Note: Descriptions are shown in the official language in which they were submitted.
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USE OF A MT KINASE INHIBITOR FOR TREATING OR PREVENTING BRAIN CANCER
FIELD OF THE INVENTION
The present invention is concerned with the finding that the macrocyclic
quinazoline
derivative 4,6-ethanediylidenepyrimido[4,5-b][6,1,12]benzoxadiazacyclo-
pentadecine,
17-bromo-8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl, described as
compound 22 in PCT publication W02004/105765, is useful in the manufacture of
a
medicament for the treatment of a primary brain cancer or treatment or
prevention of
brain metastasis. It accordingly provides methods for treating, preventing,
delaying or
mitigating brain metastasis and treating, delaying or mitigating primary brain
cancer.
BACKGROUND OF THE INVENTION
"Brain cancer" means (1) any abnormally increased proliferation of any type of
neuronal cell, hereinafter also referred to as primary brain cancer, or (2)
any other
cancer that has metastasized into the central nervous system (CNS),
hereinafter also
referred to as brain metastases.
Most neuronal cells- that is cells, that comprise or are found in the CNS,
including, for
example, neurons, microglia, and astrocytes- are "terminally differentiated",
meaning
that they no longer possess the ability to complete the cell cycle. (Kornblith
et al.,
(1986), Cancer: Principles and Practice of Oncology, 2nd Ed., DeVita, V.,
Hellman, S.,
Rosenberg, S, eds., J.B. Lippincott Company, Philadelphia, Chapter 41:
Neoplasms of
the Central Nervous System). Even if neuronal cells would enter the cell
cycle, they
are usually unable to complete the process as they would undergo apoptosis
(cell death)
(Multani, A.S., et al., Neoplasia 2(4), 339-45 (2000)). Only in those cases
where
neuronal cells lose the protective ability to undergo apoptosis, primary brain
cancers
may occur. Examples of primary brain cancers include, but are not limited to,
neuroma, astrocytoma, neuroblastoma, glioma, meningioma, oligodendroglioma,
medulloblastoma, spinal cord tumor and schwannoma.
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Gliomas comprise about 60% of all primary CNS tumors and usually occur in the
cerebral hemisphere of the brain, but may be found in other areas such as the
optic
nerve, brain stem or cerebellum. Gliomas are classified into groups according
to the
type of glial cell from which they originate (Komblith et al., (1986), Cancer:
Principles and Practice of Oncology, 2nd Ed., DeVita, V., Hellman, S.,
Rosenberg, S,
eds., J.B. Lippincott Company, Philadelphia, Chapter 41: Neoplasms of the
Central
Nervous System). The most common types of glioma are astrocytomas. These
tumors
develop from star-shaped glial cells called astrocytes. Astrocytomas are
assigned to
grades according to their malignancy. Low-grade astrocytomas, also known as
grade I
and II astrocytomas, are the least malignant, grow relatively slow and can
often be
completely removed using surgery. Mid-grade astrocytomas, also known as grade
III
astrocytomas, grow more rapidly and are more malignant. Grade III astrocytomas
are
treated with surgery followed by radiation and some chemotherapy. High-grade
astrocytomas, also known as grade IV astrocytomas, grow rapidly, invade nearby
tissue, and are very malignant. Grade IV astrocytomas are usually treated with
surgery
followed by a combination of radiation therapy and chemotherapy. Glioblastoma
multiforme are grade IV astrocytomas, which are among the most malignant and
deadly
primary brain tumors (Id). While the same surgical techniques and principles
have
applied to treating glioblastoma multiforme and less malignant brain tumors,
total
removal of a glioblastoma multiforme tumor has been more difficult to achieve.
The difference in malignancy is also reflected in the prognosis for a patient
having a
primary brain tumor. While a person treated for a grade I astrocytoma can
commonly
survive 10 years or more without recurrence, the mean length of survival for a
patient
with a grade IV astrocytoma tumor is 15 weeks after surgical treatment.
Because of the
high malignant-growth potential of grade IV astrocytoma tumors, only 5% of
patients
have survived for 1 year following surgical treatment alone, with a near 0%
survival
rate after 2 years. Radiation treatment in combination with surgical treatment
increases
the survival rate to about 10% after 2 years of treatment; however, virtually
no patients
survive longer than 5 years (Id).
