CN106336382B - 4-saturated cyclic substituted aniline protein kinase inhibitor - Google Patents

4-saturated cyclic substituted aniline protein kinase inhibitor Download PDF

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CN106336382B
CN106336382B CN201610541082.XA CN201610541082A CN106336382B CN 106336382 B CN106336382 B CN 106336382B CN 201610541082 A CN201610541082 A CN 201610541082A CN 106336382 B CN106336382 B CN 106336382B
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王永辉
高羽军
周娟
朱研
刘万登
王栋
沈锡明
吴耀东
李春启
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Hangzhou REX Pharmaceutical Co.,Ltd.
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    • C07—ORGANIC CHEMISTRY
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    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • C07—ORGANIC CHEMISTRY
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    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links

Abstract

The invention discloses a compound which can regulate the activity of protein kinase and is used for treating or preventing diseases related to the protein kinase. Specifically, the invention relates to a 4-saturated cyclic group substituted aniline protein kinase inhibitor, belongs to a compound for regulating the activity of Anaplastic Lymphoma Kinase (ALK), and provides a preparation method of the compound and pharmaceutical application of the compound in treating or preventing ALK-related diseases.

Description

4-saturated cyclic substituted aniline protein kinase inhibitor
Technical Field
The present invention relates to compounds that modulate the activity of protein kinases and are useful in the treatment or prevention of diseases associated with protein kinases. Specifically, the invention relates to a 4-saturated cyclic group substituted aniline protein kinase inhibitor, belongs to a compound for regulating the activity of Anaplastic Lymphoma Kinase (ALK), and provides a preparation method of the compound and pharmaceutical application of the compound in treating or preventing ALK-related diseases.
Background
Malignant tumors are a common and frequently occurring disease that seriously threatens human health and are characterized by abnormal proliferation of cells or variant cells. The proliferation, apoptosis, metastasis and the like of tumor cells are closely related to the abnormality of a certain link in a series of signal transduction pathways inside and outside the cells. Among these signaling pathways, an important class of molecules is protein kinases, and abnormalities of protein kinases are closely related to the occurrence, development and prognosis of tumors, and also are the main causes of a series of other human diseases related to inflammation or proliferative responses; the development of drugs targeting protein kinases is a main means for treating related diseases, and many drugs are approved to be on the market, and the drugs have the characteristics of clear target, definite curative effect and high safety, so the drugs are more and more accepted and supported by clinical medical practice.
Anaplastic Lymphoma Kinase (ALK) is an important member of the protein kinase family, and prior studies have shown that overexpression, mutations and fusion proteins of ALK are directly associated with a variety of tumors, including but not limited to neuroblastoma, Anaplastic Large Cell Lymphoma (ALCL), non-small cell lung cancer (NSCLC), and Inflammatory Myofibroblastoma (IMT), among others. First-generation medicines of Crizotinib (Crizotinib) and second-generation medicine of Ceritinib (Ceritinib) aiming at the ALK fusion gene are respectively marketed in 2011 and 2014, and remarkable progression-free survival and objective effective rate are obtained when the Crizotinib and Ceritinib are used for treating ALK positive lung cancer patients, so that the clinical value of the target spot is confirmed. Despite the remarkable drug effect, due to the heterogeneous characteristics of the tumor and the adaptation of tumor cells to the environmental stress, more and more research reports show that the continuous development of tumor resistance and diseases is almost the inevitable fate of patients of the type; in addition, the serious adverse reactions of the existing medicines, such as high incidence rate of adverse reactions in the digestive tract, hepatotoxicity, prolongation of QT interval and the like, also limit the application of the medicines. In view of the above, there are significant social benefits and values in developing new compounds having excellent ALK inhibitory activity and safety and in developing them to the market to cope with the above problems.
Disclosure of Invention
The invention aims to provide a 4-saturated cyclic group substituted aniline protein kinase inhibitor with a novel structure, and a series of compounds with anti-tumor activity are synthesized and screened through group substitution modification.
In order to realize the purpose, the invention adopts the following technical scheme:
a4-saturated cyclic group substituted aniline protein kinase inhibitor is a compound with the following structural general formula I and pharmaceutically acceptable salts thereof:
Figure BDA0001042691910000021
wherein R is1Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, cyano, aryl, heterocyclyl, - (CH)2)wOR10、-(CH2)wNR10R11、-CO2R10、-CONR10R11、-(CH2)wSO2R10、-(CH2)wSO2NR2R10One or more of the above; r2、R3、R4、R5、R6Each independently selected from hydrogen, halogen, C1-6Alkyl, halo C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy radical, C2-6Alkenyl radical, C2-6Alkynyl, cyano or amino; r7Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, C1-6Alkoxy, halo C1-6Alkoxy, -NR2COR8、-NR2CONR2R8、-NR2SO2R8、-COR8、-CONR2R8、-SO2R8、-SO2NR2R8、-POR8R9、
Figure BDA0001042691910000022
R8、R9Each independently selected from hydrogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6One or more of alkynyl, hydroxyl and aryl; r10、R11Each independently selected from hydrogen and C1-6Alkyl, halo C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, aryl or heterocyclyl; heterocyclyl is a 3-12 membered heterocyclic ring selected from the group consisting of the heteroatoms N, O; and n is selected from any integer value of 1-6, and w is selected from any integer value of 0-3.
Preferably, in the above general structural formula I, R1Selected from hydrogen, halogen, C1-6Alkyl, cyano, phenyl, heterocyclyl, -CO2R10、-(CH2)wOR10、-(CH2)wNR10R11、-CONR10R11、-(CH2)wSO2R10Or- (CH)2)wSO2NR2R10；R2、R3、R4、R5、R6Each independently selected from hydrogen, halogen, C1-6Alkyl, halo C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy, cyano or amino; r7Selected from hydrogen, C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy, -SO2R8、-POR8R9、-SO2NR8R9、-CONR8R9、
Figure BDA0001042691910000023
R8、R9Each independently selected from hydrogen, halogen, C1-6One or more of alkyl, hydroxyl and phenyl; r10、R11Each independently selected from hydrogen and C1-6Alkyl, halo C1-6Alkyl, phenyl or heterocyclyl; heterocyclyl is a 3-6 membered heterocyclic ring selected from the group consisting of the heteroatoms N, O; n is an integer value selected from 1 to 6, and w is an integer value selected from 0 to 3The value is obtained.
A4-saturated cyclic group substituted aniline protein kinase inhibitor also comprises a compound with the following structural general formula II and pharmaceutically acceptable salts thereof:
Figure BDA0001042691910000024
wherein R is1Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, cyano, aryl, heterocyclyl, -CO2R10、-(CH2)wOR10、-(CH2)wNR10R11、-CONR10R11、-(CH2)wSO2R10、-(CH2)wSO2NR2R10One or more of the above; r2、R3、R4、R5、R6Each independently selected from hydrogen, halogen, C1-6Alkyl, halo C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy radical, C2-6Alkenyl radical, C2-6Alkynyl, cyano or amino; r7Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, C1-6Alkoxy, halo C1-6Alkoxy, -NR2COR8、-NR2CONR2R8、-NR2SO2R8、-COR8、-CONR2R8、-SO2R8、-SO2NR2R8、-POR8R9、
Figure BDA0001042691910000031
R8、R9Each independently selected from hydrogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6One or more of alkynyl, hydroxyl and aryl; r10、R11Each independently selected from hydrogen and C1-6Alkyl, halo C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, aryl or heterocyclyl; q is-O-, -S-or-NR0-；R0Selected from hydrogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6One or more of alkynyl, acylamino, hydroxyl, aryl and heterocyclic radical; heterocyclyl is a 3-12 membered heterocyclic ring selected from the group consisting of the heteroatoms N, O; m and n are independently selected from any integer value of 0-6, and m and n are not 0 at the same time.
