CN105732615B

CN105732615B - Cdk kinase inhibitors

Info

Publication number: CN105732615B
Application number: CN201510890859.9A
Authority: CN
Inventors: 吴永谦; 王统建
Original assignee: Shandong Xuanzhu Pharma Co Ltd
Current assignee: Xuanzhu Biopharmaceutical Co Ltd
Priority date: 2014-12-31
Filing date: 2015-12-04
Publication date: 2018-05-01
Anticipated expiration: 2035-12-04
Also published as: CN105732615A

Abstract

The invention belongs to pharmaceutical technology field, and in particular to CDK kinase inhibitors, its pharmaceutically acceptable salt, its ester, its solvate and their stereoisomer shown in general formula (I), wherein R¹、R²、R³、R⁴、A¹、A²、A³, n is defined as in the description.The invention further relates to the preparation method of these compounds, pharmaceutical preparation and pharmaceutical composition containing these compounds, and the compound, its pharmaceutically acceptable salt, its ester, its solvate and their stereoisomer are preparing treatment and/or are preventing by the application in the medicine of cancer-related diseases kinase mediated CDK.

Description

CDK kinase inhibitors

Technical field

The invention belongs to pharmaceutical technology field, and in particular to CDK kinase inhibitors, its pharmaceutically acceptable salt, its ester, Its solvate and their stereoisomer, pharmaceutical preparation and pharmaceutical composition containing these compounds, and the change Compound, its pharmaceutically acceptable salt, its ester, its solvate and their stereoisomer, in preparation treatment and/or in advance Prevent by the application in the medicine of cancer-related diseases kinase mediated CDK.

Background technology

The generation of tumour is unbalance related with a variety of oncogenes and tumor suppressor gene.Nearly all oncogene, tumor suppressor gene Functional effect, finally can all converge on the cell cycle.Therefore, it can be said that tumour is a kind of cell cycle diseases (Cell Cycle Disease, CCD), it is one of approach for treating tumour to adjust or block the cell cycle.At present, it has been found that and cell The related molecule of cycle regulating is very much, wherein cell cycle protein dependent kinase (Cyclin-Dependent-Kinases, CDKs) be cell cycle regulating network core element.CDKs is catalytic subunit, is a kind of serine (Ser)/threonine (Thr) kinases, as intracellular important signal transduction molecule, participates in the different times of cell cycle.Research shows, with CDKs Centered on cell cycle regulating network, the exception of any link will all cause the cell cycle abnormal, and ultimately result in the hair of tumour It is raw.There are 21 hypotypes in CDK families at present, are made by combining to play with its modulability subunit cyclins (cyclin) With.The function of the various hypotypes of CDK, in addition to acting on the cell cycle, further includes to transcription, DNA reparations, differentiation and cell journey The adjusting of sequence death.Key effect based on CDKs played in the propagation of modulate tumor cell and death, CDKs kinases man Race provides chance and new field for the discovery of antitumor drug with development.

In medicine is researched and developed, using flavopiridol, UCN-01 etc. for representative first generation CDK inhibitor as " pan- CDK " inhibitor, they block all hypotypes of CDK families to show the high poison of comparison in clinical test in a manner of equivalent Property, what is had cannot reach effective therapeutic dose, therefore excite people to start to research and develop selective CDK inhibitor, it is desirable to improve The selectivity for the treatment of and prevent normal cell from being damaged be subject to some side effects.

In the CDK hypotypes for participating in the cell cycle, CDK4/6 plays irreplaceable effect.The cell related with cancer Period discontinuity was primarily present in G1 phases and G1/S phase conversion processes, and CDK4/6 and CyclinD combines to form kinase activity Compound, by study of tumor suppressor genes Rb product pRb phosphorylations, discharges the transcription factor E2F of combination, starts the gene related with the S phases Transcription, promotes cell by check point, and shift from the G1 phases to the S phases.The specific activation of CDK4/6 and the propagation of some tumours It is closely related, there is the exception of cyclin D-CDK4/6-INK4-Rb paths in about 80% human tumor.This path changes Become, accelerate G1 phase processes so that tumor cell proliferation is accelerated and obtains survival advantage.Therefore, the intervention to the path becomes A kind of therapeutic strategy, CDK4/6 become a kind of new anti-tumor target.CDK4/6 is as the advantage of anti-tumor target：(1) The cell of most of propagation relies on CDK2 or CDK4/6 propagation, but the inhibitor of CDK4/6 does not show " pan-CDK suppression The cytotoxicity of agent ", as bone marrow suppression and enteron aisle are reacted.(2) preclinical laboratory shows, if the horizontal rises of cell cyclin D Or P16INK4a inactivations, by increasing capacitance it is possible to increase cell is to the sensitiveness of medicine, and there are upper relative to normal cell due to tumour cell Phenomenon is stated, so adds somewhat to the targeting of medicine.

CDK9 is a kind of serine/threonine kinases as other CDK, is found in Human cDNA Library screens earliest, It combines to form positive transcriptional elongation factor b with corresponding cyclin (s) (Positivetranseriptionelongationfaetorb, p-TEFb).The compound being capable of phosphorylation rna plymerase ii (RNApolymeraseII) and some negative transcription elongation factors are so that transcription is extended from starting position.CDK9/ The unconventionality expression of cyclinT has substantial connection with many diseases.cyclinT₁/ CDK9 and estrogen receptor positive mammary gland Cancer cell, estrogen can lower the expression of HEXIMI in this kind of cell, and HEXIMI is also referred to as estrogen down-regulated gene (EDGI), HEXIMI/EDG1 can be combined and and cyclinT with estrogen receptor₁With reference to.It is overexpressed in mammary glandular cell It can suppress cell Proliferation and growth during HEXIMI/EDGI.It can suppress P-TEFb's in cell in view of HEXIMI at the same time Kinase activity, and hexa-methylene diethyl acid amide (HMBA) induced expression can be utilized in a variety of neoplastic cells, into the cell cyclinT₁/ CDK9 activity levels have impact on the one-tenth knurl ability of tumour cell.In lymphoma cell, in T cell periodic activation It was found that cyclinT₁/ CDK9 contents raise, and cyclinT₁The up-regulation of/CDK9 expression influences the activation of this kind of cell with dividing Change.Except the function in terms of tumour, cyclinT₁/ CDK9 formed compound be HIV-1Tat albumen major target class, Tat Can be with the CyclinT of P-TEFb₁Subunit with reference to and form Tat-cyclinT₁- CDK9 compounds, pass through CDK9 kinase activity phosphorus RNApolymeraseII and N-TEF is acidified, is accomplished the transcription of HIV-1.Pharmaceutical research finds the inhibition of CDK9 Compound can effectively suppress the transcription and replication of HIV-1.In addition CDK9 has in anti-inflammatory, analgesia, myocardial hypertrophy etc. Significant biological function.

Also listed so far without CDK inhibitor medicaments, including Pfizer, gift comes public with some medicine including Novartis etc. Department reports a series of preferable CDK4/6 inhibitor of selectivity successively, just in clinical experimental stage.Wherein, it is of specific interest Be Pfizer exploitation PD0332991 (palbociclib) and Eli Lilly LY2835219 (Phase III) and The LEE-011 (Phase III) of Novartis.

In April, 2013, the PD0332991 of Pfizer obtain FDA important breakthrough medicine identifications, and in August, 2014, carries to FDA New Drug Application (NDA) is handed over, seeks to ratify the bent azoles (letrozole) of PD0332991 (palbociclib) joints for not connecing previously Systematic treating has been received to control the postmenopausal women estrogen receptor positive (ER+) of the late period state of an illness, human epidermal growth factor receptor 2 The treatment of negative (HER2-) Locally Advanced or metastatic breast cancer.This exploitation to CDK4/6 inhibitor plays very positive Effect.

In order to achieve the purpose that more preferable oncotherapy effect, the market demand is better met, it is therefore desirable to be able to develop Go out the CDK kinase inhibitors of the high-efficiency low-toxicity of a new generation.The present invention will provide a kind of selective CDK kinase inhibitions of new structure Agent, and find that the compound of such structure has good drug effect, and can be CDK kinase inhibitors effectively by blood-brain barrier Treatment as the cancer of the brain provides possibility.

