HK1166321B - 3-cycloalkylaminopyrrolidine derivatives as modulators of chemokine receptors - Google Patents

Description

3-cycloalkylaminopyrrolidine derivatives as modulators of chemokine receptors

The present application is a divisional application of patent application No. 200480037737.5 filed on 2004, 12/16/2004 entitled "3-cycloalkylaminopyrrolidine derivatives as chemokine receptor modulators".

Technical Field

The present invention relates to chemokine receptor modulators, e.g. antagonists, and their use as pharmaceutical ingredients. The invention further relates to novel compounds and medical methods of treating inflammation and other disorders, particularly those associated with lymphocyte or monocyte accumulation, such as rheumatoid arthritis, lupus, graft-versus-host disease, and/or transplant rejection. More specifically, the present invention relates to 3-cycloalkylaminopyrrolidine derivatives and their use as modulators of chemokine receptors.

More particularly, the present invention relates to novel compounds and pharmaceutical compositions thereof having anti-inflammatory and immunomodulatory biological activity that act via antagonism of the CCR2 receptor (also known as the MCP-1 receptor), thus leading to inhibition of monocyte chemoattractant protein-1 (MCP-1). The novel compounds are 3-cycloalkylaminopyrrolidine derivatives. The invention further relates to novel compounds for use in the compositions, processes for their preparation, intermediates useful in their preparation and their use as therapeutic agents.

The chemokine receptor modulators/antagonists of the present invention can be effective as therapeutic and/or prophylactic agents for the following diseases: such as atherosclerosis, asthma, pulmonary fibrosis, myocarditis, ulcerative colitis, psoriasis, asthma, ulcerative colitis, nephritis (nephropathy), multiple sclerosis, lupus, systemic lupus erythematosus, hepatitis, pancreatitis, sarcoidosis, organ transplantation, crohn's disease, endometriosis, congestive heart failure, viral meningitis, cerebral infarction, neuropathy, kawasaki disease, and sepsis, in which tissue infiltration of blood leukocytes, such as monocytes and lymphocytes, plays a major role in the initiation, progression, or maintenance of the disease.

The invention also provides compounds and pharmaceutical compositions thereof having immunomodulatory biological activity that act via antagonism of the CCR5 receptor.

Background

The migration and transport of leukocytes from blood vessels to diseased tissues appears to be a key element in the initiation of a normal disease-fighting inflammatory response. This process, also known as leukocyte recruitment, is also involved in the initiation and progression of life-threatening inflammatory diseases as well as debilitating autoimmune diseases. The pathogenesis of these diseases derives from the attack of the body's immune system defenses against normal tissues. Thus, preventing and blocking leukocyte recruitment to target tissues in inflammatory and autoimmune diseases would be a very effective therapeutic intervention.

Different leukocyte classes involved in cellular immune responses include monocytes, lymphocytes, neutrophils, eosinophils and basophils. In most cases, lymphocytes are the leukocyte class that initiates, coordinates, and maintains chronic inflammatory responses, and thus are generally the most important class of cells that block access to inflammatory sites. Lymphocytes attract monocytes to the tissue site, and together with lymphocytes, they are responsible for most of the actual tissue damage that occurs in inflammatory diseases. Infiltration of lymphocytes and/or monocytes is known to cause a wide range of chronic autoimmune diseases and organ transplant rejection. These diseases include, but are not limited to, rheumatoid arthritis, chronic contact dermatitis, inflammatory bowel disease, lupus, systemic lupus erythematosus, multiple sclerosis, atherosclerosis, psoriasis, sarcoidosis, idiopathic pulmonary fibrosis, dermatomyositis, cutaneous pemphigoid and related diseases (e.g., pemphigus vulgaris, pemphigus foliaceus, pemphigus rubra), glomerulonephritis, vasculitis, hepatitis, diabetes, allograft rejection, and graft-versus-host disease.

The process of leukocyte depletion from the bloodstream and accumulation at inflammatory sites and initiation of disease has at least three steps, which are described as (1) fluctuations, (2) activated/firm adhesion, and (3) transendothelial migration [ Springer, t.a., Nature 346: 425, 433 (1990); lawrence and Springer, Cell 65: 859-873 (1991); butcher, e.c., Cell 67: 1033-1036(1991)]. The second step is mediated at the molecular level by chemoattractant receptors. The chemoattractant receptors on the surface of leukocytes then bind to chemotactic cytokines secreted by the cells at the site of injury or infection. Receptor binding activates leukocytes, increases the adhesion of adhesion molecules that mediate transendothelial migration, and promotes directed migration of cells toward the source of the chemoattractant cytokine.

Chemotactic cytokines (leukocyte chemotactic/activating factors), also known as chemokines and also known as intercrine and SIS cytokines, are a group of inflammatory/immunoregulatory polypeptide factors with molecular weights of 6-15kDa that are released at the inflammatory site by a variety of cells, such as macrophages, monocytes, eosinophils, neutrophils, fibroblasts, vascular endothelial cells, smooth muscle cells and mast cells (Luster, New Eng. J. Med., 338, 436-. Furthermore, chemokines have been described in the following documents: oppenheim, j.j.et al, annu.rev.immunol., 9: 617 and 648 (1991); schall and Bacon, curr, opin, immunol, 6: 865-873 (1994); baggiolini, m., et al, and adv.immunol., 55: 97-179(1994). Chemokines have the ability to stimulate the migration of committed cells, a process known as chemotaxis. Each chemokine contains four cysteine residues (C) and two internal disulfide bonds. Chemokines can be divided into two subfamilies, based on whether the two amino-terminal cysteine residues are immediately adjacent (CC family) or separated by one amino acid (CXC family). These differences correlate with the fact that the two subfamilies constitute separate gene clusters. Within each gene cluster, chemokines typically show 25 to 60% sequence similarity. CXC chemokines, such as interleukin-8 (IL-8), neutrophil-activating protein-2 (NAP-2) and melanoma growth stimulating activity protein (MGSA), are chemotactic mainly for neutrophils and T lymphocytes, while CC chemokines, such as RANTES, MIP-1 α, MIP-1 β, monocyte chemotactic proteins (MCP-1, MCP-2, MCP-3, MCP-4 and MCP-5) and eotaxins (-1 and-2), are chemotactic for various cell types, especially macrophages, T lymphocytes, eosinophils, dendritic cells and basophils. The chemokines lymphotactin-1, lymphotactin-2 (all C chemokines) and fractalkine (CXXXC chemokine) also exist and do not belong to any of the major chemokine subfamilies described above.

MCP-1 (also known as MCAF (abbreviation for macrophage chemotaxis and activator) or JE) is a CC chemokine produced by monocytes/macrophages, smooth muscle cells, fibroblasts and vascular endothelial cells, leading to cell migration and cell adhesion of: mononuclear cells (see, e.g., Valente, A.J., et al., Biochemistry, 1988, 27, 4162; Matsushima, K., et al., J.Exp.Med., 1989, 169, 1485; Yoshimura, T., et al., J.Immunol., 1989, 142, 1956; Rollins, B.J., et al., Proc.Natl.Acad.Sci.USA, 1988, 85, 3738; Rollins, B.J., et al., Blood, 1991, 78, 1112; Jiang, Y., J.Immunol., 1992, 148, 2423; Vaddi, K., et al, J.Immunol., 153, 4721), memory T cells (see, e.g., Carr, M.J., et al., W.J.J., Ed., USA, J.S.R., USA, J.S.23, J.S.S.S.S.S.S.S.S.S. Ab., J., et al., USA, J., Ab., J., et al., Ab., J., Ab., FAS., J., Ab., J., et al., J., Ab., J., Ab., J., Ab., D.S. 11, Ab., J., Ab., D.S. Ab., D., D.S. No. Ab., D.S. 11, Ab., D, s.c., et al, j.exp.med., 1992, 175, 1271; kuna, p., et al, j.exp.med., 1992, 175, 489). In addition, high expression of MCP-1 has been reported in certain diseases where monocyte/macrophage and/or T cell accumulation is considered important in the initiation or progression of the disease, such as atherosclerosis (see, e.g., Hayes, I.M., et al, Arterioscler, Thromb, Vasc, biol, 1998, 18, 397; Takeya, M., et al, hum, Pathol, 1993, 24, 534; Yla-Herttuala, S., et al, Proc. Natl. Acad. Sci.USA, 1991, 88, 5252; Nelken, N.A., J.Clin. Invest, 1991, 88, 1121), rheumatoid Arthritis (see, e.g., Koch, A.E., J.Clin.EST.1992, 90, Akahoshi, 1991, J.Clin.S., 1121, Ak., Aust, Clin., Kl, 90, Ak. Ab, Ak, J.E., J.Clin. Clin. 35, 1997, Lo. mu. E., Kl, Kl.S., LR.S., R.S., 73, R.D.S., LR. E., LR., 73, LR., 51, 155), renal disease (see, e.g., Saitoh, a., et al., j.clin.lab.anal., 1998, 12, 1; yokoyama, h., et al, j.leukcc.biol., 1998, 63, 493), pulmonary fibrosis, pulmonary sarcoidosis (see, e.g., Sugiyama, y., et al, Internal Medicine, 1997, 36, 856), asthma (see, e.g., Karina, m., et al, j.invest.allergol.clin.immunol., 1997, 7, 254; stephene, t.h., am.j.respir.crit.careed, 1997, 156, 1377; sousa, a.r., et al., am.j.respir.cell mol.biol., 1994, 10, 142), multiple sclerosis (see, e.g., McManus, c.et al., j.neurommunol., 1998, 86, 20), psoriasis (see, e.g., gillitizer, r., et al., j.invest.dermotol., 1993, 101, 127), inflammatory bowel disease (see, e.g., Grimm, m.c., et al., j.leukoc.biol., 1996, 59, 804; reinecker, h.c., et al, Gastroenterology, 1995, 106, 40), myocarditis (see, e.g., Seino, y, et al, cytokines, 1995, 7, 301), endometriosis (see, e.g., Jolicoeur, c, et al, am.j.pathol., 1998, 152, 125), intraperitoneal adhesions (see, e.g., Zeyneloglu, h.b., et al, Human Reproduction, 1998, 13, 1194), congestive heart failure (see, e.g., aurus, p.et al, Circulation, 1998, 97, 1136), chronic liver disease (see, e.g., Marra, f.et al, am.j.pathol., 1998, 152, 423), viral meningitis (see, e.g., Lahrtz, f.et al, euro.j.27, msy, 1997, wasaki, 84; et al, j.rheumatol, 1997, 24, 1179) and sepsis (see, e.g., Salkowski, c.a.; et al, perfect. immun., 1998, 66, 3569). In addition, anti-MCP-1 antibodies have been reported to show inhibitory or therapeutic effects in animal models of the following diseases: rheumatoid arthritis (see, e.g., Schimmer, r.c., et al., j.immunol., 1998, 160, 1466; Schrier, d.j., j.leukoc.biol., 1998, 63, 359; Ogata, h., et al., j.pathol., 1997, 182, 106), multiple sclerosis (see, e.g., Karpus, w.j., et al., j.leukoc.biol., 1997, 62, 681), nephritis (see, e.g., Lloyd, c.m., et al, j.exp.d., 1997, 185, 1; Wada, t., FASEB j., 1996, 10, 1418), asthma (see, e.g., Gonza, j., a., J., J.exp. ep., J.D., 1997, 1998, 188, Lukaka, N.J., 1996, 10, 1418), atherosclerosis (see, e.g., J.A., J.120, J.J.J.J., J.J.22, J.J.J.J.J.22, D., 1998, J.J.J.J.D., 1998, J.J.J., 92, J.D., 92, J.J.D., 1998, J.J.D., 2000, J.D.I., 1998, J.D., 2000, J., 2000, J.D.I., 1998, J., 2000, J., 2000, D., J., I., 2000, J., 2000, J., 2000, D., 2000, D., I., 2000, J., h.b., et al, am.j.obstet.gynecol, 1998, 179, 438). Peptide antagonists of MCP-1, MCP-1(9-76), have also been reported to inhibit arthritis in mouse models (Gong, j. -h., j.exp., 4ed., 1997, 186, 131), studies in MCP-1-deficient mice have shown that MCP-1 is essential for monocyte recruitment in vivo (Lu, b., et al., j.exp.med., 1998, 187, 601; Gu, l., et al., mol.cell, 1998, 2, 275).

The published literature suggests that chemokines, such as MCP-1 and MIP-1 alpha, attract monocytes and lymphocytes to the site of disease, mediate their activation, and are therefore thought to be intimately involved in the initiation, progression and maintenance of diseases that involve deep monocytes and lymphocytes, such as atherosclerosis, restenosis, rheumatoid arthritis, psoriasis, asthma, ulcerative colitis, nephritis (nephropathy), multiple sclerosis, pulmonary fibrosis, myocarditis, hepatitis, pancreatitis, sarcoidosis, Crohn's disease, endometriosis, congestive heart failure, viral meningitis, cerebral infarction, neuropathy, Kawasaki disease and sepsis (see, e.g., Rovin, B.H., et al, am. J.Kidney. Dis., 1998, 31, 1065; Lloyd, C.et al., Curra. Opin. Nephrol. hypertens, 1998, 7, 281; Conti, P.p., early, Prolec, et al, Inc., in the same, et al, and the like, 1998, 19, 121; ransohoff, r.m., et al, trends neurosci, 1998, 21, 154; MacDermott, r.p., et al, infilamatorybowel Diseases, 1998, 4, 54).

Chemokines bind to specific cell surface receptors belonging to the family of G-protein-coupled seven transmembrane domain proteins (Horuk, Trends pharm. sci., 15, 159-165(1994)), and they are known as "chemokine receptors". Upon binding their cognate ligands, chemokine receptors transduce intracellular signals through the associated trimeric G proteins, resulting in rapid increases in intracellular calcium concentrations, changes in cell shape, increased expression of cell adhesion molecules, degranulation, and promotion of cell migration, among other responses.

Genes encoding specific chemokine receptors have been cloned and these receptors are now known as G-protein-coupled seven transmembrane receptors present on a variety of leukocyte populations. To date, at least five CXC chemokine receptors (CXCR1-CXCR5) and eight CC chemokine receptors (CCR1-CCR8) have been identified. For example, IL-8 is a ligand of CXCR1 and CXCR2, MIP-1 α is a ligand of CCR1 and CCR5, MCP-1 is a ligand of CCR2A and CCR2B (for reference, see, for example, Holmes, W.E., et al, Science 1991, 253, 1278-. Pulmonary inflammation and granuloma formation have been reported to be inhibited in CCR 1-deficient mice (Gao, j. -l., et., j. exp. med., 1997, 185, 1959; Gerard, c., et al., j. clin.invest., 1997, 100, 2022), macrophage recruitment and atherosclerotic lesion formation were reduced in CCR 2-deficient mice (borring, l., et al., Nature, 1998, 394, 894; Kuziel, w.a., et al., proc.natl.acad.sci., USA, 1997, 94, 12053; Kurihara, t., et al, j. exp. med., 1997, 186, 1757; borring, l., et al, j. clin.ests, 1997, 100, 2552).

Thus, drugs that inhibit the binding of chemokines, such as MCP-1 and/or MIP-1 α, to these receptors, such as chemokine receptor antagonists, may be used as pharmaceutical agents to inhibit the effect of chemokines, such as MCP-1 and/or MIP-1 α, on target cells, but there is no prior art report that 3-cycloalkylaminopyrrolidine derivatives have such pharmacological effects. The identification of compounds that modulate CCR2 and/or CCR5 function represents an excellent drug design approach for the development of pharmacological agents for the treatment of inflammatory conditions and diseases associated with CCR2 and/or CCR5 activation, such as rheumatoid arthritis, lupus and other inflammatory diseases. The present invention provides a solution to the long-felt need in the field of chemokine receptor modulators and antagonists.

Disclosure of Invention

Object of the Invention

In view of the foregoing, it is an object of the present invention to provide chemokine receptor antagonists and chemokine receptor modulators for the treatment of rheumatoid arthritis.

It is another principal object of the present invention to provide chemokine receptor antagonists and their use as pharmaceutical ingredients.

It is a further object of the present invention to provide chemokine receptor modulators and their use as pharmaceutical ingredients.

It is a further object of the present invention to provide 3-cycloalkylaminopyrrolidine derivatives.

Another object of the invention relates to novel compounds and medical methods for treating inflammation.

It is a still further object of the present invention to provide novel compounds and pharmaceutical compositions thereof having anti-inflammatory and immunomodulatory biological activity that act via antagonism of the CCR2 receptor.

It is a further object of the present invention to provide 3-cycloalkylaminopyrrolidine derivatives and their use as modulators of chemokine receptors.

It is a further object of the present invention to provide 3-cycloalkylaminopyrrolidine derivatives and their use in the treatment and prevention of atherosclerosis and restenosis.

It is a further object of the present invention to provide 3-cycloalkylaminopyrrolidine derivatives and their use as modulators of the CCR5 receptor.

It is another principal object of the present invention to provide biologically active compounds of 3-cycloalkylaminopyrrolidine and pharmaceutical compositions thereof which act via antagonism of the CCR5 receptor.

Other objects and embodiments of the present invention will be discussed below. It is important to note, however, that many other embodiments of the present invention not described in this specification can still fall within the spirit and scope of the present invention and/or the claims.

Summary of The Invention

The present invention relates to compounds of formulae I and II:

or enantiomers, diastereomers, enantiomerically enriched mixtures, racemic mixtures thereof, prodrugs, crystalline forms, non-crystalline forms, amorphous forms thereof, solvates thereof, metabolites thereof, and pharmaceutically acceptable salts thereof, wherein the compositional variables are as provided herein.

The invention also relates to pharmaceutical compositions comprising anti-inflammatory and/or immunomodulatory compounds of formulae I and II as shown above, which act via antagonism of the CCR2 receptor (also known as MCP-1 receptor) and thus inhibit monocyte chemoattractant protein-1 (MCP-1).

The invention also relates to pharmaceutical compositions comprising anti-inflammatory and/or immunomodulatory compounds of formulae I and II as shown above, which act via antagonism of the CCR5 receptor (also known as MCP-1 receptor) and thus inhibit monocyte chemoattractant protein-1 (MCP-1).

The invention also relates to compounds of formulae I and II which are modulators of CCR2 chemokine receptor function and are useful in the prevention or treatment of inflammatory conditions and diseases such as rheumatoid arthritis, allergic diseases, psoriasis, atopic dermatitis, lupus and asthma.

The invention also relates to compounds of formulae I and II which are modulators of CCR5 chemokine receptor function and are useful in the prevention or treatment of inflammatory conditions and diseases such as rheumatoid arthritis, allergic diseases, psoriasis, atopic dermatitis, lupus and asthma.

The invention also provides pharmaceutical compositions comprising a compound selected from formulas I and II and the use of these compounds and compositions in the prevention or treatment of diseases in which the CCR2 chemokine receptor is involved.

The invention further provides pharmaceutical compositions comprising a compound selected from formulae I and II and the use of these compounds and compositions in the prevention or treatment of diseases in which the CCR5 chemokine receptor is involved.

The present invention further provides a method of treating inflammation, rheumatoid arthritis, lupus, systemic lupus erythematosus, atherosclerosis, restenosis, immune disorders, and transplant rejection in a mammal in need thereof, comprising administering to such mammal a therapeutically effective amount of a pharmaceutical composition comprising a compound according to formulas I and II in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.

The invention further provides compositions comprising a compound of the invention and a pharmaceutically acceptable carrier.

The invention further provides methods of modulating the activity of a chemokine receptor comprising contacting the chemokine receptor with a compound of the invention.

The invention further provides a method of treating a disease associated with chemokine receptor expression or activity in a patient comprising administering to the patient a therapeutically effective amount of a compound of the invention.

The invention further provides compounds of formulae I and II for use in therapy.

The invention further provides the use of a compound of formula I or II for the manufacture of a medicament for the treatment of a disease associated with chemokine receptor expression or activity.

Detailed Description

The present invention relates to compounds of formula I:

including enantiomers, diastereomers, enantiomerically enriched mixtures, racemic mixtures thereof, prodrugs, crystalline forms, non-crystalline forms, amorphous forms thereof, solvates thereof, metabolites thereof, and pharmaceutically acceptable salts thereof, wherein:

x is selected from the group consisting of a bond, aryl, mono-or poly-substituted aryl, heterocycle, mono-or poly-substituted heterocycle, heteroaryl, mono-or poly-substituted heteroaryl, carbocycle, mono-or poly-substituted carbocycle, and (CR)⁸R⁹)_nWherein n is 0-5;

y is a bond or is selected from the group consisting of oxygen, sulfur, nitrogen, amide bond, thioamide bond, sulfonamide, ketone, -CHOH-, -CHO-alkyl-, -alkyl-O-alkyl, oxime, and urea;

z is selected from the group consisting of carbocycle, aryl, heterocycle and heteroaryl, each having 0-3R¹⁰A substituent group, wherein R¹⁰Independently selected from the group consisting of: halogen, alkyl, alkenyl, alkynyl, alkoxy, cyclic alkoxy, heterocycloalkoxy, alkoxyalkyl, cyclic alkoxyalkyl, heterocycloalkoxyalkyl, alkylthioalkyl, cyclic alkylthioalkyl, heterocycloalkylthioalkyl, thioalkyl, mono-, di-or trihaloalkyl, mono-, di-or trihaloalkoxy, nitro, amino, mono-or di-substituted aminoalkyl, carboxyl, esterified carboxyl, acylamino, mono-or di-substituted acylamino, carbamate, mono-or di-substituted carbamate, sulfonamide, mono-or di-substituted sulfonamide, alkylsulfonyl, cyclic alkylsulfonyl, heterocycloalkylsulfonyl, arylsulfonyl, heteroarylsulfonyl, alkylsulfonyl, heterocycloalkylsulfonyl, arylsulfonyl, heteroarylsulfonyl, alkylsulfonyl, alkylcarbonyl, cycloalkylcarbonyl, heterocycloalkylcarbonyl, arylcarbonyl, heteroarylcarbonyl, thioamido, cyano, R^10a-carbocyclic ring, R^10a-heterocycle, R^10a-aryl and R^10a-heteroaryl, wherein R is^10aIs H, halogen, OH, amino, mono-or di-substituted amino, mono-, di-or tri-haloalkyl, alkoxyA group, mono-, di-or tri-haloalkoxy, amide, sulfonamide, carbamate, urea or cyano;

R¹independently selected from the group consisting of: carbocycle, heterocycle, aryl, heteroaryl, arylalkyl, heteroarylalkyl, arylalkenyl, heteroarylalkenyl, arylalkynyl, heteroarylalkynyl, arylaminocarbonyl, heteroarylaminocarbonyl, arylamido, heteroarylamido, arylureido, heteroarylureido, aryloxy, heteroaryloxy, arylalkoxy, heteroarylalkoxy, arylamino and heteroarylamino, wherein the carbocycle, heterocycle, aryl or heteroaryl is substituted with 0-3R^1aIs substituted in which R^1aIndependently selected from the group consisting of: halogen, alkyl, alkenyl, alkynyl, alkoxy, cyclic alkoxy, heterocycloalkoxy, alkoxyalkyl, cyclic alkoxyalkyl, heterocycloalkoxyalkyl, alkylthioalkyl, cyclic alkylthioalkyl, heterocycloalkylthioalkyl, hydroxyalkyl, mono-, di-or trihaloalkyl, mono-, di-or trihaloalkoxy, nitro, amino, mono-or di-substituted aminoalkyl, aminocarbonyl, mono-or di-substituted aminocarbonyl, cyclic aminocarbonyl, aminosulfonyl, mono-or di-substituted aminosulfonyl, alkylcarbonyl, cyclic alkylcarbonyl, heterocycloalkylcarbonyl, arylcarbonyl, heteroarylcarbonyl, formyl, alkylsulfonyl, cyclic alkylsulfonyl, heterocycloalkylsulfonyl, heteroaryl, alkoxy, haloalkoxy, haloalkylthio, amino, mono-or di-substituted amino, alkoxycarbonyl, heterocycloalkylthioalkyl, amino, mono-or di-substituted amino, alkylcarbonyl, Arylsulfonyl, heteroarylsulfonyl, carboxylic acid, esterified carboxylic acid, alkylcarbonamido, cyclic alkylcarbonamido, heterocycloalkylcarbonamido, arylcarbonylamino, heteroarylcarbonylamino, cyano, arylalkyl, heteroarylalkyl, aryloxyalkyl, heteroaryloxyalkyl, arylthioalkyl, heteroarylthioalkyl, carbamate, mono-or di-substituted carbamate, R^1b-carbocyclic ring, R^1b-heterocycle, R^1b-aryl and R^1b-heteroaryl, wherein R is^1bIs H, halogen, OH, amino, mono-or di-substituted amino, mono-, di-or tri-haloalkyl, alkoxy, mono-, di-or tri-haloalkoxy, hydroxyalkyl, alkoxyalkyl, aminoAlkyl, mono-or di-substituted aminoalkyl, amide, sulfonamide, carbamate, urea, or cyano;

R²independently selected from the group consisting of: H. amino, mono-or di-substituted amino, OH, carboxyl, esterified carboxyl, amide, N-monosubstituted amide with N, N-disubstituted amide, cyano, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, alkoxy, thioalkyl, mono-, di-or trihaloalkyl, halogen, aryl and heteroaryl;

optionally R¹And R²Can be bonded to each other to form a spiro ring;

R³and R⁴Independently selected from the group consisting of: H. amino, OH, alkyl, haloalkyl, dihaloalkyl, trihaloalkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, alkoxy, and thioalkyl;

optionally R³And R⁴May occupy multiple positions in the cycloalkyl ring;

optionally R¹And R³Can be cyclized to form a cyclic structure having 0 to 3R^aCarbocyclic or heterocyclic ring of substituents wherein R^aSelected from the group consisting of halogen, alkyl, alkoxy, thioalkyl, mono-, di-or trihaloalkyl, mono-, di-or trihaloalkoxy, nitro, amino, carboxyl, esterified carboxyl, amido, thioamido, cyano, mono-, di-or poly-substituted aryl or mono-, di-or poly-substituted heterocycle, optionally wherein said substituted aryl and substituted heterocycle are substituted with 0-3R^bIs substituted in which R^bSelected from the group consisting of halogen, alkyl, alkoxy, thioalkyl, mono-, di-or trihaloalkyl, mono-, di-or trihaloalkoxy, nitro, amino, carboxyl, esterified carboxyl, amido, thioamido and cyano;

optionally R³And R⁴Can be cyclized to form a bridged bicyclic system having methylene or ethylene groups or selected fromN, O and S;

optionally R³And R⁴Can be cyclized to form a spiro ring;

R⁵independently selected from the group consisting of hydrogen, alkyl, and formyl; and when R is⁵When alkyl, the nitrogen may optionally be in the form of an N-oxide;

R⁶and R⁷Each independently selected from the group consisting of: h; c₁-C₁₀Alkyl radical, wherein said C₁-C₁₀The alkyl group may optionally be interrupted by oxygen (O), Nitrogen (NH), or sulfur (S); a carbocyclic ring; a heterocycle; an alkoxy group; a cycloalkoxy group; a heterocyclic alkoxy group; mono-, di-or tri-haloalkyl; mono-, di-or tri-haloalkoxy; an aryloxy group; a heteroaryloxy group; an arylalkoxy group; heteroarylalkoxy; an aryloxyalkyl group; heteroaryloxyalkyl; an arylalkoxyalkyl group; a heteroaryl alkoxyalkyl group; an aryl group; a heteroaryl group; an arylalkyl group; a heteroarylalkyl group; a hydroxyalkyl group; an alkoxyalkyl group; a cycloalkoxyalkyl group; a heterocycloalkoxyalkyl group; an aminoalkyl group; mono-or di-substituted aminoalkyl; an arylaminoalkyl group; heteroarylaminoalkyl groups; an alkylthio alkyl group; cycloalkylthioalkyl; (ii) heterocycloalkylthioalkyl; arylthioalkyl; heteroarylthioalkyl; an alkylsulfonylalkyl group; a cycloalkylsulfonylalkyl group; heterocycloalkylsulfonylalkyl; arylsulfonylalkyl; heteroarylsulfonylalkyl; an aminocarbonyl group; mono-or di-substituted aminocarbonyl; aminocarbonylalkyl; mono-or di-substituted aminocarbonylalkyl; alkylcarbonylalkyl; a cycloalkylcarbonylalkyl group; heterocycloalkylcarbonylalkyl; an alkylcarbonamidoalkyl group; a cycloalkyl carbonylaminoalkyl group; a heterocycloalkyl carboamidoalkyl group; arylcarbonylaminoalkyl; heteroaryl carboxanylaminoalkyl; arylsulfonylaminoalkyl; and heteroarylsulfonylaminoalkyl;

alternatively, R⁶And R⁷May be cyclized to form a carbocyclic or heterocyclic ring, or a spiro or spiroheterocyclic ring;

R⁸and R⁹Independently selected from H, OH, ammoniaA group consisting of alkyl, arylalkyl, heteroarylalkyl, aryl, heteroaryl, alkoxy, alkenyl, alkynyl, alkoxyalkyl, mono-or di-substituted amino, carbocycle, and heterocycle;

optionally R⁸And R⁹May be cyclized to form a 3-7 membered carbocyclic or heterocyclic ring;

r＝0-3。

in a further embodiment, the invention relates to a compound of formula II:

including its enantiomers, diastereomers, enantiomerically enriched mixtures, racemic mixtures thereof, prodrugs, crystalline forms, non-crystalline forms, amorphous forms thereof, solvates thereof, metabolites thereof, and pharmaceutically acceptable salts, wherein the compositional variables are as provided above.

In some embodiments, X may be selected from aryl, mono-or poly-substituted aryl, heterocycle, heteroaryl, mono-or poly-substituted heteroaryl, carbocycle, mono-or poly-substituted carbocycle, and (CR)⁸R⁹)_nWhere n is 0-5 (e.g., n is 0, 1, 2, 3,4, or 5).

In some embodiments, X is a bond, heterocycle, mono-or poly-substituted heterocycle, heteroaryl, mono-or poly-substituted heteroaryl, or (CR)⁸R⁹)_nWherein n is 0 to 3.

In some embodiments, X is a heterocycle, a mono-or poly-substituted heterocycle, a heteroaryl, or a mono-or poly-substituted heteroaryl.

In some embodiments, X is (CR)⁸R⁹)_nWherein n is 0 to 3.

In some embodiments, X is CH₂。

In some embodiments, Y is a bond or-alkyl-O-alkyl-.

In some embodiments, -X-Y-is- (CR)⁸R⁹)_n-NH-CO-, -alkyl-O-alkyl-, heterocycle, or heteroaryl.

In some embodiments, -X-Y-is-CH₂-NH-CO-、-CH₂-O-CH₂-, azetidine, pyrrolidine, piperidine, imidazole or 4, 5-dihydroisoAnd (3) azole.

In some embodiments, -X-Y-is-CH₂-NH-CO-。

In some embodiments, Z is aryl or heteroaryl, each substituted with 0-3R¹⁰And (4) substituent substitution.

