CN101253168B

CN101253168B - Substituted benzimidazoles as kinase inhibitors

Info

Publication number: CN101253168B
Application number: CN200680031748.1A
Authority: CN
Inventors: M·E·艾卡娃; P·阿米里; J·H·达夫; B·H·莱文; C·麦克布赖德; T·E·皮克; D·J·蓬; S·拉默西; P·A·伦豪威; C·谢弗; D·斯图尔特; S·苏布拉马尼安
Original assignee: Novartis AG
Current assignee: Novartis AG
Priority date: 2005-08-30
Filing date: 2006-08-30
Publication date: 2012-12-12
Anticipated expiration: 2026-08-30
Also published as: ZA200800506B; ES2360608T3; CN101253168A; UA94055C2

Abstract

New substituted benzimidazole compounds, compositions, and methods of inhibition of kinase activity associated with tumorigenesis in a human or animal subject are provided. In certain embodiments, the compounds and compositions are effective to inhibit the activity of at least one serine/threonine kinase or receptor tyrosine kinase. The new compounds and compositions may be used either alone or in combination with at least one additional agent for the treatment of a serine/threonine kinase- or receptor tyrosine kinase-mediated disorder, such as cancer.

Description

Substituted benzimidazoles as SU11752

The cross reference of related application

It is 60/712 that the application requires in the provisional application sequence number of application on August 30th, 2005; 539 U.S. Patent application, the provisional application sequence number of applying on August 30th, 2005 are 60/713; 108 U.S. Patent application, the provisional application sequence number of applying on October 27th, 2005 are 60/731; 591 U.S. Patent application and the provisional application sequence number of applying on February 17th, 2006 are 60/774; The right of priority of 684 U.S. Patent application, these patented claims here all are incorporated herein by reference in full.

Technical field

The application relates to new substituted benzimidazole compound, its tautomer, steric isomer, polymorphic form, ester, metabolite and prodrug; The pharmaceutically useful salt of said compound, tautomer, steric isomer, polymorphic form, ester, metabolite and prodrug; The compsn of above-mentioned any embodiment and pharmaceutically acceptable carrier, and above-mentioned any embodiment is united the application in prevention or treatment cancer separately or with at least a other therapeutical agent.

Background technology

Known and tumour generate relevant kinases and comprise Raf serine/threonine kinase and receptor tyrosine kinase (RTK).

This Raf serine/threonine kinase is the essential component of control to Ras/ mitogen-activated protein kinase (MAPK) signal transduction module of the complicated transducer of outside cytositimulation generation.Known serine-threonine-the specificity protein kinase guarded with ras oncogene bonded height of Raf genes encoding.It is an a kind of part of signal transduction pathway, it is believed that this signal transduction pathway is made up of its final phosphorylation transcription factor receptor tyrosine kinase, p21 ras, Raf protein kinase, Mek1 (ERK acvator or MAPKK) kinases and ERK (MAPK) kinases.In this approach, the Raf kinases is by two kinds of hypotypes of Ras activation and phosphorylation and activation mitogen-activated protein kinase kinases (so-called Mek1 and Mek2), and it is the Threonine/Tyrosylprotein kinase of dual specificity.Two kinds of Mek hypotypes all activation mitogen activatory kinases 1 and 2 (MAPK, also be called as kinases 1 that extracellular ligand regulates and 2 or Erk1 and Erk2).This MAPK phosphorylation comprises many substrates of transcription factor and sets up its transducer thus.Participate in the Raf kinases influence of Ras/MAPK approach and regulating many cell functions such as propagation, differentiation, survival, carinogenicity conversion and apoptosis.

In mammalian cell, use use a model biological biological chemistry and the research of genetic technique of research and some of the Raf mutant that deregulated and dominance suppresses to prove vital role and the position of Raf in many signal transduction pathways by some.In many cases, the acceptor of irritation cell tyrosine phosphorylation depends on the activity of Ras to the activation of Raf, and it shows that Ras is in the upper reaches of Raf performance function.When activation, Raf-1 phosphorylation and activation Mek1, thus make signal be transmitted to the downstream effect device, like MAPK (mitogen-activated protein kinase; People such as Crews, 1993, Cell 74:215).Think that said Raf serine/threonine kinase is main Ras effector related in the zooblast propagation (people such as Avruch, 1994, Trends Biochem.Sci.19:279).

The Raf kinases has three kinds of different hypotypes---Raf-1 (c-Raf), A-Raf and B-Raf; Its difference is ability, tissue distribution and the Subcellular Localization (people such as Marias of itself and the interactional ability of Ras, activation mapk kinase approach; Biochem.J.351:289-305,2000; People such as Weber, oncogene 19:169-176,2000; People such as Pritchard, Mol.Cell.Biol.75:6430-6442,1995).In whole tumours about 20% in can find a kind of activation sudden change of Ras gene and in whole tumours about 30% described in the Ras/Raf/MEK/ERK approach be activated (people such as Bos, Cancer Res.49:4682-4689,1989; People such as Hoshino, Oncogene18:813-822,1999).Some nearest researchs have shown that the B-Raf sudden change in the cutaneous nevus is the committed step that begins of melanophore tumorigenesis people such as (, Nature Genetics 25:1-2,2002) Pollock.In addition, up-to-date some research discloses activation sudden change that the B-Raf kinase domain has taken place in the liver cancer of melanoma about 66%, 12% colorectal carcinoma and 14% (people such as Davies, Nature417:949-954,2002; People such as Yuen, Cancer Research 62:6451-6455,2002; People such as Brose, Cancer Research 62:6997-7000,2002).

Melanoma is a medical problem of failing fine solution always, and it is a kind of multigenic disease of poor prognosis of complicacy, and is especially true in the transfering state late.In many malignant tumours, found the active body cell mutation in the B-Raf proto-oncogene recently, and the most common in melanoma.About 70% melanoma is expressed variation and the activatory form of B-Raf (V600E), and this makes it become a kind of target spot of outstanding drug development.In addition, the melanoma of 10-15% is expressed mutant N-Ras in addition, and it has proved that further the MAPK approach is in the growth of melanoma cells and the importance in the survival.

Under Raf kinases level, the suppressor factor of Ras/Raf/MEK/ERK approach possibly be antagonism had express or receptor tyrosine kinase, the kinase whose tumour of activatory intracellular tyrosine of variation, have unconventionality expression Grb2 (a kind of making can stimulate the adaptin of Ras through the Sos exchange factor) tumour and conceal the tumor treatment agent that the activity of Raf itself is suddenlyd change.In the clinical trial, shown that the kinase whose suppressor factor of the Raf-1 that also suppresses B-Raf is promising therapeutical agent (Crump, Current Pharmaceutical Design 8:2243-2248,2002 in treatment for cancer in early days; People such as Sebastien, Current Pharmaceutical Design 8:2249-2253,2002).

Shown through using RNA antisense technology destroyed Raf expression inhibiting in the clone tumorigenicity that mediates of Ras and Raf-(people such as Kolch, Nature 349:416-428,1991; People such as Monia, Nature Medicine 2 (6): 668-675,1996).Shown that also the deactivation antibody---the kinase whose substrate of Raf---that resists the Raf kinases or bear the coexpression of dominance Raf kinases or negative dominance MEK can make cell transformed demobilize into normal growth phenotype and (see people such as Daum; TrendsBiochem.Sci 1994,19:474-80; People J Biol.Chem.1994 such as Fridman, 269:30105-8).

Some Raf SU11752 have been described in external and/or in vivo tests, effectively to suppress tumor cell proliferation (referring to for example US patent 6,391,636,6,358,932,6; 037,136,5,717,100,6,458; 813,6,204,467 and 6,268,391).Other patents and patented claim show that the Raf SU11752 can be used for treating white blood disease (referring to for example US patent 6,268,391 and 6,204,467 and disclosed US patented claim 20020137774; 20020082192; 20010016194; With 20010006975) or be used to treat breast cancer (referring to for example US patent 6,358,932,5,717,100,6,458,813,6,268,391 and 6,204,467 and disclosed US patented claim 20010014679).

Blood vessel takes place also in growth of cancer cells, to play an important role.In case the place of hiding of known cancer cells reaches certain size, when approximately diameter was 1 to 2mm, because the diffusion deficiency thinks that cancer cells provides competent oxygen and nutrition, tumor growth got bigger so cancer cells must be set up supply of blood flow.Therefore, but expection suppresses the growth that anticancer takes place blood vessel.

Receptor tyrosine kinase (RTK) is that reconstruct and the regenerated of regulating developmental cell growth and differentiation, mature tissue are striden membrane polypeptides (Mustonen, people such as T., J Cell Biology 129:895-898,1995; Van der Geer, people such as P., Ann Rev.Cell Biol.10:251-337,1994).But known some polypeptide ligand activation RTK that are called as growth factor or cytokine.The RTK signal transduction relates to the transfer (Lymboussaki that part combines and cause the acceptor external structure territory conformation of its dimerisation; A. " VEGF in embryo, adult and the tumour with and acceptor (VascularEndothelial Growth Faxtors and their Receptors in Embryos; Adults; andin Tumors) ", Academic Dissertation, University of Helsinki; Molecular/Cancer Biology Laboratory and Department of Pathology.Haartman Institute, 1999; Ullrich, people such as A., Cell 61:203-212,1990).This part and RTK are combined in the activation that has caused the acceptor transphosphorylation on the specificity tyrosine residues and caused the required catalyst structure domain of kytoplasm substrate (Id) phosphorylation subsequently.

Two subtribes of RTK have specificity to blood vessel endothelium.These subtribes comprise VEGF (VEGF) subtribe and Tie acceptor subtribe.V class RTK comprises VEGFR1 (FLT-I), VEGFR2 (KDR (people), FIk-1 (mouse)) and VEGFR3 (FLT-4) (Shibuya, people such as M., Oncogene5:519-525,1990; Terman, people such as B., Oncogene 6:1677-1683,1991; Aprelikova, people such as O., Cancer Res.52:746-748,1992).The member of VEGF subtribe be described to can the induction of vascular perviousness with endothelial cell proliferation and further be accredited as the main inductor (Ferrara, people such as N., Endocrinol.Rev.18:4-25,1997) of blood vessel generation and vasculogenesis.

Known VEGF combines (DeVries, people such as C., Science 255:989-991,1992 with the RTK specificity that comprises FLT-1 and FIk-1; Quinn, people such as T., Proc.Natl.Acad.Sci.90:7533-7537,1993).(Connolly, people such as D., J.Biol.Chem.264:20017-20024,1989 take place with the migration of external stimulating endothelial cells and propagation and induction of vascular in VEGF in vivo; Connolly, people such as D., J.Clin.Invest.84:1470-1478,1989; Ferrara, people such as N., Endocrinol.Rev.18:4-25,1997; Leung, people such as D., Science 246:1306-1309,1989; Plouet, people such as J., EMBO J 5:3801-3806,1989).

The research of in various endothelial cells cultured system, carrying out has confirmed that VEGFR2 has mediated the most downstream effects of VEGF in blood vessel takes place (people such as Wey S.; ClinicalAdvances in Hematology and Oncology; 2:37-45,2004).Think the endothelial cell proliferation that in the activation of Ras/Raf/Mek/Erk approach, relates to VEGFR2 mediation people such as (, Cancer Res 60:203-212,2000) Veikkola T..In melanoma, breast cancer, bladder cancer, lung cancer, thyroid carcinoma, prostate cancer and ovarian cancer, all observed the expression (seeing people such as Wey, the same) of VEGFR2.The neutralizing monoclonal antibody (KDR) that has shown VEGFR2 can effectively be blocked tumor vessel and take place (to see people such as Kim, Nature 362:841,1993; People such as Rockwell, Mol.Cell Differ.3:315,1995).Because it is very crucial and by VEGF and VEGF-RTK control for the growth of cancer that known blood vessel takes place, has carried out many effort and developed and to suppress or line artery takes place and the compound of inhibition VEGF-RTK.

Platelet derived growth factor receptor kinases (PDGFR) is the another kind of type of RTK.Being illustrated in many different solid tumors (from glioblastoma and osteosarcoma to prostate cancer) all has PDGF to express.In these various types of tumours, the biological action of PDGF signal can interact from autocrine stimulation to the more delicate paracrine that relates to the generation of adjacent matrix and blood vessel of growth of cancer cells.PDGF and tyrosine kinase receptor PDGFR α and PDGFR β interact.Therefore, expection can disturb tumor growth and blood vessel to take place with small molecules inhibition PDGFR kinase activity.

Fibroblast growth factor acceptor kinases (FGFR) has been represented the another kind of type of RTK.This fibroblast growth factor is the PGF family that participates in comings and goings, and said activity comprises that mitogenesis, blood vessel take place and wound healing.It comprises the relevant still different separately tyrosine kinase receptor of gang, and said acceptor comprises an ectodomain with 2 or 3 Tegelines (Ig)-spline structure territory, membrane spaning domain and cytoplasmic tyrosine kinase structural domain.The fibroblast growth factor acceptor of having confirmed comprises FGFR1 (Ruta, people such as M, Oncogene 3:9-15,1988); FGFR2 (Dionne, people such as C, Cytogenet.Cell Genet.60:34-36,1992); FGFR3 (Keegan, people such as K, Proc.Nat.Acad.Sci.88:1095-1099,1991); And FGFR4 (Partanen, people such as J, EMBO J.10:1347-1354,1991).

The effect in cancer of fibroblast growth factor acceptor, particularly FGFR3 has been described.The imbalance of the oncogene of carrying out through the heavy chain immunoglobulin on 14q32 (IgH) position transposition is the basic incident in the pathogeny of B-glucagonoma.In multiple myeloma, the transposition to the IgH position has taken place in 20 to 60% case.For most of transposition, its pairing chromosome is unknown; For some other situation, confirmed the to have 11q13 different dyeing body of (unique karyomit(e) that is usually related to) is to arranging.People such as Bergsagel are accredited as irrational conversion recombinant fragment (be defined as to comprise and derive from the only sequence of 1 transition region) the possible affinity tag of the transposition incident of the IgH transition region among 15 of 21 myeloma cell lines, comprise 7 in being of 8 caryogram (karyotyped) with the 14q32 transposition that can't detect.These transposition breakpoints relate to 6 chromosome region: 4pl6.3; 6; 8q24.13; 11q13.3; 16q23.1; And 21q22.1 (people such as Bergsagel, Proc.Nat.Acad.Sci.93:13931-13936,1996).People such as Chesi (Nature Genet.16:260-264 1997) find in 5 kinds of myeloma cell lines and the reticent transposition t (4 of caryogram in 10 kinds of with the primary tumor of multiple myeloma 3 kinds at least; 14) (p16.3; Q32.3) show expression and the activation increase that FGFR3 suddenlys change.Said karyomit(e)-4 breakpoint accumulates in the kinetochore to the 70-kb zone of FGFR3, thinks that it is to regulate bad (dysregulated) oncogene.Two kinds of clones and a kind of primary tumor selective expression with this transposition comprise the FGFR3 allelotrope of definite activation sudden change (tyr373 to cys, lys650 to glu and lys650 to met) in thanatophoric dwarfism before.For K650E, proved do not have the composition activation of FGFR3 under the part situation with transfection experiment.People such as Chesi (1997) propose at said t (4; 14) after the transposition, the somatic mutation during the tumor development usually is created in and does not have activated FGFR3 albumen under the part situation.

Rasmussen, people such as T propose t (4 in multiple myeloma; 14) frequency of transposition is 3 to 24% (Rasmussen, people such as T, Br.J.Haematol.117:626-628,2002).The frequency of in the patient who suffers from the not bright mono-clonal Agammaglobulinemia (MGUS) of meaning, observing this transposition is significantly lower, and this has shown that it is from the effect of MGUS to the transformation of multiple myeloma.Said t (4; 14) be prone to 2 potential oncogene: FGFR3 of position influence and multiple myeloma SET structural domain (MMSET).People such as Rasmussen (2002) to the frequency of FGFR3 imbalance in the multiple myeloma with and prognosis values study.Its FGFR3 that in 16 (14.5%) of 110 multiple myeloma marrow samples, has found imbalance expresses.

In addition, there has been the other sign to show the carcinogenesis (Cappellen, people such as D., (Letter) Nature Genet.23:18-20,1999) of FGFR3 in cancer.People such as Cappellen are in 2 kinds of common epithelial cancers of significant proportion---found to form the expression of activatory FGFR3 in bladder and the cervical cancer.FGFR3 is the most frequent oncogene of variation in the bladder cancer seemingly, and it morphs in the case more than 30%.As if FGFR3 mediated opposite signal, in osteogenesis the performance negativity modulator effect and in the tumour of some types as oncogene.All FGFR3 missense somatic mutations of in these cancers, confirming are equal to the embryo property activation sudden change that causes thanatophoric dysplasia, and (author notices in 2 kinds of sudden changes; It takes place equably, and this is because the FGFR3b hypotype in epithelial cell, expressed is Duoed 2 amino acid than the FGFR3c hypotype of in bone, expressing).In the FGFR3 of epithelial tumor changed, modal was the S249C sudden change, and it affects 3 examples in example of 5 in the 9 routine bladder cancer and the 3 routine cervical cancers.

Also have sign to show that activatory FGFR3 is the target spot of degrading through the lysosome that the ubiquitination that c-Cbl-mediates carries out; And be illustrated in the activation sudden change of finding in patient's body of suffering from fetal rickets and relevant dyschondroplasia and upset this process; Caused the recycling of activated receptor and the amplification of FGFR3 signal (people such as Cho; Proc.Nat.Acad.Sci.101:609-614,2004).People such as Cho propose the potential target spot that this mechanism helps achondroplastic molecular pathogenesis and represented treatment to intervene.This lysosome target defective has been explained achondroplastic pathogeny with other mechanism that is proposed.

Some other result shows that FGFR2 and FGFR3 are important factor (people such as Jang JH in tumour generates; " with the sudden change (Mutations in fibroblastgrowth factor 2 and fibroblast growth factor 3 genes associated withhuman gastric and colorectal cancers) in people's cancer of the stomach and Cancer-Related fibroblast growth factor acceptor 2 of colorectum and the fibroblast growth factor receptor3 gene "; Cancer Res.61 (9): 3541-3,2001).Because its effect in multiple myeloma, bladder cancer and tumour generate, the suppressor factor that is developed to bfgf receptor kinase whose suppressor factor, particularly FGFR2 and FGFR3 will play an important role in treatment for cancer.

C-Kit is that another kind belongs to the receptor tyrosine kinase of pdgf receptor family and normal expression in HPC, mastocyte and sexual cell.In many cancers, all relating to C-kit expresses; These cancers comprise mast cell leukemia, germinoma, small cell lung cancer, gastrointestinal stromal tumor, acute myelogenous leukemia (AML), erythroleukemia, neuroblastoma, melanoma, ovarian cancer, breast cancer (Heinrieh, people such as M.C.; J.Clin.Onc.20,61692-1703,2002 (survey articles); Smolich, people such as B.D., Blood, 97,5; 1413-1421).

In many human cancers, relating to crossing of CSF-1R (acceptor of colony-stimulating factor-1 (CSF-1)) expresses; Said cancer comprises breast cancer, ovarian cancer, carcinoma of endometrium, lung cancer, kidney, carcinoma of the pancreas and prostate cancer (Sapi; E., Exp.Biol.Med 229:1-11,2004).CSF-1R is a tyrosine kinase receptor, and when by its part CSF-1 activation, it has triggered the signal transduction pathway of control cell proliferation and differentiation.In pregnancy with between lactation, CSF-1R expresses in mammary gland.Show, unusual CSF-1R express with all breast cancers in 58% relevant, and with the wellability breast cancer in 85% relevant (seeing Sapi, the same).

Need to suppress blood capillary proliferation always, suppress tumor growth, can treat cancer, cell cycle regulation is stagnated and/or suppress molecule such as Ras, Raf, mutant B-Raf, VEGFR2 (KDR, Flk-1), one or more compound and pharmaceutical prepn and the medicine that comprises this compounds among the FGFR2/3, c-Kit, PDGFR β, CSF-1R.Also need some this compounds, pharmaceutical prepn and medicine to be delivered medicine to patient or the individual method that needs it.

General introduction of the present invention

The invention provides the novel substituted benzimidazole compound of formula (I) or the pharmaceutically useful salt of its tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug or this compound, tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug:

Wherein,

Each R ¹Be independently selected from hydroxyl, halogen, C _1-6Alkyl, C _1-6Alkoxyl group, (C _1-6Alkyl) sulfenyl, (C _1-6Alkyl) alkylsulfonyl, naphthenic base, Heterocyclylalkyl, phenyl and heteroaryl;

R ²Be C _1-6Alkyl or halo (C _1-6Alkyl);

Each R ³Be independently selected from halogen, C _1-6Alkyl and C _1-6Alkoxyl group;

Each R ⁴Be independently selected from hydroxyl, C _1-6Alkyl, C _1-6Alkoxyl group, halogen, Heterocyclylalkyl carbonyl, carboxyl, (C _1-6Alkoxyl group) carbonyl, aminocarboxyl, C _1-6Alkyl amino-carbonyl, cyanic acid, naphthenic base, Heterocyclylalkyl, phenyl and heteroaryl;

R wherein ¹, R ², R ³And R ⁴Can be randomly by one or more hydroxyl, halogen, C of being independently selected from _1-6Alkyl, halo (C _1-6Alkyl), C _1-6Alkoxyl group and halo (C _1-6Alkoxyl group) substituting group replaces;

A is 1,2,3,4 or 5;

B is 0,1,2 or 3; And

C is 1 or 2.

In some other embodiment, the novel substituted benzimidazole compound of formula (II) or the pharmaceutically useful salt of its tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug or this compound, tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug are provided:

Wherein,

Each R ¹Be independently selected from C _1-6Alkyl, C _1-6Alkoxyl group, hydroxyl, halogen, (C _1-6Alkyl) sulfenyl, (C _1-6Alkyl) alkylsulfonyl, naphthenic base, Heterocyclylalkyl, phenyl and heteroaryl;

Each R ⁴Be independently selected from hydroxyl, C _1-6Alkyl, C _1-6Alkoxyl group, halogen, carboxyl, (C _1-6Alkoxyl group) carbonyl, aminocarboxyl, cyanic acid, naphthenic base, Heterocyclylalkyl, Heterocyclylalkyl carbonyl, phenyl and heteroaryl;

R wherein ¹, R ², R ³And R ⁴Can be randomly by one or more hydroxyl, halogen, C of being independently selected from _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces;

A is 1,2,3,4 or 5;

B is 0,1,2 or 3; And

C is 1 or 2.

In some other embodiment, the novel substituted benzimidazole compound of formula (III) or the pharmaceutically useful salt of its tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug or this compound, tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug are provided:

Wherein,

R wherein ¹And R ⁴Can be randomly by one or more hydroxyl, halogen, C of being independently selected from _1-6Alkyl and C _1-6The substituting group of alkoxyl group replaces;

A is 1,2,3,4 or 5; And

C is 1 or 2.

The compound of following formula (IV) or the pharmaceutically useful salt of its tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug or this compound, tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug are also disclosed:

Wherein,

R ²Be C _1-6Alkyl or halo (C _1-6Alkyl);

Each R ⁴Be independently selected from hydroxyl, C _1-6Alkyl, C _1-6Alkoxyl group, halogen, carboxyl, (C _1-6Alkoxyl group) carbonyl, aminocarboxyl, C _1-6Alkyl amino-carbonyl, cyanic acid, cyanic acid (C _1-6Alkyl), naphthenic base, Heterocyclylalkyl, Heterocyclylalkyl (C _1-6Alkyl), Heterocyclylalkyl carbonyl, phenyl and heteroaryl;

A is 1,2,3,4 or 5; And

B is 0,1,2 or 3.

In some other embodiment, the novel substituted benzimidazole compound of formula (I)-(IV) is provided, wherein each R ¹Be independently selected from hydroxyl, chlorine, fluorine, bromine, methyl, ethyl, propyl group, butyl, methoxyl group, oxyethyl group, propoxy-, butoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoro ethoxy, trifluoromethylthio, piperidyl, C _1-6Alkylpiperidine base, piperazinyl, C _1-6Alkylpiperazine base, tetrahydrofuran base, pyridyl and pyrimidyl.In some other embodiment, the novel substituted benzimidazole compound of formula (I)-(IV) is provided, wherein a is 1 or 2, and at least one R ¹Be halo (C _1-6Alkyl), like trifluoromethyl.In some other embodiment, formula (I) and novel substituted benzimidazole compound (IV), wherein R are provided ²Be C _1-6Alkyl, as, for example, methyl or ethyl.In other embodiments, formula (I), (II) and novel substituted benzimidazole compound (IV) are provided, wherein b is 0, and so R ³Do not exist.In other embodiments, the novel substituted benzimidazole compound of formula (I)-(IV) is provided, wherein b is 1, and R ³Be C _1-6Alkoxyl group, as, for example, methoxyl group.In other embodiments, the novel substituted benzimidazole compound of formula (I)-(III) is provided, wherein c is 1 or 2, and at least one R ⁴Be halo (C _1-6Alkyl), as, for example, trifluoromethyl.

The method of the illness relevant with Raf that the human or animal that the present invention provides treatment to need such treatment on the other hand is individual, it comprises formula (I), (II), (III) or compound (IV) or its pharmaceutically useful salt of using any embodiment that can effectively reduce or prevent this individuality tumor growth in vivo quantity to said individuality.

The method of the illness relevant with Raf that the human or animal that the present invention also provides treatment to need such treatment on the other hand is individual, it comprises to said individuality uses formula (I), (II), (III) or compound (IV) or its pharmaceutically useful salt of any embodiment that can effectively reduce or prevent this individuality tumor growth in vivo quantity and unites at least a other promoting agent that is used to treat cancer of use.

The present invention also provides on the other hand and has comprised the therapeutic compsn at cancer therapy other promoting agent commonly used that at least a formula (I), (II), (III) or compound (IV) or its pharmaceutically useful salt and one or more are used for treating cancer.

Compound of the present invention can be used for treating cancer; Comprise that cancer (for example; Lung, pancreas, ovary, Tiroidina, bladder or colorectal carcinoma), melanoma, marrow illness (for example; Myelocytic leukemia, multiple myeloma and erythroleukemia), adenoma (for example, fine hair shape adenoma of colon) and sarcoma (for example, osteosarcoma).

On the other hand; The present invention relates to suppress in the individual MAPK signal transduction pathway at least a serine/threonine kinase or treatment is by the method for the biology situation of the mediation of the serine/threonine kinase in the MAPK signal transduction pathway in individuality, it comprises using and comprises at least a therapeutic compsn that can effectively suppress formula (I), (II), (III) or the compound or pharmaceutically acceptable salt thereof (IV) of individual intravital MAPK signal transduction pathway.Said therapeutic compsn can be used for treating the patient's (for example, suffer from by the unusual cancers mediated of MAPK signal transduction patient) who such suppressor factor is had demand.In one embodiment; The present invention relates to suppress the kinase whose method of individual intravital Raf, it comprise use comprise { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine with and the therapeutic compsn of the pharmaceutically useful salt of tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug or this compound, tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug.

On the other hand; The present invention relates to suppress the tyrosine kinase receptor of at least a VEGFR-2 of being selected from the individual's body, PDGFR-β, pERK, bFGF, FGFR1, FGFR2, FGFR3, c-Kit and CSF-1R or treat the method by the biology situation of at least a mediation among VEGFR-2, PDGFR-β, pERK, bFGF, FGFR1, FGFR2, FGFR3, c-Kit and the CSF-1R, it comprises uses the therapeutic compsn that comprises at least a formula (I), (II), (III) or the compound or pharmaceutically acceptable salt thereof (IV) that can effectively suppress individual intravital tyrosine kinase receptor.This treatment compound can be used for treating the patient's (for example, suffer from by the unusual cancers mediated of tyrosine kinase receptor signal transduction patient) who such suppressor factor is had demand.In one embodiment; The present invention relates to suppress be selected from the individual's body method of the Tyrosylprotein kinase of VEGFR-2, PDGFR-β, pERK, bFGF, FGFR1, FGFR2, FGFR3, c-Kit and CSF-1R, it comprises the therapeutic compsn of using the pharmaceutically useful salt that comprises { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine or its tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug or this compound, tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug.

The present invention also provides compsn, application method and the working method described in the present invention's detailed description.