The content of the invention

The object of the present invention is to provide one kind to target CDK kinase inhibitors, and specific technical solution is as follows：

1st, the compound shown in general formula (I), its pharmaceutically acceptable salt, its ester, its solvate and theirs is vertical Body isomers,

Wherein

A¹、A²And A³It is independently selected from nitrogen or carbon；

R¹Selected from C_1-6Alkyl, C_1-6Alkoxy or optionally by Q¹3~8 yuan of cycloalkyl of substitution, Q¹Selected from C_1-6Alkyl, C_1-6 Alkoxy；

R²Selected from C_1-6Alkyl, C_1-6Alkoxy, cyano group, carbamoyl or C_1-6Alkyl-carbonyl-amino；

R⁴Selected from halogen or hydrogen；

R³Selected from optionally by Q²Substitution 3~8 circle heterocycles bases, 6~14 yuan of condensed hetero ring bases, 5~8 unit's heteroaryls, 6~14 yuan Thick heteroaryl, phenyl, naphthyl, 6~12 yuan of bridge heterocyclic radicals or 6~12 yuan of spiro heterocyclic radicals；" 5~8 unit's heteroaryl " is preferably " 5~7 unit's heteroaryl ", more preferably " 5~6 unit's heteroaryl ", more preferably " 5~6 membered nitrogen-containing heteroaryl base " etc., described " 6 ~14 yuan of thick heteroaryls " are preferably " 6~10 yuan of thick heteroaryls ", more preferably " 7~10 yuan of thick heteroaryls ", more preferably " 9~ 10 yuan of thick heteroaryls ", more preferably " 9~10 yuan of nitrogenous thick heteroaryls " etc.；

Q²Selected from amino, hydroxyl, halogen, trifluoromethyl, cyano group, C_1-6Alkyl, C_1-6Alkoxy, C_1-6Alkyl sulphonyl, C_1-6Alkyl sulfonyl amino, 3~8 circle heterocycles bases or 6~9 yuan of bridge heterocyclic radicals；

N is selected from 0~3 integer.

2nd, the compound as described in scheme 1, its pharmaceutically acceptable salt, its ester, its solvate and theirs is vertical Body isomers, wherein

A¹、A²And A³It is independently selected from nitrogen or carbon；

R¹Selected from C_1-4Alkyl or C_1-4Alkoxy；

R²Selected from C_1-4Alkyl, C_1-4Alkoxy, cyano group, carbamoyl or C_1-4Alkyl-carbonyl-amino；

R⁴Selected from halogen；

R³Selected from optionally by Q²5~7 circle heterocycles bases of substitution, 6~11 yuan of condensed hetero ring bases, 6~11 yuan of bridge heterocyclic radicals or 6~ 11 yuan of spiro heterocyclic radicals；" 6~11 yuan of condensed hetero ring bases " is preferably " 6~10 yuan of condensed hetero ring bases ", and more preferably " 7~10 yuan thick Heterocyclic radical ", more preferably " 9~10 yuan of condensed hetero ring bases ", more preferably " 9~10 yuan of nitrogenous condensed hetero ring bases " etc., described " 6 ~11 yuan of bridge heterocyclic radicals " are preferably " 6~9 yuan of bridge heterocyclic radicals ", more preferably " 7~10 yuan of bridge heterocyclic radicals ", more preferably " 7~9 First bridge heterocyclic radical ", more preferably " 7~8 yuan of bridge heterocyclic radicals ", more preferably " 8 yuan of bridge heterocyclic radicals ", more preferably " 8 yuan contain Nitrogen bridge heterocyclic radical " etc., " 6~11 yuan of spiro heterocyclic radicals " are preferably " 7~11 yuan of spiro heterocyclic radicals ", more preferably " 7~10 yuan of spiral shells Heterocyclic radical ", more preferably " 7~9 yuan of spiro heterocyclic radicals ", more preferably " 7~8 yuan of spiro heterocyclic radicals ", more preferably " 7~8 yuan Nitrogenous spiro heterocyclic radical " etc.；

Q²Selected from amino, hydroxyl, trifluoromethyl, cyano group, C_1-4Alkyl, C_1-4Alkoxy, C_1-4Alkyl sulphonyl, C_1-4Alkyl Sulfonamido, 5~6 circle heterocycles bases or 7~9 yuan of bridge heterocyclic radicals；" 5~6 circle heterocycles base " is preferably " 5~6 member heterocyclic ring containing nitrogens Base ", " 7~9 yuan of bridge heterocyclic radicals ", is preferably " 7~8 yuan of bridge heterocyclic radicals ", more preferably " 8 yuan of bridge heterocyclic radicals ", further Preferably " 8 yuan of nitrogenous bridge heterocyclic radicals " etc.；

N is selected from 0~3 integer.

3rd, the compound as described in scheme 2, its pharmaceutically acceptable salt, its ester, its solvate and theirs is vertical Body isomers, wherein

A¹、A²And A³It is independently selected from nitrogen or carbon；

R¹It is isopropyl；

R²Selected from methyl, methoxyl group, cyano group, carbamoyl or acetylamino；

R⁴It is fluorine；

R³Selected from optionally by Q²5~6 circle heterocycles bases of substitution, Q²Selected from amino, hydroxyl, trifluoromethyl, cyano group, C_1-4Alkane Base, C_1-4Alkoxy, C_1-4Alkyl sulphonyl, C_1-4Alkyl sulfonyl amino, 5~6 circle heterocycles bases or 7~9 yuan of bridge heterocyclic radicals；It is described " 5~6 circle heterocycles base " is preferably " 5~6 member heterocyclic ring containing nitrogen base ", " 7~9 yuan of bridge heterocyclic radicals ", is preferably that " 7~8 yuan of bridges are miscellaneous Ring group ", more preferably " 8 yuan of bridge heterocyclic radicals ", more preferably " 8 yuan of nitrogenous bridge heterocyclic radicals " etc.；

N is selected from 0~2 integer.

4th, the compound as described in scheme 3, its pharmaceutically acceptable salt, its ester, its solvate and theirs is vertical Body isomers, wherein

R³Selected from optionally by Q²5~6 member heterocyclic ring containing nitrogen bases of substitution, Q²Selected from amino, hydroxyl, trifluoromethyl, cyano group, C_1-4 Alkyl or C_1-4Alkoxy, " 5~6 member heterocyclic ring containing nitrogen base " are preferably " 6 member heterocyclic ring containing nitrogen base "；

N is 1.

5th, the compound as described in scheme 4, its pharmaceutically acceptable salt, its ester, its solvate and theirs is vertical Body isomers, wherein

R³Selected from optionally by Q²Substituted pyrrolidinyl, piperidyl or piperazinyl, Q²Selected from trifluoromethyl, C_1-4Alkyl or C_1-4Alkoxy；

N is 1.

The part of compounds of the present invention

Detailed description of the invention

" halogen atom " of the present invention includes fluorine atom, chlorine atom, bromine atoms and iodine atom.

" C of the present invention_1-6Alkyl " can be straight or branched, including such as " C_1-4Alkyl ", " C_1-3Alkyl " Deng instantiation includes but not limited to：Methyl, ethyl, n-propyl, isopropyl, normal-butyl, 2- methyl-propyls, 1- methyl-propyls, 1,1- dimethyl ethyls, n-pentyl, 3- methyl butyls, 2- methyl butyls, 1- methyl butyls, 1- ethyl propyls, n-hexyl, 4- first Base amyl group, 3- methyl amyls, 2- methyl amyls, 1- methyl amyls, 3,3- dimethylbutyls, 2,2- dimethylbutyls, 1,1- bis- Methyl butyl, 1,2- dimethylbutyls, 1,3- dimethylbutyls, 2,3- dimethylbutyls, 2- ethyl-butyls, 1,2- dimethyl propylenes Base etc..

" C of the present invention_1-6Alkoxy, C_1-6Alkyl-carbonyl-amino, C_1-6Alkyl sulphonyl, C_1-6Alkyl sulfonyl amino " Refer to C_1-6Alkyl-O-, C_1-6Alkyl-C (O) NH-, C_1-6Alkyl-SO₂-、C_1-6Alkyl-SO₂The group that NH- modes are formed, its In " C_1-6Described in alkyl " text as defined above.

" C of the present invention_1-4Alkoxy, C_1-4Alkyl-carbonyl-amino, C_1-4Alkyl sulphonyl, C_1-4Alkyl sulfonyl amino " Refer to C_1-4Alkyl-O-, C_1-4Alkyl-C (O) NH-, C_1-4Alkyl-SO₂-、C_1-4Alkyl-SO₂The group that NH- modes are formed, its In " C_1-4Described in alkyl " text as defined above.

" 3~8 yuan of cycloalkyl " of the present invention, refers to that the paraffin section of 3~8 carbon atoms removes a hydrogen atom and spreads out Raw cyclic alkyl, including such as " 3~6 yuan of cycloalkyl ", " 4~6 yuan of cycloalkyl " etc..The example includes but not limited to：Ring third Alkyl, cyclobutane base, pentamethylene base, cyclohexyl, cycloheptyl alkyl, cyclooctane base etc..

