HK1213559B

HK1213559B - Pyrrolo[2,3-d]pyrimidine derivatives as cb2 receptor agonists

Info

Publication number: HK1213559B
Application number: HK16101493.2A
Authority: HK
Inventors: 尤伟．格雷瑟; 金原笃; 马蒂亚斯．内特科文; 斯蒂芬．勒韦尔; 马克．罗杰斯-埃文斯; 坦贾．舒尔兹-加施
Original assignee: 豪夫迈‧罗氏有限公司
Priority date: 2013-05-02
Filing date: 2014-04-29
Publication date: 2019-01-25

Description

Pyrrolo [2, 3-d ] pyrimidine derivatives as CB2 receptor agonists

The present invention relates to organic compounds useful for therapy and/or prophylaxis in mammals, and in particular to compounds which are preferential agonists of cannabinoid receptor 2.

The invention relates in particular to compounds of the formula (I),

wherein

A is CH₂Or is absent;

R¹and R²Together with the nitrogen atom to which they are attached form a substituted pyrrolidinyl or 2-oxa-6-azaspiro [3.3](vii) heptyl, wherein substituted pyrrolidinyl is pyrrolidinyl substituted with one or two substituents independently selected from the group consisting of halogen, hydroxy, and alkylcarbonyloxy; and is

R³Is halophenyl, alkylsulfonylphenyl, haloalkylphenyl, halopyridyl, alkylOxadiazolyl, alkyltriazolyl, alkyltetrazolyl, tetrahydrofuranyl (oxolanyl), cycloalkyltetrazolyl or haloalkyl-1H-pyrazolyl;

or a pharmaceutically acceptable salt or ester thereof.

The compounds of formula (I) are particularly useful for the treatment or prophylaxis of, for example, pain (pain), atherosclerosis (atherosclerosis), age-related macular degeneration, diabetic retinopathy (diabetic retinopathy), glaucoma (glaucoma), retinal vein occlusion (chronic retinal occlusion), retinopathy of prematurity (retinopathy of prematurity), ocular ischemic syndrome (ischemic syndrome), geographic atrophy (geographic atrophy), diabetes mellitus (diabetes mellitus), inflammation (inflammation), inflammatory bowel disease (inflammatory bowel disease), ischemia-reperfusion injury (ischemic-reperfusion injury), acute failure (acute liver failure), liver fibrosis (fibrosis), acute kidney failure (acute kidney transplant), chronic renal fibrosis (fibrosis), diabetic nephropathy (diabatic nephopathopathy), glomerulonephropathy (glomeronephopathopathy), cardiomyopathy (cardiomyopopathopathy), heart failure (heart failure), myocardial ischemia (myo-ischemic), myocardial infarction (myo-ischemic attack), systemic sclerosis (systemic sclerosis), thermal injury (thermal injury), burn (burning), hypertrophic scars (hypertrophic scars), keloids (keloids), gingivitis pyrexia, liver cirrhosis (liver cirrhosis) or tumors (tumors), regulation of bone mass (bone), amyotrophic lateral sclerosis (amyotropic sclerosis), multiple sclerosis (multiple sclerosis), Alzheimer's disease, Parkinson's disease, stroke (ischemic stroke), uveitis, multiple sclerosis (Parkinson's disease), Parkinson's disease, stroke (ischemic stroke).

The compounds of formula (I) are particularly useful for the treatment or prevention of diabetic retinopathy, retinal vein occlusion or uveitis.

Cannabinoid receptors are a class of cell membrane receptors belonging to the G protein-coupled receptor superfamily. There are currently two known subtypes, known as cannabinoid receptor 1(CB1) and cannabinoid receptor 2(CB 2). The CB1 receptor is expressed predominantly in the central nervous (i.e. amygdala cerebellum, hippocampus) system and in lesser amounts in the periphery. CB2 encoded by the CNR2 gene is expressed predominantly on cells of the immune system, such as macrophages and T-cells (Ashton, J.C. et al, Curr Neuropharmacol 2007, 5(2), 73-80; Miller, A.M. et al, Br J Pharmacol 2008, 153(2), 299-308; Centonze, D., et al, Curr PharmDes 2008, 14(23), 2370-42), and peripherally in the gastrointestinal system (Wright, K.L. et al, Br J Pharmacol 2008, 153(2), 263-70). The CB2 receptor is also widely distributed in the brain, where it is found primarily on microglia rather than neurons (Cabral, g.a. et al Br J Pharmacol 2008, 153 (2): 240-51).

Interest in agonists of the CB2 receptor has steadily increased over the past decade (there are currently 30-40 patent applications/year) due to the fact that several of the early compounds have been shown to have beneficial effects in preclinical models of many human diseases, including chronic pain (Beltramo, m.mini Rev Med Chem 2009, 9(1), 11-25), atherosclerosis (Mach, f. et al, J neuroendicrinol 2008, 20Suppl 1, 53-7), regulation of bone mass (Bab, i. et al, Br J Pharmacol 2008, 153(2), 182-8), neuroinflammation (Cabral, g.a. et al, J leocalcic biol, 78(6), 1192-7), ischemia/reperfusion injury (pocher, p. et al, Br J Pharmacol, 153(2), 252-62), systemic fibrosis (akhmistina, rhesus et al, 2009, 60, (4), 1129-36; Garcia-Gonzalez, e, et al, rheumatology (oxford)2009, 48(9), 1050-6), liver fibrosis (Julien, b, et al, Gastroenterology 2005, 128(3), 742-55; Munoz-Luque, J, et al, J Pharmacol ExpTher 2008, 324(2), 475-83).

Ischemia/reperfusion (I/R) injury is the leading cause of tissue damage that occurs in conditions such as stroke, myocardial infarction, cardiopulmonary bypass and other vascular procedures, and organ transplantation, and is the major mechanism of end organ damage that complicates the circulatory shock process of various etiologies. All of these disorders are characterized by an interruption of normal blood supply, resulting in inadequate tissue oxygenation. Reoxygenation, e.g. reperfusion, is the final treatment to restore normal tissue oxygenation. But the lack of oxygen and nutrients from the blood produces a condition in which the restoration of circulation leads to further tissue damage. Reperfusion injury is caused in part by the inflammatory response of the damaged tissue. Leukocytes transported to the area by the newly regurgitated blood release large amounts of inflammatory factors such as interleukins and free radicals in response to tissue damage. The restored blood flow reintroduces intracellular oxygen, which damages cellular proteins, DNA, and plasma membranes.

Remote Ischemic Preconditioning (RIPC) represents a strategy to exploit the body's endogenous protective capacity against damage caused by ischemia and reperfusion. It describes the interesting phenomenon where transient non-lethal ischemia and reperfusion of one organ or tissue confers resistance to subsequent events of "lethal" ischemia reperfusion injury in distant organs or tissues. Despite several hypotheses, the actual mechanism by which transient ischemia and reperfusion of an organ or tissue confers protection is currently unknown.

The humoral hypothesis suggests various intracellular pathways for endogenous substances produced in distant organs or tissues (such as adenosine, bradykinin, opioids, CGRP, endocannabinoids, angiotensin I or some other not yet identified humoral factors) to enter the bloodstream and activate their respective receptors in the target tissues and thereby restore cardioprotection involved in ischemic preconditioning.