Brief Description Of Drawings

In conjunction with accompanying drawing, through with reference to following detailed, above-mentioned aspect of the present invention and many advantages that accompanies will be easier to understand, wherein:

Fig. 1 be expression when as embodiment 78 described, when treating with compound of the present invention, the figure of the average reduction of the interior A375M human melanoma gross tumor volume of mouse body;

Fig. 2 A and 2B are some PAGE figure; Its expression is as described in the embodiment 79; Treating back 4-(Fig. 2 A) and 24-hour (Fig. 2 B) with compound of the present invention, to the inhibition of the kinase whose downstream signal transduction of Raf in the A375M human melanoma tumour cell in the mouse body;

Fig. 3 be expression when as embodiment 80 described, when treating with compound of the present invention, the figure of the average reduction of the interior HT29P human colon carcinoma gross tumor volume of mouse body;

Fig. 4 A, 4B and 4C are some PAGE figure; Its expression is as described in the embodiment 81; 1 hour (Fig. 4 A), 4 hours (Fig. 4 B) and 24 hours (Fig. 4 C) after with compounds for treating of the present invention are to the inhibition of the downstream signal transduction of HT29P human colon carcinoma tumour cell in the mouse body;

Fig. 5 has explained MAPK signal transduction pathway that comprises Ras, Raf, MEK and ERK and the inhibition point that as embodiment 82-86 is described, suppresses the kinase whose downstream signal of Raf with the compound of embodiment 1 of being advised;

Fig. 6 A, 6B and 6C are some PAGE figure; Its expression is as described in the embodiment 83; After in culture, cultivating 4 hours with the compound of the embodiment 1 of finite concentration scope, to the inhibition of the kinase whose downstream signal of Raf in A375M cell (Fig. 6 A), SK-MEL2 cell (Fig. 6 B) and the CHL-1 cell (Fig. 6 C);

Fig. 7 A is that expression is when as embodiment 84 is described; When treating with the oral dosage of 10mg/kg, 30mg/kg or 100mg/kg embodiment 1 compound, the figure of the dose response that A375M (B-Raf V600E) human melanoma gross tumor volume on average reduces in the mouse body;

Fig. 7 B is PAGE figure, and it is illustrated in after embodiment 84 described such compounds with embodiment 1 carry out treating 8 hours the 14th time, to the inhibition of the kinase whose downstream signal of Raf in the A375M tumour cell in the mouse body;

Fig. 7 C is PAGE figure, and it is illustrated in after embodiment 84 described such compounds with embodiment 1 carry out treating 24 hours the 14th time, to the inhibition of the kinase whose downstream signal of Raf in the A375M tumour cell in the mouse body;

Fig. 7 D is PAGE figure, and it is illustrated in after embodiment 84 described such compounds with embodiment 1 carry out treating 24 hours the 14th time, to the regulation and control of the affinity tag in Raf kinases downstream in the A375M tumour cell;

Fig. 8 A is expression when as embodiment 85 is described, treating with the compound of embodiment 1, the figure of the average reduction of interior MEXF276 (B-Raf V600E) the melanoma cancerous tumour volume of mouse body;

Fig. 8 B is PAGE figure, and it is illustrated in after embodiment 85 described such compounds with embodiment 1 are treated 4 hours the 20th time, to the inhibition of the kinase whose downstream signal of Raf in the MEXF276 tumour cell in the mouse body;

Fig. 8 C is PAGE figure, and it is illustrated in after embodiment 85 described such compounds with embodiment 1 are treated 4 hours the 20th time, to the regulation and control of the affinity tag in Raf kinases downstream in the MEXF276 tumour cell;

Fig. 9 A is expression when as embodiment 85 is described, treating with the compound of embodiment 1, the figure of the average inhibition that MEXF1341 in the mouse body (N-Ras Q61K) melanoma cancerous tumour is grown;

Fig. 9 B is PAGE figure, and it is illustrated in after embodiment 85 described such compounds with embodiment 1 carry out treating 4 hours the 20th time, the kinase whose downstream signal of Raf in the interior MEXF1341 tumour cell of mouse body;

Fig. 9 C is PAGE figure, and it is illustrated in after embodiment 85 described such compounds with embodiment 1 are treated 4 hours the 20th time, to the regulation and control of the affinity tag in Raf kinases downstream in the MEXF1341 tumour cell;

Figure 10 A is expression when as embodiment 86 is described, treating with the compound of embodiment 1, the figure of the average reduction of interior HCT-116 (K-Ras G13D) the colorectal carcinoma gross tumor volume of mouse body;

Figure 10 B is PAGE figure, and it is illustrated in after embodiment 86 described such compounds with embodiment 1 carry out treating 4 hours the 3rd time, to the inhibition of the kinase whose downstream signal of Raf in the HCT-116 tumour cell in the mouse body;

Figure 10 C is PAGE figure, and it is illustrated in after embodiment 86 described such compounds with embodiment 1 carry out treating 8 hours the 3rd time, to the inhibition of the kinase whose downstream signal of Raf in the HCT-116 tumour cell in the mouse body;

Figure 10 D is PAGE figure, and it is illustrated in after embodiment 86 described such compounds with embodiment 1 carry out treating 24 hours the 3rd time, to the inhibition of the kinase whose downstream signal of Raf in the HCT-116 tumour cell in the mouse body;

Figure 11 is expression when as embodiment 86 is described, treating with the compound of embodiment 1, the figure of the average reduction of interior HT-29 (B-Raf V600E) the colorectal carcinoma gross tumor volume of mouse body;

Figure 12 A is expression when as embodiment 86 is described, treating with the compound of embodiment 1, the figure of the average inhibition that MV4-11 in the mouse body (FLT3 ITD) acute monocytic leukemia cancerous tumour is grown;

Figure 12 B is PAGE figure, and it is illustrated in after embodiment 86 described such compounds with embodiment 1 carry out treating 4 hours the 3rd time, the kinase whose downstream signal of Raf in the interior MV4-11 tumour cell of mouse body;

Figure 13 be illustrated in as embodiment 88 is described treat with the compound of 10mg/kg, 30mg/kg and 100mg/kg embodiment 1 after, the figure of the inhibition that the blood vessel of VEGF-mediation in the CHO-VEGF Matrigel model is taken place;

Figure 14 A is expression when as embodiment 89 is described, treating with the dosage regimen of q2d, q3d or q4d with the compound of 100mg/kg embodiment 1, the figure of the average reduction of the interior A375M melanoma gross tumor volume of mouse body;

Figure 14 B is PAGE figure; It is illustrated in, and the compound with embodiment 1 carries out the 5th treatment after 8 hours, 24 hours and 48 hours, to the inhibition of the kinase whose downstream signal of Raf in the A375M tumour cell in the mouse body with the dosage regimen of q2d as embodiment 89 is described;

Figure 14 C is PAGE figure; It is illustrated in, and the compound with embodiment 1 carries out the 3rd treatment after 48 hours, 72 hours and 96 hours, to the inhibition of the kinase whose downstream signal of Raf in the A375M tumour cell in the mouse body with the dosage regimen of q4d as embodiment 89 is described; With

Figure 15 be expression as described in the embodiment 90, the figure that the A375M tumour cell is treated, concerned between the concentration threshold of serum-concentration and the target adjusting of this compound as time goes by with the compound of various concentration embodiment 1.

Detailed description of the preferred embodiments

According to an aspect of the present invention, the substituted benzimidazole compound of formula (I) or the pharmaceutically useful salt of its tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug or this compound, tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug are provided:

Wherein,

R ²Be C _1-6Alkyl or halo (C _1-6Alkyl);

Each R ⁴Be independently selected from hydroxyl, C _1-6Alkyl, C _1-6Alkoxyl group, halogen, carboxyl, (C _1-6Alkoxyl group) carbonyl, aminocarboxyl, C _1-6Alkyl amino-carbonyl, cyanic acid, naphthenic base, Heterocyclylalkyl, Heterocyclylalkyl carbonyl, phenyl and heteroaryl;

A is 1,2,3,4 or 5;

B is 0,1,2 or 3; And

C is 1 or 2.

Wherein,

A is 1,2,3,4 or 5;

B is 0,1,2 or 3; And

C is 1 or 2.

Wherein,

A is 1,2,3 or 5; And

C is 1 or 2.

Wherein,

R ²Be C _1-6Alkyl or halo (C _1-6Alkyl);

A is 1,2,3,4 or 5; And

B is 0,1,2 or 3.

In some other embodiment, the novel substituted benzimidazole compound of formula (I)-(IV) is provided, wherein each R ¹Be independently selected from hydroxyl, chlorine, fluorine, bromine, methyl, ethyl, propyl group, butyl, methoxyl group, oxyethyl group, propoxy-, butoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoro ethoxy, trifluoromethylthio, piperidyl, C _1-6Alkylpiperidine base, piperazinyl, C _1-6Alkylpiperazine base, tetrahydrofuran base, pyridyl and pyrimidyl.In some other embodiment, the novel substituted benzimidazole compound of formula (I)-(IV) is provided, wherein a is 1 or 2, and at least one R ¹Be halo (C _1-6Alkyl), like trifluoromethyl.In some other embodiment, formula (I) and novel substituted benzimidazole compound (IV), wherein R are provided ²Be C _1-6Alkyl, as, for example, methyl or ethyl.In other embodiments, formula (I), (II) and novel substituted benzimidazole compound (FV) are provided, wherein b is 0, and so R ³Do not exist.In other embodiments, the novel substituted benzimidazole compound of formula (I)-(IV) is provided, wherein b is 1, and R ³Be C _1-6Alkoxyl group, as, for example, methoxyl group.In other embodiments, the novel substituted benzimidazole compound of formula (I)-(III) is provided, wherein c is 1 or 2, and at least a R ⁴Be halo (C _1-6Alkyl), as, for example, trifluoromethyl.

In some embodiments, R ¹, R ², R ³And R ⁴Can randomly be independently selected from hydroxyl, halogen, C by one to five _1-6Alkyl, halo (C _1-6Alkyl), C _1-6Alkoxyl group and halo (C _1-6Alkoxyl group) substituting group replaces.

In some embodiments, R ¹, R ², R ³And R ⁴Can randomly be independently selected from hydroxyl, halogen, C by one to three _1-6Alkyl, halo (C _1-6Alkyl), C _1-6Alkoxyl group and halo (C _1-6Alkoxyl group) substituting group replaces.

In some embodiments, R ¹Be independently selected from halogen, C _1-6Alkoxyl group, halo (C _1-6Alkyl), hydroxyl, halo (C _1-6Alkoxyl group), halo (C _1-6Alkyl) alkylsulfonyl, heteroaryl, halo (C _1-6Alkyl) sulfenyl, Heterocyclylalkyl and (C _1-6Alkyl) Heterocyclylalkyl.In some such embodiments, a is 1 and R ¹Be independently selected from 2-chlorine, 2-ethyl, 2-trifluoromethyl, 3-trifluoromethyl, 4-trifluoromethyl, 3-tert-butyl, 4-tert-butyl, 3-ethyl, 4-ethyl, 4-chlorine, 4-bromine, 4-trifluoromethoxy, 4-trifluoromethylthio, 4-trifyl and 4-(4-N-METHYL PIPERAZINE base).In other embodiments, a is 2 and each R ¹Be independently selected from 2-fluorine, 2-chlorine, 2-hydroxyl, 2-methoxyl group, 3-methoxyl group, 5-methoxyl group, 4-chlorine, 4-fluorine, 3-trifluoromethyl, 4-trifluoromethyl, 5-trifluoromethyl, 5-pyridyl, 5-pyridyl-3-base, 5-pyridyl-4-base, 3-THF-3-base, 3-sec.-propyl, 5-sec.-propyl and 5-tert-butyl.

In some embodiments, R ⁴Be selected from C _1-6Alkyl, hydroxyl (C _1-6Alkyl), halo (C _1-6Alkyl), halo (C _1-6Alkyl) sulfenyl, (C _1-6Alkoxyl group) carbonyl, (C _1-6Alkyl) Heterocyclylalkyl, cyanic acid, phenyl, halo (C _1-6Alkyl) phenyl, (C _1-6Alkyl) Heterocyclylalkyl carbonyl and hydroxyl (C _1-6Alkyl amino-carbonyl).In some such embodiments, c is 1 and R ⁴Be selected from trifluoromethyl, cyanic acid, phenyl, trifluoromethylthio, methoxycarbonyl, 4-ethyl piperazidine base, 4-ethyl piperazidine base-1-carbonyl or 2-hydroxyethyl aminocarboxyl.In other embodiments, c is 2 and each R ⁴Be independently selected from methyl, 3-trifluoromethyl, 4-trifluoromethyl, trifluoromethyl, ethoxy carbonyl, hydroxymethyl and phenyl.

In some other embodiment, a kind of compound or its pharmaceutically useful salt are provided, wherein this compound has following formula:

Or have tautomer or the pharmaceutically useful salt of this tautomer of this compound of following formula:

The method of the illness relevant with Raf that the human or animal that the present invention provides treatment to need such treatment on the other hand is individual, it comprises to said individuality uses formula (I), (II), (III) or the compound (IV) that can effectively reduce or prevent individual tumor growth in vivo quantity.

The method of the illness relevant with Raf that the human or animal that the present invention also provides treatment to need such treatment on the other hand is individual, it comprises to said individuality uses formula (I), (II), (III) or compound (IV) and at least a other promoting agent that is used to treat cancer that can effectively reduce or prevent individual tumor growth in vivo quantity.

The method of the illness relevant with Raf that the human or animal that the present invention also provides treatment to need such treatment on the other hand is individual, it comprises to said individuality uses formula (I), (II), (III) or compound (IV) and at least a other promoting agent that is used to treat cancer that can effectively reduce or prevent individual tumor growth in vivo quantity.Think that the many suitable carcinostatic agent that is used as combination therapy all can be used for method of the present invention.In fact, the present invention considers that the many carcinostatic agents that give for example are without limitation: the promoting agent of cell death inducing; Polynucleotide class (for example, ribozyme class); Polypeptide class (for example, enzyme); Medicine; Biology is intended like thing; Alkaloids; The alkylating agent class; Antitumor antibiotics; Metabolic antagonist; Hormones; Platinic compound; The monoclonal antibody of closing with anticarcinogen, toxin and/or radionuclide yoke; Biological response properties-correcting agent (for example interferons [for example IFN-a etc.] and interleukin class [for example IL-2 etc.] etc.); The adoptive immunotherapy agent; The growth factor of hematopoiesis; The promoting agent of inducing tumor cell differentiation (for example alltrans-vitamin A acid etc.); Gene therapy reagent; Antisense therapy reagent and Nucleotide; Tumor vaccine; The suppressor factor that blood vessel takes place etc.Be suitable for the chemotherapy compound of disclosed formula (I), (II), (III) or compound co-administered (IV) and many other instances of anticancer therapy be well known by persons skilled in the art.

In some preferred embodiments, comprise with the carcinostatic agent of The compounds of this invention coupling and inducing or the promoting agent of irritation cell apoptosis.The promoting agent of cell death inducing comprises radiation without limitation; SU11752 (for example, EGF-R ELISA [EGFR] SU11752, angiogenesis factor acceptor [VGFR] SU11752, fibroblast growth factor acceptor [FGFR] SU11752, platelet derived growth factor receptor [PGFR] I SU11752 and Bcr-Abl SU11752 such as STI-571, Gleevec and Glivec]); Antisense molecule; Antibody [for example, Trastuzumab and Rituximab (Rituxan)]; Anti-estrogen [for example, raloxifene and tamoxifen]; Anti--male sex hormone [for example, flutamide, bicalutamide, finasteride, aminoglutethimide, KETOKONAZOL and reflunomide]; Cyclooxygenase 2 (COX-2) suppressor factor [for example, plug is examined west, meloxicam, NS-398 and NSAIDs (NSAID)]; With cancer chemotherapy medicine [for example, Rinotecan (Camptosar), CPT-11, fludarabine (Fuda China (Fludara)), dicarbazine (DTIC), DEXAMETHASONE BP98, mitoxantrone, Mylotarg, VP-16, cis-platinum, 5-FU, Zorubicin, TX or taxol]; The cell signal molecule; Ceramide and cytokine; With star spore rhzomorph etc.

The present invention provides on the other hand and has comprised at least a formula (I), (II), (III) or compound (IV) or its pharmaceutically useful salt and the pharmaceutical composition that is suitable for delivering medicine to the individual pharmaceutically acceptable carrier of human or animal, and this pharmaceutical composition only comprises said compound or also comprises other carcinostatic agent.

The present invention provides on the other hand and has prepared formula described here (I), (II), (III) or (IV) method of compound.

The present invention also provides the compound as the Raf SU11752 on the other hand.Because this enzyme is p21 ^RasThe downstream effect device, therefore suppressor factor of the present invention can be used for wherein need suppressing among the people or animal doctor's pharmaceutical composition of raf kinase pathways, for example, can be used for treating kinase mediated tumour and/or growth of cancer cells by Raf.Because the depending on said Ras protein signal transductory cascade and therefore receive easily through suppressing the Raf kinase activity and block the influence of the treatment that said cascade carries out of these cancers; So compound of the present invention particularly can be used for treating the human or animal; For example, murine cancer.Therefore; Compound of the present invention can be used for treating the entity cancer, as, for example; Cancer (the for example cancer of lung, pancreas, Tiroidina, bladder or colon), marrow illness are (for example; Myelocytic leukemia, multiple myeloma and erythroleukemia), adenoma (for example, fine hair shape adenoma of colon) or sarcoma (for example, osteosarcoma).

Here used " Raf suppressor factor " refers to and with regard to the Raf kinase activity, when measuring with the general Raf/Mek filtration test of describing of hereinafter, shows the IC that is not higher than about 100 μ M and more typically is not higher than about 50 μ M ₅₀Compound.The compounds of this invention will show inhibiting preferred Raf kinases hypotype to it and comprise A-Raf, B-Raf and C-Raf (Raf-1)." IC ₅₀" be that enzyme (for example, Raf kinases) activity is reduced to the half the inhibitor concentration of highest level.Have been found that representative compound of the present invention shows the inhibition activity to Raf.When measuring in the described here Raf kinase assay; Compound of the present invention preferably shows the activity that is not higher than about 10 μ M to Raf; More preferably show and be not higher than about 5 μ M; Even more preferably show and be not higher than about 1 μ M, and most preferably show the activity that is not higher than about 200nM.

Here used phrase " MAPK signal transduction pathway " is the abbreviation of representing mitogen activated protein kinase signal transduction pathway in the module that is formed by the Ras-Raf-MEK1-ERK signaling molecule.

" alkyl " refers to and do not comprise heteroatomic saturated hydrocarbyl and comprise straight chained alkyl such as methyl, ethyl, propyl group, butyl, amyl group, hexyl, heptyl, octyl group, nonyl, decyl, undecyl, dodecyl etc.Alkyl also comprises the branched chain isomer of straight chained alkyl, for example comprises following group without limitation :-CH (CH ₃) ₂,-CH (CH ₃) (CH ₂CH ₃) ,-CH (CH ₂CH ₃) ₂,-C (CH ₃) ₃,-C (CH ₂CH ₃) ₃,-CH ₂CH (CH ₃) ₂,-CH ₂CH (CH ₃) (CH ₂CH ₃) ,-CH ₂CH (CH ₂CH ₃) ₂,-CH ₂C (CH ₃) ₃,-CH ₂C (CH ₂CH ₃) ₃,-CH (CH ₃)-CH (CH ₃) (CH ₂CH ₃) ,-CH ₂CH ₂CH (CH ₃) ₂,-CH ₂CH ₂CH (CH ₃) (CH ₂CH ₃) ,-CH ₂CH ₂CH (CH ₂CH ₃) ₂,-CH ₂CH ₂C (CH ₃) ₃,-CH ₂CH ₂C (CH ₂CH ₃) ₃,-CH (CH ₃) CH ₂CH (CH ₃) ₂,-CH (CH ₃) CH (CH ₃) CH (CH ₃) ₂,-CH (CH ₂CH ₃) CH (CH ₃) CH (CH ₃) (CH ₂CH ₃) etc.Therefore, alkyl comprises primary alkyl, secondary alkyl and tertiary alkyl.Phrase " C _1-12Alkyl " refer to alkyl with 1 to 12 carbon atom.Phrase " C _1-6Alkyl " refer to alkyl with 1 to 6 carbon atom.

" alkenyl " refers to have 2 to 6 carbon atoms and preferably has 2 to 4 carbon atoms and the straight or branched alkyl with at least 1 and the unsaturated position of preferred 1 to 2 vinyl (＞C=C＜).Such group for example has vinyl, allyl group and fourth-3-alkene-1-base.The cis and trans-isomer(ide) or the mixture that comprise these isomer in this term.

" alkoxyl group " refers to wherein, and R is the RO-of alkyl.Here used phrase " C _1-6Alkoxyl group " refer to wherein that R is C _1-6The RO-of alkyl.C _1-6The representative instance of alkoxyl group comprises methoxyl group, oxyethyl group, uncle-butoxy etc.

" (C _1-6Alkoxyl group) carbonyl " refer to wherein that R is C _1-6Ester-the C of alkyl (=O)-OR.

" amidino groups " refers to group-C (=NH) NH ₂" amidine " refers to the compound that comprises such group.

" aminocarboxyl " refers to group-C (O)-NH here ₂

" C _1-6Alkyl amino-carbonyl " refer to group-C (O)-NRR ', wherein R is C _1-6Alkyl and R ' are selected from hydrogen and C _1-6Alkyl.

" carbonyl " refer to divalent group-C (O)-.

" carboxyl " refer to-C (=O)-OH.

" cyanic acid ", " formonitrile HCN " or " nitrile " refer to-CN.

" nitrile (C _1-6Alkyl) " refer to by-substituted the C of CN _1-6Alkyl.

" naphthenic base " refers to single-or multi-ring alkyl substituting group.Typical naphthenic base has 3 to 8 ring carbon atoms.Representational naphthenic base comprises cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl and ring octyl group.

" halogen " or " halo " refers to chlorine, bromine, fluorine and iodine group.

" halo (C _1-6Alkyl) " refer to by one or more preferred 1 to 5 substituted C of halogen atom _1-6Alkyl.Preferred halo (C _1-6Alkyl) is trifluoromethyl.

" halo (C _1-6Alkyl) phenyl " refer to by halo (C _1-6Alkyl) substituted phenyl.

" halo (C _1-6Alkoxyl group) " refer to by one or more preferred 1 to 5 substituted alkoxyl group of halogen atom.Preferred halo (C _1-6Alkoxyl group) is trifluoromethoxy.

" halo (C _1-6Alkyl) alkylsulfonyl " and " halo (C _1-6Alkyl) sulfenyl " refer to by halo (C _1-6Alkyl) substituted alkylsulfonyl and sulfenyl, the wherein definition of alkylsulfonyl and sulfenyl such as described here.

" heteroaryl " refers to that in aromatic ring, to have 1 to 4 heteroatoms and remaining annular atoms as annular atoms be the aromatic group of carbon atom.Suitable heteroatoms used in The compounds of this invention is nitrogen, oxygen and sulphur, and wherein nitrogen and sulphur atom can be randomly oxidized.Instance with heteroaryl of 5 to 14 annular atomses for example comprises, benzimidazolyl-, benzothiazolyl, benzo

azoles base, diaza

base, furyl, pyrazinyl, pyrazolyl, pyridyl, pyridazinyl, pyrimidyl, pyrryl,

azoles base, different

azoles base, imidazolyl, indyl, indazolyl, quinolyl, isoquinolyl, quinazolyl, quinoxalinyl, thiazolyl, thienyl and triazolyl.

" Heterocyclylalkyl " here refer in ring structure, have 1 to 5 and more typical be 1 to 2 heteroatomic naphthenic substituent.Used suitable heteroatoms is nitrogen, oxygen and sulphur in The compounds of this invention, and wherein nitrogen and sulphur atom can be randomly oxidized.Typical Heterocyclylalkyl partly comprises for example morpholino, piperazinyl, piperidyl etc.

" (C _1-6Alkyl) Heterocyclylalkyl " refer to by C _1-6The substituted Heterocyclylalkyl of alkyl.

" Heterocyclylalkyl (C _1-6Alkyl) " refer to by the substituted C of Heterocyclylalkyl _1-6Alkyl.

" Heterocyclylalkyl carbonyl " refers to wherein R here ¹⁰Be group-C (the O)-R of Heterocyclylalkyl ¹⁰

" (C _1-6Alkyl) Heterocyclylalkyl carbonyl " refer to wherein R ¹¹Be (C _1-6Alkyl) group-C of Heterocyclylalkyl (O)-R ¹¹

" hydroxyl " refers to-OH.

" hydroxyl (C _1-6Alkyl) " refer to by the substituted C of hydroxyl _1-6Alkyl.

" hydroxyl (C _1-6Alkyl amino-carbonyl) " refer to by the substituted C of hydroxyl _1-6Alkyl amino-carbonyl.

" imidoether " refers to group-C (=NH) O-or comprise the compound of such group.Imidoether for example comprises imidic acid methyl esters-C (=NH) OCH ₃

" nitro " refers to-NO ₂

" alkylsulfonyl " refers to group-SO here ₂-.

" sulfenyl " refers to group-S-here." alkyl sulphonyl " refers to wherein R ¹²The structural formula that is alkyl is-SO ₂R ¹²Substituted alkylsulfonyl." alkyl sulfenyl " refers to wherein R ¹²The structural formula that is alkyl is-SR ¹²Substituted sulfenyl.Alkyl sulphonyl and alkylthio used in The compounds of this invention comprise (C _1-6Alkyl) alkylsulfonyl and (C _1-6Alkyl) sulfenyl.Therefore, typical group for example comprises methylsulfonyl and methylthio group (that is R wherein, ¹²Be methyl), ethylsulfonyl and ethylmercapto group (that is R wherein, ¹²Be ethyl), third alkylsulfonyl and rosickyite base (that is R wherein, ¹²Be propyl group) etc.

" hydroxy-protective group " refers to the blocking group of OH group.Here this used term also refers to the protection to the OH base of sour COOH.Suitable hydroxy-protective group and be used for specific functional group protected with de-protected suitable condition and be well known in the prior art.For example, at T.W.Greene and P.G.M.Wuts, the blocking group in the organic synthesis (Protecting Groupsin Organic Synthesis), the 3rd edition, Wiley, New York is described a lot of such blocking groups in 1999.Such hydroxy-protective group comprises C _1-6Alkyl oxide, benzyl oxide, right-the methoxyl group benzyl oxide, silyl ether etc.

Term " polymorphic form " refers to the different crystalline form of a kind of compound.Polymorphic form possibly differ from one another aspect many physical propertiess, and is for example different aspect its x-ray diffraction pattern, infrared absorpting light spectra, fusing point, stability or solubleness.

" metabolite " refers to any verivate that in individual's body, produces after with the parent compound administration.Can produce verivate through the for example oxidation of various biochemical transformations, reduction, hydrolysis or the conjugation cause parent compound that in individual's body, carries out and comprise for example oxide compound and demethylation verivate.The metabolite that is equivalent to this analog derivative can also or prepare through compound method with various in vitro methods.In some embodiments, the metabolite of formula (I)-(IV) compound is an oxide compound.Aspect some, this oxide compound is the N-oxide compound, and it is through with oxygenant the compound of formula (I)-(IV) being handled synthetic formation.Aspect some, said oxygenant is N-methylmorpholine N-oxide compound or hydroperoxide such as hydrogen peroxide.In some embodiments, the compound and the glucuronic acid yoke of formula (I)-(IV) closed, thereby form a kind of metabolite.On the other hand, metabolite, tautomer or its steric isomer with following structure is provided:

" randomly substituted " or " substituted " refer to unit price or divalent group and replace one or more Wasserstoffatomss.

When substituted substituting group comprises a kind of straight chain group, can in chain, replace (for example 2-hydroxypropyl, the amino butyl of 2-etc.) or can replace (for example 2-hydroxyethyl, 3-cyanic acid propyl group etc.) at the end of this chain.Substituted substituting group can be covalently bound carbon or heteroatomic straight chain, side chain or circular permutation.

Should be understood that top definition does not comprise impossible replacement mode (for example, by 5 substituted methyl of fluorine or by the substituted halogen atom of another halogen atom).Impossible replacement mode is well-known to those skilled in the art.

Therefore it is obvious to the skilled person that compound of the present invention (comprising formula (I), (II), (III) or compound (IV) or its steric isomer and polymorphic form and the pharmaceutically useful salt of any this compounds, ester, metabolite and prodrug) so can carry out tautomerization and can exist with the prototropy of an atom in the molecule wherein many tautomeric forms to the rearrangement of the chemical bond between another atom and this molecular atoms.See, for example, March, Advanced Organic Chemistry: reaction, mechanism and structure (Advanced Organic Chemistry:Reactions, Mechanisms and Structures), the 4th edition, John Wiley & Sons, 69-74 page or leaf (1992).Terminology used here " tautomer " refers to the compound that moves generation through proton, as long as and should be understood that and can exist, then comprise all tautomeric forms in the present invention.For example, the tautomer of following formula (Ia) compound (it is formula (I) compound of c=1) is the compound of formula (Ib).Similarly, the tautomer of formula (IVc) compound is formula (IVd) or compound (IVe).

Compound of the present invention (comprise formula (I), (II), (III) or compound (IV) or its tautomer and polymorphic form with and any pharmaceutically useful salt, ester, metabolite and prodrug) can comprise by asymmetric substituted carbon atom.Such can be made compound of the present invention with enantiomer, diastereomer and other stereoisomeric forms in any ratio that can define according to the absolute stereo chemistry by asymmetric substituted carbon atom, exists like (R)-or (S)-form.Therefore, comprise single steric isomer, its mixture, the racemic mixture (or " racemic modification ") of all such possible isomer, its optical purity form of The compounds of this invention, the mixture and the one diastereomer of diastereomer in the present invention.Terminology used here " S " and " R " configuration are by IUPAC 1974RECOMMENDATIONS FOR SECTION E, FUNDAMENTALSTEREOCHEMISTRY, Pure Appl.Chem.45:13-30 (1976) definition.α and β use a technical term for the ring position of ring compound.α-the side of reference plane is sides that preferred substituted is positioned at lower label location above that.Being positioned at these substituting groups on the opposite side of these reference plane is designated as with β and describes.Should be noted that this usage is different with usage in the ring-type stereoparent (stereoparente), wherein " α " refers to " being positioned under the said plane " and representes absolute configuration.Terminology used here α and beta comfiguration are that the 203rd section by CHEMICAL ABSTRACTSINDEX GUIDE-APPENDIX IV (1987) defines.

