CN101326186A - Pyrazolo[1,5-a]pyridine-3-carboxylic acids as EphB and VEGFR2 kinase inhibitors - Google Patents

Abstract

The invention relates to novel pyrazolo[1,5-a]pyridine-3-carboxylic acid compounds of the Formula (I) in which all of the variables are as defined in the specification, in free form or in salt form, to their preparation, to their use as medicaments and to medicaments comprising them.

Description

Pyrazolo [1,5-a] Nicotinicum Acidum compounds as EphB and VEGFR2 kinase inhibitor

The present invention relates to pyrazolo [1,5-a] the Nicotinicum Acidum compounds, it is used for the treatment of regulates the application of the disease that response is arranged or is used for the treatment of application in the pharmaceutical preparation of described disease in preparation protein kinase, the pharmaceutical preparation that comprises described compound and pharmaceutically acceptable carrier, pharmaceutical preparation in particular for described disease, be used for the treatment of animal or human's body, in particular for treating the described compound of described disease, the method of treatment animal or human body, it comprises to the animal or human uses described compound, and the method for preparing described compound, wherein in mentioning the various situations of compound, the compound of mentioning can exist with the form of this compound itself and/or salt (preferred pharmacologically acceptable salt).

Term " protein kinase " has defined the albumen that a class has enzymic activity, wherein can be divided into receptor type kinases and non-receptor type kinases and tyrosine and serine/threonine kinase.With regard to its residing position, can be divided into nuclear, tenuigenin and film associated kinase.The relevant Tyrosylprotein kinase of many films also is the acceptor of somatomedin simultaneously.

With regard to its catalytic activity, protein kinase (PK) is the enzyme of the phosphorylation of the specific Serine of catalysis intracellular protein, Threonine or tyrosine residues.The posttranslational modification of this substrate protein is carried out with the form of molecular switch usually, and it has represented a kind of step of regulating cell proliferation, activation and/or differentiation.Observed unusually in the some diseases state or excessively or more generally be inappropriate PK activity, described morbid state comprises optimum and the malignant proliferation illness.In many cases, can come to carry out in the body and external treatment by utilizing the PK inhibitor to disease.

Past has been understood Eph receptor tyrosine kinase and part thereof---the basic role of Ephrins over several years.Some different Eph acceptors have been carried out catalogue and it has been grouped into EphA or EphB subclass according to its avidity to acceptor.Identified that at least 8 kinds is the Ephrins of membranin, it is glyceryl phosphatide acyl inositol (GPI)-connecting-type (ephrinA) or strides film (ephrinB) type.As if the signal transduction between Eph acceptor and the part thereof be confined to the directly position of contact of cell-cell.The result of contact has induced some mutual two-way incidents of iuntercellular.Think that Ephrins and acceptor thereof are influential to the enterprise schemaization (patterning) and the group structure of the very limited cell position in space in the expression at some position.The concrete influence of wherein some comprises that changing cell migration, adhesion and body segment forms.

EphB4 (being also referred to as HTK) and part ephrinB2 (HTKL) thereof play an important role in the foundation of blood vessel network with in determining.EphB4 is specific expressed on the vein epithelium, and early stage in vascularization, and ephrinB2 is specificity and expression on the contrary on arterial endothelial cell.The gene of dysfunction has caused the lethality of mice embryonic, and in ephrinB2 or the defective situation of EphB4, the embryo is showing identical defective aspect the connection of formation capillary vessel.Between the emergence period, the two is all expressed on first position of hemopoietic and blood vessel foundation the embryo.Determined that it plays an important role for suitable hematopoiesis, endothelium, angioblast and primitive mesoblast growth.EphB4 lacks makes embryonic stem cell mesoderm differentiated result be changed.The ectopic expression of EphB4 in breast tissue makes that udder texture system confusion, function of organization are unusual and the malignization tendency arranged (referring to people such as for example N.Munarini, J.Cell.Sci. 115, 25-37 (2002)).By these data and other data, infer that may relate to inappropriate EphB4 in malignant tumour forms expresses, therefore, it is to anti-malignant tumor a kind of means of cancer etc. for example that expection suppresses EphB4.

The viral form c-Src of the constitutive expression of the Tyrosylprotein kinase c-Src that finds in cell (deriving from Rous Sarcoma virus, a kind of retrovirus) is how the incorrect expression of Src protein tyrosine kinase causes an example based on the malignant tumour of cell transformed.Suppress the Src protein tyrosine kinase and make the growth out of control of the tumour cell that can suppress the conversion in the connective tissue tumor for example.Therefore, expection inhibition c-Src or its modification or mutant form have useful influence to the treatment of proliferative disease.

Known VEGFR (vascular endothelial growth factor receptor) participated in vasculogenesis from the beginning of control.Because solid tumor especially depends on good blood supply, therefore, in such tumor treatment, also suppressing vasculogenesis thus carries out clinical study to suppressing VEGFR, and has shown some results likely.VEGF also is main participant in leukemia and lymphoma, and it is expressed at many malignant solid tumor camber, gets in touch closely with the process of malignant disease.The example of the tumour of VEGF expression R-2 (KDR) has lung cancer, breast cancer, non_hodgkin lymphoma, ovarian cancer, carcinoma of the pancreas, malignant pleural mesothelioma and melanoma.Except that its angiogenic activity, the part of VEGFR---VEGF also can promote tumor growth by directly short survival (pro-survival) effect to tumour cell.Also have numerous disease and associated angiogenesis out of control, for example following disease.

In the patient who suffers from chronic granulocytic leukemia (CML), observe the abl proto-oncogene and change into oncogene.Chromosome translocation is connected to the bcr gene on the chromosome 22 on the abl gene of karyomit(e) 9, thereby has produced Philadelphia chromosome.The fusion rotein that is produced has the proteic N-terminal of Bcr that links to each other with the C-terminal of Abl tyrosine protein kinase.As a result, the Abl kinase domain has been driven hematopoietic cell clone's hyper-proliferative in the marrow by unsuitable activation.Shown and used activeconstituents Gleevec ^TMOr Glivec (trade mark of Novartis, the inhibitor of this fusion rotein) suppresses this Tyrosylprotein kinase is highly effective CML therapy.Therefore, the insufficient expression that can verify the Abl Tyrosylprotein kinase can be treated malignant tumour, especially leukemic general concept.

But, up to now, as the chemical compound lot of kinases inhibitor lack specificity, have undesirable side effect (this side effect is because the rejection characteristic of disadvantageous protein kinase to more than one types causes), lack effectiveness owing to specificity is too high, only effective to some disease, during administration generation tolerance and/or similar inadvisable character.

Therefore, problem to be solved by this invention is: owing to a large amount of kinases inhibitors and multiple proliferative disease and other disease relevant with protein kinase are arranged and because some therapy is formed tolerance, therefore need to provide to can be used as kinases inhibitor and so can be used for treating these and the new compound type of protein tyrosine kinase such as serine/threonine and/or the disease that preferably PTK (protein tyrosine kinase) is relevant always.Required is pharmaceutically favourable arrestin kinases, especially the new compound of PTK, its especially have favorable properties as to limited group or even single protein kinase has high affinity and/or selectivity, in to the situation of dissimilar compounds tolerances effectively, the kinases of particular group is had useful affinity.That is, need to address the above problem or the new kinases inhibitor of other problem.

Described Hydrazonopyrazole miazines compound that some 4-replaces and can be used as the GSK3 kinase inhibitor that is used for the treatment of diabetes and the disease relevant, seen WO04/009602, WO 04/009596 or WO 04/009597 with the TIE-2 kinases.On the other hand, some acyl groups as the p38-inhibitor-or the arylamino-Pyrazolopyrimidines of acyl amino-replacement has been described, referring to WO 03/099280.

General description of the present invention

Now find surprisingly, pyrazolo [1 of the present invention, 5-a] the Nicotinicum Acidum compound can be used for suppressing many and may participate in by the signal transmission of nutritional factor mediation and relate to the protein kinases of the disease performance (for example proliferative (for example tumour) growth) of this protein kinase activity, especially the representative instance of protein tyrosine kinase, especially be selected from following kinases family: the src kinases, especially c-src kinases, VEGF-receptor kinase (for example KDR and Flt-1), RET-receptor kinase and/or Ephrin receptor kinase, EphB2 kinases for example, EphB4 kinases or associated kinase, in addition, also has the abl kinases, especially v-abl or c-abl kinases, b-raf (V599E), other kinases of EGF receptor kinase or EGF family, for example HER-1 or c-erbB2 kinases (HER-2), Flt-3, Ick, fyn, the c-erbB3 kinases, the c-erbB4 kinases; The member of PDGF-receptor tyrosine protein kinase family, PDGF-receptor kinase for example, the CSF-1 receptor kinase, Kit-receptor kinase (c-Kit), the FGF-receptor kinase, FGF-R1 for example, FGF-R2, FGF-R3, FGF-R4, c-Raf, casein kinase (CK-1, CK-2, G-CK), Pak, ALK, ZAP70, Jak1, Jak2, Axl, Cdk1, cdk4, cdk5, Met, FAK, Pyk2, Syk, Tie-2, insulin receptor kinase (Ins-R), the receptor kinase of rhIGF-1 (IGF-1 kinases) and/or other serine/threonine kinases, protein kinase C (PK-C) for example, PK-B, EK-B or cdc kinases are as CDK1, and any or multiple sudden change in these kinases (for example forming activation) form (Bcr-Abl for example, RET/MEN2A, RET/MEN2B, RET/PTC1-9 or b-raf (V599E)).All these with other protein kinase all in the growth regulating of the mammalian cell that comprises people's cell with play a part certain in transforming.Especially find that pair cell Eph4B kinases is very effective.

Because these activity, compound of the present invention can be used for treating regulates the disease that response is arranged to protein kinase, for example especially with such kinase whose activity unusual (for example nonadjustable, out of control or composition activity) or active excessively diseases associated, especially those diseases of being mentioned, and preferred those diseases of especially being mentioned.

Detailed description of the present invention

In first embodiment, the present invention relates to the compound of the formula (I) of free or salt form

Wherein

R1 is hydrogen, halogen, sulfamyl, N-C _1-4Alkylsulfamoyl group, N, N-two-C _1-4Alkylsulfamoyl group, be not substituted or substituted alkyl, be not substituted or substituted aryl, be not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical;

R2 and R3 are hydrogen, halogen, C independently of one another _1-4-alkyl, trifluoromethyl, C _1-4-alkoxyl group or cyano group,

X ₁, X ₂, X ₃And X ₄Be that CH or wherein maximum two can be N;

D is N (R6) (preferably), O or S,

Wherein R6 is hydrogen, acyl group or is not substituted or substituted alkyl,

Y ₁Be O, S, NH, CH ₂, N=CH, CH=N or CH=CH;

Y ₂Be C or, if Ra does not exist, then its (also) can be N;

Ra is

If-Y ₂Be that N and R1 are unsubstituted alkyl, substituted alkyl with 5 or more carbon atoms, are not substituted or substituted aryl, are not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical that it does not exist, perhaps,

If-Y ₂Being that C and R1 are unsubstituted alkyl, substituted alkyl with 5 or more carbon atoms, are not substituted or substituted aryl, are not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical, is hydrogen,

If Y ₂Be that C and R1 are hydrogen, halogen, C _1-4-alkyl, sulfamyl, N-C _1-4Alkylsulfamoyl group, N, N-two-C _1-4Alkylsulfamoyl group or unsubstituted aryl are the parts of formula IA

Wherein dotted line is represented and the rest part of formula I molecule (with shown in the wavy line) bonded key,

A be C (=O)-N (R4) or N (R4)-C (=O),

Wherein R4 is hydrogen or is not substituted or substituted alkyl,

Z ₁Be O, S, NH, CH ₂, CH=N, N=CH or CH=CH,

Z ₂Be nitrogen or CH,

Z ₃Be CH or N,

Each R that occurs ₅Be a kind of substituting group independently of one another, and

N is 0,1 or 2.

In another embodiment, the present invention relates to the formula I compound of following free form or salt form, wherein

R1 is hydrogen, halogen, sulfamyl, N-C _1-4Alkylsulfamoyl group, N, N-two-C _1-4Alkylsulfamoyl group, be not substituted or substituted alkyl, be not substituted or substituted aryl or be not substituted or substituted heterocyclic radical;

R2 and R3 are hydrogen or C independently of one another _1-4-alkyl,

X ₁, X ₂, X ₃And X ₄Be CH;

D is N (R6) (preferably), O or S,

Wherein R6 is hydrogen, acyl group or is not substituted or substituted alkyl,

Y ₁Be O, S, NH, CH ₂, N=CH, CH=N or CH=CH;

Y ₂Be C;

Ra is

If-R1 is unsubstituted alkyl, substituted alkyl with 5 or more carbon atoms, be not substituted or substituted aryl, be not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical, be hydrogen,

-or, if R1 is hydrogen, halogen, C _1-4-alkyl, sulfamyl, N-C _1-4Alkylsulfamoyl group, N, N-two-C _1-4Alkylsulfamoyl group, being substituted or unsubstituted aryl, is the part of formula IA,

A be C (=O)-N (R4) or N (R4)-C (=O),

Wherein R4 is hydrogen or is not substituted or substituted alkyl,

Z ₁Be O, S, NH, CH ₂, CH=N, N=CH or CH=CH,

Z ₂Be nitrogen or CH,

Z ₃Be CH or N,

Each R that occurs ₅Be a kind of substituting group independently of one another, and

N is 0,1 or 2.

In another embodiment, the present invention relates to the formula I compound of following free form or salt form, wherein

R1 is hydrogen, N, N-two-C _1-4Alkylsulfamoyl group, be not substituted or substituted aryl;

R2 and R3 are hydrogen or C independently of one another _1-4-alkyl,

X ₁, X ₂, X ₃And X ₄Be that CH or wherein maximum two can be N;

D is N (R6) (preferably), O or S,

Wherein R6 is hydrogen, acyl group or is not substituted or substituted alkyl,

Y ₁Be O, S, NH, CH ₂, N=CH, CH=N or CH=CH;

Y ₂Be C;

Ra is

If-R1 is unsubstituted alkyl, substituted alkyl with 5 or more carbon atoms, be not substituted or substituted aryl, be not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical, be hydrogen,

-or, if R1 is hydrogen, N, N-two-C _1-4Alkylsulfamoyl group or be not substituted or substituted aryl is the part of formula IA,

A be C (=O)-N (R4) or N (R4)-C (=O),

Wherein R4 is hydrogen or is not substituted or substituted alkyl,

Z ₁Be O, S, NH, CH ₂, CH=N, N=CH or CH=CH,

Z ₂Be nitrogen or CH,

Z ₃Be CH or N,

Each R that occurs ₅Be a kind of substituting group independently of one another, and

N is 0,1 or 2.

In another embodiment, the present invention relates to the formula I compound of following free form or salt form, wherein

R1 is hydrogen, N, N-two-C _1-4Alkylsulfamoyl group, be not substituted or substituted phenyl;

R2 and R3 are hydrogen or C independently of one another _1-4-alkyl,

X ₁, X ₂, X ₃And X ₄Be that CH or wherein maximum two can be N;

D is N (R6) (preferably), O or S,

Wherein R6 is hydrogen, acyl group or is not substituted or substituted alkyl,

Y ₁Be O, S, NH, CH ₂, N=CH, CH=N or CH=CH;

Y ₂Be C;

Ra is

If-R1 is not substituted or substituted phenyl, is hydrogen,

-or, if R1 is hydrogen, N, N-two-C _1-4Alkylsulfamoyl group or unsubstituted phenyl are the parts of formula IA,

A be C (=O)-N (R4) or N (R4)-C (=O),

Wherein R4 is hydrogen or is not substituted or substituted alkyl,

Z ₁Be O, S, NH, CH ₂, CH=N, N=CH or CH=CH,

Z ₂Be nitrogen or CH,

Z ₃Be CH or N,

Each R that occurs ₅Be a kind of substituting group independently of one another, and

N is 0,1 or 2.

In another embodiment, the present invention relates to the formula I compound of following free form or salt form, wherein

R1 is hydrogen, N, N-two-C _1-4Alkylsulfamoyl group, be not substituted or substituted phenyl;

R2 and R3 are hydrogen or C independently of one another _1-4-alkyl,

X ₁, X ₂, X ₃And X ₄Be CH;

D is NH,

Y ₁Be CH=CH;

Y ₂Be C;

Ra is

If-R1 is substituted phenyl, be hydrogen,

-or, if R1 is hydrogen, N, N-two-C _1-4Alkylsulfamoyl group or unsubstituted phenyl are the parts of formula IA,

A be C (=O)-N (R4) or N (R4)-C (=O),

Wherein R4 is hydrogen or is not substituted or substituted alkyl,

Z ₁Be O, S, NH, CH ₂, CH=N, N=CH or CH=CH,

Z ₂Be nitrogen or CH,

Z ₃Be CH or N,

Each R that occurs ₅Be a kind of substituting group independently of one another, and

N is 0,1 or 2.

In another embodiment, the present invention relates to the formula I compound of following free form or salt form, wherein

R1 is hydrogen, N, N-two-C _1-4Alkylsulfamoyl group, be not substituted or substituted phenyl,

R2 and R3 are hydrogen or C independently of one another _1-4-alkyl,

X ₁, X ₂, X ₃And X ₄Be CH;

D is NH,

Y ₁Be CH=CH;

Y ₂Be C;

Ra is

If-R1 is substituted phenyl, be hydrogen,

-or, if R1 is hydrogen, N, N-two-C _1-4Alkylsulfamoyl group or unsubstituted phenyl are the parts of formula IA,

A be C (=O)-NH or NH-C (=O),

Z ₁Be CH=CH,

Z ₂Be CH,

Z ₃Be CH,

Each R that occurs ₅Be a kind of substituting group independently of one another, and

N is 0,1 or 2.

In another embodiment, the present invention relates to the formula I compound of following free form or salt form, wherein

R1 is hydrogen or substituted phenyl,

R2 and R3 are hydrogen or C independently of one another _1-4-alkyl,

X ₁, X ₂, X ₃And X ₄Be CH;

D is NH,

Y ₁Be CH=CH;

Y ₂Be C;

Ra is

If-R1 is substituted phenyl, be hydrogen,

If R1 is hydrogen or unsubstituted phenyl, be the part of formula IA,

A be C (=O)-NH or NH-C (=O),

Z ₁Be CH=CH,

Z ₂Be CH,

Z ₃Be CH,

Each R that occurs ₅Be a kind of substituting group independently of one another, and

N is 0,1 or 2.

The compound or its pharmaceutically useful salt that the present invention relates to formula I are used for the treatment of the application of protein kinase being regulated the disease that response is arranged, described disease is animal or preferred people's disease especially, especially the disease that the inhibition of described protein tyrosine kinase (PTK) under one or more " general descriptions of the present invention " is had response, described kinases more particularly is selected from one or more PTK:src kinases of following kinases family, especially c-src kinases, VEGF-receptor kinase (for example KDR and Flt-1), RET-receptor kinase or Ephrin receptor kinase, for example EphB2 kinases, EphB4 kinases or associated kinase, or its sudden change (for example forming activation or partially or completely out of control) form.

The invention still further relates to the compound of formula I or its (preferably pharmaceutically useful) salt and be used for the treatment of application in the pharmaceutical preparation of described disease in preparation; Comprise the compound of formula I or the pharmaceutical preparation of its pharmaceutically useful salt and at least a pharmaceutically acceptable carrier, in particular for treating the pharmaceutical preparation of described disease; Be used for the treatment of animal or human's body, in particular for compound or its pharmaceutically useful salt of the formula I of treatment earlier paragraphs described disease; The method of treatment animal or human body, it comprises to the animal or human, especially needs compound or its pharmaceutically useful salt of formula I of the described disease significant quantity of animal or human's administering therapeutic of such treatment; And the method for the compound of preparation formula I or its (preferably pharmaceutically useful) salt.

In formula I, independent, set or close the implication of form below preferred with its any combination or subgroup:

In this article, unless stated otherwise, otherwise used general terms or symbol has following implication in the context:

Term " rudimentary " or " C ₁-C ₇-" defined a kind of have high to and comprise 7 at most, especially high to and comprise the part of 4 carbon atoms at most, described part is side chain (branch's one or many) or straight chain.Rudimentary or C ₁-C ₇-alkyl for example be just-amyl group, just-hexyl or just-heptyl or preferred C ₁-C ₄-alkyl, especially methyl, ethyl, just-propyl group, the second month in a season-propyl group, just-butyl, isobutyl-, the second month in a season-butyl, tert-butyl.In the situation of low-grade alkenyl or low-grade alkynyl, the rudimentary " C that preferably refers to ₂-C ₇"-, perhaps more preferably refers to " C ₂-C ₄-".