Recent data indicate that endocannabinoids and their receptors, in particular CB2, may be involved in pretreatment and contribute to the prevention of reperfusion injury through down-regulation of the inflammatory response (pocher, p. et al, Br J Pharmacol 2008, 153(2), 252-62). In particular, recent studies using CB2 tool agonists have shown the efficacy of this concept for reducing I/R injury in the heart (Defer, n. et al, Faseb J2009, 23(7), 2120-30), brain (Zhang, m. et al, J Cereb Blood FlowMetab 2007, 27(7), 1387-96), liver (Batkai, s. et al, Faseb J2007, 21(8), 1788-.

Furthermore, over the past years, increasing literature has shown that CB2 may also be of interest in subchronic and chronic situations. Specific up-regulation of CB1 and CB2 has been shown to be associated with CB 2-related expression in myofibroblasts, i.e. cells that are responsible for the progress of fibrosis, in animal models of chronic diseases associated with fibrosis (Garcia-Gonzalez, e. et al, rheumatology (oxford)2009, 48(9), 1050-6; Yang, y. et al, lever Int2009, 29(5), 678-85).

Activation of the CB2 receptor by selective CB2 agonists has in fact been shown to produce an anti-fibrotic effect in diffuse systemic sclerosis (Garcia-Gonzalez, e. et al, rhematology (oxford)2009, 48(9), 1050-6) and the CB2 receptor has been shown to be a key target in experimental dermal fibrosis (Akhmetshina, a. et al, Arthritis Rheum 2009, 60(4), 1129-36) and in liver pathophysiology, including fibrogenesis associated with chronic liver disease (Lotersztajn, s. et al, gasstrenter Clin Biol, 2007 (3), 255-8; Mallat, a. et al, Expert Opin Targets2007, 11(3), 403-9; Lotersztajn, s. et al, Br J2008, Pharmacol (2), 286-9).

The compounds of the present invention bind to and modulate the CB2 receptor and have reduced CB1 receptor activity.

In the present specification, the term "alkyl", alone or in combination, denotes a straight-chain or branched alkyl group having 1 to 8 carbon atoms, in particular a straight-chain or branched alkyl group having 1 to 6 carbon atoms, more in particular a straight-chain or branched alkyl group having 1 to 4 carbon atoms. Straight and branched C₁-C₈Examples of alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, the isomeric pentyl groups, the isomeric hexyl groups, the isomeric heptyl groups and the isomeric octyl groups, in particular methyl, ethyl, propyl, butyl and pentyl groups. A particular example of an alkyl group is methyl.

The term "cycloalkyl", alone or in combination, denotes a cycloalkyl ring having 3 to 8 carbon atoms, and in particular a cycloalkyl ring having 3 to 6 carbon atoms. Examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl, cycloheptyl and cyclooctyl. A particular example of a cycloalkyl group is cyclopropyl.

The term "halogen" or "halo", alone or in combination, denotes fluorine, chlorine, bromine or iodine and especially fluorine, chlorine or bromine, more especially fluorine and chlorine. The term "halo", in combination with another group, means the substitution of the group by at least one halogen, in particular by one to five halogens, in particular one to four halogens, i.e. one, two, three or four halogens. Particular "halogen" are fluorine and chlorine. At R¹And R²In the definition of (1), fluorine is a particular halogen.

The term "haloalkyl", alone or in combination, denotes an alkyl substituted by at least one halogen, in particular by one to five halogens, in particular one to three halogens. A particular "haloalkyl" is trifluoromethyl.

The terms "hydroxy" and "hydroxyl", alone or in combination, denote an-OH group.

The term "carbonyl", alone or in combination, denotes a-c (o) -group.

The term "oxo", alone or in combination, denotes an-O-group.

The term "amino", alone or in combination, denotes a primary amino group (-NH)₂) A secondary amino group (-NH-), or a tertiary amino group (-N-). A particular amino group is-NH-.

The term "sulfonyl", alone or in combination, denotes-S (O)₂-a group.

The term "pharmaceutically acceptable salts" refers to those salts that retain the biological effectiveness and properties of the free base or free acid, and which are not biologically or otherwise undesirable. The salts are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, in particular hydrochloric acid, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine. In addition, these salts can be prepared by the addition of an inorganic or organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium salts. Salts derived from organic bases include, but are not limited to, salts of: primary, secondary and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyamine resins. The compounds of formula (I) may also be present in zwitterionic form. Particularly preferred pharmaceutically acceptable salts of the compounds of formula (I) are salts of hydrochloric, hydrobromic, sulfuric, phosphoric and methanesulfonic acids.

By "pharmaceutically acceptable ester" is meant that the compound of formula (I) may be derivatised at functional groups to provide derivatives which are capable of conversion back to the parent compound in vivo. Examples of such compounds include physiologically acceptable and metabolically labile ester derivatives such as methoxymethyl ester, methylthiomethyl ester and pivaloyloxymethyl ester. Furthermore, any physiologically acceptable equivalent of a compound of formula (I) that is similar to the metabolically labile ester and is capable of producing the parent compound of formula (I) in vivo is within the scope of the invention.

If one of the starting materials or the compound of formula (I) contains one or more functional Groups which are unstable or reactive under the reaction conditions of one or more reaction steps, suitable protecting Groups can be introduced before the critical step using methods known in the art (as described, for example, in T.W.Greene and P.G.M.Wuts in "Protective Groups in organic chemistry", 3 rd edition, 1999, Wiley, New York). Such protecting groups can be removed at a later stage of the synthesis using standard methods described in the literature. Examples of protecting groups are t-butyloxycarbonyl (Boc), 9-fluorenylmethyl carbamate (Fmoc), 2-trimethylsilylethyl carbamate (Teoc), benzyloxycarbonyl (Cbz) and p-methoxybenzyloxycarbonyl (Moz).

The compounds of formula (I) may contain several asymmetric centers and may be present in the form of optically pure enantiomers, mixtures of enantiomers such as, for example, racemates, mixtures of diastereoisomers, diastereoisomeric racemates or mixtures of diastereoisomeric racemates.

The term "asymmetric carbon atom" means a carbon atom having four different substituents. According to Cahn-Ingold-Prelog Convention, the asymmetric carbon atoms may be in either the "R" or "S" configuration.

The invention relates in particular to:

a compound of formula (I) wherein A is CH₂；

A compound of formula (I) wherein R¹And R²Together with the nitrogen atom to which they are attached form a substituted pyrrolidinyl group, wherein substituted pyrrolidinyl is pyrrolidinyl substituted with one or two substituents independently selected from halogen and hydroxy;

a compound of formula (I) wherein R¹And R²Together with the nitrogen atom to which they are attached form a substituted pyrrolidinyl group, wherein substituted pyrrolidinyl is pyrrolidinyl substituted with one or two substituents independently selected from fluoro and hydroxy;

a compound of formula (I) wherein R¹And R²Together with the nitrogen atom to which they are attached form difluoropyrrolidinyl or hydroxypyrrolidinyl;

a compound of formula (I) wherein R¹And R²Together with the nitrogen atom to which they are attached form difluoropyrrolidinyl, 2-oxa-6-azaspiro [3.3]Heptyl, hydroxypyrrolidinyl, or methylcarbonyloxypyrrolidinyl;

a compound of formula (I) wherein R³Is halophenyl, haloalkylphenyl, alkylsulfonylphenyl, halopyridyl or alkylA diazolyl group;

a compound of formula (I) wherein R³Is dichlorophenyl, chlorophenyl, trifluoromethylphenyl, methylsulfonylphenyl, chloropyridyl or methylA diazolyl group; and

a compound of formula (I) wherein R³Is chlorophenyl, dichlorophenyl, chlorophenyl, methylsulfonylphenyl, trifluoromethylphenyl, chloropyridyl, methylOxadiazolyl, dimethyltriazolyl, methyltetrazolyl, cyclopropyltetrazolyl, tetrahydrofuryl or trifluoromethyl-1H-pyrazolyl.

