation anxiety disorder, Mixed anxiety-depressive disorder,
depression (all
forms, all severities), restlessness/apprehension/anxiety before surgery.
hypochondria, panic
disorder, panic attack, emotional outburst, emotional instability,
Intermittent explosive
disorder, unreasonable/unwarranted anger/aggression, hyper-aggression,
hostility, rage, poor
temper control, self-hatred, poor attentional control, worry, irritability,
neuroses, somatization
disorder, somatic symptom disorder, pain disorder, psychological pain,
psychogenic pain,
psychogenic facial pain, Atypical odontalgia (AO), burning mouth syndrome,
throbbing,
toothache/pulpitis/dental pain, chronic lower back pain, negative emotion,
persistent/enduring
negative emotion, body dysmorphic disorder, factitious disorder, illness
anxiety disorder,
unwarrented fight-or-flight response, stress, emotional stress, emotional
dysregulation,
distress, psychological stress, acute stress, chronic stress, acute stress
reaction, combat stress
reaction, traumatic grief, grief, grief after death of loved one, Prolonged
grief disorder (PGD),
heartbreak, guilt, shame, remorse, emotional pain, Algopsychalia, psychalgia,
suffering,
emotional trauma, psychological trauma, broken heart, Post Traumatic Stress
Disorder
(PTSD), Complex post-traumatic stress disorder (C-PTSD), hypervigilance,
sympathetic
hyperactivity, inability or impaired ability to relax, flashbacks, dysphoric
hyperarousal,
agoraphobia, insomnia, fatal insomnia, mood disorder, irrational/unwarrented
fear/terror,
phobia, social phobia, Cancerophobia, thunderstorm/firework phobia,
hypersexuality,
hypersexual disorder, depression, clinical depression, unipolar depression,
bipolar disorder,
Bipolar I, Bipolar II, Bipolar disorder not otherwise specified (Bipolar NOS),
cyclothymia,
cyclothymic disorder, mixed affective state, atypical depression, melancholic
depression,
postpartum depression, double depression, seasonal affective disorder, mania,
manic episode,
hypomania, increase in energy of psychomotor activity, delirium, excited
delirium, major
depressive disorder, minor depressive disorder, recurrent brief depression,
Depressive
Disorder Not Otherwise Specified (DD-NOS), major depressive episode,
persistent
depressive disorder (PDD), dysthymia, dysthymic disorder, absence of euthymia,
manic
thoughts, racing thoughts, thought disorder, disordered thinking, reduced
ability to plan and
execute tasks, paranoia, hallucination (including, without limitation, visual,
auditory,
olfactory, gustatory, tactile, proprioceptive, equilibrioceptive, nociceptive,
thermoceptive,
chronoceptive), Charles Bonnet syndrome, delusion, persecutory delusion,
hearing voices,
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CA 3050553 2019-07-25
homicidal/criminal ideation/tendency/thoughts, suicidal
ideation/tendancy/thoughts, self-
injury, non-suicidal self-injury, violence, attacking others, negative mood
swing, personality
disorder, Borderline personality disorder, Narcissistic personality disorder,
malignant
narcissism, dissociative disorder, dissociative identity disorder (DID),
Psychosis, acute
psychosis, chronic psychosis, psychosis spectrum disorder, manifestations of
psychotic
disorders, behavioral complications of mental retardation, stimulant
psychosis, psychotic
depression, hallucinogen persisting perception disorder, Psychoactive
substance-related
disorder, amphetamine-induced psychosis, brief psychotic disorder, Brief
reactive psychosis,
Menstrual psychosis, postpartum psychosis, Psychotic disorder, Psychopathy,
chronic
hallucinatory psychosis, manifestation(s) of psychotic disorder,
neurotic/reactive/endogenous/involutional/psychotic depression/depressive
disorder
(optionally accompanied by anxiety or agitation), depressive neurosis,
delusional depression,
psychotic aggression, psychiatric symptoms of dementia, AIDS delirium,
Supersensitivity
psychosis, Tardive psychosis, Tardive dysmentia, Depersonalization disorder,
out-of-body
experience, Sociopathy, Schizophrenia, Paranoid schizophrenia, disorganized-
type
schizophrenia, simple-type schizophrenia, pseudoneurotic schizophrenia,
prodromal
schizophrenia, schizoaffective disorder, bipolar type schizoaffective
disorder, depressive type
schizoaffective disorder, schizoaffective psychosis, Schizotypal personality
disorder,
schizophreniform disorder, Delusional parasitosis, formication, paresthesias,
Acroparesthesia,
tinnitus, delusional disorder, delusional jealousy, inappropriate behaviour,
behavioural
disorder, antisocial personality disorder, Oppositional defiant disorder
(ODD), conduct
disorder (CD), Disruptive mood dysregulation disorder (DMDD), sadistic
personality
disorder, poor inhibitory control, kleptomania, Pyromania, trichotillomania,
dermatillomania,
pathological/problem gambling, dyskinesia, tardive dyskinesia, paroxysmal
dyskinesia,
Paroxysmal kinesigenic dyskinesia, Paroxysmal nonkinesigenic dyskinesia,
Paroxysmal
exercise-induced dystonia, Hemiballismus, tic disorder, tremor, clonus,
Tourette's syndrome,
coprolalia, copropraxia, Echophenomena, Echopraxia, Echolalia, Palilalia,
stuttering/stammering, stereotypy, punding, Self-stimulatory behaviour
(stimming),
Stereotypic movement disorder (SMD), synesthesia, obsession,
Obsessive¨compulsive
disorder (OCD), obsessive¨compulsive personality disorder, anankastic
personality disorder,
relationship obsessive¨compulsive disorder (ROCD), Scrupulosity, Primarily
obsessional
obsessive compulsive disorder, sexual obsession, Akathisia (including, without
limitation,
chronic, acute, Pseudoakathisia, Tardive akathisia, withdrawal or "rebound"
akathisia),
Restless legs syndrome, motor restlessness, periodic limb movement disorder
(PLMD),
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CA 3050553 2019-07-25
periodic limb movements in sleep (PLMS), Sydenham's chorea, chorea, dystonia,
Hypnic
jerk, twitching, spasms, Tetanus, tetanus muscle spasms, tetanized state,
Myoclonus,
myoclonic seizure, Action myoclonus, Palatal myoclonus, Middle ear myoclonus,
Spinal
myoclonus, Stimulus-sensitive myoclonus, Sleep myoclonus, Cortical reflex
myoclonus,
Essential myoclonus, myoclonic epilepsy, Juvenile myoclonic epilepsy,
Progressive
myoclonus epilepsy (PME, including, without limitation, Dentatorubral-
pallidoluysian
atrophy, Unverricht-Lundborg disease, MERRF syndrome, Lafora disease),
Reticular reflex
myoclonus, Myoclonic epilepsy, diaphragmatic flutter, automatism, status
epilepticus,
Epilepsia partialis continua, Complex partial status epilepticus, epilepsy,
epileptic seizure,
simple partial seizure, complex partial seizure, generalized epilepsy,
generalized seizure
(including, without limitation, tonic-clonic, tonic, clonic, myoclonic,
absence (including
typical absence and atypical absence), atonic seizure), focal epilepsy, focal
seizure,
focal/partial seizure (including, without limitation, Simple partial seizure
and Complex partial
seizure), focal aware seizure, focal impaired awareness seizure, generalised
epilepsy,
temporal lobe epilepsy (including, without restriction, mesial temporal lobe
epilepsy
{MTLE} and lateral temporal lobe epilepsy ILTLED, Frontal lobe epilepsy,
Rolandic
epilepsy, Nocturnal epilepsy, Nocturnal frontal lobe epilepsy, Autosomal
dominant nocturnal
frontal lobe epilepsy (ADNFLE), Generalized epilepsy with febrile seizures
plus (GEFS+),
Panayiotopoulos syndrome, epileptic fit, reflex epilepsy, reflex seizure,
absence seizure
(including, without limitation, childhood absence epilepsy, epilepsy with
myoclonic
absences, juvenile absence epilepsy, juvenile myoclonic epilepsy, Jeavons
syndrome {eyelid
myoclonia with absences), genetic generalised epilepsy with phantom absences),
complex
partial seizure, atonic seizure, generalized tonic-clonic seizure,
tonic¨clonic seizure, extrinsic
stimulus epilepsy, intrinsic stimulus epilepsy, photosensitive epilepsy,
musicogenic epilepsy,
thinking epilepsy, eating epilepsy, seizure(s), Febrile seizure, nerve agent
induced seizure,
Dravet syndrome (sometimes modest hyperthermic stressors like physical
exertion or a hot
bath can provoke seizures in affected individuals), acute symptomatic seizure,
seizure-related
disorder, drug related seizure, paroxysmal depolarizing shift, Ohtahara
syndrome, Epilepsy in
females with mental retardation, Rasmussen's encephalitis, Epilepsy syndrome,
benign
rolandic epilepsy, childhood absence epilepsy, absence epilepsy, juvenile
myoclonic
epilepsy, epileptic encephalopathies, Lennox¨Gastaut syndrome, West syndrome
(Epileptic
spasms), Doose syndrome (Myoclonic astatic epilepsy, MAE), Lennox-Gestaut
syndrome,
pseudo-Lennox Gastaut syndrome (atypical benign partial epilepsy), Benign
familial neonatal
epilepsy, Benign occipital epilepsy of childhood, familial neonatal
convulsions, Febrile
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CA 3050553 2019-07-25
infection-related epilepsy syndrome, interictal dysphoric disorder, euphoria
sclerotic,
psychogenic non-epileptic seizure, non-epileptic seizure, gelastic seizure,
convulsion(s),
migraine, status migrainosus, tension headache, headache, Hypnic headache,
hiccups,
intractable hiccups, thumps in equines, Postural orthostatic tachycardia
syndrome (POTS),
Pheochromocytoma, agony, pain, chronic pain, acute pain, pain due to
disease/injury,
neuropathic pain, fibromyalgia, postherpetic neuralgia, phantom pain, referred
pain, back
pain, lower back pain, pelvic pain, cancer associated pain, chemotherapy
associated pain,
Diabetic neuropathy, small fiber peripheral neuropathy, Mononeuritis
multiplex,
Wartenberg's migratory sensory neuropathy, Breakthrough pain, idiopathic pain,
polyneuropathy, Neuritis, Mononeuropathy, Polyradiculoneuropathy, Radial
neuropathy,
neuropathy, visceral pain, Burning feet syndrome, Tarsal tunnel syndrome,
Carpal tunnel
syndrome, Guyon's canal syndrome, Cubital Tunnel Syndrome, Repetitive strain
injury,
Sciatica, Neurofibromatosis, Ulnar nerve entrapment, Erythromelalgia,
Paroxysmal extreme
pain disorder, Proctalgia fugax, dysesthesia, scalp dysesthesia, dysesthetic
burning,
hyperesthesia, hyperalgesia, Opioid-induced hyperalgesia, hyperpathia,
allodynia, pain
response from stimuli which do not normally provoke pain, Complex regional
pain syndrome
(said to be most painful condition known to man), Radiculopathy, neuralgia
(including,
without restriction, intercostal neuralgia, trigeminal neuralgia, atypical
trigeminal neuralgia,
glossopharyngeal neuralgia, occipital neuralgia, postherpetic neuralgia),
ciguatera poisoning,
irritable bowel syndrome (IBS), interstitial cystitis (IC), temporomandibular
joint disorder,
acute intermittent porphyria, Porphyria, Acute porphyria (including, without
limitation, acute
intermittent porphyria {AIP}, variegate porphyria {VP}, aminolevulinic acid
dehydratase
deficiency porphyria {ALAD}, hereditary coproporphyria {HCP}, drug induced),
Chronic
porphyria (including, without limitation, X-linked dominant protoporphyria
{XLDPP},
congenital erythropoietic porphyria {CEP}, porphyria cutanea tarda {PCT}, and
erythropoietic protoporphyria {EPP}), cutaneous porphyria, Porphyria cutanea
tarda, allergy,
allergic reaction, anaphylaxis, anaphylactic shock, hives, asthma, allergic
rhinitis, rhinitis,
urticaria, contact dermatitis, cough, sore throat, esophageal reflux disease,
heartburn, chest
pain, Esophageal motility disorder, Nutcracker esophagus, diseases involving
gastrointestinal
motility, Peptic ulcer disease, esophageal spasm, angina, itchiness, Pruritus,
severe pruritus,
Prurigo, Pruritic skin condition, chronic itch, eczema, pruritus in eczema,
neuropathic itch,
neurogenic itch, itchiness due to atopic dermatitis and/or lichen simplex
chronicus, peripheral
sensitization, central sensitization, sensory perception of absent stimuli,
too much sensory
stimulation, sensory stimulation brings discomfort, Neuromyotonia, Peripheral
nerve
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CA 3050553 2019-07-25
hyperexcitability, Morvan's syndrome, Benign fasciculation syndrome, Cramp
fasciculation
syndrome, hyperhidrosis, undifferentiated somatoform disorder, somatoform
disorder,
somatic symptom disorder, conversion disorder, functional neurological symptom
disorder,
severe nausea and/or vomiting, severe nausea/emesis, Chemotherapy-induced
peripheral
neuropathy, chemotherapy-induced nausea and vomiting (CINV), radiation therapy-
induced
nausea and vomiting (RINV), postnarcotic nausea, hyperemesis gravidarum,
morning
sickness, Cyclic vomiting syndrome, retching/dry heaving, Urinary
incontinence, enuresis,
nocturnal enuresis, tail chasing, reduce urine spraying/marking behavior,
benzodiazepine
withdrawal syndrome, barbituate withdrawal syndrome, Neuroleptic
discontinuation
syndrome, drug withdrawal discomfort/pain/symptoms, drug use disorder, alcohol
use
disorder, opoid use disorder, amphetamine use disorder, cocaine use disorder,
alcohol
withdrawal syndrome/symptoms, delirium tremens, opoid withdrawal
sydrome/symptoms,
drug craving, drug addiction, drug dependence, polysubstance dependence, drug
overdose,
smoking, tobacco (nicotine) addition, tobacco (nicotine) withdrawal symptoms,
alcoholism,
addiction, opoid addiction, cocaine/crack addiction, addictive behaviour,
addictive
personality, behavioural addiction, internet/computer/computer game/social
media/media
addiction, exercise addiction, compulsive behaviour (e.g. {non-limiting}
checking, counting,
washing, repeating), anti-social behaviour, criminality, sexual compulsion,
impulsive sexual
behaviour, compulsive buying, gambling addiction, sex related addiction,
sexual urge,
hunger, eating desire/compulsion, eating disorder, polyphagia, overeating,
binge eating
disorder, compulsive overeating, insatiable/excessive appetite, bulimia
nervosa, anorexia
nervosa, substance abuse, substance-induced delirium, substance-induced
psychosis,
substance-induced mood disorder, drug overdose, vertigo, motion sickness,
seasickness,
mental/nervous breakdown, Autism spectrum disorder, neurological disorder,
cognitive
disorder, mental disorder, mental health disorder, mental health condition
involving impaired
or altered neural plasticity, mood disorder, mental disorder disclosed in
Diagnostic and
Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) or a later
edition, a
mental/behavioural disorder disclosed by the International Classification of
Diseases (LCD) in
ICD-10 Chapter V: Mental and behavioural disorders (World Health Organisation,
WHO); or
(xiv) diseases or disorders or conditions or pathologies or
unwanted/undesirable
effects/actions/behaviour treatable/ameliorated/prevented/combated, in
totality or in part (e.g.
along with surgery), by anaesthesia, pre-anaesthesia, post-anaesthesia,
hypoesthesia,
hypoactivity, sedation, coma, tranquilization, behavioural submission, muscle
relaxation,
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CA 3050553 2019-07-25
hibernation, artificial hibernation, torpor, synthetic torpor, suspended
animation (e.g. used
during spaceflight because e.g. reduces ionizing radiation damage to subject);
(xv) hyperproliferative/hyperplasia disorder, non-cancerous proliferative
disorder,
hyperproliferative autoimmune disorder, hyperplasia, epidermal hyperplasia,
dysplasia (e.g.
epithelial dysplasia), nodule(s), wart(s), papilloma(s), squamous cell
papilloma, genital
wart(s), condyloma(s), condyloma acuminatum, cyst(s), polyp(s) {including,
without
restriction, digestive, colorectal, endometrial, cervical, nasal, laryngeal,
inflammatory fibroid
polyp[s])}, inherited/hereditary (including, without restriction, Familial
adenomatous
polyposis, Peutz¨Jeghers syndrome, Turcot syndrome, Juvenile polyposis
syndrome, Cowden
disease, Bannayan¨Riley¨Ruvalcaba syndrome {Bannayan-Zonana syndrome},
Gardner's
syndrome) and non-inherited (non-restrictive e.g. Cronkhite¨Canada syndrome)
polyposis
syndrome, benign tumour, adenoma, organ enlargement by hyperplasia, Cushing's
disease
(enlarged adrenal cortex by hyperplasia), congenital adrenal hyperplasia,
hyperplasia of
breast, atypical ductal hyperplasia, intraductal papillomatosis,
fibroadenomas, fibrocystic
changes, hemihyperplasia, focal epithelial hyperplasia, sebaceous hyperplasia,
sebaceous
adenoma, intimal hyperplasia, unwanted/undesirable smooth muscle cell
proliferation,
smooth muscle cell hyperplasia, intimal smooth muscle cell hyperplasia,
neointimal
hyperplasia, proliferative vascular disorders, stenosis, stenosis because of
cellular
proliferation, vaginal stenosis, stenosis in a blood vessel, lessoned patency
of a blood vessel,
stenosis in a blood vessel because of cellular proliferation, vascular
occlusion, restenosis,
restenosis in a blood vessel that has been implanted with a stent, in-stent
restenosis, post-
angioplasty restenosis, systemic sclerosis, cirrhosis of the liver, adult
respiratory distress
syndrome, idiopathic cardiomyopathy, lupus erythematosus, retinopathy (e.g.