Preferably, in the above general structural formula II, R1Selected from hydrogen, halogen, C1-6Alkyl, cyano, phenyl, heterocyclyl, -CO2R10、-(CH2)wOR10、-(CH2)wNR10R11、-CONR10R11、-(CH2)wSO2R10Or- (CH)2)wSO2NR2R10；R2、R3、R4、R5、R6Each independently selected from hydrogen, halogen, C1-6Alkyl, halo C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy, cyano or amino; r7Selected from hydrogen, C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy, -SO2R8、-POR8R9、-SO2NR8R9、-CONR8R9、
Figure BDA0001042691910000032
R8、R9Each independently selected from hydrogen, halogen, C1-6One or more of alkyl, hydroxyl and phenyl; r10、R11Each independently selected from hydrogen and C1-6Alkyl, halo C1-6Alkyl, phenyl or heterocyclyl; q is-O-, -S-or-NR0-；R0Selected from hydrogen, C1-6One or more of alkyl, acylamino, hydroxyl, phenyl and heterocyclic radical; heterocyclyl is a 3-6 membered heterocyclic ring selected from the group consisting of the heteroatoms N, O; m and n are independently selected from any integer value of 0-6, and m and n are not 0 at the same time.
Preferably, aryl is phenyl, naphthyl or anthracenyl; heterocyclyl is morpholinyl, piperidinyl, piperazinyl, pyrrolidinyl, azetidinyl, pyranyl, furanyl, pyridinyl or pyrimidinyl.
Preferably, the halogen is one or more of fluorine, chlorine, bromine and iodine.
A4-saturated cyclic group substituted aniline protein kinase inhibitor is selected from the following characteristic compounds with the numbers of REX-B1-REX-B28:
REX-B1: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B2: 5-chloro-N2- (4- (1- ((dimethylamino) methyl) cyclopropyl) -2-isopropoxy-5-methylbenzene) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B3: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((4-oxopiperidinyl) methyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B4: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methanolpiperazinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B5: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methylenepiperazinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B6: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methylenepiperidinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B7: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methylenepyrrolidinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B8: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methyleneazetidinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B9: 5-chloro-N2- (2-iso)Propoxy-5-methyl-4- (1- (((4-tetrahydropyranyl) amino) methyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B10: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (morpholinocyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B11: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((4-tetrahydropyranyl) amino) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B12: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (oxo-4-tetrahydropiperidinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B13: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (2- (4-piperidinylcyclopropyl)) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B14: 5-chloro-N2- (2-trifluoromethoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B15: 5-chloro-N2- (2-isopropoxy-5-chloro-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B16: 5-trifluoromethyl-N2- (2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B17: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4- (2- (dimethylphosphite) phenyl) -2, 4-diaminopyrimidine;
REX-B18: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4- (2- (1- (ethoxymethyl) cyclopropyl) phenyl) -2, 4-diaminopyrimidine;
REX-B19: 5-chloro-N2- (2-isopropoxy)5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4- (2- (N-isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B20: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4- (2- (hydroxyethylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B21: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((methanesulfonyl) methyl) cyclopropyl) phenyl) -N4- (2- (N-isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B22: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (ethoxymethyl) cyclopropyl) phenyl) -N4- (2- (N-isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B23: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((trifluoromethoxy) methyl) cyclopropyl) phenyl) -N4- (2- (N-isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B24: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (carboxy) cyclopropyl) phenyl) -N4- (2- (N-isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B25: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (aminomethyl) cyclopropyl) phenyl) -N4- (2- (N-isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B26: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (4-oxotetrahydropyran) methyl) cyclopropyl) phenyl) -N4- (2- (N-isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B27: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((methylamino) methyl) cyclopropyl) phenyl) -N4- (2- (N-isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B28: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (carboxamide) cyclopropyl) phenyl) -N4- (2- (N-isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B29: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((4-oxopyrrolidinyl)) Methyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B30: 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((4-oxohydroxyazetidinyl) methyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
REX-B31: 5-trifluoromethyl-N2- (2-isopropoxy-5-methyl-4- (1- ((4-oxopiperidinyl) methyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine.
The structural formula of the compounds numbered REX-B1-REX-B31 is shown as follows:
Figure BDA0001042691910000051
Figure BDA0001042691910000061
the invention also provides a synthesis method of the compound shown as the general formula I and the general formula II, and the general reaction route is as follows: general formula I:
Figure BDA0001042691910000062
general formula II:
Figure BDA0001042691910000071
based on the general reaction route, the method comprises the following synthetic schemes:
(1) synthesis scheme 1: synthesis of Compounds 1-3
Step 1: will be provided with
Figure BDA0001042691910000072
(i.e., Compound 1-1) and
Figure BDA0001042691910000073
dissolving in organic solvent, slowly adding sodium hydride, reacting at normal temperature for N hours, adding extractant, extracting, drying, decompressing, and spin-drying to obtain the compound 1-2.
Step 2: dissolving the compound 1-2 in an organic solvent, slowly adding a catalyst, replacing nitrogen, introducing hydrogen, reacting at normal temperature for N hours, adding an extracting agent, extracting, drying, decompressing, and spin-drying to obtain the compound 1-3.
In the synthesis scheme 1, the organic solvent is selected from one or more of dichloromethane, N-dimethylformamide, methanol and dioxane; the extractant is selected from one or more of dichloromethane, pure water and ethyl acetate; the catalyst is selected from one or more of palladium carbon, 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene and palladium acetate.
In the synthesis scheme 1, the reaction temperature at normal temperature is 20-30 ℃, and the reaction time N is 3-10 hours.
(2) Synthesis scheme 2: synthesis of Compound 2-2
Step 1: will be provided with
Figure BDA0001042691910000074
(i.e., Compound 2-1) and
Figure BDA0001042691910000075
dissolving in organic solvent, slowly adding sodium hydride and pyrimidine compounds, displacing with nitrogen, heating for N hours, adding a little ice water to destroy excessive sodium hydride, adding extractant, extracting, drying, decompressing, and spin-drying to obtain compound 2-2.
In the synthesis scheme 2, the organic solvent is selected from one or more of dichloromethane, N-dimethylformamide, methanol and dioxane; the extractant is selected from one or more of dichloromethane, pure water and ethyl acetate; the catalyst is selected from one or more of palladium carbon, 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene and palladium acetate.
In the synthesis scheme 2, the heating reaction temperature is 60-120 ℃, and the reaction time N is 12-16 hours.
(3) Synthesis scheme 3: synthesis of target Compound
Step 1: dissolving the compound 1-3 prepared in the synthesis scheme 1 and the compound 2-2 prepared in the synthesis scheme 2 in an organic solvent, continuously adding cesium carbonate and a catalyst, and carrying out nitrogen replacement and microwave heating reaction for N hours; after the reaction is finished, filtering under reduced pressure to remove redundant catalyst, and performing column chromatography to obtain the target compound (i.e. the compound of the general formula I or the general formula II).
In the synthesis scheme 3, the organic solvent is selected from one or more of dichloromethane, N-dimethylformamide, methanol and dioxane; the catalyst is selected from one or more of palladium carbon, 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene and palladium acetate.
In the synthesis scheme 3, the reaction time is 10-20 hours, preferably 16 hours.
In schemes 1,2 and 3, R1Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, cyano, aryl, heterocyclyl, - (CH)2)wOR10、-(CH2)wNR10R11、-CO2R10、-CONR10R11、-(CH2)wSO2R10、-(CH2)wSO2NR2R10One or more of the above; r2、R3、R4、R5、R6Each independently selected from hydrogen, halogen, C1-6Alkyl, halo C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy radical, C2-6Alkenyl radical, C2-6Alkynyl, cyano or amino; r7Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, C1-6Alkoxy, halo C1-6Alkoxy, -NR2COR8、-NR2CONR2R8、-NR2SO2R8、-COR8、-CONR2R8、-SO2R8、-SO2NR2R8、-POR8R9、
Figure BDA0001042691910000081
R8、R9Each independently selected from hydrogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6One or more of alkynyl, hydroxyl and aryl; r10、R11Each independently selected from hydrogen and C1-6Alkyl, halo C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, aryl or heterocyclyl; q is-O-, -S-or-NR0-；R0Selected from hydrogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6One or more of alkynyl, acylamino, hydroxyl, aryl and heterocyclic radical; heterocyclyl is a 3-12 membered heterocyclic ring selected from the group consisting of the heteroatoms N, O; m and n are independently selected from any integer value of 0-6, and m and n are not 0 at the same time.
