U.S. patent application number 12/684845 was filed with the patent office on 2010-05-27 for anticonvulsant and anxiolytic methods of using receptor subtype selective agents.
Invention is credited to James M. Cook, Dongmel Han, Xiaohui He, Qi Huang, Xioeyan Li, Jianming Yu.
Application Number | 20100130481 12/684845 |
Document ID | / |
Family ID | 28675480 |
Filed Date | 2010-05-27 |
United States Patent
Application |
20100130481 |
Kind Code |
A1 |
Cook; James M. ; et
al. |
May 27, 2010 |
ANTICONVULSANT AND ANXIOLYTIC METHODS OF USING RECEPTOR SUBTYPE
SELECTIVE AGENTS
Abstract
Methods of using orally active benzodiazepine derivatives and
their salts are disclosed. These compounds and their salts have
anticonvulsant and anxiolytic activity with reduced sedative,
hypnotic, muscle relaxant, or ataxic effects.
Inventors: |
Cook; James M.; (US)
; Huang; Qi; (US) ; He; Xiaohui; (US)
; Li; Xioeyan; (US) ; Yu; Jianming;
(US) ; Han; Dongmel; (US) |
Correspondence
Address: |
WISYS TECHNOLOGY FOUNDATION, INC.;c/o MIRICK, O'CONNELL, DEMALLIE &
LOUGEE, LLP
1700 WEST PARK DRIVE/ ROGER P. ZIMMERMAN, ESQ.
WESTBOROUGH
MA
01581-3941
US
|
Family ID: |
28675480 |
Appl. No.: |
12/684845 |
Filed: |
January 8, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11929860 |
Oct 30, 2007 |
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12684845 |
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11458855 |
Jul 20, 2006 |
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11929860 |
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10402538 |
Mar 28, 2003 |
7119196 |
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11458855 |
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60368408 |
Mar 28, 2002 |
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Current U.S.
Class: |
514/220 |
Current CPC
Class: |
C07D 487/04 20130101;
C07D 495/14 20130101; A61P 25/00 20180101; C07D 243/20 20130101;
C07D 243/12 20130101; A61P 25/18 20180101; C07D 487/06 20130101;
C07D 519/00 20130101; A61P 1/08 20180101; A61P 25/22 20180101; C07D
495/04 20130101; C07D 243/24 20130101; C07D 471/04 20130101; C07D
498/04 20130101 |
Class at
Publication: |
514/220 |
International
Class: |
A61K 31/551 20060101
A61K031/551; A61P 25/22 20060101 A61P025/22 |
Goverment Interests
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0002] This invention was made with Government support under NIMH
grant number MH46851. The Government has certain rights to this
invention.
Claims
1. A method for the treatment and/or prevention of convulsions
comprising: administering to a mammalian subject in need of such
treatment an effective amount of a compound of formula VIII, or a
salt thereof, ##STR00078## wherein X is N or CH; Y and Z are taken
together with the two intervening carbon atoms to form a ring
selected from phenyl and thienyl, which ring is substituted at the
C(8) position with at least the substituent --C.ident.C--R, where R
is H, Si(CH.sub.3).sub.3, t-butyl, isopropyl, methyl, or
cyclopropyl; R.sub.1 is H, CH.sub.3, CF.sub.3, CH.sub.2CH.sub.3,
CH.sub.2CF.sub.3, or cyclopropyl; R.sub.2 is a substituted or
unsubstituted at least partially unsaturated 5 or 6 membered cyclic
or heterocyclic ring, wherein if substituted, the substituent is F,
Cl, Br, or NO.sub.2 at the 2'-position.
2. The method according to claim 1, wherein the compound is
##STR00079## ##STR00080## wherein R is H or Si(CH.sub.3).sub.3, R'
is H or CH.sub.3, and X' is H, F, Cl, Br, or NO.sub.2.
3. The method according to claim 1, wherein the compound is
##STR00081##
4. A method for the treatment and/or prevention of anxiety
comprising: administering to a mammalian subject in need of such
treatment an effective amount of a compound of formula VIII, or a
salt thereof, ##STR00082## wherein: X is N or CH; Y and Z are taken
together with the two intervening carbon atoms to form a ring
selected from phenyl and thienyl, which ring is substituted at the
C(8) position with at least the substituent --C.ident.C--R, where R
is H, Si(CH.sub.3).sub.3, t-butyl, isopropyl, methyl, or
cyclopropyl; R.sub.1 is H, CH.sub.3, CF.sub.3, CH.sub.2CH.sub.3,
CH.sub.2CF.sub.3, or cyclopropyl; R.sub.2 is a substituted or
unsubstituted at least partially unsaturated 5 or 6 membered cyclic
or heterocyclic ring, wherein if substituted, the substituent is F,
Cl, Br, or NO.sub.2 at the 2'-position.
5. The method according to claim 4, wherein the compound is
##STR00083## ##STR00084## wherein R is H or Si(CH.sub.3).sub.3, R'
is H or CH.sub.3, and X' is H, F, Cl, Br, or NO.sub.2.
6. The method according to claim 4, wherein the compound is
##STR00085##
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation application, and claims
benefit of co-pending U.S. patent application Ser. No. 11/929,860,
filed on Oct. 30, 2007, which is a divisional application and
claims the benefit of U.S. patent application Ser. No. 11/458,855,
filed Jul. 20, 2006, now abandoned, which is a continuation of U.S.
patent application Ser. No. 10/402,538, filed on Mar. 28, 2003, now
U.S. Pat. No. 7,119,196, which claims benefit of U.S. Provisional
Application No. 60/368,408, filed Mar. 28, 2002, all of which are
incorporated herein in their entirety as if fully set forth
herein.
BACKGROUND OF THE INVENTION
[0003] The present invention relates to a class of benzodiazepine
derivatives which possess anxiolytic activity with decreased
sedative, hypnotic, and ataxic side effects.
[0004] The most frequently prescribed medication for treatment of
anxiety disorders (such as phobias, obsessive compulsive disorders)
and seizure disorders are benzodiazepines such as diazepam
(VALIUM.RTM.), triazolam (HALCION.RTM.), midazolam (VERSED.RTM.),
lorazepam (ATIVAN.RTM.), chlordiazepoxide (LIBRIUM.RTM.),
alprazolam (XANAX.RTM.), and other benzodiazepine-based
medications. However, these benzodiazepine-based medications have
side effects such as drowsiness, sedation, motor incoordination,
memory impairment, potentiation of effects of alcohol, tolerance
and dependence, and abuse potential. Buspirone, tandospirone, and
other serotonergic agents have been developed as anxiolytics with a
potentially reduced profile of side effects. However, while these
medications do show a reduced profile of side effects, they have
other characteristics which make them less than ideal for treatment
of anxiety disorders. In some cases, these agents cause anxiety
before a therapeutic dose can be obtained or require dosing of the
drug for several days before a therapeutic effect is seen.
Development of anxiolytics with even fewer side effects is
desired.
[0005] Receptors for the major inhibitory neurotransmitter,
gamma-aminobutyric acid (GABA), are divided into three main
classes: (1) GABA.sub.A receptors, which are members of the
ligand-gated ion channel superfamily; (2) GABA.sub.B receptors,
which may be members of the G-protein linked receptor superfamily;
and (3) GABA.sub.C receptors, also members of the ligand-gated ion
channel superfamily, but their distribution is confined to the
retina. Benzodiazepine receptor ligands do not bind to GABA.sub.B
and GABA.sub.C receptors. Since the first cDNAs encoding individual
GABA.sub.A receptor subunits were cloned the number of known
members of the mammalian family has grown to 21 including .alpha.,
.beta., and .gamma. subunits (6.alpha., 4.beta., 4.gamma.,
1.delta., 1.epsilon., 1.pi., 1.theta., and 3.rho.).
[0006] Subtype assemblies containing an .alpha.1 subunit
(.alpha.1.beta.2.gamma.2) are present in most areas of the brain
and are thought to account for 40-50% of GABA.sub.A receptors in
the rat. Subtype assemblies containing .alpha.2 and .alpha.3
subunits respectively are thought to account for about 25% and 17%
GABA.sub.A receptors in the rat. Subtype assemblies containing an
.alpha.5 subunit (.alpha.5.beta.3.gamma.2) are expressed
predominately in the hippocampus and cortex and are thought to
represent about 4% of GABA.sub.A receptors in the rat.
[0007] A characteristic property of all known GABA.sub.A receptors
is the presence of a number of modulatory sites, one of which is
the benzodiazepine binding site. The benzodiazepine binding site is
the most explored of the GABA.sub.A receptor modulatory sites, and
is the site through which benzodiazepine-based anxiolytic drugs
exert their effect. Before the cloning of the GABA.sub.A receptor
gene family, the benzodiazepine binding site was historically
subdivided into two subtypes, BENZODIAZEPINE1 and BENZODIAZEPINE2,
on the basis of radioligand binding studies on synaptosomal rat
membranes. The BENZODIAZEPINE1 subtype has been shown to be
pharmacologically equivalent to a GABA.sub.A receptor comprising
the .alpha.1 subunit in combination with a .beta. subunit and
.gamma.2. This is the most abundant GABA.sub.A receptor subtype,
and is believed to represent almost half of all GABA.sub.A
receptors in the brain, as stated.
[0008] Two other major populations are the
.alpha.2.beta.2/3.gamma.2 and .alpha.3.beta.2/3.gamma.2/3 subtypes.
Together these constitute approximately a further 35% of the total
GABA.sub.A receptor population. Pharmacologically this combination
appears to be equivalent to the BENZODIAZEPINE2 subtype as defined
previously by radioligand binding, although the BENZODIAZEPINE2
subtype may also include certain .alpha.5-containing subtype
assemblies. The physiological role of these subtypes has hitherto
been unclear because no sufficiently selective agonists or
antagonists were known.
[0009] It is now believed that agents acting as benzodiazepine
agonists at GABA.sub.A/.alpha.2, GABA.sub.A/.alpha.3, and/or
GABA.sub.A/.alpha.5 receptors, will possess desirable anxiolytic
properties. Compounds which are modulators of the benzodiazepine
binding site of the GABA.sub.A receptor by acting as benzodiazepine
agonists are referred to hereinafter as "GABA.sub.A receptor
agonists." The GABA.sub.A/.alpha.1-selective
(.alpha.1.beta.2.gamma.2) agonists alpidem and zolpidem are
clinically prescribed as hypnotic agents, suggesting that at least
some of the sedation associated with known anxiolytic drugs which
act at the BENZODIAZEPINE1 binding site is mediated through
GABA.sub.A receptors containing the al subunit. Accordingly, it is
considered that GABA.sub.A/.alpha.2, GABA.sub.A/.alpha.3, and/or
GABA.sub.A/.alpha.5 receptor agonists rather than
GABA.sub.A/.alpha.1 receptors will be effective in the treatment of
anxiety with a reduced propensity to cause sedation. For example,
QH-ii-066 binds with high affinity to GABA.sub.A/.alpha.5 receptors
(Ki<10 nM), intermediate affinity to GABA.sub.A/.alpha.2 and
GABA.sub.A/.alpha.3 (Ki<50 nM), and lower affinity to
GABA.sub.A/.alpha.1 receptors (Ki>70 nM), unlike diazepam which
binds with high affinity to all four diazepam-sensitive GABA.sub.A
receptors (Ki<25 nM), as disclosed in Huang, et al., J. Med.
Chem. 2000, 43, 71-95. Also, agents which are antagonists or
inverse agonists at .alpha.1 receptors might be employed to reverse
sedation or hypnosis caused by .alpha.1 agonists.
[0010] Since the compounds of the present invention exhibit
increased agonist efficacy at only a few GABA.sub.A types of
receptors and/or selective efficacy at one or more ion channels and
have been shown to be effective in animal models of anxiety and
seizures, with reduced severity and/or incidence of side effects,
they are useful in the treatment and/or prevention of a variety of
disorders of the central nervous system. Such disorders include
anxiety disorders, such as panic disorder with or without
agoraphobia, agoraphobia without history of panic disorder, animal
and other phobias including social phobias, obsessive-compulsive
disorder, general anxiety disorder, attention deficit disorders,
stress disorders including post-traumatic and acute stress
disorder, and generalized or substance-induced anxiety disorder,
neuroses, convulsions; migraine; depressive or bipolar disorders,
for example single episode or recurrent major depressive disorder,
dysthymic disorder, bipolar I and bipolar II manic disorders, and
cyclothymic disorder, psychotic disorders including
schizophrenia.
SUMMARY OF THE INVENTION
[0011] In consideration of this situation, the problem to be solved
by the present invention is to provide a medication which can be
used for the treatment of anxiety neurosis, general anxiety
disorder, panic disorder, phobias, obsessive-compulsive disorders,
schizophrenia, post-cardiac trauma stress disorders, depression
disorders, psychosomatic disorders, and other psychoneurotic
disorders, eating disorders, menopausal disorders, infantile autism
and other disorders, and also emesis with fewer side effects.
[0012] The present inventors engaged in repeated extensive studies
to develop a superior medication free from the above problems. They
found that the compounds of the present invention, that is, the
novel benzodiazepine derivatives and their salts, have beneficial
pharmacological and behavioral effects, that is, the compounds of
the present invention show anxiolytic and anticonvulsant activity
with greatly decreased or no sedative/hypnotic/muscle
relaxant/ataxic side effects.
[0013] The compounds described in the present invention have been
synthesized based on a modified version of the computer modeling
disclosed in Cook, et al J. Med. Chem., 1996, 39, 1928-1934. These
compounds obtained by modifying elements, described herein, of the
known benzodiazepine agents, have increased binding selectivity for
the GABA.sub.A/.alpha.2, GABA.sub.A/.alpha.3, and/or
GABA.sub.A/.alpha.5 receptors described above, and/or altered
efficacy at one or more GABA.sub.A receptors described above,
and/or altered selectivity at one or more ion channels. These
compounds, which have been tested in animal models of anxiety in
rats and seizures in mice, and side effect models in rats, have
been found to be orally active and have anxiolytic and
anticonvulsant activity, with reduced severity and/or incidence of
side effects.
[0014] One object of the present invention is to identify
medications containing these benzodiazepine derivatives or their
pharmaceutically acceptable salts as essential ingredients that are
usable for the treatment of anxiety neurosis, phobias,
obsessive-compulsive disorders, panic disorder, generalized anxiety
disorder, schizophrenia, post-cardiac trauma stress disorders,
depression disorders, psychosomatic disorders, and other
psychoneurotic disorders, eating disorders, menopausal disorders,
infantile autism, and other disorders.
[0015] The present invention describes a class of benzodiazepine
derivatives which possess desirable enhanced agonist efficacy at
various GABA.sub.A receptors and desirable behavioral profile with
respect to anxiolytic and anticonvulsant efficacy and reduced side
effect efficacy. The compounds in accordance with the present
invention have agonist efficacy at the GABA.sub.A/.alpha.2,
GABA.sub.A/.alpha.3, and GABA.sub.A/.alpha.5 receptors. The
compounds of this invention have anxiolytic and anticonvulsant
effects with decreased sedative-hypnotic activity.
[0016] The present invention provides a compound of formula I, or a
salt or prodrug thereof,
##STR00001##
wherein Y and Z are taken together with the two intervening carbon
atoms to form a ring selected from phenyl and thienyl, which ring
is substituted at the C(7) position with at least the substituent
--C.ident.C--R, where R is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; R.sub.1 is one of H, CH.sub.3,
C.sub.2H.sub.4N(C.sub.2H.sub.5).sub.2, CH.sub.2CF.sub.3,
CH.sub.2C.ident.CH, or an alkyl cyclopropyl; R.sub.2 is a
substituted or unsubstituted at least partially unsaturated 5 or 6
membered cyclic or heterocyclic ring, wherein if substituted the
substituent is one or more of F, Cl, Br, or NO.sub.2 at the
2'-position; and R.sub.3 is one of H, OH, OCON(CH.sub.3).sub.2,
COOCH.sub.3, or COOC.sub.2H.sub.5. Preferred compounds according to
formula I include:
##STR00002##
[0017] The invention provides in another aspect a compound of
formula II, or a salt or prodrug thereof,
##STR00003##
wherein Y and Z are taken together with the two intervening carbon
atoms to form a ring selected from phenyl and thienyl, which ring
is substituted at the C(7) position with at least the substituent
--C.ident.C--R, where R is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; R.sub.1 is one of H, CH.sub.3,
C.sub.2H.sub.4N(C.sub.2H.sub.5).sub.2, CH.sub.2CF.sub.3,
CH.sub.2C.ident.CH, or an alkyl cyclopropyl; and R.sub.2 is a
substituted or unsubstituted at least partially unsaturated 5 or 6
membered cyclic or heterocyclic ring, wherein if substituted the
substituent is one or more of F, Cl, Br, or NO.sub.2 at the
2'-position. Preferred compounds according to formula II
include:
##STR00004##
[0018] The present invention provides in yet another aspect a
compound of formula III, or a salt or prodrug thereof,
##STR00005##
wherein Y and Z are taken together with the two intervening carbon
atoms to form a ring selected from phenyl and thienyl, which ring
is substituted at the C(7) position with at least the substituent
--C.ident.C--R, where R is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; and R.sub.2 is a substituted or
unsubstituted at least partially unsaturated 5 or 6 membered cyclic
or heterocyclic ring, wherein if substituted the substituent is one
or more of F, Cl, Br, or NO.sub.2 at the 2'-position. Preferred
compounds according to the formula III include:
##STR00006##
[0019] Further, the present invention provides a compound of
formula IV, or a salt or prodrug thereof,
##STR00007##
wherein R is H, Si(CH.sub.3).sub.3, t-butyl, isopropyl, methyl, or
cyclopropyl; R.sub.1 is one of H, CH.sub.3, C.sub.2H.sub.4N
(C.sub.2H.sub.5).sub.2, CH.sub.2CF.sub.3, CH.sub.2C.ident.CH, or an
alkyl cyclopropyl; R.sub.2 is a substituted or unsubstituted at
least partially unsaturated 5 or 6 membered cyclic or heterocyclic
ring, wherein if substituted the substituent is one or more of F,
Cl, Br, or NO.sub.2 at the 2'-position; and A is an ethoxide or a
propoxide. Preferred compounds according to the formula IV
include:
##STR00008##
[0020] In a still further aspect, the present invention provides a
compound of formula V, or a salt or prodrug thereof,
##STR00009##
wherein Y and Z are taken together with the two intervening carbon
atoms to form a ring selected from phenyl and thienyl, which ring
is substituted at the C(8) position with at least the substituent
--C.ident.C--R, where R is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; R.sub.1 is one of H, CH.sub.3,
CF.sub.3, CH.sub.2CH.sub.3, CH.sub.2CF.sub.3, CH.sub.2C.ident.CH,
an alkyl, or cyclopropyl; R.sub.2 is a substituted or unsubstituted
at least partially unsaturated 5 or 6 membered cyclic or
heterocyclic ring, wherein if substituted the substituent is one or
more of F, Cl, Br, or NO.sub.2 at the 2'-position; and R.sub.5 is a
branched or straight chain C.sub.1 to C.sub.4 halogenated or
unhalogenated alkyl or a methyl cyclopropyl. Preferred compounds
according to formula V include:
##STR00010##
[0021] In yet another aspect, the present invention provides a
compound of formula VI, or a salt or prodrug thereof,
##STR00011##
wherein Y and Z are taken together with the two intervening carbon
atoms to form a ring selected from phenyl and thienyl, which ring
is substituted at the C(8) position with at least the substituent
--C.ident.C--R, where R is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; R.sub.1 is one of H, CH.sub.3,
CF.sub.3, CH.sub.2CH.sub.3, CH.sub.2CF.sub.3, or cyclopropyl;
R.sub.2 is a substituted or unsubstituted at least partially
unsaturated 5 or 6 membered cyclic or heterocyclic ring, wherein if
substituted the substituent is one or more of F, Cl, Br, or
NO.sub.2 at the 2'-position; and R.sub.6 is a branched or straight
chain C.sub.1 to C.sub.4 alkyl or a methyl cyclopropyl. Preferred
compounds according to formula VI include:
##STR00012##
[0022] The present invention also provides a compound of formula
VII, or a salt or prodrug thereof,
##STR00013##
wherein Y and Z are taken together with the two intervening carbon
atoms to form a ring selected from phenyl and thienyl, which ring
is substituted at the C(8) position with at least the substituent
--C.ident.C--R, where R is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; and R.sub.2 is a substituted or
unsubstituted at least partially unsaturated 5 or 6 membered cyclic
or heterocyclic ring, wherein if substituted the substituent is one
or more of F, Cl, Br, or NO.sub.2 at the 2'-position. Preferred
compounds according to formula VII include:
##STR00014##
[0023] The present invention still further provides a compound of
formula VIII, or a salt or prodrug thereof,
##STR00015##
wherein Y and Z are taken together with the two intervening carbon
atoms to form a ring selected from phenyl and thienyl, which ring
is substituted at the C(8) position with at least the substituent
--C.ident.C--R, where X is N or CH, where R is H,
Si(CH.sub.3).sub.3, t-butyl, isopropyl, methyl, or cyclopropyl;
R.sub.1 is H, CH.sub.3, CF.sub.3, CH.sub.2CH.sub.3,
CH.sub.2CF.sub.3, or cyclopropyl; R.sub.2 is a substituted or
unsubstituted at least partially unsaturated 5 or 6 membered cyclic
or heterocyclic ring, wherein if substituted the substituent is one
or more of F, Cl, Br, or NO.sub.2 at the 2'-position. Preferred
