Note: Descriptions are shown in the official language in which they were submitted.
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HIV PROTEASE INHIBITORS
BACKGROUND AND SUMMAR~i' OF THE INVENTION
This invention relates to a novel series of chemical
compounds useful as HI:V protease inhibitors and to the use
of such compounds as antiviral agents.
Acquired Immune L>eficiency Syndrome (AIDS) is a rela-
tively newly recognized disease or condition. AIDS causes a
gradual breakdown of the body's immune system as well as
progressive deterioration of the central and peripheral
nervous systems. Since its initial recognition in the early
1980's, AIDS has spread rapidly and has now reached epidemic
proportions within a relatively limited segment of the
population. Intensive research has led to the discovery of
the responsible agent, human T-lymphotropic retrovirus III
(HTLV-III), now more commonly referred to as the human
immunodeficiency virus; or HIV.
HIV is a member c>f the class of viruses known as
retroviruses. The ret.roviral genome is composed of RNA
which is converted to DNA by reverse transcription. This
retroviral DNA is then stably integrated into a host cell's
chromosome and, emplo~~ing the replicative processes of the
host cells, produces new retroviral particles and advances
the infection to other cells. HIV appears to have a
particular affinity for the human T-4 lymphocyte cell which
plays a vital role in the body's immune system. HIV
infection of these white blood cells depletes this white
cell population. Eventually, the immune system is rendered
inoperative and ineffective against various opportunistic
diseases such as, among others, pneumocystic carini
pneumonia, Kaposi's sarcoma, and cancer of the lymph system.
Although th.e exact mechanism of the formation and work-
ing of the HIV virus i.s not understood, identification of
the virus has le:d to some progress in controlling the
disease. For example, the drug azidothymidine (AZT) has
been found effective f:or inhibiting the reverse
transcription of the retroviral genome of the HIV virus,
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thus giving a measure of control, though not a cure, for
patients afflicted with AIDS. The search continues for
drugs that can cure or at least provide an improved measure
of control of the deadly HIV virus.
Retroviral replication routinely features post-
translational processing of polyproteins. This processing
is accomplished by virally encoded HIV protease enzyme.
This yields mature polypeptides that will subsequently aid
in the formation and function of infectious virus. If this
molecular processing is stifled, then the normal production
of HIV is terminated. Therefore, inhibitors of HIV protease
may function as anti-HIV viral agents.
HIV protease is one of the translated products from the
HIV structural protein pol gene. This retroviral protease
specifically cleaves other structural polypeptides at
discrete sites to release these newly activated structural
proteins and enzymes, thereby rendering the virion
replication-competent. As such, inhibition of the HIV
protease by potent compounds may prevent proviral
integration of infected T-lymphocytes during the early phase
of the HIV-1 life cycle, as well as inhibit viral
proteolytic processing during its late stage. Additionally,
the protease inhibitors may have the advantages of being
more readily available, longer lived in virus, and less
toxic than currently available drugs, possibly due to their
specificity for the retroviral protease.
In accordance with this invention, there is provided a
novel class of chemical compounds that can inhibit and/or
block the activity of the HIV protease, which halts the
proliferation of HIV virus, pharmaceutical compositions
containing these compounds, and the use of the compounds as
inhibitors of the HIV protease.
The present invention relates to compounds falling
within formula (9) below, and pharmaceutically acceptable
salts, prodrugs, and solvates thereof, that inhibit the
protease encoded by human immunodeficiency virus (HIV) type
1 (HIV-1) or type 2 (HIV-2). These compounds are useful in
the treatment of infection by HIV and the treatment of the-
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acquired immune deficiency syndrome (AIDS). The compounds,
their pharmaceutically acceptable salts, and the
pharmaceutical compositions of the present invention can be
used alone or in combination with other antivirals,
immunomodulators, antibiotics or vaccines. Compounds of the
present invention ca.n also be used as prodrugs. Methods of
treating AIDS, methods of treating HIV infection and methods
of inhibiting HIV protease are disclosed.
The compounds of the present invention are of the
formula (9):
R
Me O SPh O~N=R'
HO , _
H OH N H
H
9
wherein:
R and R' <~re independently selected from H, a
substituted or unsubstituted alkyl-OR1 group, a cycloalkyl
group substitui~ed with a (C,-C6) alkyl group or a (C1-
C~)alkyl-OH group, a heterocycle group substituted with a
(C1-C6) alkyl gr~~up or a (C,-C6) alkyl-OH group, an alkyl-NRzR3
group, or an a:Lkyl-S (X) (Y) R4 group,
wherein
R1 is H, a. substituted or unsubstituted alkyl group, or
an acyl group;
RZ and R3 ;ire each independently selected from H,
substituted or mnsubstituted alkyl, cycloalkyl,
heterocyc_Le, and aryl groups, and acyl and sulfonyl
groups;
Rq is H, a substituted or unsubstituted alkyl,
cycloalky~_, het~=_rocycle, or aryl group; and
X and Y are each independently selected from =O and
nothing;
or a pharmaceut:icall~~ acceptable prodrug, salt or solvate
thereof.
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Preferably in the compounds of formula 9, R is H. More
preferably, R is H and R' is a cycloalkyl group selected
f rom
CHzOH CH3 CHzOH CHzOH
. and
Preferably in the compounds of formula 9 when at least one
of R and R' is an alkyl-OR1 group, R, is H. Particularly
when at least one of R and R' is an alkyl-OR1 group, the
alkyl-ORl is selected from -C (CH3) ZCHZOH, -CH (CH3) CHZOH,
-CHzCHzOH, -C (CH3) (CHzOH) 2, -C (CH3) ?-O-CHI-O-CH3, -C (CHI) zCH2-O-
CHZ-O-CH3, and -C (CH3) ZCH2-O-acyl, or a pharmaceutically
acceptable prodrug, salt or solvate thereof.
Preferably when at least one of R and R' is a
cycloalkyl group substituted with a (C,-C6)alkyl group or a
(C1-C6)alkyl-OH group, the cycloalkyl group is selected from:
s CH3 ' CH3 _ CH2QH CH3 CHzOH CHzOH
. and
Preferably when at least one of R and R' is a heterocycle
group substituted with a (C1-C6) alkyl group or a (C1-
C6)alkyl-OH group, the heterocycle group is selected from:
H3C H3C HaC C H3
'S
O° S' , S ,
H3C CH3 H3C R3 CH3 H3C
N /
O ° ~ , C IN-R3> and
wherein R3 is H, a substituted or unsubstituted alkyl,
cycloalkyl, heterocycle, or aryl group, or an acyl or
sulfonyl group.
A preferred species of the formula (9) is [3S-
[2(2S*,3S*),3 alpha,4a beta,8a beta]]-N-(1,1-dimethyl-2-
hydroxyethyl)decahydro-2-[2-hydroxy-3-[(3-hydroxy-2-
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methylbenzoyl)~~mino]-4-(phenylthio)butyl]-3-
isoquinolineca:rboxamide
H
Me O SPh O~N
~~ OH
H~~ ' Me Me
y ~N wN
H ~ H
H
21
and its pharmaceutically acceptable salts, and its prodrug
analogs. Preferred prodrugs can be obtained by replacing
the hydrogen in one of the alcohol groups with an acyl
group, and more preff~rably an amino acid acyl group.
The present invention further provides pharmaceutical
formulations compris_Lng an effective amount of a compound of
formula (9) or a pharmaceutically acceptable salt thereof,
in combination with a pharmaceutically acceptable carrier,
such as a dilue~nt or excipient.
The present invention further provides a method of
treating AIDS compri:~ing administering to a host or patient,
such as a primate, an effective amount of a compound of the
present invention.
The present invention further provides a method of
inhibiting HIV replication comprising administering to an
HIV infected cell, a cell susceptible to HIV infection or a
host or patient, such as a primate, an effective amount of a
compound of the present invention.
Det;~iled Description of the Invention
The present invE:ntion provides new compounds falling
within formula (9), as described above, that are useful for
treating HIV infection and/or AIDS.
Applicants incorporate by reference U.S. Patent No.
5,484,926, U.S. Patent Application Nos. 08/708,411 and
08/708,607, and Japanese Patent Application Nos. JP 95-
2481.83 and JP 95-2481.84, with the caveat that the
definitions of preferences, terms, variables, labels and the
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like used in each application are applicable only to the
corresponding disclosure from that application.
In particular, since each of the above-identified
applications incorporated by reference was prepared
separately, the original applications may use in some
instances the same term, label or variable to mean something
different. For example, the variable "X" is used in each
application, but each application has its own distinct
definition of the substituent or moiety represented by this
variable. It will be apparent to those skilled in the art
that the terms, labels and variables in each application
incorporated by reference are limited solely to the
disclosure from that application, and may be replaced by
other suitable terms, labels and variables or the like
representing the particular substituents and moieties. Of
course, those skilled in the art will realize that any
suitable set of terms, labels and variables may be used to
generically or more specifically represent the subject
matter disclosed in the present application, including
terms, labels, variables, and the like universally
applicable to the incorporated disclosures of the
above-identified applications and the following disclosure.
Compounds of the formula (9) may be prodrugs, which can
serve to improve the pharmaceutical properties of the
compounds, such as pharmacokinetic properties, for example,
improved bioavailability or solubility. The preparation of
prodrugs may be carried out by standard methods known to
those skilled in the art. A preferred prodrug can be
obtained by acylation or alkylation of the starting alcohol
when R or R' is CH ( CH3 ) ZCHZOH .
All temperatures stated herein are in degrees Celsius
(°C). All units of measurement employed herein are in
weight units except for liquids which are in volume units.
The term "alkyl" as used herein refers to straight or
branched chain groups, preferably, having one to eight, more
preferably having one to six, and most preferably having
from one to four carbon atoms. The term "C1-C6 alkyl"
represents a straight or branched alkyl chain having from.
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one to six carbon atoms. Exemplary C1-C6 alkyl groups
include methyl,, ethyl, n- propyl, isopropyl, butyl,
isobutyl, sec-butyl, t-butyl, pentyl, neo-pentyl, hexyl,
isohexyl, and t:he like. The term "C1-C6 alkyl" includes
within its def_Lnition the term "C1-CQ alkyl" .