Halo or halogen be fluorine, chlorine, bromine or iodine preferably, most preferably is fluorine, chlorine or bromine.

Be not substituted or substituted alkyl C preferably ₁-C ₂₀-alkyl, it more preferably is low alkyl group, methyl for example, ethyl or propyl group, its can be straight chain or can branch's one or many (prerequisite is that carbonatoms allows) and be not substituted or by one or more, preferred maximum three are independently selected from following substituting group and replace: be not substituted or substituted as described as follows heterocyclic radical, especially tetramethyleneimine-1-base, piperidines-1-base, by amino or N-single-or N, N-two-[low alkyl group, phenyl and/or phenyl-low alkyl group]-the amino piperidines-1-base that replaces, by the piperidyl that the ring carbon atom bonded is not substituted or the N-low alkyl group replaces, as 1-sec.-propyl-piperidin-4-yl, piperazine-1-base, low alkyl group piperazine-1-base is as 4-(methyl, ethyl or sec.-propyl)-piperazine-1-base, morpholine-4-base or thiomorpholine-4-base; Be not substituted or as following defined substituted cycloalkyl, be not substituted or as following defined substituted aryl, especially phenyl or naphthyl; Or low-grade alkenyl, low-grade alkynyl, halogen, hydroxyl, lower alkoxy, rudimentary-alkoxyl group-lower alkoxy, (rudimentary-alkoxyl group)-lower alkoxy-lower alkoxy, phenoxy group, naphthyloxy, phenyl-or naphthyl-lower alkoxy, as benzyloxy; Amino-lower alkoxy, rudimentary-alkanoyloxy, benzoyloxy, naphthoyl oxygen base, nitro, cyano group, carboxyl, elementary alkoxy carbonyl, for example methoxycarbonyl, just-propoxycarbonyl, different-propoxycarbonyl or uncle-butoxy carbonyl; Phenyl-or naphthyl-elementary alkoxy carbonyl, as benzyloxycarbonyl; Low-grade alkane acidyl, as ethanoyl, benzoyl, naphthoyl, formamyl, N-single-or N, the dibasic formamyl of N-, as N-single-or N, the dibasic formamyl of N-, wherein said substituting group is selected from low alkyl group and hydroxy lower alkyl; Amidino groups, guanidine radicals, urea groups, sulfydryl, lower alkylthio, thiophenyl or naphthalene sulfenyl, phenyl-or naphthyl-lower alkylthio, low alkyl group-thiophenyl, low alkyl group-naphthalene sulfenyl, halo-low alkyl group sulfydryl, the low alkyl group sulfinyl, phenyl-or naphthyl-sulfinyl, phenyl-or naphthyl-low alkyl group sulfinyl, low alkyl group-phenyl sulfinyl, low alkyl group-naphthyl sulfinyl, sulfo group, the lower alkane alkylsulfonyl, phenyl-or naphthyl-alkylsulfonyl, phenyl-or naphthyl-low alkyl group alkylsulfonyl, the low alkyl group phenyl sulfonyl, halo-low alkyl group alkylsulfonyl is as trifyl; Sulfonamido, phenylsulfonamido, amino, N-be single-or N, N-two-[low alkyl group, phenyl and/or phenyl-low alkyl group]-amino, as N, N-dimethylamino, N, N-diethyl-amino, 3-[N-(N, the N-dimethylamino)-propyl group amino, 2-[N-(N, N-dimethylamino)-ethylamino or N-(N, N-dimethylamino)-methylamino; Wherein as substituting group or above-mentioned each phenyl or naphthyl (yet comprising the phenyl or naphthyl that is arranged in phenoxy group or naphthyloxy) of being substituted the substituent part of alkyl is not substituted itself or by one or more; for example maximum 3; preferred 1 or 2 is independently selected from following substituting group and replaces: halogen; especially fluorine; chlorine; bromine or iodine; halo-low alkyl group; as trifluoromethyl; hydroxyl; lower alkoxy; amino; the N-list-or N, N-two-(low alkyl group; phenyl; naphthyl; phenyl-low alkyl group and/or naphthyl-low alkyl group)-amino; nitro; carboxyl; rudimentary-alkoxy carbonyl; formamyl; cyano group and/or sulfamyl.R1 among the formula I is low alkyl group especially preferably, amino-low alkyl group, as the 3-aminopropyl, 2-amino-ethyl or 2-amino methyl, the N-list-or N, N-two-(low alkyl group, phenyl and/or phenyl-low alkyl group)-amino-low alkyl group, as 3-(N, the N-dimethylamino)-propyl group, 3-(N, the N-diethylamino)-propyl group, 2-(N, the N-dimethylamino)-ethyl, 2-(N, the N-diethylamino)-ethyl, N, N-dimethylaminomethyl or N, N-diethylamino methyl, tetramethyleneimine-1-base-low alkyl group, piperidines-1-base-low alkyl group, 1-low alkyl group piperidin-4-yl-low alkyl group, the 4-[N-list-or N, N-two-(low alkyl group, phenyl and/or phenyl-low alkyl group)-amino]-piperidines-1-base, piperazine-1-base-low alkyl group, as piperazine-1-base-methyl, 4-low alkyl group piperazine-1-base-low alkyl group is as 4-(methyl, ethyl or sec.-propyl)-piperazine-1-base-methyl or (morpholine-4-base or thiomorpholine-4-yl)-low alkyl group.Have 5 or the alkyl of more a plurality of carbon atoms C especially ₅-C ₂₀-alkyl.Find that the Ra in formula I does not exist or (if Y especially with regard to regard to the organized enzyme conformation combines ₂Be C) R during hydrogen ₁C preferably ₅-C ₂₀-alkyl or substituted C ₁-C ₂₀-alkyl.Perhaps, find especially with regard to regard to the enzyme conformation of non-activity combines the Y in formula I ₂Be C and Ra when being formula IA part shown in top, R ₁C preferably ₁-C ₄-alkyl or especially hydrogen.

Be not substituted or substituted aryl preferably has and is no more than 20 carbon atoms, especially be no more than the unsaturated carbon loop systems of 16 carbon atoms, it is preferably single-, two-or three rings, it is not substituted, perhaps, and in the situation of substituted aryl, it is preferably by one or more, preferred maximum three, for example one or two is independently selected from following substituting group and replaces: phenyl, naphthyl, phenyl-or naphthyl-low alkyl group, as benzyl; Hydroxy lower alkyl, as hydroxymethyl; Rudimentary-alkoxyl group-low alkyl group, (rudimentary-alkoxyl group)-lower alkoxy-low alkyl group, low-grade alkane acidyl-low alkyl group, halo-low alkyl group are as trifluoromethyl; Phenoxy group-or naphthyloxy-low alkyl group, phenyl-or naphthyl-lower alkoxy-low alkyl group, as benzyloxy-low alkyl group; Lower alkoxy-carbonyl oxygen base-low alkyl group is as uncle-butoxy carbonyl oxy-low alkyl group; Phenyl-or naphthyl-lower alkoxy carbonyl oxygen base-low alkyl group, as benzyloxy carbonyl oxygen base-low alkyl group; Cyano group-low alkyl group, low-grade alkenyl, low-grade alkynyl, low-grade alkane acidyl are as ethanoyl; Halogen, hydroxyl, lower alkoxy are as methoxyl group, rudimentary-alkoxyl group-lower alkoxy, (rudimentary-alkoxyl group)-lower alkoxy-lower alkoxy, phenoxy group, naphthyloxy, phenyl-or naphthyl-lower alkoxy, as benzyloxy; Amino-lower alkoxy, rudimentary-alkanoyloxy, benzoyloxy, naphthoyl oxygen base, nitro, amino, list-, two-or three-(in the later case by quaternized and lotus positive electricity) amino of replacing, wherein said amino substituting group is independently selected from low alkyl group, low-grade alkane acidyl, lower alkane alkylsulfonyl, as methylsulfonyl, phenyl, naphthyl, phenyl-low alkyl group and naphthyl-low alkyl group; Cyano group, carboxyl, elementary alkoxy carbonyl, for example methoxycarbonyl, just-propoxycarbonyl, different-propoxycarbonyl or uncle-butoxy carbonyl; Phenyl-or naphthyl-elementary alkoxy carbonyl, as benzyloxycarbonyl; Benzoyl, naphthoyl, formamyl, N-be single-or N, the dibasic formamyl of N-, as N-single-or N, the dibasic formamyl of N-, wherein said substituting group is selected from low alkyl group and hydroxy lower alkyl; Amidino groups, guanidine radicals, urea groups, sulfydryl, lower alkylthio, thiophenyl or naphthalene sulfenyl, phenyl-or naphthyl-lower alkylthio, low alkyl group-thiophenyl, low alkyl group-naphthalene sulfenyl, halo-low alkyl group sulfydryl, the low alkyl group sulfinyl, phenyl-or naphthyl-sulfinyl, phenyl-or naphthyl-low alkyl group sulfinyl, low alkyl group-phenyl sulfinyl, low alkyl group-naphthyl sulfinyl, sulfo group, the lower alkane alkylsulfonyl, phenyl-or naphthyl-alkylsulfonyl, phenyl-or naphthyl-low alkyl group alkylsulfonyl, the low alkyl group phenyl sulfonyl, halo-low alkyl group alkylsulfonyl is as trifyl; Sulfonamido, phenylsulfonamido, tetramethyleneimine-1-base, piperidines-1-base, by amino or N-single-or N, N-two-[low alkyl group, phenyl and/or phenyl-low alkyl group)-the amino piperidines-1-base that replaces, by the ring carbon atom bonded is not substituted or the N-low alkyl group replaces piperidyl, as 1-sec.-propyl-piperidin-4-yl, piperazine-1-base, low alkyl group piperazine-1-base, as 4-(methyl, ethyl or sec.-propyl)-piperazine-1-base, morpholine-4-base or thiomorpholine-4-base; Wherein as substituting group or above-mentioned each phenyl or naphthyl (yet comprising the phenyl or naphthyl that is arranged in phenoxy group or naphthyloxy) of being substituted the part of aryl substituent is not substituted itself or by one or more; for example maximum 3; preferred 1 or 2 is selected from following substituting group and replaces: halogen; especially fluorine; chlorine; bromine or iodine; halo-low alkyl group; as trifluoromethyl; hydroxyl; lower alkoxy; amino; the N-list-or N, N-two-(low alkyl group; phenyl; naphthyl; phenyl-low alkyl group and/or naphthyl-low alkyl group) amino; nitro; carboxyl; rudimentary-alkoxy carbonyl; formamyl; cyano group and/or sulfamyl.Be not substituted or substituted aryl (especially with regard to the R1 among the formula I) preferably is not substituted or the phenyl that replaced by following substituting group: halogen; more preferably lower alkoxy; nitro; amino; low-grade alkane acidyl amino; N-lower alkane sulfuryl amino; as methylsulfonyl amino; the N-list-; N; N-two-or N; N; N-three-(low alkyl group; phenyl and/or phenyl-low alkyl group)-amino (latter is equivalent to season amino=quaternary ammonium); tetramethyleneimine-1-base; piperidines-1-base; by amino or N-single-or N; N-two-[low alkyl group; phenyl and/or phenyl-low alkyl group)-the amino piperidines-1-base that replaces; by the piperidyl that the ring carbon atom bonded is not substituted or the N-low alkyl group replaces; as 1-sec.-propyl-piperidin-4-yl; piperazine-1-base; low alkyl group piperazine-1-base is as 4-(methyl; ethyl or sec.-propyl)-piperazine-1-base; C _1-7Alkoxyl group-carbonyl-piperazine-1-base, for example uncle-butoxy carbonyl-piperazine-1-base, cycloalkyloxy-carbonyl-piperazine-1-base, aryloxycarbonyl-piperazine-1-base, morpholine-4-base or thiomorpholine-4-base.Aryl is phenyl preferably.

Substituted aryl, for example substituted phenyl are preferably replaced by the substituting group below 1 or 2: C _1-7-alkoxyl group, for example C _1-4-alkoxyl group, morpholine-4-base, N, N-two-C _1-7-alkylamino, N for example, N-two-C _1-4-alkylamino, C _1-7Alkoxyl group-carbonyl-piperazine-1-base, for example uncle-butoxy carbonyl-piperazine-1-base, cycloalkyloxy-carbonyl-piperazine-1-base, aryloxycarbonyl-piperazine-1-base.

Be not substituted or substituted cycloalkyl in, cycloalkyl preferably has 3 to 16, the saturated mono of preferred 3 to 9 ring carbon atoms-or the bicyclic hydrocarbon base, for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl or ring octyl group, and by one or more, preferred one to three is independently selected from about the described substituting group of substituted aryl and replaces or be not substituted (being preferred).

Be not substituted or substituted heterocyclic radical is preferably unsaturated, saturated or the heterocyclic group of fractional saturation, and it being monocycle or broadly of the present invention, is two ring or trinucleated; Have 3 to 24 annular atomses, more preferably have 4 to 16 annular atomses, most preferably have 4 to 10 annular atomses; Wherein one or more, preferred one to four, especially one or two carboatomic ring atom heteroatoms of being selected from nitrogen, oxygen and sulphur replaces, and coupling collar preferably has 4 to 12, especially 5 to 7 annular atomses; This heterocyclic group (heterocyclic radical) is not substituted or by one or more, especially 1 to 3 be independently selected from above " substituted aryl " down defined substituting group replace; And this heterocyclic radical especially is selected from Oxyranyle, aziridinyl, the ethylenimine base, 1,2-oxygen thia cyclopentyl, thienyl, furyl, tetrahydrofuran base, pyranyl, the thiapyran base, thianthrenyl, isobenzofuran-base, benzofuryl, chromenyl, the 2H-pyrryl, pyrryl, pyrrolinyl, pyrrolidyl, imidazolyl, imidazolidyl, benzimidazolyl-, pyrazolyl, pyrazinyl, pyrazolidyl, thiazolyl, isothiazolyl, dithiazole base oxazolyl isoxazolyl, pyridyl, pyrazinyl, pyrimidyl, piperidyl, piperazinyl, pyridazinyl, morpholinyl, thio-morpholinyl, (S-oxo or S, the S-dioxo)-thio-morpholinyl, the indolizine base, pseudoindoyl, the 3H-indyl, indyl, benzimidazolyl-, cumaryl, indazolyl, triazolyl, tetrazyl, purine radicals, the 4H-quinolizinyl, isoquinolyl, quinolyl, tetrahydric quinoline group, tetrahydro isoquinolyl, decahydroquinolyl, the octahydro isoquinolyl, benzofuryl, dibenzofuran group, benzothienyl, the dibenzothiophene base, phthalazinyl, naphthyridinyl, quinoxalinyl (quinoxalyl), quinazolyl, quinazolyl, the cinnolines base, pteridyl, carbazyl, the β-Ka Lin base, phenanthridinyl, acridyl, perimidinyl, the phenanthroline base, the furazan base, phenazinyl, phenothiazinyl phenoxazinyl, chromenyl, different chromanyl and chromanyl, these groups are not substituted separately or are selected from low alkyl group by one or two, especially methyl or tert-butyl, lower alkoxy, especially methoxyl group, and the group of halogen, especially bromine or chlorine replaces.

X ₁, X ₂, X ₃And X ₄Preferably all be CH.

Acyl group preferably be selected from by carbonyl (C (=O)-) or alkylsulfonyl (S (=O) ₂-) group is connected to not being substituted or substituted alkyl, not being substituted or substituted aryl, not being substituted or substituted heterocyclic radical or be not substituted or the organic moiety of substituted cycloalkyl on the rest part of formula I molecule, it separately preferably as mentioned above, that is, it is a kind of part that derives from organic carboxyl acid or sulfonic acid.Preferably alkyloyl, especially low-grade alkane acidyl, for example ethanoyl, propionyl or butyryl radicals, benzoyl (=phenylcarbonyl group), naphthoyl (=naphthyl carbonyl), phenyl-C ₁-C ₇-alkyl-carbonyl, naphthyl-C ₁-C ₇-alkyl-carbonyl; phenyl sulfonyl or lower alkane alkylsulfonyl; wherein be not substituted or by one or more as each low-grade alkane acidyl of acyl group or as each phenyl or naphthyl that the part of acyl group is mentioned; for example maximum 3; preferred 1 or 2 is independently selected from following substituting group and replaces: halogen; especially fluorine; chlorine; bromine or iodine; halo-low alkyl group; as trifluoromethyl; hydroxyl; lower alkoxy; amino; the N-list-or N, N-two-(low alkyl group; phenyl; naphthyl; phenyl-low alkyl group or naphthyl-low alkyl group) amino; nitro; carboxyl; rudimentary-alkoxy carbonyl; formamyl; cyano group and/or sulfamyl.Preferred low-grade alkane acidyl, benzoyl, phenyl sulfonyl or tosyl group.

Y ₁Preferably N=CH, CH=N or most preferably be CH=CH.

Y ₂Be that C (carbon) or (if Ra does not exist) can be N, it is meant Y ₂-Ra can be C (Ra) or N, Y ₂C preferably.

In the formula IA part in the above, " dotted line is represented rest part (shown in wavy line) the bonded key with formula I molecule " is meant that dotted line is equivalent to the key of the Ra of combined type I.

Be not substituted or substituted alkyl R4 can be as defined above not being substituted or substituted alkyl; Preferably low alkyl group or phenyl-low alkyl group.

Z ₁Preferably CH=N or N=CH or more preferably be CH=CH (forming a kind of six-ring thus).

Z ₂And Z ₃CH preferably separately.

Above substituent R 5 (if present, that is, if n is 1 or 2) preferably is independently selected from about the described substituting group of substituted aryl; Especially be selected from low alkyl group, halogen, halo-low alkyl group, as the trifluoromethyl low-grade alkane acidyl, as ethanoyl; Hydroxyl, lower alkoxy, as the amino of methoxyl group, nitro, amino, list-or two-replace, wherein amino substituting group is independently selected from low alkyl group and low-grade alkane acidyl, cyano group, carboxyl, elementary alkoxy carbonyl.For example methoxycarbonyl, formamyl, N-single-or N, formamyl, amidino groups, guanidine radicals, urea groups, lower alkylthio, sulfo group, lower alkane alkylsulfonyl and the sulfonamido of N-two-(low alkyl group)-replacement.R5 is C preferably _1-7-alkoxyl group more preferably is C _1-4-alkoxyl group.

Symbol n preferably represents 0 or 1.

Salt is the pharmacologically acceptable salt of formula I compound especially.Have salt forming group, in the situation as alkalescence or acidic-group, can form these salt, can there be (for example in pH is 4 to 10 aquosity environment) down to the dissociated form of small part in it, perhaps especially can be separated with solid form; Perhaps, in the situation that has charged group (for example quaternary ammonium), also can form these salt-in the later case, with organic acid or mineral acid (such as in the hypomere definition) anion forming acids salt.

Such salt for example can be formed the acid salt form, the preferred salt that is formed by the formula I compound with basic nitrogen atom and organic acid or mineral acid, especially pharmaceutically useful salt.Suitable mineral acid has for example haloid acid, example hydrochloric acid, sulfuric acid or phosphoric acid.Suitable organic acid has for example carboxylic acid, phosphonic acids, sulfonic acid or thionamic acid, for example acetate, propionic acid, lactic acid, fumaric acid, succsinic acid, citric acid, amino acid, as L-glutamic acid or aspartic acid, toxilic acid, hydroxymaleic acid, methyl-maleic acid, phenylformic acid, methylsulfonic acid or ethyl sulfonic acid, ethane-1,2-disulfonic acid, Phenylsulfonic acid, 2-naphthene sulfonic acid, 1,5-naphthalene-disulfonic acid, N-cyclohexyl thionamic acid, N-methyl-, N-ethyl-or N-propyl group-thionamic acid, or other organic protonic acid, as xitix.

In the situation that has bear electric group such as carboxyl or sulfo group, can also be the salt that forms with alkali, as metal or ammonium salt, as basic metal or alkaline earth salt, for example sodium, potassium, magnesium or calcium salt, or with ammonia or suitable organic amine such as monobasic tertiary amine, triethylamine or three (2-hydroxyethyl) amine for example, or heterocyclic bases, for example N-ethyl-piperidines or N, the ammonium salt that N '-lupetazin forms.

