CN1976702A

CN1976702A - 3-aminocyclopentanecarboxamides as modulators of chemotactic factor receptors

Info

Publication number: CN1976702A
Application number: CN 200580021744
Authority: CN
Inventors: 薛楚标; 郑长胜; 冯皓; M·夏; J·戈兰恩; 曹敢峰; B·W·迈特卡夫
Original assignee: Insight Inc
Current assignee: Incyte Corp; Insight Inc
Priority date: 2004-06-28
Filing date: 2005-06-27
Publication date: 2007-06-06
Also published as: CN1976707A; TNSN06444A1; UA83142C2; ZA200610667B; TNSN06443A1; ZA200610426B

Abstract

The present invention is directed to compounds of formula (I): which are modulators of chemokine receptors. The compounds of the invention, and compositions thereof, are useful in the treatment of diseases related to chemokine receptor expression and/or activity.

Description

3-aminocyclopentanecarasxamides as chemokine receptor modulators

Background of invention

The present invention relates to regulate chemokine receptors, such as the active chemical compound of CCR2 and CCR5.In obvious embodiment, these chemical compounds are regulated CCR2 and CCR5.For example, these chemical compounds are used for the treatment of and chemokine receptor expression or active relevant disease.

Background of invention

Leukocyte moves and transports illing tissue from blood vessel and relates to the normal disease-resistant inflammatory reaction of startup.This process is also referred to as the leukocyte recruitment reaction, and it also relates to outbreak and the development and the weak systemic autoimmune disease of life-threatening inflammation.The pathologic condition that these diseases produce derives from the attack of health immune system to normal structure.Therefore, prevent that the target tissue of raising to inflammation, autoimmune disease and the cancer with blocking leukocyte from can become the effective means of height that treatment gets involved.

The dissimilar leukocyte that relates to cell immune response comprises mononuclear cell, lymphocyte, neutrophilic granulocyte, oxyphil cell and basophil.In most of situation, mononuclear cell and lymphocyte be for starting, coordinate and keep the leukocyte type of chronic inflammatory reaction, and blocking these cells, to enter inflammation part be ideal.Lymphocyte attracts mononuclear cell to reach tissue site, and they and lymphocyte cause occurring in the most of actual tissue infringement in the inflammatory diseases jointly.Known lymphocyte and/or monocyte infiltration can cause chronic widely autoimmune disease and can also cause the organ-graft refection.These diseases include but not limited to rheumatoid arthritis, chronic contact dermatitis, inflammatory bowel, lupus, systemic lupus erythematosus (sle), multiple sclerosis, atherosclerosis, psoriasis, sarcoidosis, idiopathic pulmonary fibrosis, dermatomyositis, skin pemphigoid and relevant disease (pemphigus vulgaris (Pemphigus vulgaris) for example, pemphigus foliaceus (P.foliacious), pemphigus erythematosus (P.erythematosis)), glomerulonephritis, vasculitis, hepatitis, diabetes, allograft rejection and graft versus host disease.

Think that the process that leukocyte leaves blood flow, is accumulated in inflammation part and starts disease had at least three steps, they are described as: roll (1); (2) activation/firmly adhesion; (3) stride endothelial migration [Springer, T.A., Nature 346:425-433 (1990); Lawrence and Springer, Cell 65:859-873 (1991); Butcher, E.C., Cell67:1033-1036 (1991)].Second step is receptor-mediated by chemoattractant on molecular level.Chemoattractant receptor on the leukocyte surface is subsequently in conjunction with the chemoattracting cytoking by the emiocytosis on infringement or the infection site.The activated leukocyte cell of receptors bind increases the cohesiveness that the adhesion molecule of endothelial migration is striden in mediation, and promotes that cell directional migrates to the chemoattracting cytoking source.

Chemoattracting cytoking (leukocyte chemoattractant/activation factor) is also referred to as chemotactic factor, be also referred to as mutual secretin (intercrine) and SIS cytokine, they are that one group of molecular weight is inflammation/immunoloregulation polypeptide factor of 6-15kDa, discharge by the various cells on the inflammation part, such as macrophage, mononuclear cell, the oxyphil cell, neutrophilic granulocyte, fibroblast, vascular endothelial cell, epithelial cell, (summary is at Luster for smooth muscle cell and mastocyte, New Eng.J Med., 338,436-445 (1998) and Rollins, Blood, 90, among the 909-928 (1997)).In addition, chemotactic factor is described in the following document: Oppenheim, J.J. etc., Annu.Rev.Immunol., 9:617-648 (1991); Schall and Bacon, Curr.Opin.Immunol., 6:865-873 (1994); Baggiolini, M. etc., and Adv.Immunol., 55:97-179 (1994).Chemotactic factor has the migration of the committed cell of stimulation, is also referred to as the ability of chemotactic process.Every kind of cytokine contains 4 cysteine residues (C) and 2 inner disulfide bond.Can be based on two amino terminal cysteine residues whether closely adjacent (CC family) or separated (CXC family) by an aminoacid chemotactic factor is divided into two groups of main subtribes.It is relevant that these differences and two subtribes constitute independent gene cluster.In each gene cluster, chemotactic factor generally shows the sequence similarity of 25-60%.The CXC chemotactic factor, mainly neutrophilic leukocyte and T lymphocyte had chemotaxis such as interleukin 8 (IL-8), neutrophilic leukocyte-activated protein-2 (NAP-2) and melanoma growth-stimulating activity albumen (MGSA), and the CC chemotactic factor, such as RANTES, MIP-1 α, MIP-1 β, monocyte chemoattractant protein (MCP-1, MCP-2, MCP-3, MCP-4, and MCP-5) and eotaxin (1 and-2) cell types such as macrophage, T lymphocyte, oxyphil cell, dendritic cell and basophil had chemotaxis.Also there is the chemotactic factor that does not belong to main chemotactic factor subtribe: lymphocyte chemotactic factor (LCF)-1, lymphocyte chemotactic factor (LCF)-2 (being the C chemotactic factor); With fractalkine chemotactic molecule (CXXXC chemotactic factor).

MCP-1 (is also referred to as MCAF (abbreviation of macrophage chemotactic and activation factor) or the CC chemotactic factor that JE) produces for monocyte/macrophage, smooth muscle cell, fibroblast and vascular endothelial cell and causes mononuclear cell (for example, referring to Valente, A.J. etc., Biochemistry, 1988,27,4162; Matsushima, K. etc., J.Exp.Med., 1989,169,1485; Yoshimura, T. etc., J.Immunol., 1989,142,1956; Rollins, B.J. etc., Proc.Natl.Acad.Sci.USA, 1988,85,3738; Rollins, B.J. etc., Blood, 1991,78,1112; Jiang, Y. etc., J.Immunol., 1992,148,2423; Vaddi, K. etc., J.Immunol., 1994,153,4721), the memory T lymphocyte (for example, referring to Carr, M.W. etc., Proc.Natl.Acad.Sci.USA, 1994,91,3652), the T lymphocyte (for example, referring to Loetscher, P. etc., FASEB J., 1994,8,1055) and natural killer cell (for example, referring to Loetscher, P. etc., J.Immunol., 1996,156,322; Allavena, P. etc., Eur.J.Immunol., 1994,24,3233) cell migration and cell adhesion, and the histamine release of mediation basophil (for example, referring to Alam, R. etc., J.Clin.Invest., 1992,89,723; Bischoff, S.C. etc., J.Exp.Med., 1992,175,1271; Kuna, P. etc., J.Exp.Med., 1992,175,489).In addition, having reported the height of MCP-1 in disease expresses, think that wherein monocyte/macrophage and/or T cellular accumulation are important in disease takes place or develops, such as atherosclerosis (for example, referring to Hayes, I.M. etc., Arterioscler.Thromb.Vasc.Biol., 1998,18,397; Takeya, M.. etc., Hum.Pathol., 1993,24,534; Yla-Herttuala, S. etc., Proc.Natl.Acad.Sci.USA, 1991,88,5252; Nelken, N.A., J.Clin.Invest., 1991,88,1121), rheumatoid arthritis (for example, referring to Koch, A.E. etc., J.Clin.Invest., 1992,90,772; Akahoshi, T. etc., Arthritis Rheum., 1993,36,762; Robinson, E. etc., Clin.Exp.Immunol., 101,398), nephritis (for example, referring to Noris, M. etc., Lab.Invest., 1995,73,804; Wada, T. etc., Kidney Int., 1996,49,761; Gesualdo, L. etc., Kidney Int., 1997,51,155), nephropathy (for example, referring to Saitoh, A. etc., J.Clin.Lab.Anal., 1998,12,1; Yokoyama, H. etc., J.Leukoc.Biol., 1998,63,493), pulmonary fibrosis, pulmonary sarcoidosis are (for example, referring to Sugiyama, Y. etc., Internal Medicine, 1997,36,856), asthma (for example, referring to Karina, M. etc., J.Invest.Allergol.Clin.Immunol., 1997,7,254; Stephene, T.H., Am.J.Respir.Crit.Care Med., 1997,156,1377; Sousa, A.R. etc., Am.J.Respir.CellMol.Biol., 1994,10,142), multiple sclerosis (for example, referring to McManus, C. etc., J.Neuroimmunol., 1998,86,20), psoriasis (for example, referring to Gillitzer, R. etc., J.Invest.Dermatol., 1993,101,127), inflammatory bowel (for example, referring to Grimm, M.C. etc., J.Leukoc.Biol., 1996,59,804; Reinecker, H.C. etc., Gastroenterology, 1995,106,40), myocarditis (for example, referring to Seino, Y. etc., Cytokine, 1995,7,301), endometriosis (for example, referring to Jolicoeur, C. etc., Am.J.Pathol., 1998,152,125), the intraperitoneal adhesion (for example, referring to Zeyneloglu, H.B. etc., Human Reproduction, 1998,13,1194), congestive heart failure (for example, referring to Aurust, P. etc., Circulation, 1998,97,1136), chronic hepatopathy (for example, referring to Marra, F. etc., Am.J.Pathol., 1998,152,423), viral meningitis (for example, referring to Lahrtz, F. etc., Eur.J.Immunol., 1997,27,2484), kawasaki disease (for example, referring to Wong, M.; Deng, J.Rheumatol., 1997,24,1179) and sepsis (for example, referring to Salkowski, C.A.; Deng, Infect.Immun., 1998,66,3569).In addition, reported anti--MCP-1 antibody and in the animal model of following disease, shown inhibitory action or therapeutical effect: rheumatoid arthritis (for example, referring to Schimmer, R.C. etc., J.Immunol., 1998,160,1466; Schrier, D.J., J.Leukoc.Biol., 1998,63,359; Ogata, H. etc., J.Pathol., 1997,182,106); Multiple sclerosis (for example, referring to Karpus, W.J. etc., J.Leukoc.Biol., 1997,62,681); Nephritis (for example, referring to Lloyd, C.M. etc., J.Exp.Med., 1997,185,1371; Wada, T. etc., FASEB J., 1996,10,1418); Asthma (for example, referring to Gonzalo, J.-A. etc., J.Exp.Med., 1998,188,157; Lukacs, N.W., J.Immunol., 1997,158,4398); Atherosclerosis (for example, referring to Guzman, L.A. etc., Circulation, 1993,88 (suppl.), I-371); Delayed hypersensitivity (for example, referring to Rand, M.L. etc., Am.J.Pathol., 1996,148,855); Pulmonary hypertension (for example, referring to Kimura, H. etc., Lab.Invest., 1998,78,571); With intraperitoneal adhesion (for example, referring to Zeyneloglu, H.B. etc., Am.J.Obstet.Gynecol., 1998,179,438).The peptide antagonists MCP-1 (9-76) that has also reported MCP-1 in mouse model, suppress arthritis (referring to Gong, J.-H., J.Exp., 4ed., 1997,186,131), and the research in the MCP-1-deficient mice has confirmed that MCP-1 is mainly used in the body monocyte recruitement (referring to Lu, B. etc., J.Exp.Med., 1998,187,601; Gu, L. etc., Moll.Cell, 1998,2,275).

Chronic obstructive pulmonary disease (COPD) is arranged as modal grade in the cause of death of Western society.It is defined as the decline of carrying out property of pulmonary function, and only part can reverse by bronchodilator.COPD is characterised in that the chronic inflammatory disease that is different from asthma observed air flue or the alveolar, comprises that neutrophilic granulocyte, macrophage, CD8+T cell and/or the mastocyte quantity in airway walls, alveolar chamber and the vascular smooth muscle increases.Think that the cytokine relevant with COPD comprises tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-1 β, IL-6, IL-8 and MCP-1.Known CCR2 be the receptor of MCP-1 and in the recent period data supported MCP-1 and CCR2 directly or by the effect of macrophage in Airway Remodeling and inflammation.Therefore, the CCR2 antagonist for the treatment COPD attractive means (De Boer, W.I., Chest, 2002,121,209S-218S).

Document indication chemotactic factor, attract mononuclear cell and lymphocyte to reach disease location and mediate its activation such as MCP-1 and MIP-1 α, and think thus and closely relate to mononuclear cell and lymphocytic disease and take place, develop and keep and be closely related, such as atherosclerosis, diabetes, restenosis, rheumatoid arthritis, psoriasis, asthma, ulcerative colitis, nephritis (nephropathy), multiple sclerosis, pulmonary fibrosis, myocarditis, hepatitis, pancreatitis, sarcoidosis, Crohn disease, endometriosis, congestive heart failure, viral meningitis, cerebral infarction, neuropathy, kawasaki disease and sepsis are (for example, referring to Rovin, B.H. etc., Am.J.Kidney.Dis., 1998,31,1065; Lloyd, C. etc., Curr.Opin.Nephrol.Hypertens., 1998,7,281; Conti, P. etc., Allergy and Asthma Proc., 1998,19,121; Ransohoff, R.M. etc., Trends Neurosci., 1998,21,154; MacDermott, R.P. etc., Inflammatory Bowel Diseases, 1998,4,54).

Chemotactic factor is in conjunction with the specific cell surface receptor that belongs to G-albumen-link coupled seven-stride film district albumen family (summary is at Horuk, and Trends Pharm.Sci. is 15, among the 159-165 (1994)), and they are called " chemokine receptors ".In conjunction with its cognate ligand the time, chemokine receptors is by signal in the relevant trimerizing G protein transduction cell, thereby causes cellular calcium concentration to increase fast, and cell shape changes, and expression of cell adhesion molecules increases, threshing and promote reaction such as cell migration.

Cloned the gene of coding specificity chemokine receptors, and known these receptors are the G albumen-link coupled seven-transmembrane receptor that is present on the various leukocyte population.Up to now, at least 5 kinds of CXC chemokine receptors (CXCR1-CXCR5) and 8 kinds of CC-chemokine receptor (CCR1-CCR10) have been identified.For example, IL-8 is the part of CXCR1 and CXCR2, and MIP-1 α is the part of CCR1 and CCR5, and MCP-1 be CCR2A and CCR2B part (with regard to list of references, for example, referring to Holmes, W.E. etc., Science 1991,253,1278-1280; Murphy P.M. etc., Science, 253,1280-1283; Neote, K. etc., Cell, 1993,72,415-425; Charo, I.F. etc., Proc.Natl.Acad.Sci.USA, 1994,91,2752-2756; Yamagami, S. etc., Biochem.Biophys.Res.Commun., 1994,202,1156-1162; Combadier, C. etc., The Journal ofBiological Chemistry, 1995,270,16491-16494, Power, C.A. etc., J.Biol.Chem., 1995,270,19495-19500; Samson, M. etc., Biochemistry, 1996,35,3362-3367; Murphy, P.M., Annual Reviewof Immunology, 1994,12,592-633).According to reports lung inflammation and granuloma (granuroma) be formed in the CCR1-deficient mice and be inhibited (referring to Gao, J.-L. etc., J.Exp.Med., 1997,185,1959; Gerard, C. etc., J.Clin.Invest., 1997,100,2022), and macrophage raise and be formed in the CCR2-deficient mice with atherosclerotic lesion and reduce (referring to Boring, L. etc., Nature, 1998,394,894; Kuziel, W.A. etc., Proc.Natl.Acad.Sci., USA, 1997,94,12053; Kurihara, T. etc., J.Exp.Med., 1997,186,1757; Boring, L. etc., J.Clin.Invest., 1997,100,2552).

Chemokine receptors is also referred to as the coreceptor that the virus that causes viral infection, for example HIV to infect enters.Reverse transcription and protein processing are that the agent of design antiretroviral therapy is used for the classical step to its viral biocycle of blocking.Although think that many new drugs have the blocking virus of hope and enter, also there is not HIV-1 can't obtain the activating agent of resistance at present to it.Need a plurality of virus replication circulations could produce the hereditary difference that forms this resistance basis.Duplicate the conjoint therapy that is subjected to suppressing has to greatest extent kept using entry inhibitor when using other activating agent foundation stone.Think that a plurality of steps that targeting virus enters in the process have synergistic potential (Starr-Spires etc., Clin.Lab.Med., 2002,22 (3), 681).

HIV-1 enter CD4 (+) cell need viral envelope glycoprotein and CD4 and coreceptor, such as the interaction successively of chemokine receptor CCR 5 and CXCR4.The reasonable means of blocking this process are to use the micromolecule antagonist of coreceptor function.The TAK-779 molecule has been a kind of this class antagonist of the CCR5 of the effect infected of prevention HIV-1.TAK-779 duplicates by the interaction inhibition HIV-1 of blocking virus surface glycoprotein gp120 and CCR5.The binding site of the last TAK-779 of CCR5 is near the intracavity this receptor born of the same parents outer surface that forms between transbilayer helix 1,2,3 and 7 (Dragic etc., Proc.Natl.Acad.Sci.USA, 2000,97 (10), 5639).

Think that chemokine receptors CXCR4 and CCR5 are used separately as the T cell-tropism (X4) that enters its host cell and the coreceptor of macrophage-tropism (R5) HIV-1 bacterial strain.The bacterial strain of HIV-1 is bred on CD4 lymphocyte and macrophage needs the CCR5 coreceptor to express on cell surface.The individuality (CCR5 δ 32 homozygous genotypes) that lacks CCR5 is that phenotype is normal and the anti-HIV-1 infection.Virus enters the inhibition of the native ligand that can be subjected to CXCR4 (CXC chemotactic factor SDF-1) and CCR5 (CC chemotactic factor RANTES, MIP-1 α and MIP-1 β).With CXCR4 interactional first kind of non-peptide compound not taking place with CCR5 is quaternary ammonium derivative, is called TAK-779, and it also has effect and variable anti-HIV activity (De Clercq etc., Antivir.Chem.Chemother.2001,12 Suppl.1,19.

The micromolecular inhibitor that SCH-C (SCH 351125) enters for the another kind of HIV-1 by the CCR5 coreceptor.A kind of oxime-piperidine compounds SCH-C is the specific C CR5 antagonist of determining as in polyceptor combination and signal transduction test.The HIV-1 that this compound specificity suppresses CCR5 mediation in the U-87 astrocytic glioma infects, but the cell infection of expressing CXCR4 is not acted on (Strizki etc., Proc.Natl.Acad.Sci.USA, 2001,98 (22), 12718 or Tremblay etc., Antimicrobial Agents and Chemotherapy, 2002,46 (5), 1336).

With SCH-C at chemically relevant AD101 also entering by people CCR5 HIV inhibiting 1 type (HIV-1).Have been found that AD101 suppresses HIV-1 by macaque CCR5 and enters, and SCH-C can not.In 8 residues that there are differences between people and macaque coreceptor form, only have one, promptly methionine-198 can cause macaque CCR5 insensitive to the inhibitory action of SCH-C.198 stride film (TM) spiral 5 and are not arranged in the AD101 of the residue that relates to TM spiral 1,2,3 and 7 and above-mentioned definite binding site of SCH-C for being arranged in CCR5.Can influence the conformational state (Billick etc., 2004, J.Virol., 78 (8), 4134) of this receptor near the CCR5 district of 198 residues based on the research of the aminoacid replacement among CCR5 prompting.

The ideal medicament design means of the pharmaceutically active agents research and development requirement that is used for the treatment of the disease relevant with chemokine receptor activity has been represented in the evaluation of regulating the chemical compound of chemokine receptor activity.Chemical compound of the present invention helps to satisfy these and other requirement.

Summary of the invention

The invention provides the chemical compound of general formula I:

Or its pharmaceutically acceptable salt or prodrug, wherein this paper provides the formation member.

The present invention further provides the compositions that comprises compound of Formula I and pharmaceutically acceptable carrier.

The present invention further provides the method for regulating chemokine receptor activity, comprised the chemical compound that makes described chemokine receptors contact general formula I.

The present invention further provides the method for treatment and patient's chemokine receptor expression or active relevant disease, comprised the chemical compound of the general formula I that gives this patient treatment effective dose.

The present invention further provides the method that treatment patient HIV infects, comprised giving the chemical compound that this patient treats the general formula I of effective dose.

The present invention further provides the application of chemical compound as herein described in therapy.

The present invention further provides chemical compound as herein described is used for the medicine of therapy in preparation application.

Describe in detail

Chemical compound

The present invention provides the chemical compound of general formula I especially:

Or its pharmaceutically acceptable salt or prodrug, wherein:

Dotted line is represented the key chosen wantonly;

W is:

V is N, NO or CR ⁵

X is N, NO or CR ²

Y is N, NO or CR ³

Z is N, NO or CR ⁴Wherein be no more than one among V, X, Y and the Z and be NO;

R ^A, R ^A1, R ^BAnd R ^B1Independently be separately: H, OH, halogen, C _1-6Alkyl, C _1-6Alkenyl, C _1-6Alkynyl, C _1-6Haloalkyl, C _1-6Alkoxyl, C _1-6Halogenated alkoxy, heterocyclic radical, carbocylic radical, NR ¹⁰R ¹², NR ¹⁰CO ₂R ¹¹NR ¹⁰CONR ¹⁰R ¹², NR ¹⁰SO ₂NR ¹⁰R ¹², NR ¹⁰-SO ₂-R ¹¹, CN, CONR ¹⁰R ¹², CO ₂R ¹⁰, NO ₂, SR ¹⁰, SOR ¹⁰, SO ₂R ¹⁰Or SO ₂-NR ¹⁰R ¹²

R ¹Be C _1-6Alkyl, C _1-6Haloalkyl, C _1-6Hydroxy alkyl ,-(C _0-6Alkyl)-O-(C _1-6Alkyl) ,-(C _0-6Alkyl)-S-(C _1-6Alkyl) ,-(C _0-6Alkyl)-(C _3-7Cycloalkyl)-(C _0-6Alkyl), OH, OR ¹⁰, SR ¹⁰, COR ¹¹, CO ₂R ¹⁰, CONR ¹⁰R ¹², carbocylic radical, heterocyclic radical, CN, NR ¹⁰R ¹², NR ¹⁰SO ₂R ¹⁰, NR ¹⁰COR ¹⁰, NR ¹⁰CO ₂R ¹⁰, NR ¹⁰CONR ¹², CR ¹⁰R ¹¹CO ₂R ¹⁰Or CR ¹⁰R ¹¹OCOR ¹⁰

R ², R ³, R ⁴, R ⁵And R ⁶Independently be separately: H, OH, halogen, C _1-6Alkyl, C _1-6Haloalkyl, C _1-6Alkoxyl, C _1-6Halogenated alkoxy, C _1-6Thio alkoxy, NR ¹⁰R ¹², NR ¹⁰CO ₂R ¹¹NR ¹⁰CONR ¹⁰R ¹², NR ¹⁰SO ₂NR ¹⁰R ¹², NR ¹⁰-SO ₂-R ¹¹, heterocyclic radical, carbocylic radical, carbon epoxy radicals, heterocyclic oxy group, CN, NO ₂, COR ¹¹, CONR ¹⁰R ¹², CO ₂R ¹⁰, NO ₂, SR ¹⁰, SOR ¹⁰, SO ₂R ¹⁰Or SO ₂-NR ¹⁰R ¹²

R ⁷For H or randomly by individual halogen, OH, the CO of being selected from of 1-3 ₂H, CO ₂-(C _1-6Alkyl) or C _1-3The C that the substituent group of alkoxyl replaces _1-6Alkyl;

R ⁸Be C _1-3Alkoxyl, C _1-3Halogenated alkoxy, C _3-6Cycloalkyloxy or OH;

R ^{8 '}Be H;

R ⁹And R ^{9 '}Independent separately is H, C _1-6Alkyl, halogen, C _1-3Alkoxyl, C _1-3Halogenated alkoxy, C _3-6Cycloalkyl, C _3-6Cycloalkyloxy, OH, CO ₂R ¹⁰, OCOR ¹⁰, wherein said C _1-6Alkyl is randomly by one or more F, C of being selected from _1-3Alkoxyl, OH or CO ₂R ¹⁰Substituent group replace;

Or R ⁹And R ^{9 '}Form 3-7 unit volution base with the carbon atom that they connected;

R ¹⁰Be H, C _1-6Alkyl, benzyl, phenyl or C _3-6Cycloalkyl, wherein said C _1-6Alkyl, benzyl, phenyl or C _3-6Cycloalkyl randomly is selected from halogen, OH, C by 1-3 _1-3Alkyl, C _1-3Haloalkyl, C _1-3Alkoxyl, C _1-3Halogenated alkoxy, CO ₂H, and CO ₂-(C _1-6Alkyl) substituent group replaces;

R ¹¹Be H, OH, C _1-6Alkyl, C _1-6Alkoxyl, benzyl, phenyl, benzyloxy, phenoxy group, C _3-6Cycloalkyl or C _3-6Cycloalkyloxy, wherein said C _1-6Alkyl, C _1-6Alkoxyl, benzyl, phenyl, benzyloxy, phenoxy group, C _3-6Cycloalkyl or C _3-6Cycloalkyloxy randomly is selected from halogen, OH, C by 1-3 _1-3Alkyl, C _1-3Alkoxyl, CO ₂H, CO ₂-(C _1-6Alkyl) and CF ₃Substituent group replace;

R ¹²Be H, C _1-6Alkyl, benzyl, phenyl or C _3-6Cycloalkyl, wherein said C _1-6Alkyl, benzyl, phenyl or C _3-6Cycloalkyl randomly is selected from halogen, OH, C by 1-3 _1-3Alkyl, C _1-3Haloalkyl, C _1-3Alkoxyl, C _1-3Halogenated alkoxy, CO ₂H, and CO ₂-(C _1-6Alkyl) substituent group replaces; And

P is 0 or 1.

The present invention provides the chemical compound of general formula I I especially:

Or its pharmaceutically acceptable salt or prodrug, wherein:

Dotted line is represented the key chosen wantonly;

W is:

X is N, NO or CR ²

Y is N, NO or CR ³

Z is N, NO or CR ⁴Wherein be no more than one among X, Y and the Z and be NO;

R ¹Be C _1-6Alkyl, C _1-6Haloalkyl, (C _0-6Alkyl)-O-(C _1-6Alkyl), (C _0-6Alkyl)-S-(C _1-6Alkyl), (C _0-6Alkyl)-(C _3-7Cycloalkyl)-(C _0-6Alkyl), OH, OR ¹⁰, SR ¹⁰, COR ¹¹, CO ₂R ¹⁰, CONR ¹⁰R ¹², carbocylic radical, heterocyclic radical, CN, NR ¹⁰R ¹², NR ¹⁰SO ₂R ¹⁰, NR ¹⁰COR ¹⁰, NR ¹⁰CO ₂R ¹⁰, NR ¹⁰CONR ¹², CR ¹⁰R ¹¹CO ₂R ¹⁰Or CR ¹⁰R ¹¹OCOR ¹⁰

R ⁸Be C _1-3Alkoxyl, C _1-3Halogenated alkoxy, C _3-6Cycloalkyloxy or OH;

R ^{8 '}Be H;

P is 0 or 1.