diabetic
retinopathy and/or other retinopathy[y/ies]), cardiac hyperplasia, fibrosis,
pulmonary fibrosis,
idiopathic pulmonary fibrosis, fibromatosis, neurofibromatosis, renal
interstitial fibrosis,
Cowden syndrome, hamartoma(s), choristoma(s), hemangioma(s), lymphangioma(s),
rhabdomyoma(s), lymphangiomatosis, cystic hygroma, trichilemmoma, sarcoidosis,
neurosarcoidosis, aggressive fibromatosis, desmoid tumour(s),
unwanted/undesirable skin
cell proliferation, hyperproliferative skin disorder, psoriasis (including,
without restriction,
plaque, guttate, inverse, pustular, napkin, seborrheic-like, nail, scalp and
erythrodermic
psoriasis), psoriatic arthritis, dactylitis, seborrhoeic dermatitis, dandruff,
eczema, atopic
dermatitis, rosacea, reactive arthritis (Reiter's syndrome), pityriasis rubra
pilaris,
hyperproliferative variants of disorders of keratinization (e.g., without
restriction, actinic
keratosis, senile keratosis, keratosis pilaris, seborrheic keratosis),
scleroderma, benign
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CA 3050553 2019-07-25
prostatic hyperplasia, prostate enlargement, endometrial hyperplasia, atypical
endometrial
hyperplasia, benign endometrial hyperplasia, adenomyosis, atypical polypoid
adenomyoma,
endometriosis, endometriosis of ovary (endometrioma), endometrial polyp(s),
polycystic
ovary syndrome, ovarian cyst(s), cervical polyp(s), uterine fibroid(s),
uterine hyperplasia;
(xvi) Tumour Associated Macrophages (TAMs) or any macrophage associated
disease or disorder such as, without limitation, Macrophage Activation
Syndrome (MAS),
HIV, AIDS, HIV-associated neurocognitive disorders (HAND), AIDS dementia, HIV
peripheral neuropathy, HIV associated cancers, AIDS-defining cancers, non-AIDS
defining
cancers, any disease in which the pathogen(s) hides from the immune system in
macrophages
including, without limitation, Mycobacterium tuberculosis (causes
tuberculosis), Leishmania
parasite (causes Leishmaniasis), Chikungunya virus (causes Chikungunya),
Legionella
pneumophila (causes Legionnaires' disease), adenoviruses, T. whipplei (causes
Whipple's
Disease), Brucella spp. (causes brucellosis), Staphylococcus aureus, Ebola
virus, Hepatitis B
virus, Hepatitis C virus, influenza virus strains, dengue virus and antibiotic
resistant bacteria,
any disease or condition in which activated macrophages are unwanted or
undesirable;
(xvii) virus/pathogen neuroinvasion via macrophage(s), as used for non-
limiting
example by HIV, Heptatitis C virus and SARS coronavirus;
(xviii) neurocognitive or neurodegenerative diseases/disorders, for non-
limiting
example those caused by a virus;
(xix) virus/pathogen transmission from mother to fetus/baby via macrophage(s)
as
used for non-limiting example by zika (via Hofbauer cells) and HIV
(macrophages in breast
milk);
(xx) acute or chronic or systemic inflammation or any inflammatory
disease/disorder/syndrome or any autoinflammatory disease/disorder/syndrome or
any
autoimmune disease/disorder/syndrome;
(xxi) acute inflammation, chronic inflammation, systemic inflammation,
inflammation
because of infection or foreign bodies or injury or chemical or toxin or drug
or stress or
frostbite or burn or ionising radiation or surgery, inflammatory
diseases/disorders/syndromes,
Macrophage Activation Syndrome (MAS), autoinflammatory
diseases/disorders/syndromes,
age-related chronic inflammatory diseases ("inflammaging"), autoimmune
diseases/disorders/syndromes, diseases/disorders of the innate immune system,
sore throat,
sore throat associated with cold or flu or fever, high-intensity exercise
associated
inflammation, ulcerative colitis, inflammatory bowel disease (IBD), irritable
bowel syndrome
(IBS), rheumatoid arthritis, osteoarthritis, inflammatory osteoarthritis,
psoriatic arthritis,
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atopic dermatitis, allergic airway inflammation, asthma, inflammation
associated depression,
neuroinflammation, neuropathic pain, exercise-induced acute inflammation,
atherosclerosis,
allergy, hay fever, anaphylaxis, inflammatory myopathies, drug-induced
inflammation,
systemic inflammatory response syndrome, sepsis-related multiple organ
dysfunction/multiple organ failure, microbial infection, acute
brain/lung/hepatic/renal
injuries, lung inflammation, acute lung injury (ARDS), acne vulgaris, celiac
disease, celiac
sprue, chronic prostatitis, colitis, autoimmune hemolytic anemia,
diverticulitis,
glomerulonephritis, proliferative glomerulonephritis, membranous nephropathy,
minimal
change nephrotic syndrome, hidradenitis suppurativa, hypersensitivities,
interstitial cystitis,
Mast Cell Activation Syndrome, mastocytosis, otitis, pelvic inflammatory
disease (PID),
endometritis, reperfusion injury, rheumatic fever, rhinitis, sarcoidosis,
transplant rejection,
parasitosis, eosinophilia, type III hypersensitivity, ischaemia, chronic
peptic ulcer,
tuberculosis, Crohn's disease, hepatitis, chronic active hepatitis, immune
hepatitis, alcoholic
hepatitis, chronic viral hepatitis, ankylosing spondylitis, diverticulitis,
fibromyalgia, systemic
lupus erythematous (SLE), Alzheimer's disease, Parkinson's disease,
neurodegenerative
disease, cardiovascular disease, chronic obstructive pulmonary disease,
bronchitis, acute
bronchitis, bronchiectasis, bronchopneumonia, obliterative bronchiolitis,
appendicitis, acute
appendicitis, bursitis, cystitis, dermatitis, encephalitis, HIV encephalitis,
gingivitis,
meningitis, infective meningitis, myelitis, nephritis, neuritis,
periodontitis, chronic
periodontitis, phlebitis, prostatitis, RSD/CRPS, rhinitis, sinusitis, chronic
sinusitis, tendonitis,
testiculitis, tonsillitis, urethritis, vasculitis, respiratory
bronchiolitis¨associated interstitial
lung disease and desquamative interstitial pneumonia, pneumonia, interstitial
lung disease,
Lofgren syndrome, Heerfordt syndrome, monocytosis, liver fibrosis,
steatohepatitis,
nonalcoholic steatohepatitis, silicosis, histiocytoses, Langerhans' cell
histiocytosis,
haemophagocytic lymphohistiocytosis, pulmonary langerhans cell histiocytosis,
obesity, type
II diabetes, gout, pseudogout, organ transplant rejection, epidermal
hyperplasia, chronic
fatigue syndrome, graft versus host disease (GVHD), lymphadenopathy,
rheumatoid arthritis
(RA), osteoarthritis (OA), inflammatory osteoarthritis, lupus, multiple
sclerosis (MS),
myocarditis, uveitis, CNS disease(s), inflammation aspect to a CNS disease(s),
hypothalamic
inflammation, dementia, glaucoma, amyloid related/driven disease, lipid
storage disease(s),
fibrosis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic
steatohepatitis (NASH),
cirrhosis, cirrhosis of the liver, alcoholic cirrhosis, nephropath(y/ies),
lupus nephritis,
immune nephritis, fibrotic disorder(s), cardiovascular disease, heart disease,
atherosclerosis,
vulnerable plaque, plaque formation, lipid containing macrophage related
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disease(s)/disorder(s)/malad(y/ies), macrophage foam cells, diabetes, type I
diabetes, type 2
diabetes, insulin resistance, macrophage aspect to insulin resistance,
obesity, obesity
associated inflammation, macrophage accumulation/large numbers of macrophages
in
adipose tissue (e.g. associated with obesity), granuloma(s), granulomatous
diseases,
sarcoidosis (including, without limitation, Annular sarcoidosis, Erythrodermic
sarcoidosis,
Ichthyosiform sarcoidosis, Hypopigmented sarcoidosis, Lofgren syndrome, Lupus
pernio,
Morpheaform sarcoidosis, Mucosal sarcoidosis, Neurosarcoidosis, Papular
sarcoid, Scar
sarcoid, Subcutaneous sarcoidosis, Systemic sarcoidosis, Ulcerative
sarcoidos),
neurosarcoidosis, pulmonary sarcoidosis, interstitial lung disease, pulmonary
fibrosis,
pulmonary tuberculosis, immune reconstitution syndrome of HIV,
Jarisch¨Herxheimer
reaction, sepsis, Paget's disease of bone, osteolysis, monocytosis,
histiocytosis, X-type
histiocytoses, non-X histiocytoses, Langerhans cell histiocytosis, non-
Langerhans-cell
histiocytosis, malignant histiocytosis, malignant histiocytic disorders,
histiocytomas,
histiocytic lymphoma, hemophagocytic syndrome, hemophagocytic
lymphohistiocytosis,
lymphohistiocytosis, diffuse histiocytic sarcoma, Rosai¨Dorfman disease,
gliosis, Bergmann
gliosis, Chronic Obstructive Pulmonary Disease (COPD), chronic inflammatory
lung disease,
familial mediterranean fever (FMF), TNF receptor-associated periodic syndrome
(TRAPS),
Hyperimmunoglobulinemia D with recurrent fever syndrome (HIDS), cryopyrin
associated
periodic syndrome (CAPS), Blau syndrome, Majeed syndrome, deficiency of
interleukin-1
receptor antagonist (DIRA), mevalonate kinase deficiency, pyogenic-arthritis-
pyoderma
gangrenosum and acne syndrome (PAPA), periodic fever aphthous stomatitis
pharyngitis
adenitis (PFAPA) syndrome, Behcet's disease, Still's disease, Crohn's disease,
Schnitzler's
syndrome, Sweet's syndrome, NLRP12-associated autoinflammatory disorders,
deficiency of
interleukin-1 receptor antagonist (DIRA), pyoderma gangrenosum, cystic acne,
aseptic
.. arthritis, periodic Fever Associated with mevalonate kinase deficiency
(hyperimmunoglobulin D Syndrome), Pyogenic Arthritis Pyoderma Gangrenosum Acne
(PAPA) syndrome, Periodic Fever Aphthous Stomatitis, Pharyngitis and
Adenopathy
(PFAPA) syndrome, Adult-Onset Still's Disease (AOSD), Systemic Juvenile
Idiopathic
Arthritis (sJIA), Chronic Recurrent Multifocal Osteomyelitis (CRMO), Synovitis
Acne
Pustulosis Hyperostosis Osteitis (SAPHO) syndrome, Cryopyrin associated
Periodic
Syndrome (CAPS), Familial cold auto inflammatory syndrome (FCAS), Muckle-Wells
syndrome (MWS), Familial cold urticarial, Neonatal onset multisystemic
inflammatory
disorder (NOMID), hereditary Periodic Fever Syndromes, Periodic Fever
Syndromes,
systemic autoinflammatory diseases, Addison's disease, Agammaglobulinemia,
Alopecia
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areata, Amyloidosis, Ankylosing spondylitis, Anti-GBM/Anti-TBM nephritis,
Antiphospholipid syndrome, autoimmune angioedema, autoimmune dysautonomia,
autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune inner ear
disease
(AIED), autoimmune myocarditis, autoimmune pancreatitis, autoimmune
retinopathy,
autoimmune urticaria, axonal & neuronal neuropathy (AMAN), Balo disease,
Behcet's
disease, benign mucosal pemphigoid, Bullous pemphigoid, Castleman disease
(CD), Celiac
disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy
(CIDP),
chronic recurrent multifocal osteomyelitis (CRMO), Churg-Strauss, Cicatricial
pemphigoid,
Cogan's syndrome, cold agglutinin disease, congenital heart block, coxsackie
myocarditis,
CREST syndrome, Berger's disease, dermatitis herpetiformis, dermatomyositis,
Devic's
disease (neuromyelitis optica), discoid lupus, Dressler's syndrome,
endometriosis,
eosinophilic esophagitis (EoE), eosinophilic fasciitis, erythema nodosum,
essential mixed
cryoglobulinemia, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant
cell arteritis
(temporal arteritis), giant cell myocarditis, glomerulonephritis,
proliferative
glomerulonephritis, membranous nephropathy, minimal change nephrotic syndrome,
Goodpasture's syndrome, granulomatosis with polyangiitis, Graves' disease,
Guillain-Barre
syndrome, Hashimoto's thyroiditis, hemolytic anemia, immune hemolytic anemia,
Henoch-
Schonlein purpura (HSP), Herpes gestationis or pemphigoid gestationis (PG),
hypogammalglobulinemia, IgA Nephropathy, IgG4-related sclerosing disease,
Immune
thrombocytopenic purpura (ITP), Inclusion body myositis (IBM), Interstitial
cystitis (IC),
juvenile arthritis, juvenile diabetes (Type I diabetes), juvenile myositis
(JM), Kawasaki
disease, Lambert-Eaton syndrome, Leukocytoclastic vasculitis, Lichen planus,
Lichen
sclerosus, Ligneous conjunctivitis, Linear IgA disease (LAD), Lupus, Lyme
disease chronic,
Meniere's disease, Microscopic polyangiitis (MPA), Mixed connective tissue
disease
(MCTD), Mooren's ulcer, Mucha-Habermann disease, Multiple sclerosis,
Myasthenia gravis,
Myositis, Narcolepsy, Neuromyelitis optica, Neutropenia, Ocular cicatricial
pemphigoid,
Optic neuritis, Palindromic rheumatism (PR) PANDAS, Paraneoplastic cerebellar
degeneration (PCD), Paroxysmal nocturnal hemoglobinuria (PNH), Parry Romberg
syndrome, Pars planitis (peripheral uveitis), Parsonnage-Tumer syndrome,
Pemphigus,
peripheral neuropathy, perivenous encephalomyelitis, Pernicious anemia (PA),
POEMS
syndrome, Polyarteritis nodosa, Polyglandular syndromes type I, II, III,
Polymyalgia
rheumatica, Polymyositis, Postmyocardial infarction syndrome,
Postpericardiotomy
syndrome, Primary biliary cirrhosis, Primary sclerosing cholangitis,
Progesterone dermatitis,
Psoriasis, Psoriatic arthritis, Pure red cell aplasia (PRCA), Pyoderma
gangrenosum,
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Raynaud's phenomenon, Reactive arthritis, Reflex sympathetic dystrophy,
Relapsing
polychondritis, Restless legs syndrome (RLS), Retroperitoneal fibrosis,
Rheumatic fever,
Rheumatoid arthritis, Sarcoidosis, Schmidt syndrome, Scleritis, Scleroderma,
Sjogren's
syndrome, Sperm & testicular autoimmunity, Stiff person syndrome (SPS),
Subacute
bacterial endocarditis (SBE), Susac's syndrome, Sympathetic ophthalmia (SO),
Takayasu's
arteritis, Temporal arteritis/Giant cell arteritis, Thrombocytopenic purpura
(TTP), Tolosa-
Hunt syndrome (THS), Transverse myelitis, Type 1 diabetes, Ulcerative colitis
(UC),
Undifferentiated connective tissue disease (UCTD), Uveitis, Vasculitis,
Vitiligo, Wegener's
granulomatosis (or Granulomatosis with Polyangiitis (GPA)), idiopathic
thrombocytopenia
.. purpura, splenomegaly;
(xxii) Systemic inflammatory response syndrome, cytokine release syndrome,
cytokine storm, immune reaction to a drug(s) or therapy(s), immune reaction to
an immune
activating drug(s) or agent(s) or treatment(s) or intervention(s), immune
reaction to
immunotherapy and/or immune-oncology and/or immunomodulatory drug(s) and/or
treatment(s), adverse reaction to adoptive T-cell therapy(s), adverse reaction
to a chimeric
antigen receptor T-cell therapy(s) (CAR-T cell therapy(s)), adverse reaction
to a immune
checkpoint inhibitor(s), adverse reaction to monoclonal antibody drug(s),
tumor lysis
syndrome; or
(xxiii) cancer and Graft Versus Host Disease (GVHD) in transplantation therapy
in a
cancer patient;
wherein the method comprises administering to the subject an effective amount
of at
least one compound or composition from Claim 1.
1171 A method of treating, ameliorating, preventing, reversing or combating a
disease or
disorder, or unwanted/undesirable physiological process or its consequences or
an
unwanted/undesirable aesthetic, in a subject, selected from
(i) cancer that metabolizes much of its glucose and/or glutamine to lactate,
for
example a cancer exhibiting the Warburg effect and/or a cancer that can be
discriminated
from surrounding tissue by PET imaging (e.g. 18F-FDG PET);
(ii) cachexia, cancer driven cachexia, cancer fatigue, weight loss, weight
loss for
known or unknown reason, chronic wasting disease, atrophy, brown atrophy,
frailty, frailty
syndrome, aging frailty, geriatric syndrome, age-related cachexia and/or
sarcopenia,
weakness, wasting, anorexia nervosa, bulimia nervosa, eating disorder,
amenorrhea,
underweight, low body mass index (BMI, e.g. <18.5), low body fat percentage,
body
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composition change, wasting syndrome, HIV wasting syndrome, malnutrition,
clinical
malnutrition, starvation, kwashiorkor syndrome, marasmus syndrome,
malabsorption,
malabsorption due to parasitic/bacterial infection (e.g. helminthiasis,
Whipple's disease, small
intestine bacterial overgrowth (SIB0), giardiasis etc.), anemia, refeeding
syndrome, appetite
loss, catabolysis, muscle atrophy, asthenia, muscle weakness (myasthenia),
sarcopenia,
osteoporosis, cachexia associated with HIV, AIDS, multiple sclerosis,
rheumatoid arthritis,
familial amyloid polyneuropathy, chronic kidney disease, cystic fibrosis,
multiple sclerosis,
motor neuron disease, Parkinson's disease, dementia, Addison's disease,
mercury poisoning
(acrodynia), chronic pancreatitis, untreated/severe type I diabetes mellitus,
hormonal
deficiency, tuberculosis, gastroenteritis, diarrhea, dysentery, any digestive
disease or
disorder, any gastrointestinal disease or disorder including functional
gastrointestinal
disorders, coeliac disease, tropical sprue, irritable bowel syndrome,
inflammatory bowel
disease, Crohn's disease, ulcerative colitis, short bowl syndrome, congestive
heart failure,
constrictive pericarditis, bradycardia, chronic obstructive pulmonary disease
(COPD),
altitude sickness, hyperthyroidism (subclinical hyperthyroidism, Graves'
disease,
multinodular goiter, toxic adenoma, inflammation of the thyroid {thyroiditis},
pituitary
adenoma), fatigue, chronic fatigue syndrome or any disease or disorder or
pathology in which
a body tissue(s) is undersupplied or underutilises (vs. its need) an
energetic/chemical
substrate(s), including 02;
(iii) cancer associated fever, which is especially associated with, but not
limited to,
non Hodgkin lymphoma (NHL), Hodgkin lymphoma, acute leukaemia, kidney cancer
(renal
cell cancer), liver cancer (hepatocellular carcinoma), bone cancer, adrenal
gland tumours
such as phaeochromocytomas, tumours in the hypothalamus, solid tumours;
(iv) disease or disorder or physiological process or condition that causes a
higher than
.. normal body temperature such as (without limitation) high environmental
temperature,
ingesting an uncoupler (e.g. 2,4-dinitrophenol), infection, sepsis,
neutropenic sepsis, stroke,
fever, pyrexia, hyperpyrexia, hyperthermia, malignant hyperthermia,
neuroleptic malignant
syndrome, serotonin syndrome, toxic serotonin syndrome, thyroid storm,
heatstroke, surgery
related, menopause ("hot flushes"), infection (non-limiting e.g. roseola,
measles, enteroviral
.. infections, parasitic, viral, fungal, Chlamydial, Rickettsial, bacterial,
mycobacterial, systemic
bacterial, intravascular, FIIV associated, nosocomial), pyrogenic infection,
thermoregulatory
disorder(s), connective tissue disease(s), Kawasaki syndrome, drug overdose,
drug or drug
withdrawal induced hyperthermia, alcohol/drug withdrawal, idiosyncratic drug
reaction, fever
of known or unknown or uncertain origin (non-limiting e.g. infectious
disease(s),
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inflammation, immunological disease(s), non-infectious inflammatory disease(s)
{non-
limiting eg. systemic rheumatic and autoimmune diseases, vasculitis,
granulomatous diseases,
autoinflammatory syndromes), tissue destruction, reaction to incompatible
blood product(s),
metabolic disorder(s), inherited metabolic disorder(s), cancer, neoplasm,
endogenous or
exogenous pyrogen(s), injury, head injury);
(v) disease/disorder/injury/pathology/surgery
treatable/ameliorated/prevented/combated/helped by conferring hypothermia in a
subject for
some medical or other purpose which can include slowing a chemical reaction(s)
rate in a
subject for therapeutic benefit, preventing/minimizing brain and/or tissue
damage, slowing
physiological/pathological processes (reaction rates are temperature
dependent) and so
"buying time" to get the subject to treatment for their emergency (e.g.
