Diaryl-substituted 6,5-fused ring compounds as C5a inhibitors

Abstract

The present invention particularly provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is a modulator of the C5a receptor. Also provided are pharmaceutical compositions and methods of use, including treatment of diseases or conditions involving pathological activation of C5a and non-pharmaceutical applications. #imgabs0#

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 609,834, filed on December 22, 2017, under 35 U.S.C. §119(e), which is incorporated herein by reference in its entirety.

[0003] STATEMENT AS TO RIGHTS TO INventions Made Under Federally Sponsored Research and Development

[0004] not applicable.

[0005] Reference to a "Sequence Listing", table or computer program listing appendix submitted on a CD-ROM

[0006] not applicable. Background of the Invention

[0008] The complement system plays a central role in the clearance of immune complexes and in the immune response to infectious agents, foreign antigens, virus-infected cells, and tumor cells. Inappropriate or excessive activation of the complement system can lead to harmful and potentially life-threatening consequences due to severe inflammation and resulting tissue destruction. These consequences are clinically manifested in a variety of diseases, including septic shock; myocardial and intestinal ischemic / reperfusion injury; transplant rejection; organ failure; nephritis; pathological inflammation and autoimmune diseases.

[0009] The complement system consists of a group of proteins that are usually present in the serum in an inactive state. The activation of the complement system mainly includes three different pathways, namely the classical pathway, the alternative pathway and the lectin pathway (VM Holers, In Clinical Immunology: Principles and Practice, RR Rich, Mosby Press; 1996, 363-391): 1) The classical pathway is a calcium / magnesium-dependent cascade, which is usually activated by the formation of an antigen-antibody complex. It can also be activated in an antibody-independent manner by binding to C-reactive protein (complexed with ligand) complexed with a ligand, and by many pathogens (including Gram-negative bacteria). 2) The alternative pathway is a magnesium-dependent cascade, which is activated by the deposition and activation of C3 on certain sensitive surfaces (such as cell wall polysaccharides of yeast and bacteria, and certain biopolymer materials). 3) The lectin pathway involves the initial binding of mannose-binding lectin and subsequent activation of C2 and C4, which is common to the classical pathway (Matsushita, M et al., J. Exp. Med. 176: 1497-1502 (1992); Suankratay, C et al., J. Immunol 160: 3006-3013 (1998)).

[0010] Activation of the complement pathway generates biologically active fragments of complement proteins, such as C3a, C4a, and C5a anaphylatoxins and the C5b-9 membrane attack complex (MAC), which mediate inflammatory responses by affecting leukocyte chemotaxis; activating macrophages, neutrophils, platelets, mast cells, and endothelial cells; and increasing vascular permeability, cell lysis, and tissue damage.

[0011] Complement C5a is one of the most potent pro-inflammatory mediators of the complement system. (In terms of causing inflammatory responses, the allergic C5a peptide is 100 times more potent than C3a on a molar basis.) C5a is the activated form of C5 (190 kD, molecular weight). C5a is present in human serum at about 80 μg / ml (Kohler, PF et al., Journal of Immunology (J.Immunol) 99:1211-1216 (1967)). It consists of two polypeptide chains (α and β), with molecular weights of approximately 115 kD and 75 kD, respectively (Tack, BF et al., Biochemistry (Biochemistry) 18:1490-1497 (1979)). Biosynthesis is a single-chain promolecule, and during processing and secretion, C5 is enzymatically cleaved into a double-chain structure. After cleavage, the two chains are held together by at least one disulfide bond as well as non-covalent interactions (Ooi, YM et al., J. Immunol 124:2494-2498 (1980)).

[0012] During the activation of the complement pathway, C5 is cleaved into C5a and C5b fragments. The convertase responsible for C5 activation is a multi-subunit complex of C4b, C2a and C3b for the classical pathway, and a multi-subunit complex of (C3b)2, Bb and P for the alternative pathway (Goldlust, MB et al., J. Immunol. 113:998-1007 (1974); Schreiber, RD et al., Proc. Natl. Acad. Sci. 75:3948-3952 (1978)). In the α-chain, C5 is activated by cleavage at positions 74-75 (Arg-Leu). After activation, the 11.2 kD, 74 amino acid peptide C5a from the amino terminal portion of the α-chain is released. Both C5a and C3a are potent stimulators of neutrophils and monocytes (Schindler, R et al., Blood 76:1631-1638 (1990); Haeffner-Cavaillon, N et al., J. Immunol 138:794-700 (1987); Cavaillon, JM et al., Eur. J. Immunol 20:253-257 (1990)).

[0013] In addition to its anaphylactic toxicity, C5a also induces chemotactic migration of neutrophils (Ward, PA et al., J. Immunol 102:93-99 (1969)), eosinophils (Kay, AB et al., Immunol 24:969-976 (1973)), basophils (Lett-Brown, MA et al., J. Immunol 117:246-252 1976)) and monocytes (Snyderman, R et al., Proc. Soc. Exp. Biol. Med. 138:387-390 1971)). Both C5a and C5b-9 activate endothelial cells to express adhesion molecules that are critical for sequestering activated leukocytes, which mediate tissue inflammation and damage (Foreman, KE et al., J. Clin. Invest. 94: 1147-1155 (1994); Foreman, KE et al., Inflammation 20: 1-9 (1996); Rollins, SA et al., Transplantation 69: 1959-1967 (2000)). C5a also mediates inflammatory responses by causing smooth muscle contraction, increasing vascular permeability, inducing basophil and mast cell degranulation, and inducing the release of lysosomal proteases and oxidative free radicals (Gerard, C et al., Ann. Rev. Immunol. 12: 775-808 (1994)). In addition, C5a modulates hepatic acute phase gene expression and enhances overall immune responses by increasing production of TNF-α, IL-1-β, IL-6, IL-8, prostaglandins, and leukotrienes (Lambris, JD et al., In: The Human Complement System in Health and Disease, Volanakis, JE ed., Marcel Dekker, New York, NY 83-118).

[0014] The allergic reaction and chemotaxis of C5a are believed to be mediated by its interaction with the C5a receptor. The human C5a receptor (C5aR) is a 52kD membrane-bound G protein-coupled receptor expressed in neutrophils, monocytes, basophils, eosinophils, hepatocytes, pulmonary smooth muscle and endothelial cells, and glomerular tissue (Van-Epps, DE et al., Journal of Immunology (J. Immunol.) 132: 2862-2867 (1984); Haviland, DL et al., Journal of Immunology (J. Immunol.) 154: 1861-1869 (1995); Wetsel, RA, Immunol. Leff. 44: 183-187 (1995); Buchner, RR et al., J. Immunol. 155: 308-315 (1995); Chenoweth, DE et al., Proc. Natl. Acad. Sci., 75: 3943-3947 (1978); Zwirner, J et al., Mol. Immunol., 36: 877-884 (1999). The ligand binding site of C5aR is complex and consists of at least two physically separable binding domains. One binds to the C5a amino terminus (amino acids 1-20) and the disulfide-linked core (amino acids 21-61), while the other binds to the C5a carboxyl terminus (amino acids 62-74) (Wetsel, RA, Curr. Opin. Immunol. 7:48-53 (1995)).

[0015] C5a plays an important role in inflammation and tissue damage. In cardiopulmonary bypass and hemodialysis, when human blood comes into contact with the artificial surfaces of a heart-lung machine or a kidney dialysis machine, the formation of C5a is the result of activation of the bypass complement pathway (Howard, RJ et al., Arch. Surg. 123:1496-1501 (1988); Kirklin, JK et al., J. Cardiovasc. Surg. 86:845-857 (1983); Craddock, PR et al., N. Engl. J. Med. 296:769-774 (1977)). C5a causes capillary permeability and edema, bronchoconstriction, pulmonary vasoconstriction, leukocyte and platelet activation, and increased infiltration into tissues, particularly the lungs (Czermak, BJ et al., J. Leukoc. Biol. 64:40-48 (1998)). Administration of anti-C5a monoclonal antibodies has been shown to reduce coronary endothelial dysfunction caused by cardiopulmonary bypass and cardioplegia (Tofukuji, M et al., J. Thorac. Cardiovasc. Surg. 116:1060-1068 (1998)).

[0016] C5a is also involved in acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD) and multiple organ failure (MOF) (Hack, CE et al., Am. J. Med. 1989: 86: 20-26; Hammerschmidt DE et al., Lancet 1980; 1: 947-949; Heideman M. et al., J. Trauma 1984; 4: 1038-1043; Marc, MM, et al., Am. J. Respir. Cell and Mol. Biol., 2004: 31: 216-219). C5a increases the production of two important proinflammatory cytokines (TNF-α and IL-1) by monocytes. In animal models of septic shock, C5a has also been shown to play an important role in the progression of tissue damage, especially lung damage (Smedegard G et al., Am. J. Pathol., 1989; 135: 489-497; Markus, S et al., FASEB Journal (2001), 15: 568-570). In sepsis models using rats, pigs, and non-human primates, administration of anti-C5a antibodies to animals prior to endotoxin or E. coli treatment reduced tissue damage and IL-6 production (Smedegard, G et al., Am. J. Pathol. 135:489-497 (1989); Hopken, U et al., Eur. J. Immunol. 26:1103-1109 (1996); Stevens, JH et al., J. Clin. Invest. 77:1812-1816 (1986)). More importantly, blocking C5a or C5a with anti-C5a polyclonal antibodies was shown to significantly improve survival in a rat cecal ligation / puncture model of sepsis. (Czermak, BJ et al., Nat. Med. 5:788-792 (1999)). This model has many similarities to the clinical manifestations of sepsis in humans. (Parker, SJ et al., Br. J. Surg. 88:22-30 (2001)). In the same sepsis model, anti-C5a antibodies have been shown to inhibit thymocyte apoptosis (Guo, RF et al., J. Clin. Invest. 106:1271-1280 (2000)) and prevent MOF (Huber-Lang, M et al., J. Immunol. 166:1193-1199 (2001)).Anti-C5a antibodies also have a protective effect in the cobra venom factor model of lung injury in rats, as well as in immune complex-induced lung injury (Mulligan, MS et al., J. Clin. Invest. 98:503-512 (1996)). The importance of C5a in immune complex-mediated lung injury was later demonstrated in mice (Bozic, CR et al., Science 26:1103-1109 (1996)).

[0017] C5a was found to be a major mediator of myocardial ischemia-reperfusion injury. Complement depletion reduced myocardial infarct size in mice (Weisman, HF et al., Science 249: 146-151 (1990)), and treatment with anti-C5a antibodies reduced injury in the rat hindlimb ischemia-reperfusion model (Bless, NM et al., Am. J. Physiol. 276: L57-L63 (1999)). Reperfusion injury during myocardial infarction was also significantly reduced in pigs treated with monoclonal anti-C5a IgG (Amsterdam, EA et al., Am. J. Physiol. 268: H448-H457 (1995)). A recombinant human C5aR antagonist reduced infarct size in a porcine model of surgical revascularization (Riley, RD et al., J. Thorac. Cardiovasc. Surg. 120:350-358 (2000)).

[0018] The neutrophil driven by C5a also causes many bullous diseases (such as bullous pemphigus, pemphigus vulgaris and pemphigus foliaceus). These are chronic and recurrent inflammatory diseases, characterized by the sterile blisters that occur in the subepidermal space of the skin and mucous membranes clinically. Although it is believed that the autoantibodies to the keratinocytes located at the basement membrane of the skin are the basis for the separation of epidermal basal keratinocytes from the basement membrane below, another feature of the blisters is the accumulation of neutrophils in the upper dermis and the blister cavity. In experimental models, even in the presence of high autoantibody titers, the reduction of neutrophils or the lack of complement (total or C5 selective) can also inhibit the formation of subepidermal blisters.

[0019] Complement levels are elevated in patients with rheumatoid arthritis (Jose, PJ et al., Ann. Rheum. Dis. 49:747-752 (1990); Grant, EP et al., J. of Exp. Med., 196(11):1461-1471, (2002)), lupus nephritis (Bao, L., et al., Eur. J. of Immunol., 35(8), 2496-2506, (2005)) and systemic lupus erythematosus (SLE) (Porcel, JM et al., Clin. Immunol. Immunopathol. 74:283-288 (1995)). C5a levels correlate with the severity of the disease state. Collagen-induced arthritis in mice and rats resembles rheumatoid arthritis disease in humans. Mice lacking the C5a receptor show complete protection against arthritis induced by injection of monoclonal anti-collagen Abs (Banda, NK et al., J. of Immunol., 2003, 171: 2109-2115). Therefore, inhibition of C5a and / or C5a receptor (C5aR) may be useful in treating these chronic diseases.

[0020] It is believed that the complement system is activated in patients with inflammatory bowel disease (IBD) and is thought to play a role in the pathogenesis of the disease. Activated complement products have been found on the luminal surface of surface epithelial cells of IBD patients and in the muscularis mucosa and submucosal blood vessels (Woodruff, TM et al., J of Immunol., 2003, 171: 5514-5520).

[0021] C5aR expression is upregulated in reactive astrocytes, microglia, and endothelial cells in the inflamed human central nervous system (Gasque, P et al., Am. J. Pathol. 150:31-41 (1997)). C5a may be involved in neurodegenerative diseases such as Alzheimer's disease (Mukherjee, P et al., J. Neuroimmunol. 105:124-130 (2000); O'Barr, S et al., J. Neuroimmunol., (2000) 105:87-94; Farkas, I et al., J. Immunol., (2003) 170:5764-5771), Parkinson's disease, Pick's disease, and transmissible spongiform encephalopathy. Activation of neuronal C5aR can induce apoptosis (Farkas I et al., J. Physiol. 1998; 507: 679-687). Therefore, inhibition of C5a and / or C5aR can also be used to treat neurodegenerative diseases.

[0022] There is evidence that C5a production exacerbates inflammation associated with atopic dermatitis (Neuber, K et al., Immunology 73:83-87, (1991)) and chronic urticaria (Kaplan, AP, J. Allergy Clin. Immunol. 114; 465-474, (2004)).

[0023] It is now known that psoriasis is a T-cell mediated disease (Gottlieb, EL et al., Nat. Med. 1:442-447 (1995)). However, neutrophils and mast cells may also be involved in the pathogenesis of the disease (Terui, T et al., Exp. Dermatol. 9:1-10; 2000); Werfel, T et al., Arch. Dermatol. Res. 289:83-86 (1997)). In highly inflamed areas of psoriatic plaques, accumulation of neutrophils under the stratum corneum has been observed, and extracts of psoriasis lesions (scaly) contain highly elevated levels of C5a and show potent chemotactic activity for neutrophils (an effect that can be inhibited by the addition of C5a antibodies). T cells and neutrophils can be chemoattracted by C5a (Nataf, S. et al., J. Immunol. 162:4018-4023 (1999); Tsuji, RF et al., J. Immunol. 165:1588-1598 (2000); Cavaillon, JM et al., Eur. J. Immunol. 20:253-257 (1990)). In addition, the expression of C5aR has been demonstrated in plasmacytoid dendritic cells (pDC) isolated from lesions of cutaneous lupus erythematosus, and these cells show chemotactic behavior to C5a, suggesting that blocking C5aR on pDC may be effective in reducing pDC infiltration of SLE and psoriasis skin. Therefore, C5a may be an important therapeutic target for the treatment of psoriasis.

[0024] Immune complexes (ICs) containing immunoglobulin G contribute to the pathophysiology of many autoimmune diseases, such as systemic lupus erythematosus, rheumatoid arthritis, Sjogren's disease, Goodpasture's syndrome, and hypersensitivity pneumonitis (Madaio, MP, Semin. Nephrol. 19:48-56 (1999); Korganow, AS et al., Immunity 10:451-459 (1999); Bolten, WK, Kidney Int. 50:1754-1760 (1996); Ando, ​​M. et al., Curr. Opin. Pulm. Med. 3:391-399 (1997)). These diseases are highly heterogeneous and usually affect one or more of the following organs: skin, blood vessels, joints, kidneys, heart, lungs, nervous system and liver (including cirrhosis and liver fibrosis). The classic animal model for inflammatory response in these IC diseases is the Arthus reaction, which is characterized by polymorphonuclear cell infiltration, hemorrhage and plasma exudation (Arthus, M., CR Soc. Biol. 55: 817-824 (1903)). Recent studies have shown that C5aR-deficient mice are protected from IC-induced tissue damage (Kohl, J et al., Mol. Immunol. 36: 893-903 (1999); Baumann, U et al., J. Immunol. 164: 1065-1070 (2000)). This result is consistent with the observation that a small peptide anti-C5aR antagonist inhibited the inflammatory response caused by IC deposition (Strachan, AJ et al., J. Immunol. 164: 6560-6565 (2000)). C5a (together with its receptor) plays an important role in the pathogenesis of IC diseases. C5a and C5aR inhibitors can be used to treat these diseases.

[0025] Related technical description :

[0026] Non-peptide based C5a receptor antagonists have been reported to be effective in treating endotoxin shock in rats (Stracham, AJ et al., J. of Immunol. (2000), 164(12):6560-6565); and for treating IBD in a rat model (Woodruff, TM et al., J. of Immunol., 2003, 171:5514-5520). Non-peptide based C5a receptor modulators have also been described in the patent literature of Neurogen Corporation (e.g. WO2004 / 043925, WO2004 / 018460, WO2005 / 007087, WO03 / 082826, WO03 / 08828, WO02 / 49993, WO03 / 084524); Dompe SPA (WO02 / 029187); University of Queensland (WO2004 / 100975); and ChemoCentryx (WO2010 / 075257).

[0027] There is a wealth of experimental evidence in the literature that elevated levels of C5a are associated with a variety of diseases and conditions, particularly autoimmune and inflammatory diseases and conditions. Thus, there remains a need in the art for new organic small molecule C5a receptor (C5aR) modulators (e.g., agonists, preferably antagonists, partial agonists) that can be used to inhibit pathogenic events, such as chemotaxis associated with anaphylatoxin activity. The present invention meets this need and other needs. Summary of the invention

[0028] In one aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof:

[0029]

[0030] Among them, symbols, letters and subscripts n, m, a, b, e, X 1 , R 1 , R 2a , R 2b , R 3 , R 4 and R 5 has the meaning described below.

[0031] In addition to the compounds provided herein, the present invention also provides pharmaceutical compositions containing one or more of these compounds, as well as the use of these compounds in methods of treatment, primarily for the treatment of diseases associated with C5a signaling activity.

[0032] In other aspects, the present invention provides methods for diagnosing diseases in individuals. In these methods, the compounds provided herein are administered to the subject in a labeled form, followed by diagnostic imaging to determine the presence or absence of C5aR and / or the location of cells expressing C5aR receptors. In related aspects, the method for diagnosing a disease is performed by contacting a tissue or blood sample with a labeled compound provided herein and determining the presence, absence, amount or location of C5aR in the sample.