When having basic group and acidic-group simultaneously in a part, the compound of formula I can also form inner salt.

For isolated or purified, can also use pharmaceutically unacceptable salt, for example picrate or perchlorate.For treatment is used, only use pharmaceutically useful salt or free cpds (in situation about being suitable for, being included in the pharmaceutical preparation), and therefore preferred these salt.

Because free form and its salt form of compound (comprise that those can be used as intermediate, the salt that for example is used for the intermediate of the purifying of described compound or its salt or evaluation) close relation between, in context, at any time relate to " compound " (also comprising parent material and " intermediate "), especially during the compound of formula I, if suitable and easily in the situation and do not have an other clearly description, then it is appreciated that the mixture that also relates to its one or more salt or free cpds and its one or more salt, it also comprises any solvate of formula I compound separately, metabolic precursor thereof such as ester or acid amides, the salt of perhaps any or multiple these materials.Can also obtain different crystalline forms, and the present invention also comprises these crystalline forms.

Use in the situation of plural number in compound, salt, pharmaceutical preparation, disease, illness etc., also mean the compound that relates to odd number, salt, pharmaceutical preparation, disease etc., in the situation of using " a kind of ", its perhaps article or preferably finger " ".

In some cases, compound of the present invention can comprise and one or morely is arranged in the chiral centre of substituting group or shows other asymmetry (causing producing enantiomer) or can also exist with the form of more than one steric isomers, for example owing to have an above chiral centre or above asymmetric center or owing to exist and make ring that Z/E (or cis-trans) isomery (diastereomer) occurs or two key and exist with the form of more than one steric isomers.The present invention includes two or more such mixture of isomers, as the mixture of enantiomer, racemoid especially, and the enantiomer of isomer, the especially purifying of preferred purifying or the mixture of enantiomer enrichment.

The compound of formula I has valuable pharmacological character and can be used for treatment to protein kinase, especially (especially " general description of the present invention " descends defined one or more protein kinases to protein tyrosine kinase above one or more, the most especially c-src kinases, VEGF-receptor kinase (for example KDR and Flt-1), RET-receptor kinase and/or Ephrin receptor kinase, EphB2 kinases for example, EphB4 kinases or associated kinase kinases) adjusting the disease of response is arranged, wherein regulate preferably being meant and suppressing and response is meant that the process of disease and/or its symptom are slowed down, stop or even counter-rotating and comprising fully or at least temporarily cure.Term " treatment " especially comprises prevention (comprising prophylactic treatment), for example in having observed the patient who shows sudden change that it is easy to maybe may to be easy to form disease or variation, perhaps preferably refer to described treatment of diseases processing (comprise without limitation take stopgap measures, healing, relief of symptoms, minimizing symptom, suppress disease or symptom, delay process, kinases is regulated and/or kinase inhibition), the especially any or multiple disease described below of described disease.

Term used herein " healing " preferably is meant in treatment and relates to effect in the active occurent incident of receptor tyrosine kinase (especially imbalance).Term " prevention " is meant that preferably prevention relates to the outbreak or the recurrence of the disease of receptor tyrosine kinase activity imbalance.

Term used herein " delay of process " refers in particular to treats disease early stage or early stage patient uses described active compound to being in, wherein said patient for example be diagnosed as be in corresponding disease early stage form or described patient be in a kind of situation, for example in the process of carrying out therapeutic treatment or be in, perhaps think and for example under the situation of not treating, can shift by may will forming in the situation of corresponding disease that mishap causes.

Animal is warm-blooded animal preferably, more preferably is Mammals.People's (it also drops in the generic term " animal " usually) especially has the patient that suffers from the disease risks that defines hereinafter or people (for example owing to some sudden changes or other characteristic ill risk being arranged).

Mention that at context term " application " (as verb or noun) is when (relating to the application of the compound or pharmaceutically acceptable salt thereof of formula I), suiting with at one's leisure, if not opposite explanation, (if not providing different expressions or instruction in context) then its comprise the embodiment (if having explanation) below any or multiple the present invention respectively: albumen (especially tyrosine) kinases is regulated (especially inhibition) and had application in the disease of response in treatment, being used for preparation treatment regulates (especially suppressing) to protein kinase the application of pharmaceutical composition of the disease of response is arranged, use the method for one or more formulas I compound in that protein kinase is regulated that (especially suppressing) have a response in treatment and/or the proliferative disease, be used for the treatment of the described pharmaceutical preparation that comprises one or more formulas I compound that protein kinase adjusting (especially suppressing) is had the disease of response, describedly protein kinase is regulated (especially suppressing) compound of one or more formulas I of the disease of response is arranged with being used for the treatment of.Treated and disease therefore formula I compound preferred " applications " is selected from described belowly regulates (especially inhibition) to albumen (especially tyrosine) kinases and has response (not only to refer to " dependence ", also refer to " support ", comprise that also disease wherein has the situation (be the activity support of protein kinase or even cause the disease performance) of response to the adjusting (especially suppressing) of protein kinase) disease, especially following proliferative disease.

In mentioning the situation of protein kinase, it is meant the protein kinase of any kind, especially one of defined these kinases under " general description of the present invention " above, more particularly serine/threonine and/or optimization protein Tyrosylprotein kinase, one or more tyrosine protein kinase most preferably, especially be selected from the c-src kinases, VEGF-receptor kinase (for example KDR and Flt-1), RET-receptor kinase and/or Ephrin receptor kinase, EphB2 kinases for example, EphB4 kinases or associated kinase kinases, comprise that any or multiple these kinase whose one or more changes or sudden change or equipotential form (for example make each proto-oncogene change into these forms of oncogene, as form activatory mutant, for example Bcr-Abl).Especially comprise unusual high expression level, form activatory or normal but in the form of giving in the stable condition hyperactivity hyperkinesia relatively and/or sudden change of other regulation mechanism of patient.

The purposes of The compounds of this invention in protein kinase the is regulated purposes of inhibitor (especially as) can prove with the pilot system of the protein kinase of preferably mentioning above following being used for:

In the description of model experiment property testing system below, following abbreviation has following implication: the DMSO=dimethyl sulfoxide (DMSO); The DTT=dithiothreitol (DTT); The EDTA=edetate; The MOI=infection multiplicity; PMSF=is right-the tolylsulfonyl fluorine; Tris=three (hydroxymethyl) aminomethane.If not explanation in addition, then " inhibitor " is meant the formula I compound of being tested.

Can following such effect that proves formula I compound as the kinase whose inhibitor of Ephrin B4 acceptor (EphB4):

Bac-to-Bac ^TMThe generation of (Invitrogen Life Technologies, Basel, Switzerland) GST-fusion expression vector:, increased in intact cell matter (cytoplasmatic) coding region of EphB-class respectively by the cDNA storehouse that derives from people's placenta or brain by PCR.Produce the recombinant baculovirus (SwissProt Database, registration number P54760) in the 566-987 amino acids zone of expressing human EphB4 acceptor.With the GST sequence clone to pFastBac1

In the carrier (Invitrogen LifeTechnologies, Basel, Switzerland) and with its pcr amplification.The cDNA of coding EphB4-receptor domain is cloned in 3 ' prime frame of GST sequence respectively in the FastBac1 carrier of this modification to produce pBac-to-Bac ^TMThe donor carrier.The single bacterium colony that derives from this conversion is cultivated, thereby obtained overnight culture, carry out the preparation of small-scale plasmid with it.The restriction enzyme analysis of plasmid DNA shows that some clones comprise and expects the inset of yardstick.By automatic sequencing, confirm in two strands, all to have inserted flank (flanking) the carrier sequence of about 50bp.

The preparation of virus: if not otherwise specified, then the scheme according to GIBCO prepares each kinase whose virus.Briefly, the transfer vector transfection that will comprise described kinase domain is coated onto on the selectivity agar plate in DH10Bac clone (GIBCO) and with it.Do not contain the bacterium colony that is inserted into the fusion sequence in the viral genome (entrained) and be blueness by this bacterium.Pick out one white colony and from bacterium, isolate viral DNA (rod granule) with the standard plasmid purification process.Then, according to this scheme, with Cellfectin reagent with Sf9 cell or Sf21 cell at 25cm ²Flask in use the viral DNA transfection.

The kinase whose purifying of GST-mark: the centrifugal cellular lysate loads on gsh-agarose column (Pharmacia) of a 2mL, with 10mL 25mM Tris-HCl, and pH 7.5,2mM EDTA, 1mM DTT, 200mM NaCl washing three times.Then, by using (each 1mL) 25mM Tris-HCl 10 times, pH 7.5,10mM reduced glutathion, 100mMNaCl, 1mM DTT, 10% glycerine come the albumen of wash-out GST-mark and it are stored under-70 ℃.

Protein kinase test: by measuring by [γ ³³P] ATP is blended into as the L-glutamic acid of substrate and tyrosine polymkeric substance (in poly-(Glu, Tyr)) ³³P comes existing or not existing the protein kinase activity under the inhibitor situation to analyze.The kinase assay of this use purifying GST-EphB (30ng) was carried out 15-30 minute with the final volume of 30 μ L at ambient temperature, in described volume, comprise 20mM Tris-HCl, pH 7.5,10mM MgCl ₂, 3-50mM MnCl ₂, 0.01mM Na ₃VO ₄, 1%DMSO, 1mM DTT, 3 μ g/mL poly-(Glu, Tyr) 4: 1 (Sigma; St.Louis, Mo., USA) and 2.0-3.0 μ M ATP (γ-[ ³³P]-ATP 0.1 μ Ci).Finish this test by adding 20 μ L 125mM EDTA.Subsequently, before 40 μ l reaction mixtures are transferred to methyl alcohol soaked 5 minutes the Immobilon-PVDF film (micropore, Bedford, MA, USA) on, water cleans, and uses 0.5%H then ₃PO ₄Soaked 5 minutes, it is placed on the vacuum manifold of the vacuum source with disconnection.After placing all samples, connect vacuum and with each hole with 200 μ l 0.5%H ₃PO ₄Wash.Take off these films and it is used 1.0%H on wobbler ₃PO ₄Wash 4 times, use washing with alcohol 1 time.It is dry at ambient temperature, be placed on the framework of Packard TopCount 96-hole, add the Microscint in 10 μ L/ holes ^TM(Packard) after, these films are counted.Carry out linear regression analysis by inhibition per-cent and calculate IC each test compound of duplicate, 4 concentration (being generally 0.01,0.1,1 and 10 μ M) ₅₀Value.A protein kinase activity unit is defined under 37 ℃, and the every mg albumen of per minute is from [γ ³³P] ATP shifts 1nmol to substrate protein ³³P ATP.The compound of formula I shows the EphB4 that is low to moderate 1nM and suppresses its IC ₅₀Value is preferably 0.001-10 μ M.

Perhaps, can be as described below the autophosphorylation effect of measurement EphB4 acceptor:

Use a kind of in vitro tests, with the A375 human melanoma cell of cell such as transfection (ATCC number: CRL-1619) confirm inhibition to the effect of EphB4 acceptor autophosphorylation, described cell permanent table intelligent EphB4 (SwissProt AccNo P54760), with its be inoculated into the perfect medium that is arranged in 6-porocyte culture plate (contain 10% peptide bovine serum=FCS) and with its under 37 ℃ at 5%CO ₂Be cultured to down and show about 90% fusion rate.Then, test-compound is diluted with substratum (do not contain FCS, contain 0.1% bovine serum albumin) and join in the described cell.(contrast comprises the substratum that does not contain test compounds).By adding 1 μ g/ml solubility ephrinB2-Fc (s-ephrinB2-Fc:R﹠amp; DBiosystems, CatNr 496-EB) and 0.1 μ M ortho-vanadate come the effect of inducing ligand inductive autophosphorylation.With it after cultivating 20 minutes again under 37 ℃, with cell with ice-cold PBS (salt solution of phosphate buffered) washed twice and with it immediately with the amount dissolving of every hole 200 μ l dissolving damping fluid.Then, this lysate is centrifugal to remove nucleus, with the protein concentration in commercial albumen test (PIERCE) the mensuration supernatant liquor.Then, use this lysate or if necessary immediately, it is stored under-20 ℃.

Carry out sandwich ELISA to measure the EphB4 phosphorylation: in order to catch the EphB4 albumen of phosphorylation, with the quantity in 100ng/ hole with ephrinB2-Fc (s-ephrinB2-Fc:R﹠amp; DBiosystems, CatNr 496-EB) be fixed on MaxiSorb (Nunc) elisa plate.Then, be positioned at the washing of this plate and with 3% and have polysorbas20 (polyoxyethylene (20) Arlacel-20, the TopBlock in the physiological saline of phosphate buffered ICI/Uniquema) (PBST)

(Juro, Cat.#TB232010) saturated remaining floating preteins combining site.Then, cellular lysate (every hole 100 μ g albumen) was at room temperature cultivated in these plates 1 hour.After these holes being washed 3 times, add with the anti-phosphotyrosine antibody of alkaline phosphatase bonded (PY20 alkaline phosphatase bonded: ZYMED, Cat Nr03-7722) and with it and cultivated again 1 hour with PBS.Once more these plates are washed, then with 10mM D-nitrophenyl phosphoric acid ester as the combining of substrate proof and quantitative anti-phosphotyrosine antibody and the phosphorylation acceptor that is hunted down, and after 0.5 hour-1 hour, its OD of measurement under 405nm.

Difference between positive control (stimulating with vanadate and s-ephrinB2-Fc) signal and negative control (stimulation) signal is equivalent to maximum EphB4 phosphorylation (=100%).Suppress the form of per-cent with maximum EphB4 phosphorylation and calculate the activity of being tried material, wherein induce half maximum material concentration that suppresses to be defined as IC ₅₀(the 50% inhibition dosage that suppresses).IC that can discoverable type I compound ₅₀Value is 0.0005 to 20 μ M, is preferably 0.0005 to 10 μ M.

The compound of formula I also can suppress other tyrosine protein kinase, c-Src kinases especially for example, it is animal, especially comprise people's cell mammalian cell growth regulating and transform in work.People such as Andrejauskas-Buchdunger, Cancer Res.52,5353-8 (1992) has described a kind of suitable test.Use this pilot system, the IC of the inhibition c-Src that the compound of formula I shows ₅₀Value is generally 0.005 to 10 μ M for for example 0.001 to 20 μ M.

Can following such activity that proves The compounds of this invention as KDR albumen-tyrosine kinase activity inhibitor: confirm inhibition to the effect of VEGF-inductive acceptor autophosphorylation with the Chinese hamster ovary celI of cell such as transfection, described cell permanent table intelligent VEGF-R2 acceptor (KDR), with its be inoculated into the perfect medium that is arranged in 6-porocyte culture plate (contain 10% peptide bovine serum=FCS) and with its under 37 ℃ at 5%CO ₂Be cultured to it down and show about 80% fusion rate.Then, test-compound is diluted with substratum (do not contain FCS, contain 0.1% bovine serum albumin) and join in the described cell.Contrast comprises the substratum that does not contain test compounds.It after 2 hours, is being added reorganization VEGF in cultivation under 37 ℃; The final concentration of VEGF is 20ng/ml.With it after cultivating 5 minutes again under 37 ℃, with cell with ice-cold PBS (salt solution of phosphate buffered) washed twice and with it immediately with the quantity dissolving of every hole 100 μ l dissolving damping fluid.Then, this lysate is centrifugal to remove nucleus, with the protein concentration in commercial albumen test (BIORAD) the mensuration supernatant liquor.Then, use this lysate or if necessary immediately, it is stored under-20 ℃.Use this scheme, the IC that the KDR that the alternative cpd of discoverable type I shows suppresses ₅₀Value is preferably compared the IC of c-Abl Tyrosylprotein kinase ₅₀Be worth at least 1.5 times of height, more preferably compare the IC of EphB4 Tyrosylprotein kinase ₅₀Be worth high 2 times.The IC of the compound of discoverable type I in this pilot system ₅₀Value is generally 0.001 to 20 μ M, more preferably is 0.005 to 10 μ M.

The compound of formula I also can suppress other protein kinase.

Can following such effect that proves The compounds of this invention as c-Abl albumen-tyrosine kinase activity inhibitor:

As people such as Geissler at Cancer Res.1992; Filter described in the 52:4492-4498 carries out the vitro enzyme test like that in conjunction with test in the 96-orifice plate, the modification below the described test of people such as Geissler has been carried out.As people such as Bhat at J.Biol.Chem.1997; The kinase domain of the His-mark of the such c-Abl of clone described in the 272:16170-16175 and it is expressed in baculovirus/Sf9 system.Operate in that the albumen (c-Abl kinases) to 37kD carries out purifying on the cobalt metal chelating column with a kind of two steps, with anion-exchange column it is carried out purifying then, yield is 1-2mg/L Sf9 cell people such as (, the bibliography of being quoted) Bhat.After Coomassie blue stain, SDS-PAGE identifies the kinase whose purity of this c-Abl＞90%.This test comprises (cumulative volumes of 30 μ L): c-Abl kinases (50ng), 20mM TrisHCl, pH 7.5,10mM MgCl ₂, 10 μ M Na ₃VO ₄, 1mM DTT and 0.06 μ Ci/ test [γ ³³P]-ATP (5 μ M ATP), use 30 μ g/mL to gather-Ala, Glu, Lys, Tyr-6: there is 1%DMSO in 2: 5: 1 (Poly-AEKY, Sigma P1152).By add that 10 μ L 250mMEDTA finish to react and 30 these reaction mixtures of μ L are transferred to before with methyl alcohol soaked 5 minutes the Immobilon-PVDF film (micropore, Bedford, MA, USA) on, water cleans, and uses 0.5%H then ₃PO ₄Soaked 5 minutes, it is placed on the vacuum manifold of the vacuum source with disconnection.After placing all samples, connect vacuum and with each hole with 200 μ l 0.5%H ₃PO ₄Wash.Take off these films and it is used 1.0%H on wobbler ₃PO ₄Wash 4 times, use washing with alcohol 1 time.It is dry at ambient temperature, be placed on the framework of Packard TopCount 96-hole, add the Microscint in 10 μ L/ holes ^TM(Packard) after, these films are counted.Use this system, formula I compound suppresses to show for example IC of 0.002 to 100 μ M to c-Abl ₅₀Value, its IC ₅₀Value is generally 0.002 to 5 μ M.

In addition, can also suppress b-raf (V599E) with the compound of formula I.Exist or do not exist under the situation of inhibitor, by measuring by [γ ³³P] ATP is blended among (His)-I κ B ³³P comes the B-Raf-V599E activity is analyzed.Test compound is dissolved among the DMSO (10mM) and with it is stored under-20 ℃.New preparation serial dilution thing and further it is diluted in DMSO with pure water, thus obtain to be arranged in the testing liquid of 3 times of concentration of 3%DMSO.The final volume of this test (30 μ l) comprises 10 μ l testing liquids (1%DMSO), 10 μ l test mixtures (20mMTris-HCl, pH 7.5,3mM MnCl ₂, 3mM MgCl ₂, 1nM DTT, 3 μ g/ml (His)-I κ B, 1%DMSO and 3.5 μ M ATP[γ ³³P]-ATP 0.1 μ Ci) and 10 μ l enzyme dilutions (600ng GST-B-Raf-V599E).Programme with at MultiPROBE Iix to moving the liquid step, carry out this step with the 96-well format on MultiPROBE IILx or the HamiltonSTAR automation.This test is carried out (seeing people such as C.Garcia-Echeverria, Cancer Cel.. as described in the document 5, 231-9 (2004)), finish test by adding 20 μ l 125mM EDTA.Followingly carry out catching of phosphorylated peptide by the filter combined techniques like that: 40 μ l reaction mixtures are transferred in advance with methyl alcohol soaked on 5 minutes the Immobilon-PVDF film, water cleans, and then it is used 0.5%H ₃PO ₄Soaked 5 minutes and it is placed on the vacuum manifold of the vacuum source with disconnection.After placing all samples, connect vacuum and with each hole with 200 μ l 0.5%H ₃PO ₄Wash.Take off free film and it is used 1.0%H on wobbler ₃PO ₄Wash 4 times, use washing with alcohol 1 time.It is dry at ambient temperature, be placed on the framework of Packard TopCount96-hole, add the Microscint in 10 μ L/ holes ^TMAfter, these films are counted.At last, these plates are sealed and in microtest plate scintillometer (TopCount NXT, TopCount NXT HTS), it is counted.In the situation of dodging the plate method, at first on based on the plastic plate of polystyrene, carry out kinase reaction, after 60 minutes, come termination reaction then by adding 20 μ l 125mM EDTA.For catch (60 minutes, RT), biotinylated substrate is transferred on the sudden strain of a muscle plate that scribbles nickel.With test board with PBS washing three times and it is at room temperature dry.These plates sealed and in microtest plate scintillometer (TopCount NXT, TopCount NXT HTS), it is counted thereafter.By down duplicate to four concentration (being generally 0.01,0.1,1 and 10 μ M) or with since 10 μ M then the inhibition per-cent of the compound of 8 a single point forms of dilution in 1: 3 carry out linear regression analysis and calculate IC ₅₀Value.The IC that the b-raf that formula I compound shows suppresses ₅₀Value is 0.05 to 50 μ M.