trauma/septic shock or
other medical emergency), slowing the progress of sepsis until a sufficient
concentration of a
working antibiotic(s) can be built up in the subject (furthermore hypothermia,
by slowing
sepsis progression, buys time to observe which antibiotic(s) can work,
yielding time to try
.. alternative further antibiotic option(s) if required), used soon after or
just before clinical/legal
death to preserve the subject's organs/tissues until the subject can be
frozen/cryogenically
frozen or the pathology that caused clinical/legal death (e.g. wound) can be
fixed and the
subject resuscitated, administered to a subject when a first responder (e.g.
ambulance crew)
deems the subject dead or unlikely to survive the journey to a medical
facility (e.g. hospital)
wherein this administration helps to preserve the subject which is helpful if
hospital staff
subsequently assess that they can, or might be able to, save the subject,
stabilizing
surgical/trauma/Emergency Room (ER) patients, deep hypothermic circulatory
arrest for
surgery (DHCA, non-limiting applications of DHCA include repairs of the aortic
arch, repairs
to head and neck great vessels, repair of large cerebral aneurysms, repair of
cerebral
arteriovenous malformations, pulmonary thromboendarterectomy, resection of
tumors that
have invaded the vena cava, brain tumor resection {wherein the anti-cancer
activity of a
compound(s) of this invention juxtaposes well)), Emergency Preservation and
Resuscitation
(EPR), hypothermia for a surgical purpose, protective hypothermia during
surgery and/or
surgery complication, hypothermia to slow/reduce blood loss, hypothermia for
neuro- and/or
cardio- and/or organ/tissue and/or life protection in a subject that has
trauma/brain
trauma/polytrauma/surgery/stroke/ischemic stroke/hemorrhagic stroke/cardiac
arrest/myocardial infarction/hypoxia/shock (including, without limitation, low
volume,
cardiogenic, obstructive, and distributive shock)/sepsis/septic shock/multiple
organ
dysfunction syndrome/systemic inflammatory response syndrome (SIRS)/organ
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failure/cytokine storm/anaphylactic shock/seizure/disseminated intravascular
coagulation/blocked
airway/croup/rhabdomyolysis/[head/facial/spinal/chest/abdominal/ballistic/knife
injury/trauma], or some other medical
emergency/condition/disorder/disease/injury/operation,
.. hypothermia for cardiac and/or cardiovascular surgery and/or open heart
surgery and/or brain
surgery (neurosurgery) and/or surgery using total circulatory arrest and/or
surgery using
cardiopulmonary bypass, Emergency Preservation and Resuscitation (EPR),
preserving
detached body parts such as limbs and/or organs (for example during organ
storage/transport
and/or transplant, thus increasing the time window for transplantation of
organs to recipients;
compound(s) of this invention is administered to organ to be transplanted [by
administration
to donor and/or by administration to isolated organ] and/or to organ
recipient, optionally
during transplant operation), protective hypothermia, targeted temperature
management,
therapeutic hypothermia, hypothermia therapy for neonatal encephalopathy,
neonatal
hypoxia-ischemia, birth asphyxia, hypoxic-ischemic encephalopathy (I-11E),
haemorrhage,
hypovolemia, exsanguination, suspended animation, decompression sickness, burn
injury(s)
including skin burn, inflammation, allergic reaction, anaphylaxis,
tissue/organ rejection,
hypoxia, hypoxemia, anoxemia, anoxia, anemia, hypervolemia, altitude sickness,
obstructed
airway, asthma attack, hypoxia in a body/tissue/organ, hypoglycemia,
reperfusion injury
(ischemia-reperfusion injury), upon release of a ligature or tourniquet,
uraemia, crush
syndrome, compartment syndrome, traumatic brain and/or spinal cord injury,
major trauma,
infection, bacterial and/or viral infection(s) (non-limiting e.g. meningitis),
sepsis, septic
shock, stroke, cerebrovascular disease, ischemic brain injury, ischemic
stroke, traumatic
injury, brain injury, spinal cord injury, cardiac arrest, heart failure,
congestive heart failure,
Dilated cardiomyopathy, valvular heart disease, pulmonary embolism, adrenal
crisis,
Addisonian crisis, hypertensive emergency, haemorrhagic (hypovolemic) shock,
cardiogenic
shock, neurogenic shock, hepatic encephalopathy, blood loss, ischemic
brain/heart/kidney/intestinal injury, neuroprotection and/or cardioprotection
and/or tissue
protection during/after a stroke and/or ischemia and/or cardiac arrest and/or
resuscitation
and/or a period(s) of poor blood flow anywhere in a subject;
(vi) poisoning by a toxic amount of a compound(s) in a subject (non-limiting
e.g.
carbon monoxide/methanol/heavy metal/ethylene glycol/pesticide poisoning,
snake/spider/bee/insect/lizard venom, metabolic poison(s), nerve agent,
chemical weapon,
bacterial toxin(s) (e.g. food poisoning, Salmonella poisoning), endotoxemia,
eukaryote
produced toxin(s) e.g. (non-limiting) brevetoxin, drug(s)/substance(s)
overdose e.g. (non-
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limiting) heroin, ethanol, a prescription medication(s), an over the counter
medication(s) such
as aspirin, paracetamol etc.; hypothermia is protective to toxic insult);
(vii) hypermetabolism (optionally because of one or more of, without
restriction,
traumatic brain injury, injury to the body, infection, sepsis, burn, multiple
trauma, fever,
long-bone fracture, hyperthyroidism, prolonged steroid therapy, surgery, bone
marrow
transplant, recovery from anorexia/bulimia), heat intolerence, insomnia, fatal
insomnia,
nervousness, Luft's disease, non-thyroidal hypermetabolism, thyrotoxicosis,
hyperthyroidism,
overactive thyroid, subclinical hyperthyroidism, too much thyroid hormone(s)
in the subject,
too much triiodothyronine (T3) and/or thyroxine (T4) in the subject,
hyperthyroxinemia
(including, without restriction, familial dysalbuminemic hyperthyroxinemia,
familial
euthyroid hyperthyroxinemia, thyroid hormone resistance syndrome), thyroid
storm,
hyperthyroidism caused by one or more of (without restriction) Graves'
disease, thyroiditis,
Hashimoto's thyroiditis, subacute thyroiditis, postpartum thyroiditis, lumps
(nodules) on the
thyroid, enlarged thyroid gland (goitre), simple goitre, multinodular goiter,
toxic multinodular
goiter, toxic adenoma, toxic thyroid adenoma, inflammation of the thyroid,
hyperplasia of
thyroid, metastatic thyroid cancer, thyroid tumour, thyroid cancer (including,
without
restriction, papillary carcinoma, follicular carcinoma, medullary thyroid
carcinoma,
anaplastic thyroid carcinoma), eating too much iodine, goitrogen ingestion,
consumption of
ground beef contaminated with thyroid tissue ("hamburger hyperthyroidism"),
too much
synthetic thyroid hormone in the subject, pituitary adenoma, drug induced,
Amiodarone drug
induced, struma ovarii, Jod-Basedow syndrome, nonautoimmune autosomal dominant
hyperthyroidism;
(viii) low or less than desired metabolic/bioenergetic efficiency in a
subject, or low or
less than desired physical or mental performance (e.g. memory, IQ), or low or
less than
desired body weight, or fatigue/tiredness/weakness/exhaustion;
(ix) accelerated aging disease or progeroid syndrome including, without
restriction,
Werner syndrome, Bloom syndrome, De Barsy syndrome, Rothmund¨Thomson syndrome,
Cockayne syndrome, xeroderma pigmentosum, trichothiodystrophy, combined
xeroderma
pigmentosum-Cockayne syndrome, restrictive dermopathy, Wiedemann¨Rautenstrauch
syndrome, Hutchinson¨Gilford progeria syndrome (progeria), Ataxia
telangiectasia-like
disorder 2, XFE progeroid syndrome, Muscular dystrophy, Muscular Dystrophy
(Becker's,
Duchenne, Limb-Girdle), Yamamoto's Muscular Dystrophy, Mandibuloacral
dysplasia,
Dilated cardiomyopathy, GAPO syndrome, Cutis laxia, Ehlers-Danlos syndrom,
Lenz-
Majewski hyperostatic dwarfism, SHORT syndrome, Progressive external
opthalmoplegia,
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Nester-Guillermo progeria syndrome, MDPL syndrome, Dyskeratosis congenital,
Down
syndrome;
(x) disease or disorder of aging (incidence/severity increases with increased
age/senescence) and/or unwanted/undesirable aspect(s) of aging and/or a
disease/disorder
associated with elevated reactive oxygen species including age-associated
decline, aging
frailty, frailty syndrome, sarcopenia, muscle weakness, osteoporosis,
cognitive decline,
cognitive defecit, mild cognitive impairment, degenerative diseases,
neurodegenerative
diseases, motor-associated neurodegenerative diseases, motor neuron disease,
amyotrophic
lateral sclerosis (ALS), primary lateral sclerosis, progressive muscular
atrophy, progressive
bulbar palsy, progressive supranuclear palsy, pseudobulbar palsy, hereditary
spastic
paraplegia, Parkinson's disease, Multiple System Atrophy (MSA), Progressive
Supranuclear
Palsy (PSP), essential tremor, resting tremor, Alzheimer's disease,
Huntington's disease,
spinocerebellar ataxias, Friedreich's ataxia, dementia, frontotemporal
dementia, chronic
traumatic encephalopathy, memory loss, aged cognition, age/aging related
cognitive
decline/impairment, Batten disease, polyglutamine diseases, osteoporosis,
atherosclerosis,
cardiovascular disease, myocardial infarction, cerebrovascular disease,
stroke, heart failure,
heart failure with preserved ejection fraction, idiopathic pulmonary fibrosis,
fibrotic disease,
pulmonary disease, coronary artery disease, hypercholesterolemia, obesity,
liver disease, fatty
liver disease, lysosomal storage disease, amyloidosis, systemic sclerosis,
kidney disease,
hepatic cirrhosis, immunosenscence, clonal hematopoiesis, chronic obstructive
pulmonary
disease (COPD), hypertension, hypercholesterolemia, age-related thymic
atrophy, arthritis,
osteoarthritis, arthritis (Osteo-and Rheumatoid), Juvenile Rheumatoid
Arthritis (JRA),
Degenerative Disc Disease, Tendinopathy, Androgenetic Alopecia, male-pattern
baldness,
Idiopathic Pulmonary Fibrosis, systemic sclerosis, Chronic Obstructive
Pulmonary Disease,
Psoriasis, age-related loss of cardiac/pulmonary/cognitive/vision function,
diabetes, type 2
diabetes, andropause, glaucoma, retinal degeneration, sarcopenia, cachexia,
age-related
cachexia and/or sarcopenia, age-related macular degeneration (AMD,
early/intermediate/late), neovascular/wet AMD, dry AMD, Geographic atrophy
(GA), wet
and dry AMD in the same eye(s), Stargardt's macular degeneration, Best
vitelliform macular
dystrophy, diabetic retinopathy, proliferative diabetic retinopathy, diabetic
macular edema,
age/aging-related eye disease, ophthalmological disease/disorder, ocular
disease, vision loss,
progressive vision impairment, myopia (short-sightedness), degenerative
myopia, hyperopia
(far-sightedness), accommodative dysfunction, glaucoma, cataract formation,
retinal
degeneration, progressive retinal degeneration, retinitis pigmentosa, leber
hereditary optic
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CA 3050553 2019-07-25
neuropathy, Fuchs spot, Best's disease, Sorsby's fundus dystrophy, hearing
loss (e.g. age-
related), presbycusis, tinnitus, naive T cell shortage, movement disability,
nonalcoholic fatty
liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), immunosenescence,
respiratory/urinary tract infection (Rh/UT!) especially in older/aged/elderly
subjects, cancer;
(xi) aging and/or one or more signs of aging, wherein one or more of these
compounds slow/delay/reduce/treat/prevent/stop/reverse aging, and/or extend
lifespan and/or
healthspan, and/or treat or delay the onset of geriatric aging of the
human/animal body,
tissue(s), or organ(s), and/or treat or delay the onset of an age-associated
phenotype in a
cell(s)/organism(s), and/or prolong fertility e.g. female fertility, delay
menopause;
(xii) skin aging and/or damage (including sun damage) and/or scalp and/or hair
aging
and/or hair greying and/or hair loss;
(xiii) insomnia, fatal insomnia, sleep onset latency, delayed sleep phase
disorder,
exploding head syndrome, parasomnia, sleep-maintenance insomnia, sleep
disorder, too
much/inappropriate/undesired signals/activity/electrical activity in the
nervous system,
hyperactivity, hypersensitivity, Premature ejaculation, hyperreflexia,
Autonomic dysreflexia
(AD), Hyperventilation syndrome, brain hyperactivity, overly sensitive sensory
system,
pathological crying and/or laughing, Pseudobulbar affect (PBA, emotional
lability),
photophobia, phonophobia, temperature-sensitive, pressure-sensitive, brain
hyperexcitability,
overstimulation, intrusive thought(s), Perseveration, sensory overload,
disorganized thinking,
.. fantasy prone personality, malapdative daydreaming, dissociation,
hyperkinetic disorder,
agitation, Psychomotor agitation, restlessness, difficulty controlling
behaviour, disruptive
behaviour disorder, Emotional and behavioral disorder, pervasive developmental
disorder,
Overactive disorder associated with mental retardation and stereotyped
movements, attention-
deficit disorder, Attention Deficit Hyperactivity Disorder (ADHD), adult
attention-deficit
.. hyperactivity disorder, severe behavioral problem(s) in children (e.g., to
illustrate and not
restrict, combativeness and/or explosive hyperexcitable behavior {out of
proportion to
immediate provocation[s]}, hyperactive children who show excessive motor
activity with
accompanying conduct disorders consisting of one or more of: impulsivity,
difficulty
sustaining attention, aggressivity, mood lability, poor frustration
tolerance), Premenstrual
dysphoric disorder (PMDD), premenstrual syndrome (PMS), impulsiveness,
impulsivity,
impulse control disorder, lack of self-control, hysteria, histrionic
personality disorder,
attention difficulty, inattention, poor attention control, anxiety, paranoid
anxiety, Paranoid
personality disorder, distress, dysphoria, Adjustment disorder, separation
anxiety, anxiety
disorder, depressive anxiety, agitated depression, treatment-resistant
depression, Generalized
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CA 3050553 2019-07-25
anxiety disorder, social anxiety disorder, stranger anxiety, separation
anxiety (e.g. in dogs left
at home), separation anxiety disorder, Mixed anxiety-depressive disorder,
depression (all
forms, all severities), restlessness/apprehension/anxiety before surgery.