While a treatment regimen of surgery, radiation therapy and chemotherapy
offers the
opportunity for a modestly increased lifespan for patients with a grade IV
astrocytoma
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brain tumor, the risks associated with each method of treatment are many. The
benefits
of treatment are minimal, and treatment can significantly decrease the quality
of the
patient's brief remaining lifespan. Accordingly, there remains a clear need in
the art for
primary brain cancer prevention and treatment methods that overcome the
disadvantages of the above-mentioned traditional approaches, in particular as
CNS
tumors represent the most common group of solid tumors in young patients.
Brain
tumors are the second-leading cause of cancer-related deaths in children,
accounting for
approximately 25% of all such deaths, and with the current therapies a
majority of these
children die within the first year of diagnosis.
Compared to primary brain tumors the incidence of brain metastases is much
higher.
Approximately 100,000 patients have symptomatic intracranial metastases in the
USA
annually and according to autopsy studies, a quarter of cancer patients have
tumor
metastases (Newton, H.B., et al., J. Neurooncol. 61, 35-44 (2003)). The brain
metastasis results from a primary tumor elsewhere in the body, include but are
not
limited to, for example, lung cancer (both small cell and non-small cell),
breast cancer,
colorectal cancer, prostate cancer, melanoma and pancreatic cancer. In
particular in
patients with metastatic breast cancer, the incidence of brain metastases is
diagnosed at
a rate of 10 to 20% (Tyson, R.M. et al., Therapy 3(1), 97-112 (2006)). Breast
cancer is
the second leading cause of cancer-related deaths in women and almost all
deaths from
breast cancer are due to metastatic disease with brain metastasis found in 30%
of
patients at autopsy.
The standard mode of treatment comprise surgical resection, chemotherapy and
radiation treatment in particular whole-brain radiotherapy (WBRT) and
stereotactic
radiosurgery (SRS) or a combination thereof. Surgery for brain metastasis can
improve
survival, especially in patients with single lesions. However, surgery may not
be
possible in the face of multiple lesions, surgically inaccessible lesions or
patients with
an inability to tolerate surgery. In breast cancer, 50% of the patients with
brain cancer
metastasis have multiple metastases, making them less suitable as surgical
candidates.
WBRT may improve median survival over no treatment, and as an adjuvant to
surgery
it reduces the recurrence rate and chances of dying a neurological death, but
it does not
change survival or level of function. SRS provides a method of treating brain
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metastasis that may be surgically unresectable, either by location or patient
condition.
SRS provides an improvement in quality of life but does not provide survival
benefit,
except in patients with a single metastasis. The use of chemotherapy in the
treatment
of brain metastasis is hampered by the inability of large molecular-weight
compounds,
thus restricting most chemotherapy agents, to cross the blood-brain barrier.
This is
reflected in the fact that chemosensitive tumors, such as most metastatic
breast
carcinomas, often show complete systemic responses to chemotherapy concomitant
with tumor progression in the brain. However, this initial reluctance to the
use of
chemotherapy in the treatment of brain metastasis did change over the recent
years
based on findings in animal models and human brain tumor autopsies, that
metastatic
lesions result in an impaired blood-brain barrier and by virtue of this,
chemotherapy
drugs can invariably enter into the tumor. Radiotherapy in conjunction with
chemotherapy has evolved into the first-line approach in the treatment of
brain
metastases.
However, there are serious limitations and dangers associated with all of the
current
methods of treatment. Radiotherapy, which often attempts to deliver highly
destructive
doses of ionizing radiation through the normal tissues of the body in an
attempt to
preferentially kill highly specific and often imperfectly defined areas of
cancerous
tissue, can have serious and significant side effects due to the destruction
of normal
nervous system or other tissues of the body, leading amongst others, to memory
loss
and personality alterations (infra). Chemotherapy, which attempts to
preferentially kill
cancerous cells instead of normal cells through the diverse administration of
chemical
agents or drugs to the tissues of the body, is limited in efficacy by the
chemical agents
currently available and can lead to toxic and unintended side effects on
normal tissue.
Surgery, which attempts to mechanically destroy or intervene in the
progression of
cancer, can also lead to serious side effects or consequences as a result of
mechanical
trauma or destruction of normal tissue. Some of the problems associated with
the above
approaches are (i) adverse side effects including alterations of intelligence,
learning
ability, memory, motor function, consciousness, and emotion; (ii) re-emergence
of the
tumor within three to five years of treatment due to development of resistance
to these
therapies; (iii) death due to the ineffectiveness of such treatment.