The invention also relates to a compound of formula (I) selected from:

2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine;

2-tert-butyl-7- [ (2-chloro-4-fluorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine;

2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) -7- [ (2-methanesulfonylphenyl) methyl ] pyrrolo [2, 3-d ] pyrimidine;

2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) -7- [ [2- (trifluoromethyl) phenyl ] methyl ] pyrrolo [2, 3-d ] pyrimidine;

2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine;

2-tert-butyl-7- [ (2-chloropyridin-3-yl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine;

2-tert-butyl-7- [ (3-chloropyridin-2-yl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine;

5- [ [ 2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d]Pyrimidin-7-yl]Methyl radical]-3-methyl-1, 2, 4-Oxadiazole;

3- [ [ 2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d]Pyrimidin-7-yl]Methyl radical]-4-methyl-1, 2, 5-Oxadiazole;

2- [ [ 2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d]Pyrimidin-7-yl]Methyl radical]-5-methyl-1, 3, 4-Oxadiazole;

2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) -7- [ (4, 5-dimethyl-1, 2, 4-triazol-3-yl) methyl ] pyrrolo [2, 3-d ] pyrimidine;

2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) -7- [ (1-methyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidine;

2-tert-butyl-7- [ (1-cyclopropyltetrazol-5-yl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine;

6- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane;

6- [ 2-tert-butyl-7- [ (3-methyl-1, 2, 4-Oxadiazol-5-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]-2-oxa-6-azaspiro [3.3]Heptane;

6- [ 2-tert-butyl-7- [ (3S) -tetrahydrofuran-3-yl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane;

6- [ 2-tert-butyl-7- [ (3R) -tetrahydrofuran-3-yl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane;

6- [ 2-tert-butyl-7- [ (2-chloro-4-fluorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane;

6- [ 2-tert-butyl-7- [ (2-methanesulfonylphenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane;

6- [ 2-tert-butyl-7- [ (2-chloropyridin-3-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane;

6- [ 2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane;

6- [ 2-tert-butyl-7- [ (4-methyl-1, 2, 5-)Oxadiazol-3-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]-2-oxa-6-azaspiro [3.3]Heptane;

6- [ 2-tert-butyl-7- [ (5-methyl-1, 3, 4-Diazoles-2-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]-2-oxa-6-azaspiro [3.3]Heptane;

1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (2-chloro-4-fluorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (2-methanesulfonylphenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ [2- (trifluoromethyl) phenyl ] methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (2-chloropyridin-3-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (3-chloropyridin-2-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (3-methyl-1, 2, 4-Oxadiazol-5-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (4-methyl-1, 2, 5-)Oxadiazol-3-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (5-methyl-1, 3, 4-Oxadiazol-2-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (1-methyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (1-methyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-yl ] acetate;

1- [ 2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-yl ] acetate;

acetic acid [1- [ 2-tert-butyl-7- [ (4-methyl-1, 2, 5-)Oxadiazol-3-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-yl radical]An ester;

1- [ 2-tert-butyl-7- [ (1-cyclopropyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ (3R) -tetrahydrofuran-3-yl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

1- [ 2-tert-butyl-7- [ [3- (trifluoromethyl) -1H-pyrazol-4-yl ] methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) -7- [ [3- (trifluoromethyl) -1H-pyrazol-4-yl ] methyl ] pyrrolo [2, 3-d ] pyrimidine;

(3S) -1- [ 2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

(3S) -1- [ 2-tert-butyl-7- [ (2-chloro-4-fluorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

(3S) -1- [ 2-tert-butyl-7- [ [2- (trifluoromethyl) phenyl ] methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

(3S) -1- [ 2-tert-butyl-7- [ (2-methanesulfonylphenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

(3S) -1- [ 2-tert-butyl-7- [ (2-chloropyridin-3-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

(3S) -1- [ 2-tert-butyl-7- [ (3-chloropyridin-2-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol;

(3S) -1- [ 2-tert-butyl-7- [ (4-methyl-1, 2, 5-)Oxadiazol-3-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-ol;

6- [ 2-tert-butyl-7- [ (3-chloropyridin-2-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane; and

6- [ 2-tert-butyl-7- [ (1-methyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane.

The invention also relates to a compound of formula (I) selected from:

(3S) -1- [ 2-tert-butyl-7- [ (3-chloropyridin-2-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol; and

(3S) -1- [ 2-tert-butyl-7- [ (4-methyl-1, 2, 5-)Oxadiazol-3-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-ol.

The synthesis of the compounds of formula (I) can be achieved, for example, according to the following scheme.

Unless otherwise stated, A and R¹To R³Have the meaning as defined above.

The preparation of the compounds of formula I according to the invention can be carried out in a continuous or convergent (convergent) synthetic route. In the following schemes, the synthesis of the compounds of the present invention is shown. The skills required to carry out the reactions and purification of the resulting products are well known to those skilled in the art. Unless indicated to the contrary, the substituents and symbols used in the following process descriptions have the meanings given herein before. In more detail, the compounds of formula I can be manufactured by the methods given below, by the methods given in the examples or by analogous methods. Suitable reaction conditions for the individual reaction steps are well known to the person skilled in the art. Also, for conditions described in the literature that affect the reaction, see, for example: comprehensive organic transformation: a guide to the preparation of Functional groups (Comprehensive Organic Transformations: AGuide to Functional Group precursors), 2 nd edition, Richard C.Larock, John Wiley & Sons, New York, NY.1999). We have found that it is convenient to carry out the reaction in the presence or absence of a solvent. There is no particular restriction on the nature of the solvent used, provided that it has no adverse effect on the reaction or the reagents involved and that it can dissolve the reagents to at least some extent. The reaction can be carried out over a wide temperature range and the precise reaction temperature is not critical to the invention. It is convenient to carry out the reaction at a temperature in the range of-78 ℃ to reflux. The time required for the reaction may also vary widely, depending on a number of factors, notably on the reaction temperature and the nature of the reagents. However, a period of from 0.5h to several days will generally be sufficient to produce the intermediates and compounds. The reaction sequence is not limited to the reaction sequence shown in the scheme, but the order of the reaction steps may be freely changed depending on the raw materials and their respective reactivities. Raw materials are commercially available or can be prepared by methods similar to those given below, by methods described in references cited in the specification or in the examples, or by methods well known in the art.

Scheme 1

a) 2-tert-butyl-4-chloro-7H-pyrrolo [2, 3-d ] pyrimidine II is commercially available and can be conveniently reacted with an amine (commercially available, or well known in the art) in the presence or absence of a base to provide intermediate III.

b) Intermediate III may be conveniently reacted with an electrophile (commercially available, or well known in the art) in the presence or absence of a base to give the title compound (I).

Any protecting group used in this sequence may be cleaved after either of steps a) or b).

The present invention therefore also relates to a process for the preparation of a compound of formula (I), which process comprises reacting a compound of formula (A) at X-A-R³Reaction in the presence of:

wherein X is a leaving group and wherein A and R¹To R³Is as defined above.