" 3~8 circle heterocycles base " of the present invention, including for example " 3~7 circle heterocycles base ", " 3~6 circle heterocycles base ", " 4~ 7 circle heterocycles bases ", " 4~6 circle heterocycles base ", " 5~7 circle heterocycles base ", " 5~6 circle heterocycles base ", " 5~6 member heterocyclic ring containing nitrogen base ", " 6 Member heterocyclic ring containing nitrogen base " etc..Instantiation includes but are not limited to：Aziridine base, 2H- aziridine base, diazacyclo Propyl, 3H- diazacyclos acrylic, azetidinyl, 1,4- dioxanes base, 1,3- dioxanes base, 1,3- dioxolane base, 1,4- Dioxins base, tetrahydrofuran base, pyrrolin base, pyrrolidinyl, imidazolidine Base, 4,5- glyoxalidine base, pyrazolidinyl, 4,5- pyrazolines base, 2,5- dihydro-thiophenes base, tetrahydro-thienyl, 4,5- dihydros Thiazolyl, piperidyl, piperazinyl, morpholinyl base, 4,5- dihydro-oxazoles base, 4,5- dihydro-isoxazoles base, 2,3- dihydro-isoxazoles Base, 2H-1,2- oxazinyls, 6H-1,3- oxazinyls, 4H-1,3- thiazinyls, 6H-1,3- thiazinyls, 2H- pyranoses, 2H- pyrans- 2- ketone groups, 3,4- dihydro -2H- pyranoses etc., are preferably " 5~6 member heterocyclic ring containing nitrogen base ".

According to the rule of IUPAC Compound nomenclatures, condensed ring of the present invention refers to by two or more ring-type knots Structure shares two adjacent atoms (sharing a key) and connects the condensed cyclic structure to be formed each other.Bridged ring of the present invention Refer to that sharing two non-adjacent carbon atoms each other by two or more cyclic structures connects the caged scaffold formed.This The invention loop coil refers to that sharing a carbon atom each other by two or more cyclic structures connects the spiral shell formed Ring structure.

" 6~14 yuan of condensed hetero ring bases " of the present invention, refers to that two or more cyclic structures share two each other Adjacent atom (sharing a key) connects the condensed ring at least containing heteroatomic 6~14 annular atom to be formed Structure, including for example " 6~11 yuan of condensed hetero ring bases ", " 6~10 yuan of condensed hetero ring bases ", " 7~10 yuan of condensed hetero ring bases ", " 9~10 yuan thick Heterocyclic radical " etc..Instantiation includes but are not limited to：Imidazolidine simultaneously [4,5-c] pyridine radicals, 3,4- dihydroquinazolines base, 1, 2- dihydro-quinoxalines base, benzo [d] [1,3] dioxa cyclopentenyl, 1,3- dihydroisobenzofurans base, 2H- chromogens alkenyl, 2H- chromogen alkene -2- ketone groups, 4H- chromenes base, 4H- chromene -4- ketone groups, Chromanyl, 4H-1,3- benzoxazinyls, 4,6- dihydros - 1H- furans simultaneously [3,4-d] imidazole radicals, 3a, 4,6,6a- tetrahydrochysene -1H- furans simultaneously [3,4-d] imidazole radicals, 4,6- dihydro -1H- thiophene And [3,4-d] imidazole radicals, 4,6- dihydro -1H- pyrrolo-es [3,4-d] imidazole radicals, 4,5,6,7- tetrahydrochysene -1H- benzos [d] imidazole radicals Deng.

" 5~8 unit's heteroaryl " of the present invention, including for example " 5~7 unit's heteroaryl ", " 5~6 unit's heteroaryl ", " 5~ 6 membered nitrogen-containing heteroaryl bases " etc..Instantiation include but are not limited to furyl, thienyl, pyrrole radicals, thiazolyl, isothiazolyl, Thiadiazolyl group, oxazolyl, isoxazolyl, oxadiazolyls, imidazole radicals, pyrazolyl, 1,2,3- triazolyls, 1,2,4- triazolyls, 1, 2,3- oxadiazolyls, 1,2,4- oxadiazolyls, 1,2,5- oxadiazolyls, 1,3,4- oxadiazolyls, pyridine radicals, 2- pyridones, 4- Pyridone, pyrimidine radicals, 1,4- Dioxins base, 2H-1,2- oxazinyls, 4H-1,2- oxazinyls, 6H-1,2- oxazinyls, 4H-1,3- oxazinyls, 6H-1,3- oxazinyls, 4H-1,4- oxazinyls, pyridazinyl, pyrazinyl, 1,2,3- triazine radicals, 1,3,5- tri- Piperazine base, 1,2,4,5- tetrazine bases, azepine cycloheptatriene base, 1,3- diazas cycloheptatriene base, azepine cyclooctatetraenyl etc., preferably For " 5~6 membered nitrogen-containing heteroaryl base ".

" 6~14 yuan of thick heteroaryls " of the present invention, refers to that two or more cyclic structures share two each other Adjacent atom (sharing a key) connect to be formed at least containing heteroatomic 6~14 annular atoms, tool There are the condensed cyclic structure of armaticity, including such as " 6~10 yuan of thick heteroaryls ", " 7~10 yuan of thick heteroaryls ", " 9~10 yuan of thick heteroaryls Base ", " 9~10 yuan of nitrogenous thick heteroaryls " etc..Instantiation includes but not limited to：Benzofuranyl, benzisoxa furyl, benzo Thienyl, indyl, iso-indoles, benzoxazolyl, benzimidazolyl, indazolyl, benzotriazole base, quinolyl, 2- quinolinones, 4- quinolinones, 1- isoquinolines, isoquinolyl, acridinyl, phenanthridinyl, benzo pyridazinyl, phthalazinyl, quinazolyl, quinoxaline Base, phenol piperazine base, pteridine radicals, purine radicals, naphthyridines base, azophenlyene, phenthazine etc..

" 6~12 yuan of bridge heterocyclic radicals " of the present invention, refers to that any two ring shares two non-conterminous atoms and formed The caged scaffold at least containing heteroatomic 6~12 annular atom, refer to any two ring share two it is non-conterminous Atom formed the cyclic structure at least containing heteroatomic 6~12 annular atom, the hetero atom be selected from N, S, O, CO, SO and/or SO₂Deng.Including such as " 6~11 yuan of bridge heterocyclic radicals ", " 6~9 yuan of bridge heterocyclic radicals ", " 7~10 yuan of bridge heterocycles Base ", " 7~9 yuan of bridge heterocyclic radicals ", " 7~8 yuan of bridge heterocyclic radicals ", " 8 yuan of bridge heterocyclic radicals ", " 8 yuan of nitrogenous bridge heterocyclic radicals " etc..In fact Example includes but not limited to for example： Deng.

" 6~12 yuan of spiro heterocyclic radicals " of the present invention, refers to that at least two rings are shared an atom and formed at least Cyclic structure containing heteroatomic 6~12 annular atom, the hetero atom are selected from N, S, O, CO, SO and/or SO₂ Deng.Including such as " 6~11 yuan of spiro heterocyclic radicals ", " 7~11 yuan of spiro heterocyclic radicals ", " 7~10 yuan of spiro heterocyclic radicals ", " 7~9 yuan Spiro heterocyclic radical ", " 7~8 yuan of spiro heterocyclic radicals ", " 7~8 yuan of nitrogenous spiro heterocyclic radicals " etc..The example includes but are not limited to for example： Deng.

Present invention also offers the preparation method of formula (I) compound, (wherein, it includes but not limited to following process routes It is defined as follows representated by each abbreviation：DCM：Dichloromethane；DIPEA：N, N- diisopropylethylamine；DMSO：Dimethyl sulfoxide (DMSO)； EA：Ethyl acetate；HATU：2- (7- azos benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid esters；MeOH：Methanol； NBS：N- bromo-succinimides；PE：Petroleum ether；THF：Tetrahydrofuran；Xant-phos：Double diphenylphosphine -9,9- the diformazans of 4,5- Base xanthene；x-phos：2- dicyclohexyl phosphorus -2', 4', 6'- tri isopropyl biphenyl)：

Process route：

R¹、R²、R³、R⁴、n、A¹、A²、A³As it was noted above, X represents halogen, selected from fluorine, chlorine, bromine, iodine.

Specific illustrative steps are as follows：

1st, the preparation of intermediate 1

Raw material 1 and organic base are dissolved in organic solvent, raw material 2 is slowly added dropwise under cryogenic conditions, is stirred after reaction, Extraction, organic layer drying, obtains intermediate 1 after concentration, the wherein preferred DCM of organic solvent or Isosorbide-5-Nitrae-dioxane, organic base is preferred For triethylamine.

2nd, the preparation of intermediate 2

Intermediate 1, raw material 3 and organic base are dissolved in organic solvent, phosphorus oxychloride is added dropwise, after reaction, adds alkali, Adjusting pH value, extraction, separates organic phase drying, is concentrated to give intermediate 2, the wherein preferred dichloromethane of organic solvent or 1,2- for neutrality Dichloroethanes, organic base select triethylamine.

3rd, the preparation of intermediate 3

Intermediate 2 is dissolved in organic solvent, add potassium tert-butoxide, be warming up to 100 DEG C reaction 2 it is small when, after reaction, Add water quenching to go out, extract, dry, residue column chromatography obtains intermediate 3, the wherein preferred DCM of organic solvent after concentration.

