RU2780322C2 - 5-5 CONDENSED RINGS AS C5a INHIBITORS - Google Patents

Abstract

FIELD: organic chemistry.

SUBSTANCE: invention relates to the field of organic chemistry, namely to a heterocyclic compound of the formula (I) or its pharmaceutically acceptable salt, where a cycle element A¹ is selected from a group consisting of N and CH; a cycle elements A² is selected from a group consisting of N, CH, and C(R⁴); each of cycle elements A³, A⁴, A⁵, and A⁶ is independently selected from a group consisting of CH and C(R⁴); each of dotted lines is independently a simple or double bond; R¹ is selected from a group consisting of -C_1-8alkylene-heteroaryl, -C₁ alkylene-C₆ aryl, C_1-8 alkyl, -C(O)-C_1-8 alkyl, -C(O)-C₆ aryl, -C(O)-heteroaryl, -C(O)-C_3-6 cycloalkyl, -C(O)NR^1aR^1b, SO₂-C₆ aryl, -C(O)-C₁ alkylene-O-C₆ aryl, -C(O)-C_1-3 alkylene-C_3-6 cycloalkyl, and -CO₂R^1a; the heteroaryl group is a 5-6-element aromatic ring containing 1-2 heteroatoms as cycle elements, selected from N and O; where R^1a and R^1b each independently is selected from a group consisting of a hydrogen atom and C_1-8 alkyl; where R¹ is optionally substituted with 1-2 substitutes R⁵; R^2a and R^2e each is C_1-6 alkyl; R^2b, R^2c, and R^2d each is a hydrogen atom; each R³ is C_1-6 alkyl; each R⁴ is independently selected from a group consisting of C_1-6 alkyl, C_1-6 alkoxy group, C_1-6 hydroxyalkyl, halogen, cyano group, and -CO₂R^4a; each R^4a is a hydrogen atom; each R⁵ is independently selected from a group consisting of C_1-8 alkyl, C_1-8 haloalkyl, C_1-8 hydroxyalkyl, C_3-6 cycloalkyl, halogen, OH, -NR^5aR^5b, -NR^5aC(O)₂R^5b, and CO₂R^5a; where each R^5a and R^5b is independently selected from a group consisting of a hydrogen atom and C_1-8 alkyl; and subscript n is 0, 1, or 2. The invention also relates to a pharmaceutical composition based on a compound of the formula (I) or its pharmaceutically acceptable salt and to a method for the treatment of the specified diseases with a compound of the formula (I), or its salt, or a composition based on it.

EFFECT: C5a receptor modulators are obtained, useful in the treatment of diseases with pathological activation of C5a.

28 cl, 3 tbl, 13 ex

Description

The technical field to which the invention belongs

The invention relates to new low molecular weight organic modulators of the C5aR receptor, which are applicable to the suppression of pathogenic events, for example, chemotaxis associated with an increased level of anaphylatoxin activity. The invention also relates to pharmaceutical compositions and methods of use, including the treatment of diseases and disorders occurring with pathological activation of C5a, and non-pharmaceutical applications.

Link to a “sequence listing”, table or computer program listing the applications filed on the CD

Not applicable

Prior Art

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

The complement system consists of a group of proteins that are normally present in the blood plasma in an inactive state. Activation of the complement system mainly covers three different pathways, namely the classical, alternative, and lectin pathways (V. M. Holers, In Clinical Immunology: Principles and Practice, ed. R. R. Rich, Mosby Press; 1996, 363-391): 1) The classical pathway represents is a calcium/magnesium dependent cascade that is typically activated by the formation of antigen-antibody complexes. It can also be activated in an antibody-independent manner by binding to a C-reactive protein complexed with a ligand by many pathogens, including Gram-negative bacteria. 2) The alternative pathway is a magnesium-dependent cascade that is activated by the deposition and activation of C3 on certain receptive surfaces (eg, yeast and bacterial cell wall polysaccharides, and some biopolymer materials). 3) The lectin pathway involves initial binding of a mannose-dependent lectin and subsequent activation of C2 and C4, which are shared with the classical pathway (Matsushita, M. et al., J. Exp. Med. 176: 1497-1502 (1992); Suankratay, C. et al., J. Immunol 160: 3006-3013 (1998)).

Activation of the complement system generates biologically active fragments of complement proteins, eg C3a, C4a and C5a anaphylotoxins and C5b-9 membrane attack complexes (MACs), which all induce an inflammatory response by influencing leukocyte chemotaxis; activation of macrophages, neutrophils, platelets, mast cells and endothelial cells; and enhancing vascular permeability, cytolysis and tissue damage.

Complement C5a is one of the most potent pro-inflammatory mediators in the complement system. (The anaphylactic C5a peptide is 100 times more active, on a mole basis, in generating an inflammatory response than C3a.) C5a is the activated form of C5 (molecular weight 190 kDa). C5a is present in human plasma at about 80 μg/ml (Kohler, P. F. et al., J. Immunol. 99: 1211-1216 (1967)). It consists of two polypeptide chains, α and β, with approximate molecular weights of 115 kDa and 75 kDa, respectively (Tack, B. F. et al., Biochemistry 18: 1490-1497 (1979)). Biosynthesized as a single-stranded promolecule, C5 is enzymatically cleaved into a double-stranded structure during processing and secretion. After cleavage, the two resulting chains are held together by at least one disulfide bond as well as non-covalent interactions (Ooi, Y. M. et al., J. Immunol. 124: 2494-2498 (1980)).

C5 is cleaved into C5a and C5b fragments during complement activation. The convertase enzymes responsible for C5 activation are multi-subunit complexes from C4b, C2a and C3b for the classical pathway, and from (C3b) ₂ , Bb and P for the alternative pathway (Goldlust, MB et al., J. Immunol. 113 : 998-1007 (1974); Schreiber, R. D. et al, Proc. Natl. Acad. Sci. 75: 3948-3952 (1978)). C5 is activated by cleavage at position 74-75 (Arg-Leu) in the α chain. After activation, the 11.2 kDa 74-amino acid peptide C5a is released from the amino-terminal region of the α-chain. C5a and C3a are both potent neutrophil and monocyte stimulators (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)).

In addition to its anaphylotoxic properties, C5a causes chemotoxic migration of neutrophils (Ward, P. A. et al., J. Immunol. 102: 93-99 (1969)), eosinophils (Kay, A. B. et al., Immunol. 24: 969-976 (1973)), basophils (Lett-Brown, M. A. et al., J. Immunol. 117: 246-252 1976)) and monocytes (Snyderman, R. et al., Proc. Soc. Exp. Biol. Med. 138 : 387-390 1971)). C5a and C5b-9 both activate endothelial cells to produce adhesive molecules necessary for the sequestration of activated leukocytes, which are involved in the mechanisms of inflammation and tissue damage (Foreman, K. E. et al., J. Clin. Invest. 94: 1147-1155 (1994) ; Foreman, K. E. et al., Inflammation 20: 1-9 (1996); Rollins, S. A. et al., Transplantation 69: 1959-1967 (2000)). C5a also mediates inflammatory responses by inducing smooth muscle contraction, increasing vascular permeability, initiating degranulation of basophils and mast cells, 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 liver gene expression in the acute phase and enhances the overall immune response by increasing the production of TNF-α, IL-1-α, IL-6, IL-8, prostaglandins, and leukotrienes (Lambris, J. D. et al., In: The Human Complement System in Health and Disease, Volanakis, J. E. ed., Marcel Dekker, New York, pp. 83-118).

The anaphylactic and chemotactic effects of C5a are thought to work through its interaction with the C5a receptor. The human C5a receptor (C5aR) is a 52 kDa membrane-bound G protein-coupled receptor that is expressed on neutrophils, monocytes, basophils, eosinophils, hepatocytes, lung smooth muscle, and endothelial cells, as well as in tissues of the renal glomeruli (Van-Epps, D. E. et al., J. Immunol. 132: 2862-2867 (1984); Haviland, D. L. et al., J. Immunol. 154:1861-1869 (1995); Wetsel, R. A., Immunol. Leff. 44: 183- 187 (1995); Buchner, R. R. et al., J. Immunol. 155: 308-315 (1995); Chenoweth, D. E. et al., Proc. Natl. Acad. Sci. 75: 3943-3947 (1978); Zwirner , J. et al., Mol Immunol 36:877-884 (1999)). The C5aR ligand-binding site is complex and consists of at least two physically separable binding domains. One binds to the amino terminus of C5a (amino acids 1-20) and the disulfide-linked core (amino acids 21-61) and the other binds to the carboxyl terminus of C5a (amino acids 62-74) (Wetsel, R. A., Curr. Opin. Immunol. 7 : 48-53 (1995)).

C5a plays an important role in inflammation and tissue damage. In cardiopulmonary bypass and hemodialysis, C5a is generated as a result of activation of the alternative pathway of the complement system when human blood comes into contact with the artificial surface of a heart-lung machine or kidney dialysis machine (Howard, R. J. et al., Arch. Surg. 123: 1496-1501 ( 1988); Kirklin, J. K. et al., J. Cardiovasc. Surg. 86: 845-857 (1983); Craddock, P. R. et al., N. Engl. J. Med. 296: 769-774 (1977)). C5a increases capillary permeability and causes edema, bronchial constriction, pulmonary vasoconstriction, activation of leukocytes and platelets and their infiltration into tissues, in particular in the lungs (Czermak, B. J. et al., J. Leukoc. Biol. 64: 40-48 (1998) ). Administration of C5a monoclonal antibodies has been shown to reduce coronary endothelial dysfunction caused by extracorporeal circulation and cardiac arrest (Tofukuji, M. et al., J. Thorac. Cardiovasc. Surg. 116: 1060-1068 (1998)).

C5a is also implicated in acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD) and multiple organ failure (MOF) (Hack, C. E. 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 enhances monocyte production of two important pro-inflammatory cytokines, TNF-α and IL-1. C5a has also been shown to play an important role in the development of tissue damage, and in particular lung tissue, in animal models of septic shock (Smedegard G et al. Am. J. Pathol. 1989; 135: 489-497; Markus, S., et al., FASEB Journal (2001), 15: 568-570). In rat, pig, and non-human primate models of sepsis, C5a antibodies administered to animals prior to endotoxin or E. coli treatment resulted in a reduction in tissue damage as well as a decrease in 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, J. H. et al., J. Clin. Invest. 77 : 1812-1816 (1986)). More importantly, blockade of C5a by C5a polyclonal antibodies significantly improves survival in cecal ligation/puncture sepsis models in rats (Czermak, B.J. et al., Nat. Med. 5: 788-792 (1999)). This model shares many aspects with the clinical manifestations of sepsis in humans. (Parker, S. J. et al., Br. J. Surg. 88: 22-30 (2001)). In the same model of sepsis, C5a antibodies have been shown to inhibit thymocyte apoptosis (Guo, R.F. et al., J. Clin. Invest. 106: 1271-1280 (2000)) and prevent MOF (Huber-Lang, M. et al. ., J Immunol 166: 1193-1199 (2001)). C5a antibodies were also protective in the cobra venom factor rat lung injury model and in immune complex-induced lung injury (Mulligan, M. S. et al. J. Clin. Invest. 98: 503-512 (1996)). The importance of C5a in immune complex-induced lung injury was later confirmed in mice (Bozic, C. R. et al., Science 26: 1103-1109 (1996)).

C5a is an active mediator in myocardial ischemia and reperfusion. Inactivation of complement components reduced the incidence of myocardial infarction in mice (Weisman, H. F. et al., Science 249: 146-151 (1990)), and administration of C5a antibodies reduced damage in a rat model of ischemia-reperfusion in the hind legs (Bless, N. M. et al. ., Am J Physiol 276: L57-L63 (1999)). Reperfusion injury from myocardial infarction was also significantly reduced in pigs treated with monoclonal anti-C5a immunoglobulin G (Amsterdam, E. A. 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, R. D. et al., J. Thorac. Cardiovasc. Surg. 120: 350-358 (2000)).

C5a-activated neutrophils are also involved in many bullous diseases (eg, bullous pemphigoid, pemphigus vulgaris, and exfoliative pemphigus). These are chronic and recurrent inflammatory disorders clinically characterized by sterile vesicles that occur in the subepidermal space of the skin and mucosa. It is believed that antibodies to keratinocytes localized in the dermal basement membranes provide detachment of the basal keratinocytes of the epidermis from the underlying basement membrane; in this case, the bubbles are also characterized by the accumulation of neutrophils both in the higher layers of the skin and inside the cavity of the bubbles. In experimental models, a decrease in the number of neutrophils or the absence of complement components (total or C5-selective) can suppress the formation of subepidermal vesicles, even in the presence of high antibody titers.

Complement levels are elevated in patients with rheumatoid arthritis (Jose, P. J. et al., Ann. Rheum. Dis. 49: 747-752 (1990); Grant, E. P., 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, J. M. et al., Clin. Immunol. Immunopathol. 74: 283-288 (1995)). The concentration of C5a correlates with the severity of the disease state. Collagen-induced arthritis in mice and rats is similar to rheumatoid arthritis in humans. Mice deficient in C5a receptors show complete protection against arthritis induced by injection of anti-collagen monoclonal antibodies (Banda, N.K., et al., J. of Immunol., 2003, 171: 2109-2115). Therefore, suppression of C5a and/or C5a receptor (C5aR) can be used to treat these chronic diseases.

The complement system is believed to be activated in patients with inflammatory bowel disease (IBD) and is involved in the pathogenesis of this disease. Activated complement components have been found on the surface of epithelial cells as well as in the muscle layer of the mucosa and submucosal blood vessels in patients suffering from inflammatory bowel disease (IBD) (Woodruff, T.M., et al., J of Immunol., 2003, 171: 5514 -5520).

C5aR expression is upregulated in responsive astrocytes, microglia and endothelial cells in the inflamed human central nervous system (Gasque, P. et al., Am. J. Pathol. 150: 31-41 (1997)). It is possible that C5a is involved in the development of 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 C5aRs can induce apoptosis (Farkas I et al. J. Physiol. 1998; 507: 679-687). Therefore, suppression of the C5a and/or C5a receptor (C5aR) can be used to treat neurodegenerative diseases.

There is some evidence that C5a production worsens the inflammation associated with atopic dermatitis (Neuber, K., et al., Immunology 73:83-87, (1991)) and chronic allergic rash (Kaplan, A.P., J. Allergy Clin. Immunol 114, 465-474 (2004).

Psoriasis is now known to be a disease in which T cells are involved (Gottlieb, E. L. et al., Nat. Med. 1: 442-447 (1995)). However, neutrophils and mast cells may also be involved in the pathogenesis of this disease (Terui, T. et al., Exp. Dermatol. 9: 1-10; 2000); Werfel, T. et al., Arch. Dermatol. Res. 289: 83-86 (1997)). The accumulation of neutrophils under the stratum corneum of the epidermis is observed in highly inflamed areas of psoriatic plaques, and extracts from psoriatic sores contain very high concentrations of C5a and show potential chemotactic activity against neutrophils, and this effect can be suppressed by the addition of antibodies to C5a. C5a are chemoattractants for T cells and neutrophils (Nataf, S. et al., J. Immunol. 162: 4018-4023 (1999); Tsuji, R. F. et al., J. Immunol. 165: 1588-1598 (2000) ; Cavaillon, J. M. et al., Eur. J. Immunol. 20: 253-257 (1990)). In addition, C5aR expression in plasmacytoid dendritic cells (pDCs) isolated from cutaneous lupus erythematosus sores has been shown, and these cells have shown chemotactic behavior towards C5a, suggesting that blockade of C5aR on pDCs may be an effective means to reduce pDC infiltration. in inflamed skin in lupus erythematosus and psoriasis. Thus, C5a may serve as an important therapeutic target in the treatment of psoriasis.

Immunoglobulin G-containing immune complexes are implicated in the pathophysiology of a number of autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, Segren's syndrome, Goodpasture's syndrome, and hypersensitivity pneumonitis (Madaio, M. P., Semin. Nephrol. 19: 48-56 (1999); Korganow , A. S. et al., Immunity 10: 451-459 (1999); Bolten, W. K., Kidney Int. 50: 1754-1760 (1996); Ando, M. et al., Curr. Opin. Pulm. Med. 3: 391-399 (1997)). These diseases are very heterogeneous and usually affect one or more of the following organs: skin, blood vessels, joints, kidneys, heart, lungs, nervous system, and liver (including liver cirrhosis and fibrosis). The classic animal model of the inflammatory response in these immune complex diseases is the Arthus phenomenon, with polymorphonuclear cell infiltration, hemorrhage, and plasma exudation (Arthus, M., C.R. Soc. Biol. 55: 817-824 (1903)). Recent studies have shown that C5aR-deficient mice are protected from tissue damage caused by the immune complex (Kohl, J. et al., Mol. Immunol. 36: 893-903 (1999); Baumann, U. et al., J. Immunol 164: 1065-1070 (2000)). These results are in agreement with the observation that a small peptide anti-C5aR antagonist suppresses the inflammatory response caused by immune complex deposition (Strachan, A. J. et al., J. Immunol. 164: 6560-6565 (2000)). Together with its receptor, C5a plays an important role in the pathogenesis of diseases in which the immune complex is involved. C5a and C5aR inhibitors may be useful in the treatment of these diseases.

Prior Art

Non-peptidic C5a receptor antagonists have been described in the literature as being effective in the treatment of endotoxic shock in rats (Stracham, A.J., et al., J. of Immunol. (2000), 164(12): 6560-6565); and and for the treatment of IBD (inflammatory bowel disease) in a rat model (Woodruff, T.M., et al., J of Immunol., 2003, 171: 5514-5520). Non-peptide C5a receptor modulators have also been described in the patents of the Neurogen Corporation, (eg WO2004/043925, WO2004/018460, WO2005/007087, WO03/082826, WO03/08828, WO02/49993, WO03/084524); Dompe S.P.A. (WO02/029187); The University of Queenland (WO2004/100975); and ChemoCentryx (WO2010/075257).

There is strong experimental evidence in the literature that elevated C5a levels are associated with a number of diseases and disorders, in particular autoimmune and inflammatory diseases and disorders. Thus, there is a need in the art for new small molecular weight organic modulators, e.g., agonists, partial agonists, and preferably C5a receptor (C5aR) antagonists, that will be useful in suppressing pathogenic events, e.g., chemotaxis associated with increased levels of anaphylatoxin activity. The present invention satisfies this and other needs.

