HK1168600A - Novel p2x7r antagonists and their use - Google Patents

Description

Novel P2X7R antagonists and uses thereof

The present application relates to novel P2X7R antagonists of N-indol-3-yl-acetamide and N-azaindol-3-yl-acetamide compounds, pharmaceutical compositions comprising these compounds and their use in the prophylactic and therapeutic treatment of diseases or conditions mediated by P2X 7R.

Background

P2X7R is an ATP-gated ion channel belonging to the P2X ion transitional channel family. The gene was first isolated from rat brain (Surprenant et al (1996) 272: 735-738) and subsequently isolated from the human monocyte pool by virtue of its sequence homology to other members of the P2X family (Rassenden et al (1997) J.biol.chem.272: 5482-5486; Gene bank (Genbank) accession No. NM-002562, Y09561). P2X7R was later found to be comparable to the unidentified P2Z receptor that mediates ATP penetration of mast cells and macrophages (Dahlqvist and Diamant (1974) Acta Physiol.Scand.34: 368-. P2X7R has two hydrophobic transmembrane domains, an extracellular loop, and forms a transmembrane ion channel. P2X7R has pharmacological properties that are significantly different from those of other P2X homo-or heteromers (North and Surprenant (2000) Annual Rev. pharmacological Toxicology 40: 563-580). P2X7R requires ATP levels in excess of 1mM to effect activation, whereas the other P2X receptors are activated at ATP concentrations ≦ 100 μ M (Steinberg et al (1987) J.biol. chem.262: 8884-8888; Greenberg et al (1988) J.biol. chem.263: 10337-10343). Although all P2X receptors exhibit non-selective channel-like properties upon binding, the channel formed by P2X7R can be rapidly converted into a small pore that allows passage of molecules up to 900 daltons (Virginio et al (1999) J. Physiol.519: 335-346).

P2X7R is expressed in hematopoietic cells, mast cells, lymphocytes, erythrocytes, fibroblasts, Langerhans' cells and macrophages (Surprenant et al, 1996, Science 272: 3118-. In the central nervous system, it has been reported that P2X7R is expressed in glial cells, Schwann cells, astrocytes and in neurons (Ferrari et al (1996) J. Immunol 156: 1531-1539; Collo et al (1997) Neuropharmacology 36: 1277-1283; Anderson and Nedergard (2006) Trends Neuroscien 29: 257-262).

P2X7R is involved in the regulation of immune function and inflammatory responses. Activation of P2X7R by ATP in macrophages is associated with mitogenic stimulation of T cells (Baricordi et al (1996) Blood 87: 682-690), cytokine release (Griffiths et al (1995) J.Immol.154: 2821-2828) and formation of macrophage polykaryons (Falzoni et al (1995) J.Clin.invest.95: 1207-1216). P2X7R is involved in the processing and release of active interleukin-1 beta (IL-1 beta) in pro-inflammatory cells (Perregaux and Gabel (1998) J Biol Chem 269: 15195-. Stimulation of P2X7R by ATP also leads to apoptosis and Cell Death by triggering the formation of non-selective plasma membrane pores (Di Virgilio et al (1998) Cell Death Differ.5: 191-199).

Upregulation of P2X7R has been observed during ischemic injury and necrosis induced by occlusion of the middle cerebral arteries in rats (Collo et al (1997) Neuropharmacol 36: 1277-one 1283). Recent studies have indicated that P2X7R plays a role in the superoxide generation of microglia and that upregulation of P2X7R has been detected around amyloid plaques in transgenic mouse models for Alzheimer's disease (Parvathenani et al (2003) J Biol Chem 278: 13300-13317) and in multiple sclerosis lesions from post-mortem brain sections (Narcise et al (2005) Glia, 49: 245-258).

Studies in rats lacking inflammatory and neuropathic hypersensitivity to mechanical and thermal stimuli due to the lack of P2X7R indicate a correlation between P2X7R and inflammatory and neuropathic Pain (Chessell et al (2005) Pain 114: 386-396). Antagonists of P2X7R significantly improve functional recovery from spinal cord injury and reduce cell death in animal models (Wang et al (2004) Nature 10: B21-B27).

Compounds that modulate P2X7R have been reported. For example Brilliant Blue (Jiang et al, mol. Phamacol.58(2000), 82-88), isoquinoline 1- [ N, O-bis (5-isoquinolinesulfonyl) -N-methyl-L-tyrosyl ] -4-phenylpiperazine and N- [1- [ N-methyl-p- (5-isoquinolinesulfonyl) benzyl ] -2- (4-phenylpiperazine) ethyl ] -5-isoquinolinesulfonamide (Humphreys et al, mol. Pharmacol., 54(1998), 22-32), adamantane derivatives (WO 99/29660, WO 99/29661, WO 00/61569, WO01/42194, WO 01/44170, WO 01/44213, WO 01/94338, WO 03/041707, WO 03/042190, WO 03/080579), WO 04/074224, WO 05/014529, WO06/025783, WO 06/059945), piperidine and piperazine compounds (WO 01/44213, WO01/46200, WO 08/005368), benzamide and heteroaryl amide compounds (WO 03/042191, WO 04/058731, WO 04/058270, WO 04/099146, WO 05/019182, WO06/003500, WO 06/003513, WO 06/067444), substituted tyrosine derivatives (WO00/71529, WO 03/047515, WO 03/059353), imidazole compounds (WO 05/014555), amino-tetrazole compounds (WO 05/111003), cyanoamidines (WO 06/017406), bicyclic heteroaryl derivatives (WO 05/009968, WO 06/102588, WO 06/102610, WO 07/028022, WO 06/059945), WO 07/109154, WO 07/109160, WO 07/109172, WO 07/109182, WO07/109192, WO 07/109201), hydrazides (WO 06/110516) and other examples (WO99/29686, WO 04/106305, WO 05/039590, WO 06/080884, WO 06/086229, WO 06/136004, WO 07/025366, WO 07/056046, WO 07/056091, WO07/141267, WO 07/141269, WO 08/003697) are antagonists of P2X7R, while oxidized ATP (oATP) acts as an irreversible inhibitor of the receptor (Chen et al, J.biol.Chem., 268(1993), 8199 + 8203).

Thus, there is strong evidence that compounds that act on P2X7R are useful in the treatment of pain, inflammatory processes and degenerative conditions associated with disease states such as: rheumatoid arthritis, osteoarthritis, psoriasis, allergic dermatitis, asthma, chronic obstructive pulmonary disease, airway hyperreactivity, septic shock, glomerulonephritis, irritable bowel disease, inflammatory bowel disease, crohn's disease, ulcerative colitis, atherosclerosis, growth and metastasis of malignant cells, myoblastic leukemia, diabetes, alzheimer's disease, parkinson's disease, multiple sclerosis, glaucoma, age-related macular degeneration, uveitis, neuropathic pain, depression, bipolar disorder, anxiety, meningitis, traumatic brain injury, acute spinal cord injury, neuropathic pain, osteoporosis, burn injury, ischemic heart disease, myocardial infarction, stroke, and varicose veins.

It is therefore an object of the present invention to provide a series of novel compounds which inhibit P2X7R activity and which are useful in the treatment of the above mentioned diseases.

Detailed Description

The present invention relates to novel P2X7R antagonist N-indol-3-yl-acetamides and N-azaindol-3-yl-acetamides compounds represented by the general formula (I):

wherein

-R₁Is mono-or dicycloalkylalkyl or mono-or dicycloalkyl;

-R₂selected from straight or branched C₁-C₅Alkyl, which may be optionally substituted with:

-OH、-CH₂-OH、C₁-C₅alkoxy, NH₂-、N(R_a)₂-、NHR_a-、CN-、CF₃Halogen (i.e. Cl, F, Br or I), piperidin-1-yl, morpholin-4-yl, pyrrolidin-1-yl, 5H-tetrazolylpropyl, methylcarbamoyl, dimethylcarbamoyl or ethylmethylcarbamoyl, wherein R is_aIs C₁-C₅An alkyl group;

-R₃、R₄、R₅、R₆independently at each occurrence, selected from hydrogen, halogen (i.e., Cl, F, Br, or I), methyl, methoxy, cyano, or trifluoromethyl;

-a, b, c, d, x, at each occurrence, are independently selected from carbon or nitrogen; or a pharmaceutically acceptable salt or solvate thereof (wherein if x is carbon, it must have a hydrogen substituent).

Wherein R is₁Is selected from the group consisting of cyclopentyl, cyclopentylmethyl, cyclohexyl, cyclohexylmethyl, cycloHeptyl, cycloheptylmethyl, bicyclo [2.2.2]Octane-1-yl and bicyclo [2.2.2]Compounds of formula (I) of the group octane-1-ylmethyl are preferred.

Also preferred is where R₂A compound substituted with one or two substituents selected from the group consisting of: -OH, -CH₂-OH、C₁-C₅Alkoxy, -NH₂、NHRa、-CN、-CF₃Halogen, piperidin-1-yl, morpholin-4-yl, pyrrolidin-1-yl, or 5H-tetrazolylpropyl.

Wherein R is as disclosed above₂Is C₁-C₅Alkyl or C₂-C₅Hydroxyalkyl compounds are also preferred.

In addition, R is also preferred₃、R₄、R₅And R₆At least two of which are hydrogen. If a valence is desired, R₃-R₆May also be absent.

In addition, it is also preferred that a, b, C and d are C or that one of a, b, C and d is N.

Examples of novel N-indol-3-yl-acetamide and N-azaindol-3-yl-acetamide compounds are disclosed in examples 1-3.

The present invention further relates to compounds of formula (I) or a pharmaceutically acceptable salt or solvate thereof, which are:

-N- (4-chloro-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-chloro-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-chloro-1- (hydroxymethyl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-bromo-1- (hydroxymethyl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-chloro-1- (hydroxymethyl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-bromo-1- (hydroxymethyl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-chloro-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cyclohexylacetamide,

-N- (4-chloro-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide,

-N- (4-chloro-1- (2-hydroxypropyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (2-hydroxypropyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide,

-N- (4-chloro-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -3-cyclohexylpropionamide and

-N- (4-bromo-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -3-cyclohexylpropionamide.

-N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) acetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) acetamide,

-N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) acetamide

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) acetamide,

-N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -2- (bicyclo [2.2.2] octan-1-yl) acetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -2- (bicyclo [2.2.2] octan-1-yl) acetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-chloro-1-methyl-1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1-methyl-1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-chloro-1-methyl-1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1-methyl-1H-indol-3-yl) -2-cycloheptylacetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1-methyl-1H-indol-3-yl) acetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1-methyl-1H-indol-3-yl) acetamide,

-N- (4-chloro-1-methyl-1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-bromo-1-methyl-1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-chloro-1-methyl-1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-bromo-1-methyl-1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-chloro-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-chloro-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) acetamide, and

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) acetamide.

The invention also includes isotopically-labeled compounds, which are identical to those recited in formula (I), but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, respectively, e.g.²H、³H、¹³C、¹⁴C、¹⁵N、¹⁸O、¹⁷O、³¹P、³²P、³⁵S、¹⁸F and³⁵and (4) Cl. Containing the above isotopes and/or other isotopes of other atomsThe compounds of the present invention, prodrugs thereof, and pharmaceutically acceptable salts of said compounds or of said prodrugs are within the scope of the present invention. Certain isotopically-labelled compounds of the invention, e.g. in which a radioactive isotope is incorporated, e.g.³H and¹⁴the compounds of C are useful in drug and/or substrate tissue distribution assays. Tritium is³H and carbon-14 i.e¹⁴The C isotope is particularly preferred because of its ease of preparation and detection. In addition, with heavier isotopes such as deuterium²H substitution may result in certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and is therefore preferred in certain circumstances.