In the process of the invention, suitable leaving groups are, for example, chlorine or bromine.

The process of the invention may be carried out in the presence of a base. Examples of suitable bases are NaH or KOtBu.

The process of the invention can be carried out, for example, in NMP (N-methyl-2-pyrrolidone), DMF (dimethylformamide) or THF (tetrahydrofuran).

The invention also relates to compounds of formula (I) prepared according to the above process.

The invention also relates in particular to:

the compounds of formula (I) are useful in the treatment or prophylaxis of pain, atherosclerosis (atherosclerosis), age-related macular degeneration (age-related macular degeneration), diabetic retinopathy (diabetic retinopathy), glaucoma (glaucoma), retinal vein occlusion (diabetic retinopathy), retinopathy of prematurity (retinopathy of prematurity), ocular ischemic syndrome (ocular ischemic syndrome), geographic atrophy (geographic phtopathy), diabetes mellitus (diabetes mellitus), inflammation (inflammation), inflammatory bowel disease (inflammatory bowel disease), ischemia-reperfusion injury (ischemic-reperfusion injury), acute liver failure (acute liver failure), liver fibrosis (fibrosis), lung fibrosis (fibrosis), fibrosis (fibrosis), acute kidney disease (acute kidney disease), chronic renal fibrosis (fibrosis), chronic diabetic nephropathy (fibrosis), glomerulonephropathy (glomeronephpathgy), cardiomyopathy (cardiomyopathiy), heart failure (heart failure), myocardial ischemia (myotopographic ischemia), myocardial infarction (myotopographic interaction), systemic sclerosis (systemic sclerosis), thermal injury (thermal injury), burn (burning), hypertrophic scars (hypertrophic scars), keloids (keloids), gingivitis pyrexia (gingivitis), liver cirrhosis (liver cirrhosis) or tumors, regulation of bone mass, amyotrophic lateral sclerosis (amylotrophic sclerosis), multiple sclerosis (multiple sclerosis), Alzheimer's disease, Parkinson's disease, stroke (ischemic stroke), stroke (stroke);

the use of a compound according to formula (I) in the manufacture of a medicament for the treatment or prophylaxis of pain, atherosclerosis, age-related macular degeneration, diabetic retinopathy, glaucoma, retinal vein occlusion, retinopathy of prematurity, ocular ischemic syndrome, geographic atrophy, diabetes mellitus, inflammation, inflammatory bowel disease, ischemia-reperfusion injury, acute liver failure, liver fibrosis, lung fibrosis, kidney fibrosis, systemic fibrosis, acute allograft rejection, chronic allograft nephropathy, diabetic nephropathy, glomerulonephropathy, cardiomyopathy, heart failure, myocardial ischemia, myocardial infarction, systemic sclerosis, thermal injury, burning, hypertrophic scars, keloids, gingivitis pyrexia, liver cirrhosis or tumors, regulation of bone mass, amyotrophic lateral sclerosis, multiple sclerosis, alzheimer's disease, parkinson's disease, stroke, transient ischemic attack or uveitis;

a compound of formula (I) for use in the treatment or prophylaxis of pain, atherosclerosis, age-related macular degeneration, diabetic retinopathy, glaucoma, retinal vein occlusion, retinopathy of prematurity, ocular ischemic syndrome, geographic atrophy, diabetes mellitus, inflammation, inflammatory bowel disease, ischemia-reperfusion injury, acute liver failure, liver fibrosis, lung fibrosis, kidney fibrosis, systemic fibrosis, acute allograft rejection, chronic allograft nephropathy, diabetic nephropathy, glomerulonephropathy, cardiomyopathy, heart failure, myocardial ischemia, myocardial infarction, systemic sclerosis, thermal injury, burning, hypertrophic scars, keloids, gingivitis pyrexia, liver cirrhosis or tumors, regulation of bone mass, amyotrophic lateral sclerosis, multiple sclerosis, alzheimer's disease, parkinson's disease, stroke, transient ischemic attack or uveitis; and

a method for the treatment or prophylaxis of pain, atherosclerosis, age-related macular degeneration, diabetic retinopathy, glaucoma, retinal vein occlusion, retinopathy of prematurity, ocular ischemic syndrome, geographic atrophy, diabetes mellitus, inflammation, inflammatory bowel disease, ischemia-reperfusion injury, acute liver failure, liver fibrosis, lung fibrosis, kidney fibrosis, systemic fibrosis, acute allograft rejection, chronic allograft nephropathy, diabetic nephropathy, glomerulonephropathy, cardiomyopathy, heart failure, myocardial ischemia, myocardial infarction, systemic sclerosis, thermal injury, burning, hypertrophic scars, keloids, gingivitis pyrexia, liver cirrhosis or tumors, regulation of bone mass, amyotrophic lateral sclerosis, multiple sclerosis, Alzheimer's disease, Parkinson's disease, stroke, transient ischemic attack or uveitis, the method comprises administering to a patient in need thereof an effective amount of a compound of formula (I).

The invention relates in particular to compounds of formula (I) for use in the treatment or prevention of ischemia, reperfusion injury, liver fibrosis or kidney fibrosis, in particular ischemia or reperfusion injury.

The invention relates in particular to compounds of formula (I) for use in the treatment or prevention of myocardial infarction.

The invention also relates in particular to compounds of formula (I) for use in the treatment or prevention of age-related macular degeneration, diabetic retinopathy, glaucoma, retinal vein occlusion, retinopathy of prematurity, ocular ischemic syndrome, geographic atrophy or uveitis.

The invention also relates in particular to compounds of formula (I) for use in the treatment or prevention of amyotrophic lateral sclerosis or multiple sclerosis.

The invention also relates to compounds of formula (I) prepared according to the process of the invention.

Another embodiment of the present invention provides pharmaceutical compositions or medicaments comprising a compound of the present invention and a therapeutically inert carrier, diluent or excipient, as well as methods of using the compounds of the present invention for preparing such compositions and medicaments. In one example, a compound of formula (I) may be formulated as follows: the galenical administration form is prepared by mixing at ambient temperature, at a suitable pH, and at the desired degree of purity, with a physiologically acceptable carrier, i.e. a carrier which is non-toxic to the recipient at the dosages and concentrations employed. The pH of the formulation depends primarily on the particular use and concentration of the compound, but is preferably anywhere in the range of about 3 to about 8. In one example, the compound of formula (I) is formulated in an acetate buffer at pH 5. In another embodiment, the compounds of formula (I) are sterile. The compounds may be stored, for example, as solid or amorphous compositions, as lyophilized formulations, or as aqueous solutions.

The compositions are formulated, dosed, and administered in a manner consistent with good medical practice. Factors to be considered herein include the particular condition being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the condition, the site of delivery of the agent, the method of administration, the timing of administration, and other factors known to the practitioner.

The compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if topical treatment is desired, intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

The compounds of the present invention may be administered in any convenient form of administration, for example, tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, and the like. Such compositions may contain conventional ingredients of pharmaceutical preparations such as diluents, carriers, pH adjusting agents, sweeteners, fillers, and other active agents.