Brief description of the invention

In one aspect, the present invention describes compounds having formula (I):

or their pharmaceutically acceptable salt, where

cycle member A ¹ is selected from the group consisting of N, CH, C(O) and C(R ⁴ );

cycle member A ² is selected from the group consisting of N, CH and C(R ⁴ );

each of the cycle members A ³ , A ⁴ , A ⁵ and A ⁶ is independently selected from the group consisting of CH and C(R ⁴ );

each of the dotted lines independently represents a single or double bond;

R ¹ is selected from the group consisting of -C _1-8 alkylene-heteroaryl, -C _1-8 alkylene-C _6-10 aryl, C _1-8 alkyl, C _1-8 haloalkyl, -C(O)-C _{1 -8} alkyl, -C(O)-C _6-10 aryl, -C(O)-heteroaryl, -C(O)-C _3-6 cycloalkyl, -C(O)-heterocycloalkyl, -C(O)NR ^1a R ^1b , -SO ₂ -C _6-10 aryl, -SO ₂ -heteroaryl, -C (O) -C _1-8 alkylene-O-heteroaryl, -C (O) -C _1-8 alkylene-OC _{6 -10} aryl, -C(O)-C _1-8 alkylene-O- heterocycloalkyl, -C(O)-C _1-8 alkylene-O-C _3-6 cycloalkyl, -C(O)-C _1-8 alkylene-heteroaryl, -C(O)-C _1-8 alkylene-C _6-10 aryl, -C(O)-C _1-8 alkylene-heterocycloalkyl, -C(O)-C _1-8 alkylene-C _{3 -6} cycloalkyl and -CO ₂ R ^1a ; heterocycloalkyl is a 4 to 8 membered ring containing 1 to 3 heteroatoms selected from N, O and S as ring members; and the heteroaryl group is a 5 to 10 membered aromatic ring containing 1 to 3 heteroatoms selected from N, O and S as ring members;

where R ^1a and R ^1b are each independently selected from the group consisting of a hydrogen atom, C _1-8 alkyl and C _1-8 haloalkyl;

where R ¹ is optionally substituted with 1-5 substituents R ⁵ ;

R ^2a and R ^2e are each independently selected from the group consisting of C _1-6 alkyl, C _1-6 alkoxy, C _1-6 haloalkyl, -OC _1-6 haloalkyl, -SC _1-6 alkyl, -C _{1 -6} alkyl-OC _1-6 alkyl, -C _1-6 alkyl-SC _1-6 alkyl, CN and a halogen atom;

R ^2b , R ^2c and R ^2d are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 haloalkyl, -OC _1-6 haloalkyl, -SC _{1- 6} alkyl, -C _1-6 alkyl-OC _1-6 alkyl, -C _1-6 alkyl-SC _1-6 alkyl, cyano and halogen;

each R ³ is independently selected from the group consisting of C _1-6 alkyl, C _1-6 haloalkyl, halogen and hydroxyl, and optionally two R ³ groups on the same carbon atom are combined to form an oxo group (=O) or to form 3 -5-membered cycloalkyl ring;

each R ⁴ is independently selected from the group consisting of C _1-6 alkyl, C _1-6 alkoxy, C _1-6 hydroxyalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, -OC _1-6 haloalkyl , halogen, cyano, hydroxyl, -SC _1-6 alkyl, -C _1-6 alkyl-OC _1-6 alkyl, -C _1-6 alkyl-SC _1-6 alkyl, -NR ^4a R ^4b , -CONR ^4a R ^4b , -CO ₂ R ^4a , -COR ^4a , -OC(O)NR ^4a R ^4b , -NR ^4a C(O)R ^4b , -NR ^4a C(O) ₂ R ^4b and -NR ^4a -C (O)NR ^4a R ^4b ;

each R ^4a and R ^4b is independently selected from the group consisting of hydrogen, C _1-4 alkyl and C _1-4 haloalkyl;

each R ⁵ is independently selected from the group consisting of C _1-8 alkyl, C _1-8 alkoxy, C _1-8 haloalkyl, C _1-8 haloalkoxy, C _1-8 hydroxyalkyl, -C _1-8 alkylene -heterocycloalkyl, -C _1-8 alkylene-C _3-6 cycloalkyl, C _3-6 cycloalkyl, heterocycloalkyl, halogen, OH, C _2-8 alkenyl, C _2-8 alkynyl, CN, C(O)R ^5a , - NR ^5b C(O)R ^5a , -CONR ^5a R ^5b , -NR ^5a R ^5b , -C _1-8 alkylene-NR ^5a R ^5b , -SC _1-6 alkyl, -C _1-6 alkylene-OC _{1- 6} alkyl, -C _1-6 alkylene-SC _1-6 alkyl, -OC(O)NR ^5a R ^5b , -NR ^5a C(O) ₂ R ^5b , -NR ^5a -C(O)NR ^5b R ^5b and CO ₂ R ^5a ; where heterocycloalkyl group is a 4-8-membered ring containing as ring members 1 to 3 heteroatoms selected from N, O, and S;

where each R ^5a and R ^5b is independently selected from the group consisting of a hydrogen atom, C _1-8 alkyl and C _1-8 haloalkyl, or when R ^5a and R ^5b are attached to the same nitrogen atom, they are combined with a nitrogen atom with the formation of a 5 - or 6-membered ring containing 0-1 additional heteroatoms as ring members selected from N, O or S; and

subscript n is 0, 1, 2, or 3.

In addition to the compounds described herein, the present invention also describes pharmaceutical compositions containing one or more of these compounds, as well as methods for using these compounds in therapeutic methods, primarily for the treatment of diseases associated with C5a signaling activity.

In another aspect, the present invention describes methods for diagnosing a disease in a patient. In these methods, the compounds described herein are administered to a subject in labeled form, followed by diagnostic imaging to determine the presence or absence of C5aR and/or localization of cells expressing the C5aR receptor. In a related aspect, a method for diagnosing a disease is carried out by contacting a tissue or blood sample with a labeled compound as described herein and determining the presence, absence, amount, or location of C5aR in the sample.

Brief description of the drawings

Not applicable.

Detailed description of the invention

I. Abbreviations and definitions

The term "alkyl", by itself and as part of another substituent, means, unless otherwise indicated, a linear or branched hydrocarbon radical having the designated number of carbon atoms (for example, C ₁ - ₈ means 1-8 carbon atoms). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. The term "alkenyl" means an unsaturated alkyl group containing one or more double bonds. Similarly, the term "alkynyl" means an unsaturated alkyl group containing 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 their higher homologues and isomers. The term "cycloalkyl" also refers to bicyclic and polycyclic hydrocarbon rings such as, for example, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term "heterocycloalkyl" refers to a cycloalkyl group containing 1-5 heteroatoms selected from N, O, and S, where the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom(s) are optionally quaternized(s). Heterocycloalkyl may be a monocyclic, bicyclic or polycyclic ring system. Non-limiting examples of heterocycloalkyl groups 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 the like. The heterocycloalkyl group may be attached to the rest of the molecule via a ring carbon atom or a ring heteroatom.

The term "alkylene", alone or as part of another substituent, means a divalent radical derived from an alkane, exemplified by -CH ₂ CH ₂ CH ₂ CH ₂ -. Typically, an alkyl (or alkylene) group contains from 1 to 24 carbon atoms, groups having 10 or fewer carbon atoms are preferred in the present invention. "Lower alkyl" or "lower alkylene" is a short chain alkyl or alkylene group typically containing four or fewer carbon atoms. Similarly, "alkenylene" or "alkynylene" means unsaturated forms of "alkylene" containing double or triple bonds, respectively.

The term "heteroalkyl", alone or in combination with another term, means, unless otherwise indicated, a stable linear or branched chain or cyclic hydrocarbon radical, or a combination thereof, consisting of the indicated number of carbon atoms and 1-3 heteroatoms selected from the group, consisting of O, N, Si and S, where the nitrogen and sulfur atoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) O, N and S can be located in the internal position of the heteroalkyl group. The Si heteroatom can 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 -CH ₂ -CH ₂ -O-CH ₃ , -CH ₂ -CH ₂ -NH-CH ₃ , -CH ₂ -CH ₂ -N(CH ₃ )-CH ₃ , -CH ₂ -S-CH ₂ -CH ₃ , -CH ₂ -CH ₂ , -S(O)-CH ₃ , -CH ₂ -CH ₂ -S(O) ₂ -CH ₃ , -CH=CH-O-CH ₃ , -Si(CH ₃ ) ₃ , -CH ₂ -CH=N-OCH ₃ and -CH=CH-N(CH ₃ )-CH ₃ . Up to two heteroatoms can be arranged in series, as, for example, in -CH ₂ -NH-OCH ₃ and -CH ₂ -O-Si(CH ₃ ) ₃ . Similarly, the terms "heteroalkenyl" and "heteroalkynyl", alone or in combination with another term, means, unless otherwise indicated, an alkenyl group or alkynyl group, respectively, which contains the indicated number of carbon atoms and 1-3 heteroatoms selected from a group consisting of O, N, Si and S, where the nitrogen and sulfur atoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) O, N and S can be located in the internal position of the heteroalkyl group.

The term "heteroalkylene", alone or as part of another substituent, means a divalent radical, saturated or unsaturated or polyunsaturated, derived from heteroalkyl, for example -CH ₂ -CH ₂ -S-CH ₂ CH ₂ - and -CH ₂ -S-CH ₂ -CH ₂ -NH-CH ₂ -, -O-CH ₂ -CH=CH-, -CH ₂ -CH=C(H)CH ₂ -O-CH ₂ - and -S-CH ₂ -C≡C -. In the case of heteroalkylene groups, heteroatoms may also occupy one or both of the terminal positions (eg, alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like).

The terms "alkoxy", "alkylamino" and "alkylthio" (or thioalkoxy) are used in their usual sense and refer to alkyl groups attached to the rest of the molecule via an oxygen atom, an amino group or a sulfur atom, respectively. In addition, for dialkylamino groups, the alkyl moieties may be the same or different, and may also combine to form a 3-7 membered ring with the nitrogen atom to which they are attached. Accordingly, the group represented as -NR ^a R ^b includes piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl, and the like.

The term "hydroxyalkyl" is used in its usual sense and refers to a branched or linear alkyl group substituted with at least one hydroxyl group. The hydroxyl group may be at any position on the alkyl group. For example, the term "C _1-4 hydroxylalkyl" includes hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxyisopropyl, and the like.

The term "halogen" by itself or as part of another substituent means, unless otherwise indicated, a fluorine, chlorine, bromine or iodine atom. In addition, terms such as "haloalkyl" include monohaloalkyl and polyhaloalkyl. For example, the term "C _1-4 haloalkyl" includes trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term "aryl" means, unless otherwise indicated, a polyunsaturated, typically aromatic, hydrocarbon group, which may be single or multi-cycle (up to three cycles), conjugated or covalently linked. The term "heteroaryl" means aryl groups (or cycles) containing from one to five heteroatoms selected from N, O and S, where the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom(s) are optionally quaternized(s). The heteroaryl group may be attached to the rest of the molecule via a heteroatom. Non-limiting examples of aryl groups include phenyl, naphthyl, and biphenyl, and non-limiting examples of heteroaryl groups include pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, cinnolyl, phthalazinyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl, benzeneisoxazolyl, , изобензофурил, изоиндолил, индолизинил, бензотриазинил, тиенопиридинил, тиенопиримидинил, пиразолопиримидинил, пирролопиридил, имидазопиридины, бензотиаксолил, бензофуранил, бензотиенил, индолил, хинолил, изохинолил, изотиазолил, пиразолил, индазолил, птеридинил, имидазолил, триазолил, тетразолил, оксазолил, изоксазолил, тиадиазолил , pyrrolyl, thiazolyl, furyl, thienyl, and the like. The substituents in each of the above aryl or heteroaryl ring systems are selected from the group of acceptable substituents described below.

The term "pharmaceutically acceptable salts" includes salts of substances prepared with relatively non-toxic acids or bases, depending on the particular substituents in the compounds described herein. When the compounds of the present invention contain relatively acidic functionalities, base addition salts can be prepared by reacting the neutral form of such compounds with a sufficient amount of the desired base, even without a solvent or in a suitable inert solvent. Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, iron(II), iron(III), lithium, magnesium, manganese, potassium, sodium, zinc, and the like. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary and tertiary amines, including substituted amines, cyclic amines, natural amines, etc. such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, 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 prepared by reacting the neutral form of such compounds with a sufficient amount of the desired acid, without solvent or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts with inorganic acids such as hydrochloric, hydrobromic, nitric, carbonic, monohydrocarbonic, phosphoric, monohydrophosphoric, dihydrophosphoric, sulfuric, monohydrosulfuric, hydroiodic or phosphorous acid and the like, and salts with relatively non-toxic organic acids such as acetic, propionic, isobutyric, malonic, benzoic, succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts with amino acids, such as arginates and the like, and organic acid salts such as glucuronic or galacturonic acid and the like. (See for example, Berge, S. M., et al, "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Some particular compounds of the present invention contain both basic and acid functional groups, which allows such compounds to form both base addition and acid addition salts.

The neutral forms of the compounds can be regenerated by reacting the salt with a base or acid and isolating the parent compound in the usual manner. The parent form of the compound differs from the various salt forms in certain physical characteristics, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent compounds, in terms of the present invention.

In addition to salt forms, the present invention describes compounds in the form of a prodrug. Prodrugs of the compounds described in this text are compounds that easily undergo chemical changes under physiological conditions, giving the compounds of the present invention. In addition, prodrugs can be converted to the compounds of the present invention by ex vivo chemical or biological methods. For example, prodrugs can be slowly converted to the compounds of the present invention when placed in a transdermal patch with an appropriate enzyme or chemical.

Some compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to the unsolvated forms, all of which are covered by the present invention. Some compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses covered by the present invention and are within the scope of the present invention.

Some compounds of the present invention have asymmetric carbon atoms (optical centers) or double bonds; all racemates, diastereomers, geometric isomers, regioisomers, and individual isomers (eg, individual enantiomers) are within the scope of the present invention. The compounds of the present invention may also have non-natural isotope ratios of one or more of the atoms that make up these compounds. For example, compounds can be radioactively labeled with radioactive isotopes such as, for example, tritium ( ³ H), iodine-125 ( ¹²⁵ I) or carbon-14 ( ¹⁴ C). All isotopic variations of the compounds of the present invention, radioactive and non-radioactive, are within the scope of the present invention.

", которая пересекает простую, двойную или тройную связь в любой изображенной в настоящем тексте химической структуре, обозначает точку присоединения этой простой, двойной или тройной связи к остальной части молекулы.When used in real text, the wavy line"

", which crosses a single, double or triple bond in any chemical structure depicted in this text, denotes the point of attachment of this single, double or triple bond to the rest of the molecule.

I. Description of Embodiments

A. Connections

In one aspect, the present invention describes compounds having formula I:

or their pharmaceutically acceptable salt, where

cycle member A ¹ is selected from the group consisting of N, CH, C(O) and C(R ⁴ );

cycle member A ² is selected from the group consisting of N, CH and C(R ⁴ );

each of the cycle members A ³ , A ⁴ , A ⁵ and A ⁶ is independently selected from the group consisting of CH and C(R ⁴ );

each of the dotted lines independently represents a single or double bond;

R ¹ is selected from the group consisting of -C _1-8 alkylene-heteroaryl, -C _1-8 alkylene-C _6-10 aryl, C _1-8 alkyl, C _1-8 haloalkyl, -C(O)-C _{1 -8} alkyl, -C(O)-C _6-10 aryl,
-C(O)-heteroaryl, -C(O)-C _3-6 cycloalkyl, -C(O)-heterocycloalkyl, -C(O)NR ^1a R ^1b , -SO ₂ -C _6-10 aryl, -SO ₂ -heteroaryl, -C(O)-C _1-8 alkylene-O-heteroaryl, -C(O)-C _1-8 alkylene-OC _6-10 aryl, -C(O)-C _1-8 alkylene- O-heterocycloalkyl, -C(O)-C _1-8 alkylene-O-C _3-6 cycloalkyl, -C(O)-C _1-8 alkylene-heteroaryl, -C(O)-C _1-8 alkylene- C _6-10 aryl, -C(O)-C _1-8 alkylene-heterocycloalkyl, -C(O)-C _1-8 alkylene-C _3-6 cycloalkyl and -CO ₂ R ^1a ; heterocycloalkyl is a 4 to 8 membered ring containing 1 to 3 heteroatoms selected from N, O and S as ring members; and the heteroaryl group is a 5 to 10 membered aromatic ring containing 1 to 3 heteroatoms selected from N, O and S as ring members;

where R ^1a and R ^1b are each independently selected from the group consisting of a hydrogen atom, C _1-8 alkyl and C _1-8 haloalkyl;

where R ¹ is optionally substituted with 1-5 substituents R ⁵ ;

R ^2a and R ^2e are each independently selected from the group consisting of C _1-6 alkyl, C _1-6 alkoxy, C _1-6 haloalkyl, -OC _1-6 haloalkyl, -SC _1-6 alkyl, -C _{1 -6} alkyl-OC _1-6 alkyl, -C _1-6 alkyl-SC _1-6 alkyl, CN and a halogen atom;

R ^2b , R ^2c and R ^2d are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 haloalkyl, -OC _1-6 haloalkyl, -SC _{1- 6} alkyl, -C _1-6 alkyl-OC _1-6 alkyl, -C _1-6 alkyl-SC _1-6 alkyl, cyano and halogen;

each R ³ is independently selected from the group consisting of C _1-6 alkyl, C _1-6 haloalkyl, halogen and hydroxyl, and optionally two R ³ groups on the same carbon atom are combined to form an oxo group (=O) or to form 3 -5-membered cycloalkyl ring;

each R ⁴ is independently selected from the group consisting of C _1-6 alkyl, C _1-6 alkoxy, C _1-6 hydroxyalkyl, C _1-6 haloalkyl, C _1-6 haloalkoxy, -OC _1-6 haloalkyl , halogen, cyano, hydroxyl, -SC _1-6 alkyl, -C _1-6 alkyl-OC _1-6 alkyl, -C _1-6 alkyl-SC _1-6 alkyl, -NR ^4a R ^4b , -CONR ^4a R ^4b , -CO ₂ R ^4a , -COR ^4a , -OC(O)NR ^4a R ^4b , -NR ^4a C(O)R ^4b , -NR ^4a C(O) ₂ R ^4b and -NR ^4a -C (O)NR ^4a R ^4b ;

each R ^4a and R ^4b is independently selected from the group consisting of hydrogen, C _1-4 alkyl and C _1-4 haloalkyl;

each R ⁵ is independently selected from the group consisting of C _1-8 alkyl, C _1-8 alkoxy, C _1-8 haloalkyl, C _1-8 haloalkoxy, C _1-8 hydroxyalkyl, -C _1-8 alkylene -heterocycloalkyl, -C _1-8 alkylene-C _3-6 cycloalkyl, C _3-6 cycloalkyl, heterocycloalkyl, halogen, OH, C _2-8 alkenyl, C _2-8 alkynyl, CN, C(O)R ^5a , - NR ^5b C(O)R ^5a , -CONR ^5a R ^5b , -NR ^5a R ^5b , -C _1-8 alkylene-NR ^5a R ^5b , -SC _1-6 alkyl, -C _1-6 alkylene-OC _{1- 6} alkyl, -C _1-6 alkylene-SC _1-6 alkyl, -OC(O)NR ^5a R ^5b , -NR ^5a C(O) ₂ R ^5b , -NR ^5a -C(O)NR ^5b R ^5b and CO ₂ R ^5a ; where heterocycloalkyl group is a 4-8-membered ring containing as ring members 1 to 3 heteroatoms selected from N, O and S;

where each R ^5a and R ^5b is independently selected from the group consisting of a hydrogen atom, C _1-8 alkyl and C _1-8 haloalkyl, or when R ^5a and R ^5b are attached to the same nitrogen atom, they are combined with a nitrogen atom with the formation of a 5 - or 6-membered ring containing 0-1 additional heteroatoms as ring members selected from N, O or S; and

subscript n is 0, 1, 2, or 3.

With regard to the ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ and A ⁶ as cycle members, in some embodiments, the implementation of the ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ and A ⁶ as ring members is a bicyclic heteroaryl selected from

where m is 0, 1, 2 or 3; and where the R ⁴ substituents may be attached to any suitable carbon atom in the bicyclic heteroaryl.

In some embodiments, the ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ , and A ⁶ as ring members is

where m is 0, 1, 2, or 3; where the substituents R ⁴ can be attached to any suitable carbon atom in bicyclic heteroaryl.

In some embodiments, each R ⁴ is independently C _1-4 alkyl, C _1-4 alkoxy, C _1-6 hydroxyalkyl, halogen, cyano, and —CO ₂ R ^4a where R ^4a is as defined above , and where R ⁴ substituents may be attached to any suitable carbon atom in the bicyclic heteroaryl

One skilled in the art will appreciate that certain carbon atoms in the ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ and A ⁶ cannot have an R ⁴ substituent. For example, a carbon atom linking a bicyclic heteroaryl fragment (i.e., a ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ and A ⁶ ) to the rest of the molecule, and carbon atoms that are members of both ring systems in a fused bicyclic heteroaryl fragment (i.e., two carbon atoms that are members of both the benzene and the five-membered cycles) cannot have an R ⁴ substituent, because the additional substituent will exceed the valency of these carbon atoms.

In some embodiments, the ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ , and A ⁶ as cycle members is selected from the group consisting of:

In some embodiments, the ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ , and A ⁶ as cycle members is selected from the group consisting of:

With respect to R ¹ and optional substituent(s) R ⁵ , in some embodiments, R ¹ is -C _1-8 alkylene heteroaryl, -C _1-8 alkylene-C _6-10 aryl, C _1-8 alkyl, C _1-8 haloalkyl, -C(O)-C _1-8 alkyl, -C(O)-C _6-10 aryl, -C(O)-heteroaryl, -C(O)-C _3-8 cycloalkyl, -C(O)NR ^1a R ^1b , -SO ₂ -C _6-10 aryl, -C(O)-C _1-8 alkylene-OC _6-10 aryl or -CO ₂ R ^1a ; where R ^1a and R ^1b , heterocycloalkyl, and heteroaryl are as defined above, and where R ¹ is optionally substituted with 1 to 5 R ⁵ substituents.

In some embodiments, the R ¹ or R ⁵ heterocycloalkyl groups are 4-6 membered rings containing 1 to 3 heteroatoms selected from N, O, and S as ring members. In some embodiments, the R ¹ or R heteroaryl groups ⁵ are 5-6 membered aromatic rings containing 1-3 heteroatoms selected from N, O and S as ring members. In some embodiments, the C _6-10 aryl group R ¹ is phenyl.