Also it is obvious that to those skilled in the art, and compound of the present invention (comprise formula (I), (II), (III) or compound (IV) or its steric isomer and tautomer with and any pharmaceutically useful salt, ester, metabolite and prodrug) can be with various crystalline form (or " polymorphic form ") forms existence with different physical properties.Should be understood that; As long as it can exist, then be included as all polymorphic forms of the The compounds of this invention (comprise its metabolite, prodrug, steric isomer and tautomer with and any pharmaceutically useful salt) of its unpack format or its form of mixtures among the present invention.

Terminology used here " pharmaceutically useful salt " refers to the nontoxic acid or the alkaline earth salt of formula (I), (II), (III) or compound (IV), tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug.These salt can prepare on the spot in final separation of formula (I), (II), (III) or compound (IV) and purge process or can prepare through in addition alkali or acid functional group being reacted with organic or inorganic acid that suits or alkali.Representational salt comprises following salt without limitation: acetate; Adipate; Alginate; Citrate trianion; Aspartate; Benzoate; Benzene sulfonate; Hydrosulfate; Butyrates; Camphorate; Camsilate; Digluconate; Cyclopentane propionate; Dodecyl sulfate; Esilate; Gluceptate; Glycerophosphate; Hemisulphate; Enanthate; Hexanoate; Fumarate; Hydrochloride; Hydrobromide; Hydriodide; The 2-isethionate; Lactic acid salt; PHENRAMINE MALEATE; Mesylate; Nicotinate; The 2-naphthalenesulfonate; Oxalate; Pamoate; YM 115H salt; Persulphate; 3-phenylpropionic acid salt; Picrate; Pivalate; Propionic salt; SUMATRIPTAN SUCCINATE; Vitriol; Tartrate; Thiocyanate-; Right-tosylate and undecane hydrochlorate.Can also use such as low alkyl group halogen such as methyl, ethyl, propyl group and Butyryl Chloride, bromine and iodine; It is quaternized that the material of dialkylsulfates such as dimethyl-, diethylammonium, dibutyl and diamyl sulfuric ester, long-chain halogenide such as decyl, lauryl, myristyl and stearyl chloride, bromine and iodine, phenylalkyl halogenide such as benzyl and phenethyl bromide etc. and so on will comprise the group of basic nitrogen.Thereby obtain water or the solvable or dispersible products of oil.

The instance that can be used for forming the acid of pharmaceutically useful acid salt comprises mineral acid example hydrochloric acid, sulfuric acid and phosphoric acid and organic acid such as oxalic acid, toxilic acid, methylsulfonic acid, succsinic acid and Hydrocerol A.The alkalescence additive salt can prepare on the spot in the last separation of formula (I) compound and purge process or can be in addition prepare through oxyhydroxide, carbonate or the supercarbonate of carboxylic moiety and alkali that suits such as pharmaceutically acceptable metallic cation or ammoniacal liquor, organic primary, second month in a season or tertiary amine are reacted.Pharmaceutically useful salt comprises the salt of the cationic salt that is the basis with basic metal and earth alkali metal such as sodium, lithium, potassium, calcium, magnesium, aluminium salt etc. and nontoxic ammonium, quaternary ammonium and amine positively charged ion (comprising ammonium, tetramethyl-ammonium, tetraethyl ammonium, methylamine, n n dimetylaniline, Trimethylamine 99, triethylamine, ethamine etc. without limitation) without limitation.Other the typical organic amine that is used to form base addition salt comprises diethylamine, quadrol, thanomin, diethylolamine, piperazine etc.

(the US sequence number is 60/832715 to the title that is on July 21st, 2006 application for the provisional application of " preparation of benzoglyoxaline pyridyl ethers "; Attorney PP028237.0001) and in the title of on August 30th, 2006 application provisional application (attorney PP028258.0001), also disclose the salt and the preparation of The compounds of this invention, it here all is incorporated herein by reference separately for " salt of benzoglyoxaline pyridyl ethers with and preparation ".

Terminology used here " pharmaceutically useful ester " refers to the ester that can be hydrolyzed in vivo, thereby and comprises that those can easily decompose the material that obtains parent compound or its salt in human body.Suitable ester group comprises those that for example derive from pharmaceutically useful aliphatic carboxylic acid, particularly paraffinic acid, alkenoic acid, naphthenic acid and alkanedioic acid, and wherein each alkyl or alkenyl part advantageously has and is not higher than 6 carbon atoms.The instance of concrete ester comprises manthanoate, acetic ester, propionic ester, butyric ester, propenoate and ethyl succinate.

Terminology used here " pharmaceutically useful prodrug " refer in rational medicine judgement scope, be suitable for contacting, do not have over-drastic toxicity, pungency, anaphylaxis etc. with zootic tissue with the people, have rational benefit/risk than and as far as it effective The compounds of this invention of required application prodrug and in possible situation the zwitterionic form of The compounds of this invention.The compound of the parent compound of structural formula above for example hydrolysis produces in blood can easily be changed in vivo in term " prodrug " thereby refer to.At T.Higuchi and V.Stella; " as the prodrug (Pro-drugs as Novel Delivery Systems) of novel transfer system "; The 14th volume and the Edward B.Roche of the A.C.S.Symposium Series edit; " bioreversible carrier in the medicinal design (Bioreversible Carriers in Drug Design) "; American PharmaceuticalAssociation and Pergamon Press provides detailed discussion in 1987, and it here all is introduced into as a reference.

It will be apparent for a person skilled in the art that compound of the present invention (comprise formula (I), (II), (III) or compound (IV) or its tautomer, prodrug, steric isomer and polymorphic form with and any pharmaceutically useful salt, ester and prodrug) can produce the active pharmacological activity metabolite that has kept as suppressor factor through metabolism in human or animal body or in the cell.Can confirm the active metabolite of The compounds of this invention with routine techniques well known in the prior art.See Bertolini for example, people such as G., J.Med.Chem.40:2011-2016 (1997); Shan, people such as D., JPharm.Sci.86 (7): 765-767; Bagshawe K., Drug Dev.Res.34:220-230 (1995); Bodor, N., Advances in Drug Res.73:224-331 (1984); Bundgaard, H., the design of prodrug (Design of Prodrugs) (Elsevier Press 1985); And Larsen, I.K., the design of prodrug and application; Medicinal design and exploitation (Design and Application ofProdrugs; Drug Design and Development) (people such as Krogsgaard-Larsen edits Harwood Academic Publishers, 1991).Should be understood that each compound that comprises the active metabolite that is The compounds of this invention in the present invention.

Term " cancer " refers to the Cancerous disease that can obtain useful treatment through suppressing kinases, particularly Raf kinases, for example comprises; Solid carcinoma; Like cancer (for example, the cancer of lung, pancreas, Tiroidina, ovary, bladder, breast, prostate gland or colon), melanoma, marrow illness (for example, myelocytic leukemia, multiple myeloma and erythroleukemia), adenoma (for example; Fine hair shape adenoma of colon) and sarcoma (for example, osteosarcoma).

In representative embodiment of the present invention, compound of the present invention for example comprises,

{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(4-trifluoromethyl)-amine,

(2-fluoro-5-pyridin-3-yl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2-fluoro-5-pyridin-4-yl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(4-tert-butyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(3-trifluoromethyl-phenyl)-amine,

(3-ethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl-oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(4-chloro-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(4-ethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl-oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(4-chloro-3-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(4-fluoro-3-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(4-trifluoromethoxy-phenyl)-amine,

(2-fluoro-5-trifluoromethyl-phenyl)-(1-methyl-5-{2-[5-methyl-4-(3-trifluoromethyl-phenyl)-1H-imidazoles-2-yl]-pyridin-4-yl oxygen base }-the 1H-benzimidazolyl-2 radicals-yl)-amine,

(2-fluoro-5-trifluoromethyl-phenyl)-(1-methyl-5-{2-[5-methyl-4-(4-trifluoromethyl-phenyl)-1H-imidazoles-2-yl]-pyridin-4-yl oxygen base }-the 1H-benzimidazolyl-2 radicals-yl)-amine,

2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-5-Trifluoromethyl-1 H-imidazoles-4-ethyl formate,

(2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-5-Trifluoromethyl-1 H-imidazol-4 yl)-methyl alcohol,

2-{4-[1-methyl-2-(4-trifluoromethyl-phenyl amino)-1H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-3H-imidazoles-4-formonitrile HCN,

(3-tert-butyl-phenyl)-{ 1-methyl-5-[2-(5-phenyl-1H-imidazoles-2-yl)-pyridin-4-yl-oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

{ 1-methyl-5-[2-(5-phenyl-1H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethylthio-phenyl)-amine,

(3-tert-butyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

[4-fluoro-3-(tetrahydrochysene-furans-3-yl)-phenyl]-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(4-bromo-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(4-fluoro-3-sec.-propyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(4-trifluoromethylthio-phenyl)-amine,

(2-fluoro-5-sec.-propyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2-fluoro-5-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(5-tert-butyl-2-fluoro-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2-fluoro-5-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-methyl isophthalic acid H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2-chloro-4-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-3H-imidazoles-4-formonitrile HCN,

(5-tert-butyl-2-chloro-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2-fluoro-5-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(4-phenyl-5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2-chloro-5-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(4-phenyl-5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

{ 1-methyl-5-[2-(4-phenyl-5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(3-trifluoromethyl-phenyl)-amine,

(3-ethyl-phenyl)-{ 1-methyl-5-[2-(4-phenyl-5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(4-tert-butyl-phenyl)-{ 1-methyl-5-[2-(4-phenyl-5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2-chloro-5-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2-fluoro-5-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-methyl-4-phenyl-1H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2-chloro-5-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-methyl-4-phenyl-1H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(4-tert-butyl-phenyl)-{ 1-methyl-5-[2-(5-methyl-4-phenyl-1H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

{ 1-methyl-5-[2-(5-methyl-4-phenyl-1H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(3-trifluoromethyl-phenyl)-amine,

(5-tert-butyl-2-fluoro-phenyl)-{ 1-methyl-5-[2-(5-methyl-4-phenyl-1H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

[4-(4-methyl-piperazine-1-yl)-phenyl]-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-3H-imidazoles-4-methyl-formiate,

2-{4-[2-(2-chloro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-5-Trifluoromethyl-1 H-imidazoles-4-ethyl formate,

(2-fluoro-4-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2-chloro-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(2,5-dimethoxy-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(3,5-dimethoxy-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(2-trifluoromethyl-phenyl)-amine,

(2-ethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl-oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

(4-ethyl-piperazine-1-yl)-(2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-the 3H-imidazol-4 yl)-ketone,

2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-3H-imidazoles-4-formic acid (2-hydroxyl-ethyl)-acid amides,

{ 1-ethyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(2-fluoro-5-trifluoromethyl-phenyl)-amine,

(2-fluoro-5-trifluoromethyl-phenyl)-{ 6-methoxyl group-1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine,

{ 6-methoxyl group-1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine,

(4-ethyl-piperazine-1-yl)-(2-{4-[1-methyl-2-(4-trifluoromethyl-phenyl amino)-1H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-the 3H-imidazol-4 yl)-ketone,

{ 1-ethyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(4-trifluoromethyl-phenyl)-amine,

2-{4-[1-methyl-2-(4-trifluoromethyl-phenyl amino)-1H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-3H-imidazoles-4-formic acid (2-hydroxyl-ethyl)-acid amides,

2-{1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-base is amino }-5-trifluoromethyl-phenol and

3-{1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-Ji is amino }-6-trifluoromethyl-phenol;

Or the pharmaceutically useful salt of its tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug or this compound, tautomer, steric isomer, polymorphic form, ester, metabolite or prodrug.

On the other hand, the invention still further relates to the method and the synthetic intermediate that is used for these class methods of preparation formula (I), (II), (III) or compound (IV).

That kind as described in detail later the invention still further relates to method for preparing The compounds of this invention and the synthetic intermediate that is used for these class methods.

Compound method

Schema 1 has been explained the structure of the center diaryl ether part of The compounds of this invention.Compound 1.1 and compound 1.2 are reacted (L wherein ¹Or L ²In one be halogen and L ¹Or L ²In another be OH), thereby form ether 1.3.This coupling can in organic solvent such as acetonitrile or DMSO 99.8MIN., exist carry out under the situation of alkali and can raise or reflux temperature under carry out.Suitable alkali comprises K ₂CO ₃, CaCO ₃, KOH, NaOH or KFAl ₂O ₃(Journal of OrganicChemistry, the 63rd volume, the 18th phase, 1998, the 6338-6343 pages or leaves).Group Q in the compound 1.1 can be NH ₂Or amino precursor such as NO ₂Or protected amino, this amino precursor thereafter can be respectively through reducing or go protection to be converted to amine.Z group in the compound 1.2 can be by one or two R ⁴The substituted imidazolyl of group perhaps can be used for forming the functional group of such imidazolyl.Suitable functional group comprises that aldehyde or any aldehyde precursor are as being converted to the ester or the nitrile of aldehyde thereafter.Can this ester and nitrile group be reduced into aldehyde with reductive agent such as diisobutylaluminium hydride.Z can also be-CH ₂OR ⁵, R wherein ⁵It is hydroxy-protective group.In step after a while, can pass through R ⁵Go to protect and the alcohol of gained is oxidized to aldehyde and expose this aldehyde.As shown in the schema 3, this aldehyde is changed into substituted imidazolyl.Be used to form shown in other method such as schema 6 of substituted imidazolyl.

Schema 1:

Schema 2 has shown the synthetic instance of some diaryl ether.Should be understood that in order to describe, the replacement mode below schema 2 has used: Q is NO ₂, L ¹Be OH, L ²Be Cl, Z is the tert-butyl ester.R wherein ²Be that methyl and b are shown in the synthetic instance such as embodiment 1 of 0 aldehyde 2.7.Can amine 2.1 be changed into alkylamine 2.2 through many known methods.On the one hand, amine 2.1 usefulness diacetyl oxides and formic acid are handled, thereby formed corresponding methane amide, it can be reduced into alkylamine 2.2.Suitable reductive agent comprises and has BF ₃(OCH ₂CH ₃) ₂NaBH under the situation ₄Perhaps, alkylamine 2.2 can be through reacting amine 2.1 and trifluoroacetic anhydride, and corresponding amide is removed the trifluoroacetamide blocking group and synthesized with alkylating agent such as alkylogen alkylation and through handling with alkali such as NaOH.

Muriate 2.5 can form chloride of acid 2.4 through with excessive THIONYL CHLORIDE 97 VPP 2.3 being handled, and then it is contacted with pyridine with two carbonic acid di-tert-butyls, prepares thereby obtain muriate 2.5.The alcohol and the muriate 2.5 of alkylamine 2.2 are carried out coupling under alkaline condition; Thereby obtain ether 2.6; Can it directly be changed into aldehyde 2.7 or can through ester 2.6 is reduced into alcohol, then it is oxidized to aldehyde and transforms in two steps through reducing with diisobutylaluminium hydride.

Schema 2:

Schema 3 has been explained the formation of imidazole ring.With aldehyde 2.7 and X wherein ^bBe=O or=NHOH and R ^4pAnd R ^4qBe H or R independently ⁴(R wherein ⁴Definition as previously mentioned), prerequisite is R ^4pAnd R ^4qIn at least one is R ⁴Compound 3.1 react.Can and there be NH in this reaction in polar solvent such as ETHYLE ACETATE/alcohol mixture ₄Carry out under the situation of OH, thereby obtain compound 3.2.Can pass through with reductive agent such as V-Brite B (Na ₂S ₂O ₄) nitroreduction handled compound 3.2 becomes amine 3.3.

Schema 3:

Schema 4 has been explained the formation of benzoglyoxaline ring.Diamines 3.3 and isothiocyanic acid ester 4.1 are reacted, thereby obtain thiocarbamide 4.2.Handle 4.2 with sweetening agent, thus the compound of the formula of obtaining (I).Term " sweetening agent " refers to and is suitable for accomplishing ring closed material such as FeCl ₃, 2-chloro-1-picoline iodide (Mukaiyama reagent), 2-chloro-1,3-methylimidazole muriate, POCl ₃Or alkylogen such as methyl iodide.Can also use the Mukaiyama reagent (Journal ofOrganic Chemistry, the 70th volume, the 7th phase, 2005, the 2835-2838 pages or leaves) of modification.

Schema 4:

Perhaps, can synthesize compound of the present invention through the order that changes coupled reaction.Schema 5 has been explained as 5.1 and 5.2 the coupling that is used to form ehter bond of the penultimate stride of the five rings nuclear that is used to form complete coupling and has been used to form 5.3 and 3.1 coupling of imidazole ring.For midbody 5.1 and 5.2, L ³Or L ⁴In one be halogen and L ³Or L ⁴In another be OH.These midbodys can be through using suitable parent material and/or blocking group to prepare in suitable reaction sequence as shown in the schema of front.Such factor is those skilled in the art's a technical ability.For example, aldehyde 5.3 can be through going back original preparation with diisobutylaluminium hydride to corresponding nitrile (it is synthetic shown in embodiment 60).According to top schema 3 aldehyde 5.3 and ketone 3.1 are reacted, obtain the compound of formula (I).

Schema 5:

The present invention have the compound of triazole end group can be as shown in the schema 6 be that the compound 6.1 of nitrile reacts with hydrazides 6.2 and prepares through Z wherein.Described in the synthetic instance such as embodiment 60 of compound 6.3.

Schema 6:

Should recognize that imidazoles midbody used in this coupled reaction can prepare with some other route of synthesis.Described in a kind of these class methods such as the schema 7.With Z wherein is that the compound 1.3 of CN changes into wherein that Z is the compound of amidino groups.This conversion can be through with 1.3 and alcoholate, react like methylate and carry out, thereby this nitrile is changed into imidoether, thereby next itself and ammonium reagent such as ammonium acetate or ammonium benzoate are reacted the formation amidine.This amidine and X wherein ^aBe compound 7.2 or its tautomer that the reaction of the compound (Va) of leavings group provides alkylation and cyclisation.With compound 7.2 heating, thereby cause eliminating water (dehydration) and form midbody 7.3.Other dehydration conditions comprises with organic acid such as acetate, methylsulfonic acid, camphorsulfonic acid and trifluoroacetic acid and mineral acid example hydrochloric acid and sulfuric acid to be handled 7.2.Usually carry out four reactions with the form of a pot type sequence: form imidoether, form amidine, alkylation/cyclisation and dehydration.

Schema 7:

Compound of the present invention can be used for the growth of anticancer in vivo and in vitro.This compound can use separately or unite use with pharmaceutically useful carrier or vehicle.Suitable pharmaceutically acceptable carrier or vehicle comprise for example machining agent and drug delivery properties-correcting agent and toughener, for example calcium phosphate, Magnesium Stearate, talcum powder, monose, disaccharides, starch, gelatin, Mierocrystalline cellulose, methylcellulose gum, Xylo-Mucine, glucose, hydroxypropyl-beta-cyclodextrin, Vinylpyrrolidone polymer, low melt wax, ion exchange resin etc. with and any two or more combination.At " Remington ' s PharmaceuticalSciences ", Mack Pub.Co. has described other suitable pharmaceutically useful vehicle in New Jersey (1991), and it here is introduced into as a reference.

The significant quantity of The compounds of this invention generally includes other Raf kinase activity test of confirming easily through test described here, known or those of ordinary skills or inhibition or alleviation through the detecting cancer symptom are enough to suppress the active any amount of Raf discernablely.

Thereby the amount that can combine to produce the activeconstituents of single formulation with carrier substance will change according to the main body and the specific administration mode of being treated.But; Should be understood that; For any particular patient; Specific dosage level will depend on many factors, comprise activity, age, body weight, general health, sex, diet, administration time, route of administration, discharge rate, the drug regimen of used particular compound and the severity of the specified disease of treating.Can easily confirm the treatment significant quantity for giving stable condition and it is in common clinicist's technical ability and judgement scope through normal experiment.

For the object of the invention, the treatment effective dose normally delivers medicine to total per daily dose of main body with single dose or fractionated dose, and its total can be for example 0.001 to 1000mg/kg body weight/day and 1.0 to 30mg/kg body weight/day.Dosage unit compositions can comprise its approximate number quantity of forming per daily dose.

Compound of the present invention can be with the form of the dosage unit preparations that comprises conventional nontoxic pharmaceutically acceptable carrier, auxiliary agent and matrix as required by oral administration, parenteral admin, sublingual administration, through atomizing or suck spraying and carry out administration, rectal administration or topical.Topical also can comprise and uses percutaneous dosing as through skin patch or ionophoresis device.The terminology used here parenteral comprises subcutaneous injection, intravenously, intramuscular, breastbone inner injection or input technology.

Can prepare injectable formulation according to prior art, for example injectable aseptic water-based or oil-based suspension with suitable dispersion agent or wetting agent and suspending agent.Injectable sterile preparation can also be to be arranged in the acceptable nontoxic thinner of parenteral or the aseptic injectable solution or the suspension of solvent, for example can be for being positioned at 1, the solution form in the ammediol.Wherein, spendable acceptable matrix and solvent have water, Ringer's solution and isoosmotic sodium chloride solution.In addition, aseptic expressed oil also commonly used is as solvent or suspension medium.For this reason, can use the expressed oil of any gentleness, comprise synthetic list-or two-glycerate.In addition, find in injectable formulation, can also use fatty acid such as oleic acid.

Can be through with medicine and suitable non-stimulated vehicle such as cocoa butter and the polyoxyethylene glycol suppository for preparing the rectal administration that is used for medicine admixed together; Said vehicle is solid at normal temperatures; But under rectal temperature, be liquid, and therefore will in rectum, melt and discharge medicine.

The solid dosage that is used for oral administration can comprise capsule, tablet, pill, pulvis and particle.In such solid dosage, can activeconstituents and at least a inert diluent such as sucrose, lactose or starch is admixed together.As in the routine practice, such formulation also can comprise other material except that inert diluent, for example lubricant such as Magnesium Stearate.In the situation of capsule, tablet and pill, this formulation also can comprise buffer reagent.Can also prepare tablet and pill with casing.

The liquid dosage form that is used for oral administration comprises and comprises the prior art inert diluent commonly used such as pharmaceutical acceptable emulsion, solution, suspension, syrup and the elixir of water.This based composition also can comprise auxiliary agent, like wetting agent, emulsifying agent and suspending agent, Schardinger dextrins and sweeting agent, correctives and perfume compound.

Compound of the present invention can also carry out administration with the form of liposome.That kind as be known in the art, liposome derive from phosphatide or other lipid material usually.Liposome be through be scattered in list in the water-bearing media-or the liquid crystal of multilayer hydration form.Can use any nontoxic physiology that can form liposome can accept and metabolizable lipid.Except that compound of the present invention, the present composition of liposome form also can comprise stablizer, sanitas, vehicle etc.Preferred lipid is phosphatide and phosphatidylcholine (Yelkin TTS), and it can be natural and synthetic.The method that is used to form liposome is known in the prior art.See, for example, Prescott, Ed., Methods in Cell Biology, XIV volume, Academic Press, New York, N.W., the 33rd page and following successive page or leaf (1976).

Though compound of the present invention can be used as unique active medicine by administration, it also can be used to treat other promoting agent coupling of cancer with one or more.Compound of the present invention can also with known therapeutical agent and carcinostatic agent coupling, and the combination of the disclosed compound of the present invention and other anticarcinogen or chemotherapeutics is also within the scope of the invention.Put into practice (CancerPrinciples and Practice of Oncology) in cancer principle and oncology; V.T.Devita and S.Hellman (editor); The 6th edition (February 15 calendar year 2001), during publishing, Lippincott Williams Wilkins can find the instance of such promoting agent.Whether those of ordinary skills can useful according to the combination that the concrete property and the related cancer of medicine are distinguished promoting agent.Such carcinostatic agent comprises following material without limitation: the promoting agent of the suppressor factor of estrogenic agents, androgen receptor modifier, retinoid receptor regulator, cytotoxin/cytostatics, antiproliferative, prenyl-protein transferase inhibitor, HMG-CoA reductase inhibitor and other angiogenesis inhibitor, cell proliferation and survival signal, cell death inducer and interference cell cycle checkpoint.When with the radiotherapy co-administered, compound of the present invention also is useful.

Therefore; In one embodiment of the invention; Compound of the present invention can also with known carcinostatic agent coupling, said known carcinostatic agent comprises for example estrogenic agents, androgen receptor modifier, retinoid receptor regulator, cytotoxic agent, antiproliferative, prenyl-protein transferase inhibitor, HMG-CoA reductase inhibitor, hiv protease suppressor factor, RTI and other angiogenesis inhibitor.

Estrogenic agents refers to the compound that can disturb or suppress oestrogenic hormon and receptors bind, no matter its mechanism how.The instance of estrogenic agents comprises tamoxifen, raloxifene, idoxifene, LY353381, LY117081, toremifene, fulvestrant, 4-without limitation, and [7-(2; 2-dimethyl--1-oxopropoxy-4-methyl-2-[4-[2-(piperidino) oxyethyl group] phenyl]-2H-1-chromene-3-yl]-phenyl-2; 2-dimethyl propylene acid esters, 4; 4 '-dihydroxy benaophenonel-2,4-dinitrophenyl-hydrazone and SH646.

Androgen receptor modifier refers to the compound that can disturb or suppress male sex hormone and receptors bind, no matter its mechanism how.The representative instance of androgen receptor modifier comprises finasteride and other 5 suppressor factor, RU-23908, flutamide, bicalutamide, liarozole and acetic acid Abiraterone.

The retinoid receptor regulator refers to the compound that disturbs or suppress retinoids and receptors bind.The instance of retinoid receptor regulator comprises that bexarotene, vitamin A acid, 13-cis-vitamin A acid, 9-cis-vitamin A acid, alpha-difluoromethyl ornithine, IX23-7553, trans-N-(4 '-hydroxy phenyl) are looked yellow acid amides (retinamide) and the N-4-carboxyl phenyl is looked yellow acid amides.

Cytotoxin/cytostatics refers to mainly through direct interference cell function or inhibition or interference cell mitotic division and causes necrocytosis or suppress the compound of cell proliferation, comprises related kinase whose suppressor factor, metabolic antagonist in the suppressor factor, mitotic division process of alkylating agent, tumour necrosis factor, intercalator, hypoxemia activatory compound, microtubule suppressor factor/microtubule stabilizer, mitotic kinesins; Biological response properties-correcting agent; Hormone/hormone antagonist therapeutical agent, hemopoieticgrowth factor, monoclonal antibody target therapeutic agent, topoisomerase enzyme inhibitor, proteasome inhibitor and ubiquitin ligase suppressor factor.