The term "cyclo<~lky1" represents a saturated or
partially saturated, mono- or poly-carbocylic ring,
preferably having 5-:14 ring carbon atoms. Exemplary
cycloalkyls include monocyclic rings having from 3-7,
preferably 3-6, carbon atoms, such as cyclopropyl,
cyclobutyl, cyc:lopent:yl, cyclohexyl, cycloheptyl and the
like. An exemplary cycloalkyl is a C,-C~ cycloalkyl, which
is a saturated hydrocarbon ring structure containing from
five to seven carbon atoms.
The term "alkoxyl" represents -O-alkyl. An example of
an alkoxyl is a C1-C6 alkoxyl, which represents a straight or
branched alkyl chain having from one to six carbon atoms
attached to an oxygen atom. Exemplary C1-C~ alkoxyl groups
include methox:yl, et:hoxyl, propoxyl, isopropoxyl, butoxyl,
sec-butoxyl, t-butoxyl, pentoxyl, hexoxyl, and the like.
C1-C6 alkoxyl include; within its definition a C1-Cq alkoxyl.
The term "aryl" as used herein refers to a carbocyclic
or heterocyclic, aromatic, 5-14 membered monocyclic or
polycyclic ring. Exemplary aryls include phenyl, naphthyl,
anthryl, phenan.thryl, thienyl, pyrrolyl, imidazolyl,
pyrazolyl, furyl, isothiazolyl, furazanyl, isoxazolyl,
thiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl,
triazinyl, benzo[b]thienyl, naphtho[2,3-b]thianthrenyl,
isobenzofuranyl, chromenyl, xanthenyl, phenoxathienyl,
indolizinyl, isoindolyl, indolyl, indazolyl, purinyl,
isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl,
quinoxyalinyl, quinzolinyl, benzothiazolyl, benzimidazolyl,
tetrahydroquinolinyl, cinnolinyl, pteridinyl, carbazolyl,~
beta-carbolinyl, phenanthridinyl, acridinyl, perimidinyl,
phenanthrolinyl, phen.azinyl, isothiazolyl, phenothiazinyl,
and phenoxazinyl.
The term "arylox.yl" represents -O-aryl.
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_g_
The term "hydrolyzable group" is a group, which when
bonded to an oxygen, forms an ester, which can be hydrolyzed
in vivo to a hydroxyl group. Exemplary hydrolyzable groups,
which are optionally substituted, include acyl function,
sulfonate function and phosphate function. For example,
such hydrolyzable groups include blocked or unblocked amino
acid residue, a hemisuccinate residue, and a nicotinate
residue.
The term "halogen" represents chlorine, fluorine,
bromine or iodine. The term "halo" represents chloro,
fluoro, bromo or iodo.
The term "carbocycle" represents an aromatic or a
saturated or a partially saturated 5-14 membered monocyclic
or polycyclic ring, such as a S- to 7-membered monocyclic or
7- to 10-membered bicyclic ring, wherein all the ring
members are carbon atoms.
The term "heterocycle" represents an aromatic or a
saturated or a partially saturated, 5-14 membered, monocylic
or polycyclic ring, such as a 5- to 7-membered monocyclic or
7- to 10-membered bicyclic ring, having from one to three
heteroatoms selected from nitrogen, oxygen and sulfur, and
wherein any nitrogen and sulfur heteroatoms may optionally
be oxidized, and any nitrogen heteroatom may optionally be
quaternized. The heterocyclic ring may be attached at any
suitable heteroatom or carbon atom. Examples of such
heterocycles include decahydroisoquinolinyl,
octahydro-thieno[3,2-c]pyridinyl, piperidinyl, piperazinyl,
azepinyl, pyrrolyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl,
imidazolyl, isobenzofuranyl, furazanyl, imidazolinyl,
imidazolidinyl, pyridyl, pyrazinyl, pyrimidinyl,
pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl,
thianthrenyl, triazinyl, isoxazolidinyl, morpholinyl,
thiazolyl, thiazolidinyl, isothiazolyl, quinuclidinyl, iso-
thiazolidinyl, indolyl, quinolinyl, chromenyl, xanthenyl,
isoquinolinyl, benzimidazolyl, thiadiazolyl, benzopyranyl,
benzothiazolyl, benzoazolyl, furyl, tetrahydrofuryl,
tetrahydropyranyl, thienyl, benzothienyl, benzo[b]thienyl,
naphtho[2,3-b]thienyl, thiamorpholinyl,
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thiamorpholiny:Lsulfoxide, thiamorpholinylsulfone,
oxadiazolyl, t:riazolyl, tetrahydroquinolinyl, tetrahydriso-
quinolinyl, phc~noxathienyl, indolizinyl, isoindolyl,
indazolyl, pur:inyl, isoquinolyl, quinolyl, phthalazinyl,
naphthyridinyl,, quinoxyalinyl, quinzolinyl,
tetrahydroquinolinyl, cinnolinyl, pteridinyl, carbazolyl,
beta-carboliny=L, phenanthridinyl, acridinyl, perimidinyl,
phenanthroliny=L, phenazinyl, isothiazolyl, phenothiazinyl,
and phenoxazinyl.
The term "thioether" includes S-aryl, such as
phenylthio and naphtlzylthio; S-heterocycle where the
heterocycle is saturated or partially saturated;
S- (CS-C~) -cycloalkyl; and S-alkyl, such as C,-C~ alkylthio.
In the thioether, the=_ -aryl, the -heterocycle, the
-cycloalkyl anc~ the --alkyl can optionally be substituted.
An example of a thioether is "C1-C6 alkylthio", which
represents a st;raight~ or branched alkyl chain having from
one to six carbon atoms attached to a sulfur atom.
Exemplary Cl-C6 alkylthio groups include methylthio,
ethylthio, propylthio, isopropylthio, butylthio, sec-
butylthio, t-bL~tylth_Lo, pentylthio, hexylthio, and the like.
The term "mercapto" represents -SH.
The term "amino" represents -NL1L2, wherein Ll and LZ
are preferably indepE:ndently selected from oxygen,
carbocycle, het.erocyc:le, alkyl, sulfonyl and hydrogen; or
NC (0) L3, wherein L3 is preferably alkyl, alkoxyl, hydrogen or
-NL~L~. The ar~~l, alltyl and alkoxyl groups can optionally be
substituted. A,n example of an amino is C1-C9 alkylamino,
which represents a straight or branched alkyl chain having
from one to four carbon atoms attached to an amino group.
Exemplary C1-C9 alkylamino groups include methylamino,
ethylamino, propylam.i_no, isopropylamino, butylamino, sec-
butylamino, and. the 7.ike. Another example of an amino is
di(C1-C9)alkylamino, which represents two straight or
branched alkyl chain~~, each having from one to four carbon
atoms attached to a common amino group. Exemplary di(C1-
CQ)alkylamino groups include dimethylamino,
ethylmethylamino, met:hylpropylamino, ethylisopropylamino,
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butylmethylamino, sec-butylethylamino, and the like. An
example of an amino is C1-Cq alkylsulfonylamino, which has a
straight or branched alkyl chain having from one to four
carbon atoms attached to a sulfonylamino moiety. Exemplary
C1-Cq alkylsulfonylamino groups include methylsulfonylamino,
ethylsulfonylamino, propylsulfonylamino,
isopropylsulfonylamino, butylsulfonylamino, sec-
butylsulfonylamino, t-butylsulfonylamino, and the like.
The term "acyl" represents L6C(O)L4, wherein L6 is a
single bond, -O or -N, and further wherein Lq is preferably
alkyl, amino, hydroxyl, alkoxyl or hydrogen. The alkyl and
alkoxyl groups can optionally be substituted. An exemplary
acyl is a C1-C~ alkoxycarbonyl, which is a straight or
branched alkoxyl chain having from one to four carbon atoms
attached to a carbonyl moiety. Exemplary C,-CQ
alkoxycarbonyl groups include methoxycarbonyl,
ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl,
butoxycarbonyl, and the like. Another exemplary acyl is a
carboxy wherein L6 is a single bond and Lq is alkoxyl,
hydrogen, or hydroxyl. A further exemplary acyl is N-(C1-
C9) alkylcarbamoyl (L6 is a single bond and L4 is an amino) ,
which is a straight or branched alkyl chain having from one
to four carbon atoms attached to the nitrogen atom of a
carbamoyl moiety. Exemplary N-(C1-Cq)alkylcarbamoyl groups
include N-methylcarbamoyl, N-ethylcarbamoyl,
N-propylcarbamoyl, N-isopropylcarbamoyl, N-butylcarbamoyl,
and N-t-butylcarbamoyl, and the like. Yet another exemplary
acyl is N,N-di(C1-C4)alkylcarbamoyl, which has two straight
or branched alkyl chains, each having from one to four
carbon atoms attached to the nitrogen atom of a carbamoyl
moiety. Exemplary N,N-di(C1-C4)alkylcarbamoyl groups include
N,N-dimethylcarbamoyl, N,N-ethylmethylcarbamoyl, N,N-
methylpropylcarbamoyl, N,N-ethylisopropylcarbamoyl, N,N-
butylmethylcarbamoyl, N,N-sec-butylethylcarbamoyl, and the
like.
The term "sulfinyl" represents -SO-L5, wherein LS is
preferably alkyl, amino, aryl, cycloalkyl or heterocycle.
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The alkyl, aryl, cycloalkyl and heterocycle can all
optionally be substituted.
The term "sulfonyl" represents -SOz-L5, wherein LS is
preferably alkyl, aryl, cycloalkyl, heterocycle or amino.
The alkyl, aryl, cycloalkyl and heterocycle can all
optionally be substituted. An example of a sulfonyl is a
C1-Cq alkylsulfonyl, which is a straight or branched alkyl
chain having from one to four carbon atoms attached to a
sulfonyl moiety. Exemplary C1-Cq alkylsulfonyl groups
include methylsulfonyl, ethylsulfonyl, propylsulfonyl,
isopropylsulfo:zyl, butylsulfonyl, sec-butylsulfonyl, t-
butylsulfonyl ;end the like.