In some embodiments, R ¹ is -CH ₂ -phenyl optionally substituted with 1-3 R ⁵ substituents.

In some embodiments, each R ⁵ is independently selected from C _1-8 alkyl, C _1-8 alkoxy, C _1-8 haloalkyl, C _1-8 haloalkoxy, C _1-8 hydroxyalkyl, C _3-6 cycloalkyl , halogen, OH,-NR ^5a R ^5b or CO ₂ R ^5a , where each R ^5a and R ^5b is independently selected from the group consisting of hydrogen, C _1-8 alkyl and C _1-8 haloalkyl.

In some embodiments, R ¹ is -CH ₂ -phenyl substituted with 1 or 2 R ⁵ substituents, where each R ⁵ is independently C _1-4 haloalkyl.

In some embodiments, R ¹ is -CH ₂ -phenyl substituted with 1 or 2 CF _{3 substituents.}

In some embodiments, R ¹ is selected from the group consisting of

In some embodiments, R ¹ is

With respect to formula I and substituents R ^2a , R ^2b , R ^2c , R ^2d and R ^2e , in some embodiments, R ^2a and R ^2e are each independently C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 an alkoxy group, -OC _1-6 haloalkyl or halogen. In some embodiments, R ^2b , R ^2c and R ^2d are independently H, C _1-4 alkyl, C _1-4 haloalkyl, or halo.

In some embodiments, R ^2b , R ^2c and R ^2d are each H.

In some embodiments, R ^2a and R ^2e are each independently C _1-6 alkyl or C _1-6 haloalkyl.

In some embodiments, R ^2a and R ^2e are each independently methyl or ethyl. In some embodiments, R ^2a and R ^2e are both methyl or both are ethyl.

In some embodiments, the fragment of formula I below

представляет собой

represents

или

.

or

.

Each R ³ in formula I, in some embodiments, is independently C _1-6 alkyl, C _1-6 haloalkyl, or halogen. In some embodiments, each R ³ is independently C _1-4 alkyl.

In some embodiments, n, the subscript in R ³ is 0, 1, or 2. In some embodiments, n is 0. In some embodiments, n is 2.

In some embodiments, the moiety of Formula I represented by

представляет собой

,

, или

.

represents

,

, or

.

In some embodiments, the compound having formula I is represented by formula (Ia), (Ib) or (Ic).

In embodiments in which a compound having formula (I) has formula (Ia), R ¹ , R ³ , n, R ^2a , R ^2e , and a ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ , and A ⁶ as members of the cycle correspond to the above definitions for formula (I).

In embodiments in which a compound having formula (I) has formula (Ib), R ¹ , R ^2a , R ^2e , and a ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ and A ⁶ as cycle members correspond to the above definitions for formula (I).

In embodiments in which a compound having formula (I) has formula (Ic), R ¹ and a ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ and A ⁶ as ring members correspond to the above definitions for formula (I).

In some embodiments, a compound having formula (I) has formula (Id), (Ie), or (If)

In embodiments in which a compound having formula (I) has formula (Id), R ¹ , R ⁴ , m, R ^2a and R ^2e are as defined above for formula (I).

In embodiments in which a compound having formula (I) has formula (Ie), R ¹ , R ⁴ and m are as defined above for formula (I).

In embodiments in which a compound having formula (I) is of formula (If), R ⁵ , R ⁴ , and m are as defined above for formula (I), and p is 0, 1, or 2.

In some embodiments, a compound having formula (I) has formula (Ig) or (Ih)

In embodiments in which a compound having formula (I) has formula (Ig) or (Ih), R ⁴ and m are as defined above for formula (I).

In some embodiments, the compound having formula (I) is the compound described in the Examples section.

Getting connections

Some compounds of the present invention can be obtained according to the methods described in this application in the Examples section. In addition, syntheses of certain intermediates that can be used to synthesize the compounds of the present invention are also described.

B. Pharmaceutical compositions

In addition to the compounds described above, compositions for modulating C5a activity in humans and animals typically contain a pharmaceutically acceptable carrier or diluent.

The term "composition" as used in this text includes a product containing the specified ingredients in the specified amounts, as well as any product that is obtained, directly or indirectly, by combining the specified ingredients in the specified amounts. The term "pharmaceutically acceptable" means a carrier, diluent or excipient that must be compatible with the other ingredients of the drug and not cause harm to the patient receiving it.

Pharmaceutical compositions for administering the compounds of the present invention may be presented in unit dosage form and may be prepared by any of the methods known in the art of pharmacology and drug administration. All methods include the step of bringing into association the active substance with the carrier which contains one or more accessory ingredients. In general, pharmaceutical compositions are prepared by uniformly and intimately mixing the active ingredient with a liquid carrier or a finely divided solid carrier, or both, and then, if necessary, shaping the product into the desired formulation. In the pharmaceutical composition, the active substance is present in an amount sufficient to provide the desired effect on the disease or pathological condition.

Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral administration, for example, they may be in the form of tablets, sachets, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions and self-emulsifying preparations, as described in the Patent Application. US No. 6,451,339, hard or soft capsules, syrups, elixirs, solutions, buccal patches, oral gel, chewing gum, chewable tablets, effervescent powders and effervescent tablets. Compositions intended for oral administration may be prepared by any means known in the pharmaceutical art. Such compositions may contain one or more agents selected from the group consisting of sweeteners, flavors, colors, antioxidants, and preservatives to provide pharmaceutically attractive and palatable formulations. The tablets contain the active ingredient mixed with non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets. Such excipients may be, for example, inert diluents such as cellulose, carbon dioxide, alumina, calcium carbonate, sodium carbonate, glucose, mannitol, sorbitol, lactose, calcium phosphate or sodium phosphate; granulating agents and agents that accelerate the disintegration of tablets, such as corn starch or alginic acid; binding agents such as polyvinylpyrrolidone, cellulose, starch, gelatin or acacia gum; and lubricants such as magnesium stearate, stearic acid or talc. Tablets may be uncoated, or they may be enteric-coated, or coated in some other known manner to slow disintegration and absorption in the gastrointestinal tract, thereby providing prolonged action for a longer time. For example, disintegrating materials such as glyceryl monostearate or glyceryl distearate can be used. Tablets can also be coated by the methods described in US Pat. Nos. 4,256,108; 4,166,452 and 4,265,874 to form sustained release osmotic tablets.

Preparations intended for oral administration may also be in the form of hard gelatin capsules, where the active substance is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin, or in the form of soft gelatin capsules, where the active substance is mixed with an aqueous or oily medium, for example, peanut butter, liquid paraffin or olive oil. In addition, it is possible to prepare emulsions with water-insoluble ingredients such as oils and stabilize them with surfactants such as mono- or diglycerides, PEG esters, and the like.

Aqueous suspensions contain the active ingredients mixed with excipients suitable for the production of aqueous suspensions. Such excipients are suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and acacia gum; dispersing or wetting agents can be natural phosphatides, such as lecithin, or alkylene oxide condensates with fatty acids, eg polyoxyethylene stearate, or ethylene oxide condensates with long chain aliphatic alcohols, eg heptadecaethyleneoxycetanol, or ethylene oxide condensates with partial esters derived from fatty acids and hexitol, such as polyoxyethylene sorbitol monooleate, or the condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl or more colors, one or more flavors, and one or more sweeteners such as sucrose or saccharin.

Oil suspensions can be prepared by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. Oily suspensions may contain thickeners such as beeswax, hard paraffin or cetyl alcohols. Sweetening agents such as those listed above and coloring agents may be added to provide a palatable oral composition. Such compositions can be stabilized by the addition of an antioxidant such as ascorbic acid.

Dispersible powders and granules suitable for the preparation of an aqueous suspension with the addition of water contain the active ingredient in admixture with a dispersing or wetting agent, a suspending agent and one or more preservatives. Examples of dispersing or wetting agents, suspending agents are given above. Additional adjuvants may also be present, such as sweeteners, flavors and colors.

The pharmaceutical compositions of the present invention may also be in the form of oil-in-water emulsions. The oil phase can be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin, or a mixture thereof. Suitable emulsifiers can be natural gums such as acacia gum or gum tragacanth, natural phosphatides such as soybean oil, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of said partial esters. ethers with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners and flavors.

Syrups and elixirs may contain sweeteners such as glycerin, propylene glycol, sorbitol or sucrose. Such formulations may also contain emollients, a preservative, flavorings and coloring agents. Oral solutions can be prepared in combination with, for example, cyclodextrin, PEG and surfactants.

Pharmaceutical compositions may take the form of sterile injectable aqueous or oily suspensions. Such a suspension can be prepared according to methods known in the art using suitable dispersants or wetting agents and suspending agents as mentioned above. Sterile injectable preparations may also be sterile injectable solutions or suspensions in a non-toxic parenterally acceptable diluent or solvent, for example, may be in the form of a solution in 1,3-butanediol. Suitable vehicles and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile oils are often used as a solvent or suspending medium. Any bland fixed oil can be used for this purpose, including synthetic mono- and diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The compounds described herein may also be administered in the form of suppositories for rectal administration of the drug. Such compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at room temperature but liquefies at body temperature and therefore melts in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycols. In addition, the compounds can be administered as ophthalmic preparations as drops or ointments. It is also possible to carry out transdermal administration of the subject compounds via iontophoresis patches and the like. For topical application, creams, ointments, gels, solutions or suspensions containing the compounds of the present invention are used. In the context of the present invention, topical application also includes the use of rinses and mouth solutions.

The compounds of the present invention may also be combined with suitable polymer carriers as targeted drug carriers. Such polymers may include polyvinylpyrrolidone, a pyran copolymer, polyhydroxypropyl methacrylamide phenol, polyhydroxyethyl aspartamide phenol or polyethylene oxide polylysine substituted with palmitoyl residues. In addition, the compounds of the present invention can be combined with a carrier belonging to the class of biodegradable polymers that can be used to achieve controlled drug release, for example: polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, poly-epsilon-caprolactone, polyhydroxybutyric acid , polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and cross-linked or amphipathic hydrogel block copolymers. Polymers and semi-permeable polymer matrices can be molded into articles such as valves, stents, tubes, prostheses, and the like. In one embodiment of the present invention, the compound of the present invention is combined with a polymer or a semi-permeable polymeric matrix formed into a stent or stent graft.

Pharmaceutical compositions of the present invention may contain one or more additional therapeutic agents. These one or more additional therapeutic agents may include corticosteroids, steroids, immunosuppressants, or CD 20 inhibitors. In some embodiments, one or more additional therapeutic agents include obinutuzumab, rituximab, ocrelizumab, cyclophosphamide, prednisolone, hydrocortisone, hydrocortisone acetate, cortisone acetate, thixocortol pivalate, prednisolone, methylprednisolone, triamcinolone acetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide , fluocinonide, fluocinolone acetonide, halcinonide, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortolone, hydrocortisone-17-valerate, halomethasone, aclomethasone dipropionate, beclomethasone, betamethasone valerate, betamethasone dipropionate, prednicarbate, ketalobizonate-1 -17-propionate, fluocortolone caproate, fluocortolone pivalate, flupredniden acetate, hydrocortisone-17-butyrate, hydrocortisone-17-aceponate, hydrocortisone-17-buteprate, ciclesonide and prednicarbate. Additional discussion of combination therapy is included in the present application in the "Methods of Use" section.

C. Methods of application

The compounds of the present invention can be used as agonists, (preferably) antagonists, partial agonists, inverse agonists of C5a receptors in various contexts, both in vitro and in vivo. In one embodiment, the compounds of the present invention are a C5aR antagonist that can be used to inhibit the binding of a C5a receptor ligand (eg, C5a) to the C5a receptor in vitro or in vivo. In general, such methods include the step of contacting the C5a receptor with a sufficient amount of one or more of the C5a receptor modulators described herein in the presence of a C5a receptor ligand in aqueous solution and under conditions suitable to bind that ligand to the C5a receptor. The C5a receptor may be present in suspension (eg, as an isolated membrane or cell preparation), in a grown or isolated cell, or in a tissue or organ.

Preferably, the amount of C5a receptor modulator contacting the receptor should be sufficient to inhibit C5a binding to the C5a receptor in vitro and is measured, for example, using a radioligand assay, a calcium mobilization assay, or a chemotaxis assay as described herein.

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

When the compounds of the present invention are used to inhibit C5a receptor-mediated cellular chemotaxis, preferably leukocyte (e.g., neutrophil) chemotaxis, in an in vitro chemotaxis assay, such methods include contacting white blood cells (particularly primate white blood cells, especially white blood cells). human bodies) with one or more compounds of the present invention. Preferably, the concentration is sufficient to inhibit white blood cell chemotaxis in an in vitro chemotaxis assay such that the level of chemotaxis observed in the control is significantly higher than that observed in the assay supplemented with a compound of the present invention.

In another embodiment, the compounds of the present invention may also be used to treat patients suffering from pathological conditions sensitive to C5a receptor modulation. As used herein, the term "treatment" encompasses both disease-modifying treatment and symptomatic treatment, either of which may be prophylactic (i.e., before the onset of symptoms, to prevent, delay, or lessen the severity of symptoms) or therapeutic ( i.e. after the onset of symptoms, to reduce the severity and/or duration of symptoms). As used herein, a condition is considered "responsive to C5a receptor modulation" if modulation of C5a receptor activity results in a decrease in abnormal C5a receptor activity. As used herein, the term "patients" includes primates (especially humans), companion animals (such as dogs, cats, horses, and the like), and farm animals (such as cattle, pigs, sheep, and etc.), where the dosages are as described in this text.

Conditions that can be treated with C5a modulation:

Autoimmune diseases - eg, 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, hard sediment disease, pemphigus, pemphigoid, scleroderma, myasthenia gravis, autoimmune hemolytic and thrombocytopenic conditions, Goodpasture's syndrome (and associated glomerulonephritis and pulmonary hemorrhage), C3 glomerulopathy, C3 glomerulonephritis, mesangioproliferative glomerulonephritis, Kawasaki disease, IH nephropathy, immunovasculitis, tissue graft rejection, "graft versus host", hyperacute rejection of transplanted organs; etc.

Inflammatory disorders and related pathological conditions - for example, neutropenia, sepsis, septic shock, Alzheimer's disease, multiple sclerosis, neutrophilia, stroke, inflammatory bowel disease (IBD), inflammation due to severe burns, lung injury and ischemia-reperfusion injury, osteoarthritis, and adult acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), systemic inflammatory response syndrome (SIRS), atopic dermatitis, psoriasis, chronic urticaria and multiple organ failure syndrome (MODS), hemolytic uremic syndrome, atypical hemolytic uremic syndrome (aHUS). Also included are pathological complications associated with insulin-dependent diabetes mellitus (including diabetic retinopathy), lupus nephropathy, Hayman's nephritis, membranous nephritis and other forms of glomerulonephritis, contact sensitive response, and inflammation due to contact of blood with artificial surfaces, which can cause complement activation, for example, during extracorporeal circulation (for example, during hemodialysis or through a cardiopulmonary resuscitator, for example, in connection with vascular surgery such as coronary artery bypass surgery or heart valve replacement), or in connection with contact with the surface of other artificial vessels or containers (e.g., ventricular support device, artificial heart machine, transfusion tubes, blood storage bags, plasmapheresis, thrombophoresis, etc.) Also included are diseases related to ischemia-reperfusion injury, such as those arising from graft transplantation, including solid organ transplantation, and syndromes such as ischemic-reperfusion injury, ischemic colitis, and cardiac ischemia. The compounds of the present invention may also be used in the treatment of age-related macular degeneration (Hageman et al, P.N.A.S. 102: 7227-7232, 2005).

Cardiovascular and cerebrovascular disorders - for example, myocardial infarction, coronary artery thrombosis, vascular occlusion, postoperative vascular reocclusion, atherosclerosis, traumatic injury to the central nervous system, and coronary heart disease. In one embodiment, an effective amount of a compound of the present invention may be administered to a patient who is at risk of developing myocardial infarction or thrombosis (e.g., who has one or more recognized risk factors for myocardial infarction or thrombosis, such as (but not limited to) ) obesity, smoking, high blood pressure, hypercholesterolemia, earlier or hereditary cases of myocardial infarction or thrombosis to reduce the risk of myocardial infarction or thrombosis.

Cancers or disorders—eg, melanoma, lung cancer, lymphoma, sarcoma, carcinoma, fibrosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, 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, embryonic carcinoma, Wilm's tumor, pleomorphic adenoma, liver cell papilloma, renal tubular adenoma, cystadenoma, papilloma, adenoma , rhabdomyoma, hemangioma, lymphangioma, osteoma, chondroma, lipoma and fibroma.

Vasculitic diseases - Vasculitic diseases are characterized by inflammation of blood vessels. Leukocyte infiltration leads to destruction of the vascular wall, and complement activity is believed to play an important role in initiating leukocyte migration as well as resulting damage at the site of inflammation (Vasculitis, Second Edition, Edited by Ball and Bridges, Oxford University Press, pp 47-53 , 2008). The compounds described in this application can be used to treat leukoclastic vasculitis, ANCA-associated vasculitis (vasculitis in which antineutrophil cytoplasmic antibodies are detected in the blood), immune vasculitis, Wegener's granulomatosis, microscopic polyangiitis, Churg-Strauss syndrome, Schonlein-Henoch purpura, nodular periarteritis , rapidly progressive glomerulonephritis (RPGN), cryoglobulinemia, giant cell arteritis (GCA), Behçet's disease, and aortic arch syndrome.

HIV Infection and AIDS - The C5a receptor modulators described herein can be used to suppress HIV infection, slow the progression of AIDS, or reduce the severity of symptoms of HIV infection and AIDS.

Neurodegenerative Disorders and Related Diseases - In other aspects, the C5a antagonists described herein can be used to treat Alzheimer's disease, multiple sclerosis, and cognitive decline associated with cardiopulmonary bypass surgery and similar procedures.

In one embodiment of the present invention, the compounds of the present invention can be used to treat diseases selected from the group consisting of sepsis (and related disorders), COPD, rheumatoid arthritis, lupus nephritis, and multiple sclerosis.

The methods of treatment described in the present invention include the introduction to the patient of an effective amount of one or more of the compounds described in this text. Suitable patients include patients suffering from a disease or disorder or undergoing (ie, prophylactic treatment) a disease or disorder referred to herein. Typical patients for the treatment described in the present invention include mammals, in particular primates, especially humans. Other suitable patients include companion animals such as dogs, cats, horses, and the like, or farm animals such as cattle, pigs, sheep, and the like.

In general, the methods of treatment described herein comprise administering to a patient an effective amount of one or more of the compounds described herein. In a preferred embodiment, the compound(s) of the present invention is preferably administered to a patient (eg, human) orally or topically. An effective amount may be an amount sufficient to modulate C5a receptor activity and/or an amount sufficient to reduce or alleviate symptoms in a patient. Preferably, the amount administered is sufficient to create a plasma concentration of the compound (or its active metabolite if the compound is a prodrug) high enough to detectably inhibit white blood cell (eg, neutrophil) chemotaxis in vitro. Treatment regimens may vary depending on the compound used and the particular condition being treated; for the treatment of most disorders, the preferred frequency of administration is 4 times a day or less. In general, twice daily administration is more preferred, and once daily administration is particularly preferred. It should be understood, however, that the specific dosage level and mode of administration for any particular patient depends on a number of factors, including the potency of the particular compound, age, body weight, general health, sex, diet, time of administration, route of administration, rate of elimination, combination of drugs. (for example, other drugs that are administered to the patient) and the severity of the specific disease being treated, as well as the opinion of the attending physician. In general, it is preferable to use the lowest dosage sufficient to provide effective therapy. Therapeutic efficacy for patients may be monitored by medical or veterinary criteria appropriate to the particular condition being treated.

Dosages on the order of about 0.1 mg to about 140 mg per kilogram of body weight per day may be used in the treatment or prevention of conditions involving pathogenic C5a activity (from about 0.5 mg to about 7 g per person per day). The amount of active ingredient that can be combined with carriers to produce a single dosage form varies depending on the particular patient being treated and on the method of administration used. Dosage formulations typically contain from about 1 mg to about 500 mg of the active ingredient. For compounds that are administered orally, transdermally, intravenously or subcutaneously, it is preferred that a sufficient amount of the compound be administered to achieve a blood plasma concentration of 5 ng (nanogram)/mL to 10 μg (microgram)/mL blood plasma, more preferably a sufficient amount compounds to achieve plasma concentrations of 20 ng/mL to 1 μg/mL plasma, most preferably a sufficient amount of the compound to achieve plasma concentrations of 50 ng/mL to 200 ng/mL plasma. For direct administration to the synovium (for the treatment of arthritis), a sufficient amount of the compound should be administered to locally achieve about 1 micromolar concentration.