4th, the preparation of intermediate 4

Intermediate 3 and pinacol borate are dissolved in organic solvent, palladium, thricyclohexyl phosphorus and potassium acetate, nitrogen Under protection, heating reaction.After reaction, water and organic solvent extraction, organic layer drying, concentration, through column chromatography for separation are added Obtain intermediate 4, the wherein preferred DMF of organic solvent or Isosorbide-5-Nitrae-dioxane.

5th, the preparation of intermediate 5

Raw material 4 and intermediate 4 are dissolved in organic solvent, inorganic base and tetrakis triphenylphosphine palladium are added, under nitrogen protection It is heated to reaction to terminate, adds water, extract, dry, concentration, intermediate 5, the wherein preferred Isosorbide-5-Nitrae of organic solvent is obtained through column chromatography for separation Dioxane or DMF.

6th, the preparation of intermediate 6

Raw material 5 is dissolved in organic solvent, benzoyl peroxide is added and NBS is heated to reaction and terminates, filter, concentration, Residue obtains intermediate 6, the wherein preferred carbon tetrachloride of organic solvent through column chromatography for separation.

7th, the preparation of intermediate 7

Intermediate 6 and raw material 6 are dissolved in organic solvent, inorganic base room temperature reaction is added, terminates to reaction, filter, filter Liquid concentrates, and residue column chromatography for separation obtains intermediate 7, the wherein preferred acetonitrile of organic solvent.

8th, the preparation of formula (I) compound

Intermediate 5 and intermediate 7 are dissolved in organic solvent, add three (dibenzalacetone) two palladiums, 2- dicyclohexyls Phosphorus -2', 4', 6'- tri isopropyl biphenyl, cesium carbonate, the lower heating reaction of nitrogen protection.After reaction, water is added, is extracted, is done Dry residue column chromatography for separation obtains formula (I) compound, the wherein preferred Isosorbide-5-Nitrae dioxane of organic solvent or DMF.

" pharmaceutically acceptable salt " of compound shown in formula (I) of the present invention refers to acid present in formula (I) compound The salt that functional group is formed with appropriate inorganic or organic cation (alkali), including formed with alkali or alkaline earth metal salt, Ammonium salt, and the salt formed with nitrogenous organic base；And basic functionality present in formula (I) compound (such as-NH₂Deng) with The salt of appropriate inorganic or organic anion (acid) formation, including with inorganic acid and organic carboxyl acid.

" ester " of compound shown in formula (I) of the present invention refer to, when formula (I) compound is there are during carboxyl, can occur with alcohol Esterification and the ester formed, when formula (I) compound is there are during hydroxyl, can occur with organic acid, inorganic acid, acylate etc. Esterification and the ester formed.Under the conditions of existing for acid or alkali the corresponding acid of hydrolysis generation or alcohol can occur for ester.

" solvate " of compound shown in formula (I) of the present invention refers to its material to be formed that associates with solvent molecule.It is described Solvent can be organic solvent (such as methanol, ethanol, propyl alcohol, acetonitrile etc.), water etc..Such as formula (I) compound of the present invention can be with Alcoholate is formed with ethanol, hydrate is formed with water.

" alloisomerism " of the compounds of this invention is divided into conformation and configuration isomery, and configuration isomery be also divided into cis-trans isomerism and Optical isomerism.Conformational isomerism refers to have the organic molecule of certain configuration to cause due to carbon, the rotation of carbon single bond or distortion Each atom of molecule or atomic group produce a kind of stereo-isomerism of different arrangement modes in space, common are alkane and ring The structure of alkane derivative, such as the chair conformation and boat conformation that occur in cyclohexane structure." stereoisomer ", refers to when this Invention compound contains one or more asymmetric centers, thus can be used as racemic modification and racemic mixture, single mapping Isomers, non-enantiomer mixture and single diastereoisomer.The compounds of this invention has asymmetric center, this kind of not right Title center respectively will independently produce two optical isomers, and the scope of the present invention includes all possible optical isomer and non- Enantiomeric mixture and pure or partial-purified compound.If compound of the present invention contains olefinic double bonds, unless Special instruction, the present invention include cis-isomer and transisomer.Compound of the present invention can be with dynamic isomer Form exists, it is by one or more double-bond shifts and the tie point with different hydrogen.For example, ketone and its Enol forms It is ketone-enol tautomers.Each dynamic isomer and its mixture are included in the compound of the present invention.All formulas (I) The enantiomter of compound, diastereoisomer, racemization isomers, cis-trans-isomer, dynamic isomer, geometric isomer, Epimer and its mixture, are included in the scope of the invention.

Further requirement of the present invention protection include compound shown in formula (I), its pharmaceutically acceptable salt, its ester, its The pharmaceutical composition of solvate and their stereoisomer and one or more pharmaceutical carriers and/or diluent, can be with Pharmaceutically acceptable any formulation is made.Being applied in a manner of oral, parenteral, rectum or transpulmonary administration etc. needs this control The patient for the treatment of.During for being administered orally, conventional solid pharmaceutical preparation, such as tablet, capsule, pill, granule can be made into；Also may be used Oral liquid, such as oral solution, oral suspensions, syrup is made.When oral formulations are made, it is suitable to add Filler, adhesive, disintegrant, lubricant etc..During for parenteral administration, injection, including parenteral solution, injection can be made into With aseptic powdery and concentrated solution for injection.When injection is made, the conventional method production in existing pharmaceutical field can be used, is prepared During injection, additives can be added without, suitable additives can be also added according to the property of medicine.During for rectally, It can be made into suppository etc..During for transpulmonary administration, inhalant or spray etc. can be made into.

Further requirement of the present invention protection include compound shown in formula (I), its pharmaceutically acceptable salt, its ester, its Solvate and their stereoisomer and other one or more antitumor agents and the pharmaceutical composition of immunodepressant. The antitumor agent and immunodepressant, include but not limited to methotrexate (MTX), capecitabine, gemcitabine, doxifluridine, Pemetrexed disodium, pazopanib, Imatinib, Erlotinib, Lapatinib, Gefitinib, Vande Thani, Trastuzumab, shellfish Cut down monoclonal antibody, Rituximab, Herceptin, taxol, vinorelbine, docetaxel, Doxorubicin, hydroxycamptothecin, silk It is rimocidin, epirubicin, pirarubicin, bleomycin, Letrozole, tamoxifen, fulvestrant, music score of Chinese operas Rayleigh, Flutamide, bright Third Rayleigh, Anastrozole, ifosfamide, busulfan, endoxan, Carmustine, Nimustine, Semustine, mustargen, horse Flange, chlorambucil, carboplatin, cis-platinum, oxaliplatin, network platinum, Topotecan, camptothecine, topotecan, everolimus, Xi Luomo This, special cancer fits, Ismipur, 6- thioguanines, imuran, rhzomorph D, daunorubicin, adriamycin, mitoxantrone, wins honour for Mycin, plicamycin or aminoglutethimide.

Present invention also offers the compound shown in formula (I) of the present invention, its pharmaceutically acceptable salt, its ester, its solvent Compound and their stereoisomer are preparing the medicine for the treatment of and/or prevention by cancer-related diseases kinase mediated CDK In application.The relevant disease of cancer is selected from brain tumor, lung cancer, dermoid cancer, carcinoma of urinary bladder, stomach cancer, oophoroma, abdomen Film cancer, cancer of pancreas, breast cancer, head and neck cancer, cervix cancer, carcinoma of endometrium, the carcinoma of the rectum, liver cancer, kidney, adenocarcinoma of esophagus, oesophagus Squamous cell carcinoma, prostate cancer, female reproductive tract cancer, carcinoma in situ, lymthoma, neurofibroma, thyroid cancer, osteocarcinoma, skin Cancer, colon cancer, carcinoma of testis, gastrointestinal stromal tumor, tumor of prostate, mast cell tumor, Huppert's disease, melanoma, Glioma or sarcoma.

The compounds of this invention has the following advantages：

(1) formula (I) compound of the present invention, its pharmaceutically acceptable salt, its ester, its solvate and their solid Isomers has excellent CDK kinase inhibiting activities；

(2) formula (I) compound of the present invention, its pharmaceutically acceptable salt, its ester, its solvate and their solid Isomers shows good biological stability, has good pharmacokinetic property, acts on more longlasting, bioavilability It is high.

(3) formula (I) compound of the present invention, its pharmaceutically acceptable salt, its ester, its solvate or theirs is three-dimensional different Structure body shows good blood-brain barrier passability, and possibility is provided for treatment of the CDK inhibitor as the cancer of the brain.

(4) formula (I) compound of the present invention, its pharmaceutically acceptable salt, its ester, its solvate or theirs is three-dimensional different Structure body shows relatively low toxicity, and drug resistance is good, safe.