In certain embodiments, W is:

In certain embodiments, W is:

In certain embodiments, V is CR ⁵

In certain embodiments, X is CR ²

In certain embodiments, Y is CR ³

In certain embodiments, Z is CR ⁴

In certain embodiments, X is N.

In certain embodiments, Z is N.

In certain embodiments, X and Z are N.

In certain embodiments, X is CR ²Y is CR ³And Z is CR ⁴

In certain embodiments, V is CR ⁵, X is CR ²Y is CR ³And Z is CR ⁴

In certain embodiments, be no more than 2 among V, X, Y and the Z and be N.

In certain embodiments, among V, X, Y and the Z at least 2 be not N or NO.

In certain embodiments, there is not 1 to be N or NO among V, X, Y and the Z.

In certain embodiments, 1 among V, X, Y and the Z is N.

In certain embodiments, 2 among V, X, Y and the Z are N.

In certain embodiments, R ^A, R ^A1, R ^BAnd R ^B1Independent separately is H, OH, halogen, C _1-6Alkyl, C _1-6Alkenyl, C _1-6Alkynyl, C _1-6Haloalkyl, C _1-6Alkoxyl or C _1-6Halogenated alkoxy.

In certain embodiments, R ^A, R ^A1, R ^BAnd R ^B1Independent separately is H, OH or C _1-6Alkoxyl.

In certain embodiments, R ^A, R ^A1, R ^BAnd R ^B1Independent separately is H or OH.

In certain embodiments, R ¹Be C _1-6Alkyl, C _1-6Hydroxy alkyl ,-(C _0-6Alkyl)-O-(C _1-6Alkyl) or heterocyclic radical.

In certain embodiments, R ¹Be C _1-6Alkyl.

In certain embodiments, R ¹It is third-2-base.

In certain embodiments, R ⁵And R ⁶Be not H.

In certain embodiments, R ⁵And R ⁶Be C _1-4Haloalkyl.

In certain embodiments, R ⁶Be C _1-4Haloalkyl.

In certain embodiments, R ⁶Be CF ₃

In certain embodiments, R ⁷Be H.

In certain embodiments, R ⁸Be C _1-3Alkoxyl or C _1-3Halogenated alkoxy.

In certain embodiments, R ⁸Be C _1-3Alkoxyl.

In certain embodiments, R ⁸Be methoxyl group.

In certain embodiments, R ⁸Be ethyoxyl.

In certain embodiments, R ⁹And R ^{9 '}Be H.

In certain embodiments, p is 0.

In certain embodiments, p is 1.

In certain embodiments, chemical compound of the present invention has general formula I a:

In certain embodiments, chemical compound of the present invention has general formula I b, Ic or Id:

In certain embodiments, chemical compound of the present invention has general formula I e or If:

In certain embodiments, chemical compound of the present invention has general formula I g:

In certain embodiments, chemical compound of the present invention has general formula I h or Ii:

Diverse location place has in this manual disclosed the substituent group of The compounds of this invention with the form of group and scope.Specify, the present invention includes each and every seed combination of member in this class combination range.Term " C for example _1-6Alkyl " especially in order to disclose methyl, ethyl, C separately ₃Alkyl, C ₄Alkyl, C ₅Alkyl and C ₆Alkyl.

With regard to once above The compounds of this invention appearred in variable, each variable can be for being selected from the different piece of the Ma Kushi group that defines this variable.For example, have 2 structures that are present in the R group on the same compound simultaneously if described; These 2 R groups can be represented the different piece that is selected from the Ma Kushi group of R definition so.

Further understand, for clarity sake some feature of the present invention of describing in each embodiment of context can also provide with the combining form of single embodiment.On the contrary, the different characteristic of describing in the single embodiment of context for simplicity's sake of the present invention also can be separately or is provided with the form of the sub-portfolio of any appropriate.

The implication of term used herein " alkyl " is meant the saturated hydrocarbyl into straight or branched.The example of alkyl comprises methyl (Me), ethyl (Et), propyl group (for example just-propyl group and isopropyl), butyl (for example just-butyl, isobutyl group, the tert-butyl group), amyl group (for example just-amyl group, isopentyl, neopentyl) etc.Alkyl can contain about 20 of 1-, about 20 of 2-, about 10 of 1-, about 8 of 1-, about 6 of 1-, 1-about 4 or about 3 carbon atoms of 1-.

" alkylidene " used herein refers to divalent alkyl.

" C used herein _2-4Alkylidene " refer to the alkylidene of forming by 2-4 carbon atom.

" alkenyl " used herein refers to the alkyl that has one or more carbon-to-carbon double bonds.Non-limiting examples of alkenyls comprises vinyl, acrylic, cyclohexenyl group etc.

" alkynyl " used herein refers to the alkyl that has one or more carbon-to-carbon triple bonds.The example of alkynyl comprises acetenyl, propinyl etc.

" haloalkyl " used herein refers to the alkyl that has one or more halogenic substituents.The example of haloalkyl comprises CF ₃, C ₂F ₅, CHF ₂, CCl ₃, CHCl ₂, C ₂Cl ₅Deng.

" aryl " used herein refers to monocycle or multi-ring (for example having 2,3 or 4 fused rings) aromatic hydrocarbons, such as, for example phenyl, naphthyl, anthryl, phenanthryl, indanyl, indenyl etc.In certain embodiments, aryl has about 20 carbon atoms of 6-.

" carbocylic radical " used herein is the cyclic hydrocarbon part of saturated (promptly not containing two keys or triple bond) or unsaturated (promptly containing one or more pairs of keys or triple bond).Carbocylic radical can for single-, many-(for example 2,3 or 4 fused rings).The example of carbocylic radical comprises cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, suberyl, cyclopentenyl, 1 base, cyclohexenyl group, norborny, falls pinanyl, norcarnyl, adamantyl, phenyl etc.Carbocylic radical can be for aromatics (for example " aryl ") or non--aromatics (for example " cycloalkyl ").In certain embodiments, carbocylic radical can have about 30 of about 3-, about 20 of about 3-, about 3-about 10 or about 7 the one-tenth ring carbon atoms of about 3-.

" cycloalkyl " used herein refers to non--aromatic carbocyclic class, comprises cyclic alkyl, alkenyl and alkynyl.That cycloalkyl can comprise is single-or multi-ring (for example having 2,3 or 4 fused rings) ring system and spiro ring system.The example of cycloalkyl comprises cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, suberyl, cyclopentenyl, cyclohexenyl group, cyclohexadienyl, cycloheptatriene base, norborny, falls pinanyl, norcarnyl, adamantyl etc.In the definition of cycloalkyl, also comprise and have one or more and part cycloalkyl ring condensed (promptly having the key that has jointly with it) aromatic ring, for example benzo derivative of pentane, amylene, hexane etc.In certain embodiments, cycloalkyl can have about 10 of about 3-, and about 3-Yue 10 or about 3-Yue 7 become ring carbon atom.In certain embodiments, cycloalkyl can have 0,1,2,3,4 or 5 two key or triple bond.In another embodiment, one or more one-tenth ring carbon atoms of cycloalkyl can be replaced by oxo or thio group.

" heterocyclic radical " used herein or " heterocycle " refer to saturated or unsaturated cyclic hydrocarbon, and wherein one or more one-tenth ring carbon atoms of this cyclic hydrocarbon are replaced by hetero atom, such as O, S or N.Heterocyclic radical can be for aromatics (for example " heteroaryl ") or non--aromatics (for example " Heterocyclylalkyl ").Heterocyclic radical can also be equivalent to hydrogenation and partially hydrogenated heteroaryl.That heterocyclic radical can comprise is single-or multi-ring (for example having 2,3 or 4 fused rings) ring system.Possible being characterised in that of heterocyclic radical has 3-14 or 3-7 one-tenth annular atoms.In certain embodiments, heterocyclic radical is except that containing at least one hetero atom, and it is about 13 to contain the 1-that has an appointment, about 7 carbon atoms of about 2-about 10 or about 2-and can provide carbon atom or hetero atom connects.In another embodiment, arbitrarily ring formation carbon or hetero atom can oxidized (for example having oxo or sulfo-substituent group) or nitrogen-atoms can be quaternized.The example of heterocyclic radical comprises morpholino, thiomorpholine generation, piperazinyl, tetrahydrofuran base, tetrahydro-thienyl, 2,3-dihydro benzo furyl, 1,3-benzo dioxole, phendioxin, 4-two  alkane, piperidyl, pyrrolidinyl, different  oxazolidinyl, isothiazole alkyl, pyrazolidinyl,  oxazolidinyl, thiazolidinyl, imidazolidinyl etc. and hereinafter listed any group to " heteroaryl " and " Heterocyclylalkyl ".Heterocyclic additional examples comprises pyrimidine radicals, phenanthridinyl, the phenanthroline base, phenazinyl, phenothiazinyl, phenoxathiinyl, fen  piperazine base, phthalazinyl, piperazinyl, piperidyl, 3,6-dihydropyridine base, 1,2,3, the 6-tetrahydro pyridyl, 1,2,5, the 6-tetrahydro pyridyl, piperidone base, the 4-piperidone base, piperonyl, pteridine radicals, purine radicals, pyranose, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyrido  azoles, pyridine-imidazole, the pyrido thiazole, pyridine radicals (pyridinyl), pyridine radicals (pyridyl), pyrimidine radicals, pyrrolidinyl, pyrrolinyl, the 2H-pyrrole radicals, pyrrole radicals, tetrahydrofuran base, tetrahydro isoquinolyl, tetrahydric quinoline group, tetrazole radical, 6H-1,2,5-thiophene-diazine, 1,2, the 3-thiadiazolyl group, 1,2, the 4-thiadiazolyl group, 1,2, the 5-thiadiazolyl group, 1,3, the 4-thiadiazolyl group, thianthrene group (thianthrenyl), thiazolyl, thienyl, the thieno thiazolyl, thieno  azoles base, the Thienoimidazole base, thiophenyl, triazine radical, 1,2, the 3-triazolyl, 1,2, the 4-triazolyl, 1,2, the 5-triazolyl, 1,3, the 4-triazolyl, xanthyl, the octahydro isoquinolyl, the  di azoly, 1,2,3- di azoly, 1,2,4- di azoly, 1,2,5- di azoly, 1,3,4- di azoly, the  oxazolidinyl,  azoles base, the  oxazolidinyl, quinazolyl, quinolyl, the 4H-quinolizinyl, quinoxalinyl, quininuclidinyl, acridinyl, azocine base (azocinyl), benzimidazolyl, benzofuranyl, benzimidazole thiophanate is for furyl, benzo-thiophenyl, the benzoxazol base, benzothiazolyl, the benzotriazole base, the benzo tetrazole radical, benzisoxa  azoles base, the benzisothiazole base, the benzimidazoline base, methylenedioxyphenyl, morpholinyl, phenanthridinyl, ten-hydrogen quinolyl, 2H, 6H-1,5,2 dithiazine bases, dihydrobenzo [2,3-b] oxolane, furyl, the furazan base, carbazyl, the 4aH-carbazyl, carbolinyl, chromanyl, chromenyl, the cinnolines base, imidazolidinyl, imidazolinyl, imidazole radicals, the 1H-indazolyl, indole thiazolinyl (indolenyl), indolinyl, the indolizine base, indyl, the 3H-indyl, isobenzofuran-base, the isochroman base, iso indazolyl, iso-dihydro-indole-group, isoindolyl, isoquinolyl, isothiazolyl and different  azoles base.Heterocyclic other example comprises azetidine-1-base, 2,5-dihydro-1H-pyrroles-1-base, piperidines-1 base, piperazine-1-base, pyrrolidine-1-base, isoquinolin-2-base, pyridine-1-base, 3,6-dihydropyridine-1-base, 2,3-indoline-1-base, 1,3,4,9-Tetrahydrocarboline-2-base, thieno [2,3-c] pyridine-6-base, 3,4,10,10a-tetrahydrochysene-1H-pyrazolo [1,2-a] indole-2-base, 1,2,4,4a, 5,6-six hydrogen-pyrazolo [1,2-a] quinoline-3-base, pyrazolo [1,2-a] quinoline-3-base, diaza ring-1-in heptan base, 1,4,5,6-tetrahydrochysene-2H-benzo [f] isoquinolin-3-base, 1,4,4a, 5,6,10b-six hydrogen-2H-benzo [f] isoquinolin-3-base, 3,3a, 8,8a-tetrahydrochysene-1H-2-azepine-ring penta [a] indenes-2-base and 2,3,4,7-tetrahydrochysene-1H-azepines-1-base, azepine ring-1-in heptan base.

" heteroaryl " used herein refers to and has at least one heteroatomic annular atoms, such as the aromatic heterocycle of sulfur, oxygen or nitrogen.Heteroaryl comprises that monocycle and multi-ring (for example having 2,3 or 4 fused rings) are.The example of heteroaryl includes but not limited to pyridine radicals, pyrimidine radicals, pyrazinyl, pyridazinyl, triazine radical, furyl (furyl) (furyl (furan yl)), quinolyl, isoquinolyl, thienyl, imidazole radicals, thiazolyl, indyl, pyrrole radicals,  azoles base, benzofuranyl, benzothienyl, benzothiazolyl, different  azoles base, pyrazolyl, triazolyl, tetrazole radical, indazolyl, 1,2,4-thiadiazolyl group, isothiazolyl, benzothienyl, purine radicals, carbazyl, benzimidazolyl, indolinyl etc.In certain embodiments, heteroaryl has about 20 carbon atoms of 1-, and in other embodiments, has about 20 carbon atoms of about 3-.In certain embodiments, heteroaryl contains about 14 of 3-, and about 7 or 5-6 of 3-becomes annular atoms.In certain embodiments, heteroaryl has about 4 of 1-, 1-about 3 or 1-2 hetero atom.

" Heterocyclylalkyl " used herein refers to non--aromatic heterocycle, comprises cyclic alkyl, alkenyl and alkynyl, and wherein one or more become ring carbon atom by hetero atom, replaces such as O, N or S atom.The example of " Heterocyclylalkyl " comprises morpholino, thiomorpholine generation, piperazinyl, tetrahydrofuran base, tetrahydro-thienyl, 2,3-dihydro benzo furyl, 1,3-benzo dioxole, phendioxin, 4-two  alkane, pyridine radicals, pyrrolidinyl, different  oxazolidinyl, isothiazole alkyl, pyrazolidinyl,  oxazolidinyl, thiazolidinyl, imidazolidinyl etc.In the definition of Heterocyclylalkyl, also comprise and have the part that one or more and non-aromatic heterocyclic condense the aromatic ring of (promptly have and have common key with it), for example phthalimide-based, naphthalimido and heterocyclic benzo derivative are such as indole alkene (indolene) and iso-indoles thiazolinyl (isoindolene groups).In certain embodiments, Heterocyclylalkyl has about 20 carbon atoms of 1-, and has about 20 carbon atoms of about 3-in other embodiments.In certain embodiments, Heterocyclylalkyl contains about 14 of 3-, and about 7 or 5-6 of 3-becomes annular atoms.In certain embodiments, Heterocyclylalkyl has about 4 of 1-, 1-about 3 or 1-2 hetero atom.In certain embodiments, Heterocyclylalkyl contains 0-3 two key.In certain embodiments, Heterocyclylalkyl contains two keys of 0-2 or triple bond.

" volution base " used herein refers to 3-14 unit's cycloalkyl or the 3-14 unit Heterocyclylalkyl that the extra loop alkyl that is connected with it or Heterocyclylalkyl have an atom.

" halogen (halo) " used herein or " halogen (halogen) " comprise fluorine, chlorine, bromine and iodine.

" alkoxyl " used herein refers to-the O-alkyl.The example of alkoxyl comprises methoxyl group, ethyoxyl, propoxyl group (for example just-propoxyl group and isopropoxy), tert-butoxy etc.

" thio alkoxy " used herein refers to-the S-alkyl.

" halogenated alkoxy " used herein refers to-the O-haloalkyl.The example of halogenated alkoxy is OCF ₃

" carbon epoxy radicals " used herein refers to-the O-carbocylic radical.

" cycloalkyloxy " used herein refers to-the O-cycloalkyl.

" carbocylic radical alkyl " used herein refers to the alkyl that is replaced by carbocylic radical.

" aralkyl " used herein or " aryl alkyl " refer to the alkyl that is replaced by aryl.

" cycloalkyl-alkyl " used herein refers to the alkyl that is substituted by cycloalkyl.

" heterocyclic radical alkyl " used herein refers to the moieties that is replaced by assorted carbocylic radical.The example of heterocyclic radical alkyl comprises " heteroaryl alkyl " (alkyl that is replaced by heteroaryl) and " Heterocyclylalkyl alkyl " (alkyl that is replaced by Heterocyclylalkyl).In certain embodiments, the heterocyclic radical alkyl has 3-24 carbon atom and becomes ring hetero atom with at least one.

" oxo " used herein refers to=O.

Chemical compound as herein described can be asymmetric (for example having one or more three-dimensional centers).Except as otherwise noted, refer to all stereoisomers, such as enantiomer and diastereomer.The The compounds of this invention that can separate the carbon atom that contains asymmetric replacement of optically-active or racemic form.How the method by optically-active feedstock production optically-active form is as known in the art, such as by resolving racemic mixtures or synthetic by stereo selectivity.The geometric isomer of the two keys of many olefines, C=etc. also may reside in the chemical compound as herein described, and the present invention expects the isomer that all these classes are stable.The cis of having described The compounds of this invention is with trans geometric isomer and they can be separated as isomer mixture or as independent isomeric forms.

Can split the racemic mixture of chemical compound by big metering method as known in the art.Case method comprises that the salify organic acid " chiral separation acid " that uses to optically-active carries out fractional recrystallization.The suitable resolution reagent that is used for the fractional recrystallization method is: for example optically-active acid, and such as tartaric acid, diacetyl tartaric acid, dibenzoyl tartaric acid, mandelic acid, malic acid, lactic acid or various optically-active camphorsulfonic acid, such as the D and the L shaped formula of beta camphor sulfonic acid.Other resolution reagent that is suitable for the fractional recrystallization method comprises the pure form of stereoisomer of Alpha-Methyl benzylamine (for example S and R form or the pure form of non-stereoisomer), 2-phenyl glycinol, cathine, ephedrine, N-methylephedrine, cyclohexyl ethamine, 1,2-diamino-cyclohexane etc.

Can also be by eluting resolving racemic mixtures on the post of having filled optical resolution reagent (for example dinitrobenzoyl phenylglycine).Those skilled in the art can determine the composition of suitable eluting solvent.

Chemical compound of the present invention also comprises the tautomeride form, such as the ketoenol tautomerization body.

Chemical compound of the present invention can also comprise all isotopes that appear at the atom on intermediate or the final chemical compound.Isotope comprises that those have the same atoms number, but the different atom of atomic weight.For example, the isotope of hydrogen comprises tritium and deuterium.

Term used herein " pharmaceutically acceptable " refers to and be applicable to contact humans and animals tissue in sound medical judgment scope, but do not have excessive toxicity, zest, anaphylaxis or other problem or a complication with rational interests/risk than those chemical compounds, material, compositions and/or the dosage form that are complementary.

The present invention also comprises the pharmaceutically acceptable salt of chemical compound described herein." pharmaceutically acceptable salt " used herein refers to the derivant that discloses chemical compound, wherein partly changes into its salt form modification parent compound by the acid or the alkali that will exist.The example of pharmaceutically acceptable salt includes, but are not limited to: alkaline residue, such as the mineral acid or the acylate of amine; Required residue is such as the alkali metal of carboxylic acid or acylate etc.Pharmaceutically acceptable salt of the present invention for example comprises the non-toxic salts or the quaternary ammonium salt of the parent compound that is formed by avirulence mineral acid or organic acid.Can be by the conventional chemical method by the synthetic pharmaceutically acceptable salt of the present invention of the parent compound that contains alkalescence or acidic moiety.In general, can be by free acid or alkali form and stoichiometric suitable alkali or acid this class salt of prepared in reaction in water or organic solvent or their both mixture that makes these chemical compounds; In general, preferred non-aqueous media is as ether, ethyl acetate, ethanol, isopropyl alcohol or acetonitrile.The example of suitable salt can be at Remington ' sPharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p.1418 with Journal of Pharmaceutical Science, find in 66,2 (1977), these documents intactly are incorporated herein by reference separately.

The present invention also comprises the prodrug of chemical compound described herein." prodrug " used herein refers to any covalently bound carrier that discharges active parent drug to the mammalian subject administration time.Can prepare prodrug by the functional group that modification is present on the chemical compound, according to this class mode so that with routine operation or in vivo trim is cracked into parent compound.Prodrug comprises chemical compound, and wherein to the mammalian subject administration time, hydroxyl, amino, sulfydryl or carboxyl combine with any group that can be cracked into free hydroxyl group, amino, sulfydryl or carboxyl respectively.The example of prodrug includes but not limited to acetas, formic acid esters and the benzoate derivatives of the alkohol and amine functional group on the The compounds of this invention.At T.Higuchi and V.Stella, Vol. and the Bioreversible Carriers in Drug Design of " Pro-drugs as Novel Delivery Systems " A.C.S.Symposium Series, ed.Edward B.Roche, American Pharmaceutical Association and Pergamon Press, the preparation and the application of prodrug have been discussed in 1987, the full content of these two pieces of documents has been incorporated herein by reference.

Synthetic

Known organic synthesis technology preparation be can use and can its salt, hydrate and solvate be comprised according to a large amount of possible synthesizing arbitrarily by way of synthetic chemical compound of the present invention.

Can in the suitable solvent that organic synthesis field those of ordinary skill is easy to select, be prepared the reaction of The compounds of this invention.Suitable solvent can not react under the temperature of reacting with raw material (reactant), intermediate or product basically, for example the temperature from the freezing temperature of solvent to solvent boiling temperature scope.Can in solvent or more than one solvent mixtures, carry out specified reaction.According to the difference of specific reactions steps, can select to be used for the suitable solvent of specific reactions steps.

The preparation of The compounds of this invention can comprise the protection of various chemical groups and deprotection.The selection of the requirement of protection and deprotection and suitable protecting group is easy to be determined by those skilled in the art.For example, the protecting group chemistry can be at T.W.Greene and P.G.M.Wuts, ProtectiveGroups in Organic Synthesis, 3rd.Ed., Wiley ﹠amp; Sons, Inc., NewYork finds in (1999), and the full content of the document is incorporated herein by reference.

Can be according to the method monitoring reaction of any appropriate well known in the art.For example, can pass through spectrophotography, (for example such as nuclear magnetic resonance spectrometry ¹H or ¹³C), infrared spectrometry, spectrophotometry (for example UV-visible light) or mass spectrography or by chromatography, such as high performance liquid chromatography (HPLC) or thin layer chromatography monitoring reaction.

Provide among the scheme 1-13 hereinafter the typical case of The compounds of this invention synthetic by way of, the formation member of wherein said general formula is as defined herein.

Scheme described in can operational version 1 prepares the 3-aminopentane carboxylic acid of general formula 1-5.Can be by in DMF, the carboxylic acid 1-1 that is purchased being changed into ester, such as methyl ester with iodomethane/potassium carbonate processing.Can use alkali, such as hexamethyl two silicon lithium nitrides (LHMDS) make gained ester 1-2 use halogenide, such as iodide (R ¹I) alkylation and alkylate 1-3 is provided, it is the mixture (4: 1 ratios) of cis and trans diastereomer.Can remove a spot of trans enantiomer by after ester is hydrolyzed into acid, carrying out crystallization.Can use catalyst, make the sour 1-4 of gained enantiomer-pure carry out hydrogenation and obtain saturated carboxylic acid 1-5 such as Pd-C.

Scheme 1

Generalized operating procedure prepares the cyclopentane-carboxylic acid of general formula 2-5 in can operational version 2.The 3-oxo-cyclopentane formic acid 2-1 that is purchased can be changed into ester, such as methyl ester.Can be by acidic catalyst (catlyst) being arranged, handling the ketone of protecting gained ester 2-2 with trimethyl orthoformate in the presence of such as p-methyl benzenesulfonic acid.Can use alkali, use alkyl iodide (R such as LHMDS ¹The alkylation of the ketal 2-3 of generation I).Use alkali, make the 2-4 hydrolysis of alkylation ester and obtain the carboxylic acid of general formula 2-5 such as LiOH, NaOH or KOH.

Scheme 2

Operating procedure described in the operational version 3 prepares bridged piperazine derivatives.Use Hydro-Giene (Water Science). (I) and potassium phosphate to make the iodobenzene derivant coupling of the bridged piperazine derivatives of general formula 3-2 and general formula 3-1 and obtain intermediate 3-3.Use acid, remove the Boc group and obtain the bridged piperazine derivatives of general formula 3-4 such as the HCl in two  alkane or TFA.

Scheme 3

Perhaps, can prepare bridged piperazine derivatives (general formula 4-3) by 2-chloropyridine or the 2-chloropyrimide derivant that replaces general formula 4-1 with the bridged piperazine derivatives of general formula 4-2.

Scheme 4

Perhaps, the order of explaining in can operational version 5 prepares bridged piperazine derivatives.Can by will be purchased with bromination isopropyl-magnesium and iodinate 3,5-dibromo pyridine 5-1 changes into 3-bromo-5-iodine pyridine 5-2.Can use Hydro-Giene (Water Science). (I) and potassium phosphate to make the bridged piperazine derivatives coupling of gained iodine and general formula 3-2.After the bromine that uses bromination isopropyl-magnesium and iodine with gained intermediate 5-3 changes into iodine, can be by using Me ₃SiCF ₃/ CuI/KF/DMF handles the 5-flumethiazine derivant that obtains general formula 5-5 with trifluoromethyl replacement iodine.Use acid, remove Boc and obtain the bridged piperazine derivatives of general formula 5-6 such as the HCl in two  alkane or TFA.

Scheme 5

Can as shown in scheme 6, synthesize piperidines or 5,6-tetrahydropyridine derivative.Use lithium alkylide, such as just-butyl lithium or tert-butyl lithium make bromo-or the iodobenzene derivant lithiumation of general formula 6-1, obtain the tertiary alcohol of general formula 6-3 subsequently with the ketone derivatives quencher of general formula 6-2.Using dehydrant, after thionyl chloride/pyridine dehydration, can be by using catalyst, such as Pd/ hydrocarbonize reduction gained alkene.Use acid, handle 6-3,6-4 and 6-5 and obtain the chemical compound of general formula 6-6,6-7 and 6-8 such as the HCl in two  alkane or TFA.

Scheme 6

Perhaps, order synthesizing piperazine or the 5,6-tetrahydropyridine derivative explained in can operational version 7.Can be by using BrSiMe ₃Processing is with the 2-chloropyridine of the general formula 4-1 that is purchased or the 2-bromopyridine derivant that the 2-chloropyridine derivative changes into general formula 7-1.Can use the piperidines and the 5,6-tetrahydropyridine derivative that obtain general formula 7-5 and 7-6 to operating procedure similar described in the scheme 6 by 7-1.