The instance of cytotoxic agent comprises sertenef, cachectin, ifosfamide, tasonermin, lonidamine, carboplatin, hexamethyl melamine, pool nimustine, mitolactol, ranomustine, fotemustine, S 254, oxaliplatin, TM, heptan platinum, estramustine, improsulfan tosylate, trofosfamide, nimustine, dibrospidium chloride, pumitepa, Lip river platinum, husky platinum, methylmitomycin, cis-platinum, irofulven, right ifosfamide, cis-amination dichloro (2-methyl-pyridine) platinum, benzyl guanine, glufosfamide, GPX100, (trans without limitation; Trans; Trans)-two-mu-(hexane-1; The 6-diamines)-and mu-[diamines-platinum (II)] two [diamines (chlorine) platinum (II)] tetrachloride, two azacyclopropane base spermine, white arsenic, 1-(11-dodecyl amino-10-hydroxyl undecyl)-3,7-dimethyl xanthine, zorubicin, idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin, pinafide, valrubicin, amrubicin, antineoplaston (antineoplaston), 3 '-deaminizating-3 '-morpholino-13-deoxidation-10-hydroxyl carminomycin, At mycin (annamycin), galarubicin, Elinafide, MEN10755 and 4-demethoxylation-3-deaminizating-3-azacyclopropane base-4-methyl sulphonyl-daunorubicin (seeing WO 00/50032).The representative instance of hypoxemia activatory compound is a Win-59075.Proteasome inhibitor comprises lactacystin and Velcade (bortezomib) without limitation.The instance of microtubule suppressor factor/microtubule stabilizer comprises taxol, LY-099094,3 '; 4 '-two dehydrogenations-4 '-deoxidation-8 '-go vincaleucoblastine, Docetaxel, WF 1360, dolastatin, mivobulin isethionate, auristatin, Cemadotin, RPR109881, BMS184476, Vinflunine, cryptophycin, 2; 3,4,5; 6-five fluoro-N-(3-fluoro-4-p-methoxy-phenyl) benzsulfamide, anhydrous vinealeucoblastine(VLB) (anhydrovinblastine), N; N-dimethyl--L-is valyl-and L-is valyl-N-methyl-L-is valyl-and L-prolyl-L-proline(Pro)-uncle-yulocrotine, TDX258, ebormycine (see for example US patent 6,284,781 and 6; 288,237) and BMS188797.The representative instance of topoisomerase enzyme inhibitor has hycamtin, hycaptamine, irinotecan, rubitecan, 6-ethoxy-c acyl group-3 '; 4 '-O-is outer-tolylene-NSC-5159,9-methoxyl group-N, and N-dimethyl--5-nitropyrazole also [3,4; 5-kl] acridine-2-(6H) propylamine, 1-amino-9-ethyl-5-fluoro-2; 3-dihydro-9-hydroxy-4-methyl-1H, 12H-benzo [de] pyrans also [3 ', 4 ': b; 7]-indolizine also [1; 2b] quinoline-10,13 (9H, 15H) diketone, OSI 211,7-[2-(the N-sec.-propyl is amino) ethyl]-(20S) NSC 94600, BNP1350, BNPI1100, BN80915, BN80942, phosphoric acid VP, teniposide, sobuzoxane, 2 '-dimethylamino-2 '-deoxidation-VP, GL ³31, N-[2-(dimethylamino) ethyl]-9-hydroxyl-5,6-dimethyl--6H-pyrido [4,3-b] carbazole-1-methane amide, asulacrine, (5a, 5aB; 8aa, 9b)-9-[2-[N-[2-(dimethylamino) ethyl]-N-methylamino] ethyl]-5-[4-hydroxyl-3,5-Dimethoxyphenyl]-5,5a; 6,8,8a; Also (3 ', 4 ': 6,7) naphtho-(2 for the 9-hexahydro furyl; 3-d)-1,3-dioxole-6-ketone, 2,3-(methylene-dioxy)-5-methyl-7-hydroxyl-8-methoxyl group benzo [c]-coffee pyridine , 6; 9-two [(2-amino-ethyl) amino] benzo [g] isoquinoline 99.9-5; 10-diketone, 5-(3-amino propyl amino)-7; 10-dihydroxyl-2-(2-hydroxyethyl amino methyl)-6H-pyrazolo [4; 5,1 '-de] acridine-6-ketone, N-[1-[2 (diethylamino) ethylamino]-7-methoxyl group-9-oxo-9H-thioxanthene-4-ylmethyl] methane amide, N-(2-(dimethylamino) ethyl) acridine-4-methane amide, 6-[[2-(dimethylamino) ethyl] amino]-3-hydroxyl-7H-indeno [2,1-c] quinoline-7-ketone and dimesna.At the open WO01/30768 of PCT and WO 01/98278, WO 03/050; 064 (on June 19th, 2003), WO03/050; 122 (on June 19th, 2003), WO 03/049; 527 (on June 19th, 2003), WO 03/049; 679 (on June 19th, 2003), WO 03/049; To mitotic kinesins, be described among 678 (on June 19th, 2003) and WO 03/39460 (on May 15th, 2003) and unsettled PCT application US03/06403 (submitting to), US03/15861 (submitting to), US03/15810 (submitting to), US03/18482 (submitting to) and the US03/18694 (in submission on June 12nd, 2003) like the instance of people's mitotic kinesins KSP suppressor factor on June 12nd, 2003 on May 19th, 2003 on May 19th, 2003 on March 4th, 2003.In one embodiment, the suppressor factor of mitotic kinesins comprises the suppressor factor of KSP, the suppressor factor of MKLP1, the suppressor factor of CENP-E, the suppressor factor of MCAK, the suppressor factor of Kifl4, the suppressor factor of Mphosph1 and the suppressor factor of Rab6-KIFL without limitation.

The kinase whose suppressor factor that in the mitotic division process, relates to comprises aurora (aurora) SU11752, Polo-appearance SU11752 (PLK) (the for example suppressor factor of PLK-1), the suppressor factor of bub-1 and the suppressor factor of bub-R1 without limitation.Antiproliferative comprises sense-rna and DNA oligonucleotide such as G3139, ODN698, RVASKRAS, GEM231 and INX3001; And metabolic antagonist such as NSC-239336, carmofur, Tegafur, pentostatin, doxifluridine, Trimetrexate, fludarabine, capecitabine, Ro 09-1390, cytosine arabinoside octadecyl sodium phosphate, fosteabine sodium hydrate, ZD-1694, paltitrexid, emitefur, tiazofurine (tiazofurin), NSC 127716, Nolatrexed, pemetrexed, Nelzarabine (nelzarabine), 2 '-deoxidation-2 '-methylene radical cytidine, 2 '-fluorine methylene radical-2 '-Deoxyribose cytidine, [5-(2 for N-; 3-dihydro-benzofuryl) alkylsulfonyl]-N '-(3; The 4-dichlorophenyl) urea, N6-[4-deoxidation-4-[N2-[2 (E); 4 (E)-14 carbon two enoyl-s] glycyl is amino]-L-glycerine-B-L-seminose-pyrans heptose base] VITAMIN B4, aplidine, ETM775, troxacitabine, 4-[2-amino-4-oxo-4; 6; 7; 8-tetrahydrochysene-3H-Mi Dingbing [5; 4-b] [1; 4] thiazine-6-base-(S)-ethyl]-2,5-thiophene acyl group (thienoyl)-L-L-glutamic acid, aminopterin, 5 FU 5 fluorouracil, alanosine, 11-ethanoyl-8-(carbamoyloxy methyl)-4-formyl radical-6-methoxyl group-14-oxa--1,11-diaza Fourth Ring (7.4.1.0.0)-14 carbon-2; 4,6-triolefin-9-base acetic ester, sphaerophysine, lometrexol, dexrazoxane, methioninase, 2 '-cyanic acid-2 '-'-deoxy-n 4-palmitoyl-1-B-D-arabinofuranosyl adenin glycosyl (arabinofuranosyl) cytosine(Cyt) and 3-EL-970-2-aldehyde thiosemicarbazone.The instance of monoclonal antibody target therapeutic agent comprises having cytotoxic agent or the radioisotopic therapeutical agent that links to each other with cancer cells specificity or target cell monoclonal antibody specific.The example comprises Bexxar.The HMG-CoA reductase inhibitor is the suppressor factor of 3-hydroxy-3-methyl glutaryl base-CoA reductase enzyme.Can use well-known test in the prior art as coming to confirm that easily the HMG-CoA reductase enzyme is had the active compound of inhibition in US patent 4,231,938 and these methods described in the WO 84/02131.The instance of operable HMG-CoA reductase inhibitor comprises that without limitation the Lip river cuts down its spit of fland (MEVACOR

; See US4; 231; 938,4,294,926 and 4; 319,039), suffering is cut down its spit of fland (ZOCOR

; See US4; 444; 784,4,820,850 and 4; 916,239), general its spit of fland (PRAVACHOL

that cuts down; See US4,346,227,4; 537,859,4,410; 629,5,030,447 and 5; 180,589), fluorine cuts down its spit of fland (LESCOL ; See US 5,354,772,4,911; 165,4,929,437,5,189; 164,5,118,853,5,290; 946 and 5,356,896) and its spit of fland of Ah cutting down (LIPITOR

; See US 5,273,995,4,681,893,5,489,691 and 5,342,952).The structural formula of these that can be used for the inventive method and other HMG-CoA reductase inhibitor is at M.Yalpani; " medicine of reducing cholesterol (Cholesterol Lowering Drugs) ", Chemistry Industry, the 87th page and US patent 4 of 85-89 page or leaf (on February 5th, 1996); 782; Be described in 084 and 4,885,314.In one embodiment, the HMG-CoA reductase inhibitor is selected from lovastatin and SV.

Prenyl-protein transferase inhibitor is the compound that can suppress the combination of any prenyl-protein transferase or any prenyl-protein transferase; Said enzyme comprises farnesyl-protein transferase (FPTase), I type geranyl geranyl-protein transferase (GGPTase-I) and II type geranyl geranyl-protein transferase (GGPTase-1I also is called as RabGGPTase).The examples for compounds of inhibition of isoprenyl base-protein transferase comprise (±)-6-[amino (4-chloro-phenyl-) (1-methyl isophthalic acid H-imidazoles-5-yl) methyl]-4-(3-chloro-phenyl-)-1-methyl-2 (1H)-quinolinone, (-)-6-[amino (4-chloro-phenyl-) (1-methyl isophthalic acid H-imidazoles-5-yl) methyl]-4-(3-chloro-phenyl-)-1-methyl-2 (1H)-quinolinone, (+)-6-[amino (4-chloro-phenyl-) (1-methyl isophthalic acid H-imidazoles-5-yl) methyl]-4-(3-chloro-phenyl-)-1-methyl-2 (1H)-quinolinone, 5 (S)-just-butyl-1-(2, the 3-3,5-dimethylphenyl)-4-[1-(4-cyanic acid benzyl)-5-imidazolyl methyl-2-piperazine ketone, (S)-1-(3-chloro-phenyl-)-4-[1-(4-cyanic acid benzyl)-5-imidazolyl methyl]-5-[2-(ethylsulfonyl) methyl)-2-piperazine ketone, 5 (S)-just-butyl-1-(2-aminomethyl phenyl)-4-[1-(4-cyanic acid benzyl)-5-imidazolyl methyl]-2-piperazine ketone, 1-(3-chloro-phenyl-)-4-[1-(4-cyanic acid benzyl)-2-methyl-5-imidazolyl methyl]-2-piperazine ketone, 1-(2, the 2-diphenyl-ethyl)-3-[N-(the basic ethyl of 1-(4-cyanic acid benzyl)-1H-imidazoles-5-) formamyl] piperidines, 4-{-[4-hydroxymethyl-4-(4-chloropyridine-2-ylmethyl)-piperidines-1-ylmethyl]-glyoxal ethyline-1-ylmethyl cyanobenzene, 4-{-5-[4-hydroxymethyl-4-(3-benzyl chloride base)-piperidines-1-ylmethyl]-glyoxal ethyline-1-ylmethyl-cyanobenzene, 4-{3-[4-(2-oxo-2H-pyridine-1-yl) benzyl]-3H-imidazol-4 yl methyl cyanobenzene, 4-{3-[4-(5-chloro-2-oxo-2H-[1; 2 '] two pyridines-5 '-ylmethyl]-3H-imidazol-4 yl-methyl } cyanobenzene, 4-{3-[4-(2-oxo-2H-[1,2 '] two pyridines-5 '-ylmethyl]-3H-imidazoles 4-base-methyl } and cyanobenzene, 4-[3-(2-oxo-1-phenyl-1,2-dihydropyridine-4-methyl)-3H-imidazol-4 yl methyl } cyanobenzene, 18; 19-dihydro-19-oxo-5H, 17H-6,10:12; 16-dimethylene-1H-imidazo [4,3-c] [1,11; 4] dioxa nitrogen heterocyclic 19 alkynes-9-nitrile, (+-)-19,20-dihydro-19-oxo-5H-18,21-ethano--12; 14-vinylidene-6; 10-methylene radical-22H-benzo [d] imidazo [4,3-k]-[1,6; 9; 12] oxa-three azepines-ring octadecyne-9-nitrile, 19,20-dihydro-19-oxo-5H; 17H-18; 21-ethano--6,10:12,16-dimethylene-22H-imidazo [3; 4-h] [1; 8,11,14] oxa-three nitrogen heterocyclics 20 carbynes-9-nitrile and (+-)-19; 20-dihydro-3-methyl isophthalic acid 9-oxo-5H-18; 21-ethano--12,14-vinylidene-6,10-methylene radical-22H-benzo [d] imidazo [4; 3-k] [1; 6,9,12] oxa-three nitrogen heterocyclics octadecyne-9-nitrile.Can find other instance of prenyl-protein transferase inhibitor: WO 96/30343, WO97/18813, WO 97/21701, WO 97/23478, WO 97/38665, WO 98/28980, WO 98/29119, WO 95/32987, US patent 5 in publication below and the patent; 420; 245, the US patent 5; 523; 430, the US patent 5; 532; 359, the US patent 5,510, and 510, US patent 5; 589; 485, the US patent 5,602, and 098, European patent discloses 0 618 221, European patent discloses 0 675 112, European patent discloses 0 604 181, European patent discloses 0 696 593, WO 94/19357, WO 95/08542, WO 95/11917, WO 95/12612, WO 95/12572, WO 95/10514, US patent 5; 661; 152, WO95/10515, WO 95/10516, WO 95/24612, WO 95/34535, WO 95/25086, WO 96/05529, WO 96/06138, WO 96/06193, WO 96/16443, WO 96/21701, WO 96/21456, WO 96/22278, WO 96/24611, WO 96/24612, WO 96/05168, WO 96/05169, WO 96/00736, US patent 5,571,792, WO 96/17861, WO96/33159, WO 96/34850, WO 96/34851, WO 96/30017, WO 96/30018, WO 96/30362, WO 96/30363, WO 96/31111, WO 96/31477, WO 96/31478, WO 96/31501, WO 97/00252, WO 97/03047, WO 97/03050, WO 97/04785, WO 97/02920, WO 97/17070, WO 97/23478, WO 97/26246, WO 97/30053, WO 97/44350, WO 98/02436 and US patent 5; 532,359.For the instance that prenyl-protein transferase inhibitor is had an effect as far as blood vessel, can be referring to European J.ofCancer 55 (9): 1394-1401 (1999).

Angiogenesis inhibitor refers to the compound that can suppress neovascularization, no matter its mechanism how.The instance of angiogenesis inhibitor comprises tyrosine kinase inhibitor without limitation; Suppressor factor, MMP (matrix metalloproteinase) suppressor factor, integrin retarding agent, interferon-' alpha ', il-1 2, piperylene like the suppressor factor of tyrosine kinase receptor Flt-1 (VEGFR1) and Flk-1/KDR (VEGFR2), epidermis deutero-, inoblast deutero-or platelet-derived growth factor gather sulfuric ester, cyclooxygenase-2 inhibitors, comprise that NSAIDs (NSAID) is like Frosst) and Ibuprofen BP/EP and selectivity COX-2 inhibitors such as celecoxib and rofecoxib (PNAS89:7384 (1992); JNCI 69:475 (1982); Arch.Ophthalmol.108:573 (1990); Anat.Rec, (238): 68 (1994); FEBS Letters 372:83 (1995); Clin, Orthop.313:76 (1995); J Mol.Endocrinol.16:107 (1996); Jpn.J.Pharmacol.75:105 (1997); Cancer Res.57:1625 (1997); Cell 93:705 (1998); Intl.J.Mol.Med.2:715 (1998); J Biol.Chem.274:9116 (1999)), steroidal anti-inflammatory medicine (like reflunomide, mineralocorticoid, DEXAMETHASONE BP98, prednisone, prednisolone, methylprednisolone, Betamethasone Valerate), carboxyl amido triazole, examine its spit of fland A-4 of cloth, Squalamine, 6-O-chloracetyl-carbonyl)-his spit of fland of aspergillus fumigatus cedrol, Thalidomide, blood vessel, troponin-1, Angiotensin II antagonist (see people such as Fernandez; J.Lab.Clin.Med.105:141-145 (1985)) and VEGF antibody (see NatureBiotechnology, 17:963-968 (October 1999); People such as Kim, Nature, 362:841-844 (1993); WO 00/44777; With WO 00/61186).The material (seeing the summary of in Clin.Chem.La.Med.38:679-692 (2000), carrying out) of adjustable or anticoagulant and fibrinolytic system takes place and also can comprise with other therapeutical agent of compound associating of the present invention in adjustable or inhibition blood vessel.Can regulation and control or the instance of such material of anticoagulant and fibrinolysis approach comprise heparin (seeing Thromb.Haemost.80:10-23 (1998)), low molecular weight heparin and carboxypeptidase U suppressor factor (suppressor factor that also is called as active enzyme thrombin activatory fibrinolysis inhibitor [TAFIa]) (seeing Thrombosis Res.101:329-354 (2001)) without limitation.In the open WO 03/013,526 of PCT and sequence number are the US patented claim of 60/349,925 (in application on January 18th, 2002), the TAFIa suppressor factor is described.The present invention also comprises The compounds of this invention and is that selective COX-2-2 suppressor factor (is generally defined as COX-2 is had special inhibition (when assessing with cell or microbiological test, with the IC of COX-2 ₅₀The IC of/COX-1 ₅₀Ratio when measuring, at least 100 times of the inhibition height of the inhibition comparison COX-1 of COX-2) material) the associating of NSAID.This compounds is included in US patent 5,474,995 (open December 12 nineteen ninety-five), US patent 5,861,419 (open on January 19th, 1999), US patent 6 without limitation; 001,843 (open), US patent 6,020,343 (open), US patent 5,409 on February 1st, 2000 on December 14th, 1999; 944 (in nineteen ninety-five publication in April 25), US patent 5,436,265 (open), US patent 5,536 July 25 nineteen ninety-five; 752 (open), US patent 5,550,142 (open), US patent 5,604 on August 27th, 1996 on July 16th, 1996; 260 (open), US patent 5,698,584 (publishing), US patent 5,710 December 16 in 1997 on February 18th, 1997; 140 (month January 20 in 1998), WO 94/15932 (open), US patent 5,344,991 (open), US patent 5,134 on June 6th, 1994 July 21 in 1994; 142 (open), US patent 5,380,738 (open), US patent 5,393 January 10 nineteen ninety-five on July 28th, 1992; 790 (open), US patent 5,466,823 (open), US patent 5,633 November 14 nineteen ninety-five February 20 nineteen ninety-five; Disclosed compound in 272 (open on May 27th, 1997) and the US patent 5,932,598 (open on August 3rd, 1999), these documents here all are introduced into as a reference.The typical suppressor factor that is used for the COX-2 of the inventive method comprises 3-phenyl-4-(4-(methylsulfonyl) phenyl)-2-(5H)-furanone; With 5-chloro-3-(4-methylsulfonyl) phenyl-2-(2-methyl-5-pyridyl) pyridine.Find the specific inhibitor that is described to COX-2 in patent that can be below, the application of pending trial and the publication and therefore can be used for compound of the present invention with and compound method (these documents here are introduced into as a reference): WO 94/15932 (open), US patent 5,344,991 (open), US patent 5,134 on June 6th, 1994 on July 21st, 1994; 142 (open), US patent 5,380,738 (open), US patent 5,393 January 10 nineteen ninety-five on July 28th, 1992; 790 (open), US patent 5,466,823 (open), US patent 5,633 November 14 nineteen ninety-five February 20 nineteen ninety-five; 272 (open), US patent 5,932,598 (open), US patent 5,474 on August 3rd, 1999 on May 27th, 1997; 995 (open), US patent 5,861,419 (open), US patent 6,001 on January 19th, 1999 December 12 nineteen ninety-five; 843 (open), US patent 6,020,343 (open), US patent 5,409 on February 1st, 2000 on December 14th, 1999; 944 (open), US patent 5,436,265 (open), US patent 5,536 July 25 nineteen ninety-five April 25 nineteen ninety-five; 752 (open), US patent 5,550,142 (open), US patent 5 on August 27th, 1996 on July 16th, 1996; 604,260 (open), US patent 5,698 on February 18th, 1997; 584 (open) and US patent 5,710,140 (open) on January 20th, 1998 on December 16th, 1997.Other instance of angiogenesis inhibitor comprises his spit of fland of endothelium, ukrain, ranpirnase, IM862,5-methoxyl group 4-[2-methyl-3-(3-methyl-2-butene base) Oxyranyle]-1-oxa-volution [2 without limitation; 5] suffering-6-base (chloracetyl) carbamate, ethanoyl Goe 1734,5-amino-1-[[3; 5-two chloro-4-(4-chlorobenzene formacyl) phenyl] methyl]-1H-1; 2,3-triazole-4-methane amide, CM101, Squalamine, examine its spit of fland of cloth, RPI4610, NX31838, Sulfated phosphoric acid mannopentaose, 7,7-(carbonyl-two [imino--N-methyl-4; 2-pyrrolo-carbonyl imino-[N-methyl-4; 2-pyrroles]-the carbonyl imino-]-two-(1, the 3-napadisilate) and 3-[(2,4-dimethyl pyrrole-5-yl) methylene radical]-2-dihydroindolone (SU5416).

The promoting agent of interference cell cycle checkpoint is the protein kinase that can suppress transducer cell cycle checkpoint signal, thereby can make cancer cells to the responsive compound of DNA infringement agent.Such material comprises the kinase whose suppressor factor of ATR, ATM, Chk1 and Chk2 and cdk and cdc SU11752, and its particular example has 7-hydroxyl star spore rhzomorph, flavones pyrrole alcohol (flavopiridol), CYC202 (Cyclacel) and BMS-387032.

The suppressor factor of cell proliferation and survival signal transduction path is the forms of pharmacologically active agents that suppresses the signal transduction cascade in cell surface receptor and these surface receptor downstream.Such material comprises the suppressor factor (for example ZD1939 (gefitinib) and erlotinib (erlotinib)) of EGFR suppressor factor; The suppressor factor of ERB-2 (for example trastuzumab); The suppressor factor of IGFR; The suppressor factor of cytokine receptor; The suppressor factor of MET; PI3K suppressor factor (for example LY294002); (suppressor factor that comprises Akt without limitation is as at WO 02/083064 for serine/threonine kinase; WO 02/083139; These materials described in WO02/083140 and the WO 02/083138); The kinase whose suppressor factor of Raf (for example BAY-43-9006); The suppressor factor (for example Wyeth CCI-779) of the suppressor factor of MEK (for example CI-1040 and PD-098059) and mTOR.Such material comprises micromolecular inhibitor compound and antibody antagonist.

Cell death inducer comprises the TNF receptor family member acvator of (comprising the TRAIL acceptor).

In some embodiment preferred of the present invention, unite the typical activity agent that is used to treat cancer with The compounds of this invention and comprise for example Rinotecan, hycamtin, gemcitabine, 5 FU 5 fluorouracil, LV, carboplatin, cis-platinum, taxanes, Te Zhata shore (tezacitabine), endoxan, vinca alkaloids, imatinib (Gleevec), anthracycline antibiotics, Rituximab, trastuzumab and other cancer chemotherapeutic agent.

With the top compound of The compounds of this invention coupling will with doctor's handbook (the Physicians ' Desk Reference) (PDR) therapeutic dose shown in the 47th edition (1993) (it here be introduced into as a reference) use, perhaps such treatment application quantity will be that those of ordinary skills are known.

Compound of the present invention can be used with the maximum clinical dosage of recommendation or than low dosage with other carcinostatic agent.Can according to the severity of route of administration, disease and patient respond change active compound in the present composition dosage level to obtain required treatment response.This combination can be carried out administration with compsn independently or with the form of the single formulation that comprises two kinds of promoting agents.When carrying out administration with array configuration, said therapeutical agent can be configured to can be in identical time or different time by the independently compsn of administration, and perhaps said therapeutical agent can be given with the form of single compsn.

Antiestrogen such as tamoxifen have suppressed the breast cancer growth through the inducing cell cycle arrest, and it needs the effect of cell cycle inhibitor p27Kip.Recently; Shown that the activation of Ras-Raf-MAP kinase pathways has changed the phosphorylation state of p27Kip, thereby made it weakened, thereby helped antiestrogen tolerance (people such as Donovan in retardance inhibition activity in the cell cycle; J Biol.Chem.27 (5:40888,2001).As people such as Donovan report, suppress the MAPK signal and changed the phosphorylation state of p27 in the breast cancer cell line that is difficult to treat with hormone and recovered hormone-sensitive thus through treating with mek inhibitor.Therefore; On the one hand; In hormonal dependent cancer such as breast cancer and treatment of prostate cancer, can use formula (I), (II), (III) or (IV) any embodiment of compound or its tautomer, pharmaceutically useful salt or the pharmaceutically useful salt of its tautomer reverse when using conventional carcinostatic agent usually observed hormone tolerance in these cancers.

In the hematology cancer, in chronic granulocytic leukemia (CML), chromosome translocation has caused the composition activation of BCR-AB1 Tyrosylprotein kinase.Because suppressed the Ab1 kinase activity, so receive the ailing patient who torments that Gleevec (a kind of small molecules tyrosine kinase inhibitor) is had response.But, when many patients that suffer from terminal illness begin Gleevec is had response, but afterwards owing to cause the sudden change of the Ab1 kinase domain of tolerance, its disease recurs after a while.Verified in vitro study, BCR-Av1 utilizes the Raf kinase pathways to cause its effect.In addition, the kinases that in identical approach, suppresses more than one provides other provide protection to the tolerance of mutagenesis.Therefore; In another aspect of this invention; In the hematology treatment for cancer; With formula (I), (II), (III) or compound (IV) or its tautomer, its pharmaceutically useful salt or the pharmaceutically useful salt of its tautomer and at least a other promoting agent of any embodiment, as the Gleevec coupling with reverse or prevention to the tolerance of described at least a other promoting agent.

On the other hand; The present invention relates to suppress the method for the individual biology situation by the mediation of the serine/threonine kinase in the MAPK signal transduction pathway of serine/threonine kinase at least a MAPK signal transduction pathway in the individual's body or treatment, it comprises using and comprises active at least a formula (I), (II), (III) or the compound (IV) that can effectively suppress the serine/threonine kinase at least a MAPK signal transduction pathway in patient's body or the therapeutic compsn of tautomer, its pharmaceutically useful salt or the pharmaceutically useful salt of its tautomer.

The therapeutic compsn of this respect of the present invention can be used for treating the patient's (for example, suffer from by MAPK abnormal signal cancers mediated patient) who such suppressor factor is had demand.Comprise for example melanoma, papillary carcinoma, thyroid carcinoma, ovarian cancer, colorectal carcinoma, carcinoma of the pancreas, nonsmall-cell lung cancer (NSCLC), acute lymphoblastic leukemia (ALL) and acute myelocytic leukemia by unusual MAPK signal cancers mediated type.Can suppress unusual MAPK signal through the compound that suppresses wild-type or mutant Ras, Raf, MEK or ERK.

In one embodiment, the invention provides the method for a kind of inhibition Ras (wild-type or mutant Ras).This method comprises formula (I), (II), (III) or compound (IV) or its tautomer, pharmaceutically useful salt or the pharmaceutically useful salt of its tautomer of using any embodiment of significant quantity to its individuality of needs.

In one embodiment, the invention provides the method for a kind of inhibition Raf (wild-type or mutant B-Raf).This method comprises formula (I), (II), (III) or compound (IV) or its tautomer, pharmaceutically useful salt or the pharmaceutically useful salt of its tautomer of using any embodiment of significant quantity to its individuality of needs.

In one embodiment, the invention provides the method for a kind of MEK of inhibition.This method comprises formula (I), (II), (III) or compound (IV) or its tautomer, pharmaceutically useful salt or the pharmaceutically useful salt of its tautomer of using any embodiment of significant quantity to its individuality of needs.

In one embodiment, the invention provides the method for a kind of ERK of inhibition.This method comprises formula (I), (II), (III) or compound (IV) or its tautomer, pharmaceutically useful salt or the pharmaceutically useful salt of its tautomer of using any embodiment of significant quantity to its individuality of needs.

Described in following embodiment 82-86 and 89-90 and as shown in Fig. 6-12B, be used for a kind of exemplary compounds of this method of the present invention---1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine shows the effect of effective inhibition MAPK signal transduction pathway.As shown in Figure 82, in biochemical test, this compound is the suppressor factor of effective B-Raf, c-Raf, mutant B-Raf and mutant Ras, has proved that it is to the active restraining effect (IC of mutant B-Raf ₅₀Be 0.0053 μ M), to the active restraining effect (IC of B-Raf ₅₀Be 0.068 μ M) and to the active restraining effect (IC of c-Raf ₅₀Be 0.004 μ M).The treatment of carrying out with said compound has all caused tumor regression in all three kinds of mutant B-Raf xenograft models (A375M5, MEXF276 and HT29) of being tested; And as summarized in the following table 7 and as described in embodiment 84,85 and 86, in the xenograft models that K-Ras and N-Ras drive, produced the tumor growth inhibition.

As shown in Fig. 7 B, 8B and the 10C, target is regulated the phosphorylation that its mode with dosage and time-dependent manner of analysis revealed of carrying out has suppressed MEK in usefulness { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine is treated afterwards to tumour cell A375M, MEXF276 and HCT-116.In addition, as shown in Fig. 7 D, 8C and the 9C, the downstream affinity tag of Raf is being regulated and control in the treatment of using this compound that tumour cell A375M, MEXF276 and HCT-116 are carried out, comprises BIM, cyclin D1, p27Kip and pAKT.These tests of in preclinical models, carrying out show that { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine all shows the restraining effect of dosage and time-dependent manner to the downstream signaling molecule of Raf in phosphorylation of MEK target and the MAPK approach.

On the other hand; The present invention relates to suppress tyrosine kinase receptor or the treatment of at least a VEGFR-2 of being selected from the individual's body, PDGFR-β, pERK, bFGF, FGFR1, FGFR2, FGFR3, c-Kit and the CSF-1R method by the biology situation of at least a mediation of VEGFR-2, PDGFR-β, pERK, bFGF, FGFR1, FGFR2, FGFR3, c-Kit and CSF-1R, it comprises uses the therapeutic compsn that comprises at least a formula (I), (II), (III) or the compound or pharmaceutically acceptable salt thereof (IV) that can effectively suppress individual intravital tyrosine kinase receptor.

Treatment compound of the present invention can be used for treating the patient's (for example, suffer from by the unusual cancers mediated of tyrosine kinase receptor signal transduction patient) who such suppressor factor is had demand.Comprise for example melanoma, breast cancer, bladder cancer, lung cancer, thyroid carcinoma, prostate cancer, ovarian cancer, mast cell leukemia, germinoma, small cell lung cancer, gastrointestinal stromal tumor, acute myelogenous leukemia (AML), neuroblastoma and carcinoma of the pancreas by the unusual cancers mediated of tyrosine kinase receptor signal transduction.

