CN1909908A - Pyrazolo[1,5-a]pyrimidin-7-yl-amine derivatives for the treatment of protein kinase-dependent diseases - Google Patents

Abstract

The present invention relates to the use of pyrazolo[1,5a]pyrimidin-7-ylamine compounds and salts thereof in treating kinase-dependent diseases and the use thereof in preparing pharmaceutical products for treating the diseases, novel pyrazolo[1,5a]pyrimidin-7-ylamine compounds, and methods for preparing novel pyrazolo[1,5a]pyrimidin-7-ylamine compounds.

Description

Pyrazolo [1, the 5-a] pyrimidin-7-yl-amine derivative that is used for the treatment of protein kinase dependent diseases

Summary of the invention

The present invention relates to pyrazolo [1,5-a] pyrimidin-7-yl amine derivatives in treatment protein kinase dependent diseases (protein kinase dependent diseases) purposes or be used to prepare the purposes of the pharmaceutical composition for the treatment of described disease, relate to pyrazolo [1,5-a] pyrimidin-7-yl amine derivatives treats the method for described disease, relate to be used for the treatment of described disease comprise pyrazolo [1,5-a] pharmaceutical preparation of pyrimidin-7-yl amine derivatives, relate to new pyrazolo [1,5-a] pyrimidin-7-yl amine derivatives, relate to and make new pyrazolo [1,5-a] method of pyrimidin-7-yl amine derivatives and pharmaceutical preparation, relate to above-mentioned pyrazolo [1,5-a] purposes or the using method of pyrimidin-7-yl amine derivatives, and/or relate to these pyrazolos that are used for the treatment of animal or human's body [1,5-a] pyrimidin-7-yl amine derivatives.

Background of invention

Existing bibliographical information pyrazolo [1,5-a] pyrimidin-7-yl-amine derivative is the part (people such as S.Selleri for example of benzodiazepine receptor, Bioorg.Med.Chem 7 (12), 2705-11 (1999)), the antagonist of corticotropin-releasing factor (EP 1097709), angiotensin ii receptor antagonist (people such as S.Takeshi for example, Japn.Pharm.Bull.47 (7), 928-38 (1999)), single oxide synthetase inhibitors (JP 10101671), analgesic (WO 9535298), antifungal (EP071792) or antiinflammatory (WO 9218504).

We find that now pyrazolo [1,5-a] pyrimidin-7-yl imines residue also can be used as the template that designs effective inhibitors of kinases.

Consider the disease that a large amount of kinases inhibitors and numerous hypertrophy are relevant with other protein kinases, always existing provides the needs that also therefore are used for the treatment of the newtype chemical compound of relevant disease as kinases inhibitor.

From the possible therapy viewpoint of proliferative disease, required is that superfluous type of compounds is arranged, and its each type is fit to specific protein kinase or protein kinase type, thereby allows to realize specific treatment.Therefore, there are the strong needs of finding to allow this type of specific inhibiting newtype chemical compound.

Summary of the invention

Be surprised to find pyrazoles [1 described here, 5a] pyrimidin-7-yl amines type, especially fall into such noval chemical compound and have the pharmacy favorable properties, allow to suppress the kinases of particular type or kind or group, especially allow to suppress c-Abl, Bcr-Abl, c-Kit, c-Raf, Flt-1, Flt-3, KDR, Her-1, the PDGFR kinases, c-Src, the RET receptor kinase, FGF-R1, FGF-R2, FGF-R3, FGF-R4, liver is joined protein receptor kinases (EphB2 kinases for example, EphB4 kinases and relevant Eph kinases), casein kinase (CK-1, CK-2, G-CK), Pak, ALK, ZAP70, Jak1, Jak2, Axl, Cdk1, cdk4, cdk5, Met, FAK, Pyk2, Syk, insulin receptor kinase, Tie-2 or kinases such as Bcr-Abl, c-Kit, c-Raf, Flt-3, FGF-R3, pdgf receptor, the constitutive activation sudden change (activated kinases) of RET and Met.Pyrazolo described herein [1,5a] pyrimidin-7-yl amines type also suppresses described kinase whose mutant.Except this definite activity, pyrazolo [1,5a] pyrimidin-7-yl amine derivatives also has the advantage that main chain allows multiple substitute mode, wherein said multiple substitute mode provides the extensive probability of accurate adjusting of the specific interaction of realization and target kinase binding site, thereby has opened new promising and specific in various degree inhibitors of kinases is provided.

Consider these activity, chemical compound can be used for this type of kinases, the especially treatment of those kinase whose unusual or overactivity relevant diseases of mentioning.

Detailed Description Of The Invention

In one embodiment, the present invention relates to the purposes that formula (I) compound or pharmaceutically acceptable salt thereof is used for the treatment of protein kinase dependent diseases:

Wherein:

R ₂Be H; That replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; That replace or unsubstituted aliphatic residue; Functional group; Perhaps the replacement by linking group or atom and the connection of pyrazolo [1,5a] pyrimidine-ring or unsubstituted aryl, replacement or unsubstituted heteroaryl or replacement or unsubstituted aliphatic residue; R ₃Can be H, replacement or unsubstituted aryl, replacement or unsubstituted heteroaryl, replacement or unsubstituted aliphatic residue, functional group or can pass through linking group or atom and pyrazolo [1,5a] replacement or the unsubstituted aliphatic residue that connects of pyrimidine-ring, R ₂Or R ₃In be one of at least that replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; Perhaps pass through replacement or unsubstituted heteroaryl or replacement or the unsubstituted aromatic yl residue that a linking group or atom and pyrazolo [1,5a] pyrimidine-ring connect;

A is H, halogen (as bromine), aliphatic series part, functional group, replacement or unsubstituted aryl or replacement or unsubstituted heteroaryl, and R ₁Be H, halogen or low alkyl group.

Embodiment preferred is the purposes that such above-claimed cpd or its officinal salt are used for the treatment of protein kinase dependent diseases:

Wherein:

R ₂Be H; Low alkyl group; Cycloalkyl; Benzyl; Benzothienyl, indyl that low alkyl group replaces; Pyridine radicals or the thiazolyl that replaces by low alkyl group randomly; Low alkyl group sulfoamido and benzene sulfonamido that unsubstituted phenyl or the phenyl that is replaced by one or two substituent group, wherein said substituent group are selected from halogen, hydroxyl, alkoxyl, benzyloxy, cycloalkyl, amino, acetylamino, are replaced by one or two halogen;

R ₃Be H; The low alkyl group that replaces by halogen randomly; Phenyl; Pyridine radicals or  azoles base;

A is the indyl of (a) H, halogen, benzothienyl, pyridine radicals, methyl piperazine phenoxyl, low alkyl group replacement, (b) phenyl that replaces of unsubstituted or one or more substituent groups, wherein said substituent group are selected from list, two or tri-lower alkoxy; Two low alkyl group amidos; Randomly by the dibasic morpholinyl of alkyl; By the piperazinyl that one or more substituent groups replace, wherein said substituent group is selected from: low alkyl group, lower alkoxy, low alkyl group piperazinyl, pyrrolidinyl, dialkyl amido and low-level chain triacontanol; And R ₁Be H.

Protein kinase dependent diseases is preferably a kind of c-Abl of depending on, Bcr-Abl, c-Kit, c-Raf, Flt-1, Flt-3, Her-1, KDR, the PDGFR kinases, c-Src, the RET receptor kinase, FGF-R1, FGF-R2, FGF-R3, FGF-R4, liver is joined protein receptor kinases (EphB2 kinases for example, EphB4 kinases and relevant Eph kinases), casein kinase (CK-1, CK-2, G-CK), Pak, ALK, ZAP70, Jak1, Jak2, Axl, Cdk1, cdk4, cdk5, Met, FAK, Pyk2, Syk, insulin receptor kinase, Tie-2 or kinases such as Bcr-Abl, c-Kit, c-Raf, Flt-3, FGF-R3, pdgf receptor, the disease of the constitutive activation of RET and Met sudden change (activated kinases) and (especially unusual highly express or activated) kinases dependence disease or depend on the activatory disease of kinase pathways perhaps depend on any two or more disease in the kinases of just having mentioned.

Protein kinase dependent diseases more preferably is a kind of disease that depends on c-abl, Flt-3, KDR, c-Src, RET, EphB4, c-kit, cdk1, FGFR-1, c-raf, Her-1, Ins-R or Tek,

Most preferably, disease to be treated is a proliferative disease, be preferably optimum or malignant tumor especially, it more preferably is brain, kidney, liver, the adrenal gland, bladder, breast, stomach (especially gastric tumor), ovary, colon, rectum, prostate, pancreas, lung, vagina, thyroid cancer, sarcoma, glioblastoma, multiple myeloma or human primary gastrointestinal cancers, especially be colon cancer or colorectal adenomas, perhaps cervical region and head tumor, hyperproliferative epidermal is psoriasis especially, prostatic hyperplasia, neoplasia, especially the neoplasia of epithelial character is preferably mastocarcinoma or leukemia.

In another embodiment, the disease that disease to be treated causes for the blood vessel that continues is as psoriasis; Kaposi's sarcoma; Restenosis, for example restenosis of stent-induced; Endometriosis; Crohn disease; Hodgkin; Leukemia; Arthritis is as rheumatoid arthritis; Hemangioma; Fibrohemangioma; Ophthalmic is as diabetic retinopathy and neovascular glaucoma; Kidney disease is as glomerulonephritis; Diabetic nephropathy; Malignant nephrosclerosis; The thrombotic microangiopathy syndrome; Transplant rejection and glomerulopathy; Fibrotic disease is as the sclerosis of liver; The mesangial cell proliferation disease; Arteriosclerosis; Neural tissue injury.

Chemical compound of the present invention also can be used for suppressing the vascular reocclusion after the balloon catheter treatment, after being used for the blood vessel reparation or being used to insert the machinery such as support that keeps vessel open, as immunosuppressant,, be used for the treatment of senile plaque and contact dermatitis as assisting of no scar wound healing.

In another embodiment, the present invention relates to the compound or pharmaceutically acceptable salt thereof of such formula (I):

Wherein:

R ₂Be H; That replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; Aliphatic residue; Functional group; Perhaps the replacement by linking group or atom and the connection of pyrazolo [1,5a] pyrimidine-ring or unsubstituted aryl, replacement or unsubstituted heteroaryl or aliphatic residue; R ₃Can be H, replacement or unsubstituted aryl, heteroaryl, aliphatic residue, functional group or can be by linking group or atom aliphatic residue with the connection of pyrazolo [1,5a] pyrimidine-ring,

R ₂Or R ₃In be one of at least that replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; Perhaps pass through replacement or unsubstituted heteroaryl or replacement or the unsubstituted aromatic yl residue that a linking group or atom and pyrazolo [1,5a] pyrimidine-ring connect; And condition is R ₂With A can not all be unsubstituted phenyl;

A be H, halogen (as bromine), aliphatic series part, functional group, replacement or unsubstituted aryl or heteroaryl; And R ₁Be H, halogen or low alkyl group.

Embodiment preferred is such above-claimed cpd, wherein: R ₂Be H; Low alkyl group; Cycloalkyl; Benzyl; Benzothienyl, indyl that low alkyl group replaces; Pyridine radicals or the thiazolyl that replaces by low alkyl group randomly; Low alkyl group sulfoamido and benzene sulfonamido that unsubstituted phenyl or the phenyl that is replaced by one or two substituent group, wherein said substituent group are selected from halogen, hydroxyl, alkoxyl, benzyloxy, cycloalkyl, amino, acetylamino, are replaced by one or two halogen;

R ₃Be H; The low alkyl group that replaces by halogen randomly; Phenyl; Pyridine radicals or  azoles base;

A is the indyl of (a) H, halogen, benzothienyl, pyridine radicals, methyl piperazine phenoxyl, low alkyl group replacement, (b) phenyl that replaces of unsubstituted or one or more substituent groups, wherein said substituent group are selected from list, two or tri-lower alkoxy; Two low alkyl group amidos; Randomly by the dibasic morpholinyl of alkyl; By the piperazinyl that one or more substituent groups replace, wherein said substituent group is selected from: low alkyl group, lower alkoxy, low alkyl group piperazinyl, pyrrolidinyl, dialkyl amido and low-level chain triacontanol; R ₁Be H; And condition is R ₂With A can not all be unsubstituted phenyl.

Most preferably, the compound is selected from : 3 - {7 - Amino - 3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazol And [1,5-a] pyrimidin-6 - yl } - phenol , 6 - (3 - benzyloxy - phenyl) -3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - phenyl yl ] - Pyrazolo [1,5-a] pyrimidin-7 - yl } - phenol , 6 - (3 - methoxy - phenyl) -3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - benzene Yl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - ( 3,5 - dimethoxy - phenyl) -3 - [ 4 - (4 - methyl - piperazin - 1 - Yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - benzyloxy - phenyl) -3 - [ 4 - (4 - methyl - piperazine -1 - Yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (4 - chloro - phenyl) -3 - [ 4 - (4 - methyl - piperazine triazine-1 - Yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl) -3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - Phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 3 - [4 - (4 - methyl - piperazin-1 - yl ) - phenyl] -6 - phenyl - pyridine Pyrazolo [1,5-a] pyrimidin-7 - amine, 5 - methyl-3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - phenyl] -6 - phenyl - pyridine azole And [1,5-a] pyrimidin-7 - amine, 6 - methyl-3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - phenyl ] -5 - phenyl - pyrazole and [1,5-a] pyrimidin-7 - amine , N-{4 - [7 - amino-3 - (4 - dimethylamino - phenyl ) - pyrazolo [1,5-a] pyrimidine -6 - Yl ] - phenyl } -2,3 - dichloro - benzenesulfonamide 4 - chloro - acid 4 - [7 - amino-3 - (4 - dimethylamino - Phenyl ) - pyrazolo [1,5-a] pyrimidin-6 - yl ] - phenyl ester , 6 - (4 - methoxy - phenyl)-5 - methyl - 3 - phenyl - Pyrazolo [1,5-a] pyrimidin-7 - amine, 3 - (4 - methoxy - phenyl ) -5 - methyl-6 - phenyl - pyrazolo [1,5-a] Pyrimidin-7 - amine, 6 - (4 - bromo - phenyl) -3 - (4 - methoxy - phenyl ) -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - Amine, 6 - (4 - bromo - phenyl)-5 - methyl - 3 - phenyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - ( 2,6 - dichloro- - Phenyl) -3 - phenyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 3 - (3 - methoxy - phenyl)-6 - phenyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 3 - bromo -5 - phenyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - benzo [b] thiophene -3 - Yl -3 - [4 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 3 - (4 - bromine - benzene Yl ) -5 - phenyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 3 - [4 - (4 - methyl - piperazin-1 - yl ) - phenyl ] -6 - thiophene -3 - Yl - pyrazolo [1,5-a] pyrimidin-7 - amine 3 - benzo [b] thiophen-3 - yl -6 - (3 - methoxy - phenyl ) - Pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - benzo -3 - [4 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - methoxy - phenyl) -3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazole And [1,5-a] pyrimidin-7 - amine, 6 - (1 - methyl-1H- indol-3 - yl ) -3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - benzene Yl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (4 - methoxy - phenyl) -3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - Phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (2 - methoxy - phenyl) -3 - [ 4 - (4 - methyl - piperazin-1 - Yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - methoxy - phenyl) -3 - pyridin-3 - yl - pyrazolo [1,5-a] pyrimidin-7 - amine 3 - {7 - Amino - 3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [ 1 , 5-a] Pyrimidin-6 - yl } - phenol , 6 - (3 - benzyloxy - phenyl) -3 - [2 - methoxy-5 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - Pyrazolo [1,5-a] pyrimidin-7 - amine 3 - {7 - Amino - 3 - [2 - methoxy-5 - (4 - methyl - piperazin-1 - yl ) - phenyl Yl ] - pyrazolo [1,5-a] pyrimidin-6 - yl } - phenol , 6 - (2 - benzyloxy - phenyl) -3 - [ 4 - (4 - methyl - piperazin-1 - Yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine 2 - {7 - Amino - 3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - benzene Yl ] - pyrazolo [1,5-a]} pyrimidin-6 - yl } - phenol , 6 - (4 - benzyloxy - phenyl) -3 - [ 4 - (4 - methyl - piperazin - 1 - Yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine 4 - {7 - Amino - 3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - benzene Yl ] - pyrazolo [1,5-a] pyrimidin-6 - yl } - phenol , 6 - (2 - benzyloxy - phenyl ) -3 - [ 3 - (4 - methyl - piperazin-1 - Yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine 2 - {7 - Amino - 3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - benzene Yl ] - pyrazolo [1,5-a] pyrimidin-6 - yl } - phenol , 6 - (4 - benzyloxy - phenyl ) -3 - [ 3 - (4 - methyl - piperazin-1 - Yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine 4 - {7 - Amino - 3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - benzene Yl ] - pyrazolo [1,5-a] pyrimidin-6 - yl } - phenol , 6 - (2 - benzyloxy - phenyl) -3 - [2 - methoxy-5 - (4 - methyl base - Piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine 2 - {7 - Amino - 3 - [2 - methoxy-5 - (4 - armor Yl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-6 - yl } - phenol , 6 - (4 - benzyloxy - phenyl) -3 - [ 2 - Methoxy-5 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine 4 - {7 - Amino- -3 - [2 - methoxy-5 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-6 - yl } - phenol, 6 - (3 - Benzyloxy - phenyl) -3 - [ 1 - methyl-1H- indol-3 - yl ] - pyrazolo [1,5-a] pyrimidin-7 - amine 3 - [7 - amino -3 - ( 1 - methyl-1H- indol-3 - yl ) - pyrazolo [1,5-a] pyrimidin-6 - yl ] - phenol 3 - [7 - amino-3 - pyridyl -3 - Yl - pyrazolo [1,5-a] pyrimidin-6 - yl ] - phenol, 6 - (3 - benzyloxy - phenyl) -3 - (2 - methoxy - phenyl ) - topiramate Yl [1,5-a] pyrimidin-7 - amine 3 - [7 - amino-3 - (2 - methoxy - phenyl ) - pyrazolo [1,5-a] pyrimidine-6 ​​- Yl ] - phenol , 3 - [3 - (4 - methyl - piperazin-1 - yl ) - phenyl] -6 - thiophen-3 - yl - pyrazolo [1,5-a] pyrimidin-7 - base Amine, 3 - (2 - methoxy-5 - (4 - methyl - piperazin-1 - yl ) - phenyl ) -6 - thiophen-3 - yl - pyrazolo [1,5-a] pyrimidine -7 - Amine, 3 - [4 - (4 - methyl - piperazin-1 - yl ) - phenyl] -6 - pyridin-4 - yl - pyrazolo [1,5-a] pyrimidin-7 - Amine, 6 - (3 - amino - phenyl) -3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - Amine, 6 - (3 - amino - phenyl) -3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - Amine, 6 - (2 - amino - phenyl) -3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - Amine, 3 - [4 - (4 - methyl - piperazin-1 - yl ) - phenyl ] -6 - (4 - methyl - thiazol-2 - yl ) - pyrazolo [1,5-a ] pyrimidine Pyridine -7 - amine, 6 - benzo [b] thiophen-3 - yl -3 - [2 - methoxy-5 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - Pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - benzo [b] thiophen-3 - yl -3 - [4 - methoxy - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 3 - (3 - methoxy - phenyl ) -6 - thiophen-3 - yl - pyrazolo [1,5-a] pyrimidin-7 - yl Amine, 6 - (3 - benzyloxy - phenyl ) -3 - (3 - methoxy - phenyl ) - pyrazolo [1,5-a] pyrimidin-7 - amine 3 - [7 - Amino-3 - (3 - methoxy - phenyl ) - pyrazolo [1,5-a] pyrimidin-6 - yl ] - phenol , (4 - {7 - Amino - 3 - [ 4 - (4 - armor Yl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-6 - yl } - phenyl ) - carbamic acid ethyl ester , 6 - (3 - chloro- - Phenyl ) -5 - methyl-3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl ) -5 - methyl-3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidine -7 - base Amine, 6 - (3 - chloro - phenyl) -3 - [2 - methoxy-5 - (4 - methyl - piperazin-1 - yl ) - phenyl ] -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl) -3 - [2 - methoxy - 4 - (4 - methyl - piperazin-1 - yl ) - benzene Yl ] -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - amine 3 - {7 - Amino - 3 - [2 - methoxy - 4 - (4 - methyl - piperazine Triazine-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-6 - yl } - phenol , 6 - (2 - chloro - phenyl) -3 - [ 4 - (4 - methyl base - piperazine Triazine-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (2 - chloro - phenyl) -3 - [ 3 - (4 - methyl - piperazine -1 - Yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (4 - fluoro - phenyl ) -5 - methyl-3 - [4 - ( 4 - methyl - Piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (4 - fluoro - phenyl ) -5 - methyl -3 - [3 - ( 4 - a Yl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl ) -5 - methyl- -3 - {3 - [ 4 - (1 - methyl - piperidin-4 - yl ) - piperazin-1 - yl ] - phenyl} - pyrazolo [1,5-a] pyrimidin-7 - yl amines, 6 - (3 - chloro-4 - fluoro - phenyl ) -5 - methyl-3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - phenyl ] pyrazolo [ 1,5 - a] pyrimidine -7 - Amine, 6 - (3 - chloro-4 - fluoro - phenyl ) -5 - methyl-3 - [ 4 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyridine azole and [1,5-a] pyrimidin-7 - amine, 6 - (3 - bromo - phenyl ) -5 - methyl-3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - phenyl yl ] - pyridine Pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - bromo - benzyl ) -3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazole And [1,5-a] pyrimidin-7 - amine, 6 - (3 - bromo - phenyl) -3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyridine azole and [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl ) -5 - methyl-3 - (3 - morpholin-4 - yl - phenyl ) - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl) -3 - (4 - methoxy - phenyl ) -5 - methyl - pyrazolo [1,5 - -a] Pyrimidin-7 - amine, 6 - (3 - chloro - phenyl ) -3 - [3 - ((2R, 6S) -2,6 - dimethyl - morpholin-4 - yl ) - phenyl ] - 5 - a Yl - pyrazolo [1,5-a] pyrimidin-7 - amine, 2 - (4 - {3 - [7 - amino-6 - (3 - chloro - phenyl ) -5 - methyl - pyrazol and [1,5-a] pyrimidin-3 - yl ] - phenyl } - piperazin-1 - yl ) - ethanol, 6 - benzyl-3 - [ 3 - (4 - methyl - piperazin-1 - yl ) - Phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl) -3 - ( 3,4 - dimethoxy - phenyl ) -5 - Fluoromethyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl) -3 - ( 3,4 - dimethoxy - phenyl ) -5 - Methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro-4 - fluoro - phenyl) -3 - ( 3,4 - dimethoxy - phenyl Yl ) -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro-4 - fluoro - phenyl) -3 - (4 - methoxy - phenyl Yl ) -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (4 - fluoro - phenyl) -3 - (4 - methoxy - phenyl) - Methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 2 - (4 - {3 - [7 - amino-6 - (4 - fluoro - phenyl ) -5 - methyl - pyridine azole And [1,5-a] pyrimidin-3 - yl ] - phenyl } - piperazin-1 - yl ) - ethanol, 6 - ( 3,4 - difluoro - phenyl ) -5 - methyl- -3 - [3 - (4 - methyl - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - ( 3,4 - difluoro- - benzene Yl ) -3 - ( 3,4 - dimethoxy - phenyl ) -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 2 - (4 - {3 - [ 7 - ammonia -6 - (3 - chloro-4 - fluoro - phenyl ) -5 - methyl - pyrazolo [1,5-a] pyrimidin-3 - yl ] - phenyl } - piperazin-1 - yl ) - b Alcohol, 2 - (4 - {3 - [7 - amino-6 - ( 3,4 - difluoro - phenyl ) -5 - methyl - pyrazolo [1,5-a] pyrimidin-3 - yl ] - benzene Yl } - piperazin-1 - yl ) - ethanol, 6 - (3 - chloro - phenyl ) -5 - methyl-3 - [ 3 - (4 - pyrrolidin-1 - yl - piperidin-1 - yl ) - Phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (4 - fluoro - phenyl ) -5 - methyl-3 - [ 3 - (4 - pyrrolidin-1 - base - Piperidin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl) -3 - [ 3 - (4 - diethylamino ammonia Yl - piperidin-1 - yl ) - phenyl ] -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 3 - [3 - (4 - diethylamino - Piperidin-1 - yl ) - phenyl ] -6 - (4 - fluoro - phenyl ) -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (4 - fluoride - Phenyl ) -5 - methyl-3 - [ 3 - (4 - methyl - 4 - oxy - piperazin-1 - yl ) - phenyl ] - pyrazolo [1,5-a] pyrimidin-7 - Amine, 6 - (4 - fluoro - phenyl ) -5 - methyl-3 - [ 3 - (4 - methyl-1 ,4 - dioxo - piperazin-1 - yl ) - phenyl ] - pyrazole And [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl) -3 - [ 3 - (4 - dimethylamino - piperidin-1 - yl ) - phenyl Yl ] -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - ( 3,4 - difluoro - phenyl ) -3 - [ 3 - (4 - dimethyl- ammonia Yl - piperidin-1 - yl ) - phenyl ] -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - phenyl ) -5 - methyl- -3 - ( 3,4,5 - trimethoxy - phenyl ) - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - ( 3,4 - difluoro - phenyl) - Methyl-3 - ( 3,4,5 - trimethoxy - phenyl ) - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - (3 - chloro - benzene Yl ) -3 - (3 - methoxy - phenyl ) -5 - methyl - pyrazolo [1,5-a] pyrimidin-7 - amine, 6 - [7 - amino-3 - ( 3 , 4 - Dimethoxy - phenyl ) - pyrazolo [1,5-a] pyrimidin-6 - yl ] - pyridin-2 - ol, 6 - benzyl-3 - ( 3,4 - dimethoxyphenyl Yl - phenyl ) - pyrazolo [1,5-a] pyrimidin-7 - amine and 3 - ( 3,4 - dimethoxy - phenyl ) -6 - (3 - fluoro - benzyl) - Pyrazolo [1,5-a] pyrimidin-7 - amine .

Also having another embodiment is the purposes of above-claimed cpd in pharmaceutical compositions.

Also having another embodiment is the pharmaceutical composition that comprises above-claimed cpd.

Pharmaceutical composition preferably comprises above-claimed cpd and acceptable drug carrier.

In another embodiment, provide according to above chemical compound and be used for the treatment of purposes in the pharmaceutical composition of kinases dependence disease in preparation.

Another embodiment is the method for preparing above-claimed cpd, and it comprises:

(a) make nitrile A-CH under the condition of organic solvent existing ₂3-oxo-propionitrile that-C ≡ N and Ethyl formate reaction replace with formation,

(b) in organic solvent the 3-oxo-propionitrile of the replacement of condensation step (a) and a hydrazine hydrate to form the 2H-pyrazole-3-yl amine of formula (III):

(c) nitrile that formylated replaces under the condition that has ethylate and Ethyl formate is with the 3-oxo-propionitrile of preparation formula (II):

(d) at the 2H-pyrazole-3-yl amine of 3-oxo-propionitrile that has shrinking type under the condition of organic solvent (II) and formula (III) to form the chemical compound of formula (I).

The invention particularly relates to pyrazolo [1,5a] pyrimidin-7-yl amines or its officinal salt of formula (I),

Wherein:

R ₂Be H; That replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; That replace or unsubstituted aliphatic residue; Functional group; Perhaps the replacement by linking group or atom and the connection of pyrazolo [1,5a] pyrimidine-ring or unsubstituted aryl, replacement or unsubstituted heteroaryl or replacement or unsubstituted aliphatic residue; R ₃Be aryl H, replacement or unsubstituted, replacement or unsubstituted heteroaryl, replacement or unsubstituted aliphatic residue, functional group or can be by linking group or atom aliphatic residue, R with the connection of pyrazolo [1,5a] pyrimidine-ring ₂Or R ₃In be one of at least that replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; Perhaps pass through replacement or unsubstituted heteroaryl or replacement or the unsubstituted aromatic yl residue that a linking group or atom and pyrazolo [1,5a] pyrimidine-ring connect;

A be H, halogen (as bromine), aliphatic series part, functional group, replacement or unsubstituted aryl or replacement or unsubstituted heteroaryl, and R ₁Be H, halogen or low alkyl group, the chemical compound that described compound or pharmaceutically acceptable salt thereof is used for the treatment of protein kinase (especially tyrosine protein kinase) dependence disease or is used to prepare the pharmaceutical composition for the treatment of described disease, use formula (I) is used for the treatment of the method for described disease or is used for the treatment of the pharmaceutical preparation that comprises formula (I) chemical compound of described disease.

The invention particularly relates to such formula (I) compound or pharmaceutically acceptable salt thereof:

Wherein: R ₂Be H; That replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; That replace or unsubstituted aliphatic residue; Functional group; Perhaps the replacement by linking group or atom and the connection of pyrazolo [1,5a] pyrimidine-ring or unsubstituted aryl, replacement or unsubstituted heteroaryl or replacement or unsubstituted aliphatic residue; R ₃Be aryl H, replacement or unsubstituted, replacement or unsubstituted heteroaryl, replacement or unsubstituted aliphatic residue, functional group or can be by linking group or atom aliphatic residue, R with the connection of pyrazolo [1,5a] pyrimidine-ring ₂Or R ₃In be one of at least that replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; Perhaps pass through replacement or unsubstituted heteroaryl or replacement or the unsubstituted aromatic yl residue that a linking group or atom and pyrazolo [1,5a] pyrimidine-ring connect, and condition is R ₂With A can not all be unsubstituted phenyl;

A be H, halogen (as bromine), aliphatic series part, functional group, replacement or unsubstituted aryl or heteroaryl, and R ₁Be H, halogen or low alkyl group,

The chemical compound that described compound or pharmaceutically acceptable salt thereof is used for the treatment of protein kinase (especially tyrosine protein kinase) dependence disease or is used to prepare the pharmaceutical composition for the treatment of described disease, use formula (I) is used for the treatment of the method for described disease or is used for the treatment of the pharmaceutical preparation that comprises formula (I) chemical compound of described disease, is used for the treatment of formula (I) chemical compound of described disease.

The present invention also relates to treat the method for kinases dependence disease, it comprises that to homoiothermic animal especially the people uses pyrazolo [1,5a] the pyrimidin-7-yl amines of formula (I).The invention still further relates to the pyrazolo [1 that comprises formula (I), 5a] pharmaceutical preparation of pyrimidin-7-yl amines, in particular for treatment kinases dependence disease, the pyrazolo [1 that relates to new formula (I), 5a] the pyrimidin-7-yl amines, relate to manufacturing formula (I) pyrazolo [1,5a] pyrimidin-7-yl amines method and be used to prepare their new raw material and intermediate.The present invention also relates to the purposes of chemical compound in the pharmaceutical preparation of preparation treatment kinases dependence disease of formula (I).