As indical~ed above, many of the groups are optionally
substituted. :Ln fact, unless specifically noted, all of the
groups defined by the terms defined in this application may
be substituted or unsubstituted. For instance, when the
term "alkyl" i:~ used, it should be understood to encompass
both substituted and unsubstituted alkyl unless specific
exclusion of one or the other is positively stated.
Examples of substituents for alkyl and aryl include
mercapto, thioether, nitro (NOZ), amino, aryloxyl, halogen,
hydroxyl, alko~tyl, and acyl, as well as aryl, cycloalkyl and
saturated and partially saturated heterocycles. Examples of
substituents for hetE~rocycle and cycloalkyl include those
listed above for alkyl and aryl, as well as aryl and alkyl.
Exemplary substituted aryls include a phenyl or
naphthyl ring :~ubstii:uted with one or more substituents,
preferably one to three substituents, independently selected
from halo, hydx-oxy, rnorpholino (C1-CQ) alkoxy carbonyl, pyridyl
(C1-C~) alkoxycarbonyl, halo (C1-Cq) alkyl, C1-C9 alkyl, C1-C9
alkoxy, carbox~~, C1-C'q alkoxycarbonyl, carbamoyl,
N- (Ci-C9) alkylcarbamo:yl, amino, C1-Cq alkylamino,
di (C1-CQ) alkyl amino o:r a group of the formula
-(CH2)a-R~ where a is 1, 2, 3 or 4; and ~.~ is hydroxy, C1-C~
alkoxy, carboxy, C1-Cq alkoxycarbonyl., amino, carbamoyl,
C1-C9 alkyl amino or d:i (C1-C9) alkyl amino.
Another substituted alkyl is halo(C,-Cq)alkyl, which
represents a straight: or branched alkyl chain having from-
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one to four carbon atoms with 1-3 halogen atoms attached to
it. Exemplary halo(C1-C9)alkyl groups include chloromethyl,
2- bromoethyl, 1-chloroisopropyl, 3-fluoropropyl,
2,3-dibromobutyl, 3-chloroisobutyl, iodo-t-butyl,
trifluoromethyl and the like.
Another substituted alkyl is hydroxy(C1-CQ)alkyl, which
represents a straight or branched alkyl chain having from
one to four carbon atoms with a hydroxy group attached to
it. Exemplary hydroxy(C1-CQ)alkyl groups include
hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxy-
isopropyl, 4-hydroxybutyl and the like.
Yet another substituted alkyl is C1-C9 alkylthio(C;-
C4) alkyl, which is a straight or branched Cl-C9 alkyl group
with a C1-CG alkylthio group attached to it. Exemplary C1-C,
alkylthio(C1-Cq)alkyl groups include methylthiomethyl,
ethylthiomethyl, propylthiopropyl, sec-butylthiomethyl, and
the like.
Yet another exemplary substituted alkyl is
heterocycle(C1- C9)alkyl, which is a straight or branched
alkyl chain having from one to four carbon atoms with a
heterocycle attached to it. Exemplary
heterocycle(C1-CQ)alkyls include pyrrolylmethyl, quinolinyl-
methyl, 1-indolylethyl, 2-furylethyl, 3-thien-2-ylpropyl, 1-
imidazolylisopropyl, 4-thiazolylbutyl and the like.
Yet another substituted alkyl is aryl(C1-Cq)alkyl, which
is a straight or branched alkyl chain having from one to
four carbon atoms with an aryl group attached to it.
Exemplary aryl(Cl-C9)alkyl groups include phenylmethyl,
2-phenylethyl, 3-naphthyl-propyl, 1-naphthylisopropyl,
4-phenylbutyl and the like.
The heterocycle can, for example, be substituted with
1, 2 or 3 substituents independently selected from halo,
halo (C~- CQ ) alkyl , C,-Cq alkyl , C1-C4 alkoxy, carboxy, CI-C9
alkoxycarbonyl, carbamoyl, N-(C,-CQ)alkylcarbamoyl, amino,
C1-CQalkylamino, di (C1-CQ) alkyl amino or a group having the
structure - (CHZ) a-R' where a is 1, 2, 3 or 4 and R' is
hydroxy, C2-CQ alkoxy, carboxy, C1-Cq alkoxycarbonyl, amino,
carbamoyl, C1-Cq alkyl amino or di (C,-C4) alkylamino.
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Examples of substituted heterocycles include
3-N-t-butyl carboxam.ide decahydroisoquinolinyl, 6-N-t-butyl
carboxamide octahydro-thieno[3,2-c]pyridinyl,
3-methylimidazolyl, 3-methoxypyridyl, 4-chloroquinolinyl,
4-aminothiazolyl, 8-methylquinolinyl, 6-chloroquinoxalinyl,
3-ethylpyridyl, 6-methoxybenzimidazolyl, 4-hydroxyfuryl,
4-methylisoqui:aolinyl, 6,8-dibromoquinolinyl, 2-methyl-
1,2,3,4-tetrahydroisoquinolinyl, N-methyl-quinolin-2-yl,
2-t-butoxycarbc~nyl-1,2,3,4-isoquinolin-7-yl and the like.
Exemplary heterocyclic ring systems represented by A or
B include (1) !~-membered monocyclic ring groups such as
thienyl, pyrro:lyl, ivmidazolyl, pyrazolyl, furyl,
isothiazolyl, :Euraza:nyl, isoxazolyl, thiazolyl and the like;
(2) 6-membered monocyclic groups such as pyridyl, pyrazinyl,
pyrimidinyl, p~~ridazinly, triazinyl and the like; and (3)
polycyclic hete~rocyc:lic rings groups, such as
decahydroisoquinolinyl, octahydro-thieno [3,2-c] pyridinyl,
benzo [b] thieny:L , naphtho [2 , 3 -b] thianthrenyl ,
isobenzofurany:L,_chromenyl, xanthenyl, and fully or
partially saturated analogs thereof.
A cycloallcyl ma~~ be optionally substituted with 1, 2 or
3 substituents indepE=_ndently selected from halo,
halo (C~-C~) alky:L, C1-C9 alkyl, C1-Cq alkoxy, carboxy, C1-C4
alkoxycarbonyl , carbamoyl , N- ( C, -CQ ) alkyl carbamoyl , amino ,
Cl-Cq alkyl amino, di (Ci-C~) alkylamino or a group having the
structure - (CH2) a-R' where a is 1, 2, 3 or 4 and R' is
hydroxy, Cl-C9 alkoxy, carboxy, C1-Cq alkoxycarbonyl, amino,
carbamoyl, Cz-C9 alkylamino or di (C,-C9) alkyl amino.
Exemplary substituted cycloalkyl groups include
3-methylcyclopE:ntyl, 4-ethoxycyclohexyl,
5-carboxycyclo-~hepty:L, 6-chlorocyclohexyl and the like.
Exemplary substituted hydrolyzable groups include
N-benzyl glycyl., N-Cbz-L-valyl, and N-methyl nicotinate.
The compounds of the present invention have at least
five asymmetric: centers denoted by an asterisk in the
formula (9) below:
SUBSTITUTE SHEET (RULE 26)
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SPh
Me O Oa-N:R
HO N** N
H OH H
H
9
As a consequence of these asymmetric centers, the
compounds of the present invention can occur in any of the
possible stereoisomeric forms, and can be used in mixtures
of stereoisomers, which can be optically active or racemic,
or can be used alone as essentially pure stereisomers, i.e.,
at least 95% pure. All asymmetric forms, individual
stereoisomers and combinations thereof, are within the scope
of the present invention.
The individual stereoisomers may be prepared from their
respective precursors by the procedures described above, by
resolving the racemic mixtures, or by separating the
diastereomers. The resolution can be carried out in the
presence of a resolving agent, by chromatography or by
repeated crystallization or by some combination of these
techniques which are known in the art. Further details
regarding resolutions can be found in Jacques et al.,
Enantiomers, Racemates, and Resolutions, John Wiley & Sons
1981.
Preferably, the compounds of the present invention are
substantially pure, i.e, over 50o pure. More preferably,
the compounds are at least 75% pure. Even more preferably,
the compounds are more than 90% pure. Even more preferably,
the compounds are at least 95% pure, more preferably, at
least 97o pure, and most preferably at least 99o pure.
As mentioned above, the invention includes the
pharmaceutically acceptable salts of the compounds defined
by formula (9). A compound of this invention may possess a
sufficiently acidic, a sufficiently basic, or both
functional groups, and accordingly react with any of a
number of inorganic or organic bases, and inorganic and
organic acids, to form a pharmaceutically acceptable salt.
SUBSTITUTE SHEET (RULE 26)
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The term "pharmaceutically acceptable salt", as used
herein, refers to salts of the compounds of the above
formula which ;ire substantially non-toxic to living
organisms. Exemplary pharmaceutically acceptable salts
include those ;alts prepared by reaction of the compounds of
the present im;rention with a mineral or organic acid or an
inorganic base. The reactants are generally combined in a
mutual solvent such .as diethylether or benzene, for acid
addition salts,, or water or alcohols for base addition
salts. The sa_Lts normally precipitate out of solution
within about one hour to about ten days and can be isolated
by filtration or other conventional methods. Such salts are
known as acid addition and base addition salts.
Acids that: may be employed to form acid addition salts
are inorganic ciCldS such as hydrochloric acid, hydrobromic
acid, hydroiod~_c acid, sulfuric acid, phosphoric acid, and
the like, and organic acids such as p-toluenesulfonic,
methanesulfonic: acid,, oxalic acid, p-bromophenylsulfonic
acid, carbonic acid, succinic acid, citric acid, benzoic
acid, acetic acid, and the like.