In some embodiments, Z is 6-membered aryl or 6-membered heteroaryl, each substituted with 0-3R¹⁰And (4) substituent substitution.

In some embodiments, Z is phenyl, pyridyl or pyrimidinyl, each substituted with 0-3R¹⁰And (4) substituent substitution.

In some embodiments, Z is phenyl, pyridyl, or pyrimidinyl, each substituted with at least one mono-, di-, or tri-haloalkyl.

In some embodiments, Z is:

in some embodiments, Z is:

in some embodiments, R¹The carbocyclic substituents of (A) are intended to include, for example, cycloalkyl groups of 3 to 10 carbon atoms and bicyclic and polycyclic bridging systems, e.g. norbornyl, adamantyl and bicyclo [2.2.2]And (4) octyl. R¹The carbocycle of (a) may also be further substituted with a heterocyclic or heteroaryl ring, such as pyridyl, pyrrolidinyl and all those defined under X above.

R¹Specific examples of the substituent include phenyl, pyridin-2-yl, 4-methylphenyl, 3-methyl-phenyl, 2-methylphenyl, 4-bromophenyl, 3-bromophenyl, 4-chlorophenyl, 3-chloro-phenyl, 4-trifluoromethylphenyl, 3-trifluoromethylphenyl, 2-methoxyphenyl, 3-pyridyl, 4-pyridyl, 2-methoxy-5-pyridyl, 2-ethoxy-5-pyridyl, 3, 4-methylenedioxyphenyl, 4-fluorophenyl, 3-trifluoromethyl-1H-pyrazol-1-yl, 3-fluorophenyl, 4-methoxyphenyl, 3-methoxyphenyl, Pyridin-4-yl, pyridin-3-yl, 5-methylpyridin-2-yl, 6-methylpyridin-2-yl, quinolin-4-yl, 3-methyl-1H-pyrazol-1-yl, 3, 5-dimethyl-1H-pyrazol-1-yl, 4-trifluoromethylphenyl, 3, 4-methylenedioxyphenyl, 4-cyanophenyl, 4- (methylaminocarbonyl) phenyl, 1-oxido (oxido) pyridin-4-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, 4-methylpyridin-2-yl, 5-methyl-pyridin-2-yl, 5-methylpyridin-2-yl, and mixtures thereof, 6-methylpyridin-2-yl, 6-methoxypyridin-3-yl, 6-methylpyridin-3-yl, 6-ethylpyridin-3-yl, 6-isopropylpyridin-3-yl, 6-cyclopropylpyridin-3-yl, 1-oxidopyridin-2-yl, 3-cyanophenyl, 3- (methylaminocarbonyl) -phenyl, 4- (morpholin-4-ylcarbonyl) -phenyl, 5- (morpholin-4-ylcarbonyl) pyridin-2-yl, 6- (morpholin-4-ylcarbonyl) pyridin-3-yl, methyl-amino-3-yl, methyl-pyridin-3-yl, methyl-pyridin-3-, 4- (4-methylpiperazin-1-yl-carbonyl) phenyl, 6- (azetidin-1-yl) pyridin-3-yl, 5-cyanopyridin-2-yl, 6-cyanopyridin-3-yl, 5- (methoxy-methyl) pyridin-2-yl, 5- (1-hydroxy-1-methylethyl) pyridin-2-yl, 5-dimethylaminomethyl, 4-ethylaminocarbonylphenyl, 4-isopropylaminocarbonylphenyl, 4-tert-butylamino-carbonylphenyl, 4-dimethylaminocarbonylphenyl, 4- (azetidin-1-yl) carbonylphenyl, 4- (pyrrolidin-1-yl) carbonylphenyl, methyl-ethyl-pyridin-2-yl, methyl-ethyl-4-yl, methyl-4-carbonylamino-phenyl, methyl-4-ethyl-4-carbonylamino-phenyl, methyl-4-isopropylaminocarbonyl-phenyl, 4- (do-Lin-4-yl) carbonylphenyl, 4- (dimethyl-aminocarbonyl) -2-methylphenyl, 2-methyl-4- (methylamino-carbonyl) phenyl, 3-methyl-4- (methylaminocarbonyl) phenyl, 4- (dimethylaminocarbonyl) -3-methylphenyl, 3-methyl-4- (pyrrolidin-1-ylcarbonyl) phenyl, 4- (dimethylaminocarbonyl) -3-fluorophenyl, 4- [ (2, 2, 2-trifluoroethyl) aminocarbonyl]Phenyl, 3-fluoro-4-methylaminocarbonyl-phenyl, 4-ethyl-aminocarbonyl-3-fluorophenyl, 3-methylaminocarbonylphenyl, 3-dimethyl-aminocarbonylphenyl, 5-dimethylaminocarbonyl-2-methoxyphenyl, 2-methoxy-5-methyl-aminocarbonylphenyl, 3- (methylaminocarbonamido) phenyl, 6- (morpholin-4-yl) -pyridin-3-yl, 6-dimethylaminopyridin-3-yl, 6-isopropylaminopyridin-3-yl, 6- (pyrrolidin-1-yl) pyridin-3-yl, 6-cyclopropylaminopyridin-3-yl, p-ropylamino-pyridin-3-yl, p-propylamino-phenyl, p-propylamino-pyridin-3-yl, p-propylamino-phenyl, p-propylamino-pyridin-, 6-ethoxypyridin-3-yl, 6- (2-fluoroethoxy) pyridin-3-yl, 6- (2, 2-difluoroethoxy) pyridin-3-yl, 6- (2, 2, 2-trifluoroethoxy) -pyridin-3-yl, 4-iodophenyl, 5- (pyrrolidin-1-ylcarbonyl) -2-pyridyl, 5- (morpholin-4-yl-carbonyl) -2-pyridyl, 5-dimethylaminocarbonyl-2-pyridyl, 4-methylaminocarbonyl-aminophenyl, 6- (1-hydroxy-1-methylethyl) pyridin-3-yl, 4- (1-hydroxy-1-methylethyl) -phenyl, methyl-ethyl, ethyl-phenyl, methyl-phenyl, ethyl, methyl-phenyl, 4- (methoxymethyl) phenyl, 3-fluoro-4- (methoxymethyl) phenyl, 4- (dimethyl-amino) phenyl, 4- (dimethylamino) -3-fluorophenyl, 1H-indazol-5-yl, 1-methyl-1H-indazol-5-yl, 2-methyl-1H-indazol-5-yl, 1, 3-thiazol-2-yl, 5-ethyl-1, 3-thiazol-2-yl, 5- (methyl-aminocarbonyl) -1, 3-thiazol-2-yl, 1, 3-thiazol-5-yl, 2- (methoxycarbonylamino) -1, 3-thiazol-5-yl, methyl-ethyl-1, 3-thiazol-5-yl, methyl, 2-isopropyl-1, 3-thiazol-5-yl, 5- (pyridin-3-yl) -1, 3-thiazol-2-yl, 5- (morpholin-4-ylcarbonyl) -1, 3-thiazol-2-yl, 5-aminocarbonyl-1, 3-thiazol-2-yl, 5-dimethylaminocarbonyl-1, 3-thiazol-2-yl, 5- (pyrrolidin-1-ylcarbonyl) -1, 3-thiazol-2-yl, 5-allyl-1, 3-thiazol-2-yl, 5-propyl-1, 3-thiazol-2-yl, 5-ethylaminocarbonyl-1, 3-thiazol-2-yl, 5-phenyl-1, 3-thiazol-2-yl, 5-methyl-1, 3-thiazol-2-yl, 5-hydroxymethyl-1, 3-thiazol-2-yl, 5- (1-hydroxy-1-methylethyl) -1, 3-thiazol-2-yl, 5-methoxy-methyl-1, 3-thiazol-2-yl, 5- (2-pyridyl) -1, 3-thiazol-2-yl, 2- (pir-ethyl)Pyrrolidin-1-yl) -1, 3-thiazol-4-yl, 2- (morpholin-4-yl) -1, 3-thiazol-4-yl, 2-methyl-1, 3-thiazol-5-yl, 2- (1-hydroxy-1-methylethyl) -1, 3-thiazol-5-yl, 2- (pyrrolidin-1-yl) -1, 3-thiazol-5-yl, 2-ethoxy-1, 3-thiazol-5-yl, 2-ethyl-1, 3-thiazol-5-yl, 2- (pyrrolidin-1-ylmethyl) -1, 3-thiazol-5-yl, methyl-1, 3-thiazol-5-yl, methyl-1, 3-ethyl-5-yl, methyl-1, 3-methyl-thiazol-5-yl, methyl-1, 3-ethyl-5-, 2- (morpholin-4-yl) -1, 3-thiazol-5-yl, 2-methoxy-methyl-1, 3-thiazol-5-yl, 2-isobutyl-1, 3-thiazol-5-yl, 2-ethylaminocarbonyl-1, 3-thiazol-5-yl, 2- (pyrrolidin-1-ylcarbonyl) -1, 3-thiazol-5-yl, 2- (morpholin-4-ylcarbonyl) -1, 3-thiazol-5-yl, 2- (3-pyridyl) -1, 3-thiazol-5-yl, 2- (2-pyridyl) -1, 3-thiazol-5-yl, 2- (morpholin-4-ylcarbonyl) -1, 3-thiazol-5-yl, 2- (2-pyridyl) -1, 3-thiazol, 4-methyl-1, 3-thiazol-2-yl, 1, 3-benzo-thiazol-2-yl, pyrimidin-5-yl, pyrimidin-2-yl, pyridazin-4-yl, pyridazin-3-yl, pyrazin-2-yl, 2-methoxypyrimidin-5-yl, 2-ethoxypyrimidin-5-yl, 2- (2-fluoroethoxy) pyrimidin-5-yl, 2-methylpyrimidin-5-yl, 2-ethylpyrimidin-5-yl, 2-isopropylpyrimidin-5-yl, 2-cyclopropylpyrimidin-5-yl, pyrimidin-4-yl, 4- (pyrimidin-5-yl) phenyl, 4- (1,3-Oxazol-2-yl) phenyl, 4- (1H-imidazol-1-yl) phenyl, 4- (morpholin-4-yl) phenyl, 5- (pyrazin-2-yl) pyridin-2-yl, 4- (1-methyl-1H-imidazol-5-yl) phenyl, 4- (4, 6-dimethylpyrimidin-5-yl) phenyl, 6-bromopyridin-3-yl, 5-bromopyridin-2-yl, 4'- (methylsulfonyl) biphenyl-4-yl, 3' - (methoxy-carbonyl) -biphenyl-4-yl, 4- (2, 3-dihydro-1, 4-benzodiEn-6-yl) phenyl, 4' - (dimethyl-amino) -biphenyl-4-yl, 4- (pyridin-3-yl) phenyl, 4- (1H-pyrazol-4-yl) phenyl, 4- (3,3' -bipyridin-6-yl, 4- (3,4' -bipyridin-6-yl, 5- (3-acetylphenyl) pyridin-2-yl, 5- [3- (dimethyl-amino) phenyl]Pyridin-2-yl, 5- [3- (trifluoromethyl) phenyl]Pyridin-2-yl, 5- [4- (methyl-sulfonyl) phenyl]Pyridin-2-yl, 5- (4-methoxy-phenyl) pyridin-2-yl, 5- (3-methoxy-phenyl) -pyridin-2-yl, 5- [3- (aminocarbonyl) -phenyl]Pyridin-2-yl, 5- (4-fluoro-phenyl) pyridin-2-yl, 5- (3, 4-difluorophenyl) pyridin-2-yl, 5- (3, 5-dimethylisoOxazol-4-yl) pyridin-2-yl, 5- (1-methyl-1H-pyrazol-4-yl) pyridin-2-yl, 5- (1-benzofuran-2-yl) pyridin-2-yl, 5- (1, 3-benzodioxol-5-yl) pyridin-2-yl, 5- (2-formyl-phenyl) pyridin-2-yl, 4- (2' -formylbiphenyl-4-yl, 5- (1,3-Oxazol-2-yl) pyridin-2-yl, 6- (1,3-Oxazol-2-yl) pyridin-3-yl, 4- (1, 3-thiazol-2-yl) phenyl, 5- (1, 3-thiazol-2-yl) pyridin-2-yl, 6- (1, 3-thiazol-2-yl) pyridin-3-yl, 6- (1H-imidazol-1-yl) pyridin-3-yl]5- (1H-imidazol-1-yl) pyridin-2-yl, 6-phenylpyridin-3-yl, 5- (pyrimidin-5-yl) pyridin-2-yl, 5- (pyrimidin-2-yl) pyridin-2-yl, 5- (3-aminocarbonylphenyl) pyridin-2-yl, 4- (1-methyl-1H-imidazol-4-yl) phenyl, 4- (1H-imidazol-4-yl) phenyl]5- [2- (hydroxymethyl) phenyl]Pyridin-2-yl, 2' - (hydroxymethyl) biphenyl-4-yl, 5- {2- [ (dimethylamino) methyl]Phenyl } pyridin-2-yl, 2' - [ (dimethylamino) methyl]Biphenyl-4-yl, 5-fluoromethylpyrazin-2-yl, 5-difluoro-methyl-pyrazin-2-yl, 5-methylpyrazin-2-yl, 2-methyl-pyrimidin-5-yl, 2-fluoromethyl-pyrimidin-5-yl, 2-difluoromethylpyrimidin-5-yl, 2-trifluoro-methylpyrimidin-5-yl, 2-cyclopropylpyrimidin-5-yl, isothiazol-5-yl, 3-methylisothiazol-5-yl, 3-fluoromethyl-isothiazol-5-yl, 4- (dimethylamino-carbonyl) phenyl, 4- (methylaminocarbonyl) -phenyl, 5-difluoromethylpyrazin-2-yl, 5-difluoro-methyl-pyrazin-5-yl, 2-difluoromethylpyrimidin-5-yl, 2-trifluoromethyl-pyrimidin-5-yl, 2-, 4- (morpholin-4-ylcarbonyl) phenyl, 4- (piperidin-1-ylcarbonyl) phenyl, 3-fluoro-4- (pyrrolidin-1-ylcarbonyl) phenyl, 5- (pyrrolidin-1-yl-carbonyl) pyridin-2-yl, 5- (dimethyl-aminocarbonyl) pyridin-2-yl, 5- (morpholin-4-yl-carbonyl) -pyridin-2-yl, quinolin-4-yl, 6-methoxypyridin-3-yl, 6- (morpholin-4-yl) pyridin-3-yl, 4- (dimethyl-aminomethyl) phenyl, 5- (di-methyl) (di-n-ethyl) pyridin-2-ylDimethylaminomethyl) pyridin-2-yl, 5- (dimethyl-aminocarbonyl) -pyridin-2-yl, 4- [ hydroxy- (pyridin-3-yl) methyl]Phenyl, 6- [ (hydroxy- (pyridin-3-yl) methyl)]Pyridin-3-yl, 6- (dimethyl-aminocarbonyl) pyridin-3-yl, 4- (4-hydroxypiperidin-1-ylcarbonyl) phenyl, 4- (4-methoxy-piperidin-1-ylcarbonyl) phenyl, 5- (4-methoxypiperidin-1-ylcarbonyl) -pyridin-2-yl, 6- (4-methoxy-piperidin-1-ylcarbonyl) pyridin-3-yl, phenoxy, benzyloxy, 2-thienyl, 5- (methoxy-methyl) -1, 3-thiazol-2-yl, 5- (morpholin-4-ylcarbonyl) -1, 3-thiazol-2-yl, methyl-ethyl-phenyl, methyl-phenyl, ethyl-, 2-isopropyl-1, 3-thiazol-5-yl, 2- (methoxymethyl) -1, 3-thiazol-5-yl, 5- (methoxymethyl) -1, 3-thiazol-2-yl, 4- (pyrimidin-2-yl) phenyl, 4- (pyrimidin-4-yl) phenyl, and 5- (methoxymethyl) pyridin-2-yl.

In some embodiments, R¹Is aryl or heteroaryl, each substituted by 0-3R^1aAnd (4) substitution.

In some embodiments, R¹Is phenyl, pyridyl, pyrimidinyl, pyridazinyl or thiazolyl, each substituted by 0-3R^1aAnd (4) substitution.

In some embodiments, R¹Is aryl or heteroaryl, each substituted by 0-3R^1aAlkyl, alkoxy, alkoxyalkyl, hydroxyalkyl, mono-or di-substituted aminoalkyl, aminocarbonyl, mono-or di-substituted aminocarbonyl, cyclic aminocarbonyl, alkylcarbonyl, formyl, carboxylic acid, carbamate, mono-or di-substituted carbamate, R^1b-aryl or R^1b-heteroaryl substitution.

In some embodiments, R¹Is aryl or heteroaryl, each substituted by 0-1R^1b-aryl or R^1b-heteroaryl substitution.

In some embodiments, R¹Is aryl or heteroaryl, each being substituted by phenyl, pyridyl, pyrimidyl,Oxazolyl, thiazolyl, or imidazolyl.

In some embodiments, R¹Is substituted by phenyl, pyridyl, pyrimidyl,Heteroaryl substituted with oxazolyl, thiazolyl or imidazolyl.

In some embodiments, R²The group can be selected from H, amino, mono-or di-substituted amino, OH, carboxyl, esterified carboxyl, amide, N (C)₁-C₅) Mono-substituted amides with N (C)₁-C₅)、N(C₁-C₅) -disubstituted amides, cyano groups, (C)₁-C₈) Alkyl, (C)₂-C₈) -alkenyl, (C)₂-C₈) Alkynyl, (C)₅-C₇) -cycloalkyl, (C)₅-C₇) -cycloalkenyl, alkoxy, alkoxyalkyl, thioalkyl, mono-, di-or trihaloalkyl, halogen, aryl or heteroaryl.

In some embodiments, R²Is H or OH.

In some embodiments, R²Is OH.

In some embodiments, R¹Is aryl or heteroaryl, each substituted by 0-1R^1b-aryl or R^1b-heteroaryl substitution; r²Is OH.

In some embodiments, R³And R⁴The group substituents may be independently selected from the group consisting of: H. amino, OH, (C)₁-C₈) Alkyl, halo (C)₁-C₅) Alkyl, dihalo (C)₁-C₅) Alkyl, trihalo (C)₁-C₅) Alkyl, (C)₂-C₈) Alkenyl, (C)₂-C₈) Alkynyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, (C)₁-C₅) Alkoxy and thio (C)₁-C₅) An alkyl group.

In some instancesIn the embodiment, R³And R⁴Are all H.

In some embodiments, R⁵The substituents may be independently selected from hydrogen, (C)₁-C₈) Alkyl, formyl; and when R is⁵When alkyl, the nitrogen may optionally be in the form of an N-oxide.

In some embodiments, R⁵Is H.

In some embodiments, R⁶And R⁷Each substituent is independently selected from the group consisting of H, C₁-C₁₀Alkyl (optionally C)₁-C₁₀Alkyl may be interrupted by oxygen, nitrogen or sulfur), carbocycle, heterocycle, alkoxy, mono-, di-or tri-haloalkyl, mono-, di-or tri-haloalkoxy, cycloalkoxy, heterocycloalkoxy, aryloxy, heteroaryloxy, arylalkoxy, heteroarylalkoxy, aryloxyalkyl, heteroaryloxyalkyl, arylalkoxyalkyl or heteroarylalkoxyalkyl; aryl, heteroaryl, arylalkyl, heteroarylalkyl, hydroxyalkyl, alkoxyalkyl, cycloalkoxyalkyl, heterocycloalkoxyalkyl, aminoalkyl, mono-or di-substituted aminoalkyl, arylaminoalkyl, heteroarylaminoalkyl, alkylthioalkyl, cycloalkylthioalkyl, heterocycloalkylthioalkyl, arylthioalkyl, heteroarylthioalkyl (heteroarylthioalkyl), alkylsulfonylalkyl, cycloalkylsulfonylalkyl, heterocycloalkylsulfonylalkyl, arylsulfonylalkyl, heteroarylsulfonylalkyl, aminocarbonyl, mono-or di-substituted aminocarbonyl, aminocarbonylalkyl, mono-or di-substituted aminocarbonylalkyl, alkylcarbonylalkyl, cycloalkylcarbonylalkyl, heterocycloalkylcarbonylalkyl, alkylcarboxamidoalkyl, cycloalkylcarbonylaminoalkyl, heterocycloalkylcarbonylaminoalkyl, cycloalkylcarbonylaminoalkyl, heterocycloalkylalkyl, heterocycloalkylcarbonylaminoalkyl, heterocycloalk, Arylcarbonylaminoalkyl, heteroarylcarbonylaminoalkyl, arylsulfonylaminoalkyl and heteroarylsulfonylaminoalkyl groups. R⁶And R⁷Specific examples of substituents are as described above for R¹Those defined are the same.

In some embodiments of the present invention, the substrate is,R⁶and R⁷Independently selected from H, C₁-C₁₀Alkyl, hydroxyalkyl and alkoxyalkyl.

In some embodiments, R⁶And R⁷One is H and the other is H, C₁-C₁₀Alkyl, hydroxyalkyl or alkoxyalkyl.

In some embodiments, R⁶And R⁷Are all H.

In some embodiments, R⁸And R⁹The substituents are independently selected from the group consisting of H, OH, amino, (C)₁-C₈) Alkyl, arylalkyl, heteroarylalkyl, aryl, heteroaryl, (C)₁-C₈) -alkoxy, (C)₂-C₈) -alkenyl, (C)₂-C₈) -alkynyl, (C)₁-C₈) Alkoxyalkyl, mono (C)₁-C₈) -or di (C)₁-C₈) -substituted amino, carbocyclic and heterocyclic ring. When R is⁸And R⁹When cyclized to form a 3-to 7-membered carbocyclic or heterocyclic ring, such groups may be, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, iso-pentylAzolyl thiazolyl, dihydroOxazolyl, pyridyl, pyrimidinyl, or imidazolyl.

In some embodiments, R⁸And R⁹Are all H.

In some embodiments, r is 0, 1, 2, or 3. In a further embodiment, r is 1.

In some embodiments:

x is a bond, heterocycle, mono-or polysubstituted heterocycle, heteroaryl, mono-or polysubstituted heteroaryl or (CR)⁸R⁹)_nWherein n is 0-3;

y is a bond or-alkyl-O-alkyl-;

z is aryl or heteroaryl, each substituted by 0-3R¹⁰Substituent group substitution;

R¹is aryl or heteroaryl, each substituted by 0-3R^1aSubstitution;

R²is H or OH;

R³and R⁴Are all H;

R⁵is hydrogen, alkyl or formyl;

R⁶and R⁷Is H, C₁-C₁₀Alkyl, hydroxyalkyl or alkoxyalkyl;

R⁸and R⁹Are all H;

r is 1.

In some embodiments:

-X-Y-is- (CR)⁸R⁹)_n-NH-CO-, -alkyl-O-alkyl-, heterocycle or heteroaryl;

z is aryl or heteroaryl, each substituted by 0-3R¹⁰Substituent group substitution;

R¹is aryl or heteroaryl, each substituted by 0-3R^1aSubstitution;

R²is H or OH;

R³and R⁴Are all H;

R⁵is hydrogen;

R⁶and R⁷Are all H;

R⁸and R⁹Are all H;

r is 1.

In some embodiments:

-X-Y-is-CH₂-NH-CO-；

Z is phenyl, pyridyl or pyrimidinyl, each substituted with at least one mono-, di-or tri-haloalkyl;

R¹is aryl or heteroaryl, each being substituted by phenyl, pyridyl, pyrimidyl,Oxazolyl, thiazolyl, or imidazolyl;

R²is OH;

R³and R⁴Are all H;

R⁵is hydrogen;

R⁶and R⁷Are all H;

R⁸and R⁹Are all H;

r is 1.

In some embodiments:

-X-Y-is-CH₂-NH-CO-；

Z is phenyl substituted by at least one mono-, di-or tri-haloalkyl;

R¹is a pyridyl group, a pyrimidyl group,Heteroaryl substituted with oxazolyl, thiazolyl, or imidazolyl;

R²is OH;

R³and R⁴Are all H;

R⁵is hydrogen;

R⁶and R⁷Are all H;

R⁸and R⁹Are all H;

r is 1.

In various places in the specification, substituents for the compounds of the invention are disclosed in several groups or ranges. In particular, the invention includes various subcombinations of members of such groups and ranges. For example, the term "C_1-6Alkyl "specifically discloses individually methyl, ethyl, C₃Alkyl radical, C₄Alkyl radical, C₅Alkyl and C₆An alkyl group.

For compounds of the invention in which a variable occurs more than once, each variable may be a different moiety selected from the markush group defining the variable. For example, if a structure is described as having two R groups present on the same compound at the same time, the two R groups may represent different moieties selected from the markush group defining R.

It is further appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination.

The term aryl is intended to include aromatic carbocyclic groups such as phenyl, biphenyl, indenyl, naphthyl, as well as aromatic carbocyclic rings fused to a heterocycle such as benzothienyl, benzofuranyl, indolyl, quinolinyl, benzothiazolylOxazoles, benzimidazoles, isoquinolines, isoindolyl, benzotriazoles, indazoles, and acridinyl groups.

The term heteroaryl is intended to include mono-and polycyclic aromatic rings containing from 3 to 20, preferably from 4 to 10 ring atoms, at least one of which is a heteroatom, such as oxygen, sulfur, phosphorus or nitrogen. Examples of such groups include furyl, thienyl, pyrrolyl, imidazolyl, triazolylA group, a thiazolyl group, a tetrazolyl group,Azolyl radical, isoAzolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, quinolinyl, isoquinolinyl, quinoxalinyl, benzothiazolyl, benzolAzolyl, benzothienyl, or benzofuranyl.

The terms "cyclic alkyl", "cycloalkyl" and "carbocycle" are used interchangeably herein to refer to non-aromatic cyclic hydrocarbons (monocyclic and polycyclic), such as cyclized alkyl, alkenyl or alkynyl groups. In some embodiments, cycloalkyl is C_3-14、C_3-10、C_3-8、C_3-7、C_3-6Or C_3-5. In some embodiments, the cycloalkyl moieties each have 3 to 14, 3 to 10, or 3 to 7 ring-forming carbon atoms. In some embodiments, the cycloalkyl group has 0, 1, or 2 double or triple bonds. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl and the like. In this application, cycloalkyl is also intended to include bridged cyclic hydrocarbons such as adamantyl and the like.

Heterocycles are non-aromatic carbocyclic rings (monocyclic or polycyclic) that include one or more heteroatoms, such as nitrogen, oxygen, or sulfur, in the ring. In some embodiments, the ring can be ternary, quaternary, pentavalent, hexavalent, heptavalent, or octavalent. In some embodiments, the heterocyclic ring contains 1, 2, or 3 heteroatoms. The heterocyclic ring may be saturated or unsaturated. In some embodiments, the heterocyclic ring contains 0, 1 or 2 double or triple bonds. The ring-forming carbon atoms and heteroatoms may also carry oxo or sulfide substituents (e.g. CO, CS, SO)₂NO, etc.). Examples of the heterocyclic ring include tetrahydrofuryl, tetrahydrothienyl, morpholino, thiomorpholino, azetidinylPyrrolidinyl, piperazinyl, piperidinyl, pyranyl, and dinylAlkyl and thiazolidinyl.

In addition, when the heteroaryl or heterocyclic group is a nitrogen-containing heterocycle, the nitrogen may be modified to exist in the form of N → O (N-oxide), and such oxides are intended to be included within the scope of the present invention. In the case of sulfur-containing heterocycles, sulfur oxides are also intended to be included within the scope of the invention.

Monosubstituted aryl refers to an aryl group having one substituent. The polysubstituted aryl group means an aryl group having two or more substituents (for example, 2 to 4 substituents). Monosubstituted heteroaryl refers to heteroaryl having one substituent. Polysubstituted heteroaryl means heteroaryl having two or more substituents (e.g., 2 to 4 substituents). Monosubstituted cycloalkyl (or carbocycle) refers to a cycloalkyl group having one substituent. Polysubstituted cycloalkyl (or carbocycle) means cycloalkyl having two or more substituents, for example 2 to 4 substituents. Monosubstituted heterocyclic refers to a heterocyclic ring having one substituent. Polysubstituted heterocycle means a heterocycle having two or more substituents (e.g., 2 to 4 substituents).

The substituents on the aryl, arylalkyl, heteroaryl, heteroarylalkyl, carbocyclic (cycloalkyl) and heterocyclic groups of the present invention may be selected from the group consisting of halogen, alkyl, alkoxy, monohaloalkoxy, dihaloalkoxy, trihaloalkoxy, thioalkyl, monohaloalkyl, dihaloalkyl, trihaloalkyl, nitro, amino, carboxyl, esterified carboxyl, amide, thioamido and cyano. More precisely, the substituents may also be chosen from trifluoromethyl, C_1-4Alkyl, halo, trifluoromethoxy, fluoromethoxy, difluoromethoxy, C_1-5Alkoxy radical, C_1-5Alkanoyl radical, C_1-5Alkanoyloxy group, C_1-5Alkylamino radical, di (C)_1-5Alkyl) -amino, C_1-5Alkanoylamino, nitro, carboxy, carbamoyl, C_1-5Alkoxycarbonyl, mercapto, C_1-5Sulfonamido, carbamoyl C_1-5Alkyl, N- (C)_1-5Alkyl) carbamoyl radical C_1-5Alkyl, N- (C)_1-5Alkyl radical)₂Carbamoyl radical-C_1-5Alkyl, hydroxy C_1-5Alkyl and C_1-5Alkoxy radical C_1-4Alkyl groups.

The terms halo or halogen by themselves or as part of another substituent mean fluoro, chloro, bromo or iodo, unless otherwise specified. Similarly, terms such as haloalkyl are meant to include monohaloalkyl and polyhaloalkyl. For example, the term haloalkyl, such as halo (C)₁-C₄) Alkyl, is meant to include trifluoromethyl, 2, 2, 2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term alkyl, used alone or as a suffix, includes straight-chain and branched structures such as primary, secondary and tertiary alkyl groups. These groups may contain up to 15, preferably up to 8, more preferably up to 4 carbon atoms. In some embodiments, the alkyl group is C_1-10、C_1-8、C_1-6、C_1-5、C_1-4Or C_1-3. Examples of the alkyl radical include, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl and sec-butyl. Similarly, the terms alkenyl and alkynyl denote unsaturated straight-chain or branched structures containing, for example, from 2 to 12, preferably from 2 to 6, carbon atoms. In some embodiments, alkenyl or alkynyl is C_2-10、C_2-8、C_2-6、C_2-5、C_2-4Or C_2-3. Examples of alkenyl and alkynyl groups include ethenyl, 2-propenyl, crotyl, 2-isopentenyl, 2- (butadienyl), 2, 4-pentadienyl, 3- (1, 4-pentadienyl), ethynyl, 1-and 3-propynyl, 3-butynyl, and the higher homologs and isomers.