Unless otherwise noted, above and hereinafter used generic term preferably have following meaning in this open background: " aryl " is for having the aryl of 6-14 carbon atom, especially phenyl, naphthyl, indenyl, azulene base or anthryl, and do not replace or by one or more, preferably replace by one or two substituent group, wherein substituent group be selected from in the undefined functional group any one and comprise rudimentary halogen; Alkyl; The alkyl that replaces; junior alkyl halides is trifluoromethyl for example; low-grade alkenyl; low-grade alkynyl; low-grade alkane acidyl; lower alkoxy; hydroxyl; another aryl; the hydroxyl of etherificate or esterification; amino; single or dibasic amino; amino low alkyl group; amino lower alkoxy; acetylamino; amidino groups; halogen; nitro; cyano group; cyano-lower alkyl group; carboxyl; esterifying carboxyl group; especially elementary alkoxy carbonyl; methoxycarbonyl for example; positive propoxy carbonyl or isopropoxy carbonyl; alkanoyl; benzoyl; carbamoyl; N-list or N, the dibasic carbamoyl of N-; carbamate; alkyl carbamate; amidino groups; guanidine radicals; carbamide; urea groups; sulfydryl; sulfo group; rudimentary alkylthio; sulfo group amino; sulfoamido; the benzene sulfinyl amido; sulfonic group; phenyl; benzyl; phenoxy group; benzyloxy; the phenyl sulfenyl; phenyl-low alkyl group sulfenyl; the alkyl phenyl sulfenyl; the low alkyl group sulfinyl; the phenyl sulfinyl; phenyl-low alkyl group sulfinyl; the alkyl phenyl sulfinyl; the lower alkyl sulfonyl; benzenesulfonyl; phenyl-low alkyl group sulfonyl; the alkyl phenyl sulfonyl; halogen low alkyl group sulfydryl; halogen-low alkyl group sulfonyl; as trifyl especially; dihydroxy boron (B (OH) ₂), heterocyclic radical and the ring adjacent carbon atom place bonded low-grade alkylidene dioxy base, as methylene-dioxy, phosphono (P (=O) (OH) ₂), hydroxyl-lower alkoxy phosphoryl or two lower alkoxy phosphoryls, carbamoyl, list or two elementary alkyl amido formoxyl, list or two (hydroxy lower alkyl)-carbamoyl, or-NR ₄R ₅, R wherein ₄And R ₅Can be identical or different and be independently H, low alkyl group (for example methyl, ethyl or propyl group); Perhaps R ₄And R ₅Form the 3-8 unit heterocycle (for example piperazinyl, low alkyl group-piperazinyl, azelidinyl, pyrrolidinyl, piperidino, morpholinyl, imidazolinyl) that contains 1-4 nitrogen, oxygen or sulphur atom with the N atom.

Aryl more preferably is phenyl unsubstituted or that replaced by one or two substituent group that is selected from solubilizing group independently, and wherein said solubilizing group is selected from: halogen (as Cl, Br or F), hydroxyl, low alkyl group are (as C ₁-C ₃Low alkyl group or methyl), aryl (as phenyl or benzyl), amino, amino low alkyl group (as dimethylamino), acetylamino, amino lower alkoxy (as amine ethoxylate), the low alkyl group (as fluoro ethyl) that replaces, alkoxyl is (as methoxyl group or benzyloxy, wherein benzyl rings can be replacement or unsubstituted, as 3, the 4-dichloro-benzyloxy), sulfoamino-group, that replace or unsubstituted sulfoamido is (as the benzene sulfinyl amido, chlorobenzene sulfonamide base or 2,3-dichloro-benzenes sulfoamido), that replace or unsubstituted sulfonic group (as chloro-phenylbenzimidazole sulfonic acid base), the carbamide (as 3-three fluoro-methyl-phenyl carbamide or 4-morpholine-4-base-3-trifluoromethyl-phenyl-carbamide) that replaces, alkyl carbamate or carbamates (as ethyl-N-phenyl-carbamate) or-NR ₄R ₅, R wherein ₄And R ₅Can be H, low alkyl group (for example methyl, ethyl or propyl group) identical or different and independently; Perhaps R ₄And R ₅Form the 3-8 unit heterocycle (for example piperazinyl, low alkyl group-piperazinyl, pyridine radicals, indyl, thienyl, thiazolyl, the positive methyl piperazine base of morpholinyl, benzothienyl, azelidinyl, pyrrolidinyl, piperidino or imidazolinyl) that contains 1-4 nitrogen, oxygen or sulphur atom with the N atom, wherein work as R ₄And R ₅When forming heterocycle with the N atom, this heterocycle can be by 1 in the substituent group described herein, 2 or a plurality of replacement, preferably by piperazinyl; Pyrrolidinyl; Alkyl is as methyl; Or hydroxy alkyl, replace as ethanyl.R ₄And R ₅The heterocyclic example that forms with the N atom comprises morpholinyl, and it can be unsubstituted or replaces with methyl or dimethyl; Unsubstituted or with 1,2 or 3 substituent groups, be preferably the piperazinyl that methyl, oxygen base or ethanol replace; Or unsubstituted or with 1,2 or 3 substituent groups, be preferably the piperadinyl that pyrrolidinyl, amine, alkylamine, methyl amine, dialkylamine, dimethyl amine or diethylamide replace.

Heteroaryl is preferably monocycle, but can be two or three rings, and comprises 3-24,4-16 annular atoms preferably, wherein at least one or a plurality of, preferably 1-4 ring carbon atom hetero atom of being selected from O, N or S is alternative.Preferably, heteroaryl is selected from pyridine radicals, indyl, pyrimidine radicals, pyrazolyl,  azoles base, thienyl, benzothienyl, 2H-pyrrole radicals, pyrrole radicals, imidazole radicals, benzimidazolyl, pyrazolyl, indazolyl, purine radicals, pyrazinyl, pyridazinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, 2,3-phthalazinyl, naphthyridine base, quinoxalinyl, quinazolyl, quinnolinyl, indolizine base, 3H-indyl, isoindolyl, different  azoles base, thiazolyl, isothiazolyl, triazolyl, tetrazole radical, furazan base and benzo [d] pyrazoles.

More preferably, heteroaryl is selected from pyridine radicals, indyl, pyrimidine radicals, pyrazolyl,  azoles base, thienyl or benzothienyl.

Heteroaryl can be unsubstituted or is replaced by one or more substituent groups, and wherein said substituent group is selected from the substituent group of aryl defined above, is most preferably replaced by hydroxyl, halogen, low alkyl group such as methyl or lower alkoxy such as methoxy or ethoxy.

Aliphatic refers to any residue based on non-aromatic carbon as used herein.That the example of aliphatic residue comprises replacement or unsubstituted alkyl, cycloalkyl, alkenyl and alkynyl.Alkyl comprises low alkyl group, is preferably to have the alkyl that is no more than 7 carbon atoms, preferably has 1-5 and comprises 5 carbon atoms, for linear or ramose; Preferably, low alkyl group is an amyl group, as n-pentyl, and butyl, as normal-butyl, sec-butyl, isobutyl group, the tert-butyl group, propyl group such as n-pro-pyl or isopropyl, ethyl or methyl.Preferably, low alkyl group is methyl, propyl group or the tert-butyl group.

Cycloalkyl is preferably cyclopenta, cyclohexyl or suberyl, and can be unsubstituted or by one or more, especially replaced by one or two such substituent group, wherein said substituent group is selected from the substituent group of aryl defined above, most preferably by low alkyl group, as methyl; Lower alkoxy is as methoxy or ethoxy; Or hydroxyl replaces.

Alkenyl and alkynyl preferably have and are no more than 7 carbon atoms, preferably have 1-5 and comprise 5 carbon atoms, and can be linear or ramose.

Alkyl, cycloalkyl, alkenyl and alkynyl can be replacement or unsubstituted, and be no more than 3 substituent groups when having when replacing, it comprise any one in other alkyl, cycloalkyl, alkenyl, alkynyl, the aryl substituent defined above or the functional group that defines below in any one.

Halogen is preferably fluorine, chlorine, bromine or iodine, is most preferably fluorine, chlorine or bromine.

Term " connect atom or group " comprises that alkyl is (as-CH as used herein ₂-), oxygen base-O-, ketone-CO-, sulfenyl-S-, sulfonyl-SO ₂-, sulfoxide-SO-, amine-NH-or-NR-, carboxylic acid, alcohol, ester (COO-), amide (--CONR-,-CONHR '-), sulfoamido (SO ₂NH-,-SO ₂NR '-), sulfone (SO ₂-), sulfoxide (SO-), amino, carbamide (NH-CO-NH-,-NR-CO-NH-,-NH-CO-NR-,-NR-CO-NR-), ether (O-), carbamate (NH-CO-O-,-NR-CO-O-) or inverse amide, sulfonamide and ester (NH-CO-,-NR-CO-,-NH-SO ₂-,-NR-SO ₂-,-OOC-).

Term " functional group " comprises carboxylic acid, hydroxyl, halogen, cyano group (CN), ether (OR), ketone (CO-R), ester (COOR), amide (CONH as used herein ₂,-CONHR ,-CONRR '), thioether (SR), sulfonamide (SO ₂NH ₂,-SO ₂NHR ,-SO ₂NRR '), sulfone (SO ₂-R), sulfoxide (SO-R), amine (NHR, NR ' R), carbamide (NH-CO-NH ₂,-NH-CO-NHR), ether (O-R), halogen, carbamate (NH-CO-OR), aldehyde functional group (CHO), also can be then inverse amide, sulfoamido and ester (NH-CO-R ,-NH-SO ₂-R ,-OOC-R), R and R ' can be identical or different, and can be H or any aliphatic as defined above, aryl or heteroaryl moieties.

When plural number is used for chemical compound, salt, pharmaceutical preparation, disease or the like, also mean one chemical compound, salt or the like.Salt is the officinal salt of formula I chemical compound especially.

This type of salt preferably forms as acid-addition salts with organic or inorganic acid from formula (I) chemical compound with basic nitrogen atom, especially officinal salt.Suitable mineral acid is for example halogen acid example hydrochloric acid, sulphuric acid or phosphoric acid.Appropriate organic is for example carboxylic acid; Phosphonic acids; Sulfonic acid or sulfamic acid; Be for example acetic acid; Propanoic acid; Sad; Capric acid; Dodecanoic acid; Glycol acid; Lactic acid; Fumaric acid; Succinic acid; Adipic acid; 1,5-pentanedicarboxylic acid.; Suberic acid; Azelaic Acid; Malic acid; Tartaric acid; Citric acid; Aminoacid is as glutamic acid or aspartic acid; Maleic acid; Hydroxyl-maleic acid; Citraconic acid; Cyclohexane-carboxylic acid; Adamantanecarboxylic acid; Benzoic acid; Salicylic acid; The 4-aminosallcylic acid; Phthalic acid; Phenylacetic acid; Mandelic acid; Cinnamic acid; Methanesulfonic acid or ethyl sulfonic acid; The 2-hydroxyethanesulfonic acid; Ethane-1, the 2-disulfonic acid; Benzenesulfonic acid; The 2-LOMAR PWA EINECS 246-676-2; 1,5-naphthalene-disulfonic acid; 2-, 3-or 4-toluene sulfonic acide; Methylsulfuric acid; Ethyl sulfuric acid; Lauryl sulphate acid; The N-cyclohexylsulfamic acid; The N-methyl-, N-ethyl-or N-propyl group-sulfamic acid; Or other organic Bronsted acids, as ascorbic acid.

There is the negative electricity group, under carboxyl or sulfo group condition, salt also can form with alkali, and wherein said alkali is slaine or ammonium salt for example, as alkali metal salt or alkali salt, for example sodium salt, potassium salt, magnesium salt or calcium salt, or have the ammonia or the suitable ammonium salt of organic amine, as uncle's monoamine, triethylamine or three (2-hydroxyethyl) amine for example, or heterocyclic bases, for example N-ethyl-piperidines or N, N '-lupetazin.

When basic group and acidic-group were present in the same molecule, the chemical compound of formula (I) also can form inner salt.

Be the isolated or purified purpose, also may use not officinal salt, for example picrate or perchlorate.Be therapeutic use, only utilize officinal salt or free cpds (when using), and therefore these are preferred with the pharmaceutical preparation form.

Substantial connection between the chemical compound of consideration free form chemical compound and its salt form, wherein said salt comprises and can be used as for example at the purification of chemical compound, tautomer or tautomers mixture and its salt and those salt of the intermediate in the evaluation, above and the chemical compound of hereinafter referring to especially formula (I) if chemical compound points out to be interpreted as the corresponding tautomer that also refers to these chemical compounds, especially formula I chemical compound as required and advantageously and not in addition; These chemical compounds, the especially tautomers mixture of the chemical compound of formula I; Or any one salt in these.

When mention " chemical compound ..., its tautomer or its salt " etc. the time, this be meant " chemical compound ..., its tautomer, the perhaps salt of chemical compound or tautomer ".

Any asymmetric carbon atom can (R)-, (S)-or (R S)-configuration exists, preferably exists with (R)-or (S)-configuration.Substituent group with annular atoms of saturated bond, if possible, can be suitable-(=Z-) or anti-(=E-) form exists.Therefore chemical compound can exist by isomer mixture, or preferably exists with pure isomer, preferably exists with the diastereomer of enantiomeric pure or pure enantiomer.

The present invention also relates to the prodrug of formula (I) chemical compound, it changes the chemical compound of an accepted way of doing sth (I) in vivo like this.Therefore any chemical compound of referring to formula (I) is interpreted as that as required and advantageously the corresponding prodrug of the chemical compound that also refers to formula (I) is suitably with easily.

The chemical compound of formula (I) has valuable pharmacological character and is used for the treatment of the kinases dependence disease, for example as the medicine for the treatment of proliferative disease.Term " treatment of tyrosine protein kinase dependence disease " refers to disease especially cited below preventative of described disease or preferably therapeutic (comprise alleviate and/or cure) treatment.

When mentioning term " purposes " subsequently, words if not indicated otherwise, this comprises any one or a plurality of following embodiment of the present invention as required and advantageously respectively: the purposes in treatment (especially tyrosine) protein kinase dependent diseases, be used to prepare the purposes of the pharmaceutical composition for the treatment of described disease, pyrazolo [1,5a] pyrimidin-7-yl amine is used for the treatment of the method for described disease, what be used for the treatment of described disease comprises pyrazolo [1,5a] pyrimidin-7-yl amine pharmaceutical preparation and be used for the treatment of pyrazolo [1, the 5-a] pyrimidin-7-yl-amine derivative of described disease.Especially, the disease that the chemical compound of disease to be treated and therefore preferred formula (I) is used is selected from (especially tyrosine) cited below protein kinase dependent, and (" dependency " also is meant " support ", be not only " fully relying on ") disease, especially corresponding proliferative disease, more particularly depend on c-Abl, Bcr-, Abl, c-Kit, c-Raf, Flt-1, Flt-3, KDR, Her-1, the PDGFR kinases, c-Src, the RET receptor kinase, FGF-R1, FGF-R2, FGF-R3, FGF-R4, liver is joined protein receptor kinases (EphB2 kinases for example, EphB4 kinases and relevant Eph kinases), casein kinase (CK-1, CK-2, G-CK), Pak, ALK, ZAP70, Jak1, Jak2, Axl, Cdk1, cdk4, cdk5, Met, FAK, Pyk2, Syk, insulin receptor kinase, Tie-2 or kinases such as Bcr-Abl, c-Kit, c-Raf, Flt-3, FGF-R3, pdgf receptor, the disease of the constitutive activation of RET and Met sudden change (activated kinases) (hereinafter being called " described kinases "), therefore chemical compound can be used for treating the kinases dependence disease, especially depends on described kinase whose disease and (especially unusual high expressed or constitutive activation) described kinases dependence disease or depends on described kinase pathways or the disease of two or more kinase whose any associatings of mentioning.

Most preferred for the chemical compound of formula (I) is used for the treatment of the purposes of the disease that depends on c-abl, Flt-3, KDR, c-Src, RET, EphB4, c-kit, cdk1, FGFR-1, c-raf, Her-1, Ins-R and Tek, and the chemical compound of formula (I) is as the purposes of the inhibitor of c-abl, Flt-3, KDR, c-Src, RET, EphB4, c-kit, FGFR-1, c-raf, cdk1, Her-1, Ins-R and Tek.

The chemical compound experiment of active anticancer in vivo of proof formula (I) is also arranged.

The chemical compound of formula (I) has valuable pharmaceutical properties and useful in the treatment protein kinase dependent diseases, for example as the medicine for the treatment of proliferative disease.

The inhibition of following measure R ET: pFB-GSTX3 produces recombinant baculovirus with baculovirus donor carrier, it expresses the amino acid region 658-1072 (Swiss prot No.Q9BTB0) of the interior kinase domain of kytoplasm of people RET-Men2A (wtRET) corresponding with the wild type kinase domain of RET and RET-Men2B, the different activated mutants that are to activate ring M918T with wtRET of wherein said RET-Men2B.By the cytoplasmic structure domain encoding sequence of PCR from plasmid pBABEpuroRET-Men2A and pBABEpuro RET-Men2B amplification wtRET and RET-Men2B.By making the dna fragmentation of amplification and pFB-GSTX3 carrier be fit to coupled reaction with SaII and KpnI digestion.Connection by these dna fragmentations obtains baculovirus donor plasmids pFB-GX3-RET-Men2A and pFB-GX3-RET-Men2B respectively.

The generation of virus: the transfer vector transfection that will contain kinase domain is in DH10Bac cell line (GIBCO) and be taped against on the selectivity agar plate.The bacterium colony that does not insert fusion sequence (being carried by antibacterial) in viral genome is for blue.Choose plasmid purification step isolated viral DNA (rod granule) from antibacterial that single white colony also passes through standard.Use Cellfectin reagent at 25cm then ²Use viral DNA transfection Sf9 cell or Sf21 (American type culture collection) cell in the bottle.

Measure Sf9 cell middle and small scale protein expression: from the transfectional cell culture, collect the culture medium of combination of syndromes poison and be used for infection to increase its titre.The culture medium that contains virus that obtains after two-wheeled infects is used for large-scale protein expression.For carrying out large-scale protein expression, with 5 * 10 ⁷Individual cell/plating 100cm ²Circular tissue culture plate also infects with the culture medium (approximately 5MOIs) that 1mL contains virus.It is centrifugal 5 minutes to scrape cell and 500 rev/mins from flat board after 3 days.Will be from 10-20 100cm ²Dull and stereotyped cell precipitation be resuspended in the ice-cold lysis buffer of 50mL (25mM tris-HCI, pH 7.5,2mM EDTA, 1%NP-40,1mM DTT, 1mMPMSF).Stirred cell on ice 15 minutes, then 5,000 rev/mins centrifugal 20 minutes.

The purification of GST label protein matter: will (Pharmacia) also use 10mL 25mM tris-HCI in centrifugal cell lysate adding 2mL glutathione agarose post, pH 7.5,2mM EDTA, 1mM DTT, 200mM NaCl washing 3 times.Use (each 1mL) 25mM tris-HCI by 10 times then, pH 7.5, the 10mM reduced glutathione, and 100mM NaCl, 1mM DTT, 10% glycerol eluting GST label protein matter also is stored in-70 ℃.

The measurement of enzymatic activity: containing 15ng GST-wtRET or GST-RET-Men2B protein with purified GST-wtRET or GST-RET-Men2B protein, 20mM tris-HCI, pH 7.5,1mM MnCl2,10mM MgCl2,1mM DTT, 3 μ g/mL gather (Glu, Tyr) 4: 1,1%DMSO, 2.0 μ M ATP (γ-[ ³³P]-ATP 0.1 μ Ci) 30 μ L final volume in carry out the tyrosine protein kinase algoscopy.Under the condition that has or do not exist inhibitor, pass through to measure [γ ³³P] ATP ³³P mixes poly-(Glu, Tyr) mensuration activity in 4: 1.Be determined under the condition as described below and in 96 orifice plates, carried out 15 minutes and passed through to add 20 μ L 125mM EDTA stopping in room temperature.Subsequently 40 μ L reactant mixtures are transferred in advance with methanol and soaked on 5 minutes the Immobilon-PVDF film (Millipore), use 0.5%H then with water rinse ₃PO ₄Soaked into 5 minutes and placed and take off the total chamber of the vacuum that is connected with vacuum source.On the point behind all sample, connect vacuum equipment and with 200 μ L 0.5%H ₃PO ₄Each hole of rinsing.Remove film and on shaking table, use 1.0%H ₃PO ₄Wash 4 times, use washing with alcohol 1 time.Count film after the drying at room temperature, wherein said film places on the PackardTop Count 96 hole supports and adds the Microscint TM (Packard) in 10 μ L/ holes.Calculate IC in the linear regression analysis that the percentage ratio of 4 concentration (being generally 0.01,0.1 and 10 μ M) suppresses in duplicate by each chemical compound ₅₀Value.The protein kinase activity of a unit is defined as 37 ℃ of 1 nanomole ³³P ATP is from [γ ³³P] ATP transfers to substrate protein white matter/minute/milligram protein.

IC50 calculates

On the Immobilon film, import 3 * 4 μ L and stop measuring, without washing

Background (3 holes) is used H ₂O measures and without enzyme

Positive control (4 holes) 3%DMSO rather than chemical compound

Criticize contrast (1 hole) reactionless mixture

Calculate IC by each chemical compound in the logarithm regression analysis that the dilution series of 10 μ M 3 or 10 times (usually since) percentage ratio of 4 concentration suppresses ₅₀Value.

In each experiment, the actual inhibition of reference compound is used for IC ₅₀Value is normalized to the benchmark with reference to the inhibitor meansigma methods:

Normalized IC ₅₀The IC of=mensuration ₅₀Reference compound IC ₅₀The reference compound IC of meansigma methods/mensuration ₅₀

For example: be 0.4 μ M with reference to inhibitor in the experiment, average out to 0.3 μ M

Test-compound is 1.0 μ M in the experiment, is normalized to: 0.3/0.4=0.75 μ M.

For example, D-82041 DEISENHOFEN or synthetic staurosporine derivatives are as reference compound.

Use this scheme, the chemical compound of discoverable type (I) suppresses to show the IC of 0.005-100 μ M scope to RET ₅₀Be worth, preferably show the IC of 0.01-2 μ M scope ₅₀

Can following proof chemical compound of the present invention as the efficient of the inhibitor of c-Abl protein-tyrosine kinase activity: as people such as Geissler, Cancer Res.1992; The described filtration binding assay of 52:4492-4498 carries out the vitro enzyme algoscopy in 96 orifice plates, it has following modification.Clone histidine-tagged c-Abl kinase domain also as people such as Bhat, J.Biol.Chem.1997; In baculovirus/Sf9 system, express described in the 272:16170-16175.By the cobalt metal chelating column is the protein of two step flow process purification 37kD (c-Abl kinases) of anion-exchange column then, and output is 1-2mg/L Sf9 cell people such as (, the list of references of quoting) Bhat.Identify that by SDS-PAGE the kinase whose purity of c-Abl is greater than 90% behind coomassie brilliant blue staining.This algoscopy contains (cumulative volume 30 μ L): c-Abl kinases (50ng), 20mM Tris HCI, pH 7.5,10mM MgCl ₂, 10 μ MNa ₃VO ₄, 1mM DTT and exist under the 1%DMSO condition with 30 μ g/mL poly--Ala, Glu, Lys, Tyr-6: the 0.06pCi/ of 2: 5: 1 (Poly-AEKY, Sigma P1152) measures [γ ³³P]-ATP (5 μ M ATP).

By adding 10 μ L 250mM EDTA cessation reactions and 30 μ L reactant mixtures being transferred in advance (MA USA), uses water rinse, uses 0.5%H then for Millipore, Bedford on the Immobilon-PVDF film that soaks 5 minutes in methanol ₃PO ₄Soaked 5 minutes and placed and take off the total chamber of the vacuum that is connected with vacuum source.Connect vacuum equipment on the point behind all samples also with 200 μ L 0.5%H ₃PO ₄Each hole of rinsing.Take off film and on shaking table, use 0.5%H ₃PO ₄Washing (4 times) and once with washing with alcohol.Count film after the drying at room temperature, place Packard Top Count 96 hole supports, and add the Microscint TM (Packard) in 10 μ L/ holes.

Use this detection system, the chemical compound of formula I suppresses to show the inhibition IC of 0.002-100 μ M scope to c-Abl ₅₀Be worth, show the inhibition IC of 0.002-5 μ M scope usually ₅₀Value.

Chemical compound of the present invention can followingly prove as the effect of KDR protein-tyrosine kinase activity inhibitor: the inhibition of the inductive receptor autophosphorylation of VEGF can be in cell, as verifying with another experiment in vitro in the Chinese hamster ovary celI of transfection, wherein said cell is expressing human VEGF-R2 receptor (KDR) constantly, is inoculated in the complete medium in the 6 porocyte culture plates (to contain 10% hyclone=FCS) and at 5%CO ₂Show about 80% converge in 37 ℃ of incubations up to it under the condition.Then with diluted chemical compound to be detected (no FCS contains 0.1% bovine serum albumin) and join in the cell in culture medium.(contrast comprises the culture medium of no chemical compound to be detected).37 ℃ of incubations added reorganization VEGF in two hours afterwards, and final VEGF concentration is 20ng/ml.37 ℃ of incubations after 5 minutes again are with twice of ice-cold PBS (phosphate buffer) washed cell and cracking in the lysis buffer in the every hole of 100 μ L immediately.Centrifugal then lysate is measured the protein concentration of supernatant to remove nucleus with commercial protein determination (BIORAD).Lysate can use or be stored in if necessary-20 ℃ immediately then.

Carry out sandwich enzyme-linked immunosorbent assay method and measure the VEGF-R2 phosphorylation: with the monoclonal antibody of VEGF-R2 (Mab 1495.12.14 for example; ProQinase, Freiburg Germany) is fixed in (available from the OptiPlateTM HTRF-96 of Packard) on the black ELISA flat board.Washing is dull and stereotyped then also contains Tween 20  (polyoxyethylene (20)-sorbitan monolaurate with being dissolved in, the 3%TopBlock  of phosphate buffer ICI/Uniquema) (PBST) (Juro, Cat#TB232010) saturated remaining free protein binding site.In these flat boards, spend the night then in 4 ℃ of incubated cell lysates (20 μ g protein/hole) with the link coupled anti-phosphotyrosine antibody of alkali phosphatase (PY20:AP is from Zymed).(washing is dull and stereotyped once more) (CDP-Star, instant contain Emerald II to use luminous AP substrate then; Applied Biosystems) proves combining of anti-phosphotyrosine antibody and the phosphorylation receptor of catching.In Packard Top Count MicroplateScintillation Counter, measure luminous.Difference between the signal of the signal of positive control (stimulating) and negative control (not stimulating) and the inductive VEGF-R2 phosphorylation of VEGF corresponding (=100%) with VEGF with VEGF.The percentage ratio that the activity of examined material is calculated as the inductive VEGF-R2 phosphorylation of VEGF suppresses, and wherein induces the maximum material concentration that suppresses of half to be defined as IC ₅₀(50% amount that suppresses).The chemical compound of formula I suppresses to show the IC of 0.005-20 μ M scope at this to KDR ₅₀, preferably show the IC between the 0.005-1 μ M ₅₀

Following mensuration Flt3 kinase inhibition: the recombinant baculovirus that produces the aminoacid 563-993 amino acid region of expressing human Flt-3 kytoplasm kinase domain with baculovirus donor carrier pFbacG01 (GIBCO).By from people c-DNA library (Clontech), the increase coded sequence in Flt-3 cytoplasmic structure territory of PCR.By making the dna fragmentation of amplification and pFbacG01 carrier be fit to coupled reaction with BamHI and HindIII digestion.The connection of these dna fragmentations produces baculovirus donor plasmids Flt-3 (1.1).Following the carrying out of purification of protein expression and gst fusion protein in the generation of virus, the sf9 cell:

The generation of virus: transfer vector (pFbacG01-Flt-3) transfection that will contain the Flt-3 kinase domain is laid on the selectivity agar plate in DH10Bac cell line (GIBCO) and with cells transfected.The bacterium colony that does not insert fusion sequence (being carried by antibacterial) in viral genome is for blue.Select single white colony also to pass through plasmid purification flow process isolated viral DNA (rod granule) from antibacterial of standard.In shaking bottle, use Cellfectin reagent viral DNA transfection Sf9 or Sf21 cell (American type culture collection) then.

Determining of Sf9 cell middle and small scale protein expression: from the cells transfected culture, collect the culture medium that contains virus and be used for infection to increase its titre.The culture medium that contains virus that obtains after two-wheeled infects is used for large-scale protein expression.For large-scale protein expression, with 5 * 10 ⁷Individual cell/plating 100cm ²Circular tissue culturing plate and infect with the culture medium (approximately 5MOI) that 1mL contains virus.It is centrifugal 5 minutes to scrape cell and 500 rev/mins from flat board after 3 days.Will be from 10-20 100cm ²Dull and stereotyped cell precipitation is resuspended in (25mMTris-HCI, pH7.5,2mM EDTA, 1%NP-40,1mM DTT, 1mM PMSF) in the ice-cold lysis buffer of 50mL.Stir on ice cell 15 minutes then 5000 rev/mins centrifugal 20 minutes.

The purification of GST label protein matter: will be added to (Pharmacia) and usefulness 10mL 25mM Tris-HCI on the 2mL glutathione agarose post through centrifugal cell lysate, pH 7.5,2mMEDTA, 1mM DTT, 200mM NaCl washing 3 times.Use 25mM Tris-HCI by 10 times (each 1mL) then, pH 7.5, the 10mM reduced glutathione, and 100mM NaCl, 1mMDTT, the protein of 10% glycerol eluting GST label also is stored in-70 ℃.

The measurement of enzymatic activity: the tyrosine protein kinase of carrying out purification GST-Flt-3 in 30 μ l final volume is measured, and contains 200-1800ng zymoprotein (depending on specific activity) in the wherein said 30 μ l final volume, 20mM Tris-HCl, and pH 7.6,3mM MnCl ₂, 3mM MgCl ₂, 1mM DTT, 10 μ M Na ₃VO ₄, 3 μ g/mL gather (Glu, Tyr) 4: 1,1%DMSO, 8.0 μ M ATP and 0.1 μ Ci[γ ³³P] ATP.Exist or do not exist under the inhibitor condition by measuring [γ ³³P] ATP ³³P is incorporated into poly-(Glu, Tyr) mensuration activity in the substrate.In 96 orifice plates, measuring (30 μ L) in 20 minutes and passing through to add 20 μ l 125mM EDTA termination under the condition as described below in room temperature.Subsequently 40 μ L reactant mixtures are transferred in advance with (MA USA), uses 0.5%H then with water rinse for Millipore, Bedford on 5 minutes the Immobilon-PVDF film of methanol immersion ₃PO ₄Soaked 5 minutes and placed and take off the total chamber of the vacuum that is connected with vacuum source.Connect vacuum equipment on the point behind all sample and with 200 μ L 0.5%H ₃PO ₄Each hole of rinsing.Remove film and on shaking table, use 1.0%H ₃PO ₄Wash 4 times, once with washing with alcohol.Count film after the drying at room temperature and place Packard Top Count96 hole support, add the Microscint TM (Packard) in 10 μ L/ holes.IC is calculated in the linear regression analysis that the percentage ratio of each chemical compound by bipartite four concentration (being generally 0.01,0.1,1 and 10 μ M) suppresses ₅₀Value.A protein kinase activity unit definition is 37 ℃ of 1 nanomole ³³P ATP is from [γ ³³P] ATP transfers to every milligram of protein of per minute of substrate protein white matter.The chemical compound of formula I shows the IC of 0.01-100 μ M scope to Flt-3 ₅₀Inhibiting value preferably shows the IC of 0.05-10 μ M ₅₀Inhibiting value.The chemical compound of formula I also suppresses other tyrosine protein kinase, and as especially c-Src kinases, c-Kit, VEGF-R and/or FGFR, it all participates in animal, and especially mammalian cell comprises the growth regulating and the conversion of people's cell.Suitable algoscopy is described among the Cancer Res.52,5353-8 (1992) people such as Andrejauskas-Buchdunger.Use this detection system, the chemical compound of formula I suppresses to show the IC of 0.005-100 μ M scope to c-Src ₅₀, show the IC of 0.005-5 μ M scope usually ₅₀The chemical compound of formula I also suppresses to show the IC of 0.005-10 μ M scope to c-kit ₅₀, show the IC of 0.005-5 μ M scope usually ₅₀, and FGFR-1 is suppressed at 10 μ M shows inhibition up to 95%.

