U.S. patent application number 11/470695 was filed with the patent office on 2007-02-01 for bicyclic pyrazolyl and imidazolyl compounds and uses thereof.
This patent application is currently assigned to Pfizer Inc. Invention is credited to Philip A. Carpino, Robert L. Dow, David A. Griffith.
Application Number | 20070027133 11/470695 |
Document ID | / |
Family ID | 34572986 |
Filed Date | 2007-02-01 |
United States Patent
Application |
20070027133 |
Kind Code |
A1 |
Carpino; Philip A. ; et
al. |
February 1, 2007 |
BICYCLIC PYRAZOLYL AND IMIDAZOLYL COMPOUNDS AND USES THEREOF
Abstract
Compounds of Formula (I) are described herein. ##STR1## The
compounds have been shown to act as cannabinoid receptor ligands
and are therefore useful in the treatment of diseases linked to the
mediation of the cannabinoid receptors in animals.
Inventors: |
Carpino; Philip A.; (Groton,
CT) ; Dow; Robert L.; (Waterford, CT) ;
Griffith; David A.; (Old Saybrook, CT) |
Correspondence
Address: |
PFIZER INC.
PATENT DEPARTMENT, MS8260-1611
EASTERN POINT ROAD
GROTON
CT
06340
US
|
Assignee: |
Pfizer Inc
|
Family ID: |
34572986 |
Appl. No.: |
11/470695 |
Filed: |
September 7, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10971599 |
Oct 22, 2004 |
7151097 |
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11470695 |
Sep 7, 2006 |
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60518280 |
Nov 7, 2003 |
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Current U.S.
Class: |
514/211.05 ;
514/212.06; 514/221; 540/490; 540/502; 540/521 |
Current CPC
Class: |
A61P 25/00 20180101;
A61P 25/06 20180101; A61P 25/14 20180101; A61P 25/32 20180101; A61P
15/00 20180101; A61P 29/00 20180101; A61P 1/00 20180101; A61P 25/08
20180101; A61P 7/02 20180101; A61P 1/04 20180101; A61P 25/22
20180101; A61P 15/10 20180101; A61P 25/30 20180101; C07D 513/04
20130101; A61P 25/18 20180101; A61P 25/16 20180101; A61P 25/36
20180101; C07D 487/14 20130101; A61P 25/24 20180101; C07D 487/04
20130101; A61P 25/10 20180101; A61P 3/04 20180101; C07D 471/04
20130101; A61P 43/00 20180101; A61P 25/34 20180101; A61P 1/14
20180101; A61P 3/10 20180101; A61P 25/20 20180101; A61P 11/00
20180101; A61P 25/28 20180101 |
Class at
Publication: |
514/211.05 ;
514/212.06; 514/221; 540/490; 540/502; 540/521 |
International
Class: |
A61K 31/554 20070101
A61K031/554; A61K 31/553 20070101 A61K031/553; A61K 31/5513
20070101 A61K031/5513; A61K 31/55 20070101 A61K031/55; C07D 487/02
20070101 C07D487/02; C07D 491/02 20070101 C07D491/02; C07D 498/02
20070101 C07D498/02 |
Claims
1-181. (canceled)
182. A pharmaceutical composition comprising (1) a compound of
Formula (I) ##STR60## wherein A is nitrogen and B is carbon, or A
is carbon and B is nitrogen; R.sup.0 is an aryl optionally
substituted with one or more substituents, or a heteroaryl
optionally substituted with one or more substituents; R.sup.1 is
aryl optionally substituted with one or more substituents,
heteroaryl optionally substituted with one or more substituents,
--CH.dbd.CH--R.sup.1a, or --CH.sub.2CH.sub.2--R.sup.1a, where
R.sup.1a is hydrogen or a chemical moiety selected from
(C.sub.1-C.sub.8)alkyl, 3- to 8-membered partially or fully
saturated carbocyclic ring(s), 3- to 6-membered partially or fully
saturated heterocycle, aryl, or heteroaryl, where the chemical
moiety is optionally substituted with one or more substituents; X
is O, S, SO, SO.sub.2, --N(R.sup.2a)-- or
--C(R.sup.2b)(R.sup.2c)--, where R.sup.2a, R.sup.2b and R.sup.2c
are each independently hydrogen, (C.sub.1-C.sub.4)alkyl,
halo-substituted (C.sub.1-C.sub.4)alkyl or (C.sub.1-C.sub.5)acyl;
R.sup.3a and R.sup.3b are each independently hydrogen,
(C.sub.1-C.sub.6)alkyl, or halo-substituted C.sub.1-C.sub.6)alkyl,
or either R.sup.3a or R.sup.3b taken together with R.sup.4 forms a
fully or partially saturated 5- to 6-membered heterocyclic ring,
where the heterocyclic ring optionally contains an additional
heteroatom selected from oxygen, nitrogen or sulfur and is
optionally substituted with one or more substituents; and R.sup.4
is a chemical moiety selected from the group consisting of
(C.sub.1-C.sub.8)alkyl, aryl, heteroaryl,
aryl(C.sub.1-C.sub.4)alkyl, a 3- to 8-membered partially or fully
saturated carbocyclic ring(s), heteroaryl(C.sub.1-C.sub.3)alkyl,
5-6 membered lactone, 5- to 6-membered lactam, and a 3- to
8-membered partially or fully saturated heterocycle, where said
chemical moiety is optionally substituted with one or more
substituents, or R.sup.4 taken together with either R.sup.3a or
R.sup.3b forms a fully or partially saturated 5- to 6-membered
heterocyclic ring, where the heterocyclic ring optionally contains
an additional heteroatom selected from oxygen, nitrogen or sulfur
and is optionally substituted with one or more substituents; a
pharmaceutically acceptable salt thereof, or a solvate or hydrate
of said compound or said salt; (2) a pharmaceutically acceptable
excipient, diluent, or carrier; and (3) at least one additional
pharmaceutical agent.
183. The composition of claim 182 wherein said additional
pharmaceutical agent is a nicotine receptor partial agonist, an
opioid antagonist, a dopaminergic agent, an attention deficit
disorder agent, or an anti-obesity agent.
184. The composition of claim 183 wherein said anti-obesity agent
is selected from the group consisting of an apo-B/MTP inhibitor, a
11.beta.-hydroxy steroid dehydrogenase-1 inhibitor, peptide
YY.sub.3-36 or an analog thereof, a MCR-4 agonist, a CCK-A agonist,
a monoamine reuptake inhibitor, a sympathomimetic agent, a
.beta..sub.3 adrenergic receptor agonist, a dopamine agonist, a
melanocyte-stimulating hormone receptor analog, a 5-HT2c receptor
agonist, a melanin concentrating hormone antagonist, leptin, a
leptin analog, a leptin receptor agonist, a galanin antagonist, a
lipase inhibitor, a bombesin agonist, a neuropeptide-Y receptor
antagonist, a thyromimetic agent, dehydroepiandrosterone or analog
thereof, a glucocorticoid receptor antagonist, an orexin receptor
antagonist, a glucagon-like peptide-1 receptor agonist, a ciliary
neurotrophic factor, a human agouti-related protein antagonist, a
ghrelin receptor antagonist, a histamine 3 receptor antagonist or
inverse agonist, and a neuromedin U receptor agonist.
185. A method for treating a disease, condition or disorder which
is modulated by a cannabinoid receptor antagonist in animals
comprising the step of administering to an animal in need of such
treatment a therapeutically effective amount of a compound of
Formula (I) ##STR61## where A is nitrogen and B is carbon, or A is
carbon and B is nitrogen; R.sup.0 is an aryl optionally substituted
with one or more substituents, or a heteroaryl optionally
substituted with one or more substituents; R.sup.1 is aryl
optionally substituted with one or more substituents, heteroaryl
optionally substituted with one or more substituents,
--CH.dbd.CH--R.sup.1a, or CH.sub.2CH.sub.2--R.sup.1a, where
R.sup.1a is hydrogen or a chemical moiety selected from
(C.sub.1-C.sub.8)alkyl, 3- to 8-membered partially or fully
saturated carbocyclic ring(s), 3- to 6-membered partially or fully
saturated heterocycle, aryl, or heteroaryl, where the chemical
moiety is optionally substituted with one or more substituents; X
is O, S, SO, SO.sub.2, --N(R.sup.2a)-- or
--C(R.sup.2b)(R.sup.2c)--, where R.sup.2a, R.sup.2b and R.sup.2c
are each independently hydrogen, (C.sub.1-C.sub.4)alkyl,
halo-substituted (C.sub.1-C.sub.4)alkyl or (C.sub.1-C.sub.5)acyl;
R.sup.3a and R.sup.3b are each independently hydrogen,
(C.sub.1-C.sub.6)alkyl, or halo-substituted (C.sub.1-C.sub.6)alkyl,
or either R.sup.3a or R.sup.3b taken together with R.sup.4 forms a
fully or partially saturated 5- to 6-membered heterocyclic ring,
where the heterocyclic ring optionally contains an additional
heteroatom selected from oxygen, nitrogen or sulfur and is
optionally substituted with one or more substituents; and R.sup.4
is a chemical moiety selected from the group consisting of
(C.sub.1-C.sub.8)alkyl, aryl, heteroaryl,
aryl(C.sub.1-C.sub.4)alkyl, a 3- to 8-membered partially or fully
saturated carbocyclic ring(s), heteroaryl(C.sub.1-C.sub.3)alkyl,
5-6 membered lactone, 5- to 6-membered lactam, and a 3- to
8-membered partially or fully saturated heterocycle, where said
chemical moiety is optionally substituted with one or more
substituents, or R.sup.4 taken together with either R.sup.3a or
R.sup.3b forms a fully or partially saturated 5- to 6-membered
heterocyclic ring, where the heterocyclic ring optionally contains
an additional heteroatom selected from oxygen, nitrogen or sulfur
and is optionally substituted with one or more substituents; a
pharmaceutically acceptable salt thereof, or a solvate or hydrate
of said compound or said salt, wherein said compound is
administered in combination with a nicotine receptor partial
agonist, an opioid antagonist, a dopaminergic agent, an attention
deficit disorder agent, or an anti-obesity agent.
186. (canceled)
187. The method of claim 186 wherein said anti-obesity agent is
selected from the group consisting of an apo-B/MTP inhibitor, a
11.beta.-hydroxy steroid dehydrogenase-1 inhibitor, peptide
YY.sub.3-36 or an analog thereof, a MCR-4 agonist, a CCK-A agonist,
a monoamine reuptake inhibitor, a sympathomimetic agent, a 3
adrenergic receptor agonist, a dopamine agonist, a
melanocyte-stimulating hormone receptor analog, a 5-HT2c receptor
agonist, a melanin concentrating hormone antagonist, leptin, a
leptin analog, a leptin receptor agonist, a galanin antagonist, a
lipase inhibitor, a bombesin agonist, a neuropeptide-Y receptor
antagonist, a thyromimetic agent, dehydroepiandrosterone or analog
thereof, a glucocorticoid receptor antagonist, an orexin receptor
antagonist, a glucagon-like peptide-1 receptor agonist, a ciliary
neurotrophic factor, a human agouti-related protein antagonist, a
ghrelin receptor antagonist, a histamine 3 receptor antagonist or
inverse agonist, and a neuromedin U receptor agonist.
188-189. (canceled)
190. A method for treating a disease, condition or disorder
modulated by a cannabinoid receptor antagonist comprising the step
of administering a pharmaceutical composition of claim 182.
191. (canceled)
192. The method of claim 190 wherein said additional pharmaceutical
agent is a nicotine partial agonist, an opioid antagonist, a
dopaminergic agent, an attention deficit disorder agent, or an
anti-obesity agent.
193. The method of claim 192 wherein said anti-obesity agent is
selected from the group consisting of an apo-B/MTP inhibitor, a
11.beta.-hydroxy steroid dehydrogenase-1 inhibitor, peptide
YY.sub.3-36 or an analog thereof, a MCR-4 agonist, a CCK-A agonist,
a monoamine reuptake inhibitor, a sympathomimetic agent, a
.beta..sub.3 adrenergic receptor agonist, a dopamine agonist, a
melanocyte-stimulating hormone receptor analog, a 5-HT2c receptor
agonist, a melanin concentrating hormone antagonist, leptin, a
leptin analog, a leptin receptor agonist, a galanin antagonist, a
lipase inhibitor, a bombesin agonist, a neuropeptide-Y receptor
antagonist, a thyromimetic agent, dehydroepiandrosterone or analog
thereof, a glucocorticoid receptor antagonist, an orexin receptor
antagonist, a glucagon-like peptide-1 receptor agonist, a ciliary
neurotrophic factor, a human agouti-related protein antagonist, a
ghrelin receptor antagonist, a histamine 3 receptor antagonist or
inverse agonist, and a neuromedin U receptor agonist.
194. The method of claim 190, 192 or 193 wherein said disease,
condition or disorder modulated by a cannabinoid receptor
antagonist is obesity, bulimia, attention deficit disorder,
Parkinson's disease, dementia, alcoholism, or tobacco abuse.
195. A method for treating a disease, condition or disorder
modulated by a cannabinoid receptor antagonist in animals
comprising the step of administering to an animal in need of such
treatment two separate pharmaceutical compositions comprising (i) a
first composition comprising a compound of Formula (I) ##STR62##
wherein A is nitrogen and B is carbon, or A is carbon and B is
nitrogen; R.sup.0 is an aryl optionally substituted with one or
more substituents, or a heteroaryl optionally substituted with one
or more substituents; R.sup.1 is aryl optionally substituted with
one or more substituents, heteroaryl optionally substituted with
one or more substituents, --CH.dbd.CH--R.sup.1a, or
--CH.sub.2CH.sub.2--R.sup.1a, where R.sup.1a is hydrogen or a
chemical moiety selected from (C.sub.1-C.sub.8)alkyl, 3- to
8-membered partially or fully saturated carbocyclic ring(s), 3- to
6-membered partially or fully saturated heterocycle, aryl, or
heteroaryl, where the chemical moiety is optionally substituted
with one or more substituents; X is O, S, SO, SO.sub.2,
--N(R.sup.2a)-- or --C(R.sup.2b)(R.sup.2c)--, where R.sup.2a,
R.sup.2b and R.sup.2c are each independently hydrogen,
(C.sub.1-C.sub.4)alkyl, halo-substituted (C.sub.1-C.sub.4)alkyl or
(C.sub.1-C.sub.5 acyl; R.sup.3a and R.sup.3b are each independently
hydrogen, (C.sub.1-C.sub.6)alkyl, or halo-substituted or either
R.sup.3a or R.sup.3b taken together with R.sup.4 forms a fully or
partially saturated 5- to 6-membered heterocyclic ring, where the
heterocyclic ring optionally contains an additional heteroatom
selected from oxygen, nitrogen or sulfur and is optionally
substituted with one or more substituents; and R.sup.4 is a
chemical moiety selected from the group consisting of
(C.sub.1-8)alkyl, aryl, heteroaryl, aryl(C.sub.1-C.sub.4)alkyl, a
3- to 8-membered partially or fully saturated carbocyclic ring(s),
heteroaryl(C.sub.1-C.sub.3)alkyl, 5-6 membered lactone, 5- to
6-membered lactam, and a 3- to 8-membered partially or fully
saturated heterocycle, where said chemical moiety is optionally
substituted with one or more substituents, or R.sup.4 taken
together with either R.sup.3a or R.sup.3b forms a fully or
partially saturated 5- to 6-membered heterocyclic ring, where the
heterocyclic ring optionally contains an additional heteroatom
selected from oxygen, nitrogen or sulfur and is optionally
substituted with one or more substituents; or a pharmaceutically
acceptable salt thereof or a solvate or hydrate of said compound or
said salt, and a pharmaceutically acceptable excipient, diluent, or
carrier, and (ii) a second composition comprising at least one
additional pharmaceutical agent and a pharmaceutically acceptable
excipient, diluent, or carrier.
196. The method of claim 195 wherein said at least one additional
pharmaceutical agent is a nicotine partial agonist, an opioid
antagonist, a dopaminergic agent, an attention deficit disorder
agent, or an anti-obesity agent.
197. The method of claim 196 wherein said anti-obesity agent is
selected from the group consisting of an apo-B/MTP inhibitor, a
11.beta.-hydroxy steroid dehydrogenase-1 inhibitor, peptide
YY.sub.3-36 or an analog thereof, a MCR-4 agonist, a CCK-A agonist,
a monoamine reuptake inhibitor, a sympathomimetic agent, a
.beta..sub.3 adrenergic receptor agonist, a dopamine agonist, a
melanocyte-stimulating hormone receptor analog, a 5-HT2c receptor
agonist, a melanin concentrating hormone antagonist, leptin, a
leptin analog, a leptin receptor agonist, a galanin antagonist, a
lipase inhibitor, a bombesin agonist, a neuropeptide-Y receptor
antagonist, a thyromimetic agent, dehydroepiandrosterone or analog
thereof a glucocorticoid receptor antagonist, an orexin receptor
antagonist, a glucagon-like peptide-1 receptor agonist, a ciliary
neurotrophic factor, a human agouti-related protein antagonist, a
ghrelin receptor antagonist, a histamine 3 receptor antagonist or
inverse agonist, and a neuromedin U receptor agonist.
198. The method of claim 195 wherein said first composition and
said second composition are administered simultaneously.
199. The method of claim 195 wherein said first composition and
said second composition are administered sequentially and in any
order.
200. The composition of claim 182 wherein said additional
pharmaceutical agent is a cognitive impairment agent.
201. The composition of claim 200 wherein said cognitive impairment
agent is donepezil hydrochloride.
Description
[0001] This patent application claims the benefit of U.S.
Provisional Patent Application Ser. No. 60/518,280 filed on Nov. 7,
2003 and incorporated herein by reference in its entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to bicyclic pyrazolyl and
imidazolyl compounds. The compounds have been found to be
cannabinoid receptor ligands, in particular CB1 receptor
antagonists, and are therefore useful for treating diseases,
conditions and/or disorders modulated by cannabinoid receptor
antagonists.
BACKGROUND
[0003] Obesity is a major public health concern because of its
increasing prevalence and associated health risks. Obesity and
overweight are generally defined by body mass index (BMI), which is
correlated with total body fat and estimates the relative risk of
disease. BMI is calculated by weight in kilograms divided by height
in meters squared (kg/m.sup.2). Overweight is typically defined as
a BMI of 25-29.9 kg/m.sup.2, and obesity is typically defined as a
BMI of 30 kg/m.sup.2. See, e.g., National Heart, Lung, and Blood
Institute, Clinical Guidelines on the Identification, Evaluation,
and Treatment of Overweight and Obesity in Adults, The Evidence
Report, Washington, D.C.: U.S. Department of Health and Human
Services, NIH publication no. 984083 (1998).
[0004] The increase in obesity is of concern because of the
excessive health risks associated with obesity, including coronary
heart disease, strokes, hypertension, type 2 diabetes mellitus,
dyslipidemia, sleep apnea, osteoarthritis, gall bladder disease,
depression, and certain forms of cancer (e.g., endometrial, breast,
prostate, and colon). The negative health consequences of obesity
make it the second leading cause of preventable death in the United
States and impart a significant economic and psychosocial effect on
society. See, McGinnis M, Foege W H., "Actual Causes of Death in
the United States," JAMA, 270, 2207-12 (1993).
[0005] Obesity is now recognized as a chronic disease that requires
treatment to reduce its associated health risks. Although weight
loss is an important treatment outcome, one of the main goals of
obesity management is to improve cardiovascular and metabolic
values to reduce obesity-related morbidity and mortality. It has
been shown that 5-10% loss of body weight can substantially improve
metabolic values, such as blood glucose, blood pressure, and lipid
concentrations. Hence, it is believed that a 5-10% intentional
reduction in body weight may reduce morbidity and mortality.
[0006] Currently available prescription drugs for managing obesity
generally reduce weight by inducing satiety or decreasing dietary
fat absorption. Satiety is achieved by increasing synaptic levels
of norepinephrine, serotonin, or both. For example, stimulation of
serotonin receptor subtypes 1B, 1D, and 2C and 1- and 2-adrenergic
receptors decreases food intake by regulating satiety. See, Bray G
A, "The New Era of Drug Treatment. Pharmacologic Treatment of
Obesity: Symposium Overview," Obes Res., 3(suppl 4), 415s-7s
(1995). Adrenergic agents (e.g., diethylpropion, benzphetamine,
phendimetrazine, mazindol, and phentermine) act by modulating
central norepinephrine and dopamine receptors through the promotion
of catecholamine release. Older adrenergic weight-loss drugs (e.g.,
amphetamine, methamphetamine, and phenmetrazine), which strongly
engage in dopamine pathways, are no longer recommended because of
the risk of their abuse. Fenfluramine and dexfenfluramine, both
serotonergic agents used to regulate appetite, are no longer
available for use.
[0007] More recently, CB1 cannabinoid receptor antagonists/inverse
agonists have been suggested as potential appetite suppressants.
See, e.g., Amone, M., et al., "Selective Inhibition of Sucrose and
Ethanol Intake by SR141716, an Antagonist of Central Cannabinoid
(CB1) Receptors," Psychopharmacol, 132, 104-106 (1997); Colombo,
G., et al., "Appetite Suppression and Weight Loss after the
Cannabinoid Antagonist SR141716," Life Sci., 63, PL113-PL117
(1998); Simiand, J., et al., "SR141716, a CB1 Cannabinoid Receptor
Antagonist, Selectively Reduces Sweet Food Intake in Marmose,"
Behav. Pharmacol., 9, 179-181 (1998); and Chaperon, F., et al.,
"Involvement of Central Cannabinoid (CB1) Receptors in the
Establishment of Place Conditioning in Rats," Psychopharmacology,
135, 324-332 (1998). For a review of cannabinoid CB1 and CB2
receptor modulators, see Pertwee, R. G., "Cannabinoid Receptor
Ligands: Clinical and Neuropharmacological Considerations, Relevant
to Future Drug Discovery and Development," Exp. Opin. Invest.
Drugs, 9(7), 1553-1571 (2000).
[0008] Although investigations are on-going, there still exists a
need for a more effective and safe therapeutic treatment for
reducing or preventing weight-gain.
[0009] In addition to obesity, there also exists an unmet need for
treatment of alcohol abuse. Alcoholism affects approximately 10.9
million men and 4.4 million women in the United States.
Approximately 100,000 deaths per year have been attributed to
alcohol abuse or dependence. Health risks associated with
alcoholism include impaired motor control and decision making,
cancer, liver disease, birth defects, heart disease, drug/drug
interactions, pancreatitis and interpersonal problems. Studies have
suggested that endogenous cannabinoid tone plays a critical role in
the control of ethanol intake. The endogenous CB1 receptor
antagonist SR-141716A has been shown to block voluntary ethanol
intake in rats and mice. See, Amone, M., et al., "Selective
Inhibition of Sucrose and Ethanol Intake by SR141716, an Antagonist
of Central Cannabinoid (CB1) Receptors," Psychopharmacol, 132,
104-106 (1997). For a review, see Hungund, B. L and B. S.
Basavarajappa, "Are Anadamide and Cannabinoid Receptors involved in
Ethanol Tolerance? A Review of the Evidence," Alcohol &
Alcoholism. 35(2) 126-133, 2000.
[0010] Current treatments for alcohol abuse or dependence generally
suffer from non-compliance or potential hepatotoxicity; therefore,
there is a high unmet need for more effective treatment of alcohol
abuse/dependence.
SUMMARY
[0011] The present invention provides compounds of Formula (I):
##STR2## wherein
[0012] A is nitrogen and B is carbon, or A is carbon and B is
nitrogen;
[0013] R.sup.0 is an aryl optionally substituted with one or more
substituents, or a heteroaryl optionally substituted with one or
more substituents;
[0014] R.sup.1 is aryl optionally substituted with one or more
substituents, heteroaryl optionally substituted with one or more
substituents, --CH.dbd.CH--R.sup.1a, or
--CH.sub.2CH.sub.2--R.sup.1a, where R.sup.1a is hydrogen or a
chemical moiety selected from (C.sub.1-C.sub.8)alkyl, 3- to
8-membered partially or fully saturated carbocyclic ring(s), 3- to
6-membered partially or fully saturated heterocycle, aryl,
heteroaryl, where the chemical moiety is optionally substituted
with one or more substituents;
[0015] X is O, S, SO, SO.sub.2, --N(R.sup.2a)-- or
--C(R.sup.2b)(R.sup.2c)--, where R.sup.2a, R.sup.2b and R.sup.2c
are each independently hydrogen, (C.sub.1-C.sub.4)alkyl,
halo-substituted (C.sub.1-C.sub.4)alkyl or (C.sub.1-C.sub.5)acyl;
(preferably, R.sup.2a is hydrogen, (C.sub.1-C.sub.4)alkyl, or
fluoro-substituted (C.sub.1-C.sub.4)alkyl); and at least one of
R.sup.2b and R.sup.2c is (C.sub.1-C.sub.4)alkyl or
fluoro-substituted (C.sub.1-C.sub.4)alkyl, or both R.sup.2b and
R.sup.2c are hydrogen);
[0016] R.sup.3a and R.sup.3b are each independently hydrogen,
(C.sub.1-C.sub.6)alkyl, or halo-substituted (C.sub.1-C.sub.6)alkyl
(preferably, R.sup.3a and R.sup.3b are each independently hydrogen,
(C.sub.1-C.sub.4)alkyl, or fluoro-substituted
(C.sub.1-C.sub.4)alkyl), or either R.sup.3a or R.sup.3b taken
together with R.sup.4 forms a fully or partially saturated 5- to
6-membered heterocyclic ring, where the heterocyclic ring
optionally contains an additional heteroatom selected from oxygen,
nitrogen or sulfur and is optionally substituted with one or more
substituents; and
[0017] R.sup.4 is a chemical moiety selected from the group
consisting of (C.sub.1-C.sub.8)alkyl, aryl, heteroaryl,
aryl(C.sub.1-C.sub.4)alkyl, a 3- to 8-membered partially or fully
saturated carbocyclic ring(s), heteroaryl(C.sub.1-C.sub.3)alkyl,
5-6 membered lactone, 5- to 6-membered lactam, and a 3- to
8-membered partially or fully saturated heterocycle, where said
chemical moiety is optionally substituted with one or more
substituents, or R.sup.4 taken together with either R.sup.3a or
R.sup.3b forms a fully or partially saturated 5- to 6-membered
heterocyclic ring, where the heterocyclic ring optionally contains
an additional heteroatom selected from oxygen, nitrogen or sulfur
and is optionally substituted with one or more substituents;
[0018] a pharmaceutically acceptable salt thereof, a prodrug of the
compound or the salt, or a solvate or hydrate of the compound, the
salt or the prodrug.
[0019] Preferably, R.sup.0 and R.sup.1 are each independently a
substituted phenyl. More preferably, R.sup.0 and R.sup.1 are each
independently a phenyl substituted with one to three substituents
independently selected from the group consisting of halo
(preferably, chloro or fluoro), (C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkyl, halo-substituted (C.sub.1-C.sub.4)alkyl
(preferably fluoro-substituted alkyl, more preferably,
trifluoromethyl) and cyano. Most preferably, R.sup.0 is
2-chlorophenyl, 2-fluorophenyl, 2,4-dichlorophenyl,
2-fluoro-4-chlorophenyl, 2-chloro-4-fluorophenyl, 2-methylphenyl or
2,4-difluorophenyl; and R.sup.1 is 4-chlorophenyl, 4-cyanophenyl,
4-ethylphenyl, 4-isopropylphenyl, 4-ethoxyphenyl,
4-isopropoxyphenyl, 4-trifluoromethylphenyl, or 4-fluorophenyl.