In one embodiment, the invention provides the method for a kind of VEGFR-2 of inhibition.This method comprises formula (I), (II), (III) or compound (IV) or its pharmaceutically useful salt of using any embodiment of significant quantity to its individuality of needs.This method can be used to treat solid tumor through suppressing blood vessel.

In one embodiment, the invention provides the method for a kind of PDGFR-of inhibition β.This method comprises formula (I), (II), (III) or compound (IV) or its pharmaceutically useful salt of using any embodiment of significant quantity to its individuality of needs.

In one embodiment, the invention provides the method for a kind of c-Kit of inhibition.This method comprises formula (I), (II), (III) or compound (IV) or its pharmaceutically useful salt of using any embodiment of significant quantity to its individuality of needs.

In one embodiment, the invention provides the method for a kind of CSF-1R of inhibition.This method comprises formula (I), (II), (III) or compound (IV) or its pharmaceutically useful salt of using any embodiment of significant quantity to its individuality of needs.

In biochemical test, be used for a kind of exemplary compounds---{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-4-trifluoromethyl-phenyl of this method on the one hand of the present invention)-amine is the suppressor factor of effective tyrosine kinase receptor VEGFR-2, PDGFR-β, pERK, bFGF, FGFR1, FGFR2, FGFR3, c-Kit and CSF-1R.Shown in embodiment 87, this compound shows and suppresses the active (IC of VEGFR-2 ₅₀Be 0.07 μ M), suppress PDGFR-β (IC ₅₀Be 0.0032 μ M), suppress c-Kit (IC ₅₀Be 0.02 μ M) and suppress CSF-1R (IC ₅₀Be 0.20 μ M) effect.In addition, shown in figure 13 and as described in the embodiment 88, the inhibition that this compound has induced blood vessel to take place in matrigel (matrigel) model in vivo.

Through can more easily understanding the present invention with reference to following embodiment, it is in order to describe that these embodiment are provided, and is not to limit the invention with it.

Embodiment below and the application everywhere in, following abbreviation has following implication.If not explanation, then this term has its common art-recognized meanings.

APCI APCI atmospheric pressure chemical ionization mass spectrum

Aq. aqueous

The atm normal atmosphere

Cm centimetre

℃ degree centigrade

The DIPEA diisopropyl ethyl amine

DMC 2-chloro-1,3-methylimidazole quinoline muriate

The DMSO DMSO 99.8MIN.

Eq. equivalent

EtOAc ETHYLE ACETATE

EtOH ethanol

G or gm gram

H/hr/hrs hour

The HPLC HPLC

The IPA Virahol

The L liter

LCAP liquid chromatography area percentage

The LC/MS liquid chromatography-mass spectrography

The M volumetric molar concentration

The MeCN acetonitrile

The mL milliliter

The NaOMe sodium methylate

1-PrOH 1-propyl alcohol

The TEA triethylamine

The TFAA trifluoroacetic anhydride

THF hydrogen furans

The representative side chain that is used for following embodiment compound can prepare according to following method usually:

Embodiment 1

The preparation of { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine

Step 1

Mix mechanical stirrer and add K for the three-necked flask of a 500mL ₂CO ₃(4.15g, 30mmol).With the sealing of this flask, find time and its flame is dry.Make this device be cooled to room temperature and it is used purification for argon.In this reaction flask, add 4-amino-3-nitro phenol 1a (3.08g, 20mmol), (5.2g is 24mmol) with anhydrous DMSO (30mL) for 4-chloropyridine-2-formic acid uncle-butyl ester 1b.With the mixture vigorous stirring of gained and be heated to about 14 hours of 100 ℃ of heating.This reaction is poured in the ice-cold phosphate buffered saline buffer (pH=7) and this reaction flask is fully washed with MTBE (MTBE) and water.The biphasic mixture that is merged is filtered with zeyssatite (pad of＞2cm).This layer is distributed and separates and (3 * 100mL) extract with MTBE with water.With the organic layer that is merged with water washing (5 * 100mL), dry (MgSO ₄) and evaporation.This resistates bullion is adsorbed onto SiO ₂On, and it is carried out purifying (4: 1,2: 1,1: 1 hexane-EtOAc (ETHYLE ACETATE)) with flash column chromatography, obtain the 1c of 4.92g (14.9mmol, yield are 74%) tawny solid form. ¹H?NMR(300MHz，CDCl ₃)δ8.58(d，J＝5.8Hz，1H)，7.90(d，J＝2.8Hz，1H)，7.56(d，J＝2.5Hz，1H)，7.17(dd，J＝2.8，8.8Hz，1H)，6.94(dd，J＝2.8，5.8，Hz，1H)，6.91(d，J＝9.1Hz，1H)，6.15(br?s，2H)，1.62(s，9H)； ¹³C?NMR(75MHz，CDCl ₃)δ165.8，164.0，151.8，151.5，143.4，143.2，131.5，129.8，121.0，118.0，114.2，113.1，83.0，28.4；mp?163-166℃。

Step 2

Under 0 ℃, (5.62g is 17mmol) at CH to N-methyl-p-nitroaniline 1c ₂Cl ₂Add in the solution (85mL) TFAA (2.4mL, 3.6g, 17mmol).Then, remove cooling bath and should react and at room temperature keep 2 hours.With this reaction cooled to 0 ℃ and to wherein add TBACl (tetrabutylammonium chloride, 2.5g, 8.5mmol), Me ₂SO ₄(methyl-sulfate, 3.2mL, 4.3g, 34mmol) and 10%NaOH (34mL).With the mixture of gained vigorous stirring 4 hours at room temperature.Distribute and separate with this reaction dilute with water and to the layer of gained.Water is used CH ₂Cl ₂(3 * 100mL) extract and with the organic layer that is merged with brine wash (2 * 100mL), dry (MgSO ₄) and evaporation.With this resistates bullion be adsorbed onto on the silica gel and with flash column chromatography to its carry out purifying (4: 1,2: 1,1: 1,1: 2 hexane/EtOAc), thereby obtain the 1d of 4.5g (13.0mmol, 76%) Huang-orange solids form. ¹H?NMR(300MHz，CDCl ₃)δ8.54(d，J＝5.5Hz，1H)，8.04(br?d，J＝4.7Hz，1H)，7.93(d，J＝2.8Hz，1H)，7.53(d，J＝2.5Hz，1H)，7.25(app?dd，J＝2.8，9.1Hz，1H)，6.91(m，2H)，3.04(d，J＝4.9Hz，3H)，1.59(s，9H)； ¹³C?NMR(75MHz，CDCl ₃)δ165.9，164.1，151.5，144.7，142.1，130.4，118.8，115.5，114.1，112.9，82.9，30.4，28.5；mp?187-189℃。

Step 3

Use N to one ₂(lithium aluminum hydride, 3.0g is 75mmol) with anhydrous THF (240mL) to add LAH in the flame-dried 500mL three neck round-bottomed flasks that purify.With the suspension of gained be cooled to 0 ℃ and its internal reaction temperature is remained on 5 ℃ slowly add in down tert-butyl ester 1d (20.7g, 60mmol).Then, this reaction mixture was stirred 2 hours down at 0 ℃, then it is at room temperature stirred a night.Add NaBH ₄(2.27g is 60mmol) and with this reaction mixture restir 1 hour at room temperature.After it reacts completely in judgement, drip water (3mL), 15%NaOH (3mL) and water (9mL) in succession this reaction mixture is handled.The mixture of gained is washed with EtOAc and methyl alcohol with diatomite filtration and with remaining solid.Be adsorbed onto SiO with the evaporation of the organic moiety that merged and with the resistates bullion of gained ₂Upward and with flash column chromatography it is carried out purifying (97: 3CH ₂Cl ₂-MeOH), thereby obtain 7.63g (27.7mmol, 46%) red-1e of orange solids form. ¹H?NMR(300MHz，CDCl ₃)δ8.40(d，J＝5.5Hz，1H)，8.05(br?s，1H)，7.96(d，J＝2.75Hz，1H)，7.29(d，J＝2.75Hz，1H)，6.92(d5?J＝9.35Hz，1H)，6.75(m，2H)，4.68(s，2H)，3.07(d，J＝5.23Hz，3H)。

Step 4

In the round-bottomed flask of a 100mL, add benzylalcohol 1e (1.38g, 5.0mmol), MnO ₂(6.52g, 75mmol) and CHCl ₃(20mL).The suspension of gained was stirred 2 days down in room temperature (rt).This reaction mixture is used diatomite filtration, and remaining solid is used CHCl successively ₃Wash with EtOH.Organic moiety evaporation with being merged is adsorbed onto on the silica gel and with flash column chromatography it is carried out purifying (98: 2 CH ₂Cl ₂/ MeOH), thus the 1f of 790mg (2.89mmol, 58%) orange solids form obtained. ¹H?NMR(300MHz，CDCl ₃)δ10.01(s，1H)，8.64(d，J＝5.5Hz，1H)，8.09(br?S，1H)，7.96(d，J＝2.75Hz，1H)，7.37(d，J＝2.48Hz，1H)，7.29(d，J＝2.75Hz，1H)，7.08(dd，J＝2.47，5.5Hz，1H)，6.94(d，J＝9.35Hz，1H)，3.08(d，J＝5.23Hz，3H)。

Step 5

Imidazole ring forms (Baldwin, J.J.; Engelhardt, E.L.; Hirschmann, R.; Lundell, G.F.; Ponticello, G.S.J.Med.Chem 1979,22,687): with compound 1g (Lancaster (and Windham, NH), 25.75mL, (22.4g is 273mmol) at H 136.5mmol) to join NaOAc ₂Be heated to 100 ℃ of heating 40 minutes in the solution among the O (60mL) and with the solution of gained.After being cooled to room temperature (room temperature), the solution of 1h is joined 1f, and (25g is 91mmol) at NH ₄In the suspension in OH (150mL) and the methyl alcohol (450mL).The mixture of gained is at room temperature stirred a night.TLC (thin-layer chromatography, 95: 5 CH ₂Cl ₂/ MeOH) show that 1f is by completely consumed.This crude product is concentrated into aqueous slurries, and with it at saturated Na ₂CO ₃And CH ₂Cl ₂Between distribute.Water is used CH ₂Cl ₂Extract three times, and the organism that is merged is used brine wash, dry then (MgSO ₄) and it is concentrated, thereby obtain the 1i (83mmol) (yield is 91%) of 31.6g orange solids form.Do not need further to carry out purifying.Can prepare other midbody that is used to prepare substituted imidazoles in a similar manner.For example, midbody 1i ²Be that kind as shown below according to step 5 with 3,3,3-three fluoro-1-phenyl-propanes-1,2-two ketone hydrates replace 1h to come synthetic (MeOH=methyl alcohol, RT=room temperature, one night of o/n=, minute=minute):

Midbody 1i ³Be that kind as follows according to step 5 with 1-phenyl-1,2-propane diketone replaces 1b to carry out synthetic:

Midbody 1i ⁴Be that kind as follows according to step 5 with 1-(3-trifluoromethyl-phenyl)-1,2-propane diketone or 1-(4-trifluoromethyl)-1,2-propane diketone replace 1h to carry out synthetic:

Midbody 1i ⁵Be that kind as follows according to step 5, and combine US patent 5,374; Operation in 615 is used by 4,4; (2Z)-4,4 that 4-three fluoro-ethyl 3-oxobutanoates make, 4-three fluoro-2-(oxyimino)-ethyl 3-oxobutanoate replace 1h to carry out synthetic (NMA=N-methylacetamide):

Step 6

(45.76g, 120mmol) slurries in MeOH (220mL) and EtOAc (200mL) are used N with N-methyl-p-nitroaniline 1i ₂Sprayed 20 minutes, then to wherein adding 10%Pd/C (12.77g, 120mmol) suspension in MeOH (60mL).H is used in this reaction ₂Purify and with it at H ₂Kept under the atmosphere 2 days.This reaction is filtered with Celite pad and collected solid is washed with MeOH and EtOAc successively.With the organic filtrating evaporation that is merged, with the solid and the CH of gained ₂Cl ₂Azeotropic then with its dry night under vacuum, thereby obtains the 1j (yield is 96%) of 40.17g (115mmol) tawny powder type.LC/MS m/z 336.1 (MH ⁺), tR=1.81 minute.

Step 7

At room temperature, (23.37g, (40.17g is 115mmol) in the solution in MeOH (460mL) 115mmol) to join the diamines 1j that is carrying out stirring with isothiocyanic acid 4-trifluoromethyl ester.Should react and at room temperature keep 16 hours.After it reacts completely in judgement, in this reaction, add FeCl ₃(20.52g, 126.5mmol) solution in MeOH (50mL) and the mixture of gained at room temperature stirred a night.This reaction mixture bullion is joined in the separating funnel of a 3L who comprises EtOAc (750mL) and water (750mL).Carry out layer separation and water is extracted (preservation water) with EtOAc.Organic layer is merged, use saturated Na ₂CO ₃The aqueous solution, water and salt solution wash, and then it are carried out drying (MgSO ₄) and it is concentrated.Through adding saturated Na ₂CO ₃The aqueous solution makes the water alkalize (pH=10) preserved and the slurries of gained is joined in the separating funnel of a 3L who comprises EtOAc (500mL).Use filter paper filtering with this mixture stirring and with the emulsion of gained, carry out layer separation then and water is extracted (2 * 500mL) with EtOAc.Organic layer is merged, use brine wash, then it is carried out drying (MgSO ₄), concentrate in the extracting substance before joining and to it.The product that is merged is used CH ₂Cl ₂(500mL) grind, be adsorbed onto SiO ₂Upward and with flash column chromatography it is carried out purifying.Use CH at last ₂Cl ₂This material is ground, make { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine of pure white solid form.LC/MS m/z 519.1 (MH ⁺); ¹HNMR (300MHz, CDCl ₃) δ 8.44 (d, J=5.5Hz, 1H), 7.75 (d, J=8.8Hz, 2H), 7.61 (dd, J=2.2; 8.5Hz, 1H), 7.59 (d, J=8.8Hz, 2H), 7.56 (d, J=2.5Hz, 1H); 7.38 (app d, J=8.5Hz, 1H), 7.23 (d, J=1.9Hz, 1H), 6.96 (dd, J=2.2; 8.5Hz, 1H), 6.93 (dd, J=2.5,5.5Hz, 1H), 3.76 (s, 3H); LC/MS m/z=519.0, t _R=2.57 minutes (MH ⁺); C ₂₄H ₁₆F ₆N ₆The analytical calculation value of O: C 55.6, and H 3.11, N16.21; Measured value: C 55.81, H 3.43, and N 16.42; Mp:217-220 ℃.

Embodiment 2

The preparation of (2-fluoro-5-pyridin-3-yl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine

(2-fluoro-5-pyridin-3-yl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine is as the step 7 of top embodiment 1 is described, to carry out synthetic with 3-(4-fluoro-3-isocyanide sulfenyl-phenyl)-pyridine.LC/MS m/z 546.1 (MH ⁺), R _f1.82 minute.

Embodiment 3

The preparation of (2-fluoro-5-pyridin-4-yl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine

(2-fluoro-5-pyridin-4-yl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine is as the step 7 of top embodiment 1 is described, to carry out synthetic with 4-(4-fluoro-3-isocyanide sulfenyl-phenyl)-pyridine.LC/MS m/z 546.5 (MH ⁺), R _f1.83 minute.

Embodiment 4

The preparation of (4-tert-butyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine

(4-tert-butyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine is as the step 7 of top embodiment 1 is described, to carry out synthetic with 4-tert-butyl phenyl lsothiocyanates.LC/MS m/z 425.4 (MH ⁺), R _f2.56 minute.

Embodiment 5

{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(3-trifluoromethyl-phenyl)-amine

{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(3-trifluoromethyl-phenyl)-amine is as the step 7 of top embodiment 1 is described, to carry out synthetic with 3-(trifluoromethyl) phenyl lsothiocyanates.LC/MS m/z 519.4 (MH ⁺), R _f2.36 minute.

Embodiment 6

The preparation of (3-ethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine

(3-ethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl-oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine is as the step 7 of top embodiment 1 is described, to carry out synthetic with 3-ethylphenyl lsothiocyanates.LC/MS m/z 479.4 (MH ⁺), R _f2.32 minute.

Embodiment 7

The preparation of (4-chloro-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine

(4-chloro-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine is as the step 7 of top embodiment 1 is described, to carry out synthetic with 4-chloro-phenyl-lsothiocyanates.LC/MS m/z 485.4 (MH ⁺), R _f2.23 minute.

Embodiment 8

The preparation of (4-ethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine

(4-ethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl-oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine is as the step 7 of top embodiment 1 is described, to carry out synthetic with 4-ethylphenyl lsothiocyanates.LC/MS m/z 479.5 (MH ⁺), R _f2.31 minute.

Embodiment 9

The preparation of (4-chloro-3-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine

(4-chloro-3-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine is as the step 7 of top embodiment 1 is described, to carry out synthetic with 4-chloro-3-(trifluoromethyl) phenyl lsothiocyanates.LC/MS m/z 553.4 (MH ⁺), R _f2.51 minute.

Embodiment 10

The preparation of (4-fluoro-3-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine

(4-fluoro-3-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine is as the step 7 of top embodiment 1 is described, to carry out synthetic with 4-fluoro-3-(trifluoromethyl) phenyl lsothiocyanates.LC/MS m/z 537.4 (MH ⁺), R _f2.40 minute.

Embodiment 11

The preparation of { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethoxy-phenyl)-amine

{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(4-trifluoromethoxy-phenyl)-amine is as the step 7 of top embodiment 1 is described, to carry out synthetic with 4-(trifluoromethoxy) phenyl lsothiocyanates.LC/MS m/z 535.4 (MH ⁺), R _f2.24 minute.

Embodiment 12

(2-fluoro-5-trifluoromethyl-phenyl)-(1-methyl-5-{2-[5-methyl-4-(3-trifluoromethyl-phenyl)-1H-imidazoles-2-yl]-pyridin-4-yl oxygen base }-1H-benzimidazolyl-2 radicals-yl)-amine synthetic

(2-fluoro-5-trifluoromethyl-phenyl)-(1-methyl-5-{2-[5-methyl-4-(3-trifluoromethyl-phenyl)-1H-imidazoles-2-yl]-pyridin-4-yl oxygen base }-the 1H-benzimidazolyl-2 radicals-yl)-amine is to use and top embodiment 1 said similar operation, carries out synthetic with 2-fluoro-5-(trifluoromethyl) phenyl lsothiocyanates.LC/MSm/z 627.5 (MH ⁺), R _f2.79 minute.

Embodiment 13

(2-fluoro-5-trifluoromethyl-phenyl)-(1-methyl-5-{2-[5-methyl-4-(4-trifluoromethyl-phenyl)-1H-imidazoles-2-yl]-pyridin-4-yl oxygen base }-preparation of 1H-benzimidazolyl-2 radicals-yl)-amine

(2-fluoro-5-trifluoromethyl-phenyl)-(1-methyl-5-{2-[5-methyl-4-(4-trifluoromethyl-phenyl)-1H-imidazoles-2-yl]-pyridin-4-yl oxygen base }-the 1H-benzimidazolyl-2 radicals-yl)-amine is to use and top embodiment 1 said similar operation, carries out synthetic with 2-fluoro-5-(three-methyl fluoride) phenyl lsothiocyanates.LC/MSm/z 627.5 (MH ⁺), R _f2.79 minute.

Embodiment 14

2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-preparation of 5-Trifluoromethyl-1 H-imidazoles-4-ethyl formate

2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-5-Trifluoromethyl-1 H-imidazoles-4-ethyl formate is to use and top embodiment 1 said similar operation, carries out synthetic with 2-fluoro-5-(trifluoromethyl) phenyl lsothiocyanates.LC/MS?m/z609.5(MH ⁺)。

Embodiment 15

The preparation of (2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-5-Trifluoromethyl-1 H-imidazol-4 yl)-methyl alcohol

With Red-Al (hydrogenation two (2-methoxy ethoxy) aluminium sodium; 65% toluene solution; 0.1mL) be added drop-wise to 2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl-(0.0104g is 0.017mmol) in the solution in toluene for 5-Trifluoromethyl-1 H-imidazoles-4-ethyl formate.Observe effervesce and after 20 minutes, H is used in this reaction ₂O, NaOH extinguishing also extract it with EtOAc.Organic layer is used H ₂Na is used in the O washing ₂SO ₄Dry; Filter and it is concentrated; Thereby obtain 5.9mg (2-{4-[2-(2-fluoro-5-trifluoromethyl-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-5-Trifluoromethyl-1 H-imidazol-4 yl)-methyl alcohol bullion; Further use RP HPLC (reversed-phase HPLC) to carry out purifying it, thereby obtain the pure article (purity is 98%) of this compound of 1.1mg.LC/MS m/z 567.1 (MH ⁺), R _f2.40 minute.

Embodiment 16

2-{4-[1-methyl-2-(4-trifluoromethyl-phenyl amino)-1H-benzoglyoxaline-5-base oxygen base]-pyridine-2-yl }-preparation of 3H-imidazoles-4-formonitrile HCN

Will according to { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine of embodiment 1 preparation (1.83g, 3.4mmol) and 28%NH ₄The slurries of OH (23mL) in MeOH (10mL) are sealed in the test tube and are heated to 140 ℃ of heating 3 hours.After judging that with LC/MS it reacts completely, this reaction mixture bullion is joined in the separating funnel and with EtOAc (50mL) and water it distributed (50mL).Carry out layer separation and water is extracted (2 * 50mL) with EtOAc.Organic layer is merged, use brine wash, then it is carried out drying (MgSO ₄) and it is concentrated.This bullion is adsorbed onto SiO ₂Upward and with flash column chromatography it is carried out purifying, thereby obtain 2-{4-[1-methyl-2-(4-trifluoromethyl-phenyl amino)-1H-benzo-imidazoles-5-base oxygen base]-pyridine-2-yl of white solid form-3H-imidazoles-4-formonitrile HCN.LC/MS?m/z?476.1(MH ⁺)。

Embodiment 17-59a

Below the compound (embodiment 17-59a) shown in the table 1 be to prepare according to the operation described in the embodiment 1-16.Used various parent materials will be conspicuous (Tordeux for example, M. to those skilled in the art in these compounds synthetic; Langlois, B.; Wakselman, C.J Chem Soc.Perkin Trans 1 1990,2293).

Table 1

Embodiment 60

The preparation of (2-fluoro-5-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-pyridine-2-base-2H-[1,2,4] triazole-3-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine

The preparation of 4-[2-(2-fluoro-5-three fluoro-phenyl aminos)-1-methyl isophthalic acid H-benzoglyoxaline 5-base oxygen base]-pyridine-2-formonitrile HCN

Synthesizing of step 1.4-(4-amino-3-nitro-phenoxy) pyridine-2-formonitrile HCN:

With salt of wormwood (9g) vacuum-drying under the situation of heating, it is cooled to room temperature under nitrogen.Add 4-amino-3-nitro phenol (3.4g), 4-chloro-2-cyanopyridine (3.0g) and DMSO 99.8MIN. (30mL, anhydrous).This system is stirred under nitrogen, be heated to 103 ℃ and it was heated 1 hour simultaneously under this temperature.Then, this reaction cooled to room temperature, is poured into ice/H ₂On the O (500mL), collecting precipitation, washing (H ₂O), dissolving (EtOAc), dry (Na ₂SO ₄), filter, remove and desolvate, obtain a kind of solid.With its suspendible (Et ₂O), collect, air-dry 4.1g (73.5%) also collects second batch of product (0.55gm, 10%).m/z＝257(M+1)。

Step 2.N-[4-(2-cyanic acid-pyridin-4-yl oxygen base)-2-nitro-phenyl]-2,2,2-three fluoro-N-methyl-ethanamides synthetic:

With salt of wormwood (1.6g) vacuum-drying under the situation of heating, be cooled to room temperature and under nitrogen, it be suspended in the methylene dichloride (30mL) with 4-(4-amino-3-nitro-phenoxy) pyridine-2-formonitrile HCN (2.0g).It is cooled to 0 ℃ and to wherein adding purified trifluoroacetic anhydride (2.2mL).When adding, starting substance becomes solution rapidly.At 0 ℃ after following 10 minutes, this mixture is diluted washing (H with methylene dichloride ₂O, the NaCl aqueous solution), dry (K ₂CO ₃), filter, remove and desolvate, obtain a kind of yellow foam.m/z＝353(M+1)。This product is used under the situation of not carrying out purifying.Methyl iodide (0.53mL) is joined salt of wormwood (1.858g) under nitrogen (30mL, inclusion compound 2 is in～the suspension in 7.8mmol) at N DMF.This suspension is at room temperature stirred a night, then it is poured into H ₂Among the O (300mL), extraction (Et ₂O, 3 * 150mL), with the extract washing (H that is merged ₂O, the NaCl aqueous solution), dry (salt of wormwood) filters, and removes and desolvates, and obtains a kind of orange oily matter (7.4922g).m/z＝367(M+1)。

Synthesizing of step 3.4-(4-methylamino-3-nitro-phenoxy)-pyridine-2-formonitrile HCN:

At room temperature, NaOH (1mL, 1N aq.) is added drop-wise to N-[4-(2-cyanic acid-pyridin-4-yl oxygen base)-2-nitro-phenyl]-2,2, and 2-three fluoro-N-methyl-ethanamides (3,440mg) in the solution in ethanol (6mL).After 40 minutes, this mixture is used H ₂O (20mL) dilutes and it is cooled to 0 ℃.Collect the crystal of bright orange, washing (H ₂O) and with its air-dry 311.1mg (94%).m/z＝271(M+1)

Synthesizing of step 4.4-[2-(2-fluoro-5-three fluoro-phenyl aminos)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-formonitrile HCN:

Palladium is draped over one's shoulders carbon, and (46mg 10%w/w) is suspended under nitrogen among the MeOH (2mL).Under nitrogen, at room temperature, the suspension of gained is joined in 4-(4-methylamino-3-nitro-phenoxy)-pyridine-suspension of 2-formonitrile HCN (311mg) in MeOH (3mL).Change its atmosphere into hydrogen, and with this system vigorous stirring 1 hour under 1atm hydrogen.Then, change its atmosphere into nitrogen, this mixture filtration (zeyssatite) also will be filtrated do not carry out being used for next reaction with being further purified.m/z＝242(M+1)。2-fluoro-5-trifluoromethyl lsothiocyanates (250mg) is joined in the solution of compound 5 in MeOH (10mL).This solution was stirred 2 hours under refluxing.After it reacts completely in judgement, in this reaction, add anhydrous FeCl ₃(1.3 eq.5 244mg) also at room temperature stirs a night with the mixture of gained.This reaction mixture bullion is joined in the separating funnel that comprises EtOAc and water.Carry out that layer separates and water is extracted with EtOAc.Organic layer is merged, use saturated Na ₂CO ₃The aqueous solution, water and salt solution wash, and then it are carried out drying (MgSO ₄) and it is concentrated.This material is handled (on silica gel, with the dichloromethane solution gradient elution of 0-5%MeOH) with chromatogram, and the yield in 28% (with compound 4) separation obtains required compound.m/z＝428(M+1)。

Step 5. (2-fluoro-5-trifluoromethyl-phenyl)-{ 1-methyl-5-[2-(5-pyridine-2-base-2H-[1,2,4] triazole-3-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-amine

Step 6

Be dissolved among the EtOH (0.1M) 4-[2-(4-fluoro-phenyl amino)-1-methyl isophthalic acid H-benzoglyoxaline-5-base oxygen base]-pyridine-2-formonitrile HCN and to wherein adding NaOEt (1eq.; 0.5M EtOH solution), then to wherein adding pieolinyl hydrazides (1eq.) and this solution being heated 2000 seconds down at 140 ℃ in microwave.Then, this reaction mixture is concentrated and with reversed-phase HPLC it carried out purifying, thereby obtain required product, m/z=547 (M+1).

Embodiment 61-64

Below the compound (embodiment 61-64) shown in the table 2 prepare according to embodiment 60 described operations.

Table 2

Embodiment 65

The preparation of N-(4-hydroxyl-2-nitrophenyl)-methane amide

N-(4-hydroxyl-2-nitrophenyl)-methane amide can prepare according to following method:

1. set up 3-L, a 5-neck reaction flask of being furnished with internal temperature probe, temperature regulator, heating jacket, condensing surface, mechanical stirrer, 1-L feed hopper and nitrogen inlet.With this reactor drum with nitrogen flush 5 minutes.

2. in this flask, add diacetyl oxide (245mL).It is stirred under nitrogen.

3. disposable adding formic acid (125mL) (because heat release is observed in the reaction between mixing and diacetyl oxide and the formic acid).

4. internal temperature (IT) terminal point is arranged to 60 ℃ and begin to heat.After its IT reaches 60 ℃, stir and it was kept 2 hours again.

5. with ice bath inclusion is cooled off.

6. when its IT reaches envrionment temperature (about 20 ℃), the feed hopper through 1-L begins to add the solution of 4-amino-3-nitro phenol (160g) in the anhydrous THF of 700mL (THF) in batches, makes its IT be no more than 40 ℃.Product begins to be precipitated out with the yellow solid form.