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From the foregoing, it will be apparent that there exists an urgently
compelling, yet
unsatisfied need to develop chemotherapeutic agents that can cross the blood-
brain
barrier in an amount effective to reduce growth and/or neoplastic spread of
the cancer
in the central nervous system.
SUMMARY OF THE INVENTION
The invention is directed in part to methods of treating or preventing brain
cancer,
and/or the treatment or prevention of brain metastasis utilizing certain
compounds
described in WO 2004/105765, the disclosure of which is hereby incorporated by
reference in its entirety.
In one embodiment, the present invention provides the use of the macrocyclic
quinazoline derivative 4,6-ethanediylidenepyrimido [4,5-b]
[6,1,12]benzoxadiazacyclo-
pentadecine, 17-bromo-8,9,10,11,12,13,14,19-octahydro-20-methoxy-13 -methyl,
described as compound 22 in PCT publication W02004/105765, in the manufacture
of
a medicament for the treatment or prevention of a primary brain cancer or
brain
metastasis.
In related embodiment, the invention provides a method of inhibiting
metastatic spread
of a cancer to the central nervous system, in a mammalian subject comprising
administering to a mammalian subject suspected of having metastatic cancer a
compound of the invention, in an amount effective to inhibit metastatic spread
of the
cancer to the central nervous system; and a method for treating brain cancer
comprising
administering to a mammalian subject diagnosed with a cancer a composition
comprising a compound of the invention, in an amount effect to reduce growth
or
neoplastic spread of the brain cancer/metastasis. It will be appreciated that
any
reduction in the rate of cancer growth or spread (which can prolong life and
quality of
life) is indicative of successful treatment. In preferred embodiments, cancer
growth is
halted completely. In still more preferred embodiments, cancers shrink or are
eradicated entirely. Preferred subjects for treatment are human subjects, but
other
animals, especially murine, rat, canine, bovine, porcine, primate, and other
model
systems for cancer treatment, are contemplated.
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In one variation of the foregoing methods of treatment, the compounds are
administered along with a second cancer therapeutic agent. The second agent
can be
any chemotherapeutic agent, radioactive agent, irradiation, nucleic acid
encoding a
cancer therapeutic agent, antibody, protein, and/or other anti-lymphangiogenic
agent or
an anti-angiogenic agent. The second agent may be administered before, after,
or
concurrently with the compounds of the invention.
In one variation, the subject to be treated has been diagnosed with an
operable tumor,
and the administering step is performed before, during, or after the tumor is
resected
from the subject. Compound treatment in conjunction with tumor resection is
intended
to reduce or eliminate regrowth of tumors from cancer cells that fail to be
resected.
Stated more generically, the invention provides a method of treating a brain
cancer, and
the treatment or prevention of brain metastasis comprising the step of
administering to
a mammal (including, but not limited to humans, rats, canines, bovines,
porcines, and
primates) in need thereof a compound of the invention.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1: Effect of 4,6-ethanediylidenepyrimido[4,5-
b] [6,1,12]benzoxadiazacyclopentadecine, 17-bromo-8,9,10,11,12,13,14,19-
octahydro-20-methoxy- 1 3-methyl (Compound 1) on s.c A431 tumor growth.
Gray bar indicates the treatment period with Compound 1, i.e. p.o., QDx14.
Black arrows indicate treatment with the reference compound BCNU
(Carmustin), i.e. i.v., Q14Dx2. Grey arrow - Day 18. On day 18 the median
subcutaneous (s.c.) tumor volumes were statistically analysed.
Figure 2: Statistical analysis of the median subcutaneous (s.c.) tumor volumes
of the
A431 cells. Gray bar indicates the treatment period with Compound 1, i.e.
p.o., QDx14.
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Figure 3: Kapalan Myer survival curves showing the survival of rats bearing
intracranial and subcutaneous tumors. On day 91 some surviving animals
start to be sacrificed due to extensive s.c tumor burden.
DETAILED DESCRIPTION OF THE INVENTION
WO-2004/105765 describes the preparation, formulation and pharmaceutical
properties
of macrocyclic quinazoline derivatives of formula (I) as multi targeted kinase
inhibitors
(MTKIs).
zl~ XZ 3' /, Ri
Y ~ 2' 1
J
z 6' R2
X~ 5 4
6/ ~
N 3 R3
Ra~ I /
7 ~ % 2 (1)
8 N1
It has now been found that one compound in the aforementioned class, i.e 4,6-
ethanediylidenepyrimido[4,5-b][6,1,12]benzoxadiazacyclo-pentadecine, 17-bromo-
8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl, described as compound 22
in
the aforementioned PCT publication, herein also referred to as MTKI 1 and/or
Compound 1, has clinical activity in brain cancer models and accordingly
provide the
use of this compound for the preparation of a pharmaceutical composition for
treating
brain cancer, including primary brain cancers and brain metastases as defined
hereinbefore.
