Cannabinoid receptor ligands and uses thereof

(54) Title: CANABINOID RECEPTOR BINDERS AND THEIR USES (30) Unionist Priority: 23/04/2003 us 60 / 464,831 (71) Depositor (s): Pfizer Products Inc. (US) (72) Inventor (s) : Philip Albert Carpino, Robert Lee Dow (74) Attorney: Dannemann, Siemsen, Bigler & Ipanema Moreira (86) International Request: pct ib2004 / 001482 of 20/04/2004 (87) International Publication: wo 2004/094429 of 04 / 11/2004 (57) Abstract: CANABINOID RECEPTOR BINDERS AND THEIR USES. There are described in this, compounds of Formula (I) and (II) that act as cannabinoid receptor ligands and their use in the treatment of disorders related to the cannabinoid receptor mediation in animals.

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(II)

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Invention Patent Descriptive Report for CANABINOID RECEPTOR BINDERS AND THEIR USES.

Field of the Invention

The present invention relates to pyrazolyl and imidazolyl compounds as cannabinoid receptor ligands, in particular CB1 receptor antagonists, and their uses for the treatment of diseases, conditions and / or disorders modulated by cannabinoid receptor antagonists.

Background

Obesity is a major public health concern due to its increasing prevalence and associated health risks. Obesity and overweight are generally defined by the body mass index (BMI or 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 / m²). Excess weight is typically defined as a BMI of 25-29.9 kg / m², and obesity is typically defined as a BMI of 30 kg / m². See, for example, National Heart, Lung and Blood Institute, Clinical Guidelines on the Identification, Evaluation and Treatment of Overweight and Obesity in Adults, The Evidence Report, Washington, DC: US ​​Department of Health and Human Services, NIH publication n ° 98 -4083 (1998).

The increase in obesity is worrying due to the excessive health risk associated with obesity, including coronary heart disease, strokes, hypertension, type 2 diabetes mellitus, dyslipidemia, sleep apnea, osteoarthritis, gallbladder disease, depression, and certain forms of cancer (for example, of the endometrium, breast, prostate and colon). The negative health consequences of obesity make it the second leading cause of preventable death in the United States of America and have a significant economic and psychosocial effect on society. See M. McGinnis, WH Foege, Actual Causes of Death in the United States in JAMA, 270 (1993) 2207-12.

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 treating obesity is to improve cardiovascular and metabolic values ​​to reduce obesity-related morbidity and mortality. It has been shown that the loss of 5-10% of body mass can substantially improve metabolic values, such as blood glucose, blood pressure and lipid concentrations. Therefore, it is believed that an intentional reduction of 5-10% of body mass can reduce morbidity and mortality.

Prescription drugs currently available for the treatment of obesity generally reduce weight by inducing satiety or decreasing the absorption of dietary fat. Satiety is achieved by increasing synaptic levels of norepinephrine, serotonin or both. For example, stimulation of serotonin receptors of subtypes 1B, 1D and 2C and adrenergic receptors 1 and 2 decreases food intake by regulating satiety. See GA Bray, The New Era of Drug Treatment. Pharmacologic Treatment of Obesity: Symposium OverView, Obes. Res., 2 (suppl 4) (1995) 415s-7s. Adrenergic agents (for example, diethylproponic acid, benzphetamine, phendimetrazine, mazindol and phentermine) act by modulating norepinephrine and dopamine receptors by promoting the release of catecholamine. Older adrenergic weight loss drugs (eg, amphetamine, methamphetamine and phenmetrazine), which take full advantage of dopamine's pathways, are no longer recommended due to the risk of their abuse. Fenfluramine and dexfenfluramine, both serotonergic agents used to regulate appetite, are no longer available for use.

More recently, CB1 cannabinoid receptor antagonists / inverse agonists have been suggested as potential appetite suppressants. See, for example, M. Arnone et al., Selective Inhibition of Sucrose and Ethanol Intake by SR141716, an Antagonist of Central Cannabinoid (CB1) Receptors, Psychopharmacol., 132 (1997) 104-106; G. Colombo et al., Appetite Supression and Weight Loss after the Cannabinoid Antagonist SR 141716, Life Sci., 63 (1998) PL113-PL117; J. Simiand et al., SR141716, CB1 Cannabinoid Receptor Antagonist, Selectively Reduces Sweet Food Intake in Marmose, Behav. Pharmacol., 9 (1998) 179-181; and F. Chaperon et al., Involvement of Central Cannabinoid (CB1) Receptors in the Establishment of Place Conditioning in Rats, Psychopharmacology, 135 (1998) 324-332. For a review of CB1 and CB2 cannabinoid receptor modulators, see RG Pertwee, Cannabinoid Receptor Ligands: Clinical and Neuropharmacological Considerations, Relevant to Future Drug Discovery and Development, Exp. Opin. Invest. Drugs, 9 (7) (2000) 1553-1571.

Although investigations are ongoing, there is still a need for a more effective and safer therapeutic treatment to reduce or prevent weight gain.

In addition to obesity, there is also an unmet need for the treatment of alcohol abuse. Alcoholism affects approximately 10.9 million men and 4.4 million women in the United States of America. Approximately 100,000 deaths per year have been attributed to alcohol abuse or dependence. The 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 the form of endogenous cannabinoid plays a critical role in controlling the entry of ethanol. The endogenous CB1 receptor antagonist SR141716A has been shown to block the voluntary entry of ethanol into rats and mice. See M. Arnone etal., Selective Inhibition of Sucrose and Ethanol Intake by SR141716, an Antagonist of Central Cannabinoid (CB1) Receptors, Psychopharmacol., 132 (1997) 104-106. For a review, see BL Hungund and BS Basavarajappa, Are Anadamide and Cannabinoid Receptors involved in Ethanol Tolerance? A Review of the Evidence, Alcohol &amp; Alcoholism, 35 (2) (2000) 126-133.

Current treatments for alcohol abuse or addiction currently suffer from non-adherence or potential hepatotoxicity; therefore, there is a high unmet need for more effective treatment of alcohol abuse / dependence.

summary

The present invention provides compounds of Formula (I) or (II) that act as cannabinoid receptor ligands (in particular, CB1 receptor antagonists)

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on what

A is nitrogen and B is carbon, or A is carbon and B is nitrogen;

R ° is an aryl optionally substituted with one or more substituents, or a heteroaryl optionally substituted with one or more substituents (preferably, R ° is a substituted phenyl, more preferably a phenyl substituted with one to three substituents independently selected from the group consisting of halo (preferably chlorine or fluorine), C1-C4 alkoxy, C1-C4 alkyl, halo-substituted C1-C4 alkyl (preferably fluorine-substituted alkyl) and cyano, and the most preferable is R¹ be 2chlorophenyl, 2-fluorophenyl, 2,4-dichlorophenyl, 2-fluor-4-chlorophenyl, 2-chloro-4fluorophenyl or 2,4-difluorophenyl);

R¹ is an aryl optionally substituted with one or more substituents, heteroaryl optionally substituted with one or more substituents, -CH = CH-, R^1a, or -CH₂CH₂-R^1a, where R^1a is hydrogen or a selected chemical portion of CrCe alkyl, 3- or 8-membered partially or completely saturated carbocyclic ring or rings, 3- or 6-membered partially or completely saturated heterocycle, aryl, heteroaryl, where the chemical moiety is optionally substituted with one or more substituents;

X is a bond or -C (R^2a) (R^2b) -; where R^2a and R^2b are each independently hydrogen, C1-C4 alkyl or halo-substituted C1-C4 alkyl (preferably R^2a and R^2b are both hydrogen);

R^3rd and R^3b are each independently hydrogen, C1-C4 alkyl or halo-substituted C1-C4 alkyl; and

R⁴ is a chemical portion selected from the group consisting of C 1 -C alkyl₈, aryl, heteroaryl, aryl-C1-C4 alkyl, 3 to 8 membered carbocyclic ring or rings partially or completely saturated, C1-C3 heteroarylalkyl, 5 to 6 membered latona, 5 to 6 membered lactam, and 3-heterocycle to 8 members partially or completely saturated, where said chemical moiety is optionally substituted with one or more substituents;

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.

In a preferred embodiment of the present invention, a compound of Formula (III) or (IV) is provided

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on what

A, Β, X, R^2a, R^2b, R^3rd, R^3b and R⁴ are as defined above;

R °, R °; R¹ and R¹ are each independently halo, C 1 -C alkoxy₄, C-C alkyl₄, halo-substituted C1-C4 alkyl, or cyano;

a pharmaceutically acceptable salt thereof, a prodrug of the compound or salt, or a solvate or hydrate of the compound, salt or prodrug.

In the preferred embodiments of the present invention, R⁴ is a chemical moiety selected from the group consisting of CrCs alkyl, aryl-C1-C4 alkyl, and 3 to 8 membered carbocyclic ring or rings partially or completely saturated, where said chemical moiety is optionally substituted with one or more substituents.

More preferably, R⁴ is CrCe alkyl, Ci-C alkyl₈ replaced by halo, cyclopentyl, cyclohexyl, piperidin-1-yl, pyrrolidin-1-yl or morpholin-1-yl.

Preferably, R ° and R¹ they are each independently, a phenyl substituted with 1 to 3 substituents independently selected from the group consisting of halo, C1-C4 alkoxy, C1-C4 alkyl, C1-C4 alkyl substituted by halo, and cyano;

more preferably, R ° and R¹ are each independently selected from the group consisting of chlorine, fluorine, C1-C4 alkoxy, C1C alkyl₄, fluorine-substituted C1-C4 alkyl, and cyano;

most preferably, R ° is 2-chlorophenyl, 2-fluorophenyl, 2,4dichlorophenyl, 2-fluor-4-chlorophenyl, 2-chloro-4-fluorophenyl, or 2,4-difluorophenyl; and R¹ be 4-chlorophenyl, 4-cyanophenyl or 4-fluorophenyl.

Preferred compounds of Formula (I) where A is nitrogen, B is carbon and X is a double bond include:

2- (2-chloro-phenyl) -5-isopropyl-3- (3,4,5-trifluoro-phenyl) -4,5-dihydro-2H-pyrrole [3,4c] pyrazol-6-one; 2,3-bis- (2-chloro-phenyl) -5-isopropyl-4,5-dihydro-2H-pyrrole [3,4c] pyrazol-6-one; 2- (2-chloro-phenyl) -5-isopropyl-3- (4-methoxymethyl-phenyl) -4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one; 2- (2-chloro-phenyl) -3- (2-fluoro-phenyl) -5isopropyl-4,5-dihydro-2H-pyrrol {3,4-c] pyrazol-6-one;

2- (2-chloro-phenyl) -5-isopropyl-3- (6-methoxy-pyridin-3-yl) -4,5-dihydro-2H

-pyrrole [3,4-c] pyrazol-6-one;

3- (3-chloro-4-fluoro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl-4,5-dihydro-2H-pyrrole [3,4c] pyrazole-6-one;

2- (2-chloro-phenyl) -3- (4-fluoro-3-methyl-phenyl) -5-isopropyl-4,5-dihydro-2H

-pyrrole [3,4-c] pyrazol-6-one;

3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl-4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one;

3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl-4-methyl-4,5-dihydro-2H-pyrrole [3,4c] pyrazol-6-one;

4-benzyl-3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl-4,5-dihydro2H-pyrrol [3,4-c] pyrazol-6-one;

5- tert-butyl-3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one;

3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-cyclobutyl-4,5-dihydro-2H-pyrrole [3,4-c] pyrazol-6-one;

3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-cyclopentyl-4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one;

3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-cyclohexyl-4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one;

3- (4-chloro-phenyl) -2- (2,4-dichloro-phenyl) -5-isopropyl-4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one;

5-tert-butyl-3- (4-chloro-phenyl) -2- (2,4-dichloro-phenyl) -4,5-dihydro-2H-pyrrol [3,4c] pyrazol-6-one;

3- (4-chloro-phenyl) -5-cyclopentyl-2- (2,4-dichloro-phenyl) -4,5-dihydro-2Hpirrol [3,4-c] pyrazol-6-one;

3- (2-chloro-phenyl) -2- (4-chloro-phenyl) -5-isopropyl-4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one; 3- (2-chloro-phenyl) -2- (4-chloro-phenyl) -5-cyclopentyl-4,5-dihydro2H-pyrrole [3,4-c] pyrazol-6-one;

3- (2-chloro-phenyl) -2- (4-chloro-phenyl) -5-cyclohexyl-4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one; 5-bicycle [2.2.1] hept-2-yl-3- (2-chloro-phenyl) -2- (4-chloro-phenyl) -4,5-dihydro

-2H-pyrrole [3,4-c] pyrazol-6-one;

2- (4-chloro-phenyl) -5-cyclopentyl-3- (2-fluoro-phenyl) -4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one; and

2- (4-chloro-phenyl) -5-cyclohexyl-3- (2-fluoro-phenyl) -4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one;

a pharmaceutically acceptable salt thereof, or a solvate or hydrate of said compound or said salt.

The most preferred compounds include:

3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl-4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one;

3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5- (2,2,2-trifluoro-ethyl) -4,5-dihydro2H-pyrrole [3,4-c] pyrazole- 6-one; and

4- [2- (2-chloro-phenyl) -5-isopropyl-6-oxo-2,4,5,6-tetrahydro-pyrrole [3,4-c] pyrazol-3-yl] -benzonitrile; a pharmaceutically acceptable salt thereof, or a solvate or hydrate of said compound or said salt.

Preferred compounds of Formula (I) where A is nitrogen, B is carbon and X is -C (R^2a) (R^2b) - include: 3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -6-isopropyl-2,4,5,6-tetrahydropyrazolo [3,4-c] pyridin-7-one; 3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -6- (2,2,2-trifluoroethyl) -2,4,5,6-tetrahydro-pyrazolo [3,4-c] pyridin-7-one; 3- (4-chloro-phenyl) -2- (2-chlorophenyl) -6- (2,2-difluoro-ethyl) -2,4,5,6-tetrahydro-pyrazolo [3,4-c] pyridin- 7-one; and

3- {4-chloro-phenyl) -2- (2-chloro-phenyl) -6- (2-fluoro-ethyl) -2,4,5,6-tetrahydropyrazolo [3,4-c] pyridin-7- ona; a pharmaceutically acceptable salt thereof, or a solvate or hydrate of the compound or salt.

Preferred compounds of Formula (I), where A is carbon, B is nitrogen and X is a bond, include:

2- (4-chloro-phenyl) -5-cyclopentyl-1- (2-fluoro-phenyl) -5,6-dihydro-1H-pyrrol [3,4-d] imidazole-4-one;

2- (4-chloro-phenyl) -5-cyclopentyl-1- (2,4-dichloro-phenyl) -5,6-dihydro-1H-pyrrol [3,4d] imidazole-4-one;

2- (4-chloro-phenyl) -5-cyclohexyl-1- (2,4-dichloro-phenyl) -5,6-dihydro-1Hpirrol [3,4-d] imidazole-4-one;

2- (4-chloro-phenyl) -1 - (2,4-dichloro-phenii) -5- (2,2,2-trifluoro-ethyl) -5,6-dihydro-1Hpirrol [3,4-d] imidazole-4-one;

2- (4-chloro-phenyl) -1- (2-chloro-phenyl) -5-isopropyl-5,6-dihydro-1H-pyrrol [3,4-d] imidazole-4-one;

2- (4-chloro-phenyl) -1- (2-chloro-phenyl) -5-cyclopentyl-5,6-dihydro-1H-pyrrol [3,4-d] imidazole-4-one;

2- (4-chloro-phenyl) -1- (2-chloro-phenyl) -5-cyclohexyl-5,6-dihydro-1H-pyrrol [3,4-d] imidazole-4-one;

2- (2-chloro-phenyl) -1 - (4-chloro-phenyl) -5-cyclohexyl-5,6-dihydro-1 H-pyrrol [3,4-d] imidazole-4-one;

2- (4-chloro-phenyl) -5-cyclohexylmethyl-1- (2-fluoro-phenyl) -5,6-dihydro-1 H-pyrrol [3,4-d] imidazole-4-one;

5-cyclopentyl-2- (4-fluoro-phenyl) -1- (2-fluoro-phenyl) -5,6-dihydro-1 H-pyrrol [3,4-d] imidazole-4-one;

1- (2-chloro-phenyl) -5-cyclopentyl-2- (4-fluoro-phenyl) -5,6-dihydro-1 H-pyrrol [3,4-d] imidazole-4-one;

2- (2-chloro-phenyl) -1- (4-chloro-phenyl) -5-cyclopentyl-5,6-dihydro-1 H-pyrrol [3,4-d] imidazole-4-one;

2- (2-chloro-phenii) -1- (4-chloro-phenyl) -5-cyclohexyl-5,6-dihydro-1 H-pyrrol [3,4-d] imidazole-4-one; and

1- (4-chloro-phenyl) -5-cyclohexyl-2- (2,4-dichloro-phenyl) -5,6-dihydro-1 H-pyrrol [3,4-d] imidazole-4-one ; a pharmaceutically acceptable salt thereof, or a solvate or hydrate of the compound or salt.

A more preferred compound is 2- (4-chloro-phenyl) -cyclopentyl-1- (2-fluoro-phenyl) -5,6-dihydro-1H-pyrrole [3,4-d] imidazole-4-one; a pharmaceutically acceptable salt thereof, or a solvate or hydrate of the compound or salt.

A preferred compound of Formula (I) where A is carbon, B is nitrogen and X is -C (R^2a) (R^2b) - is 2- (4-chloro-phenyl) -1- (2-chloro-phenyl) -5-cyclopentyl-1,5,6,7-tetrahydro-imidazo [4,5-c] pyridin-4-one ; a pharmaceutically acceptable salt thereof, or a solvate or hydrate of the compound or salt.

Preferred compounds of Formula (I) where R¹ is -CH = CH-R^1a include:

2- (2-chloro-phenyl) -3- [2- (4-chloro-phenyl) -vinyl] -5-isopropyl-4,5-dihydro-2H-pyrrole [3,4c] pyrazole-6-one;

2- (2-chloro-phenyl) -5-isopropyl-3-vinyl-4,5-dihydro-2H-pyrrole [3,4-c] pyrazol-6-one;

2- (2-chloro-phenyl) -3- [2- (4-chloro-phenyl) -vinyl] -5- (2,2,2-trifluoro-ethyl) -4,5-dihydro-2Hpirroi [3, 4-c] pyrazol-6-one;

2- (2-chloro-phenyl) -3- [2- (4-chloro-phenyl) -vinyl] -5- (2,2-difluoro-ethyl) -4,5-dihydro-2Hpirrol [3,4- c] pyrazol-6-one; and

2- (2-chloro-phenyl) -3- [2- (4-chloro-phenyl) -vinyl] -5- (2-fluoro-ethyl) -4,5-dihydro-2H-pyrrole [3,4- c] pyrazol-6-one; a pharmaceutically acceptable salt thereof, or a solvate or hydrate of the compound or salt.

Preferred compounds of Formula (II) include: 2- (4-chlorophenyl) -5-cyclohexyl-3- (2-fluoro-phenyl) -2,4,5,6-tetrahydro-pyrrole [3,4- c] pyrazole;

3- (4-chloro-phenyl) -5-cyclopentyl-2- (2,4-dichloro-phenyl) -2,4,5,6-tetrahydro-pyrrole [3,4cjpyazole; and

3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl-2,4,5,6-tetrahydro-pyrrole [3,4-pyrazole; a pharmaceutically acceptable salt thereof, or a solvate or hydrate of the compound or salt.

Some of the compounds described herein can contain at least one chiral center; accordingly, those skilled in the art will appreciate that all stereoisomers (e.g., enantiomers and diastereomers) of the compounds illustrated and discussed herein are within the scope and scope of the present invention. In addition, the tautomeric forms of the compounds are also within the scope and scope of the present invention. Those skilled in the art will recognize that chemical moieties, such as an alphaaminoether or alpha-chloroamine, may be too unstable to be isolated; therefore, such portions are not part of this invention.