[0020] Preferably, R.sup.4 is a chemical moiety selected from the
group consisting of (C.sub.1-C.sub.8)alkyl,
aryl(C.sub.1-C.sub.4)alkyl, 3- to 8-membered partially or fully
saturated carbocyclic ring(s), and 3- to 8-membered partially or
fully saturated heterocycle, where said chemical moiety is
optionally substituted with one or more substituents or R.sup.4
taken together with either R.sup.3a or R.sup.3b forms a fully or
partially saturated 5- to 6-membered heterocyclic ring, where said
heterocyclic ring is optionally substituted with one or more
substituents. More preferably, R.sup.4 is (C.sub.1-C.sub.8)alkyl,
halo-substituted (C.sub.1-C.sub.8)alkyl (preferably,
fluoro-substituted (C.sub.1-C.sub.8)alkyl), cyclopentyl,
cyclohexyl, piperidin-1-yl, pyrrolidin-1-yl, or morpholin-1-yl.
[0021] In a preferred embodiment of the present invention, a
compound of Formula (II) is provided: ##STR3## wherein
[0022] A, B, X, R.sup.2a, R.sup.2b, R.sup.2c, R.sup.3a, R.sup.3b,
and R.sup.4 are as defined above (preferred groups are also defined
above);
[0023] R.sup.0a, R.sup.0b, R.sup.1b, and R.sup.1c are each
independently halo, (C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkyl, halo-substituted (C.sub.1-C.sub.4)alkyl, or
cyano (preferably, R.sup.0a is chloro, fluoro, or methyl; R.sup.0b
is chloro, fluoro or hydrogen (i.e., m is 0); R.sup.1c is chloro,
fluoro, (C.sub.1-C.sub.4)alkyl, trifluoromethyl,
(C.sub.1-C.sub.4)alkoxy, or cyano; and R.sup.1b is hydrogen (i.e.,
n is 0));
[0024] n and m are each independently 0, 1 or 2;
[0025] a pharmaceutically acceptable salt thereof, a prodrug of the
compound or the salt, or a solvate or hydrate of the compound, the
salt or the prodrug.
[0026] Preferred compounds of the present invention include:
3-(4-chlorophenyl)-2-(2-chlorophenyl)-7-(2,2,2-trifluoroethyl)-6,7-dihydr-
o-2H,5H-4-oxa-1,2,7-triaza-azulen-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-7-(2,2-difluoropropyl)-6,7-dihydro--
2H,5H-4-oxa-1,2,7-triaza-azulen-8-one;
3-(4-chloro-phenyl)-2-(2-chloro-phenyl)-7-(2,2-difluorobutyl)-6,7-dihydro-
-2H,5H-4-oxa-1,2,7-triaza-azulen-8-one;
3-(4-chloro-phenyl)-2-(2-chloro-phenyl)-7-isopropyl-6,7-dihydro-2H,5H-4-o-
xa-1,2,7-triaza-azulen-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-7-(2,2,2-trifluoroethyl)-4,5,6,7-te-
trahydro-2H-pyrazolo[4,3-e][1,4]diazepin-8-one;
1-(4-chlorophenyl)-2-(2-chlorophenyl)-5,6,7,7a,8,9-hexahydro-2H-2,3,4a,9--
tetraazacyclopenta[f]azulen-4-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-7-(2,2,2-trifluoroethyl)-4,5,6,7-te-
trahydro-2H-pyrazolo[4,3-e][1,4]diazepin-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-6,6-dimethyl-7-(2,2,2-trifluoroethy-
l)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-e][1,4]diazepin-8-one;
2-(2-chlorophenyl)-1-(4-chlorophenyl)-2,5,6,7,8,8a,9,10-octahydro-2,3,4a,-
10-tetraazabenzo[f]azulen-4-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4,6,6-trimethyl-7-(2,2,2-trifluoroe-
thyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-e][1,4]diazepin-8-one;
2-(2-chlorophenyl)-1-(4-chlorophenyl)-9-methyl-5,6,7,7a,8,9-hexahydro-2H--
2,3,4a,9-tetraazacyclopenta[f]azulen-4-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4-methyl-7-(2,2,2-trifluoroethyl)-4-
,5,6,7-tetrahydro-2H-pyrazolo[4,3-e][1,4]diazepin-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-7-(2,2,2-trifluoroethyl)-6,7-dihydr-
o-2H,5H-4-thia-1,2,7-triaza-azulen-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4-oxo-7-(2,2,2-trifluoroethyl)-4,5,-
6,7-tetrahydro-2H-4.lamda..sup.4-thia-1,2,7-triaza-azulen-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4,4-dioxo-7-(2,2,2-trifluoroethyl)--
4,5,6,7-tetrahydro-2H-4.lamda..sup.4-thia-1,2,7-triaza-azulen-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-6,6-dimethyl-7-(2,2,2-trifluoroethy-
l)-6,7-dihydro-2H,5H-4-thia-1,2,7-triaza-azulen-8-one;
2-(2-chlorophenyl)-3-(4-chlorophenyl)-7-(2,2,2-trifluoroethyl)-6,7-dihydr-
o-3H,5H-4-oxa-1,3,7-triaza-azulen-8-one;
2-(2-chlorophenyl)-3-(4-chlorophenyl)-7-(2,2-difluoropropyl)-6,7-dihydro--
3H,5H-4-oxa-1,3,7-triaza-azulen-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-6,6-dimethyl-7-(2,2,2-trifluoroethy-
l)-6,7-dihydro-2H,5H-4-oxa-1,2,7-triaza-azulen-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-7-(2,2,2-trifluoroethyl)-4,5,6,7-te-
trahydro-3H-imidazo[4,5-e][1,4]diazepin-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4-methyl-7-(2,2,2-trifluoroethyl)-4-
,5,6,7-tetrahydro-3H-imidazo[4,5-e][1,4]diazepin-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4,6,6-trimethyl-7-(2,2,2-trifluoroe-
thyl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-e][1,4]diazepin-8-one;
2-(2-chlorophenyl)-3-(4-chlorophenyl)-7-(2,2,2-trifluoroethyl)-6,7-dihydr-
o-3H,5H-4-thia-1,3,7-triaza-azulen-8-one;
2-(2-chlorophenyl)-3-(4-chlorophenyl)-4-oxo-7-(2,2,2-trifluoroethyl)-4,5,-
6,7-tetrahydro-3H-4.lamda..sup.4-thia-1,3,7-triaza-azulen-8-one;
2-(2-chlorophenyl)-3-(4-chlorophenyl)-4,4-dioxo-7-(2,2,2-trifluoroethyl)--
4,5,6,7-tetrahydro-3H-4.lamda..sup.4-thia-1,3,7-triaza-azulen-8-one;
and
2-(2-chlorophenyl)-3-(4-chlorophenyl)-6,6-dimethyl-7-(2,2,2-trifluoroethy-
l)-6,7-dihydro-3H,5H-4-thia-1,3,7-triaza-azulen-8-one; a
pharmaceutically acceptable salt thereof, or a solvate or hydrate
of the compound, or the salt.
[0027] Some of the compounds described herein contain at least one
chiral center; consequently, those skilled in the art will
appreciate that all stereoisomers (e.g., enantiomers and
diastereoisomers) of the compounds illustrated and discussed herein
are within the scope of the present invention. In addition,
tautomeric forms of the compounds are also within the scope of the
present invention.
[0028] Compounds of the present invention have been shown to be
useful cannabinoid receptor ligands (in particular, CB1 receptor
antagonists). Accordingly, another aspect of the present invention
is a pharmaceutical composition that comprises (1) a compound of
the present invention, and (2) a pharmaceutically acceptable
excipient, diluent, or carrier. Preferably, the composition
comprises a therapeutically effective amount of a compound of the
present invention. The composition may also contain at least one
additional pharmaceutical agent (described herein). Preferred
agents include nicotine receptor partial agonists, opioid
antagonists (e.g., naltrexone and nalmefene), dopaminergic agents
(e.g., apomorphine), attention deficit disorder (ADD including
attention deficit hyperactivity disorder (ADHD)) agents (e.g.,
Ritalin.TM., Strattera.TM., Concerta.TM. and Adderall.TM.), and
anti-obesity agents (described herein below).
[0029] In yet another embodiment of the present invention, a method
for treating a disease, condition or disorder modulated by a
cannabinoid receptor (preferably, a CB1 receptor) antagonists in
animals that includes the step of administering to an animal in
need of such treatment a therapeutically effective amount of a
compound of the present invention (or a pharmaceutical composition
thereof).
[0030] Diseases, conditions, and/or disorders modulated by
cannabinoid receptor antagonists include eating disorders (e.g.,
binge eating disorder, anorexia, and bulimia), weight loss or
control (e.g., reduction in calorie or food intake, and/or appetite
suppression), obesity, depression, atypical depression, bipolar
disorders, psychoses, schizophrenia, behavioral addictions,
suppression of reward-related behaviors (e.g., conditioned place
avoidance, such as suppression of cocaine- and morphine-induced
conditioned place preference), substance abuse, addictive
disorders, impulsivity, alcoholism (e.g., alcohol abuse, addiction
and/or dependence including treatment for abstinence, craving
reduction and relapse prevention of alcohol intake), tobacco abuse
(e.g., smoking addiction, cessation and/or dependence including
treatment for craving reduction and relapse prevention of tobacco
smoking), dementia (including memory loss, Alzheimer's disease,
dementia of aging, vascular dementia, mild cognitive impairment,
age-related cognitive decline, and mild neurocognitive disorder),
sexual dysfunction in males (e.g., erectile difficulty), seizure
disorders, epilepsy, inflammation, gastrointestinal disorders
(e.g., dysfunction of gastrointestinal motility or intestinal
propulsion), attention deficit disorder (ADD/ADHD), Parkinson's
disease, and type II diabetes. In a preferred embodiment, the
method is used in the treatment of weight loss, obesity, bulimia,
ADD/ADHD, dementia, alcoholism, and/or tobacco abuse.
[0031] Compounds of the present invention may be administered in
combination with other pharmaceutical agents. Preferred
pharmaceutical agents include nicotine receptor partial agonists,
opioid antagonists (e.g., naltrexone (including naltrexone depot),
antabuse, and nalmefene), dopaminergic agents (e.g., apomorphine),
ADD/ADHD agents (e.g., methylphenidate hydrochloride (e.g.,
Ritalin.TM. and Concerta.TM.), atomoxetine (e.g., Strattera.TM.),
and amphetamines (e.g., Adderall.TM.)) and anti-obesity agents,
such as apo-B/MTP inhibitors, 11.beta.-hydroxy steroid
dehydrogenase-1 (11.beta.-HSD type 1) inhibitors, peptide
YY.sub.3-36 or analogs thereof, MCR-4 agonists, CCK-A agonists,
monoamine reuptake inhibitors, sympathomimetic agents, 3 adrenergic
receptor agonists, dopamine receptor agonists,
melanocyte-stimulating hormone receptor analogs, 5-HT2c receptor
agonists, melanin concentrating hormone receptor antagonists,
leptin, leptin analogs, leptin receptor agonists, galanin receptor
antagonists, lipase inhibitors, bombesin receptor agonists,
neuropeptide-Y receptor antagonists, thyromimetic agents,
dehydroepiandrosterone or analogs thereof, glucocorticoid receptor
antagonists, orexin receptor antagonists, glucagon-like peptide-1
receptor agonists, ciliary neurotrophic factors, human
agouti-related protein antagonists, ghrelin receptor antagonists,
histamine 3 receptor antagonists or inverse agonists, and
neuromedin U receptor agonists, and the like.
[0032] The combination therapy may be administered as (a) a single
pharmaceutical composition which comprises a compound of the
present invention, at least one additional pharmaceutical agent
described above and a pharmaceutically acceptable excipient,
diluent, or carrier; or (b) two separate pharmaceutical
compositions comprising (i) a first composition comprising a
compound of the present invention and a pharmaceutically acceptable
excipient, diluent, or carrier, and (ii) a second composition
comprising at least one additional pharmaceutical agent described
above and a pharmaceutically acceptable excipient, diluent, or
carrier. The pharmaceutical compositions may be administered
simultaneously or sequentially and in any order.
[0033] In yet another aspect of the present invention, a
pharmaceutical kit is provided for use by a consumer to treat
diseases, conditions and/or disorders modulated by cannabinoid
receptor antagonists in an animal. The kit comprises a) a suitable
dosage form comprising a compound of the present invention; and b)
instructions describing a method of using the dosage form to treat
diseases linked to the modulation of the cannabinoid receptor
(preferably, the CB1 receptor).
[0034] In yet another embodiment of the present invention is a
pharmaceutical kit comprising: a) a first dosage form comprising
(i) a compound of the present invention and (ii) a pharmaceutically
acceptable carrier, excipient or diluent; b) a second dosage form
comprising (i) an additional pharmaceutical agent described above,
and (ii) a pharmaceutically acceptable carrier, excipient or
diluent; and c) a container.
Definitions
[0035] As used herein, the term "alkyl" refers to a hydrocarbon
radical of the general formula C.sub.nH.sub.2n+1. The alkane
radical may be straight or branched. For example, the term
"(C.sub.1-C.sub.6)alkyl" refers to a monovalent, straight, or
branched aliphatic group containing 1 to 6 carbon atoms (e.g.,
methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl,
t-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl,
neopentyl, 3,3-dimethylpropyl, hexyl, 2-methylpentyl, and the
like). Similarly, the alkyl portion (i.e., alkyl moiety) of an
alkoxy, acyl (e.g., alkanoyl), alkylamino, dialkylamino, and
alkylthio group have the same definition as above. When indicated
as being "optionally substituted", the alkane radical or alkyl
moiety may be unsubstituted or substituted with one or more
substituents (generally, one to three substituents except in the
case of halogen substituents such as perchloro or perfluoroalkyls)
independently selected from the group of substituents listed below
in the definition for "substituted." "Halo-substituted alkyl"
refers to an alkyl group substituted with one or more halogen atoms
(e.g., "fluoro-substituted alkyl" refers to fluoromethyl,
difluoromethyl, trifluoromethyl, 1-fluoroethyl, 2-fluoroethyl,
1,1-difluoroethyl, 1,2-difluoroethyl, 2,2-difluoroethyl,
1,1,1-trifluoroethyl, 2,2,2-trifluoroethyl, 1,1,2-trifluoroethyl,
1,2,2-trifluoroethyl, 1,2,2,2-tetrafluoroethyl,
1,1,2,2-tetrafluoroethyl, 1,1,1,2-tetrafluoroethyl,
1,1,2,2,2-pentafluoroethyl, 1,1,1,2,2-pentafluoroethyl,
perfluoroethyl, etc.). Preferred halo-substituted alkyls are the
chloro- and fluoro-substituted alkyls, more preferably,
fluoro-substituted alkyls. When substituted, the alkane radicals or
alkyl moieties are preferably fluoro substituents (as described
above), or 1 or 2 substituents independently selected from
(C.sub.1-C.sub.3)alkyl, (C.sub.3-C.sub.6)cycloalkyl,
(C.sub.2-C.sub.3)alkenyl, aryl, heteroaryl, 3- to 6-membered
heterocycle, chloro, cyano, hydroxy, (C.sub.1-C.sub.3)alkoxy,
aryloxy, amino, (C.sub.1-C.sub.6)alkyl amino,
di-(C.sub.1-C.sub.4)alkyl amino, aminocarboxylate (i.e.,
(C.sub.1-C.sub.3)alkyl-O--C(O)--NH--),
hydroxy(C.sub.2-C.sub.3)alkylamino, or keto (oxo), and more
preferably, 1 to 3 fluoro groups, or 1 substituent selected from
(C.sub.1-C.sub.3)alkyl, (C.sub.3-C.sub.6)cycloalkyl, (C.sub.6)aryl,
6-membered-heteroaryl, 3- to 6-membered heterocycle,
(C.sub.1-C.sub.3)alkoxy, (C.sub.1-C.sub.4)alkyl amino or
di-(C.sub.1-C.sub.2)alkyl amino.
[0036] The terms "partially or fully saturated carbocyclic ring"
(also referred to as "partially or fully saturated cycloalkyl")
refers to nonaromatic rings that are either partially or fully
hydrogenated and may exist as a single ring, bicyclic ring or a
spiral ring. Unless specified otherwise, the carbocyclic ring is
generally a 3- to 8-membered ring. For example, partially or fully
saturated carbocyclic rings (or cycloalkyl) include groups such as
cyclopropyl, cyclopropenyl, cyclobutyl, cyclobutenyl, cyclopentyl,
cyclpentenyl, cyclopentadienyl, cyclohexyl, cyclohexenyl,
cyclohexadienyl, norbornyl (bicyclo[2.2.1]heptyl), norbornenyl,
bicyclo[2.2.2]octyl, and the like. When designated as being
"optionally substituted", the partially saturated or fully
saturated cycloalkyl group may be unsubstituted or substituted with
one or more substituents (typically, one to three substituents)
independently selected from the group of substituents listed below
in the definition for "substituted." A substituted carbocyclic ring
also includes groups wherein the carbocyclic ring is fused to a
phenyl ring (e.g., indanyl). The carbocyclic group may be attached
to the chemical entity or moiety by any one of the carbon atoms
within the carbocyclic ring system. When substituted, the
carbocyclic group is preferably substituted with 1 or 2
substituents independently selected from (C.sub.1-C.sub.3)alkyl,
(C.sub.2-C.sub.3)alkenyl, (C.sub.1-C.sub.6)alkylidenyl, aryl,
heteroaryl, 3- to 6-membered heterocycle, chloro, fluoro, cyano,
hydroxy, (C.sub.1-C.sub.3)alkoxy, aryloxy, amino,
(C.sub.1-C.sub.6)alkyl amino, di-(C.sub.1-C.sub.4)alkyl amino,
aminocarboxylate (i.e., (C.sub.1-C.sub.3)alkyl-O--C(O)--NH--),
hydroxy(C.sub.2-C.sub.3)alkylamino, or keto (oxo), and more
preferably 1 or 2 from substituents independently selected from
(C.sub.1-C.sub.2)alkyl, 3- to 6-membered heterocycle, fluoro,
(C.sub.1-C.sub.3)alkoxy, (C.sub.1-C.sub.4)alkyl amino or
di-(C.sub.1-C.sub.2)alkyl amino. Similarly, any cycloalkyl portion
of a group (e.g., cycloalkylalkyl, cycloalkylamino, etc.) has the
same definition as above.
[0037] The term "partially saturated or fully saturated
heterocyclic ring" (also referred to as "partially saturated or
fully saturated heterocycle") refers to nonaromatic rings that are
either partially or fully hydrogenated and may exist as a single
ring, bicyclic ring or a spiral ring. Unless specified otherwise,
the heterocyclic ring is generally a 3- to 6-membered ring
containing 1 to 3 heteroatoms (preferably 1 or 2 heteroatoms)
independently selected from sulfur, oxygen and/or nitrogen.
Partially saturated or fully saturated heterocyclic rings include
groups such as epoxy, aziridinyl, tetrahydrofuranyl,
dihydrofuranyl, dihydropyridinyl, pyrrolidinyl,
N-methylpyrrolidinyl, imidazolidinyl, imidazolinyl, piperidinyl,
piperazinyl, pyrazolidinyl, 2H-pyranyl, 4H-pyranyl, 2H-chromenyl,
oxazinyl, morpholino, thiomorpholino, tetrahydrothienyl,
tetrahydrothienyl 1,1-dioxide, and the like. When indicated as
being "optionally substituted", the partially saturated or fully
saturated heterocycle group may be unsubstiuted or substituted with
one or more substituents (typically, one to three substituents)
independently selected from the group of substituents listed below
in the definition for "substituted." A substituted heterocyclic
ring includes groups wherein the heterocyclic ring is fused to an
aryl or heteroaryl ring (e.g., 2,3-dihydrobenzofuranyl,
2,3-dihydroindolyl, 2,3-dihydrobenzothiophenyl,
2,3-dihydrobenzothiazolyl, etc.). When substituted, the heterocycle
group is preferably substituted with 1 or 2 substituents
independently selected from (C.sub.1-C.sub.3)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, (C.sub.2-C.sub.4)alkenyl, aryl,
heteroaryl, 3- to 6-membered heterocycle, chloro, fluoro, cyano,
hydroxy, (C.sub.1-C.sub.3)alkoxy, aryloxy, amino,
(C.sub.1-C.sub.6)alkyl amino, di-(C.sub.1-C.sub.3)alkyl amino,
aminocarboxylate (i.e., (C.sub.1-C.sub.3)alkyl-O--C(O)--NH--), or
keto (oxo), and more preferably with 1 or 2 substituents
independently selected from (C.sub.1-C.sub.3)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, (C.sub.6)aryl, 6-membered-heteroaryl,
3- to 6-membered heterocycle, or fluoro. The heterocyclic group may
be attached to the chemical entity or moiety by any one of the ring
atoms within the heterocyclic ring system. Similarly, any
heterocycle portion of a group (e.g., heterocycle-substituted
alkyl, heterocycle carbonyl, etc.) has the same definition as
above.
[0038] The term "aryl" or "aromatic carbocyclic ring" refers to
aromatic moieties having a single (e.g., phenyl) or a fused ring
system (e.g., naphthalene, anthracene, phenanthrene, etc.). A
typical aryl group is a 6- to 10-membered aromatic carbocyclic
ring(s). When indicated as being "optionally substituted", the aryl
groups may be unsubstituted or substituted with one or more
substituents (preferably no more than three substituents)
independently selected from the group of substituents listed below
in the definition for "substituted." Substituted aryl groups
include a chain of aromatic moieties (e.g., biphenyl, terphenyl,
phenylnaphthalyl, etc.). When substituted, the aromatic moieties
are preferably substituted with 1 or 2 substituents independently
selected from (C.sub.1-C.sub.4)alkyl, (C.sub.2-C.sub.3)alkenyl,
aryl, heteroaryl, 3- to 6-membered heterocycle, bromo, chloro,
fluoro, iodo, cyano, hydroxy, (C.sub.1-C.sub.4)alkoxy, aryloxy,
amino, (C.sub.1-C.sub.6)alkyl amino, di-(C.sub.1-C.sub.3)alkyl
amino, or aminocarboxylate (i.e.,
(C.sub.1-C.sub.3)alkyl-O--C(O)--NH--), and more preferably, 1 or 2
substituents independently selected from (C.sub.1-C.sub.4)alkyl,
chloro, fluoro, cyano, hydroxy, or (C.sub.1-C.sub.4)alkoxy. The
aryl group may be attached to the chemical entity or moiety by any
one of the carbon atoms within the aromatic ring system. Similarly,
the aryl portion (i.e., aromatic moiety) of an aroyl or aroyloxy
(i.e., (aryl)-C(O)--O--) has the same definition as above.
[0039] The term "heteroaryl" or "heteroaromatic ring" refers to
aromatic moieties containing at least one heteratom (e.g., oxygen,
sulfur, nitrogen or combinations thereof) within a 5- to
10-membered aromatic ring system (e.g., pyrrolyl, pyridyl,
pyrazolyl, indolyl, indazolyl, thienyl, furanyl, benzofuranyl,
oxazolyl, imidazolyl, tetrazolyl, triazinyl, pyrimidyl, pyrazinyl,
thiazolyl, purinyl, benzimidazolyl, quinolinyl, isoquinolinyl,
benzothiophenyl, benzoxazolyl, etc.). The heteroaromatic moiety may
consist of a single or fused ring system. A typical single
heteroaryl ring is a 5- to 6-membered ring containing one to three
heteroatoms independently selected from oxygen, sulfur and nitrogen
and a typical fused heteroaryl ring system is a 9- to 10-membered
ring system containing one to four heteroatoms independently
selected from oxygen, sulfur and nitrogen. When indicated as being
"optionally substituted", the heteroaryl groups may be
unsubstituted or substituted with one or more substituents
(preferably no more than three substituents) independently selected
from the group of substituents listed below in the definition for
"substituted." When substituted, the heteroaromatic moieties are
preferably substituted with 1 or 2 substituents independently
selected from (C.sub.1-C.sub.4)alkyl, (C.sub.2-C.sub.3)alkenyl,
aryl, heteroaryl, 3- to 6-membered heterocycle, bromo, chloro,
fluoro, iodo, cyano, hydroxy, (C.sub.1-C.sub.4)alkoxy, aryloxy,
amino, (C.sub.1-C.sub.6)alkyl amino, di-(C.sub.1-C.sub.3)alkyl
amino, or aminocarboxylate (i.e.,
(C.sub.1-C.sub.3)alkyl-O--C(O)--NH--), and more preferably, 1 or 2
substituents independently selected from (C.sub.1-C.sub.4)alkyl,
chloro, fluoro, cyano, hydroxy, (C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkyl amino or di-(C.sub.1-C.sub.2)alkyl amino.
The heteroaryl group may be attached to the chemical entity or
moiety by any one of the atoms within the aromatic ring system
(e.g., imidazol-1-yl, imidazol-2-yl, imidazol-4-yl, imidazol-5-yl,
pyrid-2-yl, pyrid-3-yl, pyrid-4-yl, pyrid-5-yl, or pyrid-6-yl).
Similarly, the heteroaryl portion (i.e., heteroaromatic moiety) of
a heteroaroyl or heteroaroyloxy (i.e., (heteroaryl)-C(O)--O--) has
the same definition as above.
[0040] The term "acyl" refers to hydrogen, alkyl, partially
saturated or fully saturated cycloalkyl, partially saturated or
fully saturated heterocycle, aryl, and heteroaryl substituted
carbonyl groups. For example, acyl includes groups such as
(C.sub.1-C.sub.6)alkanoyl (e.g., formyl, acetyl, propionyl,
butyryl, valeryl, caproyl, t-butylacetyl, etc.),
(C.sub.3-C.sub.6)cycloalkylcarbonyl (e.g., cyclopropylcarbonyl,
cyclobutylcarbonyl, cyclopentylcarbonyl, cyclohexylcarbonyl, etc.),
heterocyclic carbonyl (e.g., pyrrolidinylcarbonyl,
pyrrolid-2-one-5-carbonyl, piperidinylcarbonyl,
piperazinylcarbonyl, tetrahydrofuranylcarbonyl, etc.), aroyl (e.g.,
benzoyl) and heteroaroyl (e.g., thiophenyl-2-carbonyl,
thiophenyl-3-carbonyl, furanyl-2-carbonyl, furanyl-3-carbonyl,
1H-pyrroyl-2-carbonyl, 1H-pyrroyl-3-carbonyl,
benzo[b]thiophenyl-2-carbonyl, etc.). In addition, the alkyl,
cycloalkyl, heterocycle, aryl and heteroaryl portion of the acyl
group may be any one of the groups described in the respective
definitions above. When indicated as being "optionally
substituted", the acyl group may be unsubstituted or optionally
substituted with one or more substituents (typically, one to three
substituents) independently selected from the group of substituents
listed below in the definition for "substituted" or the alkyl,
cycloalkyl, heterocycle, aryl and heteroaryl portion of the acyl
group may be substituted as described above in the preferred and
more preferred list of substituents, respectively.