Scheme 7

Perhaps, can be as generalized synthetic piperidines or 5,6-tetrahydropyridine derivative in the scheme 8.Can obtain 3-nitro-5-5-flumethiazine-2-alcohol by the nitrated 5-5-flumethiazine that is purchased-2-alcohol (8-1).After the hydroxyl on the 8-2 is changed into chlorine, use catalyst, make gained chlorine compound 8-3 carry out hydrogenation and obtain 3-amino-5-5-flumethiazine 8-4 such as Pd/ carbon.Use NaNO in the presence of Cu (I) Br is being arranged ₂/ HBr makes 8-4 diazotising and obtains 3-bromo-5-5-flumethiazine 8-5.After the operating procedure described in the scheme 6,8-5 can be changed into piperidines or the 5,6-tetrahydropyridine derivative of general formula 8-9 and 8-10.

Scheme 8

Can be as the acquisition tetrahydropyran derivatives that describes in detail in the scheme 9 (R wherein ^8aFor example be alkyl).Can by be used in the methanol between-the chlorine benzylhydroperoxide handles the 4-methoxyl group-3 that is purchased, 6-dihydro-2H-pyrans 9-1 changes into 4,4-dimethoxy tetrahydrochysene-2H-pyrans-2-alcohol.Use NaH, make the 9-2 alkylation and obtain tri-alkoxy intermediate 9-3 with alkyl halide.Use acid, obtain the ketone product of general formula 9-4 such as HCl aqueous solution processing 9-3.Can carry out the amine that hydrogenation changes into ketone 9-4 general formula 9-6 subsequently by carrying out reductive amination with the ADP methylmethane.

Scheme 9

The final chemical compound of the method combination general formula I described in can operational version 10.Can use standard amide to form reagent, make the amine condensation of carboxylic acid and the general formula 10-1 of general formula 1-5 such as BOP or PyBrop (coupling reagent).Using acid, remove Boc such as HCl or TFA after, use Reducing agent, make the ketone of gained amine 10-3 and general formula 10-4 carry out reductive amination and obtain the final chemical compound of general formula 10-5 such as sodium triacetoxy borohydride.

Scheme 10

Perhaps, can be according to scheme 11 combinations chemical compound of the present invention.Use the standard amide forming method to make the amine coupling of the carboxylic acid of general formula 2-5 and general formula 10-1 and obtain the amide of general formula 11-1.After using sour water that ketal is changed into ketone, use Reducing agent, make the amine of gained ketone 11-2 and general formula 11-3 carry out reductive amination and obtain the chemical compound of general formula 11-4 such as sodium triacetoxy borohydride.

Scheme 11

Method

In certain embodiments, chemical compound of the present invention can be regulated the activity of one or more chemokine receptors.Term " adjusting " refers to the ability that increases or reduce receptor active.Therefore, chemical compound of the present invention can be used for regulating the method for chemokine receptors, contacts any one or multiple carrying out in chemical compound as herein described or the compositions by making described receptor.In certain embodiments, chemical compound of the present invention can be used as inhibitors of chemokine receptors and works.In extra embodiment, chemical compound of the present invention can be used to regulate the chemokine receptor activity that needs are regulated the individuality of receptor, is undertaken by the compound of Formula I that gives regulated quantity.

The compounds of this invention combination and/or the chemokine receptors of regulating comprise chemokine receptors arbitrarily.In certain embodiments, chemokine receptors belongs to the CC family of chemokine receptors, comprising: for example CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8 and CCR10.In certain embodiments, chemokine receptors is CCR2.In certain embodiments, chemokine receptors is CCR5.In certain embodiments, chemokine receptors combination and/or adjusting CCR2 and CCR5.

Chemical compound of the present invention can be for optionally.So-called " selectivity " refers to chemical compound and compares with at least a extra chemokine receptors to have big affinity or effect combination or chemokine inhibiting receptor respectively.

Chemical compound of the present invention is the double inhibitor of CCR2 and CCR5, be chemical compound of the present invention with compare respectively bigger affinity of other chemokine receptors or effect in conjunction with or suppress CCR2 and CCR5, described other chemokine receptors is such as CCR1, CCR3, CCR4, CCR6, CCR7, CCR8 and CCR10.In certain embodiments, chemical compound of the present invention combination that CCR2 and CCR5 are had or suppress selectivity and surpassed the combination of other chemokine receptors arbitrarily or suppress selectivity.Selectivity can be at least about 10-times, at least about 20-times, at least about 50-doubly, at least about 100-doubly, at least about 200-doubly, at least about 500-times or at least about 1000-times.Can be according to the conventional method in this area, such as according to test determination binding affinity provided herein and inhibitor effect.

The present invention further provides the method for individual (for example patient) chemokine receptors-relevant disease of treatment or obstacle, undertaken by The compounds of this invention or its pharmaceutical composition of the individuality of this class treatment of needs being treated effective dose or dosage.Chemokine receptors-relevant disease can comprise directly or indirectly and chemokine receptor expression or relevant any disease, obstacle or the disease of activity.Chemokine receptors-relevant disease can also comprise can be by regulating any disease, obstacle or the disease that chemokine receptor activity can prevent, improves or cure.Chemokine receptors-relevant disease may further include and be characterised in that pathogen (infectious agent), such as virus or virus protein and the bonded any disease of chemokine receptors, obstacle or disease.In certain embodiments, chemokine receptors-relevant disease is the CCR5-relevant disease, infects such as HIV.

The example of chemokine receptors-relevant disease, disease and illness comprises inflammatory and inflammatory diseases, immune disorders, cancer and viral infection.The example of inflammatory diseases comprises: have the disease of inflammation composition, such as asthma, seasonal and allergic rhinitis all the year round, sinusitis, conjunctivitis, the macula lutea degenerative change relevant with the age, food anaphylaxis, scombroid poisoning, psoriasis, urticaria, pruritus, eczema, inflammatory bowel, thrombotic disease, otitis media, liver cirrhosis, heart disease, Alzheimer, sepsis, restenosis, atherosclerosis, multiple sclerosis, Crohn disease, ulcerative colitis, allergy pneumonopathy, drug induced pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), rheumatoid arthritis and nephritis, ulcerative colitis, atopic dermatitis, apoplexy, acute nerve injury, sarcoidosis, hepatitis, endometriosis, neuropathic pain, hypersensitivity pneumonitis, the eosinophilic pneumonia, delayed hypersensitivity, interstitial lung disease (ILD) (for example idiopathic pulmonary fibrosis or the ILD relevant with rheumatoid arthritis, systemic lupus erythematosus (sle), ankylosing spondylitis, systemic sclerosis, xerodermosteosis, polymyositis or dermatomyositis), ophthalmic (retinal degeneration for example, choroidal neovascularization formation etc.) etc.The example of immune disorders comprises rheumatoid arthritis, arthritic psoriasis, systemic lupus erythematosus (sle), myasthenia gravis, juvenile onset diabetes; Glomerulonephritis, autoimmune throiditis, organ-graft refection comprise allograft rejection and graft versus host disease.The example of cancer comprises: cancer such as breast carcinoma, ovarian cancer, multiple myeloma etc., is characterized in that (for example tumor-associated macrophages TAMs) is impregnated into tumor or illing tissue to macrophage.The example of viral infection comprises that herpes infection, HIV infect or AIDS.

The specified portions that term used herein " contact " refers in vitro system or the interior system of body contacts with each other.For example, individuality that chemokine receptors " contact " chemical compound of the present invention is comprised have chemokine receptors or patient, such as people's chemical compound of the present invention, and, for example chemical compound of the present invention is imported the purification goods that contain the sample of cell or contain chemokine receptors.

Term used herein " individuality " or " patient " can exchange use, refer to any animal, comprise mammal, preferred mice, rat, other rodent, rabbit, Canis familiaris L., cat, pig, cattle, sheep, horse or primates, and optimum is chosen.

Term used herein " treatment effective dose " refers to the reactive compound that causes biology or drug reaction or the amount of pharmaceutically active agents, research worker, veterinary, doctor or other clinicist think that described biology or drug reaction have meaning in tissue, system, animal, individuality or human body, comprise in following one or more:

(1) prevent disease; For example, prevention may susceptibility to disease, disease or obstacle, and (limiting examples is for prevention allergy pneumonopathy, drug induced pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), graft versus host disease and/or the allograft rejection after transplanting but disease, disease or the obstacle of the individuality of disease pathologic condition or symptom do not take place as yet or show; Or Polyglucan reaction, such as atopic dermatitis, delayed hypersensitivity or seasonal or allergic rhinitis all the year round);

(2) suppress disease; For example, disease, disease or the obstacle (promptly stoping pathologic condition and/or symptom to further develop) of the individuality of the pathologic condition of disease, disease or obstacle or symptom suppress to take place or show, such as the inflammation or the autoimmune response that suppress in allergy pneumonopathy, drug induced pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), rheumatoid arthritis, lupus or the psoriasis; Or suppress tumor growth or make the virus load in the viral infection situation keep stable; With

(3) improve disease; Disease, disease or the obstacle (promptly reversing pathologic condition and/or symptom) of the individuality of the pathologic condition of disease, disease or obstacle or symptom for example improve to take place or show, such as the autoimmune response that alleviates in allergy pneumonopathy, drug induced pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), rheumatoid arthritis, lupus or the psoriasis; Or the tumor relevant with cancer shunk back or reduce virus load in the viral infection situation.

The pharmaceutically active agents that one or more are extra, such as, for example antibody, anti-inflammatory agent, immunosuppressant, chemotherapeutics can be united with The compounds of this invention and be used for the treatment of chemokine receptors-relevant disease, obstacle or disease.These activating agents and The compounds of this invention can be incorporated in the single dosage form, maybe can be with these activating agents as independent dosage form while or administration successively.

The pharmaceutically active agents that one or more are extra, such as, for example antiviral agent, antibody, anti-inflammatory agent, Drugs Promoting Insulin Secretion and sensitizer, serum lipids and lipid carrier regulator, and/or immunosuppressant can be united with The compounds of this invention and is used for the treatment of chemokine receptors-relevant disease, obstacle or disease.Can with these activating agents and The compounds of this invention is incorporated in single or continuous dosage form in, maybe can be as independent dosage form simultaneously or administration successively with these activating agents.

The suitable antiviral agents of being paid close attention to the The compounds of this invention coupling can comprise nucleoside and nucleotide reverse transcriptase inhibitors (NRTIs), protease inhibitor and other antiviral agents.

The suitable antiviral agents of being paid close attention to the The compounds of this invention coupling can comprise nucleoside and nucleotide reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitor, entry inhibitor, fusion inhibitor, ripe inhibitor and other antiviral agents.

Suitable NRTIs example comprises: zidovudine (AZT); Didanosine (ddl); Zalcitabine (ddC); Stavudine (d4T); Lamivudine (3TC); Abacavir (1592U89); Adefovir dipivoxil [two (POM)-PMEA]; Lobucavir (BMS-180194); BCH-10652; Emtricitabine [(-)-FTC]; β-L-FD4 (be also referred to as β-L-D4C and be called β-L-2 ', 3 '-two deoxidations (dicleoxy)-5-fluoro-cytidine (cytidene)); DAPD, ((-)-β-D-2,6 ,-diaminourea-purine dioxolane); And lodenosine (FddA).

Typical suitable NNRTIs comprises: nevirapine (BI-RG-587); Delavirdine (BHAP, U-90152); Efavirenz (DMP-266); PNU-142721; AG-1549; MKC-442 (1-(ethyoxyl-methyl)-5-(1-Methylethyl)-6-(phenyl methyl)-(2,4 (1H, 3H)-hybar X); (+)-calanolide A (calanolide) A (NSC-675451) and B.

Typical suitable protease inhibitor comprises: Saquinavir (Ro 31-8959); Ritonavir (ABT-538); Indinavir (MK-639); Viracept see nelfinaivr (nelfnavir) (AG-1343); Amprenavir (141W94); LASINAVIR BMS-234475 Lasinavir [INN (BMS-234475); DMP-450; BMS-2322623; ABT-378; With AG-1 549.

Other antiviral agents comprises hydroxyurea, ribavirin, IL-2, IL-12, pentafuside, enfuvirtide, C-34, ring three azepine sulfonamide CADA, PA-457 and Yissum ProjectNo.11607.

In certain embodiments, with the The compounds of this invention coupling various anti-inflammatory agent or analgesic can comprise: opiate agonist for example; Lipoxidase inhibitor is such as the 5-lipoxidase inhibitor; Cyclooxygenase-2 inhibitor is such as cyclooxygenase-2 inhibitor; Interleukin inhibitors is such as the interleukin-1 inhibitor; Tnf inhibitor is such as infliximab, Embrel or adalimumab; The NNMA antagonist; Nitric oxide inhibitor or nitric oxide synthetic inhibitor; The anti-inflammatory agent of nonsteroidal antiinflammatory agent or inhibition cytokine, for example, such as acetaminophen, aspirin, codeine, fentanyl, ibuprofen, indomethacin, ketodolac, morphine, naproxen, phenacetin, piroxicam, steroid analgesic, sufentanil, sulindac (sunlindac), tenidap etc.Similarly, can be with chemical compound of the present invention with following medicine administration: pain relief agents; Synergist is such as caffeine, H2-antagonist, simethicone, aluminium hydroxide or magnesium hydroxide; Decongestant is such as phenylephrine, phenylpropanolamine, pseudoephedrine, oxymetazoline, ephinephrine, naphazoline, xylometazoline, propylhexedrine or left-handed-metamfetamine; Antfitussive is such as codeine, hydrocodone, caramiphen, carbetapentane or dextramethorphan; Diuretic; With calmness or non-sedative antihistamine medicine.

In certain embodiments, the pharmaceutically active agents of being paid close attention to the The compounds of this invention coupling can include but not limited to: (a) VLA-4 antagonist, such as being described in US 5,510,332, those among WO95/15973, WO96/01644, WO96/06108, WO96/20216, WO96/229661, WO96/31206, WO96/4078, WO97/030941, WO97/022897, WO98/426567, WO98/53814, WO98/53817, WO98/538185, WO98/54207 and the WO98/58902; (b) steroid is such as beclometasone (beclornethasone), methylprednisolone (methyl pi-ednisolone), betamethasone (betarnethasone), prednisone, dexamethasone and hydrocortisone; (c) immunosuppressant is such as Cyclosporin A, tacrolimus, rapamycin (raparnycin) and other FK506 para-immunity inhibitor; (d) antihistaminic (HI-histamine antagonist) is such as brompheniramine (bromopheniramine), chlorphenamine, dexchlorpheniramine, triprolidine, clemastine, diphenhydramine, diphenylpyraline, tripelennamine, hydroxyzine, methdilazine, promethazine, alimemazine, azatadine, Cyproheptadine, antazoline, pheniramine, pyrilamine (pyrilarnine), astemizole (asternizole), terfenadine, loratadine, cetirizine, fexofenadine, desearboethoxyloratadine etc.; (e) non-steroid antiasthmatics, such as terbutaline, orciprenaline, fenoterol, isoetarine (isoethaiine), albuterol, bitolterol, pirbuterol, bitter edible plant alkali, sodium cromoglicate, atropine, ipratropium bromide, leukotriene antagonist (for example zafirlukast, montelukast, pranlukast, iralukast, pobilukast, SKB-106,203), leukotriene biosynthesis inhibitor (for example zileuton, BAY-1005); (f) on-steroidal AID (NSAIDs) is such as propanoic derivatives (alminoprofen for example, benzene  Lip river sweet smell, the bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen, pranoprofen, suprofen, tiaprofenic acid and sulfur  Lip river sweet smell), acetogenin (indomethacin for example, acemetacin (acernetacin), alclofenac, clidanac, diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac, Isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin and zomepirac), fenarnic acid derivant (flufenamic acid (flufenarnic acid), meclofenamic acid, mefenamic acid (rnefenamicacid), niflumic acid and tolfenarnic acid), biphenyl carboxylic acids (biphenylearboxylicacid) derivant (diflunisal and flufenisal), former times health class (oxicarns) (isoxicam (isoxicarn), piroxicam, sudoxicam and tenoxicam), salicylic acid (aspirin, sulfasalazine) and pyrazolone (azapropazone, benzpiperilone (bezpiperylon), feprazone, mofebutazone, oxyphenbutazone, Phenylbutazone); (g) cyclo-oxygenase-2 (COX-2) inhibitor; (h) phosphodiesterase IN type (PDE-IV) inhibitor; (i) other antagonist of chemokine receptors, especially CXCR-4, CCR1, CCR2, CCR3 and CCR5; (j) pravastatin is such as HMG-CoA reductase inhibitor (lovastatin, sirrivastatin and pravastatin, fluvastatin, atorvastatin and other statins), sequestering agent (colestyramine and colestipol), nicotinic acid, fenofibric acid derivant (gemfibrozil, clofibrate, fenofibrate and bezafibrate); And probucol; (k) antiinflammatory biological preparation is such as anti-TNF therapy, anti--the IL-1 receptor, CTLA-4Ig, anti-CD 20 and anti--VLA4 antibody; (l) antidiabetic drug is such as insulin, sulfonylurea, biguanides (metformin), U.-glycosidase inhibitor (acarbose) and orlitazones (troglitazone and pioglitazone); (m) interferon beta goods (interferon beta-1o., interferon beta-1P); (n) other chemical compound is such as the aminosallcylic acid class; Antimetabolite is such as azathioprine and 6-mercaptopurine and cytotoxicity cancer chemotherapeutic agent.The weight ratio of The compounds of this invention and second kind of active component can change and depend on the effective dose of every kind of component.

For example, CCR2 and/or CCR5 antagonist can be united with the anti-inflammatory drug activating agent and be used for the treatment of inflammation, metabolic disease, autoimmune disease, cancer or viral infection, so that improve therapeutic response with the reacting phase ratio of independent therapeutic agent, but can not aggravate toxic action.Additional or synergism is the desired result of CCR2 of the present invention and/or CCR5 antagonist and extra activating agent coupling.In addition, cancerous cell can obtain reversing when using CCR2 of the present invention and/or CCR5 antagonist for treating to the toleration such as this class activating agent of dexamethasone.

Pharmaceutical preparation and dosage form

When as drug use, can give the chemical compound of general formula I with the form of pharmaceutical composition.Can prepare these compositionss according to the well-known mode of pharmaceutical field, and can be according to being that needs topical therapeutic or whole body therapeutic and the area of being treated give them by different approaches.Administration can be local (comprise eye and to mucosa delivery, comprise intranasal, vagina and rectum send); Pulmonary (for example by sucking or being blown into powder or aerosol, comprises the use aerosol apparatus; In the trachea, intranasal, epidermis and transdermal), oral or non-intestinal.Parenterai administration comprises: intravenous, subcutaneous, intraperitoneal, intramuscular or injection or infusion; Or intracranial, for example sheath is interior or the interior administration of ventricle.Parenterai administration can be disposable bolus dose form, or for example can pass through the administration of continous pouring pump.The pharmaceutical composition and the preparation that are used for topical can comprise transdermal patch, ointment, lotion, gel, drop, suppository, spray, liquid and powder.Pharmaceutical carrier, water, powder or oil matrix, thickening agents etc. commonly used may be necessary or need.The condom of coating, glove etc. also may be useful.

The present invention also comprises pharmaceutical composition, and they contain the chemical compound of above-mentioned one or more general formula Is as active component and one or more pharmaceutically acceptable carriers.In the process of the preparation present composition, generally with active component and mixed with excipients, with excipient dilution or be encapsulated in such as, for example in the carrier of capsule, sachet, paper or other vessel form.When excipient was used as diluent, it can be solid, semisolid or liquid substance, plays vehicle, carrier or the medium of active component.Therefore, compositions can be for tablet, pill, powder, lozenge, sachet, cachet, elixir, suspension, Emulsion, solution, syrup, aerosol (as solid or in liquid medium), contain the form of the ointment that for example reaches 10% reactive compound weight, soft hard capsule, suppository, sterile injectable solution and aseptic packaging powder.

In the process of preparation preparation, reactive compound can be ground into suitable granular size, after this merge with other component.If it is insoluble that reactive compound is essentially, it can be ground into so and be lower than 200 order granular sizes.If it is water miscible that reactive compound is essentially, can adjust granular size, for example about 40 orders by being ground in preparation equally distributed basically granular size so.

Some example of suitable excipient comprises lactose, glucose, sucrose, sorbitol, mannitol, starch, Radix Acaciae senegalis, calcium phosphate, alginate, Tragacanth, gelatin, calcium silicates, microcrystalline Cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose.Preparation can also comprise: lubricant, such as Pulvis Talci, magnesium stearate and mineral oil; Wetting agent; Emulsifying agent and suspending agent; Antiseptic is such as methyl-and propyl hydroxy-benzoates; Sweetener; And correctives.Compositions of the present invention can be mixed with to make it discharging active component by quick, lasting after using operating procedure well known in the art to patient's administration or delay.

Compositions can be mixed with unit dosage forms, each dosage contains the about 1000mg of the 5-that has an appointment (1g), more generally be about the about 500mg active component of 100-.Term " unit dosage forms " refers to the physics discrete units that is suitable for as human body experimenter and other mammiferous unit dose, and constituent parts dosage contains the required therapeutical effect of promising generation and the active substance of the scheduled volume that calculates and suitable drug excipient.

In certain embodiments, chemical compound of the present invention or compositions contain the about 50mg active component of the 5-that has an appointment.It will be appreciated by those skilled in the art that it can be presented as that to contain the 5-that has an appointment about 10, about 10-is about 15, and about 15-is about 20, and about 20-is about 25, and about 25-is about 30, and about 30-is about 35, and about 35-is about 40, the chemical compound or the compositions of about 40-about 45 or the about 50mg active component of about 45-.

In certain embodiments, chemical compound of the present invention or compositions contain the about 500mg active component of the 50-that has an appointment.It will be appreciated by those skilled in the art that it can be presented as that to contain the 50-that has an appointment about 75, about 75-is about 100, and about 100-about 125, about 125-is about 150, and about 150-is about 175, and about 175-about 200, about 200-is about 225, and about 225-is about 250, and about 250-about 275, about 275-is about 300, and about 300-is about 325, and about 325-about 350, about 350-is about 375, and about 375-is about 400, and about 400-about 425, about 425-is about 450, the chemical compound or the compositions of about 450-about 475 or the about 500mg active component of about 475-.

In certain embodiments, chemical compound of the present invention or compositions contain the about 1000mg active component of the 500-that has an appointment.It will be appreciated by those skilled in the art that it can be presented as contains the 500-that has an appointment about 550, about 550-about 600, about 600-about 650, about 650-about 700, about 700-is about 750, and about 750-is about 800, and about 800-about 850, about 850-is about 900, the chemical compound or the compositions of about 900-about 950 or the about 1000mg active component of about 950-.

Reactive compound can be in wide dosage range effectively and generally with the medicine effective quantity administration.Yet, the actual dosage that is appreciated that chemical compound is determined according to correlation circumstance by the clinicist usually, comprises age, body weight and the reaction of the disease of being treated, the route of administration of selection, actual administered compound, individual patient, the order of severity of patient's symptom etc.

In order to prepare solid composite,, main active component and drug excipient are mixed into the solid preformulation composition of the homogeneous mixture that contains The compounds of this invention such as tablet.When mentioning that these preformulation composition are even, active component generally is evenly dispersed in the whole compositions, makes to be easy to said composition is divided into equivalent unit dosage forms again, such as tablet, pill and capsule.Then this solid preformulation agent is divided into again and contains for example unit dosage forms of the above-mentioned type of the about 1000mg of 0.1-active component of the present invention.

Can be with tablet of the present invention or coating of pill, otherwise just they are mixed into the dosage form that prolongation effect advantage can be provided.For example, tablet or pill can comprise internal layer dosage and outer dose components, and the latter is the shell form on the former.Two kinds of compositions can be separated to come by enteric layer, and enteric layer is used to stop disintegrate under one's belt and enters duodenum with allowing the internal layer complete components or postpone release.Various materials can be used for this class enteric layer or coatings, and this class material comprises number of polymers acid and polymeric acid and such as this class mixtures of material of Lac, spermol and cellulose acetate.

Can be used for oral or comprise aqueous solution, suitably syrup, water or oil suspension and the use edible oil of flavoring, such as the Emulsion of Oleum Gossypii semen, Oleum sesami, Oleum Cocois or Oleum Arachidis hypogaeae semen flavoring and elixir and similar pharmaceutical carrier by the liquid form that drug administration by injection is introduced The compounds of this invention and compositions.

The compositions that is used for sucking or be blown into is included in solution and the suspension and the powder of pharmaceutically acceptable water or organic solvent or its mixture.The liquid or solid compositions can contain suitable as mentioned above pharmaceutically acceptable excipient.In certain embodiments, by oral or nasal respiration by way of giving compositions so that produce part or general action.Can be by using the inert gas atomizer compositions.Can directly suck atomized soln from sprayer unit maybe can make sprayer unit be connected with face shield tent or intermittent positive pressure breathing machine.Can be to give solution, suspension or powder composition by oral cavity or nose the device of suitable method delivery formulation.

To the amount of patient's administered compound or compositions according to giving which kind of medicine, the administration purpose, such as prevention or treatment, the difference of patient's states, administering mode etc. and changing.In treatment is used, can be enough to cure or stop the compositions of the consumption of disease symptoms and complication thereof to ill patient to small part.Effective dose depends on that the clinicist of the disease situation of being treated and participation is according to the judgement such as this class factors such as disease severity, patient age, body weight and ordinary circumstances.

The compositions that the patient is given can be the aforementioned pharmaceutical compositions form.Can maybe they can be carried out aseptic filtration to these compositions sterilizations by conventional aseptic technique.Can wherein before administration, lyophilized formulations and aseptic aqueous carrier be merged according to the Different Package aqueous solution of using or with its lyophilizing.The pH of compound formulation is generally at 3-11, more preferably 5-9, and 7-8 most preferably.Be appreciated that the application of some causes forming pharmaceutical salts in above-mentioned excipient, carrier or the stabilizing agent.

The therapeutic dose of The compounds of this invention can change according to the administering mode of the concrete application of for example treating, chemical compound, patient's health and situation and the different of judgement of opening according to the clinicist of prescription.Ratio or the concentration of The compounds of this invention in pharmaceutical composition can comprise dosage, chemical characteristic (for example hydrophobicity) and route of administration according to many factors vary.For example, The compounds of this invention can be mixed with the physiological buffer aqueous solution that contains the about 10%w/v chemical compound of the 0.1-that has an appointment so that carry out parenterai administration.Some typical doses scope is about the about 1g/kg body weight/day of 1 μ g/kg-.In certain embodiments, this dosage range is about the about 100mg/kg body weight/day of 0.01mg/kg-.This dosage may depend on such as the type of disease or obstacle to be formed and the such variable of route of administration with development degree, concrete patient's general health, selected relative biological effect, excipient.Can be according to the dose-response curve extrapolation effective dose that derives from external or animal model test macro.

Chemical compound of the present invention can also be prepared with one or more extra active components, described extra active component can comprise any pharmaceutically active agents, such as antibody, immunosuppressant, anti-inflammatory agent, chemotherapeutics, lipid lowerers, high density lipoprotein increasing activating agent, insulin secretagogue or sensitizer, be used for the treatment of the medicine of rheumatoid arthritis etc.