Examples of pharmaceutically acceptable salts are the
sulfate, pyrosulfate, bisulfate, sulfite, bisulfate,
phosphate, monohydroc~enphosphate, dihydrogenphosphate,
metaphosphate, pyrophosphate, chloride, bromide, iodide,
acetate, propionate, decanoate, caprylate, acrylate,
formate, isobutyrate, caproate, heptanoate, propiolate,
oxalate, malona.te, succinate, suberate, sebacate, fumarate,
maleate, butyne-1,4-c~ioate, hexyne-1,6-dioate, benzoate,
chlorobenzoate, methylbenzoate, dinitrobenzoate,
hydroxybenzoate, methoxybenzoate, phthalate, sulfonate,
xylenesulfonate, phenylacetate, phenylpropionate,
phenylbutyrate, citrate, lactate, g-hydroxybutyrate,
glycollate, tartrate, methane-sulfonate, propanesulfonate,
naphthalene-1-sulfonate, napththalene-2-sulfonate, mandelate
and the like.
Preferred pharmaceutically acceptable acid addition
salts are those formed with mineral acids such as
hydrochloric acid anal hydrobromic acid, and those formed
SUBSTITUTE SHEET (RULE 26)
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with organic acids such as malefic acid and methanesulfonic
acid.
Base addition salts include those derived from
inorganic and organic bases, such as ammonium or alkali or
alkaline earth metal hydroxides, carbonates, bicarbonates,
and the like. Such bases useful in preparing the salts of
this invention thus include sodium hydroxide, potassium
hydroxide, ammonium hydroxide, potassium carbonate, sodium
carbonate, sodium bicarbonate, potassium bicarbonate,
calcium hydroxide, calcium carbonate and the like. The
potassium and sodium salt forms are particularly preferred.
A "pharmaceutically acceptable prodrug" is intended to
mean a compound that may be converted under physiological
conditions or by solvolysis to a compound of the formula 9.
A "pharmaceutically acceptable solvate" is intended to
mean a solvate that retains the biological effectiveness and
properties of the biologically active components of
compounds of formula 9.
Examples of pharmaceutically acceptable solvates
include, but are not limited to, compounds of formula 9 in
combing ion with water, isopropanol~, ethanol, methanol,
DMSO, ethyl acetate, acetic acid, or ethanolamine.
It should be recognized that the particular counterion
forming a part of any salt of this invention is not of a
critical nature, so long as the salt as a whole is
pharmacologically acceptable and as long as the counterion
does not contribute undesired qualities to the salt as a
whole.
SUBSTITUTE SHEET (RULE 26)
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A preferred compound is compound 21
H
Me O sPh O~N
~~ OH
HO - Me Me
1 \ ~N N
lI H OH H
H
21
[3S- [2 (2S*, 3S* ) , 3 alpha, 4a beta, 8a beta] ] -N- (1, 1-dimethyl-2-
hydroxyethyl)decahydro-2-[2-hydroxy-3-[(3-hydroxy-2-
methylbenzoyl)amino)-4-(phenylthio)butyl]-3-
isoquinolinecarboxamide.
A process for making compound 21 is provided below.
Compound 21 ha:~ also been obtained as a metabolite from the
plasma of patients administered [3S-
(3R,4aR*,8aR*,a?'S*,3'S*)]-2-[2'-hydroxy-3'-phenylthiomethyl-
4' -aza-5' -oxo-Vii' - (2 " -methyl-3 "-hydroxyphenyl) pentyl]
decahydroisoquinolinc=_-3-N-t-butylcarboxamide methanesulfonic
acid salt, which is disclosed in U.S. Patent No. 5,484,926.
The compounds o:E formula 9 can be prepared according to
the following F:eaction Scheme I.
SUBSTITUTE SHEET (RULE 26)
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REACTION SCHEME I
Scheme I. General Synthetic Pathway for the Production of 96 and Derivatives
R
CONHt-Bu OOOH COOH R R~ Oy N' R'
aqums
H' N H Mdto~s H' N H Ami~ePrdedon ~~ N . H ' H Rp~ N
--w ----s H
H Step la H Step Ib H
Artidecwpiog H
Step 1
to 2a
2b Rp =artcx prteauggoup 3
SPh
R
CbzHN~CI R R
O~N R SPh O tV SPh O~N,R,
H.N OH 5 ~_ 'R~ Chrrertnval
H CHotoarohol qty N ~ H_N~N
L~proteaon ~ OFI H OH ~H
H epoxide Step xI~lIS
Step 3 cbse-open seqmrce H H
q 6
Step 4
Me
AcO~COQ R R
Me p SPh O~ N, R, Me O SPh O~ N, R
acetate
g Ac0 ~ ~' mmval
NFI Y 'N HO ~ ~
H NH Y 'N
antis coplo~ OH ~ OH H
Step 6a H~ ~eP 66
9a H
9b
Compound 1a, perhydroisoquinoline, which is
commercially available from NSC Technologies (Chicago, IL)
or Procos SpA (Milan, Italy) is subjected to prolonged acid
hydrolysis in step 1a to obtain compound 2a. A variety of
inorganic acids may be used in either an aqueous/organic
solvent mixture or in water alone at temperatures above 50
°C. An example of such an inorganic acid is 6N aqueous HCl.
Substitutes for compound la include the corresponding esters
lb, thioesters lc or other amides ld:
H COOZ H COSZ CONZ~ZZ
H
H H
H
H ~ ~ H
lb lc ld ,
where Z, Z1 and Zz may each independently be alkyl,
cycloalkyl, heterocycle, or aryl.
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Compound 2~~ is then protected at the amine nitrogen to
obtain compound 2b in step Ib. The protecting group Rp is
defined as a suitably conjugating group to avoid unwanted
decomposition oi= activated carboxylate derivatives of
compound 2b in ~>tep 2. Such protecting groups typically can
be carbamate in origin, having a general structure of
formula 11:
O
R"O~
11
The identity of R" in formula 11 can be any alkyl,
cycloalkyl, aryl, or heterocycle which can be removed easily
in a deprotection step after Step,2. Examples of R"
include, but are not limited to methyl, ethyl, propyl,
isopropyl, n-butyl, i:~obutyl, t-butyl or higher branched or
unbranched alkyl, 2,2,2-trichloroethyl, 2-
trimethylsilylethyl, allyl, phenyl, substituted phenyl,
benzyl, substituted benzyl, 9-fluorenylmethyl, 9-
anthrylmethyl anal higher polycyclic aromatic ring system.
The following materials, as defined below, can be obtained
from the Aldrich Chemical Co. (Sigma Aldrich Fluka):
a
2.2,2-trichloroethyl= -a-12-C-a
a
H a~3
2-trimethylsllylethyl= -~2-C-SI--a'13
H a-13
allyl= -a-IZ-CH=Cii2
henzyl= _ q.~ / \
9-flourenylmethyl =
I
9-anihrylnethyt =
SUBSTITUTE SHEET (RULE 26)
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Such protecting groups typically can be installed by an
acylation reaction of the corresponding haloformate ester
12a or a dicarbonate 12b:
O O
I~'O~X R"O~Y
12a 12b
X = halogen Y = OCOR"
in the presence of a suitable base in typical organic
solvents for these types of reactions such as halogenated
solvents, ethers and hydrocarbons. Such bases are typically
inorganic, such as metal hydroxides, bicarbonates and
carbonates or organic bases such as amines like
triethylamine, diethylamine, diethyl isopropylamine, 1,8-
diazabicyclo[2.2.2)octane (DABCO) or related di- or
trialkyl-amines, as well as amidine bases like 1,8-
diazabicyclo [5.4 . 0) under-7-ene (DBU) and 1, 8-
diazabicyclo[4.3.0]non-5-ene (DBN). The following
materials, as defined below, can be obtained from the
Aldrich Chemical Co. (Sigma Aldrich Fluka):
~aaoo =
oBU = ~~
These reactions are typically run anywhere from below
room temperature to approximately 100 °C.
The amide coupling Step 2 can be accomplished in any
number of fashions depending on how the carboxyl group is
activated. A group J is installed in Step 2 by reaction of
the carboxylic acid 2b to produce the activated derivative
2c.
SUBSTITUTE SHEET {RULE 26)
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Sten 2
J R
COOH 05! I
activaW nofthe Rpm R~N~R O~ N~R
N H carboxyl goup N H H ~~ N
-----~ H
H H ~ base
H
2b Rp = am~e protect~g goy 2c J = leav~g goup 3
The group ~T can be any of a variety of leaving groups
such as alkoxy, hydroxy, halogen, pseudohalogen (including
azide, cyanide, isocy<~nate and isothiocyanate), alkyl or
arenesulfonate, aromatic heterocycle(bonded through a
heteroatom) and N-hyd:roxyheterocycle, including
hydroxysuccinimide or hydroxybenzotriazole ester. The
following definitions apply to the terms above:
azide -N-t~N
cyanide -C=N
isocyanate-N=C=O
isothiocyanate-t~~g
O
alkylsulfonate-O-S-alkyl
i
O
O
ii
arenesuNonate-O-S-aryl
O
N-hydroxyhe:erocyclic HO-
where N~ = nitrogen heterocycle
\~/O\\
N-hydroxysuccinimide HO-N
O
hyd roxybe nnotriazo le
~I N
N
OH
SUBSTITUTE SHEET (RULE 26)
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The aryl halides (2c, J = halogen) may be prepared
using inorganic halogenating agents such as thionyl chloride
or bromide, phosphorous trichloride or bromide, phosphorous
pentachloride or bromide or organic agents such as oxalyl
chloride or trichlorisocyanuric acid. Esters (2c, J = OR")
(R" is defined above) may be prepared in a variety of ways
starting from the acid chloride 2c where J is Cl by
combination with the desired alcohol in the presence of an
organic or inorganic base stated previously for the
acylation of compound 12a or compound 12b. Alternatively,
the ester may be produced by acid-promoted esterification in
the presence of the desired alcohol. The sulfonates (2c, J =
OSO~Wl, where Wl is alkyl or aryl) are typically made by
reaction of the carboxylic acid 2b with alkyl or
arylsulfonyl chlorides in the presence of an organic amine
base such as triethylamine in a non-polar solvent at
temperatures below 0 °C. Alkyl and arylsulfonyl are defined
as follows:
0
alkylx~lfonylchloride = d-S-alkyl
O
O
arenesulfonyl chloride = d-S-aryl
O
The pseudohalogen derivatives of 2c (J = pseudohalogen)
are typically made from the acid halides 2c (J = halogen) by
reaction with inorganic pseudohalide in the presence of a
base. Such bases include, but are not limited to metal
hydroxides, bicarbonates and carbonates or organic bases
such as amines like triethylamine, diethylamine, diethyl
isopropylamine, 1,8-diazabicyclo[2.2.2]octane (DABCO) or
related di- or trialkylamines, as well as amidine bases like
1,8-diazabicyclo[5.4.0]under-7-ene (DBU) and 1,8-
diazabicyclo[4.3.0]non-5-ene (DBN). A particularly
preferred base is triethylamine. The heteroaromatic
SUBSTITUTE SHEET (RULE 26)
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derivatives of 2c are also made from the acid halides 2c (J
- halogen), utilizing the specific heteroaromatic compound
in the presence of an amine base in a non-polar solvent.