compounds according to formula VIII include:
##STR00016##
[0024] Yet another aspect of the present invention provides a
compound of formula IX, or a salt or prodrug thereof,
##STR00017##
wherein n is 0 to 4; Y and Z are taken together with the two
intervening carbon atoms to form a ring selected from phenyl and
thienyl, which ring is substituted at the C(8) position with at
least the substituent --C.ident.C--R, where R is H,
Si(CH.sub.3).sub.3, t-butyl, isopropyl, methyl, or cyclopropyl; Y'
and Z' are taken together with the two intervening carbon atoms to
form a ring selected from phenyl and thienyl, which ring is
substituted at the C(8)' position with at least the substituent
--C.ident.C--R', where R' is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; R.sub.1 and R.sub.1' are
independently one of H, CH.sub.3, CF.sub.3, CH.sub.2CF.sub.3,
CH.sub.2CH.sub.3, or cyclopropyl; and R.sub.2 and R.sub.2' are
independently a substituted or unsubstituted at least partially
unsaturated 5 or 6 membered cyclic or heterocyclic ring, wherein if
substituted the substituent is one or more of F, Cl, Br, or
NO.sub.2 at the 2'-position. Preferred compounds according to
formula IX include:
##STR00018##
[0025] A still further aspect of the present invention provides a
compound of formula X, or a salt or prodrug thereof,
##STR00019##
wherein Y and Z are taken together with the two intervening carbon
atoms to form a ring selected from phenyl and thienyl, which ring
is substituted at the C(8) position with at least the substituent
--C.ident.C--R, where R is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; Y' and Z' are taken together
with the two intervening carbon atoms to form a ring selected from
phenyl and thienyl, which ring is substituted at the C(8)' position
with at least the substituent --C.ident.C--R' where R' is H,
Si(CH.sub.3).sub.3, t-butyl, isopropyl, methyl, or cyclopropyl;
R.sub.1 and R.sub.1' are independently one of H, CH.sub.3,
CF.sub.3, CH.sub.2CH.sub.3, CH.sub.2CF.sub.3, or cyclopropyl;
R.sub.2 and R.sub.2' are independently a substituted or
unsubstituted at least partially unsaturated 5 or 6 membered cyclic
or heterocyclic ring, wherein if substituted the substituent is one
or more of F, Cl, Br, or NO.sub.2 at the 2'-position; and B is O or
NH and wherein --BCH.sub.2B-- is optionally replaced with
--N(R.sub.7)--N(R.sub.7)--, where R.sub.7 is one of H, CH.sub.3,
alkyl, or cycloalkyl. Preferred compounds according to formula X
include:
##STR00020##
[0026] The present invention further provides a compound of formula
XI, or a salt or prodrug thereof,
##STR00021##
wherein n is 1 or 2; wherein Y and Z are taken together with the
two intervening carbon atoms to form a ring selected from phenyl
and thienyl, which ring is substituted at the C(8) position with at
least the substituent --C.ident.C--R, where R is H,
Si(CH.sub.3).sub.3, t-butyl, isopropyl, methyl, or cyclopropyl; Y'
and Z' are taken together with the two intervening carbon atoms to
form a ring selected from phenyl and thienyl, which ring is
substituted at the C(8)' position with at least the substituent
--C.ident.C--R', where R' is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; R.sub.1 and R.sub.1' are
independently one of H, CH.sub.3, CF.sub.3, CH.sub.2CH.sub.3,
CH.sub.2CF.sub.3, or cyclopropyl; R.sub.2 and R.sub.2' are
independently a substituted or unsubstituted at least partially
unsaturated 5 or 6 membered cyclic or heterocyclic ring, wherein if
substituted the substituent is one or more of F, Cl, Br, or
NO.sub.2 at the 2'-position; and B is O, NH, or
--N(R.sub.7)--N(R.sub.7)--, where R.sub.7 is one of H, CH.sub.3,
alkyl, or cycloalkyl. Preferred compounds according to formula XI
include:
##STR00022##
[0027] Yet another aspect of the present invention provides a
compound of formula XII, or a salt or prodrug thereof,
##STR00023##
wherein n is 0 to 4; Y and Z are taken together with the two
intervening carbon atoms to form a ring selected from phenyl and
thienyl, which ring is substituted at the C(7) position with at
least the substituent --C.ident.C--R, where R is H,
Si(CH.sub.3).sub.3, t-butyl, isopropyl, methyl, or cyclopropyl; Y'
and Z' are taken together with the two intervening carbon atoms to
form a ring selected from phenyl and thienyl, which ring is
substituted at the C(7)' position with at least the substituent
--C.ident.C--R', where R' is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; R.sub.1 and R.sub.1' are
independently one of H, CH.sub.3, CF.sub.3, CH.sub.2CF.sub.3,
CH.sub.2CH.sub.3, or cyclopropyl; and R.sub.2 and R.sub.2' are
independently a substituted or unsubstituted at least partially
unsaturated 5 or 6 membered cyclic or heterocyclic ring, wherein if
substituted the substituent is one or more of F, Cl, Br, or
NO.sub.2 at the 2'-position. Preferred compounds according to
formula XII include:
##STR00024##
[0028] A still further aspect of the present invention provides a
compound of the formula XIII, or a salt or prodrug thereof,
##STR00025##
wherein Y and Z are taken together with the two intervening carbon
atoms to form a ring selected from phenyl and thienyl, which ring
is substituted at the C(7) position with at least the substituent
--C.ident.C--R, where R is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; Y' and Z' are taken together
with the two intervening carbon atoms to form a ring selected from
phenyl and thienyl, which ring is substituted at the C(7)' position
with at least the substituent --C.ident.C--R', where R' is H,
Si(CH.sub.3).sub.3, t-butyl, isopropyl, methyl, or cyclopropyl;
R.sub.1 and R.sub.1' are independently one of H, CH.sub.3,
CF.sub.3, CH.sub.2CH.sub.3, CH.sub.2CF.sub.3, or cyclopropyl;
R.sub.2 and R.sub.2' are independently a substituted or
unsubstituted at least partially unsaturated 5 or 6 membered cyclic
or heterocyclic ring, wherein if substituted the substituent is one
or more of F, Cl, Br, or NO.sub.2 at the 2'-position; and B is O or
NH and wherein --BCH.sub.2B-- is optionally replaced with
--N(R.sub.7)--N(R.sub.7)--, where R.sub.7 is one of H, CH.sub.3,
alkyl, or cycloalkyl. Preferred compounds according to formula XIII
include:
##STR00026##
[0029] Yet another aspect of the present invention provides a
compound of the formula XIV, or a salt or prodrug thereof,
##STR00027##
wherein Y and Z are taken together with the two intervening carbon
atoms to form a ring selected from phenyl and thienyl, which ring
is substituted at the C(7) position with at least the substituent
--C.ident.C--R, where R is H, Si(CH.sub.3).sub.3, t-butyl,
isopropyl, methyl, or cyclopropyl; Y' and Z' are taken together
with the two intervening carbon atoms to form a ring selected from
phenyl and thienyl, which ring is substituted at the C(7)' position
with at least the substituent --C.ident.C--R', where R' is H,
Si(CH.sub.3).sub.3, t-butyl, isopropyl, methyl, or cyclopropyl;
R.sub.1 and R.sub.1' are independently one of H, CH.sub.3,
CF.sub.3, CH.sub.2CH.sub.3, CH.sub.2CF.sub.3, or cyclopropyl;
R.sub.2 and R.sub.2' are independently a substituted or
unsubstituted at least partially unsaturated 5 or 6 membered cyclic
or heterocyclic ring, wherein if substituted the substituent is one
or more of F, Cl, Br, or NO.sub.2 at the 2'-position; and B is O,
NH, or --N(R.sub.7)--N(R.sub.7)--, where R.sub.7 is one of H,
CH.sub.3, alkyl, or cycloalkyl. Preferred compounds according to
formula XIV include:
##STR00028##
[0030] Yet another aspect of the present invention provides a
compound of the formula XV, or a salt or prodrug thereof,
##STR00029##
wherein
[0031] n=1, n=2; R=H, SiMe.sub.3, tBu, CH.sub.3,
##STR00030##
Ar=phenyl, 2'-fluorophenyl, 2-thienyl, 3-thienyl, 2-pyridyl,
2-pyridyl N--O; X=N or CH
[0032] Still another compound (XVI) of the present invention
is:
##STR00031##
wherein
[0033] R=H, SiMe.sub.3, tBu, CH.sub.3,
##STR00032##
Ar=phenyl, 2'-fluorophenyl, 2-thienyl, 3-thienyl, 2-pyridyl,
2-pyridyl N--O; X=N or CH
[0034] Still another compound (XVII) of the present invention
is:
##STR00033##
wherein
[0035] R=H, SiMe.sub.3, tBu, CH.sub.3,
##STR00034##
Ar=phenyl, 2'-fluorophenyl, 2-thienyl, 3-thienyl, 2-pyridyl,
2-pyridyl N--O; Y=O, S, NHCH.sub.3
[0036] Another compound (XVIII) of the present invention is:
##STR00035##
wherein
[0037] n=0, n=1; R=H, SiMe.sub.3, tBu, CH.sub.3,
##STR00036##
Ar=phenyl, 2'-fluorophenyl, 2-thienyl, 3-thienyl, 2-pyridyl,
2-pyridyl N--O; Y=O, S, NHCH.sub.3
[0038] Yet another compound (XIX) of the present invention is:
##STR00037##
wherein
[0039] R=H, SiMe.sub.3, tBu, CH.sub.3,
##STR00038##
Ar=phenyl, 2'-fluorophenyl, 2-thienyl, 3-thienyl, 2-pyridyl,
2-pyridyl N--O; Y=O, S, NHCH.sub.3
[0040] Still another compound (XX) of the present invention is:
##STR00039##
wherein
[0041] R=H, SiMe.sub.3, tBu, CH.sub.3,
##STR00040##
Ar=phenyl, 2'-fluorophenyl, 2-thienyl, 3-thienyl, 2-pyridyl,
2-pyridyl N--O; Y=O, S, NHCH.sub.3
[0042] A further compound (XXI) of the present invention is:
##STR00041##
wherein
[0043] R=H, SiMe.sub.3, tBu, CH.sub.3,
##STR00042##
Ar=phenyl, 2'-fluorophenyl, 2-thienyl, 3-thienyl, 2-pyridyl,
2-pyridyl N--O; Y=O, S, NHCH.sub.3
[0044] Compounds (XV) to (XXI) above can also have R as CF.sub.3,
CCl.sub.3, or CBr.sub.3.
[0045] A still further aspect of the present invention provides
compositions comprising compounds of the above kind in a
pharmaceutically acceptable carrier. Such pharmaceutically
acceptable carriers are well known in the art.
[0046] Another aspect of the invention provides a method for the
treatment and/or prevention of anxiety which comprises
administering to a patient in need of such treatment an effective
amount of a compound of the above kinds, or a pharmaceutically
acceptable salt thereof or a prodrug thereof.
[0047] In the above embodiments by "alkyl" we mean a straight or
branched halogenated or unhalogenated alkyl group having 1-6 carbon
atoms. By "cycloalkyl" we mean one containing 3-7 carbon atoms.
Also, in the above embodiments by "cyclic" we prefer a phenyl group
and by "heterocyclic" we prefer a 2-pyridine or a 2- or
3-thiophene.
[0048] The compounds of the present invention are GABA.sub.A
receptor ligands which exhibit anxiolytic activity due to increased
agonist efficacy at GABA.sub.A/.alpha.2, GABA.sub.A/.alpha.3 and/or
GABA.sub.A/.alpha.5 receptors. The compounds in accordance with
this invention may possess at least 2-fold, suitably at least
5-fold, and advantageously at least a 10-fold, selective efficacy
for the GABA.sub.A/.alpha.2, GABA.sub.A/.alpha.3, and/or
GABA.sub.A/.alpha.5 receptors relative to the GABA.sub.A/.alpha.1
receptors. However, compounds which are not selective in terms of
their agonist efficacy for the GABA.sub.A/.alpha.2,
GABA.sub.A/.alpha.3, and/or GABA.sub.A/.alpha.5 receptors are also
encompassed within the scope of the present invention. Such
compounds will desirably exhibit functional selectivity by
demonstrating anxiolytic activity with decreased
sedative-hypnotic/muscle relaxant/ataxic activity due to decreased
efficacy at GABA.sub.A/.alpha.1 receptors.
[0049] For use in medicine, the salts of the compounds of formulas
(I)-(XXI) will be pharmaceutically acceptable salts. Other salts
may, however, be useful in the preparation of the compounds
according to the invention or of their pharmaceutically acceptable
salts. Suitable pharmaceutically acceptable salts of the compounds
of this invention include acid addition salts which may, for
example, be formed by mixing a solution of the compound according
to the invention with a solution of a pharmaceutically acceptable
acid such as hydrochloric acid, sulphuric acid, methanesulphonic
acid, fumaric acid, maleic acid, succinic acid, acetic acid,
benzoic acid, oxalic acid, citric acid, tartaric acid, carbonic
acid or phosphoric acid. Furthermore, where the compounds of the
invention carry an acidic moiety, suitable pharmaceutically
acceptable salts thereof may include alkali metal salts, e.g.
sodium or potassium salts, alkaline earth metal salts, e.g. calcium
or magnesium salts; and salts formed with suitable organic ligands,
e.g. quaternary ammonium salts.
[0050] The present invention includes within its scope prodrugs of
the compounds of formulas (I)-(XXI) above. In general, such
prodrugs will be functional derivatives of the compounds of
formulas (I)-(XXI) which are readily convertible in vivo into the
required compound of formulas (I)-(XXI). Conventional procedures
for the selection and preparation of suitable prodrug derivatives
are described, for example, in Design of Prodrugs, ed. H.
Bundgaard, Elsevier, 1985.
[0051] Where the compounds according to the invention have at least
one asymmetric center, they may accordingly exist as enantiomers.
Where the compounds according the invention possess two or more
asymmetric centers, they may additionally exist as
diastereoisomers. It is to be understood that all such isomers and
mixtures thereof in any proportion are encompassed within the scope
of the present invention.
[0052] The compounds according to the present invention exhibit
anxiolytic activity, as may be demonstrated in rats by a positive
response in a preclinical test for anti-anxiety efficacy (e.g.,
situational anxiety or defensive withdrawal). Moreover, the
compounds of the invention are substantially non-sedating and
non-ataxic as may be confirmed by an appropriate result obtained
from the locomotor activity test and rotorod paradigm,
respectively.
[0053] The compounds according to the present invention may also
exhibit anticonvulsant activity. This can be demonstrated by the
ability to block pentylenetetrazole-induced seizures in
rodents.
[0054] The invention also provides pharmaceutical compositions
comprising one or more compounds of this invention in association
with a pharmaceutically acceptable carrier. Preferably these
compositions are in unit dosage forms such as tablets, pills,
capsules, powders, granules, sterile parenteral solutions or
suspensions, metered aerosol or liquid sprays, drops, ampoules,
auto-injector devices or suppositories; for oral, parenteral,
intranasal, sublingual or rectal administration, or for
administration by inhalation or insufflation. It is also envisioned
that the compounds of the present invention may be incorporated
into transdermal patches designed to deliver the appropriate amount
of the drug in a continuous fashion. For preparing solid
compositions such as tablets, the principal active ingredient is
mixed with a pharmaceutical carrier, e.g. conventional tableting
ingredients such as corn starch, lactose, sucrose, sorbitol, talc,
stearic acid, magnesium stearate, dicalcium phosphate or gums, and
other pharmaceutical diluents, e.g. water, to form a solid
preformulation composition containing a homogeneous mixture for a
compound of the present invention, or a pharmaceutically acceptable
salt thereof. When referring to these preformulation compositions
as homogeneous, it is meant that the active ingredient is dispersed
evenly throughout the composition so that the composition may be
easily subdivided into equally effective unit dosage forms such as
tablets, pills and capsules. This solid performulation composition
is then subdivided into unit dosage forms of the type described
above containing from 0.1 to about 500 mg of the active ingredient
of the present invention. Typical unit dosage forms contain from 1
to 100 mg, for example, 1, 2, 5, 10, 25, 50 or 100 mg, of the
active ingredient. The tablets or pills of the novel composition
can be coated or otherwise compounded to provide a dosage from
affording the advantage of prolonged action. For example, the
tablet or pill can comprise an inner dosage and an outer dosage
component, the latter being in the form of an envelope over the
former. The two components can be separated by an enteric layer
which, serves to resist disintegration in the stomach and permits
the inner component to pass intact into the duodenum or to be
delayed in release. A variety of materials can be used for such
enteric layers or coatings, such materials including a number of
polymeric acids and mixtures of polymeric acids with such materials
as shellac, cetyl alcohol and cellulose acetate.
[0055] The liquid forms in which the novel compositions of the
present invention may be incorporated for administration orally or
by injection include aqueous solutions, suitably flavored syrups,
aqueous or oil suspensions, and flavored emulsions with edible oils
such as cottonseed oil, sesame oil, coconut oil or peanut oil, as
well as elixirs and similar pharmaceutical vehicles. Suitable
dispersing or suspending agents for aqueous suspensions include
synthetic and natural gums such as tragacanth, acacia, alginate,
dextran, sodium caboxymethylcellulose, methylcellulose,
polyvinylpyrrolidone or gelatin.
[0056] In the treatment of anxiety, suitable dosage level is about
0.01 to 250 mg/kg per day, preferably about 0.05 to 100 mg/kg per
day, and especially about 0.05 to 5 mg/kg per day. The compounds
may be administered on a regimen of 1 to 4 times per day, or on a
continuous basis via, for example, the use of a transdermal
patch.
DETAILED DESCRIPTION OF THE INVENTION
##STR00043##
[0058] The bromide 1 available from reference .sup.1 was reacted
with trimethylsilyacetylene in the presence of a palladium catalyst
to provide trimethylsilyl analog 2..sup.4,5,6 This product was
methylated with methyl iodide/sodium hydride to give the N-methyl
benzodiazepine 3. This was subjected to fluoride-mediated
desilation to furnish 4 (QHII-066).
Procedure for QHII-066
[0059]
7-Trimethylsilylacetyleno-5-phenyl-1,3-dihydrobenzo[e]-1,4-diazepin-
-2-one 2..sup.4,5,8 A mixture of 1.sup.1 (1 g, 3.17 mmole available
from reference 1) in triethyl amine (30 mL) and CH.sub.3CN (20 mL)
with trimethylsilylacetylene (622.7 mg, 6.34 mmole) and
bis(tri-phenylphosphine)-palladium (II) acetate (118 mg, 0.16 mmol)
was heated to reflux under nitrogen. After 12 hours, the reaction
mixture was cooled to room temperature and filtered. The filtrate
was concentrated in vacuum and the residue was treated with a
saturated aqueous solution of NaHCO.sub.3 (30 mL), and extracted
with CH.sub.2Cl.sub.2 (3.times.50 mL). The organic layers were
combined and washed with brine and dried (Na.sub.2SO.sub.4). After
removal of solvent under reduced pressure, the residue was purified
via flash chromatography (silica gel, EtOAc/hexanes:1/1) to furnish
3 as a yellow powder (791 mg, 75%): mp: 190-191.5.degree. C.; IR
(KBr) 3011, 2281, 1686, 1610, 1486, 1325, 1249, 839, 700 cm.sup.-1;
.sup.1H NMR (CDCl.sub.3) .delta. 0.21 (s, 9H), 4.31 (s, 2H), 7.09
(d, 1H, J=8.25 Hz), 7.21-7.61 (br, 7H), 10.17 (s, 1H); MS (CI) m/e
(relative intensity) 333 (M.sup.++1, 100). This material was used
in the next step.
[0060]
1-Methyl-7-trimethylsilylacetyleno-5-phenyl-1,3-dihydrobenzo[e]-1,4-
-diazepin-2-one 3..sup.7 A mixture of 2 (485 mg, 1.46 mmol) was
dissolved in dry THF (20 mL) at 0.degree. C. and NaH (60% in
mineral oil, 70 mg, 1.75 mmol) was added to the solution in one
portion. The slurry was then stirred for 20 min at 0.degree. C. and
CH.sub.3I (311 mg, 2.19 mmol) was added to the mixture and it was
warmed up to room temperature. After the mixture stirred for 3
hours at room temperature, the THF was then removed under reduced
pressure. The residue was purified by flash chromatography
[hexanes/EtOAc (1:4)] to provide the title compound 3 (303 mg, 60%)
as a white solid: mp: 177-178.degree. C.; IR (KBr) 2954, 2147,
1687, 1612, 1491, 1382, 1115, 1075, 839, 700 cm.sup.-1; .sup.1HNMR
(CDCl.sub.3) 8 (ppm), 3.18 (s, 3H), 3.54 (d, 1H, J=10.8 Hz), 4.60
(d, 1H. J=10.8 Hz), 7.05 (s, 1H), 7.07 (d, 1H, J=8.58 Hz),
7.20-7.27 (m, 3H), 7.37-7.42 (m, 3H); MS (EI) m/e 346 (M.sup.+,
90), 318 (100), 303(19), 165(22), 151 (20). Anal. Calcd. for
C.sub.21H.sub.22N.sub.2OSi: C, 72.79; H, 6.40; N, 8.08. Found: C,
72.50; H, 6.68; N, 8.04.
[0061]
1-Methyl-7-acetyleno-5-phenyl-1,3-dihydro-benzo[e]-1,4-diazepin-2-o-
ne 4 (QHII-066)..sup.7 A solution of 3 (100 mg) in THF (30 mL) was
treated with tetrabutylammonium fluoride (1M in THF). The mixture
was stirred for 20 minutes at room temperature before water (30 mL)
was added. The mixture was then extracted with EtOAc (3.times.30
mL). The combined organic extracts were washed with brine and dried
(Na.sub.2SO.sub.4). The solvent was removed under vacuum and the
residue which resulted was passed through a wash column (silica
gel, EtOAc/hexanes: 4/1) to give 4 (QHII-066) as light yellow
crystals (71 mg, 90%): mp: 163-165.degree. C.; IR (KBr) 2965, 1680,
1605, 1387, 1121, 833, 747 cm.sup.-1; 1HNMR (CDCl.sub.3) .delta.
(ppm) 3.38 (s, 3H), 3.75 (d, 1H, J=10.8 Hz), 4.80 (d, 1H, J=10.9
Hz), 5.28 (s, 1H), 7.29 (d, 1H, J=8.5 Hz), 7.35-7.45 (m, 4H),
7.55-7.59 (m, 2H), 7.62 (dd, 1H, J=8.5 Hz, 2.0 Hz); MS (EI) m/e
(relative intensity) 274 (M.sup.+, 100), 259 (12), 246 (100), 189
(12).122(19), 105 (42). Anal. Calcd. for
C.sub.18H.sub.14N.sub.2O.2/3H.sub.2O, Calculated: C, 75.51; H,
4.89; N, 9.78. Found: C, 75.59; H, 5.17; N, 9.62.
##STR00044##
[0062] The bromide 1 was reacted with diethylphosphorochloridate in
the presence of sodium hydride, followed by addition of ethyl
isocyanoacetate to provide the ester 5. This was converted to the
trimethylsilylacetyleno compound 6 (XLiXHeII-048) under standard
conditions (Pd-mediated, Heck-type coupling)..sup.8 Treatment of 6
with fluoride gave the title compound 7 (XHeII-053).