The 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 comprising (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 can also contain at least one additional pharmaceutical agent (described herein). Preferred agents include partial nicotine receptor agonists, opioid antagonists (eg, naltrexone and nalmefene), dopaminergic agents (eg, apomorphine), agents for attention deficit disorder (ADD including attention deficit hyperactivity disorder ( ADHD)) (for example, Ritalin®, Strattera®, Concerta® and

Adderall®) and anti-obesity agents (described here below).

In yet another embodiment of the present invention, a method for treating a disease, condition or disorder modulated by a cannabinoid receptor antagonist (preferably, a CB1 receptor) in mammals that includes the step of administration to an animal with need for such treatment of a therapeutically effective amount of a compound of the present invention (or a pharmaceutical composition thereof).

Diseases, conditions and / or disorders modulated by cannabinoid receptor antagonists include eating disorders (eg, excessive drinking disorder, anorexia and bulimia), weight loss or control (eg, reduced calorie intake or and / or appetite suppression), obesity, depression, atypical depression, bipolar mood disorders, psychoses, schizophrenia, behavioral addictions, suppression of reward-related behaviors (eg avoidance of conditioned place, such as cocaine and morphine-induced suppression of conditioned place), substance abuse, addictive disorders, impulsivity, alcoholism (eg, alcohol abuse, addiction and / or addiction including withdrawal treatment, reducing cravings and preventing recurrence of alcohol intake, tobacco abuse (e.g., addiction, cessation and / or addiction to smoking, including craving treatment and prevention of relapse in tobacco smoking), dementia (including memory loss, Alzheimer's disease, elderly dementia, vascular dementia, mild cognitive impairment, age-related cognitive decline and mild neurocognitive disorder), male sexual dysfunction (e.g., difficulty with erection), seizures with seizures, epilepsy, inflammation, gastrointestinal disorders (e.g. dysfunction of gastrointestinal motility or intestinal propulsion), attention deficit disorder (ADI / ADHD), Parkinson's disease and type II diabetes. In a preferred embodiment, the method is used to treat weight loss, obesity, bulimia, ADD / ADHD, dementia, alcoholism and / or tobacco abuse.

The compounds of the present invention can be administered in combination with other pharmaceutical agents. Preferred pharmaceutical agents include partial nicotinic receptor agonists, opioid antagonists (eg, naltrexone (including naltrexone deposit), antabuse and nalmefene), dopaminergic agents (eg, apomorphine), ADD / ADHD agents (eg, hydrochloride methylphenidate (for example, Ritalin® and Concerta®), atomoxetine (for example, Strattera®) and amphetamines (for example, Adderall®)) and anti-obesity agents, such as apo-B / MTP inhibitors, 11β-hydroxysteroid-dehydrogenase-1 inhibitors (Ιΐβ-HSD type 1), YY3-36 peptide or its analogs, MCR-4 agonists, CCK-A agonists, monoamine reuptake inhibitors, sympathomimetic agents, adrenergic receptor agonists β3, dopamine receptor agonists, melanocyte stimulating hormone receptor analogs, 5-HT2c receptor agonists, melanin concentration hormone receptor antagonists, leptin, leptin analogs, leptin receptor agonists, galanin receptor antagonists, lipase inhibitors, bombesin receptor agonists, neuropeptide Y receptor antagonists (e.g., NPY Y5 receptor antagonists such as those described below), thyromimetic agents, dehydroepiandrosterone or its analogues, glucocorticoid receptor antagonists, orexin receptor antagonists, glucagon-type peptide receptor 1 agonists, ciliary neurotrophic factors, agouti-related human protein antagonists, ghrelin receptor antagonists, histamine receptor 3 antagonists or inverse agonists, and neuromedin U receptor agonists, and the like.

The combination therapy can be administered in the form of (a) a unique pharmaceutical composition comprising a compound of the present invention, at least one additional pharmaceutical agent described herein 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 herein and an excipient , diluent or pharmaceutically acceptable carrier. The pharmaceutical compositions can be administered simultaneously or sequentially, and in any order.

In yet another aspect of the present invention, a pharmaceutical kit is provided for use by a consumer to treat diseases, conditions or disorders modulated by cannabinoid receptor antagonists in an animal. The kit 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, conditions or disorders that are modulated by cannabinoid receptor antagonists (in particular, the CB1 receptor).

In yet another embodiment of the present invention, there 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 herein, and (ii) a pharmaceutically acceptable carrier, excipient or diluent; and c) a container.

Definitions

As used herein, the term alkyl refers to a hydrocarbon radical of the general formula C_nH2n + i · The alkane group can be linear or branched. For example, the term Οι-Οθ alkyl refers to a monovalent, linear or branched aliphatic group, containing from 1 to 6 carbon atoms (for example, methyl, ethyl, n-propyl, Apropila, n-butyl, Abutila, s-butyl, tbutyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, 3,3dimethylpropyl, hexyl, 2-methylpentyl, and the like). Similarly, the alkyl portion of an alkoxy group, acyl (e.g., alkanoyl), alkylamine, dialkylamine and alkylthia have the same definition as above. When indicated as being optionally substituted, the alkane radical or the 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 perchlor or perfluoroalkyls), independently selected from the group substituents listed below in the definition of substituted. Halo-substituted alkyl or halo-substituted alkyl refers to a group substituted with one or more halogen atoms (e.g., fluoromethyl, difluoromethyl, trifluoromethyl, perfluoroethyl and the like). When substituted, alkane radicals or alkyl moieties are preferably substituted with 1 to 3 fluorine substituents, or 1 or 2 substituents independently selected from C1-C3 alkyl, C cycloalkyl₃-Ç₆, C2-C3 alkenyl, aryl, heteroaryl, 3- to 6-membered heterocycle, chlorine, cyano, hydroxy, C1-O alkoxy₃, aryloxy, amine, (C1-C alkyl₆) amine, di (C1-C4 alkyl) amine, aminocarboxylate (i.e., (CrC3 alkyl) -OC (O) -NH-), hydroxy (C alkyl)₂Ç₃) amine or keto (oxo), and more preferably, 1 to 3 fluorine groups, or 1 substituent selected from C 1 -C alkyl₃, cycloalkyl C₃-Ç₆, arila C₆, 6-membered heteroaryl, 3 to 6-membered heterocycle, C 1 -C alkoxy₃, (C1C alkyl₄) amine or di (C 1 -C alkyl)₂)-the mine.

The terms partially or completely saturated carbocyclic ring (also referred to as partially or completely saturated cycloalkyl) refer to non-aromatic rings that are either partially or completely hydrogenated and can exist in the form of a single ring, bicyclic ring or spiral ring. Unless otherwise specified, the carbocyclic ring is generally a 3- to 8-membered ring. For example, partially or completely saturated carbocyclic (or cycloalkyl) rings include groups such as cyclopropyl, cyclopropenyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, norbornyl (bicyclo [2.2.1] heptila, nortilyl, nortilyl, nortilyl, nortilyl), bicycles [2.2.2] octyl and the like. When designated as being optionally substituted, the partially saturated or fully saturated cycloalkyl group can 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 in which the carbocyclic ring is fused with a phenyl ring (for example, indanyl).

The carbocyclic group can be linked to a chemical entity or moiety by any 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 C1-C3 alkyl, C2-C3 alkenyl, C-C alkylidenyl₆, aryl, heteroaryl, 3- to 6-membered heterocycle, chlorine, fluorine, cyano, hydroxy, C1-C3 alkoxy, aryloxy, amine, (C1C6 alkyl) amine, di (C-C4 alkyl) -amine, aminocarboxylate (i.e. (C 1 -C alkyl₃) -OC (O) -NH-), hydroxy (C2-C3 alkyl) amine or keto (oxo), and more preferably, 1 or 2 substituents independently selected from C1-C2 alkyl, 3- to 6-membered heterocycle, fluorine , C1-C3 alkoxy, (C1-C alkyl₄) amine or di (C alkyl)₁-Ç₂)-the mine. Similarly, any cycloalkyl portion of a group (for example, cycloalkylalkyl, cycloalkylamino, etc.) has the same definition as above.

The term partially saturated or completely saturated heterocyclic ring (also referred to as partially saturated or completely saturated heterocycle) refers to non-aromatic rings that are either partially or completely hydrogenated and can exist in the form of a single ring, bicyclic ring or spiral ring . Unless otherwise specified, the heterocyclic ring is generally a 3 to 6 membered ring containing 1 to 3 hetero atoms (preferably 1 or 2 hetero atoms) 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, piperinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, 2 oxazinyl, morpholine, thiomorpholine, tetrahydrothienyl, tetrahydrothienyl-1,1-dioxide and the like. When indicated as being optionally substituted, the partially saturated or completely saturated heterocycle group can 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 heterocyclic ring also includes groups in which the heterocyclic ring is fused with 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 C1-C3 alkyl, C cycloalkyl₃-Ç₆, C2-C4 alkenyl, aryl, heteroaryl, 3- to 6-membered heterocycle, chlorine, fluorine, cyano, hydroxy, Ci-C alkoxy₃, aryloxy, amine, (C1-C alkyl₆) amine, di (C 1 -C alkyl)₃) -amine, aminocarboxylate (i.e., (C1-C alkyl)₃) OC (O) -NH-), or keto (oxo), and more preferably, with 1 or 2 substituents independently selected from C 1 -C alkyl₃, cycloalkyl C₃-C6, aryl C₆, 6-membered heteroaryl, 3 to 6-membered heterocycle or fluorine. The heterocyclic group can be attached to the chemical entity or moiety by any of the ring atoms within the system in the heterocyclic ring. Similarly, any heterocycle portion of a group (e.g., heterocycle-substituted alkyl, heterocycle-carbonyl, etc.) has the same definition as above.

The term aryl or aromatic carbocyclic ring refers to aromatic moieties having a single ring (e.g., phenyl) or fused (e.g. naphthalene, anthracene, phenanthrene, etc.) system. A typical aryl group is a 6- to 10-membered aromatic carbocyclic ring or rings. When indicated as being optionally substituted, aryl groups can be unsubstituted or substituted with one or more substituents (preferably, not more than three substituents) independently selected from the group of substituents listed below in the definition for substituted. The substituted aryl groups include a chain of aromatic moieties (for example, biphenyl, terphenyl, phenylnaphthalyl, etc.). When substituted, the aromatic moieties are preferably substituted with 1 or 2 substituents independently selected from C1-C4 alkyl, C alkenyl₂-Ç₃, aryl, heteroaryl, 3- to 6-membered heterocycle, bromine, chlorine, fluorine, iodine, cyano, hydroxy, C1-C4 alkoxy, aryloxy, amine, (C 1 -C 6 alkyl) amine, di (C 1 -C alkyl)₃) -amine or aminocarboxylate (i.e., (C 1 -C alkyl)₃) -OC (O) -NH-), and more preferably, 1 or 2 substituents independently selected from C1-C4 alkyl, chlorine, fluorine, cyano, hydroxy or C1-C4 alkoxy. The aryl group can be linked to the chemical entity or moiety by any of the carbon atoms within the system in the aromatic ring. Similarly, the aryl portion (i.e., the aromatic portion) of an aroyl or aroyloxy (i.e., (aryl) C (O) -O-) has the same definition as above.

The term heteroaryl or heteroaromatic ring refers to aromatic moieties containing at least one heteroatom (for example, oxygen, sulfur, nitrogen or their combinations) within a 5 to 10-membered aromatic ring system (for example, pyrrolyl, pyridyl, pyrazolyl, indolyl, indazolyl, thienyl, furanyl, benzofuranyl, oxazolyl, imidazolyl, tetrazolyl, triazinyl, pyrimidyl, pyrazinyl, thiazolyl, purinyl, benzimidazolyl, quinolinyl, isoquinolinyl, benzothiophenyl, benzoxazolyl, etc.). The heteroaromatic portion 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, heteroaryl groups can be unsubstituted or substituted with one or more substituents (preferably, not 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 C1-C4 alkyl, C alkenyl₂-Ç₃, aryl, heteroaryl, 3- to 6-membered heterocycle, bromine, chlorine, fluorine, iodine, cyano, hydroxy, C1-C4 alkoxy, aryloxy, amine, (C1-C alkyl)₆) amine, di (CrC alkyl)₃) -amine or aminocarboxylate (i.e., (C 1 -C alkyl)₃) -OC (O) -NH-), and more preferably, 1 or 2 substituents independently selected from C1-C4 alkyl, chlorine, fluorine, cyano, hydroxy, C1-C4 alkoxy, (C1 -C6 alkyl) amine or di (C1-alkyl) -Ç₃)-the mine. The heteroaryl group can be linked to the chemical entity or moiety by any of the atoms within the system in the aromatic ring (for example, 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 portion) of a heteroaryl or heteroaryloxy (i.e., (heteroaryl) -C (O) -O-) has the same definition as above.

The term acyl refers to hydrogen, alkyl groups, partially saturated or completely saturated cycloalkyl, partially saturated or completely saturated heterocycle, aryl and carbonyl substituted by heteroaryl. For example, acyl includes groups such as C 1 -C alkanoyl₆(for example, formyl, acetyl, propionyl, butyryl, valeryl, caproil, tbutilacetyl, etc.), (cycloalkyl C₃-Ç₆) carbonyl (e.g., cyclopropylcarbonyl, cyclobutylcarbonyl, cyclopentylcarbonyl, cyclohexylcarbonyl, etc.), heterocyclic carbonyl (e.g., pyrrolidinylcarbonyl, pyrrolid-2-one-5carbonyl, piperidinylcarbonyl, piperazinylcarbonyl, tetra-hydrocarbonyl, tetra-hydrocarbon, etc.) , benzoyl) and heteroaryl (for example, thiophenyl-2-carbonyl, thiophenyl-3-carbonyl, furanyl-2-carbonyl, furanyl-3-carbonyl, 1Hpirrol-2-carbonyl, 1H-pyrrole-3-carbonyl, benzo [b] thiophenyl-2-carbonyl, etc.). In addition, the alkyl, cycloalkyl, heterocycle, aryl and heteroaryl moiety of the acyl group can be any of the groups described in the respective definitions above. When indicated as being optionally substituted, the acyl group can 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 or the alkyl, cycloalkyl, heterocycle moiety, aryl and heteroaryl of the acyl group can be substituted as described above in the preferred and most preferred list of substituents, respectively.

The substituted term specifically foresees and allows one or more substitutions that are common in the art. However, it is generally understood by those skilled in the art that substituents should be selected in such a way that they do not adversely affect the pharmacological characteristics of the compound or interfere adversely with the use of the drug. Suitable substituents for any of the groups defined above include C 1 -C alkyl₆, cycloalkyl C₃-Ç₇, alkenyl C₂-Ç₆, Ci-C alkylidenyl₆, aryl, heteroaryl, 3- to 6-membered heterocycle, halo (for example, chlorine, bromine, iodine and fluorine), cyano, hydroxy, Ci-C alkoxy₆, aryloxy, sulfhydryl (mercapto), (C1-C alkyl)₆) thio, arylthio, amine, mono- or di (C 1 -C alkyl)₆) -amine, quaternary ammonium salts, Ci-C amine-alkoxy₆, aminocarboxylate (i.e., (CrCe alkyl) OC (O) -NH-), hydroxy (C alkyl)₂-C6) amine, amine (C1-C alkyl)₆) uncle, cyanoamino, nitro, C-C carbamyl₆₎ keto (oxo), acyl, (C 1 -C alkyl)₆) -CO2-, glycolyl, glycyl, hydrazine, guanyl, sulfamyl, sulfonyl, sulfinyl, uncle (C1 -C6 alkyl) -C (O) -, uncle (C1 -C6 alkyl) -CO2- and their combinations. In the case of substituted combinations, such as substituted ArC-aryl alkyl, either the aryl group or the alkyl group can be substituted, or both aryl and alkyl groups can be substituted with one or more substituents (typically, one to three substituents, except in the case of per-halo substitutions). A carbocyclic or heterocyclic group substituted by aryl or heteroaryl may be a fused ring (for example, indanyl, dihydrobenzofuranyl, dihydroindolyl, etc.).

The term solvate refers to a molecular complex of a compound represented by Formula (I) or (II) (including prodrugs and their pharmaceutically acceptable salts) with one or more solvent molecules. Such solvent molecules are those commonly used in the pharmaceutical art, which are known to be harmless to the recipient, for example, water, ethanol and the like. The term hydrate refers to the complex where the solvent molecule is water.

The term protecting group or Pg refers to a substituent that is commonly used to block or protect a particular functionality while other functional groups in the compound react. For example, an amine protecting group is a substituent attached to an amine group that blocks or protects the amine functionality in the compound. Suitable amine protecting groups include acetyl, trifluoroacetyl, tbutoxycarbonyl (BOC), benzyloxycarbonyl (CBz) and 9-fluorenylmethyleneoxycarbonyl (Fmoc). Similarly, a hydroxy protecting group refers to a substituent of a hydroxy group that blocks or protects 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 -CH₂CH₂SO2Ph, cyanoethyl, 2- (trimethylsilyl) ethyl, 2 (trimethylsilyl) ethoxymethyl, 2- (p-toluene-sulfonyl) ethyl, 2- (p-nitrophenylsulfenyl) ethyl, 2- (diphenylphosphino) ethyl, nitroethyl and the like. For a general description of protection groups and their use, see T. W. Greene, Protective Groups in Organic Synthesis, John Wiley &amp; Sons, New York, 1991.

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) alleviates, improves 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.

The term animal refers to humans (man or woman), companion animals (for example, dogs, cats and horses), food source animals, zoo animals, marine animals, birds and other similar animal species. Edible animals refers to animals that are food sources such as cows, pigs, sheep and poultry.

The pharmaceutically acceptable phrase indicates that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients comprising a formulation, and / or the mammal to be treated with them.

The terms treating, treating or treatment encompasses not only preventive, that is, prophylactic treatment, but also palliative treatment.

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 can act as an agonist, partial agonist, inverse agonist, antagonist or partial antagonist.

The term antagonist includes not only complete antagonists but also partial antagonists, as well as inverse agonists.

The term CB-1 receptor refers to the type 1 cannabinoid receptor coupled to the G protein.

The term compounds of the present invention (unless specifically identified otherwise) refers to compounds of Formulas (I), (II), (III) and (IV), their prodrugs, 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.

As used herein, structures drawn with circles within a ring designate aromaticity. For example, the following chemical portion designates a pyrazole ring when A is nitrogen and B is carbon; and the chemical portion designates an imidazole when A is a carbon and N is a nitrogen.

<img file="BRPI0409701A_D0006.tif" />

Detailed Description

The present invention provides compounds and their pharmaceutical formulations that are useful in the treatment of diseases, conditions and / or disorders modulated by cannabinoid receptor antagonists.

The compounds of the present invention can be synthesized by synthetic routes that include processes analogous to those well known in chemical techniques, particularly in light of the description contained herein. The starting materials are generally available from commercial sources such as Aldrich Chemicals (Milwaukee, Wl) or are readily prepared using methods well known to those skilled in the art (for example, prepared by methods generally described in Louis F. Fieser and Mary Fieser, Reagents for Organic Synthesis, ed. 1967-1999, Wiley, New York, Vol. 1-19, or Beilsteins Handbuch der organischen Chemie, 4, Aufl. ed. Springer-Verlag, Berlin, including supplements (also available via Beilstein online database)).

For illustrative purposes, the reaction schemes outlined below provide potential pathways for the synthesis of 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 can be used to synthesize the compounds of the invention. Although specific starting materials and reagents are indicated in the diagrams and discussed below, other starting materials and reagents can be easily replaced to produce a variety of derivatives and / or reaction conditions. In addition, many of the compounds prepared by the methods described below can still be modified in light of this disclosure, using conventional chemistry well known to those skilled in the art.