Rheumatoid arthritis (RA) therapeutic scheme

Rheumatoid arthritis (RA) patient who uses disease aromatics modifier (methotrexate, antimalarial, gold, penicillamine, sulfasalazine, dapsone, leflunomide or biological product) to treat in the aggressivity mode can obtain the disease control of variable pitch, comprises fully and disappearing.These clinical responses are relevant with the standardization scoring improvement of disease activity, particularly comprise following ACR standard: the laboratory measurement value (CRP and ESR) of the quantity in pain, function, tenderness joint, the quantity of swollen joint, patient's the overall evaluation, clinicist's the overall evaluation, inflammation and the radiology evaluation of articulation structure damage.The medicine (DMARDs) of present adjusting disease need continue medication so that keep best helpfulness.It is impaired relevant with host defense with tangible toxicity to give these activating agents for a long time.In addition, the patient becomes usually and is difficult to specific therapy for treating and needs alternative.Owing to these reasons, can allow new effective therapy of standard DMARDs cancellation may be important development clinically.

Can the clinical degeneration of disease and the patient who following therapy is had significant reaction have been realized with inhibition CCR2 expression and/or the treatment of active material: anti-TNF therapy (infliximab, Embrel, adalimumab), anti--IL-1 therapy (kinaret) or other are regulated the antirheumatic (DMARDs) of disease, include but not limited to methotrexate, Cyclosporin A, gold salt, antimalarial, penicillamine or leflunomide, described inhibition CCR2 expresses and/or active material comprises: nucleic acid (for example antisense or siRNA molecule) for example, protein (for example anti--CCR2 antibody), micromolecular inhibitor (example is chemical compound and other inhibitors of chemokine receptors as known in the art as disclosed herein).

In certain embodiments, suppressing CCR2 expression and/or active material is micromolecule CCR2 inhibitor (or antagonist).Can give CCR2 antagonist with oral dose q.d. or the b.i.d that is no more than about 500mgs every day.The patient can withdraw from from its present therapy dosage and maybe can reduce this dosage and can be maintained when using the CCR2 treatment.Use CCR2 antagonist therapy present with it to unite the patient is treated, after this about 2 days of for example about 1-stops or reducing the dosage of DMARD and continue to use the CCR2 antagonist for treating.

The advantage of using the CCR2 antagonist to replace traditional DMARDS has a lot.There is the side effect of serious restriction cumulative dose in traditional DMARDs, and modal is damage and immunosuppressive action to liver.Estimate that the CCR2 antagonism has the long-term safety and the not similar immunosuppressant susceptibility relevant with traditional DMARDs of improvement.In addition, the half-life of biological product was generally several days or a few week, and this is a problem when the research untoward reaction.But the order of magnitude that the half-life of estimating the CCR2 antagonist of oral biological utilisation is about hour is compared with biological preparation thus, and the risk that continues the contact medicine after untoward reaction is very small.In addition, generally give present biological preparation (infliximab, Embrel, adalimumab, kinaret), thereby require the doctor to carry out administration or patient's self-injection by i.v. or s.c..This can produce the probability of infusion reaction or injection site reaction.Use the CCR2 antagonist of oral administration can avoid these problems.

Diabetes and insulin resistant therapeutic scheme

Type 2 diabetes mellitus is the one of the main reasons of M ﹠ M in the Western society.In most of patient, this disease is characterised in that pancreas beta cell malfunction, and is attended by the insulin resistant in liver and the surrounding tissue.Based on the basic mechanism of disease association, can utilize the oral therapy for treating type 2 diabetes mellitus of two kinds of general types: insulin secretagogue (sulfonylurea, such as glibenclamide) and insulin sensitiser thing (metformin and thiazolidinediones are such as rosiglitazone).The verified conjoint therapy that is oriented to these two kinds of mechanism can be controlled this sick metabolic deficiency and show the demand that can improve exogenous insulin administration in many cases, yet, insulin resistant develops usually in time, causes the demand that extra insulin is replenished.In addition, the verified prediabetes state that is called metabolism syndrome is characterised in that glucose tolerance reduces, and is particularly relevant with obesity.Insulin resistant is from taking place in most of patient that type 2 diabetes mellitus takes place, and prevents that when these patients can not keep again the glucose homeostasis from hyperglycemia taking place when lacking necessary hyperinsulinemia degree.The outbreak high predicted of insulin resistant composition seizure of disease and increase relevant with the risk that type 2 diabetes mellitus, hypertension and coronary heart disease take place.

Glucose tolerance reduces and one of the strongest correlative factor that develops into type 2 diabetes mellitus from the insulin resistant state is to exist central obesity.The most of patient who suffers from type 2 diabetes mellitus is for obese type and fat own relevant with insulin resistant.Obviously central obesity is the major risk factors that causes the insulin resistant of type 2 diabetes mellitus, and prompting has impelled the generation of insulin resistant and developed into disease from the signal of interior fat.Except that excretory rho factor, obesity is gone back the inducing cell inflammatory reaction, and wherein the macrophage of derived from bone marrow is accumulated in the fat depot, becomes the fatty tissue macrophage.The ratio that the fatty tissue macrophage is accumulated in the fatty tissue is proportional with obesity.But tissue infiltration's macrophage is the source of many inflammatory cytokines of the insulin resistant in the verified induced lipolysis cell.

The MCP-1 that fatty tissue produces is with fat proportional, points out its activity of carrying out the signal conduction by CCR2 also may accumulate in fatty tissue at macrophage and plays an important role.Whether still do not understand MCP-1/CCR2 and interact and directly to cause monocyte recruitement to fatty tissue, whether whether people's macrophage raised to the minimizing of fatty tissue and directly caused short scorching molecule to produce reducing producing directly relevant with insulin resistant with short scorching molecule.

Can show insulin resistant with inhibition CCR2 expression and/or the treatment of active material, promptly both can be for prediabetes (orthoglycemic) also can be the patient of diabetes (hyperglycemic), described inhibition CCR2 expresses and/or active material comprises: for example nucleic acid (for example antisense or siRNA molecule), protein (for example anti--CCR2 antibody), micromolecular inhibitor (example is chemical compound and other inhibitors of chemokine receptors as known in the art as disclosed herein).In certain embodiments, suppressing CCR2 expression and/or active material is micromolecule CCR2 inhibitor (or antagonist).Can give CCR2 antagonist with oral dose q.d. or the b.i.d that is no more than about 500mgs every day.The patient can withdraw from from its present therapy dosage and maybe can reduce this dosage and can be maintained when using the CCR2 treatment.Perhaps, the CCR2 antagonist for treating can be used for replenishing its present therapy so that improve its effect or prevent to develop into further insulin resistant.

Advantage with the replacement of CCR2 antagonist or additional traditional activating agent is a lot.For example, this class activating agent can be used to prevent prediabetes insulin resistant state to develop into diabetic disease states.This class activating agent can reduce or replace using the demand of subsidiary virose insulin sensitiser thing.This class activating agent can also reduce the time limit when needing the demand that exogenous insulin replenishes or prolonging needs to arrive the supplemented with exogenous insulin.

The treatment of atherosclerosis scheme

Atherosclerosis is to be characterised in that fatty material is deposited on the disease on the arterial wall.Speckle comprises the deposit of other material of this class fatty material, cholesterol, cellular waste, calcium and formation tremulous pulse liner.Speckle can grow to the blood flow that is enough to obviously to reduce by tremulous pulse largely.Yet, become unstable and when breaking, more tangible infringement generation comprising.Disruptive speckle produces the clot that can form blocking blood flow or blocking-up or be delivered to the health other parts.If clottage supply with the blood vessel of heart, it can cause heart attack so.If clot has been blocked the blood vessel of supplying with brain, it can cause apoplexy so.Atherosclerosis is a kind of disease of complexity slowly, it generally since the childhood period and when the people grows to person in middle and old age, develop usually.

High-level cholesterol in the blood is the main hazard factor of coronary heart disease.Based on the cholesterol of mainly forming as speckle, by reducing the circulation cholesterol or controlling the development that speckle forms by the high density lipoprotein (HDL) that cholesterol is carried in rising.For example, can be by using or suppressing circulation cholesterol synthesizing in liver to reduce the circulation cholesterol by reducing from food, to replenish.This class medicine that works by these mechanism can comprise the medicine that is used to reduce elevated cholesterol: bile acid absorbent, lipoprotein synthetic inhibitor, cholesterol synthetic inhibitor and Carboxymethylcellulose (fibric acid) derivant.In addition, the circulation HDL that can raise by the nicotinic acid that gives probucol (probuchol) for example or high dose.The therapy that has confirmed to solve number of mechanisms can be slowed down disease progression and be developed into plaque rupture.

Atherosclerosis generally is attended by the cellular inflammation reaction, and wherein the macrophage of derived from bone marrow is accumulated in the fatty streaks along blood vessel wall, thereby becomes foam cell.Foam cell is the verified source of inducing many inflammatory cytokines of plaque progression and can promote the enzyme of speckle loss of stability.Atherosclerosis is organized and is also produced MCP-1, thereby points out it also may play an important role in accumulating as the macrophage of the foam cell in speckle by the activity that CCR2 carries out the signal conduction.Confirmed CCR2-/-mice obviously reducing as the macrophage in the resultant fatty streaks of the hereditary change in high fat diet or the lipid metabolism.

Can be with suppressing that CCR2 expresses and/or the treatment of active material shows the circulation CRP of high circulation cholesterol, low HDL or rising or by imaging alleged occurrence blood vessel wall speckle or there is the atherosclerotic patient of other evidence arbitrarily, described inhibition CCR2 expresses and/or active material comprises: for example nucleic acid (for example antisense or siRNA molecule), protein (for example anti--CCR2 antibody), micromolecular inhibitor (example is chemical compound and other inhibitors of chemokine receptors as known in the art as disclosed herein).In certain embodiments, suppressing CCR2 expression and/or active material is micromolecule CCR2 inhibitor (or antagonist), such as chemical compound of the present invention.Can give CCR2 antagonist with oral dose q.d. or the b.i.d that is no more than about 500mgs every day.The patient can withdraw from from its present therapy dosage and maybe can reduce this dosage and can be maintained when using the CCR2 antagonist for treating.Perhaps, the CCR2 antagonist for treating can be used for replenishing its present therapy so that for example improve it at prevention plaque progression, the stable speckle that has formed or induce effect aspect the ratio degeneration.

Advantage with the replacement of CCR2 antagonist or additional traditional activating agent is a lot.For example, this class activating agent can be used to prevent the unstability stage that plaque progression becomes to have the risk relevant with plaque rupture.This class activating agent can reduce or replace the medicine that uses subsidiary virose cholesterol regulating or the demand of HDL rising medicine, includes but not limited to rubescent, hepatic injury and muscle injury, such as myopathy.This class activating agent can also reduce the needs operation is carried out this operation or used anticoagulant to limit the time limit of the infringement that produces because of potential plaque rupture up to needs up to needs with demand or the prolongation of opening blood vessel wall.

The chemical compound of labelling and test method

Another aspect of the present invention relates to fluorescent dye, spin labeling, heavy metal or radiolabeled compound of Formula I, they not only are used for imaging, and be used for external and the interior mensuration of body, so that in the tissue sample, comprise people's chemokine receptors location and quantitatively, and be used for by labelled compound in conjunction with suppressing to identify chemokine receptor ligands.Therefore, the present invention includes the chemokine receptors that contains this class labelled compound measures.

The present invention further comprises the chemical compound of isotope-labeled general formula I." isotope " or " radiolabeled " chemical compound is a chemical compound of the present invention, and wherein one or more atoms are had, and atomic weight or atomic number are different from (promptly naturally occurring) atomic weight of general practical measurement or the atom of atomic number is replaced or replacement.The suitable radionuclide that can mix The compounds of this invention includes, but are not limited to ²H (also being written as D, deuterium), ³H (also being written as T, tritium), ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ¹⁸F, ³⁵S, ³⁶Cl, ⁸²Br, ⁷⁵Br, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I, ¹²⁵I and ¹³¹I.The radionuclide that can mix the radiolabeled chemical compound of the present invention depends on the concrete application of this radiolabeled chemical compound.For example, with regard to external chemokine receptors labelling and competitive trials, mix ³H, ¹⁴C, ⁸²Br, ¹²⁵I, ¹³¹I, ³⁵S or chemical compound generally useful.With regard to radiological imaging is used, ¹¹C, ¹⁸F, ¹²⁵I, ¹²³I, ¹²⁴I, ¹³¹I, ⁷⁵Br, ⁷⁶Br or ⁷⁷Br is generally useful.

Be appreciated that " radiolabeled " or " chemical compound of labelling " is for mixing the chemical compound of at least a radionuclide.In certain embodiments, radionuclide is selected from ³H, ¹⁴C, ¹²⁵I, ³⁵S and ⁸²The group that Br forms.

The synthetic method that radiosiotope is mixed with organic compounds be applied to chemical compound of the present invention and for this area well-known.Radiolabeled chemical compound of the present invention can be used to identify/screening test of assessing compound.Generally speaking, can estimate binding ability newly synthetic or compounds identified (being testing compound) minimizing radiolabeled chemical compound of the present invention and chemokine receptors.Therefore, testing compound is directly related with its binding affinity with the ability of the competitive binding chemotactic factor receptor of radiolabeled chemical compound.

Test kit

The present invention also comprises the pharmaceutical kit that for example is used for the treatment of or prevents chemotactic factor-relevant disease, and it comprises one or more containers that contain the pharmaceutical composition that comprises the compound of Formula I for the treatment of effective dose.If desired, this class test kit may further include one or more in the various common drug test kit compositions so, such as, for example contain the container of one or more pharmaceutically acceptable carriers, extra container etc., just as apparent to those skilled in the art.In this test kit, can also comprise consumption, the administration guide of the composition that expression gives and/or mix described composition guide can be the technical instruction of inset or label.

By specific embodiment the present invention is described more specifically.It is for task of explanation that the following example is provided, but also limits the present invention never in any form.Those skilled in the art are easy to discern various nonessential parameters, can change or revise them and can produce identical effect basically.

Embodiment

Embodiment 1

N-[(1R, 3S)-3-isopropyl-3-({ 4-[3-(trifluoromethyl) phenyl] piperazine-1-yl } carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A-1

4,4-dimethoxy tetrahydrochysene-2H-pyrans-3-alcohol

To at 0 ℃ of following 4-methoxy ethyl-3,6-dihydro-2H-pyrans (5.00g, 43.8mmol) drip in the solution in methanol (100mL) between-chlorine benzylhydroperoxide (15.1g, 87.6mmol) solution in methanol (15mL).After stirring 5 hours, remove methanol and white residue is dissolved in dichloromethane (300mL) in vacuum.In this solution, add K ₂CO ₃Filter with gained solution stirring 1 hour and by celite.Evaporated filtrate and obtain required product in a vacuum not purifiedly is used for next step reaction with it.

Steps A-2

3,4,4-trimethoxy tetrahydrochysene-2H-pyrans

To at 0 ℃ down 4,4-dimethoxy tetrahydrochysene-2H-pyrans-3-alcohol (6.00g, 37.0mmol) add in the solution in THF (100mL) sodium hydride (1.48g, 37.0mmol).After stirring 1 hour under 0 ℃, and the dropping methyl iodide (4.61mL, 74.0mmol).With this reaction system temperature to ambient temperature and use NH ₄The Cl quencher.With ether product is extracted 3 times.Use Na ₂SO ₄The dry extract that merges and concentrated.Obtain required product by flash chromatography on silica gel method purification (10% ether-60% ether/hexane). ¹H?NMR(CDCl ₃)δ4.05-3.95(1H，m)，3.80-3.70(1H，m)，3.60-3.50(3H，m)，3.50(3H，s)，3.30(3H，s)，3.10(3H，s)，2.00-1.70(2H，m)。

Steps A-3

3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone

To 3,4, (4g is 20mmol) at THF/H for 4-trimethoxy tetrahydrochysene-2H-pyrans ₂Add dense HCl (6mL) in the solution among the O (60mL/10mL).After stirring 1 hour, remove THF in a vacuum.(3 * 100mL) extract aqueous solution with ether.Dry extract and concentrate in a vacuum and obtain required product. ¹H?NMR(CDCl ₃)δ4.30-4.10(2H，m)，3.75-3.65(2H，m)，3.60-3.50(1H，m)，3.50(3H，s)，2.70-2.50(2H，m)。

Step B-1

(1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino] ring penta-2-alkene-1-methyl formate

To (1R, 4S)-4-[(tertbutyloxycarbonyl) amino] ring penta-2-alkene-1-formic acid (10.0g, 44mmol) add in the solution in DMF (25mL) potassium carbonate (6.33g, 45.8mmol), add subsequently iodomethane (4.0mL, 64mmol).After at room temperature stirring is spent the night, dilute this reactant mixture with EtOAc.Water is with this solution washing 4 times and with salt water washing 1 time, dry (MgSO ₄) and concentrate.In high vacuum, the residue dried overnight is obtained title compound (11g, 99%).C ₁₂H ₁₉NO ₄The MS value of calculation: (M+H) ⁺242; Measured value 142.1 (M-Boc+H) ⁺ ¹H?NMR(CDCl ₃)δ5.86(m，2H)，4.90(m，1H)，4.80(m，1H)，3.72(s，3H)，3.50(m，1H)，2.51(m，1H)，1.86(m，1H)，1.42(s，9H)。

Step B-2

(1S, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-isopropyl ring penta-2-alkene-1-methyl formate

In 10 minutes in the solution (202mL) of-78 ℃ of following 1.00M hexamethyl two silicon lithium nitrides in THF, adding (1R, 4S)-and the 4-[(tertbutyloxycarbonyl) amino] ring penta-2-alkene-1-methyl formate (22.10g, 91.59mmol) solution in THF (36.2mL).This solution was stirred 30 minutes down at-78 ℃, after this add for 1 time isopropyl iodide (10.0mL, 100mmol).Then this mixture is gone in the cold closet of reading under-24 ℃ and keep spending the night.Make the reaction quencher and gained solution is extracted 3 times with aqueous ammonium chloride solution with ether.With dried over sodium sulfate ether layer and evaporation in a vacuum.By silicon dioxide purified by flash chromatography residue, obtain title compound (20.2g) with 10% ethyl acetate/hexane eluting.C ₁₅H ₂₅NO ₄The MS value of calculation: (M+H) ⁺284; Measured value 184.2 (M-Boc+H) ⁺

Step B-3

(1S, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-isopropyl ring penta-2-alkene-1-formic acid

To (1S, 4S)-and the 4-[(tertbutyloxycarbonyl) amino]-1-isopropyl ring penta-2-alkene-1-methyl formate (18.42g, 65mmol) add in the solution in THF (500mL), methanol (500mL) and water (100mL) lithium hydroxide monohydrate (5.00g, 119mmol).This mixture heated is spent the night to refluxing.After 18 hours, TLC shows the raw material of denier.Remove organic solvent in a vacuum and extract water layer so that remove unreacted raw material with ether (200mL).With dense HCl water layer is acidified to pH=4, cools off with ice bath simultaneously.With dichloromethane gained solution is extracted 3 times.Use MgSO ₄Dry extract and be concentrated into and obtain solid (17g).This solid is dissolved in hot ethyl acetate (22mL) and hexane (550mL) is joined in this solution.This solution is slowly cooled to room temperature, after this put into reading at-22 to-24 ℃ cold closet.After 2 days, take out crystallization and concentrated liquid and obtain required product in a vacuum, be white solid foam (9.78g, 56%).C ₁₄H ₂₃NO ₄The MS value of calculation: (M+H) ⁺270; Measured value 170.1 (M-Boc+H) ⁺

Step B-4

(1S, 3R)-the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid

To (1S, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-(9.78g 36.3mmol) adds 10% palladium/carbon (550mg) to 1-isopropyl ring penta-2-alkene-1-formic acid in the solution in ethanol (250mL).The jolting in the hydrogen environment under the 55psi of this mixture is spent the night and filter by celite.Evaporated filtrate and obtain title compound (9.45g, 96%) in a vacuum.C ₁₄H ₂₅NO ₄The MS value of calculation: (M+H) ⁺272; Measured value 172.1 (M-Boc+H) ⁺

Step C

[(1R, 3S)-3-(4-[3-(trifluoromethyl) phenyl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate

With (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (0.10g, 0.37mmol), the N-[3-trifluoromethyl] phenylpiperazine (85mg, 0.37mmol), triethylamine (0.10mL, 0.74mmol) and benzotriazole-(0.16g 0.37mmol) mixes in dichloromethane (5mL) 1-base oxygen base three (dimethylamino)-phosphorus hexafluorophosphates and stirring is at room temperature spent the night.Dilute this reactant mixture and use saturated NaHCO with EtOAc ₃Washing.With EtOAc water layer is extracted 3 times.Dry organic layer (the MgSO that merges ₄), concentrate and carry out flash chromatography (50%EtOAc/ hexane-EA) obtains required product (86mg, 53%).C ₂₅H ₃₆F ₃N ₃O ₃The MS value of calculation: (M+H) 484.3; Measured value 384.2 (M+H-Boc).

Step D

(1R, 3S)-3-(4-[3-(trifluoromethyl) phenyl] piperazine-1-base carbonyl) Aminocyclopentane two (trifluoroacetate)

At room temperature be used in trifluoroacetic acid in the dichloromethane (3mL) (3mL, 0.04mol) will [(1R, 3S)-3-(4-[3-(trifluoromethyl) phenyl]-piperazine-1-base carbonyl)-cyclopenta]-(82mg 0.18mmol) handled 1 hour t-butyl carbamate.Concentrate this mixture and obtain required product (98mg, 93%).C ₂₀H ₂₈F ₃N ₃The MS value of calculation of O: (M+H) 384.3; Measured value 384.2.

Step e

N-[(1R, 3S)-3-isopropyl-3-(4-[3-(trifluoromethyl) phenyl] and piperazine-1-yl } carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-3-isopropyl-3-(4-[3-(trifluoromethyl) phenyl] and piperazine-1-yl } carbonyl) two (the trifluoroacetate) (140mg of Aminocyclopentane, 0.23mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (90mg, 0.69mmol) and triethylamine (0.096mL, 0.69mmol) add in the solution in dichloromethane (10mL) sodium triacetoxy borohydride (97mg, 0.46mmol).After at room temperature stirring is spent the night, dilute this reactant mixture and use saturated Na with EtOAc ₂CO ₃Washing.With EtOAc water layer is extracted 3 times.Dry organic layer (the MgSO that merges ₄), concentrate and use silica gel purification, use EtOAc-1%Et ₃N/EtOAc eluting and obtain the required product of 105mg (92%).Obtain two kinds of main isomers by chirality HPLC separated product.With regard to two kinds of isomers, C ₂₆H ₃₈F ₃N ₃O ₃The MS value of calculation: (M+H) 498; Measured value 498.2.

Embodiment 2

3-ethyoxyl-N-[(1R, 3S)-3-isopropyl-3-({ 4-[3-(trifluoromethyl) phenyl] piperazine-1-yl } carbonyl) cyclopenta] preparation of tetrahydrochysene-2H-pyrans-4-amine

Steps A-1

3-ethyoxyl-4,4-dimethoxy tetrahydrochysene-2H-pyrans

To refrigerative 4 in ice bath, (2.0g, (0.60g 15mmol) and with the gained slurry stirred 1 hour 4-dimethoxy tetrahydrochysene-2H-pyrans-3-alcohol 12mmol) slowly to add sodium hydride in the solution in THF (20mL).(1.5mL at room temperature stirs 19mmol) and with this mixture and to spend the night to add iodoethane.Add more sodium hydride (0.6g) and iodoethane (3mL) and continue again to stir and spend the night.Water makes the reaction quencher.Gained solution is extracted 2 times and with ether extraction 2 times with EtOAc.The dry extract that merges concentrates and obtains the required product of 2.1g (90%) with silica gel purification (20%EtOAc/ hexane). ¹H?NMR(CDCl ₃)δ3.32-4.00(7H，m)，3.30(3H，s)，3.20(3H，s)，2.05-1.70(2H，m)，1.25-1.22(3H，m)。

Steps A-2

3-ethyoxyl tetrahydrochysene-4H-pyrans-4-ketone

To 3-ethyoxyl-4, (2.1g 11mmol) adds dense HCl (6mL) to 4-dimethoxy tetrahydrochysene-2H-pyrans in the solution in THF/ water (50mL/10mL).After at room temperature stirring 1 hour, dilute this mixture with EtOAc.Separate organic layer and water layer is extracted 5 times with ether.Use MgSO ₄The dry organic layer that merges filters and is concentrated into and obtains the required product of 1.55g (97%). ¹H?NMR(CDCl ₃)δ4.30-3.50(7H，m)，2.62-2.57(2H，m)，1.30-1.20(3H，t，J＝5Hz)。

Step B

3-ethyoxyl-N-[(1R, 3S)-3-isopropyl-3-(4-[3-(trifluoromethyl) phenyl] and piperazine-1-yl } carbonyl) cyclopenta] tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-3-isopropyl-3-(4-[3-(trifluoromethyl) phenyl] and piperazine-1-yl } carbonyl) Aminocyclopentane dihydrochloride (200mg, 0.438mmol), 3-ethyoxyl tetrahydrochysene-4H-pyrans-4-ketone (130mg, 0.88mmol) and triethylamine (0.18mL, 1.3mmol) add in the solution in dichloromethane (10mL) sodium triacetoxy borohydride (180mg, 0.88mmol).After at room temperature stirring is spent the night, dilute this reactant mixture and use saturated Na with EtOAc ₂CO ₃Washing.With EtOAc water layer is extracted 3 times.Dry organic layer (the MgSO that merges ₄), concentrate and with silica gel purification (EtOAc-1%Et ₃N/EtOAc-5%Et ₃N/EtOAc) obtain the 230mg product.Obtain two kinds of main isomers by chirality HPLC separated product: isomer 1 (110mg) and isomer 2 (77mg).C ₂₇H ₄₀F ₃N ₃O ₃The MS value of calculation: (M+H) 512; Measured value 512.2.

Embodiment 3

N-[(1R, 3S)-3-isopropyl-3-({ 4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-yl } carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A

1-[4-(trifluoromethyl) pyridine-2-yl] piperazine

With 2-chloro-4-(trifluoromethyl) pyridine (2.0g, 11mmol), piperazine (3g, 30mmol) and triethylamine (3.1mL, 22mmol) solution in DMF (10mL) is 100 ℃ of following heated overnight and concentrate in a vacuum.By silica gel column chromatography purification residue (EtOAc-EtOAc/MeOH/Et ₃N=9/1/0.5) obtain 1.09g (43%) pure products.C ₁₀H ₁₂F ₃N ₃The MS value of calculation: (M+H) 232; Measured value 232.1.

Step B

[(1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] and piperazine-1-yl } carbonyl) cyclopenta] t-butyl carbamate

To 1-[4-(trifluoromethyl) pyridine-2-yl] piperazine (145mg, 0.627mmol), (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (140mg, 0.52mmol) add benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (253mg in the solution in dichloromethane (10mL), 0.572mmol), add subsequently triethylamine (0.156mL, 1.12mmol).After stirring is spent the night, dilute this reactant mixture and use saturated NaHCO with EtOAc ₃Washing.With EtOAc water layer is extracted 3 times.Dry organic layer (the MgSO that merges ₄), concentrate and obtain the required product of 0.15g by flash chromatography on silica gel method purification (20% EtOAc/ hexane-40% EtOAc/ hexane).C ₂₄H ₃₅F ₃N ₄O ₃The MS value of calculation: (M+H) 485; Measured value 385.2 (M-Boc+H).