[0033] BRIEF DESCRIPTION OF THE DRAWINGS

[0034] not applicable. DETAILED DESCRIPTION OF THE INVENTION

[0036] I. Abbreviations and Definitions

[0037] Unless otherwise indicated, the term "alkyl", by itself or as part of another substituent, refers to a radical having the specified number of carbon atoms (i.e., C 1-8 Refers to a straight or branched hydrocarbon group of 1 to 8 carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and similar groups. The term "alkenyl" refers to an unsaturated alkyl group having one or more double bonds. Similarly, the term "alkynyl" refers to an unsaturated alkyl group having one or more triple bonds. Examples of such unsaturated alkyl groups include: vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), isobutenyl, 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologues and isomers. The term "cycloalkyl" refers to a ring having a specified number of atoms (e.g., C 3-6 "Cycloalkyl" may also refer to bicyclic hydrocarbon rings and polycyclic hydrocarbon rings, for example, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term "heterocycloalkyl" refers to a cycloalkyl group containing one to five heteroatoms selected from N, O and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. The heterocycloalkyl group may be a monocyclic, bicyclic or polycyclic ring system. Non-limiting examples of heterocycloalkyl include: pyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, and similar groups. Heterocycloalkyl can be connected to the rest of the molecule through a ring carbon or a heteroatom.

[0038] The term "alkylene" by itself or as part of another substituent refers to a divalent group derived from an alkane, such as -CH2CH2CH2CH2-. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with those groups having 10 or fewer carbon atoms being preferred in the present invention. A "lower alkyl" or "lower alkylene" is a short chain alkyl or alkylene group typically having four or fewer carbon atoms. Similarly, "alkenylene" and "alkynylene" refer to unsaturated forms of "alkylene" having double or triple bonds, respectively.

[0039] Unless otherwise stated, the term "heteroalkyl" by itself or in combination with another term refers to a stable straight or branched or cyclic hydrocarbon group, or a combination thereof, consisting of the stated number of carbon atoms and one to three heteroatoms selected from O, N, Si and S, and wherein the nitrogen and sulfur atoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized. The heteroatoms O, N and S may be located at any position within the heteroalkyl group. The heteroatom Si may be located at any position of the heteroalkyl group, including the position where the alkyl group is attached to the rest of the molecule. Examples include: -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2, -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3 and -CH=CH-N(CH3)-CH3. Up to two heteroatoms may be consecutive, e.g., -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. Similarly, unless otherwise indicated, the terms "heteroalkenyl" and "heteroalkynyl" by themselves or in combination with another term, mean an alkenyl or alkynyl group, respectively, containing the stated number of carbon atoms and having one to three heteroatoms selected from O, N, Si and S, and wherein the nitrogen and sulfur atoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized. The heteroatoms O, N and S may be located at any position within the heteroalkyl group.

[0040] The term "heteroalkylene" by itself or as part of another substituent refers to a saturated or unsaturated or polyunsaturated divalent radical derived from a heteroalkyl group, for example, -CH2-CH2-S-CH2CH2- and -CH2-S-CH2-CH2-NH-CH2-, -O-CH2-CH=CH-, -CH2-CH=C(H)CH2-O-CH2- and -S-CH2-C≡C-. For heteroalkylene, heteroatoms can also be located at one or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, etc.).

[0041] The terms "alkoxy", "alkylamino" and "alkylthio" (or thioalkoxy) are used in their conventional sense to refer to those alkyl groups attached to the rest of the molecule through an oxygen atom, an amino group or a sulfur atom, respectively. Additionally, for dialkylamino groups, the alkyl moieties may be the same or different and may be combined with the nitrogen atom to which they are attached to form a 3-7 membered ring. Thus, with -NR a R b The groups represented are intended to include piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl, and the like.

[0042] The term "hydroxyalkyl" is used in its conventional sense to refer to a branched or straight chain alkyl group substituted with at least one hydroxy group. The hydroxy group may be at any position in the alkyl group. For example, the term "C 1-4 The term "hydroxyalkyl" includes hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxyisopropyl, and the like.

[0043] Unless otherwise stated, the terms "halo" or "halogen," by themselves or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as "haloalkyl" are intended to include monohaloalkyl and polyhaloalkyl. For example, the term "C 1-4 "Haloalkyl" is intended to include: trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0044] Unless otherwise indicated, the term "aryl" refers to a polyunsaturated, usually aromatic hydrocarbon group, which can be a monocyclic or polycyclic (up to three rings) fused together or covalently linked. The term "heteroaryl" refers to an aryl group (or ring) containing one to five heteroatoms selected from N, O and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom is optionally quaternized. The heteroaryl group can be connected to the rest of the molecule through a heteroatom. Non-limiting examples of aryl include: phenyl, naphthyl and biphenyl; and non-limiting examples of heteroaryl include: pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl, benzooxazolyl, benzotriazolyl, benzisoxazolyl, isobenzofuranyl, isoindolyl , indolizinyl, benzotriazinyl, thienopyridinyl, thienopyrimidinyl, pyrazolopyrimidinyl, pyrrolopyridinyl, imidazopyridinyl, benzothiaxolyl, benzofuranyl, benzothiophenyl, indolyl, quinolyl, isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furanyl, thienyl, and the like. Substituents for each of the above-recited aryl and heteroaryl ring systems are selected from the group consisting of acceptable substituents described hereinafter.

[0045] The term "pharmaceutically acceptable salts" is meant to include salts of the active compounds prepared with relatively nontoxic acids or bases, depending on the particular substituents present on the compounds described herein. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral forms of these compounds with a sufficient amount of the desired base, either in the absence of a solvent or in a suitable inert solvent. Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, trivalent manganic salts, divalent manganic salts, potassium salts, sodium salts, zinc salts, and the like. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary and tertiary amines, including substituted amines, cyclic amines, naturally occurring amines, and the like, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral forms of these compounds with a sufficient amount of the desired acid, which can be solvent-free or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, and salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. Also included are salts of amino acids such as arginine, and organic acids such as glucuronic acid or galacturonic acid (see, e.g., Berge, SM et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present invention contain both basic and acidic functional groups, which can be converted into base or acid addition salts.

[0046] The neutral forms of the compounds can be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from its various salt forms in certain physical properties, such as solubility in polar solvents, but for the purposes of the present invention the salts are otherwise equivalent to the parent form of the compound.

[0047] In addition to salt forms, the present invention provides compounds in the form of prodrugs. Prodrugs of compounds described herein are those compounds that are susceptible to chemical changes under physiological conditions to provide compounds of the present invention. In addition, prodrugs can be converted into compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, when the prodrug is placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent, the prodrug will slowly convert into the compound of the present invention.

[0048] Some compounds of the present invention can exist in unsolvated form as well as solvated form, including hydrated form. Generally, solvated form is comparable to unsolvated form, and is intended to be included within the scope of the present invention. Some compounds of the present invention can exist in polycrystalline or amorphous form. Generally, all physical forms are equivalent for the purposes contemplated by the present invention, and are intended to be within the scope of the present invention.

[0049] Certain compounds of the invention have asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, regioisomers, and individual isomers (e.g., individual enantiomers) are intended to be included within the scope of the invention. The compounds of the invention may also contain unnatural proportions of atomic isotopes at one or more of the atoms that make up these compounds. For example, the compounds may be substituted with radioactive isotopes such as tritium ( 3 H), iodine-125 ( 125 I) or carbon-14 ( 14 C)) is radiolabeled. All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.

[0050] As used herein, a wavy line that intersects a single, double, or triple bond in any chemical structure described herein is Indicates the point of attachment of a single, double, or triple bond to the rest of the molecule.

[0051] II. Specific implementation methods

[0052] A. Compounds

[0053] In one aspect, the present invention provides a compound of formula (I):

[0054]

[0055] or a pharmaceutically acceptable salt thereof, wherein

[0056] The ring vertex a is N or C (R 2c ), the ring vertex b is N or C (R 2d ) and the ring vertex e is N or C(R 2e ), where at most one of a, b and e is N;

[0057] X 1 Selected from the group consisting of: key, C 1-8 Alkylene, C(O), C(O)-C 1-4 Alkylene and S(O)2;

[0058] R 1 Select from the following group:

[0059] a) a 5- to 10-membered heteroaryl group having 1 to 4 heteroatoms selected from N, O and S as ring vertices;

[0060] b)C 6-10 Aryl;

[0061] c)C 3-8 Cycloalkyl;

[0062] d) 4- to 8-membered heterocycloalkyl having 1 to 2 heteroatoms selected from N, O and S as ring vertices;

[0063] e)C 1-8 Alkyl, C 1-8 Alkoxy, C 1-8 Haloalkyl, –C(O)NR 1a R 1b and –CO2R 1a ; Among them, R 1a and R 1b are independently selected from the group consisting of hydrogen, C 1-8 Alkyl, C 6-10 Aryl and –C 1-6 Alkylene-C 6-10 Aryl;

[0064] Among them, the group –X 1 -R 1 Optionally 1 to 5 R x Substituted by a substituent;

[0065] R 2a and R 2e are independently selected from the group consisting of hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, -OC 1-6 Halogenated alkyl, -SC 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6 Alkyl, -C 1-6 Alkyl-SC 1-6 alkyl, CN and halogen; and R 2a and R 2e At least one of them is not hydrogen;

[0066] R 2b, R 2c and R 2d are independently selected from the group consisting of hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, -OC 1-6 Halogenated alkyl, -SC 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6 Alkyl, -C 1-6 Alkyl-SC 1-6 Alkyl, cyano and halogen;

[0067] Each R 3 Independently selected from the group consisting of hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 and optionally two R on the same carbon atom 3 The groups are combined to form an oxo group (=O), and optionally two R 3 The group and the carbon atom to which it is attached form a 3-6 membered ring having 0-2 heteroatoms selected from O, N and S as ring vertices;

[0068] R 4 Independently selected from the group consisting of: –X 2 -OR 4a ,–X 2 -NR 4a R 4b ,–X 2 -CONR 4a R 4b ,–X 2 -NR 4a -C(O)R 4a , -X 2 -NR 4a -C(O)NR 4a R 4b , -X 2 -NR 4a -C(O)OR 4a , -X 2 -NR 4a -C(O)-C 1-3 Alkylene-OR 4a and –X 2 -NR 4a -C(O)-C 1-3 Alkylene-NR 4a R 4b ; Among them, each X 2 are independently a bond, C(O), C 1-4 Alkylene, C(O)-C 1-4 Alkylene and C1-4 Alkylene-C(O); and each R 4a and R 4b Independently selected from the group consisting of hydrogen, C 1-4 Alkyl and C 1-4 Haloalkyl;

[0069] Each R 5 Independently selected from the following group: C 1-8 Alkyl, C 1-8 Alkoxy, C 1-8 Haloalkyl, C 1-8 Halogenated alkoxy, C 1-8 Hydroxyalkyl, halogen, OH, CN, C(O)R 5a and CO2R 5a ; Among them, each R 5a Independently selected from the group consisting of hydrogen, C 1-4 Alkyl and C 1-4 Haloalkyl;

[0070] Each R x Independently selected from the group consisting of halogen, CN, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyl, C 2-4 Alkenyl, C 3-6 Cycloalkyl, CO2-C 1-4 Alkyl and CONH2;

[0071] subscript m is 0, 1, 2, 3, or 4; and

[0072] Subscript n is 0, 1, 2 or 3.

[0073] In one group of embodiments of the compounds of formula (I), R 4 Select from the following group:

[0074]

[0075] In another group of embodiments of the compounds of formula (I), or pharmaceutically acceptable salts thereof, R 4 Select from the following group:

[0076]

[0077] In another group of embodiments of the compounds of formula (I), or pharmaceutically acceptable salts thereof, wherein R 4 Select from the following group:

[0078]

[0079] With respect to the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any of the above groups of embodiments, in certain selected embodiments, X 1 is a bond; in other selected embodiments, X 1 is C(O); in another selected embodiment, X 1 C 1-8 In another selected embodiment, X 1 C(O)-C 1-4 Alkylene or S(O)2.

[0080] Regarding the compound of formula (I) or a pharmaceutically acceptable salt thereof, and any group or selected embodiment described above, in some further embodiments, wherein R 1 is a 5- to 10-membered heteroaryl group having 1 to 4 heteroatoms selected from N, O and S as ring vertices; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x In a further embodiment, R 1 is selected from the group consisting of pyrazolyl, pyridinyl, pyrimidinyl, imidazolyl, thiazolyl, thiadiazolyl and pyrazinyl; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

[0081] Regarding the compound of formula (I) or a pharmaceutically acceptable salt thereof, and any group or selected embodiment described above, in some further embodiments, wherein R 1 C 6-10 Aryl; and wherein the group –X 1 -R 1 Optionally 1 to 4 R x In yet further embodiments, R 1 is phenyl; and wherein the group –X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

[0082] Regarding the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any group or selected embodiment described above, in some further embodiments, R 1 C 3-8 Cycloalkyl; and wherein the group –X 1 -R 1 Optionally 1 to 4 R x In yet further embodiments, R 1is selected from the group consisting of cyclobutyl, cyclopentyl and cyclohexyl; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

[0083] Regarding the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any group or selected embodiment described above, in some further embodiments, R 1 is a 4- to 8-membered heterocycloalkyl group having 1 to 2 heteroatoms selected from N, O and S as ring vertices; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x In a further selected embodiment, R 1 is selected from the group consisting of oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, and morpholinyl; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

[0084] With respect to the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any group or selected embodiment described above, in some further embodiments, R 1 Select from the following group: C 1-8 Alkyl, C 1-8 Alkoxy, C 1-8 Haloalkyl, –C(O)NR 1a R 1b and –CO2R 1a ; Among them, R 1a and R 1b are independently selected from the group consisting of hydrogen, C 1-8 Alkyl, C 6-10 Aryl and –C 1-6 Alkylene-C 6-10 Aryl; and wherein the group –X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

[0085] With respect to the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any group or selected embodiment described above, in some embodiments, R 1 is selected from the group consisting of phenyl, pyridyl, pyrimidinyl and pyrazinyl; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

[0086] With respect to the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any of the above embodiments, in some further embodiments, the ring vertices a and b are CH; R 2b is H; the ring vertex e is C(R 2e ), and R 2a and R 2e Independently selected from the following group: C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, -OC 1-6 Halogenated alkyl, -SC 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6 Alkyl, -C 1-6 Alkyl-SC 1-6 Alkyl, CN and halogen.

[0087] With respect to the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any of the above embodiments, in some further embodiments, the ring vertices a and b are CH; R 2b is H; the ring vertex e is C(R 2e ), and R 2a and R 2e Independently selected from the following group: C 1-6 Alkyl, C 1-6 Alkoxy and halogen.

[0088] With respect to the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any of the above embodiments, in some further embodiments, subscript n is 0, 1 or 2, and if R is present 5 , each R 5 Selected from the group consisting of F, Cl, CN, C 1-4 Alkyl and C 1-4 In another further embodiment, the subscript n is 0, 1 or 2, and if R is present 5 , each R 5 Selected from the group consisting of F, Cl, CN, CH3 and OCH3.

[0089] With respect to the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any of the above embodiments, in some further embodiments, the subscript m is 0, 1 or 2, and if R is present 3 , each R 3 C 1-4 alkyl.

[0090] In a particular group of embodiments of the compounds of formula (I), or pharmaceutically acceptable salts thereof, R 1 is selected from the group consisting of phenyl or pyridine, wherein the group -X 1 -R1 Optionally 1 to 4 R x Substituted by a substituent; the ring vertices a and b are CH; R 2b is H; the ring vertex e is C(R 2e ), and R 2a and R 2e Independently selected from the following group: C 1-6 Alkyl, C 1-6 alkoxy and halogen; m is 0, 1 or 2, and if present R 3 , each R 3 CH3; R 4 Select from the following group:

[0091]

[0092] n is 0, 1 or 2, and if R is present 5 , each R 5 Selected from the group consisting of F, Cl, CN, CH3 and OCH3.

[0093] In some embodiments of the compound of formula (I), or a pharmaceutically acceptable salt thereof, R 1 Select from the following group:

[0094]

[0095] In some embodiments of the compound of formula (I), or a pharmaceutically acceptable salt thereof, -X 1 -R 1 Select from the following group:

[0096]

[0097] In some embodiments of the compound of formula (I), or a pharmaceutically acceptable salt thereof, R 1 Select from the following group:

[0098]

[0099] In some embodiments of the compound of formula (I), or a pharmaceutically acceptable salt thereof, R 1 Select from the following group:

[0100]

[0101] In some embodiments of the compound of formula (I), or a pharmaceutically acceptable salt thereof, R 1 Select from the following group:

[0102]

[0103] With respect to the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any of the above embodiments, in some further embodiments, the group Select from the following group:

[0104]

[0105] Regarding the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any of the above embodiments, in some further embodiments, wherein n is 0.

[0106] With respect to the compound of formula (I), or a pharmaceutically acceptable salt thereof, and any of the above embodiments, in some further embodiments, the subscript n is 2 and the two R on the same carbon atom are 3 Combine to form an oxo group (=O).

[0107] In some selected embodiments, the compound provided herein, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:

[0108]

[0109] In some embodiments, the compound of Formula (I) is a compound described in the Examples section and the accompanying Tables.

[0110] Preparation of compounds

[0111] Certain compounds of the present invention can be prepared according to the methods described in the Examples section herein. In addition, the synthesis of certain intermediate compounds useful in preparing compounds of the present invention is also described.

[0112] B. Pharmaceutical Compositions

[0113] In addition to the above-mentioned compounds, compositions for modulating C5a activity in humans and animals generally contain a pharmaceutical carrier or diluent.

[0114] As used herein, the term "composition" refers to a product comprising specified ingredients in specified amounts, as well as any product based directly or indirectly on the manufacture of a combination of specified ingredients in specified amounts. "Pharmaceutically acceptable" means that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0115] The pharmaceutical composition for administering the compounds of the invention can be conveniently present in unit dosage form and can be prepared by any method well known in the pharmaceutical and administration fields. All methods include the step of combining the active ingredient with a carrier constituting one or more auxiliary components. In general, the pharmaceutical composition is prepared by uniformly and closely combining the active ingredient with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, the product is made into a desired formulation. In the pharmaceutical composition, the amount of the active target compound is sufficient to produce a desired effect on the course or condition of the disease.

[0116] The pharmaceutical composition containing the active ingredient can be in a form suitable for oral use, for example, as tablets, lozenges, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions and self-emulsifiers (as described in U.S. Patent Application 2002-0012680), hard or soft capsules, syrups, elixirs, solutions, oral patches, oral gels, chewing gums, chewable tablets, effervescent powders and effervescent tablets. Compositions for oral use can be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions can include one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, antioxidants and preservatives to provide pharmaceutically elegant and palatable preparations. Tablets contain active ingredients mixed with nontoxic pharmaceutically acceptable excipients suitable for making tablets. These excipients can be, for example, inert diluents such as cellulose, silicon dioxide, aluminum oxide, calcium carbonate, sodium carbonate, glucose, mannitol, sorbitol, lactose, calcium phosphate or sodium phosphate; granulating agents and disintegrants, for example, corn starch, or alginic acid; binding agents, such as PVP, cellulose, PEG, starch, gelatin or gum arabic, and lubricants, such as magnesium stearate, stearic acid or talc. Tablets can be uncoated or can be coated, enteric or otherwise, by known techniques to delay disintegration and absorption in the gastrointestinal tract to provide a longer period of sustained action. For example, time-delay materials such as glyceryl monostearate or glyceryl distearate can be used. They can also be coated by the techniques described in U.S. Pat. Nos. 4,256,108, 4,166,452 and 4,265,874 to form osmotic therapeutic tablets for controlled release.