hypochondria, panic
disorder, panic attack, emotional outburst, emotional instability,
Intermittent explosive
disorder, unreasonable/unwarranted anger/aggression, hyper-aggression,
hostility, rage, poor
temper control, self-hatred, poor attentional control, worry, irritability,
neuroses, somatization
disorder, somatic symptom disorder, pain disorder, psychological pain,
psychogenic pain,
psychogenic facial pain, Atypical odontalgia (AO), burning mouth syndrome,
throbbing,
toothache/pulpitis/dental pain, chronic lower back pain, negative emotion,
persistent/enduring
negative emotion, body dysmorphic disorder, factitious disorder, illness
anxiety disorder,
unwarrented fight-or-flight response, stress, emotional stress, emotional
dysregulation,
distress, psychological stress, acute stress, chronic stress, acute stress
reaction, combat stress
reaction, traumatic grief, grief, grief after death of loved one, Prolonged
grief disorder (PGD),
heartbreak, guilt, shame, remorse, emotional pain, Algopsychalia, psychalgia,
suffering,
emotional trauma, psychological trauma, broken heart, Post Traumatic Stress
Disorder
(PTSD), Complex post-traumatic stress disorder (C-PTSD), hypervigilance,
sympathetic
hyperactivity, inability or impaired ability to relax, flashbacks, dysphoric
hyperarousal,
agoraphobia, insomnia, fatal insomnia, mood disorder, irrational/unwarrented
fear/terror,
phobia, social phobia, Cancerophobia, thunderstorm/firework phobia,
hypersexuality,
hypersexual disorder, depression, clinical depression, unipolar depression,
bipolar disorder,
Bipolar I, Bipolar II, Bipolar disorder not otherwise specified (Bipolar NOS),
cyclothymia,
cyclothymic disorder, mixed affective state, atypical depression, melancholic
depression,
postpartum depression, double depression, seasonal affective disorder, mania,
manic episode,
hypomania, increase in energy of psychomotor activity, delirium, excited
delirium, major
depressive disorder, minor depressive disorder, recurrent brief depression,
Depressive
Disorder Not Otherwise Specified (DD-NOS), major depressive episode,
persistent
depressive disorder (PDD), dysthymia, dysthymic disorder, absence of euthymia,
manic
thoughts, racing thoughts, thought disorder, disordered thinking, reduced
ability to plan and
execute tasks, paranoia, hallucination (including, without limitation, visual,
auditory,
olfactory, gustatory, tactile, proprioceptive, equilibrioceptive, nociceptive,
thermoceptive,
chronoceptive), Charles Bonnet syndrome, delusion, persecutory delusion,
hearing voices,
homicidal/criminal ideation/tendency/thoughts, suicidal
ideation/tendancy/thoughts, self-
injury, non-suicidal self-injury, violence, attacking others, negative mood
swing, personality
disorder, Borderline personality disorder, Narcissistic personality disorder,
malignant
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CA 3050553 2019-07-25
narcissism, dissociative disorder, dissociative identity disorder (DID),
Psychosis, acute
psychosis, chronic psychosis, psychosis spectrum disorder, manifestations of
psychotic
disorders, behavioral complications of mental retardation, stimulant
psychosis, psychotic
depression, hallucinogen persisting perception disorder, Psychoactive
substance-related
disorder, amphetamine-induced psychosis, brief psychotic disorder, Brief
reactive psychosis,
Menstrual psychosis, postpartum psychosis, Psychotic disorder, Psychopathy,
chronic
hallucinatory psychosis, manifestation(s) of psychotic disorder,
neurotic/reactive/endogenous/involutional/psychotic depression/depressive
disorder
(optionally accompanied by anxiety or agitation), depressive neurosis,
delusional depression,
psychotic aggression, psychiatric symptoms of dementia, AIDS delirium,
Supersensitivity
psychosis, Tardive psychosis, Tardive dysmentia, Depersonalization disorder,
out-of-body
experience, Sociopathy, Schizophrenia, Paranoid schizophrenia, disorganized-
type
schizophrenia, simple-type schizophrenia, pseudoneurotic schizophrenia,
prodromal
schizophrenia, schizoaffective disorder, bipolar type schizoaffective
disorder, depressive type
schizoaffective disorder, schizoaffective psychosis, Schizotypal personality
disorder,
schizophreniform disorder, Delusional parasitosis, formication, paresthesias,
Acroparesthesia,
tinnitus, delusional disorder, delusional jealousy, inappropriate behaviour,
behavioural
disorder, antisocial personality disorder, Oppositional defiant disorder
(ODD), conduct
disorder (CD), Disruptive mood dysregulation disorder (DMDD), sadistic
personality
disorder, poor inhibitory control, kleptomania, Pyromania, trichotillomania,
dermatillomania,
pathological/problem gambling, dyskinesia, tardive dyskinesia, paroxysmal
dyskinesia,
Paroxysmal kinesigenic dyskinesia, Paroxysmal nonkinesigenic dyskinesia,
Paroxysmal
exercise-induced dystonia, Hemiballismus, tic disorder, tremor, clonus,
Tourette's syndrome,
coprolalia, copropraxia, Echophenomena, Echopraxia, Echolalia, Palilalia,
stuttering/stammering, stereotypy, punding, Self-stimulatory behaviour
(stimming),
Stereotypic movement disorder (SMD), synesthesia, obsession,
Obsessive¨compulsive
disorder (OCD), obsessive¨compulsive personality disorder, anankastic
personality disorder,
relationship obsessive¨compulsive disorder (ROCD), Scrupulosity, Primarily
obsessional
obsessive compulsive disorder, sexual obsession, Akathisia (including, without
limitation,
chronic, acute, Pseudoakathisia, Tardive akathisia, withdrawal or "rebound"
akathisia),
Restless legs syndrome, motor restlessness, periodic limb movement disorder
(PLMD),
periodic limb movements in sleep (PLMS), Sydenham's chorea, chorea, dystonia,
Hypnic
jerk, twitching, spasms, Tetanus, tetanus muscle spasms, tetanized state,
Myoclonus,
myoclonic seizure, Action myoclonus, Palatal myoclonus, Middle ear myoclonus,
Spinal
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CA 3050553 2019-07-25
myoclonus, Stimulus-sensitive myoclonus, Sleep myoclonus, Cortical reflex
myoclonus,
Essential myoclonus, myoclonic epilepsy, Juvenile myoclonic epilepsy,
Progressive
myoclonus epilepsy (PME, including, without limitation, Dentatorubral-
pallidoluysian
atrophy, Unverricht-Lundborg disease, MERRF syndrome, Lafora disease),
Reticular reflex
myoclonus, Myoclonic epilepsy, diaphragmatic flutter, automatism, status
epilepticus,
Epilepsia partialis continua, Complex partial status epilepticus, epilepsy,
epileptic seizure,
simple partial seizure, complex partial seizure, generalized epilepsy,
generalized seizure
(including, without limitation, tonic-clonic, tonic, clonic, myoclonic,
absence (including
typical absence and atypical absence), atonic seizure), focal epilepsy, focal
seizure,
focal/partial seizure (including, without limitation, Simple partial seizure
and Complex partial
seizure), focal aware seizure, focal impaired awareness seizure, generalised
epilepsy,
temporal lobe epilepsy (including, without restriction, mesial temporal lobe
epilepsy
{MTLE} and lateral temporal lobe epilepsy {LTLE}), Frontal lobe epilepsy,
Rolandic
epilepsy, Nocturnal epilepsy, Nocturnal frontal lobe epilepsy, Autosomal
dominant nocturnal
frontal lobe epilepsy (ADNFLE), Generalized epilepsy with febrile seizures
plus (GEFS+),
Panayiotopoulos syndrome, epileptic fit, reflex epilepsy, reflex seizure,
absence seizure
(including, without limitation, childhood absence epilepsy, epilepsy with
myoclonic
absences, juvenile absence epilepsy, juvenile myoclonic epilepsy, Jeavons
syndrome {eyelid
myoclonia with absences}, genetic generalised epilepsy with phantom absences),
complex
partial seizure, atonic seizure, generalized tonic-clonic seizure,
tonic¨clonic seizure, extrinsic
stimulus epilepsy, intrinsic stimulus epilepsy, photosensitive epilepsy,
musicogenic epilepsy,
thinking epilepsy, eating epilepsy, seizure(s), Febrile seizure, nerve agent
induced seizure,
Dravet syndrome (sometimes modest hyperthermic stressors like physical
exertion or a hot
bath can provoke seizures in affected individuals), acute symptomatic seizure,
seizure-related
.. disorder, drug related seizure, paroxysmal depolarizing shift, Ohtahara
syndrome, Epilepsy in
females with mental retardation, Rasmussen's encephalitis, Epilepsy syndrome,
benign
rolandic epilepsy, childhood absence epilepsy, absence epilepsy, juvenile
myoclonic
epilepsy, epileptic encephalopathies, Lennox¨Gastaut syndrome, West syndrome
(Epileptic
spasms), Doose syndrome (Myoclonic astatic epilepsy, MAE), Lennox-Gestaut
syndrome,
pseudo-Lennox Gastaut syndrome (atypical benign partial epilepsy), Benign
familial neonatal
epilepsy, Benign occipital epilepsy of childhood, familial neonatal
convulsions, Febrile
infection-related epilepsy syndrome, interictal dysphoric disorder, euphoria
sclerotic,
psychogenic non-epileptic seizure, non-epileptic seizure, gelastic seizure,
convulsion(s),
migraine, status migrainosus, tension headache, headache, Hypnic headache,
hiccups,
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CA 3050553 2019-07-25
intractable hiccups, thumps in equines, Postural orthostatic tachycardia
syndrome (POTS),
Pheochromocytoma, agony, pain, chronic pain, acute pain, pain due to
disease/injury,
neuropathic pain, fibromyalgia, postherpetic neuralgia, phantom pain, referred
pain, back
pain, lower back pain, pelvic pain, cancer associated pain, chemotherapy
associated pain,
Diabetic neuropathy, small fiber peripheral neuropathy, Mononeuritis
multiplex,
Wartenberg's migratory sensory neuropathy, Breakthrough pain, idiopathic pain,
polyneuropathy, Neuritis, Mononeuropathy, Polyradiculoneuropathy, Radial
neuropathy,
neuropathy, visceral pain, Burning feet syndrome, Tarsal tunnel syndrome,
Carpal tunnel
syndrome, Guyon's canal syndrome, Cubital Tunnel Syndrome, Repetitive strain
injury,
Sciatica, Neurofibromatosis, Ulnar nerve entrapment, Erythromelalgia,
Paroxysmal extreme
pain disorder, Proctalgia fugax, dysesthesia, scalp dysesthesia, dysesthetic
burning,
hyperesthesia, hyperalgesia, Opioid-induced hyperalgesia, hyperpathia,
allodynia, pain
response from stimuli which do not normally provoke pain, Complex regional
pain syndrome
(said to be most painful condition known to man), Radiculopathy, neuralgia
(including,
without restriction, intercostal neuralgia, trigeminal neuralgia, atypical
trigeminal neuralgia,
glossopharyngeal neuralgia, occipital neuralgia, postherpetic neuralgia),
ciguatera poisoning,
irritable bowel syndrome (IBS), interstitial cystitis (IC), temporomandibular
joint disorder,
acute intermittent porphyria, Porphyria, Acute porphyria (including, without
limitation, acute
intermittent porphyria {AIP}, variegate porphyria {VP}, aminolevulinic acid
dehydratase
deficiency porphyria {ALAD}, hereditary coproporphyria {HCP}, drug induced),
Chronic
porphyria (including, without limitation, X-linked dominant protoporphyria
{XLDPP},
congenital erythropoietic porphyria {CEP}, porphyria cutanea tarda {PCT}, and
erythropoietic protoporphyria {EPP}), cutaneous porphyria, Porphyria cutanea
tarda, allergy,
allergic reaction, anaphylaxis, anaphylactic shock, hives, asthma, allergic
rhinitis, rhinitis,
urticaria, contact dermatitis, cough, sore throat, esophageal reflux disease,
heartburn, chest
pain, Esophageal motility disorder, Nutcracker esophagus, diseases involving
gastrointestinal
motility, Peptic ulcer disease, esophageal spasm, angina, itchiness, Pruritus,
severe pruritus,
Prurigo, Pruritic skin condition, chronic itch, eczema, pruritus in eczema,
neuropathic itch,
neurogenic itch, itchiness due to atopic dermatitis and/or lichen simplex
chronicus, peripheral
sensitization, central sensitization, sensory perception of absent stimuli,
too much sensory
stimulation, sensory stimulation brings discomfort, Neuromyotonia, Peripheral
nerve
hyperexcitability, Morvan's syndrome, Benign fasciculation syndrome, Cramp
fasciculation
syndrome, hyperhidrosis, undifferentiated somatoform disorder, somatoform
disorder,
somatic symptom disorder, conversion disorder, functional neurological symptom
disorder,
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CA 3050553 2019-07-25
severe nausea and/or vomiting, severe nausea/emesis, Chemotherapy-induced
peripheral
neuropathy, chemotherapy-induced nausea and vomiting (CINV), radiation therapy-
induced
nausea and vomiting (RINV), postnarcotic nausea, hyperemesis gravidarum,
morning
sickness, Cyclic vomiting syndrome, retching/dry heaving, Urinary
incontinence, enuresis,
nocturnal enuresis, tail chasing, reduce urine spraying/marking behavior,
benzodiazepine
withdrawal syndrome, barbituate withdrawal syndrome, Neuroleptic
discontinuation
syndrome, drug withdrawal discomfort/pain/symptoms, drug use disorder, alcohol
use
disorder, opoid use disorder, amphetamine use disorder, cocaine use disorder,
alcohol
withdrawal syndrome/symptoms, delirium tremens, opoid withdrawal
sydrome/symptoms,
drug craving, drug addiction, drug dependence, polysubstance dependence, drug
overdose,
smoking, tobacco (nicotine) addition, tobacco (nicotine) withdrawal symptoms,
alcoholism,
addiction, opoid addiction, cocaine/crack addiction, addictive behaviour,
addictive
personality, behavioural addiction, intemet/computer/computer game/social
media/media
addiction, exercise addiction, compulsive behaviour (e.g. {non-limiting}
checking, counting,
washing, repeating), anti-social behaviour, criminality, sexual compulsion,
impulsive sexual
behaviour, compulsive buying, gambling addiction, sex related addiction,
sexual urge,
hunger, eating desire/compulsion, eating disorder, polyphagia, overeating,
binge eating
disorder, compulsive overeating, insatiable/excessive appetite, bulimia
nervosa, anorexia
nervosa, substance abuse, substance-induced delirium, substance-induced
psychosis,
substance-induced mood disorder, drug overdose, vertigo, motion sickness,
seasickness,
mental/nervous breakdown, Autism spectrum disorder, neurological disorder,
cognitive
disorder, mental disorder, mental health disorder, mental health condition
involving impaired
or altered neural plasticity, mood disorder, mental disorder disclosed in
Diagnostic and
Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) or a later
edition, a
mental/behavioural disorder disclosed by the International Classification of
Diseases (ICD) in
ICD-10 Chapter V: Mental and behavioural disorders (World Health Organisation,
WHO); or
(xiv) diseases or disorders or conditions or pathologies or
unwanted/undesirable
effects/actions/behaviour treatable/ameliorated/prevented/combated, in
totality or in part (e.g.
along with surgery), by anaesthesia, pre-anaesthesia, post-anaesthesia,
hypoesthesia,
hypoactivity, sedation, coma, tranquilization, behavioural submission, muscle
relaxation,
hibernation, artificial hibernation, torpor, synthetic torpor, suspended
animation (e.g. used
during spaceflight because e.g. reduces ionizing radiation damage to subject);
(xv) hyperproliferative/hyperplasia disorder, non-cancerous proliferative
disorder,
hyperproliferative autoimmune disorder, hyperplasia, epidermal hyperplasia,
dysplasia (e.g.
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CA 3050553 2019-07-25
epithelial dysplasia), nodule(s), wart(s), papilloma(s), squamous cell
papilloma, genital
wart(s), condyloma(s), condyloma acuminatum, cyst(s), polyp(s) {including,
without
restriction, digestive, colorectal, endometrial, cervical, nasal, laryngeal,
inflammatory fibroid
polyp[s])}, inherited/hereditary (including, without restriction, Familial
adenomatous
polyposis, Peutz¨Jeghers syndrome, Turcot syndrome, Juvenile polyposis
syndrome, Cowden
disease, Bannayan¨Riley¨Ruvalcaba syndrome {Bannayan-Zonana syndrome},
Gardner's
syndrome) and non-inherited (non-restrictive e.g. Cronkhite¨Canada syndrome)
polyposis
syndrome, benign tumour, adenoma, organ enlargement by hyperplasia, Cushing's
disease
(enlarged adrenal cortex by hyperplasia), congenital adrenal hyperplasia,
hyperplasia of
breast, atypical ductal hyperplasia, intraductal papillomatosis,
fibroadenomas, fibrocystic
changes, hemihyperplasia, focal epithelial hyperplasia, sebaceous hyperplasia,
sebaceous
adenoma, intimal hyperplasia, unwanted/undesirable smooth muscle cell
proliferation,
smooth muscle cell hyperplasia, intimal smooth muscle cell hyperplasia,
neointimal
hyperplasia, proliferative vascular disorders, stenosis, stenosis because of
cellular
proliferation, vaginal stenosis, stenosis in a blood vessel, lessoned patency
of a blood vessel,
stenosis in a blood vessel because of cellular proliferation, vascular
occlusion, restenosis,
restenosis in a blood vessel that has been implanted with a stent, in-stent
restenosis, post-
angioplasty restenosis, systemic sclerosis, cirrhosis of the liver, adult
respiratory distress
syndrome, idiopathic cardiomyopathy, lupus erythematosus, retinopathy (e.g.
diabetic
retinopathy and/or other retinopathy[y/ies]), cardiac hyperplasia, fibrosis,
pulmonary fibrosis,
idiopathic pulmonary fibrosis, fibromatosis, neurofibromatosis, renal
interstitial fibrosis,
Cowden syndrome, hamartoma(s), choristoma(s), hemangioma(s), lymphangioma(s),
rhabdomyoma(s), lymphangiomatosis, cystic hygroma, trichilemmoma, sarcoidosis,
neurosarcoidosis, aggressive fibromatosis, desmoid tumour(s),
unwanted/undesirable skin
cell proliferation, hyperproliferative skin disorder, psoriasis (including,
without restriction,
plaque, guttate, inverse, pustular, napkin, seborrheic-like, nail, scalp and
erythrodermic
psoriasis), psoriatic arthritis, dactylitis, seborrhoeic dermatitis, dandruff,
eczema, atopic
dermatitis, rosacea, reactive arthritis (Reiter's syndrome), pityriasis rubra
pilaris,
hyperproliferative variants of disorders of keratinization (e.g., without
restriction, actinic
keratosis, senile keratosis, keratosis pilaris, seborrheic keratosis),
scleroderma, benign
prostatic hyperplasia, prostate enlargement, endometrial hyperplasia, atypical
endometrial
hyperplasia, benign endometrial hyperplasia, adenomyosis, atypical polypoid
adenomyoma,
endometriosis, endometriosis of ovary (endometrioma), endometrial polyp(s),
polycystic
ovary syndrome, ovarian cyst(s), cervical polyp(s), uterine fibroid(s),
uterine hyperplasia;
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CA 3050553 2019-07-25
(xvi) Tumour Associated Macrophages (TAMs) or any macrophage associated
disease or disorder such as, without limitation, Macrophage Activation
Syndrome (MAS),
HIV, AIDS, HIV-associated neurocognitive disorders (HAND), AIDS dementia, HIV
peripheral neuropathy, HIV associated cancers, AIDS-defining cancers, non-AIDS
defining
cancers, any disease in which the pathogen(s) hides from the immune system in
macrophages
including, without limitation, Mycobacterium tuberculosis (causes
tuberculosis), Leishmania
parasite (causes Leishmaniasis), Chikungunya virus (causes Chikungunya),
Legionella
pneumophila (causes Legionnaires' disease), adenoviruses, T. whipplei (causes
Whipple's
Disease), Brucella spp. (causes brucellosis), Staphylococcus aureus, Ebola
virus, Hepatitis B
virus, Hepatitis C virus, influenza virus strains, dengue virus and antibiotic
resistant bacteria,
any disease or condition in which activated macrophages are unwanted or
undesirable;
(xvii) virus/pathogen neuroinvasion via macrophage(s), as used for non-
limiting
example by HIV, Heptatitis C virus and SARS coronavirus;
(xviii) neurocognitive or neurodegenerative diseases/disorders, for non-
limiting
example those caused by a virus;
(xix) virus/pathogen transmission from mother to fetus/baby via macrophage(s)
as
used for non-limiting example by zika (via Hofbauer cells) and HIV
(macrophages in breast
milk);
(xx) acute or chronic or systemic inflammation or any inflammatory
disease/disorder/syndrome or any autoinflammatory disease/disorder/syndrome or
any
autoimmune disease/disorder/syndrome;
(xxi) acute inflammation, chronic inflammation, systemic inflammation,
inflammation
because of infection or foreign bodies or injury or chemical or toxin or drug
or stress or
frostbite or burn or ionising radiation or surgery, inflammatory