The inhibition of IGF-1R and Ins-R can followingly be measured: use baculovirus donor carrier pfbgx3lGFIRcd to produce the recombinant baculovirus of the amino acid region 950-1337 in expressing human IGF-IR mature peptide cytoplasmic structure territory.Use pC5hinsR to produce the cDNA fragment of the amino acid region 919-1343 of kinase domain in the coding insulin human recipient cytoplasm.The Bac-to-Bac that uses recombinant baculovirus to produce ^TMThe clone of system (GIBCO BRL), expressing human IGF-IR and the fragment of Ins-R and glutathione-S-transferase (GST) fusion rotein that small scale purification factor Xa-can cut.From the cells transfected culture, collect the culture medium that contains virus and be used for infection to increase its titre.The culture medium that contains virus that two-wheeled infects the back acquisition is used for large-scale protein expression.The preparation cell extract also is added on glutathione agarose (Pharmacia) post.After the washing, then with the buffer solution elution GST label protein matter that contains glutathion.The protein of purification is stored in-70 ℃ the elution buffer.Containing 20mM Tris-HCl, pH 7.6,10mM MgCl ₂, 0.01mM Na ₃VO ₄, 1%DMSO, 1mM DTT, 3 μ g/ml poly (Glu, Tyr) 4: 1 and 10 μ M ATP (γ-[ ³³P]-ATP0.1 μ Ci) 30 μ l final volume in carry out the tyrosine protein kinase algoscopy of purification GST-IGF-1R and GST-lns-R.Room temperature was measured in 96 orifice plates and was stopped by adding 25 μ l 0.05MEDTA pH 7.0 in 20 minutes.With the liquid measure of 40 μ l parts with multichannel sample applicator point sample to the Whatman P81 film that places Millipore Microtiter filter house steward that is connected to the coarse vacuum source.After removing liquid, film is transferred to a series of 4 contains 0.5%H ₃PO ₄Bathing and an EtOH bathing (each shakes and hatches 10 minutes), drying, place Hewlett Packard house steward TopCount that added 10 μ l Microscint  and counting.The chemical compound of formula I is 10, and 000nM shows Ins-R and preferably to show the inhibition of 60-90% up to 90% inhibition.

The inhibition of Tek can followingly be measured: people such as Fabbro, Pharmacol.Ther.82 (2-3) 293-301 (1999) have described these kinase whose flow processs of expression, purification and mensuration.In brief, cut out glutathione S-transferase (GST) gene and insert the carrier of setting up 5530bp the cloning site of Fast-Bac baculovirus vector (GIBCO) from pAcG1 carrier (Pharmingen) with EcoRV and EcoRI with the N end cloning site that derives from pAcG1 fusion vector (FBG0).C end cloning site can be arbitrary cloning site (deriving from the Fast-Bac carrier) in used N end cloning site downstream.From ProQinase, Freiburg, Germany obtain (pAcG1, Pharmingen) KDR, Flt-1, Flk-1, Tek and the PDGFR-beta kinase domain that N end GST merges.The FBG1 that cuts out and be connected into EcoRI digestion by EcoRI is cloned into (FBG1-Tek) in the FBG1 carrier once more with Tek.The coded sequence in the whole cytoplasmic structures territory (aminoacid 538-972) by the whole cytoplasmic structures of pcr amplification c-Kit territory (aminoacid 544-976) and c-Fms from people uterus and people's bone marrow cDNA library (Clontech) respectively.Insert fragment cloning by dna fragmentation as BamHI-EcoRI and in FBG1, merge, produce FBG1-c-Kit and FBG1-c-Fms with GST with amplification.Cut out and be connected to by EcoRI among the FBG0 of EcoRI digestion Tek is cloned into (FBG-Tie2/Tek) in the FBG0 transfer vector once more.Obtain FGFR-1 and c-met kinase domain by PCR from people A431 cell.N end primer contains outstanding EcoRI site, and C end primer contains the XhoI site and is cloned in the transfer vector with auxiliary.After PCR fragment and FBG0 digest gel-purified cleaved products and connecting together form the kinases construct (FBG-Met, FBG-FGFR-1).

Each the kinase whose virus of scheme constructs that provides according to GIBCO.In brief, the transfer vector transfection that will contain kinase domain is laid on the agar plate of Blue-Gal, the IPTG, kanamycin, tetracycline and the gentamycin that contain recommended density in DH10Bac cell line (GIBCO).The bacterium colony that does not insert fusion sequence (being carried by antibacterial) in viral genome is for blue.Usually choose plasmid a small amount of preparation flow isolated viral DNA (rod granule) from antibacterial that single white colony also passes through standard.Use the scheme that provides with Bac-to-Bac test kit (GIBCO) at 25cm then ²Shake in the bottle with viral DNA transfection Sf9 cell or High Five cell (GIBCO).From the cells transfected culture, collect the culture medium that contains virus and be used for infection to increase its titre.The culture medium that contains virus that obtains after two-wheeled infects is used for large-scale protein expression.For large-scale protein expression, with 5 * 10 ⁷Individual cell/plating 100cm ²Circular tissue culturing plate and infect with the culture medium (approximately 5MOI) that 1mL contains virus.It is centrifugal 5 minutes to scrape cell and 500 rev/mins from flat board after 3 days.

Will be from 10-20 100cm ²Dull and stereotyped cell precipitation is resuspended in (25mM Tris-HCI, pH7.5,2mM EDTA, 1% NP-40,1mM DTT, 1mM PMSF) in the ice-cold lysis buffer of 50mL.Stir on ice cell 15 minutes then 5000 rev/mins centrifugal 20 minutes.Supernatant is added to the 25mM Tris-HCl that also uses 10ml on the 2ml glutathione agarose post, and pH 7.5,2mM EDTA, 1mM DTT, 200mM NaCl washing three times.Use 25mM Tris-HCl by 10 times (each 1ml) then, pH 7.5, the 10mM reduced glutathione, and 100mM NaCl, 1mM DTT, 10% glycerol eluting GST label protein matter also is stored in-70 ℃.

Algoscopy (30 μ l) contains the zymoprotein (depending on specific activity) of 200-1800ng, 20mMTris-HCl, and pH 7.6,3mM MnCl ₂, 3mM MgCl ₂, 1mM DTT, 10 μ MNa ₃VO ₄, 3 μ g/ml poly-(Glu, Tyr) 4: 1,8 μ M ATP (γ-[ ³³P]-ATP 0.1 μ Ci).The incubated at room reaction passed through to add 25 μ l 0.25M EDTA (pH 7.0) stopped reaction in 20 minutes then.With the liquid measure of 40 μ l parts with multichannel sample applicator point sample to the Whatman P81 film that places MilliporeMicrotiter filter house steward that is connected to the coarse vacuum source.After removing liquid, film is transferred to a series of 4 contains 0.5%H ₃PO ₄Bathing and an EtOH bathing (each shakes and hatches 10 minutes), drying, place Hewlett Packard house steward TopCount that added 10 μ l Microscint  and counting.By linear regression analysis calculating formula (I) chemical compound Tek is suppressed to show about 0.1-100 μ M IC ₅₀Value.

The inhibition of Cdk1 can followingly be measured: Cdk1/cycB:Cdk1/cycB is from ProQinase, Freiburg, and Germany obtains.Induce the sea star egg blast cell to enter M phase of cell cycle with 10 μ M 1-methyladenines, freezing and be stored in-80 ℃ in liquid nitrogen.When needing, also centrifugal as described homogenate oocyte such as (Arion people, people such as Cell 55:371-378 (1988) and Rialet, Anticancer Res.11:1581-1590 (1991)).At p9 ^CKShsPurification Cdk1/cycB kinases and as described recombined human p9 that uses such as (Azzi people, Eur.J.Biochem.203:353-360. (1992)) on-the sepharose 4B ^CKShsEluting.In brief, continue to derive from 4 ℃ of balances under the rotating condition supernatant and the p9 of oocyte ^CKShs-agarose granule 30 minutes.The thorough washing pearl is also used the activated cdk1/cycB kinases of p9CKShs (3mg/ml) eluting of purification.As (people such as Meijer, EMBO are J.1989 for people such as Arion, Cell 55:371-378 (1988); People such as 8:2275-2282 and Meijer, EMBOJ.1991; 8:2275-2282) described measurement Cdk1/cycB activity.The algoscopy of in 96 orifice plates, revising a little in room temperature 20 minutes.The Cdk1/cycB that contains 0.1-0.3U in the final volume of 30 μ l, 1mg/ml histone h1 be as substrate, 60mM β-phosphoglycerol, 30mM nitrobenzophenone phosphate ester, 25mM MOPS, 5mM EGTA, 15mM MgCl ₂, 1mM DTT, 0.1mMNa ₃VO ₄, 15 μ M ATP and 0.1 μ Ci γ- ³³P-ATP (75 μ M, 8800cpm/pmole).By adding 25 μ l 0.05M EDTA pH, 7.0 cessation reactions.With the liquid measure of 40 μ l parts with multichannel sample applicator point sample to the Whatman P81 film that places Millipore Microtiter filter house steward that is connected to the coarse vacuum source.After removing liquid, film is transferred to a series of 4 contains 0.5%H ₃PO ₄Bathing and an EtOH bathing (each shakes and hatches 10 minutes), drying, place Hewlett Packard house steward TopCount that added 10 μ l Microscint  and counting.The chemical compound of formula (I) is 10, and 000nM shows that the Cdk1 up to 100% suppresses.

The inhibition of c-Raf-1 can followingly be measured: by using the proteinic generation of triple infection Sf21 cell acquisition reorganization c-Raf-1 together of GST-c-Raf-1 recombinant baculovirus and v-Src and v-Ras recombinant baculovirus, that wherein said v-Src and v-Ras recombinant baculovirus are that active c-Raf-1 kinases produces is needed (people such as Williams, PNAS 1992; 89:2922-2926).Recruiting c-Raf-1 needs activatory Ras (v-Ras) and phosphorylation c-Raf-1 to need v-Src to cell membrane, with abundant activation it (people such as Williams, PNAS 1992; 89:2922-2926).Each 150mm plating 2.5 * 10 ⁷Individual cell also allows it to adhere to 1 hour in room temperature and 150mm plate.Culture medium (SF900II that contains 10%FBS) is bled and added recombinant baculovirus GST-C-Raf-1, v-Ras and v-Src to be respectively 3.0,2.5 and 2.5 MOI in the cumulative volume of 4-5mL.The incubated at room cell added the 15mL culture medium in 1 hour then.27 ℃ of cell 48-72hr of hatching infection.Scrape the Sf21 cell of infection, collect in the 50mL centrifuge tube and in the Sorvall centrifuge in 4 ℃ with 1100g centrifugal 10 minutes.With ice-cold PBS washed cell precipitation once and per 2.5 * 10 ⁷The 0.6mL lysis buffer cracking of individual cell.Blow and beat frequently on ice, realize the complete cracking of cell after 10 minutes.With the SS-34 rotor in the Sorvall centrifuge in 4 ℃ with 14,500g centrifuge cell lysate 10 minutes is also transferred to supernatant in the new pipe and is stored in-80 ℃.To per 2.5 * 10 ⁷Individual cell is with the equilibrated glutathione agarose 4B pearl purification c-Raf-1 from cell lysate that is filled with of the ice-cold PBS of 100uL.Allow GST-c-Raf-1 to shake at 4 ℃ and combine 1hr with pearl.To transfer in the pillar with the bonded GST-c-Raf-1 of pearl.Pillar washs once also with ice-cold Tris buffer salt solution washed twice with lysis buffer.Add ice-cold elution buffer and stop pillar and flow to allow free glutathion to destroy the interaction of GST-c-Raf-1 and glutathione agarose pearl.(1mL) collects in the test tube of pre-cooling with fraction.Each test tube contains 10% glycerol (final concentration) to keep kinase activity in freeze-thaw cycle.Purified GST-c-Raf-1 kinase protein fraction is stored in-80 ℃.

I κ B is as the kinase whose substrate of c-Raf-1.I κ B is expressed as the protein of His label and (is cloned and be so kind as to give by doctor Eder in antibacterial; ABM, Novartis, Basel).The BL21LysS antibacterial that contains I κ B plasmid grows into OD in the LB culture medium ₆₀₀Be 0.6 to use IPTG (1mM final concentration) to induce it to express I κ B in 3 hours then in 37 ℃, (little most advanced and sophisticated boundary line is set at ultrasonic buffer [50mM Tris pH 8.0 by supersound process cracking antibacterial then, 1mM DTT, 1mM EDTA] in each minute three times) and 10, centrifugal 15 minutes of 000g.Supernatant mixed with ammonium sulfate obtain 30% final concentration.Shake mixture 15 minutes then 10, centrifugal 15 minutes of 000g in 4 ℃.Precipitation is resuspended in the binding buffer liquid that contains 10mM BSA (Novagen) then.This solution is used for Ni-agarose (Novagen) and washs according to the Novagen handbook.With elution buffer (0.4M imidazoles, 0.2M NaCl, 8mM Tris pH 7.9) eluting I κ B from the pillar.At 50mM Tris pH8, dialysis contains proteinic fraction among the 1mM DTT.

Under the condition that has or do not exist inhibitor, pass through to measure ³³P is from [γ ³³P] ATP is incorporated among the IB activity of measuring the c-Raf-1 protein kinase.In 96 orifice plates, carried out this algoscopy in 60 minutes in room temperature.It contains (final volume of 30 μ l): c-rafl1 kinases (400ng), 25mM TrisHCl, pH7.5,5mM MgCl ₂, 5mM MnCl ₂, 10 μ M Na ₃VO ₄, 1mM DTT and 0.3 μ Ci/ measure [γ ³³P]-ATP (10 μ M ATP), use 600ng IB and have 1%DMSO.By adding 10 μ L 250mM EDTA cessation reactions and 30 μ L reactant mixtures being transferred in advance with (MA USA), uses water rinse, uses 0.5%H then for Millipore, Bedford on 5 minutes the Immobilon-PVDF film of methanol immersion ₃PO ₄Soaked 5 minutes and placed and take off the total chamber of the vacuum that is connected with vacuum source.On the point after all sample, connect vacuum and with 200 μ L 0.5%H ₃PO ₄Each hole of rinsing.Move down film and on shaking table, use 0.5%H ₃PO ₄Wash 4 times, once with washing with alcohol.Drying at room temperature places Packard TopCount 96 hole supports and adds MicroscintTM (Packard) the back counting film in 10 μ L/ holes.

The chemical compound of formula (I) shows in 0.1-10 μ M scope that preferably c-Raf-1 suppresses in 0.1-50 μ M scope.

The experiment of body internal evidence Ming Dynasty style (I) antitumor activity of compound: for example, take place in order to test the blood vessel whether formula (I) chemical compound that provides such as embodiment 1 hereinafter suppress the VEGF mediation in the body, tested the influence of described chemical compound to being taken place by the inductive blood vessel of the VEGF in the somatomedin transplant model in mice: the 0.8%W/V that packs in porous polytetrafluoroethylene cell comprises heparin (20 units per ml) agar of (containing or do not contain somatomedin (2pg/ml people VEGF)), then with its subcutaneous implantation C57/C6 mouse back side.Began the same day with described chemical compound to be tested (for example once a day 25,50 or 100mg/kg p.o.) or the above-mentioned mice of vehicle treated from implanting this cell, and continue four days.When processing finishes, put to death mice, and remove cell.Shift out carefully and be grown in cell blood vessel tissue on every side, it is weighed, and evaluate blood content (Drabkins method by the content of hemoglobin of measuring this tissue; Sigma, Deisenhofen, Germany).These are grown in cell tissue on every side presents (being characterized as on the histology comprises fibroblast and little blood vessel) dose dependent on weight and blood content growth previous verified these growth factor-induced, and verified described reaction (is seen people such as Wood JM by the antibody blocking of special neutralize VEGF, Cancer Res.60 (8), 2178-2189, (2000) and people such as Schlaeppi, J.Cacner Res.Clin.Oncol.125,336-342, (1999)).Utilize this model, can confirm the inhibitory action of formula (I) chemical compound.

Synthetic method

The chemical compound of formula (I) is by being similar to Alicade, E; De Mendoza, J; Garcia-Marquina, JM; Almera, C; J.Heterocycl.Chem.11,423 (1974) described methods prepare and get:

(a) in organic solvent with nitrile A-CH ₂-C ≡ N and Ethyl formate reaction generate the 3-oxo-propionitrile that replaces,

(b) in organic solvent the 3-oxo-propionitrile of the replacement of condensation step (a) and a hydrazine hydrate to form the 2H-pyrazole-3-yl amine of formula (III):

(d) nitrile that formylated replaces under the condition that has ethylate and Ethyl formate is with the 3-oxo-propionitrile of preparation formula (II):

(c) at the 2H-pyrazole-3-yl amine of 3-oxo-propionitrile that has shrinking type under the condition of organic solvent (II) and formula (III) to form the chemical compound of formula (I).

Particularly, the chemical compound of formula (I) prepares (Fig. 2) by condensation 3-oxo-propionitrile (II) in alcohol hydrochloric acid (ethanolic HCl) and corresponding 2H-pyrazole-3-yl amine (III).Above-mentioned 2H-pyrazole-3-yl amine (III) is by condensation one hydrazine hydrate and be dissolved in corresponding 3-oxo-propionitrile of organic solvent such as ethanol, two  alkane or AcOH and a few hours be prepared from high temperature (preferred 100 ℃) heating.The method for optimizing of preparation title compound pyrazoles part is to stir a hydrazine hydrate and corresponding 3-oxo-propionitrile 2-3 hour under 100 ℃ in acetic acid, adds aqueous hydrochloric acid then, and further reaction mixture refluxed 20 minutes.When R1 is not H, then use the hydrazine of corresponding replacement.Above-mentioned 3-oxo-propionitrile (I) and (II) undertaken by the Sodium ethylate and the Ethyl formate (in ethanol, refluxing hour) of corresponding nitrile by using prepared fresh that classical formylation reaction is synthetic to be obtained.Alternatively, also can use corresponding 3,3-dialkoxy-propionitrile (is similar to Seneci, P., Nicola, M., Inglesi, M., Vanotti, E., Resnati, G.Synth.Commun.29 (2), the described method of 311-341 (1999)) or 3-dimethylamino-acrylonitrile replace 3-oxo-propionitrile and carry out condensation reaction.

Fig. 2.

Alternatively, the chemical compound of formula (I) can be prepared by synthesizing pyrazolo [1,5-a] the pyrimidin-7-yl amine core support that carries corresponding functional group (X sees Fig. 3) earlier, wherein residue A, R ₂Or R ₃Can introduce by the known response that shows as Fig. 3 respectively.

Fig. 3

R wherein ₁, R ₂, R ₃With the definition of X, and when needs, afterwards obtainable formula (I) chemical compound is converted into different formula (I) chemical compound at the reaction (a) and (b) or (c) suc as formula (I) chemical compound; The salt of obtainable formula (I) chemical compound is converted into free chemical compound or different salt, perhaps obtainable free formula (I) chemical compound is converted into salt; And/or the isomer mixture of obtainable formula (I) chemical compound is separated into independent isomer;

Wherein for respond, the functional group's (if necessary) that does not participate in the raw material reacting exists with protected form by the blocking group of removing easily, and any subsequently blocking group all will be removed.

The preferred following reaction condition of difference:

Within this paper category, unless indicate in addition in the literary composition, what only be easy to remove is not that the group of formula (I) specific purpose end-product composition is called " blocking group ".The protective group effect of this type of blocking group; blocking group self and their cleavage reaction are existing to be described; for example such as J.F.W.McOmie; " blocking group in the organic chemistry "; Plenum Press; London and New York1973; T.W.Greene and P.G.M.Wuts; " blocking group in the organic synthesis "; the third edition; Wiley; New York 1999; " peptide " the 3rd volume (editor: E.Gross and J.Meienhofer); Academic Press; London and New York 1981; " Methoden der organischenChemie " (organic chemistry method); Houben Weyl; the 4th edition; Volume15/1; GeorgThieme Verlag; Stuttgart 1974; H.-D.Jakubke and H.Jeschkeit; " aminosauren; Peptide; Proteine " (aminoacid; peptide; protein); Verlag Chemie; Weinheim; Deerfield Beach and Basel 1982 and Jochen Lehmann; " Chemieder Kohlenhydrate:Monosaccharide und Derivate " (carbohydrate chemistry: monosaccharide and derivant thereof); Georg Thieme Verlag is in the canonical reference work of Stuttgart 1974.Blocking group be characterised in that can by solvolysis for example, reduction, photodecomposition or alternatively under physiological condition (for example by zymolysis) remove (undesirable supervention reaction does not promptly take place) easily.

The salt that can prepare formula (I) chemical compound by known mode itself with at least one salt forming group.For example, metallic compound that can be by using alkali metal salt (for example sodium salt of 2 ethyl hexanoic acid) such as suitable organic carboxyl acid, such as the organic alkali metal of corresponding hydroxide, carbonate or bicarbonate such as sodium hydroxide or potassium, carbonate or bicarbonate or alkaline earth metal compound, also can with corresponding calcium compounds or with ammonia or suitable organic amine handle this chemical compound, preferred chemistry amount or the only a small amount of superfluous salt forming agent of using prepares formula (I) the chemical compound salt with acidic-group.The acid-addition salts of formula (I) chemical compound is to obtain such as the usual manner with sour or suitable anion exchange agent treated.Can pass through for example to handle, thereby the mode that will be neutralized to isoelectric point, IP such as the salt of acid-addition salts forms formula (I) the chemical compound inner salt that comprises acid and alkaline salt forming group (for example free carboxylic group and free amino group) with weak base or with ion-exchanger.

Can change salt into free chemical compound with conventional method; For example, slaine and ammonium salt can be by changing with suitable acid treatment, and acid-addition salts can change by for example handling with suitable alkaline reagent.

Mixture of isomers can be obtained according to the present invention, and independent isomer can be in a manner known way it be separated into; For example can separate diastereomer by separating heterogeneity solvent mixture, recrystallization and/or chromatography (for example carrying out for example medium pressure liquid chromatography) through silica gel or through reverse post; In addition, racemate also can separate, for example by forming salt with optically pure salt-forming reagent, and then by such as fractional crystallization or by carrying out chromatography through the optically-active column material, thus the non-enantiomer mixture that separation can so obtain.

Can for example use chromatography, apportion design, (weight) crystallization process or the like according to standard method to intermediate with end product is processed and/or purification.

General procedure condition

Generally speaking, all programs that following condition is applied to above and hereinafter relates to, but preferred above or the reaction condition that hereinafter is particularly related to:

All said procedure steps can be carried out under known reaction condition own, the condition of preferred those special instructions, lacking or existing usually solvent or diluent, preferably agents useful for same is inertia but dissolves their solvent or diluent, can lack or have catalyst, condensing agent or nertralizer, for example cationite be (for example with H ⁺Form, this depends on the reaction and/or the character of reagent) ion-exchanger, can be low temperature, room temperature or high temperature, for example approximately-100 ℃ to about 190 ℃ temperature range, preferably approximately-80 ℃ to about 150 ℃ (for example-80 to-60 ℃, room temperature ,-20 to 40 ℃ or reflux temperature), at ambient pressure or in closed container (as required under pressure), and/or can be in noble gas, for example argon or nitrogen.

In all stages of reaction, the isomer mixture that forms can be separated into independent isomer such as diastereomer or enantiomer, perhaps be separated into the isomer mixture of any hope, the mixture of racemate or diastereomer for example for example is similar to the method described in " additional process steps ".

Unless in method is described, specify, otherwise the reagent that those reagent that are fit to any concrete reaction are selected from comprises the reagent of those special instructions, perhaps water for example, esters such as low alkyl group-lower alkanoic acid ester (for example ethyl acetate), ethers such as aliphatic ethers (for example Anaesthetie Ether) or ring-type ethers (for example oxolane or two  alkane), liquid aromatic hydrocarbon such as benzene or toluene, such as methanol, the alcohol of ethanol or 1-or 2-propanol, nitrile such as acetonitrile, halogenated hydrocarbons such as dichloromethane or chloroform, amide such as dimethyl formamide or dimethyl acetylamide, base such as heterocyclic nitrogen base (for example pyridine or N-methylpyrrolidin-2-ketone), carboxylic acid anhydrides such as lower alkane anhydride (for example acetic anhydride), such as cyclohexane extraction, the cyclic hydrocarbon of hexane or isopentane, straight-chain hydrocarbons and branched-chain hydrocarbons, the perhaps mixture of these solvents, for example aqueous solution.This type of solvent mixture also can be used for for example carrying out chromatography or separation.

Chemical compound comprises their salt, also can obtain with the form of hydrate, and perhaps their crystal can for example comprise the solvent that is used for crystallization.Can there be different crystal forms.

The present invention also relates to those chemical compounds that can obtain from this method any stage intermediate as raw material and carry out remaining step or wherein raw material forming under the reaction condition or be used or under reaction condition, generate chemical compound that the method according to this invention can obtain and it such as is further processed in position at method of form with the form of the derivant form of protected form or salt (for example with).In the method for the invention, preferably use those to be created in the raw material that preamble is described as especially valuable formula (I) noval chemical compound.Identical with reaction condition described in the embodiment or similar reaction condition is particularly preferred.

According to the preferred embodiment of the invention:

In following preferred embodiment, use above to replace general expression way, thereby produce the embodiment that the present invention is more preferably with corresponding more concrete definition provided below.

Preferred formula (I) chemical compound, the purposes of its tautomer or its officinal salt, tyrosine protein kinase dependence disease wherein to be treated are the proliferative diseases that depends on any one or multiple following tyrosine protein kinase: c-Abl especially, Bcr-Abl, c-Kit, c-Raf, Flt-1, Flt-3, KDR, Her-1, the PDGFR-kinases, c-Src, the RET-receptor kinase, FGF-R1, FGF-R2, FGF-R3, FGF-R4, liver is joined protein receptor kinases (EphB2 kinases for example, EphB4 kinases and relevant Eph kinases), casein kinase (CK-1, CK-2, G-CK), Pak, ALK, ZAP70, Jak1, Jak2, Axl, Cdk1, cdk4, cdk5, Met, FAK, Pyk2, Syk, insulin receptor kinase, Tie-2 or such as Bcr-Abl, c-Kit, cdk1, c-Raf, Flt-3, FGF-R3, the PDGF-receptor, the kinases constitutive activation mutant (activity kinases) of RET and Met.

More preferably, formula (I) chemical compound can be used for the treatment of and depends on following kinase whose proliferative disease: c-abl, Flt-3, KDR, c-Src, RET, EphB4, c-kit, cdk1, FGFR-1, c-raf, Her-1, Ins-R and Tek.

The invention particularly relates to formula (I) compound or pharmaceutically acceptable salt thereof,

Wherein:

R ₂Be H; That replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; That replace or unsubstituted aliphatic residue; Functional group; Perhaps the replacement by linking group or atom and the connection of pyrazolo [1,5a] pyrimidine-ring or unsubstituted aryl, replacement or unsubstituted heteroaryl or replacement or unsubstituted aliphatic residue;

R ₃Can be H, replacement or unsubstituted aryl, replacement or unsubstituted heteroaryl, replacement or unsubstituted aliphatic residue, functional group or can be by linking group or atom aliphatic residue with the connection of pyrazolo [1,5a] pyrimidine-ring,

R ₂Or R ₃In be one of at least that replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; Perhaps pass through replacement or unsubstituted heteroaryl or replacement or the unsubstituted aromatic yl residue that a linking group or atom and pyrazolo [1,5a] pyrimidine-ring connect;

A is H, halogen (as bromine), aliphatic series part, functional group, replacement or unsubstituted aryl or replacement or unsubstituted heteroaryl,

And R ₁Be H, halogen or low alkyl group,

And formula (I) chemical compound is in treatment kinases dependence disease or the purposes in the pharmaceutical preparation of preparation treatment kinases dependence disease.

The invention still further relates to the compound or pharmaceutically acceptable salt thereof of formula (I),

Wherein

R ₂Be H; That replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; That replace or unsubstituted aliphatic residue; Functional group; Perhaps the replacement by linking group or atom and the connection of pyrazolo [1,5a] pyrimidine-ring or unsubstituted aryl, replacement or unsubstituted heteroaryl or replacement or unsubstituted aliphatic residue;

R ₃Can be H, replacement or unsubstituted aryl, replacement or unsubstituted heteroaryl, replacement or unsubstituted aliphatic residue, functional group or can pass through linking group or atom and pyrazolo [1,5a] replacement or the unsubstituted aliphatic residue that connects of pyrimidine-ring

R ₂Or R ₃In be one of at least that replace or unsubstituted aryl; That replace or unsubstituted heteroaryl; Perhaps pass through replacement or unsubstituted heteroaryl or replacement or the unsubstituted aromatic yl residue that a linking group or atom and pyrazolo [1,5a] pyrimidine-ring connect; And condition is R ₂With A can not all be unsubstituted phenyl;

A be H, halogen (as bromine), aliphatic series part, functional group, replacement or unsubstituted aryl or heteroaryl,

And R ₁Be H, halogen or low alkyl group,

And formula (I) chemical compound is in treatment kinases dependence disease or the purposes in the pharmaceutical preparation of preparation treatment kinases dependence disease.

More preferably such formula (I) compound or pharmaceutically acceptable salt thereof, wherein

Connect atom or group and be selected from alkyl (as-CH ₂-), oxygen base-O-, ketone-CO-, sulfenyl-S-, sulfonyl-SO ₂-, sulfoxide-SO-, amine-NH-or-NR-, carboxylic acid, alcohol, ester (COO-), amide (--CONR-,-CONHR '-), sulfoamido (SO ₂NH-,-SO ₂NR '-), (SO ₃-), sulfoxide (SO-), amino, carbamide (NH-CO-NH-,-NR-CO-NH-,-NH-CO-NR-,-NR-CO-NR-), ether (O-), carbamate (NH-CO-O-,-NR-CO-O-) or inverse amide, sulfonamide and ester (NH-CO-,-NR-CO-,-NH-SO ₂-,-NR-SO ₂-,-OOC-); Especially preferred alkyl is (as-CH ₂-), oxygen base-O-, ketone-CO-, sulfonyl-SO ₂-, sulfoamido (SO ₂NH-,-SO ₂NR '-), (SO ₃-) and carbamide (NH-CO-NH-,-NR-CO-NH-,-NH-CO-NR-,-NR-CO-NR-),

And functional group is selected from carboxylic acid, hydroxyl, halogen, cyano group (CN), ether (OR), ketone (CO-R), ester (COOR), amide (CONH ₂,-CONHR ,-CONRR '), thioether (SR), sulfonamide (SO ₂NH ₂,-SO ₂NHR ,-SO ₂NRR '), sulfone (SO ₂-R), sulfoxide (SO-R), amine (NHR, NR ' R), carbamide (NH-CO-NH ₂,-NH-CO-NHR), ether (O-R), halogen, carbamate (NH-CO-OR), aldehyde functional group (CHO), also can be then inverse amide, sulfonamide and ester (NH-CO-R ,-NH-SO ₂-R ,-OOC-R); Especially preferred halogen, hydroxyl, ether (OR), amide (CONH ₂,-CONHR ,-CONRR '), sulfonamide (SO ₂NH ₂,-SO ₂NHR ,-SO ₂NRR '), (NHR, NR ' are R) and carbamide (NH-CO-NH for amine ₂,-NH-CO-NHR),

Perhaps they are particularly useful for the purposes of pharmaceutical compositions, perhaps are used to diagnose or treat homoiothermic animal, especially people's purposes.