The frequency of administration may also vary depending on the compound used and the particular condition being treated. However, for the treatment of most diseases, an administration regimen of 4 times a day, three times a day or less is preferred, with one administration regimen or 2 times a day being particularly preferred. It should be understood, however, that the specific dosage level for any given patient will depend on a number of factors, including the activity of the particular compound, age, body weight, general health, sex, diet, time of administration, route of administration, rate of elimination, drug combination (e.g., other drugs that are administered to the patient), the severity of the specific disease being treated, and other factors, including the opinion of the attending physician.

Combination Therapy

The compounds described herein may be used in combination with one or more additional therapeutic agents that are used in the treatment, prevention, suppression or alleviation of diseases or conditions for which the compounds and compositions of the present invention may be used. Such one or more additional therapeutic agents can be administered in the manner and in the amounts normally used for them, simultaneously or sequentially with the compound or composition of the present invention. When a compound or composition of the present invention is used simultaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the compound or composition of the present invention is preferred. Accordingly, the pharmaceutical compositions of the present invention include those which also contain one or more other active substances or therapeutic agents, in addition to the compound or composition of the present invention.

Examples of one or more additional therapeutic agents that can be combined with a compound or composition of the present invention, whether administered alone or as part of the same pharmaceutical compositions, include (but are not limited to): (a) VLA-4 antagonists, (b) steroids and corticosteroids such as beclomethasone, betamethasone (including betamethasone sodium phosphate, betamethasone valerate, betamethasone dipropionate), prednisolone, prednisolone, methylprednisolone, mometasone, dexamethasone (including dexamethasone sodium phosphate), fluticasone, cortisone (including cortisone acetate), hydrocortisone (including hydrocortisone acetate, hydrocortisone-17-valerate, hydrocortisone-17-butyrate, hydrocortisone-17-aceponate, hydrocortisone-17-buteprate), butesonide, desonide, fluocinonide (including fluocinonide acetonide), triamcinolone (including triamcinolone acetonide and triamcinolone alcohol), thixocortol (including thixocortol pivalate), fluocortolone (including fluocortolone caproate and fluocortolone pivalate ), amcinonide, halcinonide, halomethasone, fluprednidene acetate, salmeterol, salbutamol, ciclesonide, formaterol, alklomethasone (including alklomethasone dipropionate), prednicarbate, clobetasone (including clobetasone-17-butyrate), clobetasol (including clobetasol-17-propionate); (c) immunosuppressants such as cyclosporine (cyclosporin A, Sandimmune®, Neoral®), tacrolimus (FK-506, Prograf®), rapamycin (sirolimus, Rapamune®) and other FK-506 type immunosuppressants, and mycophenolate, e.g. mycophenolate mofetil (CellCept8); (d) antihistamines (H1-histamine antagonists) such as brompheniramine, chlorpheniramine, dexchlorpheniramine, triprolidine, clemastine, diphenhydramine, diphenylpyralin, tripelennamine, hydroxyzine, methdilazine, promethazine, trimeprazine, azatadine, cycloheptadiene, antazoline, pheniramine, pyrilamine, astemizole, loratadine, cetirizine, fexofenadine, descarboethoxyloratidine, etc.; (e) non-steroidal anti-asthma drugs (eg, terbutaline, metaproterenol, fenoterol, isoetharine, albuterol, bitolterol, and pirbuterol), theophylline, sodium cromolyn, atropine, ipratorium bromide, leukotriene receptor antagonists (eg, zafirlukast, montelukast, pranlukast, iralukast, pobilukast, and SKB-106,203), leukotriene biosynthesis inhibitors (Cileuton, BAY-1005); (f) non-steroidal anti-inflammatory drugs (NSAIDs) such as propionic acid derivatives (e.g. alminoprofen, benoxaprofen, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pyrprofen, pranoprofen , suprofen, thiaprofenic acid, and thioxaprofen), acetic acid derivatives (e.g., indomethacin, acemethacin, aclofenac, clidanac, diclofenac, fenclofenac, fenclosic acid, fentiazac, furofenac, ibufenac, isoxepac, oxpinac, sulindac, thiopinac, tolmetin, zidomethacin, and zomepirac) , fenamic acid derivatives (eg, flufenamic acid, meclofenamic acid, mefenamic acid, niflumic acid, and tolfenamic acid), biphenylcarboxylic acid derivatives (eg, diflunisal and flufenisal), oxicams (eg, isoxicam, piroxicam, sudoxicam, and tenoxicam), salicylates (eg. , acetylsalicylic acid, and sulfasalazine) and pyrazolones (eg, apazon, bezpi perilone, feprazone, mofebutazone, oxyphenbutazone and phenylbutazone); (g) cyclooxygenase-2 (COX-2) inhibitors such as celecoxib (Celebrex®) and rofecoxib (Vioxx®); (h) phosphodiesterase type IV (PDE IV) inhibitors; (i) gold compounds such as auranofin and aurothioglucose, (j) etanercept (Enbre10), (k) cyclophosphamide, (l) antibody therapy such as orthoclone (OKT3), daclizumab (Zenapax®), basiliximab (Simulect®), and infliximab (Remicade®), (m) CD20-targeting antibody therapy such as obinutuzumab, rituximab, or ocrelizumab; (n) chemotherapy agents such as anthracyclines (eg daunorubicin (daunomycin; rubidomycin), doxorubicin, epirubicin, idarubicin and valrubicin), mitoxantrone and pixantrone; platinum-containing agents (eg, cisplatin, carboplatin, oxaliplatin, satraplatin, picoplatin, nedaplatin, triplatin, and lipoplatin); tamoxifen and its metabolites such as 4-hydroxy tamoxifen (afimoxifen) and N-desmethyl-4-hydroxy tamoxifen (endoxifen); taxanes such as paclitaxel (taxol) and docetaxel; alkylating agents (eg, nitrogen mustards such as mechlorethamine (HN2), cyclophosphamide, ifosfamide, melphalan (L-sarcolysine) and chlorambucil); ethyleneimines and methylmelamines (eg, hexamethylmelamine, thiotepa, alkylsulfonates such as busulfan, nitrosoureas such as carmustine (BCNU), lomustine (CCNLJ), semustine (methyl-CCN-U) and streptozoin (streptozotocin), and triazenes such as decarbazine (DTIC; dimethyltriazenoimidazolecarboxamide)); antimetabolites (eg, folate analogs such as methotrexate (ametopterin), pyrimidine analogs such as fluorouracil (5-fluorouracil; 5-FU), floxuridine (fluorodeoxyuridine; FUdR), and cytarabine (cytosine arabinoside), and purine analogs and related inhibitors , such as mercaptopurine (6-mercaptopurine; 6-MP), thioguanine (6-thioguanine; 6-TG), and pentostatin (2'-deoxycophonnicin)); (o) other chemokine receptor antagonists, especially CXCR2, CXCR3, CCR2, CCR3, CCR4, CCR7, CX3CR1 and CXCR6.

The target disease or disorder determines which additional therapeutic agent or therapeutic agents are most suitable for administration in combination with the compounds of the present invention - this can be determined by a person skilled in the art.

The weight ratio of the compound of the present invention to the second active ingredient may vary and depends on the effective dosage of each ingredient. Generally, an effective dosage of each ingredient is used. Thus, for example, when a compound of the present invention is combined with an NSAID, the weight ratio of the compound of the present invention to the NSAID is typically in the range of about 1000:1 to about 1:1000, preferably about 200:1 to about 1:200. Combinations of a compound of the present invention and other active ingredients will usually also fall within the indicated ranges, but in each case the effective dosage of each active ingredient should be used.

Non-pharmaceutical applications

In another aspect of the present invention, the compounds of the present invention can be used in various non-pharmaceutical in vitro and in vivo fields. For example, the compounds of the present invention can be labeled and used as a probe for detection and localization of the C5a receptor (cell preparations or tissue section samples). The compounds of the present invention can also be used as positive controls in C5a receptor activity assays, i.e. as standards for determining the ability of a candidate to bind to the C5a receptor, or as radioactively labeled tracers for positron emission tomography (PET) or single photon emission computed tomography (SPECT). Such methods can be used to characterize C5a receptors in living subjects. For example, a C5a receptor modulator can be labeled using a number of well-known techniques (eg, administering a radionuclide label such as tritium) and incubated with the sample for the appropriate incubation time (eg, first determined by tracking binding dynamics). After incubation, the unbound compound is removed (e.g., by washing) and the bound compound is detected by any method suitable for the label used (e.g. radioautography or scintillation activity measurement for radioactively labeled compounds; spectroscopic methods can be used to detect luminescent groups and fluorescent groups ). For a control, a corresponding sample containing a labeled compound and a larger (eg, 10 times larger) amount of an unlabeled compound can be tested in a similar manner. A greater amount of detectable label remaining in the test sample than in the control sample indicates the presence of the C5a receptor in the sample. Detection including receptor autoradiography (receptor mapping) for the C5a receptor in cell cultures or tissue samples can be performed as described by Kuhar in sections 8.1.1 to 8.1.9 in Current Protocols in Pharmacology (1998) John Wiley & Sons, New York.

The compounds described herein may also be used in various well known cell separation techniques. For example, modulators can be coupled to the inner surface of a cell growth plate or other support, for use as affinity ligands to immobilize and consequently isolate C5a receptors (eg, to isolate receptor-expressing cells) in vitro. In one preferred application, a modulator associated with a fluorescent marker, such as fluorescein, is contacted with cells, which are then analyzed (or isolated) by fluorescent activated cell sorting (FACS).

II. Examples

The following examples are given to illustrate and not to limit the scope of the present invention.

The reagents and solvents described below can be obtained from commercial sources such as Aldrich Chemical Co. (Milwaukee, Wisconsin, USA). ¹ H-NMR spectra were recorded on a Varian Mercury 400 MHz NMR spectrometer. Chemical shift values are given relative to tetramethylsilane (TMS) and are described in order of multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet) and number of protons. Mass spectrometry results are given as mass divided by charge followed by the relative intensity of each ion (in brackets). In the examples, one m/e value is given for each M+H (or MH if indicated) of an ion containing the most common atomic isotopes. In all cases, the isotopic distribution corresponds to the expected formula. Electrospray ionization (ESI) mass spectrometry was performed on a Hewlett-Packard MSD electrospray ionization mass spectrometer equipped with an HP1100 HPLC for sample injection. Typically, the analyte was dissolved in methanol at a concentration of 0.1 mg/ml, and 1 microliter of the resulting solution was injected into a mass spectrometer scanning ions in the range from 100 to 1500 daltons. All compounds were analyzed in positive ion ESI mode using acetonitrile/water with 1% formic acid as solvent. The compounds described below can be analyzed in ESI mode with negative ion detection using 2 mM NH ₄ OAc in acetonitrile/water as solvent.

In the Examples and in the rest of the text of the application, the following abbreviations apply:

EtOH: Ethanol

EtONa: Sodium ethoxide

THF: Tetrahydrofuran

TLC: Thin layer chromatography

MeOH: Methanol

The compounds of the present invention can be synthesized as described below using a variety of reactions known to those skilled in the art. One of skill in the art will also appreciate that alternative methods for synthesizing the target compounds may be used, and that the approaches used herein are not exhaustive but provide workable and practical methods for preparing the compounds of the present invention.

Some of the molecules claimed in this patent may exist in various enantiomeric and diastereomeric forms, and all such compound variants are within the scope of the present invention.

The detailed description of the experimental procedures used to synthesize the key compounds in the present text lead to molecules that are described by physical data that identifies them, as well as chemical structure images that describe them.

Those skilled in the art will also appreciate that acids and bases are often used during standard processing procedures in organic chemistry. Salts of the parent compounds are sometimes formed, if they have the required intrinsic acidity or basicity, during the experimental procedures described in this text.

Example 1

Synthesis of the intermediate 6-fluoro-7-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole

Step a: A solution of vinylmagnesium bromide in THF (1.0 M, 70 ml, 70 mmol) was added to a solution of 4-bromo-2-fluoro-6-nitroanisole (5.0 g, 20 mmol) in anhydrous THF (70 ml) under nitrogen at - 50°C. The mixture was stirred at the same temperature and left to warm to -30°C for 1.5 hours. The reaction was quenched with saturated aqueous NH ₄ Cl and allowed to warm to room temperature over 1 hour. The mixture was diluted with ethyl acetate, washed with saturated sodium chloride solution and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 100% EtOAc in hexane) to give 4-bromo-6-fluoro-7-methoxy-1H-indole. MS: (ES) m/z calculated for C ₉ H ₈ BrFNO [M+H] ⁺ 243.9, found 243.9.

Step b: To a suspension of 4-bromo-6-fluoro-7-methoxy-1H-indole (900 mg, 3.68 mmol), bis(pinacolato)diborone (1.21 g, 4.8 mmol) and KOAc (1.08 g, 11 mmol) in dioxane (16 ml) was added a complex of Pd(dppf)Cl ₂ with dichloromethane (400 mg, 0.49 mmol). The reaction mixture was degassed with nitrogen for 2 minutes and stirred at 100° C. for 2 hours. The reaction mixture was diluted with ethyl acetate and filtered through Celite. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 100% EtOAc in hexane) to give 6-fluoro-7-methoxy-4-(4.4 ,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole. MS: (ES) m/z calculated for C ₁₅ H ₂₀ BFNO ₃ [M+H] ⁺ 292.1, found 292.1.

Example 2

Synthesis of 4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c] pyrazol-3-yl)-6-fluoro-7-methoxy-1H-indole

Step a: To a 250 ml flask containing 90 ml concentrated hydrochloric acid was added 2,6-diethylaniline (10 g, 67 mmol) with magnetic stirring. The resulting mixture was stirred for 30 minutes and cooled in an ice/salt bath until the temperature inside the flask reached -5°C. A solution of sodium nitrite (5.5 g, 80 mmol) in water (60 ml) was slowly added to the mixture described above, keeping the temperature in the flask below 5°C.

Separately, tin(II) chloride dihydrate (31.6 g, 140 mmol) was added to a 500 ml 3-necked round bottom flask containing concentrated hydrochloric acid (60 ml) with mechanical stirring. The resulting solution was then cooled in an ice bath.

The diazonium salt slurry was then filtered into a 500 ml flask containing a chilled and vigorously stirred stannous chloride solution. After 90 minutes, the reaction mixture was transferred to a 500 ml Erlenmeyer flask and the flask was washed with water (20 ml) and chloroform (8 ml). The combined mixture was stirred overnight at room temperature. The liquid was decanted to give a wet solid. It was dried under vacuum for one day and then transferred to a 500 ml 3 neck round bottom flask equipped with a mechanical stirrer and stirred with diethyl ether (180 ml). The resulting mixture was cooled in an ice bath and slowly added 10N. NaOH solution (30 ml), keeping the temperature in the flask below 12°C. At the end of the addition, the mixture was left on ice for 2 hours. The ether layer was decanted into a 500 ml flask and a stream of hydrogen chloride gas was bubbled through the stirred ether solution. The precipitate that formed was filtered off to give (2,6-diethylphenyl)hydrazine hydrochloride. MS: (ES) m/z calculated for C ₁₀ H ₁₇ N ₂ [M+H] ⁺ 165.1, found 165.1.

Step b: Pyridine (4 ml, 49.5 mmol) was added to a mixture of (2,6-diethylphenyl)hydrazine hydrochloride (5 g, 24.91 mmol), tert-butyl 4-cyano-2,2-dimethyl-3-oxopyrrolidine-1- carboxylate (5 g, 20.98 mmol) and EtOH (60 ml) in a 250 ml round bottom flask with stirring on a magnetic stirrer. The resulting mixture was stirred at 70°C for 24 hours. The solvent was removed under reduced pressure, the residue was diluted with ethyl acetate and washed with aqueous citric acid solution, aqueous NaHCO ₃ solution, saturated sodium chloride solution and dried over MgSO ₄ . The solvent was removed under reduced pressure and the residue was crystallized from cyclohexane to give tert-butyl 3-amino-2-(2,6-diethylphenyl)-6,6-dimethyl-2,6-dihydropyrrolo[3,4-c]pyrazole- 5(4H)-carboxylate. MS: (ES) m/z calculated for C ₂₂ H ₃₃ N ₄ O ₂ [M+H] ⁺ 385.2, found 385.2.

Tert-butyl nitrite (0.5 ml, 3.8 mmol) was slowly added at room temperature to a mixture of tert-butyl 3-amino-2-(2,6-diethylphenyl)-6,6-dimethyl-2,6-dihydropyrrolo[3,4 -c]pyrazole-5(4H)-carboxylate (1 g, 2.6 mmol), diiodomethane (1.5 ml, 18.6 mmol) and MeCN (15 ml) in a 100 ml round bottom flask with stirring on a magnetic stirrer. The resulting mixture was stirred at 45°C for 3 hours, then diluted with toluene, washed with a saturated solution of NH ₄ Cl/NH ₄ OH (3:1), saturated sodium chloride solution and dried over MgSO ₄ . The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (2 to 25% EtOAc in hexanes) to give t-butyl 2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl- 2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate. MS: (ES) m/z calculated for C ₂₂ H ₃₁ IN ₃ O ₂ [M+H] ⁺ 496.1, found 496.2.

Step c: The above-described tert-butyl 2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate was dissolved in dichloromethane (10 ml) and HCl in dioxane (4N solution, 5 ml) was added. The resulting mixture was stirred at room temperature for 12 hours. Upon completion, the solvent was evaporated in vacuo to give 2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole hydrochloride. MS: (ES) m/z calculated for C ₁₇ H ₂₃ IN ₃ [M+H] ⁺ 396.1, found 396.2.

N,N-diisopropylethylamine (0.3 ml, 1.73 mmol) was added to a suspension of 2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c ]pyrazole hydrochloride (680 mg, 1.57 mmol), and 2,4-bis(trifluoromethyl)benzaldehyde (800 mg, 3.3 mmol) in 1,2-dichloroethane (10 mL) with magnetic stirring. After stirring at room temperature for 10 min, NaBH(OAc) ₃ (800 mg, 3.77 mmol) was added in portions. The resulting mixture was stirred at 45°C for 2 hours. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate, washed with aqueous NaHCO ₃ solution, saturated sodium chloride solution and dried over MgSO ₄ . The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (2 to 25% EtOAc in hexane) to give 5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl) -3-iodo-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole. MS: (ES) m/z calculated for C ₂₆ H ₂₇ F ₆ IN ₃ [M+H] ⁺ 622.1, found 622.1.

Step d: To a suspension of 5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3 ,4-c]pyrazole (250 mg, 0.40 mmol), 6-fluoro-7-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H- indole (100 mg, 0.34 mmol) and K ₂ CO ₃ (445 mg, 1.81 mmol) in dioxane (6 ml) and water (1 ml), a complex of Pd(dppf)Cl ₂ with dichloromethane (50 mg, 0.06 mmol) was added. The reaction mixture was degassed with nitrogen for 2 minutes and stirred under nitrogen at 100°C for 2.5 hours. The reaction mixture was diluted with ethyl acetate, washed with an aqueous solution of NaHCO ₃ and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (3 to 35% EtOAc in hexanes) then HPLC (MeCN/H ₂ O with 1% TFA) to give 4-(5-(2 ,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-6 -fluoro-7-methoxy-1H-indole. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.37 (broad s, 1H), 8.19 (d, J=12 Hz, 1H), 7.86 (s, 1H), 7.76 (d, J=12 Hz, 1H ), 7.07-7.33 (m, 4H), 6.41 (dd, J=2.2, 3.2 Hz, 1H), 6.33 (d, J=13.8 Hz, 1H), 4.14 (s, 2H), 4.01 (s, 3H) , 3.70 (s, 2H), 2.23-2.37 (m, 4H), 1.55 (s, 6H), 1.02 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₅ H ₃₄ F ₇ N ₄ O [M+H] ⁺ 659.2, found 659.2.

Example 3

Synthesis of 3-chloro-4-(2-(2,6-diethylphenyl)-6,6-dimethyl-5-(2-(trifluoromethyl)benzyl)-2,4,5,6-tetrahydropyrrolo[3,4-c ]pyrazol-3-yl)-7-fluoro-1H-indole

Step a: To a suspension of 4-bromo-7-fluoro-1H-indole (1.00 g, 4.67 mmol), bis(pinacolato)diborone (1.31 g, 5.14 mmol) and KOAc (1.15 g, 11.7 mmol) in dioxane (15 ml ) complex of Pd(dppf)Cl ₂ with dichloromethane (416 mg, 0.51 mmol) was added. The reaction mixture was degassed with nitrogen for 2 minutes and stirred at 100° C. for 2 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate and filtered through celite. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 30% EtOAc in hexane) to give 7-fluoro-4-(4, 4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole. MS: (ES) m/z calculated for C ₁₄ H ₁₈ BFNO ₂ [M+H] ⁺ 262.1, found 262.1.