Isotopically labeled compounds of formula (I) and prodrugs thereof of the present invention are generally prepared by carrying out the procedures disclosed in the examples below by substituting a non-isotopically labeled reagent with a readily available isotopically labeled reagent.

Pharmaceutically acceptable salts include salts formed with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, and the like, and cations such as those derived from sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, ferric hydroxide, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine and procaine.

Other pharmaceutically acceptable salts

In a further embodiment, the present application relates to a pharmaceutical composition comprising a compound of formula (I) according to the present invention.

The pharmaceutical compositions of the present invention may also contain other active compounds in separate or unit dosage forms for simultaneous or sequential administration.

A compound of formula (I) or a pharmaceutically acceptable salt thereof, is useful in the manufacture of a medicament for the prophylactic or therapeutic treatment of any disease state in a human or other mammal that is exacerbated or caused by excessive or unregulated cytokine production by mammalian cells, such as, but not limited to, monocytes and/or macrophages.

The invention also relates to the treatment of IL-1 or cytokine mediated disorders.

"IL-1 mediated disorders" and "cytokine mediated disorders" as defined herein include, but are not limited to, diseases or disorders selected from the group consisting of mammals, including humans, from: arthritis (including psoriatic arthritis, Retle's syndrome, rheumatoid arthritis, gout, traumatic arthritis, rubella arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and acute synovitis), inflammatory bowel disease, Crohn's disease, emphysema, acute respiratory distress syndrome, adult respiratory distress syndrome, asthma, bronchitis, chronic obstructive pulmonary disease, chronic pulmonary inflammatory disease, silicosis, pulmonary sarcoidosis, anaphylaxis, allergic contact hypersensitivity, eczema, contact dermatitis, psoriasis, sunburn, cancer, tissue ulceration, restenosis, periodontal disease, epidermolysis bullosa, osteoporosis, bone resorption diseases, joint loss, atherosclerosis, aortic aneurysm, congestive heart failure, myocardial infarction, stroke, cerebral ischemia, head trauma, neurotrauma, neuro, rheumatoid arthritis, gout, rheumatoid arthritis, osteoarthritis, asthma, allergic contact hypersensitivity, eczema, contact dermatitis, psoriasis, sunburn, cancer, tissue ulceration, restenosis, periodontal disease, epidermolysi, Spinal cord injury, neurodegenerative disease, Alzheimer's disease, Parkinson's disease, glaucoma, age-related macular degeneration, uveitis, neuropathic pain, migraine, depression, peripheral neuropathy, pain, cerebellar amyloid angiopathy, mental or cognitive enhancement, amyotrophic lateral sclerosis, multiple sclerosis, ocular angiogenesis, corneal injury, macular degeneration, corneal scarring, scleritis, abnormal wound healing, burns, autoimmune disease, Huntington's chorea, diabetes, AIDS, cachexia, sepsis, septic shock, endotoxic shock, conjunctivitis inflammatory shock, gram-negative sepsis, toxic shock syndrome, cerebellar malaria, cardiac and renal reperfusion injury, thrombosis, glomerulonephritis, graft-versus-host reaction, allograft rejection, organ transplant toxicity, ulcerative colitis or muscle degeneration, comprising administering to said mammal an amount of a compound of formula (I) effective to treat the condition.

The present invention relates to a pharmaceutical composition for the treatment of IL-1 mediated disorders in mammals, including humans, comprising a compound of formula (I) in an amount effective to treat the disorder and a pharmaceutically acceptable carrier.

The compounds of the invention are useful in the treatment of rheumatoid arthritis, osteoarthritis, psoriasis, allergic dermatitis, asthma, Chronic Obstructive Pulmonary Disease (COPD), airway hyperresponsiveness, septic shock, glomerulonephritis, irritable bowel disease, crohn's disease, ulcerative colitis, atherosclerosis, growth and metastasis of malignant cells, myoblastic leukemia, diabetes, alzheimer's disease, meningitis, osteoporosis, burns, ischemic heart disease, stroke, and varicose veins.

In another aspect, the present invention also provides a pharmaceutical composition for use in the treatment of osteoarthritis, the pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) as defined above, or a pharmaceutically acceptable salt or solvate thereof.

The invention also provides a pharmaceutical composition for effecting immunosuppression, for example for the treatment of rheumatoid arthritis, irritable bowel disease, atherosclerosis or psoriasis, which comprises a therapeutically effective amount of a compound of formula (I) as hereinbefore defined or a pharmaceutically acceptable salt or solvate thereof.

The present invention also provides a pharmaceutical composition for use in the treatment of an obstructive airways disease, such as asthma or COPD, which comprises a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined.

The invention also provides pharmaceutical compositions for treating mammals susceptible to or suffering from conditions etiologically associated with aberrant activity of the P2X7 receptor, such as neurodegenerative diseases and conditions including, for example, parkinson's disease, multiple sclerosis, glaucoma, age-related macular degeneration, uveitis, neuropathic pain, diseases and conditions mediated or caused by neuroinflammation such as traumatic brain injury and encephalitis; centrally mediated neuropsychiatric diseases and conditions such as depression, mania, bipolar disorder, anxiety, schizophrenia, eating disorders, sleep disorders and cognitive disorders, epilepsy and seizures, which pharmaceutical composition comprises a therapeutically effective amount of a compound of formula (I) as defined above or a pharmaceutically acceptable salt or solvate thereof.

In particular embodiments, the pharmaceutical compositions of the present invention are useful for treating affective disorders. In a preferred embodiment, the affective disorder is selected from depression, anxiety, bipolar disorder and schizophrenia.

In another embodiment, the pharmaceutical compositions of the invention are useful for treating neurodegenerative diseases and disorders, diseases and disorders mediated by or causing neuroinflammation, and centrally-mediated neuropsychiatric diseases and disorders.

Furthermore, the pharmaceutical compositions of the invention are also particularly useful for the treatment of pain, inflammatory processes and degenerative disorders. In a more preferred embodiment, the inflammatory process is selected from the group consisting of rheumatoid arthritis, osteoporosis and chronic obstructive pulmonary disease.

In addition, the pharmaceutical compositions of the present invention are also useful for the treatment of neuropathic pain.

The dosages, Pharmaceutical formulations and delivery Forms of the compounds of formula (I) for use in accordance with the present invention may be formulated in conventional manner according to methods known in the art using one or more physiological carriers or excipients, see, e.g., Ansel et al, "Pharmaceutical Dosage Forms and drug delivery Systems", 7 th edition, Lippincott Williams & Wilkins Publishers, 1999. Thus, the P2X7R modulators and physiologically acceptable salts and solvates thereof are formulated for administration by inhalation, insufflation (either through the mouth or nose), oral, buccal, parenteral or rectal administration.

For oral administration, the pharmaceutical compositions of the compounds of formula (I) may be in the form of tablets or capsules prepared, for example, by conventional methods with pharmaceutically acceptable excipients such as binding agents (e.g., pregelatinized corn starch, polyvinylpyrrolidone, hydroxypropylmethylcellulose), fillers (e.g., lactose, microcrystalline cellulose, dibasic calcium phosphate), lubricants (e.g., magnesium stearate, talc, silicon dioxide), disintegrants (e.g., potato starch, sodium starch glycolate) or wetting agents (e.g., sodium lauryl sulfate). The pharmaceutical composition may be administered to a patient with a physiologically acceptable carrier, as described herein. In a particular embodiment, the term "pharmaceutically acceptable" means permitted by regulatory agencies or other generally recognized pharmacopoeias for use in animals, and more particularly in humans. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which a therapeutic agent is administered. The pharmaceutically acceptable carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium ions, dried skim milk, glycerol, propylene glycol, water, ethanol and the like. The composition may also contain minor amounts of wetting or emulsifying agents or pH buffering agents, if desired. These compositions may be in the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained release formulations, and the like. The composition may be formulated in the form of suppositories with conventional binders and carriers such as triglycerides. Oral formulations include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and the like. Examples of suitable Pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by e.w. martin. The compositions contain a therapeutically effective amount of the above-mentioned compounds, preferably in pure form, and a suitable amount of a carrier to provide the appropriate form of administration to the patient. The formulation should be adapted to the mode of administration.

Liquid preparations for oral administration may be in the form of, for example, solutions, syrups or suspensions, or may be presented as a dry preparation for constitution with water or other suitable vehicle before use. The liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g. sorbitol, syrup, cellulose derivatives, hydrogenated edible fats), emulsifying agents (e.g. lecithin, acacia), non-aqueous vehicles (e.g. almond oil, oily esters, ethyl alcohol, fractionated vegetable oils), preservatives (e.g. methyl or propyl p-hydroxybenzoates, sorbic acid). The preparation may also contain appropriate amount of buffer salt, correctant, colorant and sweetener. Formulations for oral administration may suitably be formulated to provide controlled release of the compound of formula (I).

For administration by inhalation, the compounds of formula (I) of the present invention are conveniently delivered in the form of an aerosol formulation, given from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of a compound of formula (I) and a suitable powder base such as lactose or starch.

The compounds of formula (I) of the present invention may be formulated for parenteral administration by injection, for example by bolus injection or continuous infusion. The site of injection includes intravenous, intraperitoneal or subcutaneous. Formulations for injection may be presented in unit dosage form (e.g., in vials, in multi-dose containers) with an added preservative. The compounds of formula (I) according to the invention may be in the form of suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing or dispersing agents. Alternatively, the active agent may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. Where desired, the composition may also include a solubilizing agent and a local anesthetic such as lignocaine to reduce pain at the site of injection. The ingredients are typically provided separately or mixed together in unit dosage form, for example in the form of a dry lyophilized powder or anhydrous concentrate in an air tight container such as an ampoule or sachet indicating the amount of active agent. In the case of administration of the composition by infusion, it is dispersed in an infusion bottle containing sterile pharmaceutical grade water or saline. In the case of administration of the composition by injection, an ampule of sterile water for injection or saline may be provided so that the components are mixed prior to administration.

The compounds of formula (I) of the present invention may be formulated for transdermal administration. Transdermal compositions are typically formulated in the form of a topical ointment or cream, which typically contains the active ingredient in an amount of from about 0.01 to about 20% by weight, preferably from about 0.1 to about 10% by weight, more preferably from about 0.5 to about 15% by weight. When formulated as an ointment, the active ingredient is typically mixed with an ointment base that is miscible with paraffin or miscible with water. Alternatively, the active ingredient may be formulated in a cream, for example, with an oil-in-water cream base. Such transdermal formulations are known in the art and typically include other ingredients to enhance the stable transdermal properties of the active ingredient or formulation. All such known transdermal formulations and ingredients are included within the scope of the present invention. The compounds of the present invention may also be administered by transdermal means. Thus, transdermal administration can be accomplished using a patch of the reservoir or porous membrane type or a patch of the solid matrix type.

The pharmaceutical composition of the present invention may be formulated in a neutral or salt form. Pharmaceutically acceptable salts include salts with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, and the like, and salts with cations such as those derived from sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, ferric hydroxide, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, and the like.

If desired, the compounds of formula (I) according to the invention may also be in the form of a pack or dispenser device which may contain one or more unit dosage forms containing the active agent. The package may for example comprise a metal or plastic sheet, such as a blister pack. The package or dispenser device may be accompanied by instructions for administration.