Typical formulations are prepared by mixing a compound of the invention with a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail, for example, in Ansel, Howard c, et al, Ansel's pharmaceutical dosage Forms and Drug Delivery systems. Lippincott, Williams & Wilkins, 2004; gennaro, Alfonso r., et al Remington: the Science and Practice of pharmacy, Philadelphia: lippincott, Williams & Wilkins, 2000; and Rowe, Raymond. handbook of Pharmaceutical excipients. Chicago, Pharmaceutical Press, 2005. The formulations may also include one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifying agents (glidants), glidants, processing aids, colorants, sweeteners, flavoring agents, diluents, and other known additives to provide a superior presentation of the drug (i.e., a compound of the present invention or a pharmaceutical composition thereof) or to aid in the preparation of the pharmaceutical product (i.e., a pharmaceutical product).

The invention will now be illustrated by the following examples which are not to be construed as limiting in nature.

Examples

Example 1

2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine

a) 2-tert-butyl-1, 7-dihydro-pyrrolo [2, 3-d ] pyrimidin-4-one

To a solution of sodium ethoxide (210mL, 563 mmol; 21% in EtOH) in anhydrous EtOH (290mL) was added ethyl-2-cyano-4, 4-diethoxybutyrate ester (45g, 281mmol) and 2, 2-dimethyl-propionamidine (2, 2-dimethyl-propioamidine) (59g, 256mmol) and heated to reflux for 12 h. The mixture was concentrated, the residue diluted with water (100mL) and acidified with 2N aqueous HCl (pH 6-7). The precipitate was filtered, dried under vacuum and dissolved in EtOH (100 mL). Addition of H at 0 deg.C₂SO₄(10mL) and the mixture was heated to reflux for 2 h. After cooling to 25 ℃ with NH₃The aqueous solution made the solution alkaline (pH 8-9). The precipitate was filtered and dried under vacuum to give 2-tert-butyl-1, 7-dihydro-pyrrolo [2, 3-d as a white solid]Pyrimidin-4-one (18g, 47%) was used in the next step without further purification. MS (m/e): 192(M + H).

b) 2-tert-butyl-4-chloro-7H-pyrrolo [2, 3-d ] pyrimidine

2-tert-butyl-1, 7-dihydro-pyrrolo [2, 3-d)]Pyrimidin-4-one (2.5g, 13mmol) and POCl₃The mixture (10mL) was refluxed for 3 h. After cooling to 25 ℃, ice water (40mL) was added and the aqueous layer was extracted with EtOAc (2 × 75 mL). The combined organic layers were washed with water (4X 50mL) and brine (40mL) over anhydrous Na₂SO₄Dried, filtered and concentrated in vacuo. By column chromatographyThe residue was purified on flash silica gel (50% EtOAc/hexanes) to give the title compound as a light yellow solid after evaporation of the product containing fractions (1.2g, 44%). MS (m/e): 210.4(M + H).

c) 2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d ] pyrimidine

2-tert-butyl-4-chloro-7H-pyrrolo [2, 3-d]A solution of pyrimidine (1g, 4.7mmol), 3-difluoro-pyrrolidine hydrochloride (1.1g, 7.18mmol) and DIPEA (3mL, 14.1mmol) in EtOH (15mL) was stirred in a sealed tube at 100 ℃ for 16 h. The volatiles were removed in vacuo and the residue was dissolved in DCM (20 mL). The mixture was washed with water (2 × 10mL), brine (10mL) and over anhydrous Na₂SO₄Dried, filtered and evaporated under reduced pressure. The residue was purified by column chromatography on silica eluting with 20% EtOAc/hexanes to give the title compound as an off-white solid (1g, 74%). MS (m/e): 280.8(M + H).

d) 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine

2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d]A solution of pyrimidine (50mg, 0.17mmol), potassium tert-butoxide (31mg, 0.268mmol) and 2-chlorobenzyl bromide (55mg, 0.268mmol) in NMP (1mL) was heated under microwave irradiation at 150 ℃ for 30 min. The reaction mixture was cooled to 25 ℃, diluted with EtOAc (15mL), washed with water (2 × 10mL), over anhydrous Na₂SO₄Dried, filtered and evaporated under reduced pressure. The residue was purified by preparative HPLC [ Xterra-RP18, 10. mu.19X 250 mM/acetonitrile/10 mM, using ammonium acetate in water as solvent system]Purification to give the title compound as a colorless viscous solid after evaporation of the product containing fractions (30mg, 41%). MS (m/e): 405(M + H).

Example 2

2-tert-butyl-7- [ (2-chloro-4-fluorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine

In analogy to the procedure described for the synthesis of 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine (example 1) the title compound was prepared from 2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 1, step c). MS (m/e): 423(M + H).

Example 3

2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) -7- [ (2-methanesulfonylphenyl) methyl ] pyrrolo [2, 3-d ] pyrimidine

In analogy to the procedure described for the synthesis of 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine (example 1) the title compound was prepared from 2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 1, step c). MS (m/e): 449(M + H).

Example 4

2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) -7- [ [2- (trifluoromethyl) phenyl ] methyl ] pyrrolo [2, 3-d ] pyrimidine

In analogy to the procedure described for the synthesis of 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine (example 1) the title compound was prepared from 2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 1, step c). MS (m/e): 439(M + H).

Example 5

2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine

Example 6

2-tert-butyl-7- [ (2-chloropyridin-3-yl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine

In analogy to the procedure described for the synthesis of 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine (example 1) the title compound was prepared from 2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 1, step c). MS (m/e): 406(M + H).

Example 7

2-tert-butyl-7- [ (3-chloropyridin-2-yl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine

Example 8

5- [ [ 2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d]Pyrimidin-7-yl]Methyl radical]-3-methyl-1, 2, 4-Diazoles

In analogy to the procedure described for the synthesis of 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine (example 1) the title compound was prepared from 2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 1, step c). MS (m/e): 377(M + H).

Example 9

3- [ [ 2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d]Pyrimidin-7-yl]Methyl radical]-4-methyl-1, 2, 5-Diazoles

Example 10

2- [ [ 2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d]Pyrimidin-7-yl]Methyl radical]-5-methyl-1, 3, 4-Diazoles

Example 11

2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) -7- [ (4, 5-dimethyl-1, 2, 4-triazol-3-yl) methyl ] pyrrolo [2, 3-d ] pyrimidine

In analogy to the procedure described for the synthesis of 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine (example 1) the title compound was prepared from 2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 1, step c). MS (m/e): 390(M + H).

Example 12

2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) -7- [ (1-methyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidine

Example 13

2-tert-butyl-7- [ (1-cyclopropyltetrazol-5-yl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine

In analogy to the procedure described for the synthesis of 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine (example 1) the title compound was prepared from 2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 1, step c). MS (m/e): 403(M + H).

Example 14

6- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane

a) 2-tert-butyl-4- (2-oxa-6-aza-spiro [3.3] hept-6-yl) -7H-pyrrolo [2, 3-d ] pyrimidine

2-tert-butyl-4-chloro-7H-pyrrolo [2, 3-d]Pyrimidine (1g, 4.7mmol), 2-oxa-6-aza-spiro [ 3.3%]A solution of heptane oxalate (2g, 7.18mmol) and DIPEA (3mL, 14.1mmol) in EtOH (15mL) was stirred in a sealed tube at 100 ℃ for 16 h. The volatiles were removed in vacuo and the residue was dissolved in DCM (20 mL). The organic layer was washed with water (2 × 10mL) and brine (10mL) over anhydrous Na₂SO₄Dried, filtered, and evaporated. The residue was purified by column chromatography on silica eluting with 20% EtOAc/hexanes to give the title compound as a white solid (750mg, 57%). MS (m/e): 273(M + H).

b)6- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane

2-tert-butyl-4- (2-oxa-6-aza-spiro [3.3]]Hept-6-yl) -7H-pyrrolo [2, 3-d]A solution of pyrimidine (30mg, 0.11mmol), NaH (10mg, 0.132mmol) and bromomethyl-2-chloro-benzene (30mg, 0.143mmol) in DMF (5mL) was stirred at 25 ℃ for 12 h. Addition of NH₄Aqueous Cl solution and concentrated in vacuo. The residue was dissolved in water (10mL),and extracted with EtOAc (2 × 25 mL). The combined organic layers were washed with brine, over anhydrous Na₂SO₄Dried, filtered and evaporated. The residue was purified by preparative HPLC to give the title compound as an off-white viscous solid (14mg, 32%). MS (m/e): 397(M + H).