These results show that the compound of formula I has favourable affinity.

Also have some to be used to prove the active in vivo test of formula I compound anti-tumor in vivo.For example, for whether the compound of analysis mode I can suppress vasculogenesis in the body, with mouse somatomedin planting model its influence to angiogenesis factor such as VEGF, bFGF, S-1P, PDGF or the response of IGF-1 inductive vasculogenesis is analyzed: porous tetrafluoroethylene cell (volume is 0.5mL) is comprised heparin with 0.8%w/v, and (20 units/ml), the agar that comprises or do not comprise somatomedin (2 μ g/ml people VEGF) are filled, in its subcutaneous dorsal part flank that is implanted to the C57/C6 mouse.On the same day of implanting described mouse, with test compounds (for example 5,10,25,50 or 100mg/kg, p.o., once a day) or matrix is handled mouse and continue thereafter to handle 4 days.When processing finishes, mouse is killed and takes out described cell.Carefully take off in the vascular tissue of this cell growth on every side and to it and weigh, come blood content is assessed (Drabkins method by the content of hemoglobin of measuring this tissue; Sigma, Deisenhofen, Germany).Respond with quantitative vasculogenesis as the metric Tie-2 protein level of endothelial marker thing with specific ELISA mensuration.Shown that before these somatomedins have induced the increase of weight, blood content and the Tie-2 protein level of this tissue (histologic characteristics is for comprising inoblast and little blood vessel) of growth around the cell and (seen people such as Wood JM by neutralizing antibody (for example specific neutralize VEGF) this response capable of blocking in the mode of dose-dependently, Cancer Res.60 (8), 2178-2189, (2000); With people such as Schlaeppi, J.Cancer Res.Clin.Oncol.125,336-342, (1999)).Use this model, shown restraining effect under the concentration that the compound of formula I provides in the above.

On the preferred meaning of the present invention, it is that a kind of treatment individuality that is subjected to is to albumen (preferred tyrosine) kinases that protein kinase is regulated the disease that response is arranged, especially the illness that the adjusting of preferred kinase activity above (especially suppressing) responds in useful mode, in described treatment, can use the compound of formula I, described disease is one or more proliferative disease (a kind of diseases that depend on protein kinase activity (especially unsuitable activity)), comprise the hyper-proliferative illness, as the illness below one or more: leukemia, hyperplasia, fibrosis (especially pulmonary fibrosis, but also can be the fibrosis of other type, as renal fibrosis), vasculogenesis, psoriatic, proliferation of smooth muscle in atherosclerosis and the blood vessel is as the narrow or restenosis of postangioplasty.In addition, the compound of formula I also can be used for treating thrombosis and/or scleroderma.

The compound of preferred formula I is being treated (comprising prevention) proliferative disease (especially to albumen (preferred tyrosine) kinases, the adjusting of the preferred kinase activity of mentioning especially here (especially suppress) has the disease of response) in application, described disease is selected from tumour or Cancerous disease, especially be preferably optimum or especially malignant tumour or Cancerous disease, it more preferably is solid tumor, the cancer of the brain for example, kidney, liver cancer, adrenal carcinoma, bladder cancer, breast cancer, cancer of the stomach (the especially tumour of stomach), ovarian cancer, colorectal carcinoma, the rectum cancer, prostate cancer, carcinoma of the pancreas, lung cancer (for example minicell or maxicell lung cancer), carcinoma of vagina, thyroid carcinoma, sarcoma, glioblastoma, multiple myeloma or gastrointestinal cancer, especially colorectal carcinoma or colorectal adenomas, or the tumour of head and neck, the for example head and the squamous cell carcinoma of neck, comprise tumorigenesis, especially the tumorigenesis of epithelial character is for example in the situation of breast cancer; Epidermis hyperplasia (not being cancer), especially psoriatic; Hyperplasia of prostate; Or leukemia.

The compound of formula I or its application make and can cause the formation of tumor regression and/or prophylaxis of tumours metastatic tumor and the growth of metastatic tumor (also comprising micro metastasis).

Can also relate to some or especially one albumen (preferred tyrosine) kinases with the compounds for treating of formula I, the disease of immune system of especially mentioned preferred protein kinase; In addition, the compound of formula I also can be used for treatment and wherein relates to the central or peripheral nervous system disease that signal that at least a albumen (preferred tyrosine) kinases, especially mentioned preferred protein tyrosine kinase carry out transmits.

In chronic granulocytic leukemia (CML), the chromosome translocation of interaction balance has produced the BCR-ABL fusion gene in the hemopoietic stem cell (HSC).The Bcr-Abl fusion rotein that the latter encodes carcinogenic.Although the tight controlled protein tyrosine kinase (it plays basic role in the adjusting of cell proliferation, adhesion and apoptosis) of ABL coding, but BCR-ABL fusion gene code set becomes activatory kinases (producing a kind of phenotype that shows clonal proliferation out of control, reduces and the apoptosis response that mutagenesis stimulates is reduced to the adhesive capacity of bone marrow interstital thereby this kinases transforms HSC), thereby makes and accumulated how pernicious distortion gradually.The gained granulocyte can not form sophisticated lymphocyte and be released in the circulation, thereby has caused the not enough and susceptibility increase of mature cell.The ATP competitive inhibitor of having described Bcr-Abl (or suitable mutant form) suppresses kinases (for example P-3 kinases and STAT5), make it can not activate mitotic division and anti--apoptotic pathways, thereby cause the necrocytosis of BCR-ABL phenotype that therefore the method for effective treatment CML also is provided.Therefore, the present invention is particularly useful for treatment as the compound of the formula I of Bcr-Abl inhibitor and crosses expression diseases associated, especially leukemia with it, as leukemia, and for example CML or ALL.

Vasculogenesis is regarded as and grows to the absolute prerequisite that maximum diameter surpasses the tumour of about 1-2mm; When growing to this limit, must provide oxygen and nutrition to tumour cell by diffusion.Therefore, no matter its origin and cause are how, after reaching certain size, each growth of tumor all depends on vasculogenesis.The mechanism that plays an important role in the oncotherapy activity of angiogenesis inhibitor mainly contains three kinds: 1) suppress blood vessel, especially capillary vessel to avascular dormancy tumor growth, the result owing to reach balance between apoptosis and propagation, does not have tumour to grow only; 2), thereby stoped the migration of tumour cell owing to do not have blood to flow into and flow out tumour; With 3) inhibition of endothelial cell proliferation, thus avoided being usually located at the paracrine growth stimulation of the endotheliocyte of blood vessel inner layer to the surrounding tissue performance.

In view of it has the ability that suppresses KDR and especially Ephrin receptor kinase and possible other protein kinase and therefore has the ability of regulating vasculogenesis, therefore the compound of formula I is particularly useful for treatment and corresponding acceptor (preferred tyrosine) the unsuitable activity of kinases, especially crosses expression diseases associated or illness.In these diseases, the particularly important is (for example ischemic) retinopathy, (for example relevant) macular degeneration with the age, psoriatic, fat, hemangioblastoma, vascular tumor, inflammatory diseases such as rheumatoid or rheumatic inflammatory disease, especially sacroiliitis such as rheumatoid arthritis, or other chronic inflammatory illness such as chronic asthma, artery or post-transplantation atherosclerosis, endometriosis, and ND especially, for example so-called solid tumor (gi tract especially, pancreas, breast, stomach, uterine neck, bladder, kidney, prostate gland, ovary, uterine endometrium, lung, the cancer of brain, melanoma, kaposi's sarcoma, the squamous cell carcinoma of head and neck, malignant pleural mesothelioma, lymphoma or multiple myeloma) and liquid tumors (for example leukemia).

The compound of formula I is particularly useful for treating or prevents by continuing the disease that vasculogenesis triggers, as restenosis, for example, the restenosis of stent-induced; Crohn's disease; Hodgkin; Eye disease is as diabetic retinopathy and neovascular glaucoma; The kidney disease is as glomerulonephritis; Diabetic nephropathy; Inflammatory bowel; Malignant nephrosclerosis; Embolic microangiopathy syndrome; (for example chronic) transplant rejection and glomerulopathy; Fibrotic conditions is as liver cirrhosis; Mesangial cell-proliferative disease; Neural tissue injury; Be used to suppress balloon catheter handle the back blood vessel closed again, be used for the blood vessel prosthodontics or be used to keep use behind the mechanism of vessel open such as the support in insertion, as immunosuppressor, be used to help the scar-free wound healing and be used for the treatment of senile plaque and contact dermatitis.

The present invention preferably relates to the compound of formula I or its pharmaceutically useful salt in for example application in the leukemia of treatment solid tumor described herein and/or liquid tumors described herein.

Because it regulates the character of protein kinase such as Eph receptor kinase, the compound of formula I also can be used for stimulating or promoting neurotization (neuron regeneration; Neurotization), perhaps suppresses or reverse neurodegeneration (neuronal degeneration as axonal regeneration; Neurodegeneration).Other aspects of the present invention have been represented in these application.

Therefore, the compound of formula I also can be used for treating situation, disease or the illness that the adjusting of protein kinase such as Eph receptor kinase is had response, wherein, wishes to stimulate or promote neurotization (neuron regeneration; Neurotization) as axonal regeneration or inhibition or reverse neurodegeneration (neuronal degeneration; Neurodegeneration), for example, it can be used for treating Spinal injury, hypoxemia situation, traumatic brain injury, infarct, apoplexy, multiple sclerosis or other neurodegeneration situation, disease or illness.These are used and methods of treatment has been represented other aspects of the present invention.

The preparation method

The compound of formula I is that the method with other compound of preparation well known in the prior art is prepared similarly, and for the novel cpd of formula I, this method also is new as similar approach, and its preparation method is preferably

A) incite somebody to action wherein R1, X ₁, X ₂, X ₃And X ₄Carboxylic acid suc as formula the defined formula of the compound of I (II)

Or the compound of its reactive derivatives and formula (III) carries out condensation

Wherein R2, R3, D, Y ₁, Y ₂Define suc as formula the compound of I with Ra, perhaps

B) for R1 wherein be not substituted or substituted alkyl, be not substituted or substituted aryl, be not substituted or substituted cycloalkyl be not substituted or the formula I compound of substituted heterocyclic radical synthetic for, wherein R1 is not substituted or substituted alkyl, is not substituted or substituted aryl (preferably), is not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical (separately by the combination of C-atom) and A are hydroxyl or lower alkoxy or B (A) ₂Be assorted two ring [3.3.1] nonanes of 9-boron or-B (CHCH ₃CH (CH ₃) ₂) ₂The boric acid of formula (IV)

R1-B(A) ₂(IV)，

With wherein R1, R2, R3, X ₁, X ₂, X ₃, X ₄, Y ₁, Y ₂, D and Ra define suc as formula the compound of I and Hal is a halogen, especially chlorine, iodine or preferably bromine or (perfluorination C ₁-C ₄-alkyl)-compound of the formula V of sulfonyloxy reacts

And if necessary, the compound of formula I is changed into another kind of different formula I compound, the salt of the formula I compound that obtains is changed into free cpds or another kind of different salt, the free formula I compound that obtains is changed into its salt and/or the isomer mixture of the formula I compound that obtains is separated into one isomer.

The condensation reaction of carrying out with carboxylic acid or its reactive derivatives of formula X or XIV a) is preferably used reactive carboxylic acid derivatives (it uses like this) respectively; for example with reactive carboxylic acid derivatives, active ester or the carboxylic acid halides of symmetric anhydride or mixed acid anhydride form; for example chloride of acid carries out; for example exist tertiary nitrogen alkali for example to carry out under the situation of three low-grade alkylamines or pyridine; perhaps it can carry out in position, for example can carry out condensation existing original position to form under the situation of reagent of reactive ester.This reaction for example can by with described carboxylic acid and corresponding amine solvent in The suitable solvent, halon for example, as methylene dichloride, N, dinethylformamide, N, the N-N,N-DIMETHYLACETAMIDE, the N-N-methyl-2-2-pyrrolidone N-, also add for example triethylamine of suitable alkali in the mixture of methylene dichloride or two or more these kind solvents, diisopropyl ethyl amine (DIEA) or N-methylmorpholine carry out, if and form the reactive derivatives of the acid of formula II in position, then also be added in the suitable coupler of preferred reactive derivatives that original position forms the carboxylic acid of formula III, for example dicyclohexylcarbodiimide/I-hydroxybenzotriazole (DCC/HOBT); Two (2-oxo-3-oxazolidinyl) phosphonyl chloride (BOPCl); O-(1,2-dihydro-2-oxo--1-pyridyl)-N, N, N ', N '-tetramethyl-urea a tetrafluoro borate (TPTU); O-benzotriazole-1-yl)-and N, N, N ', N '-tetramethyl-urea a tetrafluoro borate (TBTU); (benzotriazole-1-oxygen base)-tripyrrole alkane-1-base phosphorus-hexafluorophosphate (PyBOP), 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride/hydroxybenzotriazole or/1-hydroxyl-7-azepine benzotriazole (EDC/HOBT or EDC/HOAt) or HOAt (separately or with (1-chloro-2-methyl-propenyl)-dimethyl amine coupling).For some other available couplers, referring to for example Klauser; Bodansky, Synthesis1972,453-463.Preferably this reaction mixture is being made an appointment with-20 to 50 ℃, especially 0 ℃ to 30 ℃ temperature for example stirs under the room temperature.

B) reaction is carried out under Suzuki coupling conditioned disjunction equal conditions, for example has palladium catalyst, as Pd (PPh ₃) ₄(wherein Ph is a phenyl) or Pd (dppf) Cl ₂(wherein dppf is 1,1 '-two (diphenylphosphino) ferrocene) and alkali such as alkaline carbonate for example under the situation of yellow soda ash, alkali metal alcoholate such as sodium ethylate, tertiary nitrogen alkali such as triethylamine or alkali metal phosphate such as potassiumphosphate, there is or do not exist for example ether of suitable solvent, as tetrahydrofuran (THF) or 1,4-dioxan, hydrocarbon, for example under the situation of toluene and/or water, in the temperature that for example raises, for example 30 ℃ are carried out in closed pressure-resistant reaction vessel to reflux temperature or higher temperature.

Optional reaction and conversion

The compound of formula I can be changed into the compound of another kind of formula I, for example above-mentioned reaction a) or b) in one of transform after finishing.

For example, R1 is that halo-aryl such as bromo-aryl are for example in the compound of the formula I of bromophenyl therein, halogen can be replaced by nitrogen-atoms bonded substituting group, for example replaced by morpholine-4-base, it can be by for example uncle-butanols potassium and suitable coupling catalyzer for example for example react in ether such as the tetrahydrofuran (THF) at The suitable solvent or solvent mixture under the situation of chlorination 2-(dimethyl-amino-)-2-xenyl-palladium (II) two norcamphyl phosphine complex bodys and carry out having highly basic such as alkali metal alcoholate with corresponding primary amines or secondary amine such as morpholine, preferably react at elevated temperatures, for example to reflux temperature, react at 30 ℃.

Can provide another example that formula I compound transforms, wherein in substituted aryl R1, exist nitro substituent-this nitro substituent can be reduced into corresponding amino substituting group, for example reduce with catalytic hydrogenation, for example can exist under the situation of Raney nickel at The suitable solvent or solvent mixture alcohol for example, as reducing in methyl alcohol or the ethanol, temperature of reaction for example is 0 to 50 ℃.

Can by with corresponding haloalkane for example methyl iodide react the amino substituting group in the formula I compound (especially as substituent amino of the aryl R1 among the formula I) changed into single-, two-or three-alkylating amino (being quaternary ammonium in the later case) substituting group; this reaction preferably under the situation that has tertiary nitrogen alkali such as triethylamine at The suitable solvent or solvent mixture N for example; N-two-(low alkyl group)-low-grade alkane acidyl acid amides; as N; carry out in the dinethylformamide, temperature of reaction is preferably 20 to 80 ℃.

R1 is replaced by iodine or bromine and may also have in the formula I compound of aryl (for example R1 is a 4-iodo-3-trifluoromethyl) of one or more other substituting groups such as trifluoro therein, bromine or iodine can be substituted or unsubstituted aryl, substitute as the 4-cyano-phenyl, can there be catalyzer in it by the aryl boric acid with corresponding replacement or unsubstituted formula IV, especially PdCl ₂(dppf) react under the situation and carry out,

Ar-B(OH) ₂(IV)，

Wherein Ar is not substituted or substituted aryl, in this reaction, also preferably there are alkali such as alkaline carbonate, yellow soda ash for example, this is reflected at The suitable solvent or solvent mixture, for example in the toluene the temperature that for example raises for example 30 ℃ to (preferably) reflux temperature, carry out.

Salt with formula I compound of at least one salt forming group can be prepared in a manner known way.For example, can by with metallic compound as an alkali metal salt of suitable organic carboxyl acid for example 2 ethyl hexanoic acid sodium salt, with organic alkali metal or alkaline earth metal compound, as corresponding oxyhydroxide, carbonate or supercarbonate, handle the salt that the formula I compound with acidic-group forms the formula I compound with acidic-group as sodium hydroxide or potassium hydroxide, yellow soda ash or salt of wormwood, corresponding calcium cpd or ammonia or suitable organic amine, preferably use stoichiometry or only a small amount of excessive salt-forming reagent.Can obtain the acid salt of formula I compound with usual manner, for example can obtain by compound with sour or suitable anionresin agent treated formula I.Can form and comprise for example inner salt of the formula I compound of free carboxy and free amine group of acid and alkaline salt forming group, for example can be by salt such as acid salt being neutralized to iso-electric point or forming inner salt by handling with ion-exchanger with carboxylic acid.

Can the salt of formula I compound be changed into free cpds with usual manner; For example can be by coming metal with suitable acid treatment or ammonium salt transforms and can transform acid salt by handling with suitable alkaline reagents.In both of these case, can use suitable ion-exchanger.

Can be in a manner known way, with three-dimensional heterogeneous mixture, for example the mixture separation of diastereomer becomes its corresponding isomer by suitable separation method.For example can non-enantiomer mixture be separated into its each diastereomer by fractional crystallization, chromatography, solvent distribution and similar operations.This separation can be carried out on the level of a kind of initial compounds or formula I compound itself.Can by form chromatography that diastereoisomeric salt for example forms salt or the chromatogram substrate by having chiral ligand with use by the chiral acid with enantiomeric pure for example HPLC separate enantiomer.

Can carry out aftertreatment and/or purifying to intermediate and end product according to standard method, for example can wait and carry out aftertreatment and/or purifying with chromatography, apportion design, (weight) crystallization.

Parent material

Parent material be in the prior art known, can commercial obtain or can be prepared according to method well known in the prior art.If do not mention especially; then can in suitable step, introduce and remove blocking group and undesirable reaction take place in the respective reaction step to avoid functional group; the method of introducing and removing blocking group has been described in context, for example can be referring to described reference under " general reaction conditions ".Those skilled in the art can determine easily that blocking group is whether useful or not need and need any blocking group.

In parent material, use R1, R2, R3, X ₁, X ₂, X ₃, X ₄, Y ₁, Y ₂, Ra, D, R6, A, R4, Z ₁, Z ₂, Z ₃, R ₅In the situation of n, if do not specify that in context then these symbols preferably have the given implication of formula I compound.