Accordingly, in one aspect the present invention provides the use of 4,6-
ethanediylidenepyrimido[4,5-b] [6,1,12]benzoxadiazacyclo-pentadecine, 17-bromo-
8,9,10,11,12,13,14,19-octahydro-20-methoxy-13 -methyl or a pharmaceutically
acceptable acid or base addition salt thereof, in the manufacture of a
medicament for
the treatment or prevention of brain cancer.
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A further aspect of the present invention is directed to a method for the
treatment or
prevention of brain cancer in a mammalian subject, comprising administering a
therapeutically effective amount of 4,6-ethanediylidenepyrimido[4,5-
b] [6,1,12]benzoxadiazacyclo-pentadecine, 17-bromo-8,9,10,11,12,13,14,19-
octahydro-
20-methoxy- 13 -methyl or a pharmaceutically acceptable acid or base addition
salt
thereof, to a mammalian subject in need of such treatment.
The pharmaceutically acceptable acid or base addition salts as mentioned
hereinabove
are meant to comprise the therapeutically active non-toxic acid and non-toxic
base
addition salt forms which MTKI 1 is able to form. The basic properties can be
converted
in their pharmaceutically acceptable acid addition salts by treating said base
form with
an appropriate acid. Appropriate acids comprise, for example, inorganic acids
such as
hydrohalic acids, e.g. hydrochloric or hydrobromic acid; sulfuric; nitric;
phosphoric and
the like acids; or organic acids such as, for example, acetic, propanoic,
hydroxyacetic,
lactic, pyruvic, oxalic, malonic, succinic (i.e. butanedioic acid), maleic,
fumaric, malic,
tartaric, citric, methanesulfonic, ethanesulfonic, benzenesulfonic, p-
toluenesulfonic,
cyclamic, salicylic, p-aminosalicylic, pamoic and the like acids.
The acidic properties may be converted in their pharmaceutically acceptable
base
addition salts by treating said acid form with a suitable organic or inorganic
base.
Appropriate base salt forms comprise, for example, the ammonium salts, the
alkali and
earth alkaline metal salts, e.g. the lithium, sodium, potassium, magnesium,
calcium
salts and the like, salts with organic bases, e.g. the benzathine, N-methyl-D-
glucamine,
hydrabamine salts, and salts with amino acids such as, for example, arginine,
lysine and
the like.
The terms acid or base addition salt also comprise the hydrates and the
solvent addition
forms which MTKI 1 is able to form. Examples of such forms are e.g. hydrates,
alcoholates and the like.
In particular, the present invention is concerned with a use of the
dihydrobromide salt
of 4,6-ethanediylidenepyrimido[4,5-b][6,1,12]benzoxadiazacyclo-pentadecine, 17-
bromo-8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl, i.e., 17-bromo-
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8,9,10,11,12,13,14,19-octahydro-20-methoxy- 13 -methyl-4,6-
ethanediylidenepyrimido
[4,5-b][6,1,12]benzoxadiazacyclopentadecine dihydrobromide, in any of the
aforementioned uses for MTKI 1.
In a further embodiment, the present invention provides the use of the
aforementioned
MTKI 1 for the preparation of a pharmaceutical composition for the treatment
and/or
prevention of brain cancers.
The present invention also concerns a method of treating and/or preventing
brain
cancer in a mammal, comprising the step of administering a therapeutically
effective
amount of the aforementioned MTKI 1 to said mammal.
In a further embodiment, the present invention provides the use of the
aforementioned
MTKI 1 for the preparation of a pharmaceutical composition for the treatment
and/or
prevention of brain metastasis.
The present invention also concerns a method of treating and/or preventing
brain
metastasis in a mammal, comprising the step of administering a therapeutically
effective amount of the aforementioned MTKI 1 to said mammal.