Preferred especially such formula (I) compound or pharmaceutically acceptable salt thereof, wherein

A is H, halogen (for example Br) or aryl (for example phenyl or benzyl) or heterocyclic radical (for example pyridine radicals, indyl or benzothienyl),

Wherein said aryl or heterocyclic radical can be with being no more than 4 substituent groups that preferably are no more than 2 and replace or do not replace, wherein said substituent group can be identical or different, and be independently selected from: halogen (for example Cl or Br), hydroxyl, amino, amino low alkyl group (for example dimethylamino), amino lower alkoxy (for example amine ethoxylate), low alkyl group (for example methyl), lower alkoxy (for example methoxyl group), replacement or unsubstituted sulfoamido (for example benzene sulfinyl amido, chlorobenzene sulfonamide base or dichloro-benzenes sulfoamido), carbamate; R ₄R ₅, R wherein ₄And R ₅Can be the same or different, and be independently H, low alkyl group (for example methyl, ethyl or propyl group); Perhaps R ₄And R ₅And common 3-to the 8-unit heterocycle (for example piperazinyl or low alkyl group piperazinyl) that comprises 1-4 nitrogen, oxygen or sulphur atom that forms of N atom, wherein work as R ₄And R ₅And N is when forming heterocycle, and this heterocycle can be replaced wherein preferred piperazinyl, pyrrolidinyl, alkyl (for example methyl) or hydroxyalkyl (for example ethanyl) by any substituent group of describing in 1,2 or the more a plurality of literary composition.By R ₄And R ₅And the heterocyclic example that forms of N comprises that morpholinyl (can not be substituted or replace with methyl or dimethyl), piperazinyl (can not be substituted or replace with 1,2 or 3 substituent group, wherein said substituent group preferable methyl, oxygen base or ethanol) or piperadinyl (can not be substituted or replace the preferred pyrrolidinyl of wherein said substituent group, amine, alkylamine, methyl amine, dialkylamine, dimethyl amine or diethylamide with 1,2 or 3 substituent group);

R ₂Be H, C ₁-C ₃Low alkyl group (for example methyl) or aryl (for example phenyl or benzyl) or heterocyclic radical (for example pyridine radicals, indyl, thienyl, thiazolyl or benzothienyl), wherein said aryl or heterocyclic radical can be with being no more than 4 substituent groups that preferably are no more than 2 and replace or do not replace, wherein said substituent group can be identical or different, and be independently selected from: halogen (for example Cl, F or Br), hydroxyl, amino, amino low alkyl group, C ₁-C ₃Low alkyl group, alkoxyl (for example methoxyl group and benzyloxy, wherein said benzyl rings can be substituted or not be substituted, for example 3, the 4-dichloro-benzyloxy), sulfoamido, replacement or unsubstituted benzene sulfinyl amido (for example 2,3-dichloro-benzenes sulfoamido), replacement or unsubstituted sulfonate (for example chlorphenyl sulfonate), replacement or unsubstituted carbamide (for example 3-three fluoro-methyl-phenyl carbamide or 4-morpholine-4-base-3-trifluoromethyl-phenyl-carbamide) or carbamate (for example ethyl-N-carbanilate);

R ₃Be H, C ₁-C ₃Alkyl, methyl, phenyl, pyridine radicals or oxaz-5-base;

Perhaps they are particularly useful for the purposes of pharmaceutical compositions, perhaps are used to diagnose or treat homoiothermic animal, especially people's purposes.

Especially the purposes of preferred formula (I) compound or pharmaceutically acceptable salt thereof in the pharmaceutical preparation of preparation treatment kinases dependence disease.

Same preferred formula (I) compound or pharmaceutically acceptable salt thereof is as indicated above at treatment kinases dependence disease, especially depend on described kinase whose disease and (especially unusual high expressed or constitutive activation) described kinases dependence disease or depend on the disease that activates described kinase pathways or depend on the purposes of two or more described kinase whose diseases arbitrarily.

In more generalized understanding to the present invention, the kinases dependence disease can be a proliferative disease, comprise the hyperplasia disease, leukemia, hyperplasia, the fibrosis (fibrosis of lung especially for example, but the fibrosis that other type is also arranged is as renal fibrosis), the smooth muscle proliferation (for example narrowing down or restenosisization after the revascularization) in blood vessel generation, psoriasis, atherosclerosis and the blood vessel.

The method that very preferably comprises the formula of using (I) compounds for treating kinases dependence disease, wherein said disease to be treated is a proliferative disease, preferred optimum or malignant tumor especially, the more preferably brain cancer, renal carcinoma, hepatocarcinoma, adrenal carcinoma, bladder cancer, breast carcinoma, gastric cancer (especially gastric cancer), ovarian cancer, colon cancer, rectal cancer, carcinoma of prostate, cancer of pancreas, pulmonary carcinoma (especially SCLC), cancer of vagina, thyroid carcinoma, sarcoma, glioblastoma, multiple myeloma or human primary gastrointestinal cancers, especially the tumor of colon cancer or colorectal adenoma or cervical region and head, the perhaps hyperplasia of epidermis psoriasis especially, the prostate hyperplasia, neoplasia, especially the neoplasia of epithelial cell character, preferred mastocarcinoma or leukemia.Equally preferably use formula (I) compounds for treating atherosclerosis, thrombosis, psoriasis, scleroderma and fibrotic method.

The chemical compound of formula (I) can cause tumor regression, and stops the formation of neoplasm metastasis and the growth of neoplasm metastasis (comprising small transfer).In addition, also they can be used for the treatment of hyperplasia (for example psoriasis), prostate hyperplasia and neoplasia, the especially neoplasia of epithelium character (for example breast carcinoma) of epidermis.The chemical compound of formula (I) can also be used for the treatment of the disease of immune system that relates to several tyrosine protein kinase or especially relate to other tyrosine protein kinase scope of branch; In addition, the chemical compound of formula (I) also can be used for the treatment of the disease that the signal transmission relates to a tyrosine protein kinase at least or especially relates to the maincenter or the peripheral nervous system of those tyrosine protein kinase that are selected from the tyrosine protein kinase of mentioning especially.

Therefore, the KDR inhibitor is particularly suitable for treating the disease that relates to the vegf receptor tyrosine kinase overexpression.In these diseases, especially retinopathy, senile degeneration of macula, psoriasis, hemangioblastoma, hemangioma, arteriosclerosis, such as rheumatoid or rheumatic inflammatory disease, especially the inflammatory diseases of arthritis (for example rheumatoid arthritis) or other chronic inflammatory disease (for example breathing heavily for a long time), tremulous pulse or transplanting artery are atherosis, endometriosis and especially neoplasia disease, for example so-called solid tumor (gastrointestinal tract especially, pancreas, breast, stomach, cervix uteri, bladder, kidney, prostate, ovary, endometrium, lung, the cancer of brain, melanoma, kaposi's sarcoma, the squamous cell carcinoma of head and cervical region, malignant pleural mesothelioma, lymphoma or multiple myeloma) and liquid tumor (for example leukemia) be even more important.

Flt3 (FMD sample tyrosine kinase) specifically expressing in hemopoietic progenitor cell and in the CFU-GM of lymphocyte and myelocytic series.Confirmed that the FIt3 gene comprises AML (acute myelogenous leukemia), follows three to be among the leukemic adult of AML (AML/TMDS), ALL (acute lymphoblastic leukemia), CML (chronic granulocytic leukemia) and myelodysplastic syndrome of myelodysplasia and the child expression to be arranged all suffering from, therefore, preferably use these diseases of compounds for treating of formula (I).In patient AML of about 25%-30%, found the activity mutant of FIt3.Therefore, the evidence that Flt3 acts in human leukemia is constantly accumulation, and the chemical compound of useful pyrazolo [1,5-a] pyrimidin-7-yl-amine derivative, especially formula (I) is particularly useful in treatment the type disease as the Flt3 inhibitor and (sees people such as Tse, Leukemia according to the present invention 15(7), people such as 1001-1010 (2001), Tomoki, Cancer Chemother.Pharmacol. 48(Suppl.1), people such as S27-S30 (2001), Birkenkamp, Leukemia 15(12), people such as 1923-1921 (2001), Kelly, Neoplasia 99(1), 310-318 (2002)).

In chronic granulocytic leukemia (CML) since hematopoietic stem cell (HSC) thus in mutual equilibrated chromosome translocation produced the BCR-ABL fusion gene.The Bcr-Abl fused protein that the latter encodes carcinogenic.The ABL coding is subjected to the protein tyrosine kinase of strict regulation and control, it plays a major role in regulating cell proliferation, adhesion and apoptosis, and the activated kinases of BCR-ABL fusion gene code set molding, the zymogenesis HSC of this constitutive activation, thereby produce and to show as the clonal proliferation that is not subjected to regulate, to the phenotype that the adhesive capacity of bone marrow matrix descends and the apoptotic response that mutation stimulates is reduced, these phenotypes make it gradually to vicious transformation.Make that finally granulocyte can not the bud into mature lymphocyte, and discharge into circulation, and then cause the mature cell deficiency, and cause the sensitivity that infects is increased.Existing description shows that the ATP competitive inhibitor of Bcr-Abl stops the path (for example P-3 kinases and STAT5) of this kinase activator mitogenesis and anti-apoptotic, thereby causes BCR-ABL phenotype cell death, and therefore effective therapy of anti-CML is provided.Therefore, useful pyrazolo [1,5-a] pyrimidin-7-yl-amine derivative, especially formula (I) chemical compound according to the present invention especially is fit to the disease that treatment relates to its overexpression as the Bcr-Abl inhibitor, especially such as the leukemia of CML or ALL.

Therefore, formula (I) chemical compound that suppresses the tyrosine kinase of EGF-R or the protein hydroxyphenylaminopropionic acid kinase activity that other is mentioned for example treat optimum or malignant tumor in be useful.For example formula (I) chemical compound is except the vascularization of the solid tumor that can suppress to be triggered by VEGF, can also suppress simultaneously to have not to be subjected to EGF-R and/or the active tumor growth of ErbB-2 regulated.The active antitumous effect (seeing WO02/41882) that improves that caused of this associating.In addition, the use of this double inhibitor has reduced the risk of drug-drug interactions, and compares the further total amount of application that has reduced medicine with conjoint therapy.Can slow down tumor growth or realize tumour regression of formula (I) chemical compound, and can suppress that neoplasm metastasis forms and the micrometastasis growth.They especially can be used for the case (psoriasis) of epidermis hyperplasia, be used for the treatment of picture such as nonsmall-cell lung cancer, squamous cell carcinoma (head and cervical region), breast carcinoma, gastric cancer, ovarian cancer, colon cancer and carcinoma of prostate and gliomatous solid tumor, and be used for the treatment of in the leukemia of especially for example acute myeloid leukemia (AML) and chronic myeloid leukemia (CML).In addition, the chemical compound of formula (I) can be used in the treatment those relate to some protein tyrosine kinase and/or (in addition) serine/threonine protein matter kinases or especially relate to independent protein tyrosine kinase and/or the kinase whose disease of immune system of (in addition) serine/threonine protein matter; The chemical compound of formula (I) also can be used in those signal transmission of treatment, and to relate to several protein tyrosine kinases and/or (in addition) serine/threonine protein matter kinase whose or relate to the disease of especially single protein tyrosine kinase and/or kinase whose central nervous system of (in addition) serine/threonine protein matter or peripheral nervous system.

Think that blood vessel is the absolute essential condition of tumor that those growths surpass about 1-2mm maximum gauge; When arriving this limit, oxygen and nutrient can provide to tumor cell by diffusion.Therefore, each tumor is not considered its origin and reason thereof, and its growth just depends on the blood vessel generation after arriving a certain size.

Have three kinds of main mechanism in the anti-tumor activity of angiogenesis inhibitor, to play an important role: 1) suppress blood vessel especially blood capillary grow into the immobilized tumor of blood vessel, just make tumor not grow only because apoptosis and propagation reach balance like this; 2) thus owing to there is not blood flow turnover tumor to stop the migration of tumor cell; And 3) inhibition of endothelial cell proliferation, thus avoided being applied to paracrine growth stimulation effect in the peripheral organization by endotheliocyte (being arranged on the blood vessel usually).

The present invention also can be used to prevent or treat the disease that initiation is taken place by the persistence blood vessel, as psoriasis; Kaposi's sarcoma; Restenosis, for example restenosis of stent-induced; Endometriosis; Crohn disease; Hodgkin; Leukemia; Arthritis is as rheumatoid arthritis; Hemangioma; Fibrohemangioma; Ophthalmic is as diabetic retinopathy and neovascular glaucoma; Kidney disease is as glomerulonephritis; Diabetic nephropathy; Malignant nephrosclerosis; The thrombotic microangiopathy syndrome; Transplant rejection and glomerulopathy; Fibrotic disease is as the sclerosis of liver; The mesangial cell proliferation disease; Arteriosclerosis; Neural tissue injury.The present invention also can be used for suppressing the vascular reocclusion after the balloon catheter treatment, after being used for the blood vessel reparation or being used to insert the machinery such as support that keeps vessel open, as immunosuppressant,, be used for the treatment of senile plaque and contact dermatitis as assisting of no scar wound healing.

According to the present invention, formula (I) the compound or pharmaceutically acceptable salt thereof purposes of illustration in " embodiment " hereinafter most preferably.

Pharmaceutical composition

The present invention also relates to comprise the pharmaceutical composition of formula (I) chemical compound, relate to their purposes of therapeutic (aspect the present invention is more generalized, also relating to preventative) treatment or the method in the preferred disease of treatment kinases dependence disease, especially above narration, relate to the chemical compound of described purposes, and relate to the preparation of pharmaceutical preparation, especially described purposes pharmaceutical preparation.The invention still further relates to formula (I) the chemical compound prodrug that is converted into described formula (I) chemical compound in the body.Therefore, any formula of referring to (I) chemical compound also is interpreted as formula (I) chemical compound corresponding precursor medicine as required and advantageously.

Pharmaceutically acceptable chemical compound of the present invention also can be used for for example preparing the pharmaceutical composition that comprises effective dose formula (I) compound or pharmaceutically acceptable salt thereof, and described formula (I) compound or pharmaceutically acceptable salt thereof is as pharmaceutically suitable carrier or the mixing of active component with the inorganic or organic solid or the liquid of one or more significant quantities.

The invention still further relates to and be fit to homoiothermic animal especially people (perhaps from the cell or the cell line of homoiothermic animal especially people, as lymphocyte) use, be used for the treatment of or prevent (the present invention more generalized aspect) pharmaceutical composition disease, that comprise a certain amount of formula (I) compound or pharmaceutically acceptable salt thereof that the inhibition of kinase activity is reacted, described a certain amount of formula (I) compound or pharmaceutically acceptable salt thereof is effectively to described inhibition, especially when itself and at least a pharmaceutically suitable carrier one time-out.

According to pharmaceutical composition of the present invention for those through enteral (for example nose, rectum or the oral cavity) or parenteral (for example intramuscular or intravenous) be applied to homoiothermic animal (especially people), separately or with the pharmaceutical composition that comprises the effective dose pharmacy activity component of pharmaceutically suitable carrier of significant quantity.The dosage of effective ingredient depends on species, body weight, age and individual condition, the individual drugs dynamic metabolism data of homoiothermic animal, disease and mode of administration to be treated.

The present invention also relates to treat the treatment of diseases method that kinase whose inhibition is reacted, described method comprise to homoiothermic animal especially (for example because a kind of above-mentioned disease needs the people of this treatment) use the prevention of (anti-described disease) or treatment effective dose according to formula of the present invention (I) chemical compound.

Formula (I) compound or pharmaceutically acceptable salt thereof to homoiothermic animal for example approximately the dosage used of the people of 70kg body weight preferably from about 3 milligrams/people/sky to about 10 gram/people/sky, be more preferably from about 10 milligrams/people/sky extremely about 1.5 gram/people/sky, about 1000 milligrams/people/sky extremely most preferably from about 100 milligrams/people/sky, preferably be divided into the individual for example single dose of identical size of 1-3.Generally speaking, the child accepts half of adult's dosage.

This pharmaceutical composition comprises from about 1% to about 95%, preferred from about 20% to about 90% active component.According to pharmaceutical composition of the present invention can be for example form of unit dose, for example ampulla, bottle, suppository, lozenge, tablet or Capsule form.

Pharmaceutical composition of the present invention is by known mode itself, and for example dissolving, lyophilizing, mixing, granulation or the forming process by routine is prepared.

The preferred solution that uses active component, also have suspension, particularly isotonic aqueous solution or suspension, for example for only comprising active component or comprising active component and during the freeze-dried composition of carrier (for example mannitol), described solution or suspension can preparations before using.This pharmaceutical composition can be sterilized, and/or comprise, and by known mode itself, for example be prepared by the dissolving of routine or the mode of freeze-drying process such as the salt of antiseptic, stabilizing agent, wetting agent and/or emulsifying agent, solubilizing agent, adjusting osmotic pressure and/or the excipient of buffer agent.Described solution or suspension can comprise the material that increases viscosity, for example Sodium Tvlose, carboxymethyl cellulose, glucosan, polyvidon or gelatin.

In the oil suspension comprise as oil component, be generally used for injecting plant, the synthetic or semi-synthetic oil of purpose.What especially can mention is such liquid aliphatic acid esters, its comprise as acid ingredient, have 8-22, especially a long-chain fatty acid of 12-22 carbon atom, such as lauric acid, tridecanoic acid, tetradecanoic acid, pentadecanoic acid, palmitic acid, heptadecanoic acid, stearic acid, arachidic acid, mountain Yu acid or corresponding unsaturation acid, for example oleic acid, elaidic acid, erucic acid, brasileic acid or linoleic acid, if desired, add antioxidant such as vitamin E, beta-carotene or 3,5-two-tertiary butyl-4-hydroxy toluene in the described liquid aliphatic acid esters.The pure composition maximum of those fatty acid esters has 6 carbon atoms, and is such as single, double or triatomic monohydroxy or polyhydroxy-alcohol, such as methanol, ethanol, propanol, butanols or amylalcohol or its isomer, but preferred ethylene glycol and glycerol.Therefore, mention the example of following fatty acid ester: ethyl oleate, isopropyl myristate, isopropyl palmitate, " LabrafilM 2375 " (polyoxyethylene triolein, Gattefosse, Paris), " Miglyol 812 " (triglyceride of the satisfied fatty acid of tool C8 to C12 chain length, Huis AG, Germany) but especially such as the vegetable oil of Oleum Gossypii semen, almond oil, olive oil, Oleum Ricini, Oleum sesami, soybean oil and preferred especially Oleum Arachidis hypogaeae semen.

Injectable composition prepares under aseptic condition with conventional method; Be applied to equally described compositions pack into ampoule or bottle and seal this container.

Can this mixture be processed as tablet, lozenge or capsule (if expectation or need) by combined activity composition and solid carrier, granulation gained mixture (if desired) and after adding suitable vehicle, thereby obtain oral pharmaceutical composition.Active component also they may be integrated into plastic carrier, so that can spread in the mode that quantity can be surveyed or discharge.

Specially suitable carrier has filler, lactose for example, sucrose, the sugar of mannitol or Sorbitol, cellulose preparation and/or such as the calcium phosphate of tricalcium phosphate or calcium hydrogen phosphate, and binding agent, for example use such as corn, Semen Tritici aestivi, the gelatinized corn starch of Oryza sativa L. or potato starch, gelatin, tragcanth, methylcellulose, hydroxypropyl emthylcellulose, Sodium Tvlose and/or polyvinylpyrrolidone, and/or disintegrating agent (if desired), for example above-mentioned starch, and/or carboxymethyl starch, crosslinked polyvinylpyrrolidone, agar, alginic acid or its salt such as sodium alginate.Excipient is flowing regulator and lubricant especially, for example silicic acid, Talcum, stearic acid or its salt and/or Polyethylene Glycol such as magnesium stearate or calcium stearate.The lozenge sheet heart with suitable, randomly the coating of intestinal provides, comprise arabic gum, Talcum, polyvinylpyrrolidone, Polyethylene Glycol and/or titanium dioxide, perhaps be used to prepare coating by the coating solution that is dissolved in suitable organic solvent, perhaps in order to prepare enteric coating, by for example ethyl cellulose phthalate or the hydroxypropylmethyl cellulose phthalate preparation of suitable cellulose preparation solution.Capsule is with the dry-packing capsule of gelatin preparation with gelatin and such as the sealing soft capsule of the plasticizer preparation of glycerol or Sorbitol.Described dry-packing capsule can comprise the active component of particle form, for example has filler such as lactose, such as the binding agent of starch and/or such as the fluidizer of Talcum or magnesium stearate, and also has stabilizing agent if desired.In soft capsule,, also can add stabilizing agent and/or antibacterial preferably with active component dissolving or be suspended in the suitable oiliness excipient such as fatty oil, paraffin oil or liquid macrogol.For example in order to differentiate purpose or, can in tablet or lozenge coating or capsule shell, to add dyestuff or pigment in order to indicate the various dose of active component.

Combination

Formula (I) chemical compound also can be used for and other antiproliferative combination.Described antiproliferative is including, but not limited to aromatase inhibitor; antiestrogen; the topoisomerase I inhibitor; the topoisomerase II inhibitor; microtubule active agent; alkylating agent; histone deacetylase inhibitors; the chemical compound of inducing cell atomization; cyclooxygenase-2 inhibitor; the MMP inhibitor; the mTOR inhibitor; the antineoplastic antimetabolite; platinum compounds; the chemical compound of targeting/reduction protein kinase or the active chemical compound of lipid kinase and further angiogenesis inhibitor; targeting; the chemical compound of reduction or Profilin matter phosphatase or lipid phosphatase activity; the GnRF agonist; antiandrogen; the methionine aminopeptidase inhibitor; diphosphate; biological response modifier; anti proliferative antibody; heparanase inhibitors; the carcinogenic isotype inhibitor of Ras; the telomerase inhibitor; proteasome inhibitor; the reagent that is used for the treatment of the blood malignant tumor; targeting; reduce or the active chemical compound of inhibition Flt-3; the Hsp90 inhibitor; temozolomide (TEMODAL ) and formyl tetrahydrofolic acid.

Term used herein " aromatase inhibitor " relates to and suppresses estrogen and generate and promptly suppress the chemical compound that substrate androstenedione and testosterone are separately converted to estrone and estradiol.This term is including, but not limited to steroid, especially atamestane, exemestane and formestane, and nonsteroidal especially, especially aminoglutethimide, Rogletimide, (.+-.)-Pyridoglutethimide, trilostane, testolactone, skin health king, vorozole, fadrozole, Anastrozole and letrozole.Exemestane can be used with for example its commercialization form such as trade mark AROMASIN.Formestane can be used with for example its commercialization form such as trade mark LENTARON.Fadrozole can be used with for example its commercialization form such as trade mark AFEMA.Anastrozole can be used with for example its commercialization form such as trade mark ARIMIDEX.Letrozole can be used with for example its commercialization form such as trade mark FEMARA or FEMAR.Aminoglutethimide can be used with for example its commercialization form such as trade mark ORIMETEN.The chemotherapeutant that comprises is that the present invention's combination of aromatase inhibitor is particularly useful for the hormone receptor positive tumor for the treatment of such as breast tumor.

Used term " antiestrogen " relates to the chemical compound at estrogen receptor level antagonism estrogen effect in the literary composition.This term including, but not limited to tamoxifen, fulvestrant, thunder cough up former times sweet smell and the hydrochloric acid thunder cough up the former times sweet smell.Tamoxifen can be used with for example its commercialization form such as trade mark NOLVADEX.The hydrochloric acid thunder is coughed up the former times sweet smell and can be used with for example its commercialization form such as trade mark EVISTA.Fulvestrant can be according to US 4,659, and 516 prescriptions of announcing make or it can be used with for example its commercialization form such as trade mark FASLODEX.The chemotherapeutant that comprises is that the present invention's combination of antiestrogen is particularly useful for the estrogen receptor positive tumors for the treatment of such as breast tumor.

Used term " androgen antagonist " relates to any material that can suppress the androgen biological effect in the literary composition, and including, but not limited to can be according to US 4,636, the bicalutamide (CASODEX) that 505 prescriptions of announcing are made.

Used term in the literary composition " GnRF agonist " is including, but not limited to 1: PN: WO02056903 PAGE: 25 claimed protein, goserelin and goserelin acetate.Goserelin is at US 4,100, obtains announcing in 274, and can use with for example its commercialization form such as trade mark ZOLADEX.1: PN: WO02056903 PAGE: 25 claimed protein can be according to US 5,843, and the prescription of announcing in 901 is made.

Used term in the literary composition " topoisomerase I inhibitor " is including, but not limited to hycamtin, gimatecan, Irinotecan, camptothecine and analog thereof, 9-nitrocamptothecin and macromolecular camptothecin conjugates PNU-166148 (compd A 1 among the WO 99/17804).Irinotecan can be used with for example its commercialization form such as trade mark CAMPTOSAR.Hycamtin can be used with for example its commercialization form such as trade mark HYCAMTIN.

Used term in the literary composition " topoisomerase II inhibitor " is including, but not limited to such as doxorubicin (comprising Liposomal formulation, as CAELYX), daunorubicin, epirubicin, idarubicin and Nemorubicin, mitoxantrone and losoxantrone (anthraquinone) and etoposide and teniposide (podophyllotoxin).Etoposide can be used with for example its commercialization form such as trade mark ETOPOPHOS.Teniposide can be used with for example its commercialization form such as trade mark VM 26-BRISTOL.Doxorubicin can be used with for example its commercialization form such as trade mark ADRIBLASTIN or ADRIAMYCIN.Epirubicin can be used with for example its commercialization form such as trade mark FARMORUBICIN.Idarubicin can be used with for example its commercialization form such as trade mark ZAVEDOS.Mitoxantrone can be used with for example its commercialization form such as trade mark NOVANTRON.

Term " microtubule active agent " relates to microtubule stabilizer, microtubule destabilizer and microtubule polymerization inhibitor, its including, but not limited to such as the taxane of paclitaxel and Duo Xi Ramulus et folium taxi cuspidatae, such as vinca alkaloids, the especially vincristine sulfate of vinblastine, especially vinblastine sulfate salt vincristine, vinorelbine, discodermolide, colchicine and such as the epothilone and the derivant thereof of epothilone B or D or derivatives thereof.Paclitaxel can be used with for example its commercialization form such as trade mark TAXOL.Many western Ramulus et folium taxi cuspidataes can be used with for example its commercialization form such as trade mark TAXOTERE.Vinblastine sulfate can be used with for example its commercialization form such as trade mark VINBLASTIN R.P.Vincristine sulfate can be used with for example its commercialization form such as trade mark FARMISTIN.Discodermolide can make according to the prescription that for example US 5,010,099 announces.Also comprise the Epothilone derivant of announcing among WO 98/10121, US 6,194,181, WO 98/25929, WO98/08849, WO99/43653, WO 98/22461 and the WO 00/31247.Preferred especially Epothilone A and/or B.

Used term " alkylating agent " is including, but not limited to cyclophosphamide, ifosfamide, melphalan or nitroso ureas (BCNU or Gliadel) in the literary composition.Cyclophosphamide can be used with for example its commercialization form such as trade mark CYCLOSTIN.Ifosfamide can be used with for example its commercialization form such as trade mark HOLOXAN.

Term " histone deacetylase inhibitors " or " hdac inhibitor " relate to the inhibition of histone deacetylase and have the chemical compound of antiproliferative activity.This comprises among the WO 02/22577 chemical compound, especially the N-hydroxyl-3-[4-[[(2-ethoxy of announcing) [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]-2E-2-acrylamide and officinal salt thereof.It also comprises Vorinostat (SAHA) especially.

Term " antineoplastic antimetabolite " is including, but not limited to 5-fluorouracil or 5-FU, capecitabine, gemcitabine, the agent of DNA demethylation, for example 5-azacitidine and decitabine, methotrexate and edatrexate and such as the antifol of pemetrexed.Capecitabine can be used with for example its commercialization form such as trade mark XELODA.Gemcitabine can be used with for example its commercialization form such as trade mark GEMZAR.Also comprise the monoclonal antibody trastuzumab, it can be used with for example its commercialization form such as trade mark HERCEPTIN

Used term " platinum compounds " is including, but not limited to carboplatin, cisplatin, cisplatin and oxaliplatin in the literary composition.Carboplatin can be used with for example its commercialization form such as trade mark CARBOPLAT.Oxaliplatin can be used with for example its commercialization form such as trade mark ELOXATIN.

Term used herein " chemical compound of targeting/reduction protein or lipid kinase activity or protein or lipid phosphatase activity or further the chemical compound of angiogenesis inhibitor " is including, but not limited to protein tyrosine kinase inhibitors and/or serine and/or threonine kinase enzyme inhibitor or lipid kinase inhibitors, for example:

A) targeting, reduction or suppress the platelet-derived active chemical compound of growth factor receptors (PDGFR), for example targeting, reduction or suppress the active chemical compound of PDGFR, especially the chemical compound that suppresses pdgf receptor, N-phenyl-2-pyrimidine-amine derivatives for example is as imatinib, SU101, SU6668 and GFB-111;

B) targeting, reduce or be suppressed to the active chemical compound of bfgf receptor (FGFR);

C) targeting, reduction or the active chemical compound of inhibition IGF-1 I (IGF-IR), for example targeting, reduction or suppress the active chemical compound of IGF-IR, especially the chemical compound that suppresses the IGF-IR receptor is as those chemical compounds of announcing among the WO 02/092599;

D) chemical compound of targeting, reduction or inhibition Trk receptor tyrosine kinase family active;

E) chemical compound of targeting, reduction or inhibition Axl receptor tyrosine kinase family active;

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

G) chemical compound of targeting, reduction or inhibition Kit/SCFR receptor tyrosine kinase activity;

H) targeting, reduction or the active chemical compound of inhibition C-kit receptor tyrosine kinase (part of PDGFR family), for example targeting, reduction or suppress the chemical compound of c-Kit receptor tyrosine kinase family active, especially the chemical compound that suppresses the c-Kit receptor, for example imatinib;

I) targeting, reduction or inhibition c-Abl family member and the active chemical compound of gene fusion product (for example BCR-Abl kinases) thereof, for example targeting, reduction or suppress the chemical compound of c-Abl family member and gene fusion product thereof, N-phenyl-2-pyrimidine-amine derivatives for example, for example imatinib, PD180970, AG957, NSC 680410 or from the PD173955 of ParkeDavis;

J) active and those US 5 especially of Raf family member, MEK, SRC, JAK, FAK, PDK and the Ras/MAPK family member of targeting, reduction or Profilin kinase c (PKC) and serine/threonine kinase or PI (3) kinases family or PI (3) kinases associated kinase family member and/or the member of cell cycle protein dependent kinase family (CDK), the chemical compound of announcing in 093,330 such as the staurosporine derivatives of midostaurin; Other examples for compounds also comprises for example UCN-01, Safingol, BAY 43-9006, bryostatin I, perifosine, llmofosine, RO 318220 and RO 320432; Such as those isoquinoline compounds, FTI, PD184352 or the QAN697 (P13K inhibitor) that announce among the WO 00/09495;

K) targeting, reduction or the active chemical compound of Profilin matter tyrosine kinase inhibitor, for example targeting, reduction or Profilin matter tyrosine kinase inhibitor are active, the chemical compound that comprises imatinib mesylate (GLEEVEC) or tyrphostin.The compound or pharmaceutically acceptable salt thereof of the preferred low-molecular-weight of tyrphostin (Mr＜1500), especially be selected from the chemical compound of benzal Cyanoacetyl-Cyacetazid class or S-aryl phenylpropyl alcohol dintrile or Double bottom thing quinolines (bisubstrate quinoline class), more particularly be selected from tyrphostin A23/RG-50810, AG 99, tyrphostin AG 213, tyrphostin AG 1748, tyrphostin AG 490, tyrphostin B44, tyrphostin B44 (+) enantiomer, tyrphostin AG 555, AG 494, tyrphostin AG 556, AG957 and adaphostin (4-{[(2,5-dihydroxy phenyl) methyl] amino }-benzoic acid diamantane (obsolete) ester, NSC 680410, adaphostin) chemical compound; And

I) targeting, reduce or suppress the epidermal growth factor family (EGFR of homodimer or heterodimer of receptor tyrosine kinase, ErbB2, ErbB3, ErbB4) active chemical compound, targeting for example, reduction or the active chemical compound of inhibition Epidermal Growth Factor Receptor Family especially suppress the receptor such as EGF, ErbB2, the EGF receptor tyrosine kinase family member of ErbB3 and ErbB4 or in conjunction with the chemical compound of EGF or EGF associated ligands, protein or antibody, and chemical compound general and that announce especially among those WO97/02266 particularly, protein or monoclonal antibody, the for example chemical compound of embodiment 39 or EP 0 564 409, WO 99/03854, EP 0520722, EP 0 566226, EP 0 787 722, EP 0 837 063, US 5,747,498, WO98/10767, WO97/30034, WO 97/49688, WO 97/38983, and WO 96/30347 (for example chemical compound of called after CP 358774) especially, chemical compound among WO 96/33980 (for example chemical compound ZD 1839) and the WO 95/03283 (for example chemical compound ZM105180), Si Tuman cloth (HERCEPTIN) for example, Cetuximab, Iressa, Tarceva, OSI-774, 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 7H-pyrrolo--[2,3-d] pyrimidine derivatives of announcing among the WO 03/013541.