Step C

(1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] and piperazine-1-yl } carbonyl) Aminocyclopentane

At room temperature use 4.0M HCl 1, solution (10mL) in the 4-two  alkane is with [(1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] and piperazine-1-yl } carbonyl) cyclopenta] t-butyl carbamate (150mg, 0.31mmol) handled 1 hour and under reduced pressure be concentrated into and obtain product, it not purifiedly is used for next step.C ₁₉H ₂₇F ₃N ₄The MS value of calculation of O: (M+H) 385; Measured value 385.2.

Step D

N-[(1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] and piperazine-1-yl } carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] and piperazine-1-yl } carbonyl) Aminocyclopentane dihydrochloride (140mg, 0.31mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (80mg, 0.61mmol) and triethylamine (0.21mL, 1.5mmol) add in the solution in dichloromethane (10mL) sodium triacetoxy borohydride (190mg, 0.92mmol).After stirring is spent the night, dilute this reactant mixture and use saturated Na with EtOAc ₂CO ₃Washing.With EtOAc water layer is extracted 3 times.Dry organic layer (the MgSO that merges ₄), concentrate and by flash chromatography on silica gel method purification (EtOAc-1%Et ₃N/EtOAc-5%Et ₃N/EtOAc) obtain the 101mg product.Obtain isomer 1 (55mg) and isomer 2 (37mg) by the further separated product of chirality HPLC.With regard to two kinds of isomers, C ₂₅H ₃₇F ₃N ₄O ₃LCMS value of calculation (M+1) 499; Measured value 499.2.

Embodiment 4

N-[(1R, 3S)-3-isopropyl-3-({ 4-[5-(trifluoromethyl) pyridin-3-yl] piperazine-1-yl } carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A-1

3-bromo-5-iodine pyridine

To 3, (48g 200mmol) adds the solution (80mL) of 2M isopropyl-magnesium chloride in THF to the 5-dibromo pyridine in the solution in THF (200mL).After at room temperature stirring 2 hours, this solution is cooled to-78 ℃.To wherein adding pre-cooled iodine (51g, 200mmol) solution in THF (100mL).Dilute this mixture and with saturated ammonium chloride solution, 2M hypo solution and salt water washing with ether.Use MgSO ₄The dry organic layer that merges filters and concentrates.Crystallization from ethanol and obtain the required product of 33.5g (58%). ¹H?NMR(CDCl ₃)δ8.75(1H，s)，8.60(1H，s)，8.20(1H，s)。

Steps A-2

4-(5-bromopyridine-3-yl) piperazine-1-t-butyl formate

Will the invisible spectro 3-bromo-5-iodine pyridine of sealing (13.0g, 45.8mmol), piperazine-1-t-butyl formate (8.53g, 45.8mmol), Hydro-Giene (Water Science). (I) (0.871g, 4.57mmol), K ₃PO ₄(19.46g, 91.68mmol), 1 (5.1mL, 91mmol) in the oil bath of solution under 80 ℃ in isopropyl alcohol (80mL) heating 2 days.After being cooled to room temperature, filter this reactant mixture by celite.Concentrated filtrate in a vacuum.Residue is dissolved in EtOAc and uses saturated NaHCO ₃Wash this solution, dry (MgSO ₄) and concentrate.Obtain the required product of 5.75g (37%) by flash chromatography on silica gel method purification (20% EtOAc/ hexane-30%EtOAc/ hexane).C ₁₄H ₂₀BrN ₃O ₂The MS value of calculation: (M+H) 343; Measured value 342.0,344.0.

Steps A-3

4-(5-iodine pyridine-3-yl) piperazine-1-t-butyl formate

(2.0g 5.9mmol) adds the solution (5mL) of 2M isopropyl-magnesium chloride in THF in the solution in THF (20mL) to 4-(3-bromophenyl) piperazine-1-t-butyl formate.After at room temperature stirring 2 hours, this solution is cooled to-78 ℃.(3.0g is 12mmol) at THF (2mL) to wherein adding pre-cooled iodine.-78 ℃ stirred 30 minutes down and at room temperature restir dilute this mixture with ethyl acetate after 30 minutes, with saturated ammonium chloride, 2M hypo solution and salt water washing, dry (MgSO ₄) and concentrate.Obtain required product (1.40g) by flash chromatography on silica gel method purification residue (20% EtOAc/ hexane-50%EtOAc/ hexane), purity is 75%.C ₁₅H ₂₁IN ₂O ₂The MS value of calculation: (M+H) 390; Measured value 390.0.

Steps A-4

4-[5-(trifluoromethyl) pyridin-3-yl] piperazine-1-t-butyl formate

Under slow jolting and in the high vacuum with the Hydro-Giene (Water Science). (I) of flame heat in flask (0.49g, 2.6mmol) and potassium fluoride (0.15g, 2.6mmol), up to light green occurring.(0.5g is 1.0mmol) with (trifluoromethyl) trimethyl silane (0.37g, 2.6mmol) solution in DMF (5mL) to add 4-(3-iodophenyl) piperazine-1-t-butyl formate.This brown solution at room temperature stirred spend the night.Add (trifluoromethyl) trimethyl silane (0.37g) again.This mixture 50 ℃ of following heated overnight, is washed with the EtOAc dilution and with saturated ammonium chloride.With EtOAc water layer is extracted 3 times.Dry organic layer (the MgSO that merges ₄), concentrate and obtain the required product of 120mg by flash chromatography on silica gel method purification (20%-40%EtOAc/ hexane).C ₁₅H ₂₀F ₃N ₃O ₂The MS value of calculation: (M+H) 332; Measured value 332.1.

Steps A-5

1-[5-(trifluoromethyl) pyridin-3-yl] piperazine

At room temperature use 4.0M HCl 1, the solution (7mL) in the 4-two  alkane is with 4-[5-(trifluoromethyl) pyridin-3-yl] (0.24g 0.25mmol) handled 1 hour and concentrates piperazine-1-t-butyl formate.Residue is used for next step without being further purified.C ₁₀H ₁₂F ₃N ₃The MS value of calculation: (M+H) 232; Measured value 232.1.

Step B

[(1R, 3S)-3-isopropyl-3-(4-[5-(trifluoromethyl) pyridin-3-yl] and piperazine-1-yl } carbonyl) cyclopenta] t-butyl carbamate

To 1-[5-(trifluoromethyl) pyridin-3-yl] piperazine trihydrochloride salt (0.22g, 0.23mmol), (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (0.18g, 0.66mmol) add benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (0.338g in the solution in dichloromethane (10mL), 0.764mmol), add subsequently triethylamine (0.22mL, 1.6mmol).At room temperature this mixture is stirred and spend the night and dilute with EtOAc.Use saturated NaHCO ₃Wash this solution, dry (MgSO ₄) and concentrate.Obtain the required product of 0.19g (61%) by flash chromatography on silica gel method purification residue (20% EtOAc/ hexane-40%EtOAc/ hexane).C ₂₄H ₃₅F ₃N ₄O ₃The MS value of calculation: (M+H) 485; Measured value 485.2.

Step C

(1R, 3S)-3-isopropyl-3-(4-[5-(trifluoromethyl) pyridin-3-yl] and piperazine-1-yl } carbonyl) Aminocyclopentane

Use 4.0M HCl 1 under the room temperature, solution (5mL) general in the 4-two  alkane [(1R, 3S)-3-isopropyl-3-(4-[5-(trifluoromethyl) pyridin-3-yl] and piperazine-1-yl } carbonyl) cyclopenta] (190mg 0.14mmol) handled 1 hour t-butyl carbamate.Concentrate this mixture and obtain the required product of 35mg by the HPLC purification.C ₁₉H ₂₇F ₃N ₄The MS value of calculation of O; (M+H) 385; Measured value 385.1.

Step D

N-[(1R, 3S)-3-isopropyl-3-(4-[5-(trifluoromethyl) pyridin-3-yl] and piperazine-1-yl } carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-3-isopropyl-3-(4-[5-(trifluoromethyl) pyridin-3-yl] and piperazine-1-yl } carbonyl) Aminocyclopentane three (trifluoroacetate) (25mg, 0.034mmol), 3-methyl tetrahydrochysene-4H-pyrans-4-ketone (13mg, 0.10mmol) and triethylamine (0.024mL, 0.17mmol) add in the solution in dichloromethane (5mL) sodium triacetoxy borohydride (22mg, 0.10mmol).With this mixture at room temperature and N ₂Stir in the environment and spend the night and dilute with EtOAc.Use saturated NaHCO ₃Washing gained solution, dry (MgSO ₄) and concentrate.By flash chromatography on silica gel method purification residue (EtOAc-EtOAc/MeOH/Et ₃N=9: 1: 0.5) obtaining the required product of 14mg, is two kinds of mixture of isomers.Separate two kinds of isomers by chirality HPLC and obtain peak 1 (6.6mg) and peak 2 (4.7mg).With regard to two kinds of isomers, C ₂₅H ₃₇F ₃N ₄O ₃The MS value of calculation: (M+H) 499; Measured value 499.2.

Embodiment 5

N-{ (1R, 3S)-3-isopropyl-3-[(4-phenyl-3,6-dihydropyridine-1 (2H)-yl) carbonyl] cyclopenta }-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A

(1R, 3S)-3-isopropyl-3-[(4-phenyl-3,6-dihydropyridine-1 (2H)-yl) carbonyl] cyclopenta } t-butyl carbamate

In exsiccant flask, will be at N ₂In the environment (1S, 3R)-the 3-[(tertbutyloxycarbonyl) amino]-(200mg, 0.7mmol) with 4-phenyl-1,2,3, (160mg 0.81mmol) is suspended in the dichloromethane (4mL) the 6-tetrahydropyridine 1-isopropyl cyclopentane-carboxylic acid.Add triethylamine (0.22g, 2.2mmol), add subsequently benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (0.36g, 0.81mmol).This reaction system at room temperature stirred spend the night and by adding saturated NaHCO ₃The solution quencher.With dichloromethane gained solution is extracted 3 times.Dry extract (the MgSO that merges ₄), filter and concentrate.By flash chromatography on silica gel method purification (gradient: 0-45% B, in 15 minutes.A bottle=hexane, B bottle=EtOAc) obtain the required product of 234mg (80%).C ₂₅H ₃₆N ₂O ₃The MS value of calculation: (M+H) 413; Measured value 413.2.

Step B

(1R, 3S)-3-isopropyl-3-[(4-phenyl-3,6-dihydropyridine-1 (2H)-yl) carbonyl] Aminocyclopentane

Will (1R, 3S)-3-isopropyl-3-[(4-phenyl-3,6-dihydropyridine-1 (2H)-yl) carbonyl] cyclopenta } (0.23g 0.56mmol) is dissolved in the solution (4mL) of 1.0M HCl in ether to t-butyl carbamate.After at room temperature stirring 2 hours, concentrate this solution to obtaining colorless oil (170mg).C ₂₀H ₂₈N ₂The MS value of calculation of O: (M+H) 313; Measured value 313.2.

Step C

N-{ (1R, 3S)-3-isopropyl-3-[(4-phenyl-3,6-dihydropyridine-1 (2H)-yl) carbonyl]-cyclopenta } tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-3-isopropyl-3-[(4-phenyl-3,6-dihydropyridine-1 (2H)-yl) carbonyl] Aminocyclopentane hydrochlorate (50mg, 0.1mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (56mg, 0.43mmol) and triethylamine (0.070mL, 0.50mmol) add in the solution in dichloromethane (2mL) sodium triacetoxy borohydride (91mg, 0.43mmol).After at room temperature stirring is spent the night, add saturated NaHCO ₃With dichloromethane this solution is extracted 3 times.Dry extract (the MgSO that merges ₄), filter and concentrate.By flash chromatography on silica gel method purification (gradient: 0-15% B, in 15 minutes.A bottle=1% NH ₄OH/3% MeOH/EtOAc, B bottle=1% NH ₄OH/MeOH) obtain required compound.C ₂₆H ₃₈N ₂O ₃The MS value of calculation: (M+H) 427; Measured value 427.3.

Embodiment 6

The preparation of 1-({ (1S, 3R)-1-isopropyl-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino] cyclopenta } carbonyl)-4-Phenylpiperidine-4-alcohol

Steps A

(1R, 3S)-3-[(4-hydroxy-4-phenyl piperidine-1-yl) carbonyl]-3-isopropyl cyclopenta } t-butyl carbamate

To (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-different third Pentamethylene. of 1--formic acid (200mg, 0.7mmol), 4-Phenylpiperidine-4-alcohol (140mg, 0.81mmol) and triethylamine (0.15g, 1.5mmol) add in the solution in dichloromethane (4mL) (benzotriazole-1-base oxygen base) tripyrrole alkane phosphorus hexafluorophosphate (0.42g, 0.81mmol).After at room temperature stirring is spent the night, by adding saturated NaHCO ₃Solution makes the reaction quencher.With dichloromethane gained solution is extracted 3 times.Dry extract (the MgSO that merges ₄), filter and concentrate.This crude product is used for next step without being further purified.C ₂₅H ₃₈N ₂O ₄The MS value of calculation: (M+H) 431; Measured value 431.2.

Step B

1-{[(1S, 3R)-3-amino-1-isopropyl cyclopenta] carbonyl }-4-Phenylpiperidine-4-alcohol

Will (1R, 3S)-3-[(4-hydroxy-4-phenyl piperidine-1-yl) carbonyl]-3-isopropyl-cyclopenta } (0.30g 0.70mmol) is dissolved in the solution (10mL) of 2.0M HCl in ether to t-butyl carbamate.After stirring 3.5 hours, add several MeOH and obtain settled solution.Concentrate this mixture to obtaining grease.Be not used for next step reaction with this crude product is purified.C ₂₀H ₃₀N ₂O ₂The MS value of calculation: (M+H) 331; Measured value 331.2.

Step C

1-((1S, 3R)-1-isopropyl-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino] cyclopenta } carbonyl)-4-Phenylpiperidine-4-alcohol

To 1-{[(1S, 3R)-and 3-amino-1-isopropyl cyclopenta] carbonyl }-4-Phenylpiperidine-4-alcohol hydrochloride (50mg, 0.1mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (53mg, 0.41mmol) and triethylamine (0.066mL, 0.48mmol) add in the solution in dichloromethane (2mL) sodium triacetoxy borohydride (0.087g, 0.41mmol).After at room temperature stirring is spent the night, use saturated NaHCO ₃Solution makes the reaction quencher.With dichloromethane gained solution is extracted 3 times.Dry extract (the MgSO that merges ₄), filter and concentrate.(0-20% B is in 17 minutes by flash chromatography on silica gel method purification.A bottle=1% NH ₄OH/2% MeOH/EtOAc, B bottle=1% NH ₄OH/MeOH) obtain required product.C ₂₆H ₄₀N ₂O ₄The MS value of calculation: (M+H) 445; Measured value 445.2.

Embodiment 7

1-({ (1S, 3R)-1-isopropyl-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino] cyclopenta } carbonyl)-4-[2-(trifluoromethyl) phenyl] preparation of piperidines-4-alcohol

Steps A-1

4-hydroxyl-4-[2-(trifluoromethyl) phenyl] piperidines-1-t-butyl formate

(1.18g 5.24mmol) drips the solution (3.4mL) of 1.60M n-BuLi in hexane in the solution in THF (20mL) to 1-bromo-2-(trifluoromethyl) benzene that is cooled to-78 ℃.After stirring 40 minutes, add 4-oxo-1-piperidine acid tert-butyl ester (1.0g, 5.0mmol) solution in THF (3mL) and this solution stirred 1 hour down at-78 ℃.Make the reaction quencher with saturated ammonium chloride.With dichloromethane gained solution is extracted 3 times.Dry extraction (the MgSO that merges ₄), filter and be concentrated into and obtain the 0.78g white solid, it not purifiedly is used for next step reaction.C ₁₇H ₂₂F ₃NO ₃The MS value of calculation: (M+H) 346; Measured value 246.0 (M-Boc+1).

Steps A-2

4-[2-(trifluoromethyl) phenyl] piperidines-4-alcohol

With 4-hydroxyl-4-[2-(trifluoromethyl) phenyl] (0.40g 1.0mmol) is dissolved in the solution (5mL) of 2.0M HCl in ether to piperidines-1-t-butyl formate.After at room temperature stirring is spent the night, dilute this solution with ether.Filter white solid and wash-obtain the 170mg pure products with ether.C ₁₂H ₁₄F ₃The MS value of calculation of NO: (M+H) 246; Measured value 246.1.

Step B

[(1R, 3S)-3-(4-hydroxyl-4-[2-(trifluoromethyl) phenyl] and piperidines-1-yl } carbonyl)-3-isopropyl cyclopenta] t-butyl carbamate

To (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (150mg, 0.55mmol), 4-[2-(trifluoromethyl) phenyl] piperidines-4-alcohol hydrochloride (170mg, 0.60mmol) and triethylamine (0.17g, 1.6mmol) add in the solution in dichloromethane (3mL) (benzotriazole-1-base oxygen base) tripyrrole alkane phosphorus hexafluorophosphate (0.31g, 0.60mmol).After stirring 2.5 hours, by adding saturated NaHCO ₃Solution makes the reaction quencher.With dichloromethane gained solution is extracted 3 times.Dry extract (the MgSO that merges ₄), filter and concentrate.Do not carry out next step with crude product is purified.C ₂₆H ₃₇F ₃N ₂O ₄The MS value of calculation: (M+H) 499; Measured value 499.2.

Step C

1-{[(1S, 3R)-3-amino-1-isopropyl cyclopenta] carbonyl }-4-[2-(trifluoromethyl) phenyl] piperidines-4-alcohol

To containing (1R, 3S)-3-({ 4-hydroxyl-4-[2-(trifluoromethyl) phenyl] piperidines-1-yl } carbonyl)-3-isopropyl cyclopenta] t-butyl carbamate (0.27g, add in flask 0.54mmol) 2.00M HCl in ether solution (5mL) and the gained mixture stirred 3.5 hours.Concentrate this solution to obtaining grease, it not purifiedly is used for next step reaction.C ₂₁H ₂₉F ₃N ₂O ₂The MS value of calculation: (M+H) 399; Measured value 399.2.

Step D

1-((1S, 3R)-1-isopropyl-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino] cyclopenta } carbonyl)-4-[2-(trifluoromethyl) phenyl] piperidines-4-alcohol

To 1-{[(1S, 3R)-and 3-amino-1-isopropyl cyclopenta] carbonyl }-4-[2-(trifluoromethyl) phenyl] piperidines-4-alcohol hydrochloride (50mg, 0.1mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (45mg, 0.34mmol) and triethylamine (0.048mL, 0.34mmol) add in the solution in dichloromethane (2mL) sodium triacetoxy borohydride (0.073g, 0.34mmol).After at room temperature stirring is spent the night, add saturated NaHCO ₃Solution.With dichloromethane gained solution is extracted 3 times.Dry extract (the MgSO that merges ₄), filter and concentrate.(0-20% B is in 15 minutes by flash chromatography on silica gel method purification.A bottle=1% NH ₄OH/2%MeOH/EtOAc, B bottle=1% NH ₄OH/MeOH) obtain the required product of 15mg, be grease.C ₂₇H ₃₉F ₃N ₂O ₄The MS value of calculation: (M+H) 513; Measured value 513.2.

Embodiment 8

1-[((1S, 3R)-1-isopropyl-3-{[3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl] amino } cyclopenta) carbonyl]-4-[3-(trifluoromethyl) phenyl] preparation of piperidines-4-alcohol

Be used in embodiment 7 described similar operation steps are prepared title compound.C ₂₇H ₃₉F ₃N ₂O ₄The MS value of calculation: (M+H) 513; Measured value 513.2.

Embodiment 9

1-[((1S, 3R)-1-isopropyl-3-{[3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl] amino } cyclopenta) carbonyl]-4-[4-(trifluoromethyl) phenyl] preparation of piperidines-4-alcohol

Be used in embodiment 21 described similar operation steps are prepared title compound.C ₂₇H ₃₉F ₃N ₂O ₄The MS value of calculation: (M+H) 513; Measured value 513.2.

Embodiment 10

N-((1R, 3S)-3-isopropyl-3-{[4-[2-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] carbonyl } cyclopenta)-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A-1

4-[2-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-t-butyl formate

To refrigerative 4-hydroxyl-4-[2-(trifluoromethyl) phenyl in ice bath] piperidines-1-t-butyl formate (0.75g, 2.2mmol) (0.79mL is 11mmol) and with this mixture temperature to room temperature and stirring spend the night (17 hours) slowly to add thionyl chloride in the solution in pyridine (15mL).Make the reaction quencher with frozen water.With dichloromethane gained solution is extracted 3 times.Dry extract (the MgSO that merges ₄), filter and concentrate.(0-40% B is in 25 minutes by flash chromatography on silica gel method purification.A bottle=hexane, B bottle=EtOAc) obtain the required product of 209g is solid.C ₁₇H ₂₀F ₃NO ₂The MS value of calculation: (M+H) 328; Measured value 228.0 (M-Boc+H).

Steps A-2

4-[2-(trifluoromethyl) phenyl]-1,2,3, the 6-tetrahydropyridine

To 4-[2-(trifluoromethyl) phenyl]-3, (200mg 0.61mmol) adds trifluoroacetic acid (2.5mL) in the solution in dichloromethane (5mL) to 6-dihydropyridine-1 (2H)-t-butyl formate.After at room temperature stirring 45 minutes, with this solution concentration to obtaining grease.

Step B

((1R, 3S)-3-isopropyl-3-{[4-phenyl-2-(trifluoromethyl)-3,6-dihydropyridine-1 (2H)-yl] carbonyl } cyclopenta) t-butyl carbamate

To (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (115mg, 0.424mmol) and 4-[2-(trifluoromethyl) phenyl]-1,2,3,6-tetrahydropyridine trifluoroacetate (152mg, 0.445mmol) add triethylamine (0.21g in the solution in dichloromethane (2mL), 2.1mmol), add subsequently benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (210mg, 0.47mmol).After at room temperature stirring is spent the night, use saturated NaHCO ₃Solution makes the reaction quencher.With dichloromethane gained solution is extracted 3 times.Dry extract (the MgSO that merges ₄), filter and concentrate.(0-50% B is in 15 minutes by flash chromatography on silica gel method purification.A bottle=hexane, B bottle=EtOAc) obtain the required product of 174mg is white solid.C ₂₆H ₃₅F ₃N ₂O ₃The MS value of calculation: (M+H) 481; Measured value 481.1.

Step C

(1R, 3S)-3-isopropyl-3-{[4-[2-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] carbonyl } Aminocyclopentane

Will ((1R, 3S)-3-isopropyl-3-{[4-[2-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] carbonyl } cyclopenta) (0.17g 0.00035mol) is dissolved in the solution (2.2mL) of 2.0M HCl in ether to t-butyl carbamate.After at room temperature stirring 2 hours, this solution concentration to obtaining the required product of 144mg, is clarification grease.C ₂₁H ₂₇F ₃N ₂The MS value of calculation of O: (M+H) 381; Measured value 381.1.

Step D

N-((1R, 3S)-3-isopropyl-3-{[4-[2-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] carbonyl } cyclopenta) tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-and 3-isopropyl-3-{[4-[2-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] carbonyl } Aminocyclopentane hydrochlorate (46mg, 0.11mmol), 3-methoxyl group-tetrahydrochysene-4H-pyrans-4-ketone (56mg, 0.43mmol) and triethylamine (0.054mL, 0.39mmol) add in the solution in dichloromethane (2mL) sodium triacetoxy borohydride (70mg, 0.33mmol).After at room temperature stirring is spent the night, add saturated NaHCO ₃Solution.With dichloromethane gained solution is extracted 3 times.Dry extract (the MgSO that merges ₄), filter and concentrate.By flash chromatography on silica gel method purification (0-20% B, 15 minutes.A bottle=1% NH ₄OH/2%MeOH/EtOAc, B bottle=1% NH ₄OH/MeOH) obtain required product.C ₂₇H ₃₇F ₃N ₂O ₃The MS value of calculation: (M+H) 495; Measured value 495.2.

Embodiment 11

N-((1R, 3S)-3-isopropyl-3-{[4-[3-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] carbonyl } cyclopenta)-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

According to embodiment 10 described similar modes are prepared title compound.C ₂₇H ₃₇F ₃N ₂O ₃MS value of calculation (M+H): 495; Measured value 495.2.

Embodiment 12

3-ethyoxyl-N-((1R, 3S)-3-isopropyl-3-{[4-[3-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] carbonyl } cyclopenta) preparation of tetrahydrochysene-2H-pyrans-4-amine

According to embodiment 10 described similar modes are prepared title compound.C ₂₈H ₃₉F ₃N ₂O ₃The MS value of calculation: (M+H) 509; Measured value 509.1.

Embodiment 13

N-((1R, 3S)-3-isopropyl-3-{[4-(trifluoromethyl)-3 ', 6 '-dihydro-2,4 '-preparation of bipyridyl-1 ' (2 ' H)-yl] carbonyl } cyclopenta)-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A-1

2-bromo-4-(trifluoromethyl) pyridine

(2.70g, 14.9mmol) (3.90mL, 29.6mmol) mixture in propionitrile (15.0mL) heated 22 hours under reflux state with trimethylammonium bromide silane with 2-chloro-4-(trifluoromethyl) pyridine.Product (having volatility) carefully is rotated to be evaporated to obtains the dense condensed light brown suspension w/o of 4.07g (containing propionitrile), be further purified.C ₆H ₃BrF ₃The LC-MS value of calculation (M+H) 226.9 of N; Measured value 225.9/227.8.

Steps A-2

4-hydroxyl-4-[4-(trifluoromethyl) pyridine-2-yl] piperidines-1-t-butyl formate

2-bromo-4-(trifluoromethyl) pyridine (4.0g, 14.2mmol) solution (9.65mL) of adding 1.6M n-BuLi in hexane in the solution in dry methylene chloride (52.7mL) to refrigerative slight muddiness under-78 ℃.After stirring 40 minutes under-78 ℃, drip 4-oxo-1-piperidine acid tert-butyl ester (2.59g, 12.9mmol) solution in dry methylene chloride (10.0mL).This reaction system was descended stirring 1 hour and used NH at-78 ℃ ₄The quencher of Cl aqueous solution.Remove THF by rotary evaporation.With dichloromethane water layer is extracted 3 times.The dry organic layer that merges filters and concentrates.By silica gel column chromatography purification residue (30: the 70EtOAc/ hexane) obtain the required product of 2.63g (59%), be brown oil.C ₁₆H ₂₁F ₃N ₂O ₃The LC-MS value of calculation: (M+H) 347; Measured value 247.0 (M-Boc+1).