The compounds of formula (I) of the present invention may be administered as the sole active ingredient or in combination with other active agents. These active agents include non-steroidal anti-inflammatory drugs (NSAIDS), such as celecoxib, rofecoxib, cimicic, etoricoxib, lumiracoxib, valdecoxib, deracoxib, N- (2-cyclohexyloxynitrophenyl) methanesulfonamide, COX189, ABT963, JTE-522, GW-406381, LAS-34475, CS-706, PAC-10649, SVT-2016, GW-644784, tenidap (tenidap), acetylsalicylic acid (aspirin), Amoxiprin, benorilate, choline magnesium salicylate, diflunisal, Faislamine, methyl salicylate, magnesium salicylate, salsalate (salsalate), diclofenac, aceclofenac, acemetacin, bromfenac, etodolac, indomethacin, nabumetone, sulindac, tolmetin, ibuprofen, carprofen, ibuprofen, carprofen, flurbiprofen, ibuprofen, and ibuprofen, Ketorolac, loxoprofen, naproxen, oxaprozin, tiaprofenic acid, suprofen, mefenamic acid, meclofenamic acid, phenylbutazone, azapropazone, analgin, oxyphenbutazone, fensulazone, piroxicam, lornoxicam, meloxicam, tenoxicam, nimesulide, lincomone, paracetamol.

The compounds of formula (I) of the invention may be administered with active agents such as TNF-alpha inhibitors such as anti-TNF monoclonal antibodies (such as Remicade, CDP-870 and D2E7) and TNF receptor immunoglobulin molecules (such as Enbrel), low dose methotrexate, leflunomide; ciclesonide; hydroxychloroquine, d-penicillamine, auranofin or a combination of parenteral or oral gold.

The compounds of formula (I) according to the invention can also be used in combination with inhibitors of proTNF alpha-converting enzyme (TACE) such as 3-amino-N-hydroxy-alpha- (2-methylpropyl) -3- [4- [ (2-methyl-4-quinolyl) methoxy ] phenyl ] -2-oxo-1-pyrrolidineacetamide, 2(S), 3(S) -piperidinedicarboxamide, N3-hydroxy-1-methyl-N-2- [4- [ (2-methyl-4-quinolyl) methoxy ] phenyl ], 3-thiomorpholinecarboxamide, 4- [ [4- (2-butynyloxy) phenyl ] sulfonyl ] -N-hydroxy-2, 2-dimethylmorpholinecarboxamide, N-hydroxy, 5-hexenoic acid, 3- [ (hydroxyamino) carbonyl ] -2- (2-methylpropyl) -6-phenyl-2- (2-methylpropyl) -2- (methylsulfonyl) hydrazide, (2R, 3S, 5E), 2-piperidinecarboxamide, N, 5-dihydroxy-1- [ [4- (1-naphthylmethoxy) phenyl ] sulfonyl ] -, (2R, 5R), pentanamide, 3- (formylhydroxyamino) -4-methyl-2- (2-methylpropyl) -N- [ (1S, 2S) -2-methyl-1- [ (2-pyridinylamino) carbonyl ] butyl ] - (2R, 3S), 2-acrylamide, N-hydroxy-3- [3- [ [ (4-methoxyphenyl) sulfonyl ] (1-methylethyl) amino ] phenyl ] -3- (3-pyridyl) -, (2E), benzamide, N- (2, 4-dioxo-1, 3, 7-triazaspiro [4.4] non-9-yl) -4- [ (2-methyl-4-quinolyl) methoxy ], benzamide, N- [ (1-acetyl-4-piperidyl) (2, 5-dioxo-4-imidazolidinyl) methyl ] -4- [ (2-methyl-4-quinolyl) methoxy ] or 2, 4-imidazolidinedione, or a pharmaceutically acceptable salt thereof, or a pharmaceutically, 5-methyl-5- [ [ [4- [ (2-methyl-4-quinolyl) methoxy ] phenyl ] sulfonyl ] methyl ] is co-administered. Further examples of TACE inhibitors are described in WO 99/18074, WO99/65867, US 6,225,311, WO 00/00465, WO 00/09485, WO 98/38179, WO 02/18326, WO 02/096426, WO 03/079986, WO 03/055856, WO03/053941, WO 03/040103, WO 03/031431, WO 03/024899, WO 03/016248, WO 04/096206, WO 04/033632, WO 04/108086, WO 04/043349, WO04/032846, WO 04/012663, WO 04/006925, WO 07/016597.

The compounds of formula (I) of the present invention may also be administered in combination with corticosteroids such as budesonide, corticosterone, cortisol, cortisone acetate, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, beclomethasone, fludrocortisone acetate, deoxycorticosterone acetate (doca), aldosterone.

The compounds of formula (I) of the invention may also be administered in combination with a β 2-adrenoceptor agonist such as formoterol, salbutamol (albuterol), levosalbutamol, terbutaline, pirbuterol, procaterol, metaproterenol, fenoterol, bitolterol mesylate, salmeterol, bambuterol, clenbuterol.

The compounds of formula (I) of the present invention may also be administered in combination with antidepressants such as sertraline, escitalopram, fluoxetine, bupropion, paroxetine, venlafaxine, trazodone, amitriptyline, citalopram, duloxetine, mirtazapine, nortriptyline, imipramine, lithium salts.

The compounds of formula (I) of the present invention may also be administered in combination with an antipsychotic such as chlorpromazine, fluphenazine, oxyperchlorperazine, prochlorperazine, thioridazine, trifluralazine, mesoridazine, promazine, trifluoroperazine, levomepromazine, promethazine, chlorprothixene, flupentixol, thiothixene, zucotinol, haloperidol, droperidol, pimozide, mepilone, benperidol, trifluperidol, clozapine, olanzapine, risperidone, quetiapine, ziprasidone, amisulpride, paliperidone, Bifeprunox, aripiprazole.

The compounds of formula (I) of the present invention may also be administered with leukotriene biosynthesis inhibitors, 5-lipoxygenase (5-LO) inhibitors or 5-lipoxygenase activating protein (FLAP) antagonists, such as zileuton; ABT-761; fenton; teposalin; a pipe of the tobacco; VIA-2291; etalocib; ketoprofen, Abt-79175; abt-85761; n- (5-substituted) thiophen-2-alkylsulfonamides; TDT-070; (ii) a lincomron; PEP-03; tenocic; 2, 6-di-tert-butylphenol hydrazone; methoxytetrahydropyrans such as Zeneca ZD-2138; compound SB-210661; pyridyl-substituted 2-cyanonaphthalene compounds such as L-739-010; 2-cyanoquinoline compounds such as L-746-530; indole and quinoline compounds such as MK-591, MK-886 and BAY x 1005 are administered in combination.

The compounds of formula (I) of the present invention may be used in combination with receptor antagonists for leukotriene LTB4, LTC4, LTD4 and LTE, for example phenothiazin-3-ones such as L-651, 392; amidino compounds such as CGS-25019 c; benzoxaamines such as Ontezolast; benzamidines such as BIIL 284/260; and compounds such as zafirlukast, arlukast, montelukast, pranlukast, vilelukast (MK-679), RG-12525, Ro-245913, iralukast (CGP 45715A), and BAY x 7195; the administration of montelukast is combined.

The compounds of formula (I) of the present invention may also be administered in combination with PDE4 inhibitors, including inhibitors of the isoform PDE 4D.

The compounds of formula (I) of the present invention may also be administered in combination with antihistamine H1 receptor antagonists including cetirizine, loratadine, desloratadine, fexofenadine, esmin, azelastine, and chlorpheniramine.

The compounds of formula (I) according to the invention can also be used in combination with gastroprotective H₂The receptor antagonist is administered in combination.

The compounds of formula (I) of the present invention may also be administered in combination with vasoconstrictive sympathomimetics of the alpha 1-and alpha 2-adrenergic receptor agonists, including propylhexedrine, phenylephrine, phenylpropanolamine, pseudoephedrine, naphazoline hydrochloride, oxymetazoline hydrochloride, tetrahydrozoline hydrochloride, xylometazoline hydrochloride, and ethylnorepinephrine hydrochloride.

The compounds of formula (I) of the present invention may be administered with anticholinergic agents including iprodione; tiotropium bromide; oxitropium bromide; pirenzepine; and tirezrin in combination. The invention furthermore relates to compounds of the invention and beta₁-to beta₄-adrenergic receptor agonists including metaproterenol, isoproterenol, salbutamol, formoterol, salmeterol, terbutaline, metaproterenol, bitolterol mesylate (bitolterol mesylate) and pirbuterol; or methylxanthines, including theophylline and aminophylline; sodium cromoglycate; or a combination of muscarinic receptor (M1, M2, and M3) antagonists.

The compounds of formula (I) of the invention may be administered in combination with a type I insulin-like growth factor (IGF-1) mimetic.

The compounds of formula (I) of the present invention may be administered in combination with inhaled glucocorticoids with reduced systemic side effects, including prednisone, prednisolone, flunisolide, triamcinolone acetonide, beclomethasone dipropionate, budesonide, fluticasone propionate and mometasone furoate.

The compounds of formula (I) of the present invention may be used with (a) tryptase inhibitors; (b) platelet Activating Factor (PAF) antagonists; (c) interleukin-converting enzyme (I), (II)ICE) inhibitors; (d) an IMPDH inhibitor; (e) adhesion molecule inhibitors, including VLA-4 antagonists; (f) (ii) a cathepsin; (g) MAP kinase inhibitors; (h) glucose-6 phosphate dehydrogenase inhibitors; (i) kinin-B₁-and B₂-a receptor antagonist; j) anti-gout agents, such as colchicine; (k) xanthine oxidase inhibitors such as allopurinol; (l) Uricosuric agents such as probenecid, sulindac, and benzbromarone; (m) growth hormone secretagogues; (n) transforming growth factor (TGF β); (o) Platelet Derived Growth Factor (PDGF); (p) fibroblast growth factor, such as basic fibroblast growth factor (bFGF); (q) granulocyte macrophage colony stimulating factor (GM-CSF); (r) capsaicin cream; (s) tachykinin NK₁And NK₃Receptor antagonists such as NKP-608C; SB-233412 (talnetant); and D-4418; and (t) elastase inhibitors, such as UT-77 and ZD-0892.

The compounds of formula (I) of the present invention can be used with Matrix Metalloproteinases (MMPs), i.e. stromelysin, collagenase and gelatinase, and proteoglycanase; in particular collagenase-1 (MMP-1), collagenase-2 (MMP-8), collagenase-3 (MMP-13), stromelysin-1 (MMP-3), stromelysin-2 (MMP-10) and stromelysin-3 (MMP-11) inhibitors.

The compounds of formula (I) of the present invention may be administered in combination with anti-cancer agents such as endostatin and angiostatin or cytotoxic drugs such as doxorubicin, daunomycin, cisplatin, etoposide, paclitaxel, taxotere and farnesyl transferase inhibitors, VEGF inhibitors, COX-2 inhibitors and anti-metabolites such as methotrexate, anti-tumour agents, particularly anti-mitotic agents, including vinca alkaloids such as vinblastine and vincristine.

The compounds of formula (I) of the present invention may be administered in combination with antiviral agents such as Viracept, AZT, acyclovir and famciclovir, and antibacterial compounds such as Valant.

The compounds of formula (I) of the present invention may be administered in combination with cardiovascular agents such as calcium channel blockers, lipid lowering agents such as statins, fibrates, beta-blockers, ACE inhibitors, angiotensin-2 receptor antagonists and platelet aggregation inhibitors.