The N-hydroxyher_erocyclic derivatives of 2c can be made from
the acid halide; as above and may also be generated using
alkyl carbodiim:ides (.alkyl-N=C=N-alkyl, where the alkyl
groups can be the same or different) or aryl carbodiimides
(aryl-N=C=N-ary:L, where the aryl groups can be the same or
different) and an amine base as condensing agents.
The primar~r or secondary amine (shown above the arrow
in Step 2 of Scheme I,? used in the coupling process may
incorporate suitable protecting groups, depending on the
functionality present in the amine and the mode of coupling
used. The mode of coupling of 2c with a primary or
secondary amine can bE=_ carried out in a variety of ways
depending on the' ident=ity of J. 4~Then a free acid is used
(2c, J = OH) the. coupling can be performed using
carbodiimide-ba~~ed methods utilizing any of the common
reagents of thi:~ clas:~, including dicyclohexylcarbodiimide
or related dialk:ylcarbodiimides, EDC (salts of 1- (3-
dimethylaminopropyl)-.3-ethylcarbodiimide) or related water-
soluble reagent; along with an organic amine base in polar
organic solvent; such as dioxane, DMF, NMP and acetonitrile
in the presence of an N-hydroxyheterocyclic compound such as
N-hydroxysuccini.mide or 3-hydroxybenzotriazole.
Alternatively, halofox°mate esters, such as 12d, may be used
to temporarily a.ctivat:e the acid to give mixed anhydrides of
general formula 2d.
O~OR"
O:yO
R~~N- O
H
R"O~ X
H'
12d X = halogen
2d
SUBSTITUTE SHEET (RULE 26)
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Such haloformate esters are typically as shown in 12d above
and include methyl-, ethyl-, isopropyl-, isobutyl-, n-butyl,
phenyl- and related alkyl and aryl chloroformates, defined
below.
0
alkyl chloiofortnate = 'I
alkyl-O~Q
O
aryl chlorofortnate = aryl-O~C!
Formula 2d is a possible intermediate in the step from
formula 2b to formula 3. Formula 2d is an intermediate, but
the process described here results in formula 3, without
isolation of Formula 2d.
These reactions are typically performed in a variety of
non-polar organic solvents like halocarbons and ethers such
as diethyl ether, methyl t-butylether, diisopropyl ether,
dioxane and THF at temperatures below 0 °C accompanied by an
organic amine base such as triethylamine, diethylamine,
diethyl isopropylamine, DABCO or related di- or
trialkylamines, as well as amidine bases like DBU and DBN.
When J in compound 2c is an alkyl or arenesulfonate (J
- OSOZR or OSO2Ar), the coupling can be performed in a
variety of non-polar organic solvents like halocarbons and
ethers, such as diethyl ether, methyl t-butylether,
diisopropyl ether, dioxane and THF at temperatures below 0
°C, accompanied by an organic amine base such as
triethylamine, diethylamine, diethyl isopropylamine, DABCO
or related di- or trialkylamines, as well as amidine bases
like DBU and DBN.
When J in compound 2c is a halogen or pseudohalogen,
the coupling may be performed in most common organic
solvents such as THF, diethyl ether, dioxane, methyl t-butyl
ether or other ethers; acetone, cyclohexanone, methyl
isobutylketone and other ketones; esters such as ethyl,
SUBSTITUTE SHEET (RULE 26)
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methyl and isopropyl acetate; halogenated solvents such as
halogenated metlzanes and ethanes, chlorobenzene and other
halogenated ben:~enes; nitrites such acetonitrile and
propionitrile; :Lower alcohols such as ethanol, isopropanol,
t-butanol and related alcohols; and polar organic solvents
such as dimethy:Lformamide, dimethylsulfoxide, N-methyl-2-
pyrrollidone and related amide-containing solvents. A base
is frequently used and may be any of a number of inorganic
bases such as metal hydroxides, bicarbonates and carbonates
or organic baser such as amines like triethylamine,
diethylamine, d~_ethyl isopropylamine, DABCO or related di-
or trialkylamines, as well as amidine bases like DBU and
DBN.
One skillecL in the art will be able to perform the
amide coupling ~~tep 2 with other possible J groups.
In Step 3 protect=ing group removal can be accomplished
using any of the standard methods for deprotecting a
particular clas:~ of protecting group. Simple alkyl- and
substituted alk~.Tl~carbamates can be removed with aqueous
solutions of ba:>e at t=emperatures up to about 100 °C,
employing any of: the common inorganic metal hydroxides such
as sodium-, litriium-, potassium- or barium hydroxide or
hydroxides of other metals in at least stoichiometric
amounts. Carbamate protecting groups that contain benzyl
groups bonded tc> oxygen may be removed by hydrogenolysis
with a palladium or platinum catalyst. Alternatively,
aqueous base hycLrolysis may be used at temperatures up to
about 100 °C, employing any of the common inorganic metal
hydroxides such as sodium-, lithium-, potassium- or barium
hydroxide or hydroxides of other metals in at least
stoichiometric a.mount~~. A variety of anhydrous acids may
also be used for deprotection of benzyl-based carbamates,
including HCl, H:Br and HI. Lewis acids of boron and
aluminum such as A1C13, BBr3, BClj in non-polar solvents are
also effective. Certain substituted benzyl, aryl or alkyl
groups in which the specific substitution pattern is chosen
for its ability to be removed under specific conditions may
also be used. F'or example, the 2-
SUBSTITUTE SHEET {RULE 26)
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trimethylsilylethylcarbonyl group (Teoc) is a protecting
group designed to take advantage of the specific reactivity
of the 2-trimethylsilylethyl group in the deprotection
process. 2-Trimethylsilylethylcarbonyl chloride may be used
to protect the amine nitrogen and may later be removed using
sources of fluoride ion such as HF or tetraalkylammonium
fluoride salts.
In Step 4, the perhydroisoquinoline piece of formula 4
is connected to the Chloroalcohol (compound 5, Scheme I) via
an epoxide intermediate (13) generated via the base-induced
closure of the vicinal chlorohydrin functionality.
SPh
CbzHN
O
13
Compound 5 is produced by Kaneka Industries, Japan. Several
close-open procedures in proceeding from compound 5
compound 13 ~ compound 6 may be used. The epoxide 13 may be
isolated or it may be reacted with 4 added either subsequent
to formation of 13 or 4 may be present from the beginning of
the sequence. The epoxide 13 can be generated using
inorganic bases such as metal hydroxides, carbonates and
bicarbonates in solvents such as alcohols like methanol
ethanol or isopropyl alcohol, ethers such as THF and dioxane
or mixtures of the two. The epoxide can also be generated in
a 2-phase solvent system consisting of water and a
halocarbon solvent such as dichloromethane along with the
base. A phase-transfer catalyst such as a
tetraalkylammonium salt may be used to facilitate the
process. The process of opening the epoxide 13 with
compound 4 is accomplished in alcohol solvents or mixtures
of an alcohol and another solvent which may be an ether or a
Bipolar aprotic solvent such as dimethylformamide or
dimethylsulfoxide. The opening of the epoxide 13 with
SUBSTITUTE SHEET (RULE 26)
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compound 4 to give compound 6 is optimally performed over a
period of 2-7 hours at. 50 - 60 °C.
In Step 5 t:he car_bobenzyloxy group can be removed to
give the free amine 7.. This can be done using HBr in acetic
acid using cosol.vents such as halocarbons. It can also be
performed using halide's of boron such as BBrj and BC13 or
alkyl substituted boron halides such as dimethylboron
bromide in halocarbon solvents like chloroform and
dichloromethane at temperatures ranging from 0 °C up to
ambient temperature. Alternatively, the carbobenzyloxy
group can be removed by hydrolysis using aqueous/alcoholic
solutions of metal hydroxides like barium, sodium, lithium
or potassium hydroxide at temperatures above ambient for
periods of hours.
Step 6a is the coupling of benzoic acid derivatives of
formula 8 to give 9a. In Formula 8, Q can be a leaving
group. Q can be any of: the leaving groups discussed above
for Group J. The compounds of formula 8 where Q = OH or Cl
are commercially available from EMS Dottikon, Lenzburg,
Switzerland and Sugai Chemical Industries, Ltd. in Japan.
The coupling can be carried out in a variety of ways,
depending on the identity of Q. When a free acid is used (Q
- OH), the coupling ca.n be performed using carbodiimide
based methods utilizing any of the common reagents of this
class including dicyclohexylcarbodiimide or related
dialkylcarbodiimides, EDC (salts of 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide) or related water
soluble reagents along with an organic amine base in polar
organic solvents such as dioxane, DMF, NMP and acetonitrile
in the presence of an N-hydroxyheterocyclic including N-
hydroxysuccinimide or 3-hydroxybenzotriazole. When Q = a
halogen or pseudohalogen, the coupling may be performed in
most common organic solvents such as THF, diethyl ether,
dioxane, methyl t-butyl ether or other ethers; acetone,
cyclohexanone, methyl isobutylketone and other ketones;
esters such as ethyl, methyl and isopropyl acetate;
SUBSTITUTE SHEET (RULE 26)
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halogenated solvents such as halogenated urethanes and
ethanes, chlorobenzene and other halogenated benzenes;
nitrites such acetonitrile and propionitrile; lower alcohols
such as ethanol, isopropanol, t-butanol and related
alcohols, and polar organic solvents such as
dimethylformamide, dimethylsulfoxide, N-methyl-2-
pyrrollidone and related amide-containing solvents. A base
is frequently used and may be any of a number of inorganic
bases such as metal hydroxides, bicarbonates and carbonates
or organic bases such as amines like triethylamine,
diethylamine, diethyl isopropylamine, DABCO or related di-
or trialkylamines, as well as amidine bases like DBU and
DBN.