The term "compound" as used herein includes all stereoisomers, geometric isomers, tautomers and isotopes.
The "compounds" of the present invention may be asymmetric, e.g., having one or more stereoisomers. Unless otherwise indicated, all stereoisomers include, for example, enantiomers and diastereomers. The compounds of the invention containing asymmetric carbon atoms can be isolated in optically active pure form or in racemic form. The optically active pure form can be resolved from a racemic mixture or synthesized by using chiral starting materials or chiral reagents.
The "compound" of the present invention also includes tautomeric forms. Tautomeric forms result from the exchange of one single bond with an adjacent double bond and the concomitant migration of one proton.
The invention also includes all isotopic atoms, whether in the intermediate or final compound. Isotopic atoms include those having the same atomic number but different mass numbers. For example, isotopes of hydrogen include deuterium and tritium.
Compounds containing the foregoing general structure, the terms used herein have the following meanings:
the term "halogen" denotes fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or bromine.
The term "cyano" refers to — CN.
The term "hydroxy" refers to-OH.
The term "alkyl" denotes a straight or branched chain saturated hydrocarbon group consisting of carbon and hydrogen atoms, such as C1-20Alkyl, preferably C1-6Alkyl groups such as methyl, ethyl, propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, sec-butyl or tert-butyl), pentyl (including n-pentyl, isopentyl, neopentyl), n-hexyl, 2-methylhexyl and the like. The alkyl group may be unsubstituted or substituted with one or more substituents including, but not limited to, alkyl, alkoxy, cyano, hydroxy, carbonyl, carboxy, aryl, heteroaryl, amino, halo, sulfonyl, sulfinyl, phosphoryl.
The term "amino" refers to the group-NH2-NH (alkyl) and-N (alkyl)2Alkyl has the meaning as previously described. The structural form of-NH (alkyl) is
Figure BDA0001042691910000091
Specific examples include, but are not limited to-NHCH3、-NHCH(CH3)2、-NHC2H5Etc.; -N (alkyl)2In the structural form of
Figure BDA0001042691910000092
Specific examples include, but are not limited to, -N (CH)3)2、-N(CH3)C2H5And the like.
The term "aryl" refers to an all-carbon monocyclic or fused ring having a fully conjugated pi-electron system, typically having 6 to 14 carbon atoms, preferably having 6 to 12 carbon atoms, and most preferably having 6 carbon atoms. Aryl groups may be unsubstituted or substituted with one or more substituents including, but not limited to, alkyl, alkoxy, cyano, hydroxy, carbonyl, carboxy, aryl, aralkyl, amino, halo, sulfonyl, sulfinyl, phosphoryl. Examples of unsubstituted aryl groups include, but are not limited to, phenyl, naphthyl, and anthracenyl.
The term "heterocyclyl" refers to a monocyclic or fused ring having from 3 to 12 (an integer) ring atoms, of which 1,2 or 3 ring atoms are selected from one or more of N, O, the remaining ring atoms being C, and having a fully conjugated pi-electron system. The heterocyclyl group may be unsubstituted or substituted with one or more substituents including, but not limited to, alkyl, alkoxy, cyano, hydroxy, carbonyl, carboxy, aryl, aralkyl, amino, halo, sulfonyl, sulfinyl, phosphoryl. Examples of unsubstituted heterocyclyl groups include, but are not limited to, pyrrolyl, indolyl, pyrrolidinyl, imidazolyl, pyrazolyl, tetrazolyl, pyridyl, quinolyl, isoquinolyl, piperidyl, pyrimidinyl, pyrazinyl, piperazinyl, furyl, pyranyl, morpholinyl.
The invention also provides a pharmaceutical composition, which comprises the compound or the pharmaceutically acceptable salt thereof as an active ingredient, and one or more pharmaceutically acceptable carriers.
"pharmaceutical composition" as used herein, refers to a formulation of one or more compounds of the present invention or salts thereof with a carrier generally accepted in the art for delivery of biologically active compounds to an organism (e.g., a human). The purpose of the pharmaceutical composition is to facilitate delivery of the drug to an organism.
The term "pharmaceutically acceptable carrier" refers to a substance that is co-administered with, and facilitates the administration of, an active ingredient, including, but not limited to, any glidant, sweetener, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersant, disintegrant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that is acceptable for use in humans or animals (e.g., livestock) as permitted by the national food and drug administration. Examples include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.
The pharmaceutical composition can be prepared into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powder, granules, paste, emulsions, suspensions, solutions, suppositories, injections, inhalants, gels, microspheres, aerosols and the like.
The pharmaceutical compositions of the present invention may be manufactured by methods well known in the art, such as conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, lyophilizing, and the like.
The route of administration of the compounds of the present invention or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof includes, but is not limited to, oral, rectal, transmucosal, enteral, or topical, transdermal, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration. The preferred route of administration is oral.
For oral administration, the pharmaceutical compositions may be formulated by mixing the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, slurries, suspensions and the like, for oral administration to a patient. For example, for pharmaceutical compositions intended for oral administration, tablets may be obtained in the following manner: the active ingredient is combined with one or more solid carriers, the resulting mixture is granulated if necessary, and processed into a mixture or granules, if necessary with the addition of small amounts of excipients, to form tablets or tablet cores. The core may be combined with an optional enteric coating material and processed into a coated dosage form more readily absorbed by an organism (e.g., a human).
The invention also provides the use of a compound as described hereinbefore or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of a disease associated with a protein kinase.
Use of a compound as described above or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of a disease associated with anaplastic lymphoma kinase (ALK kinase).
Preferably, the ALK kinase-associated disease is selected from cell proliferative diseases, preferably tumors.
Preferably, the aforementioned cell proliferative diseases include non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumors, nasopharyngeal carcinoma, breast cancer, colorectal cancer, diffuse large B cell lymphoma, liver cancer, gastric cancer, esophageal cancer, pancreatic cancer, ovarian cancer, systemic histiocytosis and neuroblastoma.
In the invention, the inventor carries out ALK kinase inhibition activity and ALK related mutation site binding rate determination experiments on a series of synthesized 4-saturated cyclic substituted aniline compounds, and finds that part of the compounds show higher ALK inhibition activity, show better binding rate on ALK mutation sites (such as L1196M, F1174L and C1156Y) and have obvious inhibition activity on LTK and ROS 1; in addition, cell proliferation experiments of lung cancer cell strains and phenotype screening experiments of zebra fish are also carried out, and the anti-tumor activity of part of compounds in vivo is found to be obvious.
Compared with the prior art, the 4-saturated cyclic group substituted aniline protein kinase inhibitor provided by the invention is based on reasonable drug design of a target, and a series of compounds with novel structures are obtained through substitution modification of groups; and a series of compounds with antitumor activity are optimally screened out by combining a kinase activity experiment, a cell proliferation experiment and a zebra fish phenotype screening experiment. Therefore, the complex can be used for developing a new generation of protein kinase inhibitor, has great clinical application value for targeted treatment or prevention of ALK-mediated diseases, and has considerable market potential.
Drawings
FIG. 1 is a dorsal distribution of Albino zebrafish iris pigment cells
FIG. 2 is a dose-effect relationship diagram of the effect of REX-B1/REX-B2 on zebra fish iris pigment cells
FIG. 3 is a graph showing the effect of REX-B1/REX-B2 on pigmented cells of iris of zebra fish
Detailed Description
The following are specific examples of the present invention and further describe the technical solutions of the present invention, but the scope of the present invention is not limited to these examples. All changes, modifications and equivalents that do not depart from the spirit of the invention are intended to be included within the scope thereof.