The compounds of formula (I) of the present invention may be administered in combination with CNS agents such as antidepressants (such as sertraline), anti-Parkinson agents (such as Deprenyl, L-dopa, Requip, Mirapex, MAOB inhibitors such as selegiline and rasagiline, comp inhibitors such as Tasmar, A-2 inhibitors, dopamine reuptake inhibitors, NMDA antagonists, nicotine agonists, dopamine agonists and inhibitors of neuronal nitric oxide synthase) and anti-Alzheimer's agents such as donepezil, tacrine, COX-2 inhibitors, propentofylline or metryfonate.

The compounds of formula (I) of the present invention may be administered in combination with an osteoporosis drug such as raloxifene, droloxifene, lasofoxifene or Fosomax and an immunosuppressant such as FK-506, rapamycin, cyclosporine, azathioprine and methotrexate.

Examples

Example 1

General synthetic method I

General procedure for preparation of XInt 01:

to indole derivative X (1eq) in TEA and DMAP in DCM at room temperature was added (Boc)₂O and the resulting reaction mixture was stirred at room temperature. After 1 hour, the reaction mixture was diluted with water and extracted 3 times with DCM. The combined DCM layers were washed with 1N HCl solution and Na₂SO₄Dried and concentrated under reduced pressure to give the desired XInt01 as a liquid.

General procedure for preparation of XInt 02:

ac to stirred XINT01₂O solution at-78 deg.C for 15 minAdding ice-cold fuming HNO into the bell₃Ac of (a)₂And (4) O solution. The reaction mixture was slowly warmed to room temperature and stirring was continued. After 16 h, it was diluted with ice water and extracted 3 times with EtOAc. The combined EtOAc layers were washed with brine, Na₂SO₄Dried and concentrated under reduced pressure. Purification by flash chromatography (SiO)₂100-mesh, 1% EtOAc in petroleum ether) to give the desired XInt02 as a liquid.

General procedure for preparation of XInt 03:

to a stirred solution of XINT02 in DCM at 0 deg.C was added TFA and the resulting reaction mixture was slowly warmed to elevated temperature and stirred. After 2 hours, the reaction mixture was concentrated under reduced pressure to give XINT 03.

General procedure for preparation of XInt 04:

to a stirred dry DMF solution of XINT03 was added 2-chloroethylbenzoate (Y05-OBz) and K₂CO₃And the resulting reaction mixture was heated to 60 ℃. After 16 h, the reaction mixture was diluted with ice water and extracted 3 times with DCM. The combined DCM layers were washed with Na₂SO₄Dried and concentrated under reduced pressure. Purification by trituration afforded XInt04 as a solid.

General procedure for preparation of XInt 05:

to a stirred solution of XInt04 in MeOH was added indium, (Boc)₂O and NH₄And (4) Cl. The reaction mixture was heated to reflux. After 30 minutes it was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was diluted with water and extracted 3 times with EtOAc. The combined EtOAc layers were washed with brine, Na₂SO₄Dried and concentrated under reduced pressure. Purification by flash chromatography (SiO)₂100-mesh 200, 5% EtOAc in petroleum ether) to give XInt05 as a solid.

General procedure for preparation of XInt 06:

to a stirred solution of XINT05 in DCM at 0 deg.C was added TFA and the resulting reaction mixture was slowly warmed to room temperature. After 2 hours the reaction mixture was concentrated under reduced pressure to give XINT 06.

General procedure for making XZInt 07:

to a stirred solution of XINT06 in THF (3.0mL) at 0 deg.C was added TFA (296mg, 2.93mmol) and acid chloride Z (e.g., cyclohexylacetic acid, cycloheptylacetic acid, cyclohexylpropionic acid, or cycloheptylpropionic acid) and the resulting reaction mixture was slowly warmed to room temperature. After 30 min the reaction mixture was diluted with water and extracted 3 times with EtOAc. The combined EtOAc layers were washed with brine, Na₂SO₄Dried and concentrated under reduced pressure. Purification by trituration afforded XZInt07 as a solid.

General procedure for the preparation of XZ:

to a stirred solution of XZInt07 in MeOH was added K₂CO₃And the resulting reaction mixture was stirred at room temperature. After 30 minutes, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (SiO)₂100-200 mesh, 50% EtOAc in petroleum ether) to give XZ as a solid.

Example 2

N- (4-chloro-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 1, wherein X is 4-chloroindole and Z is cycloheptylacetyl chloride. The molecular formula is as follows: c₁₉H₂₅ClN₂O₂(ii) a Molecular weight: 348.9, respectively; mass/charge ratio: 348.2 (100.0%), 350.2 (34.6%), 349.2 (21.7%), 351.2 (7.2%); elemental analysis: c, 65.41; h, 7.22; cl, 10.16; n, 8.03; and O, 9.17.

Example 3

N- (4-bromo-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 1, wherein X is 4-bromoindole and Z is cycloheptylacetyl chloride. The molecular formula is as follows: c₁₉H₂₅BrN₂O₂(ii) a Molecular weight: 393.3 of the total weight of the mixture; mass/charge ratio: 392.1 (100.0%), 394.1 (99.9%), 393.1 (21.7%), 395.1 (21.3%), 396.1 (2.6%); elemental analysis: c, 58.02; h, 6.41; br, 20.32; n, 7.12; and O, 8.14.

Example 4

N- (4-chloro-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 1, wherein X is 4-chloroindole and Z is cyclohexylacetyl chloride. The molecular formula is as follows: c₁₈H₂₃ClN₂O₂(ii) a Molecular weight: 334.8 parts of a main body; mass/charge ratio: 334.1 (100.0%), 336.1 (32.5%), 335.1 (20.3%), 337.1 (6.6%), 336,2 (1.9%); elemental analysis: c, 64.57; h, 6.92; cl, 10.59; n, 8.37; and O, 9.56.

Example 5

N- (4-bromo-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 1, whichWherein X is 4-bromoindole and Z is cyclohexylacetyl chloride. The molecular formula is as follows: c₁₈H₂₃BrN₂O₂(ii) a Molecular weight: 379.3, respectively; mass/charge ratio: 378.1 (100.0%), 380.1 (99.7%), 379.1 (20.5%), 381.1 (20.2%), 382.1 (2.4%); elemental analysis: c, 57.00; h, 6.11; br, 21.07; n, 7.39; and O, 8.44.

Example 6

N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloroindole and Z is cycloheptylacetyl chloride. The molecular formula is as follows: c₂₀H₂₇ClN₂O₂(ii) a Molecular weight: 362.9, respectively; mass/charge ratio: 362.2 (100.0%), 364.2 (34.8%), 363.2 (22.8%), 365.2 (7.5%); elemental analysis: c, 66.19; h, 7.50; cl, 9.77; n, 7.72; and O, 8.82.

Example 7

N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromoindole and Z is cycloheptylacetyl chloride. The molecular formula is as follows: c₂₀H₂₇BrN₂O₂(ii) a Molecular weight: 407.3, respectively; mass/charge ratio: 408.1 (100.0%), 406.1(99, 8%), 407.1(22, 7%), 409.1 (22.4%), 410.1 (2.8%); elemental analysis: c, 58.97; h, 6.68; br, 19.62; n, 6.88; and O, 7.86.

Example 8

N- (4-chloro-1- (2-hydroxyethyl) -1H-indol-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 1, wherein X is 4-chloroindole and Z is cyclohexylpropionyl chloride. The molecular formula is as follows: c₁₉H₂₅ClN₂O₂(ii) a Molecular weight: 348.9, respectively; mass/charge ratio: 348.2 (100.0%), 350.2 (34.6%), 349.2 (21.7%), 351.2 (7.2%); elemental analysis: c, 65.41; h, 7.22; cl, 10.16; n, 8.03; and O, 9.17.

Example 9

N- (4-bromo-1- (2-hydroxyethyl) -1H-indol-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 1, wherein X is 4-bromoindole and Z is cyclohexylpropionyl chloride. The molecular formula is as follows: c₁₉H₂₅BrN₂O₂(ii) a Molecular weight: 393.3 of the total weight of the mixture; mass/charge ratio: 392.1 (100.0%), 394.1 (99.9%), 393.1 (21.7%), 395.1 (21.3%), 396.1(2, 6%); elemental analysis: c, 58.02; h, 6.41; br, 20.32; n, 7.12; and O, 8.14.

Example 10

N- (4-chloro-1- (2-hydroxyethyl) -1H-indol-3-yl) -3-cycloheptylpropanamide

Synthesized according to the method disclosed in example 1, wherein X is 4-chloroindole and Z is cycloheptylpropionyl chloride. Is divided intoThe subformula: c₂₀H₂₇ClN₂O₂(ii) a Molecular weight: 362.9, respectively; mass/charge ratio: 362.2 (100.0%), 364.2 (34.8%), 363.2 (22.8%), 365.2 (7.5%); elemental analysis: c, 66.19; h, 7.50; cl, 9.77; n, 7.72; and O, 8.82.

Example 11

N- (4-bromo-1- (2-hydroxyethyl) -1H-indol-3-yl) -3-cycloheptylpropanamide

Synthesized according to the method disclosed in example 1, wherein X is 4-bromoindole and Z is cycloheptylpropionyl chloride. The molecular formula is as follows: c₂₀H₂₇BrN₂O₂(ii) a Molecular weight: 407.3, respectively; mass/charge ratio: 408.1 (100.0%), 406.1 (99.8%), 407.1 (22.7%), 409.1 (22.4%), 410.1 (2.8%); elemental analysis: c, 58.97; h, 6.68; br, 19.62; n, 6.88; and O, 7.86.

Example 12

N- (4-chloro-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 1, wherein X is 4-chloro-1H-pyrrolo [2, 3-b ]]Pyridine, Z is cyclohexylacetyl chloride. The molecular formula is as follows: c₁₇H₂₂ClN₃O₂(ii) a Molecular weight: 335.8; mass/charge ratio: 335.1 (100.0%), 337.1 (34.2%), 336.1 (19.8%), 338.1 (6.4%); elemental analysis: c, 60.80; h, 6.60; cl, 10.56; n, 12.51; and O, 9.53.

Example 13

N- (4-bromo-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 1, wherein X is 4-bromo-1H-pyrrolo [2, 3-b ]]Pyridine, Z is cyclohexylacetyl chloride. The molecular formula is as follows: c₁₇H₂₂BrN₃O₂(ii) a Molecular weight: 380.3, respectively; mass/charge ratio: 379.1 (100.0%), 381.1 (99.6%), 380.1 (19.8%), 382.1 (19.5%), 383.1 (2.2%); elemental analysis: c, 53.69; h, 5.83; br, 21.01; n, 11.05; o, 8.41.

Example 14

N- (4-chloro-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 1, wherein X is 4-chloro-1H-pyrrolo [2, 3-b ]]Pyridine, Z is cycloheptylacetyl chloride. The molecular formula is as follows: c₁₈H₂₄ClN₃O₂(ii) a Molecular weight: 349.9; mass/charge ratio 349.2 (100.0%), 351.2 (34.5%), 350.2 (20.9%), 352.2 (6.6%); elemental analysis: c, 61.79; h, 6.91; cl, 10.13; n, 12.01; and O, 9.15.

Example 15

N- (4-bromo-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide

As disclosed in example 1The synthesis of the compound is carried out by the open method, wherein X is 4-bromo-1H-pyrrolo [2, 3-b ]]Pyridine, Z is cycloheptylacetyl chloride. The molecular formula is as follows: c₁₈H₂₄BrN₃O₂(ii) a Molecular weight: 394.3, respectively; mass/charge ratio: 393.1 (100.0%), 395.1 (99.8%), 394.1 (20.9%), 396.1 (20.6%), 397.1 (2.4%); elemental analysis: c, 54.83; h, 6.13; br, 20.26; n, 10.66; and O, 8.12.