Accordingly, in a further aspect, the most preferred compounds for use in
accordance
with the present invention are those selected from the group consisting of
compounds
having the following structure:
aBr
HN Br and HN 0 &a"
N
N O
O 0 N 2 HBr
The compounds according to the invention can be prepared and formulated into
pharmaceutical compositions by methods known in the art and in particular
according
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to the methods described in the published patent specification WO-2004/105765
mentioned herein and incorporated by reference.
A suitable preparation of the preferred compound used in this invention, taken
from
WO-2004/105765, follows:
Example 1
a) Preparation of 1-pentanol, 5-[[(4-bromo-2-nitrophenyl)methyl] amino]-
(intermediate
1)
A solution of 4-bromo-2-nitro- benzaldehyde,(0.013 mol), 5-amino-l-pentanol
(0.013 mol) and titanium, tetrakis (2-propanolato) (0.014 mol) in EtOH (15 ml)
was
stirred at RT for 1 hour, then the reaction mixture was heated to 50 C and
stirred for
30 min. The mixture was cooled to RT and NaBH4 (0.013 mol) was added
portionwise.
The reaction mixture was stirred overnight and then poured out into ice water
(50 ml).
The resulting mixture was stirred for 20 min., the formed precipitate was
filtered off
(giving Filtrate (I)), washed with H20 and stirred in DCM (to dissolve the
product and
to remove it from the Ti-salt). The mixture was filtered and then the filtrate
was dried
(MgSO4) and filtered, finally the solvent was evaporated. Filtrate (I) was
evaporated
until EtOH was removed and the aqueous concentrate was extracted 2 times with
DCM. The organic layer was separated, dried (MgSO4), filtered off and the
solvent was
evaporated, yielding 3.8 g (93 %) of intermediate 1.
Example 2
a) Preparation of 1-pentanol, 5-[[(4-bromo-2-nitrophenyl)methyl]methylamino]-
(intermediate 2)
A solution of intermediate 50 (0.0047 mol), formaldehyde (0.025 mol) and
titanium, tetrakis (2-propanolato) (0.0051 mol) in EtOH (150 ml) was heated to
50 C
and stirred for 1 hour, then NaBH4 (0.026 mol) was added portionwise at RT.
The
reaction mixture was stirred overnight and then quenched with water (100 ml).
The
resulting mixture was stirred for 1 hour; the formed precipitate was filtered
off and
washed. The organic filtrate was concentrated, then the aqueous concentrate
was
extracted with DCM and dried. The solvent was evaporated and the residue was
filtered
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over silica gel (eluent: DCM/CH3OH from 98/2 to 95/5). The product fractions
were
collected and the solvent was evaporated, yielding 0.5 g of intermediate 2.
b) Preparation of 1-pentanol, 5-[[(4-bromo-2-nitrophenyl)methyl]methylamino]-,
acetate (ester) (intermediate 3)
A solution of intermediate 2 (0.0015 mol) and pyridine (0.015 mol) in acetic
anhydride (8 ml) was stirred overnight at RT, then the solvent was evaporated
and co-
evaporated with toluene, yielding intermediate 3.
c) Preparation of 1-pentanol, 5-[[(2-amino-4-bromophenyl)methyl]methylamino]-,
acetate (ester) (intermediate 4)
A mixture of intermediate 3 (0.0015 mol) in THF (50 ml) was hydrogenated
with Pt/C 5% (0.5 g) as a catalyst in the presence of thiophene solution (0.5
ml) [H 179-
034]. After uptake of H2 (3 equiv.), the catalyst was filtered off and the
filtrate was
evaporated, yielding 0.5 g of intermediate 4.
d) Preparation of 6-quinazolinol, 4-[[2-[[[5-
(acetyloxy)pentyl]methylamino]methyl]-5-
bromophenyl]amino]-7-methoxy-, acetate (ester) (intermediate 5)
A mixture of intermediate 4 (0.0015 mol) and 4-chloro-7-methoxy-6-
quinazolinol acetate (ester) (0.0015 mol) in 2-propanol (30 ml) was heated to
80 C and
the reaction mixture was stirred for 1 day. The solvent was evaporated under
reduced
pressure and the residue was used as such in the next reaction step, yielding
0.83 g of
intermediate 5.
e) Preparation of 6-quinazolinol, 4-[[5-bromo-2-[[(5-
hydroxypentyl)methylamino]methyl]phenyl]amino]-7-methoxy- (intermediate 6)
A solution of intermediate 5 (0.0015 mol) in methanol (25 ml) was stirred at
RT
and a solution of K2C03 (0.003 mol) in H20 (2.5 ml) was added, then the
reaction
mixture was heated to 60 C and stirred for 18 hours. The solvent was
evaporated and
H20 (20 ml) was added, then the mixture was neutralized with acetic acid and
the
formed precipitate was filtered off. The filtrate was concentrated under
reduced
pressure and the concentrate was extracted with DCM, filtered, then dried
(MgSO4) and
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the mixture was concentrated under reduced pressure, yielding 0.5 g (70 %) of
intermediate 6.