Aralkyl or arylalkyl is meant to denote an alkyl group substituted with an aryl group. An example of arylalkyl is benzyl. Arylalkenyl represents an alkenyl group substituted by an aryl group. Arylalkynyl refers to an alkynyl group substituted with an aryl group. Heteroarylalkyl is meant to denote an alkyl group substituted by a heteroaryl group. Heteroarylalkenyl denotes alkenyl substituted by heteroaryl. Heteroarylalkynyl denotes an alkynyl group substituted by a heteroaryl group. Heterocycloalkyl or heterocycloalkylalkyl is meant to denote an alkyl group substituted with a heterocycle. Cycloalkylalkyl or cyclic alkyl is meant to denote alkyl substituted by cycloalkyl. Examples of cycloalkylalkyl groups include (cyclohexyl) methyl, cyclopropylmethyl, and the like.

The terms alkoxy, alkylamino and alkylthio (or thioalkoxy) are used in their conventional sense to denote those alkyl groups attached to the rest of the molecule via an oxygen atom, an amino group or a sulfur atom, respectively. Thus, terms such as alkoxy and thioalkyl encompass alkyl moieties as defined above attached to an appropriate functionality.

Other suitable substituents which may be used in many carbocyclic rings of the invention, such as cycloaliphatic, aromatic, non-aromatic heterocyclic rings or benzyl groups, include, for example, -OH, halo (-Br, -Cl, -I and-F), -O (aliphatic, substituted aliphatic, benzyl, substituted benzyl, phenyl, substituted phenyl, aromatic or substituted aromatic), -CN, -NO₂、-COOH、-NH₂NH (aliphatic, substituted aliphatic, benzyl, substituted benzyl, phenyl, substituted phenyl, aromatic or substituted aromatic), -N (aliphatic, substituted aliphatic, benzyl, substituted benzyl, phenyl, substituted phenyl, aromatic or substituted aromatic)₂-COO (aliphatic, substituted aliphatic, benzyl, substituted benzyl, phenyl, substituted phenyl, aromatic or substituted aromatic), -CONH₂-CONH (aliphatic, substituted aliphatic, benzyl, substituted benzyl, phenyl, substituted phenyl, aromatic or substituted aromatic)), -SH, -S (aliphatic, substituted aliphatic, benzyl, substituted benzyl, phenyl, substituted phenyl, aromatic or substituted aromatic) and-NH-C ═ NH) -NH₂. The substituted non-aromatic heterocyclic ring, benzylic group, or aromatic group may also have an aliphatic group or a substituted aliphatic group as a substituent. The substituted alkyl or aliphatic groups may also haveThere is a non-aromatic heterocycle, benzyl, substituted benzyl, aromatic group or substituted aromatic group as a substituent. Substituted nonaromatic heterocycles may also have ═ O, ═ S, ═ NH or ═ N (aliphatic, aromatic or substituted aromatic) as substituents. The substituted aliphatic group, substituted aromatic group, substituted non-aromatic heterocycle, or substituted benzyl group may have more than one substituent.

For divalent moieties, such as X and Y, the term "amide bond" means-NHCO-; the term "a thiocarboxamine bond" denotes-NHCS-; the term "sulfonamide" denotes-NHSO₂-; the term "ketone" means-OC-; the term "oxime" denotes — C (═ N-OH) -; the term "urea" denotes-NHCONH-.

"Cyclic alkoxy" means-O- (cycloalkyl). "Heterocycloalkylalkoxy" means-O- (heterocycle). "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group. "Cyclic alkoxyalkyl" refers to an alkyl group substituted with-O- (cycloalkyl). "Heterocycloalkylalkoxyalkyl" refers to an alkyl group substituted with-O- (heterocycle). "alkylthioalkyl" refers to an alkyl group substituted with a thioalkyl group. "Cyclic alkylthioalkyl" refers to alkyl substituted by-S- (cycloalkyl). "Heterocycloalkylthioalkyl" refers to alkyl substituted by-S- (heterocycle). "Mono-or di-substituted amino" denotes-NH wherein one (e.g. mono) or two (e.g. di) hydrogens are replaced by a substituent₂Said substituent being, for example, C_1-8Alkyl, OH, CO- (C)_1-4Alkyl), and the like. "Mono-or di-substituted aminoalkyl" refers to alkyl substituted with mono-or di-substituted amino. "esterified carboxyl" means COOH wherein the hydrogen atom is replaced by a substituent such as C_1-8Alkyl, carbocyclic, heterocyclic, aryl or heteroaryl. "amido" means-CONH₂. "Mono-or di-substituted amide" means-CONH wherein one (e.g., mono) or two (e.g., bis) hydrogens are replaced with a substituent₂Said substituent being, for example, C_1-8Alkyl, OH, CO- (C)_1-4Alkyl), and the like. "Carbamate" means-OCONH₂"Mono-or di-substituted carbamate" means a carbamate wherein one (e.g., mono) or both (e.g., di) hydrogens are replacedsubstituted-OCONH₂Said substituent being, for example, C_1-8Alkyl, OH, CO- (C)_1-4Alkyl), and the like. "sulfonamide" means-SO₂NH₂"Mono-or di-substituted sulfonamides" means-SO wherein one (e.g. mono) or both (e.g. di) hydrogens are replaced by a substituent₂NH₂Said substituent being, for example, C_1-8Alkyl, OH, CO- (C)_1-4Alkyl), and the like. "alkylsulfonyl" means-SO₂- (alkyl). "Cyclic alkylsulfonyl" means-SO₂- (carbocycle). "heterocyclic sulfonyl" means-SO₂- (heterocycles). "arylsulfonyl" means-SO₂- (aryl). "Heteroarylsulphonyl" means-SO₂- (heteroaryl). "alkylcarbonyl" means-CO- (alkyl). "Cyclic alkylcarbonyl" refers to-CO- (cycloalkyl). "Heterocycloalkylcarbonyl" means-CO- (heterocycle). "Arylcarbonyl" means-CO- (aryl). "Heteroarylcarbonyl" means-CO- (heteroaryl). "Thioacylamino" means-CSNH₂. "Arylaminocarbonyl" means-CO-NH- (aryl). "Heteroarylaminocarbonyl" means-CO-NH- (heteroaryl). "Arylamido" means-CO-NH- (aryl). "Heteroarylamido" means-CO-NH- (heteroaryl). "aryl ureido" refers to ureido substituted with aryl. "heteroaryl ureido" refers to ureido substituted with heteroaryl. "aryloxy" means-O- (aryl). "heteroaryloxy" means-O- (heteroaryl). "arylalkoxy" refers to an alkoxy group substituted with an aryl group. "heteroarylalkoxy" means an alkoxy group substituted with a heteroaryl group. "arylamino" means-NH- (aryl). "heteroarylamino" means-NH- (heteroaryl). "hydroxyalkyl" refers to an alkyl group substituted with a hydroxyl group (OH). "aminocarbonylalkyl" refers to an alkyl group substituted with an aminocarbonyl group. "Mono-or di-substituted aminocarbonylalkyl" refers to alkyl substituted with mono-or di-substituted aminocarbonyl. "Alkylcarbonylalkyl" refers to an alkyl group substituted with an alkylcarbonyl group. "Cycloalkylcarbonylalkyl" refers to alkyl substituted by-CO- (cycloalkyl). "Heterocycloalkylcarbonylalkyl" means an alkyl group substituted by-CO- (heterocycle). "Alkylcarboamidoalkyl" refers to an alkyl group substituted with-NH-CO- (alkyl). "cycloalkyl carbonylaminoalkyl"Represents an alkyl group substituted by-NH-CO- (cycloalkyl). "Heterocycloalkylcarboamidoalkyl" refers to an alkyl group substituted with-NH-CO- (heterocycle). "Arylcarbonylaminoalkyl" refers to an alkyl group substituted with-NH-CO- (aryl). "Heteroarylcarbonylaminoalkyl" means an alkyl group substituted by-NH-CO- (heteroaryl). "Arylsulfonylaminoalkyl" means substituted by-NH-SO₂- (aryl) substituted alkyl. "Heteroarylsulphonylaminoalkyl" denotes a radical which is substituted by-NH-SO₂- (heteroaryl) substituted alkyl.

"Spiro" refers to a cycloalkyl group that shares one of its ring-forming atoms with another cycloalkyl or heterocyclyl group. "Spiroheterocycle" means a heterocyclic group that shares one of its ring-forming atoms with another cycloalkyl or heterocyclic group.

The expression "optionally R³And R⁴Can be cyclized to form a bridged bicyclic system having methylene or ethylene groups or heteroatoms selected from N, O and S "meaning R on different atoms³And R⁴Together form a divalent bridging moiety, such as methylene, ethylene, NH, O, S, methylene-O, methylene-S, or methylene-NH.

Unless otherwise indicated, the compounds provided in the above formula are meant to include pharmaceutically acceptable salts, prodrugs, enantiomers, diastereomers, racemic mixtures, crystalline forms, non-crystalline forms, amorphous forms thereof, and solvates thereof.

The term "pharmaceutically acceptable salts" is meant to include salts of the active compounds prepared with relatively nontoxic acids or bases, depending on the particular substituents on the compounds described herein. When the compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral forms of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral forms of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric, hydrobromic, nitric, carbonic, phosphoric, partially neutralized phosphoric, sulfuric, partially neutralized sulfuric, hydroiodic, or phosphorous acids, and the like, as well as salts derived from relatively nontoxic organic acids such as acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids, such as arginine, and salts of organic acids like glucuronic or galacturonic acids. Certain specific compounds of the invention may contain both basic and acidic functionalities, which allow the compounds to be converted into base or acid addition salts. A list of suitable salts is found in Remington's Pharmaceutical sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p.1418 and Journal of Pharmaceutical Science, 66, 2(1977), each of which is incorporated herein by reference in its entirety.

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

As mentioned above, some of the compounds of the present invention possess a chiral or asymmetric carbon atom (optical center) or double bond; racemates, diastereomers, geometric isomers and individual optical isomers are intended to be encompassed within the scope of the present invention.

Some of the compounds of formula I or II can exist in unsolvated as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention. Certain compounds of the present invention may exist in a variety of crystalline or amorphous forms. In general, all physical forms are substantially equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

In addition to salt forms, the present invention also provides compounds that may be in prodrug form. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention. Alternatively, prodrugs can be converted to the compounds of the invention in an ex vivo (exvivo) environment by chemical or biochemical methods. For example, prodrugs can be slowly converted to the compounds of the present invention when placed in a transdermal patch reservoir containing an appropriate enzyme or chemical agent. Functional groups present in the compounds can be modified in such a manner as to produce prodrugs, in such a manner that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compound. Prodrugs include compounds wherein a hydroxy, amino, mercapto or carboxyl group is bonded to any group that, when administered to a mammalian subject, cleaves to form a free hydroxy, amino, mercapto or carboxyl group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol and amine functional groups in the compounds of the present invention. See T.Higuchi and V.Stella, "Pro-drugs as noveldy delivery Systems," Vol.14 of the A.C.S.Symphosis Series and Bioreversible Carriers in Drug delivery, ed.Edward B.Roche, American pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference in their entirety.

The compounds of the invention (including salts, hydrates and solvates thereof) may be prepared using known organic synthesis techniques and may be synthesized according to any of a number of possible synthetic routes.

The reaction to prepare the compounds of the present invention may be carried out in a suitable solvent, which one skilled in the art of organic synthesis can readily select. Suitable solvents may be substantially non-reactive with the starting materials (reactants), intermediates, or products at the temperatures at which the reaction is carried out, e.g., may vary from the freezing temperature of the solvent to the boiling temperature of the solvent. A given reaction may be carried out in one solvent or in a mixture of more than one solvent. Depending on the particular reaction step, a solvent may be selected that is suitable for the particular reaction step.

The preparation of the compounds of the invention may involve the protection and deprotection of various chemical groups. The need for protection and deprotection and the choice of an appropriate protecting group can be readily determined by those skilled in the art. The chemistry of protecting Groups can be found, for example, in T.W.Greene and P.G.M.Wuts, Protective Groups in Organic Synthesis, 3rd.Ed., Wiley & Sons, Inc., New York (1999), which is incorporated herein by reference in its entirety.

The reaction may be monitored according to any suitable method known in the art. For example, the production of the product can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., nuclear magnetic resonance spectroscopy)¹H or¹³C) Infrared spectroscopy, spectrophotometry (e.g. UV-visible) or mass spectrometry, or chromatographic means such as High Performance Liquid Chromatography (HPLC) or thin layer chromatography.

A variety of 4, 4-disubstituted cyclohexanone derivatives can be synthesized using the scheme described in scheme 1. The compounds of formula 1-2 can be prepared by addition of aryl MgX or ArX/BuLi to 1, 4-cyclohexanedione 1-1. Alternatively, compounds of formula 1-2 can be prepared by treating 1, 4-cyclohexanedione mono-ethylene ketal 1-3 with an aryl MgX, ArX/BuLi or heteroaryl H/lithium tetramethylpiperidine, followed by conversion of the ketal in 1-4 to a ketone using an acid, such as aqueous HCl.

Scheme 1

The 4-arylcyclohexanone derivatives of formula 2-3 can be synthesized following the procedure shown in scheme 2. Treatment of intermediates 1-4 with a dehydrating agent such as thionyl chloride/pyridine followed by the use of a catalyst such as Pd-C or PtO₂The resulting olefin is reduced by hydrogenation. Conversion of the ketal in 2-2 by treatment with acid affords the ketone of formula 2-3.

Scheme 2

Alternatively, the compounds of formulae 2-3 can be synthesized according to scheme 3. Reduction of ketone 1-3 using a reducing agent such as sodium borohydride yields alcohol 3-1, which is converted to mesylate 3-2 by treatment with methanesulfonyl chloride. Displacement of the mesylate 3-2 with a heterocycle such as pyrazole, imidazole, triazole or tetrazole affords intermediate 2-2, which is converted to the compound of formula 2-3 by treatment with an acid such as HCl.

Scheme 3

Ar-substituted pyrazoles, imidazoles, triazoles or tetrazoles

Introduction of substituents on the aromatic ring in the ketones of formula 1-2 or 2-3 can be achieved starting from the ketal intermediates 1-4 or 2-2 using the methods described in schemes 4-8. When the aromatic ring in 1-4 or 2-2 carries a cyano group, the ketal 4-1 is hydrolyzed with a base such as sodium or potassium hydroxide to give the carboxylic acid 4-2. Coupling of 4-2 with an amine using a coupling agent, such as BOP, affords amide 4-3. Treatment of 4-3 with an acid such as HCl affords the ketone of formula 4-4.

Scheme 4

When the aromatic ring in the ketal intermediate 1-4 or 2-2 carries a halide, such as bromine or iodine, the halide can be converted to a substituent using the procedures described in scheme 5. Treatment of 5-1 with butyl lithium followed by quenching with an electrophile such as an alkyl halide, aldehyde, ketone, chloroformate, or carbonate affords the R-substituted ketal 5-2. 5-1 andboric acid ArB (OH)₂Suzuki coupling of (Ar ═ aryl or heteroaryl) or coupling of 5-1 to ArZnCl (ArX (X ═ Br, I) can be generated in situ by treatment with butyllithium, followed by treatment with zinc chloride), or in a catalyst such as Ni (CH)₃COCH(OH)CH₃)₂Treatment of 5-1 with iPrMgCl in the presence of-1, 2-bis (diphenylphosphino) ethane followed by coupling with ArX (X ═ Br, I) gave Ar-substituted ketal intermediate 5-4. Treatment of 5-2 and 5-4 with acid affords the corresponding ketones 5-3 and 5-5.

Scheme 5

Alternatively, the ketones of formula 5-5 can be obtained using the scheme depicted in scheme 6. After conversion of the 5-1 to the boronic ester, a palladium catalyst, such as Pd (PPh), is used₃)₄Coupling of the resulting boronic ester 6-1 with ArX (X ═ Br, I) affords Ar-substituted ketal 5-4, which is treated with an acid such as HCl to afford a ketone of formula 5-5.

Scheme 6

When the Ar group in a ketone of formula 1-2 or 2-3 is a 2-thiazole residue, introduction of a substituent at the 5-position in the thiazole can be achieved using the sequence described in scheme 7. Treatment of thiazole 7-1 with butyllithium followed by quenching with 1, 4-cyclohexanedione mono-ethylene ketal 1-3 affords tertiary alcohol 7-2. Treatment of 7-2 with butyl lithium, followed by quenching of the anion 7-3 with an electrophile such as an alkyl halide, aldehyde, ketone, chloroformate, or carbonate, produces the ketal 7-4 with an R substituent at the 5-position on the thiazole. Alternatively, anion 7-3 can be quenched with zinc chloride using a palladium catalyst, such as PdCl₂(PPh₃)₂The resulting intermediate was coupled with ArX (X ═ Br, I) to give the ketal 7-6 with an Ar residue at the 5-position on the thiazole. However, the device is not suitable for use in a kitchenThe ketals 7-4 and 7-6 are then converted to their corresponding ketones of formulae 7-5 and 7-7 by treatment with an acid such as HCl.

Scheme 7

When the Ar group in the ketones of formula 1-2 or 2-3 is a 5-thiazole residue, introduction of the substituent at the 2-position on the thiazole can be achieved using the sequence depicted in scheme 7. Lithiation of 2-trimethylsilyl protected thiazole 8-1 followed by quenching with 1-3 provided intermediate 8-2. After removal of the trimethylsilyl group using TBAF, lithiation of 8-3 followed by quenching with an electrophile such as an alkyl halide, aldehyde, ketone, isocyanate, chloroformate or carbonate affords the 5-R-substituted thiazole derivative 8-4. Treatment of 8-4 with an acid such as HCl affords a ketone of formula 8-5.

Scheme 8

A variety of 3-aminopyrrolidine intermediates can be prepared as shown in schemes 6-17. Coupling of a carboxylic acid of formula 9-1 with a commercially available pyrrolidine derivative of formula 9-2 using a coupling agent, such as BOP, affords amide 9-3. Hydrogenation using an acid, such as TFA or HCl, or using a palladium catalyst, removes the protecting group P (P ═ Boc, benzyl or Cbz) to afford the pyrrolidine intermediates of formula 9-4.

Scheme 9

4-amino-2-methylpyrrolidine derivatives of formula 10-8 can be prepared using the sequence outlined in scheme 10. After Boc protection at amine and TBS protection at hydroxy in trans-4-hydroxy-L-proline methyl ester 10-1, 10-2 was addedThe ester is reduced to an alcohol and the resulting alcohol is converted to a tosylate. Lithium triethylborohydride (LiEt)₃BH) reduction, the demethylation benzene sulfonylation in 10-3 can be realized. The resulting intermediate 10-4 is deprotected using an acid, such as HCl, to remove the Boc and TBS groups. After coupling the resulting amine 10-5 with a carboxylic acid of formula 9-1 using a coupling agent, such as EDC, the hydroxyl group is converted to a mesylate followed by displacement with sodium azide. The resulting azido group is then reduced to the amine by hydrogenation to give the pyrrolidine intermediate of formula 10-8.

Scheme 10

4-Aminopyrrolidine derivatives of formula 11-6 can be prepared according to scheme 11. Alkylation of intermediate 10-2 with alkyl halide (RX) using LHMDS affords R-substituted intermediate 11-1. After reduction of the ester to the alcohol using diisobutylaluminum hydride (DIBAL), the alcohol is converted to tosylate using LiEt₃The resulting tosylate is reduced by BH to give 11-2. Intermediate 11-2 is then converted to the compound of formula 11-6 in a similar manner as described in scheme 10.

Process 111

The 4-aminopyrrolidine derivatives of formula 12-5 can be synthesized using the procedures shown in scheme 12. Intermediate 10-2 is reduced to the alcohol using a reducing agent, such as DIBAL, and the resulting alcohol is alkylated with an alkyl halide (RX) using sodium hydride to give intermediate 12-1. Using procedures similar to those described in scheme 10, the compound of formula 12-5 is obtained from intermediate 12-1.

Scheme 12

4-Aminopyrrolidine derivatives of formula 13-7 can be generated according to scheme 13. Intermediate 10-2 is reduced to an alcohol using a reducing agent, such as DIBAL, and the resulting alcohol is oxidized to an aldehyde using an oxidizing agent, such as Swern oxidation. Addition of the Grignard reagent RMgX to the aldehyde 13-1 is followed by alkylation of the resulting alcohol with an alkyl halide (RX) using sodium hydride. After removal of the Boc and TBS protecting groups in 13-2 or 13-3 using an acid, e.g., HCl, the resulting amine 13-4 is condensed with a carboxylic acid of formula 9-1. Mesylation of the 4-hydroxy group on pyrrolidine followed by displacement of the resulting mesylate with sodium azide and reduction of the azide moiety by hydrogenation affords compounds of formula 13-7.

Scheme 13

4-Aminopyrrolidine derivatives of formula 14-6 can be synthesized using the scheme depicted in scheme 14. After double addition of the Grignard reagent RMgX to the intermediate 10-2, the resulting tertiary alcohol 14-1 is alkylated with an alkyl halide (R' X) to give 14-2. Intermediates 14-1 and 14-2 are then converted to compounds of formula 14-6 in a manner similar to that described in scheme 13.

Scheme 14

The synthesis of 4-aminopyrrolidine derivatives of formula 15-5 is shown in scheme 15. After dehydration of intermediate 14-1 followed by reduction of the olefin by hydrogenation, the resulting intermediate 15-1 is converted to the compound of formula 15-5 in a manner similar to that described in scheme 10.

Scheme 15

The compounds of formula I can be obtained by: the assembly of the aminopyrrolidine derivatives of formula 16-1 with the ketones of formula 16-2 is carried out by reductive amination using a reducing agent such as sodium triacetoxyborohydride, or by hydrogenation followed by treatment of the secondary amine 16-3 obtained via reductive amination with an aldehyde or alkylation with an alkyl halide (RX).

Scheme 16

Alternatively, the compounds of formula I can be prepared using the sequence described in scheme 17. Reductive amination of an aminopyrrolidine derivative of formula 17-1 with a ketone of formula 16-2 affords secondary amine 17-2. After removal of the protecting group P (P ═ Boc, benzyl or Cbz) by hydrogenation using an acid or using a catalyst such as Pd — C, the resulting amine 17-3 is condensed with a carboxylic acid of formula 9-1 to give a compound of formula 17-4.

Scheme 17

Alternatively, the compounds of formula I can be prepared using the sequence depicted in scheme 18. Cyclohexanone 1-2 is reduced with a reducing agent such as lithium aluminum hydride to form cis-diol 18-1. After conversion of the secondary alcohol to a mesylate, the resulting mesylate 18-2 is displaced with an aminopyrrolidine derivative of formula 17-1 to afford a trans 4-amino-1-cyclohexanol derivative of formula 18-3. Removal of the protecting group using an acid or by hydrogenation followed by coupling of the resulting amine with a carboxylic acid of formula 9-1 affords a compound of formula 18-5.

Scheme 18

Alternatively, compounds of formula I can be synthesized according to scheme 19. The mesylate 18-2 is displaced with sodium azide to give the azido intermediate 19-1 which is reduced to the amine by hydrogenation using a catalyst such as Pd-C. Displacement of the mesylate of formula 19-3 with the resulting amine 19-2 or reductive amination of 19-2 with a ketone of formula 19-4 affords compounds of formula 19-5.

Scheme 19

The compounds of the invention are modulators, e.g., antagonists, of the MCP-1 receptor and are capable of inhibiting the binding of MCP-1 to its receptor. Surprisingly, these compounds block T cell migration in vitro and have dramatic effects on inflammatory cell recruitment in a variety of models of inflammatory diseases. Accordingly, the compounds of formula I are useful as therapeutic agents for inflammatory diseases, especially those associated with lymphocyte and/or monocyte accumulation, such as arthritis, rheumatoid arthritis, multiple sclerosis, neuropathic pain, atherosclerosis and graft rejection. In addition, these compounds can be used for the treatment of allergic allergies characterized by basophil activation and eosinophil recruitment, such as asthma and allergic rhinitis, as well as for the treatment of restenosis and chronic or acute immune disorders.

Modulation of chemokine receptor activity as used in the context of the present invention is intended to encompass antagonism, agonism, partial antagonism and/or partial agonism of the activity associated with a particular chemokine receptor, preferably the CCR2 receptor. The term composition as used herein is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts. Pharmaceutically acceptable means that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The compounds of formula I and compositions thereof of the present invention are useful for modulating chemokine receptor activity, particularly CCR 2. Accordingly, the compounds of the present invention are those that inhibit at least one function or characteristic of a mammalian CCR2 protein, such as a human CCR2 protein. The ability of a compound to inhibit this function can be demonstrated in binding assays (e.g., ligand binding or promoter binding), signaling assays (e.g., activation of mammalian G protein, induction of rapid and transient increases in cytosolic free calcium concentration), and/or cellular response functions (e.g., chemotaxis, exocytosis, or stimulation of inflammatory mediator release by leukocytes).

The following examples illustrate the invention and are not intended to be limiting in any way.

Detailed Description

Examples

The reagents and solvents used below are available from commercial sources, such as Aldrich chemical Co. (Milwaukee, Wis., USA). Mass spectrometry results are provided as mass/charge ratios followed by the relative abundance of each ion (in parentheses). In the table, the single M/e values of the M + H (or M-H, as noted) ions containing the most common atomic isotopes are reported. In all cases, the isotopic pattern corresponds to the intended structural formula.

Example 1

Step A

(3-trifluoromethyl-benzoylamino) acetic acid. To a rapidly stirring solution of glycine (15.014g, 0.20mol) in MeCN (400mL) and 2M NaOH (250mL) at 0 deg.C was slowly added 3- (trifluoromethyl) -benzoyl chloride (41.714g, 0.20mol) 75mL Me over 30minCN solution. The cloudy yellow solution was stirred at 0 ℃ for 30 min. The reaction mixture was acidified with 3M HCl to pH 3 followed by MeCN removal on a rotary evaporator. The resulting mixture was then extracted with EtOAc (400mLx 3). The combined organic layers were dried, filtered and concentrated to give a pale yellow solid (48.53g) which was triturated with toluene (500 mL). After filtration, the solid product was washed with cold toluene until the filtrate was colorless. After drying under high vacuum over the weekend, 44.60g (90%) of the product was obtained as a white powder. MS (M + H)⁺)＝248.1.¹H NMR(DMSO-d₆)12.70(br s，1H)，9.17(m，1H)，8.20(dd，2H)，7.94(dd，1H)，7.78(m，1H)，3.97(d，2H).

Step B

[ (3S) -1- ({ [3- (trifluoromethyl) benzoyl)]Amino } acetyl) pyrrolidin-3-yl](iii) carbamic acid tert-butyl ester. To a solution of the carboxylic acid from step A (2.7g, 11mmol) and tert-butyl (3S) -pyrrolidin-3-ylcarbamate (2.0g, 11mmol) in DMF (30mL) cooled in an ice bath was added BOP (5g, 11mmol) followed by triethylamine (3mL, 22 mmol). The mixture was allowed to warm to room temperature and stirred overnight. Ethyl acetate (150mL) was added. The obtained solution is treated with NaHCO₃And brine, three times each, over MgSO₄Drying and concentrating. Chromatography on silica eluting with EtOAc provided 4.4g (96%) of the desired product. MS (M-Boc + H)⁺316.

Step C

N- {2- [ (3S) -3-Aminopyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. The above product (4.2g) was dissolved in 4N HCl/bisAlkane (30 mL). After stirring at room temperature for 1 hour, the solution was concentrated to obtain 4.0g of the title compound. MS (M + H)⁺316.

Step D

8-phenyl-1, 4-dioxaspiro [4.5]]Decan-8-ol. To a solution of 1, 4-cyclohexanone mono-ethylene ketal (8.1g, 50mmol) in THF (20mL) at 10 deg.C was added a 1M solution of phenylmagnesium bromide in THF (70mL, 70 mmol). The resulting mixture was stirred at room temperature for 2 hours, then saturated NH was used₄The Cl solution was quenched. The solution was extracted 3 times with EtOAc. The combined organic phases were washed with brine and MgSO₄Drying and concentrating. Chromatography on silica gel eluting with 40% EtOAc in hexanes provided 9.5g (81%) of the desired product. MS (M + H)⁺234.

Step E

4-hydroxy-4-phenylcyclohexanone. The above product was dissolved in THF (50 mL). Thereto was added 10% HCl/H₂O (50 mL). The solution was stirred at room temperature overnight and extracted three times with EtOAc. The combined extracts were washed with brine and MgSO₄Drying and concentration gave the title compound as a white solid. MS (M + H)⁺191.

Step F

N- (2- { (3S) -3- [ (4-hydroxy-4-phenylcyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. To the pyrrolidine intermediate from step C (0.3g, 0.85mmol) and from step CA solution of the ketone of E (0.16g, 0.85mmol) in THF (5mL) was added Na (OAc)₃BH (0.35g, 2.5mmol) was added followed by triethylamine (0.2mL, 1.5 mmol). The reaction was continued at room temperature overnight and saturated NaHCO was added₃The solution was quenched. The resulting solution was extracted with EtOAc and the EtOAc layer was over MgSO₄Drying and concentrating. Separation on silica gel eluting with 10% to 30% MeOH/EtOAc afforded the cis (fast moving spot) and trans (slow moving spot) isomers of the title compound. MS (M + H)⁺490.0.

Example 2

Step A

8-pyridin-2-yl-1, 4-dioxaspiro [4.5]]Decan-8-ol to a solution of 2-bromopyridine (14g, 88.6mmol) in anhydrous ether (300mL) cooled at-78 deg.C was slowly added a 2.5M solution of n-butyllithium (36 mL). After the addition, stirring was continued at-78 ℃ for 1 hour. To this was slowly added a solution of 1, 4-cyclohexanedione mono-ethylene ketal (15g, 96mmol) in anhydrous ether (300 mL). When the addition was complete, the mixture was allowed to warm to 0 ℃ and stirring was continued for 1 hour. The reaction was quenched by the addition of aqueous ammonium chloride (4.5g) (100 mL). The organic phase was separated and the aqueous phase was extracted 4 times with methylene chloride. The organic phases were combined over MgSO₄Drying and concentrating. Crystallization from EtOAc provided 7g of the desired product. The mother liquor was purified on silica eluting with 10% MeOH/EtOAc to give 3g of the desired product. MS (M + H)⁺236.0.

Step B

4-hydroxy-4- (pyridin-2-yl) cyclohexanone the above product was dissolved in THF (30mL) and 3N HCl in water (30 mL). The mixture was stirred at 50 deg.CFor 3 hours. After cooling to room temperature, NaHCO was added to the solution₃While stirring until no more bubbling. The organic phase was separated and the aqueous layer was extracted three times with EtOAc. The organic phases were combined over MgSO₄Drying and concentrating. The residue was triturated with EtOAc to give 5.5g of the title compound. MS (M + H)⁺192.

Step C

N- (2- { (3S) -3- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The title compound was prepared by reductive amination of the ketone obtained above with the pyrrolidine derivative obtained in step C of example 1 using a procedure analogous to that described in step F of example 1. MS (M + H)⁺491.