diseases/disorders/syndromes,
Macrophage Activation Syndrome (MAS), autoinflammatory
diseases/disorders/syndromes,
age-related chronic inflammatory diseases ("inflammaging"), autoimmune
diseases/disorders/syndromes, diseases/disorders of the innate immune system,
sore throat,
sore throat associated with cold or flu or fever, high-intensity exercise
associated
inflammation, ulcerative colitis, inflammatory bowel disease (IBD), irritable
bowel syndrome
(IBS), rheumatoid arthritis, osteoarthritis, inflammatory osteoarthritis,
psoriatic arthritis,
atopic dermatitis, allergic airway inflammation, asthma, inflammation
associated depression,
neuroinflammation, neuropathic pain, exercise-induced acute inflammation,
atherosclerosis,
allergy, hay fever, anaphylaxis, inflammatory myopathies, drug-induced
inflammation,
systemic inflammatory response syndrome, sepsis-related multiple organ
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CA 3050553 2019-07-25
dysfunction/multiple organ failure, microbial infection, acute
brain/lung/hepatic/renal
injuries, lung inflammation, acute lung injury (ARDS), acne vulgaris, celiac
disease, celiac
sprue, chronic prostatitis, colitis, autoimmune hemolytic anemia,
diverticulitis,
glomerulonephritis, proliferative glomerulonephritis, membranous nephropathy,
minimal
change nephrotic syndrome, hidradenitis suppurativa, hypersensitivities,
interstitial cystitis,
Mast Cell Activation Syndrome, mastocytosis, otitis, pelvic inflammatory
disease (PID),
endometritis, reperfusion injury, rheumatic fever, rhinitis, sarcoidosis,
transplant rejection,
parasitosis, eosinophilia, type III hypersensitivity, ischaemia, chronic
peptic ulcer,
tuberculosis, Crohn's disease, hepatitis, chronic active hepatitis, immune
hepatitis, alcoholic
hepatitis, chronic viral hepatitis, ankylosing spondylitis, diverticulitis,
fibromyalgia, systemic
lupus erythematous (SLE), Alzheimer's disease, Parkinson's disease,
neurodegenerative
disease, cardiovascular disease, chronic obstructive pulmonary disease,
bronchitis, acute
bronchitis, bronchiectasis, bronchopneumonia, obliterative bronchiolitis,
appendicitis, acute
appendicitis, bursitis, cystitis, dermatitis, encephalitis, HIV encephalitis,
gingivitis,
meningitis, infective meningitis, myelitis, nephritis, neuritis,
periodontitis, chronic
periodontitis, phlebitis, prostatitis, RSD/CRPS, rhinitis, sinusitis, chronic
sinusitis, tendonitis,
testiculitis, tonsillitis, urethritis, vasculitis, respiratory
bronchiolitis¨associated interstitial
lung disease and desquamative interstitial pneumonia, pneumonia, interstitial
lung disease,
Liifgren syndrome, Heerfordt syndrome, monocytosis, liver fibrosis,
steatohepatitis,
nonalcoholic steatohepatitis, silicosis, histiocytoses, Langerhans' cell
histiocytosis,
haemophagocytic lymphohistiocytosis, pulmonary langerhans cell histiocytosis,
obesity, type
II diabetes, gout, pseudogout, organ transplant rejection, epidermal
hyperplasia, chronic
fatigue syndrome, graft versus host disease (GVHD), lymphadenopathy,
rheumatoid arthritis
(RA), osteoarthritis (OA), inflammatory osteoarthritis, lupus, multiple
sclerosis (MS),
myocarditis, uveitis, CNS disease(s), inflammation aspect to a CNS disease(s),
hypothalamic
inflammation, dementia, glaucoma, amyloid related/driven disease, lipid
storage disease(s),
fibrosis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic
steatohepatitis (NASH),
cirrhosis, cirrhosis of the liver, alcoholic cirrhosis, nephropath(y/ies),
lupus nephritis,
immune nephritis, fibrotic disorder(s), cardiovascular disease, heart disease,
atherosclerosis,
vulnerable plaque, plaque formation, lipid containing macrophage related
disease(s)/disorder(s)/malad(y/ies), macrophage foam cells, diabetes, type 1
diabetes, type 2
diabetes, insulin resistance, macrophage aspect to insulin resistance,
obesity, obesity
associated inflammation, macrophage accumulation/large numbers of macrophages
in
adipose tissue (e.g. associated with obesity), granuloma(s), granulomatous
diseases,
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sarcoidosis (including, without limitation, Annular sarcoidosis, Erythrodermic
sarcoidosis,
Ichthyosiform sarcoidosis, Hypopigmented sarcoidosis, Lofgren syndrome, Lupus
pernio,
Morpheaform sarcoidosis, Mucosal sarcoidosis, Neurosarcoidosis, Papular
sarcoid, Scar
sarcoid, Subcutaneous sarcoidosis, Systemic sarcoidosis, Ulcerative
sarcoidos),
neurosarcoidosis, pulmonary sarcoidosis, interstitial lung disease, pulmonary
fibrosis,
pulmonary tuberculosis, immune reconstitution syndrome of HIV,
Jarisch¨Herxheimer
reaction, sepsis, Paget's disease of bone, osteolysis, monocytosis,
histiocytosis, X-type
histiocytoses, non-X histiocytoses, Langerhans cell histiocytosis, non-
Langerhans-cell
histiocytosis, malignant histiocytosis, malignant histiocytic disorders,
histiocytomas,
histiocytic lymphoma, hemophagocytic syndrome, hemophagocytic
lymphohistiocytosis,
lymphohistiocytosis, diffuse histiocytic sarcoma, Rosai¨Dorfman disease,
gliosis, Bergmann
gliosis, Chronic Obstructive Pulmonary Disease (COPD), chronic inflammatory
lung disease,
familial mediterranean fever (FMF), TNF receptor-associated periodic syndrome
(TRAPS),
Hyperimmunoglobulinemia D with recurrent fever syndrome (HIDS), cryopyrin
associated
periodic syndrome (CAPS), Blau syndrome, Majeed syndrome, deficiency of
interleukin-1
receptor antagonist (DIRA), mevalonate kinase deficiency, pyogenic-arthritis-
pyoderma
gangrenosum and acne syndrome (PAPA), periodic fever aphthous stomatitis
pharyngitis
adenitis (PFAPA) syndrome, Behcet's disease, Still's disease, Crohn's disease,
Schnitzler's
syndrome, Sweet's syndrome, NLRP12-associated autoinflammatory disorders,
deficiency of
interleukin-1 receptor antagonist (DIRA), pyoderma gangrenosum, cystic acne,
aseptic
arthritis, periodic Fever Associated with mevalonate kinase deficiency
(hyperimmunoglobulin D Syndrome), Pyogenic Arthritis Pyoderma Gangrenosum Acne
(PAPA) syndrome, Periodic Fever Aphthous Stomatitis, Pharyngitis and
Adenopathy
(PFAPA) syndrome, Adult-Onset Still's Disease (AOSD), Systemic Juvenile
Idiopathic
Arthritis (sJIA), Chronic Recurrent Multifocal Osteomyelitis (CRMO), Synovitis
Acne
Pustulosis Hyperostosis Osteitis (SAPHO) syndrome, Cryopyrin associated
Periodic
Syndrome (CAPS), Familial cold auto inflammatory syndrome (FCAS), Muckle-Wells
syndrome (MWS), Familial cold urticarial, Neonatal onset multisystemic
inflammatory
disorder (NOMID), hereditary Periodic Fever Syndromes, Periodic Fever
Syndromes,
systemic autoinflammatory diseases, Addison's disease, Agammaglobulinemia,
Alopecia
areata, Amyloidosis, Ankylosing spondylitis, Anti-GBM/Anti-TBM nephritis,
Antiphospholipid syndrome, autoimmune angioedema, autoimmune dysautonomia,
autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune inner ear
disease
(AIED), autoimmune myocarditis, autoimmune pancreatitis, autoimmune
retinopathy,
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CA 3050553 2019-07-25
autoimmune urticaria, axonal & neuronal neuropathy (AMAN), Balo disease,
Behcet's
disease, benign mucosal pemphigoid, Bullous pemphigoid, Castleman disease
(CD), Celiac
disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy
(CIDP),
chronic recurrent multifocal osteomyelitis (CRMO), Churg-Strauss, Cicatricial
pemphigoid,
.. Cogan's syndrome, cold agglutinin disease, congenital heart block,
coxsackie myocarditis,
CREST syndrome, Berger's disease, dermatitis herpetiformis, dermatomyositis,
Devic's
disease (neuromyelitis optica), discoid lupus, Dressler's syndrome,
endometriosis,
eosinophilic esophagitis (EoE), eosinophilic fasciitis, erythema nodosum,
essential mixed
cryoglobulinemia, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant
cell arteritis
(temporal arteritis), giant cell myocarditis, glomerulonephritis,
proliferative
glomerulonephritis, membranous nephropathy, minimal change nephrotic syndrome,
Goodpasture's syndrome, granulomatosis with polyangiitis, Graves' disease,
Guillain-Barre
syndrome, Hashimoto's thyroiditis, hemolytic anemia, immune hemolytic anemia,
Henoch-
Schonlein purpura (HSP), Herpes gestationis or pemphigoid gestationis (PG),
hypogammalglobulinemia, IgA Nephropathy, IgG4-related sclerosing disease,
Immune
thrombocytopenic purpura (ITP), Inclusion body myositis (IBM), Interstitial
cystitis (IC),
juvenile arthritis, juvenile diabetes (Type 1 diabetes), juvenile myositis
(JM), Kawasaki
disease, Lambert-Eaton syndrome, Leukocytoclastic vasculitis, Lichen planus,
Lichen
sclerosus, Ligneous conjunctivitis, Linear IgA disease (LAD), Lupus, Lyme
disease chronic,
Meniere's disease, Microscopic polyangiitis (MPA), Mixed connective tissue
disease
(MCTD), Mooren's ulcer, Mucha-Habermann disease, Multiple sclerosis,
Myasthenia gravis,
Myositis, Narcolepsy, Neuromyelitis optica, Neutropenia, Ocular cicatricial
pemphigoid, =
Optic neuritis, Palindromic rheumatism (PR) PANDAS, Paraneoplastic cerebellar
degeneration (PCD), Paroxysmal nocturnal hemoglobinuria (PNH), Parry Romberg
syndrome, Pars planitis (peripheral uveitis), Parsonnage-Tumer syndrome,
Pemphigus,
peripheral neuropathy, perivenous encephalomyelitis, Pernicious anemia (PA),
POEMS
syndrome, Polyarteritis nodosa, Polyglandular syndromes type I, II, III,
Polymyalgia
rheumatica, Polymyositis, Postmyocardial infarction syndrome,
Postpericardiotomy
syndrome, Primary biliary cirrhosis, Primary sclerosing cholangitis,
Progesterone dermatitis,
Psoriasis, Psoriatic arthritis, Pure red cell aplasia (PRCA), Pyoderma
gangrenosum,
Raynaud's phenomenon, Reactive arthritis, Reflex sympathetic dystrophy,
Relapsing
polychondritis, Restless legs syndrome (RLS), Retroperitoneal fibrosis,
Rheumatic fever,
Rheumatoid arthritis, Sarcoidosis, Schmidt syndrome, Scleritis, Scleroderma,
Sjogren's
syndrome, Sperm & testicular autoimmunity, Stiff person syndrome (SPS),
Subacute
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CA 3050553 2019-07-25
bacterial endocarditis (SBE), Susac's syndrome, Sympathetic ophthalmia (SO),
Takayasu's
arteritis, Temporal arteritis/Giant cell arteritis, Thrombocytopenic purpura
(TTP), Tolosa-
Hunt syndrome (THS), Transverse myelitis, Type I diabetes, Ulcerative colitis
(UC),
Undifferentiated connective tissue disease (UCTD), Uveitis, Vasculitis,
Vitiligo, Wegener's
granulomatosis (or Granulomatosis with Polyangiitis (GPA)), idiopathic
thrombocytopenia
purpura, splenomegaly;
(xxii) Systemic inflammatory response syndrome, cytokine release syndrome,
cytokine storm, immune reaction to a drug(s) or therapy(s), immune reaction to
an immune
activating drug(s) or agent(s) or treatment(s) or intervention(s), immune
reaction to
immunotherapy and/or immune-oncology and/or immunomodulatory drug(s) and/or
treatment(s), adverse reaction to adoptive 1-cell therapy(s), adverse reaction
to a chimeric
antigen receptor T-cell therapy(s) (CAR-T cell therapy(s)), adverse reaction
to a immune
checkpoint inhibitor(s), adverse reaction to monoclonal antibody drug(s),
tumor lysis
syndrome; or
(xxiii) cancer and Graft Versus Host Disease (GVHD) in transplantation therapy
in a
cancer patient;
wherein the method comprises administering to the subject an effective amount
of at
least one compound, or a composition containing at least one compound, of the
following
formula:
R2 R3
\N"
N/INN
L
R1
(4
(R4)
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
L is selected from alkyl, substituted alkyl, deuterated alkyl, aminoalkyl,
thioalkyl,
alkoxy, halogen, haloalkyl, haloalkoxy, hydroxyalkyl, or any atom or isotope
permitted by
valence (including any accompanying hydrogens by valence e.g. (non-limiting)
OH, NH2,
SH, SiH3, PH2 etc.);
RI is hydrogen, cyano, ¨SO2R8, ¨C(=0)R9, heteroaryl or thiazolyl;
R2 is (i) independently hydrogen, alkyl, benzyl, or substituted alkyl, or (ii)
taken
together with R3 forms a heterocyclo;
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CA 3050553 2019-07-25
R3 is (i) independently alkyl, substituted alkyl, alkylthio, aminoalkyl,
carbamyl, BB-
aryl, BB-heterocyclo, BB-heteroaryl, or BB-cycloalkyl, or (ii) phenyl
optionally substituted
with Ci_4alkyl, halogen, trifluoromethyl, OCF3, cyano, nitro, amino, hydroxy,
or methoxy, or
(iii) independently selected from C1_4alkyl, alkylthio, aminoalkyl, -BB-aryl, -
BB-heterocyclo,
BB-cycloalkyl, and -BB-hetaroaryl, optionally having one to three substituents
selected from
R3a; and/or having fused thereto a five or six membered carbocyclic ring, or
(iv) taken
together with R2 forms a heterocyclo optionally substituted with alkyl or
substituted alkyl;
BB is a bond, Ci_aalkylene, C2_4alkenylene, substituted Ci_aalkylene,
substituted C2-
4alkenylene, substituted C1_4alkylene-C(=9)NH-, -C(=3)NH-, -C1,4alkylene-
C(=0)NH-, -C(=0)NR19-, -C1_4alkylene-C(=3)NR19-, or substituted C1_4alkylene-
C(=0)NR19-, -(CHR14)6,-(CRI5R16)6- or -(CHRI4)p-C(=0)NH-;
R3a at each occurrence is selected independently from alkyl, substituted
alkyl,
halogen, haloalkoxy, cyano, nitro, keto, trifluoromethyl, -NRI7Ri 8, -S1217, -
0R17, -
SO2R17a, -SO2NR17R18, -NR17C(=0)R18, -0O2R17, --C(=0)R17, cyoloalkyl, aryl,
heterocyolo, and heteroaryl, wherein when R3a is cycloalkyl, aryl, heterocyclo
or heteroaryl,
said cycloalkyl, aryl, heterocyolo and heteroaryl in turn is optionally
substituted with alkyl or
substituted alkyl;
Z is a heteroaryl, for example an optionally-substituted bicyclic heteroaryl;
or
Z is triazolyl optionally substituted with one to two R7 substituents or
imidazolyl
optionally substituted with one to two R7 substituents and/or having fused
thereto a benzene
ring in turn optionally substituted with one to two R7 substituents; and
R7 is alkyl, carbamyl, or substituted alkyl;
R4 at each occurrence is selected independently of each other R4 from the
group
consisting of halogen, trifluoromethyl, OCF3, alkyl, substituted alkyl,
haloalkyl, nitro, cyano,
haloalkoxy, OR25, SR25, NR25R26, NR25S02R27, S02R27, S02NR25R26, CO2R26,
C(=0)R26,
C(=0)NR25R26, OC(=0)R25, -0C(=0)NR25R26, NR25C()R26, NR25CO2R26, aryl,
heteroaryl, heterocyclo and cycloalkyl;
R8 is Ci_aalkyl or phenyl optionally substituted with alkyl, halogen,
haloalkoxy, cyano,
nitro, or trifluoromethyl;
R9 is -NRioRii, alkyl, substituted alkyl, alkoxy, alkylthio, cycloalkyl, aryl,
heteroaryl, heterocyclo, or -0O2R12, alkyl or phenyl optionally substituted
with one to four
of halogen, cyano, trifluoromethyl, nitro, hydroxy, Ci_aalkoxy, haloalkoxy,
Ci_6alkyl,
CO2alkyl, SO2alkyl, SO2NH2, amino, NH(Ci_aalkyl), N(Ci_4alky1)2, NHC(=0)alky,
C(=0)alkyl, and/or Ci_aalkyl optionally substituted with one to three of
trifluoromethyl,
621
CA 3050553 2019-07-25
hydroxy, cyano, phenyl, pyridinyl; and/or a five or six membered heteroaryl or
heterocylo in
turn optionally substituted with keto or having a benzene ring fused thereto
or
a) Ci_aalkyl optionally substituted with one to two of:
i) SIZ13, 0R13, NRi3aRi3b, halogen, trifluoromethyl, CO2Ri3a, and
C(=34)NR13aRi3b;
ii) cycloalkyl optionally substituted with one to two of C(=C0)H, Ci4acyl,
alkenyl, carbamyl,
and/or phenyl in turn optionally substituted with halogen;
iii) phenyl or napthyl optionally substituted with one to two of halogen,
nitro, amino, alkyl,
hydroxy, Ci_aalkoxy, or having fused thereto a five or six membered
heterocyclo;
iv) pyridinyl, thiophenyl, furanyl, tetrahydrofuranyl, or azepinyl, optionally
substituted with
alkyl or having fused thereto a five to six membered carbocyclic ring
optionally substituted
with keto or Ci_4alkoxy;
b) 3 to 6 membered cycloalkyl optionally having up to four substituetits
selected from alkyl,
halogen, cyano, alkenyl, acyl, alkylthio, carbamyl, phenyl in turn optionally
substituted with
halogen; or having an aryl fused thereto;
c) pheyl optionally substituted with one to four of halogen, cyano,
trifluoromethyl, nitro,
hydroxy, Ci-aalkoxy, haloalkoxy, CI-4alkyl, CO2alkyl, SO2alkyl, SO2NH2, amino
NH(Ci-
4alkyl), N(Ci_4alky1)2, NHC(=3)alkyl, C(=0)alkyl, and/or Ci_aalkyl optionally
substituted
with one to three of trifluoromethyl, hydroxy, cyano, phenyl, pyridinyl;
and/or a five or six
membejed heteroaryl or heterocyle in turn optionally substituted with keto or
having a
benzene ring fused thereto;
d) pyridinyl, thiazolyl, furanyl, thiophenyl, and pyrrolyl optionally
substituted with one to
two of halogen, alkyl, and phenyl in turn optionally substituted with halogen
or
trifluoromethyl;
Rio and R11 are (i) independently selected from hydrogen, alkyl, substituted
alkyl,
alkoxy, heterocyclo, cycloalkyl, aryl, heteroaryl or Ci_aalkyl optionally
substituted with one
to two of¨0O2alkyl, ¨C(=3)NH(ary1), NH(ary1), cycloalkyl, phenyloxy, phenyl in
turn
optionally substituted with Ci_4alkyl, hydroxy, Ci_aalkoxy, halogen, amino,
nitro,
tetrahydrofuranyl, and/or five or six membered heterocyclo, or having a five
or six membered
heterocyclo fused thereto; pyrrolidinyl optionally substituted with keto;
napthyl, anthracenyl,
pyridinyl, thiophenyl, furanyl, imidazolyl, benzimidazolyl, or indolyl in turn
optionally
substituted with Ci_aalkyl or Ci_aalkoxy; or
(ii) taken together form a heteroaryl or heterocyclo selected from
pyrrolidinyl, piperazinyl,
piperidinyl, morpholinyl, tetrahydropyridinyl, and imidazoilidinyl, wherein
said heterocyclo
formed by Rio and R11 is optionally substituted with one to two of keto, CO2H,
Ciaalkoxy,
622
CA 3050553 2019-07-25
CO2alkyl, Ci_acarbamyl, benzyl; phenyl in turn optionally substituted with
alkyl, halogen, or
C14 alkoxy; tetrahydropyridinyl in turn optionally substituted with keto
and/or phenyl; alkyl
optionally substituted with amino or NHR21 wherein R21 is alkyl or phenyl
optionally
substituted with alkyl; and/or has a benzene ring fused thereto in turn
optionally substituted
with one to two of alkyl, Ci_4alkoxy, CO2alkyl, and/or C1.4carbamyl;
R12and R19 are hydrogen or alkyl;
R13 is hydrogen or alkyl;
R13a and R13b are selected from hydrogen, alkyl, and aryl;
R14, R15 and R16 at each occurrence are independently selected from hydrogen,
alkyl,
hydroxy, hydroxyCi_aalkyl, Ci_aalkoxy, and phenyl, and/or one of R15 and one
of Ri6join
together to form a 3 to 6 membered cycloalkyl;
Ri7and Risare independently selected from hydrogen, alkyl, substituted alkyl,
aryl,
phenyl, or benzyl wherein the phenyl or benzyl is optionally substituted with
alkyl, hydroxy,
or hydroxyalkyl;
Ri7a is alkyl or substituted alkyl;
R25 and R26 are independently selected from hydrogen, alkyl, or substituted
alkyl, or
taken together form a heterocyclo or heteroaryl ring;
R27 is alkyl or substituted alkyl;
q is 0, 1, 2, or 3;
m and n are 0, 1 or 2; and
p is 0, 1, 2, or 3.
[18] A method according to Claim 17, wherein L is hydrogen.
[19] A method according to Claim 17, wherein the S-enantiomer is in
enantiomeric excess.
[20] A method according to Claim 19, wherein the enantiomeric excess of S-
enantiomer
exceeds 70%.
[21] A method according to Claim 17, using the structure
623
CA 3050553 2019-07-25
...../ N
N/'''....
.¨..--N
N 0
CI s µµµ11 A
S N NH
H
CI
0
Cl
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
the enantiomeric excess (ee) of the S stereoisomer exceeds 70%.
[22] A method of treating, ameliorating, preventing, reversing or combating a
disease or
disorder, or unwanted/undesirable physiological process or its consequences or
an
unwanted/undesirable aesthetic, in a subject, selected from
(i) cancer, any cancer, neoplasia, metastasis, tumor
formation/growth/implantation,
tumorigenesis, solid tumor, blood borne tumor, cancer that is refractory or
resistant to
conventional chemotherapy, drug resistant tumor, multidrug resistant cancer;
(ii) cancer that metabolizes much of its glucose and/or glutamine to lactate,
for
example a cancer exhibiting the Warburg effect and/or a cancer that can be
discriminated
from surrounding tissue by PET imaging (e.g. '8F-FDG PET);
(iii) cachexia or cancer driven cachexia;
wherein the method comprises administering to the subject an effective amount
of at
least one compound, or a composition containing at least one compound, of the
following
formula:
624
CA 3050553 2019-07-25
RB
I
...,õ-N-..,...