Procedure for XHe-II-053
[0063] Ethyl
8-bromo-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxylate
5. This benzodiazepine 5 was obtained in 45% yield from 1.sup.1
analogous to the literature procedure.sup.2 as a white solid. 2:
mp: 174-175.degree. C.; IR (KBr) 2978, 1712, 1609, 1491 cm.sup.-1;
.sup.1H NMR (CDCl.sub.3) .delta. 1.44 (t, 3H, J=7.1 Hz), 4.09 (d,
1H, J=12.1 Hz), 4.38-4.49 (m, 2H), 6.08 (d, 1H, J=12.3 Hz),
7.40-7.53 (m, 6H), 7.60 (d, 1H, J=2.2 Hz), 7.82 (dd, 1H, J=8.6 Hz
and 2.2 Hz), 7.95 (s, 1H); MS (EI) m/e (relative intensity) 411
(34), 410 (M.sup.+, 8), 409 (34), 365 (61), 363 (61), 337 (100),
335 (100), 285 (21), 232, (17). Anal. Calcd. for
C.sub.20H.sub.16BrN.sub.3O.sub.2: C, 58.55; H, 3.93; N, 10.24.
Found: C, 58.30, H, 3.91; N, 9.90.
[0064] Ethyl
8-trimethylsilylacetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepi-
ne-3-carboxylate 6 (XLiXHeII-048)..sup.4,5,8 A mixture of bromide 5
(0.3 g, 0.73 mmol), trimethylsilylacetylene (0.143 g, 1.46 mmol)
and bis(triphenylphosphine)-palladium-(II) acetate (55 mg, 0.073
mmol) in a mixed solvent system of toluene (20 mL) and anhydrous
TEA (50 mL) was heated to reflux under argon. After stirring for 12
hours at reflux, the mixture was cooled to room temperature and the
precipitate which formed was removed by filtration. The filtrate
was concentrated under reduced pressure and the residue was treated
with a saturated aqueous solution of NaHCO.sub.3 (20 mL), and
extracted with CHCl.sub.3 (3.times.25 mL). The combined extracts
were washed with brine and dried (Na.sub.2SO.sub.4). After removal
of solvent under reduced pressure, the residue was purified by
flash chromatography (silica gel, EtOAc) to afford 6 (XLiXHeII-048)
as a white solid (0.29 g, 93%). This benzodiazepine can also be
obtained from 2 in 45% yield by following the same procedure 6
(XLiXHeII-048): mp: 170-172.degree. C.; IR (KBr) 2958, 2152, 1718
cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta. 0.23 (s, 9H), 1.42 (t,
3H, J=7.2 Hz), 4.04 (d, 1H, J=12.6 Hz), 4.41 (m, 2H, J=7.2 Hz),
6.23 (d, 1H, J=12.6 Hz), 7.35-7.55 (m, 7H), 7.73 (dd, 1H, J=8.3 Hz,
J=1.9 Hz), 7.93 (s, 1H); MS (EI) m/e (relative intensity) 427
(M.sup.+, 76), 412 (5), 381 (55), 353 (100) 303 (10), 287 (7).
Anal. Calcd. for C.sub.25H.sub.25N.sub.3O.sub.2Si.1/3 EtOAc: C,
69.22; H, 6.01; N, 9.20. Found: C, 68.87; H, 5.81; N, 9.37.
[0065] Ethyl
8-acetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxyla-
te 7 (XHeII-053)..sup.7 A solution of 6 (XLiXHeII-048) (0.17 g,
0.41 mmol), in THF (15 mL) was treated with Bu.sub.4NF.H.sub.2O
(0.16 g, 0.62 mmol). The mixture which resulted was allowed to stir
for 30 min at room temperature after which the mixture was added to
H.sub.2O (10 mL) and extracted with EtOAc (3.times.25 mL). The
combined organic extracts were washed with brine (25 mL) and dried
(Na.sub.2SO.sub.4). After removal of solvent under reduced
pressure, the residue was purified by a wash column (silica gel,
EtOAc) to furnish 7 (XHeII-053) (0.12 g, 85%) as a white solid: mp
237-239.degree. C.; IR (KBr) 3159, 3107, 2092, 1721, 1606
cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta. 1.44 (t, 3H, J=7.1 Hz),
3.20 (s, 1H), 4.13 (d, 1H, J=10.22 Hz), 4.41-4.48 (m, 2H), 6.11 (d,
1H, J=12 Hz), 7.42-7.63 (m, 7H), 7.81 (dd, 1H, J=8.3 Hz and 1.8
Hz), 8.03 (s, 1H); MS (EI) m/e (relative intensity) 355 (M.sup.+,
83), 309 (70), 281 (100), 253 (12), 231 (18), 178 (20). Anal.
Calcd. for C.sub.22H.sub.17N.sub.3O.sub.2.3/4H.sub.2O: C, 71.63; H,
5.05; N, 11.39. Found: C, 71.27; H, 4.71; N, 11.03.
##STR00045##
[0066] The bromide 1, available from reference 1, was stirred with
the di-4-morpholino-phosphinic chloride, followed by addition of
acetylhydrazide to furnish triazolo-benzodiazepine 8. This material
8 was subjected to a Heck-type coupling reaction (TMS-C.ident.CH,
Pd-mediated).sup.4,7,8 to furnish ligand 9. This analog was
converted into 10 (XLi270) on stirring with fluoride anion as shown
in Scheme 3.
Procedure for XLi 270
[0067]
8-Bromo-1-methyl-6-phenyl-4H-s-triazolo[4,3-a][1,4]benzodiazepine
8..sup.3 A solution of 1.sup.1 (1 g, 3.07 mmol of
7-bromo-5-phenyl-1,4-benzodiazepine-2-one) in dry THF (20 mL) was
cooled in an ice-water bath and a 60% dispersion of sodium hydride
(152.2 mg) was added in one portion. After 20 minutes,
di-4-morpholinylphosphinic chloride.sup.3 (943.9 mg, 4.76 mmol) was
added at 0.degree. C. and this was stirred for 30 minutes and
allowed to warm to room temperature. The mixture was stirred for
1.5 hours. To this mixture was then added a solution of
acetylhydrazide (521.9 mg, 7.14 mmol) in dry butanol (5 mL) and
stirring was continued at room temperature for 10 min. The solvents
were evaporated and the residue was dissolved in butanol (10 mL)
and heated to reflux for 5 hours. Butanol was removed under reduced
pressure and the residue was partitioned between CH.sub.2Cl.sub.2
(50 mL) and water (50 mL). The water layer was extracted by
CH.sub.2Cl.sub.2 (3.times.30 mL). The combined organic layer was
washed by brine (30 mL). The organic layer was dried
(Na.sub.2SO.sub.4) and the solvent was removed under vacuum. The
residue was purified by flash chromatography (silica gel) to
provide pure 8 [539.5 mg (40% yield)] as a white solid: mp
268.5-270.degree. C.; IR (KBr) 2358, 1607, 1538, 1484, 1311, 1000,
801, 697 cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta. 2.82 (s, 3H),
4.11 (d, 1H, J=12.8 Hz), 5.49 (d, 1H, J=12.8 Hz), 7.21-7.68 (m,
7H), 7.75 (dd, 1H, J=0.58 Hz, J=1.5 Hz); MS (EI) m/e (relative
intensity) 354 (34), (M.sup.+, 16), 352 (34), 325(33), 323 (34),
273 (63), 245 (31), 232 (19), 204 (100), 183(23), 177 (36), 151
(24). Anal. Calcd. for C.sub.17H.sub.13BrN.sub.4: C, 57.81; H,
3.71; N, 15.86. Found C, 57.57; H, 3.64: N, 15.70.
[0068]
8-Trimethylsilylacetylenyl-1-methyl-6-phenyl-4H-s-triazolo[4,3-a][1-
,4]-benzodiazepine (XLi269). A mixture of 8
(8-bromo-1-methyl-6-phenyl-4-H-s-triazolo-[4,3-a][1,4]benzodiazepine,
300 mg, 0.85 mmol), trimethylsilylacetylene (208.5 mg, 2.12 mmol)
and bis(triphenylphosphine)-palladium(II) acetate in a mixed
solvent system of EtN.sub.3 (5 mL) and CH.sub.3CN (8 mL) was heated
to reflux under nitrogen. After stirring for 6 hours at reflux. The
mixture was cooled to room temperature. The mixture was
concentrated under reduced pressure and H.sub.2O (30 mL) was added.
The mixture was extracted with CH.sub.2Cl.sub.2 (3.times.50 mL).
The combined extracts were washed with brine and dried
(Na.sub.2SO.sub.4). After removal of solvent under reduced
pressure, the residue was purified by flash chromatography (silica
gel, EtOH/EtOAc) to afford benzodiazepine 9 (185 mg, 60% yield) as
a white solid: mp 229-233.degree. C.; IR (KBr) 2957, 2156, 1609,
1537, 1491, 1424, 1315, 1249, 881, 844, 750 cm.sup.-1; .sup.1H NMR
(CDCl.sub.3) .delta. 0.23 (s, 9H), 2.68 (s, 3H), 4.11 (d, 1H,
J=12.5 Hz), 5.49 (d, 1H, J=13.0 Hz), 7.21-7.68 (m, 7H), 7.75 (dd,
1H, J=8.5 Hz, J=1.5 Hz); MS (EI) m/e (relative intensity) 370
(M.sup.+, 80), 355 (44), 341 (60), 286 (34), 177 (51), 163 (52) 143
(100), 129 (19), 115 (28). Anal. Calcd. for
C.sub.22H.sub.22N.sub.4Si: C, 71.31; H, 5.98; N, 15.12. Found: C,
70.90; H, 5.93; N, 15.08.
[0069]
8-Acetylenyl-1-methyl-6-phenyl-4H-s-triazolo[4,3-a][1,4]benzodiazep-
ine 10 (Xli-270)..sup.7A solution of 9
[trimethylsilylacetylenyl-1-methyl-6-phenyl-4H-s-triazolo-[4,3-a]-[1,4]-b-
enzodiazepine (106.4 mg, 0.288 mmol)] in dry THF (20 mL) was
treated with Bu.sub.4NF (1.0 M in THF, 112.8 mg, 0.431 mmol). The
mixture which resulted was allowed to stir for 5 min at room
temperature after which the mixture was added to H.sub.2O (10 mL)
and extracted with CH.sub.2Cl.sub.2 (3.times.25 mL). The combined
organic extracts were washed with brine (25 mL) and dried
(Na.sub.2SO.sub.4). After removal of solvent under reduced
pressure, the residue was crystallized from EtOAc to provide
benzodiazepine 10 (XLi270) (66.8 mg, 80% yield) as a white solid:
mp>250.degree. C. (dec); IR (KBr) 3198, 2158, 1609, 1538, 1491,
1425, 1317, 1002, 838, 748, 695 cm.sup.-1; .sup.1H NMR (CDCl.sub.3)
.delta. 2.78 (s, 3H), 3.15 (s, 1H), 4.11 (d, 2H, J=12.8 Hz), 5.91
(d, 1H, J=12.8 Hz), 7.35-7.85 (m, 8H); MS (EI) (relative intensity)
298 (M.sup.+, 100), 269 (78), 230 (48), 228 (65), 201 (20), 127
(65), 115 (42), 101 (54). Anal. Calcd. for
C.sub.19H.sub.14N.sub.4.1/2 CH.sub.3OH: C, 74.50; H, 5.13; N,
17.82. Found: C, 74.33; H, 4.83; N, 17.77.
##STR00046##
[0070] The 7-bromo-2'-fluorobenzodiazepine 12 (available from
reference 1) was reacted with sodium hydride and
diethylphosphorochloridate and this was followed by addition of
ethyl isocyanoacetate to provide benzimidazo intermediate 13
(JYI-032),.sup.2 as illustrated in Scheme 4. This material was
heated with trimethysilylacetylene in a Heck-type coupling
reaction.sup.8 to provide the trimethylsilyl analog 14 (JYI-038).
The silyl group was removed from 14 on treatment with fluoride
anion to furnish 15, a 2'-fluoro analog of XHeII-053, in excellent
yield.
Procedure:
[0071] Ethyl
8-bromo-6-(2'-fluorophenyl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-car-
boxylate 13 (JYI-032). A solution of 12.sup.1 (7.0 g, 21.0 mmol) in
THF (50 mL) was cooled in ice-water, and sodium hydride (1.0 g,
25.2 mmol) was added in one portion. After 30 min, diethyl
phosphorochloridate (5.62 g, 31.5 mmol) was added dropwise, and the
solution which resulted was stirred continuously for 30 min with
cooling from an ice bath. A solution of ethyl isocyanoacetate (4.22
g, 25.2 mmol) and sodium hydride (1.17 g, 29.4 mmol) in THF (10
mL), which had stirred for 30 min with ice-bath cooling, was added
slowly via a cannula. After stirring for another 30 min with
cooling, the reaction mixture was allowed to stir at room
temperature overnight. The mixture was then added to H.sub.2O (10
mL) and extracted with EtOAc (3.times.50 mL). The combined organic
extracts were washed with brine (2.times.50 mL) and dried
(Na.sub.2SO.sub.4). The solvent was evaporated under reduced
pressure and the residue was purified by flash chromatography
(silica gel, hexanes/EtOAc: 2/1) to afford 13 (JYI-032, 5.2 g, 58%)
as a white solid: mp 200-201.5.degree. C.; IR (KBr) 2977, 1718,
1610, 1491, 1450 cm.sup.-1; .sup.1H NMR (DMSO-d.sub.6) .delta. 1.30
(t, 3H, J=4.2 Hz), 4.28 (bs, 1H), 4.30 (q, 2H, J=4.2 Hz), 5.75 (bs,
1H), 7.20 (t, 1H, J=5.6 Hz), 7.30 (t, 1H, J=4.5 Hz), 7.40 (s, 1H),
7.54 (m, 2H), 7.85 (d, 1H, J=5.2 Hz), 7.96 (dd, 1H, J=5.2 Hz and
1.3 Hz), 8.44 (s, 1H); MS (EI) m/e (relative intensity) 428 (7),
381 (58), 355 (100), 303 (37), 274 (36), 247 (35), 234 (52), 154
(71), 127 (62). Anal Calcd. for C.sub.20H.sub.15N.sub.3O.sub.2FBr:
C, 56.09; H, 3.53; N, 9.81. Found: C, 56.02; H, 3.51; N, 9.58.
[0072] Ethyl
8-trimethylsilylacetylenyl-6-(2'-fluorophenyl)-4H-benzo[f]-imidazo[1,5-a]-
[1,4]diazepine-3-carboxylate 14 (JYI-038). A mixture of bromide 13
(JYI-032, 1.40 g, 3.3 mmol), trimethylsilylacetylene (0.65 g, 6.6
mmol) and bis(triphenylphosphine)-palladium (II) acetate (0.25 g,
0.33 mmol) in a mixed solvent system of CH.sub.3CN (80 mL) and
anhydrous triethylamine (50 mL) was heated to reflux under argon.
After stirring for 2 h at reflux, the mixture was cooled to room
temperature and the precipitate which formed was removed by
filtration. The filtrate was concentrated under reduced pressure
and the residue was treated with a saturated aqueous solution of
NaHCO.sub.3 (40 mL), and extracted with CHCl.sub.3 (3.times.50 mL).
The combined organic extracts were washed with brine (2.times.20
mL) and dried (Na.sub.2SO.sub.4). After removal of solvent under
reduced pressure, the residue was purified by flash chromatography
(silica gel, hexanes/EtOAc: 3/1) to afford 14 (JYI-038, 1.2 g, 82%)
as a white solid: mp 196-197.5.degree. C.; IR (KBr) 2959, 2157,
1709, 1613, 1494, 1451, 1252 cm.sup.-1; .sup.1H NMR (DMSO-d.sub.6)
.delta. 0.20 (s, 9H), 1.32 (t, 3H, J=7.1 Hz), 4.18 (bs, 1H), 4.32
(q, 2H, J=7.1 Hz), 5.78 (bs, 1H), 7.25 (t, 1H, J=11.5 Hz),
7.30-7.35 (m, 4H), 7.81 (d, 1H, J=6.6 Hz), 7.93 (d, 1H, J=8.4 Hz),
8.49 (s, 1H); MS (EI) m/e (relative intensity) 445 (37), 399 (51),
371 (100), 235 (71), 192 (66), 178 (75). Anal. Calcd. for
C.sub.25H.sub.24N.sub.3O.sub.2FSi: C, 67.39; H, 5.42; N, 9.43.
Found: C, 66.98; H, 5.46; N, 9.19.
[0073]
8-Acetyleno-6-(2'-fluorophenyl)-4H-benzo[f]imidazo[1,5-a][1,4]diaze-
pine-3-carboxylate 15 (JY-XHE-053). A solution of 14 (JYI-038, 80
mg, 0.18 mmol) in THF (5 mL) was treated with Bu.sub.4NF (0.5 mL,
1.0M solution in THF). The mixture which resulted was allowed to
stir for 5 min at room temperature after which the mixture was
added to H.sub.2O (5 mL) and extracted with EtOAc (3.times.10 mL).
The combined organic extracts were washed with brine (2.times.10
mL) and dried (Na.sub.2SO.sub.4). The solvent was removed under
reduced pressure and the residue was purified by flash
chromatography (silica gel, EtOAc) to afford 15 (JY-XHE-053, 67 mg,
80%) as a white solid: mp 223.5-224.5.degree. C.; IR (KBr) 3288,
2979, 1712, 1621, 1491, 1255, 1190 cm.sup.-1; .sup.1H NMR
(DMSO-d.sub.6) .delta. 1.34 (t, 3H, J=7.1 Hz), 4.27 (bs, 1H), 4.36
(q, 2H, J=7.1 Hz), 4.47 (s, 1H), 5.80 (bs, 1H), 7.22 (t, 1H, J=8.4
Hz), 7.30-7.60 (m, 4H), 7.85 (d, 1H, J=6.6 Hz), 7.92 (d, 1H, J=8.4
Hz), 8.83 (s, 1H); MS (EI) m/e (relative intensity) 373 (28), 327
(47), 299 (100), 249(22), 178 (50). Anal. Calcd. for
C.sub.22H.sub.16N.sub.3O.sub.2F.1/2H.sub.2O: C, 69.10; H, 4.48; N,
10.99. Found: C, 69.19; H, 4.39; N, 10.68.
##STR00047##
[0074] The 7-bromo-2'-fluorobenzodiazepine 12 was stirred with
sodium hydride and di-4-morpholinylphosphinic chloride, followed by
addition of acetic hydrazide, according to the published
procedure.sup.3 to provide triazolobenzodiazepine 16 (WI-73), as
illustrated in Scheme 5. This compound 16 underwent the
palladium-mediated Heck-type coupling reaction.sup.8 with
trimethylsilylacetylene to furnish the 8-trimethylsilyl substituted
analog 17 (JYI-72). Removal of the silyl group from 17 furnished
the 8-acetyleno triazolobenzodiazepine 18 (JYI-70).
Procedure:
[0075]
8-Bromo-1-methyl-6-(2'-fluorophenyl)-4H-s-triazolo[4,3-a][1,4]benzo-
diazepine 16 (JYI-73). A solution of 12 (JYI-032, 7.0 g, 21.0 mmol)
in THF (50 mL) was cooled in ice-water, and sodium hydride (0.72 g,
18 mmol) was added in one portion. After 1 hour,
di-4-morpholinylphosphinic chloride (4.84 g, 22.5 mmol) was added,
and the solution which resulted was stirred continuously for 2
hours at room temperature. To this mixture was then added a
solution of acetic hydrazide (2.47 g, 30 mmol) in n-BuOH (20 mL)
and stirring was continued at room temperature for 15 min. The
solvents were evaporated and the residue was dissolved in n-BuOH
(25 mL) and heated to reflux for 2 hours. n-Butanol was evaporated
and the residue was partitioned between CH.sub.2Cl.sub.2 and brine.
The CH.sub.2Cl.sub.2 layer was dried and removed under reduced
pressure after which the residue was purified by flash
chromatography (silica gel, EtOAc) to afford 16 (JYI-73, 2.2 g,
40%) as a white solid: mp 213-214.degree. C.; IR (KBr) 1610, 1484,
1426, 1314 cm.sup.-1; .sup.1H NMR (DMSO-d.sub.6) .delta. 2.56 (s,
3H), 4.28 (d, 1H, J=12.9 Hz), 5.26 (d, 1H, J=12.9 Hz), 7.24 (t, 1H,
J=8.3 Hz), 7.29 (t, 1H, J=7.2 Hz), 7.35 (s, 1H), 7.43-7.60 (m, 2H),
7.83 (d, 1H, J=8.7 Hz), 7.98 (dd, 1H, J=8.7 Hz and 2.3 Hz); MS (EI)
m/e (relative intensity) 371 (5), 341 (34), 222 (100), 195 (19),
181 (28), 111 (72). Anal. Calcd. for C.sub.17H.sub.12N.sub.4FBr: C,
55.01; H, 3.26; N, 15.09. Found: C, 54.76; H, 3.29; N, 14.74.
[0076]
8-Trimethylsilylacetylenyl-1-methyl-6-(2'-fluorophenyl)-4H-s-triazo-
lo[4,3-a][1,4]-benzodiazepine 17 (JYI-72). A mixture of bromide 16
(JYI-73, 1.40 g, 3.8 mmol), trimethylsilylacetylene (0.65 g, 6.6
mmol) and bis(triphenylphosphine)palladium (II) acetate (0.25 g,
0.33 mmol) in a mixed solvent system of CH.sub.3CN (80 mL) and
anhydrous triethylamine (50 mL) was heated to reflux under argon.
After stirring for 2 hours at reflux, the mixture was cooled to
room temperature and the precipitate which formed was removed by
filtration. The filtrate was concentrated under reduced pressure
and the residue was treated with a saturated aqueous solution of
NaHCO.sub.3 (40 mL), and extracted with CHCl.sub.3 (3.times.50 mL).