[0041] The term "substituted" specifically envisions and allows for
one or more substitutions that are common in the art. However, it
is generally understood by those skilled in the art that the
substituents should be selected so as to not adversely affect the
pharmacological characteristics of the compound or adversely
interfere with the use of the medicament. Suitable substituents for
any of the groups defined above include (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.7)cycloalkyl, (C.sub.2-C.sub.6)alkenyl,
(C.sub.1-C.sub.6)alkylidenyl, aryl, heteroaryl, 3- to 6-membered
heterocycle, halo (e.g., chloro, bromo, iodo and fluoro), cyano,
hydroxy, (C.sub.1-C.sub.6)alkoxy, aryloxy, sulfhydryl (mercapto),
(C.sub.1-C.sub.6)alkylthio, arylthio, amino, mono- or
di-(C.sub.1-C.sub.6)alkyl amino, quaternary ammonium salts,
amino(C.sub.1-C.sub.6)alkoxy, aminocarboxylate (i.e.,
(C.sub.1-C.sub.6)alkyl-O--C(O)--NH--),
hydroxy(C.sub.2-C.sub.6)alkylamino,
amino(C.sub.1-C.sub.6)alkylthio, cyanoamino, nitro,
(C.sub.1-C.sub.6)carbamyl, keto (oxo), acyl,
(C.sub.1-C.sub.6)alkyl-CO.sub.2--, glycolyl, glycyl, hydrazino,
guanyl, sulfamyl, sulfonyl, sulfinyl,
thio(C.sub.1-C.sub.6)alkyl-C(O)--,
thio(C.sub.1-C.sub.6)alkyl-CO.sub.2--, and combinations thereof. In
the case of substituted combinations, such as "substituted
aryl(C.sub.1-C.sub.6)alkyl", either the aryl or the alkyl group may
be substituted, or both the aryl and the alkyl groups may be
substituted with one or more substituents (typically, one to three
substituents except in the case of perhalo substitutions). An aryl
or heteroaryl substituted carbocyclic or heterocyclic group may be
a fused ring (e.g., indanyl, dihydrobenzofuranyl, dihydroindolyl,
etc.).
[0042] The term "solvate" refers to a molecular complex of a
compound represented by Formula (I) or (II) (including prodrugs and
pharmaceutically acceptable salts thereof) with one or more solvent
molecules. Such solvent molecules are those commonly used in the
pharmaceutical art, which are known to be innocuous to the
recipient, e.g., water, ethanol, and the like. The term "hydrate"
refers to the complex where the solvent molecule is water.
[0043] The term "protecting group" or "Pg" refers to a substituent
that is commonly employed to block or protect a particular
functionality while reacting other functional groups on the
compound. For example, an "amino-protecting group" is a substituent
attached to an amino group that blocks or protects the amino
functionality in the compound. Suitable amino-protecting groups
include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC),
benzyloxycarbonyl (CBz) and 9-fluorenylmethylenoxycarbonyl (Fmoc).
Similarly, a "hydroxy-protecting group" refers to a substituent of
a hydroxy group that blocks or protects the hydroxy functionality.
Suitable protecting groups include acetyl and silyl. A
"carboxy-protecting group" refers to a substituent of the carboxy
group that blocks or protects the carboxy functionality. Common
carboxy-protecting groups include --CH.sub.2CH.sub.2SO.sub.2Ph,
cyanoethyl, 2-(trimethylsilyl)ethyl,
2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl,
2-(p-nitrophenylsulfenyl)ethyl, 2-(diphenylphosphino)-ethyl,
nitroethyl and the like. For a general description of protecting
groups and their use, see T. W. Greene, Protective Groups in
Organic Synthesis, John Wiley & Sons, New York, 1991.
[0044] The phrase "therapeutically effective amount" means an
amount of a compound of the present invention that (i) treats or
prevents the particular disease, condition, or disorder, (ii)
attenuates, ameliorates, or eliminates one or more symptoms of the
particular disease, condition, or disorder, or (iii) prevents or
delays the onset of one or more symptoms of the particular disease,
condition, or disorder described herein.
[0045] The term "animal" refers to humans (male or female),
companion animals (e.g., dogs, cats and horses), food-source
animals, zoo animals, marine animals, birds and other similar
animal species. "Edible animals" refers to food-source animals such
as cows, pigs, sheep and poultry.
[0046] The phrase "pharmaceutically acceptable" indicates that the
substance or composition must be compatible chemically and/or
toxicologically, with the other ingredients comprising a
formulation, and/or the mammal being treated therewith.
[0047] The terms "treating", "treat", or "treatment" embrace both
preventative, i.e., prophylactic, and palliative treatment.
[0048] The terms "modulated by a cannabinoid receptor" or
"modulation of a cannabinoid receptor" refers to the activation or
deactivation of a cannabinoid receptor. For example, a ligand may
act as an agonist, partial agonist, inverse agonist, antagonist, or
partial antagonist.
[0049] The term "antagonist" includes both full antagonists and
partial antagonists, as well as inverse agonists.
[0050] The term "CB-1 receptor" refers to the G-protein coupled
type 1 cannabinoid receptor.
[0051] The term "compounds of the present invention" (unless
specifically identified otherwise) refer to compounds of Formulae
(I), (II) (I-A), (I-B), (I-C), (I-D), (I-E), (I-F), (I-G), (I-H),
(I-I), (I-J), (I-K), (I-L), (I-M), (I-N), (I-O), (I-P) and (I-Q),
prodrugs thereof, pharmaceutically acceptable salts of the
compounds, and/or prodrugs, and hydrates or solvates of the
compounds, salts, and/or prodrugs, as well as, all stereoisomers
(including diastereoisomers and enantiomers), tautomers and
isotopically labeled compounds. All amorphous and crystalline forms
of the compounds are included as well.
[0052] As used herein, structures drawn with circles within a ring
designate aromaticity. For example, the following chemical moiety
designates a pyrazole ring when A is nitrogen and B is carbon; and
the chemical moiety designates an imidazole when A is carbon and B
is nitrogen. ##STR4##
DETAILED DESCRIPTION
[0053] Compounds of the present invention may be synthesized by
synthetic routes that include processes analogous to those
well-known in the chemical arts, particularly in light of the
description contained herein. The starting materials are generally
available from commercial sources such as Aldrich Chemicals
(Milwaukee, Wis.) or are readily prepared using methods well known
to those skilled in the art (e.g., prepared by methods generally
described in Louis F. Fieser and Mary Fieser, Reagents for Organic
Synthesis, v. 1-19, Wiley, New York (1967-1999 ed.), or Beilsteins
Handbuch der organischen Chemie, 4, Aufl. ed. Springer-Verlag,
Berlin, including supplements (also available via the Beilstein
online database)).
[0054] For illustrative purposes, the reaction schemes depicted
below provide potential routes for synthesizing the compounds of
the present invention as well as key intermediates. For a more
detailed description of the individual reaction steps, see the
Examples section below. Those skilled in the art will appreciate
that other synthetic routes may be used to synthesize the inventive
compounds. Although specific starting materials and reagents are
depicted in the schemes and discussed below, other starting
materials and reagents can be easily substituted to provide a
variety of derivatives and/or reaction conditions. In addition,
many of the compounds prepared by the methods described below can
be further modified in light of this disclosure using conventional
chemistry well known to those skilled in the art.
[0055] In the preparation of compounds of the present invention,
protection of remote functionality (e.g., primary or secondary
amine) of intermediates may be necessary. The need for such
protection will vary depending on the nature of the remote
functionality and the conditions of the preparation methods.
Suitable amino-protecting groups (NH-Pg) include acetyl,
trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBz)
and 9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such
protection is readily determined by one skilled in the art. For a
general description of protecting groups and their use, see T. W.
Greene, Protective Groups in Organic Synthesis, John Wiley &
Sons, New York, 1991.
[0056] Scheme I outlines the general procedures one could use to
provide compounds of the present invention where A is nitrogen, B
is carbon and X is O (i.e., compound of Formula (I-A)).
##STR5##
[0057] The starting pyrazolo ester may be prepared by procedures
described in U.S. Pat. No. 5,624,941 and is incorporated herein by
reference. The bromo intermediate (1a) may be prepared using
conventional bromination procedures well-known to those skilled in
the art. For example, the pyrazolo ester may be treated with
bromine in a protic solvent (e.g., acetic acid) at a temperature
from about 10.degree. C. to about -10.degree. C. A vinyl group may
then be introduced by treating the bromo intermediate (1a) with
tributylvinyltin and tetrakistriphenylphosphine palladium in a
polar solvent (e.g., dimethylformamide (DMF)) at an elevated
temperature. The vinyl group may then be oxidatively cleaved to the
corresponding aldehyde. For example, the vinyl intermediate (1b)
may be treated with osmium tetroxide in the presence of
N-methylmorpholine-N-oxide in an aqueous solvent (e.g., dioxane and
water) at about room temperature followed by treatment with sodium
periodate. The aldehyde group may then be converted to a hydroxy
group by treating aldehyde intermediate (1c) with a percarboxylic
acid (e.g., m-chloroperbenzoic acid) in an aprotic solvent (e.g.,
dichloromethane) followed by treatment with a strong base (e.g.,
triethylamine) in a protic solvent (e.g., methanol). An appropriate
allyl group may be condensed with the hydroxy group to form the
desired allyl ether intermediate (1e) using conventional means. For
example, hydroxy intermediate (1d) may be treated with a strong
base (e.g., sodium hydride) followed by the addition of the desired
allyl bromide in a polar solvent (e.g., dimethylsulfoxide (DMSO)).
The pendant olefin group may be oxidatively cleaved to its
corresponding aldehyde using procedures analogous to those
described above for the conversion of vinyl intermediate (1 b) to
its corresponding aldehyde intermediate (1c). The desired amino
group (--NHR.sup.4) may be introduced by treating aldehyde
intermediate (1e) with the desired amine (R.sup.4NH.sub.2) in the
presence of sodium triacetoxyborohydride (NaBH(OAc).sub.3) in a
protic solvent (e.g., acetic acid and 1,2-dichloroethane). The
carboxy-protecting group may be removed by treating the ester with
a strong base (e.g., an alkali metal hydroxide, such as potassium
hydroxide) in a protic solvent (e.g., ethanol). The amino
intermediate (1h) may then be cyclized to the final product (1-A)
by treatment with 1-propanephosphoric acid cyclic anhydride in the
presence of a base (e.g., triethylamine) in an aprotic solvent
(1,2-dichloroethane).
[0058] Scheme II below illustrates an alternative synthesis of
intermediate (1d). ##STR6##
[0059] The keto ester intermediate (2a) can be prepared by
condensing the desired acid chloride with
2,2-dimethyl-[1,3]dioxane-4,6-dione in the presence of a base
(e.g., pyridine) in an aprotic solvent (e.g., methylene chloride)
followed by heating at an elevated temperature in a protic solvent
(e.g., ethanol). The hydrazono intermediate (2b) can then be
prepared by treating the keto ester (2a) with the desired amine in
the presence of sodium nitrate in an acidic medium (e.g., aqueous
acetic acid). The bromo group may then be introduced using standard
bromination procedures well-known to those skilled in the art. For
example, intermediate (2b) can be treated with copper (II) bromide
in an aprotic solvent (e.g., ethyl acetate and chloroform) at an
elevated temperature. Cyclization of the bromo intermediate (2c)
may then be accomplished by heating in a polar solvent (e.g.,
methanol) in the presence of sodium acetate (or triethylamine).
[0060] Scheme III outlines an alternative procedure for
synthesizing compounds of the present invention where A is
nitrogen, B is carbon and X is O (i.e., compound of Formula (I-B))
starting with intermediate (1d). ##STR7##
[0061] The amide intermediate (3a) may be prepared from the
carboxylic ester (1d) by condensing the desired hydroxy amino
compound with intermediate (1e) at an elevated temperature in an
aprotic solvent (e.g., toluene). The compound of Formula (I-B) may
then be produced using standard ether-forming reactions well-known
to those skilled in the art. For example, the ether linkage may be
formed using the Mitsunobu reaction conditions
(1,1'-(azodicarbonyl)dipiperidine (ADDP) in the presence of
triphenylphosphine). See, Mitsunobu, O., Synthesis, 1 (1981).
[0062] Scheme IV outlines the general procedures one could use to
provide compounds of the present invention where A is nitrogen, B
is carbon, and X is --C(R.sup.2b)(R.sup.2c)--, where R.sup.2b and
R.sup.2c are as defined above. ##STR8##
[0063] The allyl group of intermediate (4a) may be introduced via a
palladium (Pd(0)) catalyzed coupling of the bromo intermediate (1a)
with the desired organostannane using procedures analogous to those
described by Martorell, G., et al. in "Palladium catalyzed
cross-coupling of phenol triflates with organostannanes. A
versatile approach for the synthesis of substituted resorcinol
dimethyl ethers," Tetrahedron Lett, 31(16), 2357-2360 (1990). For
example, intermediate (1a) may be treated with the desired
organostannane (e.g., allyl-SnBu.sub.3) in the presence of a
palladium catalyst (e.g., Pd(0)/phosphine (e.g.,
triphenylphosphine, 1,1'-bisdiphenylphosphinoferrocene (dppf),
1,3-bisdiphenyl-phosphinoopropane (dppp) or
1,2-bisdiphenylphosphinoethane (dppe))/lithium chloride) in
refluxing DMF. The carboxy-protecting group may be removed using
conventional methods well-known to those skilled in the art, such
as treatment with a strong alkali base (e.g., potassium hydroxide)
in a protic solvent (e.g., ethanol). The carboxy group may then be
condensed with the desired amine (R.sup.4NH.sub.2) to produce the
amide intermediate (4c). For example, carboxylic acid intermediate
(4b) may be treated with R.sup.4NH.sub.2 in the presence of
1-propanephosphoric acid cyclic anhydride and a base (e.g.,
triethylamine) in an aprotic solvent (e.g., 1,2-dichloroethane).
The vinyl group of the amide intermediate (4c) may be hydrated
using conventional means well-known to those skilled in the art.
For example, amide intermediate (4c) may be treated with
9-borabicyclo[3.3.1]nonane (9-BBN) in an aprotic solvent (e.g.,
tetrahydrofuran (THF)) followed by the addition of hydrogen
peroxide and aqueous sodium hydroxide. The resultant hydroxy
intermediate (4d) may then be sulfonated by reacting the hydroxy
group with an alkyl sulfonyl chloride (e.g., R'SO.sub.2Cl) in the
presence of a base (e.g., triethylamine) and an aprotic solvent
(methylene chloride). The sulfonate intermediate (4e) may be
cyclized to the final product (I-C) by treating with a strong base
(e.g., sodium hydride) in an aprotic solvent (e.g., THF).
[0064] Scheme V outlines the general procedures one could use to
provide compounds of the present invention where A is nitrogen, B
is carbon, and X is --N(R.sup.2a)--, where R.sup.2a is as defined
above. ##STR9##
[0065] Lithium salt (5b) can be prepared by treatment of methyl
ketone (5a) with lithium hexamethyldisilazide at a temperature of
about -78.degree. C. in an aprotic solvent such as THF, followed by
condensation with diethyl oxalate, as described in WO 00/46209. The
isolated lithium salt (5b) is then dissolved in an acid such as
acetic acid and nitrosated by dropwise addition of aqueous sodium
nitrite at a temperature of about 0-10.degree. C. (Tetrahedron, 3,
209 (1958); Bull. Chem. Soc. Jpn. 52, 208 (1979)). A substituted
hydrazine may then be added directly to the reaction mixture to
afford intermediate (5c). Cyclization of (5c) is accomplished by
heating intermediate (5c) and a catalytic amount of an acid such as
concentrated sulfuric acid in a solvent such isopropanol at a
temperature of about 60.degree. C. to provide nitrosopyrazole (5d).
The nitroso group of intermediate (5d) can be reduced by treatment
of (5d) with sodium dithionite in a mixture of solvents such as
ethyl acetate and water, affording aminopyrazole (5e), which may be
reductively alkylated with an appropriately protected amino
aldehyde (5f) (such as tert-butyl N-(2-oxoethyl)carbamate when
R.sup.4.dbd.H) and a reagent like sodium triacetoxyborohydride or
sodium cyanoborohydride to give intermediate (5g) (see, e.g., EP
1329160). Alternatively, amine (5e) can be coupled with acid (5h)
under standard conditions to give amide (5i), which can then be
reduced (e.g., BH.sub.3) to give amine (5g). The carboxy-protecting
group in (5g) can be hydrolyzed with a strong base (e.g., an alkali
metal hydroxide such as potassium hydroxide) in a polar, protic
solvent (e.g., ethanol) to give intermediate (5j). Removal of the
amino protecting group using standard methods (e.g.,
trifluoroacetic acid or aqueous HCl in ethanol for removal of Boc
group, hydrogenolysis for removal of Cbz group) can provide the
amino acid derivative (5k) which can cyclize in the presence of a
coupling reagent (e.g., EDC or
O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (HATU)) to form the lactam (5m). Alternatively,
deprotection of the amino functionality in (5g) using standard
conditions, followed by treatment with a base (e.g., sodium
ethoxide) in an alcoholic solvent (e.g., ethanol) or with a strong
acid such as polyphosphoric acid (PPA) can yield lactam (5m).
Compounds of formula (I-D) can be prepared from (5m) by treatment
with a suitable alkylating agent (i.e., R.sup.4--X where X is a
leaving group) in the presence of a strong base (e.g., sodium
hydride) in a polar solvent (e.g., DMF, THF). Compounds of formula
(I-E) can be prepared from (I-D) by deprotonation with a base such
as sodium hydride or sodium hexamethyldisilazide in a solvent such
as DMF, followed by alkylation with R.sup.2a--X. Treatment of (I-D)
with an appropriately substituted acyl chloride or acyl anhydride
in the presence of a tertiary base such as pyridine, triethylamine,
or diisopropylethylamine in a non-polar solvent such as
CH.sub.2Cl.sub.2 or benzene can provide compounds of Formula (I-E)
where R.sup.2a is (C.sub.1-C.sub.4)alkylcarbonyl. In certain
instances, it maybe necessary to first protect the N-4 amino group
in intermediate (5m) with a trifluoroacetyl moiety which can be
removed in a subsequent step following alkylation with R.sup.4--X.
The R.sup.2, group in (I-E) can also be introduced earlier in the
sequence by treating intermediate (5g) with an appropriately
substituted acylating agent in the presence of base (e.g., DMAP,
pyridine) in a non-polar solvent (e.g., CH.sub.2Cl.sub.2). If
necessary, the amide moiety in intermediate (5g) can be selectively
reduced in the presence of the ester functionality using BH.sub.3
in a polar solvent (e.g., THF). Deprotection of (5g) and
cyclization as previously described can provide compounds of
formula (I-E).
[0066] Representative examples of compounds of Formula (I-D) that
may be prepared by the procedures described above in Scheme IV
include:
3-(4-chlorophenyl)-2-(2-chlorophenyl)-7-(2,2,2-trifluoroethyl)-4,5,6,7-te-
trahydro-2H-pyrazolo[4,3-e][1,4]diazepin-8-one; and
3-(4-chlorophenyl)-2-(2-chlorophenyl)-6,6-dimethyl-7-(2,2,2-trifluoroethy-
l)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-e][1,4]diazepin-8-one.
[0067] A representative example of a compound of Formula (I-E) that
may be prepared by the procedures described above in Scheme IV
includes
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4,6,6-trimethyl-7-(2,2,2-trifluoroe-
thyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-e][1,4]diazepin-8-one.
[0068] Scheme VI outlines a variation in the procedure described in
Scheme V where the R.sup.4 substituent is already present in the
starting aldehyde (6a) or acid (6c). ##STR10##
[0069] The aminopyrazole intermediate (5e) can be treated with an
aldehyde intermediate (6a) in the presence of an acid catalyst
(e.g. acetic acid) and reducing agent (e.g. NaBH.sub.3CN,
NaBH(OAc).sub.3) in a non-polar solvent (e.g. 1,2-dichloroethane)
to give intermediate (6b). Alternatively, amine (5e) can be
acylated with acid (6c) under standard conditions to give amide
(6d), which can then be reduced (e.g., BH.sub.3) to give amine
(6b). Removal of the amino protecting group under standard
conditions can yield intermediate (6e) that can be converted into
lactam (I-F) as previously described. Representative examples of
compounds of Formula (I-F) that may be prepared by the procedures
described above in Scheme VI include
2-(2-chlorophenyl)-1-(4-chlorophenyl)-9-methyl-5,6,7,7a,8,9-hexahydro-2H--
2,3,4a,9-tetraazacyclopenta[f]azulen-4-one; and
2-(2-chlorophenyl)-1-(4-chlorophenyl)-2,5,6,7,8,8a,9,10-octahydro-2,3,4a,-
10-tetraaza-benzo[f]azulen-4-one.
[0070] Scheme VII outlines an alternative procedure to prepare
compounds of the present invention where A is nitrogen, B is
carbon, R.sup.3a is hydrogen and R.sup.3b is hydrogen, and X is
--N(R.sup.2a)--, where R.sup.2a is as defined above. ##STR11##
[0071] Intermediate (7a) may be prepared from amine (5e) using
conventional reductive amination procedures well known to those
skilled in the art. Intermediate (7a) may also be prepared by
acylation of (5e), followed by selective amide reduction as
described earlier. The amine may then be homologated to bromide
(7b), such as by alkylation with 1,2-dibromoethane in the presence
of a base like potassium carbonate or sodium carbonate in a
suitable solvent (e.g., acetonitrile, THF, or DMF) at a temperature
from around 0-100.degree. C. using procedures such as those
described in Heteroatom Chemistry, 13, 63-71 (2002). Subsequent
treatment with a primary amine will give an intermediate secondary
amine (see U.S. Pat. No. 6,207,663) which may spontaneously cyclize
to the lactam (I-G). Alternatively, the ester can be hydrolyzed to
the acid and then coupled with the amine to form the lactam (I-G)
using conditions well-known to those skilled in the art. A
representative example of compound of Formula (I-G) that may be
prepared by the procedures described above in Scheme VII includes
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4-methyl-7-(2,2,2-trifluoroethyl)-4-
,5,6,7-tetrahydro-2H-pyrazolo[4,3-e][1,4]diazepin-8-one.
[0072] Compounds of the present invention where A is carbon, B is
nitrogen, R.sup.3a and R.sup.3b are hydrogen, and X is O can be
prepared as shown in Scheme VIII. ##STR12##
[0073] Intermediate (8a) can be reacted with dialkyl aminomalonate
(8b) to provide the 5-hydroxy-1H-imidazole-4-carboxylic acid alkyl
ester derivative (8c) using procedures analogous to those described
in J. Het. Chem. 19, 193-200 (1982). Treatment of (8c) with allyl
bromide in a polar solvent (e.g., THF) using a mild base (e.g.,
potassium carbonate) can provide the O-alkylated intermediate (8d).
The olefin group in (8d) can be di-hydroxylated using osmium
tetroxide in the presence of N-methylmorpholine-N-oxide in an
aqueous solvent (e.g. dioxane and H.sub.2O) and the diol
intermediate can be oxidatively cleaved using sodium periodate to
give intermediate (8e). Treatment of the aldehyde (8e) with an
appropriately substituted amine (R.sup.4NH.sub.2) in the presence
of an acid catalyst (e.g. HOAc) and a reducing agent (e.g.,
NaBH.sub.3CN, NaBH(OAc).sub.3) can yield intermediate (8f).
Hydrolysis of the carboxy-protecting group in (8f) can provide
intermediate (8g) which can be cyclized in the presence of EDC or
HATU as previously described to provide compound (I-H).
Intermediate (8f) can be converted directly into (I-H) under acidic
(PPA) or basic (NaOEt/EtOH) conditions as described earlier.
[0074] In an alternative synthesis of compounds of formula (I-H)
wherein R.sup.3a and R.sup.3b may be a substituent other than
hydrogen, intermediate (8c) can be reacted with a substituted or
unsubstituted .beta.-haloethyleneamine derivative where the amino
group is suitably protected and R.sup.4a is either hydrogen or
R.sup.4. The product from the alkylation step, intermediate (8i),
can be deprotected under standard conditions and cyclized in the
presence of acid (e.g., PPA) or base (e.g., NaOEt/EtOH) to yield
(I-H) when R.sup.4a is R.sup.4. When R.sup.4a is hydrogen, the
amide may be alkylated under standard conditions as described above
to provide (I-H).
[0075] Scheme IX describes an alternative method for preparing
intermediate (8c). ##STR13##
[0076] Intermediate (9a) can be prepared by treating the
appropriate amine having the desired R.sup.1 group with
trimethylaluminum under inert atmospheric conditions followed by
condensation with the appropriate cyanide having the desired
R.sup.0 group. Suitable amines include substituted phenyl amines
(e.g., 4-chlorophenyl amine, 4-fluorophenyl amine, 4-bromophenyl
amine, 4-iodophenyl amine, 4-cyanophenyl amine, and the like)
pyridin-2-yl amine, pyridin-3-yl amine, pyridin-4-yl amine,
substituted pyridinyl amines (e.g., 2-dimethylaminopyridin-5-yl
amine, 2-methoxypyridin-5-yl amine, 5-chloropyridin-2-yl amine,
5-methylpyridin-2-yl, 5-methoxypyridin-2-yl amine,
3-chloropyridin-4-yl; amine, 2-N-morpholinylpyridin-5-yl, and the
like), and other commercially available or easily synthesized
substituted or unsubstituted aryl and heteroaryl amines. Suitable
cyano compounds include substituted benzonitriles (e.g.,
2-chlorobenzonitrile, 2-fluorobenzonitrile, 2-methoxybenzonitrile,
2-methylbenzonitrile, 2,4-dichlorobenzonitrile,
2,4-difluorobenzonitrile, 2-chloro-4-fluorobenzonitrile,
2-chloro-4-methylbenzonitrile, 2,4-dimethoxybenzonitrile,
2-methyl-4-chlorobenzonitrile, and the like), cyano-substituted
pyridines (e.g., 4-cyano-3-chloropyridine) and other commercially
available or easily synthesized substituted or unsubstituted aryl
or heteroaryl nitriles.
[0077] Intermediate (9a) can then be condensed with a
3-bromo-2-oxo-propionic acid ester (wherein R is an alkyl group
like methyl, ethyl, propyl, benzyl, etc.) to produce the cyclized
4-hydroxy-4,5-dihydro-1H-imidazole ester (9b) using procedures
analogous to those described by Khanna, I. K., et al., in J. Med.
Chem., 40, 1634 (1997). For example, the amidine intermediate (8a)
is refluxed in a polar solvent (e.g., isopropanol) in the presence
of a mild base (e.g., sodium bicarbonate). Generally, the reaction
(i.e., cyclization followed by dehydration) proceeds directly to
the desired imidazole ester intermediate (9c). In some instances,
it may be necessary to dehydrate the carbinol condensation product
(9b) with an acid catalyst (e.g., toluene sulfonic acid in
refluxing toluene) to provide the desired imidazole ester (9c).
[0078] The imidazole ester (9c) can be prepared from the
4-hydroxy-4,5-dihydro intermediate (9b) using standard dehydration
procedures well-known to those skilled in the art. For example,
intermediate (9b) may be treated with p-toluenesulfonic acid
monohydrate in refluxing toluene. Alternatively, intermediate (9b)
may be treated with methanesulfonyl chloride in the presence of a
base (e.g., triethylamine).
[0079] Intermediate (9d, where L.sup.1 is a halogen) can be
synthesized from imidazole ester (9c) using a halogenating agent
such as bromine, N-bromosuccinimide, iodine, or N-iodosuccinimide
in a suitable protic solvent such as glacial acetic acid or
trifluoroacetic acid or an aprotic solvent such as acetonitrile,
ether or THF, at reaction temperatures ranging from 35.degree. C.
to 100.degree. C. Transmetalation of (9d) using an alkyl lithium
base, preferentially n-BuLi or tert.-butyl lithium, or an alkyl
Grignard reagent such MeMgBr or EtMgBr, in a polar, aprotic solvent
such as diethyl ether, dioxane or THF, at reaction temperatures
ranging from -100.degree. C. to -78.degree. C., followed by
treatment with a formyl equivalent such as DMF, formylpiperidine,
or ethyl formate provides the aldehyde derivative (9e).