Parent material for example can preferably followingly be prepared like that:

For example; wherein R1 is not substituted or substituted alkyl, is not substituted or substituted aryl, is not substituted or substituted cycloalkyl or be not substituted or the parent material of the formula II of substituted heterocyclic radical (and by ring carbon atom carry out in conjunction with) can be prepared by the compound (protecting on carboxyl if necessary) of formula VI

Wherein Hal is a halogen, especially chlorine, iodine or preferably bromine or (perfluorination C ₁-C ₄-alkyl)-sulfonyloxy, with itself and top reaction b) down the compound of defined formula IV react b in the above) react under the described preferred reaction conditions, removing then no longer needs or undesirable blocking group.

Wherein Ra is that the compound of the part of the defined formula IA of compound of formula I and the formula III that D is NH for example can following such preparation:

Wherein D is NH, Y ₂Be C and Ra be formula IA as defined above part (wherein A is that the compound of the formula III of C (=O)-N (R4)) can be preferably by carboxylic acid or its reactive derivatives with formula VII,

Y wherein ₂Be C, obtain with the amine condensation of formula VIII

Thereby obtain wherein Y ₂It is the compound of the formula IX of C

Then, can obtaining wherein, D is the compound of the formula III of NH, the compound reduction of the formula IX that is about to as before or obtains with any other method, for example it is reduced by catalytic hydrogenation, for example exist under the situation of Raney nickel at The suitable solvent or solvent mixture, for example pure, as for example under 0 to 50 ℃ the situation it being reduced in methyl alcohol or the ethanol, thereby obtain A among the Ra wherein and be C (=O)-N (R4) and R6 are the respective compound of the formula III of hydrogen, thereafter, if necessary, carrying out alkylation by the suitable halogenide with formula X under conventional alkylation conditions converts it into and is not substituted or substituted alkyl R6

R6-Hal(X)，

Wherein R6 is not substituted or substituted alkyl and Hal are halogens, especially bromine or iodine.

Perhaps, but wherein D is that the respective compound of the formula III of O or S can be by will similarly to top formula VII wherein not having nitro but have the amine condensation of the carboxylic acid cpd of hydroxyl (preferred protected hydroxyl) or sulfydryl (preferred protected sulfydryl) or its reactive derivatives and aforesaid formula VIII and remove blocking group and obtain.

Wherein D is NH, O or S, Y ₂Be that C and Ra are that (wherein A is that the compound of the formula III of N (R4)-C (=O)) can be by carboxylic acid or its reactive derivatives with formula XI for part as top defined formula IA

Obtain with the aminocompound condensation of formula XII,

Y wherein ₂Be C, D ^*Be (preferably protected) amino, (preferably protected) hydroxyl, (preferably protected) sulfydryl or nitro, thus the respective compound of the formula of acquisition XIII,

D wherein ^*Be (preferably protected) amino, (preferably protecting) hydroxyl, (preferably protecting) sulfydryl or nitro and Y ₂Be C,

Then, can convert it into the compound of corresponding formula III, that is, if D ^*Be the words of nitro; nitroreduction is become amino; for example reduce by catalytic hydrogenation; for example exist under the situation of Raney nickel at The suitable solvent or solvent mixture; for example pure; as for example reducing under 0 to 50 ℃ the temperature in methyl alcohol or the ethanol; thereby obtain wherein in Ra A and be N (R4)-C (=O) and R6 be hydrogen (thereafter; if necessary; can be undertaken by suitable halogenide with formula X as defined above alkylation convert it into be not substituted or substituted alkyl R6) corresponding formula III compound; perhaps, remove blocking group.

Preferably a) described reaction conditions is similar down to top method for the reactive derivatives of the carboxylic acid of formula VII or XI and the reaction conditions of condensation.

Other parent material, and as these materials that the parent material of top intermediate is mentioned be in the prior art known, can commercial obtain and/or can be prepared according to standard operation, for example can be prepared with the method described in the embodiment or with its similar methods.

General treatment condition

Following narration is applicable to described all methods of context usually, the simultaneously preferred specific reaction conditions of mentioning of context:

In the mentioned any reaction of context; in the situation that suits or need; mention even without specific, also can protect the functional group that does not need to participate in given reaction, and can be introduced into and/or remove in suitable or required stage with blocking group.Therefore, do not have specific mentioning in protection and/or the de-protected reaction in this manual, comprise the reaction of using blocking group when needed yet.

In the scope of this paper, unless stated otherwise, otherwise only not that the group of removing easily of the integral part of required specific formula I end product is called as " blocking group ".At for example canonical reference works, as J.F.W.McOmie, " blocking group in the organic chemistry (Protective Groups in OrganicChemistry) ", Plenum Press, London and New York 1973, T.W.Greene and P.G.M.Wuts, " blocking group in the organic synthesis (Protective Groups in Organic Synthesis) ", the third edition, Wiley, New York 1999, " peptide class (The Peptides) "; The 3rd volume (chief editor: E.Gross and J.Meienhofer), Academic Press, London and New York 1981, " Methoden derorganischen Chemie " (organic chemistry method), Houben Weyl, the 4th edition, the 15/1st volume, Georg Thieme Verlag, Stuttgart 1974, H.-D.Jakubke and H.Jesch-keit, "

Peptide; Proteine " (amino acid; peptide class; protein); Verlag Chemie; Weinheim; Deerfield Beach; with Basel 1982 and Jochen Lehmann; " Chemieder Kohlenhydrate:Monosaccharide und Derivate " (carbohydrate chemistry: monose and derivative), Georg Thieme Verlag, among the Stuttgart 1974 to the protection, the blocking group itself that functional group are carried out with blocking group and be applicable to that the reaction of removing described protecting group is described.The characteristic of blocking group is that it can easily be removed (promptly undesirable secondary reactions can not take place), for example can be removed or can be removed (for example being removed by enzymatic lysis) under physiological condition by solvolysis, reduction, photodissociation.

All above-mentioned treatment steps can carry out under known reaction conditions own; under the condition of preferably specifically mentioning in the above; do not exist or have solvent or thinner usually; be under the situation of the solvent of inertia and solubilized agents useful for same or thinner preferably to agents useful for same; there is not or exists catalyzer; condensing agent or neutralizing agent for example ion-exchanger such as cationite for example carry out under the situation of H+ type exchanger; character according to said reaction and/or reactant; reducing; normal or raise temperature; for example about-100 ℃ to about 190 ℃; preferred-80 ℃ to about 150 ℃ approximately; for example-80 to-60 ℃; room temperature;-20 to 40 ℃ or under reflux temperature; under atmospheric pressure or in sealed vessel; under suitable situation, depress and/or, for example carry out under argon gas or the nitrogen atmosphere adding at inert atmosphere.

Unless in the description of method, specify, can by the solvent of wherein selecting these solvents that are applicable to any specific reaction comprise solvent that those are specifically mentioned or, for example, water; The ester class, as low alkyl group-lower alkanoic acid ester, ethyl acetate for example; Ethers, as aliphatic ethers, for example ether, or cyclic ethers class, for example tetrahydrofuran (THF) or dioxan; The liquid aromatic hydrocarbons class is as benzene or toluene; Alcohols is as methyl alcohol, ethanol or 1-or 2-propyl alcohol; Nitrile is as acetonitrile; Halogenated hydrocarbon, for example methylene dichloride or chloroform; Amides is as dimethyl formamide or N,N-DIMETHYLACETAMIDE; Bases, as heterocyclic nitrogenous bases, for example pyridine or N-methylpyrrolidin-2-ketone; Carboxylic acid anhydride is as lower alkane acid anhydrides, for example diacetyl oxide; Ring-type, straight or branched hydro carbons are as the mixture of hexanaphthene, hexane or iso-pentane or these solvents, for example aqueous solution.In aftertreatment, for example also can use such solvent mixture in the aftertreatment of being undertaken by chromatography or distribution.

The invention still further relates to compound that any stage of wherein being used in the said method obtains with intermediate forms as the go forward side by side surplus treatment step in Xingqi or wherein forming parent material or parent material under the said reaction conditions with for example protected form of its derivative form or salt form is used or make the compound that obtains with the inventive method and on the spot to the method for its these forms of further handling under said treatment condition of parent material.In the method for the invention, preferably use these parent materials that produce preferred formula I compound.The invention still further relates to new intermediate and/or parent material.The same or analogous reaction conditions of these reaction conditionss described in the preferred especially and embodiment.

The preferred embodiments of the invention

Embodiment preferred and in front with the embodiment of following more general range in, and in claims, can replace any one or a plurality of or all generality to explain with the top and following corresponding more specifically definition that provides, thereby obtain the preferred embodiment of the present invention.

In a preferred embodiment, the present invention relates to the formula I compound of following free form or salt form, wherein

R1 is hydrogen, halogen, be not substituted or substituted alkyl, be not substituted or substituted aryl, be not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical;

R2 and R3 are hydrogen, halogen, C independently of one another ₁-C ₄Alkyl, trifluoromethyl, C ₁-C ₄Alkoxyl group or cyano group;

X ₁, X ₂, X ₃And X ₄Be CH;

D is N (R6), and wherein R6 is acyl group or is not substituted or substituted alkyl;

Y ₁Be HC=CH;

Y ₂Be C;

If R1 is unsubstituted alkyl, substituted alkyl with 5 or more a plurality of carbon atoms, be not substituted or substituted aryl, be not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical, then Ra is a hydrogen; Perhaps, if R1 is hydrogen, halogen or C ₁-C ₄-alkyl, then Ra is the part of the formula IA that provides previously, wherein

Dotted line represents and formula I molecule rest part bonded key,

A be C (=O)-N (R4) or N (R4)-C (=O), wherein R4 is hydrogen or is not substituted or substituted alkyl,

Z ₁Be CH=CH,

Z ₂Be N or CH,

Z ₃Be CH,

R ₅Be substituting group independently of one another, and

N is 0,1 or 2.

In another embodiment, the present invention relates to the formula I compound of following free form or salt form, wherein

R1 is hydrogen, halogen or C ₁-C ₄-alkyl;

R2 and R3 are hydrogen, halogen, C independently of one another ₁-C ₄-alkyl, trifluoromethyl, C ₁-C ₄-alkoxyl group or cyano group,

X ₁, X ₂, X ₃And X ₄Be that CH or wherein maximum two can be N;

D is N (R6) (preferably), O or S,

Wherein R6 is hydrogen, acyl group or is not substituted or substituted alkyl,

Y ₁Be O, S, NH, CH ₂, N=CH, CH=N or CH=CH;

Y ₂Be C;

Ra is the part of formula IA

Wherein dotted line is represented and the rest part of formula I molecule (with shown in the wavy line) bonded key,

A be C (=O)-N (R4) or N (R4)-C (=O),

Wherein R4 is hydrogen or is not substituted or substituted alkyl,

Z ₁Be O, S, NH, CH ₂, CH=N, N=CH or CH=CH,

Z ₂Be nitrogen or CH,

Z ₃Be CH or N,

Each R that occurs ₅Be a kind of substituting group independently of one another, and

N is 0,1 or 2.

In another preferred embodiment, the present invention relates to the formula I compound of following free form or salt form, wherein

R1 is unsubstituted alkyl or substituted alkyl with 5 or more a plurality of carbon atoms, be not substituted or substituted aryl, be not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical;

R2 and R3 are hydrogen, halogen, C independently of one another ₁-C ₄-alkyl, trifluoromethyl, C ₁-C ₄-alkoxyl group or cyano group,

X ₁, X ₂, X ₃And X ₄Be that CH or wherein maximum two can be N;

D is N (R6) (preferably), O or S,

Wherein R6 is hydrogen, acyl group or is not substituted or substituted alkyl,

Y ₁Be O, S, NH, CH ₂, N=CH, CH=N or CH=CH;

Y ₂Be C or N;

Ra do not exist or, if Y ₂Be C, Ra is a hydrogen.

The invention still further relates to pharmaceutical preparation, compound is regulated (especially suppressing) and is had the application in the disease of response or be used for the preparation treatment and protein kinase regulated (especially suppressing) application of pharmaceutical preparation of the disease of response is arranged to protein kinase in treatment, a kind of methods of treatment (it comprises to the individuality of such treatment of needs (animal or preferred people) has the amount of the disease of response to use compound or its pharmaceutically useful salt of formula I with the adjusting (especially suppressing) that is enough to treat to protein kinase (especially protein tyrosine kinase)), and the method for the compound or its salt of preparation formula I; In various situations, the compound of formula I or its (pharmaceutically useful) salt preferably have the compound of the described preferred feature of context.

The present invention also preferably relates to given embodiment in the claim (especially independent claim).Therefore, all claims here all are introduced into as a reference.

The present invention especially relates to compound or its pharmaceutically useful salt of formula I given among the embodiment or it is according to the method for mentioning among application of the present invention and the embodiment and new parent material and intermediate.

Pharmaceutical composition

The invention still further relates to disease or illness, preferred illness especially recited above or the method for disease, the compound that is used for described application and pharmaceutical preparation and preparation thereof that pharmaceutical composition, its application in therapeutic treatment (broadly also comprising preventative processing of the present invention) or the treatment of the compound that comprises formula I (preferred new compound) depend on unsuitable albumen (especially tyrosine) kinase activity, in particular for the pharmaceutical preparation of described application.In the situation of formula I compound, more generally use pharmaceutical preparation.

The acceptable compound of pharmacology of the present invention for example can be present in and comprise that significant quantity is inorganic or organic as the compound or pharmaceutically acceptable salt thereof of the formula I of activeconstituents and one or more, in the pharmaceutical composition of pharmaceutically acceptable carrier (carrier substance) of solid or liquid form, perhaps for example can be used for preparing such pharmaceutical composition.

The invention still further relates to and be suitable for delivering medicine to warm-blooded animal, especially the people (perhaps derives from warm-blooded animal, especially people's cell or clone, lymphocyte for example), the inhibition of kinase activity has the pharmaceutical composition of the disease of response to albumen (especially tyrosine) to be used for treatment (also comprising prevention aspect the generalized of the present invention), it comprises compound or its pharmaceutically useful salt and at least a pharmaceutically useful carrier of a certain amount of formula I, wherein, the amount of said compound preferably is effective for said inhibition.

Pharmaceutical composition of the present invention is to be used in the intestines as nose, rectum or oral or parenteral such as intramuscular or intravenous administration in warm-blooded animal (especially people's) pharmaceutical composition the pharmaceutically useful carrier that it only comprises the said pharmacologically active principles of effective dose or also comprises q.s.The dosage of said activeconstituents depends on kind, body weight, age and the individual instances of warm-blooded animal, individual pharmacokinetic data available, the disease and the administering mode of being treated.

The invention still further relates to a kind of treatment and the method for the disease of response is arranged depending on albumen (especially tyrosine) inhibition of kinase whose unsuitable active disease; It comprises especially gives the warm-blooded animal that needs such treatment owing to one of described disease, and for example the people uses formula I compound or its pharmaceutically useful salt that prevents or especially treat significant quantity.

Delivered medicine to warm-blooded animal, for example the dosage of the people's of the about 70kg of body weight formula I compound or pharmaceutically acceptable salt thereof is preferably about 3mg to about 10g, more preferably be that about 10mg is to about 1.5g, be most preferably about 100mg to about 1000mg/ people/sky, it preferably is divided into 1-3 single dose, and said single dose for example size can be identical.The dosage that children accepted is generally half of adult's dosage.

Said pharmaceutical composition comprises about 1% to about 95%, preferred about 20% to about 90% activeconstituents.Pharmaceutical composition of the present invention for example can be unit dosage form, for example can be ampulla, bottle, suppository, dragee, tablet or capsular form.

Pharmaceutical composition of the present invention can be prepared with known mode itself, for example can be prepared by dissolving, lyophilize, mixing, granulation or the forming method of routine.

The preferred solution that uses activeconstituents, and can use its suspension, and especially isoosmotic aqueous solution or suspension, for example only comprise activeconstituents or comprise activeconstituents and the situation of the freeze-dried composition of carrier such as N.F,USP MANNITOL in, can prepare such solution or suspension before use.Said pharmaceutical composition can be sterilized and/or can be comprised vehicle, for example sanitas, stablizer, wetting agent and/or emulsifying agent, solubilizing agent, be used to regulate the salt and/or the buffer reagent of osmotic pressure, and can be prepared with known mode itself, for example can be prepared by routine dissolving or freeze-drying method.Said solution or suspension can comprise the material that increases viscosity, as Xylo-Mucine, carboxymethyl cellulose, dextran, polyvinylpyrrolidone or gelatin.

The suspension that is arranged in oil comprises the vegetables oil that is usually used in injecting purpose as oily components, synthetic or semi-synthetic oils.Especially comprising of can mentioning has 8-22, especially the longer chain fatty acid as acid constituents of 12-22 carbon atom, for example lauric acid, tridecanoic acid, tetradecanoic acid, pentadecylic acid, palmitinic acid, margaric acid, stearic acid, eicosanoic acid, docosoic acid or corresponding unsaturated acid, for example oleic acid, elaidic acid, erucic acid, brassidic acid or linoleic liquid aliphatic acid esters, if necessary, also can add oxidation inhibitor, for example vitamin-E, β-Hu Luobusu or 3,5-two-tert-butyl-4-hydroxytoluene.The alkoxide component of these fatty acid esters have maximum 6 carbon atoms and be single-or many-hydroxyl for example single-, two-or three-hydroxyl alcohol, for example methyl alcohol, ethanol, propyl alcohol, butanols or amylalcohol or its isomer, still two pure and mild glycerine especially.Therefore, the example of the fatty acid ester that can mention is as follows: ethyl oleate, Isopropyl myristate, Wickenol 111, " Labrafil M 2375 " (polyoxyethylene triolein, Gattefoss é, Paris), " Miglyol 812 " (triglyceride level of saturated fatty acid with chain length of C8 to C12, H ü ls AG, Germany), but vegetables oil especially, as Oleum Gossypii semen, Prunus amygdalus oil, sweet oil, Viscotrol C, sesame oil, soya-bean oil and peanut oil.

Said injection or transfusion composition are prepared under aseptic condition in a usual manner; Under aseptic condition, said composition is incorporated in ampoule or the bottle also with this container sealing too.

Being used for pharmaceutical composition for oral administration can be by admixed together with activeconstituents and solid carrier, if necessary, with the granulating mixture of gained and if desired or essential, after adding suitable vehicle, this mixture is processed into tablet, dragee nuclear or capsule and obtains.It can also be blended into can be so that in the plasticity carrier of activeconstituents with determined amount diffusion or release.

Suitable carrier has weighting agent usually, as carbohydrate, lactose for example, sucrose, N.F,USP MANNITOL or sorbyl alcohol, cellulose preparation and/or calcium phosphate, for example tricalcium phosphate or secondary calcium phosphate, and tackiness agent, as starch paste, for example use corn, wheat, the starch paste of paddy rice or yam starch, gelatin, tragacanth gum, methylcellulose gum, HPMC, Xylo-Mucine and/or polyvinylpyrrolidone, and/or if necessary, disintegrating agent, as above-mentioned starchy material, and/or carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, alginic acid or its salt are as sodiun alginate.Vehicle is flowing regulator and lubricant especially, and for example silicic acid, talcum powder, stearic acid or its salt are as Magnesium Stearate or calcium stearate and/or polyoxyethylene glycol.For dragee nuclear provides suitable, optional is the dressing of enteric, for this reason, can adopt the dense sugar soln that can comprise gum arabic, talcum powder, polyvinylpyrrolidone, polyoxyethylene glycol and/or titanium dioxide, perhaps be arranged in the dressing solution of suitable organic solvent, perhaps, in order to prepare enteric coating, can adopt the solution of suitable cellulose preparation such as phthalic acid ethyl cellulose or phthalic acid HPMC.Capsule has the capsule of the dry-packing that is made by gelatin and the soft capsule of the sealing that made by gelatin and softening agent such as glycerine or sorbyl alcohol.The capsule of dry-packing can comprise the activeconstituents of particle form, said particle form for example can have weighting agent such as lactose, tackiness agent such as starchy material and/or glidant such as talcum powder or Magnesium Stearate, and can also have stablizer if necessary.In the situation of soft capsule, preferably with the dissolving of said activeconstituents or be suspended in suitable oiliness vehicle, in fatty oil, paraffin oil or liquid macrogol, can also be to wherein adding stablizer and/or antiseptic-germicide.Can in tablet or dragee coatings or capsule shell, add dyestuff or pigment, for example for identifying purpose or show the various dose of activeconstituents.