Further the chemical compound of angiogenesis inhibitor comprises that their activity has another mechanism, for example suppresses irrelevant chemical compound, for example Thalidomide (THALOMID) and TNP-470 with protein or lipid kinase.

The chemical compound of targeting, reduction or Profilin matter or lipid phosphatase activity is the inhibitor such as phosphatase 1, phosphatase 2A, PTEN or CDC25, for example okadaic acid or derivatives thereof.

The chemical compound of inducing cell atomization is tretinoin, α-γ-or betatocopherol or α-γ-or δ-tocotrienol for example.

Used term cyclooxygenase-2 inhibitor replaces including, but not limited to for example Cox-2 inhibitor, 5-alkyl in the literary composition 2-arylamino phenylacetic acid and derivant, for example celecoxib (CELEBREX), rofecoxib (VIOXX), etoricoxib, valdecoxib or 5-alkyl-2-arylaminophenylacetiacids acids, for example 5-methyl-2-(2 '-chloro-6 '-fluoroanilino) phenylacetic acid, lumiracoxib.

Term used herein " diphosphate " is including, but not limited to etridonic acid, clodronicacid, for Shandong phosphoric acid, Pamidronic Acid, Alendronic Acid, her class's phosphoric acid, sharp plug phosphoric acid and Zoledronate." Etridonic acid " can use with for example its commercialization form such as trade mark DIDRONEL." Clodronic acid " can use with for example its commercialization form such as trade mark BONEFOS." for Shandong phosphoric acid " can use with for example its commercialization form such as trade mark SKELID." Pamidronic Acid " can with for example its such as trade mark AREDIA ^TMThe commercialization form use." Alendronic Acid " can be used with for example its commercialization form such as trade mark FOSAMAX." her class's phosphoric acid " can be used with for example its commercialization form such as trade mark BONDRANAT." sharp plug phosphoric acid " can be used with for example its commercialization form such as trade mark ACTONEL." Zoledronate " can be used with for example its commercialization form such as trade mark ZOMETA

Term " mTOR inhibitor " relates to mammal target spot (mTOR) that suppresses rapamycin and the chemical compound with antiproliferative activity, for example sirolimus (Rapamune ), everolimus (Certican ^TM), CCI-779 and ABT578.

Term used herein " heparanase inhibitors " is meant targeting, reduction or suppresses the chemical compound of heparin sulfate degraded.This term is including, but not limited to PI-88.

Term used herein " biological response modifier " is meant lymphokine or interferon, for example interferon gamma.

Term such as H-Ras, K-Ras or N-Ras used herein " the carcinogenic isotype inhibitor of Ras " is meant targeting, reduction or suppresses the chemical compound of Ras carcinogenic activity, for example " farnesyl transferase inhibitor ", for example L-744832, DK8G557 or P115777 (Zarnestra).

Term used herein " telomerase inhibitor " is meant targeting, reduction or suppresses the active chemical compound of telomerase.Targeting, reduction or the active chemical compound of inhibition telomerase especially suppress the chemical compound of telomerase receptor, for example telomestatin.

Term used herein " methionine aminopeptidase inhibitor " is meant targeting, reduction or suppresses the active chemical compound of methionine aminopeptidase.Targeting, reduction or the active chemical compound of inhibition methionine aminopeptidase be the bengamide or derivatives thereof for example.

Term used herein " proteasome inhibitor " is meant targeting, reduction or the active chemical compound of Profilin enzyme body.Targeting, reduction or the active chemical compound of Profilin enzyme body comprise for example PS-341 and MLN 341.

Term used herein " matrix metallo-proteinase inhibitor " or " MMP " inhibitor comprise but are not limited to collagen peptide simulation inhibitor and non-peptide simulation inhibitor, tetracycline derivant, for example hydroxamic acid peptide simulation inhibitor batimastat and oral biological effectiveness analog Marimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551), BMS-279251, BAY 12-9566, TAA211, MM1270B or AAJ996 thereof.

Term used herein " reagent that is used for the treatment of the blood malignant tumor " is including, but not limited to FMS sample tyrosine kinase inhibitor (for example targeting, reduction or suppress the active chemical compound of FMS sample tyrosine kinase receptor (Flt-3R)), interferon, 1-b-D-arabinofuranosyl adenin cytosine (ara-c) and bisulphate (bisulfan) and such as the ALK inhibitor of targeting, reduction or the kinase whose chemical compound of inhibition nerve lymphoma.

Targeting, reduction or the active chemical compound of inhibition FMS sample tyrosine kinase receptor (Flt-3R) especially suppress unit of Flt-3R receptor kinase family chemical compound, protein or antibody, for example PKC412, midostaurin, staurosporine derivatives, SU11248 and MLN518.

Term used herein " HSP90 inhibitor " is including, but not limited to the chemical compound of the endogenous atpase activity of targeting, reduction or inhibition HSP90; Chemical compound by the degraded of ubiquitin protein enzyme body approach, targeting, reduction or inhibition HSP90 service albumen (HSP90 client protein).The chemical compound of targeting, reduction or the endogenous atpase activity of inhibition HSP90 especially suppresses chemical compound, protein and the antibody of the atpase activity of HSP90, for example a kind of geldanamycin derivant 17-allyl amino, 17-de-methoxy geldanamycin (17AAG), other chemical compound, radicicol inhibitor and hdac inhibitor that geldanamycin is relevant.

Term used herein " anti proliferative antibody " is including, but not limited to trastuzumab (Herceptin ^TM), trastuzumab-DM1, erlotinib (Tarceva ^TM), bevacizumab (Avastin ^TM), Mabthera (Rituxan ), PR064553 (anti-CD40) and antibody 2C4.Multi-specificity antibody and antibody fragment that antibody is meant for example complete monoclonal antibody, polyclonal antibody, is made up of at least two kinds of complete antibodies are as long as they present the purpose biologic activity.

In order to treat acute myeloid leukemia (AML), can be with the aleukemic leukemia therapy associating of formula (I) chemical compound and standard, especially with the therapy associating that is used for the treatment of AML.Particularly, for example can unite the formula of taking (I) chemical compound and farnesyl transferase inhibitor and/or other such as daunorubicin, doxorubicin, Ara-C, VP-16, teniposide, mitoxantrone, idarubicin, carboplatin and PKC412 to the useful medicine of treatment AML.

Term " antileukemie chemical compound " comprises for example Ara-C, pyrimidine analogue, described pyrimidine analogue be 2 of deoxycytidine '-Alpha-hydroxy ribose (cytosine arabinoside) derivant.The purine analogue that also comprises hypoxanthine, Ismipur (6-MP) and fludarabine phosphate.

The chemical compound of targeting, reduction or inhibition of histone deacetylase (HDAC) inhibitor activity, for example sodium butyrate and Vorinostat (SAHA) suppress to be called the enzymatic activity of histidine deacetylase.Specific hdac inhibitor comprises MS275, SAHA, FK228 (FR901228 in the past), Trichostatin A (Trichostatin A) and US 6,552, the chemical compound of announcing in 065, especially, N-hydroxyl-3-[4-[[[2-(2-Methyl-1H-indole-3-yl)-ethyl]-amino] methyl] phenyl]-2E-2-acrylamide or its officinal salt, and N-hydroxyl-3-[4-[(2-ethoxy) 2-(1H-indol-3-yl) ethyl]-amino] methyl] phenyl]-2E-2-acrylamide or its officinal salt, especially lactate.

The chemical compound of targeting, reduction or inhibition serine/threonine mTOR kinase activity especially suppresses unit of mTOR kinases family chemical compound, protein or antibody, for example RAD, RAD001, CCI-779, ABT578, SAR543, rapamycin and derivant thereof, the AP23573 from Ariad, Yi Weimosi (CERTICAN) and sirolimus.

The antagonist of srif receptor used herein is meant targeting, treatment or suppresses the reagent of srif receptor, for example octreotide and SOM230.

Tumor cell damage mode refers to the mode such as ionizing radiation.The ionizing radiation that term " ionizing radiation " expression above and hereinafter described takes place with electromagnetic radiation (for example X-ray and gamma-radiation) or particle (for example alpha particle and beta particle).Ionizing radiation is used for (but being not limited to) radiation therapy, and is known in this area.See Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, people such as Devita edit, and the 4th edition, Vol.1, pp.248-275 (1993).

Term EDG bonding agent used herein refers to the immunosuppressant that a class is regulated the lymphocyte recirculation, for example FTY720.

(Yi Moweisi RAD) is the proliferation signal inhibitor of recent studies on to CERTICAN, and it stops T cell and vascular smooth muscle cell proliferation.

The term ribonucleotide reductase inhibitor refers to including, but not limited to the pyrimidine or the purine nucleoside analogs of fludarabine and/or cytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, carat Qu Bin, Ismipur (especially with ara-C logotype treatment ALL) and/or pentostatin.Ribonucleotide reductase inhibitor is hydroxyurea or 2-hydroxyl-1H-iso-indoles-1 especially, the 3-derovatives, for example people such as Nandy is at Acta Oncologica, Vol.33, No.8, PL-1, the PL-2, PL-3, PL-4, PL-5, PL-6, PL-7 or the PL-8 that mention among the pp.953-961 (1994).

Term used herein " S-ademetionine decarboxylase inhibitor " is including, but not limited to US 5,461, the chemical compound of announcing in 076.

Also comprise especially among the WO 98/35958 announce such as 1-(4-chloroanilino)-4-(4-pyridylmethyl)-2,3-benzodiazine or its officinal salt (for example succinate), WO 00/09495, WO00/27820, WO 00/59509, WO 98/11223, the chemical compound of announcing among WO00/27819 and the EP 0 769 947, the monoclonal antibody of protein or VEGF, comprise that those are people such as Prewett, Cancer Res, Vol.59, pp.5209-5218 (1999), people such as Yuan, Proc Natl Acad SciUSA, Vol.93, pp.14765-14770 (1996), people such as Zhu, Cancer Res, Vol.58, people such as pp.3209-3214 (1998) and Mordenti, Toxicol Pathol, Vol.27, No.1, pp.14-21 (1999), the chemical compound of describing among WO 00/37502 and the WO 94/10202, protein or monoclonal antibody, comprise people Cell such as O ' Reilly, Vol.79, the angiostatin of describing among the pp.315-328 (1994) (ANGIOSTATIN), people such as O ' Reilly, Cell, the endostatin of describing among the Vol.88, pp.277-285 (1997) (ENDOSTATIN), the ortho-aminobenzoic acid amide, ZD4190, ZD6474, SU5416, SU6668, the antibody of bevacizumab or VEGF antibody or anti-vegf receptor (for example rhuMAb and RHUFab), VEGF is fit (for example Macugon), the FLT-4 inhibitor, the FLT-3 inhibitor, VEGFR-2IgG1 antibody, Angiozyme (RPI 4610) and Avastan.

Photodynamic therapy used herein refers to that the chemicals that uses some to be known as photosensitizing agent is treated or the therapy of prophylaxis of cancer.The example of photodynamic therapy comprises the therapy of the reagent of use such as VISUDYNE and porfimer sodium.

Stable (Angiostatic) steroid of blood vessel used herein refers to and stops or suppress the reagent that blood vessel takes place, for example anecortave, triamcinolone, hydrocortisone, 11-α-Biao hydrocortisone, 11-deoxidation skin (steroid) alcohol, 17 α-hydroxyprogesterone, corticosterone, desoxycortone, testosterone, estrone and dexamethasone.

The implant that comprises corticosteroid refers to the reagent such as fluocinolone acetonide, dexamethasone.

Other chemotherapeutics is including, but not limited to the reagent of plant alkaloid, hormone reagent and antagonist, biological response modifier, preferred lymphokine or interferon, antisense oligonucleotide or oligonucleotide derivative or miscellaneous reagent or tool other or unknown role mechanism.

The active agent structures of identifying by Code Number, adopted name or trade name can obtain from actual release's standard outline " Merck index " or the data base such as " international monopoly (Patents International) (IMS world publication) ".

Above-mentioned can being prepared and using according to the method that this field (in the document of for example above quoting) described with the chemical compound that formula (I) chemical compound is united use.

Further, formula (I) chemical compound also can be used for such as using hormone, especially radiating known treatment methods coupling.

Especially, formula (I) chemical compound can be used as radiosensitizer, especially for the tumor treatment that X-ray therapy is shown as the hyposensitiveness perception.

" associating " expression is with the fixing joint of a dosage unit form or multi-componently be used for co-administered test kit or its any associating, can use or allow the gametophyte of associating to show in the interval of cooperative effect (for example cooperative effect) simultaneously separately at the gametophyte of described multicomponent kit Chinese style (I) chemical compound and associating and use separately.

Embodiment

The following example helps to describe the present invention and does not limit its scope:

Abbreviation

The DMSO dimethyl sulfoxide

ES-MS EFI mass spectrum

The EtOAc ethyl acetate

The HPLC high pressure liquid chromatography (HPLC)

The mL milliliter

The NMR nuclear magnetic resonance, NMR

The RT room temperature

^At _RETMinute be the HPLC retention time (method A) of unit

^Bt _RETMinute be the HPLC retention time (method B) of unit

^Ct _RETMinute be the HPLC retention time (method C) of unit

^Dt _RETMinute be the HPLC retention time (method D) of unit

The TFA trifluoroacetic acid

The THF oxolane

TMSCI chlorination trimethyl silyl

When not providing temperature, be reflected at room temperature and carry out.

The ratio of solvent, for example the ratio of the solvent in eluent or the solvent mixture with volume by volume ( ^v/ _v) provide.

Synthetic

By using silica gel (Merck; 40-63 μ m) carries out flash chromatography.Use to wrap in advance and carry out thin layer chromatography by the flat board of silica gel (Merck 60 F254).Finish the detection of composition by ultraviolet (254nm).Use Nucleosil 100-3 C ₁₈HD 125 * 4.0mm post (1mL/min; 20-100%NeCN/0.1%TFA in 7 minutes) (method A), SpectraSystem SP8800/UV2000 use Nucleosil 100-5 C ₁₈AB 250 * 4.6mm post (2mL/min; 2-100%MeCN/0.1%TFA in 10 minutes) (method B), use Chromalith Speed ROD RP18 50-4.6mm post (Merck) (2mL/min; 2-100%MeCN/0.1%TFA in 2 minutes) (method C) or C82.1-50mm 3 μ m post (water) (2mL/min; 5-95%MeCN/0.1%TFA in 2 minutes) (method D) carries out HPLC on Agilent HP 1100.On Varian Gemini 400 or Bruker DRX 500 spectrometers, carry out as internal standard with tetraethyl silane ¹H-NMR measures.Chemical shift be expressed as derive from tetraethyl silane with ppm magnetic field and express coupling constant (J) with hertz (Hz).Obtain the EFI mass spectrum with Fisons Instruments VG Platform II.With B ü chi 510 fusing point measurement device fusing points.The solvent and the chemical compound that can get with commerce synthesize.

The HPLC condition of analyzing:

System 1

The CH of 7 minutes linear gradient 20-100% ₃CN (0.1%TFA) and H ₂O (0.1%TFA) adds 2 minutes 100%CH ₃CN (0.1%TFA); Detect 30 ℃ of flow velocity 1mL/min at 215nm.Pillar: Nucleosil 100-3C18HD (125 * 4mm).

System 2

The CH of 7 minutes linear gradient 2-100% ₃CN (0.1%TFA) and H ₂O (0.1%TFA) adds 2 minutes 100%CH ₃CN (0.1%TFA); Detect 30 ℃ of flow velocity 1mL/min at 215nm.Pillar: Nucleosil 100-3C18HD (125 * 4mm).

Embodiment 1

3-{7-amino-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidine-6-yl }-phenol

Be dissolved in 6-(3-benzyloxy-phenyl)-3-[4-(4-methyl-piperazine-1-yl)-phenyl of THF (6mL)]-pyrazolo [1,5-a] pyrimidin-7-yl amine (stage 1.1) (25mg, 0.051mmol) (10%Engelhard 4505,6mg) hydrogenation 13 hours under the condition there being Pd/C.Filter under the reduced pressure and evaporating solvent flash chromatography residue (silica gel, CH afterwards ₂Cl ₂/ MeOH/NH ₃=95: 5: 0.1) to obtain white solid chemical compound (14mg, the 0.035mmol of embodiment 1; 70%): ES-MS:M+H=401.1, R _f(CH ₂Cl ₂/ MeOH/NH ₃=90: 10: 0.1)=0.33, HPLC: ^At _Ret=2.77 minutes.

¹H-NMR (400MHz, DMSO-d ₆): 9.59 (s, 1H, OH), 8.58/8.18 (s/s, 1H/1H, pyrazolopyrimidine base), 8.01 (d, 9.0Hz, 2H, phenyl), 7.48 (s, 2H, NH ₂), 7.32 (t, 8.5Hz, 1H, phenyl-OH), 6.99 (d, 9.0Hz, 2H, phenyl), 6.96 (d, 8.5Hz, 1H, phenyl-OH), 6.93 (s, 1H, phenyl-OH), 6.80 (d, 8.5Hz, 1H, phenyl-OH), 3.17/2.48 (m/m, 4H/4H, piperazinyl), 2.24 (s, 3H, CH ₃).

Stages 1.1 6-(3-benzyloxy-phenyl)-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl amine

Stirring at room is dissolved in 4-(4-(4-methyl-piperazine-1-yl)-phenyl)-2H-pyrazole-3-yl amine (stage 1.2) (100mg of ethanol (1ml), 0.388mmol), 2-(3-benzyloxy-phenyl)-3-oxo-propionitrile (stage 1.3) (98mg, 0.388mmol), HCl (is dissolved in ethanol, 2.5mM; 1,55mmol, 0.9mL) 17 hours.Add H ₂O (4mL) and K ₂CO ₃Use CH (250mg) ₂Cl ₂(20mL, 2x) extractive reaction mixture.Use H ₂The organic facies that O (10mL) washing merges, dry (Na ₂SO ₄), under reduced pressure, concentrate and flash chromatography (silica gel, 2.5 * 15cm, CH ₂Cl ₂/ MeOH=9: 1) to obtain white solid chemical compound (60mg, the 0.122mmol in stage 1.1; 32%); ES-MS:M+H=491.0, R _f(CH ₂Cl ₂/ MeOH/NH ₃=90: 10: 0.1)=0.42; HPLC: ^At _Ret=4.69 minutes.

¹H-NMR (400MHz, DMSO-d ₆): 8.79/8.21 (s/s, 1H/1H, pyrazolopyrimidine base), 8.03 (d, 9.0Hz, 2H, phenyl), 7.53 (s, 2H, NH ₂), 7.44 (m, 5H, benzyls), 7.32 (t, 8.5Hz, 1H, phenyl-OH), 7.29 (s, 1H, phenyl-OH), 7.13 (d, 8.5Hz, 1H, phenyl-OH), 7.06 (d, 8.5Hz, 1H, phenyl-OH), 6.97 (d, 9.0Hz, 2H, phenyl), 5.19 (s, 2H, benzyls), 3.17/2.48 (m/m, 4H/4H, piperazinyl), 2.24 (s, 3H, CH ₃).

Stages 1.2 4-(4-(4-methyl-piperazine-1-yl)-phenyl)-2H-pyrazole-3-yl amine

Be dissolved in 2-[4-(4-methyl-piperazine-1-yl)-phenyl of AcOH 98 ℃ of stirrings]-3-oxo-propionitrile (stage 1.4) (370mg, 1.52mmol), a hydrazine hydrate (0.185mL, 3.8mmol) 3 hours.Add H behind the cool to room temperature ₂O (8mL) and dense HCl (0.8mL) and stirred reaction mixture 20 minutes under refluxad.Pass through slowly to add NH behind the cool to room temperature ₃(25%) adjusts compound of reaction to alkaline pH.Filter out deposit and preserve and be used for further purification.Use CH ₂Cl ₂(50mL, 3x) extractive reaction solution, dry (Na ₂SO ₄) and under reduced pressure, concentrate.Merge material and flash chromatography (silica gel, 3.0 * 18cm, CH sedimentary and extraction ₂Cl ₂/ MeOH/NH ₃=9: 1: 01) to obtain white solid chemical compound (277mg, the 1.08mmol in stage 1.2; 71%); ES-MS:M+H=258.1, R _f(CH ₂Cl ₂/ MeOH/NH ₃=90: 10: 0.1)=0.28; HPLC: ^At _Ret=4.33 minutes.

¹H-NMR (400MHz, DMSO-d ₆): 11.55 (s/broad, 1H, NH), 7.55 (s, 1H, pyrolyl), 7.35 (d, 9.0Hz, 2H, phenyl), 6.91 (d, 9.0Hz, 2H, phenyl), 4.55 (s/broad, 2H, NH ₂), 3.10/2.46 (m/m, 4H/4H, piperazinyl), 2.23 (s, 3H, CH ₃).

Stages 1.3 2-(3-benzyloxy-phenyl)-3-oxo-propionitrile

(260mg 11.3mmol) is dissolved in (11mL) in the dehydrated alcohol with Na in 20 minutes under argon environment.Add (3-benzyloxy-phenyl)-acetonitrile (1.9g, 8.68mmol) and Ethyl formate (1.05mL, 13.0mmol) after stirred reaction mixture 2 hours under refluxad.The evaporation under reduced pressure solvent adds H ₂O (20mL) also adjusts to pH=4.0 by adding AcOH and uses CH afterwards ₂Cl ₂(30mL, 2x) extractive reaction suspension.Use H ₂The organic facies that O (10mL) washing merges, dry (Na ₂SO ₄), under reduced pressure, concentrate and flash chromatography (silica gel, 4.5 * 25cm, CH ₂Cl ₂/ MeOH=98: 2) to obtain white solid chemical compound (780mg, the 3.11mmol in stage 1.3; 36%); ES-MS:M-H=250.0, R _f(CH ₂Cl ₂/ MeOH=95: 5)=0.49; HPLC: ^At _Ret=6.07 minutes.

¹H-NMR (400MHz, DMSO-d ₆): 7.45-7.25/6.98-6.88 (m/m, 8H, aryl), 5.09 (s, 2H, CH ₂), 3.98 (s, 2H, CH ₂).

Stages 1.4 2-[4-(4-methyl-piperazine-1-yl)-phenyl]-3-oxo-propionitrile

Under argon environment, in 10 minutes, 160mg Na (7.0mmol) is dissolved in (6mL) in the dehydrated alcohol.Add [4-(4-methyl-piperazine-1-yl)-phenyl]-acetonitrile (stage 1.5) (1g, 4.64mmol) and Ethyl formate (0.56mL, 7.0mmol) back stirred reaction mixture 1 hour under backflow.(50mL 3x) is dissolved in H with solid residue after the washing reaction slurry with ether ₂Adjust to pH=3.9 among the O (60mL) and by adding AcOH.Use CH ₂Cl ₂(50mL, 3x) extraction water solution.Use H ₂The organic facies that O (50mL) washing merges.Mix two waters and lyophilizing.From MeOH/CH ₂Cl ₂The residue that middle crystallization obtains provides white crystal chemical compound (721mg, the 3.0mmol in stage 1.4; 64%); ES-MS:M+H=244.1; HPLC: ^At _Ret=2.43 minutes.

¹H-NMR (400MHz, DMSO-d ₆): chemical compound in solution, form tautomeric equilibrium: 7.87/7.77 (s/s, 1H, CH=/CH-OH), 7.53/7.17 (d/d, 9.0Hz, 2H, phenyl), 7.84/7.82 (d/d, 9.0Hz, 2H, phenyl), (3.10 m, 4H, piperazinyl), (2.57/2.51 m/m, 4H, piperazinyl), 2.29/2.26 (s, 3H, CH ₃).

Stage 1.5 [4-(4-methyl-piperazine-1-yl)-phenyl]-acetonitrile

Under argon environment, be dissolved in 1 in 85 ℃ of stirrings, (4-bromo-the phenyl)-acetonitrile of 2-dimethoxy-ethane (70mL) (5g, 25.5mmol), 1-methyl-piperazine (3.4mL, 30.6mmol), K ₂CO ₃(7.68g, 35.7mmol), Pd (AcO) ₂(280mg, 1.275mmol), 2-(two-tert-butyl group phosphino-)-diphenyl (1.14g, 3.825mmol) 20 hours.Add H ₂O (100mL) uses CH afterwards ₂Cl ₂(100mL, 3x) extractive reaction mixture.Wash the organic facies of merging with water, dry (Na ₂SO ₄), under reduced pressure, concentrate and flash chromatography (silica gel, 4.5 * 34cm, CH ₂Cl ₂/ MeOH=95: 5) to obtain white solid chemical compound (2.8g, the 13mmol in stage 1.5; 51%); ES-MS:M+H=216.1; R _f(CH ₂Cl ₂/ MeOH=9: 1)=0.47; HPLC: ^At _Ret=2.24 minutes.

¹H-NMR (400MHz, DMSO-d ₆): 7.14/6.91 (d/d, 9.5Hz, 2H/2H, phenyl), 7.53 (s, 2H, NH ₂), 7.44 (m, 5H, benzyls), 7.32 (t, 8.5Hz, 1H, phenyl-OH), 7.29 (s, 1H, phenyl-OH), 3.84 (s, 2H, benzyls), 3.09/2.42 (t/t, 5.0Hz, 4H/4H, piperazinyl), 2.18 (s, 3H, CH ₃).

Embodiment 2

6-(3-methoxyl group-phenyl)-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl amine

Chemical compound and the synthetic 6-(3-methoxyl group-phenyl) of 2-(3-methoxyl group-phenyl)-3-oxo-propionitrile (stage 2.1)-3-[4-(4-methyl-piperazine-1-yl)-phenyl by the condensation stage 1.2 similarly with the preparation of the chemical compound of embodiment 1]-pyrazolo [1,5-a] pyrimidin-7-yl amine.Productive rate: 48%, pressed powder; ES-MS:M+H=415.1; HPLC: ^At _Ret=3.45 minutes.

¹H-NMR (400MHz, DMSO-d ₆): 8.59/8.23 (s/s, 1H/1H, pyrazolopyrimidine base), 8.06 (d, 9.0Hz, 2H, phenyl), 7.55 (s, 2H, NH ₂), 7.43 (t, 8.5Hz, 1H, phenyl-OMe), 7.10 (d, 8.5Hz, 1H, phenyl-OMe), 7.08 (s, 1H, phenyl-OMe), 6.80 (d, 8.5Hz, 1H, phenyl-OMe), 6.98 (d, 9.0Hz, 2H, phenyl), 3.83 (s, 3H, CH ₃-O), 3.16/2.47 (m/m, 4H/4H, piperazinyl), 2.25 (s, 3H, CH ₃).

Stages 2.1 2-(3-methoxyl group-phenyl)-3-oxo-propionitrile

With the preparation of the chemical compound in stage 1.3 Synthetic 2-(3-methoxyl group-phenyl)-3-oxo-propionitrile similarly: productive rate: 76%; White powder; ES-MS:M-H=174.0; HPLC: ^At _Ret=4.75 minutes.

¹H-NMR (400MHz, DMSO-d ₆): chemical compound in solution, form tautomeric equilibrium: 8.09/7.67 (s/s, 1H, CH=/CH-OH), 7.38-7.23 (m, 2H, phenyl), 7.01-6.97 (m, 1H, phenyl), 6.88-6.79 (m, 1H, phenyl), 3.74 (s/broad, 3H, CH ₃-O).

Embodiment 3

6-(3,5-dimethoxy-phenyl)-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidine-7-

Base amine

Chemical compound and the 2-(3 by the condensation stage 1.2 similarly with the preparation of the chemical compound of embodiment 1,5-dimethoxy-phenyl)-the synthetic 6-(3 of 3-oxo-propionitrile (stage 3.1), 5-dimethoxy-phenyl)-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl amine.Productive rate: 44%, pressed powder; ES-MS:M+H=445.0; HPLC: ^At _Ret=3.77 minutes.

¹H-NMR (400MHz, DMSO-d ₆): 8.59/8.23 (s/s, 1H/1H, pyrazolopyrimidine base), 8.06 (d, 9.0Hz, 2H, phenyl), 7.55 (s, 2H, NH ₂), 7.43 (t, 8.5Hz, 1H, phenyl-OMe), 7.10 (d, 8.4Hz, 1H, phenyl-OMe), 7.57 (s, 2H, NH ₂), 7.01 (d, 9.0Hz, 2H, phenyl), 6.89 (s, 2H, phenyl-OMe), 6.54 (s, 1H, phenyl-OMe), 3.83 (s, 6H, CH ₃-O), 3.16/2.47 (m/m, 4H/4H, piperazinyl), 2.24 (s, 3H, N-CH ₃).

Stages 3.1 2-(3,5-dimethoxy-phenyl)-3-oxo-propionitrile

With Synthetic 2-(3,5-dimethoxy-phenyl)-3-oxo-propionitrile like the compounds in stage 1.3.Productive rate: 48%; White powder; ES-MS:M+H=206.0; HPLC: ^At _Ret=4.79 minutes.

¹H-NMR (400MHz, DMSO-d ₆): chemical compound in solution, form tautomeric equilibrium: 8.11/7.68 (s/s, 1H, CH=/CH-OH), 6.85/6.54 (s/s, 2H, phenyl), 6.44/6.38 (s/s, 1H, phenyl), 3.74 (s/broad, 6H, CH ₃-O).

Embodiment 4

6-(3-benzyloxy-phenyl)-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl amine is by disclosed step preparation of stages 1.1.

Embodiment 5-69

With the synthetic similarly the following example of listing in table 1 of the preparation of embodiment 1.Described the synthetic of the intermediate that is used to prepare embodiment 5-69 chemical compound in the text below the table 1, wherein said embodiment 5-69 chemical compound is commercial non-availability.Carry in the situation of free amine group (embodiment 52-54) at title compound, end product produced from the precursor that it has nitro accordingly by the hydrogenization under the Pd/C in having THF/MeOH (10%) condition in a few hours.

Table 1.