Example 16

N- (4-chloro-1- (2-hydroxypropyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloro-1H-pyrrolo [2, 3-b ]]Pyridine, Z is cycloheptylacetyl chloride. The molecular formula is as follows: c₁₉H₂₆ClN₃O₂(ii) a Molecular weight: 363.9; mass/charge ratio: 363.2 (100.0%), 365.2 (34.7%), 364.2 (22.0%), 366.2 (7.3%); elemental analysis: c, 62.71; h, 7.20; cl, 9.74; n, 11.55; o, 8.79.

Example 17

N- (4-bromo-1- (2-hydroxypropyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromo-1H-pyrrolo [2, 3-b ]]Pyridine, Z is cycloheptylacetyl chloride. The molecular formula is as follows: c₁₉H₂₆BrN₃O₂(ii) a Molecular weight: 408.3, respectively; mass/charge ratio: 409.1 (100.0%), 407.1 (100.0%), 408.1 (22.0%), 410,1 (21.7%), 411.1 (2.7%); elemental analysis: c, 55.89; h, 6.42; br, 19.57; n, 10.29；O，7.84。

Example 18

N- (4-chloro-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 1, wherein X is 4-chloro-1H-pyrrolo [2, 3-b ]]Pyridine, Z is cyclohexylpropionyl chloride. The molecular formula is as follows: c₁₈H₂₄ClN₃O₂(ii) a Molecular weight: 349, 9; mass/charge ratio: 349.2 (100.0%), 351.2 (34.5%), 350.2 (20.9%), 352.2 (6.6%); elemental analysis: c, 61.79; h, 6.91; cl, 10.13; n, 12.01; and O, 9.15.

Example 19N- (4-bromo-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 1, wherein X is 4-bromo-1H-pyrrolo [2, 3-b ]]Pyridine, Z is cyclohexylpropionyl chloride. The molecular formula is as follows: c₁₈H₂₄BrN₃O₂(ii) a Molecular weight: 394.3, respectively; mass/charge ratio: 393.1 (100.0%), 395.1 (99.8%), 394.1 (20.9%), 396.1 (20.6%), 397.1 (2.4%); elemental analysis: c, 54.83; h, 6.13; br, 20.26; n, 10.66; and O, 8.12.

Further preferred examples include the following compounds:

example 20

General Process for preparing XZInt01

To a solution of indole derivative X (6.6mmol) in dry benzene (10mL) at 0 deg.C was added a solution of acid chloride Z (e.g. 2-cyclohexylacetyl chloride, 2-cycloheptylacetyl chloride, 2-cyclohexylpropionyl chloride, 2-cycloheptylpropionyl chloride) (13mmol) in dry benzene (10 mL). Dropwise adding SnCl at 0 DEG C₄(26.49mM) in dry benzene (15 mL). The reaction mixture was warmed to room temperature and held for 3 hours. The mixture was poured into 5% HCl solution (50mL) and ethyl acetate (100mL) and stirred for 10 min. The organic layer was separated and washed with water (50mL), saturated NaHCO₃The solution (50mL), brine (50mL) was washed, dried over anhydrous sodium sulfate and concentrated. The crude product was purified by column chromatography on silica eluting with ethyl acetate and chloroform to give pure xzip 01.

General Process for preparing XZInt02

To a stirred solution of XZInt01(2.25mmol) in methanol (10ml) was added NH at room temperature₂OH HCl (4.49mM) and pyridine (6.74 mM). The mixture was then refluxed for 2 hours. The methanol was distilled off and the resulting residue was dissolved in ethyl acetate (75mL), washed with water (50mL), brine (50mL), dried over anhydrous sodium sulfate and concentrated. The crude product was purified by column chromatography on silica eluting with ethyl acetate and hexanes to provide XZInt 02.

General Process for preparing XZInt03

A solution of XZInt02(1.8mmol) in TFA (15mL) was refluxed for 5 hours. The reaction mixture was then concentrated to give a residue. The residue was dissolved in ethyl acetate (100mL) and washed with water (50mL), saturated sodium bicarbonate solution (50mL), water (50mL) and brine (50 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column eluting with ethyl acetate and hexanes to provide XZInt 03.

General Process for preparing XZ

XZInt03(0.33mmol), (R) -2-epoxypropane or alkaneIodonium (3.3mmol) and Cs₂CO₃A mixture of (1.64mmol) in dry DMF (2mL) was maintained in a microwave at 120 ℃ for 20 minutes. The reaction mixture was then poured into water and extracted with ethyl acetate (100 mL). The organic layer was washed with water (50mL), brine (50mL), dried over anhydrous sodium sulfate and concentrated. The crude product was purified through a silica gel column to give XZ.

Example 21

General synthetic method III

General Process for preparing XZInt01

To a solution of azaindole derivative X (6.5mmol) in dry benzene was added acid chloride Z (e.g. 2-cyclohexylacetyl chloride, 2-cycloheptylacetyl chloride, 2-cyclohexylpropionyl chloride, 2-cycloheptylpropionyl chloride) (13mmol) at 0 ℃ and stirred for 10 min. SnCl was added dropwise thereto at 0 deg.C₄(26.2 mmol). The reaction mixture was slowly warmed to room temperature and stirred for 3 hours. The reaction mixture was poured into 2N HCl (50mL) and extracted with ethyl acetate (100 mL). The organic layer was washed with water (50mL), saturated NaHCO₃The solution (50m), brine (50mL) was washed and dried over anhydrous sodium sulfate to give XZInt 01.

General Process for preparing XZInt02

To a stirred solution of XZInt01(2.8mmol) in methanol (16mL) at room temperature was added NH₂OH HCl (5.75mmol) and pyridine (8.6 mmol). The reaction mixture was heated to reflux for 2 h, evaporated, the residue diluted with water (75mL) and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with brine (50mL), dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column eluting with ethyl acetate and hexanes to provide XZInt 02.

General Process for preparing XZInt03

A solution of XZInt02(1.7mmol) in TFA (15ml) was refluxed for 5 hours. The reaction mixture was concentrated, the residue diluted with water (50mL) and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with saturated NaHCO₃The solution (50mL), brine (50mL), was washed, dried over anhydrous sodium sulfate and concentrated, and the crude product was purified through a silica gel column eluting with ethyl acetate and hexanes to provide XZInt 03.

General Process for preparing XZInt04

To a stirred solution of XZInt03(1.2mmol) in DMF at room temperature was added Cs₂CO₃(3.6mmol) and stirred for 15 min. Chloroethyl benzoate (1.8mmol) was added and the mixture was heated to 80 ℃ and stirred for 16 h. The reaction mixture was poured into ice water and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with brine (50mL), dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column eluting with ethyl acetate and hexanes to provide XZInt 04.

General Process for preparing XZ

To a solution of XZInt04(0.3mmol) in methanol (10mL) at room temperature was added K₂CO₃(1.0mmol) and stirred for 1 hour. The reaction mixture was concentrated, the residue diluted with water (10mL) and extracted with ethyl acetate (2 × 25 mL). The ethyl acetate layer was washed with brine (20mL), dried over anhydrous sodium sulfate and concentrated. Trituration of the crude product with diethyl ether yielded XZ.

Example 22

General synthetic Process VI

Preparation of diethyl 2- (4-chloro-3 (-2-cycloheptylacetamino) -indol-1-yl) malonate (step 4)

To a solution of sodium azide (4.2g, 64.6mmol) in 70% aqueous ethanol (150mL) was added p-toluenesulfonyl chloride (11.8g, 65.5mmol) at room temperature and stirred at room temperature for 1 hour. Diethyl malonate (10g, 62.5mm) and triethylamine (6.3g, 65.5mm) were added and the reaction mixture was stirred at room temperature for 1.5 hours. The reaction mixture was poured into water and extracted with hexane (2 × 50 mL). The combined organic layers were washed once with brine solution (50mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give diethyl 2- (4-chloro-3 (-2-cycloheptylacetylamino) -indol-1-yl) -malonate (9g, 77.5%) as a yellow oil.

General Process for preparing XZInt04

To a solution of XZInt03(1.6 mmol; see example 20) in DCM (5ml) was added rhodium acetate (0.32mmol) and diethyl 2- (4-chloro-3 (-2-cycloheptylacetylamino) -indol-1-yl) -malonate (4.0mmol) and the reaction mixture was stirred at room temperature for 12 h. Water (50ml) was added, followed by extraction with ethyl acetate (50mlx 3). The combined organic layers were washed once with brine solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash column chromatography (using 100-200 mesh silica gel) and the product XZInt04 was eluted with 7% ethyl acetate/chloroform.

General Process for preparing XZ

To a stirred solution of XZInt04(0.54mmol) in methanol under nitrogen at 0 ℃ was added sodium borohydride (2.6mmol) and the reaction mixture was warmed to room temperature and then stirred for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in water (25mL) and extracted with ethyl acetate (2 × 25 mL). The combined organic layers were washed with brine (20mL) and dried over anhydrous sodium sulfate and concentrated. Purification by flash column chromatography eluting with 100-200 mesh silica gel (3% methanol/chloroform) afforded XZ.

Example 23

General synthetic method V

General Process for preparing XZInt04

To a solution of XZInt03(1.9 mmol; see example 20) in DMF (6ml) at room temperature was added K₂CO₃(5.9mmol) and the reaction mixture was stirred for 10 minutes. R- (-) epichlorohydrin (5.9mmol) was added under a nitrogen atmosphere and the reaction mixture was heated at 85 ℃ for 12 hours. The reaction mixture was then poured into ice water and extracted with ethyl acetate (50mlx 3). The combined organic layers were washed once with brine solution (25ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified through a silica gel column (100-200 mesh) eluting with ethyl acetate/hexanes to afford XZInt 04.

General Process for preparing XZ

To a solution of XZInt04(0.55mmol) in ethanol (5ml) was added aqueous ammonia or an alkylamine (10ml) and the reaction mixture was refluxed for 1 hour. The reaction mixture was concentrated under reduced pressure and co-distilled with ethanol, then triturated with pentane and diethyl ether to give the crude product as a solid after purification by HPLC. Treatment with dioxane-HCl gave the HCl salt of XZ.

Example 24

N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloroindole and Z is 2-cyclohexylacetyl chloride. The molecular formula is as follows: c₁₉H₂₅ClN₂O₂(ii) a Molecular weight: 348.9, respectively; mass/charge ratio: 348.2 (100.0%), 350.2 (34.6%), 349.2 (21.7%), 351.2 (7.2%); elemental analysis: c, 65.41; h, 7.22; cl, 10.16; n, 8.03; and O, 9.17.

Example 25

N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromoindole and Z is 2-cyclohexylacetyl chloride. The molecular formula is as follows: c₁₉H₂₅BrN₂O₂(ii) a Molecular weight: 393.3 of the total weight of the mixture; mass/charge ratio: 392.1 (100.0%), 394.1 (99.9%), 393.1 (21.7%), 395.1 (21.3%), 396.1 (2.6%); elemental analysis: c, 58.02; h, 6.41; br, 20.32; n, 7.12; and O, 8.14.

Example 26

2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) acetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloroindole and Z is bicyclo [2.2.2]Octane-1-yl acetyl chloride. The molecular formula is as follows: c₂₁H₂₇ClN₂O₂(ii) a Molecular weight: 374.9, respectively; mass/charge ratio: 374.2 (100.0%), 376.2 (35.1%), 375.2 (23.8%), 377.2 (7.9%), 378.2 (1.0%); elemental analysis: c, 67.28; h, 7.26; cl, 9.46; n, 7.47; and O, 8.54.