[0117] For oral preparations, hard gelatin capsules may also be used, in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or soft gelatin capsules, in which the active ingredient is mixed with water or an oily medium such as peanut oil, liquid paraffin or olive oil. In addition, emulsions may be prepared with water-immiscible ingredients (such as oils) and stabilized with surfactants (such as monoglycerides, PEG esters, etc.).

[0118] Aqueous suspensions contain the active substance in admixture with excipients suitable for making aqueous suspensions. Such excipients are suspending agents, such as sodium carboxymethylcellulose, ethylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum arabic; dispersants or wetting agents, which may be naturally occurring phospholipids (e.g., lecithin) or condensation products of alkylene oxides with fatty acids (e.g., polyoxyethylene stearate), or condensation products of alkylene oxides with long-chain fatty alcohols (e.g., heptadecaethyleneoxycetanol) or condensation products of alkylene oxides with partial esters derived from fatty acids and hexitol (e.g., polyoxyethylene sorbitol monooleate) or condensation products of alkylene oxides with partial esters derived from fatty acids and hexitol anhydrides (e.g., polyvinyl sorbitan monooleate). Aqueous suspensions may also contain one or more preservatives, such as ethyl, or n-propyl, parabens, one or more colorants, one or more flavoring agents, and one or more sweeteners (e.g., sucrose or saccharin).

[0119] Oily suspensions can be prepared by suspending the active ingredient in a vegetable oil (e.g., peanut oil, olive oil, sesame oil, or coconut oil) or a mineral oil (e.g., liquid paraffin). The oily suspension may contain a thickening agent, such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners and flavorings such as those described above may be added to provide a formulation suitable for oral administration. These compositions may be preserved by adding an antioxidant such as ascorbic acid.

[0120] Dispersible powders and granules suitable for preparing an aqueous suspension by adding water provide the active ingredient in admixture with a dispersing or wetting agent, a suspending agent and one or more preservatives. Examples of suitable dispersing or wetting agents and suspending agents are described above. Other excipients may be present, such as sweeteners, flavoring agents and coloring agents.

[0121] Pharmaceutical composition of the present invention can also be in the form of water-in-oil emulsion.Oil phase can be vegetable oil (for example olive oil or peanut oil) or mineral oil (for example liquid paraffin) or mixture of these.Suitable emulsifier can be naturally occurring gum, for example gum arabic or tragacanth, naturally occurring phosphatide for example soybean, lecithin and fatty acid derived ester or partial ester and hexitol anhydride, for example sorbitol monooleate, the condensation product of described partial ester and ethylene oxide, for example polyoxyethylene sorbitan monooleate.Emulsion can also contain sweetener and flavoring.

[0122] Syrups and elixirs can be formulated with sweeteners (e.g., glycerol, propylene glycol, sorbitol, or sucrose). The formulations may also contain demulcents, preservatives, and flavoring and coloring agents. Oral solutions can be prepared by combining, for example, cyclodextrins, PEGs, and surfactants.

[0123] The pharmaceutical composition can be in the form of a sterile injectable aqueous solution or oily suspension. The suspension can be prepared according to known techniques with those suitable dispersants or wetting agents and the above-mentioned suspending agents for sterile injection preparations. It can also be a sterile injection solution or suspension in a non-toxic parenteral acceptable diluent or solvent, such as a solution in 1,3-butanediol. Among acceptable carriers and solvents, water, Ringer's solution and isotonic sodium chloride solution can be used. In addition, sterile, fixed oils are generally used as solvents or suspension media. For this purpose, any mild non-volatile oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids (such as oleic acid) can be used to prepare injections.

[0124] The compounds of the present invention can also be administered rectally in the form of suppositories. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient, which is solid at room temperature but liquid at rectal temperature, and therefore will melt in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycol. In addition, the compound can be administered through the eye via a solution or ointment. Transdermal administration of the compounds of the present invention can also be achieved by means of iontophoresis patches, etc. For topical use, creams, ointments, gels, solutions or suspensions containing the compounds of the present invention, etc., can be used. As used herein, topical application also refers to the use of mouthwashes and gargles.

[0125] The compound of the present invention can also be coupled with a suitable carrier, and the carrier is a suitable polymer as a targeted drug carrier. The polymer can include polyvinyl pyrrolidone, pyran copolymer, polyhydroxypropyl methacrylamide-phenol, polyhydroxyethyl-asparagine-phenol or the polyethylene oxide-polylysine substituted with palmitoyl residues. In addition, the compound of the present invention can be coupled to a carrier, and the carrier is a class of biodegradable polymers that can be used to realize the controlled release of medicine, for example, polylactic acid, polyglycolic acid, polylactic acid and polyglycolic acid copolymers, polycaprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked or amphiphilic block copolymers of hydrogel. Polymer and semipermeable polymer matrix can form shaped articles, such as valve, stent, pipeline, prosthesis, etc. In one embodiment of the invention, the compounds of the present invention are coupled to form stent or coated stent device polymer or semipermeable polymer matrix.

[0126] The pharmaceutical composition of the present invention can be formulated with one or more additional therapeutic agents. The one or more additional therapeutic agents are selected from the group consisting of corticosteroids, steroids, immunosuppressants, immunoglobulin G agonists, dipeptidyl peptidase IV inhibitors, lymphocyte function antigen-3 receptor antagonists, interleukin-2 ligands, interleukin-1β ligand inhibitors, IL-2 receptor α subunit inhibitors, HGF gene stimulators, IL-6 antagonists, IL-5 antagonists, α1-antitrypsin stimulators, cannabinoid receptor antagonists, histone deacetylase inhibitors, AKT protein kinase inhibitors, CD20 inhibitors, Abl tyrosine kinase inhibitors, JAK tyrosine kinase inhibitors, TNFα ligand inhibitors, heme modulators, TNF antagonists, proteasome inhibitors, CD 3 regulators, Hsp70 family inhibitors, immunoglobulin agonists, CD30 antagonists, tubulin antagonists, sphingosine-1-phosphate receptor-1 agonists, connective tissue growth factor ligand inhibitors, caspase inhibitors, adrenocorticotropic hormone ligands, Btk tyrosine kinase inhibitors, complement C1s subcomponent inhibitors, erythropoietin receptor agonists, B lymphocyte stimulating ligand inhibitors, cyclin-dependent kinase-2 inhibitors, P-selectin glycoprotein ligand-1 stimulators, mTOR inhibitors, elongation factor factor)2 inhibitors, cell adhesion molecule inhibitors, factor XIII agonists, calcineurin inhibitors, immunoglobulin G1 agonists, inosine monophosphate dehydrogenase inhibitors, complement C1s subcomponent inhibitors, thymidine kinase modulators, cytotoxic T-lymphocyte protein-4 modulators, angiotensin II receptor antagonists, angiotensin II receptor modulators, TNF superfamily receptor 12A antagonists, CD52 antagonists, adenosine deaminase inhibitors, T-cell differentiation antigen CD6 inhibitors, FGF-7 ligands, dihydroorotate dehydrogenase inhibitors, Syk tyrosine kinase inhibitors, interferon type I receptor antagonists, interferon α ligand inhibitors, macrophage migration inhibitory factorfactor inhibitors, integrin α-V / β-6 antagonists, cysteine ​​protease stimulators, p38MAP kinase inhibitors, TP53 gene inhibitors, Shiga-like toxin I inhibitors, fucosyltransferase 6 stimulators, interleukin 22 ligands, IRS1 gene inhibitors, protein kinase C stimulators, protein kinase Cα inhibitors, CD74 antagonists, immunoglobulin gamma Fc receptor IIB antagonists, T-cell antigen CD7 inhibitors, CD95 antagonists, N-acetylmannosamine kinase stimulators, cardiotrophin-1 ligands, leukocyte elastase elastase) inhibitors, CD40 ligand receptor antagonists, CD40 ligand modulators, IL-17 antagonists, TLR-2 antagonists, mannan-binding lectin serine protease-2 (MASP-2) inhibitors, factor B inhibitors, factor D inhibitors, C3aR modulators, C5aR2 modulators, T cell receptor antagonists, PD-1 inhibitors, PD-L1 inhibitors, TIGIT inhibitors, TIM-3 inhibitors, LAG-3 inhibitors, VISTA inhibitors, STING agonists, IDO inhibitors, adenosine receptor modulators, CD39 inhibitors, CD73 inhibitors, chemokine receptor antagonists (especially CXCR1, CXCR2, CXCR3, CXCR4, CXCR7, CCR1, CCR2, CCR3, CCR4, CCR5, CCR7, CCR7, CCR9, CX3CR1 and CXCR6), and combinations thereof.

[0127] In some embodiments, the one or more additional therapeutic agents are selected from the group consisting of obinutuzumab, rituximab, ocrelizumab, cyclophosphamide, prednisone, hydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortol pivalate, prednisolone, methylprednisolone, triamcinolone acetonide, triamcinolone acetonide, cyclophosphamide, cyclophosphamide, prednisone, hydrocortisone acetate, hydrocortisone acetate, tixocortol pivalate, prednisolone, methylprednisolone, triamcinolone acetonide ... alcohol), mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinoloneacetonide, halcinonide, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortolone, hydrocortisone-17-valerate, halometasone, alclometasone dipropionate, beclomethasone, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, clobetasol 17-propionate, fluocortolone hexanoate, fluocortolone pivalate, fluprednidene acetate acetate), hydrocortisone-17-butyrate, hydrocortisone-17-aceponate, hydrocortisone-17-buteprate, ciclesonide and prednicarbate, GB-0998, immuglo, begelomab, alefacept, aldesleukin, gevokizumab, daclizumab, basiliximab, inolimomab, beperminogen activator, perplasmid), sirukumab, tocilizumab, clazakizumab, mepolizumab, fingolimod, panobinostat, triciribine, nilotinib, imatinib, tofacitinib, momelotinib,peficitinib, itacitinib, infliximab, PEG-bHb-CO, etanercept, ixazomib, bortezomib, muromonab, otelixizumab, gusperinmus, brentuximab vedotin), Ponesimod, KRP-203, FG-3019, emricasan, ACTH, ibrutinib, cinryze, conestat, methoxypolyethylene glycol-epoetin beta, belimumab, blisibimod, atacicept, seliciclib, neihulizumab, everolimus, sirolimus, denileukin diftitox), LMB-2, natalizumab, catridecacog, cyclosporine, tacrolimus, voclosporin, vaclosporin, canakinumab, mycophenolate, mizoribine, CE-1145, TK-DLI, abatacept, belatacept, olmesartan medoxomil, sparsentan, TXA-127, BIIB-023, alemtuzumab, pentostatin, itolizumab, palifermin, leflunomide, PRO-140, cenicriviroc, fostamatinib, anirumab frolumab), sifalimumab, BAX-069, BG-00011, losmapimod, QPI-1002, ShigamAb, TZ-101, F-652, reparixin, ladarixin, PTX-9908, aganirsen, APH-703, sotrastaurin, sotrastaurin, milatuzumab, SM-101, T-Guard, APG-101, DEX-M74, cardiotrophin-1, tiprelestat, ASKP-1240, BMS-986004,HPH-116, KD-025, OPN-305, TOL-101, defibrotide, pomalidomide, thymic globulin, laquinimod, remestemcel-L, equine antithymocyte immunoglobulin, Stempeucel, LIV-γ, Octagam 10%, t2c-001, 99mTc-sestamibi (sestamibi), Clairyg, Prosorba, pomalidomide, laquinimod, teplizumab, FCRx, solnatide, foralumab, ATIR-101, BPX-501, ACP-01, ALLO-ASC-DFU, irbesartan + propagerm anium), ApoCell, cannabidiol, RGI-2001, orotic acid (saratin), anti-CD3 bivalent antibody-diphtheria toxin conjugate, NOX-100, LT-1951, OMS721, ALN-CC5, ACH-4471, AMY-101, Acthar gel, and CD4+CD25+ regulatory T cells, MEDI7814, P32, P59, pembrolizumab, nivolumab ), atezolizumab, avelumab, durvalumab, CCX354, CCX721, CCX9588, CCX140, CCX872, CCX598, CCX6239, CCX587, CCX624, CCX282, CCX025, CCX507, CCX430, CCX765, CCX758, CCX771, CCX662, CCX650, and combinations thereof. Further discussion of combination therapy is included in the "Methods of Use" section of this application. ,

[0128] A. How to use

[0129] The compounds of the present invention can be used as agonists, (preferably) antagonists, partial agonists, inverse agonists of C5a receptors in various situations in vivo and in vitro. In one embodiment, the compound of the present invention is a C5aR antagonist, which can be used to inhibit the binding of C5a receptor ligands (such as C5a) to C5a receptors in vitro or in vivo. In general, the method includes contacting the C5a receptor with an effective amount of one or more C5a receptor modulators provided by the present invention, when the C5a receptor ligand is present in an aqueous solution and other conditions suitable for the binding of the ligand to the C5a receptor. The C5a receptor can be present in a suspension (e.g., in an isolated membrane or cell preparation), in a cultured or isolated cell, or in a tissue or organ.

[0130] Preferably, the amount of C5a receptor modulator contacted with the receptor should be sufficient to inhibit the binding of C5a to the C5a receptor in vitro as determined (eg, using a radioligand binding assay, calcium mobilization assay, or chemotaxis assay described herein).

[0131] In one embodiment of the invention, the C5a modulators of the invention can be used to modulate (preferably inhibit) the signal transduction activity of the C5a receptor, for example, by contacting one or more compounds of the invention with the C5a receptor (in vitro or in vivo) under conditions suitable for binding of the modulator to the receptor. The receptor can be present in solution or suspension, in a cultured or isolated cell preparation, or in a patient's body. Any modulation of signal transduction activity can be assessed by measuring the effect on calcium ion calcium mobilization or by measuring the effect on C5a receptor-mediated cellular chemotaxis. In general, an effective amount of a C5a modulator is an amount sufficient to modulate C5a receptor signal transduction activity in vitro in a calcium mobilization assay or C5a receptor-mediated cellular chemotaxis in a migration assay.

[0132] When the compounds of the present invention are used to inhibit C5a receptor-mediated cellular chemotaxis, preferably chemotaxis of leukocytes (such as neutrophils), in an in vitro chemotaxis assay, the method comprises contacting one or more compounds of the present invention with leukocytes (especially primate leukocytes, especially human leukocytes). Preferably, the concentration is sufficient to inhibit the chemotaxis of leukocytes in an in vitro chemotaxis assay, so that, as described above, the level of chemotaxis observed in the control assay is significantly higher than the level of chemotaxis observed in the assay with the addition of the compounds of the present invention.

[0133] In another embodiment, the compounds of the present invention are further used to treat patients with diseases that respond to C5a receptor modulation. As used herein, the terms "treatment" or "therapy" include disease-modifying treatment and symptomatic treatment, any of which can be preventive (i.e., before the onset of symptoms, in order to prevent, delay or reduce the severity of symptoms) or therapeutic (i.e., after the onset of symptoms, in order to reduce the severity and / or duration of symptoms). As used herein, if the modulation of C5a receptor activity results in a reduction in the inappropriate activity of the C5a receptor, it is considered to be a disease that "responds to C5a receptor modulation." As used herein, the term "patient" includes primates (particularly humans), domestic companion animals (such as dogs, cats, horses, etc.) and livestock (such as cattle, pigs, sheep, etc.) at the doses described herein.

[0134] Conditions that can be treated by C5a modulation:

[0135] Autoimmune disorders - For example, rheumatoid arthritis, systemic lupus erythematosus, Guillain-Barré syndrome, pancreatitis, lupus nephritis, lupus glomerulonephritis, psoriasis, Crohn's disease, vasculitis, irritable bowel syndrome, dermatomyositis, multiple sclerosis, bronchial asthma, dense deposit disease, pemphigus, pemphigoid, scleroderma, myasthenia gravis, autoimmune hemolytic and thrombocytopenic states, Goodpasture's syndrome (and associated glomerulonephritis and pulmonary hemorrhage), C3-glomerulopathy, C3-glomerulonephritis, membranoproliferative glomerulonephritis, Kawasaki disease, IGs nephropathy, immune vasculitis, tissue transplant rejection, graft-versus-host disease, hyperacute rejection of transplanted organs, etc.

[0136] Inflammatory diseases and related conditions- For example, neutropenia, sepsis, septic shock, Alzheimer's disease, multiple sclerosis, neutrophilia, stroke, inflammatory bowel disease (IBD), inflammation associated with severe burns, lung injury, and ischemia-reperfusion injury, osteoarthritis, as well as acute (adult) respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), systemic inflammatory response syndrome (SIRS), atopic dermatitis, psoriasis, chronic urticaria and multiple organ dysfunction syndrome (MODS), hemolytic uremic syndrome, atypical hemolytic uremic syndrome (aHUS). Also included are pathological sequelae associated with insulin-dependent diabetes (including diabetic retinopathy), lupus nephropathy, Heyman nephritis, membranous nephritis and other forms of glomerulonephritis, contact sensitivity reactions, and inflammation caused by contact of blood with artificial surfaces caused by complement activation, for example, during extracorporeal blood circulation (e.g., during hemodialysis or through a heart-lung machine, for example, associated with vascular surgery such as coronary artery bypass grafting or heart valve replacement), or with other artificial vessels or container surfaces (e.g., ventricular assist devices, artificial heart machines, infusion tubes, blood bags, plasma exchange, platelet collection, etc.). Also included are diseases associated with ischemia / reperfusion injury, such as diseases caused by transplants (including solid organ transplants) and syndromes such as ischemic reperfusion injury / ischemic colitis and cardiac ischemia. The compounds of the present invention can also be used to treat age-related macular degeneration (Hageman et al., PNAS 102: 7227-7232, 2005).

[0137] Cardiovascular and cerebrovascular diseases - For example, myocardial infarction, coronary thrombosis, vascular occlusion, postoperative vascular reocclusion, atherosclerosis, traumatic central nervous system injury and ischemic heart disease. In one embodiment, an effective amount of the compounds of the present invention can be administered to patients at risk of myocardial infarction or thrombosis (i.e., patients with one or more identified risk factors for myocardial infarction or thrombosis, such as, but not limited to, obesity, smoking, hypertension, hypercholesterolemia, a previous or genetic history of myocardial infarction or thrombosis) to reduce the risk of myocardial infarction or thrombosis.

[0138] Neoplastic diseases or conditions- For example, melanoma, lung cancer, lymphoma, sarcoma, carcinoma, fibrosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, angiosarcoma, lymphangiosarcoma, synovioma, mesothelioma, meningioma, leukemia, lymphoma, leiomyosarcoma, rhabdomyosarcoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, papillary carcinoma, cystadenocarcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatocellular carcinoma, transitional cell carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, pleomorphic adenoma, liver cell papilloma, tubular adenoma, cystadenoma, papilloma, adenoma, leiomyoma, rhabdomyoma, hemangioma, lymphangioma, osteoma, chondroma, lipoma and fibroma.