Also can be advantageously with compound and other biologic activity agent of formula I, preferably other antiproliferative is united use.Described antiproliferative comprises aromatase inhibitor without limitation; Antiestrogen; The topoisomerase I inhibitor; The topoisomerase II inhibitor; The microtubule activator; Alkylating agent; Histone deacetylase inhibitors; The compound of inducing cell atomization; Cyclooxygenase inhibitors; The MMP inhibitor; The mTOR inhibitor; Antineoplastic metabolic antagonist; Platinic compound; Target/reduction albumen or the active compound of lipid kinase and other anti-angiogenic compounds; The compound of target, reduction or arrestin or lipid phosphatase activity; Gonadorelin (gonadorelin) agonist; Antiandrogen; The methionine(Met) aminopeptidase inhibitor; Diphosphonate; The biological response conditioning agent; Antiproliferation antibodies; Heparanase inhibitors; The inhibitor of the carcinogenic isotype of Ras; The Telomere terminal transferase inhibitor; Proteasome inhibitor; Used promoting agent in the haematological malignancies treatment; Target, reduction or the active compound of inhibition Flt-3; The Hsp90 inhibitor; And Temozolomide (TEMODAL

).

Terminology used here " aromatase inhibitor " refers to a kind of inhibition estrogen production,, suppresses substrate rotex and the testosterone compound to oestrone and estradiol conversion respectively that is.This term comprises steroide without limitation, especially Atamestane, Exemestane and formestane, and non-steroids particularly, especially amino glutethimide, Rogletimide (roglethimide), Racemic pyridoglutethimide, Win-24540, testolactone, KETOKONAZOL (ketokonazole), vorozole, fadrozole, Anastrozole and letrozole.Exemestane for example can for example be carried out commercially available form with trade mark AROMASIN and carry out administration with its commercial form.Formestane for example can for example carry out commercially available form with trade mark LENTARON and carry out administration with its commercial form.Fadrozole for example can for example carry out commercially available form with trade mark AFEMA and carry out administration with its commercial form.Anastrozole for example can for example carry out commercially available form with trade mark ARIMIDEX and carry out administration with its commercial form.Letrozole for example can for example carry out commercially available form with trade mark FEMARA or FEMAR and carry out administration with its commercial form.Amino glutethimide for example can for example carry out commercially available form with trade mark ORIMETEN and carry out administration with its commercial form.The combination of the present invention that comprises the agent of aromatase inhibitor based chemotherapy is particularly useful for treating the tumour of hormone receptor positive, for example mammary tumor.

Terminology used here " antiestrogen " refers to the compound of antagonism estrogen effect under the estrogen receptor level.This term comprises tamoxifen, fulvestrant, raloxifene and RALOXIFENE HCL without limitation.Tamoxifen for example can for example carry out commercially available form with trade mark NOLVADEX and carry out administration with its commercial form.RALOXIFENE HCL for example can for example be carried out commercially available form with trade mark EVISTA and carry out administration with its commercial form.Fulvestrant can be as US 4,659, disclosed prepare like that or it for example can for example carry out commercially available form with trade mark FASLODEX and carry out administration with its commercial form in 516.Comprise the tumour that is particularly useful for treating estrogen receptor positive as the combination of the present invention of the chemotherapeutics of antiestrogen, for example mammary tumor.

Terminology used here " antiandrogen " refers to any material that can suppress androgenic biological action and comprises bicalutamide (CASODEX) without limitation, and it for example can be as US4, and disclosed such class is prepared in 636,505.

Terminology used here " gonadorelin agonist " comprises abarelix, goserelin and acetate goserelin without limitation.Goserelin is at US 4,100, carried out open in 274 and for example can for example carry out commercially available form with trade mark ZOLADEX and carry out administration with its commercial form.Abarelix for example can be as US 5,843, disclosedly in 901 prepares like that.

Terminology used here " topoisomerase I inhibitor " comprise without limitation Hycamtin, gefitinib, Rinotecan, camptothecine with and analogue, 9-nitrocamptothecin and macromole camptothecine conjugates PNU-166148 (compd A 1 among the WO99/17804).Rinotecan for example can for example carry out commercially available form with trade mark CAMPTOSAR and carry out administration with its commercial form.Hycamtin for example can for example carry out commercially available form with trade mark HYCAMTIN and carry out administration with its commercial form.

Terminology used here " topoisomerase II inhibitor " comprises anthracycline antibiotics such as Zorubicin (comprising Liposomal formulation, for example CAELYX), daunorubicin, epirubicin, idarubicin and Nemorubicin, anthraquinones mitoxantrone and losoxantrone and podophyllotoxin lignanoids (podophillotoxines) Etoposide and teniposide without limitation.Etoposide for example can for example be carried out commercially available form with trade mark ETOPOPHOS and carry out administration with its commercial form.Teniposide for example can for example carry out commercially available form with trade mark VM 26-BRISTOL and carry out administration with its commercial form.Zorubicin for example can for example carry out commercially available form with trade mark ADRIBLASTIN or ADRIAMYCIN and carry out administration with its commercial form.Epirubicin for example can for example carry out commercially available form with trade mark FARMORUBICIN and carry out administration with its commercial form.Idarubicin for example can for example carry out commercially available form with trade mark ZAVEDOS and carry out administration with its commercial form.Mitoxantrone for example can for example carry out commercially available form with trade mark NOVANTRON and carry out administration with its commercial form.

Term " microtubule activator " refers to microtubule stabilizer, microtubule destabilizer and tubulin polymerization inhibitor, it comprises taxanes without limitation, for example taxol and Docetaxel, catharanthus alkaloid, vinealeucoblastine(VLB) for example, especially Vinblastine sulphate, vincristine(VCR), especially vincristine sulphate and vinorelbine, discodermolide (discodermolides), colchicine and esperamicin with and derivative, for example epothilone B or derivatives thereof.Taxol for example can be with its commercial form, and for example TAXOL carries out commercially available form and carries out administration.Docetaxel for example can for example carry out commercially available form with trade mark TAXOTERE and carry out administration with its commercial form.Vinblastine sulphate for example can for example carry out commercially available form with trade mark VINBLASTIN R.P. and carry out administration with its commercial form.Vincristine sulphate for example can for example carry out commercially available form with trade mark FARMISTIN and carry out administration with its commercial form.Discodermolide for example can be as US 5,010, disclosedly in 099 obtains like that.Also be included in disclosed esperamicin derivatives among WO 98/10121, US 6,194,181, WO 98/25929, WO98/08849, WO 99/43653, WO 98/22461 and the WO 00/31247.Especially preferred is Epothilones A and/or B.

Terminology used here " alkylating agent " comprises endoxan, ifosfamide, melphalan or nitrosourea (BCNU or Gliadel) without limitation.Endoxan for example can for example carry out commercially available form with trade mark CYCLOSTIN and carry out administration with its commercial form.Ifosfamide for example can for example carry out commercially available form with trade mark HOLOXAN and carry out administration with its commercial form.

Term " histone deacetylase inhibitors " or " hdac inhibitor " refer to the inhibition of histone deacetylase and have the compound of antiproliferative activity.It is included in disclosed compound among the WO 02/22577, especially N-hydroxyl-3-[4-[[(2-hydroxyethyl) [2-(1H-indol-3-yl) ethyl]-amino] methyl] phenyl]-2E-2-acrylamide, N-hydroxyl-3-[4-[[[2-(2-Methyl-1H-indole-3-yl)-ethyl]-amino] methyl] phenyl]-the 2E-2-acrylamide with and pharmaceutically useful salt.It especially also comprises octanedioyl aniline hydroxamic acid (SAHA).

Term " antineoplastic antimetabolite " comprises 5 FU 5 fluorouracil (5-FU) without limitation; Capecitabine; Gemcitabine; The agent of DNA demethylation is as 5-azepine cytidine and Decitabine; Methotrexate; Edatrexate; With antifol such as pemetrexed.Capecitabine for example can for example carry out commercially available form with trade mark XELODA and carry out administration with its commercial form.Gemcitabine for example can for example carry out commercially available form with trade mark GEMZAR and carry out administration with its commercial form.Also comprise the monoclonal antibody trastuzumab, it for example can for example carry out commercially available form with trade mark HERCEPTIN and carry out administration with its commercial form.

Terminology used here " platinic compound " comprises carboplatin, cis-platinum (cis-platin), cis-platinum (cisplatinum) and oxaliplatin without limitation.Carboplatin for example can for example carry out commercially available form with trade mark CARBOPLAT and carry out administration with its commercial form.Oxaliplatin for example can for example carry out commercially available form with trade mark ELOXATIN and carry out administration with its commercial form.

Terminology used here " target/reduction albumen or the active compound of lipid kinase and other anti-angiogenic compounds " comprises without limitation: protein tyrosine kinase and/or Serine and/or threonine kinase enzyme inhibitors or lipid kinase inhibitors, for example:

A) target, reduction or the active compound of inhibition Thr6 PDGF BB-acceptor (PDGFR), as target, reduction or the active compound of inhibition PDGFR, especially the compound that suppresses pdgf receptor, for example N-phenyl-2-pyrimidine-amine derivatives, for example imatinib (imatinib), SU101, SU6668 and GFB-111;

B) target, reduce or be suppressed to the active compound of fibroblast growth factor-acceptor (FGFR);

C) target, reduction or suppress the active compound of IGF-1 1 (IGF-1R) especially suppress the compound of IGF-1R receptor active, as disclosed compound in WO 02/092599;

D) target, the active compound of reduction or inhibition Trk receptor tyrosine kinase family;

E) target, the active compound of reduction or inhibition Axl receptor tyrosine kinase family;

F) compound of target, reduction or inhibition c-Met receptor active;

G) target, reduction or the active compound of inhibition c-Kit receptor tyrosine kinase (part of PDGFR family), as target, the active compound of reduction or inhibition c-Kit receptor tyrosine kinase family, especially the compound that suppresses the c-Kit acceptor, for example imatinib;

H) target, reduction or suppress the c-Abl member of family with and the active compound of gene fusion product (for example BCR-Abl kinases), as target, reduction or suppress the c-Abl member of family with and the compound of gene fusion its lytic activity, for example N-phenyl-2-pyrimidine-amine derivatives, for example imatinib; PD180970; AG957; NSC 680410; Or derive from the PD173955 of ParkeDavis;

I) Raf family, MEK, SRC, JAK, FAK, PDK and the member of Ras/MAPK family of target, reduction or arrestin kinase c (PKC) member and serine/threonine kinase or PI (3) kinases family or the kinases family relevant and/or cyclin-member's of dependant kinase family (CDK) active compound and especially at US 5 with PI (3)-kinases, 093, disclosed star spore alkali derivant, for example midostaurin in 330; Other examples for compounds comprises for example UCN-01, Safingol, BAY 43-9006, bryostatin 1, Perifosine; Yi Mofuxin; RO 318220 and RO 320432; GO 6976; Isis 3521; LY333531/LY379196; Isoquinoline compound is as disclosed those compounds in WO 00/09495; FTIs; PD184352 or QAN697 (a kind of P13K inhibitor);

J) compound of target, reduction or arrestin-tyrosine kinase activity is as imatinib mesylate (GLIVEC/GLEEVEC) or tyrphostin (tyrphostin).Preferably a kind of lower molecular weight of tyrphostin (Mr＜1500) compound or its pharmaceutically useful salt, especially be selected from benzylidene propane dinitrile class or S-aryl phenylpropyl alcohol dintrile or Double bottom thing (bisubstrate) quinolines, more particularly any Tyrphostin A23/RG-50810 that is selected from; AG 99; TyrphostinAG 213; Tyrphostin AG 1748; Tyrphostin AG 490; Tyrphostin B44; Tyrphostin B44 (+) enantiomer; Tyrphostin AG 555; AG 494; TyrphostinAG 556, AG957 and Adaphostin (4-{[(2,5-dihydroxy phenyl) methyl] amino }-phenylformic acid diamantane ester; NSC 680410, compound Adaphostin); With

K) target, reduce or suppress receptor tyrosine kinase the epidermal growth factor subfamily (all-or the EGFR of heterodimer form, ErbB2, ErbB3, ErbB4) active compound, as target, reduce or the active compound of inhibition epidermal growth factor receptor family, especially suppress the EGF receptor tyrosine kinase member of family, EGF acceptor for example, ErbB2, ErbB3 and ErbB4 or and EGF or EGF associated ligands bonded compound, albumen or antibody, and general and concrete disclosed compound in WO 97/02266 particularly, albumen or monoclonal antibody, the compound of embodiment 39 for example, or at EP 0564409, WO 99/03854, EP 0520722, EP 0566226, EP 0787722, EP 0837063, US 5,747,498, WO 98/10767, WO 97/30034, WO 97/49688, WO 97/38983 and especially at WO 96/30347 (compound that for example is called as CP 358774), disclosed compound among WO 96/33980 (for example compound ZD 1839) and the WO 95/03283 (for example compound ZM105180), albumen or monoclonal antibody; For example trastuzumab (HerpetinR), Cetuximab, Iressa, erlotinib (Tarceva ^TM), CI-1033, EKB-569, GW-2016, E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3 or E7.6.3 and disclosed 7H-pyrrolo--[2,3-d] pyrimidine derivatives in WO 03/013541.

The compound of other angiogenesis inhibitor comprise its activity have other mechanism for example with the compound of the irrelevant mechanism of the inhibition of albumen or lipid kinase, for example Thalidomide (THALOMID) and TNP-470.

The compound of target, reduction or arrestin or lipid phosphatase activity has for example inhibitor of phosphatase 1, Phosphoric acid esterase 2A, PTEN or CDC25, for example okadaic acid or derivatives thereof.

The compound of inducing cell atomization for example has vitamin A acid, α-γ-or Delta-Tocopherol or α-γ-or δ-tocotrienol.

Terminology used here " cyclooxygenase inhibitors " comprises the 2-arylamino phenylacetic acid and the derivative of for example Cox-2 inhibitor, the replacement of 5-alkyl without limitation, as celecoxib (CELEBREX), rofecoxib (VIOXX), L-791456, valdecoxib or 5-alkyl-2-arylamino phenylacetic acid, for example 5-methyl-2-(2 '-chloro-6 '-fluoroanilino) phenylacetic acid, Luo Mei former times cloth (lumiracoxib).

Term " mTOR inhibitor " refers to compound such as the sirolimus (Rapamune that suppresses Mammals rapamycin (mTOR) target spot and have antiproliferative activity

), everolimus (Certican ^TM), CCI-779 and ABT578.

Tend to for example comprise with the immunomodulator of The compounds of this invention coupling

The active conditioning agent of-mTOR, for example inhibitor comprises formula

Rapamycin and rapamycin derivative, for example comprise

40-O-alkyl-rapamycin derivative, as 40-O-hydroxyalkyl-rapamycin derivative, as 40-O-(2-hydroxyl)-ethyl-rapamycin (everolimus),

32-deoxidation-rapamycin derivative and 32-hydroxyl-rapamycin derivative, as 32-deoxidation rapamycin,

The rapamycin derivative that 16-O-replaces such as 16-penta-2-alkynyloxy group-32-deoxidation rapamycin, 16-penta-2-alkynyloxy group-32 (S or R)-dihydro-rapamycin, 16-penta-2-alkynyloxy group-32 (S or R)-dihydro-40-O-(2-hydroxyethyl)-rapamycin,

On 40 oxygen bases by the rapamycin derivative of acidylate, 40-[3-hydroxyl-2-(hydroxyl-methyl)-2 Methylpropionic acid for example]-rapamycin (being also referred to as CCI779),

The rapamycin derivative that on 40, is replaced by heterocyclic radical, 40-epi-(tetrazyl)-rapamycin (being also referred to as ABT578) for example,

So-called forms of rapamycin analogs (rapalogs), disclosed material in WO9802441 or WO0114387 for example, 40-O-dimethyl oxygen phosphino--rapamycin for example, comprise AP23573 and

The disclosed compound of title with hundred Li Mosi (biolimus) (hundred profits are not taken charge of A9), comprise 40-O-(2-oxyethyl group) ethyl-rapamycin, with with the disclosed compound of the title of TAFA-93, preferred 40-O-(2-hydroxyl)-ethyl-rapamycin, CCI779, ABT578 or AP23573, more preferably 40-O-(2-hydroxyl)-ethyl-rapamycin (everolimus).

Rapamycin and other rapamycin derivative can carry out administration with optimal dose, for example carry out administration with known rapamycin or rapamycin derivative dosage, for example everolimus can be paramount to 15mg with 0.1mg, as 0.1mg to 10mg. for example the dosage of 0.1mg, 0.25mg, 0.5mg, 0.75mg, 1mg, 2.5mg, 5mg or 10mg carry out administration, for example carry out administration with the form of (can disperse) tablet; For example according to the disease of being treated, weekly dose can comprise high to 70mg.Can give other rapamycin derivative with similar dosage range.

Terminology used here " diphosphonate " comprises etidronic acid (etridonic), clodronic acid, tiludronic acid, pamidronic acid, clinic effect of alendronate, Ibandronic acid, risedronic acid and Zoledronic acid without limitation." etidronic acid " for example can for example carry out commercially available form with trade mark DIDRONEL and carry out administration with its commercial form." clodronic acid " for example can for example carry out commercially available form with trade mark BONEFOS and carry out administration with its commercial form." tiludronic acid " for example can for example carry out commercially available form with trade mark SKELID and carry out administration with its commercial form." pamidronic acid " for example can be with its commercial form, for example with trade mark AREDIA ^TMCarry out commercially available form and carry out administration." clinic effect of alendronate " for example can for example carry out commercially available form with trade mark FOSAMAX and carry out administration with its commercial form." Ibandronic acid " for example can for example carry out commercially available form with trade mark BONDRANAT and carry out administration with its commercial form." risedronic acid " for example can for example carry out commercially available form with trade mark ACTONEL and carry out administration with its commercial form." Zoledronic acid " for example can for example carry out commercially available form with trade mark ZOMETA and carry out administration with its commercial form.

Terminology used here " heparanase inhibitors " refers to target, reduction or suppresses the compound of heparin sulfate degraded.This term comprises PI-88 without limitation.

Terminology used here " biological response properties-correcting agent " refers to lymphokine or interferons material, for example interferon-gamma.

Terminology used here " inhibitor of the carcinogenic isotype of Ras ", for example H-Ras, K-Ras or N-Ras refer to target, reduction or suppress compound " farnesyl transferase inhibitor " for example L-744832, DK8G557 or R115777 (Zarnestra) for example of the carcinogenic activity of Ras.

Terminology used here " Telomere terminal transferase inhibitor " refers to target, reduction or suppresses the active compound of Telomere terminal transferase.Target, reduction or the active compound of inhibition Telomere terminal transferase especially suppress the compound of Telomere terminal transferase acceptor, for example Te Motating (telomestatin).

Terminology used here " methionine(Met) aminopeptidase inhibitor " refers to target, reduction or suppresses the active compound of methionine(Met) aminopeptidase.Target, reduction or the active compound of inhibition methionine(Met) aminopeptidase for example are the bengamide or derivatives thereofs.

Terminology used here " proteasome inhibitor " refers to target, reduction or the active compound of arrestin enzyme body.Target, reduction or the active compound of arrestin enzyme body comprise for example PS-341 and MLN 341.

Terminology used here " matrix metallo-proteinase inhibitor " or (" MMP inhibitor ") comprise that without limitation collagen intends peptide (peptidomimetic) and non-plan peptide (nonpeptidomimetic) inhibitor, tetracycline derivant, for example hydroxamate intend peptide (peptidomimetic) inhibitor Batimastat with and analogue Marimastat (BB-2516), prinomastat (AG3340), Mei Tasita (metastat) (NSC 683551) BMS-279251, BAY 12-9566, FAA211, MMI270B or the AAJ996 of good to eat clothes utilization.

Terminology used here " in the treatment of haematological malignancies used promoting agent " comprises FMS-sample tyrosine kinase inhibitor for example target, reduction or suppress the active compound of Flt-3 without limitation; Interferon, rabbit, 1-b-D-arabinofuranosyl adenin base cytosine(Cyt) (ara-c) and bisulfan; And ALK inhibitor for example target, reduction or suppress the compound of Nucleophosmin-anaplastic lymphoma kinase.

Term " target, reduction or the active compound of inhibition Flt-3 " especially suppresses compound, albumen or the antibody of Flt-3, for example PKC412, midostaurin, star spore alkali derivant, SU11248 and MLN518.