Step b: To a suspension of tert-butyl 2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate (540 mg, 1.09 mmol), 7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (350 mg, 1.34 mmol) and K ₂ CO ₃ (830 mg, 6.78 mmol) in dioxane (10 ml) and water (2 ml) was added a complex of Pd(dppf)Cl ₂ with dichloromethane (200 mg, 0.32 mmol). The reaction mixture was degassed with nitrogen for 2 minutes and stirred under nitrogen at 100° C. for 2 hours. The mixture was cooled to room temperature, diluted with ethyl acetate, washed with aqueous NaHCO ₃ and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (5% to 25% EtOAc in hexane) to give t-butyl 2-(2,6-diethylphenyl)-3-(7-fluoro-1H-indole -4-yl)-6,6-dimethyl-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate. MS: (ES) m/z calculated for C ₃₀ H ₃₆ FN ₄ O ₂ [M+H] ⁺ 503.2, found 503.3.

Step c: The above-described tert-butyl 2-(2,6-diethylphenyl)-3-(7-fluoro-1H-indol-4-yl)-6,6-dimethyl-2,6-dihydropyrrolo[3,4- c]pyrazole-5(4H)-carboxylate (200 mg, 0.40 mmol) was dissolved in DMF (5 mL) and N-chlorosuccinimide (100 mg, 0.75 mmol) was added. The resulting mixture was stirred at room temperature for 12 hours. The reaction mixture was diluted with ethyl acetate, washed with aqueous Na ₂ S ₂ O ₃ solution, saturated sodium chloride solution and dried over MgSO ₄ . The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (2 to 25% EtOAc in hexane) to give t-butyl 3-(3-chloro-7-fluoro-1H-indol-4-yl)- 2-(2,6-diethylphenyl)-6,6-dimethyl-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate. MS: (ES) m/z calculated for C ₃₀ H ₃₃ ClFN ₄ O ₂ [M - H] ^- 535.2, found 535.2.

The above-described tert-butyl 3-(3-chloro-7-fluoro-1H-indol-4-yl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,6-dihydropyrrolo[3,4 -c]pyrazole-5(4H)-carboxylate was dissolved in dichloromethane (6 ml) and HCl in dioxane (4N solution, 5 ml) was added. The resulting mixture was stirred at room temperature for 12 hours. After completion of the reaction, the solvent was evaporated in vacuo to give 3-chloro-4-(2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole- 3-yl)-7-fluoro-1H-indole hydrochloride. MS: (ES) m/z calculated for C ₂₅ H ₂₇ ClFN ₄ [M+H] ⁺ 437.2, found 437.2.

Step d: N,N-diisopropylethylamine (0.2 ml, 1.15 mmol) was added to a suspension of 3-chloro-4-(2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo [3,4-c]pyrazol-3-yl)-7-fluoro-1H-indole hydrochloride (75 mg, 0.16 mmol) and 2-trifluoromethylbenzaldehyde (120 mg, 0.69 mmol) in 1,2-dichloroethane (6 ml) while stirring on a magnetic stirrer. After stirring at room temperature for 10 min, NaBH(OAc) ₃ (150 mg, 0.71 mmol) was added to the reaction mixture. The resulting mixture was stirred at 40°C for 1 hour, then cooled to room temperature, diluted with ethyl acetate, washed with saturated sodium chloride solution and dried over MgSO ₄ . The solvent was removed under reduced pressure and the residue was purified by preparative TLC (40% EtOAc in hexane) then HPLC (MeCN/H ₂ O with 1% TFA) to give 3-chloro-4-(2-(2.6 -diethylphenyl)-6,6-dimethyl-5-(2-(trifluoromethyl)benzyl)-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-7-fluoro-1H- indole ¹ H-NMR (400 MHz, CD ₃ OD) δ 7.92-7.99 (m, 1H), 7.52-7.66 (m, 2H), 7.34-7.38 (m, 2H), 7.24-7.29 (m, 2H), 6.94 -6.98 (m, 1H), 6.66 (dd, J=8.2, 10.8 Hz, 1H), 6.50 (dd, J=4.5, 8.2 Hz, 1H), 4.88 (broad, 1H), 4.09 (s, 2H) , 3.82 (d, J=11.5 Hz, 1H), 3.43 (d, J=11.5 Hz, 1H), 2.53 (d.sq., J=7.5, 15 Hz, 1H), 2.37-2.46 (m, 2H), 2.07 (d.sq., J=7.5, 15 Hz, 1H), 1.54 (s, 3H), 1.51 (s, 3H), 1.33 (t, J=7.6 Hz, 3H), 0.76 (t, J=7.6 Hz , 3H). MS: (ES) m/z calculated for C ₃₃ H ₃₂ ClF ₄ N ₄ [M+H] ⁺ 595.2, found 595.2.

Example 4

Synthesis of 4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c] pyrazol-3-yl)-7-fluoro-1H-indole

A mixture of 5-(2,4-bis(trifluoromethyl)benzyl)-3-iodo-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c ]pyrazole (200 mg, 0.32 mmol), 7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (150 mg, 0.57 mmol) , K ₂ CO ₃ (276 mg, 2.0 mmol) and Pd(dppf)Cl ₂ complex with dichloromethane (60 mg, 0.07 mmol) in dioxane (6 ml) and water (1 ml) were stirred at 100°C for 5 hours in a nitrogen atmosphere. The mixture was cooled to room temperature, diluted with ethyl acetate, and filtered through a pad of celite. The filtrate was collected, evaporated in vacuo and the residue was purified by silica gel flash chromatography (0 to 50% EtOAc in hexane) to give 4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2, 6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-7-fluoro-1H-indole. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.44 (s, 1H), 8.19 (d, J=8.0 Hz, 1H), 7.86 (s, 1H), 7.75 (d, J=8.4 Hz, 1H), 7.22 (m, 2H), 7.07 (d, J=7.6 Hz, 2H), 6.61 (m, 1H), 6.47 (m, 2H), 4.15 (s, 2H), 3.71 (s, 2H), 2.37 (m , 2H), 2.22 (m, 2H), 1.56 (s, 6H), 1.00 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₄ H ₃₂ F ₇ N ₄ [M+H] ⁺ 629.2, found 629.2.

Example 5

Synthesis of 4-(5-(3,5-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c] pyrazol-3-yl)-7-fluoro-1H-indole

Step a: Mixture of 2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole hydrochloride (2.20 g, 5.1 mmol), _{in _} DCM (30 ml) was stirred at room temperature for 1.5 hours. The reaction was then quenched with aqueous NaHCO ₃ and extracted with ethyl acetate. The organic layer was separated, dried over Na ₂ SO ₄ , evaporated in vacuo and the residue was purified by silica gel flash chromatography (0 to 30% EtOAc in hexane) to give 5-(3,5-bis(trifluoromethyl)benzyl)- 2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole. MS: (ES) m/z calculated for C ₂₆ H ₂₇ FIN ₃ [M+H] ⁺ 622.1, found 622.1.

Step b: A mixture of 5-(3,5-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3, 4-c]pyrazole (1.50 g, 2.4 mmol), 7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (817 mg, 3.1 mmol), K ₂ CO ₃ (0.830 g, 6.0 mmol) and the Pd(dppf)Cl ₂ complex with dichloromethane (30 mg, 0.36 mmol) in dioxane (12 mL) and water (1.5 mL) were stirred at 100°C in for 2 hours under nitrogen atmosphere. The reaction mixture was cooled to room temperature and partitioned between EtOAc and aqueous NaHCO ₃ solution. The organic layer was separated, dried over Na ₂ SO ₄ and evaporated in vacuo. The residue was purified by silica gel flash chromatography (0 to 50% EtOAc in hexanes) then HPLC (MeCN/H ₂ O with 1% TFA) to give 4-(5-(3,5-bis(trifluoromethyl) benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-7-fluoro-1H-indole. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.48 (s, 1H), 7.92 (s, 2H), 7.14 (s, 1H), 7.24 (dd, J=7.6, 7.6 Hz, 1H), 7.20 (d , J=2.8 Hz, 1H), 7.07 (dd, J=7.6, 7.6 Hz, 2H), 6.61 (m, 1H), 6.47 (m, 2H), 4.03 (s, 2H), 3.65 (s, 2H) , 2.37 (m, 2H), 2.22 (m, 2H), 1.56 (s, 6H), 1.00 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₄ H ₃₂ F ₇ N ₄ [M+H] ⁺ 629.2, found 629.2.

Example 6

Synthesis of methyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole-7-carboxylate intermediate

Step a: Into a suspension of methyl 4-bromo-1H-indole-7-carboxylate (300 mg, 1.18 mmol), bis(pinacolato)diborone (330 mg, 1.30 mmol) and KOAc (290 mg, 2.96 mmol) in dioxane (8 mL) a complex of Pd(dppf)Cl ₂ with dichloromethane (100 mg, 0.12 mmol) was added. The reaction mixture was degassed with nitrogen for 2 minutes and stirred at 100° C. for 1 hour. The reaction mixture was diluted with ethyl acetate, filtered through Celite. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (5 to 30% EtOAc in hexane) to give methyl 4-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)-1H-indole-7-carboxylate. MS: (ES) m/z calculated for C ₁₆ H ₂₁ BNO ₄ [M+H] ⁺ 302.2, found 302.2.

Example 7

Synthesis of (4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c ]pyrazol-3-yl)-1H-indol-7-yl)methanol

Step a: A mixture of 5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3, 4-c]pyrazole (100 mg, 0.16 mmol), methyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole-7-carboxylate (78 mg , 0.26 mmol), K ₂ CO ₃ (150 mg, 1.1 mmol) and Pd(dppf)Cl ₂ complex with dichloromethane (90 mg, 0.11 mmol) in dioxane (4 ml) and water (0.7 ml) were stirred at 100°C for 2 hours under nitrogen atmosphere. The reaction mixture was cooled to room temperature and partitioned between EtOAc and aqueous NaHCO ₃ solution. The organic layer was separated, dried over Na ₂ SO ₄ and evaporated in vacuo. The residue was purified by silica gel flash chromatography (0 to 40% EtOAc in hexane) to give methyl 4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6, 6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-1H-indole-7-carboxylate. MS: (ES) m/z calculated for C ₃₆ H ₃₅ F ₆ N ₄ O ₂ [M+H] ⁺ 669.3, found 669.3.

Step b: To a solution of methyl 4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3 ,4-c]pyrazol-3-yl)-1H-indole-7-carboxylate (25 mg, 0.037 mmol) in THF (1.5 ml) at 0°C was added a solution of LiAlH ₄ in diethyl ether (1.0 M, 0.20 ml, 0.20 mmol). After 1 hour at 0°C, the reaction was quenched with water and partitioned between EtOAc and aqueous NaHCO ₃ solution. The organic layer was separated, dried over Na ₂ SO ₄ and evaporated in vacuo. The residue was purified by HPLC (MeCN/H ₂ O, with 1% TFA), then silica gel flash chromatography (0 to 60% EtOAc in hexane) to give (4-(5-(2,4-bis(trifluoromethyl )benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-1H-indol-7-yl )methanol. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.99 (s, 1H), 8.18 (d, J=8.0 Hz, 1H), 7.86 (s, 1H), 7.74 (d, J=8.4 Hz, 1H), 7.24 (m, 2H), 7.06 (d, J=7.6 Hz, 2H), 6.70 (d, J=7.4 Hz, 1H), 6.49 (d, J=7.4 Hz, 1H), 6.48 (d, J=2.6 Hz, 1H), 4.92 (d, J=5.6 Hz, 2H), 4.15 (s, 2H), 3.72 (s, 2H), 2.39 (m, 2H), 2.26 (m, 2H), 1.83 (t, J =5.8 Hz, 1H), 1.56 (s, 6H), 1.02 (t, J=7.6, 6H). MS: (ES) m/z calculated for C ₃₅ H ₃₅ F ₆ N ₄ O [M+H] ⁺ 641.3, found 641.3.

Example 8

Synthesis of 3,3,3-trifluoro-2,2-dimethylpropanoyl chloride intermediate

A mixture of 3,3,3-trifluoro-2,2-dimethylpropanoic acid (0.312 g, 2.0 mmol), oxalyl chloride (0.17 ml, 2.0 mmol) and DMF (2 drops) in DCM (6.7 ml) was stirred at room temperature for 30 minutes. The reaction mixture containing 3,3,3-trifluoro-2,2-dimethylpropanoyl chloride was directly used in the next step without further purification.

Example 9

Synthesis of 4-(2-(2,6-diethylphenyl)-6,6-dimethyl-5-(3,3,3-trifluoro-2,2-dimethylpropyl)-2,4,5,6-tetrahydropyrrolo[3, 4-c]pyrazol-3-yl)-7-fluoro-1H-indole

Step a: A mixture of 4-(2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-7-fluoro- 1H-indole hydrochloride (78 mg, 0.18 mmol), 3,3,3-trifluoro-2,2-dimethylpropanoyl chloride (~2 mmol) and NEt₃(0.13 ml, 0.89 mmol) in DCM (3 ml) was stirred for 1 hour at room temperature. The mixture was then partitioned between EtOAc and aqueous NaHCO₃. The organic layer was separated, dried over Na₂SO_four and evaporated in vacuo. The residue was purified by silica gel flash chromatography (0 to 15% EtOAc in DCM) to give 1-(2-(2,6-diethylphenyl)-3-(7-fluoro-1H-indol-4-yl)-6 ,6-dimethyl-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)-3,3,3-trifluoro-2,2-dimethylpropan-1-one. MS: (ES) m/z calculated for C_thirtyH₃₃F_fourN_fourO[M+H]⁺541.3, found 541.2.

Step b: To a solution of 1-(2-(2,6-diethylphenyl)-3-(7-fluoro-1H-indol-4-yl)-6,6-dimethyl-2,6-dihydropyrrolo[3,4- c]pyrazol-5(4H)-yl)-3,3,3-trifluoro-2,2-dimethylpropan-1-one (35 mg, 0.064 mmol) in THF (2 ml) was added a solution of DIBAL-H in DCM ( 1.0 M, 1.5 ml, 1.5 mmol). After 1 hour at room temperature, the reaction was quenched with aqueous NaHCO ₃ and partitioned between EtOAc and aqueous NaHCO ₃ . The organic layer was separated, dried over Na ₂ SO ₄ and evaporated in vacuo. The residue was purified by silica gel flash chromatography (0 to 10% EtOAc in DCM) to give 4-(2-(2,6-diethylphenyl)-6,6-dimethyl-5-(3,3,3-trifluoro- 2,2-dimethylpropyl)-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-7-fluoro-1H-indole. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.49 (s, 1H), 7.27 (dd, J=2.4, 2.4 Hz, 1H), 7.23 (dd, J=8.0, 8.0 Hz, 1H), 7.06 (d , J=7.2 Hz, 2H), 6.63 (m, 1H), 6.56 (m, 1H), 6.47 (m, 1H), 3.93 (s, 2H), 2.84 (s, 2H), 2.35 (m, 2H) , 2.20 (m, 2H), 1.41 (s, 6H), 1.19 (s, 6H), 0.98 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₀ H ₃₅ F ₄ N ₄ [M+H] ⁺ 527.3, found 527.2.

Example 10

Synthesis of 4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c] pyrazol-3-yl)-3-fluoro-1H-indole

Step a: To a suspension of 4-bromo-3-fluoro-1H-indole (240 mg, 1.1 mmol), bis(pinacolato)diborone (420 mg, 1.7 mmol) and KOAc (320 mg, 3.3 mmol) in dioxane (5 ml ) complex of Pd(dppf)Cl ₂ with dichloromethane (140 mg, 0.17 mmol) was added. The reaction mixture was degassed with nitrogen for 2 minutes and stirred at 90° C. for 2 hours. The reaction mixture was adsorbed onto silica gel and purified by silica gel flash chromatography (0 to 100% EtOAc in hexane) to give 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl)-1H-indole. MS: (ES) m/z calculated for C ₁₄ H ₁₈ BFNO ₂ [M+H] ⁺ 262.1, found 261.2.

Step b: To a suspension of 5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3 ,4-c]pyrazole (74 mg, 0.12 mmol), 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (64 mg , 0.25 mmol) and K ₂ CO ₃ (169 mg, 1.2 mmol) in dioxane (6 ml) and water (1 ml), a complex of Pd(dppf)Cl ₂ with dichloromethane (49 mg, 0.060 mmol) was added. The reaction mixture was degassed with nitrogen for 2 minutes and stirred under nitrogen at 90° C. for 16 hours. The reaction mixture was adsorbed onto silica gel and purified by silica gel flash chromatography (0 to 30% MTBE in hexane), followed by HPLC (MeCN/H ₂ O, with 1% TFA) and preparative TLC (EtOAc in toluene) to give 4 -(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole- 3-yl)-3-fluoro-1H-indole. ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.25 (d, J=8.4 Hz, 1H), 7.89-7.94 (m, 2H), 7.28 (t, J=7.6 Hz, 1H), 7.20 (dd, J=8.0, 2.6 Hz, 1H), 7.09-7.15 (m, 3H), 6.85 (t, J=8.4 Hz, 1H), 6.51 (d, J=7.2 Hz, 1H), 4.19 (s, 2H), 3.65 (s, 2H), 2.19-2.50 (m, 4H), 1.54 (s, 6H), 0.87-1.21 (m, 6H). MS: (ES) m/z calculated for C ₃₄ H ₃₂ F ₇ N ₄ [M+H] ⁺ 629.3, found 629.3.

Example 11

Synthesis of 4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c] pyrazol-3-yl)-3-methyl-1H-indole

Step a: To a suspension of 4-bromo-3-methyl-1H-indole (240 mg, 1.1 mmol), bis(pinacolato)diborone (910 mg, 3.6 mmol) and KOAc (710 mg, 7.2 mmol) in dioxane (6 ml ) complex of Pd(dppf)Cl ₂ with dichloromethane (290 mg, 0.36 mmol) was added. The reaction mixture was degassed (N ₂ ) for 2 minutes and stirred at 90°C for 4 hours. The reaction mixture was adsorbed onto silica gel and purified by silica gel flash chromatography (0 to 25% MTBE in hexane) to give 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl)-1H-indole. MS: (ES) m/z calculated for C ₁₅ H ₂₁ BNO ₂ [M+H] ⁺ 258.2, found 258.1.

Step b: To a suspension of 5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-3-iodo-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3 ,4-c]pyrazole (65 mg, 0.10 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (127 mg , 0.49 mmol) and K ₂ CO ₃ (256 mg, 1.9 mmol) in dioxane (6 ml) and water (1 ml), a complex of Pd(dppf)Cl ₂ with dichloromethane (51 mg, 0.062 mmol) was added. The reaction mixture was degassed with nitrogen for 2 minutes and stirred under nitrogen at 90° C. for 2 hours. The reaction mixture was adsorbed onto silica gel and purified by flash chromatography on silica gel (0 to 100% MTBE in hexane), followed by HPLC (MeCN/H ₂ O, with 1% TFA) and preparative TLC (40% acetone in hexane), giving 4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,5,6-tetrahydropyrrolo[3,4-c] pyrazol-3-yl)-3-methyl-1H-indole. ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.25-8.18 (m, 1H), 7.92 (d, J=9.0 Hz, 2H), 7.29-7.17 (m, 3H), 7.04 (d, J=1.2 Hz, 1H), 6.97-6.90 (m, 1H), 6.77 (dd, J=8.2, 7.3 Hz, 1H), 6.44 (dd, J=7.4, 0.9 Hz, 1H), 4.25-4.10 (m, 2H) , 3.66 (d, J=11.6 Hz, 1H), 3.51 (d, J=11.6 Hz, 1H), 2.28-2.58 (m, 3H), 2.17 (s, 3H), 2.00-2.11 (m, 1H), 1.55 (s, 3H), 1.53 (s, 3H), 1.34 (t, J=7.6 Hz, 3H), 0.77 (t, J=7.6 Hz, 3H). MS: (ES) m/z calculated for C ₃₅ H ₃₅ F ₆ N ₄ [M+H] ⁺ 625.3, found 625.3.