Below by way of external zymetology and cytology inhibitory activity experiment the present invention is further explained compound advantageous effect, but This should not be interpreted as to the compounds of this invention only has following beneficial effect.

The external zymetology activity experiment of 1 the compounds of this invention of experimental example

Tester：Part of compounds 1,3 of the present invention, its chemical name and preparation method are shown in that the preparation of each compound is implemented Example.

Comparison medicine：LY2835219, structural formula are shown in background section, are purchased from Wuhan Yong Can bio tech ltd.Under The implication stated representated by the abbreviation in experiment is as follows：

Experimental method：The inhibitory activity that CDK kinases is carried out using Caliper Mobility Shift methods is measured

1.1 times of kinase buffer liquids are prepared：

1) preparation of 1 times of CDK4/9 kinase buffer liquid

The Triton X-100 that HEPES, the mother liquid concentration of the pH 7.5 that mother liquid concentration is 1000mM are 10% are taken respectively, are added Enter ultra-pure water mixing, make the final concentration of 20mM of HEPES, final concentration of the 0.01% of Triton X-100.

2. the preparation of terminate liquid

The coating buffer Coating Reagent#3 (12- used in Caliper instruments that mother liquid concentration is 4% are taken respectively Carry coating buffer in sipper chip), the EDTA that mother liquid concentration is the HEPES of 1000mM pH 7.5, mother liquid concentration is 1M, mother Liquid concentration is 30% Brij-35, adds ultra-pure water and mixes, and makes final concentration of 0.2%, the HEPES of Coating Reagent#3 whole Concentration is 100mM, EDTA final concentration of 50mM, Brij-35 final concentration of 0.015%.The wherein CDK1 of compound 3,2,7, make EDTA mother liquid concentrations are 0.5M, and other conditions are constant.

3.2.5 the preparation of times kinase solution

1) preparation of 2.5 times of CDK4/D3 kinase solutions

The DTT that CDK4/D3 enzyme solutions, mother liquid concentration that mother liquid concentration is 6.421 μM are 1M is taken respectively, adds 1 times of CDK4 Kinase buffer liquid mixes, and makes the final concentration of 5mM of CDK4/D3 enzymes final concentration of 25nM, DTT.

2) preparation of 2.5 times of CDK9/T1 kinase solutions

The DTT that CDK9/T1 enzyme solutions, mother liquid concentration that mother liquid concentration is 7.096 μM are 1M is taken respectively, adds 1 times of CDK9 Kinase buffer liquid mixes, and makes the final concentration of 5mM of CDK9/T1 enzymes final concentration of 37.5nM, DTT.

4.2.5 the preparation of times polypeptide solution

1) preparation of 2.5 times of CDK4/D3 polypeptide solutions

The MgCl that ATP solution, mother liquid concentration that mother liquid concentration is 100mM are 1M is taken respectively₂, mother liquid concentration is 300 μM The polypeptide 8 of FAM marks, adds 1 times of CDK4 kinase buffer liquid and mixes, make final concentration of 552.5 μM of ATP, MgCl₂It is final concentration of 25mM, 8 final concentration of 7.5 μM of polypeptide.

3) preparation of 2.5 times of CDK9/T1 polypeptide solutions

The MgCl that ATP solution, mother liquid concentration that mother liquid concentration is 10mM are 1M is taken respectively₂, mother liquid concentration is 300 μM CTD3 polypeptides, add 1 times of CDK9 kinase buffer liquid and mix, make final concentration of 57.5 μM of ATP, MgCl₂Final concentration of 25mM, CTD3 Final concentration of 7.5 μM of polypeptide.

5.5 times of tester solution are prepared：

The DMSO storing solutions of 10mM testers are taken, the solution that concentration is 50 μM are made with DMSO dilutions, as mother liquor.With DMSO dilutes above-mentioned mother liquor four step by step again, and then each concentration dilutes 10 times with 1 times of kinase buffer liquid respectively, and 5 times of changes are made Polymer solution.Compound 1 is the DMSO storing solutions with 5mM testers, and other conditions are constant.

The reaction of the zymetology of 6.CDK/4//9：

1) it is prepared that the prepared 5 times of tester solution of 5 μ L, 10 μ L are separately added into 384 orifice plates in corresponding hole 2.5 times of kinase solutions, are incubated at room temperature 10 minutes.

2) corresponding Kong Zhongzai is separately added into the prepared 2.5 times of polypeptide solutions of 10 μ L, makes tester final concentration of 1000nM、250nM、63nM、16nM、4nM、1nM、0.2nM、0.1nM、0.02nM、0.004nM.28 DEG C are incubated startup enzyme reaction, When CDK4 reactions 5 are small, CDK9 reacts 90 minutes.

7. zymetology detects：

25 μ L terminate liquids are separately added into each corresponding hole, terminate reaction.

8.Caliper instruments read data, and calculate inhibiting rate by data：

Inhibiting rate=(maximum-sample value)/(maximum-minimum value) × 100, curve plan is carried out using XLfit softwares Close, draw IC₅₀Value.

Maximum：It is not added with the positive control of tester, minimum value：Not enzyme negative control.

Experimental result and conclusion：

The external zymetology inhibitory activity of 1 the compounds of this invention of table

The external zymetology inhibitory activity of 2 the compounds of this invention of table

From table 1,2, compound 1 and compound 3 show CDK kinases inhibitory activity good enough, particularly pair Comparison medicine activity is significantly better than in the activity of CDK9/T1.

The cell in vitro inhibitory activity of 2 the compounds of this invention of experimental example

Comparison medicine：LY2835219, structural formula are shown in background section, are purchased from Wuhan Yong Can bio tech ltd.

Implication representated by the abbreviation of following experiments is as follows：

Experimental method：Using BrdU methods (BrdU cell proliferation test kits, Cell Signaling Technology companies) carry out cell Proliferation detection

1. reagent and compound are prepared

1) 1 times of washing lotion is prepared：

The washing lotion that mother liquid concentration is 20 times is diluted to 1 times of washing lotion with ultra-pure water.

2) 1 times of detection antibody-solutions is prepared：

The BrdU that mother liquid concentration is 100 times is detected into antibody and is diluted to 1 times of detection antibody-solutions with detection antibody diluent.

3) two corresponding anti-solution of 1 times of HRP mark is prepared：

By mother liquid concentration be 100 times of anti-mouse IgG, HRP labelled antibody with HRP labelled antibodies diluted into 1 times The two corresponding anti-solution of HRP marks.

4) 10 times of BrdU solution：

The BrdU solution that mother liquid concentration is 1000 times is diluted to 10 times of BrdU solution with the corresponding culture medium of cell.

5) test compound is prepared：

Prepare test compound stock solutions：The mother liquor of 10mM is configured to 100%DMSO.

Prepare test compound gradient dilution solution：Take 4 times of continuous gradients of test Compound Stock solution DMSO of 10mM dilute Release, concentration is respectively 2.5mM, 625 μM, 156 μM, 39 μM, 9.8 μM, 2.5 μM.The diluted compounds of DMSO of 2 μ L are taken to add respectively 10 times of testers are configured into 198 nutrient solutions of the μ L containing 10%FBS, tester maximum concentration is 100 μM, and DMSO concentration is 1%, totally 7 concentration gradients.

6) culture medium is prepared：

MDA-MB-435S culture mediums：L-15+10%FBS+0.01mg/mL insulin

MCF-7 culture mediums：DMEM+10%FBS+0.01mg/mL insulin

U87MG culture mediums：MEM+10%FBS

Colo205, K562 culture medium：RPMI-1640+10%FBS

MDA-MB-468 culture mediums：L-15+10%FBS

2. test procedure

1) pancreatin digestion grows to 80% cell (exponential phase), and cell is collected by centrifugation.With the culture medium without FBS Cell is resuspended, counts and adjusts 96 orifice plates of inoculation, the μ L of the 4000/hole of U87MG cell inoculations of compound 1/81, compound 3 The μ L of 4000/hole of MDA-MB-468 cell inoculations/81, other are inoculated with the μ L of 3000/hole/81, are placed in 37 DEG C of cell incubators Middle culture；

2) 24 it is small when after per hole add 9 μ L FBS, make FBS final concentration of 10%；

3) 10 times of testers of 10 μ L various concentrations are added per hole, make tester final concentration be respectively 10 μM, 2.5 μM, 625nM, 156nM, 39nM, 9.8nM, 2.5nM, 3 multiple holes/group, when 37 DEG C of cultures 72 are small；

Solvent control：0.1%DMSO

Blank control：Only add culture medium, without cell

Normal cell controls：There is no the normal cell of any processing

4) 10 μ L, 10 times of BrdU solution are added per hole, be incubated in cell incubator 4 it is small when after discard culture medium；K562 is thin Born of the same parents 1000rpm centrifugations discard culture medium after five minutes；

5) 100 μ L are added per hole and fixes/denaturing liquid, be incubated at room temperature 30 minutes, discard solution；

6) 100 μ L, 1 times of detection antibody-solutions are added per hole, when incubation at room temperature 1 is small, solution are discarded, with 1 times of 200 μ of washing lotion L/ is washed in hole 3 times；

7) two corresponding anti-solution of 100 μ L, 1 times of HRP mark is added per hole, is incubated at room temperature 30 minutes, is discarded solution, washed with 1 times 200 μ L/ holes of liquid are washed 3 times；

8) 100 μ L tmb substrate solution are added per hole, are incubated at room temperature 30 minutes；

9) 100 μ L terminate liquids, microplate reader 450nm detection OD values are added per hole.