Steps A-3

4-(trifluoromethyl)-3 ', 6 '-dihydro-2,4 '-bipyridyl-1 ' (2 ' H)-t-butyl formate

To refrigerative 4-hydroxyl-4-[4-(trifluoromethyl) pyridine-2-yl in ice bath] piperidines-1-t-butyl formate (2.00g, 2.31mmol) slowly add in the solution in pyridine (15.9mL) thionyl chloride (0.84mL, 12mmol).With this mixture temperature to room temperature and stirring spend the night (17 hours).With the brown reactant mixture of frozen water quencher and with dichloromethane extraction 3 times.Dry extract (the MgSO that merges ₄), filter and concentrate.(0-10%B is in 25 minutes by flash chromatography on silica gel method purification.A bottle=hexane, B bottle=EtOAc) obtain the required product of 404mg (53%) is light brown oily thing.C ₁₆H ₁₉F ₃N ₂O ₂The LC-MS value of calculation: (M+H) 329; Measured value 273.1 (M-tBu+1).

Steps A-4

4-(trifluoromethyl)-1 ', 2 ', 3 ', 6 '-tetrahydrochysene-2,4 '-bipyridyl

With 4-(trifluoromethyl)-3 ', 6 '-dihydro-2,4 '-bipyridyl-1 ' ((380.0mg 1.157mmol) is dissolved in 4M HCl 1 to 2 ' H)-t-butyl formate, the solution (12.0mL) in the 4-two  alkane and form faint yellow clarification (then become muddy) solution.After at room temperature stirring 1 hour, concentrating this reactant mixture in a vacuum and obtain the 389mg product, is yellow natural gum.C ₁₁H ₁₁F ₃N ₂The LC-MS value of calculation: (M+H) 229; Measured value 229.1.

Step B

((1R, 3S)-3-isopropyl-3-{[4-(trifluoromethyl)-3 ', 6 '-dihydro-2,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } cyclopenta) t-butyl carbamate

To (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (0.288g, 1.06mmol) and 4-(trifluoromethyl)-1 ', 2 ', 3 ', 6 '-tetrahydrochysene-2,4 '-the bipyridyl dihydrochloride (0.320g, 1.06mmol) add in the solution in dry methylene chloride (11.5mL) triethylamine (0.592mL, 4.25mmol), add subsequently benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (0.517g, 1.17mmol).After at room temperature stirring is spent the night, use NaHCO ₃With the brown reactant mixture of salt water washing, dry (MgSO ₄), filter and concentrate.Obtain faint yellow solid product: 265mg (52%) by silica gel column chromatography purification residue (30: 70 EtOAc/ hexanes).C ₂₅H ₃₄F ₃N ₃O ₃The LC-MS value of calculation: (M+H) 482; Measured value 382.2 (M-Boc+1).

Step C

(1R, 3S)-3-isopropyl-3-{[4-(trifluoromethyl)-3 ', 6 '-dihydro-2,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } Aminocyclopentane

With ((1R, 3S)-3-isopropyl-3-{[4-(trifluoromethyl)-3 ', 6 '-dihydro-2,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } cyclopenta) t-butyl carbamate (260.0mg, 0.54mmol) be dissolved in 4M HCl 1, the solution (6mL) in the 4-two  alkane and form faint yellow settled solution.After at room temperature stirring 1 hour, concentrating this reactant mixture in a vacuum and obtain the 300mg product, is two-HCl salt.With this solid of 1M NaOH solution-treated.With dichloromethane free alkali is extracted 3 times.The dry extract that merges filters and concentrates and obtains 194mg (94%) product, is faint yellow natural gum.C ₂₀H ₂₆F ₃N ₃The LC-MS value of calculation (M+H) 382 of O; Measured value 382.1.

Step D

N-((1R, 3S)-3-isopropyl-3-([4-(trifluoromethyl)-3 ', 6 '-dihydro-2,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } cyclopenta)-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

At room temperature and N ₂Use sodium triacetoxy borohydride (85.4mg in the environment, 0.403mmol) processing (1R, 3S)-3-isopropyl-3-{[4-(trifluoromethyl)-3 ', 6 '-dihydro-2,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl Aminocyclopentane (51.2mg, 0.134mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (70mg, 0.40mmol) and triethylamine (0.0374mL, 0.269mmol) at dry methylene chloride (5.0mL, 0.078mol) solution in.Use NaHCO ₃Aqueous solution makes the reaction quencher and dilutes with dichloromethane.Separate organic layer and water layer is extracted 3 times with dichloromethane.Use MgSO ₄The dry extract that merges filters and vapourisation under reduced pressure.Make crude product (90mg) by short silicagel pad (30: 70 MeOH/EtOAc).Concentrated filtrate and separate by chirality HPLC and to obtain two kinds of isomers: first kind of isomer 26.3mg; Second kind of isomer 17.3mg.With regard to two kinds of isomers, C ₂₆H ₃₆F ₃N ₃O ₃The MS value of calculation: (M+H) 496; Measured value 496.2.

Embodiment 14

N-((1R, 3S)-3-isopropyl-3-{[5-(trifluoromethyl)-3 ', 6 '-dihydro-3,4 '-preparation of bipyridyl-1 ' (2 ' H)-yl] carbonyl } cyclopenta)-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A-1

3-nitro-5-(trifluoromethyl) pyridine-2-alcohol

At room temperature (10.0g 61.31mmol) joins in the concentrated sulphuric acid (50.0mL) of stirring with 5-(trifluoromethyl) pyridine-2-alcohol.The gained settled solution put into ice-water bath and slowly add potassium nitrate (12.4g, 123mmol), simultaneously with temperature maintenance at 0 ℃.The gained mixture was heated 4 hours down at 65 ℃, after this be poured on ice and carefully use 50% NaOH (83mL) to handle to pH=8.With EtOAc with this extraction with aqueous solution 3 times.The dry extract that merges filters and is concentrated into and obtains 9.78 (77%) crude products (purity＞90%), is yellow solid.Obtain the 8.40g pure products by grinding with EtOAc to be further purified.C ₆H ₃F ₃N ₂O ₃The LC-MS value of calculation: (M+H) 209; Measured value 209.0.

Steps A-2

2-chloro-3-nitro-5-(trifluoromethyl) pyridine

To phosphoryl chloride phosphorus oxychloride (2.0mL, 21.2mmol) and quinoline (1.30mL adds 3-nitro-5-(trifluoromethyl) pyridine-2-alcohol (4.00g, 18.3mmol) (95% purity) pressed powder in solution 10.8mmol).Gained dark-brown suspension was heated to backflow 4 hours and progressively became very muddy dark-brown solution.After being cooled to 100 ℃, water (11mL) is slowly added to fall in this mixture, further be cooled to room temperature and use Na ₂CO ₃Careful neutralization.With EtOAc gained solution is extracted 3 times.The united extraction thing is used MgSO ₄Drying is filtered and evaporation in a vacuum.Obtain the required product of 2.28g by flash chromatography on silica gel method purification residue (EtOAc/ hexane 30: 70).

Steps A-3

5-(trifluoromethyl) pyridine-3-amine

To at N ₂In 2-chloro-3-nitro-5-(trifluoromethyl) pyridine (1.25g 5.518mmol) adds palladium (1.17g, 1.10mmol) (10% dry weight on wet active carbon) in the solution in methanol (25.0mL).Be placed on this reactant mixture on the Parr instrument and hydrogenation 90 minutes under 50psi.Filter out catalyst by the celite pad.Concentrated filtrate and obtain crude product (1.08g), it is enough pure (confirming＞98% by HPLC), need not to be further purified.C ₆H ₅F ₃N ₂The LC-MS value of calculation: (M+H) 163; Measured value 163.1.

Steps A-4

3-bromo-5-(trifluoromethyl) pyridine

(402mg, 5.83mmol) solution in water (6.8mL) slowly joins 5-(trifluoromethyl) pyridine-3-amine (947mg, 5.55mmol) (48% aqueous solution is 1.57mL) in the suspension at hydrogen bromide with sodium nitrite in ice-water-bath.After stirring 10 minutes, directly but be not the cuprous bromide (I) that lentamente the orange diazonium solution of gained is transferred to stirring (876mg, 6.11mmol) and hydrogen bromide (48% aqueous solution is in mixture 0.38mL).The gained brown mixture was heated 1 hour down at 60 ℃.After being cooled to room temperature, dilute this mixture with dichloromethane, with 50% NaOH (up to pH=11) and water dilution.Use the dichloromethane extraction water layer.Carefully concentrate the organic extract that merges in a vacuum and obtain crude product, need not to be further purified.

Steps A-5

4-hydroxyl-4-[5-(trifluoromethyl) pyridin-3-yl] piperidines-1-t-butyl formate

(2.20g, 30% purity 2.92mmol) add the solution (1.99mL) of 1.6M n-BuLi in hexane in the solution of the slight muddiness in dry methylene chloride (15.0mL) to 3-bromo-5-(trifluoromethyl) pyridine under-78 ℃.After stirring 30 minutes under-78 ℃, drip 4-oxo-1-piperidine acid tert-butyl ester (0.534g, 2.65mmol) solution in dry methylene chloride (3.0mL).This reaction system was descended stirring 1.5 hours and used NH at-78 ℃ ₄The quencher of Cl aqueous solution.With dichloromethane gained solution is extracted 3 times.The dry organic layer that merges filters and concentrates.Obtain the required product of 330mg (25%) by silica gel column chromatography purification residue (50: 50 EtOAc/ hexanes), be yellow oil.C ₁₆H ₂₁F ₃N ₂O ₃The LC-MS value of calculation: (M+H) 347; Measured value 247.1 (M-Boc+1).

Steps A-6

5-(trifluoromethyl)-3 ', 6 '-dihydro-3,4 '-bipyridyl-1 ' (2 ' H)-t-butyl formate

To refrigerative 4-hydroxyl-4-[5-(trifluoromethyl) pyridin-3-yl in ice bath] piperidines-1-t-butyl formate (300mg, 0.433mmol) add in the solution in pyridine (3.00mL) thionyl chloride (0.158mL, 2.16mmol).At room temperature stir the back (16 hours) of spending the night, make brown reactant mixture quencher and with dichloromethane extraction 3 times with frozen water.Dry extract (the MgSO that merges ₄), filter and concentrate.(0-20% B is in 35 minutes by flash chromatography on silica gel method purification.A bottle=hexane, B bottle=EtOAc) obtain the required product of 65mg (46%) is faint yellow oily thing.C ₁₆H ₁₉F ₃N ₂O ₂The LC-MS value of calculation: (M+H) 329; Measured value 329.1.

Steps A-7

5-(trifluoromethyl)-1 ', 2 ', 3 ', 6 '-tetrahydrochysene-3,4 '-bipyridyl

With 5-(trifluoromethyl)-3 ', 6 '-dihydro-3,4 '-bipyridyl-1 ' ((50.0mg 0.152mmol) is dissolved in 4M 1 to 2 ' H)-t-butyl formate, the HCl (2mL) in the 4-two  alkane and form faint yellow clarification (then become muddy) solution.After at room temperature stirring 1 hour, concentrating this reactant mixture in a vacuum and obtain 40.0mg (87%) product, is yellow natural gum.C ₁₁H ₁₁F ₃N ₂The LC-MS value of calculation: (M+H) 229; Measured value 229.0.

Step B

((1R, 3S)-3-isopropyl-3-{[5-(trifluoromethyl)-3 ', 6 '-dihydro-3,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } cyclopenta) t-butyl carbamate

To (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (40.0mg, 0.147mmol) and 5-(trifluoromethyl)-1 ', 2 ', 3 ', 6 '-tetrahydrochysene-3,4 '-the bipyridyl dihydrochloride (44.4mg, 0.147mmol) add in the solution in dry methylene chloride (2.5mL) triethylamine (0.103mL, 0.737mmol), add subsequently benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (71.7mg, 0.162mmol).After at room temperature stirring is spent the night, use NaHCO ₃Aqueous solution makes the reaction quencher.With dichloromethane gained solution is extracted 3 times.The dry extract that merges filters, and concentrates.By silicagel column purification residue (30: 70 EtOAc/ hexanes, gradient elutions to 50 then: 50 EtOAc/ hexanes) obtain lark gel product: 24mg (34%) .C ₂₅H ₃₄F ₃N ₃O ₃The LC-MS value of calculation: (M+H) 482; Measured value 382.0 (M-Boc+1).

Step C

(1R, 3S)-3-isopropyl-3-{[5-(trifluoromethyl)-3 ', 6 '-dihydro-3,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } Aminocyclopentane

With ((1R, 3S)-3-isopropyl-3-{[5-(trifluoromethyl)-3 ', 6 '-dihydro-3,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } cyclopenta) t-butyl carbamate (24.0mg, 0.0498mmol) be dissolved in 4M 1, the HCl (2.0mL) in the 4-two  alkane and form faint yellow settled solution.After at room temperature stirring 1 hour, concentrate this reactant mixture in a vacuum.With 1M NaOH solution-treated residue and with dichloromethane this solution is extracted 3 times.Dry extract filters and concentrates and obtains the 28mg product, is faint yellow solid.C ₂₀H ₂₆F ₃N ₃The LC-MS value of calculation of O: (M+H) 382; Measured value 382.1.

Step D

N-((1R, 3S)-3-isopropyl-3-{[5-(trifluoromethyl)-3 ', 6 '-dihydro-3,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } cyclopenta)-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-3-isopropyl-3-{[5-(trifluoromethyl)-3 ', 6 '-dihydro-3,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } Aminocyclopentane (9.0mg, 0.024mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (12.3mg, 0.0709mmol) and triethylamine (6.58 μ L, 0.0472mmol) add in the solution in dry methylene chloride (2.0mL) sodium triacetoxy borohydride (15.0mg, 0.0708mmol).After at room temperature stirring is spent the night, use NaHCO ₃Aqueous solution makes the reaction quencher and dilutes with dichloromethane.Separate organic layer and water layer is extracted 3 times with dichloromethane.Merge organic layer, use MgSO ₄Drying is filtered and vapourisation under reduced pressure.Obtain the pure products of 1.6mg (14%) by silicagel column purification crude product (30: 70 MeOH/EtOAc).C ₂₆H ₃₆F ₃N ₃O ₃The MS value of calculation: (M+H) 496; Measured value 496.1.

Embodiment 15

N-[(1R, 3S)-3-isopropyl-3-({ 4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-yl } carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A

2-piperazine-1-base-4-(trifluoromethyl) pyrimidine

Sealing in vitro with 2-chloro-4-(trifluoromethyl) pyrimidine (2.0g, 11mmol), piperazine (2.8g, 33mmol) and triethylamine (3.0mL, 22mmol) solution in DMF (10mL) 100 ℃ down stirring spend the night.After removing most of solvent, by silica gel column chromatography purification residue (EtOAc-EtOAc/MeOH/NEt ₃9/1/0.5) and obtain the required product of 1.48g (56%).C ₉H ₁₁F ₃N ₄The MS value of calculation: (M+H) 233; Measured value 233.1.

Step B

[(1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] and piperazine-1-yl } carbonyl) cyclopenta] t-butyl carbamate

To 2-piperazine-1-base-4-(trifluoromethyl) pyrimidine (250mg, 1.08mmol), (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (300mg, 1.1mmol) and triethylamine (0.45mL, 3.2mmol) add in the solution in dichloromethane (10mL) benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (520mg, 1.2mmol).After stirring is spent the night, use saturated NaHCO ₃Make the reaction quencher.With EtOAc gained solution is extracted 3 times.Dry organic layer (the MgSO that merges ₄) and concentrate.Obtain the required product of 290mg by silica gel column chromatography purification (20%-40% EtOAc/ hexane).C ₂₃H ₃₄F ₃N ₅O ₃The MS value of calculation: (M+H) 486; Measured value 386.1 (M-Boc+1).

Step C

(1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] and piperazine-1-yl } carbonyl) Aminocyclopentane

Will [(1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] and piperazine-1-yl } carbonyl) cyclopenta] (290mg 0.60mmol) is dissolved in 4.0MHCl 1 to t-butyl carbamate, the solution (10mL) in the 4-two  alkane.After at room temperature stirring 1 hour, concentrate this mixture to obtaining the required product of 270mg.C ₁₈H ₂₆F ₃N ₅The MS value of calculation of O: (M+H) 386; Measured value 386.1.

Step D

N-[(1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] and piperazine-1-yl } carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] and piperazine-1-yl } carbonyl) Aminocyclopentane dihydrochloride (135.0mg, 0.29mmol), 3-methyl tetrahydrochysene-4H-pyrans-4-ketone (100mg, 0.88mmol) and triethylamine (0.16mL, 1.2mmol) add in the solution in dichloromethane (8mL) sodium triacetoxy borohydride (190mg, 0.88mmol).After at room temperature stirring is spent the night, use saturated NaHCO ₃Make the reaction quencher.With EtOAc gained solution is extracted 3 times.Dry organic layer (the MgSO that merges ₄), concentrate.By flash chromatography on silica gel method purification residue (EtOAc-EtOAc/Et ₃N=10: 0.1) obtaining required product, is two kinds of isomer mixtures.Separate two kinds of isomers by chirality HPLC and obtain isomer 1 (change into tfa salt after 65mg) and isomer 2 (change into tfa salt after 45mg).With regard to two kinds of isomers, C ₂₄H ₃₆F ₃N ₅O ₃The MS value of calculation: (M+1) 500; Measured value 500.1.

Embodiment 16

N-[(1R, 3S)-3-isopropyl-3-({ 4-[6-(trifluoromethyl) pyridine-2-yl] piperazine-1-yl } carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A

1-[6-(trifluoromethyl) pyridine-2-yl] piperazine

The sealing in vitro with 2-chloro-6-(trifluoromethyl) pyridine (1.0g, 5.5mmol), piperazine (1.4g, 16.0mmol) and triethylamine (1.5mL, solution 11.0mmol) mix in DMF (10mL).With this mixture 100 ℃ of following heated overnight.Concentrate this reactant mixture and carry out silica gel chromatography (ethyl acetate-EA/MeOH/Et ₃N=9: 1: 0.5) obtains the required product of 1.05g.C ₁₀H ₁₂F ₃N ₃The MS value of calculation: (M+H) 232.1; Measured value 232.1.

Step B

[(1R, 3S)-3-isopropyl-3-(4-[6-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate

To 1-[6-(trifluoromethyl) pyridine-2-yl] piperazine (249mg, 1.08mmol), (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (300mg, 1.10mmol), triethylamine (0.45mL, 3.2mmol) add in the solution in dichloromethane (10mL) benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (524mg, 1.18mmol).After at room temperature stirring is spent the night, use saturated NaHCO ₃Make the reaction quencher.With EtOAc gained solution is extracted 3 times.Dry organic layer (the MgSO that merges ₄) and concentrate.Obtain the required product of 310mg, C by silica gel column chromatography purification (20% EA/ hexane-40% EA/ hexane) ₂₄H ₃₆F ₃N ₄O ₃The MS value of calculation: (M+H) 485.3; Measured value 485.3.

Step C

(1R, 3S)-3-isopropyl-3-(4-[6-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) the Aminocyclopentane dihydrochloride

Will [(1R, 3S)-3-isopropyl-3-(4-[6-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] (300mg 0.62mmol) is dissolved in 4.0M HCl 1 to t-butyl carbamate, the solution (10mL) in the 4-two  alkane.After at room temperature stirring 1 hour, concentrate this solution and obtain the required product of 260mg.C ₁₉H ₂₇F ₃N ₄The MS value of calculation of O: (M+H) 385.2; Measured value 385.2.

Step D

N-[(1R, 3S)-3-isopropyl-3-(4-[6-(trifluoromethyl) pyridine-2-yl] and piperazine-1-yl } carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-and 3-isopropyl-3-(4-[6-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) Aminocyclopentane dihydrochloride (110mg, 0.24mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (80mg, 0.61mmol) and triethylamine (0.16mL, 1.2mmol) add in the solution in dichloromethane (8mL) sodium triacetoxy borohydride (190mg, 0.88mmol).At room temperature stir spend the night after, this reactant mixture of dilute with water and with ethyl acetate extraction 3 times use dried over sodium sulfate, filtration and concentrated in a vacuum.By the thick residue (ethyl acetate-EtOAc/Et of flash column chromatography purification ₃N=10: 0.1) obtaining the required product of 123mg, is two kinds of mixture of isomers.Separate two kinds of isomers by chirality HPLC and obtain isomer 1 (change into tfa salt after 45mg) and isomer 2 (change into tfa salt after 35mg).With regard to two kinds of isomers, C ₂₅H ₃₇F ₃N ₄O ₃The LCMS value of calculation: (M+H) 498.2; Measured value 498.2.

Embodiment 17

N-[(1R, 3S)-3-isopropyl-3-(4-[6-(trifluoromethyl) pyrimidine-4-yl] piperazine-1-base carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A

4-chloro-6-(trifluoromethyl) pyrimidine

With 6-(trifluoromethyl) pyrimidine-4-alcohol (5.0g, 30.5mmol), phosphoryl chloride phosphorus oxychloride (3.41mL, 36.6mmol) and quinoline (2.16mL, 18.3mmol) solution in toluene (50mL) stirred 5 hours down at 100 ℃.This reaction system of dilute with water and with ethyl acetate extraction 3 times is used dried over sodium sulfate, filters and concentrates in a vacuum.Obtain required product (1.20g, 21.6%) by the thick residue of flash column chromatography purification (10% EtOAc/ hexane). ¹H?NMR(400MHz，CDCl3)：9.21ppm(1H，s)，7.78(1H，s)。

Step B

4-piperazine-1-base-6-(trifluoromethyl) pyrimidine

With 4-chloro-6-(trifluoromethyl) pyrimidine (1.0g, 5.48mmol), piperazine (2.36g, 27.4mmol) and triethylamine (2.29mL, 16.4mmol) solution in DMF (20mL) stirred 5 hours down at 100 ℃.This reaction solution of dilute with water and with ethyl acetate extraction 3 times is used dried over sodium sulfate, filters and concentrates in a vacuum.By the thick residue of flash column chromatography purification (10% MeOH/5% Et ₃N/EtOAc) obtain required product (720mg, 56.6%).C ₉H ₁₂F ₃N ₄The LCMS value of calculation: (M+H) 233.1; Measured value 233.1.

Step C

[(1R, 3S)-3-isopropyl-3-(4-[6-(trifluoromethyl) pyrimidine-4-yl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate

With 4-piperazine-1-base-6-(trifluoromethyl) pyrimidine (1.0g, 4.31mmol), (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (1.75g, 6.46mmol), benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (2.86g, 6.46mmol) and triethylamine (1.20mL, 8.61mmol) solution in dichloromethane (10mL) at room temperature stirs and spends the night.Dilute this reactant mixture with dichloromethane, use the salt water washing, used dried over sodium sulfate, filter and concentrate in a vacuum.Obtain required product (800mg, 38.3%) by the thick residue of flash column chromatography purification.C ₂₃H ₃₅F ₃N ₅O ₃The LCMS value of calculation: (M+H) 486.2; Measured value 486.2.

Step D

To be dissolved in 4M 1, HCl (10mL in the 4-two  alkane, 40mmol) [(1R, 3S)-3-isopropyl-3-(4-[6-(trifluoromethyl) pyrimidine-4-yl] piperazine-1-base carbonyl) cyclopenta] (800mg, solution 1.65mmol) at room temperature stirred 2 hours t-butyl carbamate.Dilute this reactant mixture with dichloromethane, use saturated NaHCO ₃Solution washing is used dried over sodium sulfate, filters and concentrates in a vacuum.Obtain required product (0.6g, 99%) by the thick residue of flash column chromatography purification.C ₁₈H ₂₇F ₃N ₅The LCMS value of calculation of O: (M+H) 386.2; Measured value 386.2.

Step e

N-[(1R, 3S)-3-isopropyl-3-(4-[6-(trifluoromethyl) pyrimidine-4-yl] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-and 3-isopropyl-3-(4-[6-(trifluoromethyl) pyrimidine-4-yl] piperazine-1-base carbonyl) Aminocyclopentane (120mg, 0.30mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (120mg, 0.90mmol) and triethylamine (0.12mL, 0.90mmol) add in the solution in dichloromethane (20mL) sodium triacetoxy borohydride (0.19g, 0.90mmol).After at room temperature stirring is spent the night, dilute this reactant mixture, use the salt water washing, use dried over sodium sulfate, filter and concentrate with dichloromethane.Obtaining required product by the purified by flash chromatography residue, is 4 kinds of mixture of isomers.With regard to 4 kinds of isomers, C ₂₄H ₃₇F ₃N ₅O ₃The LCMS value of calculation: (M+H) 500.3; Measured value 500.3.

Embodiment 18

N-[(1R, 3S)-3-isopropyl-3-(4-[6-methyl-4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

Steps A

1-[6-methyl-4-(trifluoromethyl) pyridine-2-yl] piperazine

With 2-chloro-6-methyl-4-(trifluoromethyl) pyridine (1.0g, 5.11mmol), piperazine (1.32g, 15.3mmol) and triethylamine (0.71mL, solution 5.1mmol) be 1, mixes in the 4-two  alkane (10mL).100 ℃ down stir 5 hours after, this reaction solution of dilute with water and with ethyl acetate extraction 3 times use dried over sodium sulfate, filtration and concentrated in a vacuum.By the thick residue of flash column chromatography purification (10% MeOH/5% Et ₃N/EtOAc) obtain required product (880mg, 70.2%).C ₁₁H ₁₅F ₃N ₃The LCMS value of calculation: (M+H) 246.1; Measured value 246.1.

Step B

[(1R, 3S)-3-isopropyl-3-(4-[6-methyl-4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate

To 1-[6-methyl-4-(trifluoromethyl) pyridine-2-yl] piperazine (280mg, 1.1mmol), (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-isopropyl cyclopentane-carboxylic acid (460mg, 1.7mmol) add benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (0.60g in the solution in dichloromethane (30mL), 1.4mmol) and triethylamine (0.20g, 2.0mmol).After stirring is spent the night, dilute this reactant mixture with dichloromethane, use the salt water washing, use dried over sodium sulfate, filter and concentrate.Obtain required product (200mg, 35.1%) by the thick residue of flash column chromatography purification.C ₂₅H ₃₇F ₃N ₄O ₃The LCMS value of calculation: (M+H) 499.3; Measured value 499.2.

Step C

(1R, 3S)-3-isopropyl-3-(4-[6-methyl-4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) Aminocyclopentane

To be dissolved in 4M 1, HCl (10mL in the 4-two  alkane, 40mmol) [(1R, 3S)-3-isopropyl-3-(4-[6-methyl-4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] (200mg, solution 1.65mmol) at room temperature stirred 1 hour t-butyl carbamate.Dilute this reaction system with dichloromethane, use saturated NaHCO ₃Solution washing is used dried over sodium sulfate, filters and concentrates in a vacuum and obtain required product (0.15g, 94%).C ₂₀H ₃₀F ₃N ₄The LCMS value of calculation of O: (M+H) 399.2; Measured value 399.2.

Step D

N-[(1R, 3S)-3-isopropyl-3-(4-[6-methyl-4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-and 3-isopropyl-3-(4-[6-methyl-4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) Aminocyclopentane (120mg, 0.30mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (120mg, 0.90mmol) and triethylamine (0.12mL, 0.90mmol) add in the solution in dichloromethane (20mL) sodium triacetoxy borohydride (0.19g, 0.90mmol).After at room temperature stirring is spent the night, dilute this reactant mixture, use the salt water washing, use dried over sodium sulfate, filter and concentrate with dichloromethane.Obtain required product by the purified by flash chromatography residue, be the cis/trans mixture of isomers.With regard to 2 kinds of isomers, C ₂₆H ₄₀F ₃N ₄O ₃The LCMS value of calculation: (M+H) 513.3; Measured value 513.2.