Example 3
a)Preparation of 4,6-ethanediylidenepyrimido[4,5-b] [6,1,12]benzoxadiazacyclo-
pentadecine, 17-bromo-8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl-
(compound MTKI1)
A solution of intermediate 6 (0.0011 mol) in THF (50 ml) was stirred at RT and
tributylphosphine (0.0016 mol) was added, then 1,1'-(azodicarbonyl)bis-
piperidine
(0.0016 mol) was added and the reaction mixture was stirred for 2 hours. The
solvent
was evaporated until 1/3 of the initial volume. The resulting precipitate was
filtered off
and washed. The filtrate was evaporated and the residue was purified by RP
high-
performance liquid chromatography. The product fractions were collected and
the
organic solvent was evaporated. The aqueous concentrate was extracted 2 times
with
DCM and the organic layer was dried (MgSO4), then filtered off. The solvent
was
evaporated and the residue was dried (vac.) at 50 C, yielding 0.004 g (0.8 %)
of
compound MTKI1.
To prepare the aforementioned pharmaceutical compositions, a therapeutically
effective
amount of the particular compound, optionally in addition salt form, as the
active
ingredient is combined in intimate admixture with a pharmaceutically
acceptable
carrier, which may take a wide variety of forms depending on the form of
preparation
desired for administration. These pharmaceutical compositions are desirably in
unitary
dosage form suitable, preferably, for systemic administration such as oral,
percutaneous, or parenteral administration; or topical administration such as
via
inhalation, a nose spray, eye drops or via a cream, gel, shampoo or the like.
For
example, in preparing the compositions in oral dosage form, any of the usual
pharmaceutical media may be employed, such as, for example, water, glycols,
oils,
alcohols and the like in the case of oral liquid preparations such as
suspensions
(including nanosuspensions), syrups, elixirs and solutions; or solid carriers
such as
starches, sugars, kaolin, lubricants, binders, disintegrating agents and the
like in the
case of powders, pills, capsules and tablets. Because of their ease in
administration,
tablets and capsules represent the most advantageous oral dosage unit form, in
which
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case solid pharmaceutical carriers are obviously employed. For parenteral
compositions, the carrier will usually comprise sterile water, at least in
large part,
though other ingredients, for example, to aid solubility, may be included.
Injectable
solutions, for example, may be prepared in which the carrier comprises saline
solution,
glucose solution or a mixture of saline and glucose solution. Injectable
solutions
containing compounds of formula (I) may be formulated in an oil for prolonged
action.
Appropriate oils for this purpose are, for example, peanut oil, sesame oil,
cottonseed
oil, corn oil, soy bean oil, synthetic glycerol esters of long chain fatty
acids and
mixtures of these and other oils. Injectable suspensions may also be prepared
in which
case appropriate liquid carriers, suspending agents and the like may be
employed. In
the compositions suitable for percutaneous administration, the carrier
optionally
comprises a penetration enhancing agent and/or a suitable wettable agent,
optionally
combined with suitable additives of any nature in minor proportions, which
additives
do not cause any significant deleterious effects on the skin. Said additives
may
facilitate the administration to the skin and/or may be helpful for preparing
the desired
compositions. These compositions may be administered in various ways, e.g., as
a
transdermal patch, as a spot-on or as an ointment. As appropriate compositions
for
topical application there may be cited all compositions usually employed for
topically
administering drugs e.g. creams, gels, dressings, shampoos, tinctures, pastes,
ointments,
salves, powders and the like. Application of said compositions may be by
aerosol, e.g.
with a propellent such as nitrogen, carbon dioxide, a freon, or without a
propellent such
as a pump spray, drops, lotions, or a semisolid such as a thickened
composition which
can be applied by a swab. In particular, semisolid compositions such as
salves, creams,
gels, ointments and the like will conveniently be used.
It is especially advantageous to formulate the aforementioned pharmaceutical
compositions in dosage unit form for ease of administration and uniformity of
dosage.