In the preparation method of the target compound, the liquid chromatography adopts a Waters symmetry C18 chromatographic column. Thin layer chromatography was performed using GF254(0.25 mm). Nuclear Magnetic Resonance (NMR) was measured using a Bruker-400 NMR spectrometer; liquid chromatography/Mass Spectrometry (LC/MS) Using a Waters ZQ mass spectrometer (column: Waters symmetry C18, mm, 5 μm, 35 ℃ C.), the ESI (+) ion mode was used.
In addition, all operations involving easily oxidizable or hydrolyzable raw materials were carried out under nitrogen protection. Unless otherwise indicated, the starting materials used in the present invention are all commercially available starting materials and can be used without further purification.
Example 15 chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4Preparation of (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine [ numbered REX-B1 ]
The synthetic route is as follows:
Figure BDA0001042691910000111
synthesis scheme 1: synthesis of intermediate 2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) aniline (i.e. compound 1-12)
Step 1: preparation of intermediate 2-methyl-5-nitro-phenylacetic acid (i.e., Compound 1-2)
Dissolving a raw material 2-methyl-phenylacetic acid (200.0g,1.33mol) in dichloromethane (700mL), slowly adding concentrated sulfuric acid (584mL) below 0 ℃, continuing to react for 0.5 hour after the addition is finished, then dropwise adding concentrated nitric acid (30mL), and keeping the temperature at-2-3 ℃ for reacting for 16 hours; after completion of the reaction, the reaction mixture was poured into 500mL of water, extracted with dichloromethane, dried, and concentrated to obtain compound 1-2(90.0g) in yield: 34.6 percent.
MSm/z[ESI]：196.1[M+1]。
Step 2: preparation of intermediate 2-methyl-5-nitro-phenylacetic acid methyl ester (i.e. compound 1-3)
Dissolving the compound 1-2(20.0g,102mmol) in methanol (250mL), slowly adding thionyl chloride (27.5g,204mmol) at the temperature of 0-5 ℃, and reacting at the temperature of 75-85 ℃ for 3 hours; after the reaction was completed, the temperature was reduced to room temperature, water was added and ethyl acetate was added for extraction, the organic phase was washed twice with sodium hydrogencarbonate, dried over anhydrous sodium sulfate and concentrated to obtain compound 1-3(21.0g), yield: 98.0 percent.
MSm/z[ESI]：201.2[M+1]。
And step 3: preparation of intermediate methyl 1- (2-methyl-5-nitro) cyclopropylphenylacetate (i.e., Compound 1-4)
Dissolving the compounds 1-3(10.0g,47.8mmol) in N, N-dimethylformamide (DMF, 100mL) under ice-water bath condition, slowly adding sodium hydride (3.8g,95.6mmol) into the reaction system, and stirring at 0 ℃ for 15 minutes; 1, 2-dibromoethane (26.0g,143.4mmol) is dripped into the reaction system and heated to 45 ℃ for reaction for 3 hours; after cooling, the mixture was poured into 50mL of water, extracted with ethyl acetate, dried, concentrated, and subjected to silica gel column chromatography to obtain compound 1-4(2.0g), yield: 17.8 percent.
MSm/z[ESI]：236.5[M+1]。1H-NMR(400MHz,CDCl3)：δ＝8.12(s,1H)，8.07-8.05(d,1H,J＝8.4Hz)，7.35-7.33(d,1H,J＝8.4Hz)，3.65(s,3H)，2.44(s,3H)，1.794-1.787(t,2H,J＝2.8Hz)，1.233-1.226(t,2H,J＝2.8Hz)。
And 4, step 4: preparation of intermediate methyl 1- (2-methyl-5-amino) cyclopropylphenylacetate (i.e., Compound 1-5)
Adding compounds 1-4(2.0g,8.5mmol) and 10% palladium on carbon (i.e. Pd/C,0.4g) into dry methanol (50mL), displacing with nitrogen, introducing hydrogen, and reacting at 25 deg.C for 12 hours; after cooling, the mixture was separated by filtration through celite, dried over anhydrous sodium sulfate, and concentrated to obtain compound 1-5(1.5g) in yield: 86.0 percent.
MSm/z[ESI]：206.2[M+1]。
And 5: preparation of intermediate methyl 1- (2-methyl-5-hydroxy) cyclopropylphenylacetate (i.e., Compound 1-6)
Under the condition of ice-water bath, dissolving the compound 1-5(7.1g,34.89mmol) in sulfuric acid (aq, 6.25%, 240mL) and sodium nitrite (2.8g,41.87mmol) are slowly added into the reaction system, and stirring is carried out for 15 minutes at 0 ℃; copper sulfate (1mol/L,150ml) is dripped into the reaction system to react for 2 hours at 65 ℃; after cooling, the mixture was poured into 500mL of water, extracted with ethyl acetate, dried, concentrated, and subjected to silica gel column chromatography to obtain compounds 1 to 6(4.3g) in yield: 57.6 percent.
MSm/z[ESI]：207.2[M+1]。1H-NMR(400MHz,DMSO-d6)：δ＝7.039-7.019(d,1H,J＝8Hz)，6.754-6.747(s,1H)，6.692-6.665(d,1H,J＝10.8Hz)，5.317(s,1H)，3.66(s,3H)，2.252(s,3H)，1.703-1.687(t,2H,J＝3.2Hz)，1.187-1.161(t,2H,J＝3.6Hz)。
Step 6: preparation of intermediate methyl 1- (2-methyl-4-nitro-5-hydroxy) cyclopropylphenylacetate (i.e., Compound 1-7)
Under the ice-water bath condition, the compounds 1 to 7(4.3g,20.85mmol) were dissolved in dichloromethane (100ml), and nitric acid (10.4ml,20.8mmol,2mol/L) was slowly added to the reaction system, stirred at 0 ℃ for 15 minutes, and then reacted at 65 ℃ for 2 hours; after the reaction, the reaction mixture was cooled and poured into 100mL of water, extracted with ethyl acetate, dried, concentrated, and subjected to silica gel column chromatography to obtain compounds 1 to 7: (2.5g), yield: 47.3 percent.
MSm/z[ESI]：252.2[M+1]。
And 7: preparation of intermediate methyl 1- (2-methyl-4-nitro-5-isopropoxy) cyclopropylphenylacetate (i.e., Compound 1-8)
Compound 1-7(2.5g, 9.87mmol) and potassium carbonate (4.1g,29.61mmol) were dissolved in dichloromethane (60ml), isopropyl iodide (5.1g,29.61mmol) was slowly added to the reaction system, stirred at 0 ℃ for 15 minutes, followed by reaction at 25 ℃ for 2 hours; after completion of the reaction, purified water and methylene chloride were added to conduct extraction, followed by drying, concentration and silica gel column chromatography to obtain compounds 1 to 8(2.8g) in yield: 100.0 percent.
MSm/z[ESI]：294.2[M+1]。1H-NMR(400MHz,CDCl3)＝δ7.602(s,1H)，6.969(s,1H)，4.629(m,1H)，3.652(s,3H)，2.287(s,3H)，1.75-1.743(t,2H,J＝2.8Hz)，1.39-1.375(d,6H,J＝6Hz)，1.172-1.165(t,2H,J＝2.8Hz)。
And 8: preparation of intermediate 1- (2-methyl-4-nitro-5-isopropoxy) cyclopropylphenylacetic acid (i.e., compound 1-9)
Compound 1-8(1.0g,3.4mmol) and lithium hydroxide (571.0mg,13.6mmol) were dissolved in a mixture of methanol (50ml) and pure water (10ml), and the reaction was heated to 80 ℃ and stirred for 1 hour; after completion of the reaction, 1mol/L hydrochloric acid was added to adjust pH to 4, and the mixture was filtered to obtain compound 1-9(750.0mg), yield: 79.0 percent.