Acetate removal is accomplished in step 6b with aqueous
or alcoholic solutions of inorganic bases such as metal
hydroxides, carbonates and bicarbonates at ambient
temperatures up to 100 °C. If there is a protected
functionality on the carboxamide group bonded to the
perhydroisoquinoline ring system, it is best removed at this
point (during or after step 6b). The, nature of this step is
dependent on the exact identity of the protecting group.
A preferred method for accomplishing the entire process
shown in Scheme I is shown in Scheme II.
The Cbz-protected amino acid 15 was coupled with the amine
22 to give the amide 16. The Cbz group was removed by
hydrogenation to
SUBSTITUTE SHEET (RULE 26)
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Scheme 11. Synthesis of Amidc 21
H ~ H
I
H COM-I-t-Bu COOIH ~l7 fMS
OII
N 1. 6 N aq. HC1 ~ N H=N ~ ~~ H2 H Me
H -~. H ~ N 5% Pd~C 1 N H
H
2. CI~I H FDC, f-Ipgt~H20 H EtOH
H
Pattptltnisoquinoune STEP A STEP 8 -
STEP C "
sPh
Me
C» CI SPh p~ 1V H ~ I \ COC1
OH ~ _ ()I1 SPh N
Me 50°/ aq. thpt-1 ~ pl I ~ 20
CLlotvalaohd Cbal W N H
OH HzN N
IJaCXI, IPA H IPA ~ ~i fi Et3N, THF, EtOH;
Ij ~ then 50%aq. N~OII
STEP D '8 STEP E '9 STEP F
H
SPh
Me Oti
H OH N . H
H
21
give the amine :L7. This was coupled with the chloroalcohol
via the epoxide using the in situ procedure to give the
adduct 18. Com~entional deprotection with base and coupling
of the free primary amine with the acid chloride 20 gave
rise to amide 2:1. Details of this process are provided
below in ExamplE~s 1 A to F. The lettering A to F in Scheme
II corresponds i.o Examples 1 A to F below.
The following Examples illustrate aspects of the
invention. There examples are for illustrative purposes and
are not intended to limit the scope of the invention.
Abbreviations for the terms melting point, nuclear
magnetic resonance spectra, electron impact mass spectra,
field desorption mass spectra, fast atom bombardment mass
spectra, infrared spectra, ultraviolet spectra, elemental
analysis, high performance liquid chromatography, and thin
layer chromatography are, respectively, m.p., NMR, EIMS,
MS(FD), MS(FAB), IR, W, Analysis, HPLC, and TLC. In
addition, the absorption maxima listed for the IR spectra
are those of ini=erest, not all maxima observed.
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In conjunction with the NMR spectra, the following
abbreviations are used: singlet (s), doublet (d), doublet
of doublets (dd) , triplet (t) , quartet (q) , multiplet (m) ,
doublet of multiplets (dm), broad singlet (br.s), broad
doublet (br.d), broad triplet (br.t), and broad multiplet
(br.m). J indicates the coupling constant in Hertz (Hz).
Unless otherwise noted, NMR data refer to the free base of
the subject compound.
NMR spectra were obtained on a General Electric QE-300
300MHz instrument. Chemical shifts are expressed in b
values in ppm. Mass spectra were obtained on a VG ZAB-3
Spectrometer at the Scripps Research Institute, La Jolla,
CA. Infra-red spectra were recorded on a Midac Corporation
spectrometer. UV spectra were obtained on a Varian,Cary 3E
instrument. Thin layer chromatography was carried out using
silica plates available from E. Merck. Melting points were
measured on a Mettler FP62 instrument and are uncorrected.
Example 1
Procedures for the Synthesis of Amide of Formula 21
[3S- [2 (2S*, 3S*) , 3 alpha, 4a beta, 8a beta] ] -N- (l, 1-dimethyl-2-
hydroxyethyl)decahydro-2-[2-hydroxy-3-[(3-hydroxy-2-
methylbenzoyl)amino]-4-(phenylthio)butyl]-3-
isoquinolinecarboxamide
H
SPh O N
Me O ~ ~OH
HO ~ Me Me
N
H OH
i
21
A. Perhydroisoquinoline (26.4 g, 111 mmol} (commerically
available from NSC Technologies (Chicago, IL) or Procos SpA
(Milan, Italy)) was suspended in water (200 mL) and
concentrated aqueous HC1 (200 mL). This mixture was heated
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to reflux and stirred for 3 days, during which time it went
into solution. The solvents were removed under reduced
pressure to give a light yellow solid. The solid was
slurried in 2-propanol (200 mL) and filtered. The filtrate
was evaporated under reduced pressure to an oil. EtOAc (100
mL) and water (100 mL) were added and the pH of the solution
was brought to 8.0 by the addition of 2 N aqueous KOH.
Benzyl chloroformate (15.8 mL, 111 mmol) was added dropwise
over 30 minutes and the pH was kept between 7 and 8 by the
addition of 2 N aqueous KOH. The mixture was stirred at
room temperature for 18 hours. EtOAc (200 mL) was added and
the organic layer was washed with 1 N aqueous HC1 (100 mL),
and brine (100 mL). The organic layer was dried (MgS09),
filtered, and evaporated under reduced pressure to an oil.
The product was purified by silica gel chromatography,
eluting with 1:1 40-60 petroleum ether/EtOAc followed by
1000 EtOAc. The fractions containing product were collected
and evaporated under reduced pressure to give the compound
15 (11.3 g, 32%) as a colorless oil: 1H NMR (300 MHz, CDC13)
~ 7.43-7.28 (m, 5 H), 5.17 (br s, 2 H), 4.76 (m, 1 H), 3.79
(m, 1 H), 3.33 (m, 1 H), 2.19 (m, 1 H), 1.96 (m, 1 H), 1.88-
1.15 (m, 10 H) .
B. 1-Hydroxyb~~nzotriazole (4.2 g, 31.4 mmol) and EDC (6.0
g, 31.4 mmol) ware added to a solution of acid 15 (8.3 g,
26.2 mmol) in DI~IF (128 mL) at ambient temperature. The
mixture was hea":,ed at 80° C for 10 minutes. 1,1-Dimethyl-2-
trimethylsilylo:Kyethylamine (5.1 g, 31.4 mmol, prepared from
1,1-dimethyl-2-lzydroxyethylamine (Aldrich Chemical Co.) and
hexamethyldisil;~zane (Aldrich Chemical Co.)) by heating the
mixture neat under reflux for several hours followed by
evaporation of the volatile components was added and the
solution was he~~ted at 80° C for 17 hours. The yellow
solution was po~ired into EtOAc (250 mL) and 2 N aqueous HCl
(250 mL). Afte:r stirring for 10 minutes EtOAc (750 mL) was
added and the mixture was washed with H20 (3 x 500 mL) and
brine (1 x 250 mL). The combined aqueous layers were
extracted with 7~tOAc (1 x 250 mL). The combined organic
layers were dried (NazS04) and purified by flash
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chromatography (50/50 EtOAc/hexanes) to give the compound 16
as a colorless oil (7.9 g, 78%): 1H NMR (300 MHz, CD30D) S
7.36 (m, 5 H), 5.20 (d, J = 8.1 Hz, 1 H), 5.10 (m, 1 H),
4.53 (m, 1 H), 3.78 (dd, J = 13.2, 4.4 Hz, 1 H), 3.60 (m, 2
H), 3.48 (d, J = 10.7 Hz, 1 H), 2.15-1.25 (m, 12 H), 1.31
(s, 3 H) , 1.29 (s, 3 H) .
C. A mixture of carbamate 16 (7.9 g, 20.4 mmol) and 5%
palladium on carbon (Pd/C)(1.6 g) was hydrogenated at 50 psi
H~ in absolute EtOH (110 mL) at ambient temperature for 18
hours. The mixture was filtered through Celite and
evaporated in vacuo to give amine 17 as a white, crystalline
solid: 'H NMR (300 MHz, CD30D) b 3.63 (q, J = 7.0 Hz, 2 H),
3.34 (m, 1 H), 3.27 (dd, J = 11.8, 3.3 Hz, 1 H), 2.91 (m, 1
H), 2.02-1.15 (m, 12 H), 1.32 (s, 3 H), 1.31 (s, 3 H).
D. Aqueous 10.2 N NaOH (2.4 mL, 24.5 mmol) was added to a
warm (27 °C) suspension of chloroalcohol (obtained from
Kaneka Industries in Japan)(10.4 g, 28.6 mmol) in
isopropanol (IPA)(104 mL) with mechanical stirring. After 1
hour 1 N aqueous HC1 in IPA (prepared by addition of 1 mL of
concentrated aqueous HCl to 12 mL of IPA) approximately
(ca.) 1 mL) was added to neutralize (pH = 7). Amine 17 (5.2
g, 20.4 mmol) was added as a solution in IPA (50 mL) and the
thin suspension was heated at 60° C for 10 hours. The IPA
was removed in vacuo. The residue was diluted with EtOAc
(150 mL) and washed with Hz0 (2 x 50 mL) , saturated aqueous
NaHC03 (1 x 50 mL), and brine (1 x 50 mL). The combined
aqueous layers were extracted with EtOAc (1 x 25 mL). The
combined organic layers were dried (NazSO~) and purified by
flash chromatography (75/25 EtOAc/hexanes, then EtOAc) to
give the compound 18 as a white solid (8.98 g, 76%): 1H NMR
(300 MHz, CD30D) d 7.33 (m, 10 H) , 5.08 (AB, JAB = 12.2 Hz,
~uAB = 12.1 Hz, 2 H), 3.96, (m, 2 H), 3.56 (q, J = 7.3 Hz, 2
H), 3.50, (m, 1 H), 3.20 (dd, J = 13.6, 9.2 Hz, 1 H), 3.03
(m, 1 H) , 2.64 (m, 2 H) , 2.20-1.20 (m, 14 H) , 1.28 (s, 6 H) .