[0139] Vasculitis -Vasculitis disease characterized by vascular inflammation. The infiltration of leukocytes causes the destruction of vascular walls, and the complement pathway is considered to play a major role in the damage caused by starting leukocyte migration and showing at the site of inflammation (Vasculitis, second edition, Ball and Bridges edited, Oxford University Press, 47-53 pages, 2008). Compound provided by the invention can be used to treat leukocytic vasculitis, antineutrophil cytoplasmic antibodies (ANCA)-related vasculitis, immune vasculitis Wegener's granulomatosis, microscopic polyangiitis, allergic granulomatous (Churg-Strausssyndrome), Henoch-Schonlein purpura (Henoch-Schonlein purpura), polyarteritis nodosa (polyateritisnodosa), rapidly progressive glomerulonephritis (RPGN), cryoglobulinemia, giant cell arteritis (GCA), Behcet's disease and Tak's arteritis (TAK).

[0140] HIV infection and AIDS --The C5a receptor modulator provided by the present invention can be used to inhibit HIV infection, delay the progression of AIDS or reduce the symptoms or severity of HIV infection and AIDS.

[0141] Neurodegenerative and related diseases --On the other hand, the C5a antagonists provided by the present invention can be used to treat Alzheimer's disease, multiple sclerosis, and cognitive decline associated with cardiopulmonary bypass surgery and related procedures.

[0142] In one embodiment of the invention, the compounds of the invention may be used to treat a disease selected from the group consisting of sepsis (and related conditions), COPD, rheumatoid arthritis, lupus nephritis and multiple sclerosis.

[0143] The therapeutic methods provided by the present invention generally include administering to a patient an effective amount of one or more compounds provided by the present invention. Suitable patients include those patients suffering from or susceptible to (i.e., preventive treatment) the symptoms or diseases determined herein. Typical patients for treatment as described herein include mammals, particularly primates, and especially humans. Other suitable patients include domestic companion animals, for example, dogs, cats, horses, etc., or livestock such as cattle, pigs, sheep, etc.

[0144] In general, the treatment methods provided herein include administering to a patient an effective amount of one or more compounds provided herein. In a preferred embodiment, the compounds of the present invention are preferably administered orally or topically to a patient (e.g., a human). The effective amount may be an amount sufficient to modulate the activity of the C5a receptor and / or an amount sufficient to reduce or alleviate the symptoms presented by the patient. Preferably, the amount administered is sufficient to produce a sufficiently high plasma concentration of the compound (or its active metabolite, if the compound is a prodrug) to detectably inhibit chemotaxis of leukocytes (such as neutrophils) in vitro. The treatment regimen may vary depending on the compound used and the specific condition to be treated; for the treatment of most conditions, the preferred daily administration frequency is 4 times or less. In general, a twice-daily dosage regimen is more preferred, and once-daily administration is particularly preferred. However, it should be understood that the specific dosage level and treatment regimen for any particular patient will depend on various factors, including the activity of the specific compound used, age, weight, general health, sex, diet, route of administration, excretion rate, drug combination (i.e., other drugs given to the patient) and the severity of the specific disease being treated, as well as the judgment of the prescribing physician. In general, it is preferred to use the minimum dose sufficient to provide effective treatment. Patients can generally be monitored for effectiveness of treatment using medical or veterinary criteria appropriate to the condition being treated or prevented.

[0145] Dosage levels of about 0.1 mg to about 140 mg per kilogram of body weight per day are useful for treating or preventing diseases involving pathogenic C5a activity (about 0.5 mg to about 7 g per person per day). The amount of active ingredient that may be combined with a carrier material to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. Unit dosage forms typically contain between about 1 mg and about 500 mg of the active ingredient. For oral, transdermal, intravenous or subcutaneous administration of the compound, preferably, sufficient compound is administered to achieve a serum concentration of 5 ng (nanograms) / mL-10 μg (micrograms) / mL serum, more preferably, sufficient compound should be administered to achieve a serum concentration of 20 ng-1 μg / ml serum, and most preferably, sufficient compound should be administered to achieve a serum concentration of 50 ng / ml-200 ng / ml serum. For direct injection into the synovium (for the treatment of arthritis) sufficient compound should be administered to achieve a local concentration of about 1 micromolar.

[0146] The frequency of dosage can also vary depending on the compound used and the specific disease being treated. However, for the treatment of most conditions, a dosage regimen of 4 times a day, 3 times a day, or preferably less, once a day or twice a day is particularly preferred. However, it should be understood that the specific dosage level for any particular patient will depend on various factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, administration time, administration route, and excretion rate, the severity of the specific disease being treated by the drug combination (i.e., other drugs given to the patient), and other factors including the judgment of the prescribing physician.

[0147] Combination therapy

[0148] The compounds disclosed in the present invention can be used in combination with one or more other therapeutic agents for treating, preventing, suppressing or improving the diseases or conditions available for the compounds and compositions of the present invention. Such one or more other therapeutic agents can be administered by the compounds or compositions of the present invention simultaneously or successively with their commonly used approaches and amounts. When the compounds or compositions of the present invention are used simultaneously with one or more other drugs, it is preferred that the pharmaceutical composition comprises such other drugs except the compounds or compositions of the present invention. Therefore, the pharmaceutical composition of the present invention includes those that also comprise one or more other active ingredients or therapeutic agents except the compounds or compositions of the present invention.

[0149] Examples of the one or more additional therapeutic agents are: corticosteroids, steroids, immunosuppressants, immunoglobulin G agonists, dipeptidyl peptidase IV inhibitors, lymphocyte function antigen-3 receptor antagonists, interleukin-2 ligands, interleukin-1β ligand inhibitors, IL-2 receptor α subunit inhibitors, HGF gene stimulators, IL-6 antagonists, IL-5 antagonists, α1-antitrypsin stimulators, cannabinoid receptor antagonists, histone deacetylase inhibitors, AKT protein kinase inhibitors, CD20 inhibitors, Abl tyrosine kinase inhibitors, JAK tyrosine kinase inhibitors, TNFα ligand inhibitors, heme modulators, TNF antagonists, proteasome inhibitors, CD3 modulators, Hsp70 family family inhibitors, immunoglobulin agonists, CD30 antagonists, tubulin antagonists, sphingosine 1-phosphate receptor-1 agonists, connective tissue growth factor ligand inhibitors, caspase inhibitors, adrenocorticotropic hormone ligands, Btk tyrosine kinase inhibitors, complement C1s subcomponent inhibitors, erythropoietin receptor agonists, B lymphocyte stimulating ligand inhibitors, cyclin-dependent kinase-2 inhibitors, P-selectin glycoprotein ligand-1 stimulators, mTOR inhibitors, elongation factor 2 inhibitors, cell adhesion molecule inhibitors, factor XIII agonists, calcineurin inhibitors, immunoglobulin G1 agonists, inosine monophosphate dehydrogenase inhibitors, complement C1s subcomponent inhibitors, thymidine kinase modulators , cytotoxic T-lymphocyte protein-4 modulators, angiotensin II receptor antagonists, angiotensin II receptor modulators, TNF superfamily receptor 12A antagonists, CD52 antagonists, adenosine deaminase inhibitors, T-cell differentiation antigen CD6 inhibitors, FGF-7 ligands, dihydroorotate dehydrogenase inhibitors, Syk tyrosine kinase inhibitors, interferon type I receptor antagonists, interferon α ligand inhibitors, macrophage migration inhibitory factor inhibitors, integrin α-V / β-6 antagonists, cysteine ​​protease stimulators, p38MAP kinase inhibitors, TP53 gene inhibitors, Shiga-like toxin I inhibitors, fucosyltransferase 6 stimulators, interleukin 22 ligands, IRS1 gene inhibitors, protein kinases C stimulator, protein kinase Cα inhibitor, CD74 antagonist, immunoglobulin gamma Fc receptor IIB antagonist, T-cell antigen CD7 inhibitor, CD95 antagonist, N-acetyl mannosamine kinase stimulator, cardiotrophin-1 ligand, leukocyte elastase inhibitor, CD40 ligand receptor antagonist, CD40 ligand modulator, IL-17 antagonist, TLR-2 antagonist, mannan-binding lectin serine protease-2 (MASP-2) inhibitor, factor B inhibitor, factor D inhibitor, C3aR modulator, C5aR2 modulator, T cell receptor antagonist, PD-1 inhibitor, PD-L1 inhibitor, TIGIT inhibitor, TIM-3 inhibitor, LAG-3 inhibitor,VISTA inhibitors, STING agonists, IDO inhibitors, adenosine receptor modulators, CD39 inhibitors, CD73 inhibitors, chemokine receptor antagonists (especially CXCR1, CXCR2, CXCR3, CXCR4, CXCR7, CCR1, CCR2, CCR3, CCR4, CCR5, CCR7, CCR7, CCR9, CX3CR1 and CXCR6), and combinations thereof.

[0150] In some embodiments, the additional therapeutic agent used in the methods of treatment described herein is selected from the group consisting of obinutuzumab, rituximab, ocrelizumab, cyclophosphamide, prednisone, hydrocortisone, hydrocortisone acetate, cortisone acetate, sulhydrocortisone pivalate, prednisolone, methylprednisolone, triamcinolone acetonide, triamcinolone, mometasone, amcinonide, budesonide, desonide, fluocinolone acetonide, fluocinolone acetonide, halcinonide, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortolone, hydrocortisone-17-valerate, halometasone, alclomethasone dipropionate, beclomethasone, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, Clobetasol 17-propionate, fluocortolone hexanoate, fluocortolone trimethylacetate, fluprednidene acetate, hydrocortisone-17-butyrate, hydrocortisone-17-acetate, hydrocortisone-17-propionate, ciclesonide and prednicarbate, GB-0998, imeg antibody, begromab, alecept, aldesleukin, gvarizumab, daclizumab, basiliximab, inomumab, bepluminogen plasmid, serucumab, tocilizumab, clazazumab, mepolizumab, fingolimod, panobinostat, triciribine, nilotinib, imatinib, tofacitinib, mometinib, pegol, itatinib, infliximab, PEG-bHb-CO, etanercept, ixazomib, bortezomib, moro monoclonal antibody, oriximab, gustation, bentoximab, prednismol, KRP-203, FG-3019, enlicasin, adrenocorticotropic hormone, ibrutinib, sinluze, conastat, methoxypolyethylene glycol-epoetin beta, belimumab, bilismol, atacicept, celecoxib, nehulizumab, everolimus, sirolimus, denileukin, LMB-2, natalizumab, cadrecol, cyclosporine, tacrolimus, waclosporin, waclosporin, canakinumab, mycophenolate mofetil, mizoribine, CE-1145, TK-DLI, abatacept, belatacept, olmesartan medoxomil, spartantan, TXA-127, BIIB-023, alemtuzumab, pentostatin, itulizumab, par Rifumin, leflunomide, PRO-140, senecalivarox, flustatinib, aniluomab, sefavirenz, BAX-069, BG-00011, rosmapimod, QPI-1002, shigamonab, TZ-101, F-652, rapamycin, radarexin, PTX-9908, aganelson, APH-703, sotastatin, sotastatin, milatuzumab, SM-101, T-Guard, APG-101, DEX-M74, cardiotrophin-1, tepistatin, ASKP-1240, BMS-986004, HPH-116, KD-025, OPN-305, TOL-101, defibrotide, pomalidomide,Thymoglobulin, laquinimod, ramestecil-L, equine antithymocyte immunoglobulin, streptocil, LIV-γ, octagon 10%, t2c-001, 99mTc-cestamid, cleyig, proloxobar, pomalidomide, laquinimod, teplizumab, FCRx, sonatinide, vlalimumab, ATIR-101, BPX-501, ACP-01, ALLO-ASC-DFU, irbesartan + propagandam, ApoCell, cannabidiol, RGI-2001, orotic acid, anti-CD3 bivalent antibody-diphtheria toxin conjugate, NOX-100, LT-1951, OMS72 1. ALN-CC5, ACH-4471, AMY-101, Aksar gel, and CD4+CD25+ regulatory T cells, MEDI7814, P32, P59, pembrolizumab, nivolumab, atezolizumab, avelumab, durvalumab, CCX354, CCX721, CCX9588, CCX140, CCX872, CCX598, CCX6239, CCX587, CCX624, CCX282, CCX025, CCX507, CCX430, CCX765, CCX758, CCX771, CCX662, CCX650, and combinations thereof.

[0151] The disease or condition being treated will determine the other therapeutic agent or agents most suited for administration in combination with the compounds of the invention - such determinations can be made by one skilled in the art.

[0152] The weight ratio of the compound of the present invention to the second active ingredient can vary and depends on the effective dose of each ingredient. In general, the respective effective doses are used. Thus, for example, when the compound of the present invention is combined with an NSAID, the weight ratio of the compound of the present invention to the NSAID is generally from about 1000:1 to about 1:1000, preferably, from about 200:1 to about 1:200. The combination of the compound of the present invention and other active ingredients is generally also within the above range, but in each case, the effective dose of each active ingredient should be used.

[0153] Non-medical applications

[0154] In another aspect of the invention, the compounds of the invention can be used in a variety of non-pharmaceutical in vitro and in vivo applications. For example, the compounds of the invention can be labeled and used as probes for detecting and localizing C5a receptors (cell preparations or tissue section samples). The compounds of the invention can also be used as positive controls for C5a receptor activity assays, i.e., as standards for determining the ability of candidate agents to bind to C5a receptors, or as radioactive tracers for positron emission tomography (PET) imaging or single photon emission computed tomography (SPECT). The methods can be used to characterize C5a receptors in vivo. For example, C5a receptor modulators can be labeled using any of a variety of well-known techniques (e.g., radiolabeling with a radionuclide such as tritium), and the samples can be incubated for a suitable incubation time (e.g., determined by the time course of the first determination of binding). After incubation, unbound compounds are removed (e.g., by washing), and bound compounds are detected using any method suitable for the label used (e.g., autoradiography or scintillation counting of radiolabeled compounds; luminescent and fluorescent groups can be detected using spectroscopic methods). As a control, a matched sample containing the labeled compound and a greater (e.g., 10-fold greater) amount of unlabeled compound can be treated in the same manner. A greater amount of detectable label remaining in the test sample than in the control sample indicates the presence of C5a receptor in the sample. Detection assays, including receptor autoradiography (receptor mapping) of C5a receptors in cultured cells or tissue samples can be performed as described by Kuhar in Current Protocols in Pharmacology (1998) John Wiley & Sons New York, Sections 8.1.1 to 8.1.9.

[0155] The compounds provided herein can also be used in various well-known cell separation methods. For example, the modulator can be connected to the inner surface of a tissue culture plate or other support and used as an affinity ligand for in vitro fixation and separation of C5a receptors (e.g., separation of receptor-expressing cells). In a preferred application, the modulator is connected to a fluorescent marker (e.g., fluorescein), and the cells are contacted and then analyzed (or separated) by fluorescence activated cell sorting (FACS).

[0156] I. Embodiment

[0157] The following examples are intended to illustrate but not to limit the claimed invention.

[0158] The reagents and solvents used below can be obtained from commercial sources, such as Aldrich Chemical Company (Milwaukee, WI, USA). 1H-NMR spectra were recorded with a Varian Mercury 400MHz NMR spectrometer. Valid peaks are provided relative to TMS and are tabulated in the following order: multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet) and number of protons. Mass spectrometry results are reported as mass-to-charge ratios, relative abundances of each ion (in brackets). In the examples, a single m / e value reports the M+H (or labeled MH) ion containing the most common atomic isotope. In all cases, the isotope distribution (Isotope patterns) conforms to the expected formula. Electrospray ionization (ESI) mass spectrometry was performed with a Hewlett-Packard MSD electrospray mass spectrometer, with HP1100 HPLC sample delivery. The analyte is usually dissolved in methanol at 0.1 mg / mL, and 1 microliter is injected into the mass spectrometer together with the delivery solvent, and the mass spectrometer scans 100-1500 Daltons. All compounds were analyzed in positive ESI mode using acetonitrile / water containing 1% formic acid as the delivery solvent. The following compounds were also analyzed in negative ESI mode using 2 mM NH4OAc in acetonitrile / water as the delivery solvent.

[0159] The following abbreviations are used in the examples and throughout the description of the present invention:

[0160] EtOH: ethanol

[0161] EtONa: Sodium ethoxide

[0162] THF:Tetrahydrofuran

[0163] TLC: Thin layer chromatography

[0164] MeOH: Methanol

[0165] Compounds within the scope of the present invention can be synthesized as described below with various reactions known to those skilled in the art. Those skilled in the art will also recognize that alternative methods can be adopted to synthesize target compounds of the present invention, and the methods primarily described in this article are not exhaustive, but provide widely applicable and practical approaches to target compounds.

[0166] Certain molecules claimed in this patent may exist in different enantiomeric and diastereomeric forms, and all such variations of these compounds are claimed.

[0167] A detailed description of the experimental procedures used to synthesize the key compounds presented herein leads to the physical data that identified them and to the structural descriptions associated with them that illustrate the molecules.

[0168] Those skilled in the art will also recognize that acids and bases are often used during standard work-up in organic chemistry. Salts of parent compounds are sometimes produced during the experimental procedures described in this patent if they are acidic or basic in nature.

[0169] Example 1

[0170] Synthesis of 1-(4-(5-(3,5-dichloropyridin-2-yl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

[0171]

[0172] Step a: A mixture of 3-methyl-2-nitro-phenol (50.0 g, 326.5 mmol), 1-iodo-2-methyl-propane (184.0 g, 1.0 mol) and Cs2CO3 (326.0 g, 1.0 mol) in acetone (500 mL) was stirred overnight under reflux. It was then cooled to room temperature and filtered through celite. The filtrate was collected and concentrated under reduced pressure. The obtained solid was dissolved in EtOAc, washed with brine, dried over Na2SO4 and concentrated under reduced pressure to give 1-isobutoxy-3-methyl-2-nitrobenzene. 1 H NMR (400MHz, CDCl3)7.26(t,J=8.0Hz,1H),6.82(m,2H),3.78(d,J=6.8Hz,2H),2.94(s,3H),2.07(m,1H),0.98(d,J=6.4Hz,6H).

[0173] A pressure vessel containing 1-isobutoxy-3-methyl-2-nitrobenzene (130.4 g, 623.2 mmol), 10% Pd / C (25 g, 50% aqueous) and EtOH (750 mL) was stirred under 45 psi hydrogen atmosphere for 3 h. The mixture was filtered through celite. The filtrate was collected and concentrated under reduced pressure to obtain 2-isobutoxy-6-methylaniline. 11 H 18 NO[M+H] + 180.2, measured value 180.2.

[0174] WARNING: The formation of diazo compounds is potentially hazardous, please handle with caution and wear appropriate personal protective equipment!

[0175] Step b: Add isobutoxy-6-methylaniline (26.4 g, 147.3 mmol) in portions to 100 mL of concentrated hydrochloric acid at -10°C to obtain a stirrable suspension. After stirring at the same temperature for 30 min, add an aqueous solution (25 mL) of NaNO2 (12.2 g, 176.8 mmol) dropwise over 20 min to obtain the diazonium salt.

[0176] Tin(II) chloride dihydrate (83.0 g, 367.8 mmol) in concentrated HCl (120 mL) was added to the above diazonium salt in batches. The resulting mixture was then stirred at -10 °C for 10 min and then at room temperature for 1 h. The mixture was then diluted with DCM (400 mL) and water. The organic layer was separated, dried over Na2SO4, and concentrated on a rotary evaporator under reduced pressure to obtain (2-isobutoxy-6-methylphenyl)hydrazine hydrochloride. C 11 H 19 N2O[M+H] + 195.1, measured value 195.1.