MSm/z[ESI]：280.2[M+1]。
And step 9: preparation of intermediate (1- (5-isopropoxy-2-methyl-4-nitro) cyclopropyl) phenylacetic acid morpholine (i.e. compound 1-10)
Dissolving the compounds 1-9(600.0mg,2.15mmol), triethylamine (653.0mg,6.45mmol), 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (1.1g,2.8mmol) and morpholine (281.0mg,3.23mmol) in N, N-dimethylformamide (50ml) and reacting at room temperature for 3 hours; after completion of the reaction, extraction was performed with pure water and ethyl acetate, and drying and spin-drying were performed to obtain compounds 1 to 10(673.0mg) in yield: 90.0 percent.
MSm/z[ESI]：347.4[M+1]。
Step 10: preparation of intermediate 2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) nitrobenzene (i.e. compound 1-11)
Dissolving compound 1-10(673.0mg,1.93mmol) in tetrahydrofuran (50ml), and adding borane tetrahydrofuran solution (14.4ml,14.4mmol,1mol/L) dropwise at room temperature; after the reaction was completed, extraction was performed with pure water and ethyl acetate, and drying and spin-drying were performed to obtain compounds 1 to 11(450.0mg), yield: 69.0 percent.
MSm/z[ESI]：333.4[M+1]。
Step 11: preparation of intermediate 2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) aniline (i.e. compound 1-12)
Compounds No. 1 to 11(450.0mg,1.35mmol) and palladium on charcoal (238.0mg) were dissolved in methanol (25ml), and the mixture was replaced with hydrogen overnight at room temperature; after the reaction was completed, the palladium-carbon was removed by filtration, dried and spin-dried to obtain compounds 1 to 12(400.0mg) with yield: 97.0 percent.
MSm/z[ESI]：305.4[M+1]。
Synthesis scheme 2: synthesis of intermediate 2, 5-dichloro-N- [2- [ (1-methylethyl) sulfonyl ] phenyl ] -4-aminopyrimidine (i.e. compound 2-5)
Step 1: preparation of intermediate 2- (isopropylsulfide group) nitrobenzene (i.e. compound 2-2)
Adding raw materials of 2-fluoronitrobenzene (namely a compound 2-1,10.0g,70.0mmol), isopropyl mercaptan (5.4g,70.0mmol) and potassium carbonate (25.0g,177.0mmol) into dry N, N-dimethylformamide (100mL), replacing with nitrogen, and stirring at 100-110 ℃ overnight; after completion of the reaction, it was cooled to room temperature, extracted with pure water and ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and concentrated to obtain compound 2-2(10.0g) in yield: 72.0 percent.
MSm/z[ESI]：198.2[M+1]。1H-NMR(400MHz,CDCl3)：δ＝8.133-8.100(dd,J1＝1.2Hz,J2＝8.0Hz,1H)，7.579-7.485(m,2H)，7.286-7.245(m，1H)，3.640-3.547(m,1H)，1.412-1.396(d,J1＝6.4Hz,6H)。
Step 2: preparation of intermediate 1- (isopropylsulfonyl) -2-nitrobenzene (i.e. Compound 2-3)
To a solution of compound 2-2(13g,65.9mmol) in dichloromethane (100mL) was added m-chloroperoxybenzoic acid (25.6g,149.4mmol) at 0 ℃ with stirring, and after completion of the addition, the reaction was carried out at 0 ℃ for 16 hours; after completion of the reaction, the reaction mixture was washed with a saturated sodium bicarbonate solution, dried, concentrated, and subjected to silica gel column chromatography to obtain compound 2-3(10.6g) in yield: 70.0 percent.
MSm/z[ESI]：230.2[M+1]。1H-NMR(400MHz,CDCl3)：δ＝8.115-8.071(m,1H)，8.042-8.003(m,1H)，7.974-7.932(m,1H)，3.825-3.757(m,1H)，1.285-1.268(d,J＝6.8Hz,6H)。
And step 3: preparation of intermediate 2- (isopropylsulfonyl) aniline (Compound 2-4)
Adding compound 2-3(20.0g,87.3mmol) and 10% palladium on charcoal (2.0g) into dry methanol (250mL), replacing with nitrogen, introducing hydrogen, and reacting at 60 deg.C for 2 hr; after completion of the reaction, the reaction mixture was cooled, separated by filtration through celite, dried over anhydrous sodium sulfate, and concentrated to obtain compound 2-4(17.3g) in yield: 95.0 percent.
MSm/z[ESI]：200.2[M+1]。1H-NMR(400MHz,DMSO-d6)：δ＝7.457-7.434(m,1H)，7.373-7.300(m,1H)，6.888-6.868(d,J＝8.0Hz,1H)，6.708-6.689(d,J＝7.6Hz,1H)，6.088(m,2H)，3.370-3.304(m,1H)，1.176-1.159(d,J＝6.8Hz,6H)。
And 4, step 4: preparation of intermediate 2, 5-dichloro-N- [2- [ (1-methylethyl) sulfonyl ] phenyl ] -4-aminopyrimidine (i.e. compound 2-5)
Under the ice-water bath condition, the compound 2-4(30.0g,150mmol) is dissolved in N, N-dimethylformamide (300mL), sodium hydride (7.3g,300mmol) is slowly added into the reaction system, and stirring is carried out at 0 ℃ for 15 minutes; then dropwise adding 2,4, 5-trichloropyrimidine (33.1g,180mmol) into the reaction system, reacting at room temperature and stirring overnight; after completion of the reaction, the reaction mixture was poured into 500mL of water, extracted with ethyl acetate, dried, concentrated, and subjected to silica gel column chromatography to obtain compound 2-5(17.3g) in yield: 32.0 percent.
MSm/z[ESI]：370.2[M+1]。1H-NMR(400MHz,DMSO-d6)：δ＝9.822(s,1H)，7.343-7.323(d,J＝8.0Hz,1H)，7.911-7.839(m,2H)，3.585-3.484(m,1H)，1.176-1.159(d,J＝6.8Hz,6H)。
Synthesis scheme 3: target compound 5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4Synthesis of- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine (i.e., REX-B1)
Step 1: preparation of target Compound (i.e., REX-B1)
Dissolving compounds 1-12(400.0mg,1.31mmol), compounds 2-5(1029.0mg,2.97mmol), cesium carbonate (2.6g,8mmol), catalyst 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (232.0mg,0.4mmol) and palladium acetate (44.0mg,0.2mmol) in dioxane (15mL) and adding to the sealed tube, displacing with nitrogen, reacting at 95 ℃ for 18 hours; after the reaction was completed, the solvent was dried by evaporation, extracted with pure water and ethyl acetate, and the organic phase was dried, concentrated, and purified by silica gel column chromatography to obtain compound REX-B1(200mg) in yield: 24.8 percent.
MSm/z[ESI]：615.1[M+1]。1HNMR(400MHz,CDCl3)：δ＝9.995(s,1H)，9.152(s,1H)，8.493-8.471(d,1H,J＝8.8Hz)，8.106(s,1H)，7.974-7.954(d,1H,J＝8Hz)，7.818(s,1H)，7.563-7.519(t,1H,J＝8.8Hz)，7.359-7.321(t,1H,J＝8Hz)，7.216(s,1H)，4.723-4.684(m,1H)，3.886-3.880(m,6H)，2.725-2.718(m,5H)，2.305-2.300(s,3H)，1.361-1.266(m,16H)。
Example 25 chloro-N2- (4- (1- ((dimethylamino) methyl) cyclopropyl) -2-isopropoxy-5-methylbenzene) -N4Preparation of (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine [ numbered REX-B2 ]
The synthetic route is as follows:
Figure BDA0001042691910000161
as described in the synthetic route provided in this example, in synthesis scheme 1 "synthesis of compounds 1 to 12", dimethylamine was used in place of morpholine in example 1 in step 9, and the rest of the synthesis procedure was the same as in synthesis scheme 1 in example 1, to obtain compounds 1 to 12 in yields: 22.0 percent.