Example 3

N- (2- { (3S) -3- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) (methyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. To the N- (2- { (3S) -3- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino group]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide (49mg, 0.1mmol) and formaldehyde (0.3mL, 37% aq.) in THF (2mL) Na (OAc) was added₃BH (64mg, 0.3 mmol). After stirring overnight at room temperature, saturated NaHCO was added₃The solution was quenched. The resulting solution was extracted with EtOAc and the EtOAc layer was dried (MgSO)₄) And (4) concentrating. Purification by preparative HPLC gave the title compound as a TFA salt. MS (M + H)⁺505.

Example 4

Step A

2-bromo-5-bromomethylpyridine 2-bromo-5-methylpyridine (5.00g, 29.1mmol) and N-bromosuccinimide (5.22g, 29.3mmol) were dissolved in carbon tetrachloride (40mL) under nitrogen. Benzoyl peroxide (0.35g, 1.4mmol) was added and the mixture was heated at reflux for 4 hours. The mixture was cooled to room temperature, filtered and concentrated with NaHCO₃/H₂And O washing. The mixture was adsorbed onto silica gel and then chromatographed, eluting with a hexane to 10% ethyl acetate/hexane gradient. Pure fractions were combined and concentrated to give the desired mono-brominated product as a pale yellow solid, 3.60g (49%). LC/MS (M + H)⁺m/z＝249.8，251.8，253.8。

Step B

2-bromo-5- (methoxymethyl) pyridine 2-bromo-5-bromomethyl-pyridine 4(3.58g, 14.3mmol) was dissolved in methanol (20mL) under nitrogen. Sodium methoxide (0.89g, 15.7mmol, 95%) was added, and the mixture was stirred at room temperature. After 3 hours, the methanol was removed by rotary evaporation and the residue was dissolved in dichloromethane and washed with water. The organic extracts were adsorbed onto silica gel and chromatographed. The column was eluted with a gradient of hexane to 20% ethyl acetate/hexane. The pure fractions were combined and concentrated to give the title compound as a colourless oil, 2.62g (90%). LC/MS (M + H)⁺m/z＝202.0.

Step C

4-hydroxy-4- [5- (methoxymethyl) pyridin-2-yl]Cyclohexanone 2-bromo-5- (methoxymethyl) pyridine (2.61g, 12.9mmol) was dissolved in anhydrous THF (40mL) under nitrogen and cooled to-78 deg.C. N-butyllithium (6.20mL, 15.5mmol, 2.5M in hexanes) was added dropwise over 10 minutes to give a black solution. After 15 minutes, 1, 4-dioxa-spiro [4.5] was added dropwise over 2 minutes]A solution of decan-8-one (2.21g, 14.1mmol) in THF was allowed to warm gradually to room temperature over 3 hours. TLC (50% ethyl acetate/hexane) and LC/MS indicated complete conversion. Aqueous HCl (14mL, 6.0M) was added and the mixture was stirred at room temperature for 3h, then NaHCO was used₃/H₂And O neutralizing. The mixture was extracted 3 times with ethyl acetate, the combined extracts were adsorbed onto silica gel and chromatographed. The column was eluted with a gradient of hexane to 40% ethyl acetate/hexane. Pure fractions were combined and concentrated to give the title compound as a pale yellow solid, 1.00g (33%). LC/MS (M + H)⁺m/z＝236.1.

Step D

N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (methoxymethyl) pyridin-2-yl]Cyclohexyl } amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. The title compound was prepared from the ketone of step C using a procedure similar to that described for example 1. MS (M + H)⁺535.

Example 5

Step A

6-bromo-pyridine-3-carbaldehyde 9.48g (40mmol) of 2, 5-dibromopyridine are dissolved in 60mL of THF and 150mL of anhydrous ether. After cooling the solution to-78 deg.C, 16mL of n-butyllithium (2.5M, 40mmol) were slowly added dropwise over 30min via syringe. Stirring at-78 deg.C for 30minAfter this time, N-dimethylformamide (3.5g, 48mmol) was added. The reaction mixture was allowed to warm to room temperature over 2 hours and then quenched by the addition of 10mL of water. The mixture was extracted twice with EtOAc. The combined extracts were dried and concentrated. Flash column, eluting with 30-40% EtOAc in hexane afforded 2.80g of a white solid (28% yield). MS: (M + H)⁺186.0，188.0。

Step B

1- (6-bromopyridin-3-yl) -N, N-dimethylmethylamine. To a solution of titanium tetraisopropoxide (6.4g, 22mmol) and 2.0M dimethylamine in methanol (22mL, 44mmol) was added a solution of 6-bromo-pyridine-3-carbaldehyde (2.10g, 11mmol) in 20mL of methanol. After stirring at r.t. for 5hr, sodium borohydride (0.43g, 11mmol) was added and the mixture was stirred overnight. The reaction was quenched by the addition of 10mL of water and extracted twice with EtOAc. The combined extracts were dried and concentrated. Flash column on, 20-40% methanol in EtOAc and 0.5% NH₄OH eluted to give 1.15g of oil (47% yield). MS: (M + H)⁺214.0，216.0.

Step C

8- {5- [ (dimethylamino) methyl group]Pyridin-2-yl } -1, 4-dioxaspiro [4, 5]]Decan-8-ol. 1- (6-Bromopyridin-3-yl) -N, N-dimethylmethylamine (1.15g, 5.4mmol) was dissolved in 30mL THF and 80mL anhydrous ether. After cooling the solution to-78 deg.C, 2.60mL of n-butyllithium (2.5M, 6.40mmol) were slowly added dropwise over 10min via syringe. After stirring at-78 ℃ for 30 minutes, 1, 4-cyclohexanedione mono-ethylene ketal (1.01g, 6.4mmol) was added. The reaction mixture was allowed to warm to room temperature over 2 hours and then quenched by the addition of 10mL of water. The mixture was extracted twice with EtOAc. The combined extracts were dried and concentrated. On a flash column using 20-40% methanolEtOAc solution and 0.5% NH₄OH eluted to give 0.85g of oil (54% yield). MS: (M + H)⁺293.2.0。

Step D

4- {5- [ (dimethylamino) methyl group]Pyridin-2-yl } -4-hydroxycyclohexanone 8- {5- [ (dimethylamino) methyl]Pyridin-2-yl } -1, 4-dioxaspiro [4, 5]]Decan-8-ol (0.85g, 2.9mmol) was dissolved in 10mL THF and 10mL 2N HCl solution was added. After stirring for 2 hours, saturated NaHCO was added₃The reaction mixture was neutralized to pH 8-9 with aqueous solution and extracted twice with EtOAc. The combined extracts were dried and concentrated to give 0.37g of a white solid (51% yield). MS: (M + H)⁺249.2.

Step E

N- (2- { (3S) -3- [ (4- {5- [ (dimethylamino) methyl group)]Pyridin-2-yl } -4-hydroxycyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The title compound was prepared from the above ketone following the procedure described for example 1. MS (M + H)⁺548。

The following examples 6-13 were prepared in a similar manner to the previous 5 examples.

Example 6

N- [2- ((3S) -3- { [ 4-hydroxy-4- (4-methylphenyl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M+H)⁺504。

Example 7

N- (2- { (3S) -3- [ (4-hydroxy-4-pyridin-3-ylcyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. MS (M + H)⁺491。

Example 8

N- (2- { (3S) -3- [ (4-hydroxy-4-pyridin-4-ylcyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. MS (M + H)⁺491。

Example 9

N- [2- ((3S) -3- { [ 4-hydroxy-4- (5-methylpyridin-2-yl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺505。

Example 10

N-[2-((3S)-3-{[4-hydroxy-4- (4-methylpyridin-2-yl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺505。

Example 11

N- [2- ((3S) -3- { [ 4-hydroxy-4- (6-methylpyridin-2-yl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺505。

Example 12

N- [2- ((3S) -3- { [ 4-hydroxy-4- (6-methoxypyridin-2-yl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺521。

Example 13

N- [2- ((3S) -3- { [ 4-hydroxy-4- (6-methoxypyridin-3-yl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺521。

Example 14

Step A

8- (1, 3-Thiazol-2-Yl) -1, 4-Dioxaspiro [4.5]]Decan-8-ol A solution of N-butyllithium (8.1mL of a 1.6M hexane solution, 12.92mmol) was added to a solution of thiazole (1.0g, 11.75mmol) in THF (10mL) at-78 deg.C while under N₂Stirring the mixture. After stirring at-78 deg.C for 1hr, a solution of 1, 4-cyclohexanedione mono-ethylene ketal (1.84g, 11.75mmol) in THF (10mL) was added via syringe to the lithiated compound solution and stirred at-78 deg.C for 3 hr. Water (5mL) was added, the reaction mixture was warmed to room temperature and extracted with EtOAc (3X). The combined organic layers were dried (MgSO)₄) Filtered, concentrated in vacuo, and chromatographed to give 2.531g of 8- (1, 3-thiazol-2-yl) -1, 4-dioxaspiro [4.5]]Decane-8-ol, yield 89%. MS (EI) (M + H)⁺＝242.2。

Step B

8- (5-methyl-1, 3-thiazol-2-yl) -1, 4-dioxaspiro [4.5]]Decan-8-ol an n-butyllithium solution (5.70mL of a 1.6M hexane solution, 9.12mmol) was added to 8- (1, 3-thiazol-2-yl) -1, 4-dioxaspiro [4.5] at-78 deg.C]Solution of decan-8-ol (1.00g, 4.14mmol) in THF (10mL) while under N₂Stirring the mixture. After stirring at-78 deg.C for 1hr, methyl iodide (0.71mL, 9.12mmol) was added via syringe to the lithiated compound solution at-78 deg.C. The reaction mixture was allowed to warm slowly to room temperature and stirred overnight. Water and EtOAc were added. The aqueous layer was extracted with EtOAc (3 ×). The combined organic layers were washed with saturated NaCl and dried (MgSO)₄) Concentrated and flash chromatographed using 20% EtOAc in hexane to give 0.77g of the title compound in 71% yield. MS (EI) (M + H)⁺＝256.1.

Step C

4-hydroxy-4- (5-methyl-1, 3-thiazol-2-yl) cyclohexanone the mixture of 8- (5-methyl-1, 3-thiazol-2-yl) -1, 4-dioxaspiro [4.5]]A solution of decan-8-ol (1.0g, 4.14mmol) in 20mL THF/3NHCl (1: 1) was stirred at 50 deg.C for 1 hr. After cooling to room temperature, the mixture was taken over Na₂CO₃Work up to pH 8 and extract with EtOAc (3X). The combined organic layers were washed with saturated NaCl solution and dried (MgSO)₄) And concentrated to give 0.82g of 4-hydroxy-4- (5-methyl-1, 3-thiazol-2-yl) cyclohexanone in a yield of 99%. MS (EI) (M + H)⁺＝212.2.

Step D

3- (trifluoromethyl) -N- [2- ((3S) -3- { [ 4-hydroxy-4- (5-methyl-1, 3-thiazol-2-yl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]A benzamide. The title compound was prepared from the ketone of step C using a procedure similar to that described in example 1. Ms (ei): (M + H)⁺511.1.

The following examples 15-16 were prepared in a similar manner to example 14.

Example 15

3- (trifluoromethyl) -N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1-hydroxy-1-methylethyl) -1, 3-thiazol-2-yl]Cyclohexyl } amino) pyrrolidin-1-yl]-2-oxoethyl } benzamide. Ms (ei): (M + H)⁺555.2。

Example 16

3- (trifluoromethyl) -N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (methoxymethyl) -1, 3-thiazol-2-yl)]Cyclohexyl } amino) pyrrolidin-1-yl]-2-oxoethyl } benzamide. Ms (ei): (M + H)⁺541.1。

Example 17

Step A

2- (8-hydroxy-1, 4-dioxaspiro [4.5]]Dec-8-yl) -1, 3-thiazole-4-carboxylic acid an n-butyllithium solution (17.1mL of a 1.6M hexane solution, 27.35mmol) was added to 8- (1, 3-thiazol-2-yl) -1, 4-dioxaspiro [4.5] at-78 deg.C]Decane-8-ol (3.00g, 12.43mmol) in THF (50mL) while under N₂Stirring the mixture. After stirring at-78 deg.C for 1hr, dry ice (10g, 227mmol) was added to the lithiated compound solution, and stirring was carried out at-78 deg.C for 2 hr. Water was added and the solution was allowed to warm to room temperature. The mixture was then treated with 1N HCl to pH 3 to 4 and extracted with EtOAc (3 ×). The combined organic layers were washed with saturated NaCl solution and dried (MgSO)₄) Concentrated and chromatographed (EtOAc to 1% AcOH/EtOAc) to give 3.23g of 2- (8-hydroxy-1, 4-dioxaspiro [4.5]]Decan-8-yl) -1, 3-thiazole-4-carboxylic acid. MS (EI) (M + H)⁺＝286.0.

Step B

2- (8-hydroxy-1, 4-dioxaspiro [4.5]]Decan-8-yl) -N-methyl-1, 3-thiazol-4-ylAn amine. To a stirred mixture of 2- (8-hydroxy-1, 4-dioxaspiro [4.5]]Dec-8-yl) -1, 3-thiazole-4-carboxylic acid (0.30g, 1.05mmol) with methylamine (2M in THF, 2mL, 4mmol) in CH₂Cl₂(10mL) solution Et was added₃N (0.5mL, 3.6mmol) was followed by EDC (0.242g, 1.262mmol) and HOBt (0.193g, 1.26 mmol). The mixture was stirred at room temperature overnight. The reaction mixture was then diluted with EtOAc and saturated Na₂CO₃And a brine wash. The organic layer was dried (MgSO₄) Concentration, flash chromatography (50% EtOAc/hexanes) afforded 0.16g of the title compound in 50% yield. MS (EI) (M + H)⁺＝299.0.

Step C

2- (1-hydroxy-4-oxocyclohexyl) -N-methyl-1, 3-thiazole-4-amide. The title compound was prepared via the conversion of the ketal of step B to a ketone using a procedure similar to that described in step C of example 14. MS (EI) (M + H)⁺＝255.0.

Step D

2- (1-hydroxy-4- { [ (3S) -1- ({ [3- (trifluoromethyl) benzoyl ] amino } acetyl) pyrrolidin-3-yl ] amino } cyclohexyl) -N-methyl-1, 3-thiazole-5-amide. The title compound was prepared from the ketone of step C using the method described for example 1. Ms (ei): (M + H) +553.

The following examples 18-19 were prepared in a similar manner to example 17.

Example 18

N-Ethyl-2- (1-hydroxy-4- { [ (3S) -1- ({ [3- (trifluoromethyl) benzoyl)]Amino } acetyl) pyrrolidin-3-yl]Amino } cyclohexyl) -1, 3-thiazole-5-amide. Ms (ei): (M + H)⁺567.1.

Example 19

N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (pyrrolidin-1-ylcarbonyl) -1, 3-thiazol-2-yl)]Cyclohexyl } -amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. Ms (ei): (M + H)⁺594.1.

Example 20

Step A

8- (1, 3-thiazol-5-yl) -1, 4-dioxaspiro [4, 5]]Decan-8-ol 2-TMS-thiazole (2.5g, 15.89mmol) was added to a solution of N-butyllithium (11.9mL of 1.6M hexane solution, 19.07mmol) in THF (20mL) at-78 deg.C while under N₂Stirring the mixture. After stirring at-78 deg.C for 0.5hr, a solution of 1, 4-cyclohexanedione mono-ethylene ketal (2.48g, 15.89mmol) in THF (20mL) was added via syringe to the lithiated compound solution and stirred at-78 deg.C for 1 hr. Water (5mL) and EtOAc were added and the reaction mixture was warmed to room temperature and extracted with EtOAc (3X). The combined organic layers were dried (MgSO)₄) Filtered and crystallized from EtOAc to give 3.4g of 8- (1, 3-thiazol-5-yl) -1, 4-dioxaspiro [4, 5]]Decane-8-ol, yield 90%. MS (EI) (M + H)⁺＝242.1.

Step B

4-hydroxy-4- [2- (morpholin-4-ylcarbonyl) -1, 3-thiazol-5-yl]Cyclohexanone at-78 deg.C N₂Next, an n-butyllithium solution (2.90mL of a 1.6M hexane solution, 4.64mmol) was added to 8- (1, 3-thiazol-5-yl) -1, 4-dioxaspiro [4, 5]]Decane-8-ol (1.00g, 4.10mmol) in THF (20 mL). After stirring at-78 deg.C for 1hr, 4-morpholinecarbonyl chloride (0.93g, 6.15mmol) was added via syringe to the lithiated compound solution, and stirred at-78 deg.C for 2 hr. Water (5mL) was added and the reaction mixture was allowed to warm to room temperature. The reaction mixture was diluted with water and EtOAc. The aqueous layer was extracted with EtOAc (3 ×). The combined organic layers were washed with brine and dried (Na)₂SO₄) And concentrating to obtain a ketal intermediate. The intermediate was then treated with 20mL THF/1NHCl (1: 1) overnight at room temperature. The reaction solution was washed with Na₂CO₃Adjusted to pH 10 and extracted with EtOAc (3 ×). The combined organic layers were washed with brine and dried (Na)₂SO₄) Concentrated and flash chromatographed using 20% EtOAc in hexane to give 309mg of the title compound. MS (EI) (M + H)⁺＝311.0.

Step C

3- (trifluoromethyl) -N- {2- [ (3S) -3- ({ 4-hydroxy-4- [2- (methoxymethyl) -1, 3-thiazol-5-yl)]Cyclohexyl } amino) pyrrolidin-1-yl]-2-oxoethyl } benzamide. The title compound was prepared from the ketone of step B using a procedure similar to that of example 14. Ms (ei): (M + H)⁺541.1.

The following examples 21-23 were prepared in a similar manner to example 20.

Example 21

3- (trifluoromethyl) -N- [2- ((3S) -3- { [ 4-hydroxy-4- (2-methyl-1, 3-thiazol-5-yl) cyclohexyl]-amino } pyrrolidin-1-yl) -2-oxoethyl]A benzamide. Ms (ei): (M + H)⁺511.1.

Example 22

3- (trifluoromethyl) -N- [2- ((3S) -3- { [4- (2-ethyl-1, 3-thiazol-5-yl) -4-hydroxycyclohexyl)]-amino } pyrrolidin-1-yl) -2-oxoethyl]A benzamide. Ms (ei): (M + H)⁺525.2.

Example 23

N- [2- ((3S) -3- { [ 4-hydroxy-4- (2-isopropyl-1, 3-thiazol-5-yl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. Ms (ei): (M + H)⁺539.2.

Example 24

Step A

8- (5-pyridin-3-yl-1, 3-thiazol-2-yl) -1, 4-dioxaspiro [4.5]]Decan-8-ol n-butyllithium (7.8mL of a 1.6M solution in hexane, 12.45mmol) was added to 8- (1, 3-thiazol-5-yl) -1, 4-dioxaspiro [4, 5-ol at-78 deg.C]Solution of decan-8-ol (1.0g, 4.15mmol) in THF (20mL) while under N₂Stirring the mixture. Stirring at-78 deg.C for 0.5hr, adding 12.5mL of 0.5M ZnCl₂(6.23mmol) in THF. The resulting mixture was stirred at room temperature for 0.5hr, 3-bromopyridine (0.40mL, 4.15mmol) and PdCl were added via syringe₂(PPh₃)₂(0.11g, 0.16mmol) in 5mL THF. After refluxing overnight, 10mL of saturated NH were used₄The reaction was quenched with Cl solution. The aqueous layer was extracted with EtOAc (3 ×). The combined organic layers were dried (MgSO)₄) Filtration, concentration in vacuo, and chromatography gave 0.68g of the title compound in 52% yield. Ms (ei) calculation: (M + H)⁺319.1; actually measuring: 319.1.

step B

N- [2- (3S) - (3- { [ 4-hydroxy-4- (5-pyridin-3-yl-1, 3-thiazol-2-yl) cyclohexyl]Methyl } -pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. The title compound was prepared from the ketal of step A following the procedure described for example 14. Ms (ei): (M + H)⁺574.2.

Example 25

N- [2- ({ (3S) -1- [ 4-hydroxy-4- (5-pyridin-2-yl-1, 3-thiazol-2-yl) cyclohexyl]Pyrrolidin-3-yl } amino) -2-oxoethyl]-3- (trifluoromethyl) benzamide. The title compound was prepared following the procedure described for example 24. Ms (ei): (M + H)⁺574.2.

Example 26

Step A

8-pyridazin-3-yl-1, 4-dioxaspiro [4.5]]Decan-8-ol to a solution of pyridazine (17.7mmol, 1.28mL) in THF (60mL) at-78 deg.C was added 2, 2, 6, 6-tetramethyllithium piperidine (71mmol, 10 g). The reaction was then stirred for 6min and 1, 4-dioxa-spiro [4.5] was added]Decan-8-one (71mmol, 11 g). The reaction was stirred at-78 deg.C for 5hr, at which time the reaction was quenched with ethanol, hydrochloric acid and THF (30mL, 1: 1). The resulting solution was extracted with EtOAc. The organic layers were combined and MgSO₄Drying and concentrating. The residue was purified by flash chromatography to give the desired alcohol (44%, 1.84 g). MS (M + H)⁺237.1.

Step B

4-hydroxy-4-pyridazin-3-ylcyclohexanone to a solution of the product from step A (7.79mmol, 1.84g) in THF (15mL) was added HCl (45mmol, 15 mL). The reaction was stirred overnight, then Na₂CO₃And (6) quenching. The solution was then extracted with EtOAc (3 × 100 mL). The combined organic layers were dried and concentrated in vacuo to afford the desired ketone (780mg, 52%). MS (M + H)⁺193.1.

Step C

N- (2- { (3S) -3- [ (4-hydroxy-4-pyridazin-3-ylcyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The title compound was prepared from the ketone of step B using a procedure similar to that described in example 1. MS (M + H)⁺492.2.

Example 27

N- (2- { (3S) -3- [ (4-hydroxy-4-pyrazin-2-ylcyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The title compound was prepared in a similar manner to example 26. MS (M + H)⁺492.2.

Example 28

Step A

8-pyrimidin-2-yl-1, 4-dioxa-spiro [4.5]]Decan-8-ol (1a) to a solution of 2-bromopyrimidine (0.20g, 1.258mmol) in anhydrous methylene chloride (3.0mL) was added dropwise a 1.6M solution of n-butyllithium in hexane (0.86mL) at-78 ℃. The reaction mixture was stirred at-78 ℃ for 29min and 1, 4-dioxa-spiro [4.5] was added dropwise]Decan-8-one (0.196g, 1.26mmol) in CH₂Cl₂(3mL) of the solution. The reaction was stirred at-78 ℃ for 50min with NH₄Aqueous Cl solution was quenched. After warming to room temperature, the mixture was quenched with CH₂Cl₂Extraction was carried out three times. Combining the extracts over MgSO₄Drying, filtration and concentration in vacuo gave 0.50g of crude product. Purification by silica gel column chromatography eluting with 0 → 50% EtOAc in hexane afforded 0.159g (54%) of the desired product as a light brown-yellow solid. MS (M + H)⁺237.2.

Step B

4-hydroxy-4-pyrimidin-2-ylcyclohexanone to a solution of the product from step A (190mmol, 44g) in THF (200mL) was added HCl solution (300mmol, 100 mL). The reaction was stirred for 2 days, after which the reaction was washed with diethyl ether. The aqueous layer was then quenched with NaOH (50%) to pH 11. The aqueous layer was extracted with EtOAc (6 × 300 mL). The organic layers were combined and MgSO₄Dried and concentrated in vacuo. The reaction was purified via flash chromatography to afford the desired ketone (18g, 49%). MS (M + H)⁺193.1.

Step C

N- (2- { (3S) -3- [ (4-hydroxy-4-pyrimidin-2-ylcyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The title compound was prepared from the ketone of step B using a procedure similar to that of example 1. MS (M + H)⁺492.2.

Example 29

Step A

6-Bromonitonitrile (6-Bromonicotionitrile) 6-chloronicotinonitrile (13.8g, 100mmol) was heated at 145 ℃ in phosphorus tribromide (150mL) for 32 hr. After cooling, the mixture was concentrated in vacuo. Phosphorus tribromide (150mL) was added to the residue and the mixture was heated at 145 ℃ for an additional 32 hr. After cooling, the mixture was concentrated in vacuo and a mixture of ice and water (500mL) was added. Sodium bicarbonate was added to neutralize the mixture and the product was extracted with ethyl acetate (3 × 250 mL). Combining the organic extractsWashed with brine and dried over magnesium sulfate. The solvent was removed in vacuo and the residue was chromatographed (hexane-ethyl acetate) to give 14.9g (81%) of 6-bromonicotinonitrile as a white solid:¹H NMR(400MHz，CDCl₃)7.66(d，J＝11.0Hz，1H)，7.80(dd，J＝3.1，11.0Hz，1H)，8.67(d，J＝3.1Hz，1H)；MS(M+H)⁺m/z＝183.0，185.0.

step B

6- (8-hydroxy-1, 4-dioxaspiro [4.5]]Dec-8-yl) nicotinonitrile 6-bromonicotinonitrile (2g, 11mmol) in 50mL anhydrous THF and 15mL anhydrous hexane under argon in liquid nitrogen-Et₂Cooling to-100 ℃ in an O bath. N-butyllithium (7.5mL, 11mmol, 1.6M hexane solution) was added dropwise so that the internal temperature did not exceed-95 ℃. The orange solution was stirred at-100 ℃ to-95 ℃ for another 10min, then treated dropwise over 10min with a solution of 1, 4-cyclohexanedione mono-ethylene ketal (1.8g, 11mmol) in 55mL of anhydrous THF, while still being careful to maintain the temperature below-95 ℃. The reaction mixture was stirred at-100 ℃ to-95 ℃ for 10min, warmed to 20 ℃ and poured into ice water (400 mL). The organic layer was separated and the aqueous layer was washed with Et₂O (200mL) was extracted twice. The combined organic extracts were then dried over MgSO₄Drying and evaporation gave 2.8g of a white crystalline solid. With Et₂O trituration gives 1.9g (67% yield) of white crystals: MS: (M + H)⁺261.

Step C

6- (8-hydroxy-1, 4-dioxaspiro [4.5]]Decan-8-yl) nicotinic acid 6- (8-hydroxy-1, 4-dioxaspiro [4.5]]Dec-8-yl) nicotinonitrile (1.9g, 7.3mmol) in 50mL of 2-methoxyethanol mixed with 50mL of 2.5N NaOH was heated on a steam bath for 15 hr. The solution was cooled in an ice bath and concentratedHCl is adjusted to pH 7-8 and evaporated to dryness. Water (375mL) was added and the pH adjusted to 2 with HCl. The tan solid was filtered off and washed with water to give 1.92g (6.9mmol, 94% yield) of 6- (8-hydroxy-1, 4-dioxaspiro [4.5]]Decan-8-yl) nicotinic acid: MS: (M + H)⁺280.

Step D

6- (8-hydroxy-1, 4-dioxaspiro [4.5]]Decan-8-yl) -N-methylnicotinamide. At room temperature, the 6- (8-hydroxy-1, 4-dioxaspiro [4.5]]Decan-8-yl) nicotinic acid (560mg, 2mmol), methylamine (1.2mL, 2.0M in THF), BOP reagent (1.07g, 2.4mmol) and 0.8mL (6mmol) triethylamine were dissolved in 15mL DMF. The reaction mixture was stirred at room temperature overnight. Direct chromatography on silica gel (flash chromatography) eluting with 50% ethyl acetate-hexanes provided 410mg (70%) of the desired product, 6- (8-hydroxy-1, 4-dioxaspiro [4.5]]Decan-8-yl) -N-methylnicotinamide: MS: (M + H)⁺293.

Step E

6- (1-hydroxy-4-oxocyclohexyl) -N-methylnicotinamide. At room temperature, the 6- (8-hydroxy-1, 4-dioxaspiro [4.5]]Decan-8-yl) -N-methylnicotinamide (410mg, 1.4mmol) was dissolved in a solvent mixture of 7mL THF and 7mL 1N aqueous HCl. The reaction mixture was then stirred at 60 ℃ for 1 hr. The solution was cooled to room temperature and saturated NaHCO was used₃The aqueous solution is adjusted to pH 7-8. The organic layer was separated and the aqueous layer was extracted twice with EA (20mLx 2). The combined organic extracts were then dried over MgSO₄Drying and evaporation gave an oil residue. Chromatography on silica gel (flash chromatography) eluting with 40% ethyl acetate-hexanes provided 410mg (90%) of the desired product, 6- (1-hydroxy-4-oxocyclohexyl) -N-methylnicotinamide: MS: (M + H)⁺249.

Step F

6- (1-hydroxy-4- { [ (3S) -1- ({ [3- (trifluoromethyl) benzoyl)]Amino } acetyl) pyrrolidin-3-yl]Amino } cyclohexyl) -N-methylnicotinamide. 6- (1-hydroxy-4-oxocyclohexyl) -N-methylnicotinamide (100mg, 0.4mmol) and 126mg (0.4mmol) N- {2- [ (3S) -3-aminopyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide was dissolved in 10.0mL of methylene chloride. To the solution was added 170mg (0.8mmol) of sodium triacetoxyborohydride. The reaction mixture was stirred at room temperature for 2 hr. Direct silica gel chromatography gave 48mg (23%) of the final desired product (top spot on TLC and first peak on HPLC). MS: (M + H)⁺547.

The following examples 30-31 were prepared in a similar manner to example 29.

Example 30

6- (1-hydroxy-4- { [ (3S) -1- ({ [3- (trifluoromethyl) benzoyl)]Amino } acetyl) pyrrolidin-3-yl]Amino } cyclohexyl) -N, N-dimethylnicotinamide. MS (M + H)⁺562.

Example 31

N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (pyrrolidin-1-ylcarbonyl) pyridin-2-yl]Cyclohexyl } -amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl)) A benzamide. MS (M + H)⁺588.

Example 32

Step A

8- (5-Bromopyridin-2-yl) -1, 4-dioxaspiro [4.5]]Decan-8-ol to a solution of 2, 5-dibromopyridine (4.10g, 17mmol) in dry toluene (250mL) was added n-BuLi (1.6M, 12mL) dropwise at-78 ℃. After stirring for 2.5 hours at-78 deg.C, 1, 4-dioxa-spiro [4.5] was added to the reaction mixture]A solution of decan-8-one (2.73g, 17mmol) in methylene chloride (25mL) and the resulting mixture stirred for an additional 1h, slowly warming to room temperature. The reaction mixture was poured into NaHCO₃Aqueous (200mL) then extracted with EtOAc (2 × 50 mL). The organic extracts were combined, washed with brine solution (2 × 50mL) over MgSO₄Dried and concentrated in vacuo. The resulting solid was triturated with ether and the solid collected by filtration. The ether solution was concentrated and the solid was chromatographed on silica gel, eluting with hexane/ethyl acetate (2/1) to give a pale yellow solid. Weight of solids combined: 4.26 g. LCMS: 316.10/314.10(M + H)⁺，100％).¹HNMR：8.6(s，1H)，7.82(d，1H)，7.38(d，1H)，4.6(s，1H)，4.0(m，4H)，2.2(m，4H)，1.7(m，4H).