7. ought add fashionablely fully, replace ice bath with heating jacket.The IT terminal point is arranged on 60 ℃ and begin to heat.

8. with HPLC reaction process is monitored.This reaction is carried out less than 1 hour usually.

9. when the area percentage of parent material＜1%, add 500mL water.It is cooled to room temperature with ice bath.

10. collect product through vacuum filtration.Filter cake is washed with 3 * 200mL water.Air-dry, and it is further dry at the air of under the 27in.Hg vacuum, using gentleness under 50 ℃ or nitrogen gas stream in baking oven, until reaching constant weight.

Embodiment 66

The preparation of 4-methylamino-3-nitrophenols

4-methylamino-3-nitrophenols can prepare according to following operation:

1. set up 500mL, a 3-neck reaction flask of being furnished with internal temperature probe and nitrogen inlet.With this reactor drum with nitrogen flush 5 minutes.

2. in this reactor drum, add N-(4-hydroxyl-2-nitrophenyl)-methane amide (5g) and anhydrous THF (100mL).With it at N ₂Under stir, thereby obtain a kind of xanchromatic slurries.

3. slowly add boron trifluoride diethyl ether compound (3.83mL) through syringe.

4. this reaction mixture was at room temperature stirred 30 minutes.

Through feed hopper to wherein add Peng Qinghuana (1.04g) in batches.

6. should react stirring 1 hour and per hour (reaction needs 3 hours usually) monitored in this reaction thereafter with HPLC.

7. when the HPLC sample shows that parent material is lower than 1.0%, in 10 minutes, slowly add 1M HCl (40mL) through syringe.

8. stirred 60 minutes.

As required through syringe to wherein adding 1M NaOH so that its pH is transferred to 7 ± 0.5.

10. be poured into this reaction mixture in the round-bottomed flask of a 500mL and with its concentrating under reduced pressure (20mm Hg, 25 ℃) until removing the clarifying liquid of about 100mL.

11. in this reaction vessel, add entry (100mL).Under condition of stirring, it is cooled to 0 ± 2 ℃.Product is precipitated out with the form of red solid.

12. collect product with vacuum filtration through coarse sintered filter funnel.The filter cake water is washed (2 * 20mL).Air-dry, then that it is dry until constant weight under 50 ℃/27in.Hg in baking oven.Sampling is analyzed.

Embodiment 67

The preparation of 4-chloropyridine-2-carbonyl chlorine

4-chloropyridine-2-carbonyl chlorine can prepare according to following operation:

1. set up the pneumatic outlet that is positioned at condenser overhead being furnished with internal temperature (IT) probe, temperature regulator, heating jacket, condensing surface, mechanical stirrer, nitrogen inlet, linking to each other with the 2-neck liquid trap (it links to each other with a washer of having filled the 12L of about 6 liters of 8M NaOH solution again) of a 2L 5-L 5-neck reaction flask and with magnetic stirring apparatus it is stirred.This reactor drum with nitrogen flush 5 minutes, is cut off nitrogen gas stream then.

2. in this reactor drum, add THIONYL CHLORIDE 97 (1.18L),, keep appropriateness simultaneously and stir (about 200rpm) then to wherein adding Potassium Bromide (38.4g).

3. in this reactor drum, add Picolinic Acid (397g).

4. its IT terminal point is set in 80 ℃ and begin to heat.

5. sampling and reaction process is monitored with HPLC.This reaction is carried out needing about 14 hours fully usually.Prolong heating and will cause more dichloride effect.

6. when thinking that reaction finishes, (in this reaction mixture, exist and be lower than 1% VPP), stop heating.Remove heating jacket.

7. when its IT is lower than 30 ℃, this liquid is transferred in the reaction flask of a 3-L.Clean with the reactor drum of 700mL toluene this 5-L.Scavenging solution is transferred in the flask of 3-L.Under reduced pressure remove excessive SOCl ₂And toluene.Repeat this process with 2 * 700mL toluene.Remove all solvents, obtain a kind of yellow-orange solid.In this reaction mixture, add toluene (400mL).Mixture with gained carries out next step.

Embodiment 68

The preparation of 4-chloropyridine-2-formic acid uncle-butyl ester

4-chloropyridine-2-formic acid uncle-butyl ester can prepare according to following operation:

1. go up make-up machinery whisking appliance and TM at the round-bottomed flask (the 4-neck) of a 12L.

2. in this reactor drum, add toluene (1L), pyridine (977.7g) and tert-Butyl dicarbonate (BOC) ₂O (855.5g).

3. with this reactor cooling, it is 0 ℃ thereby make its internal temperature.

4. so that being lower than 5 ℃ speed, the internal temperature of this reaction in this reactor drum, adds 4-chloropyridine-2-carbonyl chlorine (686g).

5. make this sluggish be warming up to room temperature (～20 ℃) and it was stirred 16 hours.

6. when confirming with HPLC should reaction to finish (parent material＜0.5 area %), with this reaction solution with water washing (2 * 4L), use 1M HCl solution washing (2 * 2L) then.

With this reaction mixture concentrating under reduced pressure to remove toluene and remaining pyridine.

8. add toluene (500mL),, thereby obtain required product then with this reaction mixture concentrating under reduced pressure.

Embodiment 69

The preparation of 4-(4-methylamino-3-nitro-phenoxy)-pyridine-2-t-butyl formate

4-(4-methylamino-3-nitro-phenoxy)-pyridine-2-t-butyl formate can prepare according to following operation:

1. make-up machinery whisking appliance, TM and nitrogen inlet on the round-bottomed flask of a 3L.

2. in this reactor drum, add K ₂CO ₃(123g).

3. this reaction vessel is placed under the inert atmosphere.

4. in this reactor drum, add 4-methylamino-3-nitrophenols (100g), 4-chloropyridine-2-t-butyl formate (127g) and anhydrous DMSO (1L).

5. should react vigorous stirring and be heated to 100 ℃.

6. when confirming through HPLC should reaction to finish (＜0.5 area %4-chloropyridine-2-t-butyl formate), this thermal reaction mixture is poured into 3L carrying out in the cold water (volume) that stirs.

7. through filtering to isolate orange required product to orange-brown solid.

8. (2 * 200mL) use heptane then, and (2 * 200mL) wash with the solid water of separating.

9. this material is dried to constant weight in vacuum drying oven under 45-50 ℃.

Embodiment 70

The preparation of 4-(4-(methylamino)-3-nitro-phenoxy) pyridine-2-formaldehyde

4-(4-(methylamino)-3-nitro-phenoxy) pyridine-2-formaldehyde can prepare according to following operation:

1. on the round-bottomed flask of a 1000mL, assemble nitrogen inlet, mechanical stirrer and TM.

2. in this reactor drum, add 4-(4-methylamino-3-nitro-phenoxy)-pyridine-2-t-butyl formate (10g) through powder funnel.

3. add 2-methyl THF (100mL) through powder funnel.

4. it is-25 ℃ with this reactor cooling to internal temperature.

5. add DIBAL (diisobutylaluminium hydride, the toluene solution of 1.5M with the speed that its internal temperature is remained on-15 ℃ through feed hopper; 72mL).

6. through HPLC or GC (gc) this reaction is analyzed, whether the inspection ester disappears.

7. this is reflected under-20 ℃ and stirs, per hour it is monitored.

8. if do not make progress after this is reflected at 2 hours, again to wherein adding 0.5 equivalent DIBAL (diisobutylaluminium hydride) and this reaction being monitored.Repeating this step all is consumed until all esters.

9. in case should reaction finish, with MeOH (10mL) with its slow extinguishing.

10. sodium-potassium tartrate (40g) is joined in the 200mL water and also stir so that its dissolving.

11. this aqueous solution joined in the reaction mixture and makes it heat to room temperature.

12. in this reaction vessel, add 2-methyl THF (100mL).

13. under condition of stirring, this reaction is heated to 50 ℃ of heating 1 hour.

14. it is separated.

15. remove following water layer.

16. organic layer is filtered with Celite pad.

17. (2 * 50mL) clean this zeyssatite with 2-methyl THF.

18. this reaction mixture is joined in the round-bottomed flask of a 500mL.

19. this reaction mixture is concentrated into～50mL through distillation.

20. under condition of stirring, this reaction mixture is cooled to 0 ℃.

21. this reaction mixture was stirred 1 hour down at 0 ℃.

22. this reaction mixture is filtered with coarse incipient fusion filter.

23. make these solids on this filter dry 30 minutes to 1 hour.

24. these solids are analyzed to measure pure %, remove pure impurity if desired, then with its 1 hour (5mL methyl alcohol/g compound) of under 30 ℃, in methyl alcohol, pulling an oar with GC and NMR.

Embodiment 71

The preparation of 4-(2-(5-(trifluoromethyl)-1H-imidazoles-2-yl)-pyridin-4-yl oxygen base)-N-methyl-2-N-methyl-p-nitroaniline

4-(2-(5-(trifluoromethyl)-1H-imidazoles-2-yl) pyridin-4-yl oxygen base)-N-methyl-2-N-methyl-p-nitroaniline can prepare according to following operation:

1. go up make-up machinery whisking appliance, internal temperature probe, temperature regulator and condensing surface at the round-bottomed flask (3 necks) of a 2L.

2. in this reactor drum, add entry (590mL) through powder funnel.

3. begin this mixture is stirred and in this reactor drum, adds sodium acetate (240g).

4. water (30mL) cleans the flask that is used to adorn sodium acetate.

5. this reaction is heated to 50 ℃.

6. under 50 ℃, add 3 in batches, 3-two bromo-1,1,1-trifluoropropyl-2-ketone (395g), the internal temperature that will react simultaneously remains under 100 ℃.

7. this reaction is heated to 100 ℃ internal temperature.

8. stirring is after 1 hour down this being reflected at 100 ℃, and sampling is analyzed.

9. this is reflected at and is stirred to parent material＜1.5% under 100 ℃.

10. in a single day should reaction finish, this reaction mixture is cooled to＜65 ℃.

11. this reaction is being carried out refrigerative simultaneously, is being gone up assembling internal temperature probe, temperature regulator, reflux exchanger and mechanical stirrer at the round-bottomed flask (the 4 neck flasks that chuck is arranged) of a 5L.

12. in the reactor drum of a 5L, add ETHYLE ACETATE (500mL) and begin to stir through powder funnel.

13. in the reactor drum of a 5L, add 4-(4-(methylamino)-3-nitro-phenoxy) pyridine-2-formaldehyde (200g) through powder funnel.

14. wash this powder funnel and washing fluid be added in the reactor drum of 5L with ETHYLE ACETATE (200mL).

15. the ethanol (1.3L) of adding 95% in the reactor drum of this 5L.

16. this pyruvic aldehyde reaction mixture is transferred to the reactor drum of 5L from the reactor drum of 2L.At this moment the temperature of this mixture is～35 ℃.

17. slowly add dense NH in batches ₄OH (1.3L) monitors its temperature simultaneously.This reaction is heat release, and therefore initial 500mL should add when its internal temperature being remained on 50 ℃ times in batches.Total joining day is～25 minutes.The temperature that raises makes end product become redness.

18. the reactor drum of this 5L is heated to 50 ℃.

19. this reaction mixture is stirred under 50 ℃.At this moment, the color of this solution be generally slightly redly-orange.

React completely 20. per hour all this reaction is monitored until it.

21. when thinking that it reacts completely, this reaction mixture is cooled to 0 ℃ of cooling 2 hours.

22. come the filtering separation product through coarse sintered glass filter.

23. (150mL) cleans this reactor drum with cold ethanol.This scavenging solution is transferred in the filter.

24. in the reactor drum of a 5L, add entry (2L).

25. stir and with this reactor cooling to 10 ℃.

26. this wet cake is transferred to the reactor drum of this 5L from filter.

27. it was stirred 60 minutes down at 10 ℃.

28. product is leached through coarse sintered glass filter.

29. water (250mL) cleans this reactor drum.This scavenging solution is transferred in the filter.

30. with wet cake on filter dry 1 hour.

31. product is transferred in the round-bottomed flask (single neck) of a 2L and with rotatory evaporator its upset is dried to and record constant weight with 45 ℃ bath temperature.

Embodiment 72

4-(2-(5-(trifluoromethyl)-1H-imidazoles-2-yl) pyridin-4-yl oxygen base)-N1-methylbenzene-1, the preparation of 2-diamines

4-(2-(5-(trifluoromethyl)-1H-imidazoles-2-yl) pyridin-4-yl oxygen base)-N1-methylbenzene-1, the 2-diamines can prepare according to following operation:

1. go up make-up machinery whisking appliance, internal temperature probe, temperature regulator, nitrogen purge and reflux exchanger at the round-bottomed flask (4 neck) of a 2L.

2. in this reactor drum, add EtOH (125mL) through powder funnel.Begin rapidly to stir.

3. in this reactor drum, add 4-(2-(5-(trifluoromethyl)-1H-imidazoles-2-yl) pyridin-4-yl oxygen base)-N-methyl-2-N-methyl-p-nitroaniline (50g) through powder funnel.

4. this reaction is heated to 50 ℃.

5. when this reaction is heated, in the Erlenmeyer flask of a 250mL, add entry (75mL) through powder funnel.Begin rapidly to stir.

6. in the Erlenmeyer flask of this 250mL, add 3.0eq. yellow soda ash (41.92g) through powder funnel.

7. this mixture being stirred to all solids all dissolves.

8. in case this suspension reaches 50 ℃, through powder funnel this sodium carbonate mixture is transferred to the reaction mixture from the Erlenmeyer flask of 250mL.

9. in the Erlenmeyer flask of a 250mL, add entry (75mL) through powder funnel.Begin rapidly to stir.

Face join in this reaction flask before, in the Erlenmeyer flask of this 250mL, add 1.0eq. V-Brite B (22.95g) through powder funnel.

11. this solid is stirred rapidly, until its most of dissolving.

12. this V-Brite B mixture is transferred to the reaction mixture rapidly from the Erlenmeyer flask of 250mL through powder funnel.

13. this is reflected at 50 ℃ stirred 30 minutes down.

14. in the Erlenmeyer flask of a 250mL, add entry (75mL) through powder funnel.Begin rapidly to stir.

15. face join in this reaction flask before, in the Erlenmeyer flask of this 250mL, add 1.0eq. V-Brite B (22.95g) through powder funnel.

16. this solid is stirred rapidly, until its most of dissolving.

17. this V-Brite B mixture is transferred to the reaction mixture from the Erlenmeyer flask of this 250mL rapidly through powder funnel.

18. this is reflected at 50 ℃ stirred 30 minutes down.

19. in the Erlenmeyer flask of a 250mL, add entry (150mL) through powder funnel.

20. face join in this reaction flask before, in the Erlenmeyer flask of this 250mL, add 2.0eq. V-Brite B (45.90g) through powder funnel.

21. this solid is stirred rapidly, until its most of dissolving.

22. this V-Brite B mixture is transferred to this reaction mixture from the Erlenmeyer flask of this 250mL rapidly through powder funnel.

23. this is reflected at 50 ℃ stirred 60 minutes down.

24. sampling is to confirm that it reacts completely.

25. if completeness＞98% that should react then carry out step 36.If no, then continue step 26.

26. in the reaction flask of a 2L, add 1.0eq. V-Brite B (22.95g) through powder funnel.

27. this reaction mixture was stirred 60 minutes rapidly down at 50 ℃.

28. sampling is to confirm that it reacts completely.

29. if completeness＞98% that should react then carry out step 36.If no, then continue step 30.

30. in the reaction flask of a 2L, add 1.0eq. yellow soda ash (13.97g) through powder funnel.

31. this reaction mixture was stirred 15 minutes rapidly down at 50 ℃.

32. in the reaction flask of a 2L, add 1.0eq. V-Brite B (22.95g) through powder funnel.

33. this reaction mixture was stirred 60 minutes rapidly down at 50 ℃.

34. sampling is to confirm that it reacts completely.

35. when the completeness of this reaction＞98%, carry out step 36.

36. when thinking that it reacts completely, in the reaction flask of a 2L, add entry (125mL) through powder funnel.

37. this reaction mixture is cooled to 10 ℃ and it was stirred 1 hour.

38. product is separated through filtering with coarse sintered glass filter.

39. water (50mL) cleans this reactor drum.This scavenging solution is transferred in the filter.

40. wet cake is dried to it on this filter no longer drip.

41. in the reaction flask of a 2L, add entry (500mL) through powder funnel.

42. this filter cake is shifted back in the reaction flask through powder funnel.

43. this material was at room temperature stirred 60 minutes.

44. product is separated through filtering with coarse sintered glass filter.

45. water (25mL) cleans this reactor drum.This scavenging solution is transferred in the filter.

46. with wet cake on filter dry about 1 hour.

47. product is transferred in the 2L round-bottomed flask (single neck) and with 50 ℃ bath temperature, with rotatory evaporator with its drying of slowly overturning until recording constant weight.

Embodiment 73

{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzo-imidazoles-2-yl }-(4-trifluoromethyl-phenyl)-amine can prepare according to following operation:

1. make-up machinery whisking appliance, internal temperature probe, temperature regulator, nitrogen purge and condensing surface on the round-bottomed flask of a 2-L, 4-neck.

2. in this reactor drum, add 4-(2-(5-(trifluoromethyl)-1H-imidazoles-2-yl) pyridin-4-yl oxygen base)-N1-methylbenzene-1,2-diamines (200g) through powder funnel.

3. in this reactor drum, add acetonitrile (1L) through powder funnel.

4. beginning is stirred this mixture under envrionment temperature and nitrogen atmosphere.

5. after 20 ± 5 minutes, in this reactor drum, add 4-trifluoromethyl lsothiocyanates (104g) through powder funnel.

After adding lsothiocyanates sampling in 30 minutes to confirm that it reacts completely.

7., this mixture is filtered with coarse sintered glass filter in case it reacts completely.

8. with MeCN (200mL) this reactor drum is cleaned.This scavenging solution is transferred in the filter.

9. with MeCN (200mL) solid that takes out is washed.

10. filtrating is transferred in the round-bottomed flask of a 3-L who is furnished with mechanical stirrer, internal temperature probe, temperature regulator, nitrogen purge and condensing surface, 4-neck.

11. in this reactor drum, add N through powder funnel, the N-diisopropyl ethyl amine.

12. every at a distance from 10 minutes the branch quarter (time of interpolation always is 30 minutes) pass through powder funnel and in this reactor drum, add 2-chloro-1,3-methylimidazole quinoline

muriate.After adding the last time, with this reaction mixture restir 10 minutes.

13. this reaction is heated to 50 ℃ ± 5 ℃.

14. this mixture heating up is being taken a sample after 30 minutes to confirm that it reacts completely.

15., this reaction mixture is transferred in the 3-L round-bottomed flask of an assembling as in the step 10 through a kind of 0.2 online μ M membranous filter in case it reacts completely.

16. add entry through powder funnel.

17. this reaction is heated to 50 ℃ ± 5 ℃.

18. after heating 2 hours, make this reaction mixture be cooled to 20-25 ℃ and with its restir 1 hour.

19. product is separated through filtering with medium sintered glass filter.

20. the MeCN/ water (300mL) with 2: 1 cleans this reactor drum.This scavenging solution is transferred in the filter.

21. the MeCN/ water (300mL) with 2: 1 washs filter cake.

22. with wet cake on filter dry about 1 hour.

23. product transferred on the drying basin and this material is dried to remaining MeCN (acetonitrile) with small nitrogen gas stream down at 70 ± 5 ℃ in vacuum drying oven measure and be lower than 410ppm.

24., product is being furnished with in the EtOH of 15 times of volumes (weight/volume) in the reactor drum of mechanical stirrer, internal temperature probe, temperature regulator, nitrogen purge and condensing surface and is being heated to backflow for recrystallization.

25. when replacing condensing surface, this mixture was refluxed 30 minutes with still head.

26. distillation EtOH is until residue 4 volumes.Stop heating and add 1 water by volume.

27. make this mixture be cooled to 0-5 ℃.

28. product is separated through filtering with medium sintered glass filter.

29. this reactor drum is cleaned with 4: 1 EtOH/ water (1 volume).This scavenging solution is transferred in the filter.

30. water (1 volume) cleans filter cake.

31. with wet cake on filter dry about 1 hour.

32. product transferred in the drying basin and this material is dried to small nitrogen gas stream down at 50 ℃ ± ℃ in vacuum drying oven reach constant weight.

Embodiment 74

(200mg 1mmol) joins 4-(2-(5-(trifluoromethyl)-1H-imidazoles-2-yl) pyridin-4-yl oxygen base)-N1-methylbenzene-1, and (350mg is 1mmol) in the mixture in the 3mL acetonitrile for the 2-diamines with 4-trifluoromethyl lsothiocyanates.After it was stirred 20 minutes at ambient temperature, it transformed the HPLC analysis revealed fully.Add triethylamine (0.3mL; 2.2mmol); Add then 2-chloro-1-picoline

iodide (270mg, 1.05mmol).This reaction mixture is heated to 50 ℃ of heating 5 hours.Stop heating and add 1.5mL water.This mixture was being stirred 2 hours, through solid collected by filtration and with 2: 1 acetonitrile/water to its wash (3 * 1mL), thus obtain 317mg (61%) title compound.

Embodiment 74a

The preparation of 4-(2-(5-(trifluoromethyl)-1H-imidazoles-2-yl) pyridin-4-yl oxygen base)-N-methyl-2-N-methyl-p-nitroaniline

NaOMe (1.5mL, 6.3mmol, the MeOH solution of 25 weight %) is joined 4-(4-(methylamino)-3-nitro-phenoxy) pyridine-2-formonitrile HCN, and (1.72g is 6.3mmol) in the mixture in 1-PrOH (10mL).With this mixture heating up to 50 ℃ (internal temperature).After being heated 1 hour, HPLC analysis revealed parent material transforms fully.Add NH ₄(1.46g is 18.9mmol) and with this mixture heating up to 70 ℃ for OAc.With it 70 ℃ of down heating after 1 hour, with this mixture heating up to 85 ℃.Simultaneously, whenever divided 4 parts to add 3-bromo-1,1 with the amount of every part of 0.2-mL at a distance from 30 minutes, and the 1-trifluoroacetone (0.8mL, 7.56mmol).This mixture was heated 20 hours down at 85 ℃.Then, make this mixture be cooled to envrionment temperature and add entry.After stirred for several hour, this mixture is cooled off in ice/water-bath.It was cooled off 1 hour in ice/water-bath after, (2 * 7mL) washed it through solid collected by filtration and with 1: 1 1-PrOH/ water.This solid is descended dry about 16 hours at 50 ℃ in vacuum drying oven, thereby obtain 0.982g (41%) title compound.

Embodiment 74b

The preparation of 4-chloro-2-(5-(trifluoromethyl)-1H-imidazoles-2-yl) pyridine

NaOMe (0.46mL, 2mmol, the MeOH solution of 25 weight %) is joined 4-chloro-2-cyanic acid-pyridine, and (277mg is 2mmol) in the mixture in 1-PrOH (3mL).With this mixture heating up to 50 ℃ (reaction-blocking-up temperature).After heating 1 hour, HPLC analysis revealed parent material transforms fully.With this mixture heating up to 70 ℃ and add NH ₄OAc (462mg, 6mmol).At 70 ℃ after following 1 hour, with this mixture heating up to 85 ℃.Simultaneously, whenever divided 4 parts to add 3-bromo-1,1 with the amount of every part of 0.063mL at a distance from 30 minutes, and the 1-trifluoroacetone (0.25mL, 2.4mmol).This mixture was heated about 20 hours down at 85 ℃.HPLC shows that this this bullion is 72.4% (LCAP) and analyzes with LC-MS it is confirmed.

Embodiment 74c

4-chloro-2-cyanic acid-pyridine

Cooling unit through the outside is cooled to 0.2 ℃ with 4-chloro-2-pyridine carboxamide (93.9g, 0.6 mole) and TEA (125mL, 0.9 mole) in EtOAc (500mL).In 40 minutes, add TFAA (92mL, 0.66 mole) through feed hopper.During adding, its internal temperature rises to 10 ℃.Adding fashionablely fully, its temperature is 0.0 ℃.After adding, turn off water cooler.After having spent 30 minutes again, the remaining parent material of HPLC analysis revealed is 4.3% (LCAP).Again to wherein adding 8.3mL (0.06 mole) TFAA.With this reaction mixture restir after 20 minutes, it transforms the HPLC analysis revealed fully.Add 10%K ₂CO ₃The aqueous solution (w/v, 500mL).Its internal temperature rises to 22.0 ℃ from 13.7.After stirring 20 minutes, this mixture is transferred in the separating funnel.Carry out layer separation and water layer is extracted (150mL) with EtOAc.(w/v 300mL) washs, dry (Na with 10% aqueous citric acid solution with the organic layer that is merged ₂SO ₄), filter and concentrate.This bullion is descended dry 16 hours at 50 ℃ in vacuum drying oven, thereby obtains 72.85g (87%) title compound: ¹H NMR (400MHz, CDCl ₃) δ 8.6 (m, 1H), 7.7 (m, 1H), 7.5 (m, 1H); ¹³C NMR (100MHz, CDCl ₃) δ 151.8,145.3,134.9,128.7,127.4,116.1; HPLC＞99% (LCAP).

Embodiment 74d

4-(4-methylamino-3-nitro-phenoxy)-pyridine-2-formonitrile HCN

With 4-chloro-2-cyanic acid-pyridine (6.9g, 0.05 mole), 4-methylamino-3-nitrophenols (8.4g, 0.05 mole) and K ₂CO ₃(10.4g, 0.075 mole) mixture heating up to 60 in DMSO (80mL) ℃.After 11.5 hours, two kinds of parent materials of HPLC analysis revealed all transform fully.After being cooled to 20 ℃, in this reaction mixture, add entry (240mL).Its temperature rises to 40 ℃, reduces to envrionment temperature then.Through solid collected by filtration and water it is washed (2 * 40mL).Then, this solid is formed slurries in heptane (40mL).Collecting solid also washs it with heptane (40mL).This bullion is descended dry 16 hours at 50 ℃ in vacuum drying oven, thereby obtains 10.33g (76%) title compound: ¹H NMR (400MHz, DMSO-d ₆) δ 8.5 (m, 1H), 8.2 (m, 1H), 7.9 (m, 1H), 7.7 (m, 1H), 7.5 (m, 1H), 7.2 (m, 1H), 7.1 (m, 1H), 3.0 (s, 3H); ¹³C NMR (100MHz, DMSO-d ₆) δ 165.1,152.9,144.4,140.6,134.1,130.4,130.1,117.9,117.1,117.0,116.5,114.9,29.8; APCI MS [M+H] ⁺=271; HPLC＞99% (LCAP).

Embodiment 74e

4-(4-methylamino-3-amino-phenoxy)-pyridine-2-formonitrile HCN

4-(4-methylamino-3-nitro-phenoxy)-pyridine-2-formonitrile HCN (5.0g, 0.019 mole) is heated to 40 ℃ in EtOH (15mL).Add Na ₂CO ₃(4.7g, 0.044 mole) adds H then ₂O (8.4mL).Add Na ₂S ₂O ₄(3.3g, 0.019 mole) adds H then ₂O (10mL).Its temperature rises to 49.5 ℃ from 41.7.After being cooled to 41.7 ℃, add Na ₂S ₂O ₄(3.3g, 0.019 mole) adds H then ₂O (10mL).Its temperature rises to 44.5 ℃.After being cooled to 36.7 ℃, add Na ₂S ₂O ₄(6.6g, 0.038 mole) adds H then ₂O (20mL).Its temperature rises to 44.0 ℃.HPLC analysis revealed parent material is 4.1% (LCAP).Again to wherein adding some Na ₂S ₂O ₄(3.3g, 0.019 mole).Its restir after 15 minutes, is being removed heating and added H ₂O (12.5mL).Under 25 ℃, add some Na again ₂CO ₃(1.3g, 0.012 mole) also cools off this mixture in ice/water-bath.When being lower than 5 ℃, make this mixture ageing 30 minutes (final temperature is 1.5 ℃).Through solid collected by filtration and use H ₂O (10mL, 5mL then) washs it.With this solid on filter dry 30 minutes, transfer to it in reaction flask then and to wherein adding H ₂O (50mL).This mixture was stirred 45 minutes.Then, through solid collected by filtration and use H ₂O washs (2 * 10mL) to it.This bullion is descended dry 16 hours at 50 ℃ in vacuum drying oven, thereby obtains 3.50g (76%) title compound: ¹H NMR (400MHz, DMSO-d ₆) δ 8.5 (m, 1H), 7.5 (m, 1H), 7.1 (m, 1H), 6.4 (m, 1H), 6.3 (m, 2H), 4.8 (s, 2H), 4.7 (s, 1H), 2.7 (s, 3H); APCI MS [M+H] ⁺=241; HPLC＞99% (LCAP).