Terminology used here " HSP90 inhibitor " comprises target, reduction without limitation or suppresses the compound of the intrinsic atpase activity of HSP90; By the degraded of ubiquitin protein enzyme body approach, target, reduction or the proteic compound of inhibition HSP90 client.Compound, albumen or the antibody that target, reduction or the compound that suppresses the intrinsic atpase activity of HSP90 especially suppress the atpase activity of HSP90 is 17-allyl amino, 17-demethoxylation geldanamycin (17AAG), geldanamycin derivant for example; The compound that other are relevant with geldanamycin; Radicicol and hdac inhibitor.

Terminology used here " antiproliferation antibodies " comprises trastuzumab (Herceptin without limitation ^TM), trastuzumab-DM1, rhuMAb-VEGF (Avastin ^TM), Rituximab (Rituxanr

), PRO64553 (anti-CD 40) and 2C4 antibody.Multi-specificity antibody and antibody fragment that antibody refers to for example complete monoclonal antibody, polyclonal antibody, formed by at least 2 complete antibody are as long as it shows required biologic activity.

For the treatment of acute myeloid leukaemia (AML), the compound of formula I and the leukemia therapy of standard can be united use, especially with the treatment coupling that is used for the treatment of AML.The compound of formula I particularly can and/or be used for the treatment of other medicines such as daunorubicin, Dx, Ara-C, VP-16, teniposide, mitoxantrone, idarubicin, carbon platinum and the PKC412 Combined Preparation of AML with for example farnesyl transferase inhibitor.

The structure of the promoting agent of determining with code, generic name or trade(brand)name can derive from current edition or the database of standard outline " the Merck index (The Merck Index) ", for example Patents International (for example IMS World Publications).

Can as described in the document listed above, be prepared and administration as prior art with the above-claimed cpd of the compound coupling of formula I.

Can also be advantageously with compound and the known methods of treatment coupling of formula I, for example can be with the administration or the especially radiation coupling of compound and the hormone of formula I.

The compound of formula I particularly can be used as radiosensitizer, is particularly useful for treating the tumour to the susceptibility difference of radiotherapy.

" associating " refers to be arranged in a kind of fixed combination of dosage unit form, the medicine box that perhaps is used for Combined Preparation, the compound of its Chinese style I and associating companion can be simultaneously by administration independently or especially making said combined partner capable show in for example synergistic timed interval of cooperation administration respectively or can use the dosage regimen of representing its any combination.

Embodiment

Come the present invention is carried out non-limitative illustration with the following examples.

Temperature is degree centigrade being that unit is measured.Unless stated otherwise, otherwise this reaction at room temperature carry out.

R among the TLC _fRatio between the distance that move in distance that each material of value representation moves and eluent forward position.The R of TLC _fValue is at 5 * 10cm TLC plate (silica gel F ₂₅₄, Merck, Darmstadt, Germany) upward measure; The solvent systems of mark is as follows in an embodiment:

^*10% methyl alcohol/90% methylene dichloride (CH ₂Cl ₂)

^*50% hexane/50% ethyl acetate

^{* *}100% methylene dichloride (CH ₂Cl ₂)

Unless stated otherwise, otherwise to analyze the HPLC condition as follows:

Post: Column Engineering, Inc., Matrix, 3 μ m C18 150 * 4.6mm (Lot#205)

With UV be absorbed in 215 and 254nm under detect.Column temperature is 35 ℃ and retention time (t _R) minute being that unit provides.Flow velocity: 1mL/min.

Gradient: water (0.1%TFA)/acetonitrile (0.1%TFA)=98/2,1 minute, to 100% acetonitrile (0.1%TFA), 10 minutes.Stop 2 minutes (total time of operation: 13 minutes) at 100% time

Abbreviation:

The HPLC high performance liquid chromatography

The Isolute of Isolute=International Solvent Technology

HM-N

The mL milliliter

Min minute

MS-ES electrospray mass spectrum

R _fForward position ratio among the TLC

The RT room temperature

The TFA trifluoroacetic acid

The TLC thin-layer chromatography

t _RRetention time

The UV ultraviolet ray

Parent material:

Synthetic aniline makes up the general method of piece (with N-(3-amino-4-methyl-phenyl)-3-trifluoromethyl-benzoyl Amine illustrates):

Compound N shown in the last figure left side-(3-amino-4-methyl-phenyl)-3-trifluoromethyl-benzamide obtains by at room temperature corresponding nitro-compound (N-(4-methyl-3-nitro-phenyl)-3-trifluoromethyl-benzamide) being carried out hydrogenation with Raney nickel in methyl alcohol.Obtain product with high yield.Intermediate nitro-compound (A)---N-(3-nitro-4-methyl-phenyl)-3-trifluoromethyl-benzamide by with 4-methyl-3-nitro-phenyl amine (B) and 3-trifluoromethyl-Benzoyl chloride (C) in methylene dichloride at room temperature and use triethylamine to react to obtain.Obtained intermediate (A) with good productive rate.In document and patent (for example CAS No.30069-31-9), the different aniline category matter of class Sihe has been described before.For coupling, use corresponding chloride of acid.

According to identical operations, with synthesizing reverse 3-amino-benzamide derivatives---3-amino-4-methyl-N-(3-trifluoromethyl-phenyl)-benzamide and 3-amino-N-(4-methoxyl group-3-trifluoromethyl-phenyl)-4-methyl-benzamide by the commercial parent material that obtains accordingly.

Embodiment 1: pyrazolo [1,5-a] Nicotinicum Acidum [2-methyl-5-(3-trifluoromethyl-benzoyl-amido)- Phenyl]-acid amides

With pyrazolo [1,5-a] pyridine-3-dicarbonyl chloride (100mg, 0.55mmol, Maybridge Lot#291259) with N-(3-amino-4-methyl-phenyl)-(163mg 0.55mmol) is dissolved in the 2ml anhydrous methylene chloride 3-trifluoromethyl-benzamide.(93 μ l 0.66mmol) also at room temperature stir this reaction mixture to add triethylamine.After the formation product is complete, product is extracted with this reaction mixture water extinguishing and with methylene dichloride.Removal of solvent under reduced pressure also is adsorbed onto this crude product on the Isolute.Product is carried out purifying (post: Interchrom Puriflash15/35U C18,70g Cartouche with automatic reversed-phase column chromatography; Water+0.1% trifluoroacetic acid and acetonitrile+0.1% trifluoroacetic acid) and it is dry under high-vacuum pump solvent:, obtain the title compound of white solid form.

HPLC:t _R=10.24 minutes; MS-ES:(M+H) +=439; TLC ^*: R _f=0.63

Embodiment 2: pyrazolo [1,5-a] Nicotinicum Acidum [2-methyl-5-(3-trifluoromethyl-phenyl amino formyl Base)-phenyl]-acid amides

Operation identical operations among use and the embodiment 1 just replaces N-(3-amino-4-methyl-phenyl)-3-trifluoromethyl-benzamide to react with 3-amino-4-methyl-N-(3-trifluoromethyl-phenyl)-benzamide.With automatic reversed-phase column chromatography (post: Interchrom Puriflash 15/35U C18,70gCartouche; Solvent: water+0.1% trifluoroacetic acid and acetonitrile+0.1% trifluoroacetic acid) product is separated and it is dry under high-vacuum pump.Obtain the title compound of white solid form.

HPLC:t _R=10.52 minutes; MS-ES:(M+H) +=439; TLC ^*: R _f=0.60

Embodiment 3:6-dimethylamino alkylsulfonyl-pyrazolo [1,5-a] Nicotinicum Acidum [2-methyl-5-(3-fluoroform Base-phenyl amino formyl radical)-phenyl]-acid amides

(200mg 0.35mmol) at room temperature reacted in the 2ml anhydrous pyridine 18 hours with 3-amino-4-methyl-N-(3-trifluoromethyl-phenyl)-benzamide with 6-dimethylamino alkylsulfonyl-pyrazolo [1,5-a] pyridine-3-dicarbonyl chloride.Removal of solvent under reduced pressure.Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=10.76 minutes; MS-ES+:(M+H) +=546; TLC ^*: R _f=0.25

Being prepared as follows of parent material:

Step 3.1:6-dimethylamino alkylsulfonyl-pyrazolo [1,5-a] pyridine-3-dicarbonyl chloride

(200mg 0.57mmol) is dissolved in the 4ml chloroform and adds oxalyl chloride (97 μ l, 1.14mmol with 6-dimethylamino alkylsulfonyl-pyrazolo [1,5-a] Nicotinicum Acidum; Fluka).Should react and reflux 3 hours, under reduced pressure remove volatile matter then.This crude product is directly used in next step.

Step 3.2:6-dimethylamino alkylsulfonyl-pyrazolo [1,5-a] Nicotinicum Acidum

Operation prepares title compound according to document: Yasumitsu Tamura, Yoshio Sumida, Yasuyoshi Miki and Masazumi Ikeda, J.Chem Soc.Perkin 1,1974,406-409.

Step 3.3:6-dimethylamino alkylsulfonyl-pyrazolo [1,5-a] Nicotinicum Acidum ethyl ester

At room temperature, salt of wormwood and ethyl propiolate are joined in the 1-amino-3-dimethylamino alkylsulfonyl-suspension of pyridine sym-toluenesulfonic acid salt in chloroform that is carrying out stirring.For more details, referring to document: Yasumitsu Tamura, Yoshio Sumida, Yasuyoshi Miki and Masazumi Ikeda, J.Chem Soc.Perkin 1,1974,406-409.

Step 3.4:1-amino-3-dimethylamino alkylsulfonyl-pyridine sym-toluenesulfonic acid salt

Title compound be with pyridine derivate and O-sym-trimethylbenzene alkylsulfonyl oxyamine (MSH) etc. molar mixture come synthetic according to general operation; referring to document Y.Tamura; J.Minamikawa; Y.Miki; S.Matsugashita and M.Ikeda; Tett.Lett. (40), 4133-4135,1972.

Embodiment 4:6-dimethylamino alkylsulfonyl-pyrazolo [1,5-a] Nicotinicum Acidum [5-(4-methoxyl group-3-trifluoro Methyl-phenyl amino formyl radical)-2-methyl-phenyl]-acid amides

Use and embodiment 3 described operation identical operations, just replace 3-amino-4-methyl-N-(3-trifluoromethyl-phenyl)-benzamide with 3-amino-N-(4-methoxyl group-3-trifluoromethyl-phenyl)-4-methyl-benzamide.Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=10.26 minutes; MS-ES+:(M+H) +=576; TLC ^*: R _f=0.19.

Embodiment 5:6-phenyl-pyrazole is [1,5-a] Nicotinicum Acidum [2-methyl-5-(3-trifluoromethyl-phenylamino also The base formyl radical)-phenyl]-acid amides

Use and embodiment 3 steps 3.4 are just used 3-phenylpyridine (Fluka) to 3.1 described identical operations in step 3.4.Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=11.99 minutes; MS-ES+:(M+H) +=515; TLC ^*: R _f=0.45.

Embodiment 6:6-phenyl-pyrazole is [1,5-a] Nicotinicum Acidum [2-methyl-5-(3-trifluoromethyl-benzoyl also Base is amino)-phenyl]-acid amides

Use and embodiment 3 steps 3.4 are to 3.1 described identical operations, just in step 3.4, replace 3-amino-4-methyl-N-(3-trifluoromethyl-phenyl)-benzamide with 3-phenylpyridine (Fluka) and N-(3-amino-4-methyl-phenyl)-3-trifluoromethyl-benzamide.Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=11.48 minutes; MS-ES+:(M+H) +=515; TLC ^*: R _f=0.26.

Embodiment 7:6-(4-methoxyl group-phenyl)-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides

With 6-bromo-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides (50mg, 0.15mmol), 4-anisole ylboronic acid (27.9mg, 0.18mmol), chloro-[2 '-(dimethylamino)-2-xenyl]-(two norcamphyl phosphines)-palladium (4.6mg, 7.6 μ mol; Fluka, CAS#359803-53-5) and salt of wormwood (84mg 0.61mmol) is heated to 130 ℃ and reaches 20 minutes in microwave oven in the anhydrous dioxan of 5ml.Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=11.20 minutes; MS-ES+:(M+H) +=358; TLC ^*: R _f=0.51.

Being prepared as follows of parent material:

Embodiment 7.1:6-bromo-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides

Use and embodiment 3 steps 3.4 just in step 3.4, replace 3-amino-4-methyl-N-(3-trifluoromethyl-phenyl)-benzamide with 3-bromopyridine (Fluka) and neighbour-tolyl amine (Fluka) to 3.1 described identical operations.Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=10.15 minutes; MS-ES+:(M+H) +=331; TLC ^{* *}: R _f=0.32.

Embodiment 8:4-[4-(3-neighbour-tolyl formamyl-pyrazolo [1,5-a] pyridine-6-yl)-phenyl]-piperazine Piperazine-1-formic acid uncle-butyl ester

Use and identical operations described in the embodiment 7, just with 4-[4-(4,4,5,5-tetramethyl--[1,3,2] two oxa-boron heterocycle pentane-2-yls)-phenyl]-piperazine-1-formic acid uncle-butyl ester (Maybridge) replacement 4-anisole ylboronic acid.Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=11.52 minutes; MS-ES+:(M+H) +=512; TLC ^*: R _f=0.46.

Embodiment 9:6-(4-morpholine-4-base-phenyl)-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides

Use and identical operations described in the embodiment 7, just replace 4-anisole ylboronic acid with 4-(morpholine-4-yl) phenyl-boron dihydroxide (Maybridge).Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=9.92 minutes; MS-ES+:(M+H) +=413; TLC ^*: R _f=0.30.

Embodiment 10:6-(3-methoxyl group-phenyl)-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides

Use and identical operations described in the embodiment 7, just replace 4-anisole ylboronic acid with 3-anisole ylboronic acid (Aldrich).Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=10.82 minutes; MS-ES+:(M+H) +=358; TLC ^*: R _f=0.65.

Embodiment 11:6-(4-dimethylamino-phenyl)-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides

Use and identical operations described in the embodiment 7, just replace 4-anisole ylboronic acid with 4-(dimethylamino)-phenyl-boron dihydroxide (Aldrich).Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=8.60 minutes; MS-ES+:(M+H) +=371; TLC ^*: R _f=0.57.

Embodiment 12:6-(3,4-dimethoxy-phenyl)-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides

Use and identical operations described in the embodiment 7, just with 3,4-dimethoxy benzene ylboronic acid (Frontier) replaces 4-anisole ylboronic acid.Product separated with automatic column chromatography and it is dry under high-vacuum pump, obtain the title compound of white solid form.

HPLC:t _R=10.22 minutes; MS-ES+:(M+H) +=388; TLC ^*: R _f=0.47.

Embodiment 13: soft capsule

Following such soft gelatin capsule for preparing 5000 each self-contained 0.05g as one of formula I compound described in any one embodiment of front of activeconstituents:

Form

Activeconstituents 250g

2 liters of Lauroglycol

Preparation method: chippy activeconstituents is suspended in Lauroglykol ^*In the wet-milling millstone, grind to produce the granularity of about 1 to 3 μ m in (propylene glycol laurate, Gattefoss é S.A., Saint Priest, France) and with it.Then, with capsule filling machine this mixture of 0.419g is incorporated in the soft gelatin capsule.

Embodiment 14: the tablet that comprises formula I compound

According to standard operation, preparation have form below comprise the tablet of 100mg as one of formula I compound described in any one embodiment of front of activeconstituents:

Form

Activeconstituents 100mg

Crystallization lactose 240mg

Avicel 80mg

PVPPXL 20mg

Aerosil 2mg

Magnesium Stearate 5mg

447mg

Produce: with activeconstituents and solid support material is admixed together and with tabletting machine (Korsch EKO, compressing tablet diameter 10mm) it is compressed.

Avicel

Be Microcrystalline Cellulose (FMC, Philadelphia, USA).PVPPXL is cross-linked polyvinylpyrrolidone (BASF, a Germany).Aerosil

Be silicon-dioxide (Degussa, Germany).

The inhibition of embodiment 15:EphB4 kinase activity

With the pilot system described in the top general description, the kinase whose ability of compound inhibition EphB4 of embodiment 1 and 2 is tested.Find that it has the IC in the scope given in the top general description ₅₀Value (μ mol/l).

Claims (12)

1. formula (I) compound of free or salt form

Wherein

R1 is hydrogen, halogen, sulfamyl, N-C _1-4Alkylsulfamoyl group, N, N-two-C _1-4Alkylsulfamoyl group, be not substituted or substituted alkyl, be not substituted or substituted aryl, be not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical;

R2 and R3 are hydrogen, halogen, C independently of one another _1-4-alkyl, trifluoromethyl, C _1-4-alkoxyl group or cyano group,

X ₁, X ₂, X ₃And X ₄Be that CH or wherein maximum two can be N;

D is N (R6) (preferably), O or S,

Wherein R6 is hydrogen, acyl group or is not substituted or substituted alkyl,

Y ₁Be O, S, NH, CH ₂, N=CH, CH=N or CH=CH;

Y ₂Be C or, if Ra does not exist, then its (also) can be N;

Ra is

If-Y ₂Be that N and R1 are unsubstituted alkyl, substituted alkyl with 5 or more carbon atoms, are not substituted or substituted aryl, are not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical that it does not exist, perhaps,

If-Y ₂Being that C and R1 are unsubstituted alkyl, substituted alkyl with 5 or more carbon atoms, are not substituted or substituted aryl, are not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical, is hydrogen,

If Y ₂Be that C and R1 are hydrogen, halogen, C _1-4-alkyl, sulfamyl, N-C _1-4Alkylsulfamoyl group, N, N-two-C _1-4Alkylsulfamoyl group or unsubstituted aryl are the parts of formula IA

Wherein dotted line is represented and the rest part of formula I molecule (with shown in the wavy line) bonded key,

A be C (=O)-N (R4) or N (R4)-C (=O),

Wherein R4 is hydrogen or is not substituted or substituted alkyl,

Z ₁Be O, S, NH, CH ₂, CH=N, N=CH or CH=CH,

Z ₂Be nitrogen or CH,

Z ₃Be CH or N,

Each R that occurs ₅Be a kind of substituting group independently of one another, and n is 0,1 or 2.