Alternatively, (8e) may be prepared directly from (9c) by: (1)
treatment with POCl.sub.3 or POBr.sub.3 in a solvent such as DMF at
reaction temperatures ranging from 35.degree. C. to 100.degree. C.,
followed by hydrolysis; or (2) preparing in situ several
equivalents of a Vilsmeier reagent (POCl.sub.3 or POBr.sub.3 in
DMF) in an aprotic solvent such as CH.sub.2Cl.sub.2 or
dichloroethane, followed by hydrolysis. The aldehyde intermediate
(9e) can be converted into the hydroxy intermediate (8c) by the
methods described in Scheme I for the conversion of intermediate
(1c) to intermediate (1d).
[0080] The aldehyde intermediate (9e) may also be synthesized by
the general route shown in Scheme X below. ##STR14##
[0081] Intermediate (9a) can be condensed with
3-bromo-4-hydroxy-2-oxo-butyric acid alkyl ester derivative (10a)
with a suitable protecting group on the 4-hydroxy substituent to
produce the cyclized intermediate (10b) which can be dehydrated as
previously described to give the imidazoyl intermediate (10c). The
protecting group (Pg) on the 5-hydroxymethyl group in intermediate
(10c) can be subsequently removed by standard procedures well known
to those skilled in the art. Intermediate (10d) may then be
transformed into (9e) using oxidation procedures analogous to those
described in Tett. Lett., 35, 9391-4 (1994) or J. Het. Chem., 39,
841-844 (2002). For example, the 5-hydroxymethylimidazolyl
derivative (9d) can be treated with oxalyl chloride, DMSO and a
tertiary amine base such as triethylamine or diisopropylethylamine
in a halogenated solvent such as CH.sub.2Cl.sub.2 or CHCl.sub.3.
Alternatively, the intermediate (10d) can be oxidized using
MnO.sub.2 in a polar or non-polar solvent such as MeOH, acetone,
dioxane, ether, CH.sub.2Cl.sub.2, or CHCl.sub.3.
[0082] Representative examples of compounds of Formula (I-H) that
may be prepared by the procedures described above in Schemes VIII,
IX or X include:
2-(2-chlorophenyl)-3-(4-chlorophenyl)-7-(2,2,2-trifluoroethyl)-6-
,7-dihydro-3H,5H-4-oxa-1,3,7-triaza-azulen-8-one;
2-(2-chlorophenyl)-3-(4-chlorophenyl)-7-(2,2-difluoropropyl)-6,7-dihydro--
3H,5H-4-oxa-1,3,7-triaza-azulen-8-one; and
3-(4-chlorophenyl)-2-(2-chlorophenyl)-6,6-dimethyl-7-(2,2,2-trifluoroethy-
l)-6,7-dihydro-2H,5H-4-oxa-1,2,7-triaza-azulen-8-one.
[0083] Compounds of the present invention where A is carbon, B is
nitrogen and X is N can be prepared as shown in Scheme XI.
##STR15##
[0084] The intermediate (11a), prepared as shown in Scheme IX for
compound (9d, L=Br), can be treated with an ammonia equivalent such
as lithium bis(trimethylsilyl)amide in the presence of a catalytic
amount of Pd(dba).sub.2 and a phosphine ligand such as
P(t-Bu).sub.3 in a non-polar solvent (e.g., toluene) at
temperatures ranging from 23.degree. C. to reflux to give
intermediate (11 b). Examples of related procedures are described
by Lee et al. in Organic Letters, 3, 2729-273 (2001). Treatment of
(11b) with an appropriately protected amino aldehyde derivative
(11c) (such as tert.-butyl N-(2-oxoethyl)carbamate when
R.sup.4a.dbd.H), wherein R.sup.4a is either hydrogen or R.sup.4,
and a reducing reagent (e.g., sodium triacetoxyborohydride or
sodium cyanoborohydride) can provide intermediate (11e) (see, e.g.,
EP 1329160). Alternatively, amine (11b) can be acylated with acid
(11d) under standard conditions to give an amide, which can then be
reduced (e.g., BH.sub.3) to give amine (11e) as described above.
Hydrolysis of the carboxy-protecting group in (11e) with aqueous
K.sub.2CO.sub.3 in an alcoholic solvent or with an alkali base such
as KOH in a polar solvent (e.g., ethanol) can provide intermediate
(11f). Deprotection of the amino group in (11f) using standard
methods can provide intermediate (11g), which can undergo
cyclization as previously described to form lactam (I-I,
R.sup.4a.dbd.H) or compound (I-J, R.sup.4a.dbd.R.sup.4).
Alternatively, the amino protecting group in (11e) can be removed
and the product treated with a base (e.g., sodium methoxide) in an
alcoholic solvent (e.g., methanol) or an acid (e.g., PPA) to form
the lactam (I-I, R.sup.4a.dbd.H) or compound (I-J,
R.sup.4a.dbd.R.sup.4) as described previously. Compounds of formula
(I-J) and (I-K) can be prepared from (I-I) as described for
pyrazolyl analogs (I-D) and (I-E) in Schemes V-VII.
[0085] Compounds of formula (I-J) can also be prepared from
compound (I-I) via the route shown in Scheme XII. ##STR16##
[0086] Reaction of intermediate (11a) with ethylenediamine or an
appropriately substituted ethylenediamine derivative and cupric
oxide in a solvent such as pyridine in the presence of a base such
as potassium carbonate can provide intermediate (12a) which can
undergo cyclization under basic (NaOMe/MeOH) or acidic (PPA)
conditions to provide intermediate (11h, R.sup.4a.dbd.H) or
compound (I-I, where R.sup.4a.dbd.R.sup.4). Alternatively, ester
(12a) may be hydrolyzed to prove acid (11 g). Methods described in
Scheme XI can be used to convert intermediate (11g) and compound
(I-I) to compounds of Formula (I-J).
[0087] Representative examples of compounds of Formula (I-I), (I-J)
or (I-K) that may be prepared by procedures described above in
Schemes XI and XII include:
3-(4-chlorophenyl)-2-(2-chlorophenyl)-7-(2,2,2-trifluoroethyl)-4,5,6,7-te-
trahydro-3H-imidazo[4,5-e][1,4]diazepin-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4-methyl-7-(2,2,2-trifluoroethyl)-4-
,5,6,7-tetrahydro-3H-imidazo[4,5-e][1,4]diazepin-8-one; and
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4,6,6-trimethyl-7-(2,2,2-trifluoroe-
thyl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-e][1,4]diazepin-8-one.
[0088] Scheme XII describes the preparation of compounds of formula
(I-L), (I-M), and (I-N) where A is nitrogen, B is carbon, and X is
S, SO, or SO.sub.2. ##STR17##
[0089] Compound (13a, see Andreichikov et al., J. Org. Chem. USSR
(Engl. Transl.), 23(4), 798-792 (1987)) is reacted with thioacetic
acid in a polar aprotic solvent (e.g, ether or THF) in the presence
of an amine base (e.g., triethylamine) at temperatures ranging from
0.degree. C. to 100.degree. C. to give intermediate (13b).
Condensation of (13b) with a substituted hydrazino derivative
(R.sup.0NHNH.sub.2) in the presence of an acid catalyst (e.g.,
sulfuric acid, acetic acid) in a solvent such as ethanol,
isopropanol, or toluene can provide the pyrazolyl intermediate
(13c). Removal of the thioacetyl protecting group with a reducing
agent such as LiBH.sub.4 in a polar solvent such as THF at
temperatures ranging from 0.degree. to 80.degree. C. can provide
the 4-mercapto-1H-pyrazole-3-carboxylic acid alkyl ester derivative
(13d). Compound (13d) can be reacted with an appropriately
substituted or unsubstituted .beta.-haloethyleneamine derivative
where the amino group is protected with a suitable group (i.e., Pg)
in the presence of a mild base (e.g, Na.sub.2CO.sub.3,
K.sub.2CO.sub.3). Deprotection of the amino group under standard
conditions can provide compounds such as (13f) which can be
cyclized in the presence of acid (e.g., PPA) or base (e.g.
NaOMe/MeOH) as described earlier. Alkylation of (13g) using
procedures analogous to those shown in Scheme IV can yield
compounds such as (I-L). Compound (I-L) can be converted to the
corresponding sulfoxide (I-M) or sulfone (I-N) using an oxidizing
agent such as m-chloroperbenzoic acid (m-CPBA) or an oxaziridine,
with the oxidation state of the sulfur atom dependent on the
reaction time.
[0090] Representative examples of compounds of Formula (I-L), (I-M)
and (I-N) that may be prepared by the procedures described above in
Scheme XIII include:
3-(4-chlorophenyl)-2-(2-chlorophenyl)-7-(2,2,2-trifluoroethyl)-6,7-dihydr-
o-2H,5H-4-thia-1,2,7-triaza-azulen-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4-oxo-7-(2,2,2-trifluoroethyl)-4,5,-
6,7-tetrahydro-2H-4.lamda..sup.4-thia-1,2,7-triaza-azulen-8-one;
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4,4-dioxo-7-(2,2,2-trifluoroethyl)--
4,5,6,7-tetrahydro-2H-4.lamda..sup.4-thia-1,2,7-triaza-azulen-8-one;
and
3-(4-chlorophenyl)-2-(2-chlorophenyl)-6,6-dimethyl-7-(2,2,2-trifluoroethy-
l)-6,7-dihydro-2H,5H-4-thia-1,2,7-triaza-azulen-8-one.
[0091] Scheme XIV describes the preparation of compounds of Formula
(I-O), (I-P), and (I-Q) where A is carbon, B is nitrogen, and X is
S, SO, or SO.sub.2. ##STR18##
[0092] Compound (11a) can be reacted with cysteamine or a
substituted 2-amino-propane-1-thiol derivative in a solvent such as
DMF in the presence of a base such as
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) to provide intermediate
(14a) which can then be cyclized under acidic (e.g., polyphosphoric
acid (PPA)) or basic (e.g., NaOMe/MeOH) conditions to yield
intermediate (14b, R.sup.4a.dbd.H) or compound (I-O,
R.sup.4a.dbd.R.sup.4). Alkylation of (14b) using procedures
analogous to those shown in Scheme V can yield compounds such as
(I-O). Compound (I-O) can be converted to the corresponding
sulfoxide (I-P) or sulfone (I-Q) using an oxidizing agent as
previously described.
[0093] Representative examples of compounds of Formula (I-O),
(I-P), and (I-Q) that may be prepared by the procedures described
above in Scheme XIV include:
2-(2-chlorophenyl)-3-(4-chlorophenyl)-7-(2,2,2-trifluoroethyl)-6,7-dihydr-
o-3H,5H-4-thia-1,3,7-triaza-azulen-8-one;
2-(2-chlorophenyl)-3-(4-chlorophenyl)-4-oxo-7-(2,2,2-trifluoroethyl)-4,5,-
6,7-tetrahydro-3H-4.lamda..sup.4-thia-1,3,7-triaza-azulen-8-one;
2-(2-chlorophenyl)-3-(4-chlorophenyl)-4,4-dioxo-7-(2,2,2-trifluoroethyl)--
4,5,6,7-tetrahydro-3H-4.lamda..sup.4-thia-1,3,7-triaza-azulen-8-one;
and
2-(2-chlorophenyl)-3-(4-chlorophenyl)-6,6-dimethyl-7-(2,2,2-trifluoroethy-
l)-6,7-dihydro-3H, 5H-4-thia-1,3,7-triaza-azulen-8-one.
[0094] Compounds of Formula (I-R) where A is carbon, B is nitrogen,
and X is C(R.sup.2b)(R.sup.2c) can be prepared as shown in Scheme
XV. ##STR19##
[0095] The imidazolyl intermediate (11a) can be converted into
intermediate (15e) by methods analogous to those described in
Scheme IV. Treatment of (15e) with a strong base (e.g., sodium
hydride) in an aprotic solvent (e.g., THF) can provide compounds of
formula (I-R).
[0096] Representative examples of compounds of Formula (I-R) that
may be prepared by the procedures described above in Scheme XIV
include:
2-(2-chlorophenyl)-1-(4-chlorophenyl)-5-(2,2-difluoropropyl)-1,6,7,8-tetr-
ahydro-5H-1,3,5-triaza-azulen-4-one;
2-(2-chlorophenyl)-1-(4-chlorophenyl)-5-(2,2,2-trifluoroethyl)-1,6,7,8-te-
trahydro-5H-1,3,5-triaza-azulen-4-one; and
2-(2-chlorophenyl)-1-(4-chlorophenyl)-8,8-dimethyl-5-(2,2,2-trifluoroethy-
l)-1,6,7,8-tetrahydro-5H-1,3,5-triaza-azulen-4-one.
[0097] Conventional methods and/or techniques of separation and
purification known to one of ordinary skill in the art can be used
to isolate the compounds of the present invention, as well as the
various intermediates related thereto. Such techniques will be
well-known to one of ordinary skill in the art and may include, for
example, all types of chromatography (high pressure liquid
chromatography (HPLC), column chromatography using common
adsorbents such as silica gel, and thin-layer chromatography),
recrystallization, and differential (i.e., liquid-liquid)
extraction techniques.
[0098] The compounds of the present invention may be isolated and
used per se or in the form of its pharmaceutically acceptable salt,
solvate and/or hydrate. The term "salts" refers to inorganic and
organic salts of a compound of the present invention. These salts
can be prepared in situ during the final isolation and purification
of a compound, or by separately reacting the compound or prodrug
with a suitable organic or inorganic acid or base and isolating the
salt thus formed. Representative salts include the hydrobromide,
hydrochloride, hydroiodide, sulfate, bisulfate, nitrate, acetate,
trifluoroacetate, oxalate, besylate, palmitiate, pamoate, malonate,
stearate, laurate, malate, borate, benzoate, lactate, phosphate,
hexafluorophosphate, benzene sulfonate, tosylate, formate, citrate,
maleate, fumarate, succinate, tartrate, naphthylate, mesylate,
glucoheptonate, lactobionate, and laurylsulphonate salts, and the
like. These may include cations based on the alkali and alkaline
earth metals, such as sodium, lithium, potassium, calcium,
magnesium, and the like, as well as non-toxic ammonium, quaternary
ammonium, and amine cations including, but not limited to,
ammonium, tetramethylammonium, tetraethylammonium, methylamine,
dimethylamine, trimethylamine, triethylamine, ethylamine, and the
like. See, e.g., Berge, et al., J. Pharm. Sci., 66, 1-19
(1977).
[0099] The term "prodrug" means a compound that is transformed in
vivo to yield a compound of Formula (I) or a pharmaceutically
acceptable salt, hydrate or solvate of the compound. The
transformation may occur by various mechanisms, such as through
hydrolysis in blood. A discussion of the use of prodrugs is
provided by T. Higuchi and W. Stella, "Pro-drugs as Novel Delivery
Systems," Vol. 14 of the A.C.S. Symposium Series, and in
Bioreversible Carriers in Drug Design, ed. Edward B. Roche,
American Pharmaceutical Association and Pergamon Press, 1987.
[0100] For example, if a compound of the present invention contains
a carboxylic acid functional group, a prodrug can comprise an ester
formed by the replacement of the hydrogen atom of the acid group
with a group such as (C.sub.1-C.sub.8)alkyl,
(C.sub.2-C.sub.12)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having
from 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having
from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to
6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7
carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to
8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9
carbon atoms, 1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10
carbon atoms, 3-phthalidyl, 4-crotonolactonyl,
gamma-butyrolacton-4-yl,
di-N,N-(C.sub.1-C.sub.2)alkylamino(C.sub.2-C.sub.3)alkyl (such as
.beta.-dimethylaminoethyl), carbamoyl-(C.sub.1-C.sub.2)alkyl,
N,N-di(C.sub.1-C.sub.2)alkylcarbamoyl-(C.sub.1-C.sub.2)alkyl and
piperidino-, pyrrolidino- or morpholino(C.sub.2-C.sub.3)alkyl.
[0101] Similarly, if a compound of the present invention contains
an alcohol functional group, a prodrug can be formed by the
replacement of the hydrogen atom of the alcohol group with a group
such as (C.sub.1-C.sub.6)alkanoyloxymethyl,
1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
1-methyl-1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
(C.sub.1-C.sub.6)alkoxycarbonyloxymethyl,
N-(C.sub.1-C.sub.6)alkoxycarbonylaminomethyl, succinoyl,
(C.sub.1-C.sub.6)alkanoyl, .alpha.-amino(C.sub.1-C.sub.4)alkanoyl,
arylacyl and .alpha.-aminoacyl, or
.alpha.-aminoacyl-.alpha.-aminoacyl, where each .alpha.-aminoacyl
group is independently selected from the naturally occurring
L-amino acids, P(O)(OH).sub.2, P(O)(O(C.sub.1-C.sub.6)alkyl).sub.2
or glycosyl (the radical resulting from the removal of a
hydroxy-group of the hemiacetal form of a carbohydrate).
[0102] If a compound of the present invention incorporates an amine
functional group, a prodrug can be formed by the replacement of a
hydrogen atom in the amine group with a group such as R-carbonyl,
RO-carbonyl, NRR'-carbonyl where R and R' are each independently
(C.sub.1-C.sub.10)alkyl, (C.sub.3-C.sub.7)cycloalkyl, benzyl, or
R-carbonyl is a natural .alpha.-aminoacyl or natural
.alpha.-aminoacyl-natural .alpha.-aminoacyl, --C(OH)C(O)OY' wherein
Y' is H, (C.sub.1-C.sub.6)alkyl or benzyl, --C(OY.sub.0)Y.sub.1
wherein Y.sub.0 is (C.sub.1-C.sub.4) alkyl and Y.sub.1 is
(C.sub.1-C.sub.6)alkyl, carboxy(C.sub.1-C.sub.6)alkyl,
amino(C.sub.1-C.sub.4)alkyl or mono-N- or
di-N,N-(C.sub.1-C.sub.6)alkylaminoalkyl, --C(Y.sub.2)Y.sub.3
wherein Y.sub.2 is H or methyl and Y.sub.3 is mono-N-- or
di-N,N-(C.sub.1-C.sub.6)alkylamino, morpholino, piperidin-1-yl or
pyrrolidin-1-yl.
[0103] The compounds of the present invention may contain
asymmetric or chiral centers, and, therefore, exist in different
stereoisomeric forms. It is intended that all stereoisomeric forms
of the compounds of the present invention as well as mixtures
thereof, including racemic mixtures, form part of the present
invention. In addition, the present invention embraces all
geometric and positional isomers. For example, if a compound of the
present invention incorporates a double bond or a fused ring, both
the cis- and trans-forms, as well as mixtures, are embraced within
the scope of the invention.
[0104] Diastereomeric mixtures can be separated into their
individual diastereoisomers on the basis of their physical chemical
differences by methods well known to those skilled in the art, such
as by chromatography and/or fractional crystallization. Enantiomers
can be separated by converting the enantiomeric mixture into a
diastereomeric mixture by reaction with an appropriate optically
active compound (e.g., chiral auxiliary such as a chiral alcohol or
Mosher's acid chloride), separating the diastereoisomers and
converting (e.g., hydrolyzing) the individual diastereoisomers to
the corresponding pure enantiomers. Also, some of the compounds of
the present invention may be atropisomers (e.g., substituted
biaryls) and are considered as part of this invention. Enantiomers
can also be separated by use of a chiral HPLC column.
[0105] The compounds of the present invention may exist in
unsolvated as well as solvated forms with pharmaceutically
acceptable solvents such as water, ethanol, and the like, and it is
intended that the invention embrace both solvated and unsolvated
forms.
[0106] It is also possible that the intermediates and compounds of
the present invention may exist in different tautomeric forms, and
all such forms are embraced within the scope of the invention. The
term "tautomer" or "tautomeric form" refers to structural isomers
of different energies which are interconvertible via a low energy
barrier. For example, proton tautomers (also known as prototropic
tautomers) include interconversions via migration of a proton, such
as keto-enol and imine-enamine isomerizations. A specific example
of a proton tautomer is the imidazole moiety where the proton may
migrate between the two ring nitrogens. Valence tautomers include
interconversions by reorganization of some of the bonding
electrons.
[0107] The present invention also embraces isotopically-labeled
compounds of the present invention (including intermediates) which
are identical to those recited herein, but for the fact that one or
more atoms are replaced by an atom having an atomic mass or mass
number different from the atomic mass or mass number usually found
in nature. Examples of isotopes that can be incorporated into the
intermediates or compounds of the invention include isotopes of
hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine,
iodine, and chlorine, such as .sup.2H, .sup.3H, .sup.11C, .sup.13C
.sup.14C, .sup.13N, .sup.15N, .sup.15O, .sup.17O, .sup.18O,
.sup.31P, .sup.32P, .sup.35S, .sup.18F, .sup.123I, .sup.125I and
.sup.36Cl, respectively.
[0108] Certain isotopically-labeled compounds of the present
invention (e.g., those labeled with .sup.3H and .sup.14C) are
useful in compound and/or substrate tissue distribution assays.
Tritiated (i.e., .sup.3H) and carbon-14 (i.e., .sup.14C) isotopes
are particularly preferred for their ease of preparation and
detectability. Further, substitution with heavier isotopes such as
deuterium (i.e., .sup.2H) may afford certain therapeutic advantages
resulting from greater metabolic stability (e.g., increased in vivo
half-life or reduced dosage requirements) and hence may be
preferred in some circumstances. Positron emitting isotopes such as
.sup.15O, .sup.13N, .sup.11C, and .sup.18F are useful for positron
emission tomography (PET) studies to examine substrate receptor
occupancy. Isotopically labeled compounds of the present invention
can generally be prepared by following procedures analogous to
those disclosed in the Schemes and/or in the Examples herein below,
by substituting an isotopically labeled reagent for a
non-isotopically labeled reagent.
[0109] Compounds of the present invention are useful for treating
diseases, conditions and disorders modulated by cannabinoid
receptor antagonists; therefore, another embodiment of the present
invention is a pharmaceutical composition comprising a
therapeutically effective amount of a compound of the present
invention and a pharmaceutically acceptable excipient, diluent or
carrier.
[0110] A typical formulation is prepared by mixing a compound of
the present invention and a carrier, diluent or excipient. Suitable
carriers, diluents and excipients are well known to those skilled
in the art and include materials such as carbohydrates, waxes,
water soluble and/or swellable polymers, hydrophilic or hydrophobic
materials, gelatin, oils, solvents, water, and the like. The
particular carrier, diluent or excipient used will depend upon the
means and purpose for which the compound of the present invention
is being applied. Solvents are generally selected based on solvents
recognized by persons skilled in the art as safe (GRAS) to be
administered to a mammal. In general, safe solvents are non-toxic
aqueous solvents such as water and other non-toxic solvents that
are soluble or miscible in water. Suitable aqueous solvents include
water, ethanol, propylene glycol, polyethylene glycols (e.g.,
PEG400, PEG300), etc. and mixtures thereof. The formulations may
also include one or more buffers, stabilizing agents, surfactants,
wetting agents, lubricating agents, emulsifiers, suspending agents,
preservatives, antioxidants, opaquing agents, glidants, processing
aids, colorants, sweeteners, perfuming agents, flavoring agents and
other known additives to provide an elegant presentation of the
drug (i.e., a compound of the present invention or pharmaceutical
composition thereof) or aid in the manufacturing of the
pharmaceutical product (i.e., medicament).
[0111] The formulations may be prepared using conventional
dissolution and mixing procedures. For example, the bulk drug
substance (i.e., compound of the present invention or stabilized
form of the compound (e.g., complex with a cyclodextrin derivative
or other known complexation agent)) is dissolved in a suitable
solvent in the presence of one or more of the excipients described
above. The dissolution rate of poorly water-soluble compounds may
be enhanced by the use of a spray-dried dispersion, such as those
described by Takeuchi, H., et al. in "Enhancement of the
dissolution rate of a poorly water-soluble drug (tolbutamide) by a
spray-drying solvent depostion method and disintegrants" J. Pharm.
Pharmacol., 39, 769-773 (1987).
[0112] The compound of the present invention is typically
formulated into pharmaceutical dosage forms to provide an easily
controllable dosage of the drug and to give the patient an elegant
and easily handleable product.
[0113] The pharmaceutical composition (or formulation) for
application may be packaged in a variety of ways depending upon the
method used for administering the drug. Generally, an article for
distribution includes a container having deposited therein the
pharmaceutical formulation in an appropriate form. Suitable
containers are well-known to those skilled in the art and include
materials such as bottles (plastic and glass), sachets, ampoules,
plastic bags, metal cylinders, and the like. The container may also
include a tamper-proof assemblage to prevent indiscreet access to
the contents of the package. In addition, the container has
deposited thereon a label that describes the contents of the
container. The label may also include appropriate warnings.
[0114] The present invention further provides a method of treating
diseases, conditions and/or disorders modulated by cannabinoid
receptor antagonists in an animal that includes administering to an
animal in need of such treatment a therapeutically effective amount
of a compound of the present invention or a pharmaceutical
composition comprising an effective amount of a compound of the
present invention and a pharmaceutically acceptable excipient,
diluent, or carrier. The method is particularly useful for treating
diseases, conditions and/or disorders modulated by cannabinoid
receptor (in particular, CB1 receptor) antagonists.
[0115] Preliminary investigations have indicated that the following
diseases, conditions, and/or disorders are modulated by cannabinoid
receptor antagonists: eating disorders (e.g., binge eating
disorder, anorexia, and bulimia), weight loss or control (e.g.,
reduction in calorie or food intake, and/or appetite suppression),
obesity, depression, atypical depression, bipolar disorders,
psychoses, schizophrenia, behavioral addictions, suppression of
reward-related behaviors (e.g., conditioned place avoidance, such
as suppression of cocaine- and morphine-induced conditioned place
preference), substance abuse, addictive disorders, impulsivity,
alcoholism (e.g., alcohol abuse, addiction and/or dependence
including treatment for abstinence, craving reduction and relapse
prevention of alcohol intake), tobacco abuse (e.g., smoking
addiction, cessation and/or dependence including treatment for
craving reduction and relapse prevention of tobacco smoking),
dementia (including memory loss, Alzheimer's disease, dementia of
aging, vascular dementia, mild cognitive impairment, age-related
cognitive decline, and mild neurocognitive disorder), sexual
dysfunction in males (e.g., erectile difficulty), seizure
disorders, epilepsy, inflammation, gastrointestinal disorders
(e.g., dysfunction of gastrointestinal motility or intestinal
propulsion), attention deficit disorder (ADD including attention
deficit hyperactivity disorder (ADHD)), Parkinson's disease, and
type II diabetes.
[0116] Accordingly, the compounds of the present invention
described herein are useful in treating diseases, conditions, or
disorders that are modulated by cannabinoid receptor antagonists.
Consequently, the compounds of the present invention (including the
compositions and processes used therein) may be used in the
manufacture of a medicament for the therapeutic applications
described herein.