Numbering

A

R2

R3

Analytical data

Numbering	A	R2	R3	Analytical data
					5	4-(4-methyl-piperazine-1-yl) phenyl	The 4-chlorphenyl	H	ES-MS [M+1] +＝ 419.0/421. 0； HPLC At Ret=3.71 minutes
6	4-(4-methyl-piperazine-1-yl) phenyl	The 3-chlorphenyl	H	ES-MS [M+1] +＝ 419.0/421. 0； HPLC At Ret=3.92 minutes
					7	4-(4-methyl-piperazine-1-yl) phenyl	Phenyl	H	ES-MS [M+1] +＝ 385.1； PLC At Ret=3.31 minutes
8	4-(4-methyl-piperazine-1-yl) phenyl	Phenyl	Methyl	ES-MS [M+1] +＝ 399.1； HPLC At Ret=3.34 minutes

Numbering	A	R2	R3	Analytical data
					12	Phenyl	The 4-methoxyphenyl	Methyl	ES-MS [M+1] +＝ 331.1； HPLC Bt Ret=6.4 minutes
13	4-methoxyl group-phenyl	Phenyl	Methyl	ES-MS [M+1] +＝ 331.1； HPLC Bt Ret=6.3 minutes
					14	4-methoxyl group-phenyl	The 4-bromophenyl	Methyl	ES-MS [M+1] +＝ 408.9/410. 9； HPLC Bt Ret=7.1 minutes
15	Phenyl	The 4-bromophenyl	Methyl	ES-MS [M+1] +＝ 378.9/380. 9； HPLC Bt Ret=7.0 minutes
					16	Phenyl	2, the 6-Dichlorobenzene base	H	ES-MS [M+1] +＝ 354.9/356. 9； HPLC Bt Ret=7.9 minutes

Numbering	A	R2	R3	Analytical data
					27	4-(4-methyl-piperazine-1-yl) phenyl	The 2-methoxyphenyl	H	ES-MS [M+1] +＝ 415.2； HPLC Ct Ret=1.75 minutes
28	Pyridin-3-yl	The 3-methoxyphenyl	H	ES-MS [M+1] +＝ 318.6； HPLC Ct Ret=1.84 minutes
					29	3-(4-methyl-piperazine-1-yl) phenyl	The 3-hydroxy phenyl	H	ES-MS [M+1] +＝ 401.6； HPLC At Ret=1.78 minutes
30	2-methoxyl group-5-(4-methyl-piperazine-1-yl) phenyl	3-benzyloxy phenyl	H	ES-MS [M+1] +＝ 521.3； HPLC Ct Ret=2.07 minutes
					31	2-methoxyl group-5-(4-methyl-piperazine-1-yl) phenyl	The 3-hydroxy phenyl	H	ES-MS [M+1] +＝ 431.7； HPLC Ct Ret=1.66 minutes

Numbering	A	R2	R3	Analytical data
					32	4-(4-methyl-piperazine-1-yl) phenyl	2-benzyloxy phenyl	H	ES-MS [M+1] +＝ 491.2； HPLC Ct Ret=1.77 minutes
33	4-(4-methyl-piperazine-1-yl) phenyl	The 2-hydroxy phenyl	H	ES-MS [M+1] +＝ 401.2； HPLC Dt Ret=1.37 minutes
					34	4-(4-methyl-piperazine-1-yl) phenyl	4-benzyloxy phenyl	H	ES-MS [M+1] +＝ 491.2； HPLC Ct Ret=1.85 minutes
35	4-(4-methyl-piperazine-1-yl) phenyl	The 4-hydroxy phenyl	H	ES-MS [M+1] +＝ 401.2； HPLC Dt Ret=1.32 minutes
					36	3-(4-methyl-piperazine-1-yl) phenyl	2-benzyloxy phenyl	H	ES-MS [M+1] +＝ 491.3； HPLC At Ret=2.02 minutes

Numbering	A	R2	R3	Analytical data
					37	3-(4-methyl-piperazine-1-yl) phenyl	The 2-hydroxy phenyl	H	ES-MS [M+1] +＝ 401.3； HPLC At Ret=1.71 minutes
38	3-(4-methyl-piperazine-1-yl) phenyl	4-benzyloxy phenyl	H	ES-MS [M+1] +＝ 491.3； HPLC Ct Ret=2.05 minutes
					39	3-(4-methyl-piperazine-1-yl) phenyl	The 4-hydroxy phenyl	H	ES-MS [M+1] +＝ 401.3； HPLC Ct Ret=1.70 minutes
40	2-methoxyl group-5-(4-methyl-piperazine-1-yl) phenyl	2-benzyloxy phenyl	H	ES-MS [M+1] +＝ 521.3； HPLC Ct Ret=1.99 minutes
					41	2-methoxyl group-5-(4-methyl-piperazine-1-yl) phenyl	The 2-hydroxy phenyl	H	ES-MS [M+1] +＝ 431.3； HPLC Ct Ret=1.70 minutes

Numbering	A	R2	R3	Analytical data
					42	2-methoxyl group-5-(4-methyl-piperazine-1-yl) phenyl	4-benzyloxy phenyl	H	ES-MS [M+1] +＝ 521.3； HPLC Ct Ret=2.05 minutes
43	2-methoxyl group-5-(4-methyl-piperazine-1-yl) phenyl	The 4-hydroxy phenyl	H	ES-MS [M+1] +＝ 431.3； HPLC Ct Ret=1.68 minutes
					44	1-Methyl-1H-indole-3-base	3-benzyloxy phenyl	H	ES-MS [M+1] +＝ 446.2； HPLC Ct Ret=2.39 minutes
45	1-Methyl-1H-indole-3-base	The 3-hydroxy phenyl	H	ES-MS [M+1] +＝ 356.6； HPLC Ct Ret=2.06 minutes
					46	The 3-pyridine radicals	The 3-hydroxy phenyl	H	ES-MS [M+1] +＝ 304.1； HPLC Ct Ret=1.72 minutes

Stage 5.1:2-(4-chloro-phenyl)-3-oxo-propionitrile

Prepare 2-(4-chloro-phenyl)-3-oxo-propionitrile similarly with the preparation of the chemical compound in stage 1.3: 89%; ES-MS[M-1] ^-=177.9/179.9; HPLC ^At _Ret=5.67 minutes.

Stage 6.1:2-(3-chloro-phenyl)-3-oxo-propionitrile

Prepare 2-(3-chloro-phenyl)-3-oxo-propionitrile similarly with the preparation of the chemical compound in stage 1.3: 89%; ES-MS[M-1] ^-=177.9/179.9; HPLC ^At _Ret=5.60 minutes.

Stages 8.1 3-oxo-2-phenyl-butyronitrile

Prepare 3-oxo-2-phenyl-butyronitrile similarly with the preparation of the chemical compound in stage 1.3: 62%, white crystal, m.p.＞215 ℃; ES-ES-MS M-H=157.9, R _f(hexane/AcOEt=1: 1)=0.57.

¹H-NMR(400MHz，DMSO-d ₆)：7.84(d，9.0Hz，2H)，7.04(t，9.0Hz，2H)，6.68(t，9.0Hz，1H)，3.21(s/broad，1H，CH)，2.03(s，3H，CH ₃)。

Stages 9.1 2-methyl-3-oxo-3-phenyl-propionitrile

With people such as Yoo, Tetrahedron Lett., the 43rd volume, No.27, the method for 4813-4815 page or leaf (2002) prepares 2-methyl-3-oxo-3-phenyl-propionitrile similarly.Under argon environment in THF (15mL) stir 2-bromo-propionitrile (0.965mL, 11.05mmol) and indium powder (In-powder) (975mg, 8.5mmol) 1 hour.Add in 2 minutes the benzoyl nitrile (735mg, 5.6mmol) after, (Emrys optimizer, personal chemistry is Sweden) in 60 ℃ of stirred reaction mixtures 30 minutes in microwave oven.By Hyflo filter and with after THF (5mL) washing under reduced pressure concentrated reaction solution and at ether (150mL) and phosphate buffer (pH=7 separates between 150mL).Use ether (150mL) aqueous phase extracted after separating organic facies.With the organic facies that saline (30mL) washing merges, dry (Na ₂SO ₄), under reduced pressure, concentrate and flash chromatography (silica gel, 2 * 18cm, hexane/AcOEt=3: 1) obtain faint yellow oily chemical compound (300mg, the 1.9mmol in stage 9.1; 34%); ES-MS:M-H=157.9; R _f(hexane/AcOEt=1: 1)=0.60.

¹H-NMR(400MHz，DMSO-d ₆)：8.06(d，8.5Hz，2H)，7.74(t，8.5Hz，1H)，7.62(t，8.5Hz，2H)，5.17(q，8.5Hz，1H，CH)，1.52(s，3H，CH ₃)。

With the preparation of the chemical compound in stage 1.1 similarly by condensation 2,3-two chloro-N-[4-(cyano group-formyl-methyl)-phenyl]-chemical compound of benzsulfamide (stage 10.1) and the synthetic embodiment 10 of 4-(4-dimethylamino-phenyl)-2H-pyrazole-3-yl amine (stage 10.3).

Stage 10.1 2,3-two chloro-N-[4-cyano group-formyl-methyl]-phenyl]-benzsulfamide

At N ₂In 15 minutes, add the new fritter sodium that downcuts under the environment in batches and (be total to 2.3g, 100mmol) in anhydrous EtOH (230mL), its meeting slight exotherm (reaching 43 ℃).All sodium dissolving back (about 1 hour) adds 2 in room temperature in colourless solution, 3-two chloro-N-(4-cyano methyl-phenyl)-benzsulfamide (stage 10.2) (26.27g, 77mmol) and Ethyl formate (11.2mL, 139mmol).Mixture heated was refluxed 2 hours.Under reduced pressure, remove behind the cool to room temperature and desolvate and residue is dissolved in H ₂Among the O (20mL), add AcOH (200mL then; PH 4).Use CH ₂Cl ₂(2x, 500mL) aqueous layer extracted is used H ₂The Organic substance that the O washing merges also passes through Na ₂SO ₄Dry.Finish purification (silica gel, EtOAc and CH by chromatography repeatedly ₂Cl ₂/ MeOH=98: 2) crystalline 2 to obtain ecru, 3-two chloro-N-[4-(cyano group-formyl-methyl)-phenyl]-benzsulfamide (233mg, productive rate 1%): m.p.88-102 ℃; (CH ₂Cl ₂/ MeOH=95: 5): 0.22; ES-MS[M+1] ⁺=368; HPLC ^Bt _Ret=5.61 minutes.

Stage 10.2 2,3-, two chloro-N-(4-cyano methyl-phenyl)-benzsulfamide

In 20 minutes, add room temperature and be dissolved in 4-aminobenzyl cyanide (12g, 90.8mmol) solution and be dissolved in 2 of THF (80mL), 3-dichloro-benzenes-sulfonyl chloride (22.29g, 90.8mmol) solution of pyridine (11mL).Backflow stirring reaction 2 hours.Cooling back is removed under reduced pressure and is desolvated and remaining solid is suspended from (200mL) among the 10%HCl.Filter out the crystal crude product, use H ₂O washing and 60 ℃ of dryings.By crude compound being suspended from (250mL) among the MeOH, reflux is filtered and drying is finished final purification.Obtain 2 with orange crystal, 3-two chloro-N-(4-cyano methyl-phenyl)-benzsulfamide (26.54g, 86%): m.p:202-206 ℃; (CH ₂Cl ₂/ MeOH 98: 2): 0.54; ES-MS[M-1] ^-=338.8; HPLC ^Bt _Ret=5.85 minutes.

Stages 10.3 4-(4-dimethylamino-phenyl)-2H-pyrazole-3-yl amine

As U.S. Patent number 2,989,539 (20.6.61; Anderson and Reiff; Embodiment 18) described in prepare 4-(4-dimethylamino-phenyl)-2H-pyrazole-3-yl amine from 2-(4-dimethylamino-phenyl)-3-oxo-propionitrile (stage 10.4) and hydrazine hydrate.4-(4-dimethylamino-phenyl)-2H-pyrazole-3-yl amine: m.p.173-176 ℃; (CH ₂Cl ₂/ MeOH/NH ₃=90: 10: 1): 0.37; ES-MS[M+1] ⁺=203; HPLC ^Bt _Ret=1.40 minutes.

Stages 10.4 2-(4-dimethylamino-phenyl)-3-oxo-propionitrile

As U.S. Patent number 2,989, prepare 2-(4-dimethylamino-phenyl)-3-oxo-propionitrile from (4-dimethylamino-phenyl)-acetonitrile, Ethyl formate and sodium described in 539 (embodiment 18).

2-(4-dimethylamino-phenyl)-3-oxo-propionitrile: m.p.175-178 ℃; ES-MS[M+1] ⁺=189; HPLC ^Bt _Ret=2.00 minutes.

The synthetic chemical compound of using 4-(4-dimethylamino-phenyl)-2H-pyrazole-3-yl amine (stage 10.3) and 4-chloro-benzenesulfonic acid 4-(cyano group-formyl-methyl)-phenylester (stage 11.1) to prepare embodiment 11 similarly with the chemical compound of embodiment 10.

Stages 11.1 4-chloro-benzenesulfonic acid 4-(cyano group-formyl-methyl)-phenylester

As described in embodiment 10 (stage 10.1), prepare 4-chloro-benzenesulfonic acid 4-(cyano group-formyl-methyl)-phenylester, but use is 4-(cyano methyl) phenyl-4-chlorobenzene-1-sulfonate that commerce can get.

4-chloro-benzenesulfonic acid 4-(cyano group-formyl-methyl)-phenylester (162mg); Faint yellow solid; (CH ₂Cl ₂/ MeOH=95: 2): 0.32; ES-MS[M+1] ⁺=335; HPLC ^Bt _Ret=6.23 minutes.

Stages 12.1 2-(4-methoxyl group-phenyl)-3-oxo-butyronitrile

As Smith, Breen, Hajek and Awang, J.Org.Chem., the 35th volume, No.7, the described preparation of 2215-2221 page or leaf (1970) 2-(4-methoxyl group-phenyl)-3-oxo-butyronitrile.

Stages 14.1 2-(4-bromo-phenyl)-3-oxo-butyronitrile

According to Rau, Ger.Offen., the method Synthetic 2 of DE 3001266 (1980)-(4-bromo-phenyl)-3-oxo-butyronitrile.

Stages 16.1 1,2-(2,6-two chloro-phenyl)-3-oxo-propionitrile

As Menzer, Lankau and Unverferth, Ger.Offen., DE 19521822 (1996) described preparations 1,2-(2,6-two chloro-phenyl)-3-oxo-propionitrile.

Stages 17.1 4-(3-methoxyl group-phenyl)-2H-pyrazole-3-yl amine

As people such as Bruni, Heterocyclic.Chem., the 32nd volume, No.1, the described preparation of 291-298 page or leaf (1995) 4-(3-methoxyl group-phenyl)-2H-pyrazole-3-yl amine and stage 22.2.

Stages 19.1 2-benzo [b] thiene-3-yl--3-oxo-propionitrile

With the preparation of the chemical compound in stage 1.3 Synthetic 2-benzo [b] thiene-3-yl--3-oxo-propionitrile similarly: productive rate: 56%; White powder; ES-MS:M-H=123.9; HPLC: ^At _Ret=2.20 minutes.

¹H-NMR(300MHz，DMSO-d ₆)：12.0(s/broad，1H)，8.00-7.70(m，3H)，7.45-7.35(m，2H)。

Stages 21.1 3-oxo-2-thiene-3-yl--propionitrile

With the synthetic similarly 3-oxo of the preparation of the chemical compound in stage 1.3-2-thiene-3-yl--propionitrile: productive rate: 51%; White powder, ES-MS:M-H=112.9; HPLC: ^At _Ret=2.03 minutes.

Chemical compound forms tautomeric equilibrium in solution: ¹H-NMR (300MHz, DMSO-d ₆): 7.95/7.55 (s/s, 1H, CH=/CH-OH), 7.55-7.50 (m, 2H), 7.30-7.20 (m, 1H).

Stages 22.1 4-benzo [b] thiene-3-yl--1H-pyrazole-3-yl amine

With synthetic similarly 4-benzo [b] thiene-3-yl-of the preparation of the chemical compound in stage 1.2-1H-pyrazole-3-yl amine: productive rate: 80%; White powder; ES-MS:M+H=216.0.

¹H-NMR(300MHz，DMSO-d ₆)：12.0(s/broad，1H)，8.00-7.80(m，2H)，7.75(s/broad，1H)，7.60(s/broad，1H)，7.40-7.30(m，2H)。

Stage 22.2 (2-(3-methoxyl group-phenyl)-3-oxo-propionitrile)

As people such as Bruni, Heterocyclic.Chem., the 32nd volume, No.1, the described preparation of 291-298 page or leaf (1995) (2-(3-methoxyl group-phenyl)-3-oxo-propionitrile).

Stages 23.1 2-formyl-3-phenyl-propionitrile

With the preparation of the chemical compound in stage 1.3 Synthetic 2-formyl-3-phenyl-propionitrile similarly: productive rate: 77%; Oily; ES-MS:M-H=158.0.

Chemical compound forms tautomeric equilibrium in solution: ¹H-NMR (300MHz, DMSO-d ₆): 7.40-7.15 (m, 5H), 2.85-2.75 (m, 2H).

Stages 24.1 4[3-(4-methyl-piperazine-1-yl)-phenyl]-acetonitrile

With the synthetic similarly 4[3-(4-methyl-piperazine-1-yl) of the preparation of the chemical compound in stage 1.5-phenyl]-acetonitrile: productive rate: 55%; Brown solid; ES-MS:M+H=216.7; HPLC: ^Ct _Ret=1.65 minutes.

¹H-NMR(300MHz，CDCl ₃)：7.30-7.25(m，1H)，6.90-6.82(m，2H)，6.80-6.75(m，1H)，3.70(s，2H)，3.25-3.15(m，4H)，2.60-2.50(m，4H)，2.35(s，3H)。

Stages 24.2 2-[3-(4-methyl-piperazine-1-yl)-phenyl]-3-oxo-propionitrile

With the preparation of the chemical compound in stage 1.3 Synthetic 2-[3-(4-methyl-piperazine-1-yl)-phenyl]-3-oxo-propionitrile similarly: productive rate: 100%; Brown solid; ES-MS:M+H=244.1; HPLC: ^Ct _Ret=1.67 minutes.

Stages 24.3 4-[3-(4-methyl-piperazine-1-yl)-phenyl]-1H-pyrazole-3-yl amine

With the synthetic similarly 4-[3-(4-methyl-piperazine-1-yl) of the preparation of the chemical compound in stage 1.2-phenyl]-1H-pyrazole-3-yl amine: productive rate: 36%; Yellow foam; ES-MS:M+H=258.2; HPLC: ^Ct _Ret=1.46 minutes.

¹H-NMR(300MHz，CDCl ₃)：7.45(s，1H)，7.30-7.25(m，1H)，7.05-7.00(m，1H)，6.95-6.90(m，1H)，6.85-6.80(m，1H)，4.00(s/broad，2H)，3.30-3.20(m，4H)，2.65-2.58(m，4H)，2.35(s，3H)。

Stages 25.1 2-(1-Methyl-1H-indole-3-yl)-3-oxo-propionitrile

With the preparation of the chemical compound in stage 1.3 Synthetic 2-(1-Methyl-1H-indole-3-yl)-3-oxo-propionitrile similarly: productive rate: 59%; Oily; ES-MS:M+H=199.1.

Chemical compound forms tautomeric equilibrium in solution: ¹H-NMR (300MHz, CDCl ₃): 8.00/7.95 (s/s, 1H), 7.60-7.20 (m, 5H), 3.75 (s, 3H).

Stages 26.1 2-(4-methoxyl group-phenyl)-3-oxo-propionitrile

With the preparation of the chemical compound in stage 1.3 Synthetic 2-(4-methoxyl group-phenyl)-3-oxo-propionitrile similarly: productive rate: 80%; White solid; ES-MS:M-H=174.3.

Chemical compound forms tautomeric equilibrium in solution: ¹H-NMR (300MHz, DMSO-d ₆): 7.80/7.58 (s/s, 1H), 7.55-7.50 (m, 1H), 7.30-7.20 (m, 1H), 6.90-6.80 (m, 2H), 3.73/3.70 (s/s, 3H).

Stages 27.1 2-(2-methoxyl group-phenyl)-3-oxo-propionitrile

With the preparation of the chemical compound in stage 1.3 Synthetic 2-(2-methoxyl group-phenyl)-3-oxo-propionitrile similarly: productive rate: 40%; The brown oily; ES-MS:M-H=174.3; HPLC ^Ct _Ret=2.01 minutes.

Stages 28.1 3-oxo-2-pyridin-3-yl-propionitrile

With the synthetic similarly 3-oxo of the preparation of the chemical compound in stage 1.3-2-pyridin-3-yl-propionitrile: productive rate: 71%; Brown solid; ES-MS:M+H=147.2; HPLC ^Ct _Ret=1.31 minutes.

Stages 28.2 4-pyridin-3-yl-1H-pyrazole-3-yl amine

With the synthetic similarly 4-pyridin-3-yl of the preparation of the chemical compound in stage 1.2-1H-pyrazole-3-yl amine: productive rate: 68%; Brown solid; ES-MS:M+H=161.2; HPLC ^Ct _Ret=0.50 minute.

Stage 30.1 [2-methoxyl group-5-(4-methyl-piperazine-1-yl)-phenyl]-acetonitrile

With synthetic similarly [2-methoxyl group-5-(4-methyl-piperazine-1-yl)-the phenyl]-acetonitrile of the preparation of the chemical compound in stage 1.5: productive rate: 51%; Brown solid; ES-MS:M+H=246.6; HPLC: ^Ct _Ret=1.72 minutes.

¹H-NMR(300MHz，CDCl ₃)：7.00-6.95(m，1H)，6.85-6.75(m，2H)，3.80(s，3H)，3.65(s，2H)，3.15-3.05(m，4H)，2.60-2.55(m，4H)，2.35(s，3H)。

Stages 30.2 2-[2-methoxyl group-5-(4-methyl-piperazine-1-yl)-phenyl]-3-oxo-propionitrile

With the preparation of the chemical compound in stage 1.4 Synthetic 2-[2-methoxyl group-5-(4-methyl-piperazine-1-yl)-phenyl]-3-oxo-propionitrile similarly: productive rate: 100%; Brown solid; ES-MS:M+H=274.1; HPLC: ^Ct _Ret=1.62 minutes.

Stages 30.3 4-[2-methoxyl group-5-(4-methyl-piperazine-1-yl)-phenyl]-2H-pyrazole-3-yl amine

With the synthetic similarly 4-[2-methoxyl group-5-(4-methyl-piperazine-1-yl) of the preparation of the chemical compound in stage 1.2-phenyl]-2H-pyrazole-3-yl amine: productive rate: 32%; Brown solid; ES-MS:M+H=288.2; HPLC: ^Ct _Ret=1.46 minutes.

¹H-NMR(300MHz，CDCl ₃)：7.50(s，1H)，7.00-6.95(m，1H)，6.90-6.80(m，2H)，3.80(s，3H)，3.20-3.10(m，4H)，2.65-2.55(m，4H)，2.35(s，3H)。

Stages 32.1 2-(2-benzyloxy-phenyl)-3-oxo-propionitrile

With the preparation of the chemical compound in stage 1.3 Synthetic 2-(2-benzyloxy-phenyl)-3-oxo-propionitrile similarly: productive rate: 85%; White solid; ES-MS:M+H=252.6; HPLC: ^Ct _Ret=2.35 minutes.

Chemical compound forms tautomeric equilibrium in solution: ¹H-NMR (300MHz, DMSO-d ₆): 11.6/7.78 (s, 1H), 7.55-7.45 (m, 2H), 7.40-7.20 (m, 5H), 7.15-7.05 (m, 1H), 7.00-6.90 (m, 1H), 5.15 (s, 2H).

Stages 34.1 2-(4-benzyloxy-phenyl)-3-oxo-propionitrile

With the preparation of the chemical compound in stage 1.3 Synthetic 2-(4-benzyloxy-phenyl)-3-oxo-propionitrile similarly: productive rate: 95%; White solid; ES-MS:M-H=250.3; HPLC: ^Ct _Ret=2.41 minutes.

Chemical compound forms tautomeric equilibrium in solution: ¹H-NMR (300MHz, DMSO-d ₆): 12.0/11.7 (s, 1H), 7.90-7.80 and 7.60-7.50 (m, 1H), 7.40-7.25 (m, 6H), 7.05-6.95 (m, 2H), 5.10 (s, 2H).

Stages 44.1 4-(1-Methyl-1H-indole-3-yl)-2H-pyrazole-3-yl amine

With the synthetic similarly 4-(1-Methyl-1H-indole-3-yl) of the preparation of the chemical compound in stage 1.2-2H-pyrazole-3-yl amine: productive rate: 10%; The brown foam; ES-MS:M+H=213.2; HPLC: ^Ct _Ret=1.66 minutes.

¹H-NMR(300MHz，DMSO-d ₆)：7.70(d，1H)，7.60(s，1H)，7.35(d，1H)，7.30-7.25(m，1H)，7.20-7.10(m，2H)，3.80(s，3H)。

Stages 47.1 2-(2-methoxyl group-phenyl)-3-oxo-propionitrile

With the preparation of the chemical compound in stage 1.3 Synthetic 2-(2-methoxyl group-phenyl)-3-oxo-propionitrile similarly: productive rate: 59%; White solid; ES-MS:M+H=175.3; HPLC: ^Ct _Ret=2.01 minutes.

Stages 47.2 4-(2-methoxyl group-phenyl)-2H-pyrazole-3-yl amine

With the synthetic similarly 4-(2-methoxyl group-phenyl) of the preparation of the chemical compound in stage 1.2-2H-pyrazole-3-yl amine: productive rate: 35%; White solid; ES-MS:M+H=190.1; HPLC: ^Ct _Ret=1.40 minutes.

¹H-NMR(300MHz，DMSO-d ₆)：11.5(bs，1H)，7.50(bs，1H)，7.30(bs，1H)，7.20-7.05(m，1H)，7.00-6.85(m，2H)，4.30(bs，2H)，3.75(s，3H)。

Stages 51.1 3-oxo-2-pyridin-4-yl-propionitrile

With the synthetic similarly 3-oxo of the preparation of the chemical compound in stage 1.3-2-pyridin-4-yl-propionitrile: productive rate: 59%; Orange solids; ES-MS:M+H=147.2; HPLC: ^Ct _Ret=1.00 minutes.

Chemical compound forms tautomeric equilibrium in solution: ¹H-NMR (300MHz, DMSO-d ₆): 13.1/9.60 (bs, 1H), 9.10 (bs, 1H), 8.20-8.00 (m, 2H), 7.95-7.80 (m, 1H).

Stage 52.1 (Z)-3-dimethylamino-2-(3-nitro-phenyl)-acrylonitrile

To be dissolved in dimethylbenzene (30mL) (3-nitro-phenyl)-acetonitrile (1.51g, 9.31mmol), (6.2mL 46.5mmol) reflux to stir 1 hour to dimethoxy-methyl-dimethyl-amine.Add hexane (20mL) afterwards 0 ℃ of reaction mixture.Leach deposit to obtain brown solid chemical compound (1.76g, the 8.19mmol in stage 52.1; 88%); ES-MS:M+H=218.1; HPLC: ^Ct _Ret=2.24 minutes.

¹H-NMR(300MHz，DMSO-d ₆)：8.10-8.05(m，1H)，7.90-7.85(m，1H)，7.75-7.72(m，1H)，7.70(s，1H)，7.65-7.60(m，1H)，3.30(s，6H)。

Stages 52.2 3-[3-(4-methyl-piperazine-1-yl)-phenyl]-6-(3-nitro-phenyl)-pyrazolo [1,5-a] pyrimidin-7-yl amine

To be dissolved in 4-[3-(4-methyl-piperazine-1-yl)-phenyl of AcOH (10mL) and BuOH (10mL)]-1H-pyrazole-3-yl amine (stage 24.3) (305mg, 1.18mmol), (335mg 1.54mmol) reflux to stir 16 hours to (Z)-3-dimethylamino-2-(3-nitro-phenyl)-acrylonitrile (stage 52.1).Add saturated NaHCO ₃Use EtOAc (50mL, 2x) extractive reaction mixture after the aqueous solution.Use H ₂The organic facies that O (10mL) washing merges, dry (Na ₂SO ₄), under reduced pressure, concentrate and flash chromatography (silica gel, 2.5 * 15cm, CH ₂Cl ₂/ MeOH=9: 1) to obtain orange solids chemical compound (224mg, the 0.52mmol in stage 52.2; 44%); ES-MS:M+H=430.1; HPLC: ^Ct _Ret=1.91 minutes.

¹H-NMR(300MHz，DMSO-d ₆)：8.70(s，1H)，8.35-8.30(m，1H)，8.25(s，1H)，8.22-8.18(m，1H)，7.98-7.95(m，1H)，7.90(bs，2H)，7.80-7.70(m，2H)，7.65-7.60(m，1H)，7.25-7.18(m，1H)，6.80-6.75(m，1H)，3.20-3.10(m，4H)，2.50-2.40(m，4H)，2.20(s，3H)。

Stages 53.1 3-[4-(4-methyl-piperazine-1-yl)-phenyl]-6-(3-nitro-phenyl)-pyrazolo [1,5-a] pyrimidin-7-yl amine

With the synthetic similarly 3-[4-(4-methyl-piperazine-1-yl) of the preparation of the chemical compound in stage 52.2-phenyl]-6-(3-nitro-phenyl)-pyrazolo [1,5-a] pyrimidin-7-yl amine: productive rate: 30%; Red solid; ES-MS:M+H=430.0.

¹H-NMR(300MHz，DMSO-d ₆)：8.60(s，1H)，8.35-8.30(m，1H)，8.22(s，1H)，8.20-8.10(m，1H)，8.00(d，2H，J＝7.9Hz)，7.95-7.90(m，1H)，7.85(bs，2H)，7.80-7.75(m，1H)，6.95(d，1H，J＝7.9Hz)，3.20-3.10(m，4H)，2.50-2.40(m，4H)，2.20(s，3H)。

Stage 54.1 (Z)-3-dimethylamino-2-(2-nitro-phenyl)-acrylonitrile

With synthetic similarly (the Z)-3-dimethylamino-2-(2-nitro-phenyl) of the preparation of the chemical compound in stage 52.1-acrylonitrile: productive rate: 97%; Brown solid.

¹H-NMR(300MHz，DMSO-d ₆)：7.82-7.78(m，1H)，7.62-7.55(m，1H)，7.45-7.35(m，2H)，7.20(s，1H)，3.15(s，6H)。

Stages 54.2 3-[4-(4-methyl-piperazine-1-yl)-phenyl]-6-(2-nitro-phenyl)-pyrazolo [1,5-a] pyrimidin-7-yl amine

With the synthetic similarly 3-[4-(4-methyl-piperazine-1-yl) of the preparation of the chemical compound in stage 55.2-phenyl]-6-(2-nitro-phenyl)-pyrazolo [1,5-a] pyrimidin-7-yl amine: brown solid; ES-MS:M+H=430.0; HPLC: ^Dt _Ret=1.61 minutes.

¹H-NMR(300MHz，DMSO-d ₆)：8.55(s，1H)，8.20-8.15(m，1H)，8.05-7.95(m，3H)，7.82-7.60(m，5H)，7.00-6.95(m，2H)，3.15-3.05(m，4H)，2.45-2.40(m，4H)，2.20(s，3H)。

Stage 55.1 (E)-3-dimethylamino-2-(4-methyl-thiazol-2-yl)-acrylonitrile

With synthetic similarly (the E)-3-dimethylamino-2-(4-methyl-thiazol-2-yl) of the preparation of the chemical compound in stage 52.1-acrylonitrile: productive rate: 74%; Black solid; ES-MS:M+H=194.2; HPLC: ^Ct _Ret=1.57 minutes.