Example 27

2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) acetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromoindole and Z is bicyclo [2.2.2]Octane-1-yl acetyl chloride. The molecular formula is as follows: c₂₁H₂₇BrN₂O₂(ii) a Molecular weight: 419.4, respectively; mass/charge ratio: 420.1 (100.0%), 418.1 (99.6%), 419.1 (23.7%), 421.1 (23.4%), 422.1 (3.1%); elemental analysis: c, 60.15; h, 6.49; br, 19.05; n, 6.68; and O, 7.63.

Example 28

N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloroindole and Z is 2-cyclohexylpropionyl chloride. The molecular formula is as follows: c₂₀H₂₇ClN₂O₂(ii) a Molecular weight: 362.9, respectively; mass/charge ratio: 362.2 (100.0%), 364.2 (34.8%), 363.2 (22.8%), 365.2 (7.5%); elemental analysis: c, 66.19; h, 7.50; cl, 9.77; n, 7.72; and O, 8.82.

Example 29

N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromoindole and Z is 2-cyclohexylpropionyl chloride. The molecular formula is as follows: c₂₀H₂₇BrN₂O₂(ii) a Molecular weight: 407.3, respectively; mass/charge ratio: 408.1 (100.0%), 406.1 (99.8%), 407.1 (22.7%), 409.1 (22.4%), 410.1 (2.8%); elemental analysis: c, 58.97; h, 6.68; br, 19.62; n, 6.88; and O, 7.86.

Example 30

N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cycloheptylpropanamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloroindole and Z is 2-cycloheptylpropionyl chloride. The molecular formula is as follows: c₂₁H₂₉ClN₂O₂(ii) a Molecular weight: 376.9; mass/charge ratio: 376.2 (100.0%), 378.2 (35.1%), 377.2 (23.9%), 379.2 (7.9%), 380.2 (1.0%); elemental analysis: c, 66.92; h, 7.76; cl, 9.41; n, 7.43; o, 8.49.

Example 31

N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cycloheptylpropanamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromoindole and Z is 2-cycloheptylpropionyl chloride. The molecular formula is as follows: c₂₁H₂₉BrN₂O₂(ii) a Molecular weight: 421.4; mass/charge ratio: 422.1 (100.0%), 420.1 (99.7%), 421.1 (23.8%), 423.1 (23.2%), 424.1 (3.0%); elemental analysis: c, 59.86; h, 6.94; br, 18.96; n, 6.65; and O, 7.59.

Example 32

N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 22, wherein X is 4-chloroindole and Z is 2-cyclohexylacetyl chloride. The molecular formula is as follows: c₁₉H₂₅ClN₂O₃(ii) a Molecular weight: 364.9; mass/charge ratio: 364.2 (100.0%), 366.2 (34.8%), 365.2 (21.7%), 367.2 (7.0%); elemental analysis: c, 62.54; h, 6.91; cl, 9.72; n, 7.68; and O, 13.16.

Example 33

N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 22, wherein X is 4-bromoindole and Z is 2-cyclohexylacetyl chloride. The molecular formula is as follows: c₁₉H₂₅BrN₂O₃(ii) a Molecular weight: 409.3, respectively; mass/charge ratio: 410.1 (100.0%), 408.1 (99.9%), 409.1 (21.7%), 411.1 (21.4%), 412.1 (2.8%); elemental analysis C, 55.75; h, 6.16; br, 19.52; n, 6.84; o, 11.73.

Example 34

N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 22, wherein X is 4-chloroindole and Z is 2-cycloheptylacetyl chloride. The molecular formula is as follows: c₂₀H₂₇ClN₂O₃(ii) a Molecular weight: 378.9, respectively; mass/charge ratio 378.2 (100.0%), 380.2 (35.1%), 379.2 (22.8%), 381.2 (7.6%), 382.2 (1.0%); elemental analysis: c, 63.40; h, 7.18; cl, 9.36; n, 7.39; o, 12.67.

Example 35

N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 22, wherein X is 4-bromoindole and Z is 2-cycloheptylacetyl chloride. The molecular formula is as follows: c₂₀H₂₇BrN₂O₃(ii) a Molecular weight 423.3; mass/charge ratio: 424.1 (100.0%), 422.1 (99.6%), 423.1 (22.7%), 425.1 (22.4%), 426.1 (3.0%); elemental analysis: c, 56.74; h, 6.43; br, 18.87; n, 6.62; o, 11.34.

Example 36

2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) acetamide

Synthesized according to the method disclosed in example 22, wherein X is 4-chloroindole and Z is bicyclo [2.2.2]Octane-1-yl acetyl chloride. The molecular formula is as follows: c₂₁H₂₇ClN₂O₃(ii) a Molecular weight: 390.9, respectively; mass/charge ratio: 390.2 (100.0%), 392.2 (35.3%), 391.2 (23.9%), 393.2 (8.0%), 394.2 (1.1%); elemental analysis: c, 64.52; h, 6.96; cl, 9.07; n, 7.17; o, 12.28.

Example 37

2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) acetamide

Synthesized according to the method disclosed in example 22, wherein X is 4-bromoindole and Z is bicyclo [2.2.2]Octane-1-yl acetyl chloride. The molecular formula is as follows: c₂₁H₂₇BrN₂O₃(ii) a Molecular weight: 435.4; mass/charge ratio: 436.1 (100.0%), 434.1 (99.4%), 435.1 (23.7%), 437.1 (23.4%), 438.1 (3.3%); elemental analysis: c, 57.94; h, 6.25; br, 18.35; n, 6.43; and O, 11.03.

Example 38

N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 22, wherein X is 4-chloroindole and Z is 2-cyclohexylpropionyl chloride. The molecular formula is as follows: c₂₀H₂₇ClN₂O₃(ii) a Molecular weight: 378.9, respectively; mass/charge ratio: 378.2 (100.0%), 380.2 (35.1%), 379.2 (22.8%), 381.2 (7.6%), 382.2 (1.0%); elemental analysis: c, 63.40; h, 7.18; cl, 9.36; n, 7.39; o, 12.67.

Example 39

N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 22, wherein X is 4-bromoindole and Z is 2-cyclohexylpropionyl chloride. The molecular formula is as follows: c₂₀H₂₇BrN₂O₃(ii) a Molecular weight: 423.3, respectively; mass/charge ratio: 424.1 (100.0%), 422.1 (99.6%) 423.1 (22.7%), 425.1 (22.4%), 426.1 (3.0%); elemental analysis: c, 56.74; h, 6.43; br, 18.87; n, 6.62; o, 11.34.

Example 40

N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cycloheptylpropanamide

Synthesized according to the method disclosed in example 22, wherein X is 4-chloroindole and Z is 2-cycloheptylpropionyl chloride. The molecular formula is as follows: c₂₁H₂₉ClN₂O₃(ii) a Molecular weight: 392.9, respectively; mass/charge ratio: 392.2 (100.0%), 394.2 (35.3%), 393.2 (23.9%), 395.2 (8.0%), 396.2(1, 1%); elemental analysis: c, 64.19; h, 7.44; cl, 9.02; n, 7.13; o, 12.22.

EXAMPLE 41

N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cycloheptylpropanamide

Synthesized according to the method disclosed in example 22, wherein X is 4-bromoindole and Z is 2-cycloheptylpropionyl chloride. The molecular formula is as follows: c₂₁H₂₉BrN₂O₃(ii) a Molecular weight: 437.4, respectively; mass/charge ratio: 438.1 (100.0%), 436.1 (99.4%), 437.1 (23.8%), 439.1 (23.4%), 440.1 (3.3%); elemental analysis: c, 57.67; h, 6.68; br, 18.27; n, 6.40; o, 10.97.

Example 42

N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-chloroindole and Z is 2-cyclohexylacetyl chloride. The molecular formula is as follows: c₁₉H₂₆ClN₃O₂(ii) a Molecular weight: 363.9; mass/charge ratio: 363.2 (100.0%), 365.2 (34.7%), 364.2 (22.0%), 366.2 (7.3%); elemental analysis: c, 62.71; h, 7.20; cl, 9.74; n, 11.55; o, 8.79.

Example 43

N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-bromoindole and Z is 2-cyclohexylacetyl chloride. The molecular formula is as follows: c₁₉H₂₆BrN₃O₂(ii) a Molecular weight: 408.3, respectively; mass/charge ratio: 409.1 (100.0%), 407.1 (100.0%), 408.1 (22.0%), 410.1 (21.7%), 411.1 (2.7%); elemental analysis: c, 55.89; h, 6.42; br, 19.57; n, 10.29; and O, 7.84.

Example 44

N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-chloroindole and Z is 2-cycloheptylacetyl chloride. The molecular formula is as follows: c₂₀H₂₈ClN₃O₂(ii) a Molecular weight: 377.9, respectively; quality ofCharge ratio: 377.2 (100.0%), 379.2 (34.9%), 378.2 (23.1%), 380.2 (7.7%); elemental analysis: c, 63.56; h, 7.47; cl, 9.38; n, 11.12; and O, 8.47.

Example 45

N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-bromoindole and Z is 2-cycloheptylacetyl chloride. The molecular formula is as follows: c₂₀H₂₈BrN₃O₂(ii) a Molecular weight: 422.4, respectively; mass/charge ratio: 423.1 (100.0%), 421.1 (99.8%), 422.1 (23.1%), 424.1 (22.7%), 425.1 (2.9%); elemental analysis: c, 56.87; h, 6.68; br, 18.92; n, 9.95; and O, 7.58.

Example 46

N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -2- (bicyclo [2.2.2] octan-1-yl) acetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-chloroindole and Z is bicyclo [2.2.2]Octane-1-yl acetyl chloride. The molecular formula is as follows: c₂₁H₂₈ClN₃O₂(ii) a Molecular weight: 389.9; mass/charge ratio: 389.2 (100.0%), 391.2 (35.2%), 390.2 (24.2%), 392.2 (8.0%), 393.2 (1.1%); elemental analysis: c, 64.69; h, 7.24; cl, 9.09; n, 10.78; o, 8.21.

Example 47

N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -2- (bicyclo [2.2.2] octan-1-yl) acetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-bromoindole and Z is bicyclo [2.2.2]Octane-1-yl acetyl chloride. The molecular formula is as follows: c₂₁H₂₈BrN₃O₂(ii) a Molecular weight: 434.4; mass/charge ratio: 435.1 (100.0%), 433.1 (99.5%), 434.1 (24.1%), 436.1 (23.7%), 437.1 (3.1%); elemental analysis: c, 58.07; h, 6.50; br, 18.40; n, 9.67; o, 7.37.

Example 48

N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 23, wherein X is 4-chloroindole and Z is 2-cyclohexylpropionyl chloride. The molecular formula is as follows: c₂₀H₂₈ClN₃O₂(ii) a Molecular weight: 377.9, respectively; mass/charge ratio: 377.2 (100.0%), 379.2 (34.9%), 378.2 (23.1%), 380.2 (7.7%); elemental analysis: c, 63.56; h, 7.47; cl, 9.38; n, 11.12; and O, 8.47.

Example 49

N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 23, wherein X is 4-bromoindole and Z is 2-cyclohexylpropionyl chloride. The molecular formula is as follows: c₂₀H₂₈BrN₃O₂(ii) a Molecular weight: 422.4, respectively; mass/charge ratio: 423.1 (100.0%), 421.1 (99.8%), 422.1 (23.1%), 424.1 (22.7%), 425.1 (2.9%); elemental analysis: c, 56.87; h, 6.68; br, 18.92; n, 9.95; and O, 7.58.