Dosage unit form as used in the specification and claims herein refers to
physically
discrete units suitable as unitary dosages, each unit containing a
predetermined quantity
of active ingredient calculated to produce the desired therapeutic effect in
association
with the required pharmaceutical carrier. Examples of such dosage unit forms
are
tablets (including scored or coated tablets), capsules, pills, powder packets,
wafers,
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injectable solutions or suspensions, teaspoonfuls, tablespoonfuls and the
like, and
segregated multiples thereof.
Preferably, a therapeutically effective amount of the pharmaceutical
composition
comprising a compound according to the invention, is administered orally or
parenterally. Said therapeutically effective amount is the amount that
effectively
prevents metastasis and/or growth or reduces the size of a variety of
neoplastic disorders
or cell proliferative disorders (supra) in patients. On the basis of the
current data, it
appears that a pharmaceutical composition comprising a compound of the present
invention, and in particular
4,6-ethanediylidenepyrimido[4,5-b] [6,1,12]benzoxadiazacyclo-pentadecine, 17-
bromo-
8,9,10,11,12,13,14,19-octahydro-20-methoxy-13 -methyl (MTKI 1) as the active
ingredient can be administered orally in an amount of from 10 mg to several (1
to 5)
grams daily, either as a single dose or subdivided into more than one dose,
including,
e.g. two, three or even four times daily. A preferred amount ranges from 500
to 4,000
mg daily. A particularly, preferred dosage for such a compound is in the range
of 750
mg to 3,000 mg daily. It will be appreciated that the amount of a compound
according
to the present invention, also referred to here as the active ingredient,
which is required
to achieve a therapeutic effect will, of course, vary with, the route of
administration, the
age and condition of the recipient, and the particular disorder or disease
being treated.
The optimum dosage amounts and regimen can be readily determined by those
skilled in
the art using conventional methods and in view of the information set out
herein. This
treatment can be given either continuously or intermittently, including, e.g.
but not
limited to, cycles of 3-4 weeks with treatment given for 1-21 days per cycle
or other
schedules shown to be efficacious and safe.
One illustrative formulation is as follows:
Example 4: Formulation:
The product MTKI1 can be prepared as a 10-mg/mL oral solution, pH 2. It
contains an excipient, Captisol (chemical name: sulfobutyl ether-(3-
cyclodextrin,
SBE-(3-CD), citric acid, Tween 20, HCI, and NaOH in purified water. The
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formulation can be stored refrigerated (2-8 C; 36-46 F) and allowed to warm
to room
temperature for maximally 1 hour prior to dose preparation.
The product MTKI1 can also be prepared as 50-mg, 100-mg and 300-mg oral
immediate
release capsules, containing the active chemical entity MTKI1, lactose
monohydrate
(200 mesh), sodium lauryl sulphate and magnesium stearate in hard gelatin
capsules,
sizes 3, 4 and 00, respectively. The capsules may also contain any or all of
the following
ingredients: gelatin, red iron oxide and titanium oxide.
The above MTKI of the present invention may be used in combination with one or
more
other cancer treatments. Such combinations could encompass any established
antitumor
therapy, such as, but not limited to, chemotherapies, irradiation, and target
based therapies
such as antibodies and small molecules (including, but not limited to
Temozolomide or
BCNU). These therapies may be combined in systemic therapy, or local
instillation/administration (e.g. intrathecally), depending on optimum
efficacy/safety
requirements.
The MTKI 1 (e.g., Compound 1) and the further anti-cancer agent may be
administered
simultaneously (e.g. in separate or unitary compositions) or sequentially in
either order.
In the latter case, the two compounds will be administered within a period and
in an
amount and manner that is sufficient to ensure that an advantageous additive
or
synergistic effect is achieved. It will be appreciated that the preferred
method and order
of administration and the respective dosage amounts and regimens for each
component
of the combination will depend on the particular MTKI and further anti-cancer
agents
being administered, their route of administration, the particular tumor being
treated and
the particular host being treated. The optimum method and order of
administration and
the dosage amounts and regimen can be readily determined by those skilled in
the art
using conventional methods and in view of the information set out herein.
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EXPERIMENTAL DATA
The unique physico-chemical properties of MTKI I has resulted in an extremely
favourable tissue distribution profile including the ability to cross the
intact blood brain
barrier whilst still retaining good cellular activity and oral
bioavailability. Here, we
demonstrate that this preferential tissue distribution to the brain results in
significant
anti tumoral activity using experimental models of brain metastases.