Embodiment 19

(4R)-N-[(1R, 3S)-3-isopropyl-3-(4-[3-(trifluoromethyl) phenyl] piperidines-1-base carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine

With (4R)-N-((1R, 3S)-3-isopropyl-3-[4-[3-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] the carbonyl cyclopenta)-(8.0mg 0.016mmol) is dissolved in methanol (0.63mL) to 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine, and N outgases-use ₂Purify, add palladium (3.44mg) (10% dry weight on wet active carbon) subsequently.N outgases reaction flask-uses ₂Purify 3 times, then at room temperature and H ₂(1atm) stirring is spent the night in the environment.This mixture is filtered by the celite pad, with washed with dichloromethane and be concentrated into and obtain required product, be white solid (5.7mg, 71%).C ₂₇H ₄₀F ₃N ₂O ₃The LC-MS value of calculation: (M+H) 497.2; Measured value 497.2.

Embodiment 20

2-[(1R, 3S)-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino]-1-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] preparation of two (trifluoroacetate) (salt) of propan-2-ol

Steps A

(1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-(1-hydroxyl-1-Methylethyl) ring penta-2-alkene-1-methyl formate

In the solution (45mL) of 1.00M hexamethyl two silicon lithium nitrides (Lithiumhexamethyldisilazide) in oxolane that stirs down at-78 ℃, add (1R, 4S)-and the 4-[(tertbutyloxycarbonyl) amino] ring penta-2-alkene-1-methyl formate (5.0g, 21mmol) solution in oxolane (40mL).With the golden mixture temperature of gained-28 ℃ to-23 ℃ (CCl extremely ₄/ dry ice) and stirred 30 minutes.With this reaction solution be cooled to-78 ℃ and add anhydrous propanone (1.8mL, 25mmol).After interpolation, this reactant mixture is remained on CCl ₄To room temperature, spend the night in/the dry ice bath and with its temperature.Use saturated NH ₄Cl makes the dark solution quencher and with ether extraction 3 times.The dry extract that merges filters, and concentrates.Obtain required product (1.4g, 22.6%) by the thick residue of flash column chromatography purification (EtOAc/ hexane).

Step B

(3S)-and the 3-[(tertbutyloxycarbonyl) amino]-1-(1-hydroxyl-1-Methylethyl) cyclopentane-carboxylic acid methyl ester

In the Parr flask with (4S)-4-[(tertbutyloxycarbonyl) amino]-(1.4g 4.7mmol) is dissolved in ethanol (30mL) and use N to 1-(1-hydroxyl-1-Methylethyl) ring penta-2-alkene-1-methyl formate ₂Purify.Add 10% palladium/carbon (0.14g) and the jolting in the nitrogen environment under the 50psi of this mixture is spent the night.This mixture is filtered by celite, with washed with dichloromethane and be concentrated into and obtain required product (1.06g, 86%).

Step C

(3S)-and the 3-[(tertbutyloxycarbonyl) amino]-1-(1-hydroxyl-1-Methylethyl) cyclopentane-carboxylic acid

To (3S)-3-[(tertbutyloxycarbonyl) amino]-1-(1-hydroxyl-1-Methylethyl) cyclopentane-carboxylic acid methyl ester (1.0g, 3.3mmol) (0.22g is 5.3mmol) and with this mixture spend the night (110 ℃) that reflux to add lithium hydroxide monohydrate in the solution in the mixture of oxolane (30mL), methanol (30mL) and water (6mL).The evaporation organic solvent and with ether with water layer washing 1 time.With 6N HCl water layer is acidified to about pH4 then and with dichloromethane extraction 3 times.The dry extract that merges filters and is concentrated into and obtains required product (0.47g, 49%).

Step D

[3-(1-hydroxyl-1-Methylethyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate

To at N ₂(3S)-3-[(tertbutyloxycarbonyl in the environment) amino]-1-(1-hydroxyl-1-Methylethyl) cyclopentane-carboxylic acid (150mg, 0.52mmol) and 1-[4-(trifluoromethyl) pyridine-2-yl] piperazine (130mg, 0.57mmol) add triethylamine (0.16g in the suspension in dichloromethane (3mL), 1.6mol) and benzotriazole-1-base oxygen base three (dimethylamino) phosphorus hexafluorophosphate (0.25g, 0.57mmol).After at room temperature stirring is spent the night, use saturated NaHCO ₃Solution makes the reaction quencher and with dichloromethane extraction 3 times.Dry extract (the MgSO that merges ₄), filter, concentrate and obtain required product (76mg, 29%) by the flash column chromatography purification.

Step e

2-[(1R, 3S)-3-amino-1-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] the propan-2-ol dihydrochloride

Will [(3R)-and 3-(1-hydroxyl-1-Methylethyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] (75mg 0.15mmol) mixes with the solution (2mL) and the oxolane (1mL) of 2.00M hydrogen chloride in ether t-butyl carbamate.After at room temperature stirring 1 hour, this reaction solution is concentrated into obtains required product (70mg, 98.7%).C ₁₇H ₂₈F ₃N ₄O ₂The LCMS value of calculation: (M+H) 473.2; Measured value 473.2.

Step F

2-[(1R, 3S)-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino]-1-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] propan-2-ol two (trifluoroacetate)

To 2-[(1R, 3S)-and 3-amino-1-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] propan-2-ol dihydrochloride (71mg, 0.15mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (69mg, 0.45mmol) and triethylamine (63 μ L, 0.45mmol) add in the solution in dichloromethane (2mL) sodium triacetoxy borohydride (64mg, 0.30mmol).After at room temperature stirring is spent the night, use saturated NaHCO ₃Make the reaction quencher and with dichloromethane extraction 3 times.Dry extract (the MgSO that merges ₄), filter, concentrate, by chromatography purification and change into required product tfa salt (57mg, 57.5%) then.C ₂₅H ₃₇F ₃N ₄O ₄The LCMS value of calculation: (M+H) 515.3; Measured value 515.4.

Embodiment 21

2-[(1S, 3R)-3-[(4R)-3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl] amino-1-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta] preparation of propan-2-ol two (trifluoroacetate)

Use and embodiment 20 described similar operation steps are prepared title compound.C ₂₄H ₃₆F ₃N ₅O ₄The MS value of calculation: (M+H) 516.3; Measured value 516.4.

Embodiment 22

2-[(1S, 3S)-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino]-1-(4-[6-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] preparation of propan-2-ol two (trifluoroacetate)

Use and embodiment 20 described similar operation steps are prepared title compound.C ₂₅H ₃₇F ₃N ₄O ₄The MS value of calculation: (M+H) 515.3; Measured value 515.4.

Embodiment 23

N-[(1S, 3S)-3-ethyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

Steps A

(1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-ethyl ring penta-2-alkene-1-methyl formate

-78 ℃ down and in 10 minutes to the solution (61.5mL of 1.00M hexamethyl two silicon lithium nitrides in oxolane, 61.5mmol) middle (1R that adds, 4S)-and the 4-[(tertbutyloxycarbonyl) amino] ring penta-2-alkene-1-methyl formate (6.71g, 27.8mmol) solution in oxolane (10.0mL).The gained light brown solution was stirred 30 minutes down at-78 ℃, after this add for 1 time iodoethane (2.67mL, 33.4mmol).Then this mixture is remained under-25 ℃ and spend the night.Use saturated NH ₄The Cl aqueous solution makes the reaction quencher.Separate organic layer and further water layer is extracted 3 times with ether.Use the organic layer of above many merging then, use Na ₂SO ₄, drying is filtered, and concentrate and obtain required product by the flash column chromatography purification, be 7: 1 cis/trans mixture (4.83g, 65%).C ₁₄H ₂₃NO ₄The MS value of calculation: (M+H) 170.2; Measured value 170.1 (M+H-Boc).

Step B

(1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-ethyl ring penta-2-alkene-1-formic acid

To (1R, 4S)-and the 4-[(tertbutyloxycarbonyl) amino]-1-ethyl ring penta-2-alkene-1-methyl formate (4.80g, 17.8mmol)) (1.2g refluxes 28.6mmol) and with this mixture and to spend the night to add lithium hydroxide monohydrate in the solution in the mixture of oxolane (100mL), methanol (100mL) and water (20mL).The evaporation organic solvent.With 6N HCl water layer is acidified to about pH4 then and with dichloromethane extraction 3 times.The dry extract that merges filters and is concentrated into and obtains cis/trans isomer mixture (2.93g, cis/trans=7: 1), is faint yellow solid.This solid is dissolved in EtOAc (4.0mL), obtains settled solution by heating and with hexane dilution (100mL).In 1 hour, this solution slowly cooled to room temperature and maintain then under-25 ℃ and spend the night.Make cis-isomer crystallizes and be dried to obtain required product (1.40g, 31%), be white solid.C ₁₃H ₂₁NO ₄The MS value of calculation: (M+H) 256.2.2; Measured value 156.1 (M+H-Boc).

Step C

(1S, 3R)-the 3-[(tertbutyloxycarbonyl) amino]-1-ethyl cyclopentane formic acid

To (1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-(1.38g 5.41mmol) adds 10% palladium/carbon (200mg) to 1-ethyl ring penta-2-alkene-1-formic acid in the solution in ethanol (40mL).With the jolting 18 hours and filter in the hydrogen environment under the 50psi of this mixture by celite.Evaporated filtrate and obtain required product (1.5g) in a vacuum.C ₁₃H ₂₃NO ₄The MS value of calculation: (M+H) 258.2; Measured value 158.1 (M+H-Boc).

Step D

[(1S, 3R)-3-ethyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate

To at N ₂(1R in the environment, 3S)-and the 3-[(tertbutyloxycarbonyl) amino]-1-ethyl cyclopentane formic acid (0.30g, 1.2mmol) and 1-[4-(trifluoromethyl) pyridine-2-yl] piperazine dihydrochloride (0.39g, 1.3mmol) add triethylamine (0.65mL in the solution in DMF (10mL), 4.7mmol) and O-(benzotriazole-1-yl)-N, N, N ', N '-tetramethylurea hexafluorophosphate (0.663g, 1.75mmol).After at room temperature stirring is spent the night, use saturated NaHCO ₃Make the reaction quencher and with dichloromethane extraction 3 times.Dry extract (the MgSO that merges ₄), filter, concentrate and obtain required product (400mg, 72.9%) by the flash column chromatography purification.C ₂₃H ₃₄F ₃N ₄O ₃The MS value of calculation: (M+H) 471.3; Measured value 371.2 (M+H-Boc).

Step e

(1S, 3R)-3-ethyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) the Aminocyclopentane dihydrochloride

With [(1S, 3R)-and 3-ethyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate (0.39g, 0.83mmol) be dissolved in 4M hydrogen chloride 1, the solution (3.1mL) in the 4-two  alkane and this solution at room temperature stirred spend the night.Evaporation reaction solution and obtain required product is yellow powder (0.36g, 96%).

Step F

N-[(1S, 3S)-3-ethyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

To (1S, 3S)-and 3-ethyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) Aminocyclopentane dihydrochloride (100mg, 0.20mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (77mg, 0.50mmol) and triethylamine (0.110mL, 0.79mmol) add in the solution in dichloromethane (3mL) sodium triacetoxy borohydride (96mg, 0.45mmol).After at room temperature stirring is spent the night, use saturated NaHCO ₃Make the reaction quencher and with dichloromethane extraction 3 times.Dry organic layer (the MgSO that merges ₄), filter, concentrate, by column chromatography purification (NH ₄OH/MeOH/EtOAc) obtain required product (111mg, 69.1%).C ₂₄H ₃₅F ₃N ₄O ₃The MS value of calculation: (M+H) 485.3; Measured value 485.2.

Embodiment 24

(4R)-N-[(1R, 3S)-3-ethyl-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

Use and embodiment 23 described similar operation steps are prepared title compound.C ₂₃H ₃₄F ₃N ₅O ₃The MS value of calculation: (M+H) 486.3; Measured value 486.2.

Embodiment 25

N-[(1S, 3S)-3-ethyl-3-(4-[6-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

Use and embodiment 23 described similar operation steps are prepared title compound.C ₂₄H ₃₅F ₃N ₄O ₃The MS value of calculation: (M+H) 485.3; Measured value 485.3.

Embodiment 26

(4R)-N-[(1R, 3S)-3-methyl-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

Use and embodiment 23 described similar operation steps are prepared title compound.C ₂₂H ₃₂F ₃N ₅O ₃The MS value of calculation: (M+H) 472.3.3; Measured value 472.3.

Embodiment 27

(4R)-3-methoxyl group-N-[(1R, 3S)-3-(2-methoxy ethyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] preparation of tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

Steps A

1-iodo-2-Ethyl Methyl Ether

To 1-bromo-2-Ethyl Methyl Ether (2.0g, 14mmol) add in the solution in acetone (40mL) sodium iodide (11g, 72mmol) and with gained solution at N ₂Backflow (70 ℃) is 3 hours in the environment.Cool off this mixture and filtration.When in cold closet, further cooling off, there is extra solid to separate out and they are filtered out, after this is concentrated into and obtains orange residue.Residue is dissolved in ether and uses Na ₂S ₂O ₃Washing produces near colourless solution.Dry this solution (MgSO ₄), filter and be concentrated into and obtain yellow oil (1.8g, 64%). ¹H?NMR(CDCl ₃)δ3.70-3.60(2H，t，J＝5Hz)，3.40(3H，s)，3.30-3.20(2H，t，J＝5Hz)。

Step B

(1S, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-(2-methoxy ethyl) ring penta-2-alkene-1-methyl formate

To descending and N at-78 ℃ ₂The solution (9.1mL of 1.00M hexamethyl two silicon lithium nitrides in oxolane in the environment, 9.1mmmol) middle (1R that adds, 4S)-and the 4-[(tertbutyloxycarbonyl) amino] ring penta-2-alkene-1-methyl formate (1.0g, 4.1mmol) solution in oxolane (2.0mL).The gained light brown solution was stirred 30 minutes down at-78 ℃, after this add 1-iodo-2-Ethyl Methyl Ether (0.93g, 5.0mmol) solution in oxolane (2.0mL).The gained mixture was stirred 1 hour down at-78 ℃, remain on then in the cold closet of-20 ℃ of readings and spend the night.Make the reaction quencher with saturated ammonium chloride.Separate each layer and water layer extract is extracted 3 times with ether.The organic layer that merges with the salt water washing then, dry (magnesium sulfate) filters and obtains required product (0.28g, 23%) by purified by flash chromatography (EtOAc/ hexane).C ₁₅H ₂₆NO ₅The LCMS value of calculation: (M+H) 300.2; Measured value 300.2.

Step C

(1S, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-(2-methoxy ethyl) ring penta-2-alkene-1-formic acid

To (the 1S that stirs, 4S)-and the 4-[(tertbutyloxycarbonyl) amino]-1-(2-methoxy ethyl) ring penta-2-alkene-1-methyl formate (0.78g, 2.6mmol) (0.55g stirs down at 80 ℃ 13mmol) and with the gained orange mixture and to spend the night to add lithium hydroxide monohydrate in the solution in oxolane (1.5mL), methanol (15mL) and water (3.0mL).Evaporating solvent and with 6NHCl this mixture is acidified to pH and is about 4.With dichloromethane water layer is extracted 3 times then.Dry organic layer (the MgSO that merges ₄), filter and be concentrated in a vacuum and obtain required product, be grease (0.47g, 63.2%).C ₁₄H ₂₄NO ₅The LCMS value of calculation: (M+H) 286.2; Measured value 286.2.

Step D

(1S, 3R)-the 3-[(tertbutyloxycarbonyl) amino]-1-(2-methoxy ethyl) cyclopentane-carboxylic acid

To (1S, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-(1.56g 5.47mmol) adds 10% palladium/carbon (150mg) to 1-(2-methoxy ethyl) ring penta-2-alkene-1-formic acid in the solution in methanol (30mL).The jolting in the hydrogen environment under the 50psi of this mixture is spent the night and filter by celite.Evaporated filtrate and obtain required product (1.57g, 99.9%) in a vacuum.C ₁₄H ₂₆NO ₅MS value of calculation (M+H) 288.2; Measured value 188.2 (M+H-Boc).

Step e

[(1R, 3S)-3-(2-methoxy ethyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate

With (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-(2-methoxy ethyl) cyclopentane-carboxylic acid (276.6mg, 0.96mmol), 1-[4-(trifluoromethyl) pyridine-2-yl] piperazine dihydrochloride (322.0mg, 1.06mmol), triethylamine (0.54mL, 3.85mmol) and O-(benzotriazole-1-yl)-N, N, N ', (547.6mg, 1.44mmol) (HBTU) mixes in dry DMF (6.6mL) and with the gained brown solution at room temperature and N N '-tetramethylurea hexafluorophosphate ₂Stirred 3 days in the environment.Use CH ₂Cl ₂Diluted reaction mixture and use saturated Na ₂CO ₃Washing.Use CH ₂Cl ₂Water layer is extracted 4 times.Dry organic layer (the MgSO that merges ₄), filter, concentrate and obtain required product (252mg, 52%) by purified by flash chromatography (EtOAc/ hexane).C ₂₄H ₃₆F ₃N ₄O ₄MS value of calculation (M+H) 501.3; Measured value 401.3 (M+H-Boc).

Step F

(1R, 3S)-3-(2-methoxy ethyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) the Aminocyclopentane dihydrochloride

With [(1R, 3S)-3-(2-methoxy ethyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate (252mg, 0.503mmol) be dissolved in 2M hydrogen chloride in ether solution (8mL) and at room temperature stirred 2 hours.This reaction solution is concentrated into obtains required product, be yellow powder (0.36g, 96%).C ₁₉H ₂₇F ₃N ₄O ₂MS value of calculation (M+H) 401.3; Measured value 401.3.

Step G

(4R)-3-methoxyl group-N-[(1R, 3S)-3-(2-methoxy ethyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

To (1R, 3S)-and 3-(2-methoxy ethyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) Aminocyclopentane dihydrochloride (140.0mg, 0.30mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (115mg, 0.887mmol) and triethylamine (0.165mL, 1.18mmol) add in the solution in dry methylene chloride (12mL) sodium triacetoxy borohydride (188.1mg, 0.887mmol).At room temperature and N ₂After stirring is spent the night in the environment, use saturated NaHCO ₃Make the reaction quencher and with dichloromethane extraction 3 times.Dry organic layer (the MgSO that merges ₄), filter, concentrate, by flash column chromatography purification (MeOH/EtOAc) and change into tfa salt and obtain required product (72.4mg, 34%).C ₂₅H ₃₇F ₃N ₄O ₄The MS value of calculation: (M+H) 515.3; Measured value 515.4.

Embodiment 28

3-methoxyl group-N-[(1S, 3S)-3-(2-methoxy ethyl)-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta] preparation of tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

Use and embodiment 27 described similar operation steps are prepared title compound.C ₂₄H ₃₆F ₃N ₅O ₄The MS value of calculation: (M+H) 516.3; Measured value 516.3.

Embodiment 29

(4R)-N-[(1R, 3S)-3-(ethoxyl methyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

Steps A

(1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-(ethoxyl methyl) ring penta-2-alkene-1-methyl formate

In the solution (36.7mL) of 1.0M hexamethyl two silicon lithium nitrides, adding (1R, 4S)-4-[(tertbutyloxycarbonyl) amino under-78 ℃ at oxolane] ring penta-2-alkene-1-methyl formate (4.00g, 1.66mmol) solution in oxolane (6.0mL).The gained light brown solution was stirred 30 minutes down at-78 ℃, after this add for 1 time (chlorine methoxyl group) ethane (1.88g, 19.9mol).This mixture was stirred 1 hour down and keeps spending the night then in the cold closet of reading under-25 ℃ at-78 ℃.Use saturated NH then ₄Cl (50mL) makes the reaction quencher.Separate organic layer and use CH ₂Cl ₂Water layer is extracted 3 times.With the organic layer that above washing merges, use Na ₂SO ₄Drying is filtered, and concentrate and obtain required product (3.29g, 66%) by purified by flash chromatography (EtOAC of 0-15% in hexane), be cis/trans (3: 2) mixture based on the reversed-phase HPLC analysis.C ₁₅H ₂₆NO ₅The MS value of calculation: (M+H) 300.2; Measured value 200.2 (M+H-Boc).

Step B

(1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-(ethoxyl methyl) ring penta-2-alkene-1-formic acid

To (1R, 4S)-and the 4-[(tertbutyloxycarbonyl) amino]-1-(ethoxyl methyl) ring penta-2-alkene-1-methyl formate (3.25g, 10.8mmol) add in the solution in oxolane (58.7mL), methanol (58.7mL) and water (12.6mL) lithium hydroxide monohydrate (0.731g, 17.42mmol).Should the pink mixture heated spend the night to refluxing.Remove organic solvent in a vacuum and with ether with water layer washing 1 time and slowly be acidified to pH with dense HCl then and reach 4.With dichloromethane the gained suspension is extracted 3 times.Use MgSO ₄The dry organic layer that merges filters and is concentrated into and obtains required product, is cis/trans isomer mixture (2.75g, 89%).C ₁₄H ₂₄NO ₅The MS value of calculation: (M+H) 286.2; Measured value 186.2 (M+H-Boc).

Step C

(1S, 3R)-the 3-[(tertbutyloxycarbonyl) amino]-1-(ethoxyl methyl) cyclopentane-carboxylic acid

To (1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-(2.70g 9.46mmol) adds 10% palladium/carbon (350mg) to 1-(ethoxyl methyl) ring penta-2-alkene-1-formic acid in the solution in ethanol (69.5mL).With the jolting 18 hours in the hydrogen environment under the 50psi of this mixture, filter and use washed with dichloromethane by celite.Concentrated filtrate and obtain required product (2.87g).C ₁₄H ₂₆NO ₅MS value of calculation (M+H) 288.2; Measured value 188.2 (M+H-Boc).

Step D

[(3S)-and 3-(ethoxyl methyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate

With (1S)-3-[(tertbutyloxycarbonyl) amino]-1-(ethoxyl methyl) cyclopentane-carboxylic acid (429.4mg, 1.494mmol), 1-[4-(trifluoromethyl) pyridine-2-yl] piperazine dihydrochloride (500.0mg, 1.644mmol), triethylamine (0.833mL, 5.98mmol) and O-(benzotriazole-1-yl)-N, N, N ', (850.3mg, 2.242mol) (HBTU) mixes in dry DMF (10.2mL) N '-tetramethylurea hexafluorophosphate.With the gained brown solution at room temperature and N ₂Stir in the environment and spend the night.Use CH ₂Cl ₂Diluted reaction mixture and use saturated Na ₂CO ₃Washing.Use CH ₂Cl ₂Water layer is extracted 4 times.Dry organic layer (the MgSO that merges ₄), concentrate and obtain required product (304.4mg, 40%) by purified by flash chromatography.C ₂₄H ₃₆F ₃N ₄O ₄The MS value of calculation: (M+H) 501.3; Measured value 501.3.

Step e

(1R, 3S)-3-(ethoxyl methyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) the Aminocyclopentane dihydrochloride

Will [(3S)-and 3-(ethoxyl methyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] (295mg 0.589mol) is dissolved in the solution (10mL) of 2M hydrogen chloride in ether to t-butyl carbamate.At room temperature stir spend the night after, concentrate this reactant mixture in a vacuum and obtain required product (330mg), be faint yellow solid.C ₁₉H ₂₈F ₃N ₄O ₂The MS value of calculation: (M+H) 401.3; Measured value 401.3.

Step F

(4R)-N-[(1R, 3S)-3-(ethoxyl methyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

To (1R, 3S)-and 3-(ethoxyl methyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) Aminocyclopentane dihydrochloride (100.0mg, 0.211mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (82.5mg, 0.634mmol) and triethylamine (0.118mL, 0.845mmol) add in the solution in dry methylene chloride (8.6mL) sodium triacetoxy borohydride (134.3mg, 0.634mmol).At room temperature and N ₂After stirring is spent the night in the environment, use saturated NaHCO ₃Aqueous solution makes the reaction quencher and dilutes with dichloromethane.Separate organic layer and water layer is extracted 3 times with dichloromethane.Merge organic layer, use MgSO ₄Drying is filtered, by silica gel combination-rapid system (12 restrain posts for gradient, the MeOH of 0-40% in EtOAc) purification and change into tfa salt and obtain required product (115.4mg, 74%).C ₂₅H ₃₇F ₃N ₄O ₄The MS value of calculation: (M+H) 515.3; Measured value 515.4.

Embodiment 30

(4R)-N-[(1R, 3S)-3-(ethoxyl methyl)-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta]-preparation of 3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

Use and embodiment 29 described similar operation steps are prepared title compound.C ₂₄H ₃₆F ₃N ₅O ₄The MS value of calculation: (M+H) 516.3; Measured value 516.4.

Embodiment 31

(4R)-3-methoxyl group-N-[(1R, 3S)-3-(methoxy)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] preparation of tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

Use and embodiment 29 described similar operation steps are prepared title compound.C ₂₄H ₃₅F ₃N ₄O ₄The MS value of calculation: (M+H) 501.3; Measured value 501.3.

Embodiment 32

(4R)-3-methoxyl group-N-[(1R, 3S)-3-(methoxy)-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta] preparation of tetrahydrochysene-2H-pyrans-4-amine two (trifluoroacetate)

Use and embodiment 29 described similar operation steps are prepared title compound.C ₂₃H ₃₄F ₃N ₅O ₄The MS value of calculation: (M+H) 502.3; Measured value 502.3.

Embodiment 33

(4R)-3-methoxyl group-N-[(1R, 3S)-3-[(3R)-oxolane-3-yl]-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] preparation of tetrahydrochysene-2H-pyrans-4-amine

Steps A

(3R)-3-iodine oxolane

To (S)-(+)-3-hydroxyl tetrahydrofuran (0.50g, 5.7mmol) add successively in the solution in dichloromethane (50mL) triphenylphosphine (3.0g, 11mmol), the 1H-imidazoles (0.75g, 11mmol) and iodine (2.9g, 11mmol).At N ₂After backflow is spent the night in the environment, use 0.2MNa ₂S ₂O ₃(60mL) make the reaction quencher.Separate organic layer and water layer is extracted 3 times with dichloromethane.Dry organic layer (the MgSO that merges ₄), filter and be concentrated into the yellow solid that obtains wetting.In this solid, add pentane (100mL) and stirred 2 hours.Filter out solid and concentrated filtrate and obtain required product (970mg, 79.4%), be yellow oil. ¹H?NMR(CDCl ₃)δ4.30-3.85(5H，m)，2.50-2.20(2H，m)。

Step B

(1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-[(3R)-and oxolane-3-yl] ring penta-2-alkene-1-methyl formate

To descending and N at-78 ℃ ₂The solution (34.8mL of 1.00M hexamethyl two silicon lithium nitrides in oxolane in the environment, 34.8mmol) middle (1R that adds, 4S)-and the 4-[(tertbutyloxycarbonyl) amino] ring penta-2-alkene-1-methyl formate (4.0g, 16mmol) solution in oxolane (20mL).The gained brown solution was stirred 30 minutes down at-78 ℃, after this add (3R)-3-iodine oxolane (3.75g, 17.4mmol) solution in THF (3mL).This mixture was stirred 10 minutes down at-78 ℃, keep then spending the night in the cold closet of reading under-25 ℃.Make the reaction quencher with saturated ammonium chloride, use ether extraction 3 times.Dry organic extract (the MgSO that merges ₄), filter, concentrate and obtain required product (1.6g, 31%) by flash column chromatography purification (EtOAc/ hexane).C ₁₆H ₂₅NO ₅The MS value of calculation: (M+H) 312.2; Measured value 312.2.