Example 12

Synthesis of tert-butyl 2-(2,6-dimethylphenyl)-3-(1H-indol-5-yl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5-carboxylate

Step a: EtOH (160 ml) was added to tert-butyl 3-cyano-4-oxo-pyrrolidine-1-carboxylate (19.5 g, 92.54 mmol) and (2,6-dimethylphenyl)hydrazine hydrochloride (16 g, 92.65 mmol) and AcOH (40 ml). The resulting suspension was stirred at 50° C. overnight. After completion, the reaction was quenched with 1N. aqueous NaOH solution and was extracted with ethyl acetate (2 x 100 ml), dried (MgSO ₄ ) and evaporated in vacuum. The resulting crude product was then purified by silica gel flash chromatography (50% EtOAc in hexane) to give t-butyl 3-amino-2-(2,6-dimethylphenyl)-4,6-dihydropyrrolo[3,4-c]pyrazole -5-carboxylate. MS: (ES) m/z calculated for C ₁₈ H ₂₅ N ₄ O ₂ [M+H] ⁺ 329.2, found 329.2.

Step b: Isoamyl nitrite (11.74 ml, 87.5 mmol) was added slowly at room temperature to a mixture of tert-butyl 3-amino-2-(2,6-dimethylphenyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5 -carboxylate (14.35 g, 43.75 mmol), diiodomethane (14 ml, 175 mmol) and MeCN (180 ml). The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was adsorbed onto silica gel and purified by silica gel flash chromatography (30% EtOAc in hexane) to give tert-butyl 2-(2,6-dimethylphenyl)-3-iodo-4,6-dihydropyrrolo[3,4-c ]pyrazole-5-carboxylate. MS: (ES) m/z calculated for C ₁₈ H ₂₃ IN ₃ O ₂ [M+H] ⁺ 440.1, found 440.2.

Step c: To a suspension of tert-butyl 2-(2,6-dimethylphenyl)-3-iodo-4,6-dihydropyrrolo[3,4-c]pyrazole-5-carboxylate (878 mg, 2 mmol), 5-( 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (729 mg, 3 mmol) and Na ₂ CO ₃ (530 g, 5 mmol) in dioxane (8 ml ) and water (2 ml) were added a complex of Pd(dppf)Cl ₂ with dichloromethane (163 mg, 0.2 mmol). The reaction mixture was degassed with nitrogen for 2 minutes and stirred at 90° C. for 2 hours. The reaction mixture was diluted with ethyl acetate and filtered through celite. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (0 to 100% EtOAc in hexanes) to give tert-butyl 2-(2,6-dimethylphenyl)-3 -(1H-indol-5-yl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5-carboxylate. MS: (ES) m/z calculated for C ₂₆ H ₂₉ N ₄ O ₂ [M+ H] ⁺ 429.2, found 429.2.

Example 13

The compounds in Table 1 and Table 2 below were prepared according to the methods described above. Characteristic data (MS and/or NMR) are given for each compound in the list.

Table 1 Structure and NMR/MS Data for private embodiments Structure ¹ H-NMR MS

Etc. 13.001

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.44 (d, J=3.1 Hz, 1H), 7.32 (t, J=8.0 Hz, 1H), 7.16 (d, J=7.6 Hz, 2H), 6.59 -6.65 (m, 1H), 6.53-6.56 (m, 1H), 6.45-6.50 (m, 1H), 4.72 (s, 2H), 3.88 (s, 2H), 2.19-2.37 (m, 4H), 1.87 (s, 6H), 1.46 (s, 9H), 0.99 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₂ H ₃₉ FN ₅ O ₃ [M+H] ⁺ 560.3, found 560.3. Etc. 13.002

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.89 (broad s, 1H), 7.37 (d, J=3.2 Hz, 1H), 7.31 (t, J=8.0 Hz, 1H), 7.12-7.15 ( m, 3H), 6.54-6.60 (m, 1H), 6.50-6.53 (m, 1H), 6.39-6.43 (m, 1H), 4.12 (s, 2H), 3.90 (s, 2H), 2.15-2.37 ( m, 4H), 1.50 (s, 6H), 0.97 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₂₉ H ₃₁ FN ₅ O [M+H] ⁺ 484.3, found 484.3. Etc. 13.003

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.53 (s, 1H), 7.27 (dd, J=2.6, 2.6 Hz, 1H), 7.23 (dd, J=7.6, 7.6 Hz, 1H), 7.06 (d , J=7.6, 2H), 6.63 (m, 1H), 6.56 (m, 1H), 6.48 (m, 1H), 3.99 (s, 2H), 2.70 (s, 2H), 2.39 (m, 2H), 2.23 (m, 2H), 1.62 (broad s, 1H), 1.43 (s, 6H), 1.25 (s, 6H), 0.99 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₂₉ H ₃₆ FN ₄ O [M+H] ⁺ 475.3, found 475.3. Etc. 13.004

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.20-8.27 (m, 1H), 7.89-7.92 (m, 2H), 7.38 (s, 1H), 7.22-7.32 (m, 2H), 6.95-6.98 (m, 1H), 6.66 (dd, J=8.2, 10.8 Hz, 1H), 6.50 (dd, J=4.5, 8.2 Hz, 1H), 4.88 (broad, 1H), 4.18 (s, 2H), 3.84 (d, J=11.4 Hz, 1H), 3.49 (d, J=11.4 Hz, 1H), 2.47 (d.sq., J=7.5, 15 Hz, 1H), 2.37-2.46 (m, 2H), 2.07 ( d.sq., J=7.5, 15 Hz, 1H), 1.54 (s, 3H), 1.52 (s, 3H), 1.33 (t, J=7.6 Hz, 3H), 0.76 (t, J=7.6 Hz, 3H ). MS: (ES) m/z calculated for C ₃₄ H ₃₁ ClF ₇ N ₄ [M+H] ⁺ 663.2, found 663.2. Etc. 13.005

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.56-7.61 (m, 1H), 7.34 (d, J=3.0 Hz, 1H), 7.30 (t, J=7.6 Hz, 1H), 7.18-7.22 ( m, 1H), 7.14 (d, J=8.0 Hz, 2H), 7.01-7.07 (m, 1H), 6.53-6.58 (m, 1H), 6.45 (t, J=3.1 Hz, 1H), 6.38-6.42 (m, 1H), 4.04 (s, 2H), 3.73 (s, 2H), 2.15-2.38 (m, 4H), 1.57 (s, 6H), 0.98 (t, J=7.2 Hz, 6H). MS: (ES) m/z calculated for C ₃₂ H ₃₂ ClF ₂ N ₄ [M+H] ⁺ 545.2, found 545.2. Etc. 13.006

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.50 (m, 1H), 7.37 - 7.26 (m, 2H), 7.14 (d, J=7.7 Hz, 2H), 6.91 (t, J=8.7 Hz, 2H), 6.55 (dd, J=10.9, 8.2 Hz, 1H), 6.48 - 6.36 (m, 2H), 3.96 (s, 2H), 3.73 (s, 2H), 2.26 (m, 4H), 1.56 (s , 6H), 0.97 (t, J=7.6, 6H). MS: (ES) m/z calculated for C ₃₂ H ₃₂ F ₃ N ₄ [M+H] ⁺ 529.3, found 529.3. Etc. 13.007

¹ H-NMR salt with TFA (400 MHz, CD ₃ OD) δ 11.41 (s, 1H), 8.75 (d, J=5.6 Hz, 2H), 7.94 (d, J=6.4 Hz, 2H), 7.39 (dd , J=2.6, 2.6 Hz, 1H), 7.34 (dd, J=7.6, 7.6 Hz, 1H), 7.16 (d, J=7.6 Hz, 2H), 6.60 (m, 1H), 6.45 (m, 2H) , 4.67 (s, 2H), 4.39 (s, 2H), 2.25 (m, 4H), 1.88 (s, 6H), 0.98 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₁ H ₃₃ FN ₅ [M+H] ⁺ 494.3, found 494.3. Etc. 13.008

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.47-7.57 (m, 1H), 7.39 (s, 1H), 7.23-7.28 (m, 2H), 7.12-7.16 (m, 2H), 6.95-6.98 (m, 1H), 6.66 (dd, J=8.2, 10.8 Hz, 1H), 6.50 (dd, J=4.5, 8.2 Hz, 1H), 4.88 (broad, 1H), 4.00 (d, J=11.4 Hz , 1H), 3.81-3.90 (m, 2H), 3.50 (d, J=11.4 Hz, 1H), 2.33-2.46 (m, 3H), 2.00-2.13 (m, 1H), 1.56 (s, 3H), 1.49 (s, 3H), 1.31 (t, J=7.6 Hz, 3H), 0.75 (t, J=7.6 Hz, 3H). MS: (ES) m/z calculated for C ₃₂ H ₃₁ Cl ₂ F ₂ N ₄ [M+H] ⁺ 579.2, found 579.2. Etc. 13.009

¹ H-NMR salt with TFA (400 MHz, CD ₃ OD) δ 11.46 (s, 1H), 8.87 (d, J=1.0 Hz, 1H), 8.71 (d, J=5.2 Hz, 1H), 8.26 (d , J=7.6 Hz, 1H), 7.68 (m, 1H), 7.41 (dd, J=2.8, 2.8 Hz, 1H), 7.35 (dd, J=7.8, 7.8 Hz, 1H), 7.18 (d, J= 8.0 Hz, 2H), 6.61 (m, 1H), 6.45 (m, 2H), 4.75 (s, 2H), 4.53 (s, 2H), 2.25 (m, 4H), 1.97 (s, 6H), 0.97 ( t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₁ H ₃₃ FN ₅ [M+H] ⁺ 494.3, found 494.3. Etc. 13.010

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.46-7.52 (m, 1H), 7.39 (s, 1H), 7.23-7.28 (m, 2H), 6.82-7.00 (m, 3H), 6.66 (dd , J=8.2, 10.8 Hz, 1H), 6.50 (dd, J=4.5, 8.2 Hz, 1H), 4.88 (broad, 1H), 4.00 (d, J=11.6 Hz, 1H), 3.81-3.90 (m , 2H), 3.50 (d, J=11.6 Hz, 1H), 2.30-2.58 (m, 3H), 2.00-2.12 (m, 1H), 1.57 (s, 3H), 1.49 (s, 3H), 1.31 ( t, J=7.6 Hz, 3H), 0.75 (t, J=7.6 Hz, 3H). MS: (ES) m/z calculated for C ₃₂ H ₃₁ ClF ₃ N ₄ [M+H] ⁺ 563.2, found 563.2. Etc. 13.011

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.26-7.34 (m, 3H), 7.16 - 7.25 (m, 2H), 7.13 (d, J=7.2 Hz, 2H), 6.90-7.00 (m, 1H ), 6.55 (dd, J=10.9, 8.1 Hz, 1H), 6.36-6.46 (m, 2H), 5.46-5.51 (m, 1H), 3.93 (s, 2H), 3.67 (s, 2H), 2.05- 2.45 (m, 4H), 1.54 (s, 6H), 0.98 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₂ H ₃₃ F ₂ N ₄ [M+H] ⁺ 511.3, found 511.3. Etc. 13.012

¹ H-NMR salt with TFA (400 MHz, CD ₃ OD) δ 11.46 (s, 1H), 8.68 (d, J=4.8 Hz, 1H), 7.92 (dd, J=1.6, 7.6 Hz, 1H), 7.62 (d, J=8.0, 1H), 7.45 (m, 2H), 7.38 (dd, J=7.6, 7.6 Hz, 1H), 7.18 (d, J=7.6 Hz, 2H), 6.62 (m, 1H), 6.53 (m, 1H), 6.45 (m, 1H), 4.79 (s, 2H), 4.64 (s, 2H), 2.27 (m, 4H), 1.92 (s, 6H), 0.98 (t, J=7.6 Hz , 6H). MS: (ES) m/z calculated for C ₃₁ H ₃₃ FN ₅ [M+H] ⁺ 494.3, found 494.3. Etc. 13.013

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.57 (d, J=8.2 Hz, 1H), 7.44 (d, J=2.1 Hz, 1H), 7.26-7.37 (m, 3H), 7.14 (d, J=7.7 Hz, 2H), 6.66 (dd, J=8.2, 10.8 Hz, 1H), 6.39-6.46 (m, 2H), 4.88 (broad, 1H), 4.04 (s, 2H), 3.74 (s, 2H), 2.03-2.33 (m, 4H), 1.56 (s, 6H), 0.98 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₂ H ₃₂ Cl ₂ FN ₄ [M+H] ⁺ 561.2, found 561.2. Etc. 13.014

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.42 (d, J=8.4 Hz, 2H), 7.28-7.34 (m, 4H), 7.20-7.26 (m, 1H), 7.14 (d, J=7.7 Hz, 2H), 6.55 (dd, J=8.2, 11.0 Hz, 1H), 6.47 - 6.35 (m, 2H), 3.93 (s, 2H), 3.68 (s, 2H), 2.12-2.39 (m, 4H) , 1.56 (s, 6H), 0.97 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₂ H ₃₄ FN ₄ [M+H] ⁺ 493.3, found 493.3. Etc. 13.015

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.32-7.49 (m, 1H), 7.25-7.34 (m, 3H), 7.04-7.14 (m, 4H), 6.53-6.57 (m, 1H), 6.38 -6.46 (m, 2H), 4.88 (broad, 1H), 3.99 (s, 2H), 3.74 (s, 2H), 2.17-2.36 (m, 4H), 1.57 (s, 6H), 0.97 (t, J=7.6Hz, 6H). MS: (ES) m/z calculated for C ₃₂ H ₃₃ F ₂ N ₄ [M+H] ⁺ 511.3, found 511.3. Etc. 13.016

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.39-7.45 (m, 2H), 7.26-7.36 (m, 2H), 7.14 (d, J=8.2 Hz, 2H), 6.98-7.04 (m, 2H ), 6.55 (dd, J=8.2, 11.0 Hz, 1H), 6.35-6.46 (m, 2H), 4.88 (broad, 1H), 3.91 (s, 2H), 3.66 (s, 2H), 2.17-2.36 (m, 4H), 1.55 (s, 6H), 0.97 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₂ H ₃₃ F ₂ N ₄ [M+H] ⁺ 511.3, found 511.3. Etc. 13.017

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.23 (d, J=8.2 Hz, 1H), 7.92-7.93 (m, 2H), 7.27-7.39 (m, 2H), 7.15 (d, J=7.7 Hz, 2H), 6.83 (d, J=7.7 Hz, 1H), 6.42-6.48 (m, 2H), 4.88 (broad, 1H), 4.22 (s, 2H), 3.66 (s, 2H), 2.20- 2.36 (m, 4H), 1.56 (s, 6H), 1.00 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₄ H ₃₂ ClF ₆ N ₄ [M+H] ⁺ 645.2, found 645.2. Etc. 13.018

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.24 (d, J=8.6 Hz, 1H), 7.89 - 7.95 (m, 2H), 7.24 - 7.34 (m, 2H), 7.13 (d, J=7.7 Hz, 2H), 6.60 (d, J=7.2 Hz, 1H), 6.33 - 6.42 (m, 2H), 4.22 (s, 2H), 3.73 (s, 2H), 2.16 - 2.42 (m, 7H), 1.56 (s, 6H), 0.99 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₅ H ₃₅ F ₆ N ₄ [M+H] ⁺ 625.3, found 625.3. Etc. 13.019

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.25-8.29 (m, 1H), 7.95-7.97 (m, 2H), 7.21-7.29 (m, 4H), 7.07 (d, J=1.4 Hz, 1H ), 6.47 (dd, J=1.4, 12.6 Hz, 1H), 6.33-6.35 (m, 1H), 4.88 (broad, 1H), 4.25 (s, 2H), 3.93 (s, 2H), 2.26-2.35 (m, 4H), 1.53 (s, 6H), 1.04 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₄ H ₃₂ F ₇ N ₄ [M+H] ⁺ 629.2, found 629.3. Etc. 13.020

¹ H-NMR salt with TFA (400 MHz, CD ₃ OD) δ 11.31 (s, 1H), 8.56 (d, J=4.4 Hz, 1H), 7.99 (dd, J=1.6, 7.2 Hz, 1H), 7.73 (d, J=7.6 Hz, 1H), 7.50 (m, 1H), 7.32 (m, 2H), 7.15 (d, J=7.6 Hz, 2H), 6.58 (m, 1H), 6.44 (m, 1H) , 6.39 (m, 1H), 4.71 (s, 2H), 2.28 (m, 4H), 2.03 (s, 6H), 0.99 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₁ H ₃₁ FN ₅ O[M+H] ⁺ 508.2, found 508.2. Etc. 13.021

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.30 (s, 1H), 7.91 (d, J=7.6 Hz, 1H), 7.51 (d, J=8.0 Hz, 1H), 7.40 (dd, J=7.6 , 7.6 Hz, 1H), 7.14 (m, 3H), 6.97 (d, J=7.6 Hz, 2H), 6.51 (m, 1H), 6.41 (m, 1H), 6.36 (m, 1H), 4.00 (s , 2H), 3.62 (s, 2H), 2.28 (m, 2H), 2.12 (m, 2H), 1.47 (s, 6H), 0.90 (t, J=7.4 Hz, 6H). MS: (ES) m/z calculated for C ₃₃ H ₃₃ F ₄ N ₄ [M+H] ⁺ 561.3, found 561.3. Etc. 13.022

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.54 (s, 1H), 7.26 (dd, J=2.8, 2.8 Hz, 1H), 7.16 (dd, J=7.6, 7.6 Hz, 1H), 6.99 (d , J=7.6 Hz, 2H), 6.59 (m, 1H), 6.43 (m, 2H), 4.53 (s, 2H), 2.20 (m, 6H), 1.84 (s, 6H), 0.91 (m, 13H) . MS: (ES) m/z calculated for C ₃₀ H ₃₆ FN ₄ O [M+H] ⁺ 487.3, found 487.3. Etc. 13.023

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.19 (s, 1H), 8.11 (d, J=8.0 Hz, 1H), 7.78 (s, 1H), 7.67 (d, J=8.4 Hz, 1H), 7.17 (dd, J=7.8, 7.8Hz, 1H), 7.09 (dd, J=2.8, 2.8Hz, 1H), 7.01 (d, J=7.6Hz, 2H), 6.81 (dd, J=2.0, 8.8Hz , 1H), 6.35 (m, 1H), 6.25 (dd, J=2.0, 10.8 Hz, 1H), 4.06 (s, 2H), 3.64 (s, 2H), 2.28 (m, 2H), 2.15 (m, 2H), 1.48 (s, 6H), 0.92 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₄ H ₃₂ F ₇ N ₄ [M+H] ⁺ 629.2, found 629.2. Etc. 13.024

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.54 (s, 1H), 7.34 (dd, J=2.8, 2.8 Hz, 1H), 7.26 (m, 1H), 7.07 (d, J=7.6 Hz, 2H ), 6.68 (m, 1H), 6.49 (m, 2H), 4.85 (s, 2H), 2.84 (s, 2H), 2.32 (m, 2H), 2.20 (m, 2H), 1.91 (s, 6H) , 1.54 (s, 6H), 1.00 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₀ H ₃₃ F ₄ N ₄ O [M+H] ⁺ 541.3, found 541.2. Etc. 13.025

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.21 (d, J=8.4 Hz, 1H), 7.90 (broad s, 2H), 7.24 - 7.31 (m, 4H), 6.92 - 6.95 (m, 1H ), 6.88 (t, J=8.4 Hz), 6.55 (dd, J=0.76, 7.6 Hz, 1H), 4.18 (s, 2H), 3.86 (d, J=12 Hz, 1H), 3.48 (d, J =11.6 Hz, 1H), 2.03 - 2.61 (m, 4H), 1.54 (s, 3H), 1.53 (s, 3H), 1.35 (t, J=7.6 Hz, 3H), 0.75 (t, J=7.6 Hz , 3H). MS: (ES) m/z calculated for C ₃₄ H ₃₂ ClF ₆ N ₄ [M+H] ⁺ 645.2, found 645.2. Etc. 13.026

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.38 (d, J=8.7 Hz, 1H), 8.06 (s, 1H), 7.88 (d, J=4.5 Hz, 2H), 7.41 (dd, J= 8.3, 0.8 Hz, 1H), 7.23 - 7.33 (m, 2H), 6.93-7.08 (m, 2H), 6.68-6.75 (m, 1H), 4.21 (s, 2H), 3.99 (d, J=11.3 Hz) , 1H), 3.51 (d, J=11.4 Hz, 1H), 2.46-2.68 (m, 2H), 2.30 - 2.40 (m, 1H), 1.98 - 2.06 (m, 1H), 1.57 (s, 3H), 1.55 (s, 3H), 1.36 (t, J=7.5 Hz, 3H), 0.77 (t, J=7.6 Hz, 3H). MS: (ES) m/z calculated for C ₃₅ H ₃₂ F ₆ N ₅ [M+H] ⁺ 636.3, found 636.3. Etc. 13.027