Example 15

6- [ 2-tert-butyl-7- [ (3-methyl-1, 2, 4-Oxadiazol-5-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]-2-oxa-6-azaspiro [3.3]Heptane (Heptane)

In analogy to the procedure described for the synthesis of 6- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane (example 14) the title compound was prepared from 2-tert-butyl-4- (2-oxa-6-aza-spiro [3.3] hept-6-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 14, step a). MS (m/e): 369(M + H).

Example 16

6- [ 2-tert-butyl-7- [ (3S) -tetrahydrofuran-3-yl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane

In analogy to the procedure described for the synthesis of 6- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane (example 14) the title compound was prepared from 2-tert-butyl-4- (2-oxa-6-aza-spiro [3.3] hept-6-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 14, step a). MS (m/e): 343(M + H).

Example 17

6- [ 2-tert-butyl-7- [ (3R) -tetrahydrofuran-3-yl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane

Example 18

6- [ 2-tert-butyl-7- [ (2-chloro-4-fluorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane

In analogy to the procedure described for the synthesis of 6- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane (example 14) the title compound was prepared from 2-tert-butyl-4- (2-oxa-6-aza-spiro [3.3] hept-6-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 14, step a). MS (m/e): 415(M + H).

Example 19

6- [ 2-tert-butyl-7- [ (2-methanesulfonylphenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane

In analogy to the procedure described for the synthesis of 6- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane (example 14) the title compound was prepared from 2-tert-butyl-4- (2-oxa-6-aza-spiro [3.3] hept-6-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 14, step a). MS (m/e): 441(M + H).

Example 20

6- [ 2-tert-butyl-7- [ (2-chloropyridin-3-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane

In analogy to the procedure described for the synthesis of 6- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane (example 14) the title compound was prepared from 2-tert-butyl-4- (2-oxa-6-aza-spiro [3.3] hept-6-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 14, step a). MS (m/e): 398(M + H).

Example 21

6- [ 2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane

In analogy to the procedure described for the synthesis of 6- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane (example 14) the title compound was prepared from 2-tert-butyl-4- (2-oxa-6-aza-spiro [3.3] hept-6-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 14, step a). MS (m/e): 430.8(M + H).

Example 22

6- [ 2-tert-butyl-7- [ (4-methyl-1, 2, 5-)Oxadiazol-3-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]-2-oxa-6-azaspiro [3.3]Heptane (Heptane)

Example 23

6- [ 2-tert-butyl-7- [ (5-methyl-1, 3, 4-Oxadiazol-2-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]-2-oxa-6-azaspiro [3.3]Heptane (Heptane)

In analogy to the procedure described for the synthesis of 6- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane (example 14) the title compound was prepared from 2-tert-butyl-4- (2-oxa-6-aza-spiro [3.3] hept-6-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 14, step a). MS (m/e): 369.6(M + H).

Example 24

1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

a) Acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester

The title compound was prepared in analogy to the procedure described for the synthesis of 2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 1, step c) from 2-tert-butyl-4-chloro-7H-pyrrolo [2, 3-d ] pyrimidine and pyrrolidin-3-yl acetate. MS (m/e): 303(M + H).

b)1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

To acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] at 0 deg.C]To a solution of pyrimidin-4-yl) -pyrrolidin-3-yl ester (100mg, 0.33mmol) in dry DMF (5mL) was added NaH (60% in oil; 40 mg; 0.99mmol) and the reaction mixture was stirred at 25 ℃ for 1 h. 1-bromomethyl-2-chloro-benzene (135.3mg, 0.66mmol) was added at 0 ℃ and the reaction mixture was stirred at 25 ℃ for 12 h. The mixture is treated with NH₄Aqueous Cl (10mL) was quenched and extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with water (10mL) and over anhydrous Na₂SO₄Dried, filtered and evaporated. By passingPreparative HPLC [ Xterra-RP18, 10. mu.19X 250 mM/acetonitrile/10 mM, using ammonium acetate in water as solvent system]The residue was purified to give the title compound (60mg, 46%) as a colorless viscous solid. MS (m/e): 385(M + H).

Example 25

1- [ 2-tert-butyl-7- [ (2-chloro-4-fluorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

The title compound was prepared in analogy to the procedure described for the synthesis of 1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 24) from acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester (example 24, step a). MS (m/e): 403(M + H).

Example 26

1- [ 2-tert-butyl-7- [ (2-methanesulfonylphenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

The title compound was prepared in analogy to the procedure described for the synthesis of 1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 24) from acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester (example 24, step a). MS (m/e): 429(M + H).

Example 27

1- [ 2-tert-butyl-7- [ [2- (trifluoromethyl) phenyl ] methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

The title compound was prepared in analogy to the procedure described for the synthesis of 1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 24) from acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester (example 24, step a). MS (m/e): 419(M + H).

Example 28

1- [ 2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

Example 29

1- [ 2-tert-butyl-7- [ (2-chloropyridin-3-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

The title compound was prepared in analogy to the procedure described for the synthesis of 1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 24) from acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester (example 24, step a). MS (m/e): 386(M + H).

Example 30

1- [ 2-tert-butyl-7- [ (3-chloropyridin-2-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

Example 31

1- [ 2-tert-butyl-7- [ (3-methyl-1, 2, 4-Oxadiazol-5-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-ols

The title compound was prepared in analogy to the procedure described for the synthesis of 1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 24) from acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester (example 24, step a). MS (m/e): 357(M + H).

Example 32

1- [ 2-tert-butyl-7- [ (4-methyl-1, 2, 5-)Oxadiazol-3-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-ols

Example 33

1- [ 2-tert-butyl-7- [ (5-methyl-1, 3, 4-Oxadiazol-2-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-ols

Example 34

1- [ 2-tert-butyl-7- [ (1-methyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

Example 35

Acetic acid [1- [ 2-tert-butyl-7- [ (1-methyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-yl ] ester

To acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] at 0 deg.C]Pyrimidin-4-yl) -pyrrolidin-3-yl ester (example 24, step a) (50mg, 0.16mmol) in dry DMF (5mL) was added NaH (60% in oil; 10 mg; 0.192mmol) and the reaction mixture was stirred at 25 ℃ for 1 h. 5-chloromethyl-1-methyl-1H-tetrazole (28mg, 0.20mmol) was then added to this solution at 0 ℃ and the reaction mixture was stirred at 25 ℃ for 12H. The mixture is treated with NH₄Aqueous Cl (10mL) was quenched and extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with water (10mL) and over anhydrous Na₂SO₄Dried, filtered and evaporated. By preparative HPLC [ Xterra-RP18, 10. mu.19X 250 mM/acetonitrile/10 mM, using ammonium acetate in water as solvent system]The residue was purified to give the title compound as an off-white solid (32mg, 48%). MS (m/e): 399.2(M + H).