Example 50

N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -3-cycloheptylpropanamide

Synthesized according to the method disclosed in example 23, wherein X is 4-chloroindole and Z is 2-cycloheptylpropionyl chloride. The molecular formula is as follows: c₂₁H₃₀ClN₃O₂(ii) a Molecular weight: 391.9, respectively; mass/charge ratio: 391.2 (100.0%), 393.2 (35.2%), 392.2 (24.2%), 394.2 (8.1%), 395.2 (1.1%); elemental analysis: c, 64.35; h, 7.72; cl, 9.05; n, 10.72; and O, 8.16.

Example 51

N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -3-cycloheptylpropanamide

Synthesized according to the method disclosed in example 23, wherein X is 4-bromoindole and Z is 2-cycloheptylpropionyl chloride. The molecular formula is as follows: c₂₁H₃₀BrN₃O₂(ii) a Molecular weight: 436.4; mass/charge ratio: 437.2 (100.0%), 435.2 (99.5%), 436.2 (23.0%), 438.2 (22.7%), 439.2 (3.1%), 436.1 (1.1%), 438.1 (1.1%); elemental analysis: c, 57.80; h, 6.93; br, 18.31; n, 9.63; o, 7.33.

Example 52

N- (4-chloro-1-methyl-1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloroindole and Z is 2-cyclohexylacetyl chloride. The molecular formula is as follows: c₁₇H₂₁ClN₂O; molecular weight: 304.8; mass/charge ratio: 304.1 (100.0%), 306.1 (33.9%), 305.1 (19.4%), 307.1 (6.3%); elemental analysis: c, 66.99; h, 6.94; cl, 11.63; n, 9.19; and O, 5.25.

Example 53

N- (4-bromo-1-methyl-1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromoindole and Z is 2-cyclohexylacetyl chloride. The molecular formula is as follows: c₁₇H₂₁BrN₂O; molecular weight: 349.3; mass/charge ratio: 348.1 (100.0%), 350.1 (99.3%), 349.1 (19.4%), 351.1 (19.0%), 352.1 (1.9%); elemental analysis: c, 58.46; h, 6.06; br, 22.88; n, 8.02; and O, 4.58.

Example 54

N- (4-chloro-1-methyl-1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloroindole and Z is 2-cycloheptylacetyl chloride. The molecular formula is as follows: c₁₈H₂₃ClN₂O; molecular weight: 318.8, respectively; mass/charge ratio: 318.1 (100.0%), 320.1 (32.0%), 319.2 (19.8%), 321.2 (6.5%), 320.2 (2.2%); elemental analysis: c, 67.81; h, 7.27; cl, 11.12; n, 8.79; and O, 5.02.

Example 55

N- (4-bromo-1-methyl-1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromoindole and Z is 2-cycloheptylacetyl chloride. The molecular formula is as follows: c₁₈H₂₃BrN₂O; molecular weight: 363.3; mass/charge ratio: 362.1 (100.0%), 364.1 (99.5%), 363.1 (20.5%), 365.1 (20.1%), 366.1 (2.2%); elemental analysis: c, 59.51; h, 6.38; br, 21.99; n, 7.71; and O, 4.40.

Example 56

2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1-methyl-1H-indol-3-yl) acetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloroindole and Z is bicyclo [2.2.2]Octane-1-yl acetyl chloride. The molecular formula is as follows: c₁₉H₂₃ClN₂O; molecular weight: 330.9, respectively; mass/charge ratio: 330.1 (100.0%), 332.1 (32.0%), 331.2 (20.9%), 333.2 (6.9%), 332.2 (2.4%); elemental analysis: c, 68.97; h, 7.01; cl, 10.72; n, 8.47; and O, 4.84.

Example 57

2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1-methyl-1H-indol-3-yl) acetamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromoindole and Z is bicyclo [2.2.2]Octane-1-yl acetyl chloride. The molecular formula is as follows: c₁₉H₂₃BrN₂O; molecular weight: 375.3 of the total weight of the product; mass/charge ratio: 374.1 (100.0%), 376.1 (99.7%), 375.1 (21.6%), 377.1 (21.2%), 378.1 (2.4%); elemental analysis: c, 60.81; h, 6.18; br, 21.29; n, 7.46; and O, 4.26.

Example 58

N- (4-chloro-1-methyl-1H-indol-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloroindole and Z is 2-cyclohexylpropionyl chloride. The molecular formula is as follows: c₁₈H₂₃ClN₂O; molecular weight: 318.8, respectively; mass/charge ratio: 318.1 (100.0%), 320.1 (32.0%), 319.2 (19.8%), 321.2 (6.5%), 320.2 (2.2%); elemental analysis: c, 67.81; h, 7.27; cl, 11.12; n, 8.79; and O, 5.02.

Example 59

N- (4-bromo-1-methyl-1H-indol-3-yl) -3-cyclohexylpropionamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromoindole and Z is 2-cyclohexylpropionyl chloride. The molecular formula is as follows: c₁₈H₂₃BrN₂O; molecular weight: 363.3; mass/charge ratio: 362.1 (100.0%), 364.1 (99.5%), 363.1 (20.5%), 365.1 (20.1%), 366.1 (2.2%); elemental analysis: c, 59.51; h, 6.38; br, 21.99; n, 7.71; and O, 4.40.

Example 60

N- (4-chloro-1-methyl-1H-indol-3-yl) -3-cycloheptylpropanamide

Synthesized according to the method disclosed in example 20, wherein X is 4-chloroindole and Z is 2-cycloheptylpropionyl chloride. The molecular formula is as follows: c₁₉H₂₅ClN₂O; molecular weight: 332.9; mass/charge ratio: 332.2 (100.0%), 334.2 (34.4%), 333.2 (21.6%), 335.2 (7.1%); elemental analysis: c, 68.56; h, 7.57; cl, 10.65; n, 8.42; and O, 4.81.

Example 61

N- (4-bromo-1-methyl-1H-indol-3-yl) -3-cycloheptylpropanamide

Synthesized according to the method disclosed in example 20, wherein X is 4-bromoindole and Z is 2-cycloheptylpropionyl chloride. The molecular formula is as follows: c₁₉H₂₅BrN₂O; molecular weight: 377.3, respectively; mass/charge ratio: 376.1 (100.0%), 378.1 (99.7%), 377.1 (21.6%), 379.1 (21.2%), 380.1 (2.4%); elemental analysis: c, 60.48; h, 6.68; br, 21.18; n, 7.42; and O, 4.24.

Example 62

N- (4-chloro-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-chloroindole and Z is 2-cyclohexylacetyl chloride. The molecular formula is as follows: c₂₀H₂₈ClN₃O₂(ii) a Molecular weight: 377.9, respectively; mass/charge ratio: 377.2 (100.0%), 379.2 (34.9%), 378.2 (23.1%), 380.2 (7.7%); elemental analysis: c, 63.56; h, 7.47; cl, 9.38; n, 11.12; and O, 8.47.

Example 63

N- (4-bromo-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cyclohexylacetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-bromoindole and Z is 2-cyclohexylacetyl chloride. The molecular formula is as follows: c₂₀H₂₈BrN₃O₂(ii) a Molecular weight: 422.4, respectively; mass/charge ratio: 423.1 (100.0%), 421.1 (99.8%), 422.1 (23.1%), 424.1 (22.7%), 425.1 (2.9%); elemental analysis: c, 56.87; h, 6.68; br, 18.92; n, 9.95; and O, 7.58.

Example 64

N- (4-chloro-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-chloroindole and Z is 2-cycloheptylacetyl chloride. The molecular formula is as follows: c₂₁H₃₀ClN₃O₂(ii) a Molecular weight: 391.9, respectively; mass/charge ratio:391.2 (100.0%), 393.2 (35.2%), 392.2 (24.2%), 394.2 (8.1%), 395.2 (1.1%); elemental analysis: c, 64.35; h, 7.72; cl, 9.05; n, 10.72; and O, 8.16.

Example 65

N- (4-bromo-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cycloheptylacetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-bromoindole and Z is 2-cycloheptylacetyl chloride. The molecular formula is as follows: c₂₁H₃₀BrN₃O₂(ii) a Molecular weight: 436.4; mass/charge ratio: 437.2 (100.0%), 435.2 (99.5%), 436.2 (23.0%), 438.2 (22.7%), 439.2 (3.1%), 436.1 (1.1%), 438.1 (1.1%); elemental analysis: c, 57.80; h, 6.93; br, 18.31; n, 9.63; o, 7.33.

Example 66

2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) acetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-chloroindole and Z is bicyclo [2.2.2]Octane-1-yl acetyl chloride. The molecular formula is as follows: c₂₂H₃₀ClN₃O₂(ii) a Molecular weight: 403.9; mass/charge ratio: 403.2 (100.0%), 405.2 (35.4%), 404.2 (25.3%), 406.2 (8.4%), 407.2 (1.1%); elemental analysis: c, 65.41; h, 7.49; cl, 8.78; n, 10.40; and O, 7.92.

Example 67

2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) acetamide

Synthesized according to the method disclosed in example 23, wherein X is 4-bromoindole and Z is bicyclo [2.2.2]Octane-1-yl acetyl chloride. The molecular formula is as follows: c₂₂H₃₀BrN₃O₂(ii) a Molecular weight: 448.4, respectively; mass/charge ratio: 449.2 (100.0%), 447.2 (99.2%), 448.2 (24.0%), 450.2 (23.7%), 451.2 (3.4%), 448.1 (1.1%), 450.1 (1.1%); elemental analysis: c, 58.93; h, 6.74; br, 17.82; n, 9.37; and O, 7.14.

Example 68

Antagonism of P2X7R activity by N-indol-3-yl-acetamides and N-azaindol-3-yl-acetamides

Inhibition of P2X7R activity by the compounds of the invention was assessed by measuring calcium influx in Hek293 cells (ECACC No. 85120602) that have been stably transfected with cDNA of human P2X 7R.

Hek293 cells are human embryonic kidney cells which do not express endogenous P2X7R (Surprenant et al (1996) Science 272: 735-738). P2X7R expressing Hek293 cells were generated by lipofection of human P2X7R cDNA (genbank accession number BC011913) under the control of the human Cytomegalovirus (CMV) immediate early (immedate-early) promoter and inserted into pcdna3.1 vector (Invitrogen). At 37 ℃ with 8.5% CO₂Next, the cells were cultured in Darber's modified eagle medium (DMEM; GibcoBRL/Invitrogen) supplemented with heat-inactivated fetal bovine serum (10% v/v), 2mM L-glutamine, 100 units/ml penicillin, 0.1mg/ml streptomycin, and 750. mu.g/ml Genetiin G418 (GibcoBRL/Invitrogen).