The combined organic extracts were washed with brine (2.times.10
mL) and dried (Na.sub.2SO.sub.4). After removal of solvent under
reduced pressure, the residue was purified by flash chromatography
(silica gel, EtOAc) to afford 17 (JYI-72, 1.15 g, 77%) as a gray
solid: mp 218-219.degree. C.; IR (KBr) 2958, 2157, 1612, 1537,
1493, 1452, 1317, 1249 cm.sup.-1; .sup.1H NMR (DMSO-d.sub.6)
.delta. 0.21 (s, 9H), 2.56 (s, 3H), 4.23 (s, 1H, J=12.9 Hz), 7.26
(t, 1H, J=8.4 Hz), 7.29-7.83 (m, 6H); MS (EI) m/e (relative
intensity) 388 (65), 373 (14), 359 (77), 304 (44), 152 (100). Anal.
Calcd. for C.sub.22H.sub.21N.sub.4SiF. 0.7H.sub.2O: C, 65.87; H,
5.62; N, 13.94. Found: C, 65.88; H, 5.34; N, 13.94.
[0077]
8-Acetyleno-1-methyl-6-(2'-fluorophenyl)-4H-s-triazolo[4,3-a][1,4]b-
enzodiazepine 18 (JYI-70). A solution of 17 (JYI-72, 2.0 g, 5 mmol)
in THF (20 mL) was treated with Bu.sub.4NF (4 mL, 1.0M solution in
THF). The mixture which resulted was allowed to stir for 5 min at
room temperature after which the mixture was added to H.sub.2O (20
mL) and extracted with CH.sub.2Cl.sub.2 (3.times.50 mL). The
combined organic extracts were washed with brine (2.times.15 mL)
and dried (Na.sub.2SO.sub.4). After removal of solvent under
reduced pressure, the residue was purified by flash chromatography
(silica gel, EtOAc/MeOH: 100/1) to afford 18 (JYI-70, 1.1 g, 70%)
as a pale yellow solid: mp>250.degree. C. (dec); IR (KBr) 3205,
1612, 1493, 1426, 1317 cm.sup.1; .sup.1H NMR (DMSO-d.sub.6) .delta.
2.54 (s, 3H), 4.22 (d, 1H, J=12.9 Hz), 4.39 (s, 1H), 5.26 (d, 1H,
J=12.9 Hz), 7.22 (t, 1H, J=8.3 Hz), 7.32-7.55 (m, 4H), 7.97 (m,
2H); MS (EI) m/e (relative intensity) 316 (72), 287 (100), 246
(69), 153 (16), 127 (62). Anal. Calcd. for
C.sub.19H.sub.13N.sub.4F.0.6 CH.sub.3OH: C, 70.16; H, 4.37; N,
16.55. Found: C, 69.98; H, 4.31; N, 16.70.
##STR00048## ##STR00049##
[0078] 2-Amino-5-bromo-2'-chlorobenzophenone 19 was obtained from
simple starting materials, 4-bromoaniline and 2-chlorobenzoyl
chloride, according to the improved conditions in the
literature..sup.9 The benzodiazepine 20, available from reference
1, was stirred with sodium hydride and di-4-morpholinophosphinic
chloride, followed by addition of acetylhydrazide to furnish
triazolobenzodiazepine 21 (dm-II-90)..sup.3 The ligand 22
(XLi-JY-DMH-TMS) was obtained by a Heck coupling reaction of 21
(dm-II-90) with trimethylsilylacetylene..sup.4,7,8 This compound
was converted into acetylene 23 (XLi-JY-DMH.sup.+).sup.7 on
stirring with fluoride anion as shown in Scheme 6.
[0079] 2-Amino-5-bromo-2'-chlorobenzophenone 19..sup.9
2-Chlorobenzoyl chloride (177 mL, 1.4 mol) was cooled in a 2-L
flask equipped with a condenser and a thermometer to 0.degree. C.
with an ice-water bath and 4-bromoaniline (100 g, 0.58 mol) was
added to the cooled solution. The mixture was heated to 120.degree.
C. and kept at this temperature for 1 h until analysis by TLC
indicated 4-bromoaniline had been consumed (EtOAc:hexane, 1:4). The
solution was heated to 160.degree. C. and anhydrous ZnCl.sub.2 (95
g, 0.70 mol, flamed dried) was added in one portion. The
temperature was increased to 195.degree. C. and stirring was
maintained at this temperature for 3 hr until no more bubbles were
evolved. The mixture was cooled to 120.degree. C. and aq HCl (12%,
350 mL) was added dropwise slowly. The mixture was kept at reflux
for 20 min, after which the aq layer was poured off. This procedure
with aq HCl was repeated 4 times. Water (350 mL) was then added,
and the mixture held at reflux for 20 min and then the water was
poured off. This was repeated several times until the solid was not
a block any more. Then H.sub.2SO.sub.4 (72%, 700 mL) was added to
the residue and the mixture was heated to reflux for about 1 hr
until the reaction mixture became a homogeneous dark colored
solution. The hot acidic solution was poured into a mixture of ice
and water with stirring. The precipitate which resulted was
filtered and washed with a large amount of cold water until the pH
value of the solid was about 6. The solid was then suspended in ice
water and aq NaOH (40%, 290 mL) was added carefully. The mixture
which resulted was stirred for 2 hrs. The solid was filtered and
washed with ice water. The suspension of the solid in ice water was
adjusted carefully to approximately pH=3 with aq H.sub.2SO.sub.4
(40%) dropwise. The solid which remained was filtered and washed
with water to neutrality. The yellow solid 19 (66.1 g, 37.0%) was
dried and used directly in the next step without further
purification. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 6.49 (s,
br, 2H), 6.65 (d, 1H, J=8.82 Hz), 7.26-7.8 (m, 6H).
8-Bromo-5-(2'-chlorophenyl)-1-methyl-4H-s-triazolo[4,3-a]-1,4-benzodiazepi-
ne 21 (dm-II-90)..sup.3
[0080] A solution of benzodiazepine 20 (20 g, 57 mmol, available
from reference 1) in dry THF (250 mL) was cooled to -5.degree. C.
and a 60% dispersion of sodium hydride (3.66 g, 92 mmol) was added
in one portion. The mixture was allowed to warm to rt with stirring
and the stirring was continued at rt until no more bubbles were
evolved. The suspension was cooled to -5.degree. C. after which
di-4-morpholinylphosphinic chloride (21.8 g, 86 mmol) was added and
this mixture was stirred for 30 min and allowed to warm to rt. The
mixture was stirred for an additional 1.5 hr. To the mixture was
then added a solution of acetylhydrazide (9.42 g, 114 mmol) in
butanol (60 mL) and stirring was continued at rt for 10 min. The
solvent was removed under reduced pressure and the residue was
taken up in butanol (100 mL) and held at reflux for 2 hr. Butanol
was removed under reduced pressure and the residue was partitioned
between CH.sub.2Cl.sub.2 (200 mL) and H.sub.2O (100 mL). The aq
layer was extracted 4 times and the organic layers combined. The
organic layer was washed with brine and dried (Na.sub.2SO.sub.4).
After the solvent was removed under reduced pressure, the residue
was crystallized from EtOAc-Et.sub.2O to provide the pure
triazolobenzodiazepine 21 (dm-II-90, 14 g, 63.2%) as a yellow
solid: mp 265-267.degree. C. [lit 274-275.degree. C.].sup.(10); IR
(KBr) 3120 (br.), 1686, 1479, 1386, 1014, 827, 747 cm.sup.-1;
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 2.42 (s, 1H), 4.18 (d,
1H, J=12.9 Hz), 5.56 (d, 1H, J=12.9 Hz), 7.36 (m, 3H), 7.43 (m,
2H), 7.61 (m, 1H), 7.80 (dd, 1H, J=2.1 Hz, 8.7 Hz); MS (EI) m/e
(rel intensity) 386 (M.sup.+, 45), 357 (100); Anal. Calcd. For
C.sub.17H.sub.12N.sub.4BrCl.0.5H.sub.2O: C, 51.65; H, 3.32; N,
14.18. Found C, 51.95; H, 2.97; N, 13.91.
8-Trimethylsilylacetylenyl-5-(2'-chlorophenyl)-1-methyl-4H-s-triazolo-[4,3-
-a]-1,4-benzodiazepine 22 (XLi-JY-DMH-TMS).sup.4,7,8
[0081] A mixture of 21 (7.75 g, 20 mmol), acetonitrile (600 mL),
triethylamine (500 mL) and bis(triphenylphosphine)-palladium (II)
acetate (1.2 g, 1.6 mmol) was degassed. Tri-methylsilylacetylene
(5.65 mL, 40 mmol) was then added and the solution was degassed
again. The solution was then heated to reflux for 4 hr until
analysis by TLC indicated the starting material had disappeared.
The mixture was cooled to rt and concentrated under reduced
pressure. The residue was partitioned between H.sub.2O (50 mL) and
EtOAc (2.times.200 mL). The combined organic layer was washed with
brine and dried (Na.sub.2SO.sub.4). The residue was purified by
flash chromatography on silica gel (CHCl.sub.3) to furnish the
trimethylsilyl analogue 22 (XLi-JY-DMH-TMS, 3 g, 37.0%) as white
solid: mp 265-267.degree. C.; IR (KBr) 2930, 1618, 1554, 1497,
1429, 1316, 885, 847 cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 0.24 (s, 9H), 2.65 (s, 3H), 4.15 (d, 1H, J=12.9 Hz), 5.52
(d, 1H, J=12.9 Hz), 7.35-7.45 (m, 5H), 7.61 (m, 1H), 7.72 (dd, 1H,
J=1.8 Hz, 8.4 Hz); MS (EI) m/e (rel intensity) 404 (M.sup.+, 90),
375 (100); Anal. Calcd. For C.sub.22H.sub.21N.sub.4SiCl: C, 65.33;
H, 5.24; N, 13.86. Found: C, 64.99; H, 4.98; N, 13.79.
[0082]
8-Acetyleno-5-(2'-chlorophenyl)-1-methyl-4H-s-triazolo-[4,3-a]-1,4--
benzodiazepine 23 (XLi-JY-DMH.sup.+)..sup.7 A solution of
benzodiazepine 22 (1.25 g, 31 mmol) in THF (250 mL) was cooled to
-30.degree. C. and treated with Bu.sub.4NF..times.H.sub.2O (0.97 g,
37 mmol). After the mixture was stirred for 5 min, analysis by TLC
(silica gel; EtOAc:EtOH 4:1) indicated starting material had
disappeared. Water (70 mL) was then added and the mixture was
allowed to warm to rt. The mixture was then extracted with EtOAc
(2.times.200 mL). The organic layer was washed with brine and dried
(Na.sub.2SO.sub.4). After removal of the solvent under reduced
pressure, the residue was washed successively with ethyl ether,
ethyl acetate and chloroform. After drying, the title compound 23
(XLi-JY-DMH) was obtained (1.0 g, 97.3%) as a white solid:
mp>250.degree. C. (dec); IR (KBr) 3185, 1623, 1543, 1497, 1429,
756 cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 2.65 (s,
3H), 3.17 (s, 1H), 4.18 (d, 1H, J=12.9 Hz), 5.54 (d, 1H, 12.9 Hz),
7.34 (m, 2H), 7.41-7.45 (m, 3H), 7.6 (m, 1H), 7.75 (dd, 1H, J=1.8
Hz, 8.4 Hz); MS (EI) m/e (rel intensity) 332 (M.sup.+, 78) 303
(100).
##STR00050##
[0083] Esters 37 (dm-II-30), 38 (dm-II-33) and 41 (dm-II-20) were
prepared according to the general procedure described above from
the starting acids and different alcohols, respectively. The
bromide 37 was converted into the trimethlyacetylenyl compound 39
(dm-II-35) under standard conditions (Pd-mediated, Heck-type
coupling) .sup.4,7,8 (Scheme 7).
[0084] General Procedure for Preparing the Esters.
[0085] The acid was dissolved in DMF (10 mL/mmol S.M.) and CDI (1.2
eq) was added. The reaction mixture was stirred at room temperature
for 3 h followed by addition of the alcohol (10 eq) and DBU (1 eq).
The stirring was maintained until the disappearance of all the
starting material as determined by TLC (EtOAc:EtOH 4:1). The
reaction mixture was then quenched by adding water. The solid which
precipitated was filtered and washed with ethyl ether. It was
purified by flash chromatography (EtOAc) on silica gel or neutral
aluminum oxide for ester 38.
Trifluoroethyl
8-bromo-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxylate
37 (dm-II-30)
[0086] A white solid (69.1%) from acid 27 and
2,2,2-trifluoroethanol: mp 202-204.degree. C.; IR (KBr) 3114, 1711,
1608, 1495, 1368, 1288, 1158 cm.sup.-1; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 4.10 (d, 1H, J=12.6 Hz), 4.68 (m, 1H), 4.85 (m,
1H), 6.02 (d, 1H, J=12.6 Hz), 7.41-7.54 (m, 6H), 7.62 (d, 1H, J=2.1
Hz), 7.83 (dd, 1H, J=2.1 Hz, 8.4 Hz), 7.97 (s, 1H); MS (EI) m/e
(rel intensity) 463 (M.sup.+, 14), 465 (14).
Trichloroethyl
8-bromo-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxylate
38 (dm-II-33)
[0087] A white solid (90.9%) from acid 27 and
2,2,2-trichloroethanol: mp 113-116.degree. C.; IR (KBr) 3434, 1728,
1610, 1493, 1270, 1146, 1128 cm.sup.-1; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 4.11 (d, 1H, J=12.6 Hz), 4.91 (d, 1H, J=12.0
Hz), 5.19 (d, 1H, J=12.0 Hz), 6.12 (d, 1H, J=12.6 Hz), 7.41-7.54
(m, 6H), 7.61 (d, 1H, J=2.1 Hz), 7.83 (dd, 1H, J=2.1 Hz, 8.4 Hz);
MS (EI) m/e (rel intensity) 511 (M.sup.+, 45).
Trifluoroethyl
8-trimethylsilylacetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepi-
ne-3-carboxylate 39 (dm-II-35)
[0088] A white solid (49.8%): mp 107-110.degree. C.; IR (KBr) 2961,
1734, 1611, 1560, 1497, 1251, 1159, 1120, 846 cm.sup.1; .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 0.25 (s, 9H), 4.08 (d, 111, J=12.3
Hz), 4.69 (m, 1H), 4.84 (m, 1H), 5.98 (d, 1H, J=12.3 Hz), 7.39-7.57
(m, 7H), 7.76 (dd, 1H, J=1.8 Hz, 8.4 Hz); MS (EI) m/e (rel
intensity) 481 (M.sup.+, 100).
Trifluoroethyl
8-acetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diaze-pine-3-carboxyl-
ate 41 (dm-II-20)
[0089] A white solid (36.9%) from acid 40 and
2,2,2-trifluoroethanol: mp 188-190.degree. C.; IR (KBr) 3443, 3277,
1710, 1600, 1492, 1366, 1280, 1156 cm.sup.-1; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 3.18 (s, 1H), 4.08 (d, 1H, J=12.5 Hz), 4.67 (m,
1H), 4.82 (m, 1H), 5.98 (d, 1H, J=12.5 Hz), 7.37-7.40 (m, 2H),
7.44-7.51 (m, 3H), 7.56-7.59 (m, 2H), 7.78 (dd, 1H, J=1.5 Hz, 8.5
Hz); MS (EI) m/e (rel intensity) 409 (M.sup.+, 28). Anal. Calcd.
For C.sub.22H.sub.14N.sub.3O.sub.2F.sub.3.0.25H.sub.2O: C, 63.82;
H, 3.72; N, 10.16. Found: C, 63.89; H, 3.37; N, 9.94.
##STR00051##
[0090] The bromide 1 was reacted with diethylphosphorochloridate in
the presence of sodium hydride, followed by addition of t-butyl
isocyanoacetate to provide the ester 42. This was converted into
the trimethylsilylacetyleno compound 43 under standard conditions
(Pd-mediated, Heck-type coupling)..sup.8 Treatment of 43 with
fluoride gave the title compound 44.
Procedure for XLi225
[0091] t-Butyl
8-bromo-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxylate
42. This benzodiazepine 42 was obtained in 40% yield from 1.sup.1
analogous to the literature procedure.sup.2 as a white solid. 42
(XLi223): mp: 222.degree.-223.degree. C.; IR (KBr) 2975, 2358,
1717, 1608, 1557, 1277, 1073, 908, 696, 652 cm.sup.-1; .sup.1H NMR
(CDCl.sub.3) .delta. 1.60 (s, 9H), 4.03 (d, 1H, J=12.5 Hz), 6.08
(d, 1H, J=12.4 Hz), 7.35-7.52 (m, 7H), 7.58 (d, 1H, J=2.2 Hz), 7.80
(dd, 1H, J=2.22 Hz and 8.55 Hz), 7.93 (s, 1H).
[0092]
t-Butyl-8-trimethylsilylacetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5--
a][1,4]-diazepine-3-carboxylate 43 (XLi 224)..sup.4,5,8 A mixture
of bromide 42 (1 g, 2.28 mmol, trimethylsilylacetylene (559 mg,
5.69 mmol) and bis(triphenylphosphine)-palladium-(II) acetate (55
mg, 0.073 mmol) in a mixed solvent system of CH.sub.3CN (15 mL) and
anhydrous TEA (25 mL) was heated to reflux under argon. After
stirring for 6 hours at reflux, the mixture was cooled to room
temperature and the precipitate which formed was removed by
filtration. The filtrate was concentrated under reduced pressure
and the residue was treated with a saturated aqueous solution of
NaHCO.sub.3 (20 mL), and extracted with CHCl.sub.3 (3.times.25 mL).
The combined extracts were washed with brine and dried
(Na.sub.2SO.sub.4). After removal of solvent under reduced
pressure, the residue was purified by flash chromatography (silica
gel, EtOAc) to afford 43 (XLi224) as a white solid (710 mg,
68.9%).mp: 234.degree.-236.degree. C.; IR (KBr) 2973, 2357, 2154,
1719, 1611, 1493, 1366, 1250, 1152, 1075, 946, 880 cm.sup.-1;
.sup.1H NMR (CDCl.sub.3) .delta. 0.23 (s, 9H), 1.64 (s, 9H), 4.05
(d, 1H, J=12.7 Hz), 6.06 (d, 1H, J=12.4), 7.37-7.53 (m, 7H), 7.73
(dd, 1H, J=1.95 and 8.25 Hz), 7.92 (s, 1H); MS (EI) m/e (relative
intensity) 427 (M.sup.+, 76), 412 (5), 381 (55), 353 (100) 303
(10), 287 (7).
[0093] t-Butyl
8-acetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxyla-
te 44 (XLi 225)..sup.7 A solution of 43 (128 mg, 0.281 mmol), in
THF (15 mL) was treated with Bu.sub.4NF.H.sub.2O (100.04 mg, 0.38
mmol). The mixture which resulted was allowed to stir for 5 min at
room temperature after which the mixture was added to H.sub.2O (10
mL) and extracted with EtOAc (3.times.15 mL). The combined organic
extracts were washed with brine (15 mL) and dried
(Na.sub.2SO.sub.4). After removal of solvent under reduced
pressure, the residue was purified by a wash column (silica gel,
EtOAc) to furnish 44 (XLi225) (92 mg, 85.4%) as a white solid: mp:
221.degree.-223.degree. C.; IR (KBr) 3159, 3107, 2092, 1721, 1606
cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta. 1.62 (s, 9H), 3.21 (s,
1H), 4.12 (d, 1H, J=10.2 Hz), 6.07 (d, 1H, J=12.5 Hz), 7.35-7.53
(m, 7H), 7.73 (dd, 1H, J=1.8 Hz and 8.3 Hz), 7.92 (s, 1H).
##STR00052##
[0094] 7-Bromo-2'-fluorobenzodiazepine 13 was hydrolyzed with aq 2
N sodium hydroxide in EtOH and acidified to pH 4 by adding 1 N HCl
to afford the acid 45. The acid, obtained from the ester 13, was
stirred with CDI in DMF, followed by stirring with trifluoroethanol
and DBU to provide the ester 46 (JYI-049). This material 46 was
heated with trimethysilylacetylene in a Heck-type coupling
reaction.sup.8 to provide the trimethylsilyl analog 47 (JYI-053).
The silyl group was removed from 47 on treatment with
tetrabutylammonium fluoride to furnish 48 (JYI-059) in 70%
yield.
Procedure:
[0095]
8-Bromo-6-(2'-fluorophenyl)-4H-benzo[f]imidazo[1,5-a][1,4]diaze-pin-
e-3-carboxylic acid 45. The ester 13 (1.0 g, 2.36 mmol) was
dissolved in EtOH (80 mL) and 2 N aq NaOH (8 mL) was added to the
solution. The mixture was stirred at rt for 4 hours. After the EtOH
was removed under reduced pressure, the solution was allowed to
cool. The pH value was adjusted to 4 by adding 1 N HCl dropwise.
The mixture was filtered and the solid was washed with cold water
and ethyl ether. The solid was dried to afford 45 (0.96 g, 97%) as
a white solid: mp 280.degree. C. (dec); IR (KBr) 3419, 1740, 1611,
1491 cm.sup.-1; .sup.1H NMR (DMSO-d.sub.6) .delta. 4.11 (bs, 1H),
5.99 (bs, 1H), 7.20 (t, 1H, J=8.5 Hz), 7.32 (t, 1H, J=7.5 Hz), 7.38
(d, 1H, J=1.8 Hz), 7.55 (m, 2H), 7.84 (d, 1H, J=8.7 Hz), 7.95 (dd,
1H, J=8.6, 1.9 Hz), 8.35 (s, 1H). MS (EI) m/e (relative intensity)
400 (72), 399 (85), 381 (100), 355 (82).
[0096]
Trifluoroethyl-8-bromo-6-(2'-fluorophenyl)-4H-benzo[f]imidazo[1,5-a-
][1,4]diazepine-3-carboxylate 46 (JYI-049). The carboxylic acid 45
(0.89 g, 2.23 mmol) was dissolved in dry DMF (20 mL), after which
CDI (0.72 g, 4.45 mmol) was added at rt and the mixture was stirred
for 12 hours. The trifluoroethanol (0.49 mL, 6.68 mmol) in DMF (1
mL) and DBU (0.37 mL, 2.45 mmol) in DMF (1 mL) were then added to
the mixture and stirring continued overnight. The solvent was
evaporated under reduced pressure and the residue was purified by
flash chromatography (silica gel, hexanes/EtOAc: 3/1) to afford 46
(JYI-049, 0.81 g, 76%) as a white solid: mp 223-224.degree. C.;
IR(CHCl.sub.3) 3063, 1732, 1611, 1492 cm.sup.-1; .sup.1H NMR
(CDCl.sub.3) .delta. 4.16 (bs, 1H), 4.80 (bs, 2H), 6.07 (bs, 1H),
7.06 (dt, 1H, J=8.3, 0.9 Hz), 7.30 (m, 2H), 7.48 (m, 2H), 7.68 (dt,
1H, J=7.6, 1.8 Hz), 7.80 (dd, 1H, J=8.6, 2.1 Hz), 8.11 (s, 1H). MS
(EI) m/e (relative intensity) 483 (38), 383 (64), 355 (100). Anal.