Step B

4- (5-Bromopyridin-2-yl) -4-hydroxycyclohexanone the title compound was prepared by treating the ketal of step A with aqueous HCl following the procedure described in example 2, step B. MS (M + H)⁺271.

Step C

N- [2- ((3S) -3- [4- (5-bromopyridin-2-yl) -4-hydroxycyclohexyl]Aminopyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. To a 1-necked round bottom flask charged with isopropanol (6mL) was added 4- (5-bromopyridin-2-yl) -4-hydroxycyclohexanone (497.6mg, 1.85mmol), N-2- [ (3S) -3-aminopyrrolidin-1-yl]-2-oxoethyl-3- (trifluoromethyl) -benzamide hydrochloride (651mg, 1.85mol) and triethylamine (0.851mL, 6.11 mol). The resulting mixture was stirred at 25 ℃ for 30 minutes. Sodium triacetoxyborohydride (619mg, 2.78mmol) was then added thereto, and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated and the residue was passed through SiO₂Chromatography, eluting with acetone/methanol (100% to 90%/10%) gave two fractions, F1(404mg) and F2(368mg), a total yield of 73%. LCMS: (M + H)⁺571.1/569.1 (for both isomers). Isomer 1¹H NMR(CD₃OD)8.65(t，1H)，8.21(s，1H)，8.14(d，1H)，8.03(dt，1H)，7.88(d，1H)，7.69(m，2H)，4.23(dd，1H)，4.16(s，1H)，4.10(m，2H)，3.90(m，2H)，3.70(m，2H)，3.60(dd，1H)，3.52(m，2H)，2.55(m，1H)，2.42(m，2H)，2.22(m，3H)，1.80(m，4H).

Example 33

N- {2- [ (3S) -3- ({4- [5- (2-formylphenyl) pyridin-2-yl]-4-hydroxycyclohexyl } -amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. Reacting N- [2- ((3S) -3- [4- (5-bromopyridin-2-yl) -4-hydroxycyclohexyl]Aminopyrrolidin-1-yl) -2-oxoethyl]A solution of (3- (trifluoromethyl) benzamide (30.0mg, 0.0527mmol) and (2-formylphenyl) boronic acid (8.6mg, 0.052mmol) in DMF (0.60mL) and aqueous sodium carbonate (2M, 0.198mL) was usedN₂Degassing for 5 minutes. Then in N₂Adding [1, 1' -bis (diphenylphosphino) ferrocene under washing]-palladium (II) dichloride complex with dichloromethane (1: 1) (2.2mg, 0.0026 mmol). The reaction mixture is treated with N₂Degassed for an additional 5 minutes, and then the tube sealed. The reaction mixture was heated at 130 ℃ for 5 minutes under microwave. After cooling down, the reaction mixture was filtered through a short pad of silica gel and washed with CH₃And (5) CN washing. The resulting solution was acidified with TFA to a pH of 1-2 and then purified by preparative HPLC. The appropriate fractions were lyophilized to give the product (23mg, 53%) as a white powder. MS: (M + H)⁺595.

Example 34

N- (2- (3S) -3- [ (4-hydroxy-4-5- [2- (hydroxymethyl) phenyl)]Pyridin-2-ylcyclohexyl) amino]Pyrrolidin-1-yl-2-oxoethyl) -3- (trifluoromethyl) benzamide bis (trifluoroacetate) N-2- [ (3S) -3- (4- [5- (2-formylphenyl) pyridin-2-yl at 0 deg.C]-4-hydroxycyclohexylamino) pyrrolidin-1-yl]A solution of (3.3mg, 0.004mmol) of (2-oxoethyl-3- (trifluoromethyl) benzamide bis (trifluoroacetate) in methanol (0.50mL) was added sodium borohydride (0.455mg, 0.0120 mmol). The reaction mixture was allowed to warm to room temperature, stirred at room temperature for 60 minutes, then at 60 ℃ for 60 minutes. The mixture was purified by preparative HPLC to give the product as a TFA salt (1.1mg, 33%). LCMS: (M + H)⁺597.2.

Example 35

Step A

8-(4-iodo-phenyl) -1, 4-dioxa-spiro [4.5]Decan-8-ol to a solution of 1, 4-diiodobenzene (16.5g, 50mmol) in THF (350mL) at-78 deg.C over 1h was added n-BuLi (2.5M, 24 mL). After stirring for another 30 minutes, 1, 4-dioxa-spiro [4.5] was added]A solution of decan-8-one (7.8g, 50mmol) in THF (30mL) was stirred for 3 h. TMSCl (5.4g, 50mmol) was added to the mixture and the resulting mixture was allowed to warm to room temperature and stirred at room temperature for 18 hours. The reaction mixture was neutralized to pH 6.0 and extracted with ethyl acetate (3 × 50 mL). The organic extracts were combined, washed with brine solution (2 × 50mL), dried over sodium sulfate and concentrated in vacuo. The residue was chromatographed on silica gel, eluting with hexane/ethyl acetate (95/5-100/0). The appropriate fractions were combined to give 8- (4-iodo-phenyl) -1, 4-dioxa-spiro [4.5]]Decan-8-ol (12g, 66.6%), LCMS: 361.2(M + H)⁺100%), and { [8- (4-iodophenyl) -1, 4-dioxaspiro [4.5]]Decan-8-yl]Oxy } (trimethyl) silane (6g, 27%), LCMS: 433.1(M + H)⁺，100％)。

Step B

8- (4-pyrimidin-2-ylphenyl) -1, 4-dioxaspiro [4.5]]Decan-8-ol to 8- (4-iodo-phenyl) -1, 4-dioxa-spiro [4.5] at room temperature]Decan-8-ol (450.0mg, 1.249mmol) in THF (1.0mL) was added dropwise isopropyl magnesium chloride (2.0M in THF, 1.37mL) and the reaction mixture stirred at room temperature for 30 min. In N₂Next, another flask containing a suspension of nickel acetylacetonate (20mg, 0.06mmol) and 1, 3-bis (diphenylphosphino) propane (26mg, 0.062mmol) in THF (3mL) was charged with 2-bromopyrimidine (199mg, 1.25 mmol). The resulting mixture was stirred at room temperature until clear. The mixture was transferred to the degassed Grignard solution prepared above. The resulting mixture was stirred at room temperature overnight. The reaction mixture was diluted with EtOAc, quenched with water, washed with brine, and washed with Na₂SO₄Drying and concentrating. The residue was columned on silica eluting with hexane/EtOAc (2/1) to affordThe compound (270mg, 69%) was required as a white solid. LCMS: 313.1, (M + H, 100%).¹H NMR(CDCl₃)：8.86(d，2H)，8.46(dd，2H)，7.71(dd，2H)，7.24(t，1H)，4.05(d，4H)，2.30(dt，2H)，2.18(dt，2H)，1.90(m，2H)，1.78(m，2H).

Step C

4-hydroxy-4- (4-pyrimidin-2-ylphenyl) cyclohexanone the title compound was prepared by treating the ketal of step B with aqueous HCl following the procedure described in step B of example 2. MS (M + H)⁺269.

Step D

N- [2- ((3S) -3- [ 4-hydroxy-4- (4-pyrimidin-2-ylphenyl) cyclohexyl]Aminopyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide bis (trifluoroacetate) to a 1-necked round bottom flask charged with methylene chloride (1mL) was added 4-hydroxy-4- (4-pyrimidin-2-ylphenyl) cyclohexanone (50.0mg, 0.186mmol), N-2- [ (3S) -3-aminopyrrolidin-1-yl]-2-oxoethyl-3- (trifluoromethyl) benzamide hydrochloride (65.5mg, 0.186mmol) and triethylamine (85.7 μ L, 0.615 mmol). The resulting mixture was stirred at 25 ℃ for 30 minutes, to which was added sodium triacetoxyborohydride (62.4mg, 0.28mmol) portionwise. The reaction mixture was stirred at room temperature overnight and concentrated. The residue is passed through SiO₂Chromatography eluting with acetone/methanol (100% to 90%/10%) gave two fractions which were further purified by preparative LCMS to give F1(24.2mg) and F2(25.9mg) as white powders in a total yield of 34%, respectively. LCMS of two isomers: 568.2(M + H, 100%).

The following examples 36-37 were prepared in a similar manner.

Example 36

N- [2- ((3S) -3- { [ 4-hydroxy-4- (5-phenylpyridin-2-yl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺567.

Example 37

N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1, 3-thiazol-2-yl) pyridin-2-yl)]Cyclohexyl } amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. MS (M + H)⁺574.

Example 38

Step A

8- (5-pyrimidin-2-ylpyridin-2-yl) -1, 4-dioxaspiro [4.5] decan-8-ol.

A solution of 8- (5-bromopyridin-2-yl) -1, 4-dioxaspiro [4.5] decan-8-ol (168.5g, 0.5363mol) in THF (2000mL) was degassed with nitrogen for 30 min. To the above solution was added dropwise a 2.0M solution of isopropyl magnesium chloride in THF (563mL) over 70min at room temperature. The reaction mixture (light brown) was stirred at 25 ℃ for 180 minutes. Another flask was charged with THF (500mL) and degassed with nitrogen for 10 min. Nickel acetylacetonate (6.9g, 0.027mol) and 1, 2-bis (diphenylphosphino) -ethane (11g, 0.027mol) were added thereto under nitrogen flushing, and after 10 minutes 2-iodopyrimidine (113g, 0.536mol) was added. After stirring for 30 minutes at 25 ℃, the resulting pale green suspension was transferred to the above solution. The reaction mixture was stirred at room temperature overnight and the reaction was found to be complete by HPLC. LC-MS of the desired product: (M + H)314.20 was found. The reaction mixture was used directly for the next reaction.

Step B

4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexanonone.

About half of the THF in the reaction mixture from step a was removed by evaporation under reduced pressure. To the remaining reaction mixture was added 4.00M aqueous HCl (900 mL). After stirring for 1 hour, the mixture was diluted with 1000mL of water and washed with solid Na₂CO₃Neutralizing to pH 8-9. A large amount of yellow solid precipitated out. The solid is filtered off and washed with a solution containing 1% NH₄The aqueous OH solution was washed with ethyl acetate (about 2000mL) until no desired product was detected by TLC. The filtrate was partitioned and the aqueous layer was extracted with ethyl acetate (1200mLx 3). The organic layers were combined, washed with brine, dried over magnesium sulfate and concentrated to half volume. The precipitated solid was filtered and dissolved in dichloromethane (600 mL). The resulting solution was heated to reflux for 30 minutes and filtered. The filtrate was cooled in an ice bath. The precipitated solid was collected by filtration to give 30g of pure product. The mother liquors of the two crystallizations were combined and evaporated. The residue was dissolved in acetonitrile (500 mL). The resulting solution was heated to reflux until all the solids dissolved. Once insoluble material was filtered off, the filtrate was allowed to stand at room temperature and a solid precipitated out. The solid was filtered and suspended in dichloromethane (700 mL). After heating to reflux, the solution was filtered, evaporated to half volume and cooled in an ice bath. The light brown solid that precipitated out was collected by filtration to give a second crop of solid (58 g). MS (M + H)270.2.

Step C

N- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide.

To N- {2- [ (3S) -3-aminopyrrolidin-1-yl]A solution of (E) -2-oxoethyl } -3- (trifluoromethyl) benzamide hydrochloride (22.10g, 47.1mmol) and 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexanone (12.7g, 47.1mmol) in isobutanol (80.0mL) was added triethylamine (19.7mL, 141 mmol). The reaction mixture was cooled in an ice bath and stirred for 30 minutes. To this was added sodium triacetoxyborohydride (11.0g, 51.8mmol) in portions. After stirring at room temperature for 4 hours, the solvent was removed by evaporation under reduced pressure. Adding saturated NaHCO₃Aqueous solution, solution extracted with ethyl acetate (150mLx 3). The combined extracts were washed with brine and dried (Na)₂SO₄) Filtering, and concentrating. The residue was columned on silica gel using ethyl acetate (1% NH)₄Aqueous OH)/methanol (95/5 to 80/20). The appropriate fractions were combined and concentrated to give the title compound as a white powder (17.77 g). MS (M + H)569.

The following examples were prepared in a similar manner.

Example 39

N- (2- { (3S) -3- [ (4- {5- [3- (aminocarbonyl) phenyl)]Pyridin-2-yl } -4-hydroxycyclohexyl) -amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. MS (M + H)⁺610.

Example 40

N- (2- { (3S) -3- [ (4- {5- [2- (aminocarbonyl) phenyl)]Pyridin-2-yl } -4-hydroxycyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. MS (M + H)⁺610.

EXAMPLE 41

N- {2- [ (3S) -3- ({4- [5- (3-acetylphenyl) pyridin-2-yl]-4-hydroxycyclohexyl } amino) -pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. MS (M + H)⁺609.

Example 42

3- [6- (1-hydroxy-4- { [ (3S) -1- ({ [3- (trifluoromethyl) benzoyl)]Amino } acetyl) -pyrrolidin-3-yl]Amino } cyclohexyl) pyridin-3-yl]Benzoic acid, MS (M + H)⁺611.

Example 43

N- (2- { (3S) -3- [ (4-hydroxy-4- {5- [3- (hydroxymethyl) phenyl)]Pyridin-2-yl } cyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide。MS(M+H)⁺597.

Example 44

N- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrimidin-5-ylpyridin-2-yl) cyclohexyl]Amino } -pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺569.

Example 45

N- [2- ((3S) -3- { [4- (3,3' -bipyridin-6-yl) -4-hydroxycyclohexyl ] phenyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺568.

Example 46

N- [2- ((3S) -3- { [4- (3,4' -bipyridin-6-yl) -4-hydroxycyclohexyl ] phenyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺568.

Example 47

N- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrazin-2-ylpyridin-2-yl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺569.

Example 48

N- [2- ((3S) -3- { [ 4-hydroxy-4- (4-iso-isomer)Azol-4-ylphenyl) cyclohexyl]Amino } pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺557.

Example 49

N- {2- [ (3S) -3- ({ 4-hydroxy-4- [4- (1H-imidazol-1-yl) phenyl]Cyclohexyl } amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. MS (M + H)⁺556.

Example 50

4' - (1-hydroxy-4- { [ (3S) -1- ({ [3- (trifluoromethyl) benzoyl) methyl]Amino } acetyl) pyrrolidin-3-yl]Amino } cyclohexyl) biphenyl-2-amide. MS (M + H)⁺609.

Example 51

N- [2- ((3S) -3- { [4- (2' -formylbiphenyl-4-yl) -4-hydroxycyclohexyl]Amino } -pyrrolidin-1-yl) -2-oxoethyl]-3- (trifluoromethyl) benzamide. MS (M + H)⁺594.

Example 52

N- {2- [ (3S) -3- ({ 4-hydroxy-4- [2' - (hydroxymethyl) biphenyl-4-yl]Cyclohexyl } amino) -pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. MS (M + H)⁺596.

Example 53

N- {2- [ (3S) -3- ({4- [5- (3, 5-Dimethyliso-yl)Azol-4-yl) pyridin-2-yl]-4-hydroxycyclohexyl } -amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. MS (M + H)⁺586.

Example 54

N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1, 3-)Azol-2-yl) pyridin-2-yl]Cyclohexyl } amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. MS (M + H)⁺574.

Example 55

Step A

3- (trifluoromethyl) benzaldehyde oxime to a flask containing 3-fluorobenzaldehyde (1.74g, 10mmol) and hydroxylamine hydrochloride (0.76g, 11mmol) in methanol (25mL) was added TEA (0.65g, 11 mmol). The reaction mixture was heated to reflux for 3hr, neutralized to pH 6.0, extracted with ethyl acetate (3 × 20 mL). The organic extracts were combined, washed with brine solution (20mL), dried over sodium sulfate and concentrated in vacuo to give the oxime (1.9g) as a colourless oil. LCMS: (M + H)⁺190.2.

Step B

3- (trifluoromethyl) benzaldoxime to a flask containing a solution of 3- (trifluoromethyl) benzaldoxime (1.89g, 10mmol) in methylene chloride (100mL) was slowly added N-chlorosuccinimide (1.40g, 10.5mmol) at 0 ℃. Heating the reaction mixture to 45 deg.C for 2hr, pouring on ice, and reacting with H₂Diluted O (20mL) and extracted with EtOAc (100 mL). The organic phase is treated with H₂O (2 × 25mL) and brine solution (25mL) were washed, dried over sodium sulfate, and concentrated in vacuo to give oxime (2g, 90%). LCMS: (M + H)⁺224.4.

Step C

3- [3- (trifluoromethyl) phenyl]-4, 5-dihydroisoTo a flask containing a solution of N-hydroxy-3- (trifluoromethyl) benzimidinyl chloride (2.0g, 8.9mmol) and methyl acrylate (0.7g, 8mmol) in methylene chloride (100mL) at 0 deg.C under an inert atmosphere was added TEA (0.90g, 8.8 mmol). The reaction mixture was slowly warmed to ambient temperature, stirred for 20hr, quenched with water (30mL), and extracted with methylene chloride (2 × 50 mL). The organic extracts were combined, washed with brine solution (50mL), dried over sodium sulfate, concentrated in vacuo, and chromatographed on silica gel, eluting with methylene chloride/methanol (100/1-95/5). The appropriate fractions were combined and concentrated in vacuo to give the title compound (2.3g, 100%): LCMS: (M + H)⁺274.2.¹H NMR：(CDCl₃)8.03(s，1H)，7.92(d，1H)，7.71(d，1H)，7.59(dd，1H)，5.28(dd，1H)，3.86(s，3H)，3.71(dd，2H).

Step D

3- [3- (trifluoromethyl) phenyl]-4, 5-dihydroisoOxazole-5-carboxylic acid 3- [3- (trifluoromethyl) phenyl at 0 DEG C]-4, 5-dihydroisoA solution of methyl oxazole-5-carboxylate (2.3g, 8.4mmol) in THF (10mL) was added 2M aqueous sodium hydroxide (10 mL). The reaction mixture was slowly warmed up toAmbient temperature, stir 2hr, neutralize to pH 7 with 2N HCl, extract with ethyl acetate (2 × 50 mL). The organic extracts were combined, washed with brine solution (50mL), dried over sodium sulfate and concentrated in vacuo. The residue was chromatographed on silica, eluting with methylene chloride/methanol (95/5-80/20). The appropriate fractions were combined and concentrated in vacuo to give the title compound (2.18g, 100%) as a white crystalline solid. LCMS: (M-H)^-258.2.

Step E

[ (3S) -1- (3- [3- (trifluoromethyl) phenyl)]-4, 5-dihydroisoAzol-5-ylcarbonyl) pyrrolidin-3-yl]Reaction of 3- [3- (trifluoromethyl) phenyl ] carbamic acid tert-butyl ester at 0 DEG C]-4, 5-dihydroisoA solution of oxazole-5-carboxylic acid (259mg, 1mmol) and tert-butyl (3S) -pyrrolidin-3-ylcarbamate (186mg, 1mmol) in DMF (0.5mL) and methylene chloride (5mL) was added triethylamine (120mg, 1.2mmol) and benzotriazol-1-yloxytris (dimethylamino) -hexafluorophosphate (442mg, 1 mmol). The mixture was allowed to warm to room temperature over 1hr and stirred at room temperature for 1 hr. The mixture was concentrated in vacuo and the residue was chromatographed on silica gel using 1% NH₄Elution of OH in ethyl acetate afforded the desired coupled intermediate (410mg) as a white solid. LCMS: (M + H)⁺428.4.

Step F

(3S) -1- (3- [3- (trifluoromethyl) phenyl)]-4, 5-dihydroisoAzol-5-ylcarbonyl) -pyrrolidin-3-amine hydrochloride to a solution of the intermediate of step E in methylene chloride (5mL) was added 4M HCl in bisAlkane solution (5 mL). After stirring at room temperature for 2hr, the resulting solution was concentrated in vacuo to give the HCl salt of the amine (350mg) as a white solid. LCMS: (M + H)⁺364.4.

Step G

1-pyridin-2-yl-4- [ (3S) -1- (3- [3- (trifluoromethyl) phenyl)]-4, 5-dihydroisoAzol-5-ylcarbonyl) pyrrolidin-3-yl]Aminocyclohexanol to (3S) -1- (3- [3- (trifluoromethyl) phenyl]-4, 5-dihydroisoAzol-5-ylcarbonyl) pyrrolidin-3-amine hydrochloride (178mg, 0.489mmol) and 4-hydroxy-4-pyridin-2-yl-cyclohexanone (95.1mg, 0.498mmol) in methylene chloride (6mL) was added triethylamine (50.3mg, 0.498mmol), NaBH (OAc)₃(120mg, 0.54 mmol). After stirring at room temperature for 2hr, the reaction mixture was neutralized to pH 7 with 1N NaOH and extracted with ethyl acetate (2 × 25 mL). The combined organic extracts were washed with brine solution (20ml), dried over sodium sulfate, concentrated in vacuo, chromatographed on silica gel with 1% NH₄OH in ethyl acetate/methanol (95/5 to 80/20). The appropriate fractions were combined and concentrated in vacuo to afford two fractions of the desired compound: peak 1(100mg) and Peak 2(85 mg). The two fractions were further purified by HPLC on a C18 column using 1% NH₄Elution of OH in water/acetonitrile gave peak 1(68mg) and peak 2(65mg) as a white solid. Two compoundsAll had LCMS: (M + H)⁺503.3. Peak 1 shows two peaks in a 1: 1 ratio in the chiral analytical column. Peak 2 shows two peaks in a 1: 10 ratio in the chiral analytical column.

The following examples 56-58 were prepared in a similar manner to example 55.

Example 56

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({3- [3- (trifluoromethyl) phenyl]-4, 5-dihydroisoAzol-5-yl } carbonyl) pyrrolidin-3-yl]Amino } cyclohexanol. MS (M + H)⁺581.

Example 57

1- {5- [ (dimethylamino) methyl group]Pyridin-2-yl } -4- { [ (3S) -1- ({3- [3- (trifluoromethyl) phenyl]-4, 5-dihydroisoAzol-5-yl } carbonyl) pyrrolidin-3-yl]Amino } cyclohexanol. MS (M + H)⁺560.

Example 58

1-[5-(1，3-Azol-2-yl) pyridin-2-yl]-4- { [ (3S) -1- ({3- [3- (trifluoromethyl) phenyl]-4, 5-dihydroisoAzol-5-yl } carbonyl) pyrrolidin-3-yl]Amino } cyclohexanol MS (M + H)570.3.

Example 59

Step A

(2S, 4R) -N-tert-Butoxycarbonyl-4-hydroxy-2-pyrrolidinecarboxylic acid methyl ester L-trans-4-hydroxyproline methyl ester hydrochloride (25.00g, 138.0mmol) was dissolved in dichloromethane (300mL) and triethylamine (58.0mL, 413 mmol). The solution was cooled to 0 ℃ and di-tert-butyl dicarbonate (33.00g, 151.0mmol) was then added in small portions. After stirring overnight at room temperature, the mixture was concentrated to a viscous white paste. The residue is dissolved in ethyl acetate and the organic layer is successively washed with NH₄Cl/H₂O、NaHCO₃/H₂O and brine wash. The organic extract was passed over MgSO₄Drying, filtration and concentration gave 33.0g (99%) of the desired product as a colorless oil. LC/MS (M + Na)⁺m/z＝267.9.¹H NMR(CDCl₃) 4.50(m, 1H), 4.40(m, 1H), 3.75(s, 3H), 3.43-3.68(m, 2H), 2.30(m, 1H), 1.95-2.15(m, 2H), 1.42 and 1.45(s, 9H).

Step B

2-methyl (2S, 4R) -4- { [ tert-butyl (dimethyl) silyl]Oxy } pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester methyl (2S, 4R) -N-tert-butoxycarbonyl-4-hydroxy-2-pyrrolidinecarboxylate (22.1g, 82.6mmol) was dissolved in anhydrous DMF (100mL) under nitrogen. Imidazole (16.8g, 248mmol) was added and the mixture was cooled to 0 ℃. Tert-butyldimethylsilyl chloride (13.1g, 86.7mmol) was added in small portions and the mixture was allowed to warm to room temperature. After stirring overnight, the mixture was diluted with 300mL ethyl acetate and washed three times with water (500mL, 200mL, 200 mL). The organic extracts were washed a final time with brine and then MgSO₄Drying, filtration and concentration gave 29.5g (99%) of the desired product as a colorless oil. LC/MS (M-Boc + H)⁺m/z＝260.2.¹H NMR(CDCl₃)4.30-4.47(m, 2H), 3.73 and 3.75(s, 3H), 3.60(m, 1H), 3.28-3.45(m, 1H), 2.18(m, 1H), 2.03(m, 1H), 1.42 and 1.47(s, 9H), 0.87(s, 9H), 0.06(s, 6H).

Step C

(2S, 4R) -4- { [ tert-butyl (dimethyl) silyl]Oxy } -2- (hydroxymethyl) -pyrrolidine-1-carboxylic acid tert-butyl ester 2-methyl (2S, 4R) -4- { [ tert-butyl (dimethyl) silyl group under nitrogen]-oxy } pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester (5.00g, 13.91mmol) was dissolved in anhydrous THF (50mL) and cooled to-78 ℃. Diisobutylaluminum hydride solution (31.0mL, 31.0mmol, 1.0M in toluene) was added dropwise over 30 minutes. After stirring for 10min, the mixture was allowed to warm slowly to room temperature, at which time TLC indicated complete conversion. The mixture was diluted with ethyl acetate (200mL) and saturated aqueous sodium potassium tartrate (200 mL). The mixture was stirred vigorously for 30 minutes until two phases were apparent. The aqueous layer was then extracted twice with ethyl acetate and washed with brine. The organic layer was purified over MgSO₄Drying, filtration and concentration gave 4.91g of crude alcohol as a pale yellow oil. LC/MS (M-Boc + H)⁺m/z＝232.2.¹H NMR(CDCl₃).4.88(d，1H)，4.27(bs，1H)，4.14(m，1H)，3.69(t，1H)，3.54(m，1H)，3.42(d，1H)，3.34(dd，1H)，1.96(m，1H)，1.58(m，1H)，1.47(s，9H)，0.87(s，9H)，0.06(s，6H).

Step D

(2S, 4R) -4- { [ tert-butyl (dimethyl) silyl]Oxy } -2- ({ [ (4-methylphenyl) -sulfonyl)]-oxy } methyl) pyrrolidine-1-carboxylic acid tert-butyl ester (2S, 4R) -4- { [ tert-butyl (dimethyl) silyl]Tert-butyl oxy } -2- (hydroxymethyl) pyrrolidine-1-carboxylate (4.91g, 14.8mmol) was dissolved in dichloromethane (70 mL). Triethylamine (5.8mL, 41.7mmol) followed by p-toluenesulfonyl chloride (3.18g, 16.7mmol) was added and the mixture was stirred at room temperature overnight. TLC revealed about half conversion. Pyridine (3.4mL, 41mmol) was added to the mixture, which turned dark orange after 20 minutes. After a further two days, the mixture was diluted with ethyl acetate and the organic layer was successively diluted with NaHCO₃/H₂O、NH₄Cl/H₂O, water and brine. The organic extract was passed over MgSO₄Dried, filtered, concentrated to a red oil and chromatographed on silica gel (10% to 20% ethyl acetate/hexane). The pure fractions were combined to give the tosylate as a yellow oil, 6.32g (93%, 2 steps).¹H NMR(CDCl₃)7.77(d, 2H), 7.34(t, 2H), 4.30(m, 2H), 4.10(m, 2H), 3.30(m, 2H), 2.45(s, 3H), 1.97(m, 2H), 1.41 and 1.37(s, 9H), 0.85(s, 9H), 0.06(s, 6H).

Step E

(2R, 4R) -4- { [ tert-butyl (dimethyl) silyl group]Oxy } -2-methylpyrrolidine-1-carboxylic acid tert-butyl ester under nitrogen, (2S, 4R) -4- { [ tert-butyl (dimethyl) silyl]Oxy } -2- ({ [ (4-methylphenyl) -sulfonyl)]-oxy } methyl) pyrrolidine-1-carboxylic acid tert-butyl ester (6.32g, 1301mmol) was dissolved in THF (50mL) and cooled to 0 deg.C. Lithium triethylborohydride solution (Super Hydride, 14.3mL, 1.0M THF solution) was added dropwise, and the mixture was allowed to warm slowly to room temperature. After 2 hours, TLC revealed half of the conversion. More lithium triethylborohydride solution (12.0mL) was added and the solution was stirred at room temperature overnight. With NaHCO₃/H₂Diluted with O and extracted twice with ethyl acetate. By NH₄Cl/H₂The organic layer was washed with brine. Over MgSO₄The organic extract was dried, filtered and concentrated to give a colorless oil. Chromatography on silica gel eluting with 10% ethyl acetate/hexane. The pure fractions were combined to give the desired product as a colorless oil, 3.74g (91%). LC/MS (M + Na)⁺m/z＝338.2.¹H NMR(CDCl₃)4.34(m，1H)，3.95(m，1H)，3.35(m，2H)，1.98(m，1H)，1.65(m，1H)，1.47(s，9H)，1.20(bs，3H)，0.87(s，9H)，0.06(s，6H).

Step F

(3R, 5R) -5-Methylpyrrolidin-3-ol hydrochloride the (2R, 4R) -4- { [ tert-butyl (dimethyl) silyl group under nitrogen]Tert-butyl oxy } -2-methylpyrrolidine-1-carboxylate (3.74g, 11.85mmol) was dissolved in anhydrous THF (20 mL). Hydrogen chloride solution (40mL, 4.0M 1, 4-bis) was addedAlkane solution), the mixture was stirred at room temperature for 4 hours. The solution was concentrated on a rotary evaporator to an oil, azeotroped with toluene and pumped under vacuum to give the hydrochloride salt as an off-white solid, 1.80g (100%) and used in the next step without further purification.¹H NMR(CD₃OD)4.54(m，1H)，3.95(m，1H)，3.44(dd，1H)，3.18(d，1H)，2.19(dd，1H)，1.76(m，1H)，1.44(d，3H).

Step G

N- {2- [ (2R, 4R) -4-hydroxy-2-methylpyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoro-methyl) benzamide. (3R, 5R) -5-methylpyrrolidine-3-ol hydrochloride (1.80g, 13mmol) was dissolved in dichloromethane (50mL) and diisopropylethylamine (2.1mL, 12.0mmol) under nitrogen. (3-trifluoromethyl-benzoylamino) -acetic acid (2.93g, 11.85mmol) was added followed by EDC (3.41g, 17.8mmol) and the mixture was stirred at room temperature for 4 h. The mixture is treated with NH₄Cl/H₂Diluted with O and extracted twice with ethyl acetate. Combining the extracts, adding NaHCO₃/H₂O and brine, over MgSO₄Dried, filtered and concentrated to give a dark orange oil. Chromatography on silica gel eluting with ethyl acetate to 5% methanol/ethyl acetate afforded the coupled product as a light orange solid, 3.19g (81%, 2 steps). LC/MS (M + H)⁺m/z＝331.1.¹H NMR(CDCl₃Major rotamers) 8.12(s, 1H), 8.01(d, 1H), 7.76(d, 1H), 7.57(t, 1H), 7.50(m, 1H), 4.56(m, 1H), 4.34(m, 1H), 4.23(m, 1H), 4.11(m, 1H), 3.61(dd, 1H), 3.51(d, 1H), 2.71(d, 1H), 2.17(m, 1H), 1.81(m, 1H), 1.32(d, 3H).