N/
NN
).., ....õ,..
RAi ..,
N NL.. NH
H I
RA2
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof,
wherein:
RAI and RA2 are each independently selected from the groups
RD le' ___________
__________________________________________________
and
wherein Rc and RD are each independently selected from hydrogen, deuterium,
halogen and
alkyl, and wherein RE is hydrogen, deuterium, or alkyl;
RB is selected from RBI, hydrogen and deuterium;
wherein RBI is selected from phenyl, benzyl, pyridyl, pyrimidyl and pyrazinyl
optionally substituted with one or more substituents RB2;
wherein each RB2 is independently selected from halogen, alkyl, alkoxy, nitro,
amino,
methoxy and polyhalogen alkyl;
or RB is a phenylalkyl of the formula:
11 G (RG)q
(RF)q ----(C )n 1
wherein RF and RG are hydrogen or alkyl, G is a carbon-carbon double bond or a
carbon-
carbon single bond, n is 0 or 1 and q is 0 or 1 provided that where q is 0, G
is a carbon-carbon
double bond and where q is 1, G is a carbon-carbon single bond,
or RB is a diphenylalkyl of the formula
625
CA 3050553 2019-07-25
RH
RH
wherein RH is hydrogen or halogen, and p is 0, 1 or 2;
or RB is the group
Rj
41/
RK
wherein R and RK each independently represent 1-5 optional substituents on
each ring, and
wherein each fe and each RK, when present, is independently selected from
halogen, alkyl,
alkoxy, nitro, amino and polyhalogen alkyl.
1231 A method according to Claim 22, wherein RB is the group:
626
CA 3050553 2019-07-25
11)
RK
[24] A compound or composition for use according to Claim 22, wherein RB is
the group:
RL,
RM
wherein RL and Rm are each independently selected from halogen, alkyl, alkoxy,
nitro, amino
and polyhalogen alkyl.
[25] A method according to Claim 24, wherein RL and Rm are each independently
selected
from halogen.
[26] A method according to Claim 24, wherein RL and Rm are the same.
[27] A method according to Claim 24, wherein RL and Rm are each F.
627
CA 3050553 2019-07-25
[28] A method according to Claim 22, wherein RAI and RA2 are each
independently selected
from the group
.r\ ______________________________________ R C
RD
wherein Rc and RD are each independently selected from hydrogen, deuterium,
halogen and
alkyl.
[29] A method according to Claim 28, wherein RAI and RA2 are the same.
[30] A method according to Claim 28, wherein Rc is hydrogen.
[31] A method according to Claim 28, wherein RD is hydrogen.
[32] A method according to Claim 22, wherein the compound is:
FUF
N N
I I
CH2
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof.
[33] A method according to Claim 22, wherein the compound is an
isotopologue(s) of:
628
CA 3050553 2019-07-25
N
N N
H2C CH2
or a pharmaceutically-acceptable salt, solvate, hydrate or prodrug thereof.
[34] A method of treating, ameliorating, preventing, reversing or combating a
disease or
disorder, or unwanted/undesirable physiological process or its consequences or
an
unwanted/undesirable aesthetic, in a subject, selected from
(i) cancer that metabolizes much of its glucose and/or glutamine to lactate,
for
example a cancer exhibiting the Warburg effect and/or a cancer that can be
discriminated
from surrounding tissue by PET imaging (e.g. 18F-FDG PET);
(ii) cachexia, cancer driven cachexia, cancer fatigue, weight loss, weight
loss for
known or unknown reason, chronic wasting disease, atrophy, brown atrophy,
frailty, frailty
syndrome, aging frailty, geriatric syndrome, age-related cachexia and/or
sarcopenia,
weakness, wasting, anorexia nervosa, bulimia nervosa, eating disorder,
amenorrhea,
underweight, low body mass index (BMI, e.g. <18.5), low body fat percentage,
body
composition change, wasting syndrome, HIV wasting syndrome, malnutrition,
clinical
malnutrition, starvation, kwashiorkor syndrome, marasmus syndrome,
malabsorption,
malabsorption due to parasitic/bacterial infection (e.g. helminthiasis,
Whipple's disease, small
intestine bacterial overgrowth (SIB0), giardiasis etc.), anemia, refeeding
syndrome, appetite
loss, catabolysis, muscle atrophy, asthenia, muscle weakness (myasthenia),
sarcopenia,
osteoporosis, cachexia associated with HIV, AIDS, multiple sclerosis,
rheumatoid arthritis,
familial amyloid polyneuropathy, chronic kidney disease, cystic fibrosis,
multiple sclerosis,
motor neuron disease, Parkinson's disease, dementia, Addison's disease,
mercury poisoning
(acrodynia), chronic pancreatitis, untreated/severe type 1 diabetes mellitus,
hormonal
deficiency, tuberculosis, gastroenteritis, diarrhea, dysentery, any digestive
disease or
disorder, any gastrointestinal disease or disorder including functional
gastrointestinal
629
CA 3050553 2019-07-25
disorders, coeliac disease, tropical sprue, irritable bowel syndrome,
inflammatory bowel
disease, Crohn's disease, ulcerative colitis, short bowl syndrome, congestive
heart failure,
constrictive pericarditis, bradycardia, chronic obstructive pulmonary disease
(COPD),
altitude sickness, hyperthyroidism (subclinical hyperthyroidism, Graves'
disease,
multinodular goiter, toxic adenoma, inflammation of the thyroid {thyroiditis},
pituitary
adenoma), fatigue, chronic fatigue syndrome or any disease or disorder or
pathology in which
a body tissue(s) is undersupplied or underutilises (vs. its need) an
energetic/chemical
substrate(s), including 02;
(iii) cancer associated fever, which is especially associated with, but not
limited to,
non Hodgkin lymphoma (NHL), Hodgkin lymphoma, acute leukaemia, kidney cancer
(renal
cell cancer), liver cancer (hepatocellular carcinoma), bone cancer, adrenal
gland tumours
such as phaeochromocytomas, tumours in the hypothalamus, solid tumours;
(iv) disease or disorder or physiological process or condition that causes a
higher than
normal body temperature such as (without limitation) high environmental
temperature,
ingesting an uncoupler (e.g. 2,4-dinitrophenol), infection, sepsis,
neutropenic sepsis, stroke,
fever, pyrexia, hyperpyrexia, hyperthermia, malignant hyperthermia,
neuroleptic malignant
syndrome, serotonin syndrome, toxic serotonin syndrome, thyroid storm,
heatstroke, surgery
related, menopause ("hot flushes"), infection (non-limiting e.g. roseola,
measles, enteroviral
infections, parasitic, viral, fungal, Chlamydial, Rickettsia', bacterial,
mycobacterial, systemic
bacterial, intravascular, HIV associated, nosocomial), pyrogenic infection,
thermoregulatory
disorder(s), connective tissue disease(s), Kawasaki syndrome, drug overdose,
drug or drug
withdrawal induced hyperthermia, alcohol/drug withdrawal, idiosyncratic drug
reaction, fever
of known or unknown or uncertain origin (non-limiting e.g. infectious
disease(s),
inflammation, immunological disease(s), non-infectious inflammatory disease(s)
{non-
limiting eg. systemic rheumatic and autoimmune diseases, vasculitis,
granulomatous diseases,
autoinflammatory syndromes}, tissue destruction, reaction to incompatible
blood product(s),
metabolic disorder(s), inherited metabolic disorder(s), cancer, neoplasm,
endogenous or
exogenous pyrogen(s), injury, head injury);
(v) disease/disorder/injury/pathology/surgery
treatable/ameliorated/prevented/combated/helped by conferring hypothermia in a
subject for
some medical or other purpose which can include slowing a chemical reaction(s)
rate in a
subject for therapeutic benefit, preventing/minimizing brain and/or tissue
damage, slowing
physiological/pathological processes (reaction rates are temperature
dependent) and so
"buying time" to get the subject to treatment for their emergency (e.g.
trauma/septic shock or
630
CA 3050553 2019-07-25
other medical emergency), slowing the progress of sepsis until a sufficient
concentration of a
working antibiotic(s) can be built up in the subject (furthermore hypothermia,
by slowing
sepsis progression, buys time to observe which antibiotic(s) can work,
yielding time to try
alternative further antibiotic option(s) if required), used soon after or just
before clinical/legal
death to preserve the subject's organs/tissues until the subject can be
frozen/cryogenically
frozen or the pathology that caused clinical/legal death (e.g. wound) can be
fixed and the
subject resuscitated, administered to a subject when a first responder (e.g.
ambulance crew)
deems the subject dead or unlikely to survive the journey to a medical
facility (e.g. hospital)
wherein this administration helps to preserve the subject which is helpful if
hospital staff
subsequently assess that they can, or might be able to, save the subject,
stabilizing
surgical/trauma/Emergency Room (ER) patients, deep hypothermic circulatory
arrest for
surgery (DHCA, non-limiting applications of DHCA include repairs of the aortic
arch, repairs
to head and neck great vessels, repair of large cerebral aneurysms, repair of
cerebral
arteriovenous malformations, pulmonary thromboendarterectomy, resection of
tumors that
have invaded the vena cava, brain tumor resection {wherein the anti-cancer
activity of a
compound(s) of this invention juxtaposes well}), Emergency Preservation and
Resuscitation
(EPR), hypothermia for a surgical purpose, protective hypothermia during
surgery and/or
surgery complication, hypothermia to slow/reduce blood loss, hypothermia for
neuro- and/or
cardio- and/or organ/tissue and/or life protection in a subject that has
trauma/brain
trauma/polytrauma/surgery/stroke/ischemic stroke/hemorrhagic stroke/cardiac
arrest/myocardial infarction/hypoxia/shock (including, without limitation, low
volume,
cardiogenic, obstructive, and distributive shock)/sepsis/septic shock/multiple
organ
dysfunction syndrome/systemic inflammatory response syndrome (SIRS)/organ
failure/cytokine storm/anaphylactic shock/seizure/disseminated intravascular
.. coagulation/blocked
airway/croup/rhabdomyolysis/[head/facial/spinal/chest/abdominal/ballistic/knife
injury/trauma], or some other medical
emergency/condition/disorder/disease/injury/operation,
hypothermia for cardiac and/or cardiovascular surgery and/or open heart
surgery and/or brain
surgery (neurosurgery) and/or surgery using total circulatory arrest and/or
surgery using
cardiopulmonary bypass, Emergency Preservation and Resuscitation (EPR),
preserving
detached body parts such as limbs and/or organs (for example during organ
storage/transport
and/or transplant, thus increasing the time window for transplantation of
organs to recipients;
compound(s) of this invention is administered to organ to be transplanted [by
administration
to donor and/or by administration to isolated organ] and/or to organ
recipient, optionally
631
CA 3050553 2019-07-25
during transplant operation), protective hypothermia, targeted temperature
management,
therapeutic hypothermia, hypothermia therapy for neonatal encephalopathy,
neonatal
hypoxia-ischemia, birth asphyxia, hypoxic-ischemic encephalopathy (HIE),
haemorrhage,
hypovolemia, exsanguination, suspended animation, decompression sickness, burn
injury(s)
including skin burn, inflammation, allergic reaction, anaphylaxis,
tissue/organ rejection,
hypoxia, hypoxemia, anoxemia, anoxia, anemia, hypervolemia, altitude sickness,
obstructed
airway, asthma attack, hypoxia in a body/tissue/organ, hypoglycemia,
reperfusion injury
(ischemia-reperfusion injury), upon release of a ligature or tourniquet,
uraemia, crush
syndrome, compartment syndrome, traumatic brain and/or spinal cord injury,
major trauma,
infection, bacterial and/or viral infection(s) (non-limiting e.g. meningitis),
sepsis, septic
shock, stroke, cerebrovascular disease, ischemic brain injury, ischemic
stroke, traumatic
injury, brain injury, spinal cord injury, cardiac arrest, heart failure,
congestive heart failure,
Dilated cardiomyopathy, valvular heart disease, pulmonary embolism, adrenal
crisis,
Addisonian crisis, hypertensive emergency, haemorrhagic (hypovolemic) shock,
cardiogenic
shock, neurogenic shock, hepatic encephalopathy, blood loss, ischemic
brain/heart/kidney/intestinal injury, neuroprotection and/or card ioprotection
and/or tissue
protection during/after a stroke and/or ischemia and/or cardiac arrest and/or
resuscitation
and/or a period(s) of poor blood flow anywhere in a subject;
(vi) poisoning by a toxic amount of a compound(s) in a subject (non-limiting
e.g.
carbon monoxide/methanol/heavy metal/ethylene glycol/pesticide poisoning,
snake/spider/bee/insect/lizard venom, metabolic poison(s), nerve agent,
chemical weapon,
bacterial toxin(s) (e.g. food poisoning, Salmonella poisoning), endotoxemia,
eukaryote
produced toxin(s) e.g. (non-limiting) brevetoxin, drug(s)/substance(s)
overdose e.g. (non-
limiting) heroin, ethanol, a prescription medication(s), an over the counter
medication(s) such
as aspirin, paracetamol etc.; hypothermia is protective to toxic insult);
(vii) hypermetabolism (optionally because of one or more of, without
restriction,
traumatic brain injury, injury to the body, infection, sepsis, burn, multiple
trauma, fever,
long-bone fracture, hyperthyroidism, prolonged steroid therapy, surgery, bone
marrow
transplant, recovery from anorexia/bulimia), heat intolerence, insomnia, fatal
insomnia,
nervousness, Luft's disease, non-thyroidal hypermetabolism, thyrotoxicosis,
hyperthyroidism,
overactive thyroid, subclinical hyperthyroidism, too much thyroid hormone(s)
in the subject,
too much triiodothyronine (T3) and/or thyroxine (T4) in the subject,
hyperthyroxinemia
(including, without restriction, familial dysalbuminemic hyperthyroxinemia,
familial
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euthyroid hyperthyroxinemia, thyroid hormone resistance syndrome), thyroid
storm,
hyperthyroidism caused by one or more of (without restriction) Graves'
disease, thyroiditis,
Hashimoto's thyroiditis, subacute thyroiditis, postpartum thyroiditis, lumps
(nodules) on the
thyroid, enlarged thyroid gland (goitre), simple goitre, multinodular goiter,
toxic multinodular
goiter, toxic adenoma, toxic thyroid adenoma, inflammation of the thyroid,
hyperplasia of
thyroid, metastatic thyroid cancer, thyroid tumour, thyroid cancer (including,
without
restriction, papillary carcinoma, follicular carcinoma, medullary thyroid
carcinoma,
anaplastic thyroid carcinoma), eating too much iodine, goitrogen ingestion,
consumption of
ground beef contaminated with thyroid tissue ("hamburger hyperthyroidism"),
too much
synthetic thyroid hormone in the subject, pituitary adenoma, drug induced,
Amiodarone drug
induced, struma ovarii, Jod-Basedow syndrome, nonautoimmune autosomal dominant
hyperthyroidism;
(viii) low or less than desired metabolic/bioenergetic efficiency in a
subject, or low or
less than desired physical or mental performance (e.g. memory, IQ), or low or
less than
.. desired body weight, or fatigue/tiredness/weakness/exhaustion;
(ix) accelerated aging disease or progeroid syndrome including, without
restriction,
Werner syndrome, Bloom syndrome, De Barsy syndrome, Rothmund¨Thomson syndrome,
Cockayne syndrome, xeroderma pigmentosum, trichothiodystrophy, combined
xeroderma
pigmentosum-Cockayne syndrome, restrictive dermopathy, Wiedemann¨Rautenstrauch
syndrome, Hutchinson¨Gilford progeria syndrome (progeria), Ataxia
telangiectasia-like
disorder 2, XFE progeroid syndrome, Muscular dystrophy, Muscular Dystrophy
(Becker's,
Duchenne, Limb-Girdle), Yamamoto's Muscular Dystrophy, Mandibuloacral
dysplasia,
Dilated cardiomyopathy, GAPO syndrome, Cutis laxia, Ehlers-Danlos syndrom,
Lenz-
Majewski hyperostatic dwarfism, SHORT syndrome, Progressive external
opthalmoplegia,
Nester-Guillermo progeria syndrome, MDPL syndrome, Dyskeratosis congenital,
Down
syndrome;
(x) disease or disorder of aging (incidence/severity increases with increased
age/senescence) and/or unwanted/undesirable aspect(s) of aging and/or a
disease/disorder
associated with elevated reactive oxygen species including age-associated
decline, aging
frailty, frailty syndrome, sarcopenia, muscle weakness, osteoporosis,
cognitive decline,
cognitive defecit, mild cognitive impairment, degenerative diseases,
neurodegenerative
diseases, motor-associated neurodegenerative diseases, motor neuron disease,
amyotrophic
lateral sclerosis (ALS), primary lateral sclerosis, progressive muscular
atrophy, progressive
bulbar palsy, progressive supranuclear palsy, pseudobulbar palsy, hereditary
spastic
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CA 3050553 2019-07-25
paraplegia, Parkinson's disease, Multiple System Atrophy (MSA), Progressive
Supranuclear
Palsy (PSP), essential tremor, resting tremor, Alzheimer's disease,
Huntington's disease,
spinocerebellar ataxias, Friedreich's ataxia, dementia, frontotemporal
dementia, chronic
traumatic encephalopathy, memory loss, aged cognition, age/aging related
cognitive
decline/impairment, Batten disease, polyglutamine diseases, osteoporosis,
atherosclerosis,
cardiovascular disease, myocardial infarction, cerebrovascular disease,
stroke, heart failure,
heart failure with preserved ejection fraction, idiopathic pulmonary fibrosis,
fibrotic disease,
pulmonary disease, coronary artery disease, hypercholesterolemia, obesity,
liver disease, fatty
liver disease, lysosomal storage disease, amyloidosis, systemic sclerosis,
kidney disease,
hepatic cirrhosis, immunosenscence, clonal hematopoiesis, chronic obstructive
pulmonary
disease (COPD), hypertension, hypercholesterolemia, age-related thymic
atrophy, arthritis,
osteoarthritis, arthritis (Osteo-and Rheumatoid), Juvenile Rheumatoid
Arthritis (JRA),
Degenerative Disc Disease, Tendinopathy, Androgenetic Alopecia, male-pattern
baldness,
Idiopathic Pulmonary Fibrosis, systemic sclerosis, Chronic Obstructive
Pulmonary Disease,
Psoriasis, age-related loss of cardiac/pulmonary/cognitive/vision function,
diabetes, type 2
diabetes, andropause, glaucoma, retinal degeneration, sarcopenia, cachexia,
age-related
cachexia and/or sarcopenia, age-related macular degeneration (AMD,
early/intermediate/late), neovascular/wet AMD, dry AMD, Geographic atrophy
(GA), wet
and dry AMD in the same eye(s), Stargardt's macular degeneration, Best
vitelliform macular
dystrophy, diabetic retinopathy, proliferative diabetic retinopathy, diabetic
macular edema,
age/aging-related eye disease, ophthalmological disease/disorder, ocular
disease, vision loss,
progressive vision impairment, myopia (short-sightedness), degenerative
myopia, hyperopia
(far-sightedness), accommodative dysfunction, glaucoma, cataract formation,
retinal
degeneration, progressive retinal degeneration, retinitis pigmentosa, leber
hereditary optic
neuropathy, Fuchs spot, Best's disease, Sorsby's fundus dystrophy, hearing
loss (e.g. age-
related), presbycusis, tinnitus, naive T cell shortage, movement disability,
nonalcoholic fatty
liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), immunosenescence,
respiratory/urinary tract infection (RTI/UTI) especially in older/aged/elderly
subjects, cancer;
(xi) aging and/or one or more signs of aging, wherein one or more of these
compounds slow/delay/reduce/treat/prevent/stop/reverse aging, and/or extend
lifespan and/or
healthspan, and/or treat or delay the onset of geriatric aging of the
human/animal body,
tissue(s), or organ(s), and/or treat or delay the onset of an age-associated
phenotype in a
cell(s)/organism(s), and/or prolong fertility e.g. female fertility, delay
menopause;
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(xii) skin aging and/or damage (including sun damage) and/or scalp and/or hair
aging
and/or hair greying and/or hair loss;
(xiii) insomnia, fatal insomnia, sleep onset latency, delayed sleep phase
disorder,
exploding head syndrome, parasomnia, sleep-maintenance insomnia, sleep
disorder, too
much/inappropriate/undesired signals/activity/electrical activity in the
nervous system,
hyperactivity, hypersensitivity, Premature ejaculation, hyperreflexia,
Autonomic dysreflexia
(AD), Hyperventilation syndrome, brain hyperactivity, overly sensitive sensory
system,
pathological crying and/or laughing, Pseudobulbar affect (PBA, emotional
lability),
photophobia, phonophobia, temperature-sensitive, pressure-sensitive, brain
hyperexcitability,
overstimulation, intrusive thought(s), Perseveration, sensory overload,
disorganized thinking,
fantasy prone personality, malapdative daydreaming, dissociation, hyperkinetic
disorder,
agitation, Psychomotor agitation, restlessness, difficulty controlling
behaviour, disruptive
behaviour disorder, Emotional and behavioral disorder, pervasive developmental
disorder,
Overactive disorder associated with mental retardation and stereotyped
movements, attention-
deficit disorder, Attention Deficit Hyperactivity Disorder (ADHD), adult
attention-deficit
hyperactivity disorder, severe behavioral problem(s) in children (e.g., to
illustrate and not
restrict, combativeness and/or explosive hyperexcitable behavior {out of
proportion to
immediate provocation[s] }, hyperactive children who show excessive motor
activity with
accompanying conduct disorders consisting of one or more of: impulsivity,
difficulty
sustaining attention, aggressivity, mood lability, poor frustration
tolerance), Premenstrual
dysphoric disorder (PMDD), premenstrual syndrome (PMS), impulsiveness,
impulsivity,
impulse control disorder, lack of self-control, hysteria, histrionic
personality disorder,
attention difficulty, inattention, poor attention control, anxiety, paranoid
anxiety, Paranoid
personality disorder, distress, dysphoria, Adjustment disorder, separation
anxiety, anxiety
disorder, depressive anxiety, agitated depression, treatment-resistant
depression, Generalized
anxiety disorder, social anxiety disorder, stranger anxiety, separation
anxiety (e.g. in dogs left
at home), separation anxiety disorder, Mixed anxiety-depressive disorder,
depression (all
forms, all severities), restlessness/apprehension/anxiety before surgery.