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.42 (s, 1H), 8.20 (d, J=8.4 Hz, 1H), 7.86 (s, 1H), 7.77 (d, J=8.0 Hz, 1H), 7.21 (m, 2H), 7.03 (broad s, 2H), 6.51 (t, J=10.4 Hz, 1H), 6.38 (m, 1H), 4.14 (s, 2H), 3.63 (s, 2H), 2.38 (broad s, 4H), 1.56 (s, 6H), 1.00 (broad s, 6H). MS: (ES) m/z calculated for C ₃₄ H ₃₁ F ₈ N ₄ [M+H] ⁺ 647.2, found 647.2. Etc. 13.028

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.34 (s, 1H), 7.89 (d, J=8.2 Hz, 1H), 7.69-7.77(m, 1H), 7.22-7.35 (m, 3H), 7.14 (d, J=7.7 Hz, 2H), 6.80 (t, J=7.8 Hz, 1H), 6.49 (m, 1H), 6.34 (m, 1H), 4.22 (s, 2H), 3.71 (s, 2H ), 2.20 - 2.50 (m, 4H), 1.56 (s, 6H), 0.99 (m, 6H). MS: (ES) m/z calculated for C ₃₄ H ₃₃ F ₆ N ₄ [M+H] ⁺ 611.3, found 611.3. Etc. 13.029

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.38 (d, J=3.0 Hz, 1H), 7.24-7.35 (m, 4H), 7.15 (d, J=7.6 Hz, 2H), 6.90-7.01 ( m, 3H), 6.85 (t, J=7.6 Hz, 1H), 6.54 (d, J=7.2 Hz, 1H), 6.48 (d, J=3.1 Hz, 1H), 4.82 (s, 2H), 2.18- 2.42 (m, 4H), 1.91 (s, 6H), 0.99 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₃ H ₃₅ N ₄ O ₂ [M+H] ⁺ 519.3, found 519.3. Etc. 13.030

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.63 (d, J=2.7 Hz, 1H), 7.20-7.40 (m, 5H), 7.13 (d, J=7.7 Hz, 2H), 6.76-6.85 ( m, 1H), 6.49 (dt, J=7.5, 0.7 Hz, 1H), 6.40 (dd, J=3.2, 0.8 Hz, 1H), 4.06 (s, 2H), 3.76 (s, 2H), 3.31 (s , 2H), 2.15-2.45 (m, 4H), 1.57 (s, 6H), 0.94 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₂ H ₃₃ Cl ₂ N ₄ [M+H] ⁺ 543.2, found 543.2. Etc. 13.031

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.23 (d, J=8.2 Hz, 1H), 7.92-7.93 (m, 2H), 7.22-7.39 (m, 3H), 7.07 (broad, 2H) , 6.65 (dd, J=8.2, 10.8, 1H), 6.23-6.27 (m, 1H), 4.87 (broad, 1H), 4.21 (s, 2H), 3.67 (s, 2H), 2.20-2.36 (b ., 4H), 1.57 (s, 6H), 0.88–1.02 (broad, 6H). MS: (ES) m/z calculated for C ₃₄ H ₃₂ F ₇ N ₄ [M+H] ⁺ 629.2, found 629.3. Etc. 13.032

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.24 (d, J=7.9 Hz, 1H), 7.54-7.64 (m, 2H), 7.23-7.42 (m, 4H), 7.13 (d, J=7.7 Hz, 2H), 6.80 (t, J=7.7 Hz, 1H), 6.35-6.49 (m, 2H), 4.86 (broad, 1H), 4.13 (s, 2H), 3.70 (s, 2H), 2.17- 2.42 (m, 4H), 1.55 (s, 6H), 0.99 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₃ H ₃₄ F ₃ N ₄ [M+H] ⁺ 543.3, found 543.6. Etc. 13.033

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.24 (d, J=8.2 Hz, 1H), 7.90-7.93 (m, 2H), 7.26-7.32 (m, 3H), 7.14 (d, J=7.7 Hz, 2H), 6.80 (t, J=7.7 Hz, 1H), 6.35-6.49 (m, 2H), 4.88 (broad, 1H), 4.22 (s, 2H), 3.74 (s, 2H), 2.20- 2.36 (m, 4H), 1.56 (s, 6H), 0.99 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₄ H ₃₃ F ₆ N ₄ [M+H] ⁺ 611.3, found 611.6. Etc. 13.034

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.68 (s, 1H), 7.37-7.46 (m, 2H), 7.14-7.32 (m, 5H), 7.07 (d, J=1.4 Hz, 1H), 6.84-6.86 (m, Hz, 1H), 6.25-6.26 (m, 1H), 4.86 (broad, 1H), 4.08 (s, 2H), 3.94 (s, 2H), 2.26-2.35 (m, 4H) , 1.53 (s, 6H), 1.03 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₂ H ₃₃ Cl ₂ N ₄ [M+H] ⁺ 543.2, found 543.5. Etc. 13.035

¹ H-NMR (400 MHz, CD ₃ OD) δ 8.01 (d, J=8.0 Hz, 1H), 7.58-7.73 (m, 2H), 7.42 (t, J=7.7 Hz, 2H), 7.13-7.28 ( m, 5H), 6.82 (dd, J=1.7, 8.0 Hz, 1H), 6.23-6.24 (m, 1H), 4.88 (broad, 1H), 4.16 (s, 2H), 3.88 (s, 2H), 2.20-2.36 (m, 4H), 1.52 (s, 6H), 1.03 (t, J=7.6 Hz, 6H). MS: (ES) m/z calculated for C ₃₃ H ₃₄ F ₃ N ₄ [M+H] ⁺ 543.3, found 543.5. Etc. 13.036

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.13 (broad, 1H), 7.32-7.37 (m, 1H), 7.15-7.22 (m, 4H), 7.06 (d, J=7.6 Hz, 1H), 6.91 (dd, J=1.7, 8.5 Hz, 1H), 6.43-6.44 (m, 1H), 4.08 (s, 2H), 2.50 (s, 2H), 2.00 (s, 6H), 1.36 (s, 6H) , 1.00 (s, 9H). MS: (ES) m/z calculated for C ₂₈ H ₃₅ N ₄ [M+H] ⁺ 427.3, found 427.5. Etc. 13.037

¹ H-NMR (400 MHz, CD ₃ OD) δ 7.55-7.63 (m, 2H), 7.10-7.39 (m, 9H), 6.82-6.90 (m, 1H), 6.23-6.24 (m, 1H), 4.82 (broad, 2H), 4.64 (d, J=11.5 Hz, 1H), 4.33 (d, J=11.5 Hz, 1H), 3.87 (s, 1H), 2.06-2.39 (m, 4H), 1.59 (s , 3H), 1.35 (s, 3H), 1.19 (s, 3H), 1.05 (s, 3H), 0.83–1.03 (m, 6H). MS: (ES) m/z calculated for C ₃₅ H ₄₁ N ₄ O [M+H] ⁺ 533.3, found 533.7. Etc. 13.038

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.15 (broad, 1H), 7.48-7.56 (m, 2H), 7.24-7.44 (m, 5H), 7.08-7.20 (m, 4H), 6.83 (dd , J=1.7, 8.6 Hz, 1H), 6.36-6.37 (m, 1H), 4.93 (s, 1H), 4.69 (d, J=11.4 Hz, 1H), 4.18-4.35 (m, 4H), 3.94 ( e, J=11.4 Hz, 1H), 2.14-2.43 (m, 4H), 1.58 (s, 3H), 1.51 (s, 3H), 1.19-1.34 (m, 6H), 0.96-1.07 (m, 6H) . MS: (ES) m/z calculated for C ₃₅ H ₃₉ N ₄ O ₂ [M+H] ⁺ 547.3, found 547.5. Etc. 13.039

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.17 (broad, 1H), 7.13-7.37 (m, 4H), 7.03-7.06 (m, 2H), 6.89 (dd, J=1.1, 8.5 Hz, 1H ), 6.42–6.44 (m, 1H), 4.18 (s, 2H), 4.06 (s, 2H), 2.69 (s, 2H), 2.00 (s, 6H), 1.01 (s, 9H). MS: (ES) m/z calculated for C ₂₆ H ₃₁ N ₄ [M+H] ⁺ 399.3, found 399.5.

table 2 Structure and MC Data for Private Embodiments Structure MS

Etc. 13.040

MS: (ES) m/z calculated for C ₂₉ H ₃₅ FN ₅ O [M+H] ⁺ 488.3, found 488.3. Etc. 13.041

MS: (ES) m/z calculated for C ₃₅ H ₃₂ F ₆ N ₄ O ₂ [M+H] ⁺ 655.2, found 655.2. Etc. 13.042

MS: (ES) m/z calculated for C ₂₇ H ₃₁ FN ₅ O [M+H] ⁺ 460.3, found 460.3. Etc. 13.043

MS: (ES) m/z calculated for C ₃₄ H ₃₃ F ₆ N ₄ O [M+H] ⁺ 627.3, found 627.3. Etc. 13.044

MS: (ES) m/z calculated for C ₃₂ H ₃₄ F ₃ N ₆ O [M+H] ⁺ 575.3, found 575.3. Etc. 13.045

MS: (ES) m/z calculated for C ₃₄ H ₃₃ F ₆ N ₄ O [M+H] ⁺ 627.3, found 627.3. Etc. 13.046

MS: (ES) m/z calculated for C ₃₀ H ₃₇ N ₄ O ₂ [M+H] ⁺ 485.3, found 485.5. Etc. 13.047

MS: (ES) m/z calculated for C ₂₅ H ₃₀ N ₅ [M+H] ⁺ 400.2, found 400.5. Etc. 13.048

MS: (ES) m/z calculated for C ₂₇ H ₃₁ N ₄ O ₂ [M+H] ⁺ 443.2, found 443.2. Etc. 13.049

MS: (ES) m/z calculated for C ₂₆ H ₂₇ N ₄ O ₃ [M+H] ⁺ 443.2, found 443.2. Etc. 13.050

MS: (ES) m/z calculated for C ₂₉ H ₃₃ N ₄ O [M+H] ⁺ 453.2, found 453.2. Etc. 13.051

MS: (ES) m/z calculated for C ₂₈ H ₃₁ N ₄ O [M+H] ⁺ 439.2, found 439.2. Etc. 13.052

MS: (ES) m/z calculated for C ₂₅ H ₂₈ N ₅ O ₂ [M+H] ⁺ 430.2, found 430.2. Etc. 13.053

MS: (ES) m/z calculated for C ₂₇ H ₂₉ N ₄ O [M+H] ⁺ 425.2, found 425.2. Etc. 13.054

MS: (ES) m/z calculated for C ₃₀ H ₃₅ N ₄ O [M+H] ⁺ 467.3, found 467.5. Etc. 13.055

MS: (ES) m/z calculated for C ₂₇ H ₃₁ N ₄ O [M+H] ⁺ 427.2, found 427.2. Etc. 13.056

MS: (ES) m/z calculated for C ₂₆ H ₂₉ N ₄ O ₂ [M+H] ⁺ 429.2, found 429.5. Etc. 13.057

MS: (ES) m/z calculated for C ₃₁ H ₃₃ N ₄ O ₂ S [M+H] ⁺ 525.2, found 525.2. Etc. 13.058

MS: (ES) m/z calculated for C ₃₂ H ₃₃ N ₄ O [M+H] ⁺ 489.3, found 489.3. Etc. 13.059

MS: (ES) m/z calculated for C ₂₆ H ₃₀ N ₅ O [M+H] ⁺ 428.2, found 428.2. Etc. 13.060

MS: (ES) m/z calculated for C ₂₈ H ₂₅ N ₄ O [M+H] ⁺ 433.2, found 433.2.

Example 14

This example illustrates the evaluation of the biological activity of particular variants of the compounds of the present invention.

Materials and methods

A. Cells

1. Cells expressing C5a receptors

a) U937 cells

U937 cells are a line of monocytic leukocytes that express C5aR and are available from the American Type Culture Collection (ATCC (VA)). These cells were grown in 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 (fetal bovine serum). Cells were grown in 5% CO ₂ /95% air, 100% humidity at 37°C, subcultured twice a week at a ratio of 1:6 (cells were grown at a density ranging from 1 x 10 ⁵ to 2 x 10 ⁶ cells /ml) and harvested at a concentration of 1 × 10 ⁶ cells/ml. Prior to analysis, 0.5 mM cyclic AMP (Sigma, OH) was added to cells overnight and washed once before use. cAMP-treated U937 cells were used in C5aR ligand binding assays and in functional assays.

b) Isolated human neutrophils

Optionally, human or rodent neutrophils can be used to test the activity of the compounds. Neutrophils can be isolated from fresh human blood by density separation and centrifugation techniques. Briefly, whole blood is incubated with equal parts of 3% dextran and allowed to separate for 45 minutes. After separation of the layers, the top layer is placed on top of 15 ml Ficoll (15 ml Ficoll for every 30 ml of blood suspension) and centrifuged for 30 minutes at 400 x g without stopping. The pellet at the bottom of the tube is removed and resuspended in PharmLyse RBC lysis buffer (BD Biosciences, San Jose, CA), after which the sample is centrifuged again for 10 minutes at 400 x g without stopping. The resulting pellet of cells is re-suspended as necessary, it consists of isolated neutrophils.

B. Tests

1. Suppression of ligand binding to C5aR

CAMP-treated U937 cells expressing C5aR were centrifuged and resuspended in assay buffer (20 mM HEPES pH 7.1, 140 mM NaCl, 1 mM CaCl ₂ , 5 mM MgCl ₂ , with 0.1% bovine serum albumin) at a concentration of 3 × 10 ⁶ cells/ml. The binding assay was performed as follows. 0.1 ml of cell suspension was added to assay plates containing 5 µl of compounds, resulting in a final concentration of ~2-10 µM of each screened compound (or part of the dose-response relationship in determining the IC ₅₀ value for this compound). Then 0.1 ml of ¹²⁵ I-labeled C5a (obtained from Perkin Elmer Life Sciences, Boston, MA) diluted in assay buffer to a final concentration of ~50 pM, giving ~30,000 cpm/well, was added, the plates were sealed and incubated about 3 hours at 4°C on a shaker platform. The reaction solutions were aspirated on GF/B glass filters soaked in 0.3% polyethyleneimine (PEI) solution into a vacuum cell collector (Packard Instruments; Meriden, CT). Scintillation fluid (40 μl; Microscint 20, Packard Instruments) was added to each well, the plates were sealed, and the radioactivity level was measured in a Topcount scintillation counter (Packard Instruments). Control wells containing either diluent alone (to assess overall pulse levels) or excess C5a (1 μg/mL, for non-specific binding) were used to calculate percent total inhibition for each compound. The computer program Prism from GraphPad, Inc. was used. (San Diego, Ca) to calculate IC ₅₀ values. The IC ₅₀ values are the concentrations required to reduce the binding of radiolabeled C5a to the receptor by 50%. (For more information on 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)).

1. Mobilization of calcium

Optionally, compounds can be tested for their ability to inhibit calcium flux in cells. To detect calcium release from intracellular stores, cells (eg, cAMP-stimulated U937 or neutrophils) are incubated with 3 μM INDO-1AM dye (Molecular Probes; Eugene, OR) in cell growth medium for 45 minutes at room temperature and washed with phosphate - saline buffer (PBS). After INDO-1AM loading, cells were resuspended in flow assay buffer (Hanks Balanced Salt Solution (HBSS) and 1% FBS). Calcium mobilization was measured using a Photon Technology International spectrophotometer (Photon Technology International; New Jersey) with an excitation wavelength of 350 nm and double simultaneous detection of emitted fluorescence at a wavelength of 400 nm and 490 nm. The relative concentrations of intracellular calcium are expressed as the emission ratio of 400 nm/490 nm. Experiments are carried out at 37° C. with continuous agitation in cuvettes each containing 10 ⁶ cells in 2 ml of flow analysis buffer. Chemokine ligands can be used in the concentration range from 1 to 100 nM. The emission ratio is plotted on a chart against time (usually 2-3 minutes). Candidate compounds blocking ligand binding (up to 10 μM) are added at time 10 seconds, followed by addition of chemokines at time 60 seconds (C5a; R&D Systems; Minneapolis, MN) and control chemokine (SDF-1α; R&D Systems; Minneapolis , MN) at time 150 seconds.

2. Chemotaxis analysis

Optionally, compounds can be tested for their ability to inhibit chemotaxis in cells. Chemotaxis assays are performed with polyvinylpyrrolidone-coated polycarbonate filters with a pore size of 5 μm in 96-well chemotaxis assay chambers (Neuroprobe; Gaithersburg, MD) using a chemotaxis assay buffer (Hanks Balanced Salt Solution (HBSS) and 1% FBS) . C5aR ligands (C5a, R&D Systems; Minneapolis, MN) were used to evaluate compound inhibition of C5aR mediated migration. Other chemokines (SDF-1α; R&D Systems; Minneapolis, MN) are used to control specificity. 29 μl of chemokine (0.03 nM C5a) and varying amounts of the compound are placed in the lower chamber; the upper chamber contains 100,000 U937 cells or 20 µl of neutrophils. The chambers are incubated for 1.5 hours at 37°C and the number of cells in the lower chamber is quantified either by direct cell count in five fields of view at high magnification, or by CyQuant (Molecular Probes) - a fluorescent staining method that determines the content of nucleic acid and gives a microscopic picture.

C. Identification of C5aR inhibitors

1. Analysis

To test for small organic molecules that interfere with C5a receptor binding to ligand, an assay that detects a radioactive ligand (eg, C5a) bound to cells expressing C5aR on the surface (eg, cAMP-stimulated U937 cells or isolated human neutrophils) was used. For compounds that inhibit binding, competitively or non-competitively, lower radioactivity values are observed compared to uninhibited controls.

Equal numbers of cells were added to each well in the plate. The cells were then incubated with radiolabeled C5a. Unbound ligand was removed by washing the cells, and bound ligand was determined by quantitative counting of the number of pulses. Cells that were incubated without adding any organic compound gave the total number of pulses (ppm total); non-specific binding was determined by incubating cells with unlabeled ligand and labeled ligand. The percentage of inhibition was determined by the equation:

% inhibition=(1 - [(imp/min in sample) - (non-specific imp/min)]/[(imp/min total) - (non-specific imp/min)]) × 100.

2. Dose-response curves

To determine the affinity of candidate compounds for C5aR, as well as confirm their ability to inhibit ligand binding, their inhibitory activity was determined in the concentration range from 1 × 10 ^-10 to 1 × 10 ^-4 M. In this study, the amount of the compound was varied; the number of white blood cells and the concentration of the ligand remained constant.

D. In vivo efficacy model

Test compounds were evaluated for their potential efficacy in the treatment of C5a-mediated conditions by determining the efficacy of the compound in an animal model. In addition to the models described below, other suitable animal models for testing these compounds can be found in Mizuno, M. et al., Expert Opin. Investig. Drugs (2005), 14(7), 807-821, which is hereby incorporated by reference in its entirety.

1. Models of C5a-induced leukopenia

a) C5a-induced leukopenia in a knock-in mouse model with the human C5aR gene

To test the efficacy of the compounds of the present invention in an animal model, recombinant mice were generated by standard procedures in which the genetic sequence encoding mouse C5aR was changed to that encoding human C5aR, resulting in hC5aR-KI mice. In these mice, administration of hC5a leads to increased production of adhesion molecules on the walls of blood vessels that bind white blood cells, removing them from the bloodstream. Animals were injected with 20 μg/kg hC5a, and after 1 minute the number of leukocytes in the peripheral blood was determined according to standard methods. Preliminary administration to mice of various dosages of the compounds of the present invention can almost completely block hC5a-induced leukopenia.

b) C5a-induced leukopenia in a cynomolgus monkey (Cynomolgus) model

To investigate the efficacy of the compounds of the present invention in a non-human simian model, C5a-induced leukopenia was investigated in a cynomolgus (cynomolgus) model. In this model, administration of hC5a leads to increased production of adhesion molecules on the walls of blood vessels that bind white blood cells, removing them from the bloodstream. Animals were injected with 10 μg/kg hC5a and after 1 minute the number of leukocytes in the peripheral blood was determined.