3. data processing

1) cell survival rate (%)=(OD_Tester-OD_{Blank control})/(OD_{Normal cell controls}-OD_{Blank control}) × 100%, OD_{Blank control}：Blank Control value, OD_{Normal cell controls}：Normal cell controls value；

2) data are mapped using 5 softwares of GraphPad Prism, obtain curve and IC₅₀Value.

Experimental result:

The cell in vitro activity of 3 the compounds of this invention of table

The cell in vitro activity of 4 the compounds of this invention of table

From table 3,4, compound 1 and 3 pairs of cancer cells have good inhibiting effect；The cell in vitro of compound 1 is lived Property is suitable with comparison medicine；The cell in vitro activity of compound 3 is significantly better than comparison medicine.

Embodiment

The embodiment of form by the following examples, makees further specifically the above of the present invention It is bright.But the scope that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to following embodiments.It is all to be based on the above of the present invention The technology realized belongs to the scope of the present invention.

Preparation example 1：The preparation of the bromo- 3- fluorine pyridine -2- amine of 5-

3- fluorine pyridine -2- amine (10.0g, 89.3mmol) is dissolved in acetic acid (100mL), under the conditions of ice-water bath, by liquid Bromine (14.3g, 89.3mmol) is added dropwise in solution.2h is reacted at room temperature, and TLC detection reactions are completed, and add ethyl acetate (80mL), is concentrated under reduced pressure into the 1/3 of original volume, filters, and weighs (12.0g, yield 70.6%) after obtained solid drying.

Preparation example 2：The preparation of the fluoro- 2- nitropyridines of the bromo- 3- of 5-

The concentrated sulfuric acid (50mL) and hydrogen peroxide (50mL) are mixed, the bromo- 3- fluorine pyridine -2- amine (10.0g, 52.36mmol) of 5- It is added dropwise to after being dissolved in the concentrated sulfuric acid (20mL) in mixed solution, reacts 24h, LC-MS detection reactions at room temperature.Reaction solution is poured into broken In ice, with sodium carbonate tune pH=7-8, extracted with ethyl acetate (3 × 200mL), concentration, residue is through silica gel column chromatography (oil Ether:Ethyl acetate=3:1) title compound (5.6g, yield 48.4%) is obtained.

Preparation example 3：The preparation of the bromo- N- isopropyls -2- nitropyridines -3- amine of 5-

The fluoro- 2- nitropyridines (5.6g, 25.34mmol) of the bromo- 3- of 5- are dissolved in n,N-Dimethylformamide (50mL), are added Enter potassium carbonate (7.0g, 50.68mmol), isopropylamine (7.5g, 127.0mmol) is added dropwise, react 3h, LC-MS inspections at room temperature Reaction is surveyed to complete.Reaction solution is poured into water (200mL), is extracted with ethyl acetate (2 × 100mL), organic phase saturation chlorination Sodium washs, anhydrous sodium sulfate drying, concentration, and residue is directly used in react in next step.

Preparation example 4：The bromo- N of 5-³The preparation of-isopropyl -2,3- diamines

The bromo- N- isopropyls -2- nitropyridines -3- amine (6.0g, 23.1mmol) of 5- are dissolved in ethanol (50mL), are added Stannous chloride dihydrate (15.6g, 69.2mmol), is heated to reflux 7h, and LC-MS detection reactions are completed.Solution is concentrated, is added Ethyl acetate (100mL), with sodium bicarbonate solution tune pH=7-8, filtering, liquid separation, concentration, residue is through silica gel column chromatography (stone Oily ether:Ethyl acetate=1:1) title compound (2.5g, yield 47.1%) is obtained.

Preparation example 5：The preparation of bromo- 1- isopropyls -2- methyl isophthalic acids H- imidazos [4,5-b] pyridines of 6-

By the bromo- N of 5-³- isopropyl -2,3- diamines (2.5g, 10.87mmol), acetic acid (0.652g, 10.87mmol) and more Polyphosphoric acid (30mL) mixes, and is heated to 170 DEG C, reacts 2h, and LC-MS detection reactions are completed.Reaction solution is poured into trash ice In (100g), with saturated sodium bicarbonate solution tune pH=7-8, extracted with ethyl acetate (2 × 200mL), organic phase saturation chlorine Change sodium solution washing, concentration, residue is through silica gel column chromatography (petroleum ether:Ethyl acetate=1:2) title compound is obtained (1.5g, yield 54.3%).

Preparation example 6：1- isopropyl -2- methyl -6- (penta ring -2- bases of 4,4,5,5- tetramethyl -1,3,2- dioxies boron) -1H- The preparation of imidazo [4,5-b] pyridine

By bromo- 1- isopropyls -2- methyl isophthalic acids H- imidazos [4, the 5-b] pyridines (200mg, 0.787mmol) of 6-, connection boric acid frequency That alcohol ester (400mg, 1.575mmol) is dissolved in Isosorbide-5-Nitrae-dioxane (20mL), is added [1,1'- double (diphenylphosphine) ferrocene] Palladium chloride (57.6mg, 0.0787mmol) and potassium acetate (231.4mg, 2.361mmol), nitrogen displacement, is heated to 120 DEG C, 12h is reacted, LC-MS detections, which are reacted, to be completed, concentration, adds ethyl acetate (100mL), diatomite filtering, concentrates, residue is direct For reacting in next step.

Preparation example 7：The fluoro- 4- of 5- (1- isopropyl -2- methyl isophthalic acid H- imidazos [4,5-b] pyridine -6- bases) pyridine -2- amine Prepare

By 1- isopropyl-2- methyl-6-(penta ring-2- bases of 4,4,5,5- tetramethyl-1,3,2- dioxies boron)-1H- imidazos [4,5-b] pyridine (593mg, 1.97mmol), the iodo- 5- fluorine pyridines (469mg, 1.97mmol) of 2- amino -4- are dissolved in Isosorbide-5-Nitrae-two In six ring of oxygen (20mL), tetra-triphenylphosphine palladium (25mg, 0.197mmol) and potassium carbonate (815.6mg, 5.91mmol), nitrogen are added Gas is replaced, and is heated to 100 DEG C, when reaction 2 is small, reaction is completed.Concentration, adds ethyl acetate (50mL), filters, and concentration is remaining Thing is through silica gel column chromatography (dichloromethane:Methanol=50:1) title compound (400mg, yield 71.3%) is obtained.

1 N- of embodiment (5- ((4- ethyl piperazidine -1- bases) methyl) pyridine -2- bases) the fluoro- 4- of -5- (1- isopropyl -2- first Base -1H- imidazos [4,5-b] pyridine -6- bases) pyrimidine -2- amine (compound 1) preparation

(1) preparation of 1- [(6- aminopyridine -3- bases) carbonyl] -4- ethyl piperazidines

6- amino-nicotinic acids (13.8g, 0.1mol) and n-ethylpiperazine (11.4g, 0.1mol) are dissolved in DCM (100mL), HATU (57g, 0.15mol) and DIPEA (38.7g, 0.3mol) is added under stirring condition, adds water (20mL) after reacting at room temperature 1h, It is layered to obtain organic phase, silica gel column chromatography (DCM:MeOH=20:1) title compound (18.6g, yield 79.5%) is obtained.

(2) preparation of 1- ((6- aminopyridine -3- bases) methyl) -4- ethyl piperazidines

1- [(6- aminopyridine -3- bases) carbonyl] -4- ethyl piperazidines (18.6g, 0.08mol) are dissolved in tetrahydrofuran (70mL), is cooled to 0 degree, is slowly added to Lithium Aluminium Hydride (6.08g, 0.16mol), and stirring 12h is warmed to room temperature after charging. TLC detection raw material disappear, add water (1mL) reaction is quenched, suction filtration, filtrate decompression it is dry title compound (15g, yield： 85.7%).