Calcd. for C.sub.20H.sub.12N.sub.3O.sub.2F.sub.4Br: C, 49.81; H,
2.51; N, 8.71. Found: C, 49.97; H, 2.44; N, 8.68.
[0097]
Trifluoroethyl-8-trimethylsilylacetylenyl-6-(2'-fluorophenyl)-4H-be-
nzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxylate 47 (JYI-053). A
mixture of bromide 46 (JYI-049, 482 mg, 1.0 mmol),
trimethylsilylacetylene (0.28 mL, 2.0 mmol) and
bis(triphenylphosphine)palladium (II) acetate (75 mg, 0.1 mmol) in
a mixed solvent system of CH.sub.3CN (25 mL) and anhydrous
triethylamine (25 mL) was heated to reflux under argon. After
stirring for 12 h at reflux, the mixture was cooled to room
temperature and the precipitate which formed was removed by
filtration. The filtrate was concentrated under reduced pressure
and the residue was treated with a saturated aq solution of
NaHCO.sub.3 (40 mL), and extracted with CHCl.sub.3 (3.times.100
mL). The combined organic extracts were washed with brine
(2.times.50 mL) and dried (Na.sub.2SO.sub.4). After removal of
solvent under reduced pressure, the residue was purified by flash
chromatography (silica gel, hexanes/EtOAc: 3/1) to afford 47
(JYI-053, 360 mg, 76%) as a gray solid: mp 220-221.degree. C.;
IR(CHCl.sub.3) 2960, 1741, 1612, 1496 cm.sup.-1; .sup.1H NMR
(CDCl.sub.3) .delta. 0.25 (s, 9H), 4.12 (bs, 1H), 4.82 (bs, 2H),
6.10 (bs, 1H), 7.06 (t, 1H, J=8.3 Hz), 7.30 (m, 1H), 7.48 (m, 2H),
7.56 (d, 1H, J=8.3 Hz), 7.67 (m, 1H), 7.73 (dd, 1H, J=8.3, 1.8 Hz),
8.02 (s, 1H); MS (EI) m/e (relative intensity) 499 (52), 399 (45),
371 (100), 235 (21), 178 (36). Anal. Calcd. for
C.sub.25H.sub.21N.sub.3O.sub.2F.sub.4Si: C, 60.11; H, 4.24; N,
8.41. Found: C, 60.27; H, 4.22; N, 8.33.
[0098]
Trifluoroethyl-8-acetyleno-6-(2'-fluorophenyl)-4H-benzo[f]imidazo-[-
1,5-a][1,4]diazepine-3-carboxylate 48 (JYI-059). A solution of 47
(JYI-053, 475 mg, 1.0 mmol) in THF (15 mL) was treated with
Bu.sub.4NF (2 mL, 1.0M solution in THF). The mixture, which
resulted, was allowed to stir for 5 min at room temperature after
which the mixture was added to H.sub.2O (5 mL) and extracted with
EtOAc (3.times.10 mL). The combined organic extracts were washed
with brine (2.times.10 mL) and dried (Na.sub.2SO.sub.4). The
solvent was removed under reduced pressure and the residue was
recrystallized from ethyl acetate/hexanes to afford 48 (JYI-059,
299 mg, 70%) as a pale yellow solid: mp 192-193.degree. C.;
IR(CHCl.sub.3) 3295, 3052, 1741, 1612, 1494, 1277, 1159 cm.sup.-1;
.sup.1H NMR (CDCl.sub.3) .delta. 3.14 (s, 1H), 4.17 (bs, 1H), 4.78
(bs 2H), 4.47 (s, 1H), 6.05 (bs, 1H), 7.05 (dt, 1H, J=8.3, 0.8 Hz),
7.30 (m, 1H), 7.48 (m, 2H), 7.60 (d, 1H, J=8.3 Hz), 7.68 (dt, 1H,
J=7.6, 1.8 Hz), 7.76 (dd, 1H, J=10.1, 1.8 Hz), 8.02 (s, 1H); MS
(EI) m/e (relative intensity) 427 (37), 327 (26), 299 (100), 178
(50). Anal. Calcd. for C.sub.22H.sub.13N.sub.3O.sub.2F.sub.4: C,
61.83; H, 3.07; N, 9.83. Found: C, 61.94; H, 3.03; N, 9.68.
##STR00053##
[0099] Ethyl amido oxime (59.5 mg, 0.676 mmol) was added to a
stirred suspension of powdered 4 .ANG. molecular sieves (75 mg) in
anhydrous THF (15 mL) under nitrogen. After the mixture was stirred
at rt for 10 min, NaH (27 mg of 60% in mineral oil, 0.676 mmol) was
added to the mixture. After the mixture was stirred for a further
30 min, a solution of the forgoing ester 7 (XHeII-053, 120 mg,
0.338 mmol) in THF (20 mL) was added. The mixture which resulted
was heated to reflux for 8 hr. It was cooled to rt, after which
acetic acid (40.6 mg, 0.676 mmol) was added. After the solution was
stirred for 10 min, the mixture was filtered through celite. The
filtrate was diluted with CH.sub.2Cl.sub.2 (50 mL) and washed with
water, brine and dried (K.sub.2CO.sub.3). Evaporation of the
solvent under reduced pressure afforded a pale yellow solid, which
was purified by flash column chromatography (silica gel,
EtOAc/hexane, 2:3) to furnish 51 as a white solid (PS-1-26, 52 mg,
40%). mp: 221-222.degree. C.; IR (KBr) 3297, 3105, 1631, 1570,
1495, 1310, 938 cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta. 8.07
(s, 1H), 7.80 (dd, 1H, J=8.4 Hz, J=1.8 Hz), 7.64-7.60 (m, 2H),
7.53-7.37 (m, 5H), 6.12 (d, 1H, J=12.9 Hz), 4.21 (d, 1H, J=12.9
Hz), 3.20 (s, 1H), 2.88 and 2.83 (ABq, 2H, J=7.6 Hz), 1.41 (t, 3H,
J=7.6 Hz); .sup.13C NMR (CDCl.sub.3) .delta. 171.8, 170.6, 168.8,
139.1, 136.6, 135.8, 135.4 (2C), 135.1, 130.7, 129.3 (2C), 128.3
(2C), 128.1, 124.7, 122.7, 121.6, 81.2, 80.0, 44.7, 19.7, 11.5; MS
(m/z) 379 (100).
[0100] This compound 49 (PS-1-27) was obtained in 47% yield from 5
(dm-1-70) analogous to the procedure employed above as a white
solid. mp: 210.degree. C.; IR (KBr) 3106, 1631, 1563, 1493, 1147,
931, 698 cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta. 8.06 (s, 1H),
7.84 (dd, 1H, J=8.6 Hz, J=2.25 Hz), 7.63-7.38 (m, 7H), 6.13 (d, 1H,
J=12.9 Hz), 4.21 (d, 1H, J=12.9 Hz), 3.20 (s, 1H), 2.88 and 2.83
(ABq, 2H, J=7.6 Hz), 1.41 (t, 3H, J=7.6 Hz); MS (m/z) 435
(100).
[0101] To the suspension of compound 49 (PS-1-27, 0.5 g, 1.15 mmol)
in acetonitrile (30 mL) and triethylamine (80 mL) was added
bis(triphenylphosphine)palladium (II) acetate (0.086 g, 0.115
mmol). The solution was degassed and trimethylsilylacetylene (0.33
mL, 2.3 mmol) added. The mixture was heated to reflux and stirred
overnight. After removal of the solvent, the residue was dissolved
in CH.sub.2Cl.sub.2 and washed with a saturated aqueous solution
NaHCO.sub.3 and brine. The organic layer was dried
(Na.sub.2SO.sub.4) filtered and concentrated under vacuum. The
residue was purified by flash column chromatography (EtOAc:hexane
2:3) to furnish the trimethylsilyl analog 50 (PS-1-28, 380 mg, 73%)
as a pale yellow solid: mp: 193-194.degree. C.; IR (KBr) 3106,
2960, 2149, 1630, 1567, 1493, 938, 851, 701 cm.sup.1; .sup.1H NMR
(300 Hz, CDCl.sub.3) .delta. 8.07 (s, 1H), 7.78 (dd, 1H, J=1.86,
8.34 Hz), 7.61-7.38 (m, 7H), 6.11 (d, J=12.78 Hz), 4.19 (d, J=12.78
Hz), 2.88 and 2.83 (ABq, 2H, J=7.56 Hz), 1.41 (t, 3H, J=7.56 Hz),
0.25 (s, 9H).
##STR00054##
[0102] The bromide 20 available from references 9 and 10 was
reacted with trimethylsilyacetylene in the presence of a palladium
catalyst to provide trimethylsilyl analog 52. This product was
methylated with methyl iodide/sodium hydride to give the N-methyl
benzodiazepine 54 (XLi 351). This was subjected to
fluoride-mediated desilation to furnish 53 (XLi 350) and 55 (XLi
352).
Procedure for XLi 350 and XLi 352
[0103]
7-Trimethylsilylacetyleno-5-phenyl-(2'-chlorophenyl)1,3-dihydrobenz-
o[e]-1,4-diazepin-2-one 52 (XLi 343)..sup.4,5,8 A mixture of
20.sup.1 (500 mg, 1.43 mmole) available from references 9 and 10 in
triethyl amine (10 mL) and CH.sub.3CN (16 mL) with
trimethyl-silylacetylene (126 mg, 1.28 mmole) and
bis(triphenylphosphine)palladium (II) acetate (64.3 mg, 0.086 mmol)
was heated to reflux under nitrogen. After 6 hours, the reaction
mixture was cooled to room temperature and filtered. The filtrate
was concentrated under vacuum and the residue was treated with a
saturated aqueous NaHCO.sub.3 solution (15 mL), and extracted with
CH.sub.2Cl.sub.2 (3.times.20 mL). The organic layers were combined
and washed with brine and dried (Na.sub.2SO.sub.4). After removal
of solvent under reduced pressure, the residue was purified via
flash chromatography (silica gel, EtOAc/hexanes:1/1) to furnish 52
as a yellow powder (310 mg, 59%): mp: 225.8-228.2.degree. C.; IR
(KBr) 2953, 2358, 1685, 1616, 1490, 1328, 1248, 1058, 1011, 841,
746 cm.sup.-1, .sup.1H NMR (CDCl.sub.3) .delta. 0.21 (s, 9H), 4.38
(s, 2H), 7.41 (d. 1H, J=8.37 Hz), 7.19-7.52 (br, 7H), 8.11 (s, 1H);
MS (EI) m/e (relative intensity) 366 (M.sup.+, 100), 331(59),
229(18), 161 (26).
[0104]
7-Acetyleno-5-phenyl-(2'-chlorophenyl)-1,3-dihydro-benzo[e]-1,4-dia-
zepin-2-one 53 (XLi 350):.sup.7 A solution of 52 (150 mg, 0.408
mol) in THF (30 mL) was treated with tetrabutylammonium fluoride
(1M in THF). The mixture was stirred for 20 minutes at room
temperature before water (30 mL) was added. The mixture was then
extracted with EtOAc (3.times.30 mL). The combined organic extracts
were washed with brine and dried over (Na.sub.2SO.sub.4). The
solvent was removed under vacuum and the residue which resulted was
passed through a wash column (silica gel, EtOAc/hexanes:4/1) to
give 55 as light yellow crystals (110 mg, 95.2%); mp: 215.degree.
C.; IR (KBr) 3290, 1685, 1615, 1491, 1328, 731 cm.sup.-1, .sup.1H
NMR (CDCl.sub.3) .delta. 3.06 (s, 1H), 4.40 (s, 3H), 7.03-7.61 (m,
7H), 7.58-7.86 (m, 2H), 7.99 (s, 1H); MS (EI) m/e (relative
intensity) 294 (M.sup.+, 100), 266(75), 265(87), 259(83), 231(40),
201(24), 176 (23).
[0105]
1-Methyl-7-trimethylsilylacetyleno-5-phenyl-(2'-chlorophenyl)-1,3-d-
ihydro-benzo[e]-1,4-diazepin-2-one 54 (XLi 351)..sup.7 A mixture of
52 (300 mg, 0.82 mmol) was dissolved in dry THF (40 mL) at
0.degree. C. and NaH (60% in mineral oil, 50 mg, 1.25 mmol) was
added to the solution in one portion. The slurry was then stirred
for 20 min at 0.degree. C. and CH.sub.3I (139 mg, 0.98 mmol) was
added to the mixture and it was warmed up to room temperature.
After the mixture stirred for 3 hours at room temperature, the THF
was then removed under reduced pressure. The residue was purified
by flash chromatography [hexanes/EtOAc (1:4)] to provide the title
compound 54 (260 mg, 83%) as a white solid: mp: 196.9-198.degree.
C.; IR (KBr) 2953, 1676, 1611, 1489, 1346, 1125, 1078, 913, 742
cm.sup.-1; .sup.1HNMR (CDCl.sub.3) 8 (ppm) 0.21 (s, 9H) 3.46 (s,
3H), 3.54 (d, 1H, J=10.9 Hz), 4.60 (d, 1H. J=10.8 Hz), 7.20-7.43
(m, 5H), 7.58-7.65 (m, 3H). MS (EI) m/e (relative intensity) 380
(M.sup.+, 8), 366(10), 308(100), 280(88), 273(97), 245 (61).
[0106]
1-Methyl-7-acetyleno-5-phenyl-(2'-chlorophenyl)-1,3-dihydro-benzo[e-
]-1,4-diazepin-2-one 55 (XLi 352):.sup.7 A solution of 54 (100 mg,
0.262) in THF (30 mL) was treated with tetrabutylammonium fluoride
(1M in THF). The mixture was stirred for 20 minutes at room
temperature before water (30 mL) was added. The mixture was then
extracted with EtOAc (3.times.30 mL). The combined organic extracts
were washed with brine and dried (Na.sub.2SO.sub.4). The solvent
was removed under vacuum and the residue which resulted was passed
through a wash column (silica gel, EtOAc/hexanes:4/1) to give 55 as
light yellow crystals (71 mg, 90%): mp: 95.6-98.1.degree. C.; IR
(KBr) 2953, 1677, 1489, 1346, 1091, 791, 749 cm.sup.-1, .sup.1H NMR
(CDCl.sub.3) 8 (ppm) 3.05 (s, 1H), 3.46 (s, 3H), 3.83 (d, 1H,
J=10.5 Hz), 4.87 (d, 1H, J=9.33 Hz), 5.28 (s, 1H), 7.20-7.43 (m,
5H), 7.58-7.86 (m, 2H); MS (EI) m/e (relative intensity) 308
(M.sup.+, 100), 294(19), 280(82), 273(99), 249(28), 245(61),
229(29), 201(32), 189 (43).
##STR00055##
[0107]
7-Trimethylsilylacetyleno-5-(2'-fluorophenyl)-1,3-dihydrobenzo[e]-1-
,4-diazepine-2-one 56 (JYI-55). A mixture of bromide 12 (1.6 g, 5.0
mmol), trimethylsilyl-acetylene (3.0 mL, 21.0 mmol) and
bis(triphenylphosphine)palladium (II) acetate (375 mg, 0.5 mmol) in
a mixed solvent system of CH.sub.3CN (60 mL) and anhydrous
triethylamine (40 mL) was heated to reflux under argon. After
stirring for 3 h at reflux, the mixture was cooled to room
temperature and the precipitate which formed was removed by
filtration. The filtrate was concentrated under reduced pressure
and the residue was treated with a saturated aq solution of
NaHCO.sub.3 (100 mL), and extracted with CHCl.sub.3 (3.times.200
mL). The combined organic extracts were washed with brine
(2.times.100 mL) and dried (Na.sub.2SO.sub.4). After removal of
solvent under reduced pressure, the residue was purified by flash
chromatography (silica gel, hexanes/EtOAc: 2/1) to afford 56
(JYI-55, 794 mg, 47%) as a gray solid: mp 168.5-169.5.degree. C.;
IR (CHCl.sub.3) 3202, 3113, 2955, 1686, 1612, 1490 cm.sup.1;
.sup.1H NMR (CDCl.sub.3) .delta. 0.22 (s, 9H), 4.38 (s, 2H),
7.04-7.33 (m, 3H), 7.34 (s, 1H), 7.45-7.53 (m, 1H), 7.56-7.62 (m,
2H), 8.73 (bs, 1H). MS (EI) m/e (relative intensity) 350 (94), 322
(100), 167 (41), 153 (37). Anal. Calcd. for
C.sub.20H.sub.19N.sub.2OFSi: C, 68.54; H, 5.46; N, 7.99. Found: C,
68.23; H, 5.40; N, 8.34.
[0108]
7-Acetyleno-5-(2'-fluorophenyl)-1,3-dihydrobenzo[e]1,4-diazepine-2--
one 57 (JYI-60). A solution of 56 (JYI-55, 700 mg, 2.0 mmol) in THF
(200 mL) was treated with Bu.sub.4NF (2 mL, 1.0M solution in THF).
The mixture, which resulted, was allowed to stir for 5 min at room
temperature after which the mixture was added to H.sub.2O (5 mL)
and extracted with EtOAc (3.times.10 mL). The combined organic
extracts were washed with brine (2.times.10 mL) and dried
(Na.sub.2SO.sub.4). After the solvent was removed under reduced
pressure, the residue was purified by flash chromatography (silica
gel, hexanes/EtOAc: 2/1) to afford 57 (JYI-60, 400 mg, 72%) as a
pale yellow solid: mp 208-209.5.degree. C.; IR(CHCl.sub.3) 3290,
3110, 2930, 1685, 1612, 1489 cm.sup.1; .sup.1H NMR (CDCl.sub.3)
.delta. 3.04 (s, 1H), 4.40 (s, 2H), 7.06-7.28 (m, 3H), 7.38 (s,
1H), 7.44-7.51 (m, 1H), 7.59-7.62 (m, 2H), 9.43 (bs, 1H). MS (EI)
m/e (relative intensity) 278 (80), 250 (100). Anal. Calcd. for
C.sub.17H.sub.11N.sub.2OF: C, 73.37; H, 3.98; N, 10.07. Found: C,
73.64; H, 3.92; N, 9.78.
##STR00056##
[0109] 2-Amino-5-iodo-benzophenone was prepared from
p-iodonitrobenzene and phenylacetonitrile according to the
literature..sup.112-Amino-5-chloro-benzophenone was commercially
available from Acros. The benzodiazepine 60 was reacted with
diethylphosphorochloridate in the presence of sodium hydride,
followed by the addition of ethyl isocyanoacetate to provide the
ester 62 (Hz120), as shown in Scheme 13.
[0110] Ethyl
8-iodo-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxylate
62. A solution of benzodiazepine 60 (3 g, 8.3 mmol) in dry THF (36
mL) was cooled to 0.degree. C. and a 60% dispersion of sodium
hydride (0.70 g, 17.4 mmol) was added in one portion. The mixture
was allowed to warm to rt with stirring and the stirring was
continued at rt until no more bubbles were evolved. The suspension
was cooled to 0.degree. C. after which diethylphosphorochloridate
(2.29 g, 13.3 mmol) was added and this mixture was stirred for 30
min and allowed to warm to rt. The mixture was stirred for an
additional 1.5 hr. In another flask, a 60% dispersion of sodium
hydride (0.70 g, 17.4 mmol) in mineral oil was added in dry THF (36
mL) and cooled to 0.degree. C. Ethyl isocyanoacetate (1.13 g, 9.94
mmol) was added and the stirring was continued until no more
bubbles were evolved. This mixture was transferred to the above
mixture at 0.degree. C. The mixture was then stirred at rt for 6 h
and quenched with HOAc (3.2 mL). The mixture was partitioned
between EtOAc (200 mL) and H.sub.2O (50 mL). The organic layer was
washed with brine and dried (Na.sub.2SO.sub.4). After the solvent
was removed under reduced pressure, the residue was purified by
flash chromatography (silica gel, gradient elution, EtOAc:hexane
1:4, 1:1, 4:1) to provide the ester 62 (Hz120) in 43% yield as a
light brown solid. mp: 221-222.degree. C.; IR (KBr) 2977, 1717,
1608, 1489 cm.sup.-1; .sup.1H NMR (DMSO-d.sub.6) .delta. 1.31 (t,
3H, J=7.1 Hz), 4.10 (d, 1H, J=12.5 Hz), 4.29 (q, 2H, J=6.7 Hz),
5.75 (d, 1H, J=12.4 Hz), 7.40-7.50 (m, 5H), 7.63 (d, 1H, J=1.8 Hz),
7.69 (d, 1H, J=8.5 Hz), 8.13 (dd, 1H, J=1.9, 8.5 Hz), 8.36 (s, 1H);
MS (EI) m/e (relative intensity) 458 (23), 457 (M.sup.+, 100), 411
(62), 384 (29), 383 (100), 257 (29). Anal. Calcd. for
C.sub.20H.sub.16IN.sub.3O.sub.2: C, 52.53; H, 3.53; N, 9.19. Found:
C, 52.57, H, 3.73; N, 8.64.
[0111] Ethyl
8-chloro-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxylate
63. This ester 63 was obtained in 52% yield from 61 analogous to
the procedure employed above as a white solid. mp: 174-175.degree.
C. (lit..sup.12 174-175.degree. C.); .sup.1H NMR (DMSO-d.sub.6)
.delta. 1.32 (t, 3H, J=7.1 Hz), 4.13 (d, 1H, J=12.3 Hz), 4.32 (q,
2H, J=6.7 Hz), 5.76 (d, 1H, J=12.3 Hz), 7.37-7.50 (m, 6H),
7.86-8.38 (m, 2H), 8.74 (s, 1H).
##STR00057##
[0112] 6-Bromo-2-phenyl-4H-benzo[2,3-d]-1,3-oxazin-4-one 64. The
2-amino-5-bromobenzoic acid (5 g, 23.1 mmol) was treated with
benzoyl chloride (237 mL, 2.04 mol) at 140.degree. C. for 3 h.