[0117] Other diseases, conditions and/or disorders for which
cannabinoid receptor antagonists may be effective include:
premenstrual syndrome or late luteal phase syndrome, migraines,
panic disorder, anxiety, post-traumatic syndrome, social phobia,
cognitive impairment in non-demented individuals, non-amnestic mild
cognitive impairment, post operative cognitive decline, disorders
associated with impulsive behaviours (such as, disruptive behaviour
disorders (e.g., anxiety/depression, executive function
improvement, tic disorders, conduct disorder and/or oppositional
defiant disorder), adult personality disorders (e.g., borderline
personality disorder and antisocial personality disorder), diseases
associated with impulsive behaviours (e.g., substance abuse,
paraphilias and self-mutilation), and impulse control disorders
(e.g., intermittene explosive disorder, kleptomania, pyromania,
pathological gambling, and trichotillomania)), obsessive compulsive
disorder, chronic fatigue syndrome, sexual dysfunction in males
(e.g., premature ejaculation), sexual dysfunction in females,
disorders of sleep (e.g., sleep apnea), autism, mutism,
neurodengenerative movement disorders, spinal cord injury, damage
of the central nervous system (e.g., trauma), stroke,
neurodegenerative diseases or toxic or infective CNS diseases
(e.g., encephalitis or meningitis), cardiovascular disorders (e.g.,
thrombosis), and diabetes.
[0118] The compounds of the present invention can be administered
to a patient at dosage levels in the range of from about 0.7 mg to
about 7,000 mg per day. For a normal adult human having a body
weight of about 70 kg, a dosage in the range of from about 0.01 mg
to about 100 mg per kilogram body weight is typically sufficient.
However, some variability in the general dosage range may be
required depending upon the age and weight of the subject being
treated, the intended route of administration, the particular
compound being administered and the like. The determination of
dosage ranges and optimal dosages for a particular patient is well
within the ability of one of ordinary skill in the art having the
benefit of the instant disclosure. It is also noted that the
compounds of the present invention can be used in sustained
release, controlled release, and delayed release formulations,
which forms are also well known to one of ordinary skill in the
art.
[0119] The compounds of this invention may also be used in
conjunction with other pharmaceutical agents for the treatment of
the diseases, conditions and/or disorders described herein.
Therefore, methods of treatment that include administering
compounds of the present invention in combination with other
pharmaceutical agents are also provided. Suitable pharmaceutical
agents that may be used in combination with the compounds of the
present invention include anti-obesity agents such as
apolipoprotein-B secretion/microsomal triglyceride transfer protein
(apo-B/MTP) inhibitors, 11.beta.-hydroxy steroid dehydrogenase-1
(11.beta.-HSD type 1) inhibitors, peptide YY.sub.3-36 or analogs
thereof, MCR-4 agonists, cholecystokinin-A (CCK-A) agonists,
monoamine reuptake inhibitors (such as sibutramine),
sympathomimetic agents, .beta..sub.3 adrenergic receptor agonists,
dopamine agonists (such as bromocriptine), melanocyte-stimulating
hormone receptor analogs, 5HT2c agonists, melanin concentrating
hormone antagonists, leptin (the OB protein), leptin analogs,
leptin receptor agonists, galanin antagonists, lipase inhibitors
(such as tetrahydrolipstatin, i.e. orlistat), anorectic agents
(such as a bombesin agonist), Neuropeptide-Y antagonists,
thyromimetic agents, dehydroepiandrosterone or an analog thereof,
glucocorticoid receptor agonists or antagonists, orexin receptor
antagonists, glucagon-like peptide-1 receptor agonists, ciliary
neurotrophic factors (such as Axokine.TM. available from Regeneron
Pharmaceuticals, Inc., Tarrytown, N.Y. and Procter & Gamble
Company, Cincinnati, Ohio), human agouti-related proteins (AGRP),
ghrelin receptor antagonists, histamine 3 receptor antagonists or
inverse agonists, neuromedin U receptor agonists and the like.
Other anti-obesity agents, including the preferred agents set forth
hereinbelow, are well known, or will be readily apparent in light
of the instant disclosure, to one of ordinary skill in the art.
[0120] Especially preferred are anti-obesity agents selected from
the group consisting of orlistat, sibutramine, bromocriptine,
ephedrine, leptin, pseudoephedrine and peptide YY.sub.3-36 or an
analog thereof. Preferably, compounds of the present invention and
combination therapies are administered in conjunction with exercise
and a sensible diet.
[0121] Representative anti-obesity agents for use in the
combinations, pharmaceutical compositions, and methods of the
invention can be prepared using methods known to one of ordinary
skill in the art, for example, sibutramine can be prepared as
described in U.S. Pat. No. 4,929,629; bromocriptine can be prepared
as described in U.S. Pat. Nos. 3,752,814 and 3,752,888; orlistat
can be prepared as described in U.S. Pat. Nos. 5,274,143;
5,420,305; 5,540,917; and 5,643,874; and PYY.sub.3-36 (including
analogs) can be prepared as described in US Publication No.
2002/0141985 and WO 03/027637. All of the above recited references
are incorporated herein by reference.
[0122] Other suitable pharmaceutical agents that may be
administered in combination with the compounds of the present
invention include agents designed to treat tobacco abuse (e.g.,
nicotine receptor partial agonists, bupropion hypochloride (also
known under the tradename Zyban.TM.) and nicotine replacement
therapies), agents to treat erectile dysfunction (e.g.,
dopaminergic agents, such as apomorphine), ADD/ADHD agents (e.g.,
Ritalin.TM., Strattera.TM., Concerta.TM. and Adderall.TM.), and
agents to treat alcoholism, such as opioid antagonists (e.g.,
naltrexone (also known under the tradename ReVia.TM.) and
nalmefene), disulfiram (also known under the tradename
Antabuse.TM.), and acamprosate (also known under the tradename
Campral.TM.)). In addition, agents for reducing alcohol withdrawal
symptoms may also be co-administered, such as benzodiazepines,
beta-blockers, clonidine, carbamazepine, pregabalin, and gabapentin
(Neurontin.TM.). Treatment for alcoholism is preferably
administered in combination with behavioral therapy including such
components as motivational enhancement therapy, cognitive
behavioral therapy, and referral to self-help groups, including
Alcohol Anonymous (AA).
[0123] Other pharmaceutical agents that may be useful include
antihypertensive agents; anti-inflammatory agents (e.g., COX-2
inhibitors); antidepressants (e.g., fluoxetine hydrochloride
(Prozac.TM.)); cognitive improvement agents (e.g., donepezil
hydrochloride (Aircept.TM.) and other acetylcholinesterase
inhibitors); neuroprotective agents (e.g., memantine);
antipsychotic medications (e.g., ziprasidone (Geodon.TM.),
risperidone (Risperdal.TM.), and olanzapine (Zyprexa.TM.)); insulin
and insulin analogs (e.g., LysPro insulin); GLP-1 (7-37)
(insulinotropin) and GLP-1 (7-36)-NH.sub.2; sulfonylureas and
analogs thereof: chlorpropamide, glibenclamide, tolbutamide,
tolazamide, acetohexamide, Glypizide.RTM., glimepiride,
repaglinide, meglitinide; biguanides: mefformin, phenformin,
buformin; .alpha.2-antagonists and imidazolines: midaglizole,
isaglidole, deriglidole, idazoxan, efaroxan, fluparoxan; other
insulin secretagogues: linogliride, A-4166; glitazones:
ciglitazone, Actos.RTM. (pioglitazone), englitazone, troglitazone,
darglitazone, Avandia.RTM. (BRL49653); fatty acid oxidation
inhibitors: clomoxir, etomoxir; .alpha.-glucosidase inhibitors:
acarbose, miglitol, emiglitate, voglibose, MDL-25,637, camiglibose,
MDL-73,945; .beta.-agonists: BRL 35135, BRL 37344, RO 16-8714, ICI
D7114, CL 316,243; phosphodiesterase inhibitors: L-386,398;
lipid-lowering agents: benfluorex: fenfluramine; vanadate and
vanadium complexes (e.g., Naglivan.RTM.) and peroxovanadium
complexes; amylin antagonists; glucagon antagonists;
gluconeogenesis inhibitors; somatostatin analogs; antilipolytic
agents: nicotinic acid, acipimox, WAG 994, pramlintide
(Symlin.TM.), AC 2993, nateglinide, aldose reductase inhibitors
(e.g., zopolrestat), glycogen phosphorylase inhibitors, sorbitol
dehydrogenase inhibitors, sodium-hydrogen exchanger type 1 (NHE-1)
inhibitors and/or cholesterol biosynthesis inhibitors or
cholesterol absorption inhibitors, especially a HMG-COA reductase
inhibitor (e.g., atorvastatin or the hemicalcium salt thereof), or
a HMG-CoA synthase inhibitor, or a HMG-CoA reductase or synthase
gene expression inhibitor, a CETP inhibitor, a bile acid
sequesterant, a fibrate, an ACAT inhibitor, a squalene synthetase
inhibitor, an anti-oxidant or niacin. The compounds of the present
invention may also be administered in combination with a naturally
occurring compound that acts to lower plasma cholesterol levels.
Such naturally occurring compounds are commonly called
nutraceuticals and include, for example, garlic extract, Hoodia
plant extracts, and niacin.
[0124] The dosage of the additional pharmaceutical agent will also
be generally dependent upon a number of factors including the
health of the subject being treated, the extent of treatment
desired, the nature and kind of concurrent therapy, if any, and the
frequency of treatment and the nature of the effect desired. In
general, the dosage range of an anti-obesity agent is in the range
of from about 0.001 mg to about 100 mg per kilogram body weight of
the individual per day, preferably from about 0.1 mg to about 10 mg
per kilogram body weight of the individual per day. However, some
variability in the general dosage range may also be required
depending upon the age and weight of the subject being treated, the
intended route of administration, the particular anti-obesity agent
being administered and the like. The determination of dosage ranges
and optimal dosages for a particular patient is also well within
the ability of one of ordinary skill in the art having the benefit
of the instant disclosure.
[0125] According to the methods of the invention, a compound of the
present invention or a combination of a compound of the present
invention and at least one additional pharmaceutical agent is
administered to a subject in need of such treatment, preferably in
the form of a pharmaceutical composition. In the combination aspect
of the invention, the compound of the present invention and at
least one other pharmaceutical agent may be administered either
separately or in the pharmaceutical composition comprising both. It
is generally preferred that such administration be oral. However,
if the subject being treated is unable to swallow, or oral
administration is otherwise impaired or undesirable, parenteral or
transdermal administration may be appropriate.
[0126] According to the methods of the invention, when a
combination of a compound of the present invention and at least one
other pharmaceutical agent are administered together, such
administration can be sequential in time or simultaneous with the
simultaneous method being generally preferred. For sequential
administration, a compound of the present invention and the
additional pharmaceutical agent can be administered in any order.
It is generally preferred that such administration be oral. It is
especially preferred that such administration be oral and
simultaneous. When a compound of the present invention and the
additional pharmaceutical agent are administered sequentially, the
administration of each can be by the same or by different
methods.
[0127] According to the methods of the invention, a compound of the
present invention or a combination of a compound of the present
invention and at least one additional pharmaceutical agent
(referred to herein as a "combination") is preferably administered
in the form of a pharmaceutical composition. Accordingly, a
compound of the present invention or a combination can be
administered to a patient separately or together in any
conventional oral, rectal, transdermal, parenteral, (for example,
intravenous, intramuscular, or subcutaneous) intracisternal,
intravaginal, intraperitoneal, intravesical, local (for example,
powder, ointment or drop), or buccal, or nasal, dosage form.
[0128] Compositions suitable for parenteral injection generally
include pharmaceutically acceptable sterile aqueous or nonaqueous
solutions, dispersions, suspensions, or emulsions, and sterile
powders for reconstitution into sterile injectable solutions or
dispersions. Examples of suitable aqueous and nonaqueous carriers,
diluents, solvents, or vehicles include water, ethanol, polyols
(propylene glycol, polyethylene glycol, glycerol, and the like),
suitable mixtures thereof, vegetable oils (such as olive oil) and
injectable organic esters such as ethyl oleate. Proper fluidity can
be maintained, for example, by the use of a coating such as
lecithin, by the maintenance of the required particle size in the
case of dispersions, and by the use of surfactants.
[0129] These compositions may also contain adjuvants such as
preserving, wetting, emulsifying, and dispersing agents. Prevention
of microorganism contamination of the compositions can be
accomplished with various antibacterial and antifungal agents, for
example, parabens, chlorobutanol, phenol, sorbic acid, and the
like. It may also be desirable to include isotonic agents, for
example, sugars, sodium chloride, and the like. Prolonged
absorption of injectable pharmaceutical compositions can be brought
about by the use of agents capable of delaying absorption, for
example, aluminum monostearate and gelatin.
[0130] Solid dosage forms for oral administration include capsules,
tablets, powders, and granules. In such solid dosage forms, a
compound of the present invention or a combination is admixed with
at least one inert customary pharmaceutical excipient (or carrier)
such as sodium citrate or dicalcium phosphate or (a) fillers or
extenders (e.g., starches, lactose, sucrose, mannitol, silicic acid
and the like); (b) binders (e.g., carboxymethylcellulose,
alginates, gelatin, polyvinylpyrrolidone, sucrose, acacia and the
like); (c) humectants (e.g., glycerol and the like); (d)
disintegrating agents (e.g., agar-agar, calcium carbonate, potato
or tapioca starch, alginic acid, certain complex silicates, sodium
carbonate and the like); (e) solution retarders (e.g., paraffin and
the like); (f) absorption accelerators (e.g., quaternary ammonium
compounds and the like); (g) wetting agents (e.g., cetyl alcohol,
glycerol monostearate and the like); (h) adsorbents (e.g., kaolin,
bentonite and the like); and/or (i) lubricants (e.g., talc, calcium
stearate, magnesium stearate, solid polyethylene glycols, sodium
lauryl sulfate and the like). In the case of capsules and tablets,
the dosage forms may also comprise buffering agents.
[0131] Solid compositions of a similar type may also be used as
fillers in soft or hard filled gelatin capsules using such
excipients as lactose or milk sugar, as well as high molecular
weight polyethylene glycols, and the like.
[0132] Solid dosage forms such as tablets, dragees, capsules, and
granules can be prepared with coatings and shells, such as enteric
coatings and others well known in the art. They may also contain
opacifying agents, and can also be of such composition that they
release the compound of the present invention and/or the additional
pharmaceutical agent in a delayed manner. Examples of embedding
compositions that can be used are polymeric substances and waxes.
The drug can also be in micro-encapsulated form, if appropriate,
with one or more of the above-mentioned excipients.
[0133] Liquid dosage forms for oral administration include
pharmaceutically acceptable emulsions, solutions, suspensions,
syrups, and elixirs. In addition to the compound of the present
invention or the combination, the liquid dosage form may contain
inert diluents commonly used in the art, such as water or other
solvents, solubilizing agents and emulsifiers, as for example,
ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate,
benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene
glycol, dimethylformamide, oils (e.g., cottonseed oil, groundnut
oil, corn germ oil, olive oil, castor oil, sesame seed oil and the
like), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols
and fatty acid esters of sorbitan, or mixtures of these substances,
and the like.
[0134] Besides such inert diluents, the composition can also
include adjuvants, such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, and perfuming agents.
[0135] Suspensions, in addition to the compound of the present
invention or the combination, may further comprise suspending
agents, e.g., ethoxylated isostearyl alcohols, polyoxyethylene
sorbitol and sorbitan esters, microcrystalline cellulose, aluminum
metahydroxide, bentonite, agar-agar, and tragacanth, or mixtures of
these substances, and the like.
[0136] Compositions for rectal or vaginal administration preferably
comprise suppositories, which can be prepared by mixing a compound
of the present invention or a combination with suitable
non-irritating excipients or carriers, such as cocoa butter,
polyethylene glycol or a suppository wax which are solid at
ordinary room temperature but liquid at body temperature and
therefore melt in the rectum or vaginal cavity thereby releasing
the active component(s).
[0137] Dosage forms for topical administration of the compounds of
the present invention and combinations of the compounds of the
present invention with anti-obesity agents may comprise ointments,
powders, sprays and inhalants. The drugs are admixed under sterile
condition with a pharmaceutically acceptable carrier, and any
preservatives, buffers, or propellants that may be required.
Ophthalmic formulations, eye ointments, powders, and solutions are
also intended to be included within the scope of the present
invention.
[0138] The following paragraphs describe exemplary formulations,
dosages, etc. useful for non-human animals. The administration of
the compounds of the present invention and combinations of the
compounds of the present invention with anti-obesity agents can be
effected orally or non-orally (e.g., by injection).
[0139] An amount of a compound of the present invention or
combination of a compound of the present invention with an
anti-obesity agent is administered such that an effective dose is
received. Generally, a daily dose that is administered orally to an
animal is between about 0.01 and about 1,000 mg/kg of body weight,
preferably between about 0.01 and about 300 mg/kg of body
weight.
[0140] Conveniently, a compound of the present invention (or
combination) can be carried in the drinking water so that a
therapeutic dosage of the compound is ingested with the daily water
supply. The compound can be directly metered into drinking water,
preferably in the form of a liquid, water-soluble concentrate (such
as an aqueous solution of a water-soluble salt).
[0141] Conveniently, a compound of the present invention (or
combination) can also be added directly to the feed, as such, or in
the form of an animal feed supplement, also referred to as a premix
or concentrate. A premix or concentrate of the compound in a
carrier is more commonly employed for the inclusion of the agent in
the feed. Suitable carriers are liquid or solid, as desired, such
as water, various meals such as alfalfa meal, soybean meal,
cottonseed oil meal, linseed oil meal, corncob meal and corn meal,
molasses, urea, bone meal, and mineral mixes such as are commonly
employed in poultry feeds. A particularly effective carrier is the
respective animal feed itself; that is, a small portion of such
feed. The carrier facilitates uniform distribution of the compound
in the finished feed with which the premix is blended. Preferably,
the compound is thoroughly blended into the premix and,
subsequently, the feed. In this respect, the compound may be
dispersed or dissolved in a suitable oily vehicle such as soybean
oil, corn oil, cottonseed oil, and the like, or in a volatile
organic solvent and then blended with the carrier. It will be
appreciated that the proportions of compound in the concentrate are
capable of wide variation since the amount of the compound in the
finished feed may be adjusted by blending the appropriate
proportion of premix with the feed to obtain a desired level of
compound.
[0142] High potency concentrates may be blended by the feed
manufacturer with proteinaceous carrier such as soybean oil meal
and other meals, as described above, to produce concentrated
supplements, which are suitable for direct feeding to animals. In
such instances, the animals are permitted to consume the usual
diet. Alternatively, such concentrated supplements may be added
directly to the feed to produce a nutritionally balanced, finished
feed containing a therapeutically effective level of a compound of
the present invention. The mixtures are thoroughly blended by
standard procedures, such as in a twin shell blender, to ensure
homogeneity.
[0143] If the supplement is used as a top dressing for the feed, it
likewise helps to ensure uniformity of distribution of the compound
across the top of the dressed feed.
[0144] Drinking water and feed effective for increasing lean meat
deposition and for improving lean meat to fat ratio are generally
prepared by mixing a compound of the present invention with a
sufficient amount of animal feed to provide from about 10.sup.-3 to
about 500 ppm of the compound in the feed or water.
[0145] The preferred medicated swine, cattle, sheep and goat feed
generally contain from about 1 to about 400 grams of a compound of
the present invention (or combination) per ton of feed, the optimum
amount for these animals usually being about 50 to about 300 grams
per ton of feed.
[0146] The preferred poultry and domestic pet feeds usually contain
about 1 to about 400 grams and preferably about 10 to about 400
grams of a compound of the present invention (or combination) per
ton of feed.
[0147] For parenteral administration in animals, the compounds of
the present invention (or combination) may be prepared in the form
of a paste or a pellet and administered as an implant, usually
under the skin of the head or ear of the animal in which increase
in lean meat deposition and improvement in lean meat to fat ratio
is sought.
[0148] In general, parenteral administration involves injection of
a sufficient amount of a compound of the present invention (or
combination) to provide the animal with about 0.01 to about 20
mg/kg/day of body weight of the drug. The preferred dosage for
poultry, swine, cattle, sheep, goats and domestic pets is in the
range of from about 0.05 to about 10 mg/kg/day of body weight of
drug.
[0149] Paste formulations can be prepared by dispersing the drug in
a pharmaceutically acceptable oil such as peanut oil, sesame oil,
corn oil or the like.
[0150] Pellets containing an effective amount of a compound of the
present invention, pharmaceutical composition, or combination can
be prepared by admixing a compound of the present invention or
combination with a diluent such as carbowax, carnuba wax, and the
like, and a lubricant, such as magnesium or calcium stearate, can
be added to improve the pelleting process.
[0151] It is, of course, recognized that more than one pellet may
be administered to an animal to achieve the desired dose level
which will provide the increase in lean meat deposition and
improvement in lean meat to fat ratio desired. Moreover, implants
may also be made periodically during the animal treatment period in
order to maintain the proper drug level in the animal's body.
[0152] The present invention has several advantageous veterinary
features. For the pet owner or veterinarian who wishes to increase
leanness and/or trim unwanted fat from pet animals, the instant
invention provides the means by which this may be accomplished. For
poultry, beef and swine breeders, utilization of the method of the
present invention yields leaner animals that command higher sale
prices from the meat industry.
[0153] Embodiments of the present invention are illustrated by the
following Examples. It is to be understood, however, that the
embodiments of the invention are not limited to the specific
details of these Examples, as other variations thereof will be
known, or apparent in light of the instant disclosure, to one of
ordinary skill in the art.
EXAMPLES
General Experimental Procedures
[0154] NMR spectra were recorded on a Varian Unity.TM. 400
(available from Varian Inc., Palo Alto, Calif.) at room temperature
at 400 MHz for proton. Chemical shifts are expressed in parts per
million (6) relative to residual solvent as an internal reference.
The peak shapes are denoted as follows: s, singlet; d, doublet; t,
triplet; q, quartet; m, multiplet; bs, broad singlet; 2s, two
singlets. Atmospheric pressure chemical ionization mass spectra
(APCI) were obtained on a Fisons.TM. Platform II Spectrometer
(carrier gas: acetonitrile: available from Micromass Ltd,
Manchester, UK). Chemical ionization mass spectra (CI) were
obtained on a Hewlett-Packard.TM. 5989 instrument (ammonia
ionization, PBMS: available from Hewlett-Packard Company, Palo
Alto, Calif.). Electrospray ionization mass spectra (ES) were
obtained on a Waters.TM. ZMD instrument (carrier gas: acetonitrile:
available from Waters Corp., Milford, Mass.). Where the intensity
of chlorine or bromine-containing ions are described, the expected
intensity ratio was observed (approximately 3:1 for
.sup.35Cl/.sup.37Cl-containing ions and 1:1 for
.sup.79Br/.sup.81Br-containing ions) and the intensity of only the
lower mass ion is given. In some cases only representative .sup.1H
NMR peaks are given. MS peaks are reported for all examples.
Optical rotations were determined on a PerkinElmer.TM. 241
polarimeter (available from PerkinElmer Inc., Wellesley, Mass.)
using the sodium D line (.lamda.=589 nm) at the indicated
temperature and are reported as follows [.alpha.].sub.D.sup.temp,
concentration (c=g/100 ml), and solvent.
[0155] Column chromatography was performed with either Baker.TM.
silica gel (40 .mu.m; J.T. Baker, Phillipsburg, N.J.) or Silica Gel
50 (EM Sciences.TM., Gibbstown, N.J.) in glass columns or in Flash
40 Biotage.TM. columns (ISC, Inc., Shelton, Conn.) under low
nitrogen pressure.
[0156] Unless specified otherwise, starting materials are generally
available from commercial sources such as Aldrich Chemicals Co.
(Milwaukee, Wis.), Lancaster Synthesis, Inc. (Windham, N.H.), Acros
Organics (Fairlawn, N.J.), Maybridge Chemical Company, Ltd.
(Cornwall, England), Tyger Scientific (Princeton, N.J.), and
AstraZeneca Pharmaceuticals (London, England).
[0157] The 1,5-disubstituted 1H-pyrazole-3-carboxylic acid ester
starting materials were prepared using procedures analogous to
those described in U.S. Pat. No. 5,624,941 (Example No. 1) for the
preparation of methyl
1-(2,4-dichlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylate.
[0158] The following section provides representative examples of
useful intermediates that may be used in the synthesis of compounds
of the present invention.
Intermediates
Preparation of Intermediate
4-Bromo-5-(4-chlorophenyl)-1-(2-chlorophenyl)-1H-pyrazole-3-carboxylic
Acid Ethyl Ester (I-1a)
[0159] ##STR20##
[0160] Bromine (15 ml, 294 mmol) was added in one portion to a
cooled (ice/water bath) stirred solution of
5-(4-chlorophenyl)-1-(2-chlorophenyl)-1H-pyrazole-3-carboxylic acid
ethyl ester (26.6 g, 73.6 mmol) in acetic acid (300 ml). After 45
minutes, the reaction was concentrated in vacuo, the solids
slurried in diethyl ether (100 ml), filtered and dried in vacuo to
afford the title compound (I-1a) as a light-yellow colored solid,
29.6 g.
Preparation of Intermediate
5-(4-Chlorophenyl)-1-(2-chlorophenyl)-4-vinyl-1H-pyrazole-3-carboxylic
Acid Ethyl Ester (I-1b)
[0161] ##STR21##
[0162] A solution of
4-bromo-5-(4-chlorophenyl)-1-(2-chlorophenyl)-1H-pyrazole-3-carboxylic
acid ethyl ester I-1a (5.2 g, 11.9 mmol), tributylvinyltin (7.0 ml,
23.8 mmol) and tetrakistriphenylphosphine palladium (0.7 g, 0.6
mmol) in DMF (12 ml) was heated at 110.degree. C. for 18 hours. The
dark solution was cooled, partitioned between ethyl ether/water,
the organic layer washed with brine, dried (Na.sub.2SO.sub.4) and
concentrated in vacuo to afford a semi-solid. This semi-solid was
stirred with cyclohexanes (35 ml) and filtered to afford the title
compound (I-1 b) as a white solid, 3.0 g.
Preparation of Intermediate
5-(4-Chlorophenyl)-1-(2-chlorophenyl)-4-formyl-1H-pyrazole-3-carboxylic
Acid Ethyl Ester (I-1c)
[0163] ##STR22##
[0164] A solution of
5-(4-chlorophenyl)-1-(2-chlorophenyl)-4-vinyl-1H-pyrazole-3-carboxylic
acid ethyl ester I-1 b (2.9 g, 7.5 mmol), osmium tetroxide (8 mg,
0.08 mmol) and N-methylmorpholine-N-oxide (1.1 g, 8.2 mmol) in
dioxane (24 ml)/water (6 ml) was stirred at ambient temperature for
18 hours, then sodium periodate (16 g, 75 mmol) was added and
stirring was continued for 3.5 hours. The thick slurry was diluted
with ethyl acetate (100 ml), filtered and solids washed 2.times.
with ethyl acetate. The combined filtrates were washed with water,
brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford
a solid mass. The solids were slurried in hot hexanes (30 ml),
cooled, filtered and dried in vacuo to afford the title compound
(I-1c) as a tan solid, 2.2 g.
Preparation of Intermediate
5-(4-Chlorophenyl)-1-(2-chlorophenyl)-4-hydroxy-1H-pyrazole-3-carboxylic
Acid Ethyl Ester (I-1d)
[0165] ##STR23##
[0166] To a stirred solution of
5-(4-chlorophenyl)-1-(2-chlorophenyl)-4-formyl-1H-pyrazole-3-carboxylic
acid ethyl ester I-1c (2.2 g, 5.6 mmol) in dichloromethane (22 ml)
was added m-chloroperbenzoic acid (2.9 g (50% purity), 8.4 mmol)
and the resulting slurry was stirred for 6 hours. The mixture was
diluted into ethyl ether, washed with half-saturated aqueous sodium
bicarbonate, water, brine, dried (Na.sub.2SO.sub.4) and
concentrated in vacuo to afford a yellow solid, 3.5 g. To a slurry
of this material in methanol (20 ml), was added triethylamine (1
ml) to produce a solution. After 45 minutes, the reaction was
concentrated in vacuo to afford a yellow solid. This material was
purified by silica gel chromatograpy (Combiflash instrument, 120 g
silica gel column, 5-25% gradient of ethyl acetate/hexanes to
afford the title compound (I-1d) as a yellow solid, 1.5 g.