In the preparation of compounds of the present invention, protection of remote functionality (e.g., primary or secondary amine) from 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 amine protecting groups (NH-Pg) include acetyl, trifluoroacetyl, f-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 protection groups and their use, see TW Greene, Protective Groups in Organic Synthesis, John Wiley &amp; Sons, New York, 1991.

Scheme I outlines the general procedures that can be used to produce the compounds of the present invention where A is nitrogen, B is carbon, X is a bond and R^3rd and R^3b they are both hydrogen.

Scheme I

Η _O/^Ns + RO.CR ° NH₂²

CO₂R

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<img file="BRPI0409701A_D0008.tif" />

<img file="BRPI0409701A_D0009.tif" />

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<img file="BRPI0409701A_D0011.tif" />

The pyrazole intermediate l-1a can be prepared by cyclizing the desired hydrazine with acetylene dicarboxylate in the presence of a weak base (eg, alkali metal carbonate, such as potassium carbonate) in a protic solvent (eg, ethanol), under reflux conditions. Intermediate l-1a can then be produced by treating the pyrazole intermediate l-1a with phosphoryl bromide in the presence of dimethylformamide (DMF), in an aprotic solvent (eg 1,2dichloroethane), with heating. The amine group (R⁴-NH) can then be introduced into the molecule by treating the formyl compound l-1b with the desired amine (R⁴-NH) in the presence of sodium triacetoxy hydroxyborate and a weak acid (eg acetic acid). A variety of suitable amine compounds are either commercially available or easily synthesized using procedures well documented in the literature. The lactam can then be formed by hydrolysis of the pendant ester group and condensation of the pendant amine group with the carboxylic acid function, to form the amide chain. The lactam formation can be carried out using procedures well known to those skilled in the art. For example, the intermediate carboxylate ester 1-1c can be hydrolyzed using a strong base (for example, alkali metal hydroxide) in a polar solvent (for example, ethanol) with heating. The amide chain can then be formed by treating the resulting carboxylic acid with 1-propanophosphoric acid cyclic anhydride in the presence of a non-reactive base (e.g., triethylamine). Finally, the group R¹ is introduced into the molecule by displacing the bromine group with the R group¹ wanted. This can be achieved by treating the bromine intermediate l-1d with or the desired boronic acid (R¹-B (OH) 2) or the tin reagent (R¹SnR3), in the presence of cesium fluoride and tetracis (triphenylphosphine) palladium (0) in a polar solvent (1,2-dimethoxyethane), at elevated temperatures (for example, 100 ° C).

Alternatively, the compounds of the present invention where A is nitrogen, B is carbon and X is a bond, can be prepared using the procedures outlined below in Scheme II. Scheme II also illustrates the introduction of a vinyl portion in the R position¹, which can also be modified by reducing olefin functionality.

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<img file="BRPI0409701A_D0014.tif" />

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<img file="BRPI0409701A_D0016.tif" />

In Scheme II, the R portion¹ is introduced earlier in the synthetic scheme. Using the same basic procedures described above for displacing the bromine group in intermediate l-1d. intermediate l · 1b can be treated with the appropriate boronic acid, or tin reagent, to produce intermediate l-2a. The desired amine functionality can then be introduced, followed by hydrolysis to the corresponding carboxylic acid, and then cyclization to the lactam using the same procedures discussed above in Scheme I. When R¹ of the compound 1a is a vinyl group, the compound can be further modified by reacting the vinyl group with the desired aryl halide (for example, bromide or iodide) or heteroaryl halide (for example, bromide or iodide) in the presence of acetate palladium. Compound 1B can be further modified by reducing the double bond in the pendant group (R^1a-CH = CH-) of compound 1B using standard reduction techniques, such as those described in J. Amer. Chem. Soc., 91 (1969) 5769. For example, compound 1B is refluxed in 2-ethoxyethanol in the presence of p-toluenesulfonyl hydrazine.

Scheme III below illustrates the preparation of compounds of this invention where A is nitrogen, B is carbon and X is - (C (R^2a) (R^2b))26

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<img file="BRPI0409701A_D0021.tif" />

An extra methylene is introduced into the molecule by first reacting intermediate l-2a with (2- (trimethylsilyl) ethoxymethyl) triphenylphosphonium in the presence of sodium hydride to form the siloxy intermediate l · 3a. The siloxy group can then be removed by treating intermediate l-3a with a strong acid (e.g., hydrofluoric acid). The desired amine functionality can then be introduced by treating intermediate l-3b with the appropriate amine (R⁴NH₂), using procedures discussed above (for example, treatment with sodium triacethoxy hydrohydroborate and acetic acid in 1,2-dichloroethane). Cyclization to the lactam can be achieved by first hydrolysing the ester to the carboxylic acid and then cycling to the lactam using procedures similar to those discussed above (for example, (1) treatment with 1-propanophosphoric acid cyclic anhydride and triethylamine in dichloromethane ).

Scheme IV provides an alternative pathway for the synthesis of compounds of the present invention where A is nitrogen, B is carbon and X is a bond.

<img file="BRPI0409701A_D0022.tif" />

<img file="BRPI0409701A_D0023.tif" />

<img file="BRPI0409701A_D0024.tif" />

Starting material 1-4a can be prepared using procedures described by Barth et al., In European Patent Application EP 656354. The halo group (eg, bromine) can be introduced into the pending methyl group using procedures analogous to those described by Barth et al., in PCT Application WO 97/19063. For example, starting material 1-4a can be treated with 2,2'-azobisisobutyronitrile (AIBN) in carbon tetrachloride at elevated temperatures. The bromine group at 1-4b can then be displaced with the desired amine functionality using the same general procedures discussed above. Compound 1A can be formed by first hydrolyzing the ester group of Mç, followed by the formation of the amide chain using the general procedures discussed above. The compound lA can be further modified by the bonding of one or two groups pending on the carbon adjacent to the lactam nitrogen by treating the compound lA with the desired reagents (R^2a-L and / or R²-L, where L is a leaving group, such as a halo group (e.g., bromine)) in the presence of potassium hexamethyldisilazotide (KHMDSi) as described in Tet. Lett., 39 (1998) 2319-2320.

Scheme V illustrates a synthetic path for the preparation of compounds of the present invention where A is carbon, B is nitrogen and X is a bond, as well as the introduction of R^3rd and / or R^3b.

Scheme V

<img file="BRPI0409701A_D0025.tif" />

(lG)

Intermediate l-5a is prepared by treating the appropriate amine having the group R¹ with trimethylaluminium, under conditions of inert atmosphere, followed by condensation with the appropriate cyanide having the desired group R °. Suitable amines include substituted phenylamines (for example, 4-chlorophenylamine, 4-fluorophenylamine, 4-bromophenylamine, 4-phenophenylamine, 4-cyanophenylamine and the like), pyridin-2-ylamine, pyridin-3ylamine, pyridin-4-ylamine, substituted pyridinylamines for example, 2dimethylaminopyridin-5-ylamine, 2-methoxypyridin-5-ylamine, 5-chloropyrimine din-2-ylamine, 5-methylpyridin-2-yl, 5-methoxypyridin-2-ylamine, 3-chloropyridin-429 ylamine 2-N-morpholinylpyridin-5-yl and the like) and other commercially available or easily synthesized substituted or unsubstituted aryls and heteroarylamines. Suitable cyano compounds include substituted benzonitriles (for example, 2-chlorobenzonitrile, 2-fluorobenzonitrile, 2-methoxy benzonitrile, 2-methylbenzonitrile, 2,4-dichlorobenzonitrile, 2,4-difluorobenzonitrile, 2-chloro-4-fluorobenzonitrile, 2-chloro-4-chloro-benzyl 2,4-dimethoxybenzonitrile, 2-methyl-4-chlorobenzonitrile and the like), cyan-substituted pyridines (e.g. 4-cyano3-chloropyridine) and other commercially available or easily synthesized substituted or unsubstituted aryls and heteroarylnitriles.

Intermediate 1-5a can then be condensed with an ester of 3-bromo-2-oxo-propionic acid to produce the cyclized 4-hydroxy-4,5-dihydro-1H-imidazole ester 1-5b, using procedures similar to those described by IK Hhanna etal., J. Med. Chem., 40 (1997) 1634. For example, the intermediate amidine l-5a is refluxed in a polar solvent (for example, isopropanol) in the presence of a moderate base (for example, bicarbonate sodium). Generally, the reaction (i.e., cyclization followed by dehydration) proceeds directly to the desired imidazole ester intermediate 1-5c. In some cases, it may be necessary to dehydrate the initial carbinol condensation product 1-5b with an acid catalyst (e.g., toluenesulfonic acid in refluxing toluene), to provide the desired imidazole ester 1-5c.

The l-5c imidazole ester is prepared from the 4-hydroxy-4,5-dihydro l-5b intermediate using standard dehydration procedures well known to those skilled in the art. For example, intermediate 1-5b can be treated with p-toluenesulfonic acid monohydrate in refluxing toluene. Alternatively, intermediate 1-5b can be treated with methanesulfonyl chloride, in the presence of a base (e.g., triethylamine). The bromine group can be introduced to intermediate 1-5c by treatment with bromine following a procedure described in J. Het. Chem., 34 (3) (1997) 765-771. The bromine group at l-5c can then be converted to a formyl group to produce intermediate l-5d, first treating intermediate l-5c with a strong base (for example, n-butyl lithium), followed by DMF treatment. The amine functionality can then be introduced using procedures analogous to those discussed above for pyrazole compounds. For example, intermediate l-5d can react with the desired amine (R⁴-NH₂), in the presence of NaBH (OAc)₃, to produce intermediate l-5e. The amine intermediate 1-5e can then be cyclized to form the lactam by first hydrolyzing the ester group to its corresponding carboxylic acid followed by the formation of the amide chain using procedures analogous to those discussed above for pyrazole derivatives. The compound 1F can also be modified by linking the R groups^3rd and / or R^3b pending procedures similar to those discussed above. For example, treatment of compound lF with the desired reagents (R^2a-L and / or R^2aL, where L is a leaving group, such as a halo group (e.g., bromine)), in the presence of a base such as KHMDSi.

Scheme VI illustrates the preparation of compounds of the present invention where A is carbon, B is nitrogen (imidazole) and X is -C (R^2a) (R^2b)-.

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<img file="BRPI0409701A_D0027.tif" />

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<img file="BRPI0409701A_D0030.tif" />

<img file="BRPI0409701A_D0031.tif" />

The 1H imidazole compound can be prepared using procedures analogous to those discussed above for the preparation of the pyrazole derivative (ED). An extra methylene is introduced into the molecule by first reacting intermediate l-5e with the ylide formed from lithium hexamethyldisilazide and triphenylphosphonic (methoxymethyl) chloride to produce l-6a vinyl ether. The vinyl ether is converted to the corresponding aldehyde by heating the intermediate vinyl ether in an acidic environment. The amine functionality (R⁴NH) can then be introduced and the lactam ring formed using procedures analogous to those discussed above. As discussed above, the lactam can be formed by first hydrolyzing the ester, followed by the formation of the amide chain to produce the EH compound.

The compounds of Formula I, in which R⁴ is a substituted piperidinyl or pyrrolidinyl group, can be prepared as shown in Scheme VII.

Scheme VII

<img file="BRPI0409701A_D0032.tif" />

(lE) (ii) (lj)

Removal of the protecting group from compound 1E can be accomplished by methods known in the art to give bicyclic amine derivatives, such as Ei. which can subsequently react with alkyl halides in the presence of a suitable base, such as potassium carbonate in a solvent such as DMF or treated with acid chlorides or sulfonyl chlorides in the presence of a base, such as triethylamine, in a non-solvent polar, such as CH2CH2, to provide compounds such as 1J. The compound E1 can also react with an aldehyde derivative or ketone, in the presence of a reducing agent such as NaBH (OAc)₃, as previously described, to introduce the intermediate EJ · The compounds of Formulas ll-A and Scheme VIII.

Il-B can be prepared as shown in

Scheme VIII

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<img file="BRPI0409701A_D0034.tif" />

<img file="BRPI0409701A_D0035.tif" />

<img file="BRPI0409701A_D0036.tif" />

The pyrazole compound 1E is treated with an appropriate reducing agent such as lithium tetrahydridealuminate or borane (BH₃), in a polar aprotic solvent, such as THF, at temperatures in the range of about 0 ° C to about 100 ° C, to give compounds such as 11-A. Compound II-B can be prepared from ID using a similar reduction procedure. The imidazole compounds of formula II (A is carbon and

B is nitrogen) can be prepared using analogous procedures.

Conventional methods and / or techniques of separation and purification known to one skilled in the art can be used to isolate the compounds of the present invention, as well as their various related intermediates. Such techniques will be well known to one skilled 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 layer chromatography) thin), recrystallization and differential extraction techniques (ie, liquid-liquid).

The compounds of the present invention can be isolated and used on their own or in the form of their pharmaceutically acceptable salt, solvate and / or hydrate. The term salts refers to inorganic and organic salts of a compound of the present invention. Such 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, iodide, sulfate, bisulfate, nitrate, acetate, trifluoroacetate, oxalate, besylate, palmitate, pamoate, malonate, stearate salts, laurate, malate, borate, benzoate, lactate, phosphate, hexanofluorophosphate, hexanofluorosulfate , format, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glycoheptonate, latobionate and lauryl sulfonate, and the like. These may include cations based on 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, for example, Berge et al., J. Pharm. Sci., 66 (1977) 1-19.

The term prodrug means a compound that is transformed in vivo to produce a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate of the compound. Transformation can occur by several mechanisms, such as through hydrolysis in the blood. A discussion of the use of pro-drugs is provided by T. Higuchi and W. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the ACS Symposium Series and in Bioreversible Carriers in Drug Design, Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press, 1987.

For example, if a compound of the present invention contains a carboxylic acid functional group, a prodrug may contain an ester formed by replacing the hydrogen atom of the acid group with a group such as C 1 -C alkyl₈, (alkanoyl C₂-Ci2) oxymethyl, 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, Λ / (alkoxycarbonyl) aminomethyl having from 3 to 9 carbon atoms, 1- (N (alkoxycarbonyl) amine) ethyl having from 4 to 10 carbon atoms, 3-phthalidyl, 4-crotonolatonyl, gamma-butyrolaton-4-yl, di-N, N- (C 1 -C alkyl₂) amine-C alkyl₂-Ç₃ (such as β-dimethylaminoethyl), carbamoyl- (CrC alkyl₂), N, N-di (CrC alkyl ^ carbamoyl-CrC alkyl₂ and piperidino-, pyrrolidine- or morpholine (C alkyl₂-Ç₃).

Similarly, if a compound of the present invention contains an alcohol functional group, a prodrug can be formed by replacing the hydrogen atom of the alcohol group with a group such as (C 1 -C alkanoyl)₆) oxymethyl, 1 - ((Ci-C alkanoyl₆) oxy) ethyl, 1-methyl-1 - ((alkanoyl C_rC6) oxy) ethyl, (Ci-C alkoxy₆) carbonyloxymethyl, N- (Cr C alkoxy₆) carbonylaminomethyl, suctionyl, Ci-C alkanoyl₆, a-amine (Cr C alkanoyl₄), arylacyl and α-aminoacyl or α-aminoacyl-a-aminoacyl, where each aaminoacyl group is independently selected from the naturally occurring L-amino acids, P (O) (OH)₂, -P (O) (O (C 1 -C alkyl)₆)) 2 or glycosyl (the radical resulting from the removal of a hydroxyl group from the hemiacetal form of a carbohydrate).

If a compound of the present invention incorporates an amine functional group, a prodrug can be formed by replacing a hydrogen atom in the amine group with a group such as R-carbonyl, ROcarbonyl, NRR'-carbonyl where R and R 'are , each independently C1-C10 alkyl, C cycloalkyl₃-Ç₇, benzyl, or R-carbonyl is a natural α-aminoacyl or natural α-aminoacyl-natural a-aminoacyl, -C (OH) C (O) OY 'where Y' is H, C 1 -C alkyl₆ or benzyl, -C (OYo) Yi where Y_O is C alkyl_rÇ₄ and Yi is CrC alkyl₆, carboxy (C1-C alkyl)₆), amine (C alkyl)_rÇ₄) or mono-N- or diN, N- (C 1 -C alkyl)₆) aminoalkyl, -C (Y₂) Y₃ where Y₂ is H or methyl and Y₃ is mono-N- or di-N, N- (C 1 -C alkyl₆) amine, morpholine, piperidin-1-yl or pyrrolidin35

-ila.

The compounds of the present invention can 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 their mixtures, including racemic mixtures, form part of the present invention. In addition, the present invention contemplates all geometric and positional isomers. For example, if a compound of the present invention incorporates a double bond or a fused ring, both cis and trans forms, as well as mixtures thereof, are within the scope and scope of the invention.

Diastereomeric mixtures can be separated into their individual diastereoisomers on the basis of their physico-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 to a diastereomeric mixture by reacting with an appropriate optically active compound (eg, chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separation of diastereomers and conversion (eg, hydrolysis) from the individual diastereoisomers to the corresponding pure enantiomers. Also, some of the compounds of the present invention can be atropisomers (for example, substituted biaryls) and are considered to be part of this invention. The enantiomers can also be separated using a chiral HPLC column.

The compounds of the present invention can exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents, such as water, ethanol and the like, and the invention is intended to cover both solvated and unsolvated forms.

It is also possible that the intermediates and compounds of the present invention may exist in different tautomeric forms, and all of these forms are within the scope of the invention. The term tautomer or tautomeric form refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine isomerizations. A specific example of a proton tautomer is the imidazole portion, where the proton can migrate between the two ring nitrogen. Valency tautomers include interconversions by reorganizing some of the bonding electrons.

The present invention also encompasses isotopically labeled compounds of the present invention, which are identical to those cited herein, except that one or more atoms are exchanged for an atom that has an atomic mass or mass number different from the atomic mass or number of mass usually found in nature. Examples of isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine and chlorine, such as ²H, ³H, ¹¹Ç, ¹³Ç, ¹⁴Ç, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³¹P, ³²P, ³⁵S, ¹⁸F, ¹²³l, ¹²⁵le ³⁶CI, respectively.

Certain isotopically labeled compounds of the present invention (e.g., those labeled with ³H and ¹⁴C) are useful in distribution tests, in the tissue, of compound and / or substrate. Tritiated isotopes (ie,³H) and carbon-14 (ie, ¹⁴C) are particularly preferred for their ease of preparation and detectability. In addition, replacement with heavier isotopes such as deuterium (ie,²H) can produce certain therapeutic advantages resulting from increased metabolic stability (for example, increased in vivo half-life or reduced dosage requirements) and, therefore, may be preferred in some circumstances. Isotopes that emit positrons such as¹⁵O, ¹³N, ¹¹C e ¹⁸F are useful for positron emission tomography (PET) studies to examine receptor occupation on the substrate. The isotopically labeled compounds of the present invention can generally be prepared following procedures analogous to those disclosed in the Schemes and / or the Examples below, replacing an isotopically unlabeled reagent with an isotopically labeled reagent.

The compounds of the present invention are useful for the treatment of diseases, conditions and / or 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.

A typical formulation is prepared by mixing a compound of the present invention and a vehicle, diluent or excipient. Suitable vehicles, diluents and excipients are well known to those skilled in the art and include materials such as carbohydrates, waxes, water-soluble and / or expandable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water and the like. The particular vehicle, diluent or excipient used will depend on the medium and purpose for which the compound of the present invention is being applied. Solvents are generally selected on the basis of 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, which 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 can also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opacifying agents, glidants, processing aids, dyes, sweeteners, perfumers, agents flavorings and other known additives to provide an elegant presentation of the drug (ie a compound of the present invention or its pharmaceutical composition) or assist in the preparation of the pharmaceutical product (i.e., medicine).

Formulations can be prepared using conventional dissolving and mixing procedures. For example, the bulk of the drug substance (i.e., compound of the present invention or stabilized form of the compound (e.g., complex with a dextrin derivative or other known complexing agent)) is dissolved in a suitable solvent, in the presence of one or more of the excipients described above. The compound of the present invention is typically formulated in pharmaceutical dosage forms to provide an easily controllable dosage of the drug and to give the patient an elegant and easily handled product.