Example 36

Acetic acid [1- [ 2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-yl ] ester

In analogy to the procedure described for the synthesis of [1- [ 2-tert-butyl-7- [ (1-methyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-yl ] acetate (example 35) the title compounds acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester and 1-bromomethyl-2, 3-dichloro-benzene were prepared. MS (m/e): 460.8(M + H).

Example 37

Acetic acid [1- [ 2-tert-butyl-7- [ (4-methyl-1, 2, 5-)Oxadiazol-3-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-yl radical]Esters

In analogy to the procedure described for the synthesis of [1- [ 2-tert-butyl-7- [ (1-methyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-yl ] acetate (example 35) the title compound was prepared from acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester and 3-chloromethyl-4-methyl-furazan. MS (m/e): 399(M + H).

Example 38

1- [ 2-tert-butyl-7- [ (1-cyclopropyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

The title compound was prepared in analogy to the procedure described for the synthesis of 1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 24) from acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester (example 24, step a). MS (m/e): 383(M + H).

Example 39

1- [ 2-tert-butyl-7- [ (3R) -tetrahydrofuran-3-yl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

The title compound was prepared in analogy to the procedure described for the synthesis of 1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 24) from acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester (example 24, step a). MS (m/e): 331.2(M + H).

Example 40

1- [ 2-tert-butyl-7- [ [3- (trifluoromethyl) -1H-pyrazol-4-yl ] methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

The title compound was prepared in analogy to the procedure described for the synthesis of 1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 24) from acetic acid 1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl ester (example 24, step a). MS (m/e): 409.2(M + H).

EXAMPLE 41

2-tert-butyl-4- (3, 3-difluoropyrrolidin-1-yl) -7- [ [3- (trifluoromethyl) -1H-pyrazol-4-yl ] methyl ] pyrrolo [2, 3-d ] pyrimidine

In analogy to the procedure described for the synthesis of 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] -4- (3, 3-difluoropyrrolidin-1-yl) pyrrolo [2, 3-d ] pyrimidine (example 1) the title compound was prepared from 2-tert-butyl-4- (3, 3-difluoro-pyrrolidin-1-yl) -7H-pyrrolo [2, 3-d ] pyrimidine (example 1, step c). MS (m/e): 428.8(M + H).

Example 42

(3S) -1- [ 2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

Preparative HPLC on reprosil Chiral NR of 1- [ 2-tert-butyl-7- [ (2, 3-dichlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 28) gave the title compound as a white foam. MS (m/e): 419.1(M + H).

Example 43

(3S) -1- [ 2-tert-butyl-7- [ (2-chloro-4-fluorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

Preparative HPLC on reprosil Chiral NR of 1- [ 2-tert-butyl-7- [ (2-chloro-4-fluorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 25) gave the title compound as a colourless foam. MS (m/e): 403.1(M + H).

Example 44

(3S) -1- [ 2-tert-butyl-7- [ [2- (trifluoromethyl) phenyl ] methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

Preparative HPLC isolation of 1- [ 2-tert-butyl-7- [ [2- (trifluoromethyl) phenyl ] methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 27) on reprosil Chiral Chiral NR gave the title compound as a white foam. MS (m/e): 419.2(M + H).

Example 45

(3S) -1- [ 2-tert-butyl-7- [ (2-methanesulfonylphenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

Preparative HPLC on reprosil Chiral NR of 1- [ 2-tert-butyl-7- [ (2-methanesulfonylphenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 26) gave the title compound as a white foam. MS (m/e): 429.2(M + H).

Example 46

(3S) -1- [ 2-tert-butyl-7- [ (2-chloropyridin-3-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

a) (S) -1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl acetate

b) (3S) -1- [ 2-tert-butyl-7- [ (2-chloropyridin-3-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

The title compound was prepared in analogy to the procedure described for the synthesis of 1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 24) from (S) -1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl acetate (example 46, step a). MS (m/e): 386(M + H).

Example 47

(3S) -1- [ 2-tert-butyl-7- [ (3-chloropyridin-2-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol

Example 48

(3S) -1- [ 2-tert-butyl-7- [ (4-methyl-1, 2, 5-)Oxadiazol-3-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]Pyrrolidin-3-ols

The title compound was prepared in analogy to the procedure described for the synthesis of 1- [ 2-tert-butyl-7- [ (2-chlorophenyl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] pyrrolidin-3-ol (example 24) from (S) -1- (2-tert-butyl-7H-pyrrolo [2, 3-d ] pyrimidin-4-yl) -pyrrolidin-3-yl acetate (example 46, step a). MS (m/e): 357.2(M + H).

Example 49

6- [ 2-tert-butyl-7- [ (3-chloropyridin-2-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane

Example 50

6- [ 2-tert-butyl-7- [ (1-methyltetrazol-5-yl) methyl ] pyrrolo [2, 3-d ] pyrimidin-4-yl ] -2-oxa-6-azaspiro [3.3] heptane

Example 51

Pharmacological testing

The following assays were performed to determine the activity of the compounds of formula I:

radioligand binding assays

Affinity of the compounds of the invention for cannabinoid CB1 receptor use suggested amounts of membrane preparations (PerkinElmer) of Human Embryonic Kidney (HEK) cells expressing the human CNR1 or CNR2 receptor each bound 1.5 or 2.6nM [ 3H) respectively]-CP-55, 940(Perkin Elmer) was determined as radioligand. Binding to a total volume of 0.2ml of binding buffer (50 mM Tris, 5mM MgCl2, 2.5mM EDTA for CB1 receptor, and 0.5% (wt/vol) fatty acid free BSA, pH 7.4, and 50mM Tris, 5mM MgCl for CB2 receptor, pH 7.4₂2.5mM EGTA, and 0.1% (wt/vol) fatty acid free BSA, pH 7.4), shaking at 30 ℃ for 1 h. Through a microfilter plate (UniFilter GF/B) coated with 0.5% polyethyleneimineA filter plate; packard) the reaction was terminated by rapid filtration. Bound radioactivity was analyzed using non-linear regression analysis (Activity Base, ID Businesssolution, Limited) for Ki for [3H]Kd values for CP55, 940 were determined from saturation experiments. The compounds of formula (I) show excellent affinity for the CB2 receptor, below 10 μ M, more particularly 1nM to 3 μ M and most particularly 1nM to 100 nM.

cAMP assay

CHO cells expressing human CB1 or CB2 receptors were seeded 17-24 hours prior to the experiment at 50.000 cells/well in black 96-well plates with clear flat bottom (Corning Costar #3904), in DMEM (Invitrogen No.31331), supplemented with 1 HT, with 10% fetal bovine serum, and in a humidified incubator at 5% CO₂And incubated at 37 ℃. The medium was exchanged with Krebs Ringer Bicarbonate buffer with 1mM IBMX and incubated at 30 ℃ for 30 minutes. The compound was added to a final assay volume of 100. mu.l and incubated at 30 ℃ for 30 minutes. Using the cAMP-Nano-TRF detection kit (Roche Diagnostics), by adding 50. mu.l lysis reagent (Tris, NaCl, 1.5% Triton X100, 2.5% NP40, 10% NaN)₃) And 50. mu.l of detection solution (20. mu.M mAb Alexa700-cAMP 1: 1, and 48. mu.M ruthenium-2-AHA-cAMP) stop the assay and oscillate at room temperature for 2 h. By being equipped with ND: a TRF reader (evotech technologies GmbH) with YAG laser as excitation source measures the time-resolved energy transfer. The plate was measured twice, excited at 355nm and emitted at 730 (bandwidth 30nm) or 645nm (bandwidth 75nm) with a 100ns delay and a 100ns gate (gate) respectively, for a total exposure time of 10 s. The FRET signal is calculated as follows: FRET-T730-Alexa 730-P (T645-B645), P-Ru 730-B730/Ru645-B645, where T730 is the test well measured at 730nM, T645 is the test well measured at 645nM, and B730 and B645 are buffer controls at 730nM and 645nM, respectively. cAMP content is determined from a function spanning a standard curve from 10. mu.M to 0.13nM cAMP.