Embodiment 74f

4-[1-methyl-2-(4-(trifluoromethyl) phenyl amino)-1H-benzoglyoxaline-5-base oxygen base]-pyridine-2-formonitrile HCN

4-(trifluoromethyl) phenyl lsothiocyanates (9.65g, 0.0475 mole) is joined in 4-(4-methylamino-3-amino-phenoxy)-solution of pyridine-2-formonitrile HCN (12.0g, 0.05 mole) in MeCN (60mL).After 40 minutes, this amine of HPLC analysis revealed transforms fully.(2 * 12mL) wash with MeCN with this mixture filtration and with the solid that takes out.In filtrating, add DIPEA (17.5mL, 0.1 mole).Whenever divided 4 parts to add 2-chloro-1,3-methylimidazole quinoline

muriate (DMC) at a distance from 10 minutes with the amount of every part of 2.11g.After adding the last time, with this mixture restir 10 minutes, at this moment the HPLC analysis revealed its transform fully.Then, with this mixture heating up to 50 ℃ (internal temperature).At 50 ℃ after following 45 minutes, it changes into product fully the HPLC analysis revealed.This mixture is cooled to envrionment temperature, then to wherein adding H ₂O (45mL).This reaction mixture begins to become evenly, and compound begins from this mixture, to be precipitated out then.After it is stirred 2 hours, through solid collected by filtration and with 2: 1 MeCN/H ₂(2 * 20mL) wash it O.This bullion is descended dry 16 hours at 50 ℃ in vacuum drying oven, thereby obtain 16.10g (78%) title compound. ¹H?NMR(400MHz，DMSO-d ₆)δ9.5(m，1H)，8.5(m，1H)，8.0(m，2H)，7.7(m，2H)，7.6(m，1H)，7.4(m，1H)，7.3(m，1H)，7.1(m，1H)，6.9(m，1H)，3.7(m，3H)；APCI?MS[M+H] ⁺＝410；HPLC＞99％(LCAP)。

Embodiment 74g

{ 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine

NaOMe (0.23mL, 1mmol, the MeOH solution of 25 weight %) is joined embodiment 74f, and (409mg is 1mmol) in the mixture in MeOH (4mL).After 1 hour, HPLC analysis revealed parent material is 46.2% (LCAP) at ambient temperature.With this mixture heating up to 50 ℃ (reaction-blocking-up temperature).After heating 1 hour, the remaining parent material of HPLC analysis revealed is 4.1% (LCAP).Add NH ₄OAc (231mg 3mmol), adds 3-bromo-1,1 then, and the 1-trifluoroacetone (0.13mL, 1.2mmol).This mixture was heated about 20 hours down at 50 ℃.Add some 3-bromo-1,1 again, (0.06mL is 0.58mmol) and with this mixture heating up to 60 ℃ for the 1-trifluoroacetone.At 60 ℃ after following 24 hours, make this mixture be cooled to envrionment temperature.Add entry (4mL), add EtOAc (4mL) then.Carry out that layer separates and water layer is extracted with EtOAc.With the dry (Na of the organic layer that is merged ₂SO ₄), filter and it is concentrated.With this dissolving crude product in IPA (4mL).In this IPA solution of 1mL, add methylsulfonic acid (0.020mL).With one night of this mixture heating up to 80 ℃ heating.Then, this mixture is cooled to envrionment temperature and it is concentrated, thereby obtain title compound: APCI MS [M+H] ⁺=519.

Embodiment 74h

NaOMe (0.23mL, 1mmol, the MeOH solution of 25 weight %) is joined embodiment 74f, and (409mg is 1mmol) in the mixture in 1-PrOH (2mL).With this mixture heating up to 50 ℃ (reaction-blocking-up temperature).After heating 1 hour, HPLC analysis revealed parent material transforms fully.With this mixture heating up to 70 ℃ and to wherein adding NH ₄OAc (231mg, 3mmol).At 70 ℃ after following 1 hour, with this mixture heating up to 85 ℃.Simultaneously, whenever divided 4 parts to add 3-bromo-1,1 with the amount of every part of 0.033mL at a distance from 30 minutes, and the 1-trifluoroacetone (0.13mL, 1.2mmol).This mixture was heated about 20 hours down at 85 ℃.This mixture is cooled to envrionment temperature and to wherein adding entry (2mL).After stirred for several hour, it is washed (2 * 3mL) through solid collected by filtration and with 1: 1 1-PrOH/ water.This solid is descended dry about 16 hours at 50 ℃ in vacuum drying oven, thereby obtain 0.11g (21%) title compound.

Embodiment 75

(200mg 1mmol) joins 4-(2-(5-(trifluoromethyl)-1H-imidazoles-2-yl) pyridin-4-yl oxygen base)-N1-methylbenzene-1, and (350mg is 1mmol) in the mixture in the 3mL acetonitrile for the 2-diamines with 4-trifluoromethyl lsothiocyanates.After it was stirred 20 minutes at ambient temperature, it transformed the HPLC analysis revealed fully.(553mg is 1mmol) at POCl with thiocarbamide ₃Mixture (3mL) stirs at ambient temperature.After 4 hours, with extremely about 50 ℃ of this mixture heating up.After heating 2 hours, it reacts completely the HPLC analysis revealed.

Embodiment 76

The Raf/Mek filtration test

Damping fluid

Test damping fluid: 50mM Tris, pH7.5,15mM MgCl ₂, 0.1mM EDTA, 1mMDTT

Lavation buffer solution: 25mM Hepes, pH7.4,50mM trisodium phosphate, 500mM NaCl

Stop reagent: 30mM EDTA

Material

Raf, activated: Upstate Biotech #14-352

Mek, non-activity: Upstate Biotech #14-205

³³P-ATP：NEN?Perkin?Elmer?#NEG?602h

96 hole test boards: Falcon U type-end polypropylene board #35-1190

Filtration unit: micropore #MAVM 096 OR

96 hole screen plates: micropore Immobilon 1 #MAIP NOB

Scintillation solution: Wallac OptiPhase " SuperMix " #1200-439

Test conditions

The about 120pM of Raf

The about 60nM of Mek

³³P-ATP?100nM

Reaction times is at room temperature 45-60 minute

Testing program

Raf and Mek are merged to test damping fluid (50mM Tris, pH7.5,15mM MgCl with 2 times final concentration ₂.0.1mM EDTA and 1mM DTT) in and its amount with 15 μ L/ holes is distributed in the Vestolen PP 7052 test board (Falcon U type-end Vestolen PP 7052 96 hole test board #35-1190).Measure background level comprising Mek and DMSO and do not comprise in the hole of Raf.

The raf SU11752 test compound that in the hole that comprises Raf/Mek, adds 10 times of 3 μ L with the 100%DMSO dilution.The test damping fluid of using through adding 2.5 times in 12 μ L/ holes dilutes ³³P-ATP begins the reaction of raf kinase activity.After 45-60 minute, stop this reaction through adding 70 μ L termination reagent (30mM EDTA).Screen plate with 70% ethanol wetting 5 minutes in advance, is cleaned it through filtering with lavation buffer solution then.Then, the sample that derives from reacting hole (90 μ L) is transferred in the screen plate.This screen plate is washed 6 times with lavation buffer solution with microporous filter.These plates are dry and add the scintillation solution (Wallac OptiPhase " SuperMix " #1200-439) in 100 μ L/ holes.Then, measure its CPM with Wallac Microbeta 1450 readers.

Embodiment 77

Test 2: biotinylated Raf screening

External Raf screening

Through ATP, MEK substrate being provided and analyzing the phosphate radical part is come activity from the various hypotypes of measure R af serine/threonine kinase to the transfer of MEK residue.Sf9 insect cell through by the rhabdovirus expression vector transfection of personnel selection Raf reorganization carries out the reorganization hypotype that purifying obtains Raf.The MEK of the no kinase activity of recombinating is carried out mark at expression in escherichia coli and behind purifying with vitamin H.For each test, test compound is diluted with DMSO continuously, the vitamin H-MEK (50nM) with itself and Raf (0.50nM) and no kinase activity adds among the ATP (1 μ M) admixed together at reaction buffer then.Subsequently, should react and at room temperature cultivated 2 hours and come termination reaction through adding 0.5M EDTA.The terminated reaction mixture being transferred to the plate (Pierce) of neutradavin-coating goes up and it was cultivated 1 hour.With rabbit anti--p-MEK (Cell Signaling) as primary antibody and with the resisting of europium mark-rabbit as secondary antibodies, offer an explanation fluorescing system (Wallac) with the DELFIA time and measure by the product of phosphorylation.(time resolved) fluorescence to the time explanation on Wallac 1232 DELFIA photofluorometers carries out reading.Come machine to calculate the concentration (IC that each compound 50% suppresses with XL Fit DAS through non-linear regression ₅₀).

With the operation of embodiment 76 or 77, the compound of embodiment 1-64 all shows to have the raf kinase inhibiting activity, its IC ₅₀Be lower than 5 μ M.

Embodiment 78

The inhibition of melanoma tumor growth

With 3 * 10 ⁶Individual A375M human melanoma cell subcutaneous transplantation is in the right flank of big female Nu/Nu mouse of the 10-12 week of the about 24g of body weight.When tumor average volume reaches about 150mm ³When (transplanting back 17 days), according to gross tumor volume mouse is divided into 4 groups at random, every group of 9 mouse also begin with compound of the present invention it to be treated.Only come mouse is carried out administration with matrix or with the compound administered through oral gavage of 10mg/kg, 30mg/kg or 100mg/kg embodiment 25 every day, administration 14 days, and the administration volume all is 0.2mL.Use twice gross tumor volume of digital caliper measurements weekly.Mean tumour volume is as shown in Figure 1.

Embodiment 79

The inhibition of Raf kinase signal in the melanoma cells

As among the embodiment 78, with 3 * 10 ⁶Individual A375M human melanoma cell subcutaneous transplantation is in the right flank of big female Nu/Nu mouse of the 10-12 week of the about 24g of body weight.When tumor average volume reaches about 150mm ³When (transplanting back 17 days), according to gross tumor volume mouse is divided into 4 groups at random, and only perhaps carries out administration with the compound administered through oral gavage of 10mg/kg, 30mg/kg or 100mg/kg embodiment 25 with matrix every day, the administration volume all is 0.2mL.After

administration

4 and 24 hours the time, with mouse euthansia, results tumour and with its quick freezing.

The tumour that is frozen is melted on ice; Weigh; Then it had Roche Complete; Mini does not have in the RIPA damping fluid of proteinase inhibitor mixture tablet (2/25mL damping fluid), 1mM phenyl methanesulfonamide acyl fluorides (PMSF) and 1X Sigma inhibitors of phosphatases mixture II of EDTA with Roche Magna-lyser and carries out homogenize (under 6500rpm, under 4 ℃, 2 cycles of 1 minute).For every 100mg tumor tissues, add 1mL RIPA dissolving damping fluid.This homogenate under the 14K RPM under 4 ℃ in benchtop microcentrifuge centrifugal 20 minutes, is further carried out homogenize (under 4 ℃, 9K RPM, 2 minutes) to it with Qiagen Qiashredders then.Test with Pierce BCA albumen and to measure protein concentration, the quantity with 20 μ g/ holes loads to each sample in the 4-20%Tris-glycocoll SDS-polyacrylamide gel then.Behind PAGE; Albumen is transferred on the Nitrocellulose film; It was at room temperature blocked (the TBST solution of 5% skimmed milk powder) 1 hour, use (in the blocking-up damping fluid) rabbit polyclonal of dilution in 1: 1000 anti-then-phosphoric acid-ERK1/2 antibody (Cell Signalling #9101), rabbit polyclonal is anti--phosphoric acid-MEK antibody (Ceu Signaling#9121), rabbit polyclonal is anti--ERK1/2 antibody (Cell Signalling #9102) or rabbit polyclonal be anti--MEK antibody (Cell Signalling #9122) surveys a night down at 4 ℃.Then, at room temperature these films are washed 5 times (each 5 minutes) with TBST, in all traces, at room temperature cultivated 1 hour with extent of dilution adding HRP-labelled goat-anti--rabbit antibody (in the blocking-up damping fluid) of 1: 5000 and with it.Then, these films with 5 times (each 5 minutes) of TBST washing, are cultivated these films 4 minutes with PierceSuper-Signal, made its exposure then on film, the time shutter is 1 second to 20 minutes.The result of sampling in 4 and 24 hours is respectively shown in Fig. 2 A and 2B after the administration.

Embodiment 80

The inhibition of colorectal carcinoma tumor growth

With 2 * 10 ⁶Individual HT29P human colon cancer cell subcutaneous transplantation is in the right flank of big female Nu/Nu mouse of the 10-12 week of the about 24g of body weight.When tumor average volume reaches about 250mm ³(transplanting back 14 days) is divided into 4 groups according to gross tumor volume with mouse at random, and every group of 10 mouse also begin with compound of the present invention it to be treated.Only with matrix or with the compound of 10mg/kg, 30mg/kg or 100mg/kg embodiment 25 through every day oral gavage come mouse is carried out administration, administration 14 days, the administration volume is 0.2mL.Use twice gross tumor volume of digital caliper measurements weekly.Mean tumour volume as shown in Figure 3.

Embodiment 81

The inhibition of Raf kinase signal in the colon cancer cell

With 3 * 10 ⁶Individual HT29P human colon cancer cell subcutaneous transplantation is in the right flank of big female Nu/Nu mouse of the 10-12 week of the about 24g of body weight.When tumor average volume reaches about 150mm ³(transplanting back 17 days) is divided into four groups and begin with compound of the present invention it to be treated according to gross tumor volume with mouse at random.Only with matrix or with the compound of 10mg/kg, 30mg/kg or 100mg/kg embodiment 25 through every day oral gavage come mouse is carried out administration, administration 5 days, the administration volume is 0.2mL.After

administration

1,4 and 24 hour the time, with mouse euthansia, results tumour and with its quick freezing.Then, according to the operation of embodiment 79 tumour that is frozen is handled.The result of 1,4 and 24 hour sample is respectively shown in Fig. 4 A, 4B and 2C after the administration.

Embodiment 82

The compound of embodiment 1 is to the external Raf test of the inhibition of Raf kinase signal in the external biological chemical test

With following biotinylated test determination the compound of embodiment 1: { 1-methyl-5-[2-(5-three fluoro-methyl isophthalic acid H-imidazoles-2-yls)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine is to the restraining effect of wild-type B-Raf, wild-type c-Raf and mutant B-Raf (V600E).The MEK substrate of the no kinase activity through ATP, reorganization are provided also analyzes the kinase activity of the various hypotypes of measure R af serine/threonine kinase to the transfer of MEK residue to the phosphate radical part.The total length MEK that will have the reorganization of deactivation K97R ATP combining site sudden change (making its no kinase activity) carries out mark with vitamin H to it at expression in escherichia coli and behind purifying.With N-terminal (his) ₆Mark carries out subclone to MEK cDNA, it is also passed through the nickel affinity chromatogram at expression in escherichia coli, then through the MEK substrate of anionresin by purification of Recombinant in the intestinal bacteria lysate.Be concentrated into 11.25 μ M with final MEK substrate preparation biotinylation (Pierce EZ-Link Sulfo-NHS-LC-vitamin H) and with it.Through carrying out B-Raf, c-Raf and the mutant B-Raf that purifying obtains to recombinate by the sf9 insect cell that infects with corresponding human Raf recombinant expression vector.Come the Raf hypotype of reorganization is carried out purifying through interaction of Glu antibody or metals ion chromatography.

For each test, the compound of embodiment 1 is used the DMSO serial dilution, then that itself and B-Raf, c-Raf or mutant B-Raf (each 0.50nM) is admixed together.Vitamin H-MEK the substrate (50nM) that in the reaction buffer that is added with ATP (1 μ M), adds no kinase activity.This reaction buffer comprises 30mM Tris-HCl, pH7.5,10mM MgCl ₂, 2mM DTT, 4mMEDTA, 25mM β-glycerophosphate, 5mM MnCl ₂And 0.01%BSA/PBS.Subsequently, should react and at room temperature cultivated 2 hours and through adding 0.5M EDTA termination reaction.The reaction mixture that is terminated transferred in the plate (Pierce) that neutradavin-applies and with it cultivated 1 hour.With rabbit anti--p-MEK (Cell Signaling) as primary antibody and with the resisting of europium mark-rabbit as secondary antibodies, offer an explanation fluorescing system (Wallac) with the DELFIA time and measure by the product of phosphorylation.The fluorescence of time for reading explanation on Wallac 1232 DELFIA photofluorometers.With XL Fit DAS, calculate embodiment 1 compound concentrations (IC for 50% inhibition through non-linear regression ₅₀).

The result:

Shown in following table 3, the compound of embodiment 1 shows effective restraining effect (IC to B-Raf, c-Raf and mutant B-Raf (V600E) activity ₅₀＜0.1 μ M).

The compound of table 3: embodiment 1 is to the active vitro efficacy of Raf

Shown in top table 3, the compound of embodiment 1 shows effective inhibition activity to wild-type B-Raf hypotype, wild-type c-Raf hypotype and mutant B-Raf (V600E) Raf kinases.As shown in Figure 5, think that the Raf kinases is the elementary Ras effector in MAPK (Ras/Raf/MEK/ERK) signal transduction pathway.The Raf kinases is by Ras activation and phosphorylation and activation Mek1 and Mek2, its next in the MAPK approach again activation mitogen activatory kinases 1 and 2 (MAPK).Known Raf kinases influence and adjusting cell proliferation, differentiation, survival, carinogenicity transform and apoptosis.Shown that the B-Raf hypotype is to participate in the most activated form of Raf of signal transduction and very crucial in the propagation of Ras signal.

Shown in following table 4, in many human cancers, all relate to the MAPK signal transduction pathway.Whole human cancers 15% in all found Ras sudden change (activatory).Whole human cancers 30% in all found ERK sudden change (overactivation).With the sudden change of the oncogene of related to cancer in some members of this approach, be common, for example mutant B-Raf (V600E) betide in about 70% the melanoma with about 12% colorectal carcinoma in (people such as Davies, the same; People such as Yuen, people such as the same and Brose, the same).

Mutant signaling molecule in the table 4:MAPK approach and the relation between the clinical effectiveness difference

See Sebolt-Leopole and Herrera, Nature Reviews Cancer (4): 937 (2004).

Shown in top table 4; The mutant of activated B-Raf (V600E) is the important target spot of cancer therapy, and this is because its expression is the indicator of poor prognosis, and it is that composition is active; And driven some tumours, comprised melanoma, papillary thyroid carcinoma, ovarian cancer and colorectal carcinoma.The kinase whose suppressor factor of wild-type Raf of verified before also mutation inhibiting type B-Raf is promising therapeutical agent in cancer therapy.For example, shown that siRNA has damaged ERK signal and propagation (Dibb, people such as NJ., the Nature Reviews Cancer (4): 718,2004) in the melanoma cell series to the consumption of mutant B-Raf.Therefore; Compound that it should be noted that embodiment 1 is higher to inhibition even the comparison wild-type B-Raf of mutant B-Raf, thereby has proved that this compound suppresses the effectiveness of Raf in the treatment of the disease (comprising melanoma, ovarian cancer, papillary thyroid carcinoma and colorectal carcinoma) of Raf-mediation.

Embodiment 83

The compound of embodiment 1 is to mutant B-Raf kinase signal in the test that with the cell is the basis

Inhibition

1.ERK the restraining effect of phosphorylation

Method:

A kind of with the cell be the basis test in, with two kinds of melanoma cell series---A375M (mutant B-Raf V600E) and SKMEL-28 (mutant B-Raf V600E) measure the restraining effect of embodiment 1 compound.In SKMEL-28 cell and A375M cell; With embodiment 1 compound: after the serial dilution thing of { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine was handled, phosphorylation was analyzed to ERK.Measure EC through fitting data in four parametric lines ₅₀Value.

The result:

Shown in following table 5, reducing through phosphoric acid-ERK when measuring, the compound of embodiment 1 has suppressed the kinase whose activity of mutant B-Raf (V600E) in SKMEL-28 cell and the A375M cell.

The restraining effect of table 5: embodiment 1 compound in the melanoma cell series of expressing mutant Raf-B

2.MEK the restraining effect of phosphorylation

Method:

A kind of with the cell be the basis test in; With three kinds of melanoma cell series---A375M (mutant B-Raf V600E), SKMEL-2 (wild-type Raf; Mutant N-Ras) and CHL-1 (wild-type Raf, wild-type Ras) measure the restraining effect of embodiment 1 compound.Three kinds of clones are cultivated containing under 37 ℃ in 0.1% foetal calf serum of compound of embodiment 1 that concentration is 0.1 μ M, 0.5 μ M, 1 μ M, 5 μ M and 10 μ M.After cultivating 4 hours, with western blot analysis to MEK phosphorylation analyze.

The result:

Shown in result such as Fig. 6 A, 6B and the 6C.Shown in Fig. 6 A, the compound of embodiment 1 is Raf downstream signal effective inhibitors in A375M cell (Fig. 6 A), SKMEL-2 cell (Fig. 6 B) and the CHL-1 cell (Fig. 6 C) in the concentration range of 0.1 μ M to 10 μ M.

3. the restraining effect of non-set dependent cell growth

Translated into antiproliferative activity for the inhibition that proves Raf, be used in making an experiment of growing in the soft agar like various clones listed in the following table 6 and people's tumour isolate compound to embodiment 1.

Soft agar proliferation test: for each listed in the following table 6 clone, its amount with 500 cells/100 μ L is inoculated in the Corning 96 holes flat Ultra Low Attachment Micro plates (Corning #3474).In perfect medium, add 1%Seakem GTG agarose (50 μ L/ hole), make its curing, in each hole, add 100 μ L perfect mediums then.The compound that in the substratum that does not contain serum, prepares embodiment 1 with the final concentration of 5%DMSO: the serial dilution thing of { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine, and in each hole, add the diluted compound (the DMSO final concentration is 0.5%) of 25 μ L.In this test, also comprise the control wells that comprises 0.5%DMSO that does not have compound.After cell is cultivated 7 days with this compound, in each hole, add 25 μ L Alamar Blue (Trek Diagnostic Systems #00-100) and it was cultivated 4 hours down at 37 ℃.With the fluorescent plate reader this plate is carried out reading, excitation wavelength is 530nm, and emission wavelength is 590nm.Measure EC through fitting data in a kind of four parametric lines ₅₀Value.

The lineup's tumour isolate that also is used in growth in the soft agar (Oncotest, GmbH, Freiburg, Germany) is tested the compound of embodiment 1.Tumour is separated in patient's body, its form with tumor fragment is analyzed it at immunocompromised mouse interior generation and with aforesaid method.

The result: shown in following table 6, the compound of embodiment 1 all has effective antiproliferative effect to clone and people's tumour isolate of expressing mutant B-Raf, mutant K-Ras and mutant N-Ras.

Table 6: soft agar proliferation test: with the inhibition of embodiment 1 compound

The compound of embodiment 1 has proved this compound effective antiproliferative activity in the tumor cell line of expressing mutant B-Raf to the inhibition activity of the many clones shown in the top table 6 and people's tumour isolate.This compound shows effective restraining effect to mutant B-Raf melanoma cells in the scope of＜0.0098 to 0.07 μ M.This compound shows similar inhibition degree to mutant B-Raf colorectal cell system in the scope of 0.026 μ M to 0.13 μ M.Prove that also this compound has effective antiproliferative activity in two kinds of colorectal cancer cells of the expression mutant K-Ras that is tested (0.07 μ M to 0.016 μ M), confirm that the inhibition of B-Raf/c-Raf in the background of upper reaches K-Ras sudden change has produced antiproliferative activity.

Consistent with the result who derives from above-mentioned clone, the compound of embodiment 1 has proved that to the effect of people's tumour isolate it is to mutant B-Raf melanoma (EC ₅₀=0.055 μ M and 0.20 μ M) have the strongest restraining effect, secondly be to N-Ras mutant melanoma (EC ₅₀=0.57 μ M) restraining effect.The EC that a kind of pancreas tumor and a kind of colorectum tumour have 1 μ M scope ₅₀Remaining tumour has provided the EC that is higher than 1 μ M ₅₀Value.Because tumour is separated in patient's body and made it at immunocompromised mouse interior generation with the form of tumor fragment, think that therefore people's tumour of separating in patient's body represented a kind of than clone disease model more accurately.Therefore, do not select to make it on plastics, to grow and it has still kept some tumour system structures of former.

What is interesting is, notice that the compound of embodiment 1 has the inhibition activity that scope is higher than 1 μ M in renal cell carcinoma tumour isolate.Though in these specific tumours, do not record this genotype, known renal cell carcinoma tumour is not expressed mutant Ras or mutant B-Raf usually.Therefore, the compound of embodiment 1 seems signaling molecule, particularly Raf and the Ras kinase molecule of specific inhibition MAPK approach.

Embodiment 84

The processing of carrying out with the compound of embodiment 1 has caused A375M (B-Raf V600E) human melanoma different

Plant the tumor regression in the graft model

Method: with 3 * 10 ⁶Individual A375M human melanoma cell subcutaneous transplantation is in the right flank of big female Nu/Nu mouse of the 10-12 week of the about 24g of body weight.When tumor average volume reaches about 150mm ³When (transplanting back 17 days), according to gross tumor volume mouse is divided into 4 groups at random, every group of 9 mouse also begin to treat with the compound of embodiment 1.Only with matrix or with the compound of 10mg/kg, 30mg/kg or 100mg/kg embodiment 1 through every day oral gavage come mouse is carried out administration, administration 14 days, the administration volume is 0.1mL.In 100%PEG to the compound of embodiment 1: { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine is prepared.Use twice gross tumor volume of digital caliper measurements weekly.

Western blot analysis

After the 14th administration 8 and 24 hours, with mouse euthansia, results tumour and with its quick freezing.The tumour that is frozen is melted on ice; Weigh; Then it had Roche Complete; Mini does not have in the RIPA damping fluid of proteinase inhibitor mixture tablet (2/25mL damping fluid), 1mM phenyl methanesulfonamide acyl fluorides (PMSF) and 1X Sigma inhibitors of phosphatases mixture II of EDTA with Roche Magna-lyser and carries out homogenize (under 6500rpm, under 4 ℃, 2 cycles of 1 minute).For every 100mg tumor tissues, add 1mL RIPA dissolving damping fluid.This homogenate under the 14K RPM under 4 ℃ in benchtop microcentrifuge centrifugal 20 minutes, is further carried out homogenize (under 4 ℃, 9K RPM, 2 minutes) to it with Qiagen Qiashredders then.Test with Pierce BCA albumen and to measure protein concentration, the quantity with 20 μ g/ holes loads to each sample in the 4-20%Tris-glycocoll SDS-polyacrylamide gel then.Behind PAGE; Albumen is transferred on the Nitrocellulose film; It was at room temperature blocked (the TBST solution of 5% skimmed milk powder) 1 hour, use (in the blocking-up damping fluid) rabbit polyclonal of dilution in 1: 1000 anti-then-phosphoric acid-ERK1/2 antibody (Cell Signalling #9101), rabbit polyclonal is anti--phosphoric acid-MEK antibody (Cell Signaling#9121), rabbit polyclonal is anti--ERK1/2 antibody (Cell Signalling #9102) or rabbit polyclonal be anti--MEK antibody (Cell Signalling #9122) surveys a night down at 4 ℃.Anti--Bim antibody (Chemicon with dilution in 1: 1000; #AB 17003), anti--cyclin D1 antibody cloning 5D4 (Upstate; #05-263), anti--p27Kip-1 (182-198) antibody (Calbiochem; #506127), anti--phosphoric acid-AKT (S473) antibody (Cell Signaling, #9271), anti--phosphoric acid-Akt (T308) antibody (Cell Signaling #9275) is with anti--phosphoric acid-always Akt antibody (Cell Signaling #9272) is analyzed the adjusting of downstream affinity tag.

Then, at room temperature these films are washed 5 times (each 5 minutes) with TBST, in all traces, at room temperature cultivated 1 hour with extent of dilution adding HRP-labelled goat-anti--rabbit antibody (being arranged in the blocking-up damping fluid) of 1: 5000 and with it.Then, these films with 5 times (each 5 minutes) of TBST washing, are cultivated these films 4 minutes with Pierce Super-Signal, made its exposure then on film, the time shutter is 1 second to 20 minutes.

The result:

Fig. 7 A representes when handling with the oral dosage of 10mg/kg, 30mg/kg or 100mg/kg embodiment 1 compound the figure of the dose response that A375M (B-Raf V600E) human melanoma gross tumor volume on average reduces in the mouse body.Shown in Fig. 7 A, the compound of embodiment 1 has the effective antitumour activity with oral dosage dependency mode.Under the oral dosage of this compound 100mg/kg, in whole 9 mouse of being tested, all observed tumor regression.

In Fig. 7 B and Fig. 7 C, represented respectively with the dosage of 10mg/kg, 30mg/kg and 100mg/kg with the 14th administration of compound of embodiment 1 after the result of 8 hours and 24 hours western blot analysises.These Western blot data show that this compound has suppressed MEK phosphorylation (it has induced tumor regression) under 100mg/kg dosage and shown in Fig. 7 C, after the administration, this MEK restraining effect continues more than 24 hours the last time.

Shown in Fig. 7 D; The analysis revealed BIM (apoptotic mark) and the p27Kip (mark of cell cycle arrest) that after the 14th administration, tumour lysate downstream biomarker were regulated in 24 hours increase, and cyclin D (cell cycle inhibition) reduces.These results have confirmed that the compound of embodiment 1 has suppressed the Raf signal in the MAPK approach.

Embodiment 85

The treatment of carrying out with the compound of embodiment 1 has suppressed the melanoma tumor growth

The compound of the embodiment 1 inhibition activity in melanoma tumor model MEXF276 (mutant B-RafV600E) and melanoma tumor model MEXF 1341 wild-type B-Raf, mutant N-Ras (Q61K) is tested.