The pharmaceutical composition (or formulation) for application can be packaged in a variety of ways depending on the method used for administering the drug. Generally, an article for distribution includes a container having the pharmaceutical formulation deposited therein 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 assembly to prevent accidental access to the contents of the package. In addition, the container has a label on it that describes the contents of the container. The label can also include appropriate warnings.

The present invention further provides a method of treating diseases, conditions and / or disorders modulated by cannabinoid receptor antagonists in an animal which includes administration to an animal, which requires such treatment of a therapeutically effective amount of a compound of the present invention or a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present invention and an excipient, pharmaceutically acceptable diluent or carrier. The method is particularly useful for the treatment of diseases, conditions and / or disorders modulated by cannabinoid receptor antagonists (in particular, CB1 receptor).

Preliminary investigations have indicated that the following diseases, conditions and / or disorders are modulated by cannabinoid receptor antagonists: eating disorders (eg, drinking disorder with excessive drinking, anorexia and bulimia), weight loss or control (eg, reduced calorie or food intake, and / or appetite suppression), obesity, depression, depression atypical, bipolar mood disorders, psychoses, schizophrenia, behavioral addictions, suppression of behavior related to rewards (eg avoidance of conditioned place, such as cocaine and morphine-induced suppression of conditioned place preference), substance abuse, addictive disorders, impulsivity, alcoholism (for example, alcohol abuse, addiction and / or addiction, including abstinence treatment, craving reduction and prevention of recurrence of alcohol intake), tobacco abuse (e.g., addiction, cessation and / or addiction to smoking, including treatment of craving and prevention of relapse in tobacco smoking), dementia (including memory loss, Alzheimer's disease, dementia of the elderly, vascular dementia, mild cognitive impairment, age-related cognitive decline and mild neurocognitive disorder), male sexual dysfunction (eg, erection difficulty), attack disorders, epilepsy, inflammation, gastrointestinal disorders (for example, gastrointestinal motility dysfunction or intestinal propulsion), attention deficit disorder (ADD including attention deficit hyperactivity disorder (ADHD)), Parkinson's disease and type II diabetes.

Accordingly, the compounds of the present invention described herein are useful in the treatment of diseases, conditions or disorders that are modulated by cannabinoid receptor antagonists. Consequently, the compounds of the present invention (including the compositions and processes used herein) can be useful in the preparation of a medicament for the therapeutic applications described herein.

Other diseases, conditions and / or disorders for which cannabinoid receptor antagonists may be effective include: premenstrual syndrome or luteal phase syndrome, migraines, panic disorder, anxiety, post-traumatic syndrome, social phobia, weakening or cognitive disorder in non-demented individuals, mild non-amnesic cognitive weakness, postoperative cognitive decline, disorders associated with impulsive behaviors (such as disruptive behavior disorders (for example, anxiety / depression, improved executive function, tic disorders, conduct disorder and / or defiant opposition disorder), personality disorders in the adult (eg, borderline personality disorder and antisocial personality disorder), diseases associated with impulsive behaviors (eg substance abuse, paraphilia and self-mutilation) ) and impulsive control disorders (for example, intermittent explosive disorder, kleptomania, pyromania, pathological gambling and trichotillomania)), obsessive-compulsive disorder, chronic fatigue syndrome, male sexual dysfunction (eg, premature ejaculation), female sexual dysfunction, sleep disorders (eg, sleep apnea), autism, mutism, neurodegenerative movement disorders, spinal cord injury, nervous system injury central nervous system (e.g. trauma), stroke, neurodegenerative diseases or toxic or infectious CNS diseases (e.g. encephalitis or meningitis), cardiovascular disorders (e.g. thrombosis) and diabetes.

The compounds of the present invention can be administered to a patient at dosage levels in the range of about 0.7 mg to about 7000 mg per day. For a normal adult human having a body mass of about 70 kg, a dosage in the range of about 0.01 to about 100 mg per kilogram of body weight is typically sufficient. However, some variability in the general dosage range may be required, depending on the age and weight of the subject to be treated, the intended route of administration, the particular compound to be administered, and the like. Determination of dosage ranges and optimal dosages for a particular patient is well within the ability of one skilled in the art having the benefit of the present 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 well known to a person skilled in the art.

The compounds of this invention can also be used in conjunction with other pharmaceutical agents for the treatment of diseases, conditions and / or disorders described herein. Therefore, methods of treatment are also provided which include the administration of compounds of the present invention in combination with other pharmaceutical agents. Suitable pharmaceutical agents, which can be used in combination with the compounds of the present invention, include anti-obesity agents, such as apolipoprotein-B secretion inhibitors / microsomal triglyceride transfer protein (apo-B / MTP), 11β-hydroxysteroid inhibitors -deshhydrogenase-l (Ιΐβ-HSD type 1), YY3-36 peptide or its analogs, MCR-4 agonists, cholecystokinin-A (CCKA) agonists, monoamine reuptake inhibitors (such as sibutramine), sympathomimetic agents, β-adrenergic receptor agonists₃, dopamine agonists (such as bromocriptine), melanocyte stimulating hormone receptor analogs, 5-HT2c agonists, melanin concentration hormone receptor antagonists, leptin (the OB protein), leptin analogs, receptor agonists leptin, galanine antagonists, lipase inhibitors (such as tetrahydrolipstatin, i.e., orlistat), anoretic agents (such as a bombesin agonist), neuropeptide Y antagonists (for example, NPY Y5 receptor antagonists, such as the spiro-compounds described in US Patent Nos. 6,666,367, 6,649,624, 6,638,942, 6,605 720, 6 495 559, 6 462 053, 6 388 077, 6 335 345 and 6 326 375, US Publication Nos. 2002/0151456 and 2003/036652; and PCT Publication Nos. WO 03/010175, WO 03/082190 and WO 02/048152), thyromimetic agents, dehydroepiandrosterone or an analog thereof, glucocorticoid receptor agonists or antagonists, orexin receptor antagonists, glucagon-type peptide receptor 1 agonists, neurotrophic factors ciliary (such as Axokine® available from Regeneron Pharmaceuticals, Inc., Tarrytown, NY and Procter &amp; Gamble Company, Cincinnati, OH), agouti-related human protein (AGRP) antagonists, ghrelin receptor antagonists, histamine receptor 3 antagonists or inverse agonists, neuromedin U receptor agonists, and the like. Other anti-obesity agents, including the preferred agents listed below, are well known, or will become readily apparent in light of the present disclosure, to one skilled in the art.

Especially preferred are the antiobesity agents selected from the group consisting of orlistat, sibutramine, bromocriptine, ephedrine, leptin, pseudoephedrine and YY3-36 peptide or its analogs and 2-oxo-N- (5phenylpyrazinyl) spiro- [isobenzofuran-1 (3H), 4'-piperidine] -1 '-carboxamide. Preferably, the compounds of the present invention and combination therapies are administered in conjunction with exercise and a sensitive diet.

Representative anti-obesity agents for use in the combinations, pharmaceutical compositions and methods of the invention can be prepared using methods known to one skilled in the art, for example, sibutramine can be prepared as described in US Patent No. 4,929,629; bromocriptine can be prepared as described in US Patent No. 3,752,814 and 3,752,888; orlistat can be prepared as described in US Patent No. 5,274,143, 5,420,305, 5,540,917 and 5,643,874; PYY₃ -36 (including analogs) can be prepared as described in US Publication No. 2002/0141985 and WO 03/027637; and the NPY Y5 receptor antagonist 2-oxo-N- (5-phenylpyrazinyl) spiro- [isobenzofuran-1 (3H), 4'piperidine-Γ-carboxamide can be prepared as described in Publication US 002/0151456. Other NPY Y5 receptor antagonists include those described in PCT Publication No. 03/082190, such as

3-oxo-N- (5-phenyl-2-pyrazinyl) -spiro [isobenzofuran-1 (3H), 4'-piperidine] -carboxamide;

3-oxo-N- (7-trifluoromethylpyrido [3,2-b] pyridin-2-yl) -spiro- [isobenzofuran1 (3H), 4'-piperidine] -1-carboxamide;

N- [5- (3-fluorophenyl) -2-pyrimidinyl] -3-oxospiro- [isobenzofuran-1 (3H), [4'pi peridine] -1 '-carboxamide;

trans-3'-oxo-N- (5-phenyl-2-pyrimidinyl)] spiro [cyclohexane-1, r (3'H) isobenzofuran] -4-carboxamide;

trans-3'-oxo-N- [1 - (3-quinolyl) -4-imidazolyl] spiro [cyclohexane-1,1 '(3Ή) isobenzofuran] -4-carboxamide; trans-3-oxo-N- (5-phenyl-2-pyrazinyl) spiro [4azaiso-benzofuran-1 (3H), r-cyclohexane] -4'-carboxamide;

trans-N- [5- (3-fluorophenyl) -2-pyrimidinyl] -3-oxospiro [5-azaisobenzofuran1 (3H), 1'-cyclohexane] -4'-carboxamide;

trans-N- [5- (2-fluorophenyl) -2-pyrimidinyl] -3-oxospiro [5-azaisobenzofuran43

1 (3H), r-cyclohexane] -4'-carboxamide; trans-N- [1- (3,5-difluorophenyl) -4imidazolyl] -3-oxospiro [7-azaisobenzofuran-1 (3H), 1'-cyclohexanej-boxcarboxamide;

trans-3-oxo-N- (1-phenyl-4-pyrazolyl) spiro [4-azaisobenzofuran-1 (3H), 1'-cyclohexane] -4'-carboxamide;

trans-N- [1- (2-fluorophenyl) -3-pyrazolyl] -3-oxospiro [6-azaisobenzofuran1 (3H), 1'-cyclohexane] -4'-carboxamide;

trans-3-oxo-N- (1-phenyl-3-pyrazolyl) spiro [6-azaisobenzofuran-1 (3H), r-cyclohexane] -4'-carboxamide;

trans-3-oxo-N- (2-phenyl-1,2,3-triazol-4-yl) spiro [6-azaisobenzofuran-1 (3H), Tcyclohexane] -4'-carboxamide; and their pharmaceutically acceptable salts and esters. All of the above US Patents and Publications are hereby incorporated by reference.

Other suitable pharmaceutical agents that can be administered in combination with the compounds of the present invention include agents designed to treat tobacco abuse (e.g., partial nicotine receptor agonists, bupropion hypochloride (also known under the trademark Zyban®) therapies replacement drugs), agents to treat erectile dysfunction (for example, dopaminergic agents, such as apomorphine), DDA / ADHD agents (for example, Ritalin®, Strattera®, Concerta® and Adderall®) and agents to treat alcoholism, such as opioid antagonists (for example, naltrexone (also known under the trademark ReVia®) and nalmefene), disulfiram (also known under the trademark Antabuse®) and acamprosate (also known under the trademark Campral®)). In addition, agents for reducing alcohol withdrawal symptoms may also be co-administered, such as benzodiazepines, beta-blockers, clonidine, carbamazepine, pregabalin and gabapentin (Neurontin®). Treatment for alcoholism is preferably administered in combination with behavioral therapy, including components such as motivational stress therapy, cognitive behavioral therapy and guidance from self-help groups, including

Alcoholics Anonymous (AA).

Other pharmaceutical agents that may be useful include antihypertensive agents; anti-inflammatory agents (for example, COX-2 inhibitors); antidepressants (for example, fluoxetine hydrochloride (Prozac®)); cognitive enhancing agents (for example, donepezil hydrochloride (Aircept®) and other acetylcholinesterase inhibitors); neuroprotective agents (for example, memantine); antipsychotic medications (for example, ziprasidone (Geodon®), risperidone (Risperdal®) and olanzapine (Zyprexa®)); insulin and insulin analogs (for example, LysPro insulin); GLP-1 (7-37) (insulinotropin) and GLP-1 (7-36) -NH₂; sulfonylureas and their analogs: chlorpropamide, glibenclamide, tolbutamide, tolazamide, acetohexamide, Glypizide®, glimepiride, repaglinide, meglitinide; biguanides: metformin, phenformin, buformin; a2-antagonists and imidazolines: midaglizole, isaglidol, deriglidol, idazoxane, efaroxane, fluparoxane; other insulin secretors: linogliride, A-4166; glitazones: ciglitazone, Atos® (pioglitazone), englitazone, troglitazone, darglitazone, Avandia® (BRL49653); fatty acid oxidation inhibitors: clomoxir, etomoxir; α-glucosidase inhibitors: acarbose, miglitol, emiglitate, voglibose, MDL-25 637, camiglibose, MDL-73 945; βagonists: 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 (for example, Naglivan®) and peroxovanadium complexes; amylin antagonists; glucagon antagonists; gluconeogenesis inhibitors; somatostatin analogues; antilipolytic agents: nicotinic acid, acipimox, WAG 994, pramlintide (Symlin®), AC 2993, nateglinide, aldose-glutase inhibitors (eg zopolrestat), glycogen-phosphorylase inhibitors, sorbitoldeshydrogenase inhibitors, sodium-hydrogen exchanger type 1 inhibitors (NHE-1) and / or cholesterol biosynthesis inhibitors or cholesterol absorption inhibitors, especially an HMG-CoA reductase inhibitor, or an HMG-CoA synthase inhibitor, or an HMG-CoAreductase or synthase gene expression inhibitor, a CETP inhibitor, a bile acid scavenger, a fibrate, an ACAT inhibitor; a squalene synthase inhibitor, an anti-oxidant, or niacin. The compounds of the present invention can 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, gardic extract, extracts from the Hoodia plant, and niacin.

The dosage of the additional pharmaceutical agent generally depends on numerous factors, including the health of the individual being treated, the extent of treatment desired, the nature and type of concurrent therapy, if any, and the frequency of treatment and the nature of the desired effect. In general, the dosage range of the additional pharmaceutical agent is in the range of about 0.001 mg to about 100 mg per kilogram of body weight of the individual per day, preferably from about 0.1 mg to about 10 mg per kilogram of individual's body weight per day. However, some variability in the general dosage range may also be required depending on the age and weight of the subject to be treated, the intended route of administration, the particular anti-obesity agent to be administered and the like. Determining the optimal dosage ranges and dosages for a particular patient is well within the ability of one skilled in the art, having the benefit of the present disclosure.

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 an individual 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 (e.g., anti-obesity agent, partial nicotine receptor agonist, dopaminergic agent or opioid antagonist) can be administered either separately or in the pharmaceutical composition comprising both. It is generally preferred that such administration is oral. However, if the individual to be treated is unable to swallow or oral administration is otherwise harmful or undesirable, parenteral or transdermal administration may be appropriate. According to the methods of the invention, when a combination of a compound of the present invention and at least one other pharmaceutical agent is 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 is oral. It is especially preferred that such administration is oral and simultaneous. When a compound of the present invention and the additional pharmaceutical agent are administered sequentially, each can be administered by the same method or by different methods.

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 (e.g., intravenous, intramuscular or subcutaneous), intracisternal, intravaginal dosage form. , intraperitoneal, intravesical, local (for example, powder, ointment or drop), or buccal, or nasal.

Compositions suitable for parenteral injection generally include pharmaceutically acceptable sterile aqueous and non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous vehicles or diluents (including solvents and vehicles) include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol and the like), their suitable mixtures, vegetable oils (such as olive oil) and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by using a coating such as lecithin, by maintaining the required particle size in the case of dispersions, and by using surfactants.

These compositions can also contain excipients, such as preserving agents, humectants, emulsifiers and dispersants. The prevention of microorganism contamination of the compositions can be ensured by the inclusion of 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. The prolonged absorption of injectable pharmaceutical compositions can be caused by the use of agents capable of delaying absorption, for example, aluminum monostearate and gelatin.

Solid dosage forms for oral administration include capsules, tablets, powders and granules. In such solid dosage forms, a compound or combination of the present invention is mixed with at least one excipient, diluent or inert carrier. Suitable excipients, diluents or vehicles include materials such as sodium citrate or dicalcium phosphate or (a) fillers or fillers (e.g., starches, latose, sucrose, mannitol, silicic acid and the like); (b) binders (for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, acacia and the like); (c) humectants (for example, glycerol and the like); (d) disintegrating agents (for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, sodium carbonate and the like); (e) solution retarding agents (for example, paraffin and the like); (f) absorption accelerators (for example, quaternary ammonium compounds and the like); (g) wetting agents (for example, cetyl alcohol, glycerol monostearate and the like); (h) adsorbents (for example, 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 can also comprise buffering agents. Solid compositions of a similar type can also be used as fillers in soft or hard gelatin capsules using excipients such as latose or milk sugars, as well as high molecular weight polyethylene glycols, and the like.

Solid dosage forms, such as tablets, pills, capsules and granules, can be prepared with coatings and shells, such as enteric coatings and others well known in the art. They can also contain opacifying agents and can also be of a composition that they release the compound of the present invention and / or the additional pharmaceutical agent in a delayed manner. Examples of plug-in compositions that can be used are polymeric substances and waxes. The drug can also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the compound or combination of the present invention, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate , ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (for example, cotton oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, and the like), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid sorbitan esters, or mixtures of these substances, and the like.

In addition to such inert diluents, the composition may also include excipients such as wetting agents, emulsifying and suspending agents, sweeteners, flavorings and perfuming agents.

The suspensions, in addition to the compound of the present invention or combination, may further comprise vehicles such as suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum meta-hydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances, and the like.

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 vehicles, such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at ordinary ambient temperatures, but liquid at body temperature and therefore fuse in the rectum or vaginal cavity thereby releasing the active component (s).

Dosage forms for topical administration of the compounds and combinations of the compounds of the present invention with anti-obesity agents can comprise ointments, powders, sprays and inhalants. The drugs are mixed under sterile conditions with a pharmaceutically acceptable excipient, diluent or carrier, and any preservatives, buffers or propellants that may be required. Ophthalmic formulations, ointments, powders and eye solutions are also intended to be included within the scope of the present invention.

The following paragraphs describe formulations, dosages, etc. specimens, 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 carried out orally or non-orally (for example, by injection).

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, which is administered orally to an animal, is between about 0.01 and about 1000 mg / kg of body weight, preferably between about 0.01 and about 300 mg / kg of body weight.

Conveniently, a compound of the present invention (or combination) can be loaded into the drinking water so that a therapeutic dosage of the compound is ingested with the daily water supply. The compound can be directly measured for drinking water, preferably in the form of a liquid, water-soluble concentrate (such as an aqueous solution of a water-soluble salt).

Conveniently, a compound of the present invention (or combination) can also be added directly to the food, as such, or in the form of an animal food supplement, also referred to as a premix or concentrate. A premix or concentrate of the compound in an excipient, diluent or vehicle is most commonly employed for including the agent in the feed. Suitable carriers are liquid or solid, as desired, such as water, various flours such as alfalfa flour, soy flour, cottonseed oil meal, flax seed oil meal, corn cob meal and flour corn, molasses, urea, bone meal and mineral mixtures such as those commonly used in poultry feed. A particularly effective vehicle is the respective animal food itself; that is, a small portion of such food. The vehicle facilitates uniform distribution of the compound in the finished food, with which the premix is ​​mixed. Preferably, the compound is thoroughly mixed in the premix and, subsequently, in the food. In this regard, the compound can 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 mixed with the vehicle. It will be appreciated that the proportions of the compound in the concentrate are capable of wide variation since the amount of the compound in the finished food can be adjusted by mixing the appropriate proportion of the premix with the food to obtain a desired level of compound.

High power concentrates can be mixed by the food manufacturer with a protein carrier, such as soybean oil flour and other flours, as described above, to produce concentrated supplements that are suitable for direct animal feed. In such cases, animals are allowed to consume the usual diet. Alternatively, such concentrated supplements can be added directly to the food to produce a finished, nutritionally balanced food containing a therapeutically effective level of a compound of the present invention. The mixtures are thoroughly mixed by standard procedures, such as in a double jacket mixer, to ensure homogeneity.