[0177] Step c: To a stirred suspension of (2-isobutoxy-6-methylphenyl)hydrazine hydrochloride (8.0 g, 39.9 mmol) in EtOH (60 mL) and glacial acetic acid (12 mL, 208 mmol) at room temperature was added tert-butyl 3-cyano-4-oxopiperidine-1-carboxylate (5.0 g, 22.3 mmol). The resulting mixture was stirred at reflux for 16 h. After the solvent was removed under reduced pressure, the residue was dissolved in EtOAc and washed with aqueous NaOH (2N), brine, and dried over MgSO4. The solvent was removed under reduced pressure, and the residue was purified by silica gel flash chromatography (5 to 55% EtOAc in hexanes) to give tert-butyl 3-amino-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 22 H 33 N4O3[M+H] + 401.2, measured value 401.2.

[0178] WARNING: The formation of diazo compounds is potentially hazardous, please handle with caution and wear appropriate personal protective equipment!

[0179] Under magnetic stirring at room temperature, isoamyl nitrite (96%, 4.0 mL, 28.6 mmol) was slowly added to a mixture of tert-butyl 3-amino-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (3.0 g, 8.1 mmol), CuBr (4.0 g, 27.9 mmol) and MeCN (50 mL) in a 250 mL round-bottom flask. The resulting mixture was stirred at room temperature for 1 h, diluted with EtOAc, filtered through celite, washed with saturated aqueous NH4Cl solution, and dried over MgSO4. The solvent was removed under reduced pressure and the residue was purified by flash chromatography on silica gel (2 to 25% EtOAc in hexanes) to give tert-butyl 3-bromo-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 22 H 31 BrN3O3[M+H] + 464.1, measured value 464.2.

[0180] Step d: A mixture of 4-bromo-2,5-difluoroaniline (1.5 g, 7.2 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (2.2 g, 8.7 mmol), KOAc (1.8 g, 18.3 mmol) and Pd(dppf)Cl2 complexed with dichloromethane (580.0 mg, 0.7 mmol) in dioxane (12 mL) was stirred under nitrogen at 95° C. for 2 h. The mixture was then cooled to room temperature and filtered through celite. The filtrate was collected, concentrated under reduced pressure, and purified by silica gel flash chromatography (0 to 50% EtOAc in hexanes) to give 2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline. MS: (ES) m / z calculated value C 12 H 17 BF2NO2[M+H] + 256.1, measured value 256.2.

[0181] To a suspension of tert-butyl 3-bromo-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (125.0 mg, 0.3 mmol), 2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (75.0 mg, 0.3 mmol), Na2CO3 (85.0 mg, 0.8 mmol) in dioxane (4 mL) and water (1 mL) was added Pd(dppf)Cl2 (26.0 mg, 0.03 mmol) complexed with dichloromethane. The reaction mixture was degassed (N2) for 2 min and stirred at 95° C. under N2 for 6 h. The reaction mixture was diluted with EtOAc, filtered through celite, washed with brine, and dried over MgSO4. The solvent was removed under reduced pressure and the residue was purified by flash chromatography on silica gel (5 to 40% EtOAc in hexanes) to give tert-butyl 3-(4-amino-2,5-difluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 28 H 35 F2N4O3[M+H] + 513.3, measured value 513.3.

[0182] Step e: To a stirred solution of tert-butyl 3-(4-amino-2,5-difluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (0.5 g, 2.0 mmol) in anhydrous THF (5 mL) was added N,N-diisopropylethylamine (0.6 g, 4.8 mmol) and trimethylsilyl isocyanate (0.3 g, 2.6 mmol). The reaction mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was diluted with EtOAc, washed with brine and dried over MgSO4. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (5 to 40% EtOAc in hexanes) to give tert-butyl 3-(2,5-difluoro-4-ureidophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated C 29 H 36 F2N5O4[M+H] + 556.3, measured value 556.3.

[0183] Step f: To a solution of tert-butyl 3-(2,5-difluoro-4-ureidophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (1.2 g, 3.6 mmol) in dichloromethane (10 mL) was added 4N HCl in dioxane (3.0 mL, 12.0 mmol). The resulting mixture was stirred at room temperature for 2 h. After the reaction was complete, the solvent was diluted with water and saturated aqueous NaHCO3 and extracted with EtOAc, washed with brine and dried over MgSO4. The solvent was removed under reduced pressure to give 1-(2,5-difluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)urea. MS: (ES) m / z calculated value C 24 H 28 F2N5O2[M+H] + 456.2, measured value 456.2.

[0184] Step g: To a suspension of 1-(2,5-difluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)urea (3.0 g, 6.6 mmol) in DMSO (10 mL) was added 3,5-dichloro-5-fluoropyridine (1.6 g, 9.9 mmol) and Li2CO3 (1.9 g, 25.7 mmol) at room temperature. The resulting mixture was stirred at 90 °C for 4 h. After the reaction was complete, the mixture was cooled to room temperature and diluted with EtOAc. The organic layer was washed with brine and dried over MgSO4. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (10 to 60% EtOAc in hexanes) to give l-(4-(3,5-dichloropyridin-2-yl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea. 1H NMR (400MHz, CD3OD): δ8.15(d,J=2.3Hz,1H),8.02(dd,J=7.1,12.5Hz,1H),7.84(d,J=2.3H z,1H),7.32(t,J=7.8Hz,1H),6.92(d,J=8.2Hz,1H),6.89(d,J=7.5Hz,1H),6.82(dd,J=6.3, 11.3Hz,1H),4.87(br,2H),4.39(s,2H),3.70-3.82(m,4H),3.65(dd,J=7.1,9.0Hz,1H),3.07(t,J=6.7Hz,2H),2.02(s,3H),1.85–1.95(m,1H),0.85(d,J=7.0Hz,6H).MS:(ES) m / z calculated value C 29 H 29 Cl2F2N6O2[M+H] + 601.2, measured value 601.5.

[0185] Example 2

[0186] Synthesis of 4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzamide

[0187]

[0188] Step a: A mixture of tert-butyl 3-bromo-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (120.0 mg, 0.26 mmol), (4-cyanophenyl)boronic acid (100.0 mg, 0.68 mmol), Pd(PPh3)4 (45.0 mg, 0.04 mmol) and K2CO3 (125.0 mg, 0.9 mmol) in toluene (2 mL) and water (0.3 mL) was stirred at 110 ° C for 3 h in N2. The mixture was cooled to room temperature, quenched with water, and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 40% EtOAc in hexanes) to afford tert-butyl 3-(4-cyanophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 29 H 35 N4O3[M+H] +487.3, measured value 487.2.

[0189] Step b: A mixture of tert-butyl 3-(4-cyanophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (92.0 mg, 0.2 mmol) and a solution of 4N HCl in dioxane (2.0 mL, 8.0 mmol) in dichloromethane (2 mL) was stirred at room temperature for 1 h. The mixture was basified with saturated aqueous NaHCO3 and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel (0 to 25% MeOH in dichloromethane) to obtain 4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzonitrile. MS: (ES) m / z calcd. C 24 H 27 N4O[M+H] + 387.2, measured value 387.2.

[0190] Step c: A mixture of 4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzonitrile (56.0 mg, 0.15 mmol), 2-bromo-4-(tert-butyl)-1-methylbenzene (131.0 mg, 0.58 mmol), Pd(OAc)2 (12.0 mg, 0.05 mmol), X-Phos (60.0 mg, 0.13 mmol) and CsCO3 (141.0 mg, 0.43 mmol) in dioxane (2 mL) was stirred at 110°C under N2 for 1 h. The mixture was cooled to room temperature, diluted with water, and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 35% EtOAc in hexanes) to give 4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzonitrile. MS: (ES) m / z calculated value C 35 H 41 N4O[M+H] + 533.3, measured value 533.3.

[0191] Step d: To a mixture of 4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzonitrile (36.0 mg, 0.07 mmol) in DCM (1 mL) and DMSO (6 mL) was added 4N aqueous NaOH solution (1.0 mL, 4.0 mmol) and H2O2 (0.40 mL, 35% aqueous solution). The mixture was stirred at room temperature for 30 min, diluted with water, and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 65% EtOAc in hexanes) to give 4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzamide. 1 H NMR (400 MHz, CDCl3): δ 7.65 (dd, J = 8.0, 8.4 Hz, 2H), 7.12–7.24 (m, 5H), 7.05 (dd, J = 8.0, 2.0 Hz, 1H), 6.81 (d, J = 7.2 Hz, 1H), 6.70 (d, J = 8.4 Hz, 1H), 6.05 (br s, 1H), 5.73 (br s, 1H) 4.11 (m, 2H), 3.56 (m, 2H), 3.36 (m, 2H), 3.01 (m, 2H), 2.31 (s, 3H), 2.09 (s, 3H), 1.86 (m, 1H), 1.31 (s, 9H), 0.81 (m, 6H). MS: (ES) m / z calcd. 35 H 43 N4O2[M+H] + 551.3, measured value 551.3.

[0192] Example 3

[0193] Synthesis of 1-(4-(5-(3,5-dichloropyridin-2-yl)-2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

[0194]

[0195] Step a: To a mixture of 2,5-difluoro-4-nitrobenzoic acid (35.5 g, 174.8 mmol) in dichloromethane (400 mL) was slowly added oxalyl chloride (16.0 mL, 186.3 mmol) at room temperature, followed by DMF (0.2 mL, 2.6 mmol). The resulting mixture was stirred at room temperature overnight. It was then concentrated in vacuo to obtain 2,5-difluoro-4-nitrobenzoyl chloride, which was used directly in the next step.

[0196] Step b: MgCl2 (34.0 g, 357.1 mmol), 2,5-difluoro-4-nitrobenzoyl chloride (38.8 g, 175.1 mmol) and triethylamine (50.0 mL, 355.8 mmol) were added to a mixture of tert-butyl 4-oxypiperidine-1-carboxylate (35.6 g, 175.8 mmol) in dichloromethane (500 mL) at 0°C. The mixture was stirred at 0°C for 30 min and then at room temperature for 7 h. It was then cooled to 0°C, quenched with saturated NH4Cl and extracted with dichloromethane. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure to give tert-butyl 3-(2,5-difluoro-4-nitrobenzoyl)-4-oxypiperidine-1-carboxylate. MS: (ES) m / z calculated value C 17 H 19 F2N2 O6Na[M+Na] + 385.1, measured value 385.1.

[0197] Step c: A mixture of tert-butyl 3-(2,5-difluoro-4-nitrobenzoyl)-4-oxopiperidine-1-carboxylate (30.0 g, 78.1 mmol) and (2,6-dimethylphenyl)hydrazine hydrochloride (17.5 g, 101.5 mmol) in EtOH (30 mL) was heated to 75 °C for 4 h. The mixture was then cooled to room temperature, concentrated under reduced pressure, and the residue was purified by silica gel flash chromatography (0 to 30% EtOAc in hexanes) to obtain tert-butyl 3-(2,5-difluoro-4-nitrophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 25 H 26 F2N4O4[M+H] + 485.2, measured value 485.2.

[0198] Step d: A mixture of tert-butyl 3-(2,5-difluoro-4-nitrophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (10.0 g, 20.6 mmol) and PtO2 (0.5 g, 2.2 mmol) in EtOAc (100 mL) was stirred overnight in a Parr shaker under a hydrogen atmosphere at 40 psi. The mixture was filtered through celite and concentrated under vacuum to give tert-butyl 3-(4-amino-2,5-difluorophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 25 H 28 F2N4O2[M+H] + 455.2, measured value 455.2.

[0199] Step e: A mixture of tert-butyl 3-(4-amino-2,5-difluorophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (7.9 g, 17.3 mmol) and benzoyl isocyanate (2.0 g, 17.3 mmol) in THF (40 mL) was stirred at room temperature for 3 h. The mixture was concentrated under reduced pressure to give tert-butyl 3-(4-(3-benzoylureido)-2,5-difluorophenyl-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.

[0200] A mixture of tert-butyl 3-(4-(3-benzoylureido)-2,5-difluorophenyl-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (-17.3 mmol, from the previous step) and K2CO3 (7.1 g, 51.3 mmol) in MeOH (100 mL) was stirred at room temperature for 2 h and then at 50°C for 20 min. The mixture was stirred at room temperature for 2 h and then at 50°C for 20 min. Extracted with IPA:CHCl3 (1:3). The organic layer was separated, dried over Na2SO4, concentrated under reduced pressure, and purified by silica gel flash chromatography (0 to 70% EtOAc in hexanes) to give tert-butyl 3-(2,5-difluoro-4-ureidophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 26 H 29 F2N5O3[M+H] + 498.2, measured value 498.2.

[0201] Step f: A mixture of tert-butyl 3-(2,5-difluoro-4-ureidophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (7.0 g, 14.1 mmol) and 4N HCl in dioxane (14.0 mL, 56.0 mmol) in dichloromethane (20 mL) was stirred at room temperature overnight. The mixture was then concentrated in vacuo to give 1-(4-(2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea hydrochloride. MS: (ES) m / z calculated value C 21 H 21 F2N5O[M+H] + 398.2, measured value 398.2.

[0202] Step g: Triethylamine (3.9 mL, 27.7 mmol) was added to a suspension of 1-(4-(2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea hydrochloride (4 g, 9.23 mmol), 3,5-dichloro-2-fluoropyridine (1.7 g, 10.2 mmol) and Li2CO3 (2.0 g, 27.7 mmol) in DMSO (14 mL) under magnetic stirring. The resulting mixture was stirred at 90 °C for 4 h. After cooling to room temperature, the reaction mixture was diluted with DCM, washed with brine and dried over MgSO4. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (20 to 50% EtOAc in hexanes followed by 0 to 50% EtOAc in dichloromethane) followed by recrystallization in MeOH / dichloromethane / EtOAc to afford l-(4-(5-(3,5-dichloropyridin-2-yl)-2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea. 1 H NMR (400 MHz, CD3OD): δ 8.16 (d, J = 2.3 Hz, 1H), 8.01 (dd, J = 6.6, 12.5 Hz, 1H), 7.83 (d, J = 2.3 Hz, 1H), 7.25 (dd, J = 7.0, 8.2 Hz, 1H), 7.06–7.16 (m, 2H), 6.70 (dd, J = 6.6, 11.4 Hz, 1H), 4.86 (br, 3H), 4.37 (s, 2H), 3.76 (t, J = 5.8 Hz, 2H), 3.03 (t, J = 5.8 Hz, 2H), 1.99 (s, 6H). MS: (ES) m / z calcd. C 26 H23 Cl2F2N6O[M+H] + 543.1, measured value 543.1.

[0203] Example 4

[0204] Synthesis of 1-(4-(2-(2,6-diethylphenyl)-5-(tert-pentyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

[0205]

[0206] WARNING: The formation of diazo compounds is potentially hazardous, please handle with caution and wear appropriate personal protective equipment!

[0207] Step a: Under magnetic stirring, add 2,6-diethylaniline (10.0 g, 67 mmol) to a 250 mL flask containing 90 mL of concentrated hydrochloric acid. Stir the resulting mixture for 30 min and cool it with an ice-salt bath until the internal temperature reaches -5 °C. Slowly add a solution of sodium nitrite (5.5 g, 80 mmol) in water (60 mL) to the above mixture while keeping the internal temperature below 5 °C.

[0208] Separately, tin(II) chloride dihydrate (31.6 g, 140 mmol) was added to a 500 mL 3-neck round bottom flask charged with concentrated hydrochloric acid (60 mL) under mechanical stirring, and the resulting solution was then cooled with an ice bath.

[0209] Afterwards, the diazo slurry was filtered into a 500 ml flask containing a cooled tin chloride solution under vigorous stirring. After 90 min, the reaction mixture was transferred to a 500 ml Erlenmeyer flask and the flask was rinsed with water (20 ml) and chloroform (8 ml). The combined mixture was stirred overnight at room temperature. The entire liquid layer was decanted to obtain a wet solid. The recovered material was dried in a vacuum for one day and then transferred to a 500 ml 3-neck round bottom flask equipped with an overhead mechanical stirrer and stirred with ether (180 ml). The mixture was cooled in an ice bath and NaOH solution (10 N, 30 ml) was slowly added to the mixture while maintaining the internal temperature below 12 ° C. After addition, the mixture was allowed to stand on ice for 2 h. The ether layer was decanted into a 500 ml flask and a stream of hydrogen chloride gas was blown into the ether solution while stirring. The resulting precipitate was collected by filtration to obtain (2,6-diethylphenyl)hydrazine hydrochloride. MS: (ES) m / z calcd. C 10 H 17 N2[M+H] + 165.1, measured value 165.1.

[0210] Step b: A mixture of tert-butyl 3-(2,5-difluoro-4-nitrobenzoyl)-4-oxypiperidine-1-carboxylate (60.0 g, 156.1 mmol) and (2,6-diethylphenyl) hydrazine hydrochloride (30.0 g, 149.5 mmol) in EtOH (560 mL) was heated to 50 ° C for 3 h. The mixture was then cooled to room temperature and quenched with saturated NaHCO3 aqueous solution. The mixture was extracted with EtOAc, the organic layer was separated, washed with brine, dried with Na2SO4 and filtered. The solvent was concentrated under reduced pressure, and the residue was purified by silica gel flash chromatography (0 to 50% EtOAc in hexane) to obtain tert-butyl 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calcd. C 27 H 31 F2N4O4[M+H] + 513.2, measured value 513.5.

[0211] Step c: A mixture of tert-butyl 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (9.6 g, 18.7 mmol), iron powder (10.0 g, 179.1 mmol) and NH4Cl (15.0 g, 280.4 mmol) in EtOH (200 mL) and water (20 mL) was heated to 80 ° C for 1 h. The mixture was then cooled to room temperature, filtered through celite, and washed with EtOAc. The filtrate was partitioned between EtOAc and saturated NaHCO3. The organic layer was separated, dried over Na2SO4 and filtered. The solvent was concentrated under vacuum to give tert-butyl 3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 27 H 33 F2N4O2[M+H] + 483.3, measured value 483.3.

[0212] Step d: A mixture of tert-butyl 3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (9.2 g, 19.1 mmol) and benzoyl isocyanate (11.3 g, 76.8 mmol) in THF (100 mL) was stirred at room temperature for 4 h. The mixture was then concentrated in vacuo to give tert-butyl 3-(4-(3-benzoylureido)-2,5-difluorophenyl-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.

[0213] A mixture of tert-butyl 3-(4-(3-benzoylureido)-2,5-difluorophenyl-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (-19.1 mmol) from the previous step and K2CO3 (10.0 g, 72.3 mmol) in MeOH (100 mL) was stirred at room temperature for 7 h. The reaction mixture was extracted with IPA:CHCl3 (1:3). The organic phase was separated. The layer was dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by flash chromatography on silica gel (0 to 100% EtOAc in dichloromethane, followed by 0 to 20% MeOH in dichloromethane) to give tert-butyl 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 28 H 34 F2N5O3[M+H] + 526.3, measured value 526.3.

[0214] Step e: A mixture of tert-butyl 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (2.9 g, 5.5 mmol) and 4N HCl in dioxane (35.0 mL, 140.0 mmol) in dichloromethane (30 mL) was stirred at room temperature for 1.5 h. The mixture was then concentrated in vacuo to give 1-(4-(2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea hydrochloride. MS: (ES) m / z calculated value C 23 H 26 F2N5O[M+H] + 426.2, measured value 426.2.