Method: with human melanoma MEXF276 (mutant B-Raf V600E) the tumour cell subcutaneous transplantation of continuous passage in the back flank of big female Nu/Nu mouse of 10-12 week.When tumor average volume reaches about 65mm ³The time, according to gross tumor volume these mouse random packet and compound { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine of beginning with embodiment 1 are treated.Because known MEXF276 model is cachectic to a certain extent, expection has some toxicity in the control mice of treating, for body weight serious in the prophylactic agent treatment group reduces, and following such intermittently dosage regimen of using.Only mouse is carried out administration: at the 0th, 2,4,6,14,16 and 20 day 10mg/kg with matrix or the dosage regimen administered through oral gavage below embodiment 1 compound; At the 0th, 2,14,16 and 20 day 30mg/kg; With at the 0th, 2,14,16 and 20 day 100mg/kg.

For the MEXF1341 model, with the human melanoma MEXF1341 tumour cell subcutaneous transplantation of continuous passage in the back flank of female Nu/Nu mouse.When tumor average volume reaches about 78mm ³The time, according to gross tumor volume with the mouse random packet and begin to treat with the compound of embodiment 1.Only mouse is carried out administration: at the 0th, 2,4,6,10,12,18 and 20 day 10mg/kg with matrix or the dosage regimen administered through oral gavage below embodiment 1 compound; At the 0th, 2,4,6,10,12,18 and 20 day 30mg/kg; With at 0,2,4,6,10,12 and 20 day 100mg/kg.

The last time after the administration 4 hours the time, with the mouse euthansia that derives from MEXF276 and MEXF1341 model, results tumour and with its quick freezing.Subsequently, as described in the top embodiment 84, analyze with the adjusting that Western blot is regulated (pMEK) and downstream mark (BIM, p27Kip and pAKT) to the target of the lysate that derives from these tumours.

The result:

Fig. 8 A representes when treating with the compound of embodiment 1, the figure of the average reduction of MEXF276 (B-Raf V600E) melanoma cancerous tumour volume in the mouse body.Result shown in Fig. 8 A shows that the compound of embodiment 1 shows significant tumor growth and suppresses under 10mg/kg in the MEXF276 isolate, and is greater than or equal to 50% tumor regression in whole 8 mouse, all showing under 30mg/kg and the 100mg/kg.Analysis (Fig. 8 C) to pMEK phosphorylation in the tumour lysate (Fig. 8 B) and downstream and the adjusting of downstream biomarker proves that kind shown in the reduction of phosphoric acid-MEK among Fig. 8 B, the mutant B-Raf activity inhibited in the MEXF276 tumour.Shown in Fig. 8 C, observe BIM (apoptotic mark) and p27Kip (cell cycle arrest) increase and phosphoric acid-AKT (survival approach signal) and reduce, confirm to have suppressed the Raf kinase activity in the MAPK approach.

Fig. 9 A be expression when treat with the compound of embodiment 1, the figure of the average inhibition that MEXF1341 in the mouse body (N-Ras Q61K) melanoma cancerous tumour is grown.Result shown in Fig. 9 A shows that the compound of embodiment 1 has caused significant tumor growth to suppress (high to 70% inhibition) under the dosage of 30mg/kg and 100mg/kg in MEXF 1341 mutant N-Ras (N-Ras Q61K) melanoma tumor model, but do not have induced tumor to disappear.Shown in Fig. 9 B, opposite treat back 20 days analysis revealeds with the said compound of 100mg/kg with the result who in MEXF276 (mutant B-Raf) model, obtains to phosphoric acid-MEK, do not observe phosphoric acid-MEK and reduce.In addition, though there are some signs to show that it has influenced some signaling molecules in the MAPK approach Raf downstream in MEXF 1341 models, its effect significantly is lower than observed effect in the MEXF276 model.For example, shown in Fig. 9 C, p27Kip level (cell cycle arrest) increases in 30mg/kg and 100mg/kg group, and it shows growth-inhibiting, and observes the slight increase of apoptosis mark BIM.Therefore; As if the compound of embodiment 1 has effective activity in MEXF276 (mutant B-Raf) xenograft models; Caused tumor regression; Activity (having caused the tumor growth inhibition) significant that but effectiveness is lower is also arranged in MEXF1341 (wild-type B-Raf, mutant N-Ras) xenograft models.

Embodiment 86

The treatment of carrying out with the compound of embodiment 1 has suppressed the human colorectal cancer tumor growth

The inhibition activity of compound in colorectal carcinoma xenograft models HCT-116 (mutant K-Ras G13D), HT-29 (B-Raf V600E) and acute leukemia xenograft models MV4-11 (FLT3 ITD) to embodiment 1 tested.

Method: with 5 * 10 ⁶Individual HCT-116 (mutant K-Ras G13D) human colorectal cancer cell subcutaneous transplantation is in the back flank of big female Nu/Nu mouse of the 10-12 week of the about 24g of body weight.When tumor average volume reaches about 212mm ³The time, only mouse is carried out administration with matrix or dosage regimen administered through oral gavage below embodiment 1 compound: the 1st day with every at a distance from 2 days (q2d) with the oral tube feed of 10mg/kg, 30mg/kg and 100mg/kg, 28 days altogether.The 4th hour, 8 hours and 24 hours are with satellite (Satellite) mouse euthansia and gather in the crops tumour after the 3rd administration.Subsequently, as described in the top embodiment 84, with Western blot the target of the lysate that derives from these tumours is regulated (pMEK) and analyze.

Following such to second kind of human colorectal cancer model---HT-29 (B-Raf V600E) makes an experiment.With 2 * 10 ⁶Individual HT-29 cell subcutaneous transplantation is in the back flank of big female Nu/Nu mouse of the 10-12 week of the about 24g of body weight.When tumor average volume reaches about 167mm ³The time, only mouse is carried out administration with matrix or dosage regimen administered through oral gavage below embodiment 1 compound: the 1st day with every at a distance from 2 days (q2d) with the oral tube feed of 10mg/kg, 30mg/kg and 100mg/kg, 28 days altogether.

Following such to people's acute monocytic leukemia xenograft models---MV4-11 (FLT3 ITD) makes an experiment: with 5 * 10 ⁶Individual MV4-11 cell subcutaneous transplantation is in the back flank of big female Nu/Nu mouse of the 10-12 week of the about 24g of body weight.When tumor average volume reaches about 190mm ³The time, only mouse is carried out administration with matrix or dosage regimen administered through oral gavage below embodiment 1 compound: the 1st day with every at a distance from 2 days (q2d) with the oral tube feed of 10mg/kg, 30mg/kg and 100mg/kg, 16 days altogether.After the 3rd administration 4 hours with satellite mouse euthansia and gather in the crops tumour.Subsequently, as described in the top embodiment 84, with Western blot the target of the lysate that derives from these tumours is regulated (pMEK) and analyze.

The result:

The result who in Figure 10 A-D, has represented HCT-116 research.Figure 10 A representes when treating with the compound of 100mg/kg embodiment 1, the figure of the average reduction of HCT-116 (K-Ras G13D) colorectal carcinoma gross tumor volume in the mouse body.Shown in Figure 10 B-10D, phosphoric acid-MEK analysis revealed that 4 hours (Figure 10 B), 8 hours (Figure 10 C) and 24 hours (Figure 10 D) carry out after the 3rd administration has been observed the reduction of phosphoric acid-MEK.

Figure 11 representes when treating with the compound of embodiment 1 figure of the average reduction of HT-29 (B-Raf V600E) colorectal carcinoma gross tumor volume.As shown in Figure 11, under 30mg/kg and 100mg/kg, observe tumor regression.

The result who in Figure 12 A-B, has represented MV4-11 research.Figure 12 A representes when treating with the compound of embodiment 1 figure of the average inhibition that MV4-11 acute monocytic leukemia tumor is grown in mouse.The MV4-11 tumour cell is by mutant receptor tyrosine kinase (MV4; 11, FLT3 ITD) drive.As illustrated, the compound of embodiment 1 has caused significant tumor growth to suppress in the MV-11 model, still, has not both had the sign (Figure 12 A) of tumor regression, does not also observe the inhibition (Figure 12 B) of MEK signal.Though do not hope to receive the constraint of any theory, as described in the embodiment 87-88, in the MV4-11 model, the effect that tumor growth suppresses possibly be this compound anti--result of angiogenic activity, it mainly is the inhibition through VEGFR-2.The general introduction of the data that obtained by the assessment of in above-mentioned melanoma, colorectal carcinoma and white blood disease xenograft models, the effect of the compound of embodiment 1 being carried out is provided in the table 7 below.

The active general introduction of the compound of table 7: embodiment 1 in various xenograft models

Generalized data from table 7; Shown in Fig. 6-12; And described in embodiment 82-86; The compound of embodiment 1 all is effectively in each xenograft models that the wherein B-Raf that is tested has taken place to suddenly change, and in all three kinds of models (A375M, MEXF276 and HT29) of being tested, has all caused tumor regression and target to regulate.

Embodiment 87:

The Tyrosylprotein kinase inhibition test

1. biochemical test:

Through ATP and the peptide that suits being provided or comprising the albumen of the tyrosine amino-acid residue that is used for phosphorylation and the phosphate radical part is analyzed the kinase activity of measuring many protein tyrosine kinases to the transfer of tyrosine residues.The recombinant protein that is equivalent to the cytoplasmic structure territory of VEGFR2, PDGFR β, CSF-1R and c-Kit is to obtain through carrying out purifying by the Sf9 insect cell with corresponding human VEGFR-3 2, PDGFR β, CSF-1R and the transfection of c-Kit recombination rhabdovirus expression vector.For each test; With compound { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl-phenyl)-amine serial dilution in DMSO of embodiment 1, it is admixed together then itself and suitable kinase reaction damping fluid to be added ATP (concentration of ATP equals separately Km value perhaps only a little less than described Km value).Add said kinase protein and suitable biotinylated peptide substrates, thereby obtain the final volume of 50-100 μ L.These reactions were at room temperature cultivated 1-3 hour, stop this reaction through adding 25-50 μ L 45mM EDTA, 50mM Hepes pH7.5 then.The reaction mixture (75 μ L) that has been terminated is transferred in the microtiter plate (Boehringer Mannheim) of streptavidin-coating and and cultivated 1 hour it.(the DELFIA test damping fluid that is used for antibody dilution has been added 1mMMgCl under the situation of having carried out some modifications ₂), with anti--phosphotyrosine antibody PT66 of europium mark, offer an explanation the peptide prod that fluorescing system (Wallac or PE Biosciences) is measured phosphorylation with the DELFIA time.The fluorescence of time for reading explanation on Wallac 1232DELFIA photofluorometer or PE Victor II multiple signal reader.Use XL Fit DAS, calculate each compound concentrations (IC for 50% suppresses with non-linear regression ₅₀).

At 50mM Hepes pH7.0,2mM MgCl ₂, 10mM MnCl ₂, VEGFR2 kinases (0.05 μ g/mL) is made an experiment in 1mM NaF, 1mM WR 34678 (DTT), 1mg/mL bovine serum albumin (BSA), 1 to 30 μ M ATP and the biotinylated peptide substrates of 0.25 μ M " GGGGQDGKDYIVLPI " (SEQ ID NO:1).

For the PDGFR kinase assay; Use has 120 μ g/mL enzymes of the buffer conditions identical with above-mentioned buffer conditions; Just ATP and peptide substrates concentration are become 1.4 μ M ATP and use the biotinylated peptide substrates of 0.25 μ M " GGGGQDGKDYIVLPI " (SEQ IDNO:1).

At test damping fluid (50mM HEPES pH7.0,5mM MgCl ₂, 10mM MnCl ₂, 0.1%BSA, 1mM DTT, 0.01% tween, final pH is 7.5), the kinase activity to CSF-1R in 1 μ M ATP and the biotinylated peptide substrates of 50nM " EEEEAYGWLNF " (SEQ ID NO:2) makes an experiment.

Through being provided, ATP and the recombinant protein (deriving from Proquinase) that is equivalent to the cytoplasmic structure territory of c-Kit acceptor measure the kinase activity of c-Kit.C-Kit kinase protein (2nM) and biotinylated peptide substrates (1 μ M) " GGLFDDPSWNVQNL " (SEQ ID NO:3) are joined reaction buffer add among the ATP (8 μ M), thereby obtain the final volume of 100 μ L.The reaction buffer that is used for c-Kit is 50mM HEPES pH7.5,1mM NaF, 2mM MgCl ₂, 10mM MnCl ₂With 1mg/mL BSA.Should react and at room temperature cultivate 2 hours also through adding 50 μ L 45mMEDTA, 50mM HEPES, pH7.5 stops this reaction.The reaction mixture (75 μ L) that has been terminated is transferred in the microtiter plate (BoehringerMannheim) of streptavidin-coating and and cultivated 1 hour it.(the DELFIA test damping fluid that is used for the antibody detection has been added 1mM MgCl under the situation of having carried out some modifications ₂), with anti--phosphotyrosine antibody PT66 of europium mark, offer an explanation the peptide prod that fluorescing system (Wallac or PE Biosciences) is measured phosphorylation with the DELFIA time.The fluorescence of time for reading explanation on Wallac 1232 DELFIA photofluorometers or PE Victor II multiple signal reader.Use XL Fit DAS, calculate the compound concentrations (IC of embodiment 1 for 50% suppresses with non-linear regression ₅₀).

The result: shown in following table 8, the compound of embodiment 1 is effective VEGFR-2, c-Kit, PDGFR-β and CSF-1R suppressor factor.

The compound of table 8: embodiment 1 is to the inhibition of Tyrosylprotein kinase

Target	The compound biological chemistry IC of embodiment 1 ₅₀	The compound of embodiment 1 is the EC on basis with the cell ₅₀
			VEGFR-2	?0.07μM	0.03μM
c-Kit	?0.02μM	1.1μM
			PDGFR-β	?0.0032μM	0.7μM
CSF-1R	?0.20μM	ND

Following such, also in order to the activity of the cell target molecule shown in the compound his-and-hers watches 8 of embodiment 1 that be basic experimental measurement.

After handling with the compound of embodiment 1, as the phosphoric acid that measures with Western blot-VEGFR reduce measured (not shown), the target of HMVEC cell is regulated and is shown its EC with 0.03 μ M ₅₀Suppressed the VEGFR-2 phosphorylation of VEGF mediation.

As the phosphoric acid that measures with ELISA-c-Kit reduce measured, after treating with the compound of embodiment 1, its EC of analysis revealed that the inhibition of c-Kit in the Mo7e cell is carried out with 1.1 μ M ₅₀Suppressed the c-Kit phosphorylation.

As the phosphoric acid that measures with ELISA-PDGFR-β reduce measured, after treating with the compound of embodiment 1, its EC of analysis revealed that the inhibition of PDGFR-β in the MG63 cell is carried out with 0.7 μ M ₅₀Suppressed phosphoric acid-PDGFR-β.

Embodiment 88

The inhibition that blood vessel takes place

In order further the compound of embodiment 1 to be studied the effect of VEGFR-2, this compound is assessed with CHO-VEGF Matrigel blood vessel generation model.

Method: before beginning one's study, make 110Nu/Nu mouse (n=10/ group) adapt to a week.At the 1st day, will be arranged in 5 * 10 of 0.5mL Matrigel ⁶Individual VEGF-CHO cell is subcutaneously injected into the epigastrium of mouse.At the 1st day,, give the compound of mouse oral matrix, 10mg/kg, 30mg/kg or 100mg/kg embodiment 1 according to the dosage regimen of qdx5.After 5 days, take out this Matrigel plug in the mouse body and wherein HC is carried out quantitatively.

The result:

Figure 13 is illustrated in the compound of 10mg/kg, 30mg/kg and 100mg/kg embodiment 1 to treat the inhibiting figure of back to the blood vessel generation of VEGF-mediation in the CHO-VEGF Matrigel model.As shown in Figure 13, this compound administration was significantly suppressed the blood vessel generation of VEGF-mediation in 5 days.

Embodiment 89

The influence of dosage regimen

For the relation between the compound concentration in mutant B-Raf inhibition, tumour response and the blood plasma is assessed, in A375M human melanoma xenograft models, carry out the dosage regimen research of embodiment 1 compound.

Shown in Fig. 7 A, in the A375M model, set up clearly dosage-response relation of embodiment 1 compound.Data among Fig. 7 A show, when the every day administration, the compound of embodiment 1 has been induced tumor regression under 100mg/kg, and tumor regression relevant with the lasting inhibition of mutant B-Raf (shown in the reduction of phosphoric acid-MEK among Fig. 7 B).But in this dosage regimen, the compound of embodiment 1 can not be by well tolerable under the dosage level of 30mg/kg and 100mg/kg, and this is that the initial weight average of mouse has lost 10% because during by the 14th day.Therefore, be described below and such further the most effective dosage of 100mg/kg assessed.

Method:

The same with embodiment 84, with 3 * 10 ⁶Individual A375M human melanoma cell subcutaneous transplantation is in the right flank of big female Nu/Nu mouse of the 10-12 week of the about 24g of body weight.When tumor average volume reaches about 200mm ³The time, according to gross tumor volume mouse being divided into 4 groups at random, every group of 9 mouse also begin to treat with the compound of embodiment 1.Only come mouse is carried out administration administration 32 days: q2d, q3d or q4d, 100mg/kg, administration 28 days with the compound administered through oral gavage of embodiment 1 with matrix or with following dosage regimen.

In this research, the satellite group of tumor-bearing mice is carried out administration regulate with the target in the monitoring tumour.In q2d group administration 5 times with after q4d group administration 3 times, in the mouse body, collect tumour and blood plasma at each time point.As described in the embodiment 84, tumour is handled so that phosphoric acid-MEK level is carried out western blot analysis, and separated plasma is to measure levels of drugs.

The result:

Figure 14 A representes when treating with the compound of 100mg/kg embodiment 1 with the dosage regimen of q2d, q3d or q4d the figure of the average reduction of mouse A375M melanoma gross tumor volume.Shown in Figure 14 A, the compound of embodiment 1 has produced obvious effect with the scheme of q2d, q3d or q4d with the 100mg/kg oral administration.Western blot analysis shown in Figure 14 B is illustrated in the q2d sample, and after administration 48 hours, for the contrast of matrix treatments, the intravital tumour of mouse of treating with said compound had the phosphoric acid-MEK level of reduction.In the q4d sample, during to 72 hours, in three tumours only a tumour have the phosphoric acid-MEK level of reduction, during to 96 hours, all tumours of handling with compound all have the phosphoric acid-MEK level suitable with the tumour of matrix treatments.Shown in Fig. 7 B, these results are consistent with the result who obtains according to the q2d dosage regimen.

Shown in following table 9, when weighing through the body weight reduction, the dosage regimen of q3d and q4d can be tolerated by test mice better.

Table 9: the A375M heterograft administration with 100mg/mL embodiment 1 compound carries out is studied

In a word, when taking all factors into consideration target adjusting data and efficacy data, the dosage regimen of q2d or q3d has produced the most effectively tumor regression under the situation with maximum main body tolerance.

Embodiment 90

The research of target plasma concns

Described in top embodiment 89, in the mouse body of treating, get serum sample with the compound of embodiment 1.Measure the drug level in these serum samples and come the result is represented with the form of the compound concentration-time diagram among Figure 15.Can regulate data through considering the target shown in Figure 14 A and Figure 14 B, estimate the medicine threshold concentration that target is regulated by Figure 15.Shown in Figure 14 B, to all time points of 48 hours, for the tumour of matrix treatments, phosphoric acid in the tumour of compound treatment-MEK level reduced after administration, and therefore, corresponding drug level necessarily is higher than this threshold value.Shown in Figure 14 C, after administration 72 and 96 hours, there is not target to regulate, therefore corresponding drug level necessarily is lower than this threshold value.In a word, it is about 50,000 to 80 that the threshold value of estimating said compound should be, 000ng/mL, and as about 70,000ng/mL (135 μ M).

What is interesting is, notice the EC that the target plasma concns in the above-mentioned mouse heterograft research is regulated than the middle target of external A375M cell (.16 μ M) ₅₀(seeing table 5) high about 1000 times.This species diversity can combine to make an explanation to a great extent with plasma proteins, and this is because the compound of embodiment 1 is protein bound in blood plasma more than 99.9%.Consider this point, guestimate free drug concentration is about 0.135 μ M, and it approaches the external EC of the 0.16 μ M that the target adjusting records in the A375M cell ₅₀

Combine active influence in order further to study plasma proteins to embodiment 1 compound; A series of experiment in vitro have been carried out; Wherein this compound is derived from mouse, rat, dog, monkey or people's the serum 50% and cultivate in advance, apply it to then in A375M cell or the Mo7e cell.After cultivating a night, measure phosphoric acid-MEK and phosphoric acid-ERK level (so that mutant B-Raf is suppressed to analyze) in the A375M cell.In cultivation back 4 hours, measure the phosphoric acid-c-Kit level (so that c-Kit is suppressed to analyze) in the Mo7e cell.The result who has summarized these tests in the table 10 below.

Table 10: derive from of the active influence of the serum of various kinds to embodiment 1 compound

Can with the data in the table 10 compound of estimating embodiment 1 and the plasma proteins that derives from different plant species relative combine and with it as the basis that is used for the target plasma concns that in the mouse body, records is extrapolated to the correction factor of other kind.For example, according to these data, can estimate the intravital target plasma concns of rat doubly, and the intravital target plasma concns of people than low about 10-in the mouse body doubly than low about 5-in the mouse body.

Though the preferred embodiment of the invention is illustrated and describes, should recognize and under the situation that does not break away from purport of the present invention and scope, to carry out various variations to it.

Claims

1. the pharmaceutically useful salt of the compound of formula (I) or its tautomer or this compound or tautomer:

Wherein,

Each R ¹Be independently selected from hydroxyl, halogen, C _1-6Alkyl, C _1-6Alkoxyl group, (C _1-6Alkyl) sulfenyl, (C _1-6Alkyl) alkylsulfonyl, C _3-8Naphthenic base, Heterocyclylalkyl, phenyl and heteroaryl;

R ²Be C _1-6Alkyl or halo (C _1-6Alkyl);

Each R ⁴Be independently selected from hydroxyl, C _1-6Alkyl, C _1-6Alkoxyl group, halogen, carboxyl, (C _1-6Alkoxyl group) carbonyl, aminocarboxyl, C _1-6Alkyl amino-carbonyl, cyanic acid, C _3-8Naphthenic base, bar naphthenic base, Heterocyclylalkyl carbonyl, phenyl and heteroaryl;

A is 1,2,3,4 or 5;

B is 0,1,2 or 3; And

C is 1 or 2,

Wherein said " Heterocyclylalkyl " refers to 3 to 8 the ring carbon atom naphthenic substituents that have that in ring structure, have 1 to 2 ring hetero atom that is selected from nitrogen and oxygen; And

Described " heteroaryl " is selected from pyrazinyl, pyrazolyl, pyridyl, pyridazinyl, pyrimidyl, pyrryl and imidazolyl.

2. compound as claimed in claim 1, wherein each R ¹Be independently selected from hydroxyl, chlorine, fluorine, bromine, methyl, ethyl, propyl group, butyl, methoxyl group, oxyethyl group, propoxy-, butoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoro ethoxy, piperidyl, C _1-6Alkylpiperidine base, piperazinyl, C _1-6Alkylpiperazine base, tetrahydrofuran base, pyridyl and pyrimidyl.

3. compound as claimed in claim 1, wherein a is 1 or 2, and at least one R ¹Be halo (C _1-6Alkyl).

4. compound as claimed in claim 3, wherein at least one R ¹It is trifluoromethyl.

5. compound as claimed in claim 1, wherein R ²Be C _1-6Alkyl.

6. compound as claimed in claim 1, wherein R ²Be methyl or ethyl.

7. compound as claimed in claim 4, wherein R ²It is methyl.

8. compound as claimed in claim 1, wherein b is 0 and R ³Do not exist.

9. compound as claimed in claim 1, wherein b is 1 and R ³Be C _1-6Alkoxyl group.

10. compound as claimed in claim 9, wherein R ³It is methoxyl group.

11. compound as claimed in claim 1, wherein c is 1 or 2 and at least one R ⁴Be halo (C _1-6Alkyl).

12. compound as claimed in claim 11, wherein at least one R ⁴It is trifluoromethyl.

13. have the compound as claimed in claim 1 of formula (II) or the pharmaceutically useful salt of its tautomer or this compound or tautomer:

Wherein,

Each R ¹Be independently selected from C _1-6Alkyl, C _1-6Alkoxyl group, hydroxyl, halogen, (C _1-6Alkyl) sulfenyl, (C _1-6Alkyl) alkylsulfonyl, C _3-8Naphthenic base, Heterocyclylalkyl, phenyl and heteroaryl;

Each R ⁴Be independently selected from hydroxyl, C _1-6Alkyl, C _1-6Alkoxyl group, halogen, carboxyl, (C _1-6Alkoxyl group) carbonyl, aminocarboxyl, cyanic acid, C _3-8Naphthenic base, Heterocyclylalkyl, Heterocyclylalkyl carbonyl, phenyl and heteroaryl;

A is 1,2,3,4 or 5;

B is 0,1,2 or 3; And

C is 1 or 2,

14. have the compound as claimed in claim 1 of formula (III) or the pharmaceutically useful salt of its tautomer or this compound or tautomer:

Wherein,

A is 1,2,3,4 or 5; And

C is 1 or 2,

15. compound as claimed in claim 14, wherein each R ¹Be independently selected from hydroxyl, chlorine, fluorine, bromine, methyl, ethyl, propyl group, butyl, methoxyl group, oxyethyl group, propoxy-, butoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoro ethoxy, piperidyl, C _1-6Alkylpiperidine base, piperazinyl, C _1-6Alkylpiperazine base, tetrahydrofuran base, pyridyl and pyrimidyl.

16. compound as claimed in claim 15, wherein a is 1 or 2 and at least one R ¹Be halo (C _1-6Alkyl).

17. compound as claimed in claim 16, wherein at least one R ¹It is trifluoromethyl.

18. compound as claimed in claim 14, wherein a is 1.

19. compound as claimed in claim 18, wherein R ¹It is trifluoromethyl.

20. compound as claimed in claim 14, wherein c is 1 or 2 and at least one R ⁴Be halo (C _1-6Alkyl).

21. compound as claimed in claim 20, wherein at least one R ⁴It is trifluoromethyl.

22. compound as claimed in claim 21, wherein c is 1.

23. compound as claimed in claim 1, it is selected from following compound:

Or the pharmaceutically useful salt of its tautomer or this compound or tautomer.

24. compound as claimed in claim 1 or its pharmaceutically useful salt, wherein this compound has following formula:

25. one kind comprises like claim 1 or the pharmaceutically useful salt of 24 described compounds, tautomer, its pharmaceutically useful salt or its tautomer and the compsn of pharmaceutically acceptable carrier.

26. the application of pharmaceutically useful salt in the medicine of preparation treatment cancer like claim 1 or 24 described compounds, tautomer, its pharmaceutically useful salt or its tautomer.

27. purposes as claimed in claim 26, wherein said medicine have suppressed Raf kinases or mutant B-Raf kinases.

28. purposes as claimed in claim 26, wherein said carcinous illness is a melanoma.

29. purposes as claimed in claim 26, wherein said carcinous illness is breast cancer or prostate cancer.

30. the purposes of pharmaceutically useful salt in the medicine that preparation is used for suppressing the serine/threonine kinase of at least a MAPK signal transduction pathway in the individual's body or treating the biology situation that is mediated by the serine/threonine kinase in the MAPK signal transduction pathway in the individual's body like claim 1 or 24 described compounds, tautomer, its pharmaceutically useful salt or its tautomer.

31. purposes as claimed in claim 30, wherein said compsn suppresses the Raf kinases.

32. purposes as claimed in claim 30, wherein said compsn mutation inhibiting type B-Raf kinases.

33. purposes as claimed in claim 30, wherein said biology situation is selected from melanoma, papillary thyroid carcinoma, ovarian cancer, colorectal carcinoma, carcinoma of the pancreas, lung cancer and white blood disease.

34. purposes as claimed in claim 30, wherein said compound are selected from the pharmaceutically useful salt of { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl)-amine or its pharmaceutically useful salt, its tautomer or its tautomer.

35. be used for suppressing the individual purposes by the medicine of the biology situation of receptor tyrosine kinase mediation of individual intravital receptor tyrosine kinase or treatment like the pharmaceutically useful salt of claim 1 or 24 described compounds, tautomer, its pharmaceutically useful salt or its tautomer in preparation, wherein said receptor tyrosine kinase is selected from VEGFR-2, FGFR-3, c-Kit, PDGFR-β and CSF-1R.

36. purposes as claimed in claim 35, wherein said biology situation is selected from mast cell leukemia, erythroleukemia, germinoma, small cell lung cancer, gastrointestinal stromal tumor, acute myelogenous leukemia, neuroblastoma, melanoma, multiple myeloma, ovarian cancer and breast cancer.

37. purposes as claimed in claim 35, wherein said compound are selected from the pharmaceutically useful salt of { 1-methyl-5-[2-(5-Trifluoromethyl-1 H-imidazoles-2-yl)-pyridin-4-yl oxygen base]-1H-benzimidazolyl-2 radicals-yl }-(4-trifluoromethyl)-amine or its pharmaceutically useful salt, its tautomer or its tautomer.