MSm/z[ESI]：263.4[M+1]。
In synthesis scheme 2 "synthesis of compound 2-5", the raw material 2-fluoronitrotoluene (i.e., compound 2-1) and the remaining synthesis methods are the same as in synthesis scheme 2 of example 1, to obtain compound 2-5 with yield: 33 percent.
MSm/z[ESI]：370.2[M+1]。
In synthesis scheme 3 "synthesis of target compound REX-B2", compounds 1 to 12 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-B2 with yield: 16 percent.
MSm/z[ESI]：573.1[M+1]。1H-NMR(400MHz,DMSO-d6)：δ＝9.484(s,1H)，9.106(s,1H)，8.430-8.409(d,1H,J＝8.4Hz)，8.3(s,1H)，8.052(s,1H)，7.903-7.883(d,1H,J＝8Hz)，7.71(s,1H)，7.452-7.414(t,1H,J＝7.6Hz)，7.02(s,1H)，4.654(m,1H)，3.456(q,1H)，3.085(s,1H)，2.327(s,1H)，2.215(s,3H)，1.284-1.076(m,18H)。
Example 35 chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((4-oxopiperidinyl) methyl) cyclopropyl) phenyl) -N4Preparation of (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine [ numbered REX-B3 ]
The synthetic route is as follows:
Figure BDA0001042691910000171
as shown in the synthetic route provided in this example, in synthetic scheme 1, since the starting materials and synthetic methods of "step 1 to step 7" in this example are the same as "step 1 to step 7" in synthetic scheme 1 in example 1, compounds 1 to 8 prepared in synthetic scheme 1 in example 1 can be directly taken and synthesized according to the route from step 8 to step 11 to obtain intermediate compounds 1 to 12, which are specifically prepared as follows:
and 8: preparation of intermediate 1- (2-methyl-4-nitro-5-isopropoxy) cyclopropylphenethyl alcohol (i.e., Compound 1-9)
Compound 1-8(500mg,1.7mmol) was dissolved in tetrahydrofuran (50ml), lithium aluminium hydride (128mg,3.37mmol) was added portionwise at 0 ℃ and the reaction heated to 25 ℃ and stirred for 14 hours; after the reaction was completed, extraction was performed with pure water and ethyl acetate, followed by filtration and concentration to obtain compounds 1 to 9(300mg) in yield: 66.67 percent.
MSm/z[ESI]：266.2[M+1]。
And step 9: preparation of intermediate 1- (1- (bromomethyl) cyclopropyl) -5-isopropoxy-2-methyl-4-nitrobenzene (i.e. compound 1-10)
Compounds 1 to 9(240.0mg,0.9mmol), triphenylphosphine (260.0mg,0.99mmol) and N-bromosuccinimide (260.0mg,0.99mmol) were dissolved in dichloromethane (50ml) and reacted at room temperature for 3 hours; after the reaction was completed, extraction was performed with pure water and ethyl acetate, and drying, spin-drying and column chromatography were performed to obtain compounds 1 to 10(238.0mg) in yield: 80.6 percent.
MSm/z[ESI]：329.4[M+1]。
Step 10: preparation of intermediate 4- ((1- (5-isopropoxy-2-methyl-4-nitrophenyl) cyclopropyl) oxomethyl) piperidine-1-carboxylic acid tert-butyl ester (i.e. Compound 1-11)
Compound 1-10(238.0mg,0.73mmol) and 1-tert-butoxycarbonyl-4-hydroxypiperidine (177.0mg,0.88mmol) were dissolved in N, N-dimethylformamide (50ml), and sodium hydride (47.0mg,1.17mmol) was added thereto at 0 ℃ to react at room temperature for 3 hours; after the reaction was completed, extraction was performed with pure water and ethyl acetate, and drying and spin-drying were performed to obtain compounds 1 to 11(140.0mg), yield: 42.8 percent.
MSm/z[ESI]：449.5[M+1]。
Step 11: preparation of intermediate t-butyl 4- ((1- (5-isopropoxy-2-methyl-4-anilino) cyclopropyl) oxomethyl) piperidine-1-carbonate (i.e. Compound 1-12)
Taking compounds 1-11, prepared according to the method of example 1, scheme 1, step 11, gives compounds 1-12 in yields: 85.0 percent.
MSm/z[ESI]：419.5[M+1]。
In synthesis scheme 2 "synthesis of compound 2-5", the raw material 2-fluoronitrobenzene (i.e., compound 2-1), and the rest of the synthesis methods are the same as in synthesis scheme 2 of example 1, to obtain compound 2-5, with yield: 33 percent.
MSm/z[ESI]：370.2[M+1]。
In synthesis scheme 3 "synthesis of target compound REX-B3", compounds 1 to 12 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-B3 with yield: 16.0 percent.
MSm/z[ESI]：629.2[M+1]。1H-NMR(400MHz，DMSO-d6)：δ＝10.028(s,1H)，9.016-8.864(m,3H)，8.485(s,1H)，8.172-8.128(d,1H,J＝17.6Hz)，7.951-7.936(d,1H,J＝6Hz)，7.775-7.735(t,1H,J＝9.2Hz)，7.568-7.531(t,1H,J＝7.6Hz)，7.357(s,1H)，6.887(s,1H),4.568-4.560(m,1H)，3.492-3.339(m,2H)，2.97-2.879(m,4H)，2.099(s,3H)，1.828(m,2H)，1.568(m,2H)，1.263-1.248(d,6H,J＝6Hz)，1.144-1.128(d,6H,J＝6.4Hz)，0.876-0.852(t,2H,J＝5.2Hz)，0.714-0.689(t,2H,J＝5.2Hz)。
Example 45 chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methanolpiperazinyl) cyclopropyl) phenyl) -N4Preparation of (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine [ numbered REX-B4 ]
The synthetic route is as follows:
Figure BDA0001042691910000181
as described in the synthetic route provided in this example, in Synthesis scheme 1, since the starting materials and the synthetic methods of "step 1 to step 8" in this example are the same as "step 1 to step 8" in Synthesis scheme 1 in example 1, Compounds 1 to 9 can be directly prepared by the method of Synthesis scheme 1 in example 1; step 9, replacing morpholine in example 1 with N-Boc-piperazine to carry out reaction to obtain compounds 1-10; step 10, reacting compounds 1-10 according to step 11 of synthesis scheme 1 of example 1 to obtain intermediate compounds 1-11, with yield: 90.0 percent.
MSm/z[ESI]：418.5[M+1]。
In synthesis scheme 2 "synthesis of compound 2-5", the raw material 2-fluoronitrobenzene (i.e., compound 2-1), and the rest of the synthesis methods are the same as in synthesis scheme 2 of example 1, to obtain compound 2-5, with yield: 33.0 percent.
MSm/z[ESI]：370.2[M+1]。
In synthesis scheme 3 "synthesis of target compound REX-B4", compounds 1 to 11 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-B4 with yield: 25.0 percent.
MSm/z[ESI]：628.2[M+1]。1H-NMR(400MHz,DMSO-d6)：δ＝9.965(s,1H)，9.305-9.300(m,2H)，8.897(s,1H)，8.474(s,1H)，8.147-8.128(d,1H,J＝7.6Hz)，7.959-7.937(d,1H,J＝7.6Hz)，7.781-7.743(t,1H,J＝7.6Hz)，7.578-7.540(t,1H,J＝8Hz)，7.44(s,1H)，7.059(s,1H)，4.722-4.720(m,1H)，3.651(s,4H)，3.471-3.470(m,1H)，2.834-2.828(m,4H)，1.963(s,3H)，1.374(s,2H)，1.255-1.241(d,6H,J＝5.6Hz)，1.139-1.123(d,6H,J＝6.4Hz)，1.037(s,2H)。
Example 55 chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((4-oxopyrrolidinyl) methyl) cyclopropyl) phenyl) -N4Preparation of (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine [ numbered REX-B29 ]
The synthetic route is as follows:
Figure BDA0001042691910000191
as described in the synthetic route provided in this example, in Synthesis scheme 1, since the starting materials and the synthetic methods of "step 1 to step 9" in this example are the same as "step 1 to step 9" in Synthesis scheme 1 in example 3, Compounds 1 to 10 can be directly prepared by the method of Synthesis scheme 1 in example 3; step 10, replacing the N-Boc-4-hydroxypiperidine in example 3 with 1-BOC-3-hydroxypyrrolidine to perform a reaction to obtain compounds 1 to 11; step 11, reacting compounds 1-11 according to step 11 of synthesis scheme 1 of example 3 to obtain intermediate compounds 1-12, with yield: 70.0 percent.