Mouse model of ANCA-induced vasculitis

On Day 0, hC5aR-KI mice were intravenously injected with 50 mg/kg of purified anti-myeloperoxidase antibodies (Xiao et al, J. Clin. Invest. 110: 955-963 (2002)). The mice were then given oral doses of the compounds of the present invention or placebo once a day for seven days, then the mice were sacrificed and the kidneys were taken for histological examination. Analysis of sections of the kidneys shows a significant reduction in the number and severity of crescentic and necrotic lesions in the glomeruli, compared with animals that were injected with placebo.

2. Mouse model of choroidal neovascularization

To investigate the efficacy of the compounds of the present invention in the treatment of age-related macular degeneration (AMD), Bruch's membrane in the eyes of hC5aR-KI mice was ruptured by laser photocoagulation (Nozika et al, PNAS 103: 2328-2333 (2006). Mice were administered orally or a placebo or a compound of the present invention was injected into the vitreous at appropriate dosages for 1 to 2 weeks Healing of laser-induced lesions and neovascularization was determined by histology and angiography.

3. Models of rheumatoid arthritis

a) Rabbit model of destructive joint inflammation

To investigate the effectiveness of the compounds of the present invention in inhibiting the inflammatory response in rabbits to intra-articular injection of LPS liposaccharide (a component of a bacterial membrane), a rabbit model of destructive joint inflammation was used. This study models the destructive joint inflammation seen in arthritis. Intra-articular injection of LPS elicits an acute inflammatory response characterized by the release of cytokines and chemokines, many of which are seen in rheumatoid arthritis joints. There is a sharp increase in the concentration of leukocytes in the synovial fluid and in the synovial membrane in response to the appearance of these chemotactic mediators. Selective chemokine receptor antagonists have shown efficacy in this model (see Podolin, et al., J. Immunol. 169(11):6435-6444 (2002)).

LPS study in rabbits was carried out according to the methods described in the above work by Podolin et al. Female New Zealand rabbits (weighing about 2 kilograms) were injected intra-articularly in one knee with LPS (10 ng) or together with placebo (phosphate-buffered saline with 1% DMSO) , or with the addition of a candidate compound (dosage 1=50 μM, or dosage 2=100 μM), in a total volume of 1.0 ml. 16 hours after LPS injection, the knees were washed and cell counts were performed. The positive effect of the administration of the compounds was confirmed by histopathological examination of synovial inflammation. Histopathological examination used a scoring system for assessing inflammation: 1 - minimal, 2 - weak, 3 - moderate, 4 - average.

b) Testing compounds in a rat model of collagen-induced arthritis

A 17-day collagen-induced type II arthritis study was conducted to evaluate the effect of a candidate compound on arthritis-induced clinical ankle swelling. Collagen-induced arthritis in the rat is an experimental model of polyarthritis that is widely used in preclinical testing of numerous antirheumatic drugs (see Trentham et al., J. Exp. Med. 146(3):857-868 (1977), Bendele et al. , Toxicologic Pathol 27:134-142 (1999), Bendele et al, Arthritis Rheum 42:498-506 (1999)). The hallmarks of this model are reliable onset and progression of sustained, easily measurable, multi-articular inflammation, severe cartilage destruction coupled with pannus formation, mild to moderate bone resorption, and pericartilaginous bone proliferation.

Female Lewis rats (weighing approximately 0.2 kg) were euthanized with isoflurane and injected with incomplete Freund's adjuvant containing 2 mg/ml bovine type II collagen at the base of the tail and at two sites on the back on days 0 and 6 of this 17 day study. The candidate compound was administered daily by subcutaneous injection from day 0 to day 17 at an effective dosage. The ankle diameter was measured with a caliper, and the reduction in ankle swelling was taken as a measure of effectiveness.

4. Rat model of sepsis

To investigate the efficacy of the compounds of the present invention in inhibiting the generalized inflammatory response associated with sepsis-like diseases, a cecal ligation and puncture (CLP) sepsis rat model was used. The study of rats with CLP was performed as described in Fujimura N, et al. (American Journal of Respiratory Critical Care Medicine 2000; 161: 440-446). Briefly, Wistar albino rats of both sexes weighing 200-250 grams were fasted for 12 hours prior to the experiment. Animals were kept under normal conditions with alternating 12 hour periods of light and dark and were fed standard rat chow for up to 12 hours prior to the start of the experiment. Then the animals were divided into four groups; (i) two sham groups and (ii) two CLP groups. Each of these two groups (ie, (i) and (ii)) was divided into a control group (placebo administered) and a test compound group. Sepsis was called by the CLP method. Under short anesthesia, a midline laparotomy was performed with minimal incision, and the caecum was ligated just below the ileocecal valve with 3-0 silk suture so as to maintain intestinal continuity. The antimesenteric surface of the caecum was perforated with a 18-gauge needle in two places at a distance of 1 cm from each other, and the caecum was gently squeezed until the fecal mass was squeezed out. The intestine was then returned to the abdominal cavity and the incision was sutured. At the end of the operation, all rats were resuscitated with saline, 3 ml/100 grams of body weight, subcutaneously. After the operation, the rats were not given food, but they had unlimited access to water for 16 hours, after which the rats were slaughtered. The sham groups underwent laparotomy and caecal manipulation, but no ligation or perforation. The positive effect of the administration of the test compounds was determined by histopathological scoring of tissues and organs, as well as measurements of several key indicators of liver, kidney and lipid peroxidation. Aspartate transaminase (AST) and alanine transaminase (ALT) levels were measured to assess liver function. To assess the work of the kidneys, the concentrations of urea nitrogen and creatinine in the blood were studied. The blood levels of pro-inflammatory cytokines, such as TNF-alpha and IL-1beta, were also analyzed by ELISA.

5. Mouse SLE model of experimental lupus nephritis.

An MRL/lpr mouse SLE model was used to study the effect of the compounds of the present invention on systemic lupus erythematosus (SLE). The MRL/Mp-Tmfrsf6 ^lpr/lpr (MRL/lpr) line is a widely used model of human SLE. To test the efficacy of the compounds in this model, male MRL/lpr mice are evenly divided between the control group and the C5aR antagonist group at 13 weeks of age. Compound or placebo is then administered via osmotic pumps for the next 6 weeks to minimize stress effects in animals. Blood and urine samples are taken every 2 weeks during the 6 weeks of disease onset and progression. A minority of these mice develop glomerulosclerosis, leading to death of the animal due to kidney failure. Mortality as an indicator of kidney failure is one of the criteria measured, and successful treatment usually results in a reduction in the number of sudden deaths in test groups. In addition, the presence and severity of kidney damage can also be continuously monitored by measurements of blood urea nitrogen and urine protein. Tissues and organs were also harvested at 19 weeks of age, histopathological and immunohistochemical studies were performed, and tissue damage and cell infiltration were assessed on a scoring scale.

6. Rat model of COPD

Smoke-induced inflammation of the airways in a rat model can be used to evaluate the efficacy of compounds in chronic obstructive pulmonary disease (COPD). Selective chemokine antagonists have shown efficacy in this model (see Stevenson, et al., Am. J. Physiol Lung Cell Mol Physiol. 288 L514-L522, (2005)). A rat model of acute COPD was created as described by Stevenson et al. The test compound was administered either systemically orally or intravenously, or applied topically via a nebulizer. Male Sprague-Dawley rats (350-400 g) were placed in Perspex chambers and exposed to cigarette smoke via a pump (50 ml every 30 seconds, with fresh air pauses). The total exposure time in rats was 32 minutes. Rats were sacrificed 7 days after initial exposure. The positive effect of the administration of the compounds was assessed by a decrease in the infiltration of inflammatory cells, a decrease in the levels of chemokines and cytokines.

In the chronic model, mice or rats were exposed to daily tobacco smoke for up to 12 months. The compound was administered systemically once a day orally or topically via a nebulizer. In addition to the inflammation observed in the acute model (according to Stevensen et al.), other pathologies similar to those observed in humans with COPD, such as emphysema (indicated by midlinear intercept) and changes in lung chemistry, can also be observed in animals (see Martorana). et al, Am J Respir Crit Care Med 172(7): 848-53.

7. Mouse EAE model of multiple sclerosis

Experimental autoimmune encephalomyelitis (EAE) is a model of human multiple sclerosis. Variations of this model have been published and are well known in the art. In a typical procedure, C57BL/6 mice (Charles River Laboratories) are used to create an EAE model. Mice are immunized with 200 µg of myelin oligodendrocyte glycoprotein (MOG) 35-55 (Peptide International) emulsified in Freund's complete adjuvant (CFA) containing 4 mg/ml Mycobacterium tuberculosis (Sigma-Aldrich) subcutaneously on day 0. In addition, at day 0 and day 2 animals are injected with 200 ng of pertussis toxin (Calbiochem) intravenously. Clinical symptoms are assessed on a scale from 0 to 5 points: 0, no signs of disease; 1, flaccid tail; 2, hind limb weakness; 3, hind limb paralysis; 4, weakness or paralysis of the forelimbs; 5, a dying animal. The administration of the test compound begins on day 0 (prophylactic) or day 7 (therapeutic, when there is histological evidence of the presence of the disease, but clinical symptoms are observed in a small number of animals), and is administered once or more times a day at concentrations suitable for assessing their activity and pharmacokinetic parameters, eg 100 mg/kg subcutaneously. The efficacy of the compounds can be assessed by comparing the severity (maximum mean score in the presence of the compound versus placebo) or by measuring the reduction in the number of macrophages (F4/80 positive) shed from the spinal cord. Spinal cord mononuclear cells can be isolated with a discontinuous Percoll gradient. Cells can be stained with rat anti-mouse F4/80-PE or rat IgG2b-PE (Caltag Laboratories) and quantified by FACS analysis using 10 µl of Polybeads per sample (Polysciences).

8. Mouse model of kidney transplantation

Transplantation models can be created in mice, for example, the allogeneic kidney transplant model from C57BL/6 to BALB/c mice is described in Faikah Gueler et al, JASN Express, Aug 27th, 2008. Briefly, mice are anesthetized and the left kidney attached to the to the cuff of the aorta and the renal vein with a small hollow cuff, with the ureters. After removal of the left kidney from the recipient, the cuffs of the vessels are connected by anastomosis to the recipient's abdominal aorta and vena cava, respectively, below the level of native renal vessels. The ureter is directly connected by an anastomosis to the bladder. The critical graft cold storage time is 60 minutes and the critical graft warm storage time is 30 minutes. The right native kidney can be removed at the time of allograft transplantation or on day 4 post-transplantation for long-term survival studies. Monitor the general physical condition of the mice as an indicator of rejection. The administration of the test compound to the animals can be started before surgery or immediately after transplantation, for example by subcutaneous injection once a day. Monitor kidney and animal function and survival. Plasma creatinine levels are monitored by an automated method (Beckman Analyzer, Krefeld, Germany).

9. Mouse model of ischemia/reperfusion

A mouse model of ischemia-reperfusion injury can be created as described by Xiufen Zheng et al, Am. J. Pathol, Vol 173:4, Oct, 2008. Briefly, 6-8 week old CD1 mice are anesthetized and placed on a warm mat to maintain temperature during surgery. After the abdominal wall incision, the renal pedicles are excised and a microvascular clip is placed on the left renal pedicle for 25-30 minutes. After ischemia, the clips are removed along with the right kidney, the incision is sutured, and the animal is left to recover. They take blood for the analysis of the concentration of creatinine and urea nitrogen as indicators of kidney health. Alternatively, animal survival is monitored over time. The compound can be administered to animals before and/or after surgery; and the effect on creatinine and urea nitrogen concentrations, as well as on animal survival, is used as an indicator of compound efficacy.

10. Mouse model of tumor growth

C57BL/6 mice aged 6-16 weeks are injected subcutaneously with 1×10 ⁵ TC-1 cells (ATCC, VA) in the left or right flank. 2 weeks after the injection of cells begin to measure the size of the tumor with a caliper every 2-4 days until the tumor reaches a size that requires the slaughter of the animal. At slaughter, the animals are opened and the spleen and tumors are taken. The excised tumors are measured and weighed. Compounds can be administered before and/or after tumor cell injection; and inhibition or inhibition of tumor growth is used to evaluate the effectiveness of the compound.

Table 3 below lists the structures and activities of representative compounds described in this text. Inhibition activity in the chemotaxis assay (see Example 14 B.3) is characterized as follows: +, 500 nM ≤ IC50;++, 50 nM ≤ IC50<500 nM; +++, 5 nM ≤ IC50<50 nM; and++++, IC50<5 nM.

Despite the description in the text of particular embodiments of the present invention, when reading the description, those skilled in the art may recognize variations of the disclosed embodiments, and those skilled in the art may implement such variations as necessary. Accordingly, the present invention may be practiced in ways other than those specifically disclosed in this application, and the present invention includes all modifications and equivalents to the subject matter described in the appended claims, in accordance with applicable law. In addition, any combination of the elements described above in all their possible variations are included in the scope of the present invention, unless otherwise specified or clearly follows from the context.

All publications, patent applications, account numbers, and other references cited in this text are incorporated herein by reference, as if each individual publication or patent application were separately and specifically indicated as incorporated by reference in the text.

Claims (67)

1. Compound having formula (I)

or a pharmaceutically acceptable salt thereof, wherein cycle member A ¹ is selected from the group consisting of N and CH; cycle member A ² is selected from the group consisting of N, CH and C(R ⁴ ); each of the cycle members A ³ , A ⁴ , A ⁵ and A ⁶ is independently selected from the group consisting of CH and C(R ⁴ ); each of the dotted lines independently represents a single or double bond; R ¹ is selected from the group consisting of -C _1-8 alkylene-heteroaryl, -C ₁ alkylene-C ₆ aryl, C _1-8 alkyl, -C(O)-C _1-8 alkyl, -C(O)- C ₆ aryl, -C(O)-heteroaryl, -C(O)-C _3-6 cycloalkyl, -C(O)NR ^1a R ^1b , SO ₂ -C ₆ aryl, -C(O)-C ₁ alkylene -OC ₆ aryl, and -C(O)-C _1-3 alkylene-C _3-6 cycloalkyl and -CO ₂ R ^1a ; a heteroaryl group is a 5-6 membered aromatic ring containing as ring members 1-2 heteroatoms selected from N and O; where R ^1a and R ^1b are each independently selected from the group consisting of a hydrogen atom and C _1-8 alkyl; where R ¹ is optionally substituted with 1-2 substituents R ⁵ ; R ^2a and R ^2e are each C _1-6 alkyl; R ^2b , R ^2c and R ^2d each represents a hydrogen atom; each R ³ is C _1-6 alkyl; each R ⁴ is independently selected from the group consisting of C _1-6 alkyl, C _1-6 alkoxy, C _1-6 hydroxyalkyl, halogen, cyano, and -CO ₂ R ^4a ; each R ^4a represents a hydrogen atom; each R ⁵ is independently selected from the group consisting of C _1-8 alkyl, C _1-8 haloalkyl, C _1-8 hydroxyalkyl, C _3-6 cycloalkyl, halogen, OH, -NR ^5a R ^5b , -NR ^5a C(O ) ₂ R ^5b and CO ₂ R ^5a ; where each R ^5a and R ^5b is independently selected from the group consisting of a hydrogen atom and C _1-8 alkyl; and subscript n is 0, 1, or 2. 2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ and A ⁶ as ring members is a bicyclic heteroaryl selected from the group consisting of from

where m is 0, 1, 2 or 3; and where the R ⁴ substituents may be attached to any suitable carbon atom in the bicyclic heteroaryl. 3. The compound according to any one of claims 1 or 2, or a pharmaceutically acceptable salt thereof, wherein the ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ and A ⁶ as ring members is a bicyclic heteroaryl represented by structure

, where m is 0, 1, 2 or 3; and where the R ⁴ substituents may be attached to any suitable carbon atom in the bicyclic heteroaryl. 4. A compound according to any one of claims 1 or 3, or a pharmaceutically acceptable salt thereof, wherein each R ⁴ is independently selected from the group consisting of C _1-4 alkyl, C _1-4 alkoxy, C _1-6 hydroxyalkyl, halogen, cyano groups and -CO ₂ R ^4a ; and where the R ⁴ substituents may be attached to any suitable carbon atom in the bicyclic heteroaryl. 5. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein the ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ and A ⁶ as ring members is a bicyclic heteroaryl selected from the group consisting of:

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein the ring fragment containing A ¹ , A ² , A ³ , A ⁴ , A ⁵ and A ⁶ as ring members is a bicyclic heteroaryl selected from the group consisting of:

7. A compound according to any one of claims 1-6 or a pharmaceutically acceptable salt thereof, wherein R ¹ is selected from the group consisting of -C _1-8 alkylene heteroaryl, -C ₁ alkylene-C ₆ aryl, C _1-8 alkyl, -C(O)-C _1-8 alkyl, -C(O)-C ₆ aryl, -C(O)-heteroaryl, -C(O)-C _3-6 cycloalkyl, -C(O)NR ^1a R ^1b , -SO ₂ -C ₆ aryl and -C(O)-C ₁ alkylene-OC ₆ aryl; where the heteroaryl group is a 5-6 membered aromatic ring containing as ring members 1-2 heteroatoms selected from N and O; and where R ^1a and R ^1b are each independently selected from the group consisting of a hydrogen atom and C _1-8 alkyl; and R ¹ is optionally substituted with 1-2 R ⁵ substituents. 8. A compound according to any one of claims 1-7, or a pharmaceutically acceptable salt thereof, wherein each R ⁵ is independently selected from the group consisting of C _1-8 alkyl, C _1-8 haloalkyl, C _1-8 hydroxyalkyl, C _3-6 cycloalkyl, halogen, OH, -NR ^5a R ^5b and CO ₂ R ^5a where each R ^5a and R ^5b is independently selected from the group consisting of hydrogen and C _1-8 alkyl. 9. A compound according to any one of claims 1-8, or a pharmaceutically acceptable salt thereof, wherein R ¹ is -CH ₂ -phenyl optionally substituted with 1-3 R ⁵ substituents. 10. A compound according to any one of claims 1-9, or a pharmaceutically acceptable salt thereof, wherein R ¹ is -CH ₂ -phenyl substituted with 1 or 2 R ⁵ substituents, where each R ⁵ is independently C _1-4 haloalkyl. 11. A compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein R ¹ is -CH ₂ -phenyl substituted with 1 or 2 CF ₃ substituents. 12. The compound according to any one of claims 1-7, or a pharmaceutically acceptable salt thereof, where R ¹ is selected from the group consisting of

13. A compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein R ¹ is

. 14. A compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein R ^2a and R ^2e are each independently selected from the group consisting of methyl and ethyl. 15. A compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein

selected from the group consisting of

. 16. A compound according to any one of claims 1-15, or a pharmaceutically acceptable salt thereof, wherein n is 2. 17. A compound according to any one of claims 1-16, or a pharmaceutically acceptable salt thereof, wherein each R ³ is independently C _1-4 alkyl. 18. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, wherein

selected from the group consisting of

. 19. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, having the structure represented by formula (Ia), (Ib) or (Ic):

20. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, having the structure represented by formula (Id), (Ie) or (If):

where m is 0, 1 or 2; where the substituents R ⁴ can be attached to any suitable carbon atom in bicyclic heteroaryl; and where p is 0.1 or 2. 21. A compound according to claim 2, or a pharmaceutically acceptable salt thereof, having a structure represented by formula (Ig) or (Ih):

where m is 0, 1 or 2, and where R ⁴ substituents may be attached to any suitable carbon atom of the indole heteroaryl. 22. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is

. 23. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is

. 24. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is

. 25. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is

. 26. The compound according to any one of claims 1 to 25, or a pharmaceutically acceptable salt thereof, selected from the group consisting of

27. Pharmaceutical composition for the treatment of diseases and disorders occurring with pathological activation of the C5a receptor, containing an effective amount of the compound according to any one of claims 1 to 26 or a pharmaceutically acceptable salt thereof as a C5a receptor modulator and a pharmaceutically acceptable carrier. 28. A method of treating a mammal suffering from or susceptible to a disease or disorder with pathological C5a activation, wherein the disease or disorder is selected from the group consisting of the following: C3 glomerulopathy, C3 glomerulonephritis, dense sediment disease, mesangioproliferative glomerulonephritis, hemolytic uremic syndrome, atypical hemolytic uremic syndrome (aHUS), ANCA-associated vasculitis, Wegener's granulomatosis, microscopic polyangiitis, autoimmune hemolytic and thrombocytopenic pathological conditions, immunovasculitis and glomerulonephritis, comprising administering to a mammal an effective amount of a compound according to any one of claims 1 to 26, or a pharmaceutically acceptable salt thereof, or compositions according to claim 27.