Step H

(3R, 5R) -5-methyl-1- ({ [3- (trifluoromethyl) benzoyl) methyl]Amino } acetyl) pyrrolidin-3-yl methanesulfonate to N- {2- [ (2R, 4R) -4-hydroxy-2-methylpyrrolidin-1-yl at 0 ℃ under nitrogen]A solution of (E) -2-oxoethyl } -3- (trifluoromethyl) benzamide (1.50g, 4.54mmol) in dichloromethane (30mL) and pyridine (1.83mL, 22.7mmol) was added methanesulfonyl chloride (0.42mL, 5.45mmol) dropwise. After stirring at 0 ℃ for 2 hours, the reaction was allowed to slowly warm to room temperature and stirred overnight. The mixture is washed with NaHCO₃/H₂Diluted with O and extracted with ethyl acetate. An organic layer is formedBy NH₄Cl/H₂O and brine, over MgSO₄Drying, filtration and concentration gave the mesylate as a brown oil, 1.87g (100%). LC/MS (M + H)⁺m/z＝409.0.¹H NMR(CDCl₃Major rotamers) 8.12(s, 1H), 8.01(d, 1H), 7.78(d, 1H), 7.59(t, 1H), 7.29(bs, 1H), 5.33(m, 1H), 4.37(m, 1H), 4.18(m, 2H), 3.86(d, 1H), 3.76(dd, 1H), 3.08(s, 3H), 2.51(m, 1H), 1.94(m, 1H), 1.38(d, 3H).

Step I

N- {2- [ (2R,4S) -4-azido-2-methylpyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. To a solution of the crude mesylate (1.87g) in anhydrous DMF (20mL) was added sodium azide (1.50g, 22.7 mmol). The mixture was stirred at 60-65 ℃ for 5 hours and then at 50 ℃ for 20 hours. Ethyl acetate was added. The organic layer was separated, washed twice with water, then brine, and MgSO₄Dried, filtered and concentrated to an orange oil. Chromatography on silica gel eluting with 80% ethyl acetate/hexanes afforded the azide as a yellow oil, 1.33g (82%). LC/MS (M + H)⁺m/z＝356.1.¹H NMR(CDCl₃Major rotamers) 8.12(s, 1H), 8.00(t, 1H), 7.77(d, 1H), 7.58(t, 1H), 7.37(bs, 1H), 4.35(m, 2H), 4.17(m, 2H), 3.73(dd, 1H), 3.50(d, 1H), 2.39(m, 1H), 1.87(d, 1H), 1.43(d, 3H).

Step J

N- {2- [ (2R,4S) -4-amino-2-methylpyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. Reacting N- {2- [ (2R,4S) -4-azido-2-methylpyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide (1.33g, 3.74mmol) was dissolved in ethanol (50mL), and then 10% Pd-C (130mg) was added to the solution. The flask was purged with hydrogen and then stirred using a balloon under hydrogen atmosphere for 4 hours, at which time TLC indicated complete consumption of starting material. The reaction was then flushed with nitrogen, filtered through celite on a glass frit (glass frit) and washed with methanol. The filtrate was concentrated to give the desired amine as a dark brown oil, 1.21g (98%). LC/MS (M + H)⁺m/z＝330.1.¹H NMR(CDCl₃)8.12(s，1H)，8.02(d，1H)，7.77(d，1H)，7.58(t，1H)，7.37(bs，1H)，4.16(m，3H)，3.72(m，1H)，3.61(m，1H)，3.15(m，1H)，2.44(m，1H)，1.70-1.20(m，3H)，1.43(d，3H)；¹⁹F NMR(CDCl₃)-63.12(s).

Step K

N- (2- { (2R,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino]-2-methyl-pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. Mixing N- {2- [ (2R,4S) -4-amino-2-methyl pyrrolidine-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide (200mg, 0.607mmol) and 4-hydroxy-4-pyridin-2-yl-cyclohexanone (116mg, 0.607mmol) were dissolved in 2-propanol (10 mL). After stirring for 30min, sodium triacetoxyborohydride (257mg, 1.21mmol) was added and the mixture was stirred at room temperature overnight. TLC indicated complete conversion to the desired product, with a ratio of the two isomers of about 1: 1. The reaction mixture was chromatographed on silica gel eluting with dichloromethane to 10% methanol/dichloromethane/0.5% ammonium hydroxide to give 229mg (75%) as a mixture of isomers.¹H NMR(CDCl₃Mixture of isomers) 8.53(M, 1H), 8.13(bs, 1H), 8.02(d, 1H), 7.75(M, 2H), 7.58(t, 1H), 7.40(M, 2H), 7.22(M, 1H), 4.05-4.38(M, 3H), 3.80(M, 1H), 3.56(M, 1H), 3.42(M, 1H), 3.19(M, 1H), 3.04(M, 1H), 2.65(M, 1H), 2.47(M, 1H), 2.16(M, 2H), 1.40-2.00(M, 7H), 1.43(d, 3H), LCMS (M + H)⁺: the higher Rf isomer m/z 505.2; the lower Rf isomer m/z is 505.2.

Example 60

Step A

(2S, 4R) -4- { [ tert-butyl (dimethyl) silyl]Oxy } -2- (methoxymethyl) -pyrrolidine-1-carboxylic acid tert-butyl ester methyl iodide (0.85mL, 13.6mmol) was added to (2S, 4R) -4- { [ tert-butyl (dimethyl) silyl]Tert-butyl oxy } -2- (hydroxymethyl) pyrrolidine-1-carboxylate (1.50g, 4.52mmol) in anhydrous DMF (15 mL). Sodium hydride (0.22g, 5.42mmol, 60% dispersion in mineral oil) was added in portions and the mixture was stirred at room temperature overnight. The mixture was diluted with ethyl acetate. The organic layer was separated, washed twice with water, then brine, and MgSO₄Drying, filtration and concentration gave 1.51g (96%) of methyl ether as a yellow oil. LC/MS (M-Boc + H)⁺m/z＝246.2.¹H NMR(CDCl₃)4.38(m，1H)，4.05(m，1H)，3.50(m，2H)，3.25-3.45(m，2H)，3.34(s，3H)，1.87-2.06(m，2H)，1.47(s，9H)，0.87(s，9H)，0.06(s，6H).

Step B

N- {2- [ (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino]-2- (methoxymethyl) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. The title compound was prepared from the intermediate of step a following the procedure described for example 59. Higher Rf isomer: LCMSm/z 535.2(M + H);¹H NMR(CDCl₃)8.53(d，1H)，8.12(s，1H)，8.03(d，1H)，7.77(m，1H)，7.72(m，1H)，7.58(t，1H)，7.47(m，1H)，7.34(m，1H)，7.21(m，1H)，4.90(m，1H)，4.12-4.47(m，4H)，3.89(dd，1H)，3.79(dd，1H)，3.54(m，2H)，3.38(s，3H)，3.03(m，1H)，2.40(m，1H)，2.18(m，3H)，1.90(m，1H)，1.75(m，1H)，1.60(m，2H)，1.50(m，2H)；¹⁹F NMR(CDCl₃) -63.11(s) lower Rf isomer: LCMS (M + H)⁺m/z＝535.2；¹H NMR(CDCl₃)8.53(d，1H)，8.12(s，1H)，8.02(d，1H)，7.78(m，1H)，7.72(m，1H)，7.58(t，1H)，7.42(m，1H)，7.34(m，1H)，7.21(m，1H)，4.12-4.48(m，4H)，3.83(m，2H)，3.68(m，1H)，3.56(m，1H)，3.38(s，3H)，2.72(m，1H)，2.38(m，1H)，1.60-2.20(m，10H)；¹⁹F NMR(CDCl₃)-63.12(s).

Example 61

N- (2- { (2S,4S) -2- (ethoxymethyl) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) -amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The title compound was prepared following the procedure described for example 60. Higher Rf isomer: LCMS (M + H)⁺m/z＝549.1；¹H NMR(CDCl₃)8.51(m, 1H), 8.10(m, 1H), 7.99(m, 1H), 7.70(m, 2H), 7.32-7.60(m, 3H), 7.18(m, 1H), 4.03-4.47(m, 3H), 3.22-3.91(m, 5H), 3.04(m, 1H), 1.70-2.47(m, 7H), 1.51(m, 4H), 1.21(m, 4H). lower Rf isomer: LCMS (M + H)⁺m/z＝549.1；¹H NMR(CDCl₃)8.52(m，1H)，8.11(m，1H)，8.00(m，1H)，7.73(m，2H)，7.55(m，1H)，7.39(m，2H)，7.20(m，1H)，4.11-4.48(m，3H)，3.46-3.88(m，5H)，3.21(m，1H)，2.63(m，1H)，2.38(m，1H)，1.55-1.98(m，10H)，1.20(m，3H).

Example 62

Step A

(2S, 4R) -4- { [ tert-butyl (dimethyl) silyl]Oxy } -2- (1-hydroxy-1-methylethyl) pyrrolidine-1-carboxylic acid tert-butyl ester at 0 deg.C to 2-methyl (2S, 4R) -4- { [ tert-butyl (dimethyl) silyl]A solution of 1-tert-butyl oxy } pyrrolidine-1, 2-dicarboxylate (1.00g, 2.78mmol) in anhydrous THF (20mL) was added dropwise over a period of 5 minutes to a solution of methylmagnesium bromide (2.0mL, 6.0mmol, 3.0M in ether). After stirring for 4 hours, the mixture is allowed to warm to room temperature and is quenched with NH₄Cl/H₂O was quenched and extracted twice with ethyl acetate. The organic extract was passed over MgSO₄Drying, filtration and concentration gave 1.00g (100%) of the title compound as a white solid.¹H NMR(CDCl₃)5.85(s，1H)，4.25(s，1H)，4.08(t，1H)，3.67(d，1H)，3.18(d，1H)，1.94(m，1H)，1.60(m，1H)，1.45(s，9H)，1.15(s，3H)，1.05(s，3H)，0.87(s，9H)，0.06(s，6H).

Step B

N- (2- { (2S,4S) -2- (1-hydroxy-1-methylethyl) -4- [ (trans-4-hydroxy-4-pyridin-2-ylcyclohexyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The title compound was prepared from the alcohol of step A following the procedure described for example 59. Higher Rf isomer: LCMS (M + H)⁺m/z＝549.3；¹H NMR(CDCl₃)8.53(m，1H)，8.13(s，1H)，8.01(d，1H)，7.78(d，1H)，7.74(t，1H)，7.59(t，1H)，7.48(d，1H)，7.32(m，1H)，7.22(m，1H)，4.19-4.40(m，3H)，3.98(dd，1H)，3.49(m，2H)，3.29(m，1H)，3.08(m，1H)，2.10-2.45(m，8H)，1.71(m，2H)，1.24(s，3H)，1.21(s，3H)；¹⁹F NMR(CDCl₃)-63.12(s) lower Rf isomer: LCMS (M + H)⁺m/z＝549.3；¹H NMR(CDCl₃)8.52(d，1H)，8.12(s，1H)，8.01(d，1H)，7.77(d，1H)，7.73(m，1H)，7.59(t，1H)，7.40(d，1H)，7.37(m，1H)，7.22(m，1H)，5.14(bs，1H)，4.39(m，1H)，4.33(m，1H)，4.20(m，1H)，3.97(m，1H)，3.72(m，1H)，3.40(m，1H)，2.74(m，1H)，1.70-2.35(m，12H)，1.24(s，3H)，1.21(s，3H)；¹⁹F NMR(CDCl₃)-63.12(s).

Example 63

N- (2- { (2S,4S) -2- [ 1-hydroxyethyl]-4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) -amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The title compound was prepared in a similar manner to example 62. MS (M + H)⁺535.

Example 64

N- {2- [ (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino]-2- (1-methoxy-1-methylethyl) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. In accordance with example 60The process is carried out from (2S, 4R) -4- { [ tert-butyl (dimethyl) silyl]The title compound was prepared starting from tert-butyl oxy } -2- (1-hydroxy-1-methylethyl) pyrrolidine-1-carboxylate. Higher Rf isomer: LC/MS (M + H)⁺m/z＝563.3；¹H NMR(CDCl₃)8.55(m, 1H), 8.14(m, 1H), 8.04(m, 1H), 7.74(m, 2H), 7.38-7.63(m, 3H), 7.22(m, 1H), 5.42-5.80(bs, 1H), 4.84(bs, 1H), 4.15-4.43(m, 3H), 3.96(m, 1H), 3.42(m, 1H), 3.22(m, 4H), 3.02(m, 1H), 1.89-2.34(m, 6H), 1.46-1.67(m, 4H), 1.22(m, 6H). lower Rf isomer: LC/MS (M + H)⁺m/z＝563.3；¹H NMR(CDCl₃)8.53(m，1H)，8.15(m，1H)，8.03(m，1H)，7.74(m，2H)，7.35-7.61(m，3H)，7.22(m，1H)，3.87-4.43(m，4H)，3.50(m，1H)，3.21(m，4H)，2.64(m，1H)，2.27(m，1H)，1.67-1.98(m，9H)，1.22(m，6H).

Example 65

N- (2- { (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino]-2- [ (1S) -1-methoxyethyl group]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The title compound was prepared in a similar manner to example 64. MS (M + H)⁺549.

Example 66

Step A

2-methyl (4R) -4- { [ tert-butyl esterRadical (dimethyl) silyl]Oxy } -2-methylpyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester to 2-methyl (2S, 4R) -4- { [ tert-butyl (dimethyl) silyl ester at-78 deg.C]Oxy } pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester (5.11g, 14.2mmol) in anhydrous THF (60mL) was added dropwise lithium bis-trimethylsilylamide (17.0mL, 17.0mmol, 1.0M in THF). After stirring for 30min, methyl iodide (1.77mL, 28.4mmol) was added. The mixture was stirred at-78 ℃ for 1 hour, warmed to 0 ℃ for 1 hour, and finally with NaHCO₃/H₂And quenching O. The resulting mixture was extracted twice with ethyl acetate. Combining the extracts over MgSO₄Drying, filtering and concentrating. The residue was chromatographed on silica gel eluting with hexane to 5% ethyl acetate/hexane to give 2.66g (50%) of a mixture of product isomers as a colorless oil. LC/MS (M-Boc + H)⁺m/z＝274.1.¹H NMR(CDCl₃)4.38(m，1H)，3.71(m，4H)，3.36(m，1H)，1.84-2.35(m，2H)，1.61(m，3H)，1.44(m，9H)，0.88(m，9H)，0.07(m，6H).

Step B

N- (2- { (4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino]-2, 2-dimethyl-pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. Following the procedure described in example 59, starting from 2-methyl (4R) -4- { [ tert-butyl (dimethyl) silyl]Oxy } -2-methylpyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester the title compound was prepared. LC/MS (M + H)⁺m/z＝519.2；¹H NMR(CD₃OD, bis-trifluoroacetate salt) 8.51(m, 1H), 8.18(m, 2H), 7.63-7.90(m, 4H), 7.27(m, 1H), 4.15(dd, 2H), 3.98(m, 1H), 3.55(m, 1H), 3.28(m, 2H), 2.92(m, 1H), 2.38(m, 2H), 1.96-2.20(m, 3H), 1.50-1.79(m, 7H), 1.42(s, 3H) lower Rf isomer: LC/MS (M + H)⁺m/z＝519.2；¹H NMR(CD₃OD, bis-trifluoroacetate salt)8.49(m，1H)，8.21(m，1H)，8.14(m，1H)，7.65-7.90(m，4H)，7.25(m，1H)，4.10(m，3H)，3.72(m，1H)，3.28(m，2H)，2.73(m，1H)，2.10(m，3H)，1.82(m，2H)，1.73(m，4H)，1.58(s，3H)，1.45(s，3H).

Example 67

Step A

L-trans-4-hydroxyproline methyl ester hydrochloride (9.70g, 54.0mmol) was dissolved in anhydrous THF (180mL) and triethylamine (7.53mL, 54.0 mmol). To this solution was slowly added a solution of N- (benzyloxycarbonoxy) succinimide (13.5g, 54.0mmol) in THF (70 mL). After stirring at room temperature overnight, the mixture was diluted with ethyl acetate and the organic layer was washed successively with water and brine. Subjecting the organic extract to Na₂SO₄Drying, filtering and concentrating. The residue was chromatographed on silica gel (30% to 70% ethyl acetate/hexane) to give 12.8g (85%) of the desired product as a colorless oil. LC/MS (M + H)⁺m/z＝280.0；¹H NMR(CDCl₃)7.33(m, 5H), 5.00-5.25(m, 2H), 4.52(m, 2H), 3.69(m, 2H), 3.56 and 3.78(s, 3H), 2.05-2.40(m, 2H).

Step B

2-methyl (2S, 4R) -4- (benzyloxy) pyrrolidine-1, 2-dicarboxylic acid 1-benzyl ester 2-methyl (2S, 4R) -4-hydroxypyrrolidine-1, 2-dicarboxylic acid 1-benzyl ester (6.60g, 23.6mmol) was dissolved in anhydrous THF (100mL) and cooled to 0 deg.C under nitrogen. Hydrogenation is added portionwiseSodium (1.04g, 26.0mmol, 60% dispersion in mineral oil) and the mixture was stirred for 15 minutes. Tetra-n-butylammonium iodide (0.40g, 1.0mmol) and benzyl bromide (3.15mL, 26.0mmol) were added and the mixture was stirred at 0 ℃ for 1 hour and then at room temperature for 1 hour. The mixture was diluted with ethyl acetate. The organic layer was washed with water, then brine, and MgSO₄Drying, filtering and concentrating. The residue was chromatographed on silica gel (20% to 50% ethyl acetate/hexane) to give 4.21g (48%) of benzyl ether. LC/MS (M + H)⁺m/z＝370.2；¹H NMR(CDCl₃) 7.34(m, 10H), 5.13(m, 2H), 4.51(m, 3H), 4.20(m, 1H), 3.68(m, 2H), 3.54 and 3.78(s, 3H), 2.45(m, 1H), 2.11(m, 1H).

Step C

Benzyl (2S, 4R) -4- (benzyloxy) -2- (1-hydroxy-1-methylethyl) pyrrolidine-1-carboxylate 1-benzyl 2-methyl (2S, 4R) -4- (benzyloxy) pyrrolidine-1, 2-dicarboxylate (4.21g, 11.4mmol) was dissolved in anhydrous THF (20mL) under nitrogen and cooled to 0 ℃. Methylmagnesium bromide solution (8.4mL, 25mmol, 3.0M in ether) was added dropwise. After stirring at 0 ℃ for 12 hours, the mixture is warmed to room temperature and quenched with NH₄Cl/H₂O was quenched and extracted twice with ethyl acetate. The organic extracts were washed with brine, over Na₂SO₄Drying, filtering and concentrating. The residue was chromatographed on silica gel (20% to 30% ethyl acetate/hexane) to give 2.47g (59%) of the alcohol as a viscous oil. LC/MS (M + H)⁺m/z＝370.1；¹H NMR(CDCl₃)7.33(m，10H)，5.55(bs，1H)，5.20(s，2H)，4.50(s，2H)，4.19(m，1H)，4.05(m，2H)，3.31(m，1H)，2.27(m，1H)，1.73(m，1H)，1.21(s，3H)，1.13(s，3H).

Step D

Benzyl (2S, 4R) -4- (benzyloxy) -2-isopropenylpyrrolidine-1-carboxylate benzyl (2S, 4R) -4- (benzyloxy) -2- (1-hydroxy-1-methylethyl) pyrrolidine-1-carboxylate (2.22g, 6.01mmol) was dissolved in toluene (40mL) and triethylamine (10.0mL, 72mmol) under nitrogen. The mixture was cooled to-50 ℃ and thionyl chloride (0.44mL, 6.0mmol) was added dropwise. After stirring at-30 ℃ for 3 hours, the mixture was quenched by addition of water. The resulting mixture was extracted twice with ethyl acetate and the organic extracts were washed with brine and Na₂SO₄Drying, filtering and concentrating. The residue was chromatographed on silica gel (10% to 20% ethyl acetate/hexane) to give 1.10g (52%) of the olefin as a pale yellow oil. LC/MS (M + H)⁺m/z＝352.2；¹H NMR(CDCl₃)7.35(m，10H)，5.16(m，2H)，4.84(m，2H)，4.52(m，3H)，4.16(m，1H)，3.87(m，1H)，3.58(m，1H)，2.29(m，1H)，1.94(m，1H)，1.69(m，3H).

Step E

(2S, 4R) -4- (benzyloxy) -2-isopropylpyrrolidine benzyl (2S, 4R) -4- (benzyloxy) -2-isopropenylpyrrolidine-1-carboxylate (1.00g, 2.84mmol) was dissolved in ethanol (40mL), and 5% Pd-C (100mg) was added to the solution. The flask was purged with hydrogen and then shaken on Parr under 53psi hydrogen atmosphere for 17 hours. The reaction was then flushed with nitrogen, filtered through celite on a frit, and washed with methanol. The filtrate was concentrated and chromatographed on silica gel (1% triethylamine/10% methanol/89% ethyl acetate) to give the amine as a pale yellow oil, 0.53g (85%). LC/MS (M + H)⁺m/z＝220.2；¹H NMR(CDCl₃)7.33(m，5H)，4.49(m，2H)，4.12(m，1H)，3.19(dd，1H)，3.00(m，2H)，2.05(m，1H)，1.96(bs，1H)，1.49(m，2H)，1.00(d，3H)，0.91(d，3H).

Step F

N- {2- [ (2S, 4R) -4-benzyloxy-2-isopropylpyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. (2S, 4R) -4- (benzyloxy) -2-isopropylpyrrolidine (0.410g, 1.90mmol) was dissolved in dichloromethane (30mL) under nitrogen. (3-trifluoromethyl-benzoylamino) -acetic acid (0.462g, 1.90mmol) was added followed by EDC (0.394g, 2.06mmol) and the mixture was stirred at room temperature overnight. LC/MS revealed that the reaction was not complete. More (3-trifluoromethyl-benzoylamino) -acetic acid (0.12g, 0.48mmol) and more EDC (0.30g, 1.6mmol) were added and stirring at room temperature was continued for 3h, then 1.5 h under reflux. The mixture was chromatographed on silica gel, eluting with 30% ethyl acetate/hexane to give 0.66g (79%) of the coupled product as a colorless oil. LC/MS (M + H)⁺m/z＝449.2；¹H NMR(CDCl₃)8.03(m，1H)，7.76(m，1H)，7.58(m，2H)，7.34(m，5H)，4.52(m，2H)，4.03-4.34(m，4H)，3.65(m，1H)，3.48(m，1H)，2.54(m，1H)，2.12(m，1H)，1.92(m，1H)，0.92(d，3H)，0.77(d，3H).

Step G

N- {2- [ (2S, 4R) -4-hydroxy-2-isopropylpyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. N- {2- [ (2S, 4R) -4-benzyloxy-2-isopropylpyrrolidin-1-yl]-2-oxoEthyl } -3- (trifluoromethyl) benzamide (0.630g, 1.40mmol) was dissolved in methanol (60mL), and palladium hydroxide (90mg) was then added to the solution. The flask was purged with hydrogen and then stirred under hydrogen atmosphere using a balloon. After 3 hours, TLC indicated complete consumption of starting material. The reaction was then purged with nitrogen, filtered through celite on a frit, and washed with methanol. The filtrate was concentrated to give the desired alcohol as a white solid, 0.52g (100%). LC/MS (M + H)⁺m/z＝359.2；¹HNMR(CDCl₃)8.11(m，2H)，7.53-7.82(m，3H)，4.04-4.52(m，4H)，3.63(m，1H)，3.43(m，1H)，2.50(m，1H)，1.86-2.25(m，2H)，0.89(d，3H)，0.78(d，3H).

Step H

N- (2- { (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino]-2-isopropyl-pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The title compound was prepared from the above intermediate following the procedure described for example 59. Higher Rf isomer: LC/MS (M + H)⁺m/z＝533.3；¹H NMR(CD₃OD, bis-trifluoroacetate salt)8.66(m, 1H), 8.20(m, 3H), 7.94(m, 2H), 7.74(m, 1H), 7.59(m, 1H), 4.36(m, 2H), 4.06-4.27(m, 2H), 4.00(m, 1H), 3.63(m, 1H), 3.46(m, 1H), 2.63(m, 1H), 2.50(m, 1H), 2.34(m, 4H), 1.76-2.05(m, 5H), 0.96(d, 3H), 0.93(d, 3H); the lower Rf isomer: LC/MS (M + H)⁺m/z＝533.2；¹H NMR(CD₃OD, bis-trifluoroacetate) 8.66(m, 1H), 8.24(m, 3H), 7.96(m, 2H), 7.72(m, 2H), 4.00-4.42(m, 5H), 3.45(m, 2H), 2.65(m, 1H), 2.49(m, 1H), 2.22(m, 4H), 1.95(m, 5H), 0.96(d, 3H), 0.91(d, 3H).

The following examples 68-71 were prepared in a similar manner to example 67.

Example 68

N- {2- [ (2S,4S) -4- ({ 4-hydroxy-4- [5- (methoxymethyl) pyridin-2-yl)]Cyclohexyl } -amino) -2- (methoxymethyl) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. MS (M + H)⁺579.

Example 69

N- {2- [ (2S,4S) -4- [ (4- {5- [ (dimethylamino) methyl ] methyl]Pyridin-2-yl } -4-hydroxy-cyclohexyl) amino]-2- (methoxymethyl) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. MS (M + H)⁺592.

Example 70

N- {2- [ (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino]-2- (isopropoxy-methyl) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide. MS (M + H)⁺563.

Example 71

N- {2- [ (2S,4S) -4- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide. MS (M + H)613.3.

Example 72

N- (2- { (3S) -3- [ [ 4-hydroxy-4- (5-pyrazin-2-ylpyridin-2-yl) cyclohexyl ] (methyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. N- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrazin-2-ylpyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide (29.0mg, 0.051mmol) and 37% aqueous formaldehyde (21. mu.L, 0.26mmol) were dissolved in THF (1.0 mL). The mixture was evaporated to dryness. The residue was then dissolved in THF (1mL) and sodium triacetoxyborohydride (24mg, 0.11mmol) was added. After stirring at room temperature overnight, the mixture was purified by HPLC to give the title compound (5.9 mg). MS (M + H)583.3.

Example 73

N- (2- { (3S) -3- [ { 4-hydroxy-4- [5- (1, 3-)Azol-2-yl) pyridin-2-yl]Cyclohexyl } (methyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide. The reaction solution was purified by dissolving N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1, 3-)Azol-2-yl) pyridin-2-yl]Cyclohexyl } amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide (45mg, 0.081mmol) and 37% aqueous formaldehyde (30mg, 1.0mmol) were dissolved in methylene chloride (5.6 mL). The mixture was evaporated to dryness. The residue was then dissolved in THF (1mL) and sodium triacetoxyborohydride (38mg, 0.18mmol) was added. After stirring at room temperature overnight, the mixture was purified by HPLC to give the title compound (27 mg). MS (M + H)572.3.

Example 74

Step A

1- [3- (trifluoromethyl) phenyl]To a solution of piperidine-4-carboxylic acid methyl ester (2.0g, 14mmol), 1-bromo-3- (trifluoromethyl) benzene (1.5g, 6.8mmol) and potassium tert-butoxide (0.76g, 6.8mmol) in a mixed solvent of toluene (20mL) and DMF (4mL) under nitrogen was added [1, 1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride complex with dichloromethane (1: 1) (0.3g, 0.4 mmol). The mixture was heated in an oil bath at 130 ℃ overnight. After cooling to room temperature, the mixture was filtered through celite, diluting with EtOAc. The resulting solution was saturated NaHCO₃And (6) washing. The aqueous layer was extracted twice with EtOAc. The combined organic layers were dried (MgSO)₄) Concentrated and flash chromatographed using EtOAc/hexanes (20% to 40%) to give 0.90g of product. MS (M + H)288.2.

Step B

1- [3- (trifluoromethyl) phenyl ] piperidine-4-carboxylic acid methyl 1- [3- (trifluoromethyl) phenyl ] piperidine-4-carboxylate (0.9g, 3mmol) was treated with a mixture of 2M aqueous sodium hydroxide (10mL), THF (10mL) and methanol (10mL) at 50 ℃ for 1 hr. After neutralization (pH 3) with concentrated HCl, the solution was concentrated. The resulting residue was azeotroped with toluene three times to give the title compound, which was used in the next reaction without purification. MS (M + H)274.1.

Step C

[ (3S) -1- ({1- [3- (trifluoromethyl) phenyl]Piperidin-4-yl } carbonyl) pyrrolidin-3-yl](3S) -Pyrrolidin-3-ylcarbamic acid tert-butyl ester (0.65g, 3.5mmol), 1- [3- (trifluoromethyl) phenyl]Piperidine-4-carboxylic acid (0.80g, 2.9mmol), triethylamine (0.82mL, 5.8mmol) and benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (1.4g, 3.2mmol) were mixed in anhydrous methylene chloride (10 mL). After stirring overnight, the reaction mixture was diluted with EtOAc and saturated NaHCO₃And (6) washing. The aqueous layer was extracted three times with EtOAc. The combined organic layers were dried (MgSO)₄) Concentrated and flash chromatographed (20% EtOAc/hexanes to 40% EtOAc/hexanes) to give 0.975g of the desired product. MS (M + H)442.1.

Step D

(3S) -1- ({1- [3- (trifluoromethyl) phenyl ] piperidin-4-yl } carbonyl) pyrrolidin-3-amine bis (trifluoroacetate.) tert-butyl [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] carbamate (0.975g, 2.21mmol) was treated with trifluoroacetic acid (5mL) and methylene chloride (5mL) at room temperature for 1 hr. The solution was concentrated to give 1.75g of product, which was used in the next step without purification.

Step E

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl]Piperidin-4-yl } carbonyl) pyrrolidin-3-yl]Amino } cyclohexanols (3S) -1- ({1- [3- (trifluoromethyl) phenyl)]Piperidin-4-yl } carbonyl) pyrrolidin-3-amine bis (trifluoroacetate) (110mg, 0.20mmol), 4-hydroxy-4-pyridin-2-yl-cyclohexanone (45mg, 0.24mmol), triethylamine (0.082mL, 0.59mmol), and sodium triacetoxyborohydride (83mg, 0.39mmol) were combined in dichloromethane (6 mL). After stirring overnight, the reaction mixture was diluted with EtOAc and saturated Na₂CO₃And (6) washing. The aqueous layer was extracted three times with EtOAc. The combined organic layers were dried (MgSO)₄) Concentrated and purified over silica gel column (EtOAc to 1% Et)₃N/EtOAc to 5% Et₃N/EtOAc) to give the title compound. LCMS (M + H) ═ 517.2.