The test compounds were monitored for Bz-ATP-stimulated P2X7 by detecting changes in calcium influx using Fluo-4-AM fluorescent dye according to the manufacturer's recommendations (Molecular Devices Corporation, U.S.A.)And (3) inhibition of R. Briefly, P2X 7R-expressing Hek293 cells were cultured in 96-well plates at a final density of about 10,000 cells per well. On the day of the experiment, the medium was completely removed from the wells and the cells were washed once with assay buffer (1X 20mM Hepes buffer, pH7.4 and 250mM probenecid in Hank's Balanced salt (HBSS); GibcoBRL/Invitrogen). Cells were incubated in 50. mu.l of assay buffer containing 100. mu.M Fluo-4AM fluorochrome per well for 1 hour at room temperature. The assay buffer containing Fluo-4AM fluorochrome was then removed, the cells were washed once with assay buffer (without Fluo-4AM), and then 100. mu.l of assay buffer (without Fluo-4AM) containing the test compound was added to each well. After 15 min incubation, 100 μ M Bz-ATP was added and fluorescence was detected in FlexStation II (Molecular Devices, U.S.A.) according to the following parameters: 485nm excitation wavelength; an emission wavelength of 525 nm; 515nm emission cut-off wavelength; 100 μ l pipette height; transfer volume of 25 μ l; 5 times the compound concentration; addition rate of 3. Test compounds were added at concentrations ranging from 0.001. mu.M up to 60. mu.M. Fluorescence data was processed with a delay time of 15 seconds, recording of 45 seconds, zero reference calibrated with 2 points, and% baseline multiplier set to 3. The resulting curve area was then calculated and half maximal Inhibitory Concentration (IC) for each test compound was determined using SoftMax Pro software (Molecular Devices, u.s.a.)₅₀). The compounds of the present invention inhibit P2X7R activity with an IC50 of 1 μ M to 0.001 μ M. For example, IC of the Compound described in example 2₅₀About 0.0038. mu.M.

Example 69

N-indol-3-yl-acetamides and N-azaindol-3-yl-acetamides compounds for reducing interleukin-1 beta secretion

Isolated human monocytes were used to evaluate the effect of the compounds of the invention on IL-1 β secretion.

Briefly, human monocytes were purified from human blood by Ficoll-Paque from the buffy coat as follows. The buffy coat is the grey white layer of leukocytes and platelets that accumulate on the surface of settled red blood cells when the blood is allowed to stand or centrifuged. All the white film layers (one for each donor)Aliquots) were diluted with PBS and 20ml were added on top of 15ml Ficoll-Paque. The slants were centrifuged at 900g for 20 min at room temperature. The white interface was transferred to a new tube, washed 3 times with PBS and subjected to 3 centrifugation steps (600, 400, 250g) at room temperature for 10 minutes each. Resuspending a cell pellet of Peripheral Blood Mononuclear Cells (PBMC) (1X 10)⁷cells/mL) in RPMI1640 supplemented with 5% heat-inactivated human serum. The PBMC suspension formed contained monocytes and lymphocytes. Monocytes were incubated at 37 ℃ with 5% CO₂After 24 hours of adhesion, the non-adhered lymphocytes were washed out with PBS. PBMC were incubated at 37 ℃ with 5% CO₂Upon incubation, macrophages differentiated within 5 days. On day 5, cells were counted at 1 × 10⁶The concentration of/ml was resuspended in RPMI1640 supplemented with 5% human serum and then placed in 24-well plates (5X 10)⁵Cells/well). On day 6, the medium was removed and replaced with RPMI1640 supplemented with 10% heat-inactivated fetal bovine serum to avoid IL-1 β contamination from human serum.

On the day of the experiment, macrophages were pre-stimulated with 1 μ g/mL LPS for 2 hours at 37 ℃. Then 100. mu.M BzATP was added to the cells and incubated at 37 ℃ for 30 minutes. AFC-5128 (concentration see figure) was added and 5 minutes later stimulated with BzATP. The control samples corresponded to cells that had not been subjected to any treatment. After incubation, the supernatant was collected by centrifugation (250g, 5 min) and assayed for IL-1 β secretion using the human IL-1 β/IL-1F2Quantikine ELISA Kit according to the manufacturer's instructions. Multiple donors were tested individually for AFC-5128. All treatments were tested in triplicate for each donor. For each data point, the o.d. at 450nm was determined and the concentration of IL-1 β was calculated based on a standard curve. IL1 β concentration was further calculated to have concentrations expressed in pg/mL/10E +6 cells and their standard deviation.

An example of a compound of the invention that reduces IL-1 β secretion (in this case, the compound described in example 2) is illustrated in FIG. 1.

Example 70

Analgesic and anti-inflammatory effects

This example illustrates the analgesic and anti-inflammatory benefits of the compounds of the present invention using a carrageenan-induced swelling inflammation model of the foot.

Adult male Sprague Dawley rats were stimulated by subcutaneous injection of carrageenan (1% suspension, 0.1ml) into the plantar side of the right hind paw of the rat. One hour after carrageenan challenge, a 0.5% methylcellulose suspension or vehicle (0.5% methylcellulose) of the compound was administered orally. The paw is then marked horizontally with washable ink on the lateral malleolus of the paw, so that the paw is immersed in the plethysmometer chamber until the mark. The plethysmometer allows detection of small volume changes of the paw. One hour after compound or vehicle administration (or 2 hours with carrageenan stimulation), the plantar foot test was performed and paw volumes were recorded.

For the plantar pain threshold test (plantar test), each rat was placed on a preheated glass stand. Both hind paws of the animals were stimulated with a radiant heat source. Withdrawal latencies of the paw to the stimulus were recorded. Prolongation of the response latency to the paw withdrawal is interpreted as an analgesic response. Three experiments were performed for each animal to obtain the average paw withdrawal latency. The mean withdrawal latencies (PWL) of the experimental groups were compared to the vehicle treated groups.

For the paw edema test, the increase in paw volume for each animal was calculated by subtracting the volume of the left hind paw from the volume of the right hind paw (paw volume difference-volume of right hind paw-volume of left hind paw). Inhibition of foot volume increase was interpreted as an anti-inflammatory response. The observed results were statistically validated using ANOVA Tukey's multiple comparison test. The results are shown in FIG. 2.

The compounds of the invention (compounds described in example 2) were evaluated for an increase in the withdrawal latency in response to a thermal stimulus, which is indicative of an analgesic response.

The inhibition of carrageenan-induced foot swelling, which is explained as an anti-inflammatory response, by the compounds of the present invention was also evaluated.

The accompanying drawings illustrate:

FIG. 1 inhibition of IL-1 beta secretion: (^＊p＜0.01)。

FIG. 2 analgesic and anti-inflammatory effects on inflammation model: (^＊＊＊p＜0.001)。

Claims (19)

1. A compound of general formula (I) or a pharmaceutically acceptable salt or solvate thereof:

wherein

-R₁Is mono-or dicycloalkylalkyl or mono-or dicycloalkyl;

-R₂selected from straight or branched C₁-C₅Alkyl, which may optionally be substituted byAnd (3) substitution: -OH, -CH₂-OH、C₁-C₅Alkoxy, NH₂-、N(R_a)₂-、NHR_a-、CN-、CF₃Halogen (i.e. Cl, F, Br or I), piperidin-1-yl, morpholin-4-yl, pyrrolidin-1-yl, 5H-tetrazolylpropyl, methylcarbamoyl, dimethylcarbamoyl or ethylmethylcarbamoyl, wherein R is_aIs C₁-C₅An alkyl group;

-R₃、R₄、R₅、R₆independently at each occurrence, selected from hydrogen, halogen (i.e., Cl, F, Br, or I), methyl, methoxy, cyano, or trifluoromethyl;

-a, b, c, d, x are independently at each occurrence selected from carbon or nitrogen.

2. The compound of claim 1, wherein R₂Substituted with one or two substituents selected from: -OH, -CH₂-OH、C₁-C₅Alkoxy, -NH₂、NHRa、-CN、-CF₃Halogen, piperidin-1-yl, morpholin-4-yl, pyrrolidin-1-yl, or 5H-tetrazolylpropyl.

3. The compound of claim 1 or 2, wherein R₁Is selected from cyclopentyl, cyclopentylmethyl, cyclohexyl, cyclohexylmethyl, cycloheptyl, cycloheptylmethyl, bicyclo [2.2.2]Octane-1-yl and bicyclo [2.2.2]Octane-1-ylmethyl.

4. The compound of claim 3, wherein R₂Is C₁-C₅Alkyl or C₂-C₅A hydroxyalkyl group.

5. The compound of any one of claims 1 to 4, wherein R₃、R₄、R₅And R₆At least two of which are hydrogen.

6. The compound of any one of claims 1 to 5, wherein x is CH.

7. The compound of any one of claims 1 to 5, wherein x is N.

8. The compound of any one of claims 1 to 7, wherein a, b, C and d are C.

9. The compound of any one of claims 1 to 7, wherein one of a, b, c and d is N.

10. The compound of any one of claims 1 to 9, selected from:

-N- (4-chloro-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-chloro-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1- (2-hydroxyethyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-chloro-1- (hydroxymethyl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-bromo-1- (hydroxymethyl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-chloro-1- (hydroxymethyl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-bromo-1- (hydroxymethyl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-chloro-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cyclohexylacetamide,

-N- (4-chloro-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide,

-N- (4-chloro-1- (2-hydroxypropyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (2-hydroxypropyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -2-cycloheptylacetamide,

-N- (4-chloro-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -3-cyclohexylpropionamide,

-N- (4-bromo-1- (2-hydroxyethyl) -1H-pyrrolo [2, 3-b ] pyridin-3-yl) -3-cyclohexylpropionamide,

-N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) acetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) acetamide,

-N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-chloro-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-bromo-1- (2-hydroxypropyl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) acetamide

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) acetamide,

-N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-chloro-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-bromo-1- (1, 3-dihydroxypropan-2-yl) -1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -2- (bicyclo [2.2.2] octan-1-yl) acetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -2- (bicyclo [2.2.2] octan-1-yl) acetamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-chloro-1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (1- (3-amino-2-hydroxypropyl) -4-bromo-1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-chloro-1-methyl-1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1-methyl-1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-chloro-1-methyl-1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1-methyl-1H-indol-3-yl) -2-cycloheptylacetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1-methyl-1H-indol-3-yl) acetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1-methyl-1H-indol-3-yl) acetamide,

-N- (4-chloro-1-methyl-1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-bromo-1-methyl-1H-indol-3-yl) -3-cyclohexylpropionamide,

-N- (4-chloro-1-methyl-1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-bromo-1-methyl-1H-indol-3-yl) -3-cycloheptylpropanamide,

-N- (4-chloro-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-bromo-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cyclohexylacetamide,

-N- (4-chloro-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-N- (4-bromo-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) -2-cycloheptylacetamide,

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-chloro-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) acetamide, and

-2- (bicyclo [2.2.2] octan-1-yl) -N- (4-bromo-1- (2-hydroxy-3- (methylamino) propyl) -1H-indol-3-yl) acetamide.

11. A pharmaceutical composition comprising a compound of any one of claims 1 to 10.

12. The pharmaceutical composition of claim 11, further comprising an additional active compound in separate or unit dosage forms for simultaneous or sequential administration.

13. A pharmaceutical composition according to claim 11 or 12 for use in the treatment of affective disorders.

14. The pharmaceutical composition according to claim 13, wherein the affective disorder is selected from the group consisting of depression, anxiety, bipolar disorder and schizophrenia.

15. The pharmaceutical composition of claim 11 or 12 for use in the treatment of neurodegenerative diseases and disorders, diseases and disorders mediated by or causing neuroinflammation, and centrally mediated neuropsychiatric diseases and disorders.

16. The pharmaceutical composition according to claim 11 or 12 for use in the treatment of pain, inflammatory processes and degenerative disorders.

17. The pharmaceutical composition of claim 16, wherein the inflammatory process is selected from the group consisting of rheumatoid arthritis, osteoporosis, and chronic obstructive pulmonary disease.

18. The pharmaceutical composition of claim 16, wherein the degenerative disorder is selected from the group consisting of glaucoma, age-related macular degeneration, uveitis, neuropathic pain, multiple sclerosis, amyotrophic lateral sclerosis, parkinson's disease, and alzheimer's disease.

19. A pharmaceutical composition according to claim 11 or 12 for use in the treatment of neuropathic pain.