[0215] Step f: To a mixture of 1-(4-(2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea hydrochloride (115.0 mg, 0.3 mmol) in THF (2.5 mL) at 0°C, 3-chloro-3-methylbut-1-yne (32 mg, 0.3 mmol), NEt3 (88 μL, 0.6 mmol) and CuCl (30 mg, 0.3 mmol) were added sequentially. The mixture was warmed to room temperature and stirred for 30 minutes. The mixture was then basified with saturated aqueous NaHCO3 and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 100% EtOAc in hexanes) to give 1-(4-(2-(2,6-diethylphenyl)-5-(2-methylbut-3-yn-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea. MS: (ES) m / z calculated value C 28 H 32 F2N5O[M+H] + 492.3, measured value 492.2.

[0216] Step g: A mixture of 1-(4-(2-(2,6-diethylphenyl)-5-(2-methylbut-3-yn-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea (66.0 mg, 0.13 mmol) and Pd / C (50 mg, 50% aqueous solution) in EtOAc (35 mL) was stirred in a Parr shaker for 1.5 h under a hydrogen atmosphere at 53 psi. The mixture was filtered through celite. The filtrate was concentrated under reduced pressure and the residue was purified by HPLC (MeCN / H2O with 0.1% TFA), basified with saturated aqueous NaHCO3 and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4, filtered and treated with 1.0 M HCl in diethyl ether. The solvent was concentrated in vacuo to give l-(4-(2-(2,6-diethylphenyl)-5-(tert-pentyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea hydrochloride. 1HNMR (400MHz, CD3OD): δ8.15(br s,1H),7.47(dd,J=7.4,7.4Hz,1H),7.29(m,2H),6.68(brs,1H),4.57(br s,2H),4.18(br s,1H),3.62(br s,1H),3.32(br s,2H),2.38(br s,2H),2.22(br s,2H),1.98(br s,2H),1.70(s,1H),1.55(br s,6H),1.55(br s,6H),1.32–1.42(m,3H),1.00–1.22(m,9H).MS:(ES)m / z calculated value C 28 H 36 F2N5O[M+H] + 496.3, measured value 496.3.

[0217] Example 5

[0218] Synthesis of N-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)formamide

[0219]

[0220] Step a: A mixture of tert-butyl 3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (1.7 g, 3.5 mmol), acetyl chloride (1.4 mL, 19.6 mmol) and NEt3 (3.7 mL, 26.1 mmol) in THF (120 mL) was stirred at room temperature for 2 h. The reaction mixture was quenched with saturated NaHCO3 aqueous solution and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 80% EtOAc in hexanes) to give tert-butyl 3-(4-acetamido-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 29 H 35 F2N4O3[M+H] + 525.3, measured value 525.6.

[0221] Step b: A mixture of tert-butyl 3-(4-acetamido-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (1.3 g, 2.4 mmol) and 4N HCl in dioxane (20.0 mL, 80.0 mmol) in dichloromethane (20 mL) was stirred at room temperature for 0.5 h. The mixture was then concentrated in vacuo to give N-(4-(2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)acetamide hydrochloride. MS: (ES) m / z calculated value C 24 H 27 F2N4O[M+H] + 425.2, measured value 425.2.

[0222] Step c: A mixture of N-(4-(2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)acetamide hydrochloride (0.45 g, 1.0 mmol), 2,4-bis(trifluoromethyl)benzaldehyde (0.7 g, 2.9 mmol), NaBH(OAc)3 (0.8 g, 3.8 mmol), NEt3 (0.5 mL, 3.6 mmol) and HOAc (0.2 mL, 3.3 mmol) in dichloromethane (5 mL) was stirred at 30 °C for 1 h. The reaction was quenched with saturated aqueous NaHCO3 and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 80% EtOAc in hexanes) to give N-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)acetamide. MS: (ES) m / z calculated value C 33 H 30 F8N4O[M+H] + 651.3, measured value 651.6.

[0223] Step d: A mixture of N-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)acetamide (0.5 g, 0.8 mmol) and 4N HCl in dioxane (3.0 mL, 12.0 mmol) in water (0.6 mL) was stirred at 80 °C for 40 minutes. The mixture was cooled to room temperature, basified with saturated aqueous NaHCO3, and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 70% EtOAc in hexanes) to give 4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluoroaniline. MS: (ES) m / z calculated value C 31 H 29 F8N4[M+H] + 609.2, measured value 609.2.

[0224] Step e: A mixture of 4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluoroaniline (25.0 mg, 0.04 mmol) and HCO2H (1 mL) was stirred at 75°C for 40 minutes. The mixture was cooled to room temperature, basified with saturated aqueous NaHCO3, and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 70% EtOAc in hexanes) to give N-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)formamide. 1H NMR (400 MHz, CDCl3): δ 8.38 (d, J = 1.2 Hz, 1H), 8.15 (m, 2H), 7.89 (br s, 1H), 7.79 (d, J = 8.4 Hz, 1H), 7.53 (br s, 1H), 7.28 (dd, J = 7.6, 7.6 Hz, 1H), 7.10 (d, J = 7.6 Hz, 2H), 6.55 (m, 1H), 3.95 (s, br, 2H), 3.58 (bs, 2H), 2.93 (m, 4H), 2.31 (sextet, J = 7.6 Hz, 2H), 2.19 (sextet, J = 7.2 Hz, 2H), 1.08 (t, J = 7.4 Hz, 6H). MS: (ES) m / z calculated value C 32 H 29 F8N4O[M+H] + 637.2, measured value 637.2.

[0225] Example 6

[0226] Synthesis of 1-(4-(2-(2,6-diethylphenyl)-5-(2-hydroxy-2-methyl-1-phenylpropyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

[0227]

[0228] Step a: A mixture of tert-butyl 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (1.0 g, 2.0 mmol) and 4N HCl in dioxane (5.0 mL, 20.0 mmol) in dichloromethane (10 mL) was stirred at room temperature for 1.5 h. The mixture was then concentrated in vacuo to give 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine hydrochloride. MS: (ES) m / z calculated value C 22 H 23 F2N4O2[M+H] + 413.2, measured value 413.2.

[0229] N,N-diisopropylethylamine (3.0 mL, 17.3 mmol) was added to a suspension of the above 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine hydrochloride (~2.0 mmol) and ethyl 2-bromo-2-phenylacetate (2.0 mL, 11.4 mmol) in acetonitrile (6 mL) under magnetic stirring. The resulting mixture was stirred at 90 °C for 3 h. After cooling to room temperature, the reaction mixture was diluted with EtOAc, washed with brine and dried over MgSO4. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (10% to 30% EtOAc in hexanes) to give ethyl 2-(2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate. MS: (ES) m / z calculated value C 32 H 33 F2N4O4[M+Na] + 575.2, measured value 575.3.

[0230] Step b: A mixture of ethyl 2-(2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate (1.0 g, 1.7 mmol), iron powder (1.0 g, 17.9 mmol) and NH4Cl (1.5 g, 26.9 mmol) in EtOH (20 mL) and water (2 mL) was heated to 80 °C for 1 h. The mixture was then cooled to room temperature, filtered through celite, and washed with EtOAc. The filtrate was partitioned between EtOAc and saturated aqueous NaHCO3. The organic layer was separated, dried over Na2SO4 and filtered. The solvent was concentrated under vacuum to give ethyl 2-(3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate. MS: (ES) m / z calculated value C 32 H 35 F2N4O2[M+H] + 545.3, measured value 545.3.

[0231] Step c: A mixture of ethyl 2-(3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate (800.0 mg, 1.5 mmol) and benzoyl isocyanate (1.1 g, 7.7 mmol) in THF (10 mL) was stirred at room temperature for 4 h. The mixture was then concentrated in vacuo to give ethyl 2-(3-(4-(3-benzoylureido)-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate.

[0232] A mixture of ethyl 2-(3-(4-(3-benzoylureido)-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate (-1.5 mmol) from the previous step and KCO (1.0 g, 7.2 mmol) in MeOH (10 mL) was stirred at room temperature overnight. The reaction mixture was extracted with EtOAc. The organic layer was separated, dried over NaSO and filtered. The solvent was concentrated under reduced pressure and the residue was purified by flash chromatography on silica gel (10 to 45% EtOAc in hexanes) to give ethyl 2-(2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate. MS: (ES) m / z calcd. C 33 H 36 F2N5O3[M+H] + 588.3, ​​measured value 588.3.

[0233] Step d: To a solution of ethyl 2-(2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate (100.0 mg, 0.2 mmol) in THF (5 mL) was added 1.6 M CH3Li in ether (0.7 mL, 1.1 mmol) at 0°C. The resulting mixture was stirred at the same temperature for 30 min, quenched with saturated NH4Cl, and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by preparative TLC (40% EtOAc in hexanes followed by 30% EtOAc in dichloromethane) to give 1-(4-(2-(2,6-diethylphenyl)-5-(2-hydroxy-2-methyl-1-phenylpropyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea. 1 H NMR (400 MHz, CD3OD): δ7.93 (dd, J = 6.6, 12.4 Hz, 1H), 7.10-7.50 (m, 8H), 6.54 (dd, J = 6.6, 11.6 Hz, 1H), 4.86 (br, 3H), 3.41-3.67 (m, 4H), 2.63-2.92 (m, 3H), 2.02-2.32 (m, 4H), 1.32 (s, 3H), 1.28 (br s, 1H), 1.16 (s, 3H), 1.10 (t, J = 7.8 Hz, 3H), 1.01 (t, J = 7.8 Hz, 3H). MS: (ES) m / z calculated value C 33 H 38 F2N5O2[M+H] + 574.3, measured value 574.5.

[0234] Example 7

[0235] Synthesis of 1-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

[0236]

[0237] Step a: To a mixture of tert-butyl 3,3-dimethyl-4-oxypiperidine-1-carboxylate (4.0 g, 17.6 mmol) in DCM (50 mL) at 0°C, MgCl2 (3.4 g, 35.2 mmol), 2,5-difluoro-4-nitrobenzoyl chloride (4.3 g, 19.4 mmol) and triethylamine (4.9 mL, 35.2 mmol) were added in sequence. The mixture was stirred at ℃ for 30 min and then stirred at room temperature for 1.5 h. The mixture was then cooled to 0°C, quenched with saturated NH4Cl and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure to give tert-butyl 5-(2,5-difluoro-4-nitrobenzoyl)-3,3-dimethyl-4-oxypiperidine-1-carboxylate. MS: (ES) m / z calculated value C 19 H 22 F2N2O6Na[M+Na] + 435.1, measured value 435.1.

[0238] Step b: A mixture of tert-butyl 5-(2,5-difluoro-4-nitrobenzoyl)-3,3-dimethyl-4-oxopiperidine-1-carboxylate (-17.6 mmol, crude from step a), (2,6-diethylphenyl)hydrazine hydrochloride (3.5 g, 17.6 mmol) and pyridine (2.1 mL, 26.4 mmol) in MeOH (100 mL) was heated to 45 °C overnight. The mixture was cooled to room temperature and water was added. The mixture was extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 60% EtOAc in hexanes) to give tert-butyl 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 29 H 35 F2N4O4[M+H] + 541.3, measured value 541.6.

[0239] Step c: A mixture of tert-butyl 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (6.7 g, 6.2 mmol), iron powder (12.0 g, 214.9 mmol) and NH4Cl (40.0 g, 742.1 mmol) in EtOH (100 mL) and water (10 mL) was heated to 90 ° C for 30 minutes. The mixture was cooled to room temperature, filtered through celite, and washed with EtOAc. The filtrate was partitioned between EtOAc and saturated aqueous NaHCO3. The organic layer was separated, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by flash chromatography on silica gel (0 to 80% EtOAc in hexanes, then 0 to 30% EtOAc in dichloromethane) to give tert-butyl 3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 29 H 37 F2N4O2[M+H] + 511.3, measured value 511.6.

[0240] Step d: A mixture of tert-butyl 3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (1.0 g, 2.0 mmol) and benzoyl isocyanate (0.5 g, 3.4 mmol) in THF (6 mL) was stirred at room temperature for 1 h. The mixture was concentrated under reduced pressure on a rotary evaporator to give tert-butyl 3-(4-(3-benzoylureido)-2,5-difluorophenyl-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.

[0241] A mixture of tert-butyl 3-(4-(3-benzoylureido)-2,5-difluorophenyl-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (-2.0 mmol) from the previous step and KCO (1.0 g, 7.2 mmol) in MeOH (20 mL) was stirred at room temperature for 2 h. The mixture was washed with IPA:CHCl (1: 3) Extraction. The organic layer was separated, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure, and the residue was purified by silica gel flash chromatography (0 to 80% EtOAc in dichloromethane) to give 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester. MS: (ES) m / z calculated value C 30 H 38 F2N5O3[M+H] + 554.3, measured value 554.2

[0242] Step e: A mixture of tert-butyl 2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (0.6 g, 1.1 mmol) and 4N HCl in dioxane (30.0 mL, 120.0 mmol) in dichloromethane (10 mL) was stirred at room temperature for 1.5 h. The mixture was concentrated under reduced pressure to give 1-(4-(2-(2,6-diethylphenyl)-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea hydrochloride. MS: (ES) m / z calculated value C 25 H 30 F2N5O[M+H] + 454.2, measured value 454.2.

[0243] Step f: To a mixture of 1-(4-(2-(2,6-diethylphenyl)-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea hydrochloride (100.0 mg, 0.2 mmol), 2,4-bis(trifluoromethyl)benzaldehyde (100.0 mg, 0.8 mmol) and N,N-diisopropylethylamine (0.3 mL, 1.7 mmol) in dichloromethane (10 mL) was added NaBH(OAc)3 (250.0 mg, 1.2 mmol) followed by AcOH (2 drops) under magnetic stirring. The resulting mixture was stirred at 45 °C for 3 h. After cooling to room temperature, the reaction mixture was quenched with MeOH, diluted with EtOAc, washed with brine and dried over MgSO4. The solvent was removed under reduced pressure and the residue was purified by preparative TLC (50% EtOAc in hexanes) followed by HPLC (MeCN / H2O with 0.1% TFA) to give 1-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea. 1 H NMR (400 MHz, CD3OD): δ 8.30 (d, J = 8.2 Hz, 1H), 7.83–7.95 (m, 3H), 7.36 (t, J = 7.5 Hz, 1H), 7.20 (d, J = 7.7 Hz, 2H), 6.50 (dd, J = 6.5, 11.6 Hz, 1H), 4.87 (br, 3H), 4.00 (s, 2H), 3.57 (s, 2H), 2.79 (s, 2H), 2.22–2.78 (m, 4H), 1.35 (s, 6H), 1.08 (t, J = 7.8 Hz, 6H). MS: (ES) m / z calculated value C 34 H 34 F8N5O[M+H] + 680.3, measured value 680.6.

[0244] Example 8

[0245] Synthesis of 1-(4-(2-(2,6-diethylphenyl)-5-isobutyryl-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

[0246]

[0247] A mixture of 1-(4-(2-(2,6-diethylphenyl)-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea hydrochloride (25 mg, 0.05 mmol), isobutyric acid (80.0 mg, 1.1 mmol), HATU (100 mg, 0.26 mmol) and NEt3 (0.1 mL, 0.71 mmol) in DMF (1.5 mL) was stirred at 50-70 ° C for 40 min. The mixture was cooled to room temperature, basified with saturated aqueous NaHCO3 solution, and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 100% EtOAc in hexanes) to give 1-(4-(2-(2,6-diethylphenyl)-5-isobutyryl-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea. 1 H NMR (400 MHz, CDCl3): δ 8.06 (m, 1H), 7.38 (dd, J = 7.6, 7.6 Hz, 1H), 7.21 (d, J = 8.0 Hz, 2H), 6.55 (m, 1H), 4.58 (s, 2H), 3.30 (m, 3H), 2.95 (s, 2H), 2.92 (m, 1H), 2.24 (q, J = 7.6 Hz, 4H), 1.58 (s, 6H), 1.04 (m, 12H). MS: (ES) m / z calculated value C 29 H 36 F2N5O2[M+H] + 524.3, measured value 524.6.

[0248] Example 9

[0249] Synthesis of 1-(4-(5-(2-chloro-4-(trifluoromethyl)-phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)urea

[0250]

[0251] Step a: At -78 ° C, in a N2 atmosphere, a solution of tert-butyl 4-oxopiperidin-1-carboxylate (5.0 g, 25.1 mmol) in anhydrous THF (30 mL) was added dropwise with 1.0 M LiHMDS in THF (27.5 mL, 27.5 mmol). After the solution was stirred for 30 min, a solution of 3-fluoro-4-nitrobenzoyl chloride (5.1 g, 25.1 mmol) in THF (6 mL) was added to the mixture. The reaction mixture was stirred at -78 ° C for 1 h, then warmed to room temperature and stirred for 2 h. After completion, the reaction mixture was quenched with 1 M NaHSO4 (50 mL), stirred for 10 min, and then diluted with EtOAc. The organic layer was washed with H2O, brine, dried with Na2SO4, and filtered. The solvent was concentrated under vacuum, and the crude material was used directly in the next step without further purification.

[0252] Step b: At room temperature, to a stirred solution of tert-butyl 3-(3-fluoro-4-nitrobenzoyl)-4-oxypiperidine-1-carboxylate (6.0 g, 16.4 mmol) in EtOH (120 mL) and glacial acetic acid (12.0 mL, 207.9 mmol), (2-isobutoxy-6-methylphenyl) hydrazine hydrochloride (3.8 g, 16.4 mmol) was added. The mixture was stirred for 15 min and then refluxed for 3 h. After the reaction was completed, the solvent was removed under reduced pressure and the residue was diluted with EtOAc (100 mL). The organic layer was washed with 2N NaOH aqueous solution, brine, dried with Na2SO4, and filtered. The solvent was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (5 to 20% dichloromethane in MeOH) to give tert-butyl 3-(3-fluoro-4-nitrophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calcd. C 28 H 34 FN4O5[M+H] + 525.2, measured value 525.3

[0253] Step c: 10% Pd / C (250 mg) was added to a solution of tert-butyl 3-(3-fluoro-4-nitrophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (0.8 g, 1.5 mmol) in methanol (20 mL) at room temperature. The resulting mixture was stirred in hydrogen (30 psi) for 1 h. The reaction mixture was filtered through a pad of celite and the filtrate was concentrated under reduced pressure to give tert-butyl 3-(4-amino-3-fluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate, which was used directly in the next step without further purification.

[0254] Step d: To a solution of tert-butyl 3-(4-amino-3-fluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (0.7 g, 1.4 mmol) from the previous step in anhydrous THF (10 mL) was added K2CO3 (0.8 g, 5.7 mmol) and acetyl chloride (0.4 g, 5.1 mmol) at room temperature. The reaction mixture was stirred at room temperature for 4 h and diluted with water. The mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum to give tert-butyl 3-(4-acetamido-3-fluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.