A431 (ATCC, Rockville, MD. USA) vulval carcinoma cells were stereo-tactically
injected into athymic nude rats and mice. Tumor growth delay was followed by
using
animal survival as readout in the rat study or using MRI imaging in the case
of the
mouse study.
Our data demonstrate that MTKI I potently delays tumor growth, leading to
increased
survival in experimental models of brain metastases.
Methods
Brain models
Stereotaxic injection of cells in the brain of rats
Forty (40) rats (Wistar, BDIX or Sprague Dawley) in part I and 40 nude rats
were
sterotaxically injected with cells at DO. For stereotaxic injection of tumor
cells, rats
were anesthetised by an intramuscular injection of a Ketamine (Ketamine500 ,
Ref
043KET204, Centravet, France) and Xylazine (Rompun , Ref 002ROM001,
Centravet, France) mixture (2/1, v/v, 70 and 15 mg/kg, respectively). 1x105
A431 Cells
were stereotaxically injected using 4 independant stereotaxic apparatus (Kopf
Instrument, Germany and Stoelting Company, USA) in the right frontal lobe with
1X105 tumor cells re-suspended in 5 l of RPMI medium. Five l of the cell
suspension
was injected according to SOP No TEC-083/001 at 0.5 l/min. After cells
injection,
rats were observed during 1 hour.
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Subcutaneous injection of cells in rats
The same day of the stereotaxic injection, rats were injected subcutaneously
with A431
cells at DO. The tumor was obtained in rats by subcutaneous (SC) injection of
1x107
A431 cells in RPMI medium (200 l) in their right flank. For subcutaneous
injection of
tumor cells, rats were anesthetised by inhalation of Isoflurane Forene
(Minerve,
Bondoufle, France).
MTKI 1(Compound 1 in the figures hereinafter) was dissolved in a solution of
20%
Hydroxypropyl-(3-cyclodextrine (20% (3-HP-Cyclodextrin, pH 4.0) and given by
oral
gavage, daily for a period of fourteen days starting four days post
inoculation of the
cells, i.e. from day 4 - 17. Vehicle treated animals only received the (3-HP-
Cyclodextrin solution.
The reference compound BCNU (Carmustin) was dosed intravenously on day 4 and
day 18.
Results
Brain models
Subcutaneous injection of cells in rats
At the three doses tested, i.e. 50mpk, 75 mpk and 100 mpk, no clinical signs
of toxicity
such as body weight loss or behavioral changes were associated to the
treatment with
MTKI 1. Looking at the differences in median subcutaneous tumor volumes
observed
in the different animals (Figure 1), there was no effect of BCNU on A431 tumor
growth, whereas MTKI 1 clearly inhibited subcutaneous A431 tumor growth at all
doses tested. The high dose of BCNU (15 mpk) may have induced some mild
toxicity
(evident from sooner appearance of clinical signs of toxicity, i.e. body
weight loss) and
explains an even increased tumor growth rate when compared to the vehicle
controls.
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On day 18, i.e. 24 hr after the last MTKI 1 treatment cycle, statistical
analysis of the
s.c. A431 tumors (Figure 2) showed a significant effect of MTKI 1 on s.c.
tumor
growth at all doses tested. Treatment versus Control (taking control as 100%)
values
were 5.37% for 50 mpk of MTKI 1, 4.29% for 75 mpk of MTKI 1 and 3.17% for 100
mpk of MTKI 1.
Stereotaxic injection of cells in the brain of rats
The Kapalan Myer survival curves (Figure 3) clearly show the survival benefit
for
animals treated with MTKI 1(Compound 1). A dose-dependent marked effect of
MTKI 1(Compound 1) on animal survival time (few `accidental' animal death were
observed following documented mis-administration of the compound in the lungs,
for
instance, and/or not related to the presence of brain tumors) was observed.
Survival
times (50%) changed from 49 for vehicle treated animals to 61, 83 and 88 days
for
animals treated with 50 mpk, 75 mpk and 100 mpk of Compound 1 respectively.
In these experiments, some animals surviving at day 91 had to be sacrificed
due to the
extensive s.c. tumor burden.
While the foregoing specification teaches the principles of the present
invention, with
examples provided for the purpose of illustration, it will be understood that
the practice
of the invention encompasses all of the usual variations, adaptations and/or
modifications as come within the scope of the following claims and their
equivalents.