After the reaction mixture was cooled to rt, the crystals that
formed were collected by filtration and were washed with hexanes to
provide 64 as light brown needles (6.8 g, 97%): .sup.1H NMR
(CDCl.sub.3) .delta. 7.51-7.2 (m, 4H), 7.9 (dd, 1H, J=2.3, 8.6 Hz),
8.30-8.33 (m, 2H), 8.8 (d, 1H, J=2.2 Hz); .sup.13C NMR (CDCl.sub.3)
.delta. 158.19, 157.35, 145.75, 139.58, 132.82, 130.97, 129.77,
128.82, 128.73, 128.29, 121.37, 118.27; MS (EI) m/e (relative
intensity) 303 (M.sup.+, 36), 301 (M.sup.+, 36), 259 (14), 257
(14), 226 (6), 224 (6), 178 (9), 170 (9), 168 (9), 151 (4), 105
(100).
[0113] 4-Bromo-2-(2'-thienylcarbonyl)-N-benzoylaniline 66 and
bis-(2'-thienyl)-[5-bromo-2-(N-benzoyl)-amino]phenylmethanol 65.
The benzo-xazinone 64 (5.0 g, 16.6 mmol) was dissolved in dry THF
(250 mL) and cooled to -78.degree. C. for 45 min. The
2-thienyllithium (18.21 mL of 1M solution in THF) was added
dropwise over 35 min and the reaction was stirred at -78.degree. C.
for 1.2 h. Saturated aq NH.sub.4Cl solution (25 mL) and Et.sub.2O
(30 mL) were then added. The organic layer was separated, washed
with brine and dried (MgSO.sub.4). The solvent was removed under
reduced pressure, and the residue was purified via flash
chromatography (silica gel, hexanes/EtOAc: 1:0, 49:1, 20:1, 11:1,
5:1) to provide 66 as yellow crystals and the alcohol 65. 66:
.sup.1H NMR (CDCl.sub.3) .delta. 7.23 (dd, 1H), 7.52-7.56 (m, 3H),
7.66 (dd, 1H, J=0.99, 3.8 Hz), 7.82 (d, 1H, J=5.0 Hz), 7.99-8.02
(m, 3H), 7.75 (d, 1H, J=9.0 Hz), 11.2 (s, 1H); .sup.13C NMR
(CDCl.sub.3) .delta. 188.82, 165.45, 143.24, 138.79, 136.57,
135.90, 135.51, 134.25, 134.03, 132.17, 128.81, 128.31, 127.26,
125.65, 123.45, 114.95; MS (EI) m/e (relative intensity) 387
(M.sup.+, 12), 385 (M.sup.+, 12), 276 (18), 274 (18), 201 (7), 172
(7), 105 (100). 65: .sup.1H NMR (CDCl.sub.3) .delta. 4.20 (s, 1H),
6.82 (s, 2H), 6.96-7.01 (m, 3H), 7.33-7.38 (m, 7H), 7.65 (d, 2H,
J=7.23 Hz), 8.43 (d, 1H, J=8.8 Hz), 9.92 (s, 1H); .sup.13C NMR
(CDCl.sub.3) .delta. 165.04, 148.94, 136.44, 135.49, 134.49,
132.34, 131.59, 131.40, 128.40, 127.20, 126.89, 126.58, 124.18,
116.00, 79.35, 76.92, 76.50; MS (EI) m/e (relative intensity) 471
(M.sup.+, 54), 469 (M.sup.+, 51), 453 (100), 451 (93), 348 (98),
346 (92), 316 (54), 314 (58), 282 (20), 280 (19), 267 (88), 235
(12), 234 (12), 223 (15), 222 (17), 201 (56), 173 (20), 172 (12),
158 (10), 129 (10).
[0114] 5-Bromo-2-(2'-thienylcarbonyl)aniline 67. The amide 66 (2 g,
635 mmol) was dissolved in EtOH (150 mL) and 20% NaOH solution (30
mL) was added. The mixture was heated to reflux for 5 h and the
EtOH was removed under reduced pressure. The mixture was extracted
with EtOAc and the organic phases were combined, washed with brine
and dried (Na.sub.2SO.sub.4). The solvent was removed under reduced
pressure, and the residue was purified via a wash column (silica
gel, hexanes/EtOAc: 11:1 to 4:1) to provide 67 as a bright yellow
solid: .sup.1H NMR (DMSO-d.sub.6) .delta. 6.28 (br s, 2H), 6.82 (s,
1H), 6.90 (s, 1H), 7.26 (dd, 1H, J=3.8, 5.0 Hz), 7.42 (dd, 1H,
J=2.4, 8.9 Hz), 7.61 (dd, 1H, J=1.1, 3.8 Hz), 7.69 (dd, 1H, J=2.4
Hz), 8.04 (dd, 1H, J=1.1, 5.0 Hz); .sup.13C NMR (DMSO) .delta.
187.42, 150.09, 143.87, 136.46, 134.75, 134.41, 133.93, 128.78,
119.36, 119.17, 104.95; MS (EI) m/e (relative intensity) 283
(M.sup.+, 59), 282 (M.sup.+, 87), 281 (M.sup.+, 59), 280 (M.sup.+,
79), 250 (23), 248 (23), 201 (13), 199 (49), 197 (48), 172 (25),
170 (23), 145 (13), 140 (1), 111 (100), 101 (33).
[0115] 4-Bromo-2-(2'-thienylcarbonyl)-N-bromoacetylaniline 68. The
thienylaniline 67 (3.3 g, 11.7 mmol) and NaHCO.sub.3 (2.9 g, 34.5
mmol) were suspended in dry CHCl.sub.3 (180 mL) and cooled to
0.degree. C. A solution of bromoacetyl bromide (1.12 mL, 12.9 mmol)
in dry CHCl.sub.3 (30 mL) was added dropwise over 20 min at
0.degree. C. and the mixture was stirred at rt for 3 h. The
CHCl.sub.3 solution was then washed with aq NaHCO.sub.3 (5%) and
dried (Na.sub.2SO.sub.4). The CHCl.sub.3 was removed under reduced
pressure, and Et.sub.2O was added to the flask. The solution was
sonicated and filtered to provide 68 as a light solid: mp:
144.0-146.5.degree. C.; .sup.1H NMR (CDCl.sub.3) .delta. 4.01 (s,
2H), 7.23-7.26 (m, 1H), 7.24 (d, 1H), 7.65 (d, 1H), 7.74 (d, 1H),
7.84 (d, 1H), 8.46 (d, 1H), 10.85 (br s, 1H); MS (EI) m/e (relative
intensity) 405 (M.sup.+, 69), 404 (40), 403 (M.sup.+, 100), 401
(M.sup.+, 66), 324 (39), 322 (38), 310 (33), 308 (33), 292 (32),
283 (65), 282 (72), 281 (65), 280 (67), 266 (10), 264 (10), 250
(34), 248 (35), 226 (55), 224 (55), 201 (43), 199 (27), 197 (27),
173 (32), 111 (73).
[0116] 7-Bromo-5-(2'-thienyl)-1,3-dihydrobenzo[e][1,4]diazepine 69
(JC184). The bromoacetyl amide 68 (0.236 g, 0.586 mmol) was
dissolved in a saturated solution of anhydrous ammonia in MeOH (50
mL) and the mixture was heated to reflux for 6 h. After the MeOH
was removed under reduced pressure, EtOAc was added to the residue.
The solution was sonicated and then filtered to provide 69 (JC184)
as a light solid: MS (EI) m/e (relative intensity) 322 (M.sup.+,
54), 320 (M.sup.+, 53), 294 (100), 292 (98), 211 (24), 185 (31),
140 (21). The material was used directly in the next step.
[0117] 7-Trimethylsilylacetylenyl-5-(2'
thienyl)-1,3-dihydrobenzo[e][1,4]diazepine 70 (JC207). A mixture of
69 (1 g, 3.12 mmol) in CH.sub.3CN (20 mL) and Et.sub.3N (30 mL) was
degassed and heated to reflux under nitrogen.
Bis(triphenylphosphine)-palladium (II) acetate (0.26 g, 0.347 mmol)
was then quickly added, followed by the addition of TMS acetylene
(0.76 g, 7.78 mmol). The mixture was stirred at reflux for 4 h and
the solvent was removed under reduced pressure. Water (25 mL) and
EtOAc (25 mL) were added to the residue and the mixture was
filtered through celite to remove the organometallic species. The
filtrate was then extracted with EtOAc and the organic phases were
combined, washed with brine and dried (Na.sub.2SO.sub.4). The
solvent was removed under reduced pressure and the residue was
purified via flash chromatography (silica gel, hexanes/EtOAc: 11:1,
5:1) to provide 70 (JC207) as a light yellow solid: mp:
198.5-201.degree. C.; MS (EI) m/e (relative intensity) 338
(M.sup.+, 68), 337 (M.sup.+, 28), 310 (100), 295 (13), 161 (13),
147 (33), 105 (17). The material was used directly in the next
step.
[0118]
7-Acetylenyl-5-(2''-thienyl)-1,3-dihydrobenzo[e][1,4]diazepine 72
(JC208). A solution of 70 (150 mg, 0.457 mmol) in THF (30 mL) was
treated with tetrabutylammonium fluoride (1M in THF) at 0.degree.
C. for 5 minutes. Water (20 mL) was subsequently added to quench
the reaction and the THF was removed under reduced pressure. The
remaining aq solution was then extracted with EtOAc and the organic
phases were combined, washed with brine and dried
(Na.sub.2SO.sub.4). Upon removal of the solvent, Et.sub.2O was
added to the residue which was sonicated and then filtered to
provide the title compound 72 (JC208, 111 mg, 91%) as an ivory
colored solid: mp: 214-216.degree. C.; MS (EI) m/e (relative
intensity) 266 (M.sup.+, 61), 265 (M.sup.+, 30), 238 (100), 237
(49), 210 (13), 209 (10), 164 (6), 153 (7), 139 (7). This material
was used in the next step.
[0119]
1-N-methyl-7-trimethylsilylacetylenyl-5-(2''-thienyl)-1,3-dihydrobe-
nzo[e][1,4]diazepine 71 (JC209). Thiophere 70 (500 g, 1.52 mmol)
was dissolved in dry THF (25 mL) at 0.degree. C. and NaH (60% in
mineral oil, 76 mg, 1.50 mmol) was added to the solution in one
portion. After the mixture was stirred at 0.degree. C. for 30 min,
MeI (0.14 mL, 2.25 mmol) was added and the ice bath was allowed to
warm to rt. The mixture was allowed to stir for 3 h and the THF was
then removed under reduced pressure. The residue was purified via
flash chromatography (silica gel, hexanes/EtOAc 8:1, 4:1) to
provide the title compound 71 (JC209) as a white solid: mp:
171.3-173.6.degree. C.; .sup.1H NMR (CDCl.sub.3) .delta. 0.26 (br
s, 9H), 3.38 (s, 3H), 4.71 (d, 1H), 7.09 (dd, 1H, J=3.7, 5.0 Hz),
7.17 (dd, 1H, J=1.1, 3.7 Hz), 7.30 (s, 1H), 7.49 (dd, 1H, J=1.1,
5.0 Hz), 7.65 (dd, 1H, J=2.0, 8.5 Hz), 7.75 (d, 1H); .sup.13C NMR
(CDCl.sub.3) 8 (CDCl.sub.3) .delta. 170.12, 163.22, 143.65, 143.14,
134.69, 133.12, 131.38, 130.14, 127.77, 127.47, 121.01, 119.10,
103.01, 95.66, 56.38, 34.67; MS (EI) m/e (relative intensity) 352
(M.sup.+, 71), 351 (M.sup.+, 60), 337 (10), 324 (100), 309 (24),
168 (28), 154 (38).
[0120]
1-N-methyl-7-acetyleno-5-(2'-thienyl)-1,3-dihydrobenzo[e][1,4]diaze-
pine 73 (JC222). The same procedure for preparing 72 (JC208) was
applied to 73 (JC222) and a very light brown solid resulted: mp:
218.3-220.4.degree. C.; .sup.1H NMR (CDCl.sub.3) .delta. 3.16 (s,
1H), 3.39 (s, 3H), 3.78 (d, 1H, J=11.07 Hz), 4.72 (d, 1H, J=5.9
Hz), 7.08 (dd, 1H, J=3.8, 5.0 Hz), 7.31 (d, 1H, J=8.6 Hz), 7.49
(dd, 1H, J=1.0, 5.0 Hz), 7.67 (dd, 1H, J=2.0, 8.5 Hz), 7.79 (d, 1H,
J=1.9 Hz); .sup.13C NMR (CDCl.sub.3) .delta.171.04, 170.07, 163.12,
143.49, 134.79, 133.50, 131.34, 130.25, 127.85, 127.46, 121.16,
117.99, 81.83, 78.30, 56.34, 34.69. MS (EI) m/e (relative
intensity) 281 (13), 280 (M.sup.+, 60), 279 (51), 253(19), 252
(100), 251(2), 235 (11), 209 (10).
[0121] Ethyl
8-bromo-6-(2'-thienyl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxyl-
ate 74 (JC217). Dry THF (30 mL) was added to a flask containing the
benzodiazepine 69 (1.27 g, 3.96 mmol) and the solution was allowed
to cool to 0.degree. C. and NaH (60% in mineral oil, 0.191 g, 4.76
mmol) was quickly added. The mixture was stirred for 30 min at
0.degree. C. and then removed from an ice bath to stir another 1 h
at rt. Prior to adding CIPO(OEt).sub.2 (1.06 g, 6.35 mmol), the
mixture was again pre-cooled to 0.degree. C. The solution was
stirred another 3 h as the ice bath warmed to rt. Meanwhile, dry
THF (10 mL) was added to a second flask containing NaH (60% in
mineral oil, 0.229 g, 5.72 mmol). After the second mixture was
cooled to 0.degree. C., CNCH.sub.2CO.sub.2Et was added dropwise and
the solution continued to stir for 30 min at 0.degree. C. After
both reaction mixtures were again pre-cooled to 0.degree. C., the
two solutions were combined under Ar via cannula and the solution
stirred at rt overnight. The reaction was quenched with ice water
and worked up with EtOAc, and the combined organic phases were
washed with brine and dried (Na.sub.2SO.sub.4). The solvent was
removed under reduced pressure and the residue was purified via
flash chromatography (silica gel, hexanes:EtOAc 4:1, 1:1, 1:3) to
provide the title compound 74 (JC217) as an ivory solid (500 mg,
30% yield): mp: 204.0-205.3.degree. C.; .sup.1H NMR (CDCl.sub.3)
.delta. 1.45 (t, 3H, J=7.1, 14.3 Hz), 4.07 (d, 1H, J=8.8 Hz), 4.44
(dd, 2H, J=3.8, 4.7 Hz), 5.98 (d, 1H, J=12.8 Hz), 7.05 (d, 1H,
J=1.0 Hz), 7.07 (s, 1H), 7.46-7.49 (m, 2H), 7.83 (dd, 1H, J=2.2,
8.5 Hz), 7.91 (s, 1H), 7.96 (d, 1H, J=2.2 Hz): MS (EI) m/e
(relative intensity) 418 (M.sup.+, 15), 417 (M.sup.+, 68), 416
(M.sup.+, 15), 415 (M.sup.+, 64), 407 (22), 344 (26), 343 (100),
342 (30), 341 (93), 293 (15), 291 (21), 262 (18), 235 (15), 211
(12), 154 (10), 127 (11).
[0122] Ethyl
8-trimethylsilylacetylenyl-6-(2-thienyl)-4H-benzo[f]imidazo[1,5-a][1,4]di-
azepine-3-carboxylate 75 (JC220). The same procedure for preparing
70 (JC207) was applied to 75 (JC220) and an ivory colored solid
resulted: .sup.1H NMR (CDCl.sub.3) .delta. 0.29 (s, 9H), 1.45 (t,
3H, J=7.1, 14.3 Hz), 4.0 (d, 1H, J=18.1 Hz), 4.45 (dd, 2H, J=7.2,
8.5 Hz), 5.97 (d, 1H, J=12.8 Hz), 7.06-7.11 (m, 2H), 7.49 (dd, 1H,
J=1.2, 5.0 Hz), 7.52 (d, 1H, J=8.3 Hz), 7.77 (dd, 1H, J=1.9, 8.3
Hz), 7.90 (d, 1H, J=1.8 Hz), 7.93 (s, 1H). MS (EI) m/e (relative
intensity) 433 (M.sup.+, 74), 387 (49), 359 (100), 277 (28), 262
(19), 235 (24), 172 (19), 129 (17).
[0123] Ethyl
8-acetyleno-6-(2'-thienyl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carb-
oxylate 76 (JC221). The same procedure for preparing 72 (JC208) was
applied to 76 (JC221) and an ivory colored solid resulted: mp:
>198.degree. C.; .sup.1H NMR (CDCl.sub.3) .delta. 1.43 (t, 3H,
J=4.3, 11.4 Hz), 3.25 (s, 1H), 4.10 (d, 1H, J=12.8 Hz), 4.40-4.49
(m, 2H), 5.99 (d, 1H, J=12.9 Hz), 7.50 (d, 1H, J=5.0 Hz), 7.56 (d,
1H, J=8.3 Hz), 7.81 (dd, 1H, J=1.8, 8.3 Hz), 7.95 (s, 1H); MS (EI)
m/e (relative intensity) 361 (M.sup.+, 24), 315 (35), 287 (100),
237 (26), 178 (30), 153 (21), 126 (18). MS (EI) m/e (relative
intensity) 361 (M.sup.+, 29), 315 (41), 287 (100), 237 (31), 178
(40), 153 (26), 126 (21).
##STR00058##
##STR00059##
[0124] Ethyl
8-acetyleno-6-(2'-thienyl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carb-
oxylate 76 (JC221). The benzodiazepine 1 was oxidized with
3-chloroperoxybenzoic acid (mCPBA) to form 77, followed by the
addition of methylamine to afford amidine 78. N-Oxide 78 was
reacted with trimethylsilyacetylene in the presence of a palladium
catalyst to provide the trimethylsilyl analog 79 (Hz146) which was
subjected to fluoride-mediated desilation to afford 80 (Hz147), as
shown in Scheme 15. In a related route, bromide 81 was converted
into the trimethylsilylacetylene 82 (Hz141). This analog was then
transformed into target 79 (Hz146) with mCPBA or the key target
(Hz148) or treatment with fluoride (Scheme 16).
[0125]
7-Bromo-4-oxy-5-phenyl-1,3-dihydro-benzo[e][1,4]diazepin-2-one 77.
Bromide 1 (1.88 g, 5.95 mmol) was dissolved in CH.sub.2Cl.sub.2 (50
mL) and mCPBA (77% max) (1.76 g) was added at rt. The reaction
mixture was stirred overnight. The mixture was diluted with
CH.sub.2Cl.sub.2 (80 mL) and washed with a sat. solution of
NaHCO.sub.3 (50 mL), water (50 mL) and brine (40 mL). The organic
layer was dried (Na.sub.2SO.sub.4) and concentrated. The residue
was purified by flash chromatography (silica gel, EtOAc) to afford
compound 77 in 90% yield as a white solid. mp: 230-231.degree. C.
(lit..sup.13 230-231.degree. C.); .sup.1H NMR (CDCl.sub.3) .delta.
4.69 (s, 2H), 7.16 (d, 1H, J=8.7 Hz), 7.24 (d, 1H, J=2.1 Hz), 7.45
(m, 3H), 7.54 (dd, 1H, J=8.6, 2.2 Hz), 7.64 (dd, 2H, J=7.3, 3.6
Hz), 10.02 (s, 1H).
[0126]
(7-Bromo-4-oxy-5-phenyl-3H-benzo[e][1,4]diazepin-2-yl)-methyl-amine
78. Methylamine (50 mL, 2 M in THF) was added to 77 (1.9 g, 5.7
mmol) in a 100 mL round-bottom flask. The mixture was cooled to
0.degree. C. after which TiCl.sub.4 (0.54 g, 2.86 mmol) was added
dropwise. The reaction mixture was allowed to warm to rt and
stirred for 4 h. The mixture was quenched with water (5 mL),
diluted with EtOAc (100 mL) and washed with dilute NH.sub.4OH. The
organic layer was washed with water, brine and dried
(Na.sub.2SO.sub.4). After the solvent was removed under reduced
pressure, the residue was purified by flash chromatography (silica
gel, gradient elution, EtOAc, EtOAc:MeOH 10:1) to provide 78 in 86%
yield as a white solid. mp: 236-237.degree. C. (lit..sup.14
242-243.degree. C.); .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 0.21
(s, 9H), 2.91 (s, 3H), 4.17 (s, 1H), 4.85 (s, 1H), 7.13-7.66 (m,
9H).
[0127]
(7-Trimethylsilylacetylenyl-4-oxy-5-phenyl-3H-benzo[e][1,4]diazepin-
-2-yl)-methyl-amine 79 (Hz146). Trimethylsilylacetylenyl analog 79
(Hz146) was obtained in 58% yield from 78 analogous to the
procedure employed above as a light gray solid. mp: 239-240.degree.
C.; IR (KBr) 3229, 3060, 2952, 2149, 1616, 1593, 1462, 1238, 868
cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 2.89 (d, 3H,
J=4.4 Hz), 4.14 (d, 1H, J=10.6 Hz), 4.78 (d, 1H, J=10.4 Hz), 7.15
(d, 1H, J=1.7 Hz), 7.24-7.28 (m, 2H), 7.45 (m, 4H), 7.66 (m, 2H);
MS (EI) m/e (relative intensity) 361 (M.sup.+, 48), 344 (100), 303
(31), 165 (33).
[0128]
(7-Acetylenyl-4-oxy-5-phenyl-3H-benzo[e][1,4]diazepin-2-yl)-methyl--
amine 80 (Hz147). The 7-acetyleno target 80 was obtained in 90%
yield from 79 analogous to the procedure employed above as a light
yellow solid. mp: 213-214.degree. C.; IR (KBr) 3242, 3068, 2977,
1619, 1589, 1460, 1414 cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 2.89 (d, 2H, J=3.7 Hz), 2.98 (s, 1H), 4.13 (bs, 1H), 4.78
(bs, 1H), 7.18-7.71 (m, 9H); MS (EI) m/e (relative intensity) 289
(M.sup.+, 47), 272 (100), 231 (42).