If the supplement is used as a surface coating for the food, it likewise helps to ensure uniform distribution of the compound across the surface of the coated food.

Drinking water and effective food for increasing lean meat deposition and increasing 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 '³ up to 500 ppm of the compound in the food or water.

Preferred medicated pig, cattle, sheep and goat feeds 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 such animals being usually about 50 to 300 grams per ton of food. Food for poultry and pet animals usually contains from about 1 to about 400 grams and preferably from about 10 to 400 grams of a compound of the present invention (or combination) per ton of food.

For parenteral administration in animals, the compounds of the present invention (or combination) can be prepared in the form of a paste or a pellet and administered in the form of an implant, usually under the skin of the animal's head or ear, by which it is sought to increase deposition of lean meat and improvement in the ratio of lean meat to fat.

In general, parenteral administration involves injecting 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 the drug's body weight. The preferred dosage for poultry, pigs, cattle, sheep, goats and domestic pets is in the range of about 0.05 to about 10 mg / kg / day of drug body weight.

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.

Pellets containing an effective amount of a compound of the present invention, pharmaceutical composition, or combination, can be prepared by mixing a compound of the present invention or combination with a diluent such as "Carbowax", carnauba wax and the like, and a lubricant , such as magnesium or calcium stearate, can be added to improve the pelleting process.

It is certainly recognized that more than one pellet can be administered to an animal to achieve the desired dose level, which will provide an increase in lean meat deposition and an improvement in lean to fat ratio. In addition, implants can also be made periodically during the animal's treatment period in order to maintain the appropriate drug level in the animal's body.

The present invention has several advantageous veterinary characteristics. For the pet owner or veterinarian who wishes to increase thinness and / or trim away unwanted fat from pets, the present invention provides means by which this can be achieved. For poultry, cows and pig farmers, using the method of the present invention produces leaner animals that require higher sales prices for the meat industry.

Embodiments of the present invention are illustrated by the following Examples. It should be understood, however, that the embodiments of the invention are not limited to the specific details of these Examples, insofar as other variations thereof will be known or evident in the light of the present disclosure to one skilled in the art.

Examples

Unless otherwise specified, starting materials are generally available from commercial sources, such as Aldrich Chemicals Co. (Milwaukee, Wl), Lancaster Synthesis, Inc. (Windham, NH),

Acros Organics (Fairlawn, NJ), Maybridge Chemical Company, Ltd. (Cornwall, England), Tyger Scientific (Princeton, NJ) and AstraZeneca Pharmaceuticals (London, England).

The acronyms listed below have the following corresponding meanings:

LiN (TMS)₂ - lithium hexamethyldisilazotide

PS-DIEA - polystyrene-linked diisopropylethylamine

AIBN - 2,2'-azo-bis-isobutyronitrile

HOAt -1 -hydroxy-7-azabenzotriazole

EDC - 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

General Experimental Procedures

NMR spectra (NMR) were recorded on a Varian Unity® 400 or 500 (available from Varian Inc., Paio Alto, CA), at room temperature, at 400 and 500 MHz for ¹H, respectively. Chemical shifts are expressed in parts per million (δ) with respect to the residual solvent as an internal reference. Peak shapes are indicated as follows: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br s, broad singlet; v br s, very wide singlet; br m, wide multiplet; 2s, two singles. In some cases, only peak¹Representative H-NMR.

Mass spectra were recorded by direct flow analysis, using positive and negative atmospheric pressure (APcl) chemical ionization examination modes. A Waters APcI / MS ZMD mass spectrometer equipped with a Gilson 215 liquid handling system was used to perform the experiments.

Mass spectrometry analysis was also obtained using the RP-HPLC gradient method for chromatographic separation. The identification of molecular mass was recorded by positive and negative electrospray ionization (ESI) examination modes. A Waters / Micromass ESI / MS model ZMD or LCZ mass spectrometer equipped with Gilson 215 and HP 1100 DAD liquid handling system, was used to carry out the experiments.

Where the intensity of ions containing chlorine or bromine is described, the expected intensity ratio was observed (approximately 3: 1 for ions containing ³⁵CI /³⁷CI and 1: 1 for ions containing ⁷⁹Br /⁸¹Br) and only the lowest mass ion is given. MS peaks are reported for all examples.

Optical rotations were determined on a PerkinElmer® 241 polarimeter (available from PerkinElmer Inc., Wellesley, MA), using sodium line D (λ = 589 nm), at the indicated temperature, and are reported as follows [a]_D^temp, concentration (c = g / 100 mL) and solvent.

Column chromatography was performed either with Baker® silica gel (40 pm; JT Baker, Phillipsburg, NJ) or with Silica Gel 50 (EM Sciences®, Gibbstown, NJ) on glass columns or on Biotage® columns (ISC, Inc., Shelton, CT), under low nitrogen pressure. Radial chromatography was performed using a Chromatotron® (Harrison Research).

Preparation of Key Intermediates

Preparation of Intermediate 1- (2-Chloro-phenyl) -5hydroxy-1 H-pyrazole-3-carboxylic acid ethyl ester (1-1 a)

<img file="BRPI0409701A_D0037.tif" />

To a stirred solution of 2-chlorophenylhydrazine hydrochloride (22.4 g) and potassium carbonate (34.5 g) in ethanol (250 ml) was added diethyl acetylenedicarboxylate (20 ml) and the resulting mixture was heated, reflux for 18 hours. The reaction mixture was cooled and 6 N hydrochloric acid (75 ml) and water (500 ml) were added sequentially. The reaction mixture was extracted with ethyl acetate, the organic layer was washed with water and brine, dried (Na₂ONLY₄) and concentrated in vacuo. The resulting gum was stirred with isopropyl ether (250 ml) to produce the title compound (1-1a). in the form of a brownish solid (23 g), after filtration and drying in vacuo.

Preparation of Intermediate 5-Bromo-1- (2-chloro-55-phenyl) -4-formyl-1 H-pyrazol-3-carboxylic acid ethyl ester (1-1 b)

<img file="BRPI0409701A_D0038.tif" />

To a stirred solution of 1- (2-chlorophenyl) -5-hydroxy-1 H-pyrazol-3-carboxylic acid ethyl ester (1-1 a) (18.2 g) and phosphoryl bromide (39 g) in 1,2-dichloroethane (200 ml) dimethylformamide (10.5 ml) was added over a period of 15 min. The resulting mixture was heated to reflux for 3 hours, cooled, then an additional portion of phosphoryl bromide (98 g) was added, and reflux continued for 20 hours. The black reaction mixture was cooled, poured onto ice (150 g) and stirred for 30 minutes. The mixture was extracted with dichloromethane (2x), the combined organic layers dried over magnesium sulfate and concentrated in vacuo, to produce a dark oil. The oil was passed through a plug of 200 g of silica gel, eluting with 30% hexanes: dichloromethane, to yield the title compound (1-1 b) as a yellow solid, 8.3 g.

Preparation of Intermediate 5-Bromo-1- (2-chlorophenyl) -4- (isopropylamino-methyl) -1 H-pyrazoyl-3-carboxylic acid ethyl ester (1-1c)

<img file="BRPI0409701A_D0039.tif" />

l-1c

To a stirred solution of 5-bromo-1- (2-chloro-phenyl) -4-formyl-1H-pyrazol-3-carboxylic acid ethyl ester (2 g), isopropylamine (0.95 mL) and acetic acid (0 , 4 ml) in 1,2-dichloroethane (16 ml) was added sodium triacetoxy hydroxy borate (1.8 g) and the resulting suspension was stirred for 18 hours. The reaction was diluted with ethyl acetate, washed with saturated aqueous sodium bicarbonate, brine, dried (Na₂ONLY₄) and concentrated in vacuo, to produce the title compound (1-1c), as a golden oil, 2.5 g.

Preparation of Intermediate 3-Bromo-2- (2-chloro-phenyl) -5- (isopropyl-4,5-

<img file="BRPI0409701A_D0040.tif" />

A solution of 5-bromo-1- (2-chloro-phenyl) 4- (isopropylamino-methyl) -1H-pyrazol-3-carboxylic acid ethyl ester (2.2 g) and 1 N aqueous sodium hydroxide ( 33 ml) in ethanol (55 ml) was heated to 50 ° C for 2 hours. The reaction was cooled, acidified to pH ~ 2 with concentrated hydrochloric acid and was concentrated to a vacuum solid. The solids were suspended with ethanol (50 ml), filtered and the filtrate was concentrated in vacuo to produce a white solid, 2.1 g.

To a stirred solution of the above solid (2.0 g), triethylamine (3 ml) in dichloromethane (22 ml) was added 1propanophosphoric acid cyclic anhydride (5 ml of 50% ethyl acetate solution) and the resulting solution was stirred for 20 hours. The reaction was diluted with ethyl acetate, washed with 1 N hydrochloric acid, saturated aqueous sodium bicarbonate, brine and dried (Na₂ONLY₄) to produce the compound mentioned in the title (1 · 1d) as a brownish solid, 2.0 g.

Preparation of Intermediate 5- (4-Chloro-phenyl) -1- (2-chloro-phenyl) -4-1,2- (2-trimethylsilanyl-ethoxy) -vinyl1-1H-pyrazole-3-carboxylic acid ethyl ester (l- 3a)

<img file="BRPI0409701A_D0041.tif" />

A suspension of sodium hydride (45 mg of 60% in oil) in dimethylsulfoxide (2 ml) was stirred at 75 ° C for 45 minutes, cooled to room temperature, then (2- (trimethylsilyl) -ethoxymethyl chloride) triphenylphosphonium (480 mg) was added in one portion to produce a red solution. After 10 minutes, a solution of 5- (4-chloro-phenyl) -1 - (2-chloro-phenyl) -4-formyl-1H-pyrazol-3-carboxylic acid ethyl ester (218 mg) in dimethylsulfoxide (1 ml) was added dropwise and the resulting solution was stirred for 1 hour. Saturated aqueous ammonium chloride was added and the reaction mixture was partitioned between diethyl ether and water. The organic phase was dried (Na₂ONLY₄) and concentrated in vacuo to produce a purple oil. Chromatography on silica gel (30% ethyl acetate / hexanes) gave the title compound (1-3) as a golden oil, 210 mg.

Preparation of Intermediate 5- (4-Chloro-phenyl) -1- (215 chloro-phenyl) -4- (2-oxo-ethyl) -1 H-pyrazol-3-carboxylic acid ethyl ester (l-3b)

<img file="BRPI0409701A_D0042.tif" />

l-3b

A 5- (4-chloro-phenyl) -1- (2-chloro-phenyl) -4- [2- (2-trimethylsilanyl-ethoxy) -vinyl] -1 H-pyrazole-3-carboxylic acid ethyl ester solution l-3a (210 mg) in acetonitrile: conc. (3 ml) was stirred for 2 hours. The reaction was partitioned between saturated aqueous sodium hydrogen carbonate and ethyl acetate. The organic phase was washed with brine, dried (Na₂ONLY₄) and concentrated in vacuo to produce the title compound (1 · 3b), which was taken directly to the next reaction.

Preparation of Intermediate 1- (2-Chloro-phenyl) -5- (4-chloro-phenyl) -4-methyl-1H-pyrazol-3-carboxylic acid ethyl ester (l-4a)

<img file="BRPI0409701A_D0043.tif" />

A solution of 4-chloropropiophenone (16.9 g, 100 mmol) in diethyl ether (20 mL) was added to a LiN solution (TMS)₂ (1 M solution in THF; 100 ml, 100 mmol) in diethyl ether (400 ml), at -78 ° C. The reaction mixture was stirred at -78 ° C for 0.75 h, and then diethyl oxalate (15 ml, 110 mmol) was added dropwise. The reaction mixture was allowed to warm slowly to room temperature and stirred for 17 hours. The diethyl ether was removed in vacuo and the residue was diluted with diethyl ether. A light yellow solid precipitated from the solution and was collected by filtration (9.9 g; 36%). This solid, which was used without further purification, was dissolved in isopropyl alcohol (200 ml) and 2-chlorophenylhydrazine (5.9 g, 36.1 mmol) and H₂ONLY₄ conc. (0.4 mL). The reaction mixture was heated to reflux for 17 hours. After cooling to room temperature, NaHCO was added₃ (1 g). The reaction mixture was concentrated in vacuo. The residue was diluted with EtOAc and the organic solution was washed with NaHCO₃ aq. sat ° and aq. sat, filtered and concentrated in vacuo. The residue was triturated with cyclohexane to give 1-4a, as an off-white solid (9.5 g; 25%): + APCI MS (M + 1) 375.0.

Preparation of Intermediate 4-Bromomethyl-1- (2-chlorophenyl) -5- (4-chloro-phenyl) -1 H-pyrazol-3-carboxylic acid ethyl ester (l-4b)

<img file="BRPI0409701A_D0044.tif" />

A mixture of 1- (2-chloro-phenyl) -5- (4-chloro-phenyl) -4-methyl-1H-pyrazol-3-carboxylic acid ethyl ester l-4a (2.8 G, 7.46 mmol ), Nbromosuccinimide (1.6 g, 8.95 mmol), AIBN (245 mg, 1.49 mmol) in CCI₄(60 ml) was heated to reflux for 17 hours. The reaction was cooled to room temperature, filtered to remove any solids and concentrated in vacuo. The crude residue was purified, via chromatography on silica gel (Flash 40), using a solvent gradient from 10% EtOAc / hexanes to 20% EtOAc / hexanes, to give the desired product (1-4b) as a solid. amorphous (2.2 g; 64%): + APCI MS (M + 1) 455.0. Preparation of Intermediate 1- (2-Chloro-phenyl) -5- (415 chloro-phenyl) -4- (isopropylamino-methyl) -1H-pyrazole-3-carboxylic acid ethyl ester (l-4c)

<img file="BRPI0409701A_D0045.tif" />

A mixture of 4-bromomethyl-1- (2-chlorophenyl) -5- (4-chloro-phenyl) -1H-pyrazol-3-carboxylic acid ethyl ester 1-4b (200 mg, 0.44 mmol); isopropylamine (26 mg, 0.44 mmol), K₂CO₃ (182 mg, 1.32 mmol) in CH₃CN (5 ml) was stirred for 17 hours at room temperature. The reaction mixture was filtered to remove insoluble material and concentrated in vacuo. The residue was diluted with EtOAc and the organic solution was washed with H₂O and saturated aqueous NaCI, dried and concentrated in vacuo. The crude residue was purified via SiO gel chromatography₂, using a solvent gradient from 20% EtOAc / hexanes to 75% EtOAc / hexanes, to 75% EtOAc / hexanes to give the product 1-4c) give the desired product (1-4c) as an amorphous solid (40 mg; 21%): + APCI MS (M + 1) 432.2.

Preparation of Acid Intermediate 1- (2-chloro-phenyl) -5- (4-chloro-phenyl) - 4-

<img file="BRPI0409701A_D0046.tif" />

A solution of 1- (2-chloro-phenyl) -5- (4-chloro-phenyl) -4- (isopropylamino-methyl) -1H-pyrazol-3-carboxylic acid ethyl ester l-4c (32 mg, 0.074 mmol ) in a 1: 4 solution of 1 M KOH / EtOH (10 mL) was stirred at 50 ° C for 6 hours and at 37 ° C for 72 hours. The reaction mixture was treated with conc. until the pH of the solution is approximately 1 and then concentrated in vacuo. The residue was diluted with EtOH and filtered. The filtrate was concentrated in vacuo to give 1-4^The, as a white solid (40 g; 100%): + APCI MS (M + 1) 404.1.

Preparation of Intermediate 2-Chloro-N- (4-chloro-phenyl) -benzamidine (1-5a)

<img file="BRPI0409701A_D0047.tif" />

Triethyl aluminum (2 M in hexanes; 100 mL, 200 mmol) was added dropwise to a solution of 4-chloro-phenylamine (18.2 g, 143 mmol) in toluene (550 mL) under an atmosphere of N₂at 0 ° C. The reaction mixture was warmed to room temperature and stirred for 3.5 hours. A solution of 2-chlorobenzonitrile (23.6 g, 171 mmol) in toluene (140 ml) was added and the reaction mixture was heated at 80 ° C for 17 hours, during which time it became homogeneous. The reaction mixture was then cooled to room temperature and poured over a silica gel slurry in CHCIa / methanol (2: 1). After filtration, the filter cake was washed with a mixture of CFECh / MeOH (2: 1). The combined filtrates were concentrated in vacuo and the solid yellow residue was triturated with hexanes / ether (2: 1). The product l-5a (25.1 g; 66%) was used in the next reaction without further purification.

Preparation of Intermediate 1- (4-Chloro-phenyl) -2- (2-chloro-phenyl) -4- (hydroxy-4.5-dihydro-1H-imidazole-4-carboxylic acid ethyl ester (l-5b)

<img file="BRPI0409701A_D0048.tif" />

A mixture of 2-chloro-N- (4-chloro-phenyl) -benzamidine (1-5a: 25.1 g, 95 mmol) and NaHCO3 (84 g, 189 mmol) in 2-propanol (473 mL) with 3-bromo-2-oxo-propionic acid ethyl ester (14.3 mL, 22 g, 113 mmol). The reaction mixture was heated to 80 ° C for 17 hours. After cooling to room temperature, the solvent was removed in vacuo. The residue was diluted with CH2 Cl2 and the organic solution was washed with H₂O, dried over MgSO4 and concentrated in vacuo, to provide 0- (4-chloro-phenyl) -2- (2-chloro-phenyl) -4- (hydroxy-4,5-dihydro H) acid ethyl ester product -imidazole-4-carboxylic 1-5b, as a dark red residue (36 g).

Preparation of Intermediate 1- (4-Chloro-phenyl) -2- (262 chloro-phenyl) -1 H-imidazole-4-carboxylic acid ethyl ester (l-5c)

<img file="BRPI0409701A_D0049.tif" />

1- (4-Chloro-phenyl) -2- (2-chloro-phenyl) 4- (hydroxy-4,5-dihydro-1 H-imidazole-4-carboxylic acid ethyl ester (1-5b; 36 g, 94.7 mmol) obtained in the previous step and p-toluenesulfonic acid monohydrate (4.5 g, 24 mmol) in toluene (630 mL) was heated under reflux for 17 hours The reaction mixture was cooled to room temperature and concentrated in vacuo. The crude residue was taken up in CH2Cl2 and the organic solution was washed with H₂O, NaHCO₃ aq. sat. and NaCl aq. sat., dried over MgSO₄ and concentrated in vacuo. The crude residue was purified by filtration through a plug of silica gel using a gradient of 2% EtOAc / CH₂CI₂ up to 10% EtOAc / CH₂CI₂. The product-containing fractions were concentrated and the oily residue was diluted with EtOAc / hexanes 1: 3 (200 ml). After 1 hour, a solid precipitated out of solution and was collected by filtration to provide 1- (4-chloro-phenyl) -2- (2-chloro-phenyl) -1H-imidazole-415 carboxylic acid ethyl ester l- 5c (18.63 g; 69.7%).

Preparation of Intermediate 5-Bromo-1- (4-chlorophenyl) -2- (2-chloro-phenyl) -1 H-imidazole-4-carboxylic acid ethyl ester (l-5d)

<img file="BRPI0409701A_D0050.tif" />

Bromine (3.6 mL, 0.07 mol) was added to a 1- (4-chloro-phenyl) -2- (2-chloro-phenyl) -1H-imidazole-4-carboxylic acid ethyl ester solution (1-5c; 3.6 g, 0.01 mol) in glacial acetic acid (50 mL) at room temperature. The reaction mixture was stirred for 17 hours, poured into ice-water and treated with aq. 25% until the orange solution turns yellow. The aqueous solution was extracted with CH₂CI₂ (3x) and the combined extracts were dried and concentrated in vacuo, to give the desired compound 1-5d as an oil (4.7 g): + APCI MS (M + 1) 441.1; ¹H-NMR (CD₃CI) δ 1.41 (3H, t), 4.43 (2H, q), 7.08-7.12 (2H, m), 7.20-7.40 (7H, m).