MSm/z[ESI]：405.5[M+1]。
In synthesis scheme 2 "synthesis of compound 2-5", the raw material 2-fluoronitrobenzene (i.e., compound 2-1), and the rest of the synthesis methods are the same as in synthesis scheme 2 of example 1, to obtain compound 2-5, with yield: 33.0 percent.
MSm/z[ESI]：370.2[M+1]。
In synthesis scheme 3 "synthesis of target compound REX-B29", compounds 1 to 12 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-B29 with yield: 23.0 percent.
MSm/z[ESI]：615.2[M+1]。
Example 65 chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((4-oxohydroxyazetidinyl) methyl) cyclopropyl) phenyl) -N4Preparation of (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine [ numbered REX-B30 ]
The synthetic route is as follows:
Figure BDA0001042691910000201
as described in the synthetic route provided in this example, in Synthesis scheme 1, since the starting materials and the synthetic methods of "step 1 to step 9" in this example are the same as "step 1 to step 9" in Synthesis scheme 1 in example 3, Compounds 1 to 10 can be directly prepared by the method of Synthesis scheme 1 in example 3; step 10, replacing the N-Boc-4-hydroxypiperidine of example 3 with N-Boc-3-hydroxyazetidine to perform a reaction to obtain compounds 1 to 11; in step 11, compounds 1-11 are reacted according to step 11 of synthesis scheme 1 of example 3 to give intermediate compounds 1-12, with yields: 50.0 percent.
MSm/z[ESI]：392.5[M+1]。
In synthesis scheme 2 "synthesis of compound 2-5", the raw material 2-fluoronitrobenzene (i.e., compound 2-1), and the rest of the synthesis methods are the same as in synthesis scheme 2 of example 1, to obtain compound 2-5, with yield: 33.0 percent.
MSm/z[ESI]：370.2[M+1]。
In synthesis scheme 3 "synthesis of target compound REX-B30", compounds 1 to 12 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-B30 with yield: 12.0 percent.
MSm/z[ESI]：601.2[M+1]。
Example 75 trifluoromethyl-N2- (2-isopropoxy-5-methyl-4- (1- ((4-oxopiperidinyl) methyl) cyclopropyl) phenyl) -N4The synthesis route of- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine [ numbered REX-B31 ] is as follows:
Figure BDA0001042691910000211
as described in the synthetic route provided in this example, in synthesis scheme 1 "synthesis of compounds 1 to 5", the starting material 2-phenylacetic acid (i.e., compound 1-1) and the remaining synthesis methods are the same as in synthesis scheme 1 of example 3, to obtain compounds 1 to 12 in yields: 30.0 percent.
MSm/z[ESI]：419.5[M+1]。
In synthesis scheme 2, since the raw materials and synthesis methods of "step 1 to step 3" in this example are the same as "step 1 to step 3" in synthesis scheme 2 in example 3, compound 2-4 prepared in synthesis scheme 2 in example 3 can be directly taken, and 2, 4-dichloro-5-trifluoromethylpyrimidine in step 4 is used to replace 2,4, 5-trichloropyrimidine in example 3, and then reacted to obtain compound 2-5; yield: 60.0 percent.
MSm/z[ESI]：370.2[M+1]。
In synthesis scheme 3 "synthesis of target compound REX-B31", compounds 1 to 12 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-B30 with yield: 12.0 percent.
MSm/z[ESI]：663.2[M+1]。1HNMR(400MHz,DMSO),δ:9.523(s,1H),9.279-9.228(m,2H),8.456(s,1H),8.222(m,2H),7.821-7.801(d,1H,J＝8),7.628(s,1H),7.559-7.524(t,1H,J＝7),7.360-7.322(t,1H,J＝7.6),6.946(s,1H),4.580-4.523(m,1H),3.477-3.429(m,3H),2.960-2.860(m,4H),2.227(s,3H),1.846(m,2H),1.608(m,2H),1.191-1.176(d,6H,J＝6),1.147-1.131(d,6H,J＝6.4),0.892(s,2H),0.732(s,2H)。
Example 8 measurement of ALK kinase inhibitory Activity and binding Rate to related mutation sites
For the compounds REX-B1-REX-B4 prepared in examples 1-4, the following compounds were used
Figure BDA0001042691910000222
(FRET) method for determining ALK kinase inhibitory activity of the aforementioned compound, the inhibitory activity being IC50Expressed by this index, IC50I.e., the concentration of the compound at which the activity of ALK kinase is inhibited by 50%.
At the same time adopt
Figure BDA0001042691910000223
Eu Kinase binding Assay (TR-FRET) Assay for determining the binding rate of the compounds of the present invention to ALK-associated mutation sites, such as ALK L1196M, and also using IC50This index is used to express. Lantha screen Eu kinase binding assay Alexa Fluor conjugate or kinase "tracer" binding was detected by addition of Eu-labeled antibody or anti-tag antibody. Binding of the tracer and antibody to the kinase results in a high degree of FRET, whereas the use of a kinase inhibitor instead of the tracer results in loss of FRET.
The invention utilizes the kinase assay platform of Life technology company to carry out the determination, and the determination result is shown in the table I. The result shows that the compound provided by the invention has better ALK inhibitory activity and better binding rate to ALK mutation sites (such as ALK L1196M).
TABLE ALK inhibitory Activity and ALK L1196M binding Rate determination of the Compounds of the examples
ALK IC50(nM) ALK F1196M IC50(nM)
REX-B1 143 121
REX-B2 <30 <30
REX-B3 <30 <30
REX-B4 <100 <30
Further, the compounds REX-B1, REX-B2, REX-B3 and REX-B4 selected by the present invention were tested for their activity against the relevant kinases at a concentration of 100nM using the kinase assay platform from Life technology, Inc., and the results are shown in Table two. The results show that the compounds REX-B2, REX-B3 and REX-B4 provided by the invention show good binding rate to ALK F1174L and ALK C1156Y, and show high inhibitory activity to LTK and ROS 1.
TABLE II example determination of the Activity of Compounds on the relevant kinases at a concentration of 100nM
Figure BDA0001042691910000221
Example 9 cell proliferation assay (MTT assay)
Test compounds: the compounds REX-B2, REX-B3 and REX-B4 prepared in the embodiments 2-4 of the invention.
Cell lines: lung cancer cell lines NCI-H2228 and NCI-H3122 were purchased from Bai Biotech limited of Nanjing.
The method comprises the following steps: the cell lines NCI-H2228 and NCI-H3122 were cultured in 20% FBS (fetal bovine serum) (Gibco) +1640+ 1% double antibody. Then taking NCI-H2228 and NCI-H3122 cells with good growth state, inoculating 5000 cells/well into 96-well cell plate, placing at 37 deg.C and containing 5% CO2The culture box is incubated for 24 hours to ensure that the cells are completely attached. Discarding old culture solution, sequentially adding 100 μ L culture solution containing 0.3, 1, 3, 10, 30, 100, 300, 1000, 3000 and 10000nmol/L compounds to be tested into each well, adding 100 μ L culture solution containing 0.1% DMSO into each well of solvent control group, repeating 3 wells for each group, discarding old culture solution after 72 hr, adding 100 μ L culture solution containing 0.5 mg/mL into each well under dark condition-1And (3) placing the MTT culture solution in a cell culture box, continuously incubating for 4h, removing the supernatant, adding 100 mu L DMSO into each hole, oscillating, and measuring the absorbance value of each hole by using a microplate reader at 490nm wavelength. The IC of each compound on each cell was calculated using graphad 6.0 based on the inhibition of each cell growth by different concentrations of each compound50The value is obtained.