(3) preparation of 6- (the chloro- 5-FU -4- bases of 2-) -1- isopropyls -2- methyl isophthalic acid H- imidazos [4,5-b] pyridine

By 1- isopropyl -2- methyl -6- (penta ring -2- bases of 4,4,5,5- tetramethyl -1,3,2- dioxies boron) -1H- imidazos [4,5-b] pyridine (236.9mg, 0.787mmol), 2,4-, bis- chloro- 5-FUs (131.4mg, 0.787mmol) are dissolved in ethylene glycol In dimethyl ether (20mL), two triphenylphosphine palladiums (27.6mg, 0.039mmol) and sodium carbonate liquor (1.18mL are added 2M), nitrogen displacement, is heated to 80 DEG C, reacts 12h, and LC-MS detection reactions are completed.Concentration, adds 50mL ethyl acetate, filters, Concentration, residue is through silica gel column chromatography (dichloromethane:Methanol=100:1) title compound (150mg, yield are obtained 62.3%).

(4) N- (5- ((4- ethyl piperazidine -1- bases) methyl) pyridine -2- bases) the fluoro- 4- of -5- (1- isopropyl -2- methyl isophthalic acids H- Imidazo [4,5-b] pyridine -6- bases) pyrimidine -2- amine preparation

By 6- (the chloro- 5-FU -4- bases of 2-) -1- isopropyls -2- methyl isophthalic acid H- imidazos [4,5-b] pyridine (150.0mg, 0.49mmol), 5- ((4- ethyl piperazidine -1- bases) methyl) pyridine -2- amine (107.8mg, 0.49mmol) are dissolved in 1, In 4- dioxane (20mL), three (dibenzalacetone) two palladiums (22.4mg, 0.0245mmol) of addition, 2- dicyclohexylphosphontetrafluoroborates- 2,4,6- tri isopropyl biphenyls (23.4mg, 0.049mmol) and cesium carbonate (318.5mg, 0.98mmol), nitrogen displacement, heating To 120 DEG C, 12h is reacted, LC-MS detection reactions are completed, and concentration, adds ethyl acetate (100mL), diatomite filtering, concentrates, remain Excess is through silica gel column chromatography (dichloromethane:Methanol=10:1) title compound (40mg, yield 16.7%) is obtained.

Molecular formula：C₂₆H₃₂FN₉Molecular weight：489.6LC-MS(m/z)：490.3(M+H⁺)

¹H-NMR(400MHz,DMSO-d₆)10.13(s,1H),9.00(s,1H),8.72(s,1H),8.71(s,1H), 8.22(s,1H),8.19(s,1H),7.65-7.68(m,1H),4.83-4.87(m,1H),3.39(s,2H),2.67(s,3H), 2.28-2.49 (m, 10H), 1.61 (d, J=6.8Hz, 6H), 0.92-0.98 (m, 3H)

2 5- of embodiment ((4- ethyl piperazidine -1- bases) methyl) the fluoro- 4- of-N-5- (1- isopropyl -2- methyl isophthalic acid H- imidazos [4,5-b] pyridine -6- bases) pyridine -2- bases) pyrimidine -2- amine (compound 2) preparation

The preparation of (1) 5-(bromomethyl)-2- chlorine pyrimidines

The chloro- 5- methylpyrimidines (5.0g, 38.9mmol) of 2- are dissolved in carbon tetrachloride (40mL), add N- bromo succinyl The benzoyl peroxide (100mg) of imines (6.92g, 38.9mmol) and catalytic amount, heated overnight at reflux, is cooled to room temperature, filtering, Filtrate concentrates, and residue is through column chromatography for separation (petroleum ether:Ethyl acetate=1:1) title compound (3.0g, yield, are obtained 37.5%).(2) preparation of the chloro- 5- of 2- ((4- ethyl piperazidine -1- bases) methyl) pyrimidine

In the tetrahydrofuran solution (15mL) of n-ethylpiperazine (276mg, 2.42mmol), diisopropylethylamine is added (624mg, 4.84mmol) and 5-(bromomethyl)-2- chlorine pyrimidines (500mg, 2.42mmol), the reaction was continued at room temperature 4 it is small when, instead It should complete.Concentration, adds ethyl acetate (50mL) and water (30mL), liquid separation, water phase ethyl acetate (30mL) extraction, merges organic Phase, concentration, residue is through column chromatography for separation (petroleum ether:Ethyl acetate=1:1) title compound (300mg, yield, are obtained 51.6%).

(3) 5- ((4- ethyl piperazidine -1- bases) methyl) the fluoro- 4- of-N-5- (1- isopropyl -2- methyl isophthalic acid H- imidazos [4,5- B] pyridine -6- bases) pyridine -2- bases) and pyrimidine -2- amine preparation

By the fluoro- 4- of 5- (1- isopropyl -2- methyl isophthalic acid H- imidazos [4,5-b] pyridine -6- bases) pyridine -2- amine (100.0mg, 0.351mmol), the chloro- 5- of 2- ((4- ethyl piperazidine -1- bases) methyl) pyrimidine (84.2mg, 0.351mmol) are dissolved in In Isosorbide-5-Nitrae-dioxane (15mL), three (dibenzalacetone) two palladiums (32mg, 0.035mmol) of addition, 2- dicyclohexylphosphontetrafluoroborate -2, 4,6- tri isopropyl biphenyls (33.5mg, 0.07mmol) and cesium carbonate (342mg, 1.05mmol), nitrogen displacement, is heated to 110 DEG C, overnight, LC-MS detection reactions are completed, and by reacting liquid filtering, concentration, residue is through silica gel column chromatography (dichloromethane for reaction: Methanol=15:1) title compound (31mg, yield 18.2%) is obtained.

Molecular formula：C₂₆H₃₂FN₉Molecular weight：489.6 LC-MS(m/z)：490.4(M+H⁺)

¹H-NMR(400MHz,CDCl₃)8.80 (d, J=1.6Hz, 1H), 8.71 (d, J=6.0Hz, 1H), 8.44 (s, 2H), 8.27 (d, J=2.4Hz, 1H), 8.09-8.11 (m, 2H), 4.71-4.78 (m, 1H), 3.50 (s, 2H), 2.74 (s, 3H), 2.42-2.70 (m, 9H), 1.69 (d, J=6.8Hz, 6H), 1.09 (t, J=7.2Hz, 3H)

3 N- of embodiment (5- ((4- ethyl piperazidine -1- bases) methyl) pyridine -2- bases) the fluoro- 4- of -5- (1- isopropyl -2- first Base -1H- imidazos [4,5-b] pyridine -6- bases) pyridine -2- amine (compound 3) preparation

The preparation of (1) 5-(bromomethyl)-2- chloropyridines

Chloro--methylpyridine (3.0g, 23.51mmol) is dissolved in carbon tetrachloride (15mL), adds N- bromos fourth two The benzoyl peroxide (100mg) of acid imide (4.185g, 23.51mmol) and catalytic amount, heated overnight at reflux, is cooled to room temperature, Filtering, filtrate concentration, residue are directly used in next step.

(2) preparation of 1- ((6- chloropyridine -3- bases) methyl) -4- ethyl piperazidines

In the tetrahydrofuran solution (50mL) of n-ethylpiperazine (2.685g, 23.51mmol), diisopropylethylamine is added (6.065g, 47.02mmol) and 5-(bromomethyl)-2- chloropyridines (4.854g, 23.51mmol), it is 4 small that the reaction was continued at room temperature When, reaction is completed.Concentration, adds ethyl acetate (50mL) and water (30mL), liquid separation, water phase ethyl acetate (30mL) extraction, is closed And organic phase, concentration, residue is through column chromatography for separation (petroleum ether：Ethyl acetate=1:1) title compound (4.0g, yield, are obtained 71.4%).

(3) N- (5- ((4- ethyl piperazidine -1- bases) methyl) pyridine -2- bases) the fluoro- 4- of -5- (1- isopropyl -2- methyl isophthalic acids H- Imidazo [4,5-b] pyridine -6- bases) pyridine -2- amine preparation

By the fluoro- 4- of 5- (1- isopropyl -2- methyl isophthalic acid H- imidazos [4,5-b] pyridine -6- bases) pyridine -2- amine (100.0mg, 0.351mmol), the dissolving of 1- ((6- chloropyridine -3- bases) methyl) -4- ethyl piperazidines (83.5mg, 0.351mmol) In Isosorbide-5-Nitrae-dioxane (15mL), three (dibenzalacetone) two palladiums (32mg, 0.035mmol) of addition, 2- dicyclohexylphosphontetrafluoroborates- 2,4,6- tri isopropyl biphenyls (33.5mg, 0.07mmol) and cesium carbonate (342mg, 1.05mmol), nitrogen displacement, is heated to 110 DEG C, overnight, LC-MS detection reactions are completed, and by reacting liquid filtering, concentration, residue is through silica gel column chromatography (dichloromethane for reaction Alkane:Methanol=15:1) title compound (28mg, yield 16.4%) is obtained.