Preparation of Intermediate 1- (4-Chloro-phenyl) -2- (2-chloro-phenyl) -5-formyl-1H-imidazole-4-carboxylic acid ethyl ester (l-5e)

<img file="BRPI0409701A_D0051.tif" />

To a 5-bromo-1- (4-chlorophenyl) -2- (2-chloro-phenyl) -1H-imidazole-4-carboxylic acid ethyl ester solution (1-5d; 4.4g, 0, 01 mmol) in anhydrous THF (100 mL) and under an atmosphere of N₂ at -78 ° C tert-butyl lithium (13 ml of 1.7 M solution in pentane; 0.22 mol) was slowly added. After 1 hour at -78 ° C, DMF (7.7 mL, 0.1 mmol) was added dropwise. The reaction mixture was stirred at -78 ° C for 2.5 hours, quenched with NH₄CI aq. sat ° (10 mL), allowed to warm slowly to room temperature and finally poured into aq. sat. The aqueous solution was extracted with Et₂The (3x) and the combined organic extracts were dried (Na₂ONLY₄) and concentrated in vacuo. The crude residue was purified via flash chromatography, using a solvent gradient of EtOAc / hexanes 1: 3 to EtOAc / hexanes 1: 1, to give the desired product 1-5e as a pale yellow amorphous vitrified (2.0 g ): + APCI MS (M + 1) 389.2;¹H-NMR (CD₂CI₂) δ 1.41 (3H, t), 4.45 (2H, q), 7.09-7.14 (2H, m), 7.24-7.39 (7H, m), 10.50 ( 1H, s).

The following two intermediates were prepared using procedures analogous to those described above for the synthesis of 1 - (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-formyl-1 H-imidazole- ethyl ester 4-carboxylic (l · 5e):

2- (4-chloro-phenyl) -1- (2-fluoro-phenyl) -5-formyl-1H-imidazole-4-carboxylic acid ethyl ester (1-5e-2);

2- (4-chloro-phenyl) -1- (2-chloro-phenyl) -5-formyl1 H-imidazole-4-carboxylic acid ethyl ester (1-5e-3).

Preparation of Intermediate 2- (4-Chloro-phenyl) -5cyclopentylaminomethyl-1 - (2-fluoro-phenyl) -1 H-imidazole-4-carboxylic acid ethyl ester (l-5f) α

<img file="BRPI0409701A_D0052.tif" />

A 2- (4-chloro-phenyl) -1- (210 fluoro-phenyl) -5-formyl-1H-imidazole-4-carboxylic acid ethyl ester solution l-5e-2 (1000 mg, 2.68 mmol), cyclopentylamine (251 mg, 2.95 mmol), NaBH (OAc)₃ (796 mg, 3.76 mmol) in dichloroethane was stirred at room temperature for 17 h. The reaction was concentrated in vacuo and the residue was diluted with CHCI₃. The organic solution was washed with NaHCO₃ aq. sat. and NaCl aq. sat., dried and concentrated in vacuo. The crude residue was purified on SiO gel₂ (Flash 40s), using a solvent gradient from 30% EtOAc / hexanes to 80% EtOAc / hexanes, to give the desired product (1-5f) as a yellow oil (680 mg; 57%): + APCI MS (M + 1) 442.2.

Preparation of Intermediate 2- (4-Chloro-phenyl) -1- (220 chloro-phenyl) -5- (2-methoxy-vinyl) -1H-imidazole-4-carboxylic acid ethyl ester (l-5q) /

<img file="BRPI0409701A_D0053.tif" />

l-5q

To a solution of (methoxymethyl) triphenylphosphonium chloride (533 mg, 1.55 mmol) in THF (10 mL) at 0 ° C was added lithium hexamethyldisilazotide (1.55 mL, 1 M solution, 1.55 mol). The reaction mixture was stirred for 0.5 h and cooled to -78 ° C. A 25 (4-chloro-phenyl) -1- (2-chloro-phenyl) -5-formyl-1H-imidazole-4-carboxylic acid ethyl ester solution l-5e (408 mg, 1.05 mmol) in THF (5 mL) was added slowly via cannula. The reaction mixture was stirred at -78 ° C for 5 min, then it was allowed to warm to room temperature and stirred for 3 hours. The reaction mixture was quenched with H₂O and diluted with EtOAc. The organic solution was separated and the aqueous layer was extracted with EtOAc. The combined EtOAc extracts were washed with aq. sat., dried and concentrated in vacuo. The crude residue was purified on 4 mm Chromatotron plates using 1: 1 EtOAc / hexanes to give the product (1-5q) as two isomeric compounds (148 mg, 34%, and 157 mg, 36%): + APCI MS (M + 1) 417.2.

Preparation of Intermediate 2- (4-Chloro-phenyl) -1 - {2-chloro-phenyl) -5-f2-oxo-ethyl) -1H-imidazole-4-carboxylic acid ethyl ester (l-5h)

<img file="BRPI0409701A_D0054.tif" />

l-5h

A 2- (4-chloro-phenyl) -1- (2-chloro-phenyl) -5- (2-methoxy-vinyl) -1 H-imidazole-4-carboxylic acid ethyl ester solution (1-5q) ( 275 mg, 0.659 mmol) and H2SO4 (200 μΙ_) in THF / H₂The 5: 1 (18 mL) was heated to 70 ° C for 3 hours. The reaction mixture was cooled to room temperature and treated with K₂CO₃ 1 M until the pH of the reaction mixture is ~ 6. The aqueous solution was extracted with CH₂CI₂ and the combined organic extracts were washed with aq. sat., dried and concentrated in vacuo to give a mixture of the starting material and the product (1-5h): + APCI MS (M + 1) 403.3.

Preparation of Intermediate 2- (4-Chloro-phenyl) -1- (2-chloro-phenyl) -5- (2-cyclopentylamino-ethyl) -1H-imidazole-4-carboxylic acid ethyl ester (Ι-5Π

<img file="BRPI0409701A_D0055.tif" />

P5i

Sodium triacetoxy hydrohydroborate (32 mg, 0.152 mmol) was added to a 2- (4-chloro-phenyl) -1- (2-chlorophenyl) -5- (2-oxo-ethyl) ethyl ester solution -1H-imidazole-4-carboxylic l-5h (34 mg, 0.084 mmol), cy15 clopentylamine (12 pL, 0.118 mmol) and acetic acid (5 pL, 0.09 mmol) in 1,2-dichloroethane (2 mL) at room temperature. The reaction mixture was quenched with 1 N NaOH and extracted with CH₂CI₂ (3x). CH extracts₂CI₂combined were washed with aq. sat, dried and concentrated in vacuo. The crude residue was purified on a 1 mm Chromatotron plate, using 100% EtOAc to give l-5i as a colorless oil (22 mg): + APCI MS (M + 1) 472.2.

Preparation of Intermediate 2- (4-Chloro-phenyl) -1- (2-chloro-phenyl) -5- (2-cyclopentylamino-ethyl) -1 H-imidazole-4-carboxylic acid (l-5i)

ΗΝ

<img file="BRPI0409701A_D0056.tif" />

OH

Cl k5j

To a 2- (4-chloro-phenyl) -1- (2-chloro-phenyl) -5- (2-cyclopentylamino-ethyl) -1H-imidazole-4-carboxylic acid ethyl ester solution (22 mg, 0.46 mol) in absolute EtOH (2 ml) 1 N KOH (500 pL) was added. The reaction mixture was heated to 85 ° C for 4 hours and was then concentrated to a fraction of a quarter of the original volume. The pH of the solution was adjusted to approximately 3.5 using 10% HCI. The aqueous ethanolic solution was concentrated to dryness to give U5j, as a solid (20 mg): + APCI MS (M + 1) 444.4.

Preparation of Intermediate 2- (4-Chloro-phenyl) -5-cyclopentylamino10 methyl) -1 - (2-fluoro-phenyl) -1 H-imidazole-4-carboxylic acid (l-5k) ci l-5k

A 2- (4-chloro-phenyl) -5cyclopentylaminomethyl-1- (2-fluoro-phenyl) -1H-imidazole-4-carboxylic acid ethyl ester solution l-5f (1.2 g, 2.68 mmol ) in 1 M KOH / 1: 2 was stirred at 55 ° C for 17 hours. The reaction mixture was concentrated in vacuo and acidified to pH ~ 1 with concentrated hydrochloric acid. The residue was suspended in EtOH and filtered to remove KCI. The filtrate was concentrated in vacuo, to give 1-5k as an off-white solid (1.26 g, 97%): + APCI MS (M + 1) 414.0.

Example 1 illustrates the preparation of compounds of the present invention where A is nitrogen, B is carbon and X is a bond.

Example 1

Preparation of 2- (2-Chloro-phenyl) -5-isopropyl-3- (3,4.5-trifluoro-phenyl) -4,5-dihydro-2H-pyrrole [3,4-c1pyrazol-6-one (1A- 1)

<img file="BRPI0409701A_D0057.tif" />

1A-1

A solution, purged with nitrogen, of 3-bromo-2- (2-chloro-10-phenyl) -5-isopropyl-4,5-dihydro-2H-pyrrole [3,4-c] pyrazol-6-one l-1d ( 100 mg), cesium fluoride (85 mg), 3,4,5-trifluorophenylboronic acid (74 mg) and tetracis (triphenylphosphine) palladium (0) (32 mg) in 1,2-dimethoxyethane (1 ml) was stirred at a bottle sealed at 80 ° C for 6 hours. The reaction was cooled and partitioned between ethyl acetate / water, the organic phase was dried (Na₂ONLY₄) and concentrated in a vacuum to produce an oil. Phase inversion HPLC (gradient from 40% to 100% acetonitrile: 0.01% aqueous ammonium hydroxide) produced the compound mentioned in the title (1A-1), as a whitish foam, 19 mg.¹H-NMR (CDCI₃) (ppm): δ 7.6-7.4 (m, 4H), 6.78-6.65 (m, 2H), 4.76-4.64 (m, 1H), 4.40 (br s, 2H), 1.36 (d, 6H); ms (LCMS) m / z =

406.3 (M + 1)⁺.

The compounds listed in Table 1 below were prepared using procedures analogous to those described above for the synthesis of Compound 1A-1 using the appropriate commercially available starting materials, prepared using preparations well known to those skilled in the art, or prepared in a manner analogous to the paths described above for other intermediaries.

Table 1

R⁴ i

<img file="BRPI0409701A_D0058.tif" />

<td>Ex.</td><td>R °</td><td>R¹</td><td>R⁴</td><td>LCMS m / z (M + 1)⁺</td>

<td>1A-2</td><td>2-chlorophenyl</td><td>2-chlorophenyl</td><td>-CH (CH₃)₂</td><td> 386,3</td>

<td>1A-3</td><td>2-chlorophenyl</td><td>4- (methoxy-methyl) phenyl</td><td>-CH (CH₃)₂</td><td> 396,4</td>

<td>1A-4</td><td>2-chlorophenyl</td><td>2-fluorophenyl</td><td>-CH (CH₃)₂</td><td> 370,3</td>

<td>1A-5</td><td>2-chlorophenyl</td><td>2-methoxy-pyridyl-5-yl</td><td>-CH (CH₃)₂</td><td> 383,4</td>

<td>1A-6</td><td>2-chlorophenyl</td><td>3-chloro-4-fluorophenyl</td><td>-CH (CH₃)₂</td><td> 404,2</td>

<td>1A-7</td><td>2-chlorophenyl</td><td>4-fluorine-3-methylphenyl</td><td>-CH (CH₃)₂</td><td> 384,3</td>

<td>1A-8</td><td>2-chlorophenyl</td><td>vinyl</td><td>-CH (CH₃)₂</td><td> 302,3</td>

<td>1A-9</td><td>2-chlorophenyl</td><td>4- (trifluoromethyl) -phenyl</td><td>2,2,2-trifluoroethyl</td><td> 460,4</td>

<td>1A-10</td><td>2-chlorophenyl</td><td>4- (trifluoromethyl) -phenyl</td><td>isopropyl</td><td> 420,4</td>

<td>1A-11</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>ethyl</td><td> 372,4</td>

<td>1A-12</td><td>2-chlorophenyl</td><td>2-chlorophenyl</td><td>isopropyl</td><td> 386,3</td>

<td>1A-13</td><td>2-chlorophenyl</td><td>4- (methoxy-methyl) phenyl</td><td>isopropyl</td><td> 396,4</td>

<td>1A-14</td><td>2-chlorophenyl</td><td>2-fluorophenyl</td><td>isopropyl</td><td> 370,3</td>

<td>1A-15</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>2-fluoroethyl</td><td> 390,3</td>

<td>1A-16</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>2,2-difluoroethyl</td><td> 408,1</td>

<td>1A-17</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>2,2,2-trifluoroethyl</td><td> 426,3</td>

<td>1A-18</td><td>2-chlorophenyl</td><td>4-ethoxyphenyl</td><td>f-butifa</td><td> 410,4</td>

<td>1A-19</td><td>2-chlorophenyl</td><td>4-ethoxyphenyl</td><td>/ -butyl</td><td> 410,4</td>

<td>1A-20</td><td>2-chlorophenyl</td><td>4-ethoxyphenyl</td><td>ethyl</td><td> 382,4</td>

<td>1A-21</td><td>2-chlorophenyl</td><td>4-ethoxyphenyl</td><td>isopropyl</td><td> 396,4</td>

<td>Ex.</td><td>R °</td><td>R¹</td><td>R⁴</td><td>LCMS m / z (M + 1)⁺</td>

<td>1A-22</td><td>2-chlorophenyl</td><td>4-ethoxyphenyl</td><td>2,2,2-trifluoroethyl</td><td> 436,4</td>

<td>1A-23</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>f-butyl</td><td> 394,5</td>

<td>1A-24</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>Abutila</td><td> 394,5</td>

<td>1A-25</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>ethyl</td><td> 366,4</td>

<td>1A-26</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>isopropyl</td><td> 380,4</td>

<td>1A-27</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>2,2,2-trifluoroethyl</td><td> 420,4</td>

<td>1A-28</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>2,2-difluoropropyl</td><td> 416,5</td>

<td>1A-29</td><td>2-chlorophenyl</td><td>4-isopropoxyphenyl</td><td>f-butyl</td><td> 424,3</td>

<td>1A-30</td><td>2-chlorophenyl</td><td>4-isopropoxyphenyl</td><td>Abutila</td><td> 424,3</td>

<td>1A-31</td><td>2-chlorophenyl</td><td>4-isopropoxyphenyl</td><td>ethyl</td><td> 396,2</td>

<td>1A-32</td><td>2-chlorophenyl</td><td>4-isopropoxyphenyl</td><td>isopropyl</td><td> 410,2</td>

<td>1A-33</td><td>2-chlorophenyl</td><td>4-isopropoxyphenyl</td><td>2,2,2-trifluoroethyl</td><td> 450,2</td>

<td>1A-34</td><td>2-chlorophenyl</td><td>4-isopropoxyphenyl</td><td>2,2-difluoropropyl</td><td> 446,3</td>

<td>1A-35</td><td>2-chlorophenyl</td><td>4-f-butylphenyl</td><td>2,2,2-trifluoroethyl</td><td> 448,5</td>

<td>1A-36</td><td>2-chlorophenyl</td><td>4-f-butylphenyl</td><td>2,2-difluoropropyl</td><td> 444,5</td>

<td>1A-37</td><td>2-chlorophenyl</td><td>4-i-propylphenyl</td><td>2,2,2-trifluoroethyl</td><td> 434,5</td>

<td>1A-38</td><td>2-chlorophenyl</td><td>4-i-propylphenyl</td><td>2,2-difluoropropyl</td><td> 430,5</td>

Preparation of 2- (2-Chloro-phenyl) -3- [2- (4-chloro-phenyl) -vinin-5-isopropyl-4,5dihydro-2H-pyrroir3,4-c1pyrazol-6-one (1 A- 39)

<img file="BRPI0409701A_D0059.tif" />

1A-39

To a stirred solution of 2- (2-chloro-phenyl) -5-isopropyl-3-vinyl4,5-dihydro-2H-pyrrole [3,4-c] pyrazol-6-one 1A-39 (42 mg), palladium acetate (3 mg) and 4-chloroiodobenzene (300 mg) were stirred for 18 hours. The reaction was concentrated and chromatographed on silica gel (30% to 60% ethyl acetate gradient / hexanes) to produce the title compound (1A-39). 44 mg.¹H-NMR (d₆-DMSO) (ppm): δ 7.78 (d, 1H), 7.63-7.38 (m, 5H), 7.13 (d, 1H), 6.61 (d, 1H), 4, 62 (s, 2H), 4.40 (m, 1H), 1.23 (d, 6H); ms (LCMS) m / z = 412.3 (M + 1)⁺.

Example 2 illustrates the preparation of compounds of the present invention where A is nitrogen, B is carbon and X is -C (R^2a) (R^2b)-.

Example 2

Preparation of 3- (4-Chloro-phenyl) -2- (2-chloro-phenyl) -6-isopropyl-2,4,5,6-tetrahydropyrazoloí3.4-c1pyridin-7-one (2A-1)

<img file="BRPI0409701A_D0060.tif" />

To a stirred solution of l-3b (157 mg), isopropylamine (66 pL), 15 acetic acid (27 μΙ_) in 1,2-dichloroethane (0.5 ml) was added sodium triacethoxyhydro-borate (124 mg). After 1.5 hours, the reaction was diluted with ethyl acetate, washed with saturated aqueous sodium bicarbonate, brine, dry (Na₂ONLY₄) and concentrated in vacuo, to produce a golden foam, which was taken to the next step without further purification.

The product from the above step and 1N aqueous sodium hydroxide (2.5 ml) were heated to 50 ° C for 2.5 hours. The reaction solution was cooled, acidified to pH ~ 2 with concentrated hydrochloric acid and concentrated in vacuo. The resulting solid residue was suspended in ethanol (10 ml), filtered, the solids were washed with ethanol and the combined filtrates were concentrated in vacuo, to produce a white solid, which was taken to the next step without further purification.

To a stirred solution of the solid prepared above in the previous step, triethylamine (0.2 ml) in dichloromethane (3 ml) were added to 1-propanophosphoric acid cyclic anhydride (0.34 ml of 50% ethyl acetate solution) and the resulting solution was stirred for 2 hours. The reaction mixture was diluted with ethyl acetate, washed with 1 N hydrochloric acid, saturated aqueous sodium bicarbonate, brine and dried (Na₂SO4) to produce a golden oil. Chromatography on silica gel (60% ethyl acetate / hexanes) gave the title compound (2A-1) as a white solid, 103 mg.¹H-NMR (CDCI₃) (ppm): δ 7.50-7.23 (m, 6H), 7.03 (d, 2H), 5.17 (m, 1H), 3.57 (m, 2H), 2.84 ( br s, 2H), 1.20 (d, 6H); ms (LCMS) m / z = 400.3 (M + 1)⁺.

The compounds listed in Table 2 below were prepared using procedures analogous to those described above for the synthesis of Compound 2A-1. using the appropriate starting materials that are commercially available, prepared using preparations well known to those skilled in the art, or prepared in a manner analogous to the routes described above for other intermediates.