Step C

(1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-[(3R)-and oxolane-3-yl] ring penta-2-alkene-1-formic acid

To (1R, 4S)-and the 4-[(tertbutyloxycarbonyl) amino]-1-[(3R)-and oxolane-3-yl] ring penta-2-alkene-1-methyl formate (1.60g, 5.14mmol) add in the solution in oxolane (27.8mL), methanol (27.8mL) and water (6.0mL) lithium hydroxide monohydrate (0.346g, 8.25mmol).Should the pink mixture heated to refluxing 18 hours.Remove organic solvent in a vacuum and with ether with water layer washing 1 time and slowly be acidified to pH with 6M HCl then and reach 3-4.Use CH ₂Cl ₂The gained suspension is extracted 3 times.Use MgSO ₄The dry organic layer that merges filters and is concentrated into and obtains product, is 2 kinds of cis/trans isomer mixtures (1.59g), is faint yellow solid.C ₁₅H ₂₄NO ₅The MS value of calculation: (M+H) 298.2; Measured value 198.2 (M+H-Boc).

Step D

(1S, 3R)-the 3-[(tertbutyloxycarbonyl) amino]-1-[(3R)-and oxolane-3-yl] cyclopentane-carboxylic acid

Will (1R, 4S)-the 4-[(tertbutyloxycarbonyl) amino]-1-[(3R)-and oxolane-3-yl] alkene-(0.79g 2.6mol) is dissolved in ethanol (20.0mL) to 2-formic acid to ring penta-2-, and N outgases-use ₂Purify, add subsequently platinum dioxide (0.150g, 0.528mol).This reactant mixture put into the Parr instrument and at 55psi H ₂Following hydrogenation 18 hours.This mixture is filtered by the celite pad,, be concentrated into and obtain required product (730mg, 91.8%) with the MeOH washing.C ₁₅H ₂₆NO ₅MS value of calculation (M+H) 300.2; Measured value 200.2 (M+H-Boc).

Step e

[(1R, 3S)-3-[(3R)-oxolane-3-yl]-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] t-butyl carbamate

With (1S, 3R)-and the 3-[(tertbutyloxycarbonyl) amino]-1-[(3R)-and oxolane-3-yl] cyclopentane-carboxylic acid (350.0mg, 1.169mmol), 1-[4-(trifluoromethyl) pyridine-2-yl] piperazine dihydrochloride (391.1mg, 1.286mmol), triethylamine (0.652mL, 4.68mmol) and O-(benzotriazole-1-yl)-N, N, N ', (665.1mg, 1.754mol) (HBTU) mixes in dry DMF (8.0mL) N '-tetramethylurea hexafluorophosphate.After at room temperature stirring is spent the night, use CH ₂Cl ₂Diluted reaction mixture and use saturated Na ₂CO ₃Washing.Use CH ₂Cl ₂Water layer is extracted 4 times.Dry organic layer (the MgSO that merges ₄), concentrate and obtain required product (270mg, 45%) by purified by flash chromatography (40 restrain posts for combination-rapid system, the 0-50% EtOAcs in hexane, gradient elution).C ₂₅H ₃₆F ₃N ₄O ₄MS value of calculation (M+H) 513.3; Measured value 513.3.

Step F

(1R, 3S)-3-[(3R)-oxolane-3-yl]-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) the Aminocyclopentane dihydrochloride

Will [(1R, 3S)-3-[(3R)-oxolane-3-yl]-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] (260mg 0.51mmol) is dissolved in the solution (8mL) of 2M hydrogen chloride in ether to t-butyl carbamate.At room temperature stir spend the night after, concentrate this reactant mixture in a vacuum and obtain required product (290mg), be faint yellow solid.C ₂₀H ₂₇F ₃N ₄O ₂The MS value of calculation: (M+H) 413.2; Measured value 413.0.

Step G

(4R)-3-methoxyl group-N-[(1R, 3S)-3-[(3R)-oxolane-3-yl]-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] tetrahydrochysene-2H-pyrans-4-amine

To (1R, 3S)-3-[(3R)-oxolane-3-yl]-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) Aminocyclopentane dihydrochloride (73.8mg, 0.152mmol), 3-methoxyl group tetrahydrochysene-4H-pyrans-4-ketone (59.4mg, 0.456mmol) and triethylamine (0.0848mL, 0.608mmol) add in the solution in dry methylene chloride (4.1mL) sodium triacetoxy borohydride (96.7mg, 0.456mmol).At room temperature and N ₂After stirring in the environment, use NaHCO ₃Aqueous solution makes the reaction quencher and uses CH ₂Cl ₂Dilution.Separate organic layer and use CH ₂Cl ₂Water layer is extracted 3 times.Merge organic layer, use MgSO ₄Drying is filtered, and concentrates and obtains required product (20mg, 41%) by silica gel chromatography purification (12 restrain posts for combination-rapid system, the 0-40% MeOH in EtOAc, gradient).C ₂₆H ₃₇F ₃N ₄O ₄The MS value of calculation: (M+H) 527.3; Measured value 527.3.

Embodiment 34

(4R)-3-methoxyl group-N-[(1R, 3S)-3-[(3R)-oxolane-3-yl]-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta] preparation of tetrahydrochysene-2H-pyrans-4-amine

Use and embodiment 33 described similar operation steps are prepared title compound.C ₂₅H ₃₆F ₃N ₅O ₄The MS value of calculation: (M+H) 528.3; Measured value 528.3.

Embodiment A

The CCR2 in vitro tests

Can use suitable screening (for example high throughput test) to measure the ability of noval chemical compound antagonism chemokine receptors of the present invention (for example CCR2) function.For example, extracellular acidification test, the test of calcium current flux, part in conjunction with test or chemotaxis test in test agents (for example, referring to Hesselgesser etc., J Biol.Chem.273 (25): 15687-15692 (1998); WO00/05265 and WO 98/02151).

In suitable test, use can separation or reorganization the CCR2 albumen in source, it has the proteic characteristic of at least a mammal CCR2, activity or functional character.Specificity can be associativity (for example combining with part or inhibitor), signaling activity (for example mammal G protein activation, kytoplasm free calcium [Ca ⁺⁺] the inducing of quick and instantaneous increase, cell effect function (for example leukocyte stimulation chemotaxis or mediator of inflammation discharge) etc. of i concentration.

In in conjunction with test examples, the compositions that will contain CCR2 albumen or its variant maintains and is suitable under the bonded condition.Make CCR2 receptor engaged test chemical compound and detection or measure combination.

In example, use the carrier of using nucleotide sequence or express the stable or of short duration cells transfected of cartridge clip with coding CCR2 receptor based on the test of cell.Maintain cell under the condition that is suitable for expression of receptor and make it under being suitable for, contact activating agent in conjunction with the condition that takes place.Can use standard technique to detect combination.For example, measure the combination degree of comparing with suitable matched group.In addition, can use the cell part that contains described receptor, replace intact cell such as membrane portions.

Form and directly or indirectly to detect in conjunction with the complex in detecting or testing.For example, can use suitable labelling (for example fluorescent labeling, isotopic labeling, enzyme labelling etc.) labelling activating agent and can be by detecting this marker determination combination.Can use unlabelled activating agent or part to estimate specificity and/or competitive combination by competitive or replacement research as competitor.

The CCR2 antagonist activities of The compounds of this invention can be reported to and reach specificity in the receptor binding assays in conjunction with the required inhibitor concentration (IC of 50% inhibition ₅₀Be worth), described test is used ¹²⁵The MCP-1 of I-labelling is as part with by the peripheral blood lymphocytes (PBMCs) of normal person's whole blood by the density gradient centrifugation preparation.Preferably specificity is deducted non-specific binding in conjunction with being defined as overall combination (for example total cpm on the filter).Non-specific binding is defined as in the presence of the excessive unlabelled competitor (for example MCP-1) the still amount of detected cpm.

Embodiment B

In conjunction with test

In in conjunction with test, human PBMC s is used to test The compounds of this invention.For example, with 200,000-500,000 cell and 0.1-0.2nM ¹²⁵The MCP-1 of I-labelling, with or not with the testing compound incubation of unlabelled competitor (10nM MCP-1) or various concentration.By suitable method preparation ¹²⁵The MCP-1 of I-labelling or they are available from goods providers (Perkin Elmer, Boston MA).Association reaction at room temperature and in the 50-250 μ L binding buffer liquid of being made up of 1M HEPES pH7.2 and 0.1% BSA (bovine serum albumin) was carried out 30 minutes.Filter the collection film fast by the glass fiber filter (Perkin Elmer) in pre-immersion 0.3% polymine or phosphate-buffered saline (PBS) and stop association reaction.With the about 600 μ L binding buffer liquid flushing filter that contains 0.5M NaCl or PBS, dry then and by measuring bonded radioactive amount with gamma counter (Perkin Elmer) counting.

In conjunction with testing program, chemical compound of the present invention has the IC that is lower than about 3000nM according to this ₅₀Value.

Embodiment C

The chemotaxis test

In the Boyden Chamber (Neuro Probe) that modifies, use peripheral blood lymphocytes in the leucocyte chemotaxis test, to measure the ability of The compounds of this invention antagonism CCR2 function.Will 500,000 cells in not containing the DMEM culture medium of serum (In Vitrogen) with or not with described inhibitor incubation and the temperature to 37 ℃.Also pre-warm chemotactic chamber (NeuroProbe).The 10nM MCP-1 that 400 μ L are warm be heated in the floor chamber in porose, and negative control adds DMEM.8 micron membranes filter membranes (Neuro Probe) are placed on the top and seal with chamber cap.Then cell is joined in the hole in the chamber cap, these holes are connected with hole, chamber under the filter membrane.With whole chamber at 37 ℃, 5% CO ₂Middle incubation 30 minutes.Aspirate out cell then, open chamber cap and slowly remove filter membrane.With PBS filter membrane top washing 3 times and maintenance are not contacted the bottom.Filter membrane is air-dry and dye with Wright Geimsa stain (Sigma).Checking filter membrane by microscope inspection counts.Negative control hole is deducted from all values as background and with it.Compare to determine the antagonist effect by cell quantity that will migrate to floor chamber in the hole of containing antagonist and the cell quantity that migrates to floor chamber in the MCP-1 control wells.

According to this chemotaxis test, chemical compound of the present invention has the IC that is lower than about 3000nM ₅₀Value.

Embodiment D

CCR5 expresses

(Biological Specialty, Colmar is PA) available from the donor that does not contain normally medicine and by density gradient centrifugation separating periphery blood monocytic cell (PBMCs) for leukophoresis.By the further separating monocytic cell of centrifugal elutriation.After washing, use replenished 10% FBS (Hyclone, Logan, UT) and 10-20ng/mL recombined human IL-10 (R﹠amp; Dsystems, Minneapolis, (Invitrogen, Carlsbad is CA) with 10 for RPMI MN) ⁶Suspend again mononuclear cell and down and contain 5% CO of individual cell/ml at 37 ℃ ₂Culture medium in hatched 24-48 hour.Anti-Human CCR5 antibody (PharMingen by puting together then with PE-, San Diego, CA) dyeing verifies that the CCR5 on the mononuclear cell that IL-10-handles expresses, and uses FACSCalibur (BD Biosciences subsequently, Bedford MA) carries out facs analysis.

Embodiment E

CCR5 is in conjunction with test

At 96 hole MultiScreen ^TMFilter plate (Millipore Systems, Billerica, MA) in, at room temperature and contain 20mM HEPES (Invitrogen, Carlsbad, CA) and 0.3%BSA (Sigma, St Louis, MO) 150 μ L RPMI (Invitrogen, Carlsbad, CA) in 3 * 10 ⁵Mononuclear cell and 0.2nM that individual IL-10-handles ¹²⁵I-MIP-1 β (PerkinElmer, Boston, MA) and the The compounds of this invention of a series of concentration incubation 1 hour together.By with cell and 0.3 μ M MIP-1 β (R﹠amp; D Systems, Minneapolis, MN) incubation is measured non-specific binding together.By (Millipore Systems, Billerica MA) stop association reaction on the filter membrane in the flat board at vacuum manifold with cell harvesting.Then on vacuum manifold with replenished 20mM HEPES (Invitrogen, Carlsbad, CA), 0.3% BSA (Sigma, St Louis, MO) and RPMI (Invitrogen, the Carlsbad of 0.4M NaCl, CA), air-dry and from flat board, peel off with filter membrane washing 5 times.(Millipore Systems, Billerica MA) stamp out filter dish (dish) excessively suitable with sample well in the filter plate to use the micropore stamping system.Cross upward bonded radioactive amount of filter dish (dish) by measure each with the gamma counter counting.Specificity is deducted non-specific binding in conjunction with being defined as overall combination.(GraphPad Software, San Diego CA) estimate binding data to use Prism.Find that according to this test chemical compound of the present invention has about 1 μ M or the binding affinity below the 1 μ M.

Embodiment F

HIV-1 enters test

By for example Connor etc. at Virology, 206 (1995), described in the 935-944, with the plasmid (modifying) of the NL4-3 bacterial strain of coding HIV-1 and the plasmid co-transfection of one of several HIV-1 env genes of coding, produce replication defect type HIV-1 report virion by making env gene sudden change and importing luciferase report plasmid.By behind two kinds of plasmids of calcium phosphate precipitation transfection, in the time of the 3rd day, collect viral supernatant and measurement function sexually transmitted disease (STD) poison titre.Then these stock solutions are used to infect stably express CD4 and chemokine receptor CCR 5 with or not with the U87 cell of testing compound precincubation.To infect and under 37 ℃, carry out 2 hours, washed cell and replace with the fresh culture that contains chemical compound.Cell was hatched 3 days dissolving and mensuration uciferase activity.The result is reported to the required compound concentration of 50% uciferase activity in the inhibition control cultures.

Embodiment G

HIV-1 replicated test in the MT-4 cell

Carry out HIV-1 NL4.3 (or III as mentioned above _B) duplicate test (Bridger etc., the J.Med.Chem.42:3971-3981 (1999) of inhibition; De Clercq etc., Proc.Natl.Acad.Sci.89:5286-5290 (1992); De Clercq etc., Antimicrob.AgentsChemother.38:668-674 (1994); Bridger etc., J.Med.Chem.38:366-378 (1995)).For the purpose of summarizing, parallel anti-HIV activity and the cytotoxic assay of carrying out, and based on the MT-4 cell survival rate that has infected HIV in the presence of the variable concentrations test compounds is being arranged.Make the MT-4 cell proliferation after 5 days, by the calorimetric 3-based on tetrazole in 96-hole microarray (microtrays) (4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazole bromide (MTT) operating procedure is carried out quantitatively the quantity of survivaling cell.The result is quantized and obtain EC ₅₀Value, their representatives prevent the required concentration of cell generation pathological changes caused by virus (cytopathicity) of 50% viral infection.

Except that those technical schemes as herein described, apparent to those skilled in the art to various modification of the present invention according to foregoing description.This class modification also belongs to the scope of attached batch of claim.With each list of references of quoting among the application, comprise that the full content of patent, patent application and open source literature is incorporated herein by reference.

Claims

1. the chemical compound of general formula I:

Or its pharmaceutically acceptable salt or prodrug, wherein:

Dotted line is represented the key chosen wantonly;

W is:

V is N, NO or CR ⁵

X is N, NO or CR ²

Y is N, NO or CR ³

R ¹Be C _1-6Alkyl, C _1-6Haloalkyl, C _1-6Hydroxy alkyl ,-(C _0-6Alkyl)-O-(C _1-6Alkyl) ,-(C _0-6Alkyl)-S-(C _1-6Alkyl) ,-(C _0-6Alkyl)-(C _3-7Cycloalkyl)-(C _0-6Alkyl), OH, OR ¹⁰, SR ¹⁰, COR ¹¹, CO ₂R ¹⁰, CONR ¹⁰R ¹², carbocylic radical, heterocyclic radical, CN, NR ¹⁰R ¹², NR ¹⁰SO ₂R ¹⁰, NR ¹⁰COR ¹⁰, NR ¹⁰CO ₂R ¹⁰, NR ¹⁰CONR ¹², CR ¹⁰R ¹¹CO ₂R ¹⁰Or CR ¹⁰R ¹¹OCO ¹⁰

R ⁸Be C _1-3Alkoxyl, C _1-3Halogenated alkoxy, C _3-6Cycloalkyloxy or OH;

R ^{8 '}Be H;

R ¹⁰Be H, C _1-6Alkyl, benzyl, phenyl or C _3-6Cycloalkyl, wherein said C _1-6Alkyl, benzyl, phenyl or C _3-6Cycloalkyl randomly is selected from halogen, OH, C by 1-3 _1-3Alkyl, C _1-3Haloalkyl, C _1-3Alkoxyl, C _1-3Halogenated alkoxy, CO ₂H and CO ₂-(C _1-6Alkyl) substituent group replaces;

R ¹²Be H, C _1-6Alkyl, benzyl, phenyl or C _3-6Cycloalkyl, wherein said C _1-6Alkyl, benzyl, phenyl or C _3-6Cycloalkyl randomly is selected from halogen, OH, C by 1-3 _1-3Alkyl, C _1-3Haloalkyl, C _1-3Alkoxyl, C _1-3Halogenated alkoxy, CO ₂H and CO ₂-(C _1-6Alkyl) substituent group replaces; And

P is 0 or 1.

2. the described chemical compound of claim 1, wherein W is

3. the described chemical compound of claim 1, wherein W is

4. the described chemical compound of claim 1, wherein V is CR ⁵

5. the described chemical compound of claim 1, wherein X is CR ²

6. the described chemical compound of claim 1, wherein Y is CR ³

7. the described chemical compound of claim 1, wherein Z is CR ⁴

8. the described chemical compound of claim 1, wherein X is CR ²Y is CR ³And Z is CR ⁴

9. the described chemical compound of claim 1, wherein V is CR ⁵, X is CR ²Y is CR ³And Z is CR ⁴

10. the described chemical compound of claim 1, wherein R ^A, R ^A1, R ^BAnd R ^B1Independent separately is H, OH, halogen, C _1-6Alkyl, C _1-6Alkenyl, C _1-6Alkynyl, C _1-6Haloalkyl, C _1-6Alkoxyl or C _1-6Halogenated alkoxy.

11. the described chemical compound of claim 1, wherein R ^A, R ^A1, R ^BAnd R ^B1Independent separately is H, OH or C _1-6Alkoxyl.

12. the described chemical compound of claim 1, wherein R ^A, R ^A1, R ^BAnd R ^B1Independent separately is H or OH.

13. the described chemical compound of claim 1, wherein R ¹Be C _1-6Alkyl, C _1-6Hydroxy alkyl ,-(C _0-6Alkyl)-O-(C _1-6Alkyl) or heterocyclic radical.

14. the described chemical compound of claim 1, wherein R ¹Be C _1-6Alkyl.

15. the described chemical compound of claim 1, wherein R ¹It is third-2-base.

16. the described chemical compound of claim 1, wherein R ⁵And R ⁶One of be not H.

17. the described chemical compound of claim 1, wherein R ⁵And R ⁶One of be C _1-4Haloalkyl.

18. the described chemical compound of claim 1, wherein R ⁶Be C _1-4Haloalkyl.

19. the described chemical compound of claim 1, wherein R ⁶Be CF ₃

20. the described chemical compound of claim 1, wherein R ⁷Be H.

21. the described chemical compound of claim 1, wherein R ⁸Be C _1-3Alkoxyl or C _1-3Halogenated alkoxy.

22. the described chemical compound of claim 1, wherein R ⁸Be C _1-3Alkoxyl.

23. the described chemical compound of claim 1, wherein R ⁸Be methoxyl group.

24. the described chemical compound of claim 1, wherein R ⁸Be ethyoxyl.

25. the described chemical compound of claim 1, wherein R ⁹And R ^{9 '}Be H.

26. the described chemical compound of claim 1 has general formula I a:

Ia。

27. the described chemical compound of claim 1 has general formula I b, Ic or Id:

28. the described chemical compound of claim 1 has general formula I e or If:

29. the described chemical compound of claim 1 has general formula I g:

30. the described chemical compound of claim 1 has general formula I h or Ii:

31. the described chemical compound of claim 1 is selected from:

N-[(1R, 3S)-3-isopropyl-3-(4-[3-(trifluoromethyl) phenyl] and piperazine-1-yl } carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

3-ethyoxyl-N-[(1R, 3S)-3-isopropyl-3-(4-[3-(trifluoromethyl) phenyl] and piperazine-1-yl } carbonyl) cyclopenta] tetrahydrochysene-2H-pyrans-4-amine;

N-[(1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] and piperazine-1-yl } carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

N-[(1R, 3S)-3-isopropyl-3-(4-[5-(trifluoromethyl) pyridin-3-yl] and piperazine-1-yl } carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

N-{ (1R, 3S)-3-isopropyl-3-[(4-phenyl-3,6-dihydropyridine-1 (2H)-yl) carbonyl] cyclopenta }-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

1-((1S, 3R)-1-isopropyl-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino] cyclopenta } carbonyl)-4-Phenylpiperidine-4-alcohol;

1-((1S, 3R)-1-isopropyl-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino] cyclopenta } carbonyl)-4-[2-(trifluoromethyl) phenyl] piperidines-4-alcohol;

1-[((1S, 3R)-1-isopropyl-3-{[3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl] amino } cyclopenta) carbonyl]-4-[3-(trifluoromethyl) phenyl] piperidines-4-alcohol;

1-[((1S, 3R)-1-isopropyl-3-{[3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl] amino } cyclopenta) carbonyl]-4-[4-(trifluoromethyl) phenyl] piperidines-4-alcohol

N-((1R, 3S)-3-isopropyl-3-{[4-[2-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] carbonyl } cyclopenta)-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

N-((1R, 3S)-3-isopropyl-3-{[4-[3-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] carbonyl } cyclopenta)-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

3-ethyoxyl-N-((1R, 3S)-3-isopropyl-3-{[4-[3-(trifluoromethyl) phenyl]-3,6-dihydropyridine-1 (2H)-yl] carbonyl } cyclopenta) tetrahydrochysene-2H-pyrans-4-amine;

N-((1R, 3S)-3-isopropyl-3-{[4-(trifluoromethyl)-3 ', 6 '-dihydro-2,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } cyclopenta)-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

N-((1R, 3S)-3-isopropyl-3-{[5-(trifluoromethyl)-3 ', 6 '-dihydro-3,4 '-bipyridyl-1 ' (2 ' H)-yl] carbonyl } cyclopenta)-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

N-[(1R, 3S)-3-isopropyl-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] and piperazine-1-yl } carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

N-[(1R, 3S)-3-isopropyl-3-(4-[6-(trifluoromethyl) pyridine-2-yl] and piperazine-1-yl } carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

N-[(1R, 3S)-3-isopropyl-3-(4-[6-(trifluoromethyl) pyrimidine-4-yl] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

N-[(1R, 3S)-3-isopropyl-3-(4-[6-methyl-4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

(4R)-N-[(1R, 3S)-3-isopropyl-3-(4-[3-(trifluoromethyl) phenyl] piperidines-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

2-[(1R, 3S)-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino]-1-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] propan-2-ol;

2-[(1R, 3S)-3-[(4R)-3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl] amino-1-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta] propan-2-ol;

2-[(1S, 3S)-3-[(3-methoxyl group tetrahydrochysene-2H-pyrans-4-yl) amino]-1-(4-[6-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] propan-2-ol;

N-[(1S, 3S)-3-ethyl-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

(4R)-N-[(1R, 3S)-3-ethyl-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

N-[(1S, 3S)-3-ethyl-3-(4-[6-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

(4R)-N-[(1R, 3S)-3-methyl-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

(4R)-3-methoxyl group-N-[(1R, 3S)-3-(2-methoxy ethyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] tetrahydrochysene-2H-pyrans-4-amine;

3-methoxyl group-N-[(1S, 3S)-3-(2-methoxy ethyl)-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta] tetrahydrochysene-2H-pyrans-4-amine;

(4R)-N-[(1R, 3S)-3-(ethoxyl methyl)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

(4R)-N-[(1R, 3S)-3-(ethoxyl methyl)-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta]-3-methoxyl group tetrahydrochysene-2H-pyrans-4-amine;

(4R)-3-methoxyl group-N-[(1R, 3S)-3-(methoxy)-3-(4-[4-(trifluoromethyl) pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] tetrahydrochysene-2H-pyrans-4-amine;

(4R)-3-methoxyl group-N-[(1R, 3S)-3-(methoxy)-3-(4-[4-(trifluoromethyl)-pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta] tetrahydrochysene-2H-pyrans-4-amine;

(4R)-3-methoxyl group-N-[(1R, 3S)-3-[(3R)-oxolane-3-yl]-3-(4-[4-(trifluoromethyl)-pyridine-2-yl] piperazine-1-base carbonyl) cyclopenta] tetrahydrochysene-2H-pyrans-4-amine; With

(4R)-3-methoxyl group-N-[(1R, 3S)-3-[(3R)-oxolane-3-yl]-3-(4-[4-(trifluoromethyl) pyrimidine-2-base] piperazine-1-base carbonyl) cyclopenta] tetrahydrochysene-2H-pyrans-4-amine; Or its pharmaceutically acceptable salt.

32. compositions comprises among the claim 1-31 each chemical compound and pharmaceutically acceptable carrier.

33. regulate the method for chemokine receptor activity, comprise making each chemical compound of described chemokine receptors contact claim 1-31.

34. the described method of claim 33, wherein said chemokine receptors are CCR2 or CCR5.

35. the described method of claim 33 wherein saidly is adjusted to inhibition.

36. the described method of claim 33, wherein said chemical compound suppresses CCR2 and CCR5.

37. with the method for chemokine receptor expression or active relevant disease, comprise among the claim 1-31 that gives described patient treatment effective dose each chemical compound among the treatment patient.

38. the described method of claim 37, wherein said chemokine receptors are CCR2 or CCR5.

39. the described method of claim 37, wherein said disease are inflammatory diseases.

40. the described method of claim 37 further comprises giving anti-inflammatory agent.

41. the described method of claim 40, wherein said anti-inflammatory agent are antibody.

42. the described method of claim 37, wherein said disease are dysimmunity.

43. the described method of claim 37, wherein said disease are rheumatoid arthritis, atherosclerosis, lupus, multiple sclerosis, neuropathic pain, transplant rejection, diabetes or obesity.

44. the described method of claim 37, wherein said disease are cancer.

45. the described method of claim 44, wherein said cancer is a feature with the macrophage relevant with tumor.

46. the described method of claim 44, wherein said cancer are breast carcinoma, ovarian cancer or multiple myeloma.

47. the described method of claim 37, wherein said disease or disease are viral infection.

48. being HIV, the described method of claim 47, wherein said viral infection infect.

49. the method that treatment patient HIV infects comprises among the claim 1-31 that gives described patient treatment effective dose each chemical compound.

50. the described method of claim 49 further comprises the while or gives at least a antiviral agents successively.