EC is determined using Activity Base analysis (ID Business Solution, Limited)₅₀The value is obtained. EC for a broad range of cannabinoid agonists generated from this assay₅₀Value and scientific literatureThe disclosed values are matched.

The compounds of the present invention are CB2 receptor agonists, the EC thereof₅₀Less than 1 μ M and at least 10-fold selective in the corresponding assay relative to CB 1. Particular compounds of the invention are CB2 receptor agonists, the EC thereof₅₀Less than 0.05 μ M and at least 500-fold selective in the corresponding assay relative to CB 1.

For example, the following compounds show the following human EC in the above-described functional cAMP assay₅₀The value:

example A

Film-coated tablets containing the following ingredients can be prepared in a conventional manner:

composition (I)	Each sheet is
			And (3) nucleus:
a compound of formula (I)	10.0mg	200.0mg
			Microcrystalline cellulose	23.5mg	43.5mg
Hydrous lactose	60.0mg	70.0mg
			Povidone (Povidone) K30	12.5mg	15.0mg
Sodium starch glycolate	12.5mg	17.0mg
			Magnesium stearate	1.5mg	4.5mg
(nuclear weight)	120.0mg	350.0mg
			Film coating:
hydroxypropyl methylcellulose	3.5mg	7.0mg
			Polyethylene glycol 6000	0.8mg	1.6mg
Talc	1.3mg	2.6mg
			Iron oxide (yellow)	0.8mg	1.6mg
Titanium dioxide	0.8mg	1.6mg

The active ingredient is sieved and mixed with microcrystalline cellulose and the mixture is granulated with an aqueous solution of polyvinylpyrrolidone. The granules were then mixed with sodium starch glycolate and magnesium stearate and compressed to obtain 120 or 350mg of cores, respectively. The core is coated with the aqueous solution/suspension of the film coating described above.

Example B

Capsules containing the following ingredients can be prepared in a conventional manner:

composition (I)	Each sheet is
		A compound of formula (I)	25.0mg
Lactose	150.0mg
		Corn starch	20.0mg
Talc	5.0mg

The ingredients were sieved and mixed and filled into size 2 capsules.

Example C

The injection solution may have the following composition:

a compound of formula (I)	3.0mg
		Polyethylene glycol 400	150.0mg
Acetic acid	In a proper amount to obtain pH 5.0
		Water for injection	Adding to 1.0ml

The active ingredient is dissolved in a mixture of polyethylene glycol 400 and water for injection (a portion). The pH was adjusted to 5.0 by adding acetic acid. The volume was adjusted to 1.0ml by adding the balance of water. The solution was filtered, filled into vials with the appropriate excess and sterilized.

Claims

1. A compound of formula (I):

wherein

A is CH₂Or is absent;

R¹and R²Together with the nitrogen atom to which they are attached form a substituted pyrrolidinyl or 2-oxa-6-azaspiro [3.3]Heptyl wherein the substituted pyrrolidinyl is substituted with one or two independently selected from haloElement, hydroxy and C_1-8Pyrrolidinyl substituted with a substituent in alkylcarbonyloxy; and is

R³Is halophenyl, C_1-8Alkylsulfonylphenyl, halo C_1-8Alkylphenyl, halopyridyl, C_1-8Alkyl radicalOxadiazolyl, C_1-8Alkyl triazolyl, C_1-8Alkyl tetrazolyl, tetrahydrofuryl, C_3-8Cycloalkyl tetrazolyl or halo C₁-₈alkyl-1H-pyrazolyl;

or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein A is CH₂。

3. The compound of claim 1 or 2, wherein R¹And R²Together with the nitrogen atom to which they are attached form a substituted pyrrolidinyl group, wherein substituted pyrrolidinyl is pyrrolidinyl substituted with one or two substituents independently selected from halogen and hydroxy.

4. The compound of claim 1 or 2, wherein R¹And R²Together with the nitrogen atom to which they are attached form a substituted pyrrolidinyl group, wherein substituted pyrrolidinyl is pyrrolidinyl substituted with one or two substituents independently selected from fluoro and hydroxy.

5. The compound of claim 1 or 2, wherein R¹And R²Together with the nitrogen atom to which they are attached form a difluoropyrrolidinyl or hydroxypyrrolidinyl group.

6. The compound of claim 1 or 2, wherein R³Is halophenyl, halo C_1-8Alkyl phenyl, C_1-8Alkylsulfonylphenyl, halopyridyl or C_1-8Alkyl radicalA diazolyl group.

7. The compound of claim 1 or 2, wherein R³Is dichlorophenyl, chlorophenyl, trifluoromethylphenyl, methylsulfonylphenyl, chloropyridyl or methylA diazolyl group.

8. A compound according to claim 1 or 2, selected from

6- [ 2-tert-butyl-7- [ (5-methyl-1, 3, 4-Oxadiazol-2-yl) methyl]Pyrrolo [2, 3-d]Pyrimidin-4-yl]-2-oxa-6-azaspiro [3.3]Heptane;

9. A compound according to claim 1 or 2, selected from

10. A process for the preparation of a compound according to any one of claims 1 to 9, which process comprises reacting a compound of formula (a) at X-a-R³Reaction in the presence of:

wherein X is a leaving group and wherein A and R¹To R³Is as defined in any one of claims 1 to 7.

11. A pharmaceutical composition comprising a compound according to any one of claims 1 to 9 and a therapeutically inert carrier.

12. Use of a compound according to any one of claims 1 to 9 for the preparation of a medicament for the treatment or prophylaxis of pain, atherosclerosis, age-related macular degeneration, diabetic retinopathy, glaucoma, retinal vein occlusion, retinopathy of prematurity, ocular ischemic syndrome, geographic atrophy, diabetes mellitus, inflammation, inflammatory bowel disease, ischemia-reperfusion injury, acute liver failure, liver fibrosis, lung fibrosis, kidney fibrosis, systemic fibrosis, acute allograft rejection, chronic allograft nephropathy, diabetic nephropathy, glomerulonephropathy, cardiomyopathy, heart failure, myocardial ischemia, myocardial infarction, systemic sclerosis, thermal injury, burning, hypertrophic scars, keloids, gingivitis pyrexia, liver cirrhosis or tumors, amyotrophic lateral sclerosis, multiple sclerosis, alzheimer's disease, parkinson's disease, stroke or transient ischemic attacks, or for bone regulation.

13. Use of a compound according to any one of claims 1 to 9 for the manufacture of a medicament for the treatment or prevention of uveitis.