hypochondria, panic
disorder, panic attack, emotional outburst, emotional instability,
Intermittent explosive
disorder, unreasonable/unwarranted anger/aggression, hyper-aggression,
hostility, rage, poor
temper control, self-hatred, poor attentional control, worry, irritability,
neuroses, somatization
disorder, somatic symptom disorder, pain disorder, psychological pain,
psychogenic pain,
psychogenic facial pain, Atypical odontalgia (AO), burning mouth syndrome,
throbbing,
toothache/pulpitis/dental pain, chronic lower back pain, negative emotion,
persistent/enduring
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negative emotion, body dysmorphic disorder, factitious disorder, illness
anxiety disorder,
unwarrented fight-or-flight response, stress, emotional stress, emotional
dysregulation,
distress, psychological stress, acute stress, chronic stress, acute stress
reaction, combat stress
reaction, traumatic grief, grief, grief after death of loved one, Prolonged
grief disorder (PGD),
heartbreak, guilt, shame, remorse, emotional pain, Algopsychalia, psychalgia,
suffering,
emotional trauma, psychological trauma, broken heart, Post Traumatic Stress
Disorder
(PTSD), Complex post-traumatic stress disorder (C-PTSD), hypervigilance,
sympathetic
hyperactivity, inability or impaired ability to relax, flashbacks, dysphoric
hyperarousal,
agoraphobia, insomnia, fatal insomnia, mood disorder, irrational/unwarrented
fear/terror,
phobia, social phobia, Cancerophobia, thunderstorm/firework phobia,
hypersexuality,
hypersexual disorder, depression, clinical depression, unipolar depression,
bipolar disorder,
Bipolar I, Bipolar II, Bipolar disorder not otherwise specified (Bipolar NOS),
cyclothymia,
cyclothymic disorder, mixed affective state, atypical depression, melancholic
depression,
postpartum depression, double depression, seasonal affective disorder, mania,
manic episode,
hypomania, increase in energy of psychomotor activity, delirium, excited
delirium, major
depressive disorder, minor depressive disorder, recurrent brief depression,
Depressive
Disorder Not Otherwise Specified (DD-NOS), major depressive episode,
persistent
depressive disorder (PDD), dysthymia, dysthymic disorder, absence of euthymia,
manic
thoughts, racing thoughts, thought disorder, disordered thinking, reduced
ability to plan and
.. execute tasks, paranoia, hallucination (including, without limitation,
visual, auditory,
olfactory, gustatory, tactile, proprioceptive, equilibrioceptive, nociceptive,
thermoceptive,
chronoceptive), Charles Bonnet syndrome, delusion, persecutory delusion,
hearing voices,
homicidal/criminal ideation/tendency/thoughts, suicidal
ideation/tendancy/thoughts, self-
injury, non-suicidal self-injury, violence, attacking others, negative mood
swing, personality
disorder, Borderline personality disorder, Narcissistic personality disorder,
malignant
narcissism, dissociative disorder, dissociative identity disorder (DID),
Psychosis, acute
psychosis, chronic psychosis, psychosis spectrum disorder, manifestations of
psychotic
disorders, behavioral complications of mental retardation, stimulant
psychosis, psychotic
depression, hallucinogen persisting perception disorder, Psychoactive
substance-related
disorder, amphetamine-induced psychosis, brief psychotic disorder, Brief
reactive psychosis,
Menstrual psychosis, postpartum psychosis, Psychotic disorder, Psychopathy,
chronic
hallucinatory psychosis, manifestation(s) of psychotic disorder,
neurotic/reactive/endogenous/involutional/psychotic depression/depressive
disorder
(optionally accompanied by anxiety or agitation), depressive neurosis,
delusional depression,
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CA 3050553 2019-07-25
psychotic aggression, psychiatric symptoms of dementia, AIDS delirium,
Supersensitivity
psychosis, Tardive psychosis, Tardive dysmentia, Depersonalization disorder,
out-of-body
experience, Sociopathy, Schizophrenia, Paranoid schizophrenia, disorganized-
type
schizophrenia, simple-type schizophrenia, pseudoneurotic schizophrenia,
prodromal
schizophrenia, schizoaffective disorder, bipolar type schizoaffective
disorder, depressive type
schizoaffective disorder, schizoaffective psychosis, Schizotypal personality
disorder,
schizophreniform disorder, Delusional parasitosis, formication, paresthesias,
Acroparesthesia,
tinnitus, delusional disorder, delusional jealousy, inappropriate behaviour,
behavioural
disorder, antisocial personality disorder, Oppositional defiant disorder
(ODD), conduct
disorder (CD), Disruptive mood dysregulation disorder (DMDD), sadistic
personality
disorder, poor inhibitory control, kleptomania, Pyromania, trichotillomania,
dermatillomania,
pathological/problem gambling, dyskinesia, tardive dyskinesia, paroxysmal
dyskinesia,
Paroxysmal kinesigenic dyskinesia, Paroxysmal nonkinesigenic dyskinesia,
Paroxysmal
exercise-induced dystonia, Hemiballismus, tic disorder, tremor, clonus,
Tourette's syndrome,
coprolalia, copropraxia, Echophenomena, Echopraxia, Echolalia, Palilalia,
stuttering/stammering, stereotypy, punding, Self-stimulatory behaviour
(stimming),
Stereotypic movement disorder (SMD), synesthesia, obsession,
Obsessive¨compulsive
disorder (OCD), obsessive¨compulsive personality disorder, anankastic
personality disorder,
relationship obsessive¨compulsive disorder (ROCD), Scrupulosity, Primarily
obsessional
obsessive compulsive disorder, sexual obsession, Akathisia (including, without
limitation,
chronic, acute, Pseudoakathisia, Tardive akathisia, withdrawal or "rebound"
akathisia),
Restless legs syndrome, motor restlessness, periodic limb movement disorder
(PLMD),
periodic limb movements in sleep (PLMS), Sydenham's chorea, chorea, dystonia,
Hypnic
jerk, twitching, spasms, Tetanus, tetanus muscle spasms, tetanized state,
Myoclonus,
myoclonic seizure, Action myoclonus, Palatal myoclonus, Middle ear myoclonus,
Spinal
myoclonus, Stimulus-sensitive myoclonus, Sleep myoclonus, Cortical reflex
myoclonus,
Essential myoclonus, myoclonic epilepsy, Juvenile myoclonic epilepsy,
Progressive
myoclonus epilepsy (PME, including, without limitation, Dentatorubral-
pallidoluysian
atrophy, Unverricht-Lundborg disease, MERRF syndrome, Lafora disease),
Reticular reflex
myoclonus, Myoclonic epilepsy, diaphragmatic flutter, automatism, status
epilepticus,
Epilepsia partialis continua, Complex partial status epilepticus, epilepsy,
epileptic seizure,
simple partial seizure, complex partial seizure, generalized epilepsy,
generalized seizure
(including, without limitation, tonic-clonic, tonic, clonic, myoclonic,
absence (including
typical absence and atypical absence), atonic seizure), focal epilepsy, focal
seizure,
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CA 3050553 2019-07-25
focal/partial seizure (including, without limitation, Simple partial seizure
and Complex partial
seizure), focal aware seizure, focal impaired awareness seizure, generalised
epilepsy,
temporal lobe epilepsy (including, without restriction, mesial temporal lobe
epilepsy
{MTLE} and lateral temporal lobe epilepsy ILTLED, Frontal lobe epilepsy,
Rolandic
epilepsy, Nocturnal epilepsy, Nocturnal frontal lobe epilepsy, Autosomal
dominant nocturnal
frontal lobe epilepsy (ADNFLE), Generalized epilepsy with febrile seizures
plus (GEFS+),
Panayiotopoulos syndrome, epileptic fit, reflex epilepsy, reflex seizure,
absence seizure
(including, without limitation, childhood absence epilepsy, epilepsy with
myoclonic
absences, juvenile absence epilepsy, juvenile myoclonic epilepsy, Jeavons
syndrome {eyelid
myoclonia with absences}, genetic generalised epilepsy with phantom absences),
complex
partial seizure, atonic seizure, generalized tonic-clonic seizure,
tonic¨clonic seizure, extrinsic
stimulus epilepsy, intrinsic stimulus epilepsy, photosensitive epilepsy,
musicogenic epilepsy,
thinking epilepsy, eating epilepsy, seizure(s), Febrile seizure, nerve agent
induced seizure,
Dravet syndrome (sometimes modest hyperthermic stressors like physical
exertion or a hot
bath can provoke seizures in affected individuals), acute symptomatic seizure,
seizure-related
disorder, drug related seizure, paroxysmal depolarizing shift, Ohtahara
syndrome, Epilepsy in
females with mental retardation, Rasmussen's encephalitis, Epilepsy syndrome,
benign
rolandic epilepsy, childhood absence epilepsy, absence epilepsy, juvenile
myoclonic
epilepsy, epileptic encephalopathies, Lennox¨Gastaut syndrome, West syndrome
(Epileptic
.. spasms), Doose syndrome (Myoclonic astatic epilepsy, MAE), Lennox-Gestaut
syndrome,
pseudo-Lennox Gastaut syndrome (atypical benign partial epilepsy), Benign
familial neonatal
epilepsy, Benign occipital epilepsy of childhood, familial neonatal
convulsions, Febrile
infection-related epilepsy syndrome, interictal dysphoric disorder, euphoria
sclerotic,
psychogenic non-epileptic seizure, non-epileptic seizure, gelastic seizure,
convulsion(s),
migraine, status migrainosus, tension headache, headache, Hypnic headache,
hiccups,
intractable hiccups, thumps in equines, Postural orthostatic tachycardia
syndrome (POTS),
Pheochromocytoma, agony, pain, chronic pain, acute pain, pain due to
disease/injury,
neuropathic pain, fibromyalgia, postherpetic neuralgia, phantom pain, referred
pain, back
pain, lower back pain, pelvic pain, cancer associated pain, chemotherapy
associated pain,
.. Diabetic neuropathy, small fiber peripheral neuropathy, Mononeuritis
multiplex,
Wartenberg's migratory sensory neuropathy, Breakthrough pain, idiopathic pain,
polyneuropathy, Neuritis, Mononeuropathy, Polyradiculoneuropathy, Radial
neuropathy,
neuropathy, visceral pain, Burning feet syndrome, Tarsal tunnel syndrome,
Carpal tunnel
syndrome, Guyon's canal syndrome, Cubital Tunnel Syndrome, Repetitive strain
injury,
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CA 3050553 2019-07-25
Sciatica, Neurofibromatosis, Ulnar nerve entrapment, Erythromelalgia,
Paroxysmal extreme
pain disorder, Proctalgia fugax, dysesthesia, scalp dysesthesia, dysesthetic
burning,
hyperesthesia, hyperalgesia, Opioid-induced hyperalgesia, hyperpathia,
allodynia, pain
response from stimuli which do not normally provoke pain, Complex regional
pain syndrome
(said to be most painful condition known to man), Radiculopathy, neuralgia
(including,
without restriction, intercostal neuralgia, trigeminal neuralgia, atypical
trigeminal neuralgia,
glossopharyngeal neuralgia, occipital neuralgia, postherpetic neuralgia),
ciguatera poisoning,
irritable bowel syndrome (IBS), interstitial cystitis (IC), temporomandibular
joint disorder,
acute intermittent porphyria, Porphyria, Acute porphyria (including, without
limitation, acute
intermittent porphyria {AIP}, variegate porphyria {VP}, aminolevulinic acid
dehydratase
deficiency porphyria {ALA)}, hereditary coproporphyria {HCP}, drug induced),
Chronic
porphyria (including, without limitation, X-linked dominant protoporphyria
{XLDPP},
congenital erythropoietic porphyria {CEP}, porphyria cutanea tarda {PCT}, and
erythropoietic protoporphyria {EPP}), cutaneous porphyria, Porphyria cutanea
tarda, allergy,
allergic reaction, anaphylaxis, anaphylactic shock, hives, asthma, allergic
rhinitis, rhinitis,
urticaria, contact dermatitis, cough, sore throat, esophageal reflux disease,
heartburn, chest
pain, Esophageal motility disorder, Nutcracker esophagus, diseases involving
gastrointestinal
motility, Peptic ulcer disease, esophageal spasm, angina, itchiness, Pruritus,
severe pruritus,
Prurigo, Pruritic skin condition, chronic itch, eczema, pruritus in eczema,
neuropathic itch,
neurogenic itch, itchiness due to atopic dermatitis and/or lichen simplex
chronicus, peripheral
sensitization, central sensitization, sensory perception of absent stimuli,
too much sensory
stimulation, sensory stimulation brings discomfort, Neuromyotonia, Peripheral
nerve
hyperexcitability, Morvan's syndrome, Benign fasciculation syndrome, Cramp
fasciculation
syndrome, hyperhidrosis, undifferentiated somatoform disorder, somatoform
disorder,
somatic symptom disorder, conversion disorder, functional neurological symptom
disorder,
severe nausea and/or vomiting, severe nausea/emesis, Chemotherapy-induced
peripheral
neuropathy, chemotherapy-induced nausea and vomiting (CINV), radiation therapy-
induced
nausea and vomiting (RINV), postnarcotic nausea, hyperemesis gravidarum,
morning
sickness, Cyclic vomiting syndrome, retching/dry heaving, Urinary
incontinence, enuresis,
nocturnal enuresis, tail chasing, reduce urine spraying/marking behavior,
benzodiazepine
withdrawal syndrome, barbituate withdrawal syndrome, Neuroleptic
discontinuation
syndrome, drug withdrawal discomfort/pain/symptoms, drug use disorder, alcohol
use
disorder, opoid use disorder, amphetamine use disorder, cocaine use disorder,
alcohol
withdrawal syndrome/symptoms, delirium tremens, opoid withdrawal
sydrome/symptoms,
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CA 3050553 2019-07-25
drug craving, drug addiction, drug dependence, polysubstance dependence, drug
overdose,
smoking, tobacco (nicotine) addition, tobacco (nicotine) withdrawal symptoms,
alcoholism,
addiction, opoid addiction, cocaine/crack addiction, addictive behaviour,
addictive
personality, behavioural addiction, internet/computer/computer game/social
media/media
addiction, exercise addiction, compulsive behaviour (e.g. {non-limiting}
checking, counting,
washing, repeating), anti-social behaviour, criminality, sexual compulsion,
impulsive sexual
behaviour, compulsive buying, gambling addiction, sex related addiction,
sexual urge,
hunger, eating desire/compulsion, eating disorder, polyphagia, overeating,
binge eating
disorder, compulsive overeating, insatiable/excessive appetite, bulimia
nervosa, anorexia
nervosa, substance abuse, substance-induced delirium, substance-induced
psychosis,
substance-induced mood disorder, drug overdose, vertigo, motion sickness,
seasickness,
mental/nervous breakdown, Autism spectrum disorder, neurological disorder,
cognitive
disorder, mental disorder, mental health disorder, mental health condition
involving impaired
or altered neural plasticity, mood disorder, mental disorder disclosed in
Diagnostic and
Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) or a later
edition, a
mental/behavioural disorder disclosed by the International Classification of
Diseases (ICD) in
LCD-10 Chapter V: Mental and behavioural disorders (World Health Organisation,
WHO); or
(xiv) diseases or disorders or conditions or pathologies or
unwanted/undesirable
effects/actions/behaviour treatable/ameliorated/prevented/combated, in
totality or in part (e.g.
along with surgery), by anaesthesia, pre-anaesthesia, post-anaesthesia,
hypoesthesia,
hypoactivity, sedation, coma, tranquilization, behavioural submission, muscle
relaxation,
hibernation, artificial hibernation, torpor, synthetic torpor, suspended
animation (e.g. used
during spaceflight because e.g. reduces ionizing radiation damage to subject);
(xv) hyperproliferative/hyperplasia disorder, non-cancerous proliferative
disorder,
hyperproliferative autoimmune disorder, hyperplasia, epidermal hyperplasia,
dysplasia (e.g.