[0167] The following describes an alternative procedure for the
preparation of Intermediate
5-(4-Chlorophenyl)-1-(2-chlorophenyl)-4-hydroxy-1H-pyrazole-3-carboxylic
Acid Ethyl Ester (I-1d).
Preparation of Intermediate 4-(4-Chlorophenyl)-3-oxo-butyric Acid
Ethyl Ester (I-2a)
[0168] ##STR24##
[0169] To a cooled (0.degree. C.), stirred solution of
2,2-dimethyl-1,3-doxane-4,6-dione (78.5 g, 0.54 mol) in
dichloromethane (200 ml) was added pyridine (105 ml) dropwise over
a 30-minute period. A solution of 4-chlorophenylacetyl chloride
(100 g, 0.53 mol) in dichloromethane (150 ml) was added dropwise.
The reaction mixture was stirred for 1 hour at 0.degree. C., then
cooling bath was removed, and stirring was continued an additional
2 hours. Reaction mixture was poured over 2N hydrochloric acid
(aq.)/ice, layers separated and the aqueous layer washed with
dichloromethane (2.times.150 ml). Combined organic layers were
washed with 2N hydrochloric acid (aq.) (2.times.150 ml), brine,
dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford a
solid.
[0170] The material obtained above was slurried in ethanol (1
liter), heated to reflux for 3 hours, then cooled and concentrated
in vacuo. The oily residue was fractionally-distilled under vacuum
to afford the title compound (I-1a) as a clear oil, 108 g.
Preparation of Intermediate
4-(4-Chlorophenyl)-2-[(2-chlorophenyl)-hydrazono]-3-oxobutyric Acid
Ethyl Ester (I-2b)
[0171] ##STR25##
[0172] A solution of sodium nitrite (3.4 g, 50.4 mmol) in water (15
ml) was added dropwise over an hour period to a cooled (0.degree.
C.), stirred solution of 2-chloroaniline (6.4 g, 50.4 mmol) in
acetic acid (50 ml)/water (7 ml). Then a solution of
4-(4-chloro-phenyl)-3-oxo-butyric acid ethyl ester (10 g, 42 mmol)
in acetic acid (30 ml) was added dropwise over a 30-minutes period
to produce an orange slurry (20 ml of water added to aid stirring).
After an additional hour, the mixture was filtered, solids washed
with water and air-dried. Solids slurried in ethanol (75 ml) for 30
minutes, filtered, solids washed with methanol and dried in vacuo
to afford the title compound (I-2b) as an orange solid, 11.0 g.
Preparation of
4-Bromo-4-(4-chlorophenyl)-2-[(2-chlorophenyl)-hydrazono]-3-oxobutyric
Acid Ethyl Ester (I-2c)
[0173] ##STR26##
[0174] A stirred slurry of
4-(4-chlorophenyl)-2-[(2-chlorophenyl)-hydrazono]-3-oxobutyric acid
ethyl ester I-2b (10.0 g, 26 mmol) and copper(II) bromide (13.4 g,
59.8 mmol) in ethyl acetate (100 ml)/chloroform (100 ml) was heated
at 60.degree. C. for 3 hours. Reaction mixture cooled and filtered
through diatomaceous earth washing with chloroform. The filtrate
was diluted with dichloromethane, washed with water, brine, dried
(Na.sub.2SO.sub.4) and concentrated in vacuo to afford the title
compound (I-2c) as a red oil, 12.1 g.
Preparation of intermediate
5-(4-Chlorophenyl)-1-(2-chlorophenyl)-4-hydroxy-1H-pyrazole-3-carboxylic
Acid Ethyl Ester (I-1d)
[0175] A mixture of
4-bromo-4-(4-chloro-phenyl)-2-[(2-chloro-phenyl)-hydrazono]-3-oxo-butyric
acid ethyl ester (12.1 g, 26 mmol) and sodium acetate (10.8 g, 130
mmol) in methanol (100 ml) was heated at reflux for 4 hours,
cooled, concentrated in vacuo and the residue partitioned between
ethyl acetate and water. The organic layer was washed with brine,
dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford a
solid. A slurry of this material in cyclohexane was heated to
reflux and allowed to stir at ambient temperature for 2 hours, and
filtered to afford the title compound (I-1d) as a yellow solid
(I-1d), 6.5 g.
Preparation of Intermediate
4-Allyloxy-5-(4-chlorophenyl)-1-(2-chlorophenyl)-1H-Pyrazole-3-carboxylic
Acid Ethyl Ester (I-1e)
[0176] ##STR27##
[0177] To a slurry of sodium hydride (39 mg of 60% in oil) in DMSO
(2.4 ml) was added
5-(4-chlorophenyl)-1-(2-chlorophenyl)-4-hydroxy-1H-pyrazole-3-carboxylic
acid ethyl ester I-1d (300 mg, 0.8 mmol) and the mixture was
agitated for 45 minutes. Allyl bromide (0.1 ml, 1.2 mmol) was added
and stirring was continued for 4.5 hours. The reaction solution was
diluted into ethyl acetate, washed with water (2.times.), brine,
dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford the
title compound (I-1e) as an orange oil, 340 mg. This material was
taken onto the next step without further purification.
Preparation of Intermediate
5-(4-Chlorophenyl)-1-(2-chlorophenyl)-4-(2-oxoethoxy)-1H-pyrazole-3-carbo-
xylic Acid Ethyl Ester (I-1)
[0178] ##STR28##
[0179] To a stirred solution of
4-allyloxy-5-(4-chlorophenyl)-1-(2-chlorophenyl)-1H-pyrazole-3-carboxylic
acid ethyl ester I-1e (337 mg, 0.8 mmol) osmium tetroxide (53
microliters of a 0.15 M solution in water) and
N-methylmorpholine-N-oxide (120 mg, 0.9 mmol) in dioxane (2.4
ml)/water (0.6 ml) was stirred at ambient temperature for 18 hours,
then sodium periodate (1.7 g, 8.1 mmol) was added and stirring was
continued for 3.5 hours. The thick slurry was diluted with ethyl
acetate (100 ml), filtered and solids washed 2.times. with ethyl
acetate. The combined filtrates were washed with water, brine,
dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford an
oil. Silica gel chromatography (25% ethyl acetate/hexanes) afforded
the title compound (I-1f) as a colorless foam, 130 mg.
Preparation of intermediate
5-(4-Chlorophenyl)-1-(2-chlorophenyl)-4-[2-(2,2,2-trifluoroethylamino)-et-
hoxy]-1H-pyrazole-3-carboxylic Acid Ethyl Ester (I-1q)
[0180] ##STR29##
[0181] A solution of
5-(4-chlorophenyl)-1-(2-chlorophenyl)-4-(2-oxoethoxy)-1H-pyrazole-3-carbo-
xylic acid ethyl ester I-1f (40 mg, 0.1 mmol),
2,2,2-trifluoroethylamine (14 mg, 0.14 mmol), sodium
triacetoxyborohydride (30 mg, 014 mmol) and acetic acid (6
microliters, 0.1 mmol) in 1,2-dichloroethane (0.5 ml) was stirred
for 18 hours. The reaction was partitioned between ethyl
acetate/saturated aqueous sodium bicarbonate, the organic layer was
dried (Na.sub.2SO.sub.4) and concentrated in vacuo to afford the
title compound (I-1g). This oil was taken on to the next step
without further purification.
Preparation of Intermediate
5-(4-Chlorophenyl)-1-(2-chlorophenyl)-4-[2-(2,2,2-trifluoroethylamino)-et-
hoxy]-1H-pyrazole-3-carboxylic Acid, Hydrochloride (I-1h)
[0182] ##STR30##
[0183] A solution of
5-(4-chlorophenyl)-1-(2-chlorophenyl)-4-[2-(2,2,2-trifluoroethylamino)-et-
hoxy]-1H-pyrazole-3-carboxylic acid ethyl ester I-1g (48 mg, 0.1
mmol) and 6 N aqueous KOH (0.1 ml) in ethanol (1 ml) was heated at
50.degree. C. for 2 hours. The reaction solution was cooled,
acidified with concentrated aqueous hydrochloric acid, and
concentrated in vacuo to a solid. These solids were slurried with
ethanol, filtered and the remaining mass was washed with additional
portions (2.times.) of ethanol. The combined filtrates were
concentrated in vacuo to afford the title compound (I-1 h), which
was taken on the next step without further purification.
Preparation of Intermediate 2,2-Difluoro-propionic acid ethyl ester
(I-3a)
[0184] ##STR31##
[0185] (Diethylamino)sulfur trifluoride (125 g, 780 mmol) was added
dropwise to stirred, cooled (0.degree. C.) ethyl pyruvate (71 ml,
650 mmol). The reaction was allowed to warm to ambient temperature
overnight, then quenched by slowly pouring over an ice/water
mixture and extracted with diethyl ether. The organic phase was
washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in
vacuo. The resulting oil was fractionally distilled at ambient
pressure (110-115.degree. C.) to afford the title compound (I-3a)
as a colorless oil, 55.6 gm.
Preparation of Intermediate
2,2-Difluoro-N-(2-hydroxy-ethyl)-propionamide (I-3b)
[0186] ##STR32##
[0187] 2,2-Difluoro-propionic acid ethyl ester I-3a (10.1 g, 70
mmol) was added dropwise to stirred, cooled (0.degree. C.)
ethanolamine (4.4 ml, 70 mmol) and the resulting solution was
allowed to stir at ambient temperature for 4 hours. Concentration
of the reaction mixture in vacuo afforded the title compound (I-3b)
as a solid, 11.2 g.
Preparation of Intermediate 2-(2,2-Difluoro-propylamino)-ethanol
(I-3c)
[0188] ##STR33##
[0189] To a stirred solution of lithium aluminum hydride (5.5 g,
146 mmol) in diethyl ether (85 ml) was added a solution of
2,2-difluoro-N-(2-hydroxy-ethyl)-propionamide (11.2 g, 73 mmol) in
diethyl ether (55 ml) dropwise at such a rate to maintain a gentle
reflux. After an additional 1.5 hours, the reaction was quenched
with sodium sulfate decahydrate, diluted with ethyl acetate and
allowed to stir for 18 hours. The mixture was filtered with the aid
of diatomaceous earth, washing with ethyl acetate. The filtrate was
concentrated in vacuo and fractionally distilled (11 torr,
collecting fractions distilling at 75-88.degree. C.) to afford the
title compound (I-3c) as a colorless oil, 4.5 g.
Preparation of Intermediate
1-(2-Chloro-phenyl)-5-(4-chloro-phenyl)-4-hydroxy-1H-pyrazole-3-carboxyli-
c acid (2,2-difluoro-propyl)-(2-hydroxy-ethyl)-amide (I-3d)
[0190] ##STR34##
[0191] A stirred mixture of
1-(2-chloro-phenyl)-5-(4-chloro-phenyl)-4-hydroxy-1H-pyrazole-3-carboxyli-
c acid ethyl ester I-3c (15 g, 40 mmol) and
2-(2,2-difluoro-propylamino)-ethanol (16.5 g, 120 mmol) were heated
at 125.degree. C. for 18 hours. The reaction solution was cooled,
diluted into ethyl acetate, washed with 1N aq. HCl, brine, dried
(Na.sub.2SO.sub.4) and concentrated in vacuo. The resulting oil was
chromatographed on silica gel (20% to 40% ethyl acetate/hexanes) to
afford the title compound (I-3d) as an oil, 10 g.
Preparation of Intermediate
4-Amino-1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-Pyrazole-3-carboxylic
Acid Ethyl Ester (I-4a)
[0192] ##STR35##
[0193] To a solution of LiN(TMS).sub.2 (1.0 M in THF, 100 ml, 100
mmol) in 400 ml diethyl ether at -78.degree. C. under nitrogen,
1-(4-chlorophenyl)ethanone (14.3 ml, 110 mmol) in 80 ml ether was
added dropwise via addition funnel. After the addition was
complete, the reaction mixture was stirred at -78.degree. C. for 40
minutes. Oxalic acid diethyl ester (14.3 ml, 105 mmol) was added in
one portion via syringe. The reaction mixture was warmed to room
temperature and stirred overnight. The pale white precipitate that
formed was collected by filtration. The solid was dried in vacuo to
give 4-(4-chlorophenyl)-2-hydroxy-4-oxobut-2-enoic acid ethyl ester
lithium salt (24.0 g, 92%).
[0194] A portion of the product from the previous step (10 g, 38.37
mmol) was dissolved in 400 ml acetic acid. After the solution was
cooled to 10.degree. C. with an ice-water bath, a concentrated
aqueous solution of sodium nitrite (2.86 g, 40.29 mmol) was added
dropwise, keeping the temperature between 10.degree. and 15.degree.
C. The reaction mixture was stirred for another 45 minutes, and
2-chlorophenylhydrazine HCl salt (8.5 g, 46.04 mmol) was added in
portions. Stirring was continued for 3 hours. Upon completion of
the reaction, the reaction mixture was poured into 600 ml ice-cold
water. A yellow solid precipitated and after 2 hours it was
collected, washed with water and dried to give crude
4-(4-chlorophenyl)-2-[(2-chlorophenyl)hydrazono]-3-nitroso-4-oxobutyric
acid ethyl ester which was used in the next step without further
purification.
[0195] The yellow solid obtained from last step was redissolved
into i-PrOH and 1 ml concentrated H.sub.2SO.sub.4 was added. The
reaction mixture was heated to 60.degree. C. for 3 hours. After
cooling to room temperature, the reaction mixture was poured into
ice-NaHCO.sub.3 (saturated aqueous). The precipitate was collected
by filtration and dried to give
1-(2-chlorophenyl)-5-(4-chlorophenyl)-4-nitroso-1H-pyrazole-3-carboxylic
acid ethyl ester. It was used for next step without further
purification.
[0196] The product obtained from the last step was dissolved in 200
ml of ethyl acetate and 200 ml of water. Sodium dithionite was
added until the disappearance of
1-(2-chlorophenyl)-5-(4-chlorophenyl)-4-nitroso-1H-pyrazole-3-carboxylic
acid ethyl ester was confirmed by TLC (ethyl acetate/hexane,
50/50). Then the organic layer was separated and the aqueous layer
was extracted with ethyl acetate. The combined organic layers were
dried over magnesium sulfate and the solvent was removed in vacuo.
The red solid obtained was further purified by plug filtration
(silica, ethyl acetate/hexane, 50/50) to give
4-amino-1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carbo-
xylic acid ethyl ester I-4a (21.86 g, 76%). MS: 376.1
(M+1).sup.+.
Preparation of Intermediate
4-(2-tert-Butoxycarbonylaminoethylamino)-1-(2-chlorophenyl)-5-(4-chloroph-
enyl)-1H-pyrazole-3-carboxylic Acid Ethyl Ester (I-4b)
[0197] ##STR36##
[0198] To a solution of
4-amino-1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylic
acid ethyl ester (I-4a, 1.88 g, 5 mmol) and (2-oxo-ethyl)-carbamic
acid tert-butyl ester (1590 mg, 10 mmol) in 1,2-dichloroethane (60
ml) at room temperature was added glacial acetic acid (858
microliters, 15 mmol) and NaBH(OAc).sub.3 (2540 mg, 12 mmol). The
reaction mixture was stirred for 23 hours, then additional
(2-oxo-ethyl)-carbamic acid tert-butyl ester (500 mg, 3.14 mmol)
and NaBH(OAc).sub.3 (1000 mg, 4.7 mmol) were added. The reaction
mixture was stirred for an additional 24 hours, quenched with 1 N
NaOH, and diluted with CH.sub.2Cl.sub.2. The organic layer was
separated and the aqueous layer extracted with CH.sub.2Cl.sub.2.
The combined organic phases were washed with 0.5 M aqueous citric
acid, 1 N NaOH, and saturated aqueous NaCl, dried, and concentrated
in vacuo. The crude residue was purified on a Biotage 40+M column
using CH.sub.2Cl.sub.2/MeOH (100:1) to give the desired product
I-4b (900 mg): +ES MS (M+1) 519.5.
Preparation of Intermediate
4-(2-tert-Butoxycarbonylaminoethylamino)-1-(2-chlorophenyl)-5-(4-chloroph-
enyl)-1H-pyrazole-3-carboxylic Acid (I-4c)
[0199] ##STR37##
[0200] To a solution of
4-(2-tert-butoxycarbonylaminoethylamino)-1-(2-chlorophenyl)-5-(4-chloroph-
enyl)-1H-pyrazole-3-carboxylic acid ethyl ester (I-4b, 900 mg) in
ethanol (50 ml) at room temperature was added 1 N KOH (25 ml). The
reaction mixture was heated at 50.degree. C. for 2 hours, cooled to
room temperature, diluted with saturated aqueous NaCl, and
acidified to pH 3 with 3N HCl. The aqueous solution was extracted
with EtOAc (2.times.) and the combined organic extracts were dried
and concentrated in vacuo to give the product I-4c as an amorphous
glass (879 mg): +ES MS (M+1) 491.5.
Preparation of Intermediate
4-(2-Aminoethylamino)-1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-
-carboxylic Acid (I-4d)
[0201] ##STR38##
[0202] A solution of
4-(2-tert-butoxycarbonylaminoethylamino)-1-(2-chlorophenyl)-5-(4-chloroph-
enyl)-1H-pyrazole-3-carboxylic acid (I-4c, 879 mg) in 2:1 conc
HCl/EtOH (12 ml) was allowed to stand at room temperature for 17
hours. The reaction mixture was concentrated under vacuum to give
the desired product I-4d: +ES MS (M+1) 391.4.
Preparation of Intermediate
3-(4-Chlorophenyl)-2-(2-chlorophenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3--
e][1,4]diazepin-8-on (I-4e)
[0203] ##STR39##
[0204] Diisopropylethylamine (209 microliters, 1.2 mmol) was added
slowly via syringe to a solution of
4-(2-aminoethylamino)-1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-
-carboxylic acid (I-4d, 228 mg, 0.53 mmol) and HATU (456 mg, 1.2
mmol) in anhydrous DMF (30 ml) at room temperature. The reaction
mixture was stirred for 17 hours, diluted with saturated aqueous
NaCl, and extracted with EtOAc (2.times.). The combined EtOAc
extracts were washed with 0.5 M citric acid, 1 M K.sub.2CO.sub.3,
and saturated aqueous NaCl, dried and concentrated in vacuo. The
residue was slurried with CH.sub.2Cl.sub.2 and the organic solvent
decanted, a process that was repeated twice. The combined organic
solutions were concentrated in vacuo and the residue was purified
on a chromatotron using 4 mm plates and a solvent gradient of 100%
EtOAc to 10:1 EtOAC/MeOH to give I-4e as a colorless solid (82 mg):
+ES MS (M+1) 373.4.
Preparation of
3-(4-Chlorophenyl)-2-(2-chlorophenyl)-4-trifluoroacetyl-4,5,6,7-tetrahydr-
o-2H-pyrazolo[4,3-e][1,4]diazepin-8-one (I-4f)
[0205] ##STR40##
[0206] To a solution of
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3--
e][1,4]diazepin-8-on (I-4e, 49 mg, 0.13 mmol) and NEt.sub.3 (21
microliters, 0.15 mmol) in 1,2-dichloroethane (1.5 ml) at 0.degree.
C. was slowly added triflic anhydride (20 microliters, 0.144 mmol).
The reaction mixture was allowed to warm to room temperature and
stir for 1 hour. Removal of an aliquot for analysis via APCI mass
spectrometry revealed only starting material present. Additional
NEt.sub.3 (31 mg) and triflic anhydride (50 mg) were added. The
reaction mixture was stirred at room temperature for 1 hour and
then was diluted with saturated aqueous NaCl. The organic layer was
separated and the aqueous layer was extracted twice with
CH.sub.2Cl.sub.2. The combined organic extracts were washed with
0.5 M citric acid, 1 M K.sub.2CO.sub.3, and saturated aqueous NaCl,
dried, and concentrated in vacuo. The crude residue was purified
via a chromatotron using 1 mm plates and 100% EtOAc as solvent to
give I-4f (45 mg): +ES MS (M+1) 469.2.
Preparation of
(L)-4-[(1-tert-Butoxycarbonyl-pyrrolidin-2-ylmethyl)-amino]-1-(2-chloroph-
enyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylic Acid Ethyl Ester
(I-5a)
[0207] ##STR41##
[0208] Glacial acetic acid (57 microliters, 1 mmol) and
NaBH(OAc).sub.3 (318 mg, 1.5 mmol) were added to a solution of
4-amino-1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylic
acid ethyl ester (376 mg, 1 mmol) and
(L)-2-formyl-pyrrolidine-1-carboxylic acid tert-butyl ester
(N-Boc-L-prolinal, 239 mg, 1.2 mmol) at room temperature in
1,2-dichloroethane (10 ml). The reaction mixture was stirred for 17
hours and quenched with 1 M NaOH (0.5 ml). The organic layer was
separated and the aqueous layer extracted once with
CH.sub.2Cl.sub.2. The combined organic solution was washed with
saturated aqueous NaCl, dried, and concentrated in vacuo. The
residue was purified on a chromatotron using 4 mm plates and a
solvent gradient of 100% CH.sub.2Cl.sub.2 to 20:1
CH.sub.2Cl.sub.2/MeOH to give the desired product I-5a as a
colorless white oil (207 mg): +ES MS (M+1) 559.5.
Preparation of
1-(2-Chlorophenyl)-5-(4-chlorophenyl)-4-[(pyrrolidin-2-ylmethyl)-amino]-1-
H-pyrazole-3-carboxylic Acid (I-5b)
[0209] ##STR42##
[0210] A solution of
(L)-4-[(1-tert-butoxycarbonyl-pyrrolidin-2-ylmethyl)-amino]-1-(2-chloroph-
enyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylic acid ethyl ester
(I-5a, 207 mg, 0.37 mmol) in 2:1 EtOH/1 N KOH (6 ml) was stirred at
50.degree. C. for 2 hours. After cooling to room temperature, the
reaction mixture was acidified with conc. HCl (.about.2 ml),
stirred for 4 hours, and concentrated under vacuum to give I-5b as
a white solid: +ES MS (M+1) 431.4.
Preparation of 4-(4-chlorophenyl)-2-hydroxy-4-oxo-but-2-enoic acid
ethyl ester lithium salt (I-6a)
[0211] ##STR43##
[0212] To tert-butyl methyl ether (350 ml) was added 149 ml of
lithium bis(trimethylsilyl)-amide (1.0 M in tetrahydrofuran, 149
mmol) at room temperature. The resulting solution was cooled to
-75.degree. C. 1-(4-chlorophenyl)ethanone (23.28 g, 150.6 mmoles)
was added as a solution in 23 ml of tert-butyl methyl ether over 3
minutes, keeping internal temperature less than -70.degree. C. The
reaction solution was held for 1 hour at -75.degree. C., then
diethyl oxalate (22.0 g, 150 mmol) was added neat over 5 minutes
keeping internal temperature less than -70.degree. C. The clear
dark orange reaction solution was warmed to room temperature over 4
hours. Note: Product began to precipitate at -3.degree. C. The
reaction was allowed to stir for 15 hours at room temperature,
followed by isolation of precipitated product by filtration. The
filtercake was washed with 100 ml of room temperature tert-butyl
methyl ether and then dried at 60.degree. C. and 10 mm for 1 hour
to give 4-(4-chlorophenyl)-2-hydroxy-4-oxo-but-2-enoic acid ethyl
ester lithium salt I-6a (36.72 g, 94%) as a powdery yellow
solid.
[0213] .sup.1H-NMR (DMSO-d.sub.6) .delta. 7.80 (d, 1.94H, J=8.7
Hz), 7.66 (d, 0.06H, J=8.7 Hz), 7.43 (d, 1.94H, J=8.7 Hz), 7.31 (d,
0.06H, J=8.3 Hz), 6.37 (s, 0.97H), 5.22 (s, 0.03H), 4.10 (q, 1.94H,
J=7.05 Hz), 4.00 (q, 0.06H, J=7.05 Hz), 1.20 (t, 2.91H, J=7.05 Hz),
1.15 (t, 0.09H, J=7.05 Hz). Shows a 97:3 mixture of geometric
isomers.
[0214] Mass Spec (ESI): M+1=255.2 (mass of neutral compound)
Preparation of
1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylic acid
(I-6b)
[0215] ##STR44##
[0216] 4-(4-Chlorophenyl)-2-hydroxy-4-oxo-but-2-enoic acid ethyl
ester lithium salt I-6a (30.26 g, 116 mmoles) was suspended in 242
ml of ethanol. 2-Chlorophenylhydrazine hydrochloride (20.88 g, 116
mmoles) was added portionwise as a solid over 45 minutes while
maintaining an internal temperature between 30-40.degree. C. Note:
Reaction mixture goes from a yellow suspension to a dark orange
suspension. Reaction stirred for 3 hours while maintaining internal
temperature between 25-35.degree. C. An aqueous potassium hydroxide
solution (148 ml of 1.8 M solution, 266 mmoles) was added over 20
minutes while maintaining an internal temperature between
20-30.degree. C. The reaction mixture was held for 2.5 hours. Note:
Within 30 minutes of potassium hydroxide solution addition,
reaction turned almost clear, very dark rust orange in color.
Aqueous hydrochloric acid (85 ml of 3.9 M solution, 331 mmoles) was
added over 15 minutes while maintaining reaction temperature
between 20-30.degree. C. Note: Product precipitated during
hydrochloric acid addition. The precipitated product was granulated
for 16 hours at room temperature. The crude product was isolated by
filtration and the filtercake was washed with 150 ml of water.
Note: Filtercake was a yellowish orange solid. After air-drying for
30 minutes, the filtercake was suspended in 480 ml of methanol.
This suspension was heated to reflux to give a clear dark orange
solution (all solids in solution within 1 hour of reaching reflux)
that was held at reflux for 8 hours. The solution was cooled over 4
hours to room temperature, during which time product had
precipitated from solution. The reaction mixture was held at room
temperature for 10 hours, followed by cooling to 0.degree. C., and
stirring for 1.5 hours. Collection of the precipitate by
filtration, washing the resulting filtercake with 150 ml of
ice-chilled methanol, and drying at 60.degree. C. and 1 mm for 3
hours afforded
1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylic acid
I-6b (29.28 g, 76%) as an off-white solid.
[0217] .sup.1H-NMR (CD.sub.3CN): .delta. 7.58-7.45 (m, 4H), 7.31
(d, 2H, J=8.7 Hz), 7.21 (d, 2H, J=8.7 Hz), 7.10 (s, 1H). Mass Spec
(ESI): M+1=333.2
Preparation of
1-(2-chlorophenyl)-5-(4-chlorophenyl)-4-hydroxy-1H-pyrazole-3-carboxylic
acid (I-6c)
[0218] ##STR45##
[0219]
1-(2-Chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylic acid
I-6b (628.1 g, 1.88 mol) was dissolved in tetrahydrofuran (11
liters) to give a clear light orange solution. This solution was
cooled to -78.degree. C. followed by the addition of hexyllithium
(2.0 M solution in hexanes, 2.07 liters, 4.14 mol) over a period of
2 hours while keeping internal temperature less than -70.degree. C.