¹H-NMR(300MHz，DMSO-d ₆)：7.76(s，1H)，6.60(s，1H)，3.25(bs，6H)，2.35(s，3H)。

Embodiment 61

3-{7-amino-2-methyl-3-[4-(4-methyl-piperazine-1-yl) phenyl]-pyrazolo [1,5-a] pyrimidine-6-yl }-phenol

When forming the pyrazoles ring, do not use hydrazine by using methylhydrazine, with synthetic similarly 3-{7-amino-2-methyl-3-[4-(4-methyl-piperazine-1-yl) phenyl of the preparation of the chemical compound of embodiment 1]-pyrazolo [1,5-a] pyrimidine-6-yl }-phenol: ES-MS:M+H=415.2; HPLC: ^Dt _Ret=1.45 minutes.

¹H-NMR(300MHz，DMSO-d ₆)：9.53(s，1H，OH)，2.56(s，3H CH ₃)，2.24(s，3H CH ₃)。

Embodiment 62

(4-{7-amino-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidine-6-yl }-phenyl)-ammonia

The base Ethyl formate

Under argon environment, will be dissolved in EtOH (4mL) and ethanol HCl (1.6mL; 2.5N) 4-(4-(4-methyl-piperazine-1-yl)-phenyl)-2H-pyrazole-3-yl amine (stage 1.2) (200mg; 0.81mmol) and [4-(2-cyano group-1-formoxyl-ethyl)-phenyl]-urethanes (stage 62.1) (275mg 0.04mmol) stirred 17 hours under refluxing.Add H ₂O (4mL) and K ₂CO ₃(250mg) use CH afterwards ₂Cl ₂(20mL, 2x) extractive reaction mixture.Use H ₂The organic facies that O (10mL) washing merges, dry (Na ₂SO ₄), under reduced pressure, concentrate and flash chromatography (silica gel, 2.5 * 15cm, CH ₂Cl ₂/ MeOH/NH ₃=95: 5: 0.5) to obtain white solid chemical compound (58mg, the 0.123mmol of embodiment 62; 15%); ES-MS:M+H=472.0; R _f(CH ₂Cl ₂/ MeOH/NH ₃=90: 10: 0.1)=0.42; HPLC: ^At _Ret=4.26 minutes.

¹H-NMR (400MHz, DMSO-d ₆): 8.75/8.58 (s/s, 1H/1H, pyrazolopyrimidine base), 8.03 (d, 9.0Hz, 2H, phenyl), 7.61 (d, 9Hz, 2H, phenyl), 7.53 (s, 2H, NH ₂), 7.46 (d, 9Hz, 2H, phenyl), 7.00 (d, 9Hz, 2H, phenyl), 4.17 (q, 7.5Hz, 2H, CH ₂-ethyl), 3.17/2.48 (m/m, 4H/4H, piperazinyl), 2.24 (t, 7.5Hz, 3H, CH ₃).

Stage 62a.1 [4-(cyano group-1-formoxyl-methyl)-phenyl]-urethanes

Preparation formylated similarly [4-(cyano group-methyl)-phenyl]-carbamic acid benzyl ester (stage 62a.2) (1g with the stage 1.3; 3.76mmol) to obtain corresponding urethanes (also therefore benzyl ester functional group being transformed into ethyl ester functional group): clear crystal (654mg; 2.66mmol, 70%).ES-MS：M+H＝233.0。

¹H-NMR (400MHz, DMSO-d ₆): 4.12 (q/broad, 7.5Hz, 2H, CH ₂-ethyl), 1.23 (t/broad, 7.5Hz, 3H, CH ₃-ethyl).

Stage 62.2 [4-(cyano group-methyl)-phenyl]-carbamic acid benzyl ester

Stirring at room be dissolved in two  alkane (16mL) (4-amino-phenyl)-acetonitrile (2g, 15.1mmol) and heavy carbonic dibenzyl ester (4.33g, 15.1mmol) 1 hour.Pass through flash chromatography separated product (silica gel, 4.5 * 25cm, CH behind the evaporating solvent ₂Cl ₂/ MeOH=99: 1): white solid (3.82g, 14.4mmol; 95%); ES-MS:M-H=265.0; R _f(CH ₂Cl ₂/ MeOH=95: 5)=0.49; HPLC: ^At _Ret=6.32 minutes.

¹H-NMR (400MHz, DMSO-d ₆): 9.82 (s, 1H, NH), 7.51-7.35 (m, 7H, aryl), 7.26 (d, 8.5Hz, 2H, aryl), 5.15 (s, 2H, CH ₂), 3.95 (s, 2H, CH ₂).

(Z-) 3-dimethylamino-2-thiazole-4-base-acrylonitrile

With synthetic (Z-) 3-dimethylamino-2-thiazole-4-base-acrylonitrile: ES-MS[M+1 like the compounds in stage 52.1] ⁺=180.1; HPLC: ^Ct _Ret=1.91 minutes.

Exist under the condition of pyridine by making amino precursor and corresponding sulfonic acid chloride or acetic anhydride preparation have the chemical compound 61,62,64,67 and 68 and have a chemical compound (chemical compound 63,65 and 69) of acetamide functional group of sulfonamides functional group.

Embodiment 70 and 71

According to the chemical compound in embodiment 1 preparation table 2 and the table 3.

Table 2-embodiment 70

Table 3-embodiment 71

Embodiment 72

6-(3-chloro-phenyl)-5-methyl-3-[3-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl

Amine

Under the room temperature with 4-[3-(4-methyl-piperazine-1-yl)-phenyl]-1H-pyrazole-3-yl amine (stage 72.2) (1.29g, 5mmol) be dissolved among the EtOH (25mL), add 2-(3-chloro-phenyl)-3-oxo-butyronitrile (stage 72.3) (0.97g then, 5mmol) and HCl (1.25M is dissolved in EtOH; 20mmol, 16mL).Under the stirring condition xanchromatic solution was refluxed 20 hours.Add H behind the cool to room temperature ₂O (80mL) and K ₂CO ₃(2.5g) making mixture is alkalescence.Use CH ₂Cl ₂(200mL, 2x) aqueous layer extracted.Use H ₂O (50mL, the 2x) organic facies of washing merging, dry (Na ₂SO ₄), under reduced pressure, concentrate and chromatography (silica gel, 120g RediSep, ISCO Sg-100CH ₂Cl ₂/ MeOH/NH ₃=95: 5: 0.1) to obtain title compound 72 (1.03g, the 2.38mmol of white crystal; 48%); Mp.110-115 ℃; MS (ESI+): m/z=433 (M+H) ⁺HPLC: ^At _Ret=3.72 minutes (system 1).

Stage 72.1:2-(3-chloro-phenyl)-3-oxo-butyronitrile

Under the N2 environment, 355ml ethanol is heated to 55 ℃.Added sodium (3.91g in 30 minutes in introversive this solution; 0.17mol) and stir 1.5h up to all dissolving metals.In colourless solution, add 3-chlorine benzyl cyanide (15.31g; 0.1mol) and ethyl acetate (28.53mL; 0.29mol), under refluxing, stir 5h then.The reaction finish after with yellow mixture cool to room temperature and evaporation under reduced pressure.Crude material is placed water (200mL) and passes through to add the neutralization of 25g citric acid.Use CH ₂Cl ₂(2 * 250mL) aqueous layer extracted.Use H ₂O (organic facies that 2 * 150mL) washings merge, dry (Na ₂SO ₄), concentrated also chromatography under reduced pressure (Merck 60 (0.040-0.063) is with 1: 1 eluting of EtOAc/ hexane for silica gel, 1kg) is to obtain title compound 72.1 (9.7g, the 0.05mol of light yellow crystal; 50%); Mp.92-97 ℃; MS (ESI+): m/z=302.9 (M+H) ⁺HPLC: ^At _Ret=5.67 minutes (system 1).

Stage 72.2:4-[3-(4-methyl-piperazine-1-yl)-phenyl]-1H-pyrazole-3-yl amine such as embodiment 24; The described preparation title compound of stage 24.1-24.3.

Embodiment 73

6-(3-chloro-phenyl)-5-methyl-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-[4-(4-methyl-piperazine-1-yl)-phenyl]-(embodiment 1 for 2H-pyrazole-3-yl amine; Stage 1.2) and 2-(3-chloro-phenyl)-3-oxo-butyronitrile (embodiment 73, the stage 73.1).The ecru crystal; Mp.113-115 ℃; MS (ESI+): m/z=433 (M+H) ⁺HPLC: ^At _Ret=3.56 minutes (system 1).

Embodiment 74

6-(3-chloro-phenyl)-3-[2-methoxyl group-5-(4-methyl-piperazine-1-yl)-phenyl]-5-

Methyl-pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-[2-methoxyl group-5-(4-methyl-piperazine-1-yl)-phenyl]-2H-pyrazole-3-yl amine and 2-(3-chloro-phenyl)-3-oxo-butyronitrile (embodiment 72, the stage 72.1).The ecru crystal; Mp.116-121 ℃; MS (ESI+): m/z=463 (M+H) ⁺HPLC: ^At _Ret=3.68 minutes (system 1).

Stage 74.1:4-[2-methoxyl group-5-(4-methyl-piperazine-1-yl)-phenyl]-2H-pyrazole-3-yl amine

As the embodiment 1 (stage 1.2; Stage 1.4 and 1.5) described preparation title compound, but use is 5-bromo-2-methoxyl group-phenylacetonitrile and N methyl piperazine.Weak yellow foam; MS (ESI ⁺): m/z=288.2 (M+H) ⁺HPLC: ^At _Ret=3.53 minutes (system 2).

Embodiment 75

6-(3-chloro-phenyl)-3-[2-methoxyl group-4-(4-methyl-piperazine-1-yl)-phenyl]-the 5-methyl-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-[2-methoxyl group-4-(4-methyl-piperazine-1-yl)-phenyl]-2H-pyrazole-3-yl amine and 2-(3-chloro-phenyl)-3-oxo-butyronitrile (embodiment 72, the stage 72.1).The ecru crystal; Mp.215-217 ℃; MS (ESI+): m/z=463 (M+H) ⁺HPLC: ^At _Ret=3.63 minutes (system 1).

Stage 75.1:4-[2-methoxyl group-5-(4-methyl-piperazine-1-yl)-phenyl]-2H-pyrazole-3-yl amine

As the embodiment 1 (stage 1.2; Stage 1.4 and 1.5) described preparation title compound, but use is 4-bromo-2-methoxyl group-phenylacetonitrile and N methyl piperazine.Green and brown color crystal; Mp.173.7-178.1 ℃; MS (ESI ⁺): m/z=288.1 (M+H) ⁺HPLC: ^At _Ret=3.40 minutes (system 2).

Embodiment 76

3-{7-amino-3-[2-methoxyl group-4-(4-methyl-piperazine-1-yl)-phenyl]-

Pyrazolo [1,5-a] pyrimidine-6-yl }-phenol

As described in the embodiment 1 by with 6-(3-benzyloxy-phenyl)-3-[2-methoxyl group-4-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl amine solvent carries out catalytic hydrogenation and prepares title compound in methanol and existing under the Pd/C condition.The ecru crystal; Mp.217-220 ℃; MS (ESI+): m/z=431.0 (M+H) ⁺HPLC: ^At _Ret=2.65 minutes (system 1).

Stage 76.1:6-(3-benzyloxy-phenyl)-3-[2-methoxyl group-4-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl amine

As the embodiment 1 (stage 1.2; Stage 1.4 and 1.5) described preparation title compound, but use is 4-bromo-2-methoxyl group-phenylacetonitrile and N methyl piperazine.Faint yellow solid; MS (ESI ⁺): m/z=521 (M+H) ⁺HPLC: ^At _Ret=4.38 minutes (system 1).

Embodiment 77

6-(2-chloro-phenyl)-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-[4-(4-methyl-piperazine-1-yl)-phenyl]-2H-pyrazole-3-yl amine and (Z)-2-(2-chloro-phenyl)-3-dimethylamino-acrylonitrile.Yellow solid; Mp.197-200 ℃; MS (ESI ⁺): m/z=419 (M+H) ⁺HPLC: ^At _Ret=3.33 minutes (system 1).

Stage 77.1:(Z)-2-(2-chloro-phenyl)-3-dimethylamino-acrylonitrile

In argon environment under stirring condition with N, dinethylformamide-dimethyl-acetal (9.06mL; 64.3mMol) and 2-chlorine benzyl cyanide (1.95g; 12.86mMol) be heated to 100 ℃.Behind the cool to room temperature under reduced pressure enriched mixture and by chromatography purification (ISCO Sg-100 is with 1: 1 eluting of EtOAc/ hexane for silica gel, 120g RediSep) to obtain the buttery title compound of yellow thickness (2.44g, 11.8mmol; 92%); MS (ESI+): m/z=207 (M+H) ⁺TLC (EtOAc/ hexane 1: 1) R _f=0.38.

Embodiment 78

6-(2-chloro-phenyl)-3-[3-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,

5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is (Z)-2-(2-chloro-phenyl)-3-dimethylamino-acrylonitrile (embodiment 77, the stage 77.1).Light yellow crystal; Mp.200-203 ℃; MS (ESI+): m/z=419.0 (M+H) ⁺HPLC: ^At _Ret=3.65 minutes (system 1).

Embodiment 79

6-(4-fluoro-phenyl)-5-methyl-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-[4-(4-methyl-piperazine-1-yl)-phenyl]-2H-pyrazole-3-yl amine and 2-(4-fluoro-phenyl)-3-oxo-butyronitrile.White crystal; Mp.289-291 ℃; MS (ESI ⁺): m/z=417.1 (M+H) ⁺HPLC: ^At _Ret=3.21 minutes (system 1).

Stage 79.1:2-(4-fluoro-phenyl)-3-oxo-butyronitrile

As embodiment 72, described preparation title compound of stages 72.1, but use is (4-fluoro-phenyl)-acetonitrile.The ecru crystal; Mp.77-83 ℃; MS (ESI+): m/z=176.9 (M+H) ⁺HPLC: ^At _Ret=5.15 minutes (system 1).

Embodiment 80

6-(4-fluoro-phenyl)-5-methyl-3-[3-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl

Amine

As preparation title compound as described in the embodiment 72, but what use is 2-(4-fluoro-phenyl)-3-oxo-butyronitrile (embodiment 79, the stage 79.1).White crystal; Mp.204-206 ℃; MS (ESI+): m/z=417.1 (M+H) ⁺HPLC: ^At _Ret=3.34 minutes (system 1).

Embodiment 81

6-(3-chloro-phenyl)-5-methyl-3-{3-[4-(1-methyl-piperidin-4-yl)-piperazine-1-yl]-phenyl }-pyrazolo

[1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-{3-[4-(1-methyl-piperidin-4-yl)-piperazine-1-yl]-phenyl }-2H-pyrazole-3-yl amine.The ecru crystal; Mp.180-185 ℃; MS (ESI+): m/z=516.0 (M+H) ⁺HPLC: ^At _Ret=4.96 minutes (system 1).

Stage 81.1:4-{3-[4-(1-methyl-piperidin-4-yl)-piperazine-1-yl]-phenyl }-2H-pyrazole-3-yl amine

As preparation title compound as described in the embodiment 1 (stage 1.2 and 1.4 and 1.5), but what use is (3-bromo-phenyl)-acetonitrile and 1-(1-methyl-piperidin-4-yl)-piperazine.Light yellow crystal; Mp.213-220 ℃; MS (ESI ⁺): m/z=341.18 (M+H) ⁺HPLC: ^At _Ret=3.57 minutes (system 1).

Embodiment 82

6-(3-chloro-4-fluoro-phenyl)-5-methyl-3-[3-(4-methyl-piperazine-1-yl)-phenyl] pyrazolo [1,5-a] pyrimidine

-7-base amine

As preparation title compound as described in the embodiment 72, but what use is 2-(3-chloro-4-fluoro-phenyl)-3-oxo-butyronitrile.White crystal; Mp.224-226 ℃; MS (ESI+): m/z=451 (M+H) ⁺HPLC: ^At _Ret=3.86 minutes (system 1).

Stage 82.1:2-(3-chloro-4-fluoro-phenyl)-3-oxo-butyronitrile

As embodiment 72, described preparation title compound of stages 72.1 wherein uses (3-chloro-4-fluoro-phenyl)-acetonitrile.White crystal; Mp.133-134 ℃; MS (ESI ^-): m/z=209.9 (M-H) ^-HPLC: ^At _Ret=5.79 minutes (system 1).

Embodiment 83

6-(3-chloro-4-fluoro-phenyl)-5-methyl-3-[4-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidine

-7-base amine

As preparation title compound as described in the embodiment 72, but what use is 4-[4-(4-methyl-piperazine-1-yl)-phenyl]-(3-chloro-4-fluoro-phenyl)-(embodiment 82 for 3-oxo-butyronitrile for 2H-pyrazole-3-yl amine and 2-; Stage 82.1).White crystal.Mp.264-265 ℃; MS (ESI+): m/z=451 (M+H) ⁺HPLC: ^At _Ret=3.72 minutes (system 1).

Embodiment 84

6-(3-bromo-phenyl)-5-methyl-3-[3-(4-methyl-piperazine-1-yl)-phenyl]-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As embodiment 72, described preparation title compound of stages 72.1, but use is 2-(3-bromo-phenyl)-3-oxo-butyronitrile.White crystal; Mp.107-113 ℃; MS (ESI+): m/z=477 (M+H) ⁺HPLC: ^At _Ret=4.90 minutes (system 1).

Stage 84.1:2-(3-bromo-phenyl)-3-oxo-butyronitrile

As embodiment 72, described preparation title compound of stages 72.1, but use is (3-bromo-phenyl)-acetonitrile.White crystal; Mp.96-100 ℃; MS (ESI ^-): m/z=235.9 (M-H) ^-HPLC: ^At _Ret=5.76 minutes (system 1).

Embodiment 85

6-(3-bromo-benzyl)-3-[3-(4-methyl-piperazine-1-yl)-phenyl]

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 3-(3-bromo-phenyl)-2-formoxyl-propionitrile.White crystal; Mp.170-171 ℃; MS (ESI+): m/z=477.0 (M+H) ⁺HPLC: ^At _Ret=3.84 minutes (system 1).

Stage 85.1:3-(3-bromo-phenyl)-2-formoxyl-propionitrile

Under the room temperature with 3-(3-bromophenyl) propionitrile (0.703mL; 4.66mMol) and Ethyl formate (1.499mL; 18.64mMol) be dissolved in (12.5mL) among the anhydrous THF, add then NaH (60%, in mineral oil; 670mg).Behind the room temperature 17h, add extra NaH (448mg) and Ethyl formate (0.765mL).Because this causes strong exothermic reaction, so add extra solvent (THF of 15mL).After finishing (3 days), reactant mixture is cooled to 0 ℃, handles, add 6N HCl (3mL) then and make the mixture souring with a small amount of ice cube.Add entry (50mL) and use EtOAc (3 * 100mL) extraction mixture afterwards.Water (50mL, 2x), the organic facies that merges of salt water washing, dry (Na ₂SO ₄), concentrated also chromatography under reduced pressure (ISCO Sg-100 is with 1: 1 eluting of EtOAc/ hexane for silica gel, 40g RediSep) is to obtain the buttery title compound (220mg of brown; 20%); MS (ESI-): m/z=235.9 (M-H)-; TLC EtOAc/ hexane 1: 1) Rf=0.28.

Embodiment 86

6-(3-bromo-phenyl)-3-[3-(4-methyl-piperazine-1-yl)-phenyl]-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is (Z)-2-(3-bromo-phenyl)-3-dimethylamino-acrylonitrile.White crystal; Mp.195.3-197.2 ℃; MS (ESI ⁺): m/z=463.0 (M+H) ⁺HPLC: ^At _Ret=4.05 minutes (system 1).

Stage 86.1: as embodiment 77, stages 77.1 described preparation (Z)-2-(3-bromo-phenyl)-3-dimethylamino-acrylonitrile.The golden brown crystal; Mp.102-105 ℃; MS (ESI ⁺): m/z=251.0 (M+H) ⁺HPLC: ^At _Ret=6.45 minutes (system 1).

Embodiment 87

6-(3-chloro-phenyl)-5-methyl-3-(3-morpholine-4-base-phenyl)-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-(3-morpholine-4-base-phenyl)-2H-pyrazole-3-yl amine.The canescence crystal; Mp.165-167 ℃; MS (ESI ⁺): m/z=420 (M+H) ⁺HPLC: ^At _Ret=4.49 minutes (system 1).

Stage 87.1:4-(3-morpholine-4-base-phenyl)-2H-pyrazole-3-yl amine

As embodiment 1, (stage 1.2, stage 1.4 and 1.5) described preparation title compound, but use is (3-bromo-phenyl)-acetonitrile and morpholine.The canescence crystal; Mp.166-168 ℃; MS (ESI ⁺): m/z=245.1 (M+H) ⁺HPLC: ^At _Ret=1.79 minutes (system 1).

Embodiment 88

6-(3-chloro-phenyl)-3-(4-methoxyl group-phenyl)-5-methyl-pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-(4-methoxyl group-phenyl)-2H-pyrazole-3-yl amine.White crystal; Mp.171-172 ℃; MS (ESI ⁺): m/z=365 (M+H) ⁺HPLC: ^At _Ret=4.96 minutes (system 1).

Stage 88.1:4-(4-methoxyl group-phenyl)-2H-pyrazole-3-yl amine

As embodiment 1, (stage 1.4 and 1.2) described preparation title compound, but use is (4-methoxyl group-phenyl)-acetonitrile.White crystal; Mp.198-201 ℃; MS (ESI ⁺): m/z=190 (M+H) ⁺HPLC: ^At _Ret=2.85 minutes (system 1).

Embodiment 89

6-(3-chloro-phenyl)-3-[3-((2R, 6S)-2,6-dimethyl-morpholine-4-yl)-phenyl]-5-methyl-pyrazolo

[1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but use be 4-[3-((2R, 6S)-2,6-dimethyl-morpholine-4-yl)-phenyl]-2H-pyrazole-3-yl amine.White crystal; Mp.165-167 ℃; MS (ESI ⁺): m/z=448 (M+H) ⁺HPLC: ^At _Ret=5.14 minutes (system 1).

Stage 89.1:4-[3-((2R, 6S)-2,6-dimethyl-morpholine-4-yl)-phenyl]-2H-pyrazole-3-yl amine

As embodiment 1, (stage 1.2,1.4 and 1.5) described preparation title compound, but use be (3-bromo-phenyl)-acetonitrile and (2R, 6S)-2,6-dimethyl-morpholine.White crystal; Mp.158-160 ℃; MS (ESI ⁺): m/z=273.1 (M+H) ⁺HPLC: ^At _Ret=3.02 minutes (system 1).

Embodiment 90

2-(4-{3-[7-amino-6-(3-chloro-phenyl)-5-methyl-pyrazolo [1,5-a] pyrimidin-3-yl]-phenyl }-piperazine

-1-yl)-ethanol

As preparation title compound as described in the embodiment 72, but what use is 2-{4-[3-(5-amino-1H-pyrazoles-4-yl)-phenyl]-piperazine-1-yl }-ethanol.The canescence crystal; Mp.108-116 ℃; MS (ESI ⁺): m/z=463 (M+H) ⁺HPLC: ^At _Ret=3.62 minutes (system 1).

Stage 90.1:2-{4-[3-(5-amino-1H-pyrazoles-4-yl)-phenyl]-piperazine-1-yl }-ethanol

As embodiment 1, (stage 1.2,1.4 and 1.5) described preparation title compound, but use is (3-bromo-phenyl)-acetonitrile and 2-piperazine-1-base-ethanol.Weak yellow foam; Mp.40-48 ℃; MS (ESI ⁺): m/z=288.1 (M+H) ⁺HPLC: ^At _Ret=3.45 minutes (system 1).

Embodiment 91

6-benzyl-3-[3-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is that (embodiment 23 for 2-formoxyl-3-phenyl-propionitrile; Stage 23.1).Light yellow crystal; Mp.72-75 ℃; MS (ESI+): m/z=399.1 (M+H) ⁺HPLC: ^At _Ret=3.30 minutes (system 1).

Embodiment 92

6-(3-chloro-phenyl)-3-(3,4-dimethoxy-phenyl)-5-methyl fluoride-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is that (3,4-dimethoxy-phenyl)-(embodiment 93 for 2H-pyrazole-3-yl amine for 4-; Stage 93.1) and 2-(3-chloro-phenyl)-4-fluoro-3-oxo-butyronitrile.Yellow crystals; Mp.228-230 ℃; MS (ESI+): m/z=413 (M+H) ⁺HPLC: ^At _Ret=6.65 minutes (system 1).

Stage 92.1:2-(3-chloro-phenyl)-4-fluoro-3-oxo-butyronitrile

As embodiment 72, described preparation title compound of stages 72.1, but use is the fluoro-ethyl acetate.The ecru crystal; Mp.90-96 ℃; MS (ESI ^-): m/z=209.9 (M-H) ^-HPLC: ^At _Ret=5.66 minutes (system 1).

Embodiment 93

6-(3-chloro-phenyl)-3-(3,4-dimethoxy-phenyl)-5-methyl-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-(3,4-dimethoxy-phenyl)-2H-pyrazole-3-yl amine.Pale solid; Mp.223-226 ℃; MS (ESI ⁺): m/z=395.0 (M+H) ⁺HPLC: ^At _Ret=4.69 minutes (system 1).

Stage 93.1:4-(3,4-dimethoxy-phenyl)-2H-pyrazole-3-yl amine

As embodiment 1, (stage 1.4 and 1.2) described preparation title compound, but use is (3,4-dimethoxy-phenyl)-acetonitrile.White crystal; Mp.143-146 ℃; MS (ESI ⁺): m/z=220.1 (M+H) ⁺HPLC: ^At _Ret=2.28 minutes (system 1).

Embodiment 94

6-(3-chloro-4-fluoro-phenyl)-3-(3,4-dimethoxy-phenyl)-5-methyl-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is that (3,4-dimethoxy-phenyl)-(embodiment 93 for 2H-pyrazole-3-yl amine for 4-; Stage 93.1) and 2-(3-chloro-4-fluoro-phenyl)-(embodiment 82 for 3-oxo-butyronitrile; Stage 82.1).Pale solid; Mp.235-238 ℃; MS (ESI ⁺): m/z=413.0 (M+H) ⁺HPLC: ^At _Ret=4.83 minutes (system 1).

Embodiment 95

6-(3-chloro-4-fluoro-phenyl)-3-(4-methoxyl group-phenyl)-5-methyl-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is that 4-(4-methoxyl group-phenyl)-(embodiment 88 for 2H-pyrazole-3-yl amine; Stage 88.1) and 2-(3-chloro-4-fluoro-phenyl)-(embodiment 82 for 3-oxo-butyronitrile; Stage 82.1).White crystal; Mp.224-227 ℃; MS (ESI ⁺): m/z=383 (M+H) ⁺HPLC: ^At _Ret=5.08 minutes (system 1).

Embodiment 96

6-(4-fluoro-phenyl)-3-(4-methoxyl group-phenyl)-5-methyl-pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is that (4-fluoro-phenyl)-(embodiment 79 for 3-oxo-butyronitrile for 4-(4-methoxyl group-phenyl)-2H-pyrazole-3-yl amine (embodiment 88, the stage 88.1) and 2-; Stage 79.1).White crystal; Mp.243-244 ℃; MS (ESI ⁺): m/z=349,1 (M+H) ⁺HPLC: ^At _Ret=4.56 minutes (system 1).

Embodiment 97

2-(4-{3-[7-amino-6-(4-fluoro-phenyl)-5-methyl-pyrazolo [1,5-a] pyrimidin-3-yl]-

Phenyl }-piperazine-1-yl)-ethanol

As preparation title compound as described in the embodiment 72, but what use is 2-{4-[3-(5-amino-1H-pyrazoles-4-yl)-phenyl]-piperazine-1-yl }-(4-fluoro-phenyl)-(embodiment 79 for 3-oxo-butyronitrile for ethanol (embodiment 90, the stage 90.1) and 2-; Stage 79.1).The canescence crystal; Mp.209-212 ℃; MS (ESI ⁺): m/z=447.1 (M+H) ⁺HPLC: ^At _Ret=3.24 minutes (system 1).

Embodiment 98

6-(3,4-two fluoro-phenyl)-5-methyl-3-[3-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidine

-7-base amine

As preparation title compound as described in the embodiment 72, but what use is 2-(3,4-two fluoro-phenyl)-3-oxo-butyronitrile.White solid; Mp.216-219 ℃; MS (ESI ⁺): m/z=435 (M+H) ⁺HPLC: ^At _Ret=3.30 minutes (system 1).

Stage 98.1:2-(3,4-two fluoro-phenyl)-3-oxo-butyronitrile

As embodiment 1, (stage 1.4 and 1.2) described preparation title compound, but use is (3,4-two fluoro-phenyl)-acetonitrile.White crystal; Mp.147-152 ℃; MS (ESI ⁺): m/z=195 (M+H) ⁺HPLC: ^At _Ret=5.39 minutes (system 1).

Embodiment 99

6-(3,4-two fluoro-phenyl)-3-(3,4-dimethoxy-phenyl)-5-methyl-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is that (3,4-dimethoxy-phenyl)-(embodiment 93 for 2H-pyrazole-3-yl amine for 4-; Stage 93.1) and 2-(3,4-two fluoro-phenyl)-(embodiment 98 for 3-oxo-butyronitrile; Stage 98.1).Pale solid; Mp.230-235 ℃; MS (ESI ⁺): m/z=397.0 (M+H) ⁺HPLC: ^At _Ret=4.53 minutes (system 1).

Embodiment 100

2-(4-{3-[7-amino-6-(3-chloro-4-fluoro-phenyl)-5-methyl-pyrazolo [1,5-a]

Pyrimidin-3-yl]-phenyl }-piperazine-1-yl)-ethanol

As preparation title compound as described in the embodiment 72, but what use is 2-{4-[3-(5-amino-1H-pyrazoles-4-yl)-phenyl]-piperazine-1-yl }-(3-chloro-4-fluoro-phenyl)-(embodiment 82 for 3-oxo-butyronitrile for ethanol (embodiment 90, the stage 90.1) and 2-; Stage 82.1).The canescence crystal; Mp.104-107 ℃; MS (ESI ⁺): m/z=481 (M+H) ⁺HPLC: ^At _Ret=4.00 minutes (system 1).

Embodiment 101

2-(4-{3-[7-amino-6-(3,4-two fluoro-phenyl)-5-methyl-pyrazolo [1,5-a] pyrimidin-3-yl]-phenyl }-piperazine

Piperazine-1-yl)-ethanol

As preparation title compound as described in the embodiment 72, but what use is 2-{4-[3-(5-amino-1H-pyrazoles-4-yl)-phenyl]-piperazine-1-yl }-(3,4-two fluoro-phenyl)-(embodiment 98 for 3-oxo-butyronitrile for ethanol (embodiment 90, the stage 90.1) and 2-; Stage 98.1).The canescence crystal; Mp.172-174 ℃; MS (ESI ⁺): m/z=465 (M+H) ⁺HPLC: ^At _Ret=3.71 minutes (system 1).

Embodiment 102

6-(3-chloro-phenyl)-5-methyl-3-[3-(4-pyrrolidine-1-base-piperidines-1-yl)-phenyl]-pyrazolo [1,5-a] is phonetic

Pyridine-7-base amine

As preparation title compound as described in the embodiment 72, but what use is 4-[3-(4-pyrrolidine-1-base-piperidines-1-yl)-phenyl]-1H-pyrazole-3-yl amine.Yellow crystals; Mp.188-193 ℃; MS (ESI ⁺): m/z=487.0 (M+H) ⁺HPLC: ^At _Ret=4.21 minutes (system 1).

Stage 102.1:4-[3-(4-pyrrolidine-1-base-piperidines-1-yl)-phenyl]-1H-pyrazole-3-yl amine

As embodiment 1, (stage 1.2,1.4 and 1.5) described preparation title compound, but use is (3-bromo-phenyl)-acetonitrile and 4-pyrrolidine-1-base-piperidines.Yellow crystals; Mp.214-216 ℃; MS (ESI ⁺): m/z=312.1 (M+H) ⁺HPLC: ^At _Ret=3.71 minutes (system 1).