epithelial dysplasia), nodule(s), wart(s), papilloma(s), squamous cell
papilloma, genital
wart(s), condyloma(s), condyloma acuminatum, cyst(s), polyp(s) {including,
without
restriction, digestive, colorectal, endometrial, cervical, nasal, laryngeal,
inflammatory fibroid
polyp[s])}, inherited/hereditary (including, without restriction, Familial
adenomatous
polyposis, Peutz¨Jeghers syndrome, Turcot syndrome, Juvenile polyposis
syndrome, Cowden
disease, Bannayan¨Riley¨Ruvalcaba syndrome {Bannayan-Zonana syndrome},
Gardner's
syndrome) and non-inherited (non-restrictive e.g. Cronkhite¨Canada syndrome)
polyposis
syndrome, benign tumour, adenoma, organ enlargement by hyperplasia, Cushing's
disease
(enlarged adrenal cortex by hyperplasia), congenital adrenal hyperplasia,
hyperplasia of
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CA 3050553 2019-07-25
breast, atypical ductal hyperplasia, intraductal papillomatosis,
fibroadenomas, fibrocystic
changes, hemihyperplasia, focal epithelial hyperplasia, sebaceous hyperplasia,
sebaceous
adenoma, intimal hyperplasia, unwanted/undesirable smooth muscle cell
proliferation,
smooth muscle cell hyperplasia, intimal smooth muscle cell hyperplasia,
neointimal
hyperplasia, proliferative vascular disorders, stenosis, stenosis because of
cellular
proliferation, vaginal stenosis, stenosis in a blood vessel, lessoned patency
of a blood vessel,
stenosis in a blood vessel because of cellular proliferation, vascular
occlusion, restenosis,
restenosis in a blood vessel that has been implanted with a stent, in-stent
restenosis, post-
angioplasty restenosis, systemic sclerosis, cirrhosis of the liver, adult
respiratory distress
syndrome, idiopathic cardiomyopathy, lupus erythematosus, retinopathy (e.g.
diabetic
retinopathy and/or other retinopathy[y/ies]), cardiac hyperplasia, fibrosis,
pulmonary fibrosis,
idiopathic pulmonary fibrosis, fibromatosis, neurofibromatosis, renal
interstitial fibrosis,
Cowden syndrome, hamartoma(s), choristoma(s), hemangioma(s), lymphangioma(s),
rhabdomyoma(s), lymphangiomatosis, cystic hygroma, trichilemmoma, sarcoidosis,
neurosarcoidosis, aggressive fibromatosis, desmoid tumour(s),
unwanted/undesirable skin
cell proliferation, hyperproliferative skin disorder, psoriasis (including,
without restriction,
plaque, guttate, inverse, pustular, napkin, seborrheic-like, nail, scalp and
erythrodermic
psoriasis), psoriatic arthritis, dactylitis, seborrhoeic dermatitis, dandruff,
eczema, atopic
dermatitis, rosacea, reactive arthritis (Reiter's syndrome), pityriasis rubra
pilaris,
hyperproliferative variants of disorders of keratinization (e.g., without
restriction, actinic
keratosis, senile keratosis, keratosis pilaris, seborrheic keratosis),
scleroderma, benign
prostatic hyperplasia, prostate enlargement, endometrial hyperplasia, atypical
endometrial
hyperplasia, benign endometrial hyperplasia, adenomyosis, atypical polypoid
adenomyoma,
endometriosis, endometriosis of ovary (endometrioma), endometrial polyp(s),
polycystic
ovary syndrome, ovarian cyst(s), cervical polyp(s), uterine fibroid(s),
uterine hyperplasia;
(xvi) Tumour Associated Macrophages (TAMs) or any macrophage associated
disease or disorder such as, without limitation, Macrophage Activation
Syndrome (MAS),
HIV, AIDS, HIV-associated neurocognitive disorders (HAND), AIDS dementia, HIV
peripheral neuropathy, HIV associated cancers, AIDS-defining cancers, non-AIDS
defining
cancers, any disease in which the pathogen(s) hides from the immune system in
macrophages
including, without limitation, Mycobacterium tuberculosis (causes
tuberculosis), Leishmania
parasite (causes Leishmaniasis), Chikungunya virus (causes Chikungunya),
Legionella
pneumophila (causes Legionnaires' disease), adenoviruses, T. whipplei (causes
Whipple's
Disease), Brucella spp. (causes brucellosis), Staphylococcus aureus, Ebola
virus, Hepatitis B
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virus, Hepatitis C virus, influenza virus strains, dengue virus and antibiotic
resistant bacteria,
any disease or condition in which activated macrophages are unwanted or
undesirable;
(xvii) virus/pathogen neuroinvasion via macrophage(s), as used for non-
limiting
example by HIV, Heptatitis C virus and SARS coronavirus;
(xviii) neurocognitive or neurodegenerative diseases/disorders, for non-
limiting
example those caused by a virus;
(xix) virus/pathogen transmission from mother to fetus/baby via macrophage(s)
as
used for non-limiting example by zika (via Hofbauer cells) and HIV
(macrophages in breast
milk);
(xx) acute or chronic or systemic inflammation or any inflammatory
disease/disorder/syndrome or any autoinflammatory disease/disorder/syndrome or
any
autoimmune disease/disorder/syndrome;
(xxi) acute inflammation, chronic inflammation, systemic inflammation,
inflammation
because of infection or foreign bodies or injury or chemical or toxin or drug
or stress or
frostbite or burn or ionising radiation or surgery, inflammatory
diseases/disorders/syndromes,
Macrophage Activation Syndrome (MAS), autoinflammatory
diseases/disorders/syndromes,
age-related chronic inflammatory diseases ("inflammaging"), autoimmune
diseases/disorders/syndromes, diseases/disorders of the innate immune system,
sore throat,
sore throat associated with cold or flu or fever, high-intensity exercise
associated
inflammation, ulcerative colitis, inflammatory bowel disease (IBD), irritable
bowel syndrome
(IBS), rheumatoid arthritis, osteoarthritis, inflammatory osteoarthritis,
psoriatic arthritis,
atopic dermatitis, allergic airway inflammation, asthma, inflammation
associated depression,
neuroinflammation, neuropathic pain, exercise-induced acute inflammation,
atherosclerosis,
allergy, hay fever, anaphylaxis, inflammatory myopathies, drug-induced
inflammation,
systemic inflammatory response syndrome, sepsis-related multiple organ
dysfunction/multiple organ failure, microbial infection, acute
brain/lung/hepatic/renal
injuries, lung inflammation, acute lung injury (ARDS), acne vulgaris, celiac
disease, celiac
sprue, chronic prostatitis, colitis, autoimmune hemolytic anemia,
diverticulitis,
glomerulonephritis, proliferative glomerulonephritis, membranous nephropathy,
minimal
change nephrotic syndrome, hidradenitis suppurativa, hypersensitivities,
interstitial cystitis,
Mast Cell Activation Syndrome, mastocytosis, otitis, pelvic inflammatory
disease (PID),
endometritis, reperfusion injury, rheumatic fever, rhinitis, sarcoidosis,
transplant rejection,
parasitosis, eosinophilia, type III hypersensitivity, ischaemia, chronic
peptic ulcer,
tuberculosis, Crohn's disease, hepatitis, chronic active hepatitis, immune
hepatitis, alcoholic
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hepatitis, chronic viral hepatitis, ankylosing spondylitis, diverticulitis,
fibromyalgia, systemic
lupus erythematous (SLE), Alzheimer's disease, Parkinson's disease,
neurodegenerative
disease, cardiovascular disease, chronic obstructive pulmonary disease,
bronchitis, acute
bronchitis, bronchiectasis, bronchopneumonia, obliterative bronchiolitis,
appendicitis, acute
appendicitis, bursitis, cystitis, dermatitis, encephalitis, HIV encephalitis,
gingivitis,
meningitis, infective meningitis, myelitis, nephritis, neuritis,
periodontitis, chronic
periodontitis, phlebitis, prostatitis, RSD/CRPS, rhinitis, sinusitis, chronic
sinusitis, tendonitis,
testiculitis, tonsillitis, urethritis, vasculitis, respiratory
bronchiolitis¨associated interstitial
lung disease and desquamative interstitial pneumonia, pneumonia, interstitial
lung disease,
Lofgren syndrome, Heerfordt syndrome, monocytosis, liver fibrosis,
steatohepatitis,
nonalcoholic steatohepatitis, silicosis, histiocytoses, Langerhans' cell
histiocytosis,
haemophagocytic lymphohistiocytosis, pulmonary langerhans cell histiocytosis,
obesity, type
II diabetes, gout, pseudogout, organ transplant rejection, epidermal
hyperplasia, chronic
fatigue syndrome, graft versus host disease (GVHD), lymphadenopathy,
rheumatoid arthritis
(RA), osteoarthritis (OA), inflammatory osteoarthritis, lupus, multiple
sclerosis (MS),
myocarditis, uveitis, CNS disease(s), inflammation aspect to a CNS disease(s),
hypothalamic
inflammation, dementia, glaucoma, amyloid related/driven disease, lipid
storage disease(s),
fibrosis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic
steatohepatitis (NASH),
cirrhosis, cirrhosis of the liver, alcoholic cirrhosis, nephropath(y/ies),
lupus nephritis,
immune nephritis, fibrotic disorder(s), cardiovascular disease, heart disease,
atherosclerosis,
vulnerable plaque, plaque formation, lipid containing macrophage related
disease(s)/disorder(s)/malad(y/ies), macrophage foam cells, diabetes, type I
diabetes, type 2
diabetes, insulin resistance, macrophage aspect to insulin resistance,
obesity, obesity
associated inflammation, macrophage accumulation/large numbers of macrophages
in
adipose tissue (e.g. associated with obesity), granuloma(s), granulomatous
diseases,
sarcoidosis (including, without limitation, Annular sarcoidosis, Erythrodermic
sarcoidosis,
Ichthyosiform sarcoidosis, Hypopigmented sarcoidosis, Lofgren syndrome, Lupus
pemio,
Morpheaform sarcoidosis, Mucosal sarcoidosis, Neurosarcoidosis, Papular
sarcoid, Scar
sarcoid, Subcutaneous sarcoidosis, Systemic sarcoidosis, Ulcerative
sarcoidos),
neurosarcoidosis, pulmonary sarcoidosis, interstitial lung disease, pulmonary
fibrosis,
pulmonary tuberculosis, immune reconstitution syndrome of HIV,
Jarisch¨Herxheimer
reaction, sepsis, Paget's disease of bone, osteolysis, monocytosis,
histiocytosis, X-type
histiocytoses, non-X histiocytoses, Langerhans cell histiocytosis, non-
Langerhans-cell
histiocytosis, malignant histiocytosis, malignant histiocytic disorders,
histiocytomas,
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histiocytic lymphoma, hemophagocytic syndrome, hemophagocytic
lymphohistiocytosis,
lymphohistiocytosis, diffuse histiocytic sarcoma, Rosai¨Dorfman disease,
gliosis, Bergmann
gliosis, Chronic Obstructive Pulmonary Disease (COPD), chronic inflammatory
lung disease,
familial mediterranean fever (FMF), TNF receptor-associated periodic syndrome
(TRAPS),
Hyperimmunoglobulinemia D with recurrent fever syndrome (HIDS), cryopyrin
associated
periodic syndrome (CAPS), Blau syndrome, Majeed syndrome, deficiency of
interleukin-1
receptor antagonist (DIRA), mevalonate kinase deficiency, pyogenic-arthritis-
pyoderma
gangrenosum and acne syndrome (PAPA), periodic fever aphthous stomatitis
pharyngitis
adenitis (PFAPA) syndrome, Behcet's disease, Still's disease, Crohn's disease,
Schnitzler's
syndrome, Sweet's syndrome, NLRP12-associated autoinflammatory disorders,
deficiency of
interleukin-1 receptor antagonist (DIRA), pyoderma gangrenosum, cystic acne,
aseptic
arthritis, periodic Fever Associated with mevalonate kinase deficiency
(hyperimmunoglobulin D Syndrome), Pyogenic Arthritis Pyoderma Gangrenosum Acne
(PAPA) syndrome, Periodic Fever Aphthous Stomatitis, Pharyngitis and
Adenopathy
(PFAPA) syndrome, Adult-Onset Still's Disease (AOSD), Systemic Juvenile
Idiopathic
Arthritis (sJIA), Chronic Recurrent Multifocal Osteomyelitis (CRMO), Synovitis
Acne
Pustulosis Hyperostosis Osteitis (SAPHO) syndrome, Cryopyrin associated
Periodic
Syndrome (CAPS), Familial cold auto inflammatory syndrome (FCAS), Muckle-Wells
syndrome (MWS), Familial cold urticarial, Neonatal onset multisystemic
inflammatory
disorder (NOMID), hereditary Periodic Fever Syndromes, Periodic Fever
Syndromes,
systemic autoinflammatory diseases, Addison's disease, Agammaglobulinemia,
Alopecia
areata, Amyloidosis, Ankylosing spondylitis, Anti-GBM/Anti-TBM nephritis,
Antiphospholipid syndrome, autoimmune angioedema, autoimmune dysautonomia,
autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune inner ear
disease
(AIED), autoimmune myocarditis, autoimmune pancreatitis, autoimmune
retinopathy,
autoimmune urticaria, axonal & neuronal neuropathy (AMAN), Bak) disease,
Behcet's
disease, benign mucosal pemphigoid, Bullous pemphigoid, Castleman disease
(CD), Celiac
disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy
(CIDP),
chronic recurrent multifocal osteomyelitis (CRMO), Churg-Strauss, Cicatricial
pemphigoid,
Cogan's syndrome, cold agglutinin disease, congenital heart block, coxsackie
myocarditis,
CREST syndrome, Berger's disease, dermatitis herpetiformis, dermatomyositis,
Devic's
disease (neuromyelitis optica), discoid lupus, Dressler's syndrome,
endometriosis,
eosinophilic esophagitis (EoE), eosinophilic fasciitis, erythema nodosum,
essential mixed
cryoglobulinemia, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant
cell arteritis
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(temporal arteritis), giant cell myocarditis, glomerulonephritis,
proliferative
glomerulonephritis, membranous nephropathy, minimal change nephrotic syndrome,
Goodpasture's syndrome, granulomatosis with polyangiitis, Graves' disease,
Guillain-Barre
syndrome, Hashimoto's thyroiditis, hemolytic anemia, immune hemolytic anemia,
Henoch-
Schonlein purpura (HSP), Herpes gestationis or pemphigoid gestationis (PG),
hypogammalglobulinemia, IgA Nephropathy, IgG4-related sclerosing disease,
Immune
thrombocytopenic purpura (ITP), Inclusion body myositis (IBM), Interstitial
cystitis (IC),
juvenile arthritis, juvenile diabetes (Type I diabetes), juvenile myositis
(JM), Kawasaki
disease, Lambert-Eaton syndrome, Leukocytoclastic vasculitis, Lichen planus,
Lichen
sclerosus, Ligneous conjunctivitis, Linear IgA disease (LAD), Lupus, Lyme
disease chronic,
Meniere's disease, Microscopic polyangiitis (MPA), Mixed connective tissue
disease
(MCTD), Mooren's ulcer, Mucha-Habermann disease, Multiple sclerosis,
Myasthenia gravis,
Myositis, Narcolepsy, Neuromyelitis optica, Neutropenia, Ocular cicatricial
pemphigoid,
Optic neuritis, Palindromic rheumatism (PR) PANDAS, Paraneoplastic cerebellar
.. degeneration (PCD), Paroxysmal nocturnal hemoglobinuria (PNH), Parry
Romberg
syndrome, Pars planitis (peripheral uveitis), Parsonnage-Tumer syndrome,
Pemphigus,
peripheral neuropathy, perivenous encephalomyelitis, Pernicious anemia (PA),
POEMS
syndrome, Polyarteritis nodosa, Polyglandular syndromes type I, II, III,
Polymyalgia
rheumatica, Polymyositis, Postmyocardial infarction syndrome,
Postpericardiotomy
syndrome, Primary biliary cirrhosis, Primary sclerosing cholangitis,
Progesterone dermatitis,
Psoriasis, Psoriatic arthritis, Pure red cell aplasia (PRCA), Pyoderma
gangrenosum,
Raynaud's phenomenon, Reactive arthritis, Reflex sympathetic dystrophy,
Relapsing
polychondritis, Restless legs syndrome (RLS), Retroperitoneal fibrosis,
Rheumatic fever,
Rheumatoid arthritis, Sarcoidosis, Schmidt syndrome, Scleritis, Scleroderma,
Sjogren's
syndrome, Sperm & testicular autoimmunity, Stiff person syndrome (SPS),
Subacute
bacterial endocarditis (SBE), Susac's syndrome, Sympathetic ophthalmia (SO),
Takayasu's
arteritis, Temporal arteritis/Giant cell arteritis, Thrombocytopenic purpura
(TTP), Tolosa-
Hunt syndrome (THS), Transverse myelitis, Type I diabetes, Ulcerative colitis
(UC),
Undifferentiated connective tissue disease (UCTD), Uveitis, Vasculitis,
Vitiligo, Wegener's
granulomatosis (or Granulomatosis with Polyangiitis (GPA)), idiopathic
thrombocytopenia
purpura, splenomegaly;
(xxii) Systemic inflammatory response syndrome, cytokine release syndrome,
cytokine storm, immune reaction to a drug(s) or therapy(s), immune reaction to
an immune
activating drug(s) or agent(s) or treatment(s) or intervention(s), immune
reaction to
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CA 3050553 2019-07-25
immunotherapy and/or immune-oncology and/or immunomodulatory drug(s) and/or
treatment(s), adverse reaction to adoptive T-cell therapy(s), adverse reaction
to a chimeric
antigen receptor T-cell therapy(s) (CAR-T cell therapy(s)), adverse reaction
to a immune
checkpoint inhibitor(s), adverse reaction to monoclonal antibody drug(s),
tumor lysis
syndrome; or
(xxiii) cancer and Graft Versus Host Disease (GVHD) in transplantation therapy
in a
cancer patient,
wherein the method comprises administering to the subject an effective amount
of at
least one compound, or a composition containing at least one compound, which
reduces F1 Fo
ATP hydrolysis in a subject, or a pharmaceutically-acceptable salt, solvate,
hydrate or
prodrug thereof.
[35] A method according to Claim 16, wherein the subject is also administered
with an
effective amount of one or more compounds or compositions approved for human
use,
optionally for anti-cancer use, by the United States Food and Drug
Administration (FDA)
and/or European Medicines Agency (EMA), optionally in the same pharmaceutical
composition.
[36] A method according to Claim 17, wherein the subject is also administered
with an
effective amount of one or more compounds or compositions approved for human
use,
optionally for anti-cancer use, by the United States Food and Drug
Administration (FDA)
and/or European Medicines Agency (EMA), optionally in the same pharmaceutical
composition.
[37] A method according to Claim 16, wherein comparable or larger mg/kg
compound(s) ,
dose is used in, or administered to, a larger subject(s)/animal(s)/species,
which is very distinct
from most drugs, and optionally the mg/kg dosage administered to one or more
adult humans
is comparable or greater than the No Observed Adverse Effects Level (NOAEL)
mg/kg
dosage in mice housed at 22 C.
[38] A method according to Claim 17, wherein comparable or larger mg/kg
compound(s)
dose is used in, or administered to, a larger subject(s)/animal(s)/species,
which is very distinct
from most drugs, and optionally the mg/kg dosage administered to one or more
adult humans
is comparable or greater than the No Observed Adverse Effects Level (NOAEL)
mg/kg
dosage in mice housed at 22 C.
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[39] A method according to Claim 16, wherein the compound(s) and/or
composition(s) is
administered to the subject topically/locally and not systemically.
[40] A method according to Claim 17, wherein the compound(s) and/or
composition(s) is
administered to the subject topically/locally and not systemically.
[41] A compound(s) and/or composition(s) for use and/or method according to
any one of
Claims 1 to 33 or Claims 35 to 40, wherein the claim is made (e.g. written to
be/ammended
to be) a dependent claim of Claim 34.
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