Example 75

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol the title compound was prepared in a manner analogous to that described in example 74. MS (M + H)595.2.

Example 76

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol the title compound was prepared in a manner analogous to that described for example 74. MS (M + H)581.2.

Example 77

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] azetidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol the title compound was prepared in a manner analogous to that described for example 74. MS (M + H)567.2.

Example 78

Step A

1- [ (benzyloxy) carbonyl]Triethylamine (8.1mL, 58mmol) was added to a solution of piperidine-4-carboxylic acid (5g, 40mmol) and benzyl chloroformate (7.9g, 46mmol) in dichloromethane (100mL) in an ice-water bath. After stirring overnight, the solution was washed with concentrated HCl and brine, Na₂SO₄Drying and concentrating. Chromatography on silica gel afforded the title compound (10g) as an oil. MS (M + H)264.2.

Step B

4- ({ (3S) -3- [ (tert-Butoxycarbonyl) amino]Pyrrolidin-1-yl } carbonyl) piperidine-1-carboxylic acid benzyl ester 1- [ (benzyloxy) carbonyl]A mixture of piperidine-4-carboxylic acid (5g, 20mmol), (3S) -pyrrolidin-3-ylcarbamic acid tert-butyl ester (3.9g, 21mmol), benzotriazol-1-yloxytris (dimethylamino) hexafluorophosphate (9.2g, 21mmol) and triethylamine (3.8g, 38mmol) in dichloromethane (100mL) was stirred at room temperatureAnd (4) at night. The reaction solution was washed with water and Na₂SO₄Drying and concentrating. The residue was chromatographed on silica gel to give 7.5g of product. MS (M + H)432.2.

Step C

[ (3S) -1- (piperidin-4-ylcarbonyl) pyrrolidin-3-yl ] carbamic acid tert-butyl ester A mixture of benzyl 4- ({ (3S) -3- [ (tert-butoxycarbonyl) amino ] pyrrolidin-1-yl } carbonyl) piperidine-1-carboxylate (7.5g, 17mmol) and palladium on charcoal (800mg, 8mmol) in methanol (100mL) was shaken under 50psi of hydrogen overnight. The mixture was filtered through celite and the filtrate was concentrated to give 5.1g of product as a white solid. MS (M + H)298.2.

Step D

[ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl)]Piperidin-4-yl } carbonyl) pyrrolidin-3-yl](ii) carbamic acid tert-butyl ester 2-chloro-6- (trifluoromethyl) pyridine (1.8g, 9.9mmol), [ (3S) -1- (piperidin-4-ylcarbonyl) pyrrolidin-3-yl]A solution of tert-butyl carbamate (2.97g, 10.0mmol) and triethylamine (4.1mL, 30mmol) in DMF (50mL) was heated at 100 deg.C for 4 hr. After cooling down, ethyl acetate was added. The resulting solution was washed several times with brine and over Na₂SO₄Drying and concentrating. The residue was chromatographed on silica gel to give the title compound (1.3g) as a yellow solid. MS (M + H)443.2.

Step E

(3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl]Piperidin-4-yl } carbonylYl) pyrrolidin-3-amine. Reacting [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl)]Piperidin-4-yl } carbonyl) pyrrolidin-3-yl]Tert-butyl carbamate (1.3g, 2.9mmol) in 4M HCl in 1, 4-bisAlkane solution (10 mL). After stirring at room temperature for 1hr, the solution was concentrated to give the desired product as HCl salt (0.6 g). MS (M + H)343.1.

Step F

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-amine (40mg, 0.1mmol), 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexanone (47mg, 0.18mmol), sodium triacetoxyborohydride (50mg, 0.23mmol) and triethylamine (35mg, 0.35mmol) of dichloromethane (10mL) was stirred at room temperature overnight. The reaction mixture was passed through a pad of silica gel. The filtrate was concentrated and purified by HPLC to give cis-and trans-isomers. MS (M + H)596.2 for both isomers.

The following examples were prepared in a similar manner to example 78.

Example 79

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)518.2.

Example 80

1- (6-pyrimidin-2-ylpyridin-3-yl) -4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyrimidin-4-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)597.3.

Example 81

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyrimidin-4-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)51.9.2.

Example 82

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)518.2.

Example 83

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)596.2.

Example 84

1-[5-(1，3-Azol-2-yl) pyridin-2-yl]-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl]Piperidin-4-yl } carbonyl) pyrrolidin-3-yl]Amino } cyclohexanol MS (M + H)584.2.

Example 85

1- (5-pyrazin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)596.2.

Example 86

1- (5-methylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)532.2.

Example 87

1- (3,3' -bipyridin-6-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)595.3.

Example 88

1- (3,4' -bipyridin-6-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)595.3.

Example 89

1- (5-methoxypyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)548.2.

Example 90

1- [5- (methoxymethyl) pyridin-2-yl ] -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)562.2.

Example 91

6- (1-hydroxy-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexyl) nicotinamide. MS (M + H)561.3.

Example 92

6- (1-hydroxy-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexyl) -N-methylnicotinamide. MS (M + H)575.3.

Example 93

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyrimidin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)597.4.

Example 94

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)582.2.

Example 95

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [5- (trifluoromethyl) pyridin-2-yl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)582.3.

Example 96

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyrimidin-2-yl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)583.3.

Example 97

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({ (3R) -1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)596.4.

Example 98

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({ (3S) -1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)596.4.

Example 99

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] azetidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol MS (M + H)568.1.

Example 100

Step A

1- [4- (trifluoromethyl) pyridin-2-yl]To a solution of methyl 1H-imidazole-4-carboxylate (417mg, 3.3mmol) in DMF (10mL) was added sodium hydride (130mg, 3.3 mmol). After stirring at room temperature for 1hr, 2-chloro-4- (trifluoromethyl) pyridine (500mg, 2.8mmol) was added. The mixture was stirred at 80 ℃ overnight. After cooling to room temperature, ethyl acetate was added. The solution was washed several times with brine and dried (MgSO)₄) And (4) concentrating. Chromatography on silica eluting with EtOAc/hexanes (1: 1) afforded the title compound (120 mg). MS (M + H)272.1.

Step B

1- [4- (trifluoromethyl) pyridin-2-yl ] -1H-imidazole-4-carboxylic acid to a solution of methyl 1- [4- (trifluoromethyl) pyridin-2-yl ] -1H-imidazole-4-carboxylate (120mg, 0.44mmol) in methanol (2.5mL) was added 5M aqueous sodium hydroxide (2.5mL) and the mixture was stirred at room temperature for 1 hr. After removal of methanol under vacuum, the resulting solution was acidified (pH 5) with concentrated HCl and concentrated. The residue was dissolved in acetone and insoluble material was filtered off. The filtrate was evaporated to give the title compound (120 mg). MS (M + H)258.2.

Step C

[ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] -1H-imidazol-4-yl } carbonyl) pyrrolidin-3-yl ] carbamic acid tert-butyl ester to a solution of 1- [4- (trifluoromethyl) pyridin-2-yl ] -1H-imidazole-4-carboxylic acid (120mg, 0.47mmol) and tert-butyl (3S) -pyrrolidin-3-ylcarbamate (87mg, 0.47mmol) in DMF (3mL) was added benzotriazol-1-yloxytris (dimethylamino) hexafluorophosphate (210mg, 0.47mmol) followed by triethylamine (0.20mL, 1.4 mmol). The reaction was stirred at room temperature overnight and purified by HPLC to give the title compound. MS (M + H)426.3.

Step D

1- ({ (3S) -1- [4- (trifluoromethyl) pyridin-2-yl]-1H-imidazol-4-yl } carbonyl) pyrrolidin-3-amine. To [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl)]-1H-imidazol-4-yl } carbonyl) pyrrolidin-3-yl]Tert-butyl carbamate (120mg, 0.28mmol) in methanol (2mL) was added 4.0M HCl in 1, 4-bisAlkane solution (3.0 mL). After stirring for 0.5hr, the solution was concentrated under vacuum to give the title compound. MS (M + H)326.2.

Step E

1- (6-pyrimidin-2-ylpyridin-3-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl]-1H-imidazol-4-yl } carbonyl) pyrrolidin-3-yl]Amino } cyclohexanol to (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl]-1H-imidazol-4-yl } carbonyl) pyrrolidin-3-amine (50mg, 0.15mmol) and 4-hydroxy-4- (6-pyrimidin-2-ylpyridin-3-yl) cyclohexanone (41mg, 0.15mmol) in methylene chloride (3mL) was added sodium triacetoxyborohydride (36mg, 0.17mmol) followed by triethylamine (0.086mL, 0.61 mmol). After stirring at room temperature for 2hr, EtOAc (50mL) was added. The solution is treated with NaHCO₃The solution was washed with water and dried (MgSO)₄) And (4) concentrating. Purification by HPLC gave two isomers. MS (M + H)579.3 for both isomers.

Example 101

Step A

2-methyl-4- (trifluoromethyl) pyridine 1-oxide to a solution of 2-methyl-4- (trifluoromethyl) pyridine (3.9g, 24mmol) in methylene chloride (50mL) was added m-chloroperbenzoic acid (7.0g, 31 mmol). After stirring overnight at room temperature, the solution was washed with 50mL of 1N NaOH. The aqueous phase was back-extracted with methylene chloride. The organic phases were combined and passed over Na₂SO₄Dried and concentrated in vacuo to afford the title compound. MS (M + H)178.1.

Step B

Acetic acid [4- (trifluoromethyl) pyridin-2-yl]Methyl ester 2-methyl-4- (trifluoromethyl) pyridine 1-oxide (4.0g, 22mmol) was added to acetic anhydride (12mL) at 120 ℃. The mixture was refluxed for 1 hr. 10mL of ethanol was carefully added thereto. Refluxing is continued for 10 min. Pouring the mixtureIn ice, with NaHCO₃Neutralization with Et₂And (4) extracting. The organic layer was dried (MgSO₄) And (4) concentrating. Chromatography on silica gel (5: 2 hexane/EtOAc) afforded the product (3.4g) as a brown oil. MS (M + H)220.1.

Step C

[4- (trifluoromethyl) pyridin-2-yl]To acetic acid [4- (trifluoromethyl) pyridin-2-yl]A solution of methyl ester (1.0g, 3.2mmol) in methanol (10mL) was added 1.0M aqueous sodium hydroxide (10 mL). After stirring at room temperature overnight, the solution was diluted with 20mL of water and extracted twice with EtOAc. The combined organic layers were dried (MgSO)₄) And concentrated under vacuum. Chromatography on silica gel eluting with hexane/EtOAc (1: 1) afforded the title compound (0.34g) as a clear oil. MS (M + H)178.1.

Step D

{ [4- (trifluoromethyl) pyridin-2-yl]Methoxy } acetic acid to [4- (trifluoromethyl) pyridin-2-yl]Methanol (340mg, 1.9mmol) in DMF (10mL) was added sodium hydride (150mg, 3.8 mmol). After stirring at room temperature for 5min, bromoacetic acid 1, 1-dimethylethyl ester (0.28mL, 1.9mmol) was added. Stirring was continued at room temperature for 1 hr. Water (20mL) was added and the resulting solution was extracted with EtOAc. The aqueous layer was neutralized with HCl to pH 5 and extracted twice with EtOAc. The combined organic layers were dried (MgSO)₄) Concentration under vacuum afforded the title compound, which was used in the next reaction without purification. MS (M + H)292.2.

Step E

To a solution of { [4- (trifluoromethyl) pyridin-2-yl ] methoxy } acetyl) pyrrolidin-3-yl ] carbamic acid tert-butyl ester (450mg, 1.9mmol) and tert-butyl (3S) -pyrrolidin-3-ylcarbamate (360mg, 1.9mmol) in DMF (10mL) was added benzotriazol-1-yloxytris (dimethylamino) hexafluorophosphate (880mg, 2.0mmol) followed by triethylamine (0.80mL, 5.7 mmol). After stirring overnight at room temperature, ethyl acetate was added. The solution was washed with 1N NaOH and water. Purification on a silica gel column eluting with EtOAc provided the title compound (300mg) as a clear oil. MS (M + H)404.3.

Step F

1- ({ [4- (trifluoromethyl) pyridin-2-yl)]Methoxy } acetyl) pyrrolidin-3-amine. To [ (3S) -1- ({ [4- (trifluoromethyl) pyridin-2-yl)]Methoxy } acetyl) pyrrolidin-3-yl]Tert-butyl carbamate (300mg, 0.74mmol) in methanol (3mL) was added 4.0M HCl in 1, 4-bisAlkane solution (6 mL). After stirring at room temperature for 0.5hr, the solution was concentrated under vacuum to give the title compound. MS (M + H)304.2.

Step G

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({ [4- (trifluoromethyl) pyridin-2-yl]Methoxy } acetyl) pyrrolidin-3-yl]Amino } cyclohexanol to (3S) -1- ({ [4- (trifluoromethyl) pyridin-2-yl]Methoxy } acetyl) pyrrolidin-3-amine (47mg, 0.15mmol) and 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexanone (41mg, 0.15mmol) in methanol (2mL) and isopropanolA solution of propanol (2mL) was added sodium triacetoxyborohydride (36mg, 0.17 mmol). After stirring at room temperature overnight, EtOAc was added. The solution is treated with NaHCO₃The solution was washed with water and dried (MgSO)₄) And (4) concentrating. Purification by HPLC gave the title compound as two isomers. MS (M + H)557.2 for both isomers.

Example 102

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({ [4- (trifluoromethyl) 2-phenyl ] methoxy } acetyl) pyrrolidin-3-yl ] amino } cyclohexanol the title compound was prepared in a similar manner to example 101. MS (M + H)556.3.

Pharmaceutical use of the compounds of the invention

The ability of the novel compounds of the present invention to antagonize CCR2 function can be determined using a suitable screening method (e.g., a high throughput assay). For example, agents can be tested in extracellular acidification assays, calcium flux assays, ligand binding assays, or chemotaxis assays (see, e.g., Hesselgesser et al, J biol. chem.273 (25): 15687-.

In actual assays, an isolated or recombinantly derived CCR2 protein is used that has at least one property, activity or functional characteristic of a mammalian CCR2 protein. Specific properties may be binding properties (e.g. with ligands or inhibitors), signalling activity (e.g. activation of mammalian G-protein, cytosolic free calcium concentration [ Ca ]⁺⁺]induction of a rapid transient increase in i), cellular response functions (e.g., stimulation of chemotaxis or inflammatory mediator release by leukocytes), and the like.

In one embodiment, a composition comprising a CCR2 protein or variant thereof is maintained under conditions suitable for binding. The CCR2 receptor is contacted with the compound to be tested and binding is detected or measured.

In an alternative embodiment, the assay is a cell-based assay using cells stably or transiently transfected with a vector or expression cassette having a nucleic acid sequence encoding a CCR2 receptor. The cells are maintained under conditions suitable for expression of the receptor and contacted with the agent under conditions suitable for binding to occur. Binding can be detected using standard techniques, e.g., the degree of binding can be determined relative to a suitable control. Also, a cell fraction containing receptors, such as a membrane fraction, may be used instead of whole cells.

Binding or complex formation may be detected directly or indirectly. For example, the drug may be labeled with a suitable label (e.g., fluorescent label, tag, isotopic label, enzymatic label, etc.) and binding may be determined by detecting the label. Specific and/or competitive binding can be assessed by competition or displacement studies using unlabeled drug or ligand as a competitor.

CCR2 antagonist activity of a test drug (e.g., a 3-cycloalkylaminopyrrolidine compound of formula I or II of the present invention) can be reported as the concentration of inhibitor required to inhibit 50% specific binding (IC) in a receptor binding assay₅₀Value), in said assay, use¹²⁵Peripheral Blood Mononuclear Cells (PBMC) prepared from normal human whole blood via density gradient centrifugation using I-labeled MCP-1 as a ligand. Specific binding is preferably defined as total binding (e.g., total cpm on the filter) minus non-specific binding. Non-specific binding is defined as the amount of cpm still detected in the presence of excess unlabeled competitor (e.g., MCP-1).

The human PBMCs described above may be used in a suitable binding assay. For example, 200,000 to 500,000 cells can be contacted with 0.1 to 0.2nM¹²⁵I-labeled MCP-1 incubation with or without unlabeled competitor (10nM MCP-1) or varying concentrations of test compound.¹²⁵I-labeled MCP-1 may be prepared by a suitable method or purchased from a supplier (Perkin Elme)r, Boston MA). The binding reaction can be performed for 30min at room temperature in 50 to 250 μ L binding buffer consisting of 1M HEPES pH 7.2 and 0.1% BSA (bovine serum albumin). The binding reaction can be stopped by harvesting the membrane by rapid filtration through a glass fiber filter (Perkin Elmer) pre-soaked in 0.3% polyethyleneimine or Phosphate Buffered Saline (PBS). The filter can be washed with approximately 600. mu.L of binding buffer containing 0.5M NaCl or PBS, then dried, and the amount of bound radioactivity can be determined by counting on a Gamma counter (Perkin Elmer).

The ability of a compound to antagonize CCR2 function can also be determined in a leukocyte chemotaxis assay using suitable cells. Suitable cells include, for example, cell lines, recombinant cells or isolated cells that express CCR2 and undergo CCR2 ligand-induced (e.g., MCP-1) chemotaxis. The assay employs human peripheral blood mononuclear cells, and a modified Boyden chamber (Neuro Probe). 500,000 cells were incubated in serum-free DMEM medium (InVitrogen) with or without inhibitors and warmed to 37 ℃. The chemotaxis chamber (Neuro Probe) was also preheated. To all wells in the bottom chamber, 400. mu.L of warm 10nM MCP-1 was added, but in the negative control DMEM was added. An 8 micron membrane filter (Neuro Probe) was placed on top and the lid was closed. Cells are then added to the well in the lid, communicating with the small hole below the filter. The entire chamber was incubated at 37 ℃ under 5% CO2 for 30 minutes. The cells were then aspirated, the lid opened, and the filter carefully removed. The filter top was washed three times with PBS and the bottom was stationary. The filters were air dried and stained with Wright Geimsa stain (Sigma). The filters were counted by microscopy. Negative control wells served as background and were subtracted from all values. The efficacy of the antagonist can be determined by comparing the number of cells that migrated to the bottom chamber in wells containing the antagonist to the number of cells that migrated to the bottom chamber in MCP-1 control wells.

IC of Compounds of the invention when Using a binding assay protocol₅₀In the range of about 0.01 to about 500 (nM). IC of Compounds of the invention in chemotaxis assays₅₀In the range of about 1 to about 3000 (nM).

A method of modulating chemokine receptor activity comprising contacting said chemokine receptor with a compound of claim. Chemokine receptors to which the compounds of the invention bind and/or modulate include any chemokine receptor. In some embodiments, the chemokine receptor belongs to the CC family of chemokines, including, for example, CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, and CCR 8. In some embodiments, the chemokine receptor is CCR 2. In some embodiments, the chemokine receptor is CCR 5. In some embodiments, the chemokine receptor binds and/or modulates CCR2 and CCR 5.

The term "contacting" as used herein means bringing the indicated entities together in an in vitro system or in an in vivo system. For example, "contacting" a chemokine receptor with a compound of the invention includes administration of a compound of the invention to an individual or patient, e.g., a human, having a chemokine receptor, and introducing a compound of the invention, e.g., into a sample containing cells or purified preparations containing a chemokine receptor.

The compounds of the invention may be selective. By "selective" is meant that the compound, when binding or inhibiting a chemokine receptor, has greater affinity or potency, respectively, than at least one other chemokine receptor, or preferably, all other chemokine receptors in the same class (e.g., all other CC type receptors). In some embodiments, the compounds of the invention have binding or inhibitory selectivity for CCR2 or CCR5 over any other chemokine receptor. The selectivity can be at least about 10 fold, at least about 20 fold, at least about 50 fold, at least about 100 fold, at least about 200 fold, at least about 500 fold, or at least about 1000 fold. Binding affinity and inhibitor potency can be measured according to methods customary in the art, for example according to the assays provided herein.

The invention further provides methods of treating a chemokine receptor-associated disease or disorder in an individual (e.g., patient) by administering to an individual in need of such treatment a therapeutically effective amount or dose of a compound of the invention or a pharmaceutical composition thereof. Chemokine receptor-associated diseases can include any disease, disorder or condition that is directly or indirectly linked to the expression or activity of a chemokine receptor. Chemokine receptor-associated diseases can also include any disease, disorder or condition that can be prevented, ameliorated or cured by modulation of chemokine receptor activity. A chemokine receptor-associated disease can further include any disease, disorder or condition characterized by the binding of an infectious agent, such as a virus or viral protein, to a chemokine receptor. In some embodiments, the chemokine receptor-associated disease is a CCR 5-associated disease, such as HIV infection.

As used herein, the terms "individual" or "patient" are used interchangeably and refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans. The compounds of the invention can be administered to a mammal, such as a human, but can also be other mammals, such as animals in need of veterinary treatment, e.g., domestic animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, sheep, pigs, horses, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). The mammal to be treated in the method of the invention is a mammal in need of modulation of chemokine receptor activity, and may be male or female. The term modulation is intended to encompass antagonism (e.g., inhibition), agonism, partial antagonism and/or partial agonism. In some embodiments, the compounds of the present invention are antagonists (e.g., inhibitors) of chemokine receptors.

In this specification, the term "therapeutically effective amount" means that amount of the subject compound that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician.

The compounds of the present invention are administered in therapeutically effective amounts to treat certain diseases, such as rheumatoid arthritis. A therapeutically effective amount of the compound is effective in treating a subject suffering from and/or having aberrant leukocyte recruitment and/or activationAn amount that results in inhibition of one or more processes mediated by binding of a chemokine to a receptor, e.g., CCR2, in a subject with the disease in question. Typical examples of such processes include leukocyte migration, integrin activation, intracellular free calcium concentration [ Ca [ ]²⁺]A transient increase in i and a granular release of proinflammatory mediators. Alternatively, a therapeutically effective amount of a compound is that amount necessary to achieve the desired therapeutic and/or prophylactic effect, e.g., an amount that results in the prevention or reduction of symptoms associated with a disease associated with aberrant leukocyte recruitment and/or activation.

Other diseases or conditions of human or other animal species that may be treated with the inhibitors or modulators of chemokine receptor function of this invention include, but are not limited to: inflammatory or allergic diseases and conditions, including respiratory allergic diseases, such as asthma, allergic rhinitis, allergic lung disease, allergic pneumonia, eosinophilic cellulitis (e.g., Welss syndrome), eosinophilic pneumonia (e.g., Loeffler's syndrome, chronic eosinophilic pneumonia), eosinophilic polymyositis (e.g., Shulman's syndrome), delayed-type hypersensitivity, Interstitial Lung Disease (ILD) (e.g., idiopathic pulmonary fibrosis, or ILD associated with rheumatoid arthritis, systemic lupus erythematosus, ankylosing spondylitis, systemic sclerosis, Sjogren's syndrome, polymyositis, or dermatomyositis); systemic anaphylaxis or anaphylaxis, drug allergy (e.g., to penicillin, cephalosporin), eosinophilic myalgia syndrome caused by intake of contaminated tryptophan, insect bite allergy; autoimmune diseases, such as rheumatoid arthritis, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, myasthenia gravis, juvenile onset diabetes; glomerulonephritis, autoimmune thyroiditis, Behcet's disease, graft rejection (e.g., in transplantation), including allograft rejection or graft-versus-host disease; inflammatory bowel diseases, such as crohn's disease and ulcerative colitis; spondyloarthropathy; scleroderma; psoriasis (including T cell mediated psoriasis) and inflammatory dermatoses such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis (e.g., necrotizing, cutaneous, and allergic vasculitis); eosinophilic myositis, eosinophilic fasciitis; cancer with leukocyte infiltration of the skin or organs. Other diseases or conditions where inhibition of an undesirable inflammatory response is desirable include, but are not limited to, reperfusion injury, atherosclerosis, restenosis, certain hematologic malignancies, cytokine-induced toxicity (e.g., septic shock, endotoxic shock), polymyositis, dermatomyositis.

In some embodiments, chemokine receptor-associated diseases, disorders and conditions include inflammation and inflammatory diseases, immune disorders, cancer and viral infections. Examples of inflammatory diseases include diseases having an inflammatory component, such as asthma, allergic rhinitis, restenosis, atherosclerosis, multiple sclerosis, Crohn's disease, ulcerative colitis, allergic lung disease, neuropathic pain, hypersensitivity pneumonitis, eosinophilic pneumonia, delayed-type hypersensitivity, asthma, Interstitial Lung Disease (ILD) (e.g., idiopathic pulmonary fibrosis, or ILD associated with rheumatoid arthritis, systemic lupus erythematosus, ankylosing spondylitis, systemic sclerosis, Sjogren's syndrome, polymyositis or dermatomyositis), ocular disorders (e.g., retinal neurodegeneration, choroidal neovascularization, etc.), obesity, and the like. Examples of immune disorders include rheumatoid arthritis, psoriatic arthritis, systemic lupus erythematosus, myasthenia gravis, diabetes (e.g., juvenile onset diabetes), insulin resistance; glomerulonephritis, autoimmune thyroiditis, organ transplant rejection (including allograft rejection and graft-versus-host disease). Examples of cancers include breast cancer, ovarian cancer, multiple myeloma, and the like, which are characterized by infiltration of macrophages (e.g., tumor-associated macrophages, TAMs) into the tumor or diseased tissue. Examples of viral infections include HIV infection.

One or more additional pharmaceutical ingredients, such as antibodies, anti-inflammatory agents, immunosuppressive agents, chemotherapeutic agents, may be used in combination with the compounds of the present invention for the treatment of a chemokine receptor associated disease, disorder or condition. These agents may be combined with the compounds of the present invention in a single dosage form, or the agents may be administered as separate dosage forms, either simultaneously or sequentially.

One or more additional pharmaceutical ingredients, such as antiviral agents, antibodies, anti-inflammatory agents, and/or immunosuppressive agents, may be used in combination with the compounds of the present invention for the treatment of a chemokine receptor-associated disease, disorder, or condition. These agents may be combined with the compounds of the present invention in a single dosage form, or the agents may be administered as separate dosage forms, either simultaneously or sequentially.

Antiviral agents suitable for use in combination with the compounds of the present invention may comprise nucleoside and Nucleotide Reverse Transcriptase Inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitors and other antiviral agents.

Examples of suitable NRTIs include zidovudine (AZT), didanosine (ddl), zalcitabine (ddC), stavudine (D4T), lamivudine (3TC), abacavir (1592U89), adefovir dipivoxil [ bis (POM) -PMEA ], lobecavir (BMS-180194), BCH-10652, emtricitabine [ (-) -FTC ], β -L-FD4 (also known as β -L-D4C and β -L-2 ', 3' -dicloxy-5-fluoro-cydene), DAPD ((-) - β -D-2, 6, -diamino-purine dioxolane), and lodenosine (FddA).

Typical suitable NNRTIs include nevirapine (BI-RG-587), delaviradine (BHAP, U-90152), efavirenz (DMP-266), PNU-142721, AG-1549, MKC-442(1- (ethoxy-methyl) -5- (1-methylethyl) -6- (phenylmethyl) - (2, 4(1H, 3H) -pyrimidinedione), and (+) -calanolide A (NSC-675451) and B.

Typical suitable protease inhibitors include saquinavir (Ro 31-8959), ritonavir (ABT-538), indinavir (MK-639), nelfnavir (AG-1343), amprenavir (141W94), lacinavir (BMS-234475), DMP-450, BMS-2322623, ABT-378, and AG-1549.

Other antivirals include hydroxyurea, ribavirin, IL-2, IL-12, pentafuside and Yissum Project No. 11607.

In some embodiments, anti-inflammatory or analgesic agents contemplated for use in combination with the compounds of the present invention may comprise, for example, opioid agonists, lipoxygenase inhibitors (e.g., 5-lipoxygenase inhibitors), cyclooxygenase inhibitors (e.g., cyclooxygenase-2 inhibitors), interleukin inhibitors (e.g., interleukin-I inhibitors), NNMA antagonists, nitric oxide inhibitors or nitric oxide synthesis inhibitors, non-steroidal anti-inflammatory agents, or cytokine-inhibitory anti-inflammatory agents, such as acetaminophen, aspirin, codeine, fentanyl, ibuprofen, indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam, steroidal analgesic, sufentanil, sulindac, tenidap, and the like. Similarly, the compounds of the present invention may be administered with: a pain relieving agent; fortifiers, such as caffeine; h2-antagonists, such as simethicone, aluminum or magnesium hydroxide; decongestants such as phenylephrine, phenylpropanolamine, pseudoephedrine, oxymetazoline, ephinephrine, naphazoline, xylometazoline, propylhexedrine, or levo-desoxyephedrine; antifitutive, such as codeine, hydrocodone, caramiphene, pentoxyverine or dextromethorphan (dextramethorphan); a diuretic; and sedating or non-sedating antihistamines.

In some embodiments, it is contemplated that pharmaceutical compositions that may be used in combination with the compounds of the present invention may comprise (a) a VLA-4 antagonist, such as those described in US 5,510,332, WO 95/15973, WO 96/01644, WO 96/06108, WO 96/20216, WO 96/229661, WO96/31206, WO 96/4078, WO 97/030941, WO 97/022897, WO98/426567, WO 98/53814, WO 98/53817, WO 98/538185, WO 98/54207, and WO 98/58902; (b) steroids such as beclomethasone, methylprednisolone, betamethasone, prednisone, dexamethasone, and hydrocortisone; (c) immunosuppressants such as cyclosporin, tacrolimus, rapamycin, and other FK506 type immunosuppressants; (d) antihistamines (H1-histamine antagonists), e.g. brompheniramine, chlorpheniramine, dexchlorpheniramine, triprolidine, clemastine, diphenhydramine, diphenyllene, tripelennamine, hydroxyzine, methdilazine, promethamine, alimemazine, alazineZatadine, cyproheptadine, antazoline, pheniramine, pyrilamine, asimilole, terfenadine, loratadine, cetirizine, fexofenadine (fexofenadine), descarboethoxyloratadine (deserboethoxy loratadine), and the like; (e) non-steroidal anti-asthmatics such as terbutaline, metaproterenol, fenoterol, isotaline, salbutamol, bitolterol, pirbuterol, theophylline, cromolyn sodium, atropine, ipratropium bromide, leukotriene antagonists (e.g., zafirlukast, montelukast, pranlukast, iralukast, pobilukast, SKB-106, 203), leukotriene biosynthesis inhibitors (e.g., zileuton, BAY-1005); (f) non-steroidal anti-inflammatory agents (NSAIDs), e.g. propionic acid derivatives (e.g. alminoprofen, benzene)Loxfen, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen, pranoprofen, suprofen, tiaprofenic acid, and thioprofenLoxacin), acetic acid derivatives (e.g. indomethacin, acemetacin, alclofenac, clidanac, diclofenac, fenclofenac, fentiazac, furofenac, ibufenac, isoxepac, oxyphenbutac (oxipinac), sulindac, thiopinac, tolmetin, zidometacin and zomepic), fenamic acid derivatives (flufenamic acid, meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid), biphenylcarboxylic acid derivatives (diflunisal and flufenisal), oxicams (isoxicam, piroxicam, sudoxicam and tenoxicam), salicylic acids (acetylsalicylic acid, sulfasalazine) and pyrazolones (azapropazone, bezoperyl, feprazan, mofebuzzone, oxybutylazone, phenylbutazone); (g) cyclooxygenase-2 (COX-2) inhibitors; (h) phosphodiesterase type IV (PDE-IV) inhibitors; (i) antagonists of other chemokine receptors, especially CXCR-4, CCR1, CCR2, CCR3 and CCR 5; (j) cholesterol lowering agents, e.g. HMG-CoA reductase inhibitors (lovastatin, simvastatin)Pravastatin, fluvastatin, atorvastatin and other statins), chelating agents (cholestyramine and colestipol), nicotinic acid, fenofibric acid derivatives (gemfibrozil, clofibrate, fenofibrate and bezafibrate) and probucol; (k) antidiabetic agents such as insulin, sulfonylureas, biguanides (metformin), u-glucosidase inhibitors (acarbose), and glitazones (troglitazone and pioglitazone); (l) Interferon beta preparations (interferon beta-1 alpha, interferon beta-1 beta); (m) other compounds, such as aminosalicylic acid, antimetabolites, such as azathioprine and 6-mercaptopurine, and cytotoxic cancer chemotherapeutic agents. The weight ratio of the compound of the present invention to the second active ingredient may vary depending on the effective dose of each ingredient.