2. the described formula I compound of claim 1, wherein

R1 is hydrogen, halogen, be not substituted or by one or more, preferred maximum three are independently selected from the following C that substituting group replaced ₁-C ₂₀-alkyl: be not substituted or as following defined substituted heterocyclic radical, be not substituted or as following defined substituted cycloalkyl, be not substituted or as following defined substituted aryl, especially phenyl or naphthyl; C ₂-C ₇-alkenyl, C ₂-C ₇Alkynyl, halogen, hydroxyl, C ₁-C ₇Alkoxyl group, C ₁-C ₇Alkoxy-C ₁-C ₇Alkoxyl group, (C ₁-C ₇-alkoxyl group)-C ₁-C ₇Alkoxy-C ₁-C ₇Alkoxyl group, phenoxy group, naphthyloxy, phenyl-or naphthyl-C ₁-C ₇Alkoxyl group; Amino-C ₁-C ₇Alkoxyl group, C ₁-C ₇-alkanoyloxy, benzoyloxy, naphthoyl oxygen base, nitro, cyano group, carboxyl, C ₁-C ₇Alkoxy carbonyl, phenyl-or naphthyl-C ₁-C ₇Alkoxy carbonyl, C ₁-C ₇Alkyloyl, benzoyl, naphthoyl, formamyl, N-be single-or N, the dibasic formamyl of N-, wherein said substituting group is selected from C ₁-C ₇Alkyl and hydroxyl-C ₁-C ₇Alkyl; Amidino groups, guanidine radicals, urea groups, sulfydryl, C ₁-C ₇Alkylthio, thiophenyl or naphthalene sulfenyl, phenyl-or naphthyl-C ₁-C ₇Alkylthio, C ₁-C ₇Alkyl-thiophenyl, C ₁-C ₇Alkyl-naphthalene sulfenyl, halo-C ₁-C ₇Alkyl thiol, C ₁-C ₇Alkyl sulphinyl, phenyl-or naphthyl-sulfinyl, phenyl-or naphthyl-C ₁-C ₇Alkyl sulphinyl, C ₁-C ₇Alkyl-phenyl sulfinyl, C ₁-C ₇Alkyl-naphthyl sulfinyl, sulfo group, C ₁-C ₇Paraffinic hydrocarbons alkylsulfonyl, phenyl-or naphthyl-alkylsulfonyl, phenyl-or naphthyl-C ₁-C ₇Alkyl sulphonyl, C ₁-C ₇Alkyl phenyl alkylsulfonyl, halo-C ₁-C ₇Alkyl sulphonyl, sulfonamido, phenylsulfonamido, amino, N-be single-or N, N-two-[C ₁-C ₇Alkyl, phenyl and/or phenyl-C ₁-C ₇Alkyl]-amino; Wherein as substituting group or be substituted above-mentioned each phenyl or naphthyl of the substituent part of alkyl, also comprise that the phenyl or naphthyl itself that is arranged in phenoxy group or naphthyloxy is not substituted or by one or more, for example maximum 3, preferred 1 or 2 is independently selected from following substituting group and replaces: halogen, halo-C ₁-C ₇Alkyl, hydroxyl, C ₁-C ₇Alkoxyl group, amino, N-be single-or N, N-two-(C ₁-C ₇Alkyl, phenyl, naphthyl, phenyl-C ₁-C ₇Alkyl and/or naphthyl-C ₁-C ₇Alkyl)-amino, nitro, carboxyl, C ₁-C ₇-alkoxy carbonyl, formamyl, cyano group and/or sulfamyl;

Be not substituted or substituted aryl, it is to have to be no more than 20 carbon atoms, especially be no more than the unsaturated carbon loop systems of 16 carbon atoms, it is single-, two-or three rings, it is not substituted, perhaps, in the situation of substituted aryl, it is by one or more, and preferred maximum three, for example one or two is independently selected from following substituting group and replaces: phenyl, naphthyl, phenyl-or naphthyl-C ₁-C ₇-alkyl, hydroxyl-C ₁-C ₇-alkyl, C ₁-C ₇-alkoxy-C ₁-C ₇-alkyl, (C ₁-C ₇-alkoxyl group)-C ₁-C ₇-alkoxy-C ₁-C ₇-alkyl, C ₁-C ₇-alkyloyl-C ₁-C ₇-alkyl, halo-C ₁-C ₇-alkyl is as trifluoromethyl; Phenoxy group-or naphthyloxy-C ₁-C ₇-alkyl, phenyl-or naphthyl-C ₁-C ₇-alkoxy-C ₁-C ₇-alkyl, C ₁-C ₇-alkoxyl group-ketonic oxygen base-C ₁-C ₇-alkyl, phenyl-or naphthyl-C ₁-C ₇-alkoxy-carbonyl oxy-C ₁-C ₇-alkyl, cyano group-C ₁-C ₇-alkyl, C ₂-C ₇-alkenyl, C ₂-C ₇-alkynyl, C ₁-C ₇-alkyloyl, halogen, hydroxyl, C ₁-C ₇-alkoxyl group, C ₁-C ₇-alkoxy-C ₁-C ₇-alkoxyl group, (C ₁-C ₇-alkoxyl group)-C ₁-C ₇-alkoxy-C ₁-C ₇-alkoxyl group, phenoxy group, naphthyloxy, phenyl-or naphthyl-C ₁-C ₇-alkoxyl group, amino-C ₁-C ₇-alkoxyl group, C ₁-C ₇-alkanoyloxy, benzoyloxy, naphthoyl oxygen base, nitro, amino, list-, two-or the three-amino that replaces, wherein amino substituting group is independently selected from C ₁-C ₇-alkyl, C ₁-C ₇-alkyloyl, C ₁-C ₇-alkanesulfonyl, phenyl, naphthyl, phenyl-C ₁-C ₇-alkyl and naphthyl-C ₁-C ₇-alkyl; Cyano group, carboxyl, C ₁-C ₇-alkoxy carbonyl, phenyl-or naphthyl-C ₁-C ₇-alkoxy carbonyl, benzoyl, naphthoyl, formamyl, N-be single-or N, the dibasic formamyl of N-, wherein said substituting group is selected from C ₁-C ₇-alkyl and hydroxyl-C ₁-C ₇-alkyl; Amidino groups, guanidine radicals, urea groups, sulfydryl, C ₁-C ₇-alkylthio, thiophenyl or naphthalene sulfenyl, phenyl-or naphthyl-C ₁-C ₇-alkylthio, C ₁-C ₇-alkyl-thiophenyl, C ₁-C ₇-alkyl-naphthalene sulfenyl, halo-C ₁-C ₇-alkyl thiol, C ₁-C ₇-alkyl sulphinyl, phenyl-or naphthyl-sulfinyl, phenyl-or naphthyl-C ₁-C ₇-alkyl sulphinyl, C ₁-C ₇-alkyl-phenyl sulfinyl, C ₁-C ₇-alkyl-naphthyl sulfinyl, sulfo group, C ₁-C ₇-alkanesulfonyl, phenyl-or naphthyl-alkylsulfonyl, phenyl-or naphthyl-C ₁-C ₇-alkyl sulphonyl, C ₁-C ₇-alkyl phenyl alkylsulfonyl, halo-C ₁-C ₇-alkyl sulphonyl, sulfonamido, phenylsulfonamido, tetramethyleneimine-1-base, piperidines-1-base, by amino or N-single-or N, N-two-[C ₁-C ₇-alkyl, phenyl and/or phenyl-C ₁-C ₇-alkyl)-the amino piperidines-1-base that replaces, be not substituted or N-C by the ring carbon atom bonded ₁-C ₇Piperidyl, piperazine-1-base, C that-alkyl replaces ₁-C ₇-alkylpiperazine-1-base, morpholine-4-base or thiomorpholine-4-base; Each phenyl or naphthyl of a wherein above-mentioned substituent part as substituting group or substituted aryl itself is not substituted or by one or more, and is for example high to 3, and preferred 1 or 2 is independently selected from following substituting group and replaces: halogen, halo-C ₁-C ₇-alkyl, hydroxyl, C ₁-C ₇-alkoxyl group, amino, N-be single-or N, N-two-(C ₁-C ₇-alkyl, phenyl, naphthyl, phenyl-C ₁-C ₇-alkyl and/or naphthyl-C ₁-C ₇-alkyl) amino, nitro, carboxyl, C ₁-C ₇-alkoxy carbonyl, formamyl, cyano group and/or sulfamyl;

Be not substituted or substituted cycloalkyl, wherein cycloalkyl is to have 3 to 16, the more preferably saturated monocycle of 3 to 9 ring carbon atoms or bicyclic hydrocarbon base and by one or more, preferred one to three is independently selected from about the described substituting group of substituted aryl and replaces or be not substituted;

Or be not substituted or substituted heterocyclic radical, heterocyclic radical wherein is the heterocyclic group of unsaturated, saturated or fractional saturation, and is monocycle or two ring or trinucleated rings; It has 3 to 24, and more preferably 4 to 16,4 to 10 annular atomses most preferably; Wherein one or more, preferred one to four, especially one or two carboatomic ring atom heteroatoms of being selected from nitrogen, oxygen and sulphur replaces, and coupling collar preferably has 4 to 12, especially 5 to 7 annular atomses; This heterocyclic group is not substituted or by one or more, especially 1 to 3 be independently selected from above " substituted aryl " down defined substituting group replace; And wherein heterocyclic radical especially is selected from Oxyranyle, aziridinyl, the ethylenimine base, 1,2-oxygen thia cyclopentyl, thienyl, furyl, tetrahydrofuran base, pyranyl, the thiapyran base, thianthrenyl, isobenzofuran-base, benzofuryl, chromenyl, the 2H-pyrryl, pyrryl, pyrrolinyl, pyrrolidyl, imidazolyl, imidazolidyl, benzimidazolyl-, pyrazolyl, pyrazinyl, pyrazolidyl, thiazolyl, isothiazolyl, dithiazole base oxazolyl isoxazolyl, pyridyl, pyrazinyl, pyrimidyl, piperidyl, piperazinyl, pyridazinyl, morpholinyl, thio-morpholinyl, (S-oxo or S, the S-dioxo)-thio-morpholinyl, the indolizine base, pseudoindoyl, the 3H-indyl, indyl, benzimidazolyl-, cumaryl, indazolyl, triazolyl, tetrazyl, purine radicals, the 4H-quinolizinyl, isoquinolyl, quinolyl, tetrahydric quinoline group, tetrahydro isoquinolyl, decahydroquinolyl, the octahydro isoquinolyl, benzofuryl, dibenzofuran group, benzothienyl, the dibenzothiophene base, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolyl, quinazolyl, the cinnolines base, pteridyl, carbazyl, the β-Ka Lin base, phenanthridinyl, acridyl, perimidinyl, the phenanthroline base, the furazan base, phenazinyl, phenothiazinyl phenoxazinyl, chromenyl, different chromanyl and chromanyl, these groups are not substituted separately or are selected from the substituent substituting group of mentioning about the aryl that replaces by one to three and replace, preferably be selected from low alkyl group by one or two, especially methyl or tert-butyl, lower alkoxy, especially the group of methoxyl group and halogen, especially bromine or chlorine replaces;

R2 and R3 are hydrogen, halogen, C independently of one another ₁-C ₄-alkyl, trifluoromethyl, C ₁-C ₄-alkoxyl group or cyano group,

X ₁, X ₂, X ₃And X ₄CH preferably, perhaps wherein maximum two can be N;

D is N (R6) (preferably), O or S,

Wherein R6 is hydrogen, C ₁-C ₇-alkyloyl, benzoyl, naphthoyl, phenyl-C ₁-C ₇-alkyloyl, naphthyl-C ₁-C ₇-alkyloyl, phenyl sulfonyl or lower alkane alkylsulfonyl or be not substituted or as top defined substituted alkyl;

Y ₁Be O, S, NH, CH ₂, N=CH, CH=N or CH=CH preferably;

Y ₂Preferably C or, if Ra does not exist, then can be N;

Ra is

If-Y ₂Be that N and R1 are C ₅-C ₂₀-alkyl, as top defined substituted alkyl, be not substituted or as top defined substituted aryl, be not substituted or as top defined substituted cycloalkyl be not substituted or as top defined substituted heterocyclic radical, it does not exist, perhaps

If-Y ₂Be C, R1 is C ₅-C ₂₀-alkyl, as top defined substituted alkyl, be not substituted or as top defined substituted aryl, be not substituted or as top defined substituted cycloalkyl be not substituted or as top defined substituted heterocyclic radical, be hydrogen

-or, if Y ₂Be that C and R1 are hydrogen, halogen or C ₁-C ₄-alkyl then is the part of formula IA

Wherein dotted line is represented the rest part bonded key with formula I molecule,

A be C (=O)-N (R4) or N (R4)-C (=O),

Wherein R4 is hydrogen or is not substituted or substituted alkyl,

Z ₁Be O, S, NH, CH ₂, CH=N, N=CH or preferred CH=CH,

Z ₂Be nitrogen or preferred CH,

Z ₃Be CH or N,

Each R that occurs ₅Be to be selected from following substituting group: C independently of one another ₁-C ₇-alkyl, halogen, halo-C ₁-C ₇-alkyl, C ₁-C ₇-alkyloyl, hydroxyl, C ₁-C ₇The amino of-alkoxyl group, nitro, amino, list-or two-replace, amino substituting group wherein is independently selected from C ₁-C ₇-alkyl and C ₁-C ₇-alkyloyl, cyano group, carboxyl, C ₁-C ₇-alkoxy carbonyl, formamyl, N-be single-or N, N-two-(C ₁-C ₇-alkyl)-formamyl, amidino groups, guanidine radicals, urea groups, lower alkylthio, sulfo group, the C that replace ₁-C ₇-alkanesulfonyl and sulfonamido; And n is 0,1 or 2.

3. the described formula I compound of claim 1, wherein

R1 is hydrogen, halogen or C ₁-C ₄-alkyl;

R2 and R3 are hydrogen, C independently of one another ₁-C ₄-alkyl, halogen or cyano group;

X ₁, X ₂, X ₃And X ₄Each is CH naturally;

D is N (R6), and wherein R6 is hydrogen, C ₁-C ₇-alkyloyl, C ₁-C ₇-alkyl or phenyl-C ₁-C ₇-alkyl;

Y ₁Be CH=CH;

Y ₂Be C;

Ra is the part of formula IA

Wherein dotted line is represented the rest part bonded key with formula I molecule,

A be C (=O)-N (R4) or N (R4)-C (=O),

Wherein R4 is hydrogen or C ₁-C ₇-alkyl,

Z ₁Be CH=CH,

Z ₂Be N or CH,

Z ₃Be CH,

Each R that occurs ₅Be to be selected from following substituting group: C independently of one another ₁-C ₇-alkyl, halo-C ₁-C ₇-alkyl, C ₁-C ₇-alkyloyl, hydroxyl, C ₁-C ₇The amino of-alkoxyl group, amino, list-or two-replace, amino substituting group wherein is independently selected from C ₁-C ₇-alkyl and C ₁-C ₇-alkyloyl, cyano group, carboxyl, C ₁-C ₇-alkoxy carbonyl, formamyl and N-list-or N, N-two-(C ₁-C ₇-alkyl)-and the formamyl that replaces, and n is 0,1 or 2.

4. the compound of the described formula I of claim 1, it is selected from

Pyrazolo [1,5-a] Nicotinicum Acidum [2-methyl-5-(3-trifluoromethyl-benzoyl-amido)-phenyl]-acid amides,

Pyrazolo [1,5-a] Nicotinicum Acidum [2-methyl-5-(3-trifluoromethyl-phenyl amino formyl radical)-phenyl]-acid amides,

6-dimethylamino alkylsulfonyl-pyrazolo [1,5-a] Nicotinicum Acidum [2-methyl-5-(3-trifluoromethyl-phenyl amino formyl radical)-phenyl]-acid amides,

6-dimethylamino alkylsulfonyl-pyrazolo [1,5-a] Nicotinicum Acidum [5-(4-methoxyl group-3-trifluoromethyl-phenyl amino formyl radical)-2-methyl-phenyl]-acid amides,

The 6-phenyl-pyrazole is [1,5-a] Nicotinicum Acidum [2-methyl-5-(3-trifluoromethyl-phenyl amino formyl radical)-phenyl]-acid amides also,

The 6-phenyl-pyrazole is [1,5-a] Nicotinicum Acidum [2-methyl-5-(3-trifluoromethyl-benzoyl-amido)-phenyl]-acid amides also,

6-(4-methoxyl group-phenyl)-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides,

4-[4-(3-neighbour-tolyl formamyl-pyrazolo [1,5-a] pyridine-6-yl)-phenyl]-piperazine-1-formic acid uncle-butyl ester,

6-(4-morpholine-4-base-phenyl)-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides,

6-(3-methoxyl group-phenyl)-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides,

6-(4-dimethylamino-phenyl)-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides and

6-(3,4-dimethoxy-phenyl)-pyrazolo [1,5-a] Nicotinicum Acidum neighbour-tolyl acid amides,

In various situations, it can be free form or salt form.

5. comprise the formula I compound of defined free form of claim 1 or pharmaceutical acceptable salt and the pharmaceutical preparation of pharmaceutically acceptable carrier.

6. the diagnosis or the defined free form of claim 1 of therapeutic treatment or the formula I compound of pharmaceutical acceptable salt that are used for animal or human's body.

7. be used for the treatment of protein kinase is regulated the defined free form of claim 1 of the disease that response is arranged or the formula I compound of pharmaceutical acceptable salt.

8. the formula I compound of defined free form of claim 1 or pharmaceutical acceptable salt is regulated disease that response is arranged or preparation in treatment and is used for the treatment of purposes in the pharmaceutical preparation of protein kinase being regulated the disease that response is arranged to protein kinase.

9. the described purposes of claim 8, wherein saidly protein kinase is regulated the disease of response is arranged is that one or more are selected from the disease that the inhibition of one or more protein tyrosine kinases is had response, described protein tyrosine kinase is selected from c-src kinases, VEGF-receptor kinase (for example KDR and Flt-1), RET-receptor kinase and/or Ephrin receptor kinase, for example EphB2 kinases, EphB4 kinases or associated kinase.

10. the described purposes of claim 8 or claim 9, wherein the disease of being treated is that one or more are selected from following disease:

Proliferative disease, leukemia for example, especially chronic granulocytic leukemia (CML) or ALL, hyperplasia, fibrosis, as liver cirrhosis, vasculogenesis, psoriatic, atherosclerosis, especially artery or transplant after atherosclerosis, proliferation of smooth muscle in the blood vessel, restenosis as narrow or postangioplasty, tumour or Cancerous disease, especially optimum or especially malignant tumour or Cancerous disease, it more preferably is solid tumor, the cancer of the brain for example, kidney, liver cancer, adrenal carcinoma, bladder cancer, breast cancer, cancer of the stomach, ovarian cancer, colorectal carcinoma, the rectum cancer, prostate cancer, carcinoma of the pancreas, lung cancer, cervical cancer, carcinoma of vagina, carcinoma of endometrium, thyroid carcinoma, sarcoma, glioblastoma, multiple myeloma or gastrointestinal cancer, colorectal adenomas, melanoma, or the tumour of head and neck, the for example head and the squamous cell carcinoma of neck, mesangial cell-proliferative disease, malignant pleural mesothelioma, lymphoma, multiple myeloma, tumorigenesis, especially the tumorigenesis of epithelium characteristic is for example in the situation of mastocarcinoma; Hyperproliferative epidermal (not being cancer), especially psoriatic; Hyperplasia of prostate; Kaposi's sarcoma, thrombosis, scleroderma; Disease of immune system; Wherein relate at least a albumen (preferred tyrosine) kinases, especially be selected from maincenter that the kinase whose signal of the protein tyrosine kinase preferably mentioned transmits and diseases in peripheral nerve system, retinopathy, as diabetic retinopathy, neovascular glaucoma or macular degeneration, obesity, hemangioblastoma, vascular tumor, diabetic nephropathy; Malignant nephrosclerosis; Inflammatory diseases is as rheumatoid or rheumatic inflammatory disease, especially sacroiliitis, as rheumatoid arthritis, other chronic inflammatory illness, as chronic asthma, endometriosis, Crohn's disease; Hodgkin; Glomerulonephritis; Inflammatory bowel; Embolic microangiopathy syndrome; Transplant rejection, glomerulopathy; Neural tissue injury; Restenosis, for example, the restenosis of stent-induced; Wherein wish to stimulate or promote neurotization (neuron regeneration; Neurotization) the adjusting to protein kinase such as Eph receptor kinase has situation, disease or the illness of response, as axonal regeneration, or wherein wishes to suppress or reverse neurodegeneration (neuronal degeneration; Neurodegeneration) the adjusting to protein kinase such as Eph receptor kinase has situation, disease or the illness of response, for example Spinal injury, hypoxemia situation, traumatic brain injury, infarct, apoplexy, multiple sclerosis or other neurodegeneration situation, disease or illness.

11. adjusting has the disease of response, especially protein kinase suppressed that the disease of response, the especially method of disease described in one or more claims 10 are arranged to protein kinase in a treatment, it comprises that the animal or human to such treatment of needs uses the defined free form of claim 1 of significant quantity or the formula I compound of pharmaceutical acceptable salt.

12. a method for preparing the formula I compound of defined free form of claim 1 or salt form, it comprises

A) incite somebody to action wherein R1, X ₁, X ₂, X ₃And X ₄Carboxylic acid or its reactive derivatives suc as formula the defined formula II of I compound

With wherein R2, R3, D, Y ₁, Y ₂With the compound condensation of Ra suc as formula the defined formula III of I compound

Perhaps

B) for R1 wherein be not substituted or substituted alkyl, be not substituted or substituted aryl, be not substituted or substituted cycloalkyl be not substituted or the formula I compound of substituted heterocyclic radical synthetic for, wherein R1 is not substituted or substituted alkyl, is not substituted or substituted aryl (preferably), is not substituted or substituted cycloalkyl or be not substituted or substituted heterocyclic radical (separately by the combination of C-atom) and A are hydroxyl or lower alkoxy or B (A) ₂Be assorted two ring [3.3.1] nonanes of 9-boron or-B (CHCH ₃CH (CH ₃) ₂) ₂The boric acid of formula (IV)

R1-B(A) ₂ (IV)，

With wherein R1, R2, R3, X ₁, X ₂, X ₃, X ₄, Y ₁, Y ₂, D and Ra define suc as formula the compound of I and Hal is a halogen, especially chlorine, iodine or preferably bromine or (perfluorination C ₁-C ₄-alkyl)-compound of the formula V of sulfonyloxy reacts

And if necessary, the compound of formula I is changed into another kind of different formula I compound, the salt of the formula I compound that obtains is changed into free cpds or another kind of different salt, the free formula I compound that obtains is changed into its salt and/or the isomer mixture of the formula I compound that obtains is separated into one isomer.