Note: During addition of the first equivalent of hexyllithium,
reaction solution remained clear orange, then during addition of
second equivalent of hexyllithium, reaction solution turned brown
and then very dark green. Reaction mixture held for 20 minutes at
-74.degree. C., then warmed to -50.degree. C. over 30 minutes and
held for 1 hour at this temperature. The reaction was cooled back
to less than -70.degree. C., followed by the addition of neat
trimethylborate (238 g, 2.01 moles) over 3 minutes while keeping
temperature less than -68.degree. C. The reaction solution was then
warmed to room temperature over 3 hours. Note: Reaction remained
very dark green until reaching room temperature where it turned
clear dark orange. Aqueous sodium hydroxide (750 mL of 3.0 M, 2.25
mol) was added over 5 minutes to crude reaction solution while
maintaining an internal temperature of 10-15.degree. C.
Concentrated aqueous hydrogen peroxide (253 g, 30 wt %, 2.01 moles)
was then added over a period of 30 minutes while maintaining an
internal temperature between 10-20.degree. C. The reaction was
allowed to warm to room temperature and stirred for 3.5 hours.
Water (3 liters) was added followed by addition of concentrated
aqueous hydrochloric acid (545 ml, 12.1 M, 6.59 mol) over 15
minutes while maintaining a temperature of 20-30.degree. C. Note:
pH of crude reaction solution is .about.2.5. The tetrahydrofuran
and aqueous layers were separated and the aqueous layer was
extracted with 4 liters of tert-butyl methyl ether. The
tetrahydrofuran and tert-butyl methyl ether layers were combined,
washed with 4 liters of brine, and dried over 2.5 Kg of
Na.sub.2SO.sub.4. The crude solution was concentrated in vacuo to a
thick orange oil containing some fine solids. The crude orange oil
was then added to 5 liters of methanol, causing a bright yellow
precipitate to crystallize from solution. The precipitated product
was granulated for 20 hours at room temperature followed by cooling
to 0.degree. C. and stirring for 1 hour. The crude product was
isolated by filtration and the resulting filtercake was washed with
1 liter of ice-chilled methanol. The filtercake was air-dried for
18 hours. This crude product (390 g) was suspended in 2.1 liters of
2-propanol followed by heating to reflux to give a clear
yellow/orange solution. Solution held at reflux for 1 hour, then
cooled over a period of 5 hours to 3.degree. C. and stirred for 1
hour. The recrystallized product was isolated by filtration and the
resulting filtercake was washed with 900 ml of ice-chilled
2-propanol, followed by air-drying for 18 hours. The product was
oven-dried for 18 hours at 60.degree. C. and 10 mm to afford
1-(2-chlorophenyl)-5-(4-chlorophenyl)-4-hydroxy-1H-pyrazole-3-carboxylic
acid I-6c (282.9 g, 43%) as an off-white solid.
[0220] .sup.1H-NMR (CD.sub.3CN): .delta. 7.55-7.44 (m, 4H), 7.31
(d, 2H, J=8.7 Hz), 7.20 (d, 2H, J=8.7 Hz). Mass Spec (ESI):
M+1=349.2
Preparation of
4-Acetoxy-1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylic
acid (I-6d)
[0221] ##STR46##
[0222]
1-(2-Chlorophenyl)-5-(4-chlorophenyl)-4-hydroxy-1H-pyrazole-3-carb-
oxylic acid 1-6c (572.0 g, 1.64 mol) was combined with 8 liters of
methylene chloride to give an off-white suspension.
N,N-Diisopropylethylamine (427.9 g, 3.29 mol) was added over 15
minutes while keeping the temperature between 20-25.degree. C. A
clear yellow solution resulted. Acetic anhydride (334.5 g, 3.24
mol) was added over 5 minutes keeping the temperature between
20-25.degree. C. The reaction was stirred at room temperature for
16 hours. The crude reaction solution was washed twice with 4
liters portions of 0.5 M citric acid and once with 4 liters of
brine. The crude solution was concentrated in vacuo to a total
volume of 1 liter. This milky suspension was then added to 4 liters
of hexanes causing the desired product to precipitate instantly.
The solids were granulated for 30 minutes and then collected by
filtration. The filtercake was rinsed with 3 liters of hexanes and
then air-dried for 16 hours. The isolated product was then further
dried at 60.degree. C. and 8 mm for 2 hours.
4-Acetoxy-1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylic
acid 1-6d (601.6 g, 94%) was isolated as a powdery, off-white
solid.
[0223] .sup.1H-NMR (CD.sub.2Cl.sub.2): .delta. 7.50 (d, 1H, J=7.0
Hz), 7.47-7.37 (m, 3H), 7.28 (d, 2H, J=8.7 Hz), 7.12 (d, 2H, J=8.7
Hz), 2.26 (s, 3H). Mass Spec (ESI): M+1=391.2
Preparation of Acetic acid
1-(2-chlorophenyl)-5-(4-chlorophenyl)-3-[(2,2-difluoro-propyl)-(2-hydroxy-
ethyl)-carbamoyl]-1H-pyrazol-4-yl ester (I-6e)
[0224] ##STR47##
[0225]
4-Acetoxy-1-(2-chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carb-
oxylic acid I-6d (581.0 g, 1.48 mol) was dissolved in 10 liters of
methylene chloride to give a pale yellow, slightly opaque solution.
The solution was filtered through Celite.RTM. to give a clear green
colored solution. 2-Chloro-4,6-dimethoxy-1,3,5-triazine (296.8 g,
1.64 mol) was added in one portion as a solid at room temperature
to give an opaque suspension (addition is slightly endothermic).
4-Methylmorpholine (182.9 g, 1.80 mol) was added over 15 minutes
while keeping the temperature between 18-22.degree. C. (reaction
returned to being yellow in color). Reaction stirred for 3 hours at
room temperature, then 2-(2,2-difluoropropylamino)-ethanol (228.3
g, 1.64 mol) was added neat over 10 minutes while keeping the
temperature between 20-25.degree. C. The reaction was stirred for
15 hours, then washed twice with 6 liter portions of 10% citric
acid and once with 5 liters of brine. The crude product solution
was concentrated in vacuo to a thick orange oil, then reconstituted
in 4 liters of isopropyl ether. After removing 1 liter of
distillates precipitate began to form. To the crude product
suspension was added 1.5 liters of isopropyl ether and then the
mixture was stirred at room temperature for 1 hour. The
precipitated solids were collected by filtration and the resulting
filtercake was rinsed with 2 liters of room temperature isopropyl
ether, followed by air-drying for 16 hours. Acetic acid
1-(2-chlorophenyl)-5-(4-chlorophenyl)-3-[(2,2-difluoro-propyl)-(2-hy-
droxyethyl)-carbamoyl]-1H-pyrazol-4-yl ester I-6e (603.0 g, 78%)
was isolated as a granular off-white solid.
[0226] .sup.1H-NMR (CD.sub.2Cl.sub.2): .delta. 7.50-7.31 (m, 4H),
7.28 (d, 2H, J=8.3 Hz), 7.14 (d, 2H, J=8.7 Hz), 4.41-3.41 (m,
various rotamers, 7H), 2.21 (s, 3H), 1.65 (t, 3H, J.sub.HF=19.5
Hz).
[0227] Mass Spec (ESI): M+1=512.2
Preparation of Acetic acid
3-[(2-chloroethyl)-(2,2-difluoropropyl)-carbamoyl]-1-(2-chlorophenyl)-5-(-
4-chlorophenyl)-1H-pyrazol-4-yl-ester (I-6f)
[0228] ##STR48##
[0229] Method A: Acetic acid
1-(2-chlorophenyl)-5-(4-chlorophenyl)-3-[(2,2-difluoro-propyl)-(2-hydroxy-
-ethyl)-carbamoyl]-1H-pyrazol-4-yl ester I-6e (580.6 g, 1.12 mol)
was dissolved in 10 liters of methylene chloride to give a clear
pale yellow solution. After cooling to 0.degree. C.,
methanesulfonyl chloride (142.4 g, 1.21 mol) was added neat over 5
minutes, followed by addition of neat N,N-diisopropylethylamine
(167.9 g, 1.29 mol) over 25 minutes, while keeping the temperature
less than 5.degree. C. After stirring 20 minutes at <5.degree.
C., the reaction was warmed to room temperature and stirred for 14
hours. The crude reaction solution was washed twice with 4.5 liter
portions of 10% citric acid and once with 4 liters of brine. The
crude product solution was concentrated in vacuo to give a crude
solid, then 2 liters of methanol was added followed by stirring for
1 hour. About half of the crude solid had dissolved in and then
crystallized from the methanol. This material was collected by
filtration and the resulting filtercake was rinsed with 300 ml of
room temperature methanol. This first crop of material was dried at
50.degree. C. and 10 mm for 2 hours to give 245.2 g, 41.2% of the
title compound as an off-white solid. The crude solid that had not
dissolved in and crystallized from methanol was redissolved in 1
liter of methylene chloride, then concentrated to a viscous
brownish oil. The methanol mother liquor left over from the first
crop was concentrated to a total volume of 800 ml and was then
combined with the viscous brownish oil. This mixture was warmed in
a 40.degree. C. waterbath until a clear solution was obtained, then
the resulting solution was cooled to 0.degree. C. and stirred for
30 minutes, resulting in product precipitation. The precipitate was
collected by filtration, and the resulting filtercake was washed
with 200 ml of ice-chilled methanol, followed by air-drying for 16
hours. The second crop material (290.9 g, 48.8%) was isolated as an
off-white solid. The overall combined yield of first and second
crops of acetic acid
3-[(2-chloroethyl)-(2,2-difluoropropyl)-carbamoyl]-1-(2-chlorophenyl)-5-(-
4-chlorophenyl)-1H-pyrazol-4-yl-ester I-6f was 536.1 g (90%).
[0230] Method B:
1-(2-Chlorophenyl)-5-(4-chlorophenyl)-1H-pyrazole-3-carboxylic acid
I-6b (90.8 g, 260 mmol) was dissolved in 1.7 liters of methylene
chloride, giving an off-white suspension. 4-Methylmorpholine (58.5
g, 576 mmol) was added, giving a clear yellow solution, followed by
addition of acetyl chloride (22.6 g, 284 mmol) over 10 minutes
while maintaining a temperature between 20-30.degree. C. The
reaction was stirred for 7 hours at room temperature, then cooled
to 0.degree. C. 2-(2,2-Difluoropropylamino)-ethanol (39.8 g, 286
mmol) was added neat over 1 minute followed by addition of
2-Chloro-4,6-dimethoxy-1,3,5-triazine (49.0 g, 271 mmol)
portionwise as a solid over 1 minute. The reaction was allowed to
slowly warm to room temperature over a period of 5 hours, followed
by stirring for 12 hours at room temperature. The crude reaction
solution was washed twice with 900 ml portions of 0.5 M citric acid
and once with 900 ml of brine. Residual water was azeotropically
removed through two cycles concentrating off methylene chloride and
then adding more methylene chloride. The final crude methylene
chloride solution volume was 1.2 liters. This solution was cooled
to -2.degree. C. followed by addition of neat methanesulfonyl
chloride (36.0 g, 311 mmol) and then addition of neat
N,N-diisopropylethylamine (42.1 g, 324 mmol) over a 10 minute
period while maintaining a reaction temperature less than
10.degree. C. The reaction solution was warmed to room temperature
over 1 hour, followed by stirring for 20 hours, then washing the
methylene chloride solution twice with 800 ml portions of 0.5 M
citric acid and once with 800 ml brine. Product rich methylene
chloride layer was clear dark orange in appearance (.about.1.3
liters total volume). Crude solution was concentrated in vacuo to
.about.300 ml, followed by addition of 1 liter of methanol.
Resulting solution was concentrated in vacuo in a 30.degree. C.
waterbath by removing 900 ml of distillates. Another 800 ml portion
of methanol was added followed by a final concentration in vacuo
(30.degree. C. waterbath) to remove 700 ml of distillates. The
final total volume was .about.500 ml. The product rich concentrated
solution was held at room temperature for 1 hour (solution was
initially hazy, dark orange in appearance, then solids precipitated
after .about.15 minutes). The mixture was cooled to -10.degree. C.
and stirred for 1 hour while maintaining temperature less than
0.degree. C. The precipitated solids were collected by filtration,
and the resulting filtercake was washed with 50 ml of ice-chilled
methanol, followed by air-drying for 15 hours. The isolated solids
were further dried at 60.degree. C. and 1 mm for 2 hours (loss on
drying was only 0.4 g) to give acetic acid
3-[(2-chloroethyl)-(2,2-difluoropropyl)-carbamoyl]-1-(2-chlorophenyl)-5-(-
4-chlorophenyl)-1H-pyrazol-4-yl-ester I-6f (104.0 g, 75%) as a
white solid.
[0231] .sup.1H-NMR (CD.sub.2Cl.sub.2): .delta. 7.49-7.47 (m, 1H),
7.44-7.40 (m, 1H), 7.37-7.33 (m, 2H), 7.28 (d, 2H, rotamers, J=8.7
Hz), 7.14 (d, 2H, rotamers, J=8.7 Hz), 4.46 (t, 0.72H, J cannot be
determined), 4.14 (t, 1.28H, J cannot be determined), 3.97 (t,
1.28H, J.sub.HF=13.0 Hz), 3.87 (t, 0.72H, J=6.4 Hz), 2.22 (s,
1.08H), 2.20 (s, 1.92H), 1.62 (t, 3H, rotamers, J.sub.HF=19.5 Hz).
Two major rotamers present in a .about.1.7:1 ratio.
[0232] Mass Spec (ESI): M+1=530.2
Preparation of
1-(2-chlorophenyl)-5-(4-chlorophenyl)-4-hydroxy-1H-pyrazole-3-carboxylic
acid (2-chloroethyl)-(2,2-difluoropropyl)-amide (I-6g)
[0233] ##STR49##
[0234] Acetic acid
3-[(2-chloroethyl)-(2,2-difluoropropyl)-carbamoyl]-1-(2-chlorophenyl)-5-(-
4-chlorophenyl)-1-H-pyrazol-4-yl-ester I-6f (3.55 g, 6.69 mmol) was
dissolved in 90 ml of methanol with warming in a 40.degree. C.
waterbath to give a clear colorless solution. The resulting
solution was cooled to 0.degree. C. (still a clear, colorless
solution), followed by addition of K.sub.2CO.sub.3 (1.02 g, 7.31
mmol) in one portion as a solid (reaction mixture goes from
colorless to yellow). Reaction stirred for 30 minutes at 0.degree.
C., followed by addition of concentrated hydrochloric acid (1.2 ml
of 12.1 M, 14.5 mmol). Note: upon neutralization, reaction turns
colorless and clear, then product begins to precipitate. The
reaction was warmed to room temperature, then 45 ml of water was
added, followed by stirring for 2.5 hours. The precipitated solids
were collected by filtration and the resulting filtercake was
washed with 50 ml of room temperature 2:1, methanol:water.
Collected solids were dried at 50.degree. C. and 10 mm for 1 hour
to give
1-(2-chlorophenyl)-5-(4-chlorophenyl)-4-hydroxy-1H-pyrazole-3-carboxylic
acid (2-chloroethyl)-(2,2-difluoropropyl)-amide I-6 (2.86 g, 87%)
as a white solid.
[0235] .sup.1H-NMR (CD.sub.2Cl.sub.2): .delta. 9.67 (s, 0.52H),
9.57 (s, 0.48H), 7.51-7.48 (m, 1H), 7.46-7.41 (m, 1H), 7.39-7.31
(m, 2H), 7.24 (d, 2H, rotamers, J=8.7 Hz), 7.17 (d, 2H, rotamers,
J=8.7 Hz), 4.75 (t, 0.52H, J.sub.HF=13 Hz), 4.47 (t, 0.48H, J=6
Hz), 4.08 (t, 0.48H, J.sub.HF=13 Hz), 3.94 (t, 0.52H, J=6 Hz),
3.84-3.79 (m, 2H), 1.66 (t, 1.44H, J.sub.HF=19.3 Hz), 1.59 (t,
1.56H, J.sub.HF=19.1 Hz). Two major rotamers present in a
.about.1.07:1 ratio.
[0236] Mass Spec (ESI): M+1=488.2
Example 1
Preparation of
3-(4-Chlorophenyl)-2-(2-chlorophenyl)-7-(2,2,2-trifluoroethyl)-6,7-dihydr-
o-2H,5H-4-oxa-1,2,7-triaza-azulen-8-one (1A-1)
[0237] ##STR50##
[0238] A solution of
5-(4-chlorophenyl)-1-(2-chlorophenyl)-4-[2-(2,2,2-trifluoroethylamino)-et-
hoxy]-1H-pyrazole-3-carboxylic acid, hydrochloride I-1h (45 mg, 0.1
mmol), triethylamine (0.05 ml) and 1-propanephosphoric acid cyclic
anhydride (0.1 ml, 0.14 mmol) in 1,2-dichloroethane (0.6 ml) were
stirred for 8 hours. The reaction was diluted into ethyl ether,
washed 1 N aqueous hydrochloric acid, saturated aqueous sodium
bicarbonate, brine, dried (Na.sub.2SO.sub.4) and concentrated in
vacuo to afford the title compound (1A-1) as a white solid, 35 mg.
.sup.1H NMR in CDCl.sub.3 (ppm): .delta.7.52 (d, 1H), 7.38-7.34 (m,
2H), 7.23 (d, 2H), 7.15 (d, 2H), 4.47 (br s, 2H), 4.29 (br s, 2H),
3.91 (br s, 2H); ms (LCMS) m/z=456.3 (M+1).
[0239] The compounds listed below were prepared using procedures
analogous to those described above for the synthesis of Compound
1A-1 and outlined in Scheme I above using the appropriate starting
materials which are available commercially or prepared using
preparations well-known to those skilled in the art. ##STR51##
3-(4-Chlorophenyl)-2-(2-chlorophenyl)-7-isopropyl-6,7-dihydro-2H,5H-4-oxa-
-1,2,7-triaza-azulen-8-one (1A-2): m/z=416.4 (M+1) ##STR52##
3-(4-Chlorophenyl)-7-isopropyl-2-o-tolyl-6,7-dihydro-2H,5H-4-oxa-1,2,7-tr-
iaza-azulen-8-one (1A-3): m/z=396.4 (M+1) ##STR53##
3-(4-Chloro-phenyl)-2-o-tolyl-7-(2,2,2-trifluoro-ethyl)-6,7-dihydro-2H,5H-
-4-oxa-1,2,7-triaza-azulen-8-one (1A-4): m/z=436.2 (M+1)
Example 2
Preparation of
3-(4-Chloro-Phenyl)-2-(2-chloro-phenyl)-7-(2,2-difluoro-Propyl)-6,7-dihyd-
ro-2H,5H-4-oxa-1,2,7-triaza-azulen-8-one (2A-1)
[0240] ##STR54##
[0241] To a stirred solution of
1-(2-chloro-phenyl)-5-(4-chloro-phenyl)-4-hydroxy-1H-pyrazole-3-carboxyli-
c acid (2,2-difluoro-propyl)-(2-hydroxy-ethyl)-amide I-3d (10 g, 21
mmol), triphenyl phosphine (8.4 g, 31.5 mmol) in toluene (210 ml)
was added 1,1'-(azodicarbonyl)dipiperidine (8.0 g, 31.5 mmol).
After 18 hours, 20% ethyl acetate:hexanes (210 ml) was added, the
mixture was stirred at ambient temperature for 1 hour and filtered.
The filtrate was concentrated in vacuo and the resulting oil was
chromatographed on silica gel (20-70% ethyl acetate:hexanes) to
afford the title compound (2A-1) as a solid, 7.8 g. .sup.1H NMR in
CDCl.sub.3 (ppm) .delta. 7.53-7.50 (m, 1H), 7.38-7.33 (m, 3H),
7.24-7.21 (m, 2H), 7.16-7.13 (m, 2H), 4.45 (brs, 2H), 4.02 (t, 2H),
3.90 (br s, 2H), 1.69 (t, 3H); ms (LCMS) m/z=452.2 (M+1).
Combustion analysis calculated for: C, 55.77%; H, 3.79%; N, 9.29%.
Found: C, 55.69%; H, 3.52%; N, 9.13%.
[0242] The compounds listed below were prepared using procedures
analogous to those described above for the synthesis of Compound
2A-1 and outlined in Scheme III above using the appropriate
starting materials which are available commercially or prepared
using preparations well-known to those skilled in the art.
##STR55##
3-(4-Chloro-phenyl)-2-(2-chloro-phenyl)-7-(2,2-difluoro-butyl)-6,7-dihydr-
o-2H,5H-4-oxa-1,2,7-triaza-azulen-8-one (2A-2): m/z=466.1 (M+1)
##STR56##
3-(4-Chloro-phenyl)-7-(2,2-difluoro-butyl)-2-o-tolyl-6,7-dihydro-2H,5H-4--
oxa-1,2,7-triaza-azulen-8-one (2A-3): m/z=446.2 (M+1)
Example 3
Preparation of
3-(4-Chlorophenyl)-2-(2-chlorophenyl)-7-(2,2,2-trifluoroethyl)-4,5,6,7-te-
trahydro-2H-pyrazolo[4,3-e][1,4]diazepin-8-one (3A-1)
[0243] ##STR57##
[0244] A solution of
3-(4-chlorophenyl)-2-(2-chlorophenyl)-4-trifluoroacetyl-4,5,6,7-tetrahydr-
o-2H-pyrazolo[4,3-e][1,4]diazepin-8-one (I-4f, 47 mg, 0.10 mmol)
and NaH (5 mg, 60% dispersion, 0.12 mmol) in DMF (1.5 ml) was
stirred for 1 hour at room temperature under a N.sub.2 atmosphere,
then trifluoromethanesulfonic acid 2,2,2-trifluoroethyl ester (22
microliters, 0.15 mmol) was added dropwise. The reaction mixture
was stirred for 2 hours, quenched with saturated aqueous NaCl, and
extracted twice with EtOAc. The combined organic extracts were
washed with saturated aqueous NaHCO.sub.3 and saturated aqueous
NaCl, dried, and concentrated under vacuum. The crude residue was
purified on a chromatotron using 1 mm plates and a solvent gradient
of 1:1 hexanes/EtOAc to 100% EtOAc to 9:1 EtOAc/MeOH to give 3A-1
as an amorphous glass (6 mg): +ES MS (M+1) 455.4; .sup.1H NMR
(CD.sub.2Cl.sub.2) .delta. 7.50-7.37 (m, 4H), 7.32 (d, 2H, J=8.3
Hz), 7.18 (d, 2H, J=8.3 Hz), 4.29 (q, 2H, J=8.8 Hz), 3.84-3.79 (m,
2H), 3.60-3.55 (m, 2H).
Example 4
Preparation of
1-(4-Chlorophenyl)-2-(2-chlorophenyl)-5,6,7,7a,8,9-hexahydro-2H-2,3,4a,9--
tetraazacyclopenta[f]azulen-4-one (4A-1)
[0245] ##STR58##
[0246] Diisopropylethylamine (250 microliters, 2 mmol) was added to
a mixture of
1-(2-chlorophenyl)-5-(4-chlorophenyl)-4-[(pyrrolidin-2-ylmethyl)-amino]-1-
H-pyrazole-3-carboxylic acid (I-5b, 160 mg, 0.37 mmol) and HATU
(380 mg, 1 mmol) in DMF (50 ml) at room temperature. The reaction
mixture was stirred for 5 hours, quenched with saturated aqueous
NaCl, and extracted twice with EtOAc. The combined organic extracts
were washed with 0.5 M citric acid, 1 M K.sub.2CO.sub.3, and
saturated aqueous NaCl, dried, and concentrated in vacuo. The
residue was slurried in CH.sub.2Cl.sub.2 and the solvent decanted,
a process repeated once more with EtOAc. The combined organic
solutions were concentrated under vacuum and the residue was
purified on a chromatogram using 4 mm plates and a solvent gradient
of 10:1 EtOAc/MeOH to 5:1 EtOAc/MeOH to give 4A-1 as a white solid
(46 mg): +ES MS (M+1) 413.4; .sup.1H NMR (CD.sub.3OD) .delta. 7.58
(bs, 1H), 7.52-7.44 (m, 3H), 7.34 (d, 2H, J=7.6 Hz), 7.22 (d, 2H,
J=7.6 Hz), 3.93-3.82 (m, 2H), 3.64-3.50 (m, 2H), 3.10-2.99 (m, 1H),
2.38-2.29 (m, 1H), 2.04-1.96 (m, 1H), 1.95-1.74 (m, 2H).
[0247] The following demonstrates an alternative method for
preparing compounds of the present invention where A is nitrogen, B
is carbon and X is O.
Example 5
Preparation of
3-(4-Chlorophenyl)-2-(2-chlorophenyl)-7-(2,2-difluoropropyl)-6,7-dihydro--
2H,5H-4-oxa-1,2,7-triaza-azulen-8-one (I-5A)
[0248] ##STR59##
[0249] Acetic acid
3-[(2-chloroethyl)-(2,2-difluoropropyl)-carbamoyl]-1-(2-chlorophenyl)-5-(-
4-chlorophenyl)-1H-pyrazol-4-yl-ester I-6g (513.0 g, 0.97 mol) was
suspended in 9.7 liter of ethanol (gives a off-white suspension).
Cesium carbonate (348.0 g, 1.07 mol) was added portionwise as a
solid over 2 minutes while maintaining internal temperature between
21-27.degree. C. Note: Upon Cs.sub.2CO.sub.3 addition, reaction
mixture turned pale yellow (still a suspension). Reaction stirred
at room temperature for 19 hours, then the crude reaction mixture
was filtered through Celite.RTM. to remove insoluble solids, giving
a clear dark yellow filtrate. The Celite.RTM. filtercake was washed
with 2 liters of ethanol. The crude product solution was
concentrated in vacuo and gave a yellow solid. This solid was
reconstituted in 7 liters of methylene chloride and the resulting
mixture was washed once with 5 liters of half saturated aqueous
NH.sub.4Cl and once with 4 liters of brine. The product rich
methylene chloride layer was concentrated in vacuo to a total
volume of 2.5 liters. Note: The methylene chloride layer was clear
and dark resddish in color. The product rich methylene chloride
solution was treated with 105 g of Darco, followed by stirring at
reflux for 30 minutes. After cooling, the Darco was filtered off by
passing the solution through Celite.RTM.. Note: Crude product
solution is clear dark orange in appearance. The crude product
filtrate was concentrated in vacuo to a total volume of 1.1 liters.
This product rich methylene chloride solution was added over 20
minutes to 5 liters of cyclohexane while maintaining a reaction pot
temperature of 50-60.degree. C. Note: Halfway through methylene
chloride solution addition, precipitate came out of solution. After
complete addition, the methylene chloride solvent was
atmospherically removed (3.55 liters of distillates collected while
simultaneously adding 2 liters of cyclohexane to refluxing
solution) from the reaction mixture by heating to 79.degree. C.
(internal pot temperature) over a 2.5 hour period. Once internal
temperature reached the boiling point of cyclohexane, all of the
methylene chloride had been displaced. Note: The reaction mixture
took on a very dark pink/purple coloration with white solids
suspended. Reaction mixture held at 79.degree. C. for 10 minutes,
then cooled to 50.degree. C. and held for 13 hours, followed by
cooling to 30.degree. C. and holding for 4 hours. The precipitated
product was collected by filtration, and the resulting filtercake
was washed with 3 liters of room temperature cyclohexane, followed
by air-drying for 3.5 hours. The isolated solids were further dried
at 50.degree. C. and 2 mm for 15 hours (loss on drying was only 0.2
g) to give
3-(4-chlorophenyl)-2-(2-chlorophenyl)-7-(2,2-difluoropropyl)-6,7-dihydro--
2H,5H-4-oxa-1,2,7-triaza-azulen-8-one 1-5A (321.3 g, 73%) as an
off-white solid.