Rheumatoid Arthritis (RA) patients treated aggressively with disease mitigators (methotrexate, antimalarials, gold, penicillamine, sulfasalazine, dapsone, leflunomide, or biologics) can achieve varying degrees of disease control, including complete regression. These clinical responses are associated with an increase in standardized disease activity scores, specifically ACR criteria, which include: pain, function, number of tender joints, number of swollen joints, patient population assessment, physician population assessment, laboratory measurements of inflammation (CRP and ESR), and radiological assessment of joint structural damage. Current disease modifying drugs (DMARDs) require continuous administration to maintain optimal benefit. Long-term administration of these drugs is associated with significant toxicity and impaired host defense. In addition, patients often become refractory to a particular therapy, requiring an alternative regimen. For these reasons, novel, effective therapies that allow withdrawal of standard DMARDs would be a clinically significant advance.

Patients who have achieved clinical regression of disease who have significantly responded to anti-TNF therapy (infliximab, etanercept, adalimumab), anti-IL-1 therapy (kinaret), or other disease modifying anti-rheumatic drugs (DMARDs) including but not limited to methotrexate, cyclosporine, gold salts, antimalarials, penicillamine, or leflunomide may be treated with agents that inhibit CCR2 expression and/or activity, such as nucleic acids (e.g., antisense or siRNA molecules), proteins (e.g., anti-CCR 2 antibodies), small molecule inhibitors (e.g., the compounds disclosed herein and other chemokine receptor inhibitors known in the art).

In some embodiments, the agent that inhibits CCR2 expression and/or activity is a small molecule CCR2 inhibitor (or antagonist). The CCR2 antagonist may be administered orally four times a day (q.d.) or twice a day (b.i.d.) at a dose of no more than about 500mg per day. Patients may be withdrawn or dose reduced from current therapy and adhere to treatment with a CCR2 antagonist. Treatment of patients with a combination of a CCR2 antagonist and current therapy can be performed, for example, for about one to about two days, followed by discontinuation or dose reduction of the DMARD and continued CCR2 antagonist treatment.

The advantages of replacing traditional DMARDs with CCR2 antagonists are numerous. Traditional DMARDs have severe cumulative dose-limiting side effects, most commonly liver damage and immunosuppressive effects. CCR2 antagonism is expected to have improved long-term safety and will not have similar immunosuppressive tendencies associated with traditional DMARDs. In addition, the half-life of biologies is typically days or weeks, which is a problem when dealing with adverse reactions. The half-life of orally bioavailable CCR2 antagonists is expected to be on the order of hours, and thus the risk of sustained exposure to the drug following an adverse event is very minimal compared to biological agents. Furthermore, current biological agents (infliximab, etanercept, adalimumab, kinaret) are typically administered intravenously or subcutaneously, requiring either physician administration or patient self-injection. This gives rise to the possibility of infusion reactions or injection site reactions. These problems can be avoided by using an orally administered CCR2 antagonist.

The compounds of the invention can be administered in oral dosage forms, such as tablets, capsules (each including sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions. They may also be administered intravenously (bolus or infusion), intraperitoneally, subcutaneously, or intramuscularly, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts. They may be administered alone or in combination with a pharmaceutical carrier, which is selected with regard to the chosen route of administration and standard pharmaceutical practice.

The dosage regimen for the compounds of the invention will, of course, vary depending upon known factors, such as the pharmacodynamic characteristics of the particular ingredient and its mode and route of administration; metabolic stability, rate of excretion, drug combination and length of action of the compound; the recipient's breed, age, sex, health, medical condition, and weight; the nature and extent of the symptoms; the kind of concurrent therapy; the frequency of treatment; the specific route of administration; renal and hepatic function of the patient; and the desired effect. A physician or veterinarian can determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the particular disorder for which treatment is indicated.

In general, the daily oral dose of each active ingredient, when used for the indicated effect, will be between about 0.0001 to 1000mg/kg body weight, preferably between about 0.001 to 100mg/kg body weight per day, most preferably between about 0.1 to 20 mg/kg/day. For intravenous use, the most preferred dose will be from about 0.1 to about 10 mg/kg/minute, infused at a constant rate. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0, 10.0, 15.0.20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0 and 1000.0 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. The compounds may be administered on a regimen of 1 to 4 times per day, preferably once or twice per day.

The compounds of the present invention may also be administered intranasally, via topical use of a suitable intranasal vehicle, or via a transdermal route, e.g., using a transdermal skin patch. When administered in a transdermal delivery system, dosing can be continuous rather than intermittent throughout the dosage regimen.

The compounds of the present invention are generally administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as pharmaceutical carriers), suitably selected with respect to the intended mode of administration, i.e., oral tablets, capsules, elixirs, syrups, and the like, and consistent with conventional pharmaceutical practice.

For example, for oral administration in the form of tablets or capsules, the active pharmaceutical ingredient may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like. For oral administration in liquid form, the oral pharmaceutical composition may be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier, such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents may also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars (e.g., glucose or beta-lactose), corn sweeteners, natural and synthetic gums (e.g., acacia, tragacanth or sodium alginate), carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, without limitation, starch, methylcellulose, agar, bentonite, xanthan gum, and the like.

The compounds of the invention may also be provided to patients in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.

The compounds of the invention may also be coupled to soluble polymers suitable as targetable drug carriers. Such polymers may include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol, or polyoxyethylene-polylysine substituted with palmitoyl residues. In addition, the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, such as polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polycaprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans, and crosslinked or amphipathic block copolymers of hydrogels.

Dosage forms of the compounds of the invention suitable for administration may contain from about 0.1 mg to about 100mg of active ingredient per dosage unit. In such pharmaceutical compositions, the active ingredient will generally be present in an amount of about 0.5 to 95% by weight, based on the total weight of the composition.

Gelatin capsules may also be used as dosage forms which may contain the active ingredient and a milled carrier such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like. Compressed tablets may be prepared using similar diluents. Both tablets and capsules can be manufactured as sustained release products to provide continuous drug release over hours. Compressed tablets may be sugar-coated or film-coated to mask any unpleasant taste, to protect the tablet from the atmosphere, or enteric-coated for selective disintegration in the gastrointestinal tract.

When liquid dosage forms are used for oral administration, they may contain coloring and flavoring agents to increase patient acceptance.

Generally, water, suitable oils, saline, aqueous dextrose and related sugar solutions and glycols (e.g., propylene glycol or polyethylene glycol) are suitable carriers for parenteral solutions. The parenteral solutions preferably contain a water-soluble salt of the active ingredient, suitable stabilizers and, if necessary, buffer substances. Antioxidants are suitable stabilizers, such as sodium bisulfite, sodium sulfite, or ascorbic acid, alone or in combination. Citric acid and its salts and sodium EDTA are also used. In addition, parenteral solutions may contain preservatives such as benzalkonium chloride, methyl or propyl parabens, and chlorobutanol. Suitable pharmaceutical carriers are described in Remington's pharmaceutical Sciences, Mack Publishing Company, which is a standard reference in the pharmaceutical arts.

The pharmaceutical composition of the present invention may also be in the form of an oil-in-water emulsion. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin, or a mixture of these. Suitable emulsifying agents may be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally-occurring phosphatides, for example soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents.

The compounds of the invention may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.

For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the compounds of the present invention are employed. Topical application as used herein is also meant to include the use of mouth washes and gargles.

The pharmaceutical compositions and methods of the present invention may further comprise other therapeutically active compounds commonly used for the treatment of the above mentioned pathological conditions.

Representative pharmaceutical dosage forms useful for administration of the compounds of the present invention may be illustrated as follows:

capsule preparation

A large number of unit capsules can be prepared by filling each standard two-piece hard gelatin capsule with 50mg of the powdered active ingredient, 100mg of lactose, 25 mg of cellulose and 3mg of magnesium stearate.

Soft gelatin capsule

A mixture of the active ingredient in a digestible oil such as soybean oil, cottonseed oil or olive oil can be prepared and injected into the gelatin by means of a positive displacement pump to form soft gelatin capsules containing 75mg of the active ingredient. The capsules should be washed and dried.

Tablet formulation

Tablets may be prepared by conventional techniques so that the dosage unit is 75mg of active ingredient, 0.15 mg of colloidal silicon dioxide, 4mg of magnesium stearate, 250 mg of microcrystalline cellulose, 9mg of starch and 75mg of lactose. Suitable coatings well known to those skilled in the art may be applied to increase palatability or delay absorption.

Injection preparation

Parenteral compositions suitable for administration by injection may be prepared by stirring 1.0% by weight of the active ingredient in 8% by volume propylene glycol and water. The solution should be made isotonic with sodium chloride and sterilized.

Suspension liquid

An aqueous suspension may be prepared for oral administration so as to contain 75mg of finely divided active ingredient, 150mg of sodium carboxymethylcellulose, 3.75mg of sodium benzoate, 0.75g of sorbitol solution U.S.P. and 0.015mL of vanillin per 5 mL.

Example A

This example describes the evaluation of the efficacy of CCR2 antagonists for the treatment of rheumatoid arthritis.

An animal model of rheumatoid arthritis can be induced in rodents by injection of selected adjuvants containing type II collagen. Three series of rodents were injected subcutaneously or intradermally with type II collagen emulsified in complete freund's adjuvant on days 0 and 21, each group consisting of 15 genetically susceptible mice or rats. The first series of rodents additionally received Phosphate Buffered Saline (PBS) and tween 0.5% i.p. at initial sensitization, followed by a different dosing regimen. The second series consisted of rodent groups receiving different doses of CCR2 antagonist, administered intraperitoneally, intravenously, subcutaneously, intramuscularly, orally or via any other mode of administration at the time of initial sensitization, followed by a different dosing regimen. A third series of rodents served as positive controls, treated with mouse IL-10i.p. or anti-TNF antibody i.p. at initial sensitization, and then received a different dosing regimen.

Animals were monitored for 3 to 8 weeks of swollen joint or paw development, rated according to a standard disease severity scale. Disease severity was confirmed by histological analysis of the joints.

Another aspect of the invention relates to radiolabeled compounds of the invention which will find use not only in radioimaging, but also in assays, both in vitro and in vivo, for locating and quantifying chemokine receptors in tissue samples, including humans, and for identifying chemokine receptor ligands by inhibiting the binding of a radiolabeled compound. Accordingly, the invention includes chemokine receptor assays comprising such radiolabeled compounds.

The invention further includes isotopically-labeled compounds of the invention. An "isotopically" or "radio-labeled" compound is one in which one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature (i.e., naturally occurring). Suitable radionuclides that may be incorporated in the compounds of the present invention include, but are not limited to²H (also written as D or deuterium),³H (also written as T or tritium),¹¹C、¹³C、¹⁴C、¹³N、¹⁵N、¹⁵O、¹⁷O、¹⁸、¹⁸F、³⁵S、³⁶Cl、⁸²Br、⁷⁵Br、⁷⁶Br、⁷⁷Br、¹²³I、¹²⁴I、¹²⁵I and¹³¹I. the radionuclide that is incorporated into the radiolabeled compounds of the invention will depend on the particular application for which the radiolabeled compound is used. For example, for in vitro chemokine receptor labeling and competition assays, incorporation³H、¹⁴C、⁸²Br、¹²⁵I、¹³¹I、³⁵Compounds of S will generally be very useful. In the case of a radiological imaging application,¹¹C、¹⁸F、¹²⁵I、¹²³I、¹²⁴I、¹³¹I、⁷⁵Br、⁷⁶br or⁷⁷Br will generally be very useful。

It goes without saying that a "radiolabeled" or "labeled" compound is a compound that incorporates at least one radionuclide. In some embodiments, the radionuclide is selected from the group consisting of³H、¹⁴C、¹²⁵I、³⁵S and⁸²br.

Synthetic methods for incorporating radioisotopes into organic compounds are useful for the compounds of the present invention and are well known in the art.

The radiolabeled compounds of the invention may be used in screening assays to identify/evaluate compounds. In a general sense, a newly synthesized or identified compound (i.e., test compound) can be evaluated for its ability to reduce binding of a radiolabeled compound of the invention to a chemokine receptor. Thus, the ability of a test compound to compete with a radiolabeled compound for binding to a chemokine receptor is directly related to its binding affinity.

The invention also includes kits, e.g., useful for treating or preventing chemokine-associated diseases, which comprise one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I. Such kits may further include, if desired, one or more of various conventional kit components, such as a container containing one or more pharmaceutically acceptable carriers, additional containers, and the like, as will be apparent to those skilled in the art. Instructions may also be included in the kit, as an insert or as a label, indicating the amount of the components to be administered, directions for administration, and/or directions for mixing the components.

All publications, patents, and patent applications, including all technical and bibliographic documents cited herein, are hereby incorporated by reference in their entirety for all purposes.

While many of the inventive forms disclosed herein constitute presently preferred embodiments, many others are possible, and further details of the preferred and other possible embodiments are not to be construed as limiting. It is understood that the terms used herein are merely descriptive, rather than limiting, and that various equivalents may be substituted without departing from the spirit or scope of the claimed invention.

Claims (29)

1. A compound, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

n- (2- { (3S) -3- [ (4-hydroxy-4-phenylcyclohexyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) (methyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (methoxymethyl) pyridin-2-yl ] cyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4- {5- [ (dimethylamino) methyl ] pyridin-2-yl } -4-hydroxycyclohexyl) -amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (4-methylphenyl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4-hydroxy-4-pyridin-3-ylcyclohexyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-methylpyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (6-methylpyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (6-methoxypyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (6-methoxypyridin-3-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

3- (trifluoromethyl) -N- [2- ((3S) -3- { [ 4-hydroxy-4- (5-methyl-1, 3-thiazol-2-yl) cyclohexyl ] -amino } pyrrolidin-1-yl) -2-oxoethyl ] benzamide,

3- (trifluoromethyl) -N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1-hydroxy-1-methylethyl) -1, 3-thiazol-2-yl ] cyclohexyl } amino) pyrrolidin-1-yl ] -2-oxoethyl } benzamide,

3- (trifluoromethyl) -N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (methoxymethyl) -1, 3-thiazol-2-yl ] cyclohexyl } amino) pyrrolidin-1-yl ] -2-oxoethyl } benzamide,

n-ethyl-2- (1-hydroxy-4- { [ (3S) -1- ({ [3- (trifluoromethyl) benzoyl ] amino } acetyl) pyrrolidin-3-yl ] amino } cyclohexyl) -1, 3-thiazole-5-amide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (pyrrolidin-1-ylcarbonyl) -1, 3-thiazol-2-yl ] cyclohexyl } -amino) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- [2- (3S) - (3- { [ 4-hydroxy-4- (5-pyridin-3-yl-1, 3-thiazol-2-yl) cyclohexyl ] methyl } -pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ({ (3S) -1- [ 4-hydroxy-4- (5-pyridin-2-yl-1, 3-thiazol-2-yl) cyclohexyl ] pyrrolidin-3-yl } amino) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4-hydroxy-4-pyrazin-2-cyclohexylamino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4-hydroxy-4-pyrimidin-2-ylcyclohexyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

6- (1-hydroxy-4- { [ (3S) -1- ({ [3- (trifluoromethyl) benzoyl ] amino } acetyl) pyrrolidin-3-yl ] amino } cyclohexyl) -N, N-dimethylnicotinamide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (pyrrolidin-1-ylcarbonyl) pyridin-2-yl ] cyclohexyl } -amino) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- [4- (5-bromopyridin-2-yl) -4-hydroxycyclohexyl ] aminopyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({4- [5- (2-formylphenyl) pyridin-2-yl ] -4-hydroxycyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- (3S) -3- [ (4-hydroxy-4-5- [2- (hydroxymethyl) phenyl ] pyridin-2-cyclohexyl) -amino ] pyrrolidin-1-yl-2-oxoethyl) -3- (trifluoromethyl) benzamide bis (trifluoroacetate),

n- [2- ((3S) -3- [ 4-hydroxy-4- (4-pyrimidin-2-ylphenyl) cyclohexyl ] aminopyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide bis (trifluoroacetate),

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1, 3-thiazol-2-yl) pyridin-2-yl ] cyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } -pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4- {5- [3- (aminocarbonyl) phenyl ] pyridin-2-yl } -4-hydroxycyclohexyl) -amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4- {5- [2- (aminocarbonyl) phenyl ] pyridin-2-yl } -4-hydroxycyclohexyl) -amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({4- [5- (3-acetylphenyl) pyridin-2-yl ] -4-hydroxycyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

3- [6- (1-hydroxy-4- { [ (3S) -1- ({ [3- (trifluoromethyl) benzoyl ] amino } acetyl) pyrrolidin-3-yl ] amino } cyclohexyl) pyridin-3-yl ] benzoic acid,

n- (2- { (3S) -3- [ (4-hydroxy-4- {5- [3- (hydroxymethyl) phenyl ] pyridin-2-yl } cyclohexyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrimidin-5-ylpyridin-2-yl) cyclohexyl ] amino } -pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [4- (3,3' -bipyridin-6-yl) -4-hydroxycyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [4- (3,4' -bipyridin-6-yl) -4-hydroxycyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrazin-2-ylpyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [4- (1H-imidazol-1-yl) phenyl ] cyclohexyl } amino) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [4- (2' -formylbiphenyl-4-yl) -4-hydroxycyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [2' - (hydroxymethyl) biphenyl-4-yl ] cyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({4- [5- (3, 5-Dimethyliso-yl)Azol-4-yl) pyridin-2-yl]-4-hydroxycyclohexyl } -amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1, 3-)Azol-2-yl) pyridin-2-yl]Cyclohexyl } amino) -pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (2R,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2-methylpyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2- (methoxymethyl) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -2- (ethoxymethyl) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -2- (1-hydroxy-1-methylethyl) -4- [ (trans-4-hydroxy-4-pyridin-2-ylcyclohexyl) -amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -2- [ 1-hydroxyethyl ] -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2- (1-methoxy-1-methyl-ethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2- [ (1S) -1-methoxyethyl ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2-isopropylpyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- ({ 4-hydroxy-4- [5- (methoxymethyl) pyridin-2-yl ] cyclohexyl } amino) -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- [ (4- {5- [ (dimethylamino) methyl ] pyridin-2-yl } -4-hydroxycyclohexyl) -amino ] -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2- (isopropoxymethyl) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ [ 4-hydroxy-4- (5-pyrazin-2-ylpyridin-2-yl) cyclohexyl ] (methyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ { 4-hydroxy-4- [5- (1, 3-)Azol-2-yl) pyridin-2-yl]Cyclohexyl } (methyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1-[5-(1,3-azol-2-yl) pyridin-2-yl]-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl]Piperidin-4-yl } carbonyl) pyrrolidin-3-yl]Amino group(s) of a cyclohexanol alcohol,

1- (5-pyrazin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-methylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (3,3' -bipyridin-6-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (3,4' -bipyridin-6-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-methoxypyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- [5- (methoxymethyl) pyridin-2-yl ] -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

6- (1-hydroxy-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexyl) nicotinamide,

6- (1-hydroxy-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexyl) -N-methylnicotinamide,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyrimidin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [5- (trifluoromethyl) pyridin-2-yl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyrimidin-2-yl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] azetidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol, and

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({ [4- (trifluoromethyl) pyridin-2-yl ] methoxy } acetyl) pyrrolidin-3-yl ] amino } cyclohexanol.

2. A compound, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

n- (2- { (3S) -3- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) (methyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (methoxymethyl) pyridin-2-yl ] cyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4- {5- [ (dimethylamino) methyl ] pyridin-2-yl } -4-hydroxycyclohexyl) -amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-methylpyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (6-methoxypyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

3- (trifluoromethyl) -N- [2- ((3S) -3- { [ 4-hydroxy-4- (5-methyl-1, 3-thiazol-2-yl) cyclohexyl ] -amino } pyrrolidin-1-yl) -2-oxoethyl ] benzamide,

3- (trifluoromethyl) -N- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (methoxymethyl) -1, 3-thiazol-2-yl ] cyclohexyl } amino) pyrrolidin-1-yl ] -2-oxoethyl } benzamide,

n- [2- (3S) - (3- { [ 4-hydroxy-4- (5-pyridin-3-yl-1, 3-thiazol-2-yl) cyclohexyl ] methyl } -pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ({ (3S) -1- [ 4-hydroxy-4- (5-pyridin-2-yl-1, 3-thiazol-2-yl) cyclohexyl ] pyrrolidin-3-yl } amino) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

6- (1-hydroxy-4- { [ (3S) -1- ({ [3- (trifluoromethyl) benzoyl ] amino } acetyl) pyrrolidin-3-yl ] amino } cyclohexyl) -N, N-dimethylnicotinamide,

n- [2- ((3S) -3- [4- (5-bromopyridin-2-yl) -4-hydroxycyclohexyl ] aminopyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({4- [5- (2-formylphenyl) pyridin-2-yl ] -4-hydroxycyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- (3S) -3- [ (4-hydroxy-4-5- [2- (hydroxymethyl) phenyl ] pyridin-2-cyclohexyl) -amino ] pyrrolidin-1-yl-2-oxoethyl) -3- (trifluoromethyl) benzamide bis (trifluoroacetate),

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1, 3-thiazol-2-yl) pyridin-2-yl ] cyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } -pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4- {5- [3- (aminocarbonyl) phenyl ] pyridin-2-yl } -4-hydroxycyclohexyl) -amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({4- [5- (3-acetylphenyl) pyridin-2-yl ] -4-hydroxycyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4-hydroxy-4- {5- [3- (hydroxymethyl) phenyl ] pyridin-2-yl } cyclohexyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrimidin-5-ylpyridin-2-yl) cyclohexyl ] amino } -pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [4- (3,3' -bipyridin-6-yl) -4-hydroxycyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [4- (3,4' -bipyridin-6-yl) -4-hydroxycyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrazin-2-ylpyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [4- (1H-imidazol-1-yl) phenyl ] cyclohexyl } amino) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({4- [5- (3, 5-Dimethyliso-yl)Azol-4-yl) pyridin-2-yl]-4-hydroxycyclohexyl } -amino) pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1, 3-)Azol-2-yl) pyridin-2-yl]Cyclohexyl } amino) -pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (2R,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2-methylpyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2- (methoxymethyl) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -2- (ethoxymethyl) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -2- [ 1-hydroxyethyl ] -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2- [ (1S) -1-methoxyethyl ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2-isopropylpyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- ({ 4-hydroxy-4- [5- (methoxymethyl) pyridin-2-yl ] cyclohexyl } amino) -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- [ (4- {5- [ (dimethylamino) methyl ] pyridin-2-yl } -4-hydroxycyclohexyl) -amino ] -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ [ 4-hydroxy-4- (5-pyrazin-2-ylpyridin-2-yl) cyclohexyl ] (methyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ { 4-hydroxy-4- [5- (1, 3-)Azol-2-yl) pyridin-2-yl]Cyclohexyl } (methyl) amino]Pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1-[5-(1,3-azol-2-yl) pyridin-2-yl]-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl]Piperidin-4-yl } carbonyl) pyrrolidin-3-yl]Amino group(s) of a cyclohexanol alcohol,

1- (5-pyrazin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-methylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (3,3' -bipyridin-6-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (3,4' -bipyridin-6-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-methoxypyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- [5- (methoxymethyl) pyridin-2-yl ] -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

6- (1-hydroxy-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexyl) -N-methylnicotinamide,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyrimidin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyrimidin-2-yl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol, and

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] azetidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol.

3. A compound, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (methoxymethyl) pyridin-2-yl ] cyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-methylpyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- (2- (3S) -3- [ (4-hydroxy-4-5- [2- (hydroxymethyl) phenyl ] pyridin-2-cyclohexyl) -amino ] pyrrolidin-1-yl-2-oxoethyl) -3- (trifluoromethyl) benzamide bis (trifluoroacetate),

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1, 3-thiazol-2-yl) pyridin-2-yl ] cyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } -pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({4- [5- (3-acetylphenyl) pyridin-2-yl ] -4-hydroxycyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ (4-hydroxy-4- {5- [3- (hydroxymethyl) phenyl ] pyridin-2-yl } cyclohexyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrimidin-5-ylpyridin-2-yl) cyclohexyl ] amino } -pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [4- (3,4' -bipyridin-6-yl) -4-hydroxycyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrazin-2-ylpyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1, 3-)Azol-2-yl) pyridin-2-yl]Cyclohexyl } amino) -pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -2- (ethoxymethyl) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -2- [ 1-hydroxyethyl ] -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2- [ (1S) -1-methoxyethyl ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- ({ 4-hydroxy-4- [5- (methoxymethyl) pyridin-2-yl ] cyclohexyl } amino) -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- [ (4- {5- [ (dimethylamino) methyl ] pyridin-2-yl } -4-hydroxycyclohexyl) -amino ] -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (3S) -3- [ [ 4-hydroxy-4- (5-pyrazin-2-ylpyridin-2-yl) cyclohexyl ] (methyl) amino ] pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1-pyridin-2-yl-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1-[5-(1,3-azol-2-yl) pyridin-2-yl]-4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl]Piperidin-4-yl } carbonyl) pyrrolidin-3-yl]Amino group(s) of a cyclohexanol alcohol,

1- (5-pyrazin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-methylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (3,3' -bipyridin-6-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (3,4' -bipyridin-6-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-methoxypyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- [5- (methoxymethyl) pyridin-2-yl ] -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol, and

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyrimidin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol.

4. A compound, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } -pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({4- [5- (3-acetylphenyl) pyridin-2-yl ] -4-hydroxycyclohexyl } amino) -pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [4- (3,4' -bipyridin-6-yl) -4-hydroxycyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrazin-2-ylpyridin-2-yl) cyclohexyl ] amino } pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide,

n- {2- [ (3S) -3- ({ 4-hydroxy-4- [5- (1, 3-)Azol-2-yl) pyridin-2-yl]Cyclohexyl } amino) -pyrrolidin-1-yl]-2-oxoethyl } -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -2- (ethoxymethyl) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -2- [ 1-hydroxyethyl ] -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- (2- { (2S,4S) -4- [ (4-hydroxy-4-pyridin-2-ylcyclohexyl) amino ] -2- [ (1S) -1-methoxyethyl ] -pyrrolidin-1-yl } -2-oxoethyl) -3- (trifluoromethyl) benzamide,

n- {2- [ (2S,4S) -4- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } -2- (methoxymethyl) pyrrolidin-1-yl ] -2-oxoethyl } -3- (trifluoromethyl) benzamide,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [3- (trifluoromethyl) phenyl ] pyrrolidin-3-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [6- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-pyrazin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-methylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (3,3' -bipyridin-6-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- (5-methoxypyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol,

1- [5- (methoxymethyl) pyridin-2-yl ] -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyridin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol, and

1- (5-pyrimidin-2-ylpyridin-2-yl) -4- { [ (3S) -1- ({1- [4- (trifluoromethyl) pyrimidin-2-yl ] piperidin-4-yl } carbonyl) pyrrolidin-3-yl ] amino } cyclohexanol.

N- [2- ((3S) -3- { [ 4-hydroxy-4- (5-pyrimidin-2-ylpyridin-2-yl) cyclohexyl ] amino } -pyrrolidin-1-yl) -2-oxoethyl ] -3- (trifluoromethyl) benzamide, or a pharmaceutically acceptable salt thereof.

6. A composition comprising a compound of claim 2 and a pharmaceutically acceptable carrier.

7. A composition comprising a compound of claim 3 and a pharmaceutically acceptable carrier.

8. A composition comprising a compound of claim 4 and a pharmaceutically acceptable carrier.

9. A composition comprising a compound of claim 5 and a pharmaceutically acceptable carrier.

10. Use of a compound of claim 1 in the manufacture of a medicament for treating inflammation, atherosclerosis, neuropathic pain, lupus, restenosis, immune disorders, and graft rejection in a mammal in need thereof.

11. Use of a compound of claim 1 in the manufacture of a medicament for treating rheumatoid arthritis or systemic lupus erythematosus in a mammal in need thereof.

12. Use of a compound of claim 1 in the manufacture of a medicament for modulating chemokine receptor activity in a mammal.

13. Use of a compound of claim 1 in the manufacture of a medicament for treating a CCR2 mediated condition or disease in a subject.

14. Use of a compound of claim 1 in the manufacture of a medicament for treating a CCR5 mediated condition or disease in a subject.

15. Use of a compound according to claim 1 in the manufacture of a medicament for modulating chemokine receptor activity.

16. The use of claim 15 wherein the chemokine receptor is CCR2 or CCR 5.

17. The use of claim 15, wherein said modulation is inhibition.

18. Use of a compound according to claim 1 in the manufacture of a medicament for the treatment of a disease associated with expression or activity of a chemokine receptor in a patient.

19. The use of claim 18 wherein the chemokine receptor is CCR2 or CCR 5.

20. The use of claim 18, wherein the disease is an inflammatory disease.

21. The use of claim 18, wherein the disease is an immune disorder.

22. The use of claim 18, wherein the disease is rheumatoid arthritis, atherosclerosis, lupus, multiple sclerosis, neuropathic pain, graft rejection, diabetes or obesity.

23. The use of claim 18, wherein the disease is cancer.

24. The use of claim 23, wherein the cancer is characterized by tumor-associated macrophages.

25. The use of claim 23, wherein the cancer is breast cancer, ovarian cancer, or multiple myeloma.

26. The use of claim 18, wherein the disease or condition is a viral infection.

27. The use of claim 26, wherein the viral infection is an HIV infection.

28. Use of a compound according to claim 1 in the manufacture of a medicament for the treatment of HIV infection in a patient.

29. The use of claim 28, wherein the treatment further comprises the simultaneous or sequential administration of at least one antiviral agent.