Molecular formula：C₂₇H₃₃FN₈Molecular weight：488.6 LC-MS(m/z)：489.4(M+H⁺)

¹H-NMR(400MHz,CDCl₃)8.75(s,1H),8.21(s,1H),8.16(s,1H),8.04-8.07(m, 2H), 7.60 (d, J=8.4Hz, 1H), 7.38 (s, 1H), 7.27-7.29 (m, 1H), 4.70-4.77 (m, 1H), 3.49 (s, 2H), 2.77 (s, 3H), 2.36-2.74 (m, 10H), 1.69 (d, J=6.8Hz, 6H), 1.10-1.16 (m, 3H)

4 N- of embodiment (4- ((4- ethyl piperazidine -1- bases) methyl) phenyl) the fluoro- 4- of -5- (1- isopropyl -2- methyl isophthalic acids Hydrogen-imidazo [4,5-b] pyridine -6- bases) pyrimidine -2- amine (compound 4) preparation

(1) preparation of the fluoro- 4- iodine pyridines of 2,5- bis-

Measure diisopropylamine (17mL, 122mmol) to be added in THF (220mL), be cooled to -20 DEG C, nitrogen protection is lower slow Slow to add n-BuLi (49mL, 122.5mmol), after charging, -20 DEG C of stirring 0.5h, are cooled to -78 DEG C, are slowly added dropwise 4h is stirred at this temperature after the tetrahydrofuran solution (30mL) of 2,5- difluoro pyridines (13.3g, 115mmol).Weigh iodine (32g, THF (100mL) 126mmol) is dissolved in, is slowly dropped at -78 DEG C in above-mentioned reaction solution, 1h is stirred after being added dropwise.Add water It is warmed to room temperature after (10mL) and THF (30mL), adds saturation sodium thiosulfite, be layered to obtain organic phase, water mutually uses ethyl acetate (3 × 100mL) is extracted, and merges organic phase, anhydrous sodium sulfate drying, silica gel column chromatography (PE:EA=50:1) title compound is obtained (13.5g, yield 48%).

(2) preparation of the iodo- 2-aminopyridine of the fluoro- 4- of 5-

The fluoro- 4- iodine pyridines (12.05g, 50mmol) of 2,5- bis- are dissolved in DMSO (40mL), ammonium hydroxide is added under stirring condition (40mL), stirs 12h under the conditions of 90 DEG C of tube sealings.Ethyl acetate (150mL) is added, is layered to obtain organic phase, anhydrous sodium sulfate is dried, Silica gel column chromatography (DCM：MeOH=30:1) title compound (5.95g, yield 50%) is obtained.

(3) preparation of 4- bromobenzyls bromine

4- methyl bromobenzene (2.0g, 11.7mmol) is dissolved in carbon tetrachloride (20mL), adds N- bromo-succinimides (2.08g, 11.7mmol), is heated to 70 DEG C, when reaction 4 is small, concentration, residue is directly used in next step.

(4) preparation of 1- (4- bromophenyls) -4- ethyl piperazidines

By 4- bromobenzyls bromine (2.93g, 11.7mmol), n-ethylpiperazine (3.34g, 29.2mmol) is dissolved in 50mL tetrahydrofurans In, diisopropylethylamine (3.02g, 23.4mmol) is added dropwise under ice-water bath, reaction at room temperature is stayed overnight.Concentration, adds 100mL acetic acid Ethyl ester and 50mL water, liquid separation, concentration, residue column chromatography for separation (petroleum ether:Ethyl acetate=2:1) title compound, is obtained (1.5g, yield 45.28%).

(5) N- (4- ((4- ethyl piperazidine -1- bases) methyl) phenyl) the fluoro- 4- of -5- (1- isopropyl -2- methyl isophthalic acids hydrogen-imidazoles And [4,5-b] pyridine -6- bases) pyrimidine -2- amine preparation

The fluoro- 4- of 5- (1- isopropyls -2- methyl isophthalic acid H- imidazos [4,5-b] pyridine -6- bases) pyridine -2- amine (100.0mg, 0.351mmol), 1- (4- bromophenyls) -4- ethyl piperazidines (99mg, 0.351mmol) are dissolved in Isosorbide-5-Nitrae-dioxane (15mL), Add three (dibenzalacetone) two palladiums (32mg, 0.035mmol), 2- dicyclohexylphosphontetrafluoroborates -2,4,6- tri isopropyl biphenyls (33.5mg, 0.07mmol) and cesium carbonate (342mg, 1.05mmol), nitrogen displacement, are heated to 110 DEG C, reaction is stayed overnight, LC-MS Detection reaction is completed, and by reacting liquid filtering, concentration, residue is through silica gel column chromatography (dichloromethane:Methanol=15:1) marked Inscribe compound (44mg, yield 25.7%).

Molecular formula：C₂₈H₃₄FN₇Molecular weight：487.6 LC-MS(m/z)：488.3(M+H⁺)

¹H-NMR(400MHz,DMSO-d₆)9.15(s,1H),8.51(s,1H),8.31(s,1H),8.23(s,1H), 7.60 (d, J=8.4Hz, 2H), 7.16 (d, J=8.0Hz, 2H), 7.01 (d, J=4.2Hz, 1H), 4.80-4.84 (m, 1H), 3.37 (s, 2H), 2.65 (s, 3H), 2.32-2.49 (m, 10H), 1.58 (d, J=6.8Hz, 6H), 0.92-0.98 (m, 3H)

Claims

1. compound, its pharmaceutically acceptable salt shown in general formula (I), its stereoisomer,

A¹、A²And A³It is independently selected from nitrogen or carbon；

R¹Selected from C_1-4Alkyl；

R²Selected from C_1-4Alkyl；

R⁴Selected from halogen；

R³Selected from optionally by Q²5~6 member heterocyclic ring containing nitrogen bases of substitution, Q²Selected from amino, hydroxyl, trifluoromethyl, cyano group, C_1-4Alkyl Or C_1-4Alkoxy；

N is selected from 0~1 integer.

2. compound as claimed in claim 1, its pharmaceutically acceptable salt, its stereoisomer, wherein

A¹、A²And A³It is independently selected from nitrogen or carbon；

R¹It is isopropyl；

R²Selected from methyl；

R⁴It is fluorine；

N is selected from 0~1 integer.

3. compound as claimed in claim 2, its pharmaceutically acceptable salt, its stereoisomer, wherein

N is 1.

4. compound as claimed in claim 1, its pharmaceutically acceptable salt, its stereoisomer, wherein

R³Selected from optionally by Q²Substituted pyrrolidinyl, piperidyl or piperazinyl, Q²Selected from trifluoromethyl, C_1-4Alkyl or C_1-4Alkane Epoxide；

N is 1.

5. compound as claimed in claim 1, its pharmaceutically acceptable salt, its stereoisomer, the compound are selected from：

6. pharmaceutical composition, comprising the compound described in claim 1-5 any claims, its pharmaceutically acceptable salt, Its stereoisomer and one or more pharmaceutical carriers.

7. pharmaceutical composition as claimed in claim 6, can also contain one or more antitumor agents and immunodepressant, described Antitumor agent and immunodepressant, including methotrexate (MTX), capecitabine, gemcitabine, doxifluridine, pemetrexed two Sodium, pazopanib, Imatinib, Erlotinib, Lapatinib, Gefitinib, Vande Thani, Trastuzumab, bevacizumab, profit are appropriate Former times monoclonal antibody, Herceptin, taxol, vinorelbine, docetaxel, Doxorubicin, hydroxycamptothecin, mitomycin, table are soft Than star, pirarubicin, bleomycin, Letrozole, tamoxifen, fulvestrant, music score of Chinese operas Rayleigh, Flutamide, Leuprorelin, Ah that Bent azoles, ifosfamide, busulfan, endoxan, Carmustine, Nimustine, Semustine, mustargen, melphalan, knurl can Rather, carboplatin, cis-platinum, oxaliplatin, network platinum, Topotecan, camptothecine, topotecan, everolimus, Sirolimus, special cancer fit, Ismipur, 6- thioguanines, imuran, rhzomorph D, daunorubicin, adriamycin, mitoxantrone, bleomycin, general card are mould Element or aminoglutethimide.

8. compound, its pharmaceutically acceptable salt described in claim 1-5 any claims, its stereoisomer are being made Treatment and/or prevention are ready for use on by the application in the medicine of cancer-related diseases kinase mediated CDK, the relevant disease of cancer Disease is selected from brain tumor, lung cancer, dermoid cancer, carcinoma of urinary bladder, stomach cancer, oophoroma, peritoneal cancer, cancer of pancreas, breast cancer, neck Cancer, cervix cancer, carcinoma of endometrium, the carcinoma of the rectum, liver cancer, kidney, adenocarcinoma of esophagus, esophageal squamous cell carcinoma, prostate cancer, female Between genital tract cancer, carcinoma in situ, lymthoma, neurofibroma, thyroid cancer, osteocarcinoma, cutaneum carcinoma, colon cancer, carcinoma of testis, intestines and stomach Matter knurl, tumor of prostate, mast cell tumor, Huppert's disease, melanoma, glioma or sarcoma.