The calculation formula is as follows:
Figure BDA0001042691910000231
as a result: see table three.
Detection of cell proliferation IC of example compound on lung cancer cell line by using epi-three MTT method50Value of
IC50(μM) NCI-H2228 NCI-H3122
REX-B2 <1 <1
REX-B3 <1 <1
REX-B4 <1 <1
Example 10 cell proliferation assay (CKK-8 assay)
Test compounds: compounds REX-B1 and REX-B2 prepared in examples 1-2 of the present invention.
Cell lines: the human-induced degeneration large cell lymphoma cell strain Karpas-299 is purchased from Bai Biotech limited company of Nanjing Kelvin.
The method comprises the following steps: cells in the logarithmic growth phase were collected, counted, resuspended in complete medium, adjusted to the appropriate concentration (as determined by the cell density optimization assay) and seeded into 96-well plates with 100. mu.l of cell suspension per well. Cells were incubated at 37 ℃ in a 100% relative humidity, 5% CO2 incubator for 24 hours. Combining the compounds to be tested with a culture mediumThe material was diluted to the corresponding working concentration set and cells were added at 25. mu.l/well. The final concentration of compound was started at 10. mu.M, and diluted in 3-fold gradient for 10 concentration points. Cells were incubated at 37 ℃ and 100% relative humidity, 5% CO2Incubate in incubator for 72 hours. The solution of CCK-8 with 1/10 volume is directly added, mixed evenly and put in an incubator at 37 ℃ for 1 to 4 hours. After gentle shaking, absorbance at a wavelength of 450nm was measured on a SpectraMax M5Microplate Reader, and the inhibition rate was calculated with the absorbance at 650nm as a reference. The IC of each compound on each cell was calculated using graphad 6.0 based on the inhibition of each cell growth by different concentrations of each compound50The value is obtained.
The calculation formula is as follows:
Figure BDA0001042691910000232
as a result: see table four.
TABLE IV CKK-8 method for detecting cell proliferation IC of example compound to Karpas-299 cell strain50Value of
IC50(μM) Karpas-299
REX-B1 <1
REX-B2 <0.05
Example 11 Zebra fish phenotype screening experiment
The zebra fish is a vertebrate, has 85 percent of gene homology with human, has a signal transduction path basically similar to that of the human, and has a biological structure and a physiological function which are highly similar to those of mammals; the zebra fish is small in size, rapid in development, transparent in embryo and high in egg laying amount, and the unique advantages enable the zebra fish to become one of the best model organisms for human disease research and living high-throughput drug screening. Among them, anaplastic lymphoma kinase ALK (anaplastic lymphoma kinase) has 76% of gene homology with zebrafish in human. The leucocyte tyrosine kinase ltk (leucocyte tyrosine kinase) of the zebra fish regulates and controls the generation of the pigmented cells of the iris of the zebra fish (Lopes, S.S., Yang, X., et al (2008). Leucocyte tyrosine kinase functions in pigment cell definition. PLoS Gene, 4.), the pigmented cells of the iris are expressed as a kind of silver corpuscle in the zebra fish and are distributed at the outer side of the head, eyes and spine (observed by reflected light), and the silver corpuscle is observed to be black in the transmitted light by utilizing the albono zebra fish with the loss of the melanin, which is shown in figure 1. ALK and LTK are sitter kinase, researchers found that injection of exogenous ALK plasmid also regulated iridescent pigment cell production, and experimental results also showed that most ALK inhibitors had LTK activity that inhibited iridescent pigment cell production (Rodrigues, F.S., Yang, X., Nikaido, M., Liu, Q., & Kelsh, R.N. (2012). A simple, highly visual in vivo screen for and adaptive fluorescence kinase inhibition. ACS Chem Biol,7, 1968-.
Therefore, by using this principle, we examined the effect of the compound on normal zebra fish iris pigmented cells to investigate the strength of the anti-ALK activity of the compound in vivo.
Effect of the test Compound (I) on Normal Zebra Fish iridescent pigmented cells
The scheme is as follows: selecting 6hpf (waters post fertilization) roes, randomly grouping, adding tested drugs with various concentrations, carrying out image acquisition when the volume reaches 3dpf (days post fertilization), analyzing IOD (integrated optimization) value of iris pigmented cells on the dorsal side of a zebra fish spine from a cloaca to a caudal fin part by using ImageJ software, carrying out Dunnett's T-test by using Graphpad prism6.0 for statistical analysis, wherein p <0.05 shows that the statistical difference exists, and the calculation formula of the iridoid inhibition rate is as follows:
Figure BDA0001042691910000241
the calculation result is shown in the fifth table, the dose-effect relationship graph of the effect of the compounds REX-B1/REX-B2 on the pigmented cells of the iris of the zebra fish is shown in figure 2, and the effect graph of the compounds REX-B1/REX-B2 on the pigmented cells of the iris of the zebra fish is shown in figure 3.
TABLE penta-compound effects on pigmented cells of iris membranes of zebra fish (mean. + -. sem)
Figure BDA0001042691910000251
compared with control,*,p<0.05；**,p<0.01。

Claims (8)

1. A4-saturated cyclic group substituted aniline protein kinase inhibitor is a compound with the following structural general formula I and pharmaceutically acceptable salts thereof:
Figure FDA0003506054350000011
wherein R is1Is selected from C1-6Alkyl, heterocyclic radical, - (CH)2)wOR10、-(CH2)wNR10R11、-CO2R10、-CONR10R11、-(CH2)wSO2R8One or more of the above;
R2is selected from C1-6An alkyl group;
R4is selected from C1-6An alkoxy group;
R3、R5each independently selected from hydrogen;
R6selected from halogens;
R7is selected from-SO2R8；
R8Is selected from C1-6An alkyl group;
R10、R11selected from hydrogen, C1-6Alkyl radicalOr a heterocyclic group;
the heterocyclic group is a 3-6 membered heterocyclic ring containing N and/or O heteroatoms;
and n is selected from 1 and w is selected from 0 or 1.
2. A4-saturated cyclic group substituted aniline protein kinase inhibitor is selected from the following characteristic compounds:
5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methylenemorpholinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
5-chloro-N2- (4- (1- ((dimethylamino) methyl) cyclopropyl) -2-isopropoxy-5-methylbenzene) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- ((4-oxopiperidinyl) methyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine;
5-chloro-N2- (2-isopropoxy-5-methyl-4- (1- (methanolpiperazinyl) cyclopropyl) phenyl) -N4- (2- (isopropylsulfonyl) phenyl) -2, 4-diaminopyrimidine.
3. A pharmaceutical composition comprising a compound as defined in any one of claims 1 or 2, or a pharmaceutically acceptable salt thereof, as active ingredient, together with one or more pharmaceutically acceptable carriers.
4. Use of a compound as defined in any one of claims 1 or 2, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment or prevention of a disease associated with a protein kinase.
5. Use of a compound as defined in any one of claims 1 or 2, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment or prevention of a disease associated with anaplastic lymphoma kinase.
6. Use according to claim 5, characterized in that: the disease is selected from cell proliferative diseases.
7. Use according to claim 6, characterized in that: the disease is selected from tumors.
8. Use according to claim 7, characterized in that: the tumor is selected from non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastoma, nasopharyngeal carcinoma, breast cancer, colorectal cancer, diffuse large B cell lymphoma, liver cancer, gastric cancer, esophageal cancer, pancreatic cancer, ovarian cancer, systemic histiocytosis or neuroblastoma.
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