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E. 50% aqueous NaOH (2.7 g, 1.8 mL, 33.6 mmol) was added
to a suspension of carbamate 18 (6.75 g, 11.6 mmol) in IPA
(34 mL) at ambient temperature. The mixture was heated
under reflux for 12 hours. After cooling to ambient
temperature, the mixture was diluted with methyl t-butyl
ether (MTBE) (600 mL) and washed with H20 (2 x 250 mL) and
brine (1 x 125 mL). T'he combined aqueous layers were
extracted with MTBE (1 x 150 mL). The combined organic
layers were dried (Naz;509) and evaporated in vacuo to give a
mixture of compound 19 and benzyl alcohol as an oily white
solid: -H NMR 0300 MH:z, CD30D) b 7.34 (m, 10 H) , 4.63 (s, 2
H), 3.81 (m, 1 H), 3.58 (m, 3 H), 3.03-2.60 (m, 5 H), 2.17
(m, 1 H) , 2.05 ('m, 1 H) , 1.87-1.05 (m, 12 H) , 1.30 (s, 3 H) ,
1.28 (s, 3 H) .
F. Triethylami:ne (3.2 g, 4.3 mL, 31.2 mmol) was added to a
solution of the mixture of amine 19 (4.7 g, 10.4 mmol theory
from 18) and benzyl alcohol in EtOH (23 mL) at ambient
temperature. A solution of 3-acetoxy-2-methylbenzoyl
chloride (20)(obtained according to procedures set forth in
U.S. Patent Application Serial No. 08/708,411, filed
September 5, 1995, which is specifically incorporated by
reference herein) (2.4 g, 11.5 mmol) in THF (4 mL) was
added. After 2 :hours 50% aqueous NaOH (4.1 g, 2.8 mL, 52.2
mmol) was added ~~nd the mixture was heated under reflux for
1 hour. After c~~oling to ambient temperature, the mixture
was neutralized to pH = 7 with 2 N aqueous HC1 (26 mL).
This mixture was diluted with EtOAc (500 mL) and washed with
H20 (1 x 250 mL) , satuo~ated aqueous NaHC03 (2 x 250 mL) , Hz0
(1 x 250 mL), and brine (1 x 125 mL). The organic layer was
dried (Na~S09) and purified by flash chromatography {75/25
EtOAc/hexanes) to give amide 21 as a white foam {1173-57A,
1 . 3 9 g, 23 % ) . T:ze 1H t~TMR indicated the presence of 11 wt
EtOAc which coul~3 not be removed in vacuo.
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Analysis:
1H NMR (300 MHz, CD~OD) S 7.53 (d, J = 7.3 Hz, 2 H} , 7.32 (t,
J = 7.0 Hz, 2 H), 7.20 (t, J = 7.3 Hz, 1 H}, 7.06 (t, J =
8.1 Hz, 1 H}, 6.92 (d, J = 8.1 Hz, 1 H), 6.83 (d, J = 8.1
Hz, 1 H), 4.42 (m, 1 H), 4.08 (m, 1 H}, 3.61 (dd, J = 13.6,
4. 0 Hz, 1 H) , 3 .45 (AB, JAB = 11 .0 Hz, ~uAg = 18.0 Hz, 2 H) ,
3.29 (dd, J = 13.6, 10.3 Hz, 1 H), 3.10 (m, 1 H), 2.66 (m, 2
H), 2.28 (s, 3 H), 2.22 (m, 2 H), 2.04 (m, 1 H), 1.86-1.20
(m, 11 H) , 1.19 (s, 3 H) , 1 .18 (s, 3 H) .
1'C NMR (75.5 MHz, CD30D) ~ 175.7, 172.5, 155.9, 138.8,
136.7, 129.8, 128.9, 126.3, 126.0, 122.4, 118.4, 115.9,
70.3, 69.9, 68.2, 59.3, 58.8, 54.9, 53.0, 36.5, 34.2, 34.1,
31.1, 30.7, 26.4, 26.0, 23.1, 23.0, 20.8, 12.1.
Example 2
HIV Protease inhibition activity and anti HIV activity in
cell culture of compound 21
Tight binding kinetics analysis was used to determine
the magnitude of the Ki values of compound 21. The Ki= 5.6 ~
0.91nM.
Methods
Expression of HIV-1 protease
HIV-1 protease gene was isolated from the viral strain
IIIB (Ratner, L. et al., Nature, 316, 227-284 (1985)). In
order to increase the stability of purified protease (Rose,
J.R, et al., J. Biol. Chem., 268, 11939-11945 (1993)), the
glutamine residue at position 7 (Q7) was mutated to serine
(S) by replacing the 33 base pairs segment between the NdeI
and BstEII sites of the protease gene sequence with
synthetic oligonucleotides encoding the Q75 mutation. The
modified gene sequence was inserted into the plasmid vector
pGZ (Menge, K.L. et la., Biochemistry, 34:15934-15942 (1995)
under the control of phage T7 promoter. The resulting
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construct, pGZ/HP-19Q'7S#9, was transformed into E. coli
strain BL21(DE3) purchased from Novagen, Inc.
Expression of HIV-1 PR: Cultures were grown in 2YT
media (1.6% Try>ticase Pepton, 1% Yeast extract, 0.5 % NaCl
at an initial pH 7.5) containing 200 ~g/L ampicillin in 100
L fermentor (Biolafitt:e SA) at 37°C for 5 hours and then
induced by addition of: 1 mM IPTG {Isopropyl-(3-D-
thiogalactopyran.oside). The temperature of the culture
during induction. was raised to 42°C to increase
accumulations of the recombinant HIV-1 protease as insoluble
inclusion bodies. After 2 hours at 42°C, cells were
harvested by crcssflow filtration using Pellicon 0.1 ~,m
VVPPOOOC5 cassette #10 {Millipore) and the cell paste was
stored frozen at -70°C'.
Purification of Recombinant HIV-1 Protease: All steps
unless otherwise indicated were carried out at 4°C. Protein
concentrations were dE:termined using BioRad protein assay
solution with bovine ~;erum albumin (BioRad, Richmond, CA) as
a standard. Chromatographic steps and the purity of HIV PR
was analyzed by sodium dodecylsulfate polyacrylamide gel
electrophoresis (SDS-PAGE). Final purity of HIV-PR was
> 98o. Typical final yield from each 100 L culture was 120
mg.
Cell paste from 1.00L culture was resuspended in 300 mL
of lysis buffer (50 mM Tris-C1 pH 8.0, 25 mM NaCl, 20 mM 2-
mercaptoethanol) and microfluidized in Microfluidics
Corporation fluidizer at 22,000 psi. The crude cell lysate
was clarified by centrifugation at 14,000 rpm for 20
minutes. HIV PR was found predominantly in the pellet in the
form of inclusion bodies. The inclusion bodies were
subsequently washed multiple times in the lysis buffer
containing in addition 0.1% Trition-X100 and 1 M urea, and
after each washing procedure, the inclusion bodies were
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pelleted by centrifugation at 5,000 rpm for 20 minutes.
Purified inclusion bodies were solubilized in buffer
containing 50 mM Tris-C1, pH 8.0, 25 mM NaCl, 20 mM 2-
mercaptoethanol, and 8 M urea. Solution was clarified by
centrifugation at 14,000 rpm and applied at room temperature
to a 300 mL Fast Flow Q-Sepharose column (Pharmacia,
Piscataway, NJ) equilibrated with the same buffer. Under
these conditions HIV PR did not bind to the column and
essentially pure enzyme was found in the flow-through
fractions. To renature the protein, the fractions from Fast
Flow Q-Sepharose column were dialyzed against three changes
of buffer containing 25 mM NaH2P04 pH 7.0, 25 mM NaCl, 10 mM
DTT and 10o glycerol. After refolding, small quantities of
precipitated material were removed by centrifugation and
resultant enzyme preparation were concentrated, dialyzed
against 0.5 M NaCl, 50 mM MES pH 5.6, 10 mM DTT, frozen in
small aliquots at "'2 mg/mL and stored at -70°.
Tight-Binding Kinetics Assay and Analysis
Proteolytic activity of purified HIV-1 protease was
measured using a modified chromogenic assay developed by
Richards at al. (Richards, A.D. et al. J. Biol. Chem., 256,
773-7736 (1990)). The synthetic peptide His-Lys-Ala-Arg-
Val-Leu-Phe(paraN02)-Glu-Ala-Nle-Ser-NHz (American Peptide
Company) (Nle is norleucine) was used as a substrate. The
assay was carried out in 0.5 M NaCl, 50 mM MES pH 5.6, 5 mM
DDT, and 2o DMSO at 37°C. Cleavage of the scissile bond
between leucine and paranitro-phenylalanine (Phe para-N02)
was assayed by spectrophotometric monitoring of the decrease
on absorbance at 305 nm. Initial velocity was determined as
the rate of decline of absorbance during the first 100
seconds of the enzymatic reaction. Under these conditions,
and using Q7S HIV-1 protease, the Michaelis constant (Km)
for this substrate is 59 ~ 17 /.cM.
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For determ:inatio:n of the inhibition of compound 21, a
saturating concE:ntration of substrate of 200 uM was used.
Between 13 and 20 con~~entrations of inhibitors were
evaluated and the velocity of reaction was measured at each
concentration a:~ desc:ribed above. The apparent Ki (Ki app),
set forth above,. was determined by computer assisted non-
linear least square fitting of the data to the tight binding
equation of Mor~:ison (Morrison, J.F., Biochem. Biophys.
Acta, 185, 269-~?86 (1963) ) .
Example 3
Antiviral activity of compound 21 against HIV-1 in cell
culture
Cells and virus strains:
The CEM-SS and MT-2 human T cell lines and HIV-1
strains RF and 7:IIB we=re obtained from the AIDS Research and
Reference Progr~im, Division of AIDS, NIAID, and NIH.