Recrystallization of
3-(4-Chlorophenyl)-2-(2-chlorophenyl)-7-(2,2-difluoropropyl)-6,7-dihydro--
2H,5H-4-oxa-1,2,7-triaza-azulen-8-one (I-5A)
[0250]
3-(4-Chlorophenyl)-2-(2-chlorophenyl)-7-(2,2-difluoropropyl)-6,7-d-
ihydro-2H,5H-4-oxa-1,2,7-triaza-azulen-8-one 1-5A (5.00 g, 11.1
mmol) was dissolved in 20 ml of methylene chloride to give a clear
orange solution. Darco KBB (0.5 g) was added followed by heating to
reflux and stirring for 1 hour. After cooling, the Darco KBB was
filtered off by passing the solution through Celite.RTM., giving a
clear light yellow filtrate. The Celite.RTM. filtercake was washed
with 10 ml of methylene chloride. The eluent was concentrated in
vacuo to give a total solution volume of .about.20 ml. The
concentrated methylene chloride solution was then diluted with 150
ml of 2-propanol to give a clear pale yellow solution. Methylene
chloride was removed from the resulting solution by atmospherically
distilling off 71 ml of distillates as solution was heated from
room temperature to 82.degree. C. (boiling point of 2-propanol).
The solution was then cooled over 3 hours from 82.degree. C. to
room temperature. Note: Solution became hazy around 34.degree. C.,
followed by precipitate formation. The mixture was stirred at room
temperature for 62 hours, then cooled to 0.degree. C. and stirred
for 2.5 hours before collecting the precipitate by filtration. The
resulting filtercake was washed with 80 ml of ice-chilled
2-propanol, then air-dried for 1 hour. The recrystallized
3-(4-Chlorophenyl)-2-(2-chlorophenyl)-7-(2,2-difluoropropyl)-6,7-dihydro--
2H,5H-4-oxa-1,2,7-triaza-azulen-8-one 1-5A (4.03 g, 81%) was
isolated as a pure white crystalline solid.
[0251] .sup.1H-NMR (CD.sub.2Cl.sub.2): .delta. 7.49-7.46 (m, 1H),
7.45-7.37 (m, 3H), 7.24 (d, 2H, rotamers, J=9.1 Hz), 7.16 (d, 2H,
rotamers, J=8.7 Hz), 4.44 (dd, 2H, J=5.2 Hz, 1.9 Hz), 3.98 (t, 2H,
J.sub.HF=13 Hz), 3.87 (t, 2H, J=3.7 Hz), 1.67 (t, 3H, J.sub.HF=19.1
Hz)
[0252] Mass Spec (ESI): M+1=452.2
[0253] In addition to the compounds described above in Examples
1-5, the following compounds may be prepared using the methods and
procedures generally described above in Scheme IV: [0254]
2-(2-Chlorophenyl)-3-(4-ethylphenyl)-7-(2,2,2-trifluoroethyl)-6,7-dihydro-
-2H,5H-4-oxa-1,2,7-triaza-azulen-8-one; [0255]
2-(2-Chlorophenyl)-7-(2,2-difluoropropyl)-3-(4-ethylphenyl)-6,7-dihydro-2-
H,5H-4-oxa-1,2,7-triaza-azulen-8-one; and [0256]
2-(2-Chlorophenyl)-3-(4-ethylphenyl)-7-isopropyl-6,7-dihydro-2H,5H-4-oxa--
1,2,7-triaza-azulen8-one
Pharmacological Testing
[0257] The utility of the compounds of the present invention in the
practice of the instant invention can be evidenced by activity in
at least one of the protocols described hereinbelow. The following
acronyms are used in the protocols described below.
[0258] BSA--bovine serum albumin
[0259] DMSO--dimethylsulfoxide
[0260] EDTA--ethylenediamine tetracetic acid
[0261] PBS--phosphate-buffered saline
[0262] EGTA--ethylene glycol-bis(.beta.-aminoethyl ether)
N,N,N',N'-tetraacetic acid
[0263] GDP--guanosine diphosphate
[0264] sc--subcutaneous
[0265] po--orally
[0266] ip--intraperitoneal
[0267] icv--intra cerebro ventricular
[0268] iv--intravenous
[0269] [.sup.3H]SR141716A--radiolabeled
N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H--
pyrazole-3-carboxamide hydrochloride available from Amersham
Biosciences, Piscataway, N.J.
[0270] [.sup.3H]CP-55940--radiolabled 5-(1,1-dimethyl
heptyl)-2-[5-hydroxy-2-(3-hydroxypropyl)-cyclohexyl]-phenol
available from NEN Life Science Products, Boston, Mass.
[0271]
AM251--N-(piperidin-1-yl)-1-(2,4-dichlorophenyl)-5-(4-iodophenyl)--
4-methyl-1H-pyrazole-3-carboxamide available from Tocris.TM.,
Ellisville, Mo.
[0272] Compounds having an activity <20 nM are generally tested
in the CB-1 GTP.gamma. [.sup.35S] Binding Assay and the CB-2
binding assay described below in the Biological Binding Assays
section. Selected compounds are then tested in vivo using one or
more of the functional assays described in the Biological
Functional Assays section below. CB-1 binding activities of 0.5-250
nM were observed for Examples 1-5.
In Vitro Biological Assays
[0273] Bioassay systems for determining the CB-1 and CB-2 binding
properties and pharmacological activity of cannabinoid receptor
ligands are described by Roger G. Pertwee in "Pharmacology of
Cannabinoid Receptor Ligands" Current Medicinal Chemistry, 6,
635-664 (1999) and in WO 92/02640 (U.S. application Ser. No.
07/564,075 filed Aug. 8, 1990, incorporated herein by
reference).
[0274] The following assays were designed to detect compounds that
inhibit the binding of [.sup.3H] SR141716A (selective radiolabeled
CB-1 ligand) and [.sup.3H]
5-(1,1-dimethylheptyl)-2-[5-hydroxy-2-(3-hydroxypropyl)-cyclohexyl]-pheno-
l; radiolabeled CB-1/CB-2 ligand) to their respective
receptors.
Rat CB-1 Receptor Binding Protocol
[0275] PelFreeze brains (available from Pel Freeze Biologicals,
Rogers, Ark.) were cut up and placed in tissue preparation buffer
(5 mM Tris HCl, pH=7.4 and 2 mM EDTA), polytroned at high speed and
kept on ice for 15 minutes. The homogenate was then spun at
1,000.times.g for 5 minutes at 4.degree. C. The supernatant was
recovered and centrifuged at 100,000.times.G for 1 hour at
4.degree. C. The pellet was then re-suspended in 25 ml of TME (25
nM Tris, pH=7.4, 5 mM MgCl.sub.2, and 1 mM EDTA) per brain used. A
protein assay was performed and 200 .mu.l of tissue totaling 20
.mu.g was added to the assay.
[0276] The test compounds were diluted in drug buffer (0.5% BSA,
10% DMSO and TME) and then 25 .mu.l were added to a deep well
polypropylene plate. [.sup.3H] SR141716A was diluted in a ligand
buffer (0.5% BSA plus TME) and 25 .mu.l were added to the plate. A
BCA protein assay was used to determine the appropriate tissue
concentration and then 200 .mu.l of rat brain tissue at the
appropriate concentration was added to the plate. The plates were
covered and placed in an incubator at 20.degree. C. for 60 minutes.
At the end of the incubation period 250 .mu.l of stop buffer (5%
BSA plus TME) was added to the reaction plate. The plates were then
harvested by Skatron onto GF/B filtermats presoaked in BSA (5
mg/ml) plus TME. Each filter was washed twice. The filters were
dried overnight. In the morning the filters were counted on a
Wallac Betaplate.TM. counter (available from PerkinElmer Life
Sciences.TM., Boston, Mass.).
Human CB-1 Receptor Binding Protocol
[0277] Human embryonic kidney 293 (HEK 293) cells transfected with
the CB-1 receptor cDNA (obtained from Dr. Debra Kendall, University
of Connecticut) were harvested in homogenization buffer (10 mM
EDTA, 10 mM EGTA, 10 mM Na Bicarbonate, protease inhibitors;
pH=7.4), and homogenized with a Dounce Homogenizer. The homogenate
was then spun at 1,000.times.g for 5 minutes at 4.degree. C. The
supernatant was recovered and centrifuged at 25,000.times.G for 20
minutes at 4.degree. C. The pellet was then re-suspended in 10 ml
of homogenization buffer and re-spun at 25,000.times.G for 20
minutes at 4.degree. C. The final pellet was re-suspended in 1 ml
of TME (25 mM Tris buffer (pH=7.4) containing 5 mM MgCl.sub.2 and 1
mM EDTA). A protein assay was performed and 200 .mu.l of tissue
totaling 20 .mu.g was added to the assay.
[0278] The test compounds were diluted in drug buffer (0.5% BSA,
10% DMSO and TME) and then 25 .mu.l were added to a deep well
polypropylene plate. [.sup.3H] SR141716A was diluted in a ligand
buffer (0.5% BSA plus TME) and 25 .mu.l were added to the plate.
The plates were covered and placed in an incubator at 30.degree. C.
for 60 minutes. At the end of the incubation period 250 .mu.l of
stop buffer (5% BSA plus TME) was added to the reaction plate. The
plates were then harvested by Skatron onto GF/B filtermats
presoaked in BSA (5 mg/ml) plus TME. Each filter was washed twice.
The filters were dried overnight. In the morning the filters were
counted on a Wallac Betaplate.TM. counter (available from
PerkinElmer Life Sciences.TM., Boston, Mass.).
CB-2 Receptor Binding Protocol
[0279] Chinese hamster ovary-K1 (CHO-K1) cells transfected with
CB-2 cDNA (obtained from Dr. Debra Kendall, University of
Connecticut) were harvested in tissue preparation buffer (5 mM
Tris-HCl buffer (pH=7.4) containing 2 mM EDTA), polytroned at high
speed and kept on ice for 15 minutes. The homogenate was then spun
at 1,000.times.g for 5 minutes at 4.degree. C. The supernatant was
recovered and centrifuged at 100,000.times.G for 1 hour at
4.degree. C. The pellet was then re-suspended in 25 ml of TME (25
mM Tris buffer (pH=7.4) containing 5 mM MgCl.sub.2 and 1 mM EDTA)
per brain used. A protein assay was performed and 200 .mu.l of
tissue totaling 10 .mu.g was added to the assay.
[0280] The test compounds were diluted in drug buffer (0.5% BSA,
10% DMSO, and 80.5% TME) and then 25 .mu.l were added to the deep
well polypropylene plate. [.sup.3H]
5-(1,1-Dimethyl-heptyl)-2-[5-hydroxy-2-(3-hydroxy-propyl)-cyclohexyl]-phe-
nol was diluted a ligand buffer (0.5% BSA and 99.5% TME) and then
25 .mu.l were added to each well at a concentration of 1 nM. A BCA
protein assay was used to determine the appropriate tissue
concentration and 200 .mu.l of the tissue at the appropriate
concentration was added to the plate. The plates were covered and
placed in an incubator at 30.degree. C. for 60 minutes. At the end
of the incubation period 250 .mu.l of stop buffer (5% BSA plus TME)
was added to the reaction plate. The plates were then harvested by
Skatron format onto GF/B filtermats presoaked in BSA (5 mg/ml) plus
TME. Each filter was washed twice. The filters were dried
overnight. The filters were then counted on the Wallac
Betaplate.TM. counter.
CB-1 GTP.gamma. [.sup.35S] Binding Assay
[0281] Membranes were prepared from CHO-K1 cells stably transfected
with the human CB-1 receptor cDNA. Membranes were prepared from
cells as described by Bass et al, in "Identification and
characterization of novel somatostatin antagonists," Molecular
Pharmacology, 50, 709-715 (1996). GTP.gamma. [.sup.35S] binding
assays were performed in a 96 well FlashPlate.TM. format in
duplicate using 100 pM GTP.gamma.[.sup.35S] and 10 .mu.g membrane
per well in assay buffer composed of 50 mM Tris HCl, pH 7.4, 3 mM
MgCl.sub.2, pH 7.4, 10 mM MgCl.sub.2, 20 mM EGTA, 100 mM NaCl, 30
.mu.M GDP, 0.1% bovine serum albumin and the following protease
inhibitors: 100 .mu.g/ml bacitracin, 100 .mu.g/ml benzamidine, 5
.mu.g/ml aprotinin, 5 .mu.g/ml leupeptin. The assay mix was then
incubated with increasing concentrations of antagonist (10.sup.-10
M to 10.sup.-5 M) for 10 minutes and challenged with the
cannabinoid agonist
5-(1,1-dimethyl-heptyl)-2-[5-hydroxy-2-(3-hydroxy-propyl)-cyclohexyl]-phe-
nol (10 .mu.M). Assays were performed at 30.degree. C. for one
hour. The FlashPlates.TM. were then centrifuged at 2000.times.g for
10 minutes. Stimulation of GTP.gamma.[.sup.35S] binding was then
quantified using a Wallac Microbeta.EC.sub.50 calculations done
using Prism.TM. by Graphpad.
[0282] Inverse agonism was measured in the absense of agonist.
CB-1 FLIPR-Based Functional Assay Protocol
[0283] CHO-K1 cells co-transfected with the human CB-1 receptor
cDNA (obtained from Dr. Debra Kendall, University of Connecticut)
and the promiscuous G-protein G16 were used for this assay. Cells
were plated 48 hours in advance at 12500 cells per well on collagen
coated 384 well black clear assay plates. Cells were incubated for
one hour with 4 .mu.M Fluo-4 AM (Molecular Probes) in DMEM (Gibco)
containing 2.5 mM probenicid and pluronic acid (0.04%). The plates
were then washed 3 times with HEPES-buffered saline (containing
probenicid; 2.5 mM) to remove excess dye. After 20 min the plates
were added to the FLIPR individually and fluorescence levels was
continuously monitored over an 80 second period. Compound additions
were made simultaneously to all 384 wells after 20 seconds of
baseline. Assays were performed in triplicate and 6 point
concentration-response curves generated. Antagonist compounds were
subsequently challenged with 3 .mu.M WIN 55,212-2 (agonist). Data
were analyzed using Graph Pad Prism.
Detection of Inverse Agonists
[0284] The following cyclic-AMP assay protocol using intact cells
was used to determine inverse agonist activity.
[0285] Cells were plated into a 96-well plate at a plating density
of 10,000 cells per well at a concentration of 100 .mu.l per well.
The plates were incubated for 24 hours in a 37.degree. C.
incubator. The media was removed and media lacking serum (100
.mu.l) was added. The plates were then incubated for 18 hours at
37.degree. C.
[0286] Serum free medium containing 1 mM IBMX was added to each
well followed by 10 .mu.l of test compound (1:10 stock solution (25
mM compound in DMSO) into 50% DMSO/PBS) diluted 10.times. in PBS
with 0.1% BSA. After incubating for 20 minutes at 37.degree. C., 2
.mu.M of Forskolin was added and then incubated for an additional
20 minutes at 37.degree. C. The media was removed, 100 .mu.l of
0.01N HCl was added and then incubated for 20 minutes at room
temperature. Cell lysate (75 .mu.l) along with 25 .mu.l of assay
buffer (supplied in FlashPlate.TM. cAMP assay kit available from
NEN Life Science Products Boston, Mass.) into a Flashplate. cAMP
standards and cAMP tracer were added following the kit's protocol.
The flashplate was then incubated for 18 hours at 4.degree. C. The
content of the wells were aspirated and counted in a Scintillation
counter.
In Vivo Biological Assays
[0287] Cannabinoid agoinists such as
.DELTA..sup.9-tetrahydrocannabinol (.DELTA..sup.9-THC) and
5-(1,1-dimethyl-heptyl)-2-[5-hydroxy-2-(3-hydroxy-propyl)-cyclohexyl]-phe-
nol have been shown to affect four characteristic behaviors in
mice, collectively known as the Tetrad. For a description of these
behaviors see: Smith, P. B., et al. in "The pharmacological
activity of anandamide, a putative endogenous cannabinoid, in
mice." J. Pharmacol. Exp. Ther., 270(1), 219-227 (1994) and Wiley,
J., et al. in "Discriminative stimulus effects of anandamide in
rats," Eur. J. Pharmacol., 276(I-2), 49-54 (1995). Reversal of
these activities in the Locomotor Activity, Catalepsy, Hypothermia,
and Hot Plate assays described below provides a screen for in vivo
activity of CB-1 antagonists.
[0288] All data is presented as % reversal from agonist alone using
the following formula:
(5-(1,1-dimethyl-heptyl)-2-[5-hydroxy-2-(3-hydroxy-propyl)-cyclohexyl]-ph-
enol/agonist-vehicle/agonist)/(vehicle/vehicle-vehicle/agonist).
Negative numbers indicate a potentiation of the agonist activity or
non-antagonist activity. Positive numbers indicate a reversal of
activity for that particular test.
Locomotor Activity
[0289] Male ICR mice (n=6; 17-19 g, Charles River Laboratories,
Inc., Wilmington, Mass.) were pre-treated with test compound (sc,
po, ip, or icv). Fifteen minutes later, the mice were challenged
with
5-(1,1-dimethyl-heptyl)-2-[5-hydroxy-2-(3-hydroxy-propyl)-cyclohexyl]-phe-
nol (sc). Twenty-five minutes after the agonist injection, the mice
were placed in clear acrylic cages (431.8 cm.times.20.9
cm.times.20.3 cm) containing clean wood shavings. The subjects were
allowed to explore surroundings for a total of about 5 minutes and
the activity was recorded by infrared motion detectors (available
from Coulbourn Instruments.TM., Allentown, Pa.) that were placed on
top of the cages. The data was computer collected and expressed as
"movement units."
Catalepsy
[0290] Male ICR mice (n=6; 17-19 g upon arrival) were pre-treated
with test compound (sc, po, ip or icv). Fifteen minutes later, the
mice were challenged with
5-(1,1-dimethyl-heptyl)-2-[5-hydroxy-2-(3-hydroxy-propyl)-cyclohexyl]-phe-
nol (sc). Ninety minutes post injection, the mice were placed on a
6.5 cm steel ring attached to a ring stand at a height of about 12
inches. The ring was mounted in a horizontal orientation and the
mouse was suspended in the gap of the ring with fore- and hind-paws
gripping the perimeter. The duration that the mouse remained
completely motionless (except for respiratory movements) was
recorded over a 3-minute period.
[0291] The data were presented as a percent immobility rating. The
rating was calculated by dividing the number of seconds the mouse
remains motionless by the total time of the observation period and
multiplying the result by 100. A percent reversal from the agonist
was then calculated.
Hypothermia
[0292] Male ICR mice (n=5; 17-19 g upon arrival) were pretreated
with test compounds (sc, po, ip or icv). Fifteen minutes later,
mice were challenged with the cannabinoid agonist
5-(1,1-dimethyl-heptyl)-2-[5-hydroxy-2-(3-hydroxy-propyl)-cyclohexyl]-phe-
nol (sc). Sixty-five minutes post agonist injection, rectal body
temperatures were taken. This was done by inserting a small
thermostat probe approximately 2-2.5 cm into the rectum.
Temperatures were recorded to the nearest tenth of a degree
Hot Plate
[0293] Male ICR mice (n=7; 17-19 g upon arrival) are pre-treated
with test compounds (sc, po, ip or iv). Fifteen minutes later, mice
were challenged with a cannabinoid agonist
5-(1,1-dimethyl-heptyl)-2-[5-hydroxy-2-(3-hydroxy-propyl)-cyclohexyl]-phe-
nol (sc). Forty-five minutes later, each mouse was tested for
reversal of analgesia using a standard hot plate meter (Columbus
Instruments). The hot plate was 10''.times.10''.times.0.75'' with a
surrounding clear acrylic wall. Latency to kick, lick or flick
hindpaw or jump from the platform was recorded to the nearest tenth
of a second. The timer was experimenter activated and each test had
a 40 second cut off. Data were presented as a percent reversal of
the agonist induced analgesia.
Food Intake
[0294] The following screen was used to evaluate the efficacy of
test compounds for inhibiting food intake in Sprague-Dawley rats
after an overnight fast.
[0295] Male Sprague-Dawley rats were obtained from Charles River
Laboratories, Inc. (Wilmington, Mass.). The rats were individually
housed and fed powdered chow. They were maintained on a 12-hour
light/dark cycle and received food and water ad libitum. The
animals were acclimated to the vivarium for a period of one week
before testing was conducted. Testing was completed during the
light portion of the cycle.
[0296] To conduct the food intake efficacy screen, rats were
transferred to individual test cages without food the afternoon
prior to testing, and the rats were fasted overnight. After the
overnight fast, rats were dosed the following morning with vehicle
or test compounds. A known antagonist was dosed (3 mg/kg) as a
positive control, and a control group received vehicle alone (no
compound). The test compounds were dosed at ranges between 0.1 and
100 mg/kg depending upon the compound. The standard vehicle was
0.5% (w/v) methylcellulose in water and the standard route of
administration was oral. However, different vehicles and routes of
administration were used to accommodate various compounds when
required. Food was provided to the rats 30 minutes after dosing and
the Oxymax automated food intake system (Columbus Instruments,
Columbus, Ohio) was started. Individual rat food intake was
recorded continuously at 10-minute intervals for a period of two
hours. When required, food intake was recorded manually using an
electronic scale; food was weighed every 30 minutes after food was
provided up to four hours after food was provided. Compound
efficacy was determined by comparing the food intake pattern of
compound-treated rats to vehicle and the standard positive
control.
Alcohol Intake
[0297] The following protocol evaluates the effects of alcohol
intake in alcohol preferring (P) female rats (bred at Indiana
University) with an extensive drinking history. The following
references provide detailed descriptions of P rats: Li, T.-K., et
al., "Indiana selection studies on alcohol related behaviors" in
Development of Animal Models as Pharmacogenetic Tools (eds McClearn
C. E., Deitrich R. A. and Erwin V. G.), Research Monograph 6,
171-192 (1981) NIAAA, ADAMHA, Rockville, Md.; Lumeng, L, et al.,
"New strains of rats with alcohol preference and nonpreference"
Alcohol And Aldehyde Metabolizing Systems, 3, Academic Press, New
York, 537-544 (1977); and Lumeng, L, et al., "Different
sensitivities to ethanol in alcohol-preferring and -nonpreferring
rats," Pharmacol, Biochem Behav., 16, 125-130 (1982).
[0298] Female rats were given 2 hours of access to alcohol (10% v/v
and water, 2-bottle choice) daily at the onset of the dark cycle.
The rats were maintained on a reverse cycle to facilitate
experimenter interactions. The animals were initially assigned to
four groups equated for alcohol intakes: Group 1--vehicle (n=8);
Group 2--positive control (e.g., 5.6 mg/kg AM251; n=8); Group
3--low dose test compound (n=8); and Group 4--high dose of test
compound (n=8). Test compounds were generally mixed into a vehicle
of 30% (w/v) .beta.-cyclodextrin in distilled water at a volume of
1-2 ml/kg. Vehicle injections were given to all groups for the
first two days of the experiment. This was followed by 2 days of
drug injections (to the appropriate groups) and a final day of
vehicle injections. On the drug injection days, drugs were given sc
30 minutes prior to a 2-hour alcohol access period. Alcohol intake
for all animals was measured during the test period and a
comparison was made between drug and vehicle-treated animals to
determine effects of the compounds on alcohol drinking
behavior.
[0299] Additional drinking studies were done utilizing female
C57Bl/6 mice (Charles River). Several studies have shown that this
strain of mice will readily consume alcohol with little to no
manipulation required (Middaugh et al., "Ethanol Consumption by
C57BL/6 Mice: Influence of Gender and Procedural Variables"
Alcohol, 17 (3), 175-183, 1999; Le et al., "Alcohol Consumption by
C57BL/6, BALA/c, and DBA/2 Mice in a Limited Access Paradigm"
Pharmacology Biochemistry and Behavior, 47, 375-378, 1994).
[0300] For our purposes, upon arrival (17-19 g) mice were
individually housed and given unlimited access to powdered rat
chow, water and a 10% (w/v) alcohol solution. After 2-3 weeks of
unlimited access, water was restricted for 20 hours and alcohol was
restricted to only 2 hours access daily. This was done in a manner
that the access period was the last 2 hours of the dark part of the
light cycle.
[0301] Once drinking behavior stabilized, testing commenced. Mice
were considered stable when the average alcohol consumption for 3
days was .+-.20% of the average for all 3 days. Day 1 of test
consisted of all mice receiving vehicle injection (sc or ip).
Thirty to 120 minutes post injection access was given to alcohol
and water. Alcohol consumption for that day was calculated (g/kg)
and groups were assigned (n=7-10) so that all groups had equivocal
alcohol intake. On day 2 and 3, mice were injected with vehicle or
drug and the same protocol as the previous day was followed. Day 4
was wash out and no injections were given. Data was analyzed using
repeated measures ANOVA. Change in water or alcohol consumption was
compared back to vehicle for each day of the test. Positive results
would be interpreted as a compound that was able to significantly
reduce alcohol consumption while having no effect on water
Oxygen Consumption
Methods:
[0302] Whole body oxygen consumption is measured using an indirect
calorimeter (Oxymax from Columbus Instruments, Columbus, Ohio) in
male Sprague Dawley rats (if another rat strain or female rats are
used, it will be specified). Rats (300-380 g body weight) are
placed in the calorimeter chambers and the chambers are placed in
activity monitors. These studies are done during the light cycle.
Prior to the measurement of oxygen consumption, the rats are fed
standard chow ad libitum. During the measurement of oxygen
consumption, food is not available. Basal pre-dose oxygen
consumption and ambulatory activity are measured every 10 minutes
for 2.5 to 3 hours. At the end of the basal pre-dosing period, the
chambers are opened and the animals are administered a single dose
of compound (the usual dose range is 0.001 to 10 mg/kg) by oral
gavage (or other route of administration as specified, i.e., sc,
ip, iv). Drugs are prepared in methylcellulose, water or other
specified vehicle (examples include PEG400, 30% beta-cyclo dextran
and propylene glycol). Oxygen consumption and ambulatory activity
are measured every 10 minutes for an additional 1-6 hours
post-dosing.
[0303] The Oxymax calorimeter software calculates the oxygen
consumption (ml/kg/h) based on the flow rate of air through the
chambers and difference in oxygen content at inlet and output
ports. The activity monitors have 15 infrared light beams spaced
one inch apart on each axis, ambulatory activity is recorded when
two consecutive beams are broken and the results are recorded as
counts.
[0304] Resting oxygen consumption, during pre- and post-dosing, is
calculated by averaging the 10-min O.sub.2 consumption values,
excluding periods of high ambulatory activity (ambulatory activity
count>100) and excluding the first 5 values of the pre-dose
period and the first value from the post-dose period. Change in
oxygen consumption is reported as percent and is calculated by
dividing the post-dosing resting oxygen consumption by the pre-dose
oxygen consumption*100. Experiments will typically be done with
n=4-6 rats and results reported are mean+/-SEM.
Interpretation:
[0305] An increase in oxygen consumption of >10% is considered a
positive result. Historically, vehicle-treated rats have no change
in oxygen consumption from pre-dose basal.
* * * * *