Table 2

<img file="BRPI0409701A_D0061.tif" />

<td>Ex.</td><td>R °</td><td>R¹</td><td>R⁴</td><td>LCMS m / z (M + go</td>

<td>2A-2</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>2,2,2-trifluoroethyl</td><td> 440,2</td>

<td>2A-3</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>2,2-difluoroethyl</td><td> 422,3</td>

<td>2A-4</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>2-fluoroethyl</td><td> 404,3</td>

<td>Ex.</td><td>R °</td><td>R¹</td><td>R⁴</td><td>LCMS m / z (M + 1)⁺</td>

<td>2A-5</td><td>2-chlorophenyl</td><td>4- (trifluoromethyl) - phenyl</td><td>isopropyl</td><td> 434,4</td>

<td>2A-6</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>ethyl</td><td> 386,4</td>

<td>2A-7</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>X)</td><td> 426,4</td>

<td>2A-8</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td></td><td> 412,4</td>

<td>2A-9</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>f-butyl</td><td> 414,4</td>

<td>2A-10</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>/ -butyl</td><td> 414,4</td>

<td>2A-11</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>2,2-difluoropropyl</td><td> 436,3</td>

<td>2A-12</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>'X</td><td> 454,5</td>

<td>2A-13</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>2-methoxy-2- methylpropyl</td><td> 444,4</td>

<td>2A-14</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>2,2-difluoropropyl</td><td> 430,5</td>

<td>2A-15</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>isopropyl</td><td> 394,5</td>

<td>2A-16</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>i-butyl</td><td> 408,5</td>

<td>2A-17</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>/ -butyl</td><td> 408,4</td>

<td>2A-18</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>2,2,2-trifluoroethyl</td><td> 434,4</td>

<td>2A-19</td><td>2-chlorophenyl</td><td>4-ethylphenyl</td><td>ethyl</td><td> 380,4</td>

<td>2A-20</td><td>2-chlorophenyl</td><td>4-isopropylphenyl</td><td>2,2-difluoropropyl</td><td> 444,5</td>

<td>2A-21</td><td>2-chlorophenyl</td><td>4-isopropylphenyl</td><td>2,2,2-trifluoroethyl</td><td> 448,5</td>

<td>2A-22</td><td>2-methylphenyl</td><td>4-chlorophenyl</td><td>2,2,2-trifluoroethyl</td><td> 420,4</td>

<td>2A-23</td><td>2-methylphenyl</td><td>4-chlorophenyl</td><td>2,2-difluoropropyl</td><td> 416,4</td>

Example 3

Preparation of 3- (4-Chloro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl4,5-dihydro-2H-pyrrolf3,4-c1pyrazol-6-one (3A-1)

<img file="BRPI0409701A_D0062.tif" />

A mixture of 1- (2-chloro-phenyl) -5- (4-chloro-phenyl) -4- (isopropylamino-methyl) -1H-pyrazol-3-carboxylic acid l-4d (40 mg, 0.074 mmol), EDC (28 mg, 0.148 mmol), HOAt (20 mg, 0.148 mmol) and triethylamine (0.02 mL, 0.148 mmol) in CH2 Cl2 (10 mL) was stirred at room temperature for 17 hours. The reaction mixture was washed with NaHCO₃ aq. sat., NaCl aq. sat., dried and concentrated in vacuo. The crude residue was diluted with cyclohexane and stirred for 17 hours. A solid precipitated out of solution and was collected by filtration to give 3A-1 (11 mg; 38%): + APCI MS (M + 1) 386.1;¹H-NMR (CDCI₃) δ 7.50-7.38 (m, 4H), 7.26 (d, 2H, J = 9.6 Hz), 7.06 (d, 2H), 4.69 (m,

1H), 1.3 (d, 6H, J = 6.65 Hz).

The compounds listed in Table 3 below were prepared using procedures analogous to those described above for the synthesis of Compound 3A-1 using the appropriate commercially available starting materials, prepared using preparations well known to those skilled in the art, or prepared in a similar manner. to the paths described above for other intermediaries.

<img file="BRPI0409701A_D0063.tif" />

O

<td>Ex.</td><td>R °</td><td>R¹</td><td>R³³</td><td>RSh</td><td>R⁴</td><td>+ APCI MS (M + 1)</td>

<td>3A-2</td><td>2-chlorophenyl</td><td>4-methoxyphenyl</td><td>H</td><td>H</td><td>x&gt;</td><td> 408,2</td>

<td>3A-3</td><td>2-chlorophenyl</td><td>4-methoxyphenyl</td><td>H</td><td>H</td><td>X)</td><td> 422,4</td>

<td>3A-4</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>H</td><td>H</td><td>Λ</td><td> 400,0</td>

<td>3A-5</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>H</td><td>H</td><td>Λ /</td><td> 398,2</td>

<td>3A-6</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>H</td><td>H</td><td></td><td> 412,3</td>

<td>3A-7</td><td>2,4-dichlorophenyl</td><td>4-chlorophenyl</td><td>H</td><td>H</td><td>Λ</td><td> 420,0</td>

<td>3A-8</td><td>2,4-dichlorophenyl</td><td>4-chlorophenyl</td><td>H</td><td>H</td><td>Λ</td><td> 434,1</td>

<td>3A-9</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td>Λ</td><td> 386,1</td>

<td>3A-10</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td></td><td> 412,4</td>

<td>3A-11</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td>X)</td><td> 426,4</td>

<td>3A-12</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td></td><td> 438,1</td>

<td>3A-13</td><td>4-chlorophenyl</td><td>2-fluorophenyl</td><td>H</td><td>H</td><td></td><td> 396,4</td>

<td>3A-14</td><td>4-chlorophenyl</td><td>2-fluorophenyl</td><td>H</td><td>H</td><td>X</td><td> 410,5</td>

<td>3A-15</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td>Xr</td><td> 427,3</td>

<td>3A-16</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>H</td><td>H</td><td></td><td>427.3 (M + 1 -Boc)</td>

<td>3A-17</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td></td><td> 527,1</td>

<td>3A-18</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td>^ j ~ Z /^nboc</td><td> 513,1</td>

Example 4 illustrates the preparation of compounds of the present invention where A is carbon, B is nitrogen (imidazole derivatives), and X is a bond.

Example 4

Preparation of 2- (4-Chloro-phenyl) -5-cyclopentyl-1 - (2-fluoro-phenyl) -5,6-dihydro-1 Hpirrolf3,4-d1imidazole-4-one (4A-1) «·

<img file="BRPI0409701A_D0064.tif" />

α

4Α-1

A mixture of 2- (4-chloro-phenyl) -5-cyclopentylaminomethyl-1- (2-fluoro-phenyl) -1H-imidazole-4-carboxylic acid l-5k (1.2 g, 2.59 mmoles), EDC (994 mg, 5.18 mmoles), HOAt (704 mg, 5.18 mmoles) and triethylamine (1.1 ml, 7.76 mmoles) in CH2 Cl2 (200 ml) was stirred at room temperature for 17 hours. The reaction mixture was washed with NaHCO₃ aq. sat., NaCl aq. sat, dried and concentrated in vacuo. The crude residue was purified on SiO gel₂ using a solvent gradient from 30% EtOAc / hexanes to 60% EtOAc / hexanes, to give 4A-1 as a white solid (752 mg; 73%): + APCI MS (M + 1) 396.2; ¹H-NMR (CDCI₃) Ô 7.61-7.59 (m, 2H), 7.42-7.37 (m, 6H), 4.60 (m, 1H), 4.50 (s, 2H), 2.0 ( m, 2H), 1.8-1.65 (m, 6H).

The compounds listed in Table 4 below were prepared using procedures analogous to those described above for the synthesis of Compound 4A-1, using the appropriate commercially available starting materials, prepared using preparations well known to those skilled in the art, or prepared in a manner analogous to the paths described above for other intermediaries. The compounds listed below were initially isolated as their free base and then converted to their corresponding hydrochloride salt before testing in vivo (if tested in vivo).

Table 4

<img file="BRPI0409701A_D0065.tif" />

O

<td>Ex.</td><td>R °</td><td>R¹</td><td>R</td><td>+ ES MS (M + 1)</td>

<td>4A-2</td><td>4-chlorophenyl</td><td>2,4-dichlorophenyl</td><td></td><td> 446,0</td>

<td>4A-3</td><td>4-chlorophenyl</td><td>2,4-dichlorophenyl</td><td>X)</td><td> 460,0</td>

<td>4A-4</td><td>4-chlorophenyl</td><td>2,4-dichlorophenyl</td><td>-ch₂cf₃</td><td> 460,0</td>

<td>4A-5</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>Λ</td><td> 386,3</td>

<td>4A-6</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td></td><td> 412,2</td>

<td>4A-7</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>X</td><td> 426,0</td>

<td>4A-8</td><td>4-chlorophenyl</td><td>2-fluorophenyl</td><td>X)</td><td> 410,2</td>

<td>4A-9</td><td>4-chlorophenyl</td><td>2-fluorophenyl</td><td>XJ</td><td> 424,3</td>

<td>4A-10</td><td>4-fluorophenyl</td><td>2-fluorophenyl</td><td>xO</td><td> 380,2</td>

<td>4A-11</td><td>4-fluorophenyl</td><td>2-chlorophenyl</td><td>xO</td><td> 396,2</td>

<td>4A-12</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>xO</td><td> 412,2</td>

<td>4A-13</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>X</td><td> 426,2</td>

<td>4A-14</td><td>2,4-dichlorophenyl</td><td>4-chlorophenyl</td><td></td><td> 460,2</td>

Example 5 illustrates the preparation of compounds of the present invention where A is carbon, B is nitrogen and X is -C (R^2a) (R^2b)-.

Example 5

Preparation of 2- (4-Chloro-phenyl) -1 - (2-chloro-phenyl) -5-cyclopentyl-1,5,6,7-tetrahi5 dro-imidazoí4.5-clpiridin-4-one (5A-1 )

<img file="BRPI0409701A_D0066.tif" />

A mixture of 2- (4-chloro-phenyl) -1- (2-chloro-phenyl) -5- (2-cyclopentylamino-ethyl) -1H-imidazole-4-carboxylic acid l-5i (20 mg, 0.046 mmol ), EDC (19mg, 0.1mmol), HOAt (13mg, 0.1mmol) and triethylamine (14 μΐ_, 0.1mmol) in 1,2-dichloroethane (20ml) was stirred at room temperature, for 17 hours. The reaction mixture was washed with NaHCO₃ aq. sat ° and the aqueous bicarbonate solution was extracted again with CH2 Cl2. The combined CH2 Cl2 extracts were dried and concentrated in vacuo. The residue was dissolved in diethyl ether (1 ml) and several drops of 4 M HCI in dioxane were added. The organic solution was decanted and more ether was added.

The mixture was stirred for several minutes and the solvent was decanted again. The residue was dried in vacuo to give the desired product 5A-1. as a colorless solid: + APCI MS (M + 1) 426.3;¹H-NMR (CDCI₃) δ 7,627.57 (m, 1H), 7.53-7.47 (m, 1H), 7.44-7.39 (m, 1H), 7.37-7.33 (m, 2H), 7.327.28 (m, 1H), 7.24-7.19 (m, 2H), 5.16-5.06 (m, 1H), 3.56-3.50 (m, 2H), 3, 2615 3.17 (m, 2H), 2.71-2.56 (m, 2H), 1.92-1.83 (m, 2H), 1.76-1.46 (m, 4H).

Example 6 illustrates the preparation of compounds of the present invention where A is nitrogen, B is carbon, X is a bond and R^3rd and R^3b are hydrogen, alkyl and arylalkyl.

Example 6

Preparation of 3- (4-Chloro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl-4-methyl-4,5-dihydro-2H-pyrroir3,4-clpyazol-6-one (6AA)

V

<img file="BRPI0409701A_D0067.tif" />

To a solution of 3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl4,5-dihydro-2H-pyrrol [3,4-c] pyrazol-6-one 3A-1 (19 mg, 0.05 mmol) in THF (0.5 mL) at -78 ° C LiHMDSi (55 µL, 0.055 mmol) was added. A very dark blue solution was formed. The reaction mixture was stirred for 0.17 hours and then iodomethane (4.4 µL, 0.07 mmol) was added dropwise. The reaction mixture was stirred at -78 ° C for 0.25 h (until a yellow color was formed) and at room temperature for 2 hours, quenched with NH₄CI aq. sat. and extracted with EtOAc. The organic solution was washed with aq. sat., dried and concentrated in vacuo. The residue was purified on 1 mm Chromatotron plates using a 1: 1 solution of EtOAc / hexane to give 6A-1 as a white solid (3.2 mg; 14%); + ES MS (M + 1) 400.3;¹H-NMR (CDCI₃): δ 7.4-7.3 (m, 4H), 7.28-7.24 (m, 2H), 7.1-7.06 (m, 2H), 4.84-4.77 ( m, 1H), 4.37-4.27 (m, 1H), 1.48-1.4 (m, 9H).

The compounds listed in Table 6 below were prepared using procedures analogous to those described above for the synthesis of Compound 6A-1, using the appropriate commercially available starting materials, prepared using preparations well known to those skilled in the art, or prepared in a manner analogous to the paths described above for other intermediaries.

Table 6

<img file="BRPI0409701A_D0068.tif" />

O

<td>Ex.</td><td>R °</td><td>R¹</td><td>R^3rd</td><td>R^3b</td><td>R⁴</td><td>+ ES MS (M + H)</td>

<td>6A-2</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>X</td><td>H</td><td>Λ</td><td> 476,3</td>

<td>6A-3</td><td>2-chlorophenyl</td><td>4-chlorophenyl</td><td>X</td><td>X</td><td>Λ</td><td> 566,4</td>

Example 7 illustrates the preparation of compounds of the present invention where A is nitrogen, B is carbon, X is a bond and R⁴ is pyrrolidin3-yl, piperidin-3-yl, piperidin-4-yl.

Example 7

Preparation of 3- (4-Chloro-phenyl) -2- (2-chloro-phenyl) -5- (1-isopropyl-piperidin-4-yl) 4,5-dihydro-2H-pyrroir3.4-c1pyrazol-6 -one (7A-1)

<img file="BRPI0409701A_D0069.tif" />

A solution of 4- [3- (4-chloro-phenyl) 2- (2-chloro-phenyl) -6-oxo-2,6-dihydro-4H-pyrrole [3,4- tert-butyl ester] c] pyrazol-5-yl] -piperidine-1-carboxylic acid 3A-17 (80 mg, 0.152 mmol) in conc. HCI / EtOH 1: 5 (6 mL) was stirred at room temperature for 2 hours. The reaction mixture was concentrated in vacuo, to give a white solid.

A mixture of the product obtained in the previous step (30 mg, 0.065 mmol), 2-bromopropane (24 mg, 0.194 mmol); K2CO3 (45 mg, 0.323 mmol) in DMF (2 mL) was stirred at room temperature for 17 hours. The reaction mixture was diluted with EtOAc and washed with H₂O and NaCI aq. sat, dried and concentrated in vacuo. The crude residue was azeotroped once with heptane, to remove any DMF, and purified via SiO gel chromatography₂, using Et₂5% NH / EtOAc to give an oil. ¹H-NMR (CDCb): δ 7.62-7.45 (m, 4H), 7.39-7.32 (d, 2H), 7.32-7.2 (d, 2H), 4.60 (s,

2H), 4.18 (m, 1H), 3.10-3.03 (m, 1H), 1.96-1.90 (m, 4H), 1.30-1.25 (m, 4H) ,

1.09-1.07 (d, 6H).

The product of the above reaction was stirred in 4 M HCI / dioxane (1 ml) for 0.25 hours, and concentrated in vacuo to give 7A-1 as an amorphous solid (2 mg; 6%).

The compounds listed in Table 7 below were prepared using procedures analogous to those described above for the synthesis of Compound 7A-1, using the appropriate commercially available starting materials, prepared using preparations well known to those skilled in the art, or prepared in a manner analogous to the paths described above for other intermediaries. The compounds listed below were generally isolated as the free base and then converted to their corresponding hydrochloride salts, before testing in vivo (if tested in vivo).

Table 7

<img file="BRPI0409701A_D0070.tif" />

i

<td>Ex.</td><td>R °</td><td>R¹</td><td>R^3rd</td><td>R^3b</td><td>R⁴</td><td>+ ESMS (M + H)</td>

<td>7A-2</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td></td><td> 469,2</td>

<td>7A-3</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td></td><td> 455,2</td>

<td>7A-4</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td></td><td> 469,1</td>

<td>7A-5</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td>XX,</td><td> 455,1</td>

<td>7A-6</td><td>4-chlorophenyl</td><td>2-chlorophenyl</td><td>H</td><td>H</td><td></td><td> 455,3</td>

Example 8 illustrates the preparation of compounds of the present invention having Formula (II) or (IV).

Example 8

Preparation of 3- (4-Chloro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl-2,4,5.6-tetrahydro82 pyrrole [3.4-clpyazole (8A-1)

<img file="BRPI0409701A_D0071.tif" />

A solution of 3- (4-chloro-phenyl) -2- (2-chloro-phenyl) -5-isopropyl-4,5dihydro-2H-pyrrol [3,4-c] pyrazol-6-one 3A-1 ( 7 mg, 0.018 mmol) and BH₃THF (166 mL, 166 mmol) was stirred at room temperature for 1 hour and at 50 ° C for 17 hours. After the reaction mixture had cooled to room temperature, MeOH (5 ml) was added. The reaction mixture was heated under reflux for 2 h, cooled to room temperature, and concentrated in vacuo. The residue was diluted with 4 M HCI / dioxanes (1 ml) and concentrated in vacuo. The residue was dissolved in CH₂CI₂ and he10 xanes were added to precipitate 8A-1. as a colorless solid (2 mg; 27%): + APCI MS (M + 1) 372.5;¹H-NMR (CD₃OD): δ 7.59-7.43 (m, 4H), 7.38 (d, 2H), 7.20 (d, 2H), 4.80-4.65 (m, 2H), 3, 91-3.82 (m, 1H), 3.68-3.55 (m, 2H), 1.52 (d, 6H).

The compounds listed in Table 8 below were prepared using procedures analogous to those described above for the synthesis of Compound 8A-1. using the appropriate starting materials that are commercially available, prepared using preparations well known to those skilled in the art, or prepared in a manner analogous to the routes described above for other intermediates. The compounds listed below were generally isolated as the free base and then converted to their corresponding hydrochloride salts prior to in vivo testing (if tested in vivo).

Table 8

R⁴ ι

<img file="BRPI0409701A_D0072.tif" />

<td>Ex. No.</td><td>R °</td><td>R¹</td><td>R⁴</td><td>MS (MH)⁺</td>

<td>8A-2</td><td>4-chlorophenyl</td><td>2-fluorophenyl</td><td>Ni</td><td> 396,5</td>

<td>8A-3</td><td>2,4-dichlorophenyl</td><td>4-chlorophenyl</td><td></td><td> 434,4</td>

Pharmacological Tests

The usefulness of the compounds of the present invention in the practice of the present invention, can be evidenced by the activity in at least one of the protocols described here below. The following acronyms are used in the protocols described below.

BSA - bovine serum albumin

DMSO - dimethylsulfoxide

EDTA - ethylenediaminetetraacetic acid

PBS - phosphate buffered saline

EGTA - ethylene glycol-bis (p-aminoethyl ether) -N, N, N ', N'tetraacetic acid

GDP - guanosine-diphosphate sc - subcutaneous po - orally ip - intraperitoneal icv - intracerebro-ventricular iv - intravenous [³H] SR141716A - N- (piperidin-1-yl) -5- (4-chlorophenyl) -1- (2,4dichlorophenyl) -4-methyl-1H-pyrazol-3-carboxamide hydrochloride, available in Amersham Biosciences, Piscataway, NJ.

[³H] CP-55940 - 5- (1,1-dimethylheptyl) -2- [5-hydroxy-2- (384 hydroxypropyl) -cyclohexyl] -phenol radioactively available from NEN Life Science Products, Boston, MA .

AM251 -A / - (piperidin-1-yl) -1 - (2,4-dichlorophenyl) -5- (4-iodophenyl) -4methyl-1H-pyrazol-3-carboxamide available fromTocris ™, Ellisville, MO.

All compounds listed in the Examples section above were tested in the CB-1 receptor binding assay below. The compounds provided a range of binding activities in the range 0.6 nM - 2500 nM.