[0129]
(7-Bromo-5-phenyl-3H-benzo[e][1,4]diazepin-2-yl)-methyl-amine 81
(Hz135). Bromide 81 was obtained in 70% yield from 1 analogous to
the procedure employed above as a white solid. mp: 234-235.degree.
C.; IR (KBr) 3253, 3076, 1609, 1571, 1415, 1326, 1230 cm.sup.-1;
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 2.62 (s, 3H), 3.56 (bs,
1H), 4.68 (bs, 1H), 6.34 (s, 1H), 7.17 (d, 1H, J=8.7 Hz), 7.36-7.81
(m, 7H); MS (EI) m/e (relative intensity) 329 (80), 328 (M.sup.+,
100), 327 (82), 326 (92), 220 (38), 219(48), 218(46), 205 (38).
[0130]
(7-Trimethylsilylacetylenyl-5-phenyl-3H-benzo[e][1,4]diazepin-2-yl)-
-methyl-amine 82 (Hz141). Trimethylsilylacetylenyl analog 82
(Hz141) was obtained in 73% yield from 81 analogous to the
procedure employed above as a light yellow solid. mp:
210-211.degree. C.; IR (KBr) 3257, 3079, 2956, 2150, 1619, 1610,
1580, 1416, 1237, 880, 843 cm.sup.-1; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 0.22 (s, 9H), 2.59 (d, 3H, J=3.5 Hz), 3.56 (bs,
1H), 4.66 (bs, 1H), 6.39 (s, 1H), 7.21 (d, 1H, J=8.4 Hz), 7.39-7.65
(m, 7H); MS (EI) m/e (relative intensity) 345 (M.sup.+, 100), 344
(98), 164 (50).
[0131]
(7-Acetylenyl-4-oxy-5-phenyl-3H-benzo[e][1,4]diazepin-2-yl)-methyla-
mine 83 (Hz148). The 7-acetyleno analog 83 (Hz148) was obtained in
92% yield from 82 analogous to the procedure employed above as a
white solid. mp: 226-227.degree. C.; IR (KBr) 3275, 3245, 3075,
2102, 1618, 1599, 1580, 1467, 1416, 1333, 1235 cm.sup.-1; .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 2.65 (d, 3H, J=4.4 Hz), 2.97 (s,
1H), 3.57 (bs, 1H), 4.65 (bs, 1H), 6.20 (d, 1H, J=3.7 Hz), 7.22 (d,
1H, J=8.4 Hz), 7.42-7.58 (m, 7H). MS (EI) m/e (relative intensity)
273 (M.sup.+, 100), 272 (98).
##STR00060##
[0132] Ethyl
8-acetyleno-6-(2'-thienyl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carb-
oxylate 76 (JC221). A suspension of
7-bromo-1,3-dihydro-5-phenyl-2H-1,4-benzodiazepin-2-thione
84.sup.15 (1.6 g, 4.83 mmol), glycine (1.81 g, 24.2 mmol) and
Na.sub.2CO.sub.3 (1.84 g, 17.4 mmol) in EtOH (38 mL)-H.sub.2O (16
mL) was stirred at reflux for 5 h, poured into water (100 mL), and
then filtered to remove a small amount of
7-bromo-1,3-dihydro-5-phenyl-2H-1,4-benzodiazepin-2-one which
remained. The filtrate was extracted with CHCl.sub.3. The
CHCl.sub.3 extract was discarded; the aqueous layer was adjusted to
pH 4 with 2N HCl and then extracted with CHCl.sub.3 (3.times.25
mL). Evaporation of the CHCl.sub.3 solution gave pure acid 85 (1.2
g, 67%) as a yellow solid. Acid 85 (350 mg, 0.941 mmol) was
suspended in dry CH.sub.2Cl.sub.2 (10 mL) and DCC (223 mg, 1.08
mmol) was added. The suspension which resulted was stirred at
40.degree. C. for 2 h and then cooled to 0.degree. C. It was
filtered, and the solvent was removed in vacuum to give
8-bromo-2,4-dihydro-6-phenyl-1H-imidazo[1,2-a][1,4]benzodiazepin-1-one
3 as a brown oil. The cyclized product 86 (ca. 250 mg) was
dissolved in dry benzene (6 mL), dimethylformamide diethylacetal
(130 mg, 0.883 mmol) and triethylamine (89 mg, 0.883 mmol) were
added. The solution which resulted was stirred at room temperature
for 1 h and the solvent was removed in vacuum, The residue was then
crystallized from EtOAc-MeOH to give 87 (200 mg, 70%). A solution
of 87 (180 mg, 0.440 mmol) in dry toluene (5 mL) was treated with
1-methyl piperazine (1 mL) and heated to reflux for 5 h. The
solvent was removed in vacuum to give a gum which crystallized from
CH.sub.2Cl.sub.2-Et.sub.2O to furnish 88 (PS-1-35, 146 mg, 72%).
mp>250.degree. C.; IR (KBr) 3324, 2932, 2787, 1692, 1624, 1475,
1402, 1297, 1137, 933 cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta.
7.95 (d, 1H, J=8.8 Hz), 7.72 (dd, 1H, J=2.3 Hz, J=8.8 Hz),
7.58-7.55 (m, 2H), 7.49-7.37 (m, 4H), 7.17 (s, 1H), 5.01 (d, 1H,
J=12 Hz), 4.50-4.60 (m, 1H), 4.20-4.30 (m, 1H), 4.16 (d, 1H, J=12
Hz), 3.50-3.58 (m, 2H), 2.40-2.60 (m, 4H), 2.34 (s, 3H); MS (m/z)
465 (100).
[0133] To the suspension of compound 88 (PS-1-35, 140 mg, 0.302
mmol) in acetonitrile (4 mL) and triethylamine (3 mL) was added
bis(triphenylphosphine)-palladium (II) acetate (22.6 mg, 0.03
mmol). The solution was degassed and trimethylsilylacetylene (0.1
mL, 0.7 mmol) was added. The mixture was heated to reflux and
stirred overnight. After removal of the solvent in vacuum, the
residue was dissolved in CH.sub.2Cl.sub.2 and washed with a
saturated aqueous solution of NaHCO.sub.3 and brine. The organic
layer was dried (Na.sub.2CO.sub.3), filtered and concentrated under
vacuum. The residue was purified by flash column chromatography
(EtOAc:MeOH 9:1) to furnish the trimethylsilyl analogue 89
(PS-1-36, 100 mg, 69%) as a pale yellow solid. mp>250.degree.
C.; IR (KBr) 3436, 2936, 2794, 2154, 1682, 1625, 1489, 1136, 847
cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta. 8.0 (d, 1H, J=8.5 Hz),
7.68 (dd, 1H, J=1.9 Hz, J=8.5 Hz), 7.55-7.59 (m, 2H), 7.37-7.49 (m,
4H), 7.16 (s, 1H), 4.99 (d, 1H, J=12 Hz), 4.50-4.60 (m, 1H),
4.20-4.30 (m, 1H), 4.13 (d, 1H, J=12.4 Hz), 3.48-3.58 (m, 2H),
2.4-2.6 (m, 4H), 2.35 (s, 3H), 0.23 (s, 9H); MS (m/z) 482
(100).
[0134] A solution of the trimethylsilyl analog 89 (PS-1-36, 65 mg,
0.135 mmol) in THF (15 mL) was stirred with tetrabutylammonium
fluoride hydrate (45 mg, 0.175 mmol) at -5.degree. C. for 5 min.
After this, H.sub.2O (5 mL) was added to the solution to quench the
reaction and stirring continued at low temperature for one half
hour. The solution was extracted with EtOAc (3.times.40 mL), and
the organic layer was washed with water. After removal of the
solvent under reduced pressure, ethyl ether was added to the
residue to precipitate a solid. The mixture was filtered and the
solid was washed with CHCl.sub.3-Et.sub.2O (ca 1:15) to provide the
acetyl target 90 (PS-1-37, 40 mg, 73%). mp 223-224.degree. C.; IR
(KBr) 3298, 2935, 2786, 1695, 1628, 1364, 1136, 1002, 778
cm.sup.-1; .sup.1H NMR (CDCl.sub.3) .delta. 8.04 (d, 1H, J=8.5 Hz),
7.71 (dd, 1H, J=1.9 Hz, J=8.5 Hz), 7.55-7.58 (m, 2H), 7.36-7.48 (m,
4H), 7.17 (s, 1H), 5.0 (d, 1H, J=12.1 Hz), 4.5-4.6 (m, 1H), 4.2-4.3
(m, 1H), 4.16 (d, 1H, J=12.1 Hz), 3.5-3.6 (m, 2H), 3.08 (s, 1H),
2.4-2.6 (m, 4H), 2.35 (s, 3H); MS (m/z) (100).
##STR00061##
[0135] Ethyl
8-acetyleno-6-(2'-thienyl)-4H-benzo[1]imidazo[1,5-a][1,4]diazepine-3-carb-
oxylate 76 (JC221). The acid 27, obtained from the ester 5
(dm-I-70), was stirred with CDI in DMF, followed by stirring with
1,3-propanediol and DBU to provide 91 (DMH-D-070, the dimer of
dm-1-70). This was converted into the trimethylsilylacetylenyl
compound 92 (DMH-D-048, the dimer of XLMHe048) under standard
conditions (Pd-mediated, Heck-type coupling)..sup.4,7,8 The
bisacetylene 93 (DMH-D-053, the dimer of XHeII-053) was easily
obtained by treatment of trimethylsilyl compound 92 with fluoride
anion as shown in Scheme 18..sup.7
[0136]
8-Bromo-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxyl-
ic acid 27. The ester 5 (2 g) was dissolved in EtOH (50 mL) and aq
sodium hydroxide (10 mL, 2N) was added to the solution. The mixture
was heated to reflux for half an hour. After the EtOH was removed
under reduced pressure, the solution was allowed to cool. The pH
value was adjusted to 4 by adding 10% aq HCl dropwise. The mixture
was filtered and the solid was washed with water and ethyl ether.
The solid was dried to provide 27 (1.8 g, 96.6%): mp>250.degree.
C.; IR (KBr) 3450 (b), 2844, 1707, 1615, 1493, 1166, 700
cm.sup.-1;.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 4.14 (d, 1H,
J=12.6 Hz), 5.79 (d, 1H, 12.6 Hz), 7.41-7.54 (m, 6H), 7.88 (d, 1H,
J=8.7 Hz), 8.03 (dd, 1H, J=8.7 Hz, J=2.1 Hz), 8.47 (s, 1H); MS (EI)
m/e (rel intensity) 381 (M.sup.+, 20), 383 (19).
[0137]
1,3-Bis(8-bromo-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3--
carb-oxy) propyl diester 91 (DMH-D-070). The carboxylic acid 27 (2
g, 5.2 mmol) was dissolved in DMF (20 mL), after which CDI (1.02 g,
6.3 mmol) was added at rt and the mixture was stirred for 2 h. Then
1,3-propanediol (0.19 mL, 2.6 mmol) and DBU (0.78 mL, 5.2 mmol)
were added to the mixture and stirring continued overnight. The
reaction solution was then cooled with an ice-water bath, after
which water was added to precipitate a solid. This material was
purified further by flash chromatography on silica gel (gradient
elution, EtOAc:EtOH 20:1, 15:1, 10:1) to provide the bisbromide 91
(DMH-D-070) as a white solid (1.3 g, 61.9%): mp 187.5-189.degree.
C.; IR (KBr) 3112, 2968, 1708, 1610, 1559, 1491, 1269, 1160, 1123,
1073 cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 2.35 (m,
2H), 4.08 (d, 2H, J=12.6 Hz), 4.55 (m, 4H), 6.05 (d, 2H, J=12.6
Hz), 7.37-7.53 (m, 12H), 7.6 (d, 2H, J=2.1 Hz), 7.81 (dd, 2H, J=2.1
Hz, 8.6 Hz), 7.93 (s, 2H); .sup.13C NMR (75.5 MHz, CDCl.sub.3)
.delta. 28.2, 44.9, 61.4, 120.7, 124.2, 128.3, 129.0, 129.3, 129.6,
130.6, 134.1, 134.4, 134.7, 135.0, 138.9, 138.9, 162.6, 167.9; MS
(FAB, NBA) m/e (rel intensity) 803 (M.sup.++1, 15); Anal. Calcd.
For C.sub.39H.sub.28N.sub.6O.sub.4Br.sub.2: C, 58.23; H, 3.51; N,
10.45. Found: C, 57.92; H, 3.43; N, 10.29.
[0138]
1,3-Bis(8-trimethylsilylacetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5--
a][1,4]-diazepine-3-carboxy) propyl diester 92
(DMH-D-048)..sup.4,7,8 To a suspension of bisbromide 91 (1.005 g,
1.25 mmol) in acetonitrile (50 mL) and triethylamine (65 mL), was
added bis(triphenylphosphine)-palladium (II) acetate (0.15 g, 0.2
mmol). The solution was degassed and trimethylsilylacetylene (0.7
mL, 5 mmol) was added after which it was degassed again. The
mixture was heated to reflux and stirring maintained overnight.
After removal of the solvent under reduced pressure, the residue
was dissolved in CH.sub.2Cl.sub.2 and washed with water.
3-Mercaptopropyl functionalized silica gel (0.6 g) was added into
the organic layer and stirring continued for 1 hour. The silica
gel/Pd complex was removed by filtration and the filtrate was
concentrated under reduced pressure. The residue was purified by
flash column chromatography on silica gel (gradient elution,
EtOAc:EtOH 20:1, 15:1, 10:1) to furnish the bistrimethylsilyl dimer
92 (DMH-D-048, 680 mg, 60.8%) as a white solid: mp 169-172.degree.
C.; IR (KBr) 3449, 2950, 1725, 1720, 1715, 1496, 1250, 1160, 1080,
847 cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 0.25 (s,
18H), 2.35 (m, 2H), 4.05 (d, 2H, J=12.6 Hz), 4.55 (m, 4H), 6.02 (d,
2H, J=12.6 Hz), 7.37-7.55 (m, 14H), 7.75 (dd, 2H, J=1.8 Hz, 8.4
Hz), 7.94 (s, 2H); .sup.13C NMR (75.5 MHz, CDCl.sub.3) 8-0.3, 28.3,
44.9, 61.4, 97.4, 102.3, 122.4, 122.6, 128.0, 128.3, 129.0, 129.4,
130.5, 134.1, 134.9, 135.1, 139.0, 139.2, 139.2, 162.6, 168.5; MS
(FAB, NBA) m/e (rel intensity) 839 (M.sup.++1, 100); Anal. Calcd.
For C.sub.49H.sub.46N.sub.6O.sub.4Si.sub.2: C, 70.14; H, 5.53; N,
10.02. Found: C, 69.97; H, 5.35; N, 9.77.
1,3-Bis(8-acetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-ca-
rboxy) propyl diester 93 (DMH-D-053)..sup.7
[0139] A solution of bistrimethylsilyl dimer 92 (330 mg, 0.4 mmol)
in THF (70 mL) was stirred with tetrabutylammonium fluoride hydrate
(250 mg, 0.96 mmol) at -78.degree. C. for 5 min. After this,
H.sub.2O (35 mL) was added to the solution to quench the reaction
and stirring continued at low temperature for one half hour. The
solution was extracted with EtOAc (3.times.100 mL), and the organic
layer was washed with water. After removal of the solvent under
reduced pressure, ethyl ether was added to the residue to
precipitate a solid. The mixture was filtered and the solid was
washed with CHCl.sub.3-Et.sub.2O (ca 1:15), the bisacetylenyl dimer
93 (DMH-D-053, 220 mg, 80%) was obtained as a yellow solid: mp
172-175.degree. C.; IR (KBr) 3450, 3280, 2950, 1720, 1715, 1495,
1250, 1120, 1050 cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 2.35 (m, 2H), 3.18 (s, 2H), 4.08 (d, 2H, J=12.3 Hz), 4.56
(m, 4H), 6.04 (d, 2H, J=12.6 Hz), 7.36-7.59 (m, 14H), 7.78 (dd, 2H,
J=8.4 Hz, 1.7 Hz), 7.95 (s, 2H); .sup.13C NMR (75.5 MHz,
CDCl.sub.3) .delta. 28.8, 45.4, 61.9, 80.2, 81.3, 121.4, 122.7,
128.1, 128.3, 129.0, 129.3, 130.5, 134.2, 135.2, 135.3, 135.6,
138.9, 139.2, 162.6, 168.5; MS (FAB, NBA) m/e (rel intensity) 695
(M.sup.++1, 100).
##STR00062##
[0140] The 5-carbon linker bisbromide 94 (dm-II-26),
bis-trimethylsilylacetylenyl dimer 95 (dm-II-41) and bisacetylene
dimer 96 (dm-II-97), which are analogues of dimers DMH-D-070,
DMH-D-048 and DMH-D-053, respectively, were prepared from acid 27
under the same conditions employed for preparing dimers 91
(DMH-D-070), 92 (DMH-D-048) and 93 (DMH-D-053), respectively, by
using 1,5-pentanediol in place of 1,3-propanediol (Scheme 19).
1,5-Bis(8-bromo-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carb-ox-
y) pentyl diester 94 (dm-II-26)
[0141] A yellow powder (63.2%): mp 172-175.degree. C.; IR (KBr)
3112, 2970, 1721, 1609, 1490, 1267, 1158, 1075, 754, 697 cm.sup.-1;
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 1.62 (m, 2H), 1.90 (m,
4H), 4.07 (d, 2H, J=12.6 Hz), 4.39 (m, 4H), 6.05 (d, 2H, J=12.6
Hz), 7.37-7.53 (m, 12H), 7.58 (d, 2H, J=2.1 Hz), 7.78 (dd, 2H,
J=2.1 Hz, 8.6 Hz), 7.92 (s, 2H); .sup.13C NMR (75.5 MHz,
CDCl.sub.3) .delta. 22.5, 28.4, 44.9, 64.5, 120.7, 124.2, 128.3,
129.2, 129.3, 129.6, 130.6, 134.0, 134.5, 134.6, 135.0, 138.8,
138.9, 162.8, 167.9; MS (FAB, NBA) m/e (rel intensity) 831
(M.sup.++1, 5). Anal. Calcd. For
C.sub.41H.sub.32N.sub.6O.sub.4Br.sub.2.0.25H.sub.2O: C, 58.95; H,
3.89; N, 10.07. Found: C, 58.69; H, 3.74; N, 9.70.
1,5-Bis(8-trimethylsilylacetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]-
-diazepine-3-carboxy) pentyl diester 95 (dm-II-41)
[0142] A yellow solid (58.1%): mp 154-156.degree. C.; IR (KBr)
3426, 2955, 1727, 1720, 1612, 1495, 1251, 1174, 1076, 846
cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 0.25 (s, 18H),
1.63 (m, 2H), 1.90 (m, 4H), 4.05 (d, 2H, J=12.6 Hz), 4.39 (m, 4H),
6.03 (d, 2H, J=12.6 Hz), 7.40-7.54 (m, 14H), 7.75 (dd, 2H, J=1.8
Hz, 8.4 Hz), 7.93 (s, 2H); .sup.13C NMR (75.5 MHz, CDCl.sub.3)
8-0.3, 22.5, 28.4, 44.9, 64.5, 97.4, 102.3, 122.4, 122.6, 128.0,
128.3, 129.2, 129.4, 130.5, 134.1, 135.0, 135.1, 135.1, 138.9,
139.3, 162.8, 168.5; MS (FAB, NBA) m/e (rel intensity) 867
(M.sup.++1, 100).
[0143]
1,5-Bis(8-acetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepi-
ne-3-carboxy) pentyl diester 96 (dm-III-97). A yellow solid: mp
150-153.degree. C.; IR (KBr) 3290, 2953, 1718, 1611, 1493, 1253,
1172, 1120, 1076 cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 1.62 (m, 2H), 1.90 (m, 4H), 3.18 (s, 2H), 4.07 (d, 2H,
J=12.3 Hz), 4.38 (m, 4H), 6.04 (d, 2H, J=12.3 Hz), 7.36-7.58 (m,
14H), 7.77 (dd, 2H, J=8.4 Hz, 1.6 Hz), 7.94 (s, 2H); .sup.13C NMR
(75.5 MHz, CDCl.sub.3) .delta. 22.5, 28.4, 44.9, 64.5, 79.8, 81.3,
121.3, 122.7, 128.1, 128.3, 129.2, 129.3, 130.5, 134.1, 135.2,
135.3, 135.6, 138.8, 139.2, 162.8, 168.5; MS (FAB, NBA) m/e (rel
intensity) 723 (M.sup.++1, 13).
##STR00063##
[0144] In order to improve the water solubility of the dimers, the
oxygen-containing 5-atom linked dimers 97 (dm-III-93), 98
(dm-II-94) and 99 (dm-III-96), were designed and prepared from acid
27 under the same conditions employed for preparation of dimers
DMH-D-070, DMH-D-048 and DMH-D-053, respectively, by replacing
1,3-propanediol with diethylene glycol (Scheme 20).
Bis(8-bromo-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxy)
diethylene glycol diester 97 (dm-III-93)
[0145] A yellow solid (93.7%): mp 165-168.degree. C.; IR (KBr)
3060, 2956, 1725, 1610, 1558, 1491, 1267, 1161, 1123, 1074
cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 3.93 (t, 4H,
J=4.8 Hz), 4.06 (d, 2H, J=12.6 Hz), 4.54 (m, 4H), 6.05 (d, 2H,
J=12.6 Hz), 7.39-7.50 (m, 12H), 7.57 (d, 2H, J=2.7 Hz), 7.80 (dd,
2H, J=2.1 Hz, 8.4 Hz), 7.90 (s, 2H); .sup.13C NMR (75.5 MHz,
CDCl.sub.3) .delta. 44.9, 63.6, 69.0, 120.7, 124.2, 128.3, 129.0,
129.3, 129.6, 130.6, 134.1, 134.4, 134.6, 135.0, 138.9, 139.0,
162.5, 167.9; MS (FAB, NBA) m/e (rel intensity) 833 (M.sup.++1,
5).