Cell protection assavs_
The inhibitory ei=fects of each agent on HIV-1
replication were measured by the MTT dye reduction method
(Alley, M.C. et al., Cancer Res. 48: 589-601 (1988)).
Compounds were dissolved in DMSO at a concentration of
40 mg/ml then diluted 1:200 in culture medium (RPMI,
supplemented with 10% fetal bovine serum). From each
diluted stock, 1.00 ~1 was added to a 96-well plate and
serial half-log dilutions were prepared. In separate tubes,
MT-2 cells and C'EM-SS cells were infected with HIV-1 IIIB or
HIV-1 RF at a multiplicity of infection (m.o.i.) of 0.01 and
0.03, respectively. Following a 4-hour adsorption period,
100 ~.1 of infected or uninfected cells were added to the .
wells of the drug containing plate to give a final
concentration of 1 x L09 cells/well. Six days (CEM-SS
cells) or 7 days; (MT-2 cells) later, MTT (5 mg/ml) was added
to test plates and the amount of formazan produced was
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quantified spectrophotometrically at 570 nm. Data were
expressed as the percentage of formazan produced in drug-
treated cells compared to formazan produced in wells of
uninfected, drug-free cells. The EDSO was calculated as the
concentration of drug that increased the percentage of
formazan production in infected, drug-treated cells to 500
of that produced by uninfected, drug-free cells.
Cytotoxicity (TCSp) was calculated as the concentration of
drug that decreased the percentage of formazan produced in
uninfected, drug-treated cells to 50% of that produced in
uninfected, drug-free cells. The therapeutic index (TI) was
calculated by dividing the cytotoxicity (TCSO) by the
antiviral efficacy (ED50).
Table 1
Antiviral Activity and Cytotoxicity Evaluations of Compound
21
in an Acute Infection of CEM-SS cells with HIV-1 RF
Compound ED50 ED95 TC50 Therapeutic
(nM) (nM) (~M) index a
21 34.2 154.1 96.6 2825
azidophymidine 52.3 543.1 >374.5 >7161
(AZT)
dideoxycytidine 94.70 142.0 37.69 398
(ddC)
Therapeutic index = Cytotoxicity (TCS~) . Antiviral
activity (EDSO) -
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Table 2
Antiviral Activity and Cytotoxicity Evaluations of Compound
21
in an Acute Infection of MT-2 cells with HIV-1 IIIB
Compound ED~~~ ED93 TC50 Therapeu
(nM) (nM) (~,M) tic
21 85.6 ND 92.6 1082
AZT 430.7 ND 109.4 254
ddC 5924 ND 176.3 30
aTherapeutic index = C'ytotoxicity (TCSp) . Antiviral
activity (ED50) .
As noted ar~ove, t:he compounds of the present invention
are useful for inhibit:ing HIV protease, which is an enzyme
associated with viral component production and assembly. An
embodiment of th.e pre~~ent invention is a method of treating
HIV infection comprising administering to a host or patient,
such as a primate, an effective amount of a compound of
formula (9) or a pharmaceutically acceptable salt thereof.
Another embodiment of the present invention is a method of
treating AIDS comprising administering to a host or patient
an effective amount of a compound of formula (9) or a
pharmaceutically acceptable salt thereof. A further
embodiment of the pre~~ent invention is a method of
inhibiting HIV proteae;e comprising administering to an HIV
infected cell or a ho~;t or patient, such as a primate,
infected with HIV, an effective amount of a compound of
formula (1) or a pharmaceutically acceptable salt thereof.
The term "effecti.ve amount" means an amount of a
compound of formula (9) or its pharmaceutically acceptable
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salt that is effective to inhibit the HIV protease mediated
viral component production and assembly. The specific dose
of compound administered according to this invention to
obtain therapeutic or inhibitory effects will, of course, be
determined by the particular circumstances surrounding the
case, including, for example, the compound administered, the
route of administration, the condition being treated and
the individual host or patient being treated. An exemplary
daily dose (administered in single or divided doses)
contains a dosage level of from about 0.01 mg/kg to about 50
mg/kg of body weight of a compound of this invention.
Preferred daily doses generally are from about 0.05 mg/kg to
about 40 mg/kg and, more preferably, from about 1.0 mg/kg to
about 30 mg/kg.
The compounds of the invention may be administered by a
variety of routes, including oral, rectal, transdermal,
subcutaneous, intravenous, intramuscular and intranasal
routes. The compounds of the present invention are
preferably formulated prior to administration. Therefore,
another embodiment of the present invention is a
pharmaceutical composition or formulation comprising an
effective amount of a compound of formula (9) or a
pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier, such as a diluent or
excipient therefor.
The active ingredient preferably comprises from 0.1% to
99.9% by weight of the formulation. By "pharmaceutically
acceptable" it is meant that the carrier, such as the
diluent or excipient, is compatible with the other
ingredients of the formulation and not deleterious to the
host or patient.
Pharmaceutical formulations may be prepared from the
compounds of the invention by known procedures using known-
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and readily available ingredients. In making the
compositions of the present invention, the active ingredient
will usually be admixed with a carrier, or diluted by a
carrier, or enclosed within a carrier, which may be in the
form of a capsu=.e, sa~~het, paper or other suitable
container. 49hen the carrier serves as a diluent, it may be
a solid, semi-solid o:r liquid material which acts as a
vehicle, excipient or medium for the active ingredient.
Thus, the compo:~itions can be in the form of tablets, pills,
powders, lozenge's, sachets, cachets, elixirs, suspensions,
emulsions, solutions, syrups, aerosols (as a solid or in a
liquid medium), ointmf_nts (containing, for example, up to
loo by weight of: the <~ctive compound), soft and hard gelatin
capsules, suppo~~itories, sterile injectable solutions,
sterile packaged powders and the like.
The following formulation examples are illustrative
only and are not; intended to limit the scope of the
invention. The term "active ingredient" represents a
compound of formula (9) or a pharmaceutically acceptable
salt thereof.
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Formulation 1
Hard gelatin capsules are prepared using the following
ingredients:
Quantity
(m~/capsule)
Active ingredient 250
Starch, dried 200
Magnesium stearate 10
Total 460 mg
Formulation 2
A tablet is prepared using the ingredients below:
Quantity
(mc~/tablet)
Active ingredient 250
Cellulose, microcrystalline 400
Silicon dioxide, fumed 10
Stearic acid 5
Total 665 mg
The components are blended and compressed to form tablets
each weighing 665 mg.
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Formulation 3
An aerosol so:Lution is prepared containing the following
components:
Weight
Active ingredient 0.25
Methanol 25.75
Propellant 22
(Chlorodifluoromethane) 74.00
Tota 100.00
The active compound is mixed with ethanol and the mixture
added to a portion of tl:~e propellant 22, cooled to -30°C and
transferred to a i=filling device. The required amount is then
fed to a stainles:~ steel container and diluted with the
remainder of the propel:Lant. The valve units are then fitted
to the container.
Formulation 4
Tablets, each containing 60 mg of active ingredient, are
made as follows:
Quantity
(mg/tablet)
Active ingredient 60
Starch 45
Microcrystalline cellulose 35
Polyvinylpyrrolidone
(as loo solution in waiver) 4
Sodium carboxymethyl sta=rch 4.5
Magnesium stearate 0.5
Talc 1
Total 150
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The active ingredient, starch and cellulose are passed through
a No. 45 mesh U.S. sieve and mixed thoroughly. The aqueous
solution containing polyvinylpyrrolidone is mixed with the
resultant powder, and the mixture then is passed through a No.
14 mesh U.S. sieve. The granules so produced are dried at 50°C
and passed through a No. 18 mesh U.S. sieve. The sodium
carboxymethyl starch, magnesium stearate and talc, previously
passed through a No. 60 mesh U.S. sieve, are then added to the
granules which, after mixing, are compressed on a tablet
machine to yield tablets each weighing 150 mg.
Formulation 5
Capsules, each containing 80 mg of active ingredient, are
made as follows:
Quantity
(mg/capsule)
Active ingredient 80 mg
Starch 59 mg
Microcrystalline cellulose 59 mg
Magnesium stearate 2 mg
Total 200 mg
The active ingredient, cellulose, starch and magnesium
stearate are blended, passed through a No. 45 mesh U.S. sieve,
and filled into hard gelatin capsules in 200 mg quantities.
Formulation 6
Suppositories, each containing 225 mg of active
ingredient, are made as follows:
Active ingredient 225 mg
Saturated fatty acid glycerides 2.000 mg
Total 2,225 mg
The active ingredient is passed through a No. 60 mesh U.S.
sieve and suspended in the saturated fatty acid glycerides
previously melted using the minimum heat necessary. The
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mixture is then poured into a suppository mold of nominal 2 g
capacity and allowed to cool.
Formulation 7
Suspensions, a>.ach containing 50 mg of active ingredient
per 5 ml dose, are made as follows:
Active ingredient 50 mg
Sodium carboxymethyl cellulose 50 mg
Syrup 1.25 ml
Benzoic acid solution 0.10 ml
Flavor q,v,
Color q.v,
Purified water to total 5 ml
The active ingredient is passed through a No. 45 mesh U.S.
sieve and mixed with the sodium carboxymethylcellulose and
syrup to form a smooth paste. The benzoic acid solution,
flavor and color are diluted with a portion of the water and
added, with stirring. Sui=ficient water is then added to
produce the requirE~d volume .
Formulation 8
An intravenou~o formulation is prepared as follows:
Active ingredient 100 mg
Isotonic saline 1,00() mL
The solution of the: above ingredients generally is
administered intravenously to a subject at a rate of 1 ml per
minute.
SUBSTITUTE SHEET (RULE 26)
CA 02284163 1999-09-10
WO 98/40357 PCT/US98/04735
-46-
Formulation 9
A tablet is prepared using the ingredients below:
Quantity
(ma/tablet)
Active ingredient 292 mg
calcium silicate 146 mg
crospovidone 146 mg
Magnesium stearate 5 mg
Total 589 mg
SUBSTITUTE SHEET (RULE 26)