[0255] Step e: To a solution of tert-butyl 3-(4-acetamido-3-fluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (0.8 g, 1.5 mmol) in dichloromethane (10 mL) was added TFA (0.4 g, 3.6 mmol). The resulting mixture was stirred at room temperature for 2 h. After the reaction was complete, the solvent was diluted with water and saturated aqueous NaHCO3 and extracted with dichloromethane. The organic layer was washed with brine and dried over Na2SO4. The solvent was removed under reduced pressure to give N-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)acetamide. MS: (ES) m / z calculated value C 25 H 30 FN4O2[M+H] + 437.2, measured value 437.3.

[0256] Step f: To a mixture of N-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)acetamide (100.0 mg, 0.25 mmol), 1-bromo-2-chloro-4-(trifluoromethyl)benzene (85.0 mg, 0.37 mmol), NaOtBu (47.0 mg, 0.49 mmol) and BINAP (80.0 mg, 0.05 mmol) in toluene (3 mL) was added Pd2(dba)3 (22 mg, 0.02 mmol). The reaction mixture was degassed (N2) for 5 min and stirred at 105 °C in N2 for 6 h. After completion of the reaction, the mixture was cooled to room temperature, diluted with EtOAc and filtered through celite. The filtrate was washed with brine and dried over Na2SO4. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (5 to 20% EtOAc in hexanes) to give N-(4-(5-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)acetamide. MS: (ES) m / z calculated value C 32 H 32 ClF4N4O2[M+H] + 615.2, measured value 615.3.

[0257] Step g: To a solution of N-(4-(5-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)acetamide (100.0 mg, 0.4 mmol) in MeOH (3 mL) was added 4N HCl in dioxane (2.5 mL, 10.0 mmol). The resulting mixture was stirred at room temperature for 2 h. After the reaction was complete, the mixture was diluted with saturated aqueous NaHCO3 and extracted with dichloromethane. The combined organic layers were washed with brine and dried over Na2SO4. The solvent was removed under reduced pressure to give 4-(5-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluoroaniline. MS: (ES) m / z calculated value C 30 H 30 ClF4N4O[M+H] + 573.2, measured value 573.2.

[0258] Step g: To a stirred solution of 4-(5-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluoroaniline (75.0 mg, 0.13 mmol) in anhydrous THF (5 mL) was added N,N-diisopropylethylamine (75.0 mg, 0.65 mmol) and trimethyl isocyanate (140.0 mg, 1.2 mmol). The reaction mixture was stirred at room temperature for 16 h. Upon completion, the reaction mixture was diluted with EtOAc, washed with brine and dried over Na2SO4. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (20 to 60% EtOAc in hexanes) to give 1-(4-(5-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)urea. 1 H NMR (400MHz, CD3OD): δ8.00(t,J=8.6Hz,1H),7.69(d,J=2.0Hz,1H),7.54(d,J=8.6Hz, 1H),7.39(d,J=8.2Hz,1H),7.33(t,J=7.8Hz,1H),7.0(d,J=8.2Hz,1H),6.80–6.95(m,3 H), 4.87 (br, 3H), 4.32 (q, J = 9.4 Hz, 2H), 3.65–3.75 (m, 2H), 3.61 (t, J = 5.8 Hz, 2H), 2.95–3.10 (m, 2H), 2.01 (s, 3H), 1.85–1.98 (m, 1H), 0.86 (d, J = 6.6 Hz, 6H). MS: (ES) m / z calculated value C 31 H 31 ClF4N5O2[M+H] + 616.2, measured value 616.2.

[0259] Example 10

[0260] Synthesis of 1-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-5-((2-(trifluoromethyl)phenyl)sulfonyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)urea

[0261]

[0262] Step a: A mixture of tert-butyl 3-(4-amino-3-fluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (2.4 g, 4.9 mmol) and trimethylsilyl isocyanate (7.5 g, 65.6 mmol) and acetic acid (2.87 mL, 47.8 mmol) in dichloromethane (60 mL) was stirred at room temperature for 8 h. The mixture was then basified with saturated aqueous NaHCO3 and extracted with dichloromethane. The organic layer was separated, washed with brine, dried over Na2SO4 and filtered. The solvent was concentrated under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 100% EtOAc in hexanes) to give tert-butyl 3-(3-fluoro-4-ureidophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 29 H 37 FN5O4[M+H] + 538.3, measured value 538.3.

[0263] Step b: A mixture of tert-butyl 3-(3-fluoro-4-ureidophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (1.8 g, 3.3 mmol) and 4N HCl in dioxane (17.0 mL, 68.0 mmol) in dichloromethane (20 mL) was stirred at room temperature for 1 h. The mixture was then concentrated in vacuo to give 1-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)urea hydrochloride. MS: (ES) m / z calculated value C 24 H 29 FN5O2[M+H] + 438.2, measured value 438.3

[0264] Step c: A mixture of 1-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)urea hydrochloride (25.0 mg, 0.05 mmol), 2-(trifluoromethyl)benzenesulfonyl chloride (30.0 mg, 0.12 mmol) and NEt3 (0.1 mL, 0.7 mmol) in dichloromethane (1.5 mL) was stirred at room temperature for 30 min. The mixture was quenched with water. The resulting mixture was purified by silica gel flash chromatography (0 to 100% EtOAc in hexanes) to afford l-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-5-((2-(trifluoromethyl)phenyl)sulfonyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)urea. 1 H NMR (400MHz, CDCl3): δ8.17(m,1H),7.97(dd,J=8.4,8.4Hz,1H),7.89(m,1H), 7.70(m,2H),7.18(dd,J=8.0,8.0Hz,1H),7.07(d,J=2.8Hz,1H),6.68-6.84(m, 4H),4.87(s,2H),4.50(q,J=13.6Hz,2H),3.73(m,2H),3.58(d,J=6.8Hz,2H), 2.92(m,2H),1.94(s,3H),1.85(m,1H),0.78(d,J=6.8Hz,6H).MS:(ES)m / z calculated value C 31 H 32 F4N5O4S[M+H] + 646.2, measured value 646.2.

[0265] Embodiment 11

[0266] Synthesis of 4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenol

[0267]

[0268] Step a: At -78 ° C, in a N2 atmosphere, a solution of tert-butyl 4-oxopiperidin-1-carboxylate (3.0 g, 15.1 mmol) in anhydrous THF (30 mL) was added dropwise with 1.0 M LiHMDS in THF (16.5 mL, 16.5 mmol). After the reaction mixture was stirred for 30 min, a solution of 4-methoxybenzoyl chloride (2.6 g, 15.1 mmol) in THF (3 mL) was added to the mixture. The reaction mixture was stirred at -78 ° C for 1 h, after which the mixture was warmed to room temperature and stirred for 2 h. After completion, the reaction mixture was diluted with EtOH: AcOH (3: 1), and (2,6-dimethylphenyl) hydrazine hydrochloride (2.6 g, 15.1 mmol) was added. The mixture was stirred for 30 min, and then stirred at 100 ° C for 16 h. After the reaction was completed, the mixture was cooled to room temperature and diluted with EtOAc. The organic layer was washed with H2O and then with brine. The combined organic layers were dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel flash chromatography (5 to 20% EtOAc in hexanes) to give tert-butyl 2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate. MS: (ES) m / z calculated value C 26 H 32 N3O3[M+H] + 434.2, measured value 434.3

[0269] Step b: TFA (1.8 g, 16.1 mmol) was added to a solution of tert-butyl 2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate (3.5 g, 8.1 mmol) in dichloromethane (30 mL). The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the solvent was diluted with water and saturated aqueous NaHCO3 and extracted with dichloromethane. The organic layer was washed with brine and dried over Na2SO4. The solvent was evaporated in vacuo to give 2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine. MS: (ES) m / z calculated value C 21 H 24 N3O[M+H] + 334.2, measured value 334.2.

[0270] Step c: Pd(OAc)2 (17.0 mg, 0.07 mmol) was added to a mixture of 2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine (500.0 mg, 1.5 mmol), 4-tert-butyl-2-bromo-1-methylbenzene (502.0 mg, 2.2 mmol), NaOtBu (280.0 mg, 2.9 mmol) and X-Phos (70.0 mg, 2.2 mmol) in toluene (10 mL). The reaction mixture was degassed (N2) for 5 min and stirred at 110 °C in N2 for 16 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with EtOAc, and then filtered through celite. The filtrate was washed with brine and dried over Na2SO4. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (5 to 20% EtOAc in hexanes) to give 5-(5-(tert-butyl)-2-methylphenyl)-2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine. 1 H NMR (400 MHz, CD3OD): δ7.22 (m, 1H), 7.18 (d, J = 1.9 Hz, 1H), 7.13 (bs, 1H), 7.10 (d, J = 2.7 Hz, 1H), 7.09 (s, 1H), 7.05–7.07 (m, 2H), 7.0–7.02 (m, 1H), 6.83 (d, J = 2.4 Hz, 1H), 6.81 (d, J = 2.4 Hz, 1H), 4.13 (s, 2H), 3.73 (s, 3H), 3.38 (t, J = 5.8 Hz, 2H), 2.89 (t, J = 5.8 Hz, 2H), 2.29 (s, 3H), 1.98 (s, 6H), 1.26 (s, 9H). MS: (ES) m / z calcd. C 32 H 38 N3O[M+H] + 480.3, measured value 480.3.

[0271] Step d: To a stirred solution of 5-(5-(tert-butyl)-2-methylphenyl)-2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine (400.0 mg, 0.8 mmol) in anhydrous dichloromethane (10 mL) was added dropwise 1.0 M BBr3 in dichloromethane (2.1 mL, 2.1 mmol) at -78 °C under N2 atmosphere. The reaction mixture was stirred at -78 °C for 1 h, then warmed to room temperature and stirred for 2 h. Upon completion, the reaction mixture was quenched with MeOH (2 mL), stirred for 10 min, and then diluted with dichloromethane. The organic layer was washed with H2O and brine. The organic layer was dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel flash chromatography (5 to 25% EtOAc in hexanes) to afford 4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenol. 1 H NMR (400 MHz, CD3OD): δ7.49 (bs, 1H), 7.25–7.35 (m, 4H), 7.14 (d, J = 7.4 Hz, 2H), 6.98 (d, J = 2.0 Hz, 1H), 6.96 (d, J = 2.0 Hz, 1H), 6.69 (d, J = 2.4 Hz, 1H), 6.67 (d, J = 2.4 Hz, 1H), 4.69 (bs, 2H), 3.80–3.90 (m, 2H), 3.12 (bt, J = 7.0 Hz, 2H), 2.44 (s, 3H), 1.99 (s, 6H), 1.30 (s, 9H). MS: (ES) m / z calcd. C 31 H 36 N3O[M+H] + 466.3, measured value 466.3.

[0272] Example 12

[0273] This example illustrates the evaluation of biological activity associated with specific compounds of the present invention.

[0274] Materials and Methods

[0275] A. Cells

[0276] 1. Cells expressing C5a receptor

[0277] a) U937 cells

[0278] U937 cells are a monocytic cell line expressing C5aR and are available from ATCC (VA). These cells were cultured as a suspension in RPMI-1640 medium supplemented with 2 mM L-glutamine, 1.5 g / L sodium bicarbonate, 4.5 g / L glucose, 10 mM HEPES, 1 mM sodium pyruvate, and 10% FBS. The cells were grown at 37°C in 5% CO2 / 95% air, 100% humidity, and subcultured twice a week at 1:6 (cells were cultured at 1x10 5 to 2x10 6 cells / mL) and cultured at a density of 1x10 6 Cells / mL were harvested. Prior to the assay, cells were treated with 0.5 mM cyclic AMP (Sigma, OH) overnight and washed once before use. U937 cells treated with cAMP can be used for C5aR ligand binding and functional analysis.

[0279] b) Isolated human neutrophils

[0280] Human or mouse neutrophils can be optionally used for the analysis of compound activity. Neutrophils can be separated from fresh human blood using density separation and centrifugal separation. In short, whole blood is incubated with 3% dextran in equal parts for 45 minutes, and separated. After separation, the top layer is laid on the top of 15 milliliters of Ficoll (30 milliliters of blood suspension corresponding to every 15 milliliters of Ficoll), and centrifuged for 30 minutes without braking at 400x g. The precipitation at the bottom of the test tube is then separated and resuspended in PharmLyse RBC lysis buffer (BD Biosciences (BD Biosciences), San Jose, California), and then the sample is centrifuged for 10 minutes with 400x g braking again. The remaining cell pellet is suitably resuspended, and it is composed of the neutrophil granulocytes separated.

[0281] B. Analysis

[0282] 1. Inhibition of C5aR ligand binding

[0283] U937 cells expressing C5aR treated with cAMP were centrifuged and resuspended in assay buffer (20 mM HEPES pH 7.1, 140 mM NaCl, 1 mM CaCl2, 5 mM MgCl2 and 0.1% bovine serum albumin) to a concentration of 3 × 10 6 The binding assay was set up as follows. 0.1 mL of cells were added to the assay plate containing 5 μL of compound, giving a final concentration of approximately 2-10 μM for each compound screened (or partial dose response for compound IC 50 Then, add 0.1 mL 125I-labeled C5a (obtained from Perkin Elmer Life Sciences, Boston, MA) was diluted in assay buffer to a final concentration of 50 pM, yielding 30,000 cpm / well, and the plates were sealed and incubated on a shaker platform at 4°C for approximately 3 hours. Reactions were aspirated onto GF / B glass filters pre-soaked in a 0.3% polyethyleneimine (PEI) solution on a vacuum cell harvester (Packard Instruments; Meriden, CT). Scintillation fluid (40 μl; Microscint 20, Packard Instruments) was added to each well, the plates were sealed, and radioactivity was measured in a Top Count scintillation counter (Packard Instruments). Control wells containing either diluent alone (total counts) or excess C5a (1 μg / mL for nonspecific binding) were used to calculate the total percent inhibition of the compounds. IC was calculated using the computer program Prism from GraphPad, Inc. (San Diego, CA). 50 Value. IC 50 Values ​​are those concentrations required to reduce the binding of radiolabeled C5a to the receptor by 50%. (For further description of ligand binding and other functional assays, see Dairaghi et al., J. Biol. Chem. 274:21569-21574 (1999), Penfold et al., Proc. Natl. Acad. Sci. USA. 96:9839-9844 (1999), and Dairaghi et al., J. Biol. Chem. 272:28206-28209 (1997)).

[0284] 2. Calcium mobilization

[0285] Optionally, the ability of the compound to inhibit calcium flux in the cell can be further determined. To detect the release of intracellular calcium stores, cells (e.g., cAMP-stimulated U937 or neutrophils) are incubated with 3 μM INDO-1AM dye (Molecular Probes; Eugene, OR) in cell culture medium at room temperature for 45 minutes and then washed with phosphate-buffered saline (PBS). After INDO-1AM loading, the cells are resuspended in flux buffer (Hank's balanced salt solution (HBSS) and 1% FBS). Calcium mobilization is measured using a Photon Technology International spectrophotometer (Photon Technology International; New Jersey) with excitation at 350 nm and dual simultaneous recording of fluorescence emission at 400 nm and 490 nm. Relative intracellular calcium levels are expressed as the 400 nm / 490 nm emission ratio. Experiments were performed at 37°C with 10 6 Cells were added in 2 mL of flow buffer with constant mixing. Chemokine ligands can be used in the range of 1 to 100 nM. The emission ratio was plotted over time (typically 2-3 minutes). Candidate ligand blocking compounds (up to 10 μM) were added at 10 seconds followed by chemokines (i.e., C5a; R&D Systems; Minneapolis, MN) at 60 seconds and control chemokines (i.e., SDF-1α; R&D Systems; Minneapolis, MN) at 150 seconds.

[0286] 3. Chemotaxis assay

[0287] Optionally, the ability of the compound to inhibit chemotaxis in cells can be further analyzed. Chemotaxis analysis was performed using 5 μm pore polycarbonate, in a polyvinylpyrrolidone-coated filter in a 96-well chemotaxis chamber (Neuroprobe; Gaithersburg, Maryland) using chemotaxis buffer (Hanks balanced salt solution (HBSS) and 1% FBS). C5aR ligands (i.e., C5a, R&D systems; Minneapolis, Minnesota) were used to assess compound-mediated inhibition of C5aR-mediated migration. Other chemokines (i.e., SDF-1α; R&D systems; Minneapolis, Minnesota) were used as specific controls. The lower chamber was filled with 29 μl of chemokines (i.e., 0.03 nM C5a) and different amounts of compounds; 20 μl of 100,000 U937 or neutrophils were contained in the top chamber. The chambers were incubated at 37°C for 1.5 h, and the number of cells in the lower chamber was quantified by direct cell counting in five high-power fields per well or by CyQuant analysis (Molecular Probes), a fluorescent dye method that measures nucleic acid content and microscopic observation.

[0288] C. Identification of C5aR inhibitors

[0289] 1. Analysis and evaluation

[0290] To evaluate small organic molecules that block ligand binding to the C5a receptor, an assay that detects binding of a radioligand (i.e., C5a) to cells expressing C5aR on the cell surface (e.g., cAMP-stimulated U937 cells or isolated human neutrophils) is used. For compounds that inhibit binding, whether competitive or not, fewer radioactivity counts are observed compared to non-inhibiting controls.

[0291] An equal number of cells are added to each well in the plate. The cells are then incubated with radiolabeled C5a. The cells are washed to remove unbound ligand, and bound ligand is determined by quantitative radioactivity counting. Cells not incubated with any organic compound provide the total count; nonspecific binding is determined by incubating cells with unlabeled ligand and labeled ligand. The percent inhibition is determined by the following formula:

[0292]

[0293] 2. Dose response curve

[0294] To determine the affinity of candidate compounds for C5aR and confirm their ability to inhibit ligand binding, 1x10 -10 Up to 1x10 -4 Inhibitory activity was determined by titration over a range of compound concentrations. In the assay, the amount of compound was varied while the cell number and ligand concentration were kept constant.

[0295] D. In Vivo Efficacy Model

[0296] By determining the efficacy of the compound in an animal model, the potential efficacy of the target compound in treating C5a-mediated diseases can be evaluated. In addition to the models described below, other suitable animal models for studying target compounds can be found in Mizuno, M et al., Expert Opin. Investig. Drugs (2005), 14(7), 807-821, which is incorporated herein by reference in its entirety.

[0297] 1. C5a-induced leukopenia model

[0298] a) C5a-induced leukopenia in a human C5aR knock-in mouse model

[0299] To study the efficacy of the compounds of the invention in animal models, recombinant mice can be created using standard techniques in which the gene sequence encoding mouse C5aR is replaced with the sequence encoding of human C5aR to create hC5aR-KI mice. In this mouse, administration of hC5a results in upregulation of adhesion molecules on the vascular wall that bind to blood leukocytes, isolating them from the bloodstream. Animals are administered 20ug / kg of hC5a, and leukocytes in peripheral blood are quantified by standard techniques after 1 minute. Pretreatment of mice with different doses of the compounds of the invention almost completely blocks hC5a-induced leukopenia.

[0300] b) C5a-induced leukopenia in cynomolgus monkey model

[0301] To investigate the efficacy of the compounds of the invention in a non-human primate model, C5a-induced leukopenia was investigated in a cynomolgus monkey model. In this model, administration of hC5a results in upregulation of adhesion molecules on the vessel wall that bind to blood leukocytes, thus sequestering them from the bloodstream. Animals were administered 10 ug / kg of hC5a, and leukocytes in peripheral blood were quantified after 1 minute.