Bis(8-trimethylsilylacetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diaz-
epine-3-carboxy) diethylene glycol diester 98 (dm-III-94)
[0146] A yellow solid (49.5%): mp 205-208.degree. C.; IR (KBr)
3433, 2960, 1730, 1700, 1612, 1493, 1255, 1169, 1120, 1071, 847
cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 0.25 (s, 18H),
3.93 (t, 4H, J=5.4 Hz), 4.04 (d, 2H, J=12.6 Hz), 4.55 (m, 4H), 6.04
(d, 2H, J=12.6 Hz), 7.37-7.53 (m, 14H), 7.74 (dd, 2H, J=1.2 Hz, 8.4
Hz), 7.91 (s, 2H); .sup.13C NMR (75.5 MHz, CDCl.sub.3) 8-0.3, 45.0,
63.6, 69.0, 97.5, 102.4, 122.5, 122.7, 128.1, 128.3, 129.0, 129.4,
130.5, 134.2, 135.0, 135.1, 135.2, 139.1, 139.3, 162.7, 168.6; MS
(FAB, NBA) m/e (rel intensity) 869 (M.sup.++1, 100).
Bis(8-acetylenyl-6-phenyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carb-o-
xy) diethylene glycol diester 98 (dm-III-96)
[0147] A yellow solid (81.6%): mp 173-177.degree. C.; IR (KBr)
3432, 3280, 1720, 1715, 1496, 1254, 1175, 1120, 1074 cm.sup.-1;
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 3.12 (s, 2H), 3.93 (t,
4H, J=4.5 Hz), 4.06 (d, 2H, J=12.6 Hz), 4.55 (m, 4H), 6.05 (d, 2H,
J=12.6 Hz), 7.38-7.56 (m, 14H), 7.75 (dd, 2H, J=8.4 Hz, 1.8 Hz),
7.91 (s, 2H); .sup.13C NMR (75.5 MHz, CDCl.sub.3) .delta. 45.0,
63.6, 69.0, 79.8, 81.3, 121.3, 122.7, 128.1, 128.3, 129.0, 129.3,
130.5, 134.2, 135.2, 135.3, 135.6, 139.0, 139.1, 162.6, 168.4; MS
(FAB, NBA) m/e (rel intensity) 725 (M.sup.++1, 63).
##STR00064##
[0148] The benzodiazepine 100 (bromazepam).sup.16,17 was reacted
with trimethylsilyacetylene in the presence of a palladium catalyst
to provide trimethylsilyl analog 101 (Hz157) that was methylated
with methyl iodide/sodium hydride to afford analog 102 (Hz158).
This was subjected to fluoride-mediated desilation to achieve
analog 103 (Hz160).
[0149]
7-Trimethylsilylacetylenyl-5-pyridin-2-yl-1,3-dihydro-benzo[e][1,4]-
diazepin-2-one (Hz157). Trimethylsilylacetylenyl analog 101 (Hz157)
was obtained in 76% yield from 100 analogous to the procedure
employed above as a light gray solid. mp: 242-243.degree. C.; IR
(KBr) 2956, 2155, 1690, 1616, 1492, 1332, 1248, 1018, 842, 754
cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 0.23 (s, 9H),
4.39 (s, 2H), 7.06 (d, 1H, J=8.4 Hz), 7.41 (ddd, 1H, J=7.5, 4.8,
1.2 Hz), 7.46 (d, 1H, J=1.8 Hz), 7.57 (dd, 1H, J=8.4, 1.9 Hz), 7.83
(td, 1H, J=7.7, 1.7 Hz), 7.97 (d, 1H, J=7.9 Hz), 8.41 (bs, 1H),
8.68 (d, 1H, J=4.2 Hz); MS (EI) m/e (relative intensity) 334 (35),
333 (M.sup.+, 100), 332 (57), 318 (21), 304 (31).
[0150]
7-Trimethylsilylacetylenyl-1-methyl-5-pyridin-2-yl-1,3-dihydro-benz-
o[e][1,4]diazepin-2-one (Hz158). Trimethylsilyacetylenyl analog 102
(Hz158) was obtained in 74% yield from 101 analogous to the
procedure employed above as a light grey solid. mp: 194-195.degree.
C.; IR (KBr) 2956, 2154, 1682, 1614, 1491, 1335, 1249, 881, 844,
747 cm.sup.-1; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 0.24 (s,
9H), 3.42 (s, 3H), 3.84 (d, 1H, J=10.6 Hz), 4.89 (d, 1H, J=10.6
Hz), 7.29 (d, 1H, J=7.6 Hz), 7.40 (m, 1H), 7.46 (d, 1H, J=1.9 Hz),
7.63 (dd, 1H, J=8.5, 1.9 Hz), 7.84 (td, 1H, J=7.7, 1.7 Hz), 8.09
(d, 1H, J=7.9 Hz), 8.68 (d, 1H, J=4.3 Hz); MS (EI) m/e (relative
intensity) 348 (28), 347 (M.sup.+, 100), 346 (44), 318 (34), 291
(23).
[0151]
7-Acetylenyl-1-methyl-5-pyridin-2-yl-1,3-dihydro-benzo[e][1,4]diaze-
pin-2-one (Hz160). The 7-acetyleno analog 103 (Hz160) was obtained
in 63% yield from 102 analogous to the procedure employed above as
a white solid. mp: 190-191.degree. C.; IR (KBr) 3286, 3233, 1678,
1614, 1491, 1344, 1126, 750 cm.sup.-1; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 3.07 (s, 1H), 3.86 (d, 1H, J=10.6 Hz), 4.93 (d,
1H, J=10.2 Hz), 7.32 (d, 1H, J=8.6 Hz), 7.39 (m, 1H), 7.51 (d, 1H,
J=1.8 Hz), 7.65 (dd, 1H, J=8.5, 1.9 Hz), 7.83 (td, 1H, J=7.7, 1.7
Hz), 8.11 (d, 1H, J=7.9 Hz), 8.65 (d, 1H, J=4.7 Hz); MS (EI) m/e
(relative intensity) 275 (M.sup.+, 100), 274 (35), 246 (43), 219
(30).
##STR00065##
[0152] The benzodiazepine 100 (bromazepam) was reacted with
diethylphosphorochloridate, followed by the addition of ethyl
isocyanoacetate to provide the ester 104. This was then reacted
with trimethylsilyacetylene in the presence of a palladium catalyst
to provide trimethylsilyl analog 105 (Hz165) which was subjected to
fluoride-mediated desilylation to furnish analog 106 (Hz166).
8-Trimethylsilylacetylenyl-6-pyridin-2-yl-4H-benzo[f]imidazo[1,5-a][1,4]di-
azepine-3-carboxylic acid ethyl ester 105 (Hz165)
[0153] Trimethylsilyacetylenyl analog 105 (Hz165) was obtained in
73% yield from 104 analogous to the procedure employed above as a
white solid. mp: 205-206.degree. C.; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 0.25 (s, 9H), 1.44 (t, 3H, J=7.1 Hz), 4.14 (d,
1H, J=11.0 Hz), 4.44 (m, 2H), 6.11 (d, 1H, J=10.9 Hz), 7.38 (ddd,
1H, J=7.5, 4.8, 1.1 Hz), 7.51 (s, 1H), 7.54 (d, 1H, J=8.4 Hz), 7.74
(dd, J=8.3, 1.8 Hz), 7.83 (td, 1H, J=7.7, 1.7 Hz), 7.93 (s, 1H),
8.05 (m, 1H), 8.61 (m, 1H).
[0154]
8-Acetylenyl-6-pyridin-2-yl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-
-3-carboxylic acid ethyl ester 106 (Hz166). The 7-acetyleno analog
106 (Hz166) was obtained in 98% yield from 105 analogous to the
procedure employed in above as a white solid. mp: 243-244.degree.
C.; .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 1.45 (t, 3H, J=7.1
Hz), 3.17 (s, 1H), 4.17 (d, 1H, J=10.0 Hz), 4.45 (m, 2H), 6.13 (d,
1H, J=10.4 Hz), 7.38 (ddd, 1H, J=7.5, 4.8, 1.1 Hz), 7.56 (d, 1H,
J=8.2 Hz), 7.58 (s, 1H), 7.77 (dd, 1H, J=8.6, 1.8 Hz), 7.83 (td,
1H, J=7.7, 1.8 Hz), 7.93 (s, 1H), 8.08 (m, 1H), 8.59 (m, 1H).
[0155] Some exemplary compounds falling under the scope of the
present invention are as follows:
[0156] In general, any 1,4-benzodiazepine with a 5-phenyl-like
substituent in which C(7) has been replaced with an acetylene
substituent or a trimethylsilyl acetylene substituent or any
triazolo benzodiazepine that has a corresponding substituent at
C(8) with a 6-phenyl group (alprazolam numbering system). For
example, we claim any benzodiazepine structurally related to
analogs (and other related compounds) to diazepam, alprazolam,
medazolam, and triazolam in which the C(7) or C(8) substituent has
been replaced with an acetylene or trimethylsilylacetylene
substituent.
##STR00066##
[0157] Generally, we contemplate all analogs of 1-4 above with
X'=F, Cl, Br, NO.sub.2 and/or R''=CH.sub.3, isopropyl, t-butyl,
isoxazoles. Also, all analogs of R--C.ident.C-- with R=t-butyl,
isopropyl, cyclopropyl. We believe that replacement of the halogen
atom in 1,4-benzodiazepines or the related
triazolo-1,4-benzodiazepines at C(7) or C(8) generally results in
anxiolytic activity with greatly decreased sedative/hypnotic/muscle
relaxant activity or, in some cases, no sedative hypnotic activity
compared to known agents.
##STR00067## ##STR00068## ##STR00069## ##STR00070## ##STR00071##
##STR00072## ##STR00073## ##STR00074## ##STR00075## ##STR00076##
##STR00077##
Experimental Methods
Situational Anxiety Model in Rats
[0158] Male Sprague-Dawley rats weighing 180-200 grams were
purchased from Charles River Laboratories (Wilmington, Mass.). The
rats were housed individually in suspended wire cages in a colony
room maintained at constant temperature (21.+-.2.degree. C.) and
humidity (50.+-.10%). The room was illuminated 12 hours per day
(lights on at 0600 h). The rats had ad libitum access to food and
water throughout the study. Behavioral studies were conducted
between 0600 and 1300 hours. Testing: A modification of the
Defensive Withdrawal procedure, as originally described by
Takahashi et al. (1989), was employed. Takahashi L K, Kalin N H,
Vanden Burgt J A, Sherman J E (1989) Corticotropin-releasing factor
modulates defensive-withdrawal and exploratory behavior in rats.
Behav Neurosci 103:648-654. The testing apparatus consisted of an
opaque plexiglass open field (106 cm length.times.92 cm
width.times.50 cm height), containing a cylindrical galvanized
chamber (14 cm length, 10 cm diameter) that was positioned
lengthwise against one wall, with the open end 40 cm from the
corner. The open field was illuminated by a 60 watt incandescent
bulb, and illumination was titrated by a powerstat transformer to a
23 lux reading at the entrance to the cylinder. Rats were
habituated to handling by gently stroking their dorsal surface for
approximately one minute daily for 5-6 consecutive days before
testing. To initiate testing of exploratory behavior in this
unfamiliar environment, each rat was placed within the cylinder,
which was then secured to the floor. Behavior was assessed for 15
minutes by a trained observer (unaware of treatment assignment) via
a video monitor in an adjacent room. The latency to emerge from the
cylinder, defined by the placement of all four paws into the open
field, was recorded for each rat. After testing each rat, the
plexiglass chamber and the cylinder were cleaned with 1.0% glacial
acetic acid to prevent olfactory cues from influencing the behavior
of subsequently tested rats. Drug Administration: All drugs were
administered PO 20-60 minutes prior to behavioral testing. Data
Analysis: Results were expressed as the mean.+-.1 SEM. All data
were subjected to analysis of variance (ANOVA) followed by
individual mean comparisons using Fisher's Least Significant
Difference Test where appropriate (Kirk, R. E., Experimental
Design: Procedures for the Behavioral Sciences. Brooks/Cole,
Belmont, Calif., 1968). The significance level was set at
p<0.05.
Protection from Pentylenetetrazole-Induced Seizures
[0159] Male CF1 mice weighing 20-22 g at the time of the experiment
were purchased from Charles River Laboratories (Wilmington, Mass.).
Pentylenetetrazole (PTZ) (Sigma Chemical Co, St. Louis, Mo.) was
administered at 125 mg/kg s.c. The number of animals surviving was
recorded at 30 minutes and 60 minutes after administration of
pentylenetetrazole. Drug Administration: All drugs were
administered PO 60 minutes before administration of
pentylenetetrazole. Data Analysis: The data are presented as the
percent of animals protected from death. The data were analyzed by
Chi Square statistics. The significance level was set at
p<0.05.
Protection from Electroshock-Induced Seizures
[0160] Male CF1 mice weighing 20-22 g at the time of the experiment
were purchased from Charles River Laboratories (Wilmington, Mass.).
Electroshock is administered using a Ugo Basile ECT, Unit 7801
seizure apparatus (Ugo Basile, Italy) and corneal electrodes soaked
in 0.9% saline. Mice received a shock of 30 mA for 0.3 seconds.
Drug Administration: All experimental compounds were administered
PO 60 minutes before administration of electroshock. Data Analysis:
The data are presented as the percent of animals protected from the
hind-limb extensor component of the seizure. The data were analyzed
by Chi Square statistics. The significance level was set at
p<0.05.
Open-Field Locomotor Activity in Rats
[0161] Male Sprague-Dawley rats, weighing 250-290 grams at the
beginning of the experiment were purchased from Charles River
Laboratories (Wilmington, Mass.). The animals were housed in groups
of four in a colony room maintained at constant temperature
(21.+-.2.degree. C.) and humidity (50.+-.10%). The room was
illuminated 12 hours per day (lights on at 0600 h). The rats had ad
libitum access to food and water. The testing apparatus consisted
of plexiglas chambers (42.times.42.times.30 cm) equipped with
Digiscan activity monitors (Omnitech Electronics, Columbus, Ohio)
that detect interruptions of 16 photobeams spaced 2.5 cm apart and
2.5 cm above the floor. Horizontal activity was monitored for 60
minutes. Drug Administration: All drugs were administered PO 20-60
minutes before behavioral testing. Data Analysis: Results were
expressed as the mean.+-.1 SEM. All data were subjected to analysis
of variance (ANOVA) followed by individual mean comparisons using
Fisher's Least Significant Difference Test (Kirk, 1968) where
appropriate. The significance level was set at p<0.05.
Rotorod Performance in Rats
[0162] Male Sprague-Dawley rats, weighing 180-200 grams at the
beginning of the experiment were purchased from Charles River
Laboratories (Wilmington, Mass.). The animals were housed in groups
of four in a colony room maintained at constant temperature
(21.+-.2.degree. C.) and humidity (50.+-.10%). The room was
illuminated 12 hours per day (lights on at 0600 h). The rats had ad
libitum access to food and water. The degree of muscle coordination
or balance (i.e., ataxia) was determined using a standard
accelerating rotorod treadmill (Ugo Basile, Comerio-Varese, Italy
or Columbus Instruments, Columbus, Ohio) that was 6 cm in diameter,
24 cm above the base, and run from an initial speed of 2 rpm to a
maximum speed of 20 rpm. The time each animal remained on the
rotating rod was automatically recorded, up to a maximum of 5
minutes. Each rat had three pretest acclimation trials, and the
latency from the third trial was used to counterbalance rats for
subsequent drug testing. Drug Administration: All drugs were
administered PO 20-60 minutes before behavioral testing. Data
Analysis: Results were expressed as the mean.+-.1 SEM. All data
were subjected to analysis of variance (ANOVA) followed by
individual mean comparisons using Fisher's Least Significant
Difference Test (Kirk, 1968) where appropriate. The significance
level was set at p<0.05.
Discriminative Stimulus Effects of Chlordiazepoxide in Rats
[0163] Male Sprague-Dawley rats weighing 240 to 300 g at the start
of the experiment were purchased from Charles River Laboratories
(Wilmington, Mass.). Animals were housed singly in hanging wire
cages in a room maintained at constant temperature (21-23.degree.
C.) and humidity (50.+-.10%) and illuminated 12 hours per day
(lights on at 0600 h). Throughout the study rats were restricted to
12 g of laboratory rodent chow pellets (Bio-Serv, Frenchtown, N.J.)
per day, while access to water was unlimited. All training and
testing was done Monday through Friday of each week. Twelve model
E10-10 Coulbourn operant chambers (28.times.26.times.31 cm) were
housed in light-proof, sound-attenuated, and fan-ventilated
chambers. Each operant chamber was equipped with two
non-retractable levers, requiring a downward force equivalent to 15
g (0.15 N), that were mounted 3 cm from the side wall, 3 cm above
the metal grid floor, and 5 cm from a centrally placed dipper that
delivered one 45 mg food pellet (Dustless Precision Pellets,
Bio-Serv, Frenchtown, N.J.). The experimental chambers were
connected to a Micro PDP11/73 computer using a LAB LINC interface.
A SKED-11 operating system (State System, Kalamazoo, Mich.) was
used to record and control behavior. Discrimination training: After
habituation to the operant chamber, rats were trained to alternate
daily between response levers on a Fixed Ratio 1 (FR 1) schedule of
reinforcement. Once lever pressing was well established, the
reinforcement contingency was increased incrementally to an FR 10
schedule, while maintaining the lever alternation. Next, rats were
trained to discriminate between drug (5.0 mg/kg, IP,
chlordiazepoxide) and drug vehicle (0.9% saline). Half of the rats
were randomly assigned the left lever as "drug-correct" and the
right lever as "saline-correct." The lever assignments were
reversed for the remaining animals. Every tenth response on the
drug-correct lever was reinforced on days when the rats were
pretreated with drug, whereas every tenth response on the opposite
lever was reinforced after saline injections. In each 2-week period
there were 5 drug days and 5 saline days, with the constraint that
there not be more than 3 consecutive drug or vehicle days.
Discrimination sessions were continued until each rat reached the
criterion of no more than three incorrect responses before first
food presentation in 9 out of 10 consecutive sessions. Test
sessions: Once criterion for testing was met, stimulus substitution
tests were conducted on Friday of each week. Test sessions were 10
minutes in duration. During the test sessions, the lever on which
the rat first responded 10 times resulted in reinforcement and
subsequent FR 10 reinforcement was made contingent upon pressing
this "selected" lever. The lever on which the rat first made 10
responses (the selected lever) and the total number of responses in
the session were recorded. On Monday through Thursday of each week,
training sessions were conducted to ensure that criterion for
testing was met. If any rat failed to meet the criterion for
testing, testing with that animal was postponed and discrimination
training continued until the performance criterion was attained.
Data analysis: Drug discrimination results are expressed as the
percentage of animals selecting the chlordiazepoxide-correct
lever.
Experimental Results
[0164] Table 1 (below) shows ratios of lowest effective anxiolytic
doses in the situational anxiety (SA) assay compared with lowest
effective doses producing side effects in three different models:
locomotor activity (LMA), rotorod (RR), and chlordiazepoxide-like
subjective effects as measured by the drug discrimination method
(DD).
[0165] Table 2 (below) shows effective doses in a model of epilepsy
(pentylenetetrazole-induced seizures) in mice (mg/kg, PO) for
QH-ii-066, Xli-JY-DMH, and XHe-ii-053 in comparison with diazepam,
triazolam, and DM-i-070.
Example 1
Situational Anxiety in Rats
[0166] Rats were handled daily for at least 5-6 days. They were
then placed in a dark cylinder in an illuminated open field. The
time for the rats to exit the dark cylinder was then measured.
Vehicle-treated animals remain within the dark cylinder for 10-15
minutes (total test duration is 15 minutes). This high latency to
exit the dark chamber is an index of a heightened state of anxiety.
Compounds with anxiolytic efficacy reduce latency to exit the dark
chamber. Table 1 shows that QH-ii-066, XLi-JY-DMH, and XHe-ii-053
show anxiolytic effects in the situational anxiety test at
doses>100-fold lower than doses producing sedative and ataxic
effects (see examples 2 and 3).
Example 2
Locomotor Activity in Rats
[0167] Rats were placed in an open field and the total distance
covered by the rat was measured. The test duration was 60 minutes.
Compounds producing sedative effects decrease the distance covered.
Table 1 shows that QH-ii-066, XLi-JY-DMH, and XHe-ii-053 are less
effective in producing sedative or hypnotic effects than diazepam
or triazolam.
Example 3
Rotorod Performance in Rats
[0168] Rats were placed on a slowly rotating rod and the speed of
rotation was gradually increased. The time on the rod for each rat
was recorded. Compounds producing ataxia (motor incoordination)
decrease the time spent on the rod compared with vehicle-treated
animals. Table 1 shows that QH-ii-066, XLi-JY-DMH, and XHe-ii-053
are less potent in producing ataxia than diazepam or triazolam.
Thus, they are likely better drugs clinically because they have
decreased side effects [decreased sedation (example 2) and ataxia
(example 3)].
Example 4
Drug Discrimination in Rats
[0169] Animals are taught to emit one response if they just
received drug and a different response if they just received
saline. The animals learn to discriminate between a "drug state"
and a "no drug state". The rats were trained to discriminate
between a state induced by a typical benzodiazepine
chlordiazepoxide (CDP; "drug state") and a state induced by vehicle
(methocel: "no drug state"). Table 1 shows that QH-ii-066,
XLi-JY-DMH, and XHe-ii-053 are less potent in producing CDP-like
effects than diazepam or triazolam and thus may have reduced abuse
potential compared with CDP.
Example 5
Seizure Protection in Mice
[0170] Mice treated with certain compounds of the present invention
were subjected to pentylenetetrazole (PTZ) at 125 mg/kg to induce
seizures. The percent of animals protected from death within one
hour of PTZ was measured. Table 2 shows that QH-ii-066 and
XLi-JY-DMH have anticonvulsant effects against PTZ-induced seizures
at doses comparable to those for diazepam and triazolam. Table 2
also shows that XHe-ii-053 is effective against PTZ-induced
seizures.
TABLE-US-00001 TABLE 1 Antianxiety/ Antianxiety/ Antianxiety/
sedation ataxia abuse liability Diazepam 10 100 5 QH-ii-066 100
>100 30 Triazolam 300 100 30 XLi-JY-DMH 10000 10000 1000
DM-i-070 >100 >100 10 XHe-ii-053 >300 >300 >300
TABLE-US-00002 TABLE 2 PTZ Seizures (mg/kg, PO Diazepam <10
QH-ii-066 <30 Triazolam <1.0 XLi-JY-DMH <1.0 DM-i-070
<100 XHe-ii-053 .ltoreq.100
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