Embodiment 103

6-(4-fluoro-phenyl)-5-methyl-3-[3-(4-pyrrolidine-1-base-piperidines-1-yl)-

Phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-[3-(4-pyrrolidine-1-base-piperidines-1-yl)-phenyl]-(embodiment 102 for 1H-pyrazole-3-yl amine; Stage 102.1) and 2-(4-fluoro-phenyl)-(embodiment 79 for 3-oxo-butyronitrile; Stage 79.1).White crystal; Mp.244-249 ℃; MS (ESI ⁺): m/z=471.0 (M+H) ⁺HPLC: ^At _Ret=3.82 minutes (system 1).

Embodiment 104

6-(3-chloro-phenyl)-3-[3-(4-lignocaine-piperidines-1-yl)-phenyl]-5-methyl-pyrazolo [1,5-a] pyrimidine

-7-base amine

As preparation title compound as described in the embodiment 72, but what use is { 1-[3-(3-amino-1H-pyrazoles-4-yl)-phenyl]-piperidin-4-yl }-diethyl-amine.White crystal; Mp.163-168 ℃; MS (ESI ⁺): m/z=489.0 (M+H) ⁺HPLC: ^At _Ret=4.02 minutes (system 1).

Stage 104.1:{1-[3-(3-amino-1H-pyrazoles-4-yl)-phenyl]-piperidin-4-yl }-diethyl-amine

As embodiment 1, (stage 1.2,1.4 and 1.5) described preparation title compound, but use is (3-bromo-phenyl)-acetonitrile and diethyl-piperidin-4-yl-amine.The ecru solid, amorphous; MS (ESI ⁺): m/z=314.2 (M+H) ⁺HPLC: ^At _Ret=3.75 minutes (system 1).

Embodiment 105

3-[3-(4-lignocaine-piperidines-1-yl)-phenyl]-6-(4-fluoro-phenyl)-5-methyl-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is that (4-fluoro-phenyl)-(embodiment 79 for 3-oxo-butyronitrile for { 1-[3-(3-amino-1H-pyrazoles-4-yl)-phenyl]-piperidin-4-yl }-diethyl-amine (embodiment 104, the stage 104.1) and 2-; Stage 79.1).White crystal; Mp.208-210 ℃; MS (ESI ⁺): m/z=473.1 (M+H) ⁺HPLC: ^At _Ret=3.63 minutes (system 1).

Embodiment 106

6-(4-fluoro-phenyl)-5-methyl-3-[3-(4-methyl-4-oxygen base-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] is phonetic

Pyridine-7-base amine

With 6-(4-fluoro-phenyl)-5-methyl-3-[3-(4-methyl-piperazine-1-yl)-phenyl]-pyrazolo [1,5-a] pyrimidin-7-yl amine (embodiment 80) (50mg; 0.12mMol) be dissolved in CH ₂Cl ₂(10mL) and at 0 ℃ with 3-chlorine benzylhydroperoxide (31.1mg; 0.126mMol) processing 1h, stirring at room 2h then.After going down to desolventize, reduced pressure passes through chromatography purification crude mixture (silica gel, 12g RediSep, ISCOSg-100 CH ₂Cl ₂/ MeOH/NH ₃=80: 20: 1) to obtain the crystalline title compound of ecru (44mg); Mp.210-223 ℃; MS (ESI ⁺): m/z=449 (M+H) ⁺HPLC: ^At _Ret=3.31 minutes (system 1).

Embodiment 107

6-(4-fluoro-phenyl)-5-methyl-3-[3-(4-methyl isophthalic acid, 4-dioxy base-piperazine-1-yl)-phenyl]-pyrazolo

[1,5-a] pyrimidin-7-yl amine

From embodiment 106 described same Reaction Separation title compounds: ecru crystal (20mg); Mp.161-169 ℃; MS (ESI+): m/z=433 (M+H)+; HPLC: ^At _Ret=3.89 minutes (system 1).

Embodiment 108

6-(3-chloro-phenyl)-3-[3-(4-dimethylamino-piperidines-1-yl)-phenyl]-5-methyl-pyrazolo [1,5-a] is phonetic

Pyridine-7-base amine

As preparation title compound as described in the embodiment 72, but what use is { 1-[3-(5-amino-1H-pyrazoles-4-yl)-phenyl]-piperidin-4-yl }-dimethyl-amine.

Stage 108.1 1-[3-(5-amino-1H-pyrazoles-4-yl)-phenyl]-piperidin-4-yl }-dimethyl-amine

As embodiment 1, (stage 1.2,1.4 and 1.5) described preparation title compound, but use is (3-bromo-phenyl)-acetonitrile and dimethyl-piperidin-4-yl-amine.

Embodiment 109

6-(3,4-two fluoro-phenyl)-3-[3-(4-dimethylamino-piperidines-1-yl)-phenyl]-5-methyl-pyrazolo

[1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is that { 1-[3-(5-amino-1H-pyrazoles-4-yl)-phenyl]-piperidin-4-yl }-(embodiment 108 for dimethyl-amine; Stage 108.1) and 2-(3,4-two fluoro-phenyl)-(embodiment 98 for 3-oxo-butyronitrile; Stage 98.1).

Embodiment 110

6-(3-chloro-phenyl)-5-methyl-3-(3,4,5-trimethoxy-phenyl)-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-(3,4,5-trimethoxy-phenyl)-2H-pyrazole-3-yl amine.

Stage 110.1:4-(3,4,5-trimethoxy-phenyl)-2H-pyrazole-3-yl amine

As embodiment 1, (stage 1.4 and 1.2) described preparation title compound, but use is (3,4,5-trimethoxy-phenyl)-acetonitrile.

Embodiment 111

6-(3,4-two fluoro-phenyl)-5-methyl-3-(3,4,5-trimethoxy-phenyl)-

Pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is that (3,4,5-trimethoxy-phenyl)-(embodiment 110 for 2H-pyrazole-3-yl amine for 4-; Stage 110.1) and 2-(3,4-two fluoro-phenyl)-(embodiment 98 for 3-oxo-butyronitrile; Stage 98.1).

Embodiment 112

6-(3-chloro-phenyl)-3-(3-methoxyl group-phenyl)-5-methyl-pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 72, but what use is 4-(3-methoxyl group-phenyl)-2H-pyrazole-3-yl amine.

Stage 112.1:4-(3-methoxyl group-phenyl)-2H-pyrazole-3-yl amine

As embodiment 1, (stage 1.4 and 1.2) described preparation title compound, but use is (3-methoxyl group-phenyl)-acetonitrile.

Embodiment 113

6-[7-amino-3-(3,4-dimethoxy-phenyl)-pyrazolo [1,5-a] pyrimidine-6-yl]-pyridine-2-alcohol

As preparation title compound as described in the embodiment 1, but what use is that (3,4-dimethoxy-phenyl)-(embodiment 93 for 2H-pyrazole-3-yl amine for 2-(6-hydroxyl-pyridine-2-yl)-3-oxo-propionitrile and 4-; Stage 96.1).

Embodiment 114

6-benzyl-3-(3,4-dimethoxy-phenyl)-pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 93, but what use is that (3,4-dimethoxy-phenyl)-(embodiment 93 for 2H-pyrazole-3-yl amine for 4-; Stage 93.1).

Embodiment 115

3-(3,4-dimethoxy-phenyl)-6-(3-fluoro-benzyl)-pyrazolo [1,5-a] pyrimidin-7-yl amine

As preparation title compound as described in the embodiment 114, but what use is 2-(3-fluoro-benzyl)-3-oxo-propionitrile.

Embodiment 116

The tablet 1 that comprises formula (I) chemical compound

Prepare the tablet of following component with conventional method, wherein tablet comprises arbitrary formula (I) chemical compound of mentioning among the embodiment 1-115 of 50mg front as active component:

Component:
		Active component	50mg
Wheaten starch	60mg
		Lactose	50mg
Colloidal silica	5mg
		Talcum	9mg
Magnesium stearate	1mg
			175mg

Preparation: active component is mixed with part wheaten starch, lactose and colloidal silica, and the mixture pressure is sifted out.Another part wheaten starch mixes in water-bath to form with the water of 5 times of amounts to be stuck with paste, and the mixture of kneading initial preparation is stuck with paste up to forming soft plastic substance with this.

It is the sieve of 3mm that dried granule was pressed width of mesh, mix with remaining corn starch, magnesium stearate and steatitic (1mm sieve) mixture that sieves in advance, and compacting forms biconvex a little tablet.

Embodiment 117

The tablet 2 that comprises formula (I) chemical compound

Prepare tablet with following component according to standard method, wherein tablet formula (I) chemical compound that comprises the arbitrary embodiment 1-115 of 100mg is as active component:

Component:
		Active component	100mg
The crystallization lactose	240mg
		Microcrystalline Cellulose	80mg
PVPPXL	20mg
		Aerosil	2mg
Magnesium stearate	5mg
			447mg

Make: active component is mixed with carrier material and pass through tablet machine (Korsch EKO, Stempeldurchmesser 10mm) and suppress.

Embodiment 118

Capsule

Prepare the capsule of following component according to standard method, wherein capsule comprises formula (I) chemical compound that provides among the arbitrary embodiment 1-115 of 100mg as active component:

Component:
		Active component	100mg
Microcrystalline Cellulose	200mg
		PVPPXL	15mg
Aerosil	2mg
		Magnesium stearate	1.5mg
	318.5mg

By blending ingredients and with their sizes of packing into is the preparation of 1 hard gelatin capsule.

Embodiment 119

Kinase inhibition by The compounds of this invention

The chemical compound of previous embodiment uses method of testing mentioned above to carry out determination of activity, and the following test-compound of its Chinese style (I) shows the following kinase whose activity shown in the his-and-hers watches 4 (" x " expression has activity to the sort of kinases)." activity " is defined as the IC to kinase inhibition as used herein ₅₀Value is 10 μ M or less than 10 μ M:

Table 4:

Embodiment	c-ab l	Flt- 3	KD R	s-S cr	RE T	c-kit	Cdk1	Ins-R
									1	x	-	x	-	x	x	-	X
2	-	-	-	-	-	-	X	-
									7	-	-	-	x	-	-	-	-

Embodiment	c-ab l	Flt- 3	KD R	s-S cr	RE T	c-kit	Cdk1	Ins-R
									16	-	-	-	-	-	-	-	X
18	-	-	-	-	-	-	X	X
									19	x	-	-	x	-	-	-	-
21	-	-	-	-	-	x	-	-
									24	-	-	-	-	-	x	-	-
27	-	-	-	x	-	-	-	-
									29	-	-	x	-	x	-	-	-
35	x	-	-	-	x	-	-	-
									37	-	-	-	-	-	-	X	-
39	-	x	-	-	x	-	-	-
									45	-	-	-	-	-	-	-	-
46	x	-	x	-	-	-	X	-
									48	x	-	-	x	x	-	-	-
49	-	x	-	-	-	-	-	-
									60	x	-	-	-	-	-	X	-
61	-	-	x	-	-	-	-	-
									61a	-	-	-	-	x	-	-	-

Claims (14)

1. The purposes of the compound of formula (I) or its pharmaceutically acceptable salt, for the treatment of protein kinase-dependent diseases:

in: R ₂ is H; substituted or unsubstituted aryl; substituted or unsubstituted heteroaryl; substituted or unsubstituted aliphatic residue; functional group; 1,5a] a substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl or substituted or unsubstituted aliphatic residue to which the pyrimidinyl ring is attached; _R can be H, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aliphatic residue, functional group, or through a linking group or atom with pyrazolo[ 1,5a] a substituted or unsubstituted aliphatic residue to which the pyrimidinyl ring is attached, At least one of _R2 or _R3 is substituted or unsubstituted aryl; substituted or unsubstituted heteroaryl; or attached to the pyrazolo[1,5a]pyrimidinyl ring through a linking group or atom substituted or unsubstituted heteroaryl or substituted or unsubstituted aryl; A is H, halogen (such as bromine), aliphatic moiety, functional group, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; and R ₁ is H, halogen or lower alkyl. 2. The use of the compound of formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof, for the treatment of protein kinase-dependent diseases: Wherein in the compound of formula (I): _R is H; lower alkyl; cycloalkyl; benzyl; benzothienyl, indolyl substituted by lower alkyl; pyridyl or thiazolyl optionally substituted by lower alkyl; unsubstituted phenyl Or phenyl substituted by one or two substituents, wherein the substituents are selected from halogen, hydroxy, alkoxy, benzyloxy, cycloalkyl, amino, acetamido, lower substituted by one or two halogen Alkylsulfonamide and benzenesulfonamide; R ₃ is H; lower alkyl optionally substituted by halogen; phenyl; pyridyl or oxazolyl; A is (a) H, halogen, benzothienyl, pyridyl, methylpiperazinylphenoxy, lower alkyl substituted indolyl, (b) unsubstituted or substituted by one or more substituents Phenyl, wherein said substituent is selected from mono-, di- or tri-lower alkoxy; di-lower alkylamino; morpholinyl optionally disubstituted by alkyl; substituted by one or more substituents Piperazinyl, wherein said substituent is selected from: lower alkyl, lower alkoxy, lower alkylpiperazinyl, pyrrolidinyl, dialkylamino and lower alkanol; and R ₁ is H. 3. The use according to claim 1 or 2, wherein the protein kinase-dependent disease is a disease dependent on c-Abl, Bcr-Abl, c-Kit, c-Raf, Flt-1, Flt-3, Her-1, KDR, PDGFR kinase, c-Src, RET receptor kinase, FGF-R1, FGF-R2, FGF-R3, FGF-R4, ephrin receptor kinases (e.g. EphB2 kinase, EphB4 kinase and related Eph kinases), Casein kinases (CK-1, CK-2, G-CK), Pak, ALK, ZAP70, Jak1, Jak2, Axl, Cdk1, cdk4, cdk5, Met, FAK, Pyk2, Syk, Insulin receptor kinase, Tie- 2 or diseases with constitutively activating mutations (activating kinases) of kinases such as Bcr-Abl, c-Kit, c-Raf, Flt-3, FGF-R3, PDGF receptor, RET, and Met and (especially abnormally high expression or activated) a kinase-dependent disease or a disease that depends on the activation of a kinase pathway, or a disease that depends on any two or more of the kinases just mentioned. 4. The use according to any one of claims 1-3, wherein the kinase-dependent disease is a disease dependent on c-abl, Flt-3, KDR, c-Src, RET, EphB4, c-kit, cdk1 , FGFR-1, c-raf, Her-1, Ins-R or Tek disease. 5. Use according to any one of claims 1-4, wherein the disease to be treated is a proliferative disease, preferably a benign or especially malignant tumor, more preferably brain, kidney, liver, adrenal gland, bladder Cancer of the breast, stomach (especially gastric tumors), ovary, colon, rectum, prostate, pancreas, lung, vagina, thyroid, sarcoma, glioblastoma, multiple myeloma, or gastrointestinal cancer, especially colon cancer Or colorectal adenomas, or neck and head tumors, epidermal hyperproliferation especially psoriasis, prostatic hyperplasia, neoplasia, especially neoplasia of epithelial character, preferably breast cancer or leukemia. 6. The use according to any one of claims 1-5, wherein the disease to be treated is a disease caused by persistent angiogenesis, such as psoriasis; Kaposi's sarcoma; restenosis, such as stent-induced restenosis; uterine Endometriosis; Crohn's disease; Hodgkin's disease; leukemia; arthritis, such as rheumatoid arthritis; hemangioma; angiofibroma; eye disease, such as diabetic retinopathy and neovascular glaucoma; kidney Diseases, such as glomerulonephritis; diabetic nephropathy; malignant nephrosclerosis; thrombotic microangiopathy syndrome; transplant rejection and glomerulopathy; fibrotic diseases, such as cirrhosis of the liver; mesangial cell proliferative disease; Arteriosclerosis; nerve tissue damage; and for inhibiting reocclusion of blood vessels after balloon catheter therapy, for blood vessel repair or after insertion of mechanical devices that keep blood vessels open such as stents, as an immunosuppressant, for scarless wound healing as an adjunct in the treatment of age spots and contact dermatitis. 7. A compound of formula (I) or a pharmaceutically acceptable salt thereof: in: _R2 is H; substituted or unsubstituted aryl; substituted or unsubstituted heteroaryl; aliphatic residue; functional group; Ring-linked substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl or aliphatic residue; _R can be H, substituted or unsubstituted aryl, heteroaryl, aliphatic residue, functional group, or aliphatic residue attached to the pyrazolo[1,5a]pyrimidinyl ring through a linking group or atom base, At least one of _R2 or _R3 is substituted or unsubstituted aryl; substituted or unsubstituted heteroaryl; or attached to the pyrazolo[1,5a]pyrimidinyl ring through a linking group or atom substituted or unsubstituted heteroaryl or substituted or unsubstituted aryl, with the proviso that R _and A are not all unsubstituted phenyl; A is H, halogen (such as bromine), aliphatic moiety, functional group, substituted or unsubstituted aryl or heteroaryl; and R ₁ is H, halogen or lower alkyl. 8. The compound according to claim 7, wherein: _R is H; lower alkyl; cycloalkyl; benzyl; benzothienyl, indolyl substituted by lower alkyl; pyridyl or thiazolyl optionally substituted by lower alkyl; unsubstituted phenyl Or phenyl substituted by one or two substituents, wherein the substituents are selected from halogen, hydroxy, alkoxy, benzyloxy, cycloalkyl, amino, acetamido, lower substituted by one or two halogen Alkylsulfonamide and benzenesulfonamide; R ₃ is H; lower alkyl optionally substituted by halogen; phenyl; pyridyl or oxazolyl; A is (a) H, halogen, benzothienyl, pyridyl, methylpiperazinylphenoxy, lower alkyl substituted indolyl, (b) unsubstituted or substituted by one or more substituents Phenyl, wherein said substituent is selected from mono-, di- or tri-lower alkoxy; di-lower alkylamino; morpholinyl optionally disubstituted by alkyl; substituted by one or more substituents Piperazinyl, wherein said substituent is selected from: lower alkyl, lower alkoxy, lower alkylpiperazinyl, pyrrolidinyl, dialkylamino and lower alkanol; R _is H; and the condition Yes _R2 and A cannot both be unsubstituted phenyl. 9. A compound selected from the group consisting of: 3-{7-amino-3-[4-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidine- 6-yl}-phenol, 6-(3-benzyloxy-phenyl)-3-[4-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5 -a]pyrimidin-7-yl}-phenol, 6-(3-methoxy-phenyl)-3-[4-(4-methyl-piperazin-1-yl)-phenyl]-pyrazole And[1,5-a]pyrimidin-7-ylamine, 6-(3,5-dimethoxy-phenyl)-3-[4-(4-methyl-piperazin-1-yl)- Phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-benzyloxy-phenyl)-3-[4-(4-methyl-piperazine-1- Base)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(4-chloro-phenyl)-3-[4-(4-methyl-piperazine-1 -yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-phenyl)-3-[4-(4-methyl-piperazine- 1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-[4-(4-methyl-piperazin-1-yl)-phenyl]-6 -Phenyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 5-methyl-3-[4-(4-methyl-piperazin-1-yl)-phenyl]-6 -Phenyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-methyl-3-[4-(4-methyl-piperazin-1-yl)-phenyl]-5 -Phenyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, N-{4-[7-amino-3-(4-dimethylamino-phenyl)-pyrazolo[1 ,5-a]pyrimidin-6-yl]-phenyl}-2,3-dichloro-benzenesulfonamide, 4-chloro-benzenesulfonic acid 4-[7-amino-3-(4-dimethylamino -phenyl)-pyrazolo[1,5-a]pyrimidin-6-yl]-phenyl ester, 6-(4-methoxy-phenyl)-5-methyl-3-phenyl-pyridine Azolo[1,5-a]pyrimidin-7-ylamine, 3-(4-methoxy-phenyl)-5-methyl-6-phenyl-pyrazolo[1,5-a]pyrimidine -7-ylamine, 6-(4-bromo-phenyl)-3-(4-methoxy-phenyl)-5-methyl-pyrazolo[1,5-a]pyrimidin-7-yl Amine, 6-(4-bromo-phenyl)-5-methyl-3-phenyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(2,6-dichloro- Phenyl)-3-phenyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-(3-methoxy-phenyl)-6-phenyl-pyrazolo[1, 5-a]pyrimidin-7-ylamine, 3-bromo-5-phenyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-benzo[b]thiophen-3-yl- 3-[4-(4-Methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-(4-bromo-phenyl) -5-Phenyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-[4-(4-methyl-piperazin-1-yl)-phenyl]-6-thiophene- 3-yl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-benzo[b]thiophen-3-yl-6-(3-methoxy-phenyl)-pyrazolo [1,5-a]pyrimidin-7-ylamine, 6-benzo-3-[4-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5- a] pyrimidin-7-ylamine, 6-(3-methoxy-phenyl)-3-[3-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1 , 5-a] pyrimidin-7-ylamine, 6-(1-methyl-1H-indol-3-yl)-3-[4-(4-methyl-piperazin-1-yl)-benzene Base]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(4-methoxy-phenyl)-3-[4-(4-methyl-piperazin-1-yl )-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(2-methoxy-phenyl)-3-[4-(4-methyl-piperazine- 1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-methoxy-phenyl)-3-pyridin-3-yl-pyrazolo [1,5-a]pyrimidin-7-ylamine, 3-{7-amino-3-[3-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1, 5-a]pyrimidin-6-yl}-phenol, 6-(3-benzyloxy-phenyl)-3-[2-methoxy-5-(4-methyl-piperazin-1-yl) -Phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-{7-amino-3-[2-methoxy-5-(4-methyl-piperazine-1 -yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-6-yl}-phenol, 6-(2-benzyloxy-phenyl)-3-[4-(4-methyl -piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 2-{7-amino-3-[4-(4-methyl-piperazine -1-yl)-phenyl]-pyrazolo[1,5-a]}pyrimidin-6-yl}-phenol, 6-(4-benzyloxy-phenyl)-3-[4-(4 -Methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 4-{7-amino-3-[4-(4-methyl -piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-6-yl}-phenol, 6-(2-benzyloxy-phenyl)-3-[3- (4-Methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 2-{7-amino-3-[3-(4- Methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-6-yl}-phenol, 6-(4-benzyloxy-phenyl)-3-[ 3-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 4-{7-amino-3-[3-( 4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-6-yl}-phenol, 6-(2-benzyloxy-phenyl)-3 -[2-Methoxy-5-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 2-{7- Amino-3-[2-methoxy-5-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-6-yl}-phenol, 6-(4-Benzyloxy-phenyl)-3-[2-methoxy-5-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5- a] pyrimidin-7-ylamine, 4-{7-amino-3-[2-methoxy-5-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1 , 5-a]pyrimidin-6-yl}-phenol, 6-(3-benzyloxy-phenyl)-3-[1-methyl-1H-indol-3-yl]-pyrazolo[1 , 5-a] pyrimidin-7-ylamine, 3-[7-amino-3-(1-methyl-1H-indol-3-yl)-pyrazolo[1,5-a]pyrimidine-6 -yl]-phenol, 3-[7-amino-3-pyridin-3-yl-pyrazolo[1,5-a]pyrimidin-6-yl]-phenol, 6-(3-benzyloxy-benzene Base)-3-(2-methoxy-phenyl)-pyrazol[1,5-a]pyrimidin-7-ylamine, 3-[7-amino-3-(2-methoxy-phenyl )-pyrazolo[1,5-a]pyrimidin-6-yl]-phenol, 3-[3-(4-methyl-piperazin-1-yl)-phenyl]-6-thiophene-3- Base-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-(2-methoxy-5-(4-methyl-piperazin-1-yl)-phenyl)-6- Thiophen-3-yl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-[4-(4-methyl-piperazin-1-yl)-phenyl]-6-pyridine- 4-yl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-amino-phenyl)-3-[3-(4-methyl-piperazin-1-yl) -Phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-amino-phenyl)-3-[4-(4-methyl-piperazin-1-yl )-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(2-amino-phenyl)-3-[4-(4-methyl-piperazine-1- Base)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-[4-(4-methyl-piperazin-1-yl)-phenyl]-6-( 4-Methyl-thiazol-2-yl)-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-benzo[b]thiophen-3-yl-3-[2-methoxy -5-(4-Methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-benzo[b]thiophen-3-yl -3-[4-methoxy-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-(3-methoxy-phenyl)-6-thiophene-3- Base-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-benzyloxy-phenyl)-3-(3-methoxy-phenyl)-pyrazolo[1 , 5-a] pyrimidin-7-ylamine, 3-[7-amino-3-(3-methoxy-phenyl)-pyrazolo[1,5-a]pyrimidin-6-yl]-phenol , (4-{7-amino-3-[4-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-6-yl}-benzene Base)-ethyl carbamate, 6-(3-chloro-phenyl)-5-methyl-3-[3-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo [1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-phenyl)-5-methyl-3-[4-(4-methyl-piperazin-1-yl)-benzene Base]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-phenyl)-3-[2-methoxy-5-(4-methyl-piperazine -1-yl)-phenyl]-5-methyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-phenyl)-3-[2-methoxy Base-4-(4-methyl-piperazin-l-yl)-phenyl]-5-methyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-{7-amino -3-[2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-6-yl}-phenol, 6 -(2-chloro-phenyl)-3-[4-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(2-Chloro-phenyl)-3-[3-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine , 6-(4-fluoro-phenyl)-5-methyl-3-[4-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a] Pyrimidin-7-amine, 6-(4-fluoro-phenyl)-5-methyl-3-[3-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1 , 5-a] pyrimidin-7-ylamine, 6-(3-chloro-phenyl)-5-methyl-3-{3-[4-(1-methyl-piperidin-4-yl)- Piperazin-1-yl]-phenyl}-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-4-fluoro-phenyl)-5-methyl-3 -[3-(4-Methyl-piperazin-1-yl)-phenyl]pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-4-fluoro-benzene Base)-5-methyl-3-[4-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6- (3-Bromo-phenyl)-5-methyl-3-[3-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidine-7 -ylamine, 6-(3-bromo-benzyl)-3-[3-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidine- 7-ylamine, 6-(3-bromo-phenyl)-3-[3-(4-methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidine -7-ylamine, 6-(3-chloro-phenyl)-5-methyl-3-(3-morpholin-4-yl-phenyl)-pyrazolo[1,5-a]pyrimidine- 7-ylamine, 6-(3-chloro-phenyl)-3-(4-methoxy-phenyl)-5-methyl-pyrazolo[1,5-a]pyrimidin-7-ylamine , 6-(3-chloro-phenyl)-3-[3-((2R,6S)-2,6-dimethyl-morpholin-4-yl)-phenyl]-5-methyl-pyridine Azolo[1,5-a]pyrimidin-7-ylamine, 2-(4-{3-[7-amino-6-(3-chloro-phenyl)-5-methyl-pyrazolo[1 , 5-a] pyrimidin-3-yl]-phenyl}-piperazin-1-yl)-ethanol, 6-benzyl-3-[3-(4-methyl-piperazin-1-yl)- Phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-phenyl)-3-(3,4-dimethoxy-phenyl)-5- Fluoromethyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-phenyl)-3-(3,4-dimethoxy-phenyl)-5- Methyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-4-fluoro-phenyl)-3-(3,4-dimethoxy-phenyl) -5-methyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-4-fluoro-phenyl)-3-(4-methoxy-phenyl) -5-Methyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(4-fluoro-phenyl)-3-(4-methoxy-phenyl)-5-methyl Base-pyrazolo[1,5-a]pyrimidin-7-ylamine, 2-(4-{3-[7-amino-6-(4-fluoro-phenyl)-5-methyl-pyrazole And[1,5-a]pyrimidin-3-yl]-phenyl}-piperazin-1-yl)-ethanol, 6-(3,4-difluoro-phenyl)-5-methyl-3- [3-(4-Methyl-piperazin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3,4-difluoro-phenyl )-3-(3,4-dimethoxy-phenyl)-5-methyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 2-(4-{3-[7 -Amino-6-(3-chloro-4-fluoro-phenyl)-5-methyl-pyrazolo[1,5-a]pyrimidin-3-yl]-phenyl}-piperazin-1-yl )-ethanol, 2-(4-{3-[7-amino-6-(3,4-difluoro-phenyl)-5-methyl-pyrazolo[1,5-a]pyrimidine-3- Base]-phenyl}-piperazin-1-yl)-ethanol, 6-(3-chloro-phenyl)-5-methyl-3-[3-(4-pyrrolidin-1-yl-piperidine -1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(4-fluoro-phenyl)-5-methyl-3-[3-(4 -pyrrolidin-1-yl-piperidin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-phenyl)-3- [3-(4-Diethylamino-piperidin-1-yl)-phenyl]-5-methyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 3-[3- (4-Diethylamino-piperidin-1-yl)-phenyl]-6-(4-fluoro-phenyl)-5-methyl-pyrazolo[1,5-a]pyrimidine-7- Baseamine, 6-(4-fluoro-phenyl)-5-methyl-3-[3-(4-methyl-4-oxy-piperazin-1-yl)-phenyl]-pyrazolo [1,5-a]pyrimidin-7-ylamine, 6-(4-fluoro-phenyl)-5-methyl-3-[3-(4-methyl-1,4-dioxy-piper Oxyzin-1-yl)-phenyl]-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3-chloro-phenyl)-3-[3-(4-dimethyl Amino-piperidin-1-yl)-phenyl]-5-methyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3,4-difluoro-phenyl)- 3-[3-(4-Dimethylamino-piperidin-1-yl)-phenyl]-5-methyl-pyrazol[1,5-a]pyrimidin-7-ylamine, 6-(3 -Chloro-phenyl)-5-methyl-3-(3,4,5-trimethoxy-phenyl)-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-(3 , 4-difluoro-phenyl)-5-methyl-3-(3,4,5-trimethoxy-phenyl)-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6 -(3-chloro-phenyl)-3-(3-methoxy-phenyl)-5-methyl-pyrazolo[1,5-a]pyrimidin-7-ylamine, 6-[7- Amino-3-(3,4-dimethoxy-phenyl)-pyrazolo[1,5-a]pyrimidin-6-yl]-pyridin-2-ol, 6-benzyl-3-(3 , 4-dimethoxy-phenyl)-pyrazolo[1,5-a]pyrimidin-7-ylamine and 3-(3,4-dimethoxy-phenyl)-6-(3- Fluoro-benzyl)-pyrazolo[1,5-a]pyrimidin-7-ylamine. 10. Use of a compound according to claim 7, 8 or 9 for the preparation of a pharmaceutical composition. 11. A pharmaceutical composition comprising a compound according to claim 7, 8 or 9. 12. A pharmaceutical composition comprising a compound according to claim 7, 8 or 9 and a pharmaceutically acceptable carrier. 13. Use of a compound according to claim 1 or 2 for the preparation of a pharmaceutical composition for the treatment of kinase-dependent diseases. 14. A process for preparing a compound according to claim 7, 8 or 9, comprising: (a) reacting the nitrile A—CH ₂ —C≡N with ethyl formate in the presence of an organic solvent to form a substituted 3-oxo-propionitrile, (b) Condensing the substituted 3-oxo-propionitrile of step (a) with hydrazine monohydrate in an organic solvent to form a 2H-pyrazol-3-ylamine of formula (III): (d) formylation of substituted nitriles in the presence of ethanolate and ethyl formate to prepare 3-oxo-propionitriles of formula (II):

(c) Condensing a 3-oxo-propionitrile of formula (II) and a 2H-pyrazol-3-ylamine of formula (III) in the presence of an organic solvent to form a compound of formula (I).