[0302] Mouse Model of ANCA-Induced Vasculitis

[0303] On day 0, hC5aR-KI mice were injected intravenously with 50 mg / kg of purified antibody against myeloperoxidase (Xiao et al. J. Clin. Invest. 110: 955-963 (2002)). Mice were further given daily doses of the compound of the invention or vehicle orally for seven days, then the mice were sacrificed and the kidneys were collected for histological examination. Analysis of kidney sections showed a significant reduction in the number and severity of crescent-shaped and necrotic lesions in the glomeruli compared to vehicle-treated animals.

[0304] 2. Mouse Model of Choroidal Neovascularization

[0305] To investigate the efficacy of compounds of the invention in treating age-related macular degeneration (AMD), Bruch's membrane in the eyes of hC5aR-KI mice was disrupted by laser photocoagulation (Nozika et al., PNAS 103:2328-2333 (2006). Mice were treated with daily oral or appropriate intravitreal doses of vehicle or compounds of the invention for one to two weeks. Repair and neovascularization of laser-induced lesions were assessed by histology and angiography.

[0306] 3. Rheumatoid arthritis model

[0307] a) Rabbit model of destructive joint inflammation

[0308] In order to study the inhibitory effect of candidate compounds on the inflammatory response of intra-articular injection of bacterial membrane component lipopolysaccharide (LPS) in rabbits, a rabbit model of destructive joint inflammation was used. The research design mimics the destructive joint inflammation in arthritis. Intra-articular injection of LPS can cause an acute inflammatory response, which is characterized by the release of cytokines and chemokines, many of which have been identified in the joints of rheumatoid arthritis. There is a significant increase in leukocytes in synovial fluid and synovium in response to the increase in these chemotactic mediators. In this model, selective antagonists of chemokine receptors show efficacy (see Podolin et al., Journal of Immunology (J.Immunol) 169 (11): 6435-6444 (2002)).

[0309] Rabbit LPS studies were performed essentially as described by Podolin et al., supra. Female New Zealand rabbits (approximately 2 kg) were treated intra-articularly in one knee with LPS (10 ng) containing vehicle alone (phosphate buffer containing 1% DMSO) or with the addition of candidate compounds (dose 1 = 50 μM or dose 2 = 100 μM) (total volume 1.0 mL). Sixteen hours after LPS injection, the knee was lavaged and cell counts were performed. The beneficial effects of treatment were determined by histopathological assessment of synovial inflammation. Inflammation scores for histopathological assessment: 1 - minimal, 2 - mild, 3 - moderate, 4 - moderately marked.

[0310] b) Evaluation of compounds in a collagen-induced arthritis rat model

[0311] A 17-day progressive type II collagen arthritis study was conducted to assess the effect of candidate compounds on clinical ankle swelling caused by arthritis. Rat collagen arthritis is an experimental model of polyarthritis, which has been widely used in preclinical trials of many anti-arthritis drugs (see Trentham et al., J. Exp. Med. 146 (3): 857-868 (1977), Bendele et al., Toxicologic Pathology (Toxicologic Pathol.) 27: 134-142 (1999), Bendele et al., Arthritis and Rheumatism (Arthritis Rheum.) 42: 498-506 (1999)). The characteristics of this model are the occurrence and development of robust, easily measurable polyarticular inflammation, which is significantly associated with cartilage destruction and mild to moderate bone resorption and periosteal bone proliferation caused by pannus formation.

[0312] Female Lewis rats (approximately 0.2 kg) were anesthetized with isoflurane and injected with Freund's incomplete adjuvant containing 2 mg / mL bovine type II collagen at two sites, at the base of the tail and on the back, on days 0 and 6 of this 17-day study. Candidate compounds were administered subcutaneously daily at effective doses from day 0 to day 17. Caliper measurements of ankle diameter were taken, and reduced joint swelling was used as a measure of efficacy.

[0313] 4. Rat Model of Sepsis

[0314] To investigate the effects of the target compounds on inhibiting the generalized inflammatory response associated with sepsis-like illness, a cecal ligation and puncture (CLP) rat model of sepsis was used. Rat CLP studies were performed essentially as described by Fujimura N et al. (American Journal Respiratory Critical Care Medicine 2000; 161: 440-446). Briefly described herein, Wistar albino rats (both sexes) weighing between 200-250 g were fasted for 12 hours prior to the experiment. The animals were maintained on a normal 12-hour light and dark cycle and fed a standard rat chow until 12 hours prior to the experiment. The animals were then divided into four groups; (i) two sham-operated groups and (ii) two CLP groups. Each of these two groups (i.e., (i) and (ii)) was divided into a solvent control group and a test compound group. Sepsis was induced by the CLP method. Under brief anesthesia, a midline laparotomy was performed using minimal dissection, and the cecum was ligated with 3-0 silk just below the cecal valve to maintain intestinal continuity. The anti-endometrial surface of the cecum was perforated with an 18-gauge needle at two locations 1 cm apart, and the cecum was gently squeezed until feces was squeezed out. The intestine was then returned to the abdomen and the incision was closed. At the end of the operation, saline (3 ml / 100 g body weight) was injected subcutaneously to resuscitate all rats. Postoperatively, rats were fasted but had free access to water for the next 16 hours until sacrifice. The sham group underwent laparotomy and the cecum was processed but not ligated or perforated. The beneficial effects of the treatment were measured by histopathological scoring of tissues and organs and measurement of several key indicators of liver function, renal function and lipid peroxidation. To test liver function, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured. Blood urea nitrogen and creatinine concentrations were studied to assess renal function. Serum levels of proinflammatory cytokines such as TNF-α and IL-1β were also analyzed by ELISA.

[0315] 5. Mouse SLE model of experimental lupus nephritis.

[0316] To investigate the effects of target compounds on systemic lupus erythematosus (SLE), the MRL / lpr mouse SLE model was used. lpr / lpr The strain (MRL / lpr) is a commonly used mouse model of human SLE. In order to test the effectiveness of the compound in this model, male MRL / lpr mice were evenly divided into a control group and a C5aR antagonist group at 13 weeks of age. Then in the next 6 weeks, the compound or solvent (vehicle) was administered by an osmotic pump to maintain coverage and minimize the stress effect on the animal. During the six weeks of disease onset and progression, serum and urine samples were collected every two weeks. In a minority of these mice, the development of glomerular sclerosis caused the animals to die from renal failure. The post-mortem mortality rate as an indicator of renal failure is one of the criteria, and successful treatment usually leads to a delay in the onset of sudden death in the test group. In addition, the presence and magnitude of renal disease can also be continuously monitored by the measurement of blood urea nitrogen (BUN) and proteinuria. Tissues and organs were also harvested at week 19, and histopathological and immunohistochemical examinations were performed, and scores were scored according to tissue damage and cell infiltration.

[0317] 6. COPD rat model

[0318] Smoke-induced airway inflammation in rodent models can be used to evaluate the efficacy of compounds in chronic obstructive pulmonary disease (COPD). Selective antagonists of chemokines show efficacy in this model (see Stevenson et al., American Journal of Physiology-Lung Cell and Molecular Physiology) (Am.J.Physiol Lung Cell MolPhysiol.) 288L514-L522, (2005)). The acute rat model of COPD was performed as described by Stevenson et al. The compound of interest was administered systemically by oral or intravenous administration; or topically with aerosolized compounds. Male Sprague-Dawley rats (350-400g) were placed in a plexiglass chamber and exposed to cigarette smoke inhaled by a pump (50mL every 30 seconds, with fresh air in between). Rats were exposed for a total of 32 minutes. Rats were killed 7 days after the initial exposure. Any beneficial effects of treatment were evaluated by a reduction in inflammatory cell infiltration, a reduction in chemokine and cytokine levels.

[0319] In the chronic model, mice or rats are exposed to tobacco smoke every day for up to 12 months. The compound is administered orally once a day, or may be administered systemically by atomized compound topical administration. In addition to the inflammation observed in the acute model (Stevensen et al.), animals may also show other pathologies similar to human COPD, such as emphysema (indicated by an increase in the mean linear intercept) and altered lung chemistry (see Martorana et al., Am. J. Respir. Crit Care Med., 172 (7): 848-53.

[0320] 7. Mouse EAE Model of Multiple Sclerosis

[0321] Experimental autoimmune encephalomyelitis (EAE) is a model of multiple sclerosis in humans. Variants of this model have been disclosed and are well known in the art. In a typical protocol, C57BL / 6 (Charles River Laboratories) mice are used for the EAE model. On day 0, mice are subcutaneously immunized with 200ug myelin oligodendrocyte glycoprotein (MOG) 35-55 (Peptide International) emulsified in complete Freund's adjuvant (CFA) containing 4mg / ml Mycobacterium tuberculosis (Sigma-Aldrich). In addition, on days 0 and 2, animals are intravenously given 200ng pertussis toxin (Calbiochem). Clinical scoring is based on a 0-5 scale: 0, no signs of disease; 1, flaccid tail; 2, hind limb weakness; 3, hind limb paralysis; 4, forelimb weakness or paralysis; 5, moribund. Administration of the compound to be evaluated can be initiated on day 0 (prevention) or day 7 (treatment, when there is histological evidence of disease but few animals show clinical symptoms) and administered once or more daily at a concentration appropriate to their activity and pharmacokinetic properties, e.g., 100 mg / kg subcutaneously. The efficacy of the compound can be assessed by comparing severity (maximum mean clinical score in the presence of the compound compared to vehicle) or by measuring the reduction in the number of macrophages (F4 / 80 positive) isolated from the spinal cord. Spinal cord mononuclear cells can be isolated by discontinuous Percoll gradients. Cells can be stained with rat anti-mouse F4 / 80-PE or rat IgG2b-PE (Caltag Laboratories) and quantified by FACS analysis using 10 ul of Polybeads (Polysciences) per sample.

[0322] 8. Kidney transplantation mouse model

[0323] Transplantation models can be performed in mice, such as Faikah Gueler et al., JASN Express, August 27, 2008, describing an allogeneic renal transplantation model from C57BL / 6 to BALB / c mice. In short, mice are anesthetized, the left donor kidney is connected to the aortic cuff and the renal vein with a small cavity cuff, and the ureter is completely removed. After the left nephrectomy of the recipient, the vascular cuff is anastomosed with the abdominal aorta and vena cava of the recipient, respectively, at a level lower than the natural renal blood vessels. The ureter is directly anastomosed to the bladder. The cold ischemia time is 60 minutes and the warm ischemia time is 30 minutes. The right kidney can be removed at the time of allogeneic transplantation or on the 4th day after transplantation for long-term survival studies. The overall physical condition of the mice is monitored to find evidence of rejection. Compound treatment of the animals can be started immediately before surgery or after transplantation, such as subcutaneous (sub) incision injection once a day. The renal function and survival rate of mice were studied. Serum creatinine levels were measured by an automated method (Beckman Analyzer, Krefeld, Germany).

[0324] 9. Ischemia / reperfusion mouse model

[0325] Can be according to Xiufen Zheng et al., American Journal of Pathology (Am.J.Pathol), the 4th issue of volume 173, described in October 2008 ischemia / reperfusion injury mouse model.In short, CD1 mice of 6-8 weeks of age are anesthetized and placed on a heating pad to keep warm during surgery.After abdominal incision, the renal pedicle is directly dissected, and a microvascular clamp is placed on the left renal pedicle for 25-30 minutes.After ischemia, the clip is removed together with the right kidney, the incision is sutured, and the animal is allowed to recover.Blood is collected for serum creatinine and BUN analysis to indicate kidney health.Or, monitor the survival of animals over time.Compounds can be administered to animals before and / or after surgery, and the effect on serum creatinine, BUN or animal survival is used as an indicator of compound efficacy.

[0326] 10. Mouse Model of Tumor Growth

[0327] 1x105 TC-1 cells (ATCC, VA) were injected subcutaneously into the right or left posterior flank of C57BL / 6 mice aged 6-16 weeks. Starting about 2 weeks after cell injection, tumors were measured with a caliper every 2-4 days until the desired tumor size, and mice were killed. When the animals were killed, a complete autopsy was performed and the spleen and tumor were removed. The excised tumors were measured and weighed. The compound can be administered before and / or after tumor injection, and the efficacy of the compound is evaluated by delaying or inhibiting tumor growth.

[0328] Embodiment 13

[0329] The compounds in Table 1 (below) were prepared using the methods described above. Characterization data for each compound listed is provided. The activity of the chemotaxis assay using U937 cells described herein is as follows (Example 12): +, 500 nM <IC 50 ;++,50nM <IC 50 ≤500nM;+++,5nM <IC 50 ≤50nM; and ++++,IC 50 ≤5nM.

[0330] Table 1: Structure, characterization data and biological activity data of specific examples

[0331]

[0332]

[0333]

[0334]

[0335]

[0336]

[0337]

[0338]

[0339]

[0340]

[0341]

[0342]

[0343]

[0344]

[0345]

[0346]

[0347]

[0348]

[0349]

[0350]

[0351]

[0352]

[0353]

[0354]

[0355]

[0356]

[0357]

[0358]

[0359]

[0360]

[0361]

[0362]

[0363]

[0364]

[0365]

[0366]

[0367]

[0368]

[0369]

[0370]

[0371]

[0372]

[0373]

[0374]

[0375]

[0376]

[0377]

[0378]

[0379]

[0380]

[0381]

[0382]

[0383]

[0384]

[0385]

[0386]

[0387]

[0388]

[0389]

[0390]

[0391]

[0392]

[0393]

[0394]

[0395]

[0396]

[0397]

[0398]

[0399]

[0400]

[0401]

[0402] Although specific embodiments of the present invention are described herein, variations of the disclosed embodiments will become apparent to those skilled in the art after reading the specification, and it is expected that those skilled in the art may appropriately adopt such variations. Therefore, it is intended to practice the present invention in a manner different from that specifically described herein, and the present invention includes all modifications and equivalents of the subject matter recorded in the appended claims permitted by applicable law. Moreover, unless otherwise indicated herein or clearly contradictory to the context, the present invention encompasses any combination of the above elements in all possible variations thereof. All publications, patent applications, accession numbers, and other references cited in this specification are incorporated herein by reference, as if each individual publication or patent application was expressly and individually indicated to be incorporated by reference.

Claims

1. A compound of formula (I), or a pharmaceutically acceptable salt thereof, (I) in, ； X 1 Selected from the group consisting of: key, C 1-8 Alkylene, C(O), C(O)-C 1-4 Alkylene and S(O) 2 ; R 1 Select from the following group: a) a 5- to 9-membered heteroaryl group having 1 to 3 heteroatoms selected from N and S as ring vertices; b) C 6-10 Aryl; c) C 3-8 Cycloalkyl; d) a 4- to 8-membered heterocycloalkyl group having 1 O atom as a ring vertex; e) C 1-8 Alkyl, C 1-8 Alkoxy, C 1-8 Alkyl halide and –C(O)NR 1a R 1b ; Among them, R 1a and R 1b Each independently selected from the group consisting of hydrogen or C 1-8 alkyl; Among them, the group –X 1 -R 1 Optionally 1 to 5 R x Substituted by a substituent; Alternatively, the group –X 1 -R 1 Selected from and ; Each R 3 C 1-4 alkyl; R 4 Independently selected from the group consisting of: ; Each R 5 Independently selected from the group consisting of F, Cl, CN, C 1-4 Alkyl and C 1-4 Alkoxy; Each R x Independently selected from the group consisting of halogen, CN, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, hydroxy, C 2-4 Alkenyl, C 3-6 Cycloalkyl, CO 2 -C 1-4 Alkyl and CONH 2 ; subscript m is 0, 1, or 2; and Subscript n is 0, 1, or 2.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 4 Select from the following group: 。 3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 4 Select from the following group: 。 4. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that X 1 is the key.

5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that X 1 is C(O).

6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that X 1 C 1-8 Alkylene.

7. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that X 1 C(O)-C 1-4 Alkylene or S(O) 2 .

8. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 is a 5- to 9-membered heteroaryl group having 1 to 3 heteroatoms selected from N and S as ring vertices; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

9. The compound according to claim 8, It is characterized in that R 1 is selected from the group consisting of pyrazolyl, pyridinyl, pyrimidinyl, imidazolyl, thiazolyl, thiadiazolyl and pyrazinyl; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

10. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 C 6-10 Aryl; and wherein the group –X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

11. The compound according to claim 1, It is characterized in that R 1 is phenyl; and wherein the group –X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

12. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 C 3-8 Cycloalkyl; and wherein the group –X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

13. The compound according to claim 12, It is characterized in that R 1 is selected from the group consisting of cyclobutyl, cyclopentyl and cyclohexyl; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

14. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 is a 4- to 8-membered heterocycloalkyl group having 1 O atom as a ring vertex; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

15. The compound according to claim 14, It is characterized in that R 1 is selected from the group consisting of oxetanyl, tetrahydrofuranyl, and tetrahydropyranyl; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

16. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 Select from the following group: C 1-8 Alkyl, C 1-8 Alkoxy, C 1-8 Alkyl halide and –C(O)NR 1a R 1b ; Among them, R 1a and R 1b are each independently selected from the group consisting of hydrogen and C 1-8 alkyl; and wherein the group –X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

17. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 is selected from the group consisting of phenyl, pyridyl, pyrimidinyl and pyrazinyl; and wherein the group -X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent.

18. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that n is 0, 1 or 2, and if R is present 5 , then each R 5 Selected from the group consisting of F, Cl, CN, CH 3 and OCH 3 .

19. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 Selected from the group consisting of phenyl or pyridyl, wherein Group –X 1 -R 1 Optionally 1 to 4 R x Substituted by a substituent; n is 0, 1 or 2, and if R is present 5 , each R 5 Selected from the group consisting of F, Cl, CN, CH 3 and OCH 3 .

20. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 Select from the following group: 。 21. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that –X 1 -R 1 Select from the following group: 。 22. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 Select from the following group: 。 23. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 Select from the following group: 。 24. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that R 1 Select from the following group: 。 25. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that n is 0.

26. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that The compound is selected from the group consisting of: .

27. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that The compound is 。 28. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that The compound is 。 29. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that The compound is 。 30. The compound of claim 1, or a pharmaceutically acceptable salt thereof, It is characterized in that The compound is 。 31. A pharmaceutical composition comprising the compound according to any one of claims 1 to 30 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

32. The pharmaceutical composition of claim 31 , formulated for oral, intravenous, transdermal or subcutaneous administration.

33. Use of a compound as claimed in any one of claims 1 to 30 in the preparation of a medicament or formulation for treating a disease or condition selected from the group consisting of Wegener's granulomatosis, vasculitis, C3 glomerulopathy, C3 glomerulonephritis and lupus nephritis.

34. The use according to claim 33, It is characterized in that The vasculitis is microscopic polyangiitis or ANCA vasculitis.

35. The use according to claim 33, It is characterized in that The disease or condition is ANCA vasculitis.

36. The use according to claim 33, It is characterized in that The disease or condition is selected from the group consisting of Wegener's granulomatosis and microscopic polyangiitis.

37. The use according to claim 33, It is characterized in that The disease or condition is vasculitis.

38. The use according to claim 33, It is characterized in that The disease or condition is C3 glomerulopathy.

39. The use according to claim 33, It is characterized in that The disease or condition is lupus nephritis.

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