Those compounds that have an activity &lt;20 nM were then tested in the CB-1 Binding Assay ΘΤΡ_γ [³⁵S] and in the CB-2 binding assay described below in the section Biological Binding Assays. The selected compounds were then tested in vivo using one or more of the functional assays described in the Functional Biological Assays section below.

In Vitro Biological Assays

Bioassay systems for determining CB-1 and CB-2 binding properties and cannabinoid receptor ligand pharmacological activity are described by Roger G. Pertwee in Pharmacology of Cannabinoid Receptor Ligands in Current Medicinal Chemistry, 6 (1999) 635-664, and WO 92/02640 (US Application No. 07/564 075, filed August 8, 1990, incorporated herein by reference).

The following assays are designed to detect compounds that inhibit [³H] SR141716A (radioactive selective labeled CB-1 ligand) and [³H] 5- (1,1-dimethylheptyl) -2- [5-hydroxy-2- (3-hydroxypropyl) cyclohexyl-phenol; radiolabeled CB-1 / CB-2 ligand) to their respective receptors.

Mouse CB-1 Receiver Binding Protocol

PelFreeze brains (available from Pel Freeze Biologicals, Rogers, Arkansas) were cut and placed in tissue preparation buffer (Tris 5 mM-HCI, pH = 7.4, and 2 mM EDTA), politronated at high speed and kept on ice for 15 minutes. The homogenate was then vortexed at 1000 xg for 5 minutes at 4 ° C. The supernatant was recovered and centrifuged at 100,000 xg for 1 hour at 4 ° C. The pellet was then resuspended in 25 mL of TME (25 nM Tris, pH = 7.4, MgCl₂ 5 mM and 1 mM EDTA), per brain used. A protein assay was performed and 200 pL of tissue, totaling 20 pg, was added to the assay.

The test compounds were diluted in drug buffer (0.5% BSA; 10% DMSO and TME) and then 25 µl was added to a deep cavity polypropylene plate. [³H] SR141716A was diluted in a binder buffer (0.5% BSA plus TME) and 25 µl was added to the plate. A BCA protein assay was used to determine the appropriate tissue concentration and then 200 µL of rat brain tissue, at the appropriate concentration, was added to the plate. The plates were covered and placed in an incubator at 20 ° C for 60 minutes. At the end of the incubation period, 250 pL of final buffer (5% BSA plus TME) was added to the reaction plate. The plates were then harvested by Skatron on GF / B filtration materials pre-soaked in BSA (5 mg / mL) plus TME. Each filter was washed twice. The filters were dried overnight. In the morning, the filters were counted in a Wallac Betaplate ™ counter (available from PerkinElmer Life Sciences ™, Boston, MA).

Human CB-1 Receiver Binding Protocol

Human embryonic kidney cells 293 (HEK 293), 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, bicarbonate 10 mM Na, protease inhibitors; pH = 7.4) and homogenized with a Dounce homogenizer. The homogenate was then vortexed at 1000 xg for 5 minutes at 4 ° C. The supernatant was recovered and centrifuged at 25,000 xg for 20 minutes at 4 ° C. The pellet was then resuspended in 10 ml of homogenization buffer and again stirred at 25,000 xg for 20 minutes at 4 ° C. The final pellet was resuspended in 1 mL of TME (25 mM Tris buffer (pH = 7.4) containing MgCI₂ 5 mM and 1 mM EDTA). A protein test was performed and 200 μΙ_ of tissue, totaling 20 pg, was added to the test.

The test compounds were diluted in drug buffer (0.5% BSA; 10% DMSO and TME) and then 25 μΙ_ was added to a deep cavity polypropylene plate. [³H] SR141716A was diluted in a binder buffer (0.5% BSA plus TME) and 25 μ! _ Was added to the plate. The plates were covered and placed in an incumbent at 30 ° C for 60 minutes. At the end of the incubation period, 250 μΙ_ of final buffer (5% BSA plus TME) was added to the reaction plate. The plates were then harvested by Skatron on GF / B filtration materials pre-soaked in BSA (5 mg / mL) plus TME. Each filter was washed twice. The filters were dried overnight. In the morning, the filters were counted in a Wallac Betaplate ™ counter (available from PerkinElmer Life Sciences ™, Boston, MA).

CB-2 Receiver Connection Protocol

Chinese Hamster K1 ovary cells (CHO-K1), transfected with CB-2 cDNA (obtained from Dr. Debra Kendall, University of Connecticut), were harvested in tissue preparation buffer (Tris 5 mM-HCI ( pH = 7.4) containing 2 mM EDTA), politronated at high speed and kept on ice for 15 minutes. The homogenate was then vortexed at 1000 xg for 5 minutes at 4 ° C. The supernatant was recovered and centrifuged at 100,000 xg for 1 hour at 4 ° C. The pellet was then resuspended in 25 mL of TME (25 mM Tris buffer (pH = 7.4) containing MgCI₂ 5 mM and 1 mM EDTA) per brain used. A protein assay was performed and 200 pL of tissue totaling 10 pg was added to the assay.

The test compounds were diluted in drug buffer (0.5% BSA; 10% DMSO and 80.5% TME) and then 25 µl was added to a deep cavity polypropylene plate. [³H] 5- (1,1-dimethylheptyl) -2- [5hydroxy-2- (3-hydroxypropyl) -cyclohexyl] -phenol was diluted in a binder buffer (0.5% BSA and 99.5% TME ) and then 25 pL 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 μΙ of the tissue at the appropriate concentration was added to the plate. The plates were covered and placed in an incumbent at 30 ° C for 60 minutes. At the end of the incubation period, 250 μΙ_ of final buffer (5% BSA plus TME) was added to the reaction plate. The plates were then harvested by Skatron on GF / B filtration materials pre-soaked in BSA (5 mg / mL) plus TME. Each filter was washed twice. The filters were dried overnight. The filters were then counted in a Wallac Betaplate ™ counter.

GTP Binding Assay_y f³⁵Sl to CB-1

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 in Molecular Pharmacology, 50 (1996) 709-715. GTP binding assays_Y[³⁵S] were performed in 96-well FlashPlate ™ format, in duplicate, using GTPy [³⁵S] 100 pM and 10 pg of membrane per well, in assay buffer composed of 50 mM Tris-HCI, pH 7.4, 3 mM MgCl2, pH 7.4, 10 mM MgCl2, 20 mM EGTA, 100 mM NaCI, GDP 30 μΜ, 0.1% bovine serum albumin and the following protease inhibitors: bacitracin 100 pg / mL, benzamidine 100 pg / mL, aprotinin 5 pg / mL, leupeptin 5 pg / mL. The test mixture was then incubated with increasing concentrations of antagonist (10 '^1θ M to 10 '⁵ M), for 10 minutes, stimulated with the cannabinoid agonist 5- (1,1-dimethylheptyl) -2- [5-hydroxy-2- (3-hydroxypropyl) cyclohexyl-phenol (10 pM). The tests were carried out at 30 ° C, for one hour. The FlashPlates ™ were then centrifuged at 2000 xg for 10 minutes. Stimulation of GTPy binding [³⁵S] was then quantified using a Wallac Microbeta. ED50 calculations made using Prism ™ by Graphpad.

The inverse agonism was measured in the absence of an agonist. Functional Assay Protocol Based on CB-1 FLIPR

CHO-K1 cells co-transfected with the human CB-1 receptor cDNA (obtained from Dr. Debra Kendall, University of Connecticut) and the promiscuous G G16 protein were used for this assay. The cells were plated 48 hours before, at 12,500 cells per well, in clear, collagen-coated 384-well black assay plates. The cells were incubated for one hour with Fluo-4 AM 4 μΜ (Molecular Probes) in DMEM (Gibco) containing 2.5 mM probenicide and pluronic acid (0.04%). The plates were then washed 3 times with HEPES buffered saline (containing probenicide; 2.5 mM), to remove excess dye. After 20 minutes, the plates were added to FLIPR individually and the fluorescence levels were continuously monitored over a period of 80 seconds. Compound additions were made simultaneously to all 384 wells, 20 seconds after the baseline. The tests were performed in triplicate and 6 points of the concentration response curve were generated. The antagonist compounds were subsequently stimulated with WIN 55 212-2 (agonist) 3 μΜ. The data were analyzed using Graph Pad Prism.

Detection of Inverse Agonists

The following cyclic AMP assay protocol using intact cells was used to determine inverse agonist activity.

The cells were plated on a 96-well plate, with a density of 10,000 to 14,000 cells per well, at a concentration of 100 μΙ_ per well. The plates were incubated for 24 hours in an incubator at 37 ° C. The medium was removed and serum (100 μΙ_) lacking medium was added. The plates were then incubated for 18 hours at 37 ° C.

Serum-free medium, containing 1 mM IBMX, was added to each well, followed by 10 pL of test compound (stock solution (25 mM compound in DMSO) in 50% DMSO / PBS 1:10) diluted 10x in PBS with 0.1% BSA). After incubation for 20 minutes at 37 ° C, forescoline 2 μΜ was added and then incubated for another 20 minutes at 37 ° C. The medium was removed, 100 μΐ of 0.01 N HCI was added and then incubated for 20 minutes at room temperature. The cells lysed (75 μΙ_) along with 25 μΙ_ of assay buffer (supplied in a cAMP FtashPlate ™ assay kit, available from NEN Life Science Products, Boston, MA), in a FlashPlate. CAMP and cAMP tracer standards were added, following the kit protocol. The FlashPlate was then incubated for 18 hours at 4 ° C. The contents of the wells were aspirated and counted in a scintillation counter.

In Vivo Biological Assays

Cannabinoid agonists such as A⁹-tetrahydrocannabinol (Δ⁹THC) and 5- (1,1-dimethylheptyl) -2- [5-hydroxy-2- (3-hydroxypropyl) -cyclohexyl] -phenol have been shown to affect four characteristic behaviors in rats, collectively known as Tetrad. For a description of these behaviors, see: PB Smith et al., The pharmacological activity of anandamide, a putative endogenous cannabinoid, in mice. in J. Pharmacol. Exp. Ther., 270 (1) (1994) 219-227, and J. Wiley et al., Discriminative stimulus effects of anandamide in rats in Eur. J. Pharmacol., 276 (1-2) (1995) 49-54. The reversal of these activities in the Locomotor Activity, Catalepsy, Hypothermia and Hot Plate assays, described below, provides an examination for the in vivo activity of CB-1 antagonists.

All data are presented as% reversal of isolated agonist using the following formula:

(5- (1,1-dimethyl-heptyl) -2- [5-hydroxy-2- (3-hydroxypropyl) -cyclohexyl] -phenol / agonist - vehicle / agonist) / (vehicle / vehicle - vehicle / agonist). Negative numbers indicate potentiation of antagonistic or non-antagonistic activity. The positive numbers indicate a reversal of activity for that particular test.

Locomotive Activity

Male ICR rats (n = 6; 17-19 g, Charles River Laboratories, Inc., Wilmington, MA) were pretreated with test compound (sc, po, ip or icv). Fifteen minutes later, the rats were stimulated with (5- (1,1dimethylheptyl) -2- [5-hydroxy-2- (3-hydroxypropyl) -cyclohexyl] -phenol (sc). Twenty-five minutes after agonist injection, the rats were placed in clear acrylic cages (431.8 cm x 20.9 cm x 20.3 cm), containing clean wood chips. Subjects were allowed to explore the surroundings for a total of 5 minutes and the activity was recorded by infrared motion detectors (available from Coulboum Instruments ™, Allentown, PA), which were placed on top of the cages. The data were collected by computer and expressed as units of movement. Catalepsy

Male ICR rats (n = 6; 17-19 g on arrival) were pre-treated with test compound (sc, po, ip or icv). Fifteen minutes later, the rats were stimulated with (5- (1,1-dimethyl-heptyl) -2- [5-hydroxy-2- (3-hydroxypropyl) -cyclohexyl] -phenol (sc). Ninety minutes after the injection of agonist, the rats were placed on a 6.5 cm steel ring attached to a ring support at a height of about 30 centimeters (12 inches). The ring was mounted in a horizontal orientation and the rat was suspended in the opening of the ring with the front and rear legs grasping the perimeter. The duration for which the rat remained completely immobile (except for breathing movements) was recorded over a period of 3 minutes.

The data were presented in the form of a classification of the percentage immobility. The classification was calculated by dividing the number of seconds in which the rat remains immobile by the total time of the observation period and multiplying the result by 100. A percentage reversal of the agonist was then calculated.

Hypothermia

Male ICR rats (n = 5; 17-19 g on arrival) were pre-treated with test compounds (sc, po, ip or icv). Fifteen minutes later, the rats were stimulated with the cannabinoid agonist (5- (1,1-dimethylheptyl) -2- [5-hydroxy-2- (3-hydroxypropyl) -cyclohexyl] -phenol (sc). five minutes after the 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

Male ICR rats (n = 7; 17-19 g on arrival) were pre-treated with test compounds (sc, po, ip or icv). Fifteen minutes later, the rats were stimulated with a cannabinoid agonist (5- (1,1-dimethylheptyl) -2- [5-hydroxy-2- (3-hydroxypropyl) -cyclohexyl] -phenol (sc). five minutes later, each rat was tested for reversal of analgesia using a standard hot plate meter (Columbus Instruments) The hot plate was 10x10x0.75 with a clear acrylic wall surrounding it. Latency for kicking, licking or shaking the hind leg, or jumping off the platform, was recorded to the nearest tenth of a second. The timekeeper was an activated experimenter and each test had a cut-off time of 40 seconds. The data were presented as a percentage reversal of agonist-induced analgesia.

Food Entrance

The following test was used to assess the effectiveness of the test compounds for inhibiting food intake in SpragueDawley rats after an overnight fast.

Male Sprague-Dawley rats were obtained from Charles River Laboratories, Inc. (Wilmington, MA). The rats were housed individually and fed with powdered food. They were maintained on a 12-hour light / dark cycle and received food and water ad libitum. The animals were acclimated to the nursery for a period of one week before testing was conducted. Testing was completed during the light portion of the cycle.

To conduct the food intake efficacy test, the rats were transferred to individual test cages, without food, the afternoon before testing, and the rats were fasted overnight. After fasting overnight, the rats were dosed the next morning with vehicle or test compounds. A known antagonist was dosed (3 mg / kg) as a positive control and a control group received an isolated vehicle (without compound). The test compounds were dosed at intervals between 0.1 and 100 mg / kg, depending on 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 have been used to accommodate various compounds, when required. The food was provided to the rats 30 minutes after dosing and the Oxymax automated food intake system (Columbus Instruments, Columbus, Ohio) was started. The entry of individual rat food was continuously recorded at 10 minute intervals over a period of two hours. When required, food intake was recorded manually, using an electronic scale; the food was weighed every 30 minutes after the food was provided, up to four hours after the food was provided. The effectiveness of the compound was determined by comparing the food intake pattern of the compound-treated rats with the vehicle and the standard positive control.

Alcohol Entry

The following protocol assesses the effects of alcohol intake in female rats who prefer alcohol (P) (raised at Indiana University) with an extensive drinking history. The following references provide detailed descriptions of the P mice: T.-K. Li et al., Indiana selection studies on alcohol related behaviors in Development of Animal Models as Pharmacogenetic Tools, CE McCIearn, RA Deitrich and VG Erwin (eds.), Research Monograph, 6 (1981) 171-192 NIAAA, ADAMHA, Rockville, MD; L. Lumeng et al., New strains of rats with alcohol preference and nonpreference in Alcohol And Aldehyde Metabolizing Systems, 3 (1977) 537-544, Academic Press, New York; and L. Lumeng et al., Different sensitivities to ethanol in alcohol-preferring and -nonpreferring rats in Pharmacol. Biochem. Behav., 16 (1982) 125-130.

Female rats were given 2 hours of access to alcohol (10% v / v and water, 2 bottles of their choice), daily, at the beginning of the cycle in the dark. The rats were kept on a reverse cycle to facilitate the experimenter's interactions. The animals were initially assigned to four equalized groups for alcohol intake: Group 1 - vehicle (n = 8); Group 2 - positive control (for example, 5.6 mg / kg AM251; n = 8); Group 3 - low dose of test compound (n = 8); and Group 4 - high dose of test compound (n = 8). The test compounds were generally mixed in a β93 cyclodextrin 30% (w / v) vehicle in distilled water, in a volume of 1-2 ml / kg. Vehicle injections were given to all groups during the first two days of the experiment. This was followed by 2 days of drug injections (for the appropriate groups) and a final day of vehicle injections. On the days of the drug injection, the drugs were given sc 30 minutes before a 2 hour alcohol access period. The intake of alcohol for all animals was measured during the test period and a comparison was made between animals treated with drugs and vehicles, to determine the effects of the compounds on alcohol drinking behavior.

Further studies on the drink have been done using female C57BI / 6 rats (Charles River). Several studies have shown that this strain of mice will readily consume alcohol with little to no handling required (Middaugh et al., Ethanol Consumption by C57BL / 6 Mice: Influence of Gender and Procedural Variables in Alcohol, 17 (3) (1999) 175 -183; Le et al., Alcohol Consumption by C57BL / 6, BALA / c and DBA / 2 Mice in a Limited Access Paradigm in Pharmacology Biochemistry and Behavior, 47 (1994) 375-378).

For our purposes, the rats (17-19 g), on arrival, were housed individually and with unlimited access to powdered rat food, 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 of access daily. This was done in a way that the access period was the last 2 hours of the dark part of the light cycle.

Once the drinking behavior stabilized, testing began. The rats were considered stable when the average consumption of alcohol for 3 days was ± 20% of the average for all 3 days. Day 1 of the test consisted of all rats receiving vehicle injection (sc or ip). Thirty to 120 minutes post-injection, access to alcohol and water was given. Alcohol consumption for that day was calculated (g / kg) and the groups were organized (n = 7-10) so that all groups had questionable alcohol intake. On days 2 and 3, the rats were injected with a vehicle or drug and the same protocol as the previous day was followed.

Day 4 was an internal wash and no injections were given. The data were analyzed using repeated ANOVA measures. The change in water and alcohol consumption was compared with the vehicle for each day of the test. The positive results should be interpreted as a compound that was able to significantly reduce alcohol consumption, to a stage that has no effect on water.

Oxygen Consumption

Methods:

Overall body oxygen consumption is measured using an indirect calorimeter (Oxymax from Columbus Instruments, Columbus, OH) in male Sprague Dawley rats (if another strain of rats or female rats is used, this will be specified). The rats (300-380 g of body weight) are placed in chambers with calorimeters and the chambers are placed on activity monitors. These studies are done during the light cycle. Before measuring oxygen consumption, rats are fed ad libitum with standard food. When measuring oxygen consumption, food is not available. Baseline 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 catheterization (or another route of administration as specified, that is, sc, ip, iv). The drugs are prepared in methylcellulose, water or another specified vehicle (examples include PEG400, beta-cyclodextran 30% and propylene glycol). Oxygen consumption and ambulatory activity are measured every 10 minutes, for an additional 1-6 hours post-dosing.

The Oxymax calorimeter program calculates oxygen consumption (mL / kg / h) based on the speed of air flow through the chambers and the difference in oxygen content at the inlet and outlet ports. Activity monitors have 15 beams of infrared light spaced 2.54 centimeters (one inch) on each axis, ambulatory activity is recorded when two consecutive beams are broken and the results are recorded as counts (or pulses).

Oxygen consumption at rest, during pre- and post-dosing, is calculated by averaging the consumption values ​​of O₂ 10 min, excluding periods of high ambulatory activity (ambulatory activity count&gt; 100) and excluding the first 5 values ​​of the pre-dose period and the first value of the post-dose period. The change in oxygen consumption is reported as a percentage and is calculated by dividing the consumption of oxygen at rest after dosing by the consumption of pre-dose oxygen x 100. Experiments will typically be done with n = 4-6 rats and the results reported are the mean ± SEM.

Interpretation:

An increase in oxygen consumption of&gt; 10% is considered a positive result. Historically, vehicle-treated rats have had no change in oxygen consumption from baseline pre-dose.

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