CN106146468B - Pyridone protein kinase inhibitors - Google Patents

Pyridone protein kinase inhibitors Download PDF

Info

Publication number
CN106146468B
CN106146468B CN201510183879.2A CN201510183879A CN106146468B CN 106146468 B CN106146468 B CN 106146468B CN 201510183879 A CN201510183879 A CN 201510183879A CN 106146468 B CN106146468 B CN 106146468B
Authority
CN
China
Prior art keywords
compound
protein kinase
rex
amino
cancer
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201510183879.2A
Other languages
Chinese (zh)
Other versions
CN106146468A (en
Inventor
王永辉
高羽军
周娟
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hangzhou REX Pharmaceutical Co.,Ltd.
Original Assignee
Hangzhou Rex Pharmaceutical Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hangzhou Rex Pharmaceutical Co ltd filed Critical Hangzhou Rex Pharmaceutical Co ltd
Priority to CN201510183879.2A priority Critical patent/CN106146468B/en
Publication of CN106146468A publication Critical patent/CN106146468A/en
Application granted granted Critical
Publication of CN106146468B publication Critical patent/CN106146468B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • 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
    • C07D401/14—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 three or more hetero rings
    • 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
    • C07D405/14—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 three or more hetero rings

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 pyridone 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

Pyridone protein kinase inhibitors
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 pyridone 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 pyridone protein kinase inhibitor with a novel structure, which synthesizes and screens a series of compounds with anti-tumor activity (having inhibition effect on EML4-ALK positive and secondary mutation) by carrying out substitution modification on a pyridone group.
In order to realize the purpose, the invention adopts the following technical scheme:
a pyridone protein kinase inhibitor is a compound with the following structural formula and pharmaceutically acceptable salts thereof:
Figure BDA0000701187500000021
R1selected from hydrogen,
Figure BDA0000701187500000022
R0、R2、R3、R4、R11Each independently selected from hydrogen, halogen, C1-6Alkyl, halo C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy radical, C1-6Cycloalkyl, halo C1-6Cycloalkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, cyano or amino; r5Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, C1-6Alkoxy, halo C1-6Alkoxy, -NR2COR6、-NR2CONR2R6、-NR2SO2R6、-COR6、-CONR2R6、-SO2R6、-SO2NR2R6、-POR8R9、
Figure BDA0000701187500000023
R6Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6One or more of alkynyl, hydroxyl and aryl, or
Figure BDA0000701187500000024
R7Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, C1-6Alkoxy, hydroxyl, aryl, one or more of 3-12 membered heterocyclic radical with N, O hetero atoms, or
Figure BDA0000701187500000031
R8、R9Each independently selected from hydrogen and C1-6Alkyl radical, C2-6Alkenyl radical, C2-6One or more of alkynyl, hydroxyl and aryl; r10Selected from hydrogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, amido, hydroxyl, aryl, and one or more of 3-12 membered heterocyclic group with N, O heteroatoms; and m, n, p are each independently selected from any integer value from 0 to 10.
Preferably, the aryl is one or more of phenyl, naphthyl and anthryl; the 3-12 membered heterocyclic group of the N, O heteroatom is one or more of pyridyl, piperidyl, pyrimidyl, furyl and morpholinyl.
Preferably, the halogen is one or more of fluorine, chlorine, bromine and iodine.
More preferably, R in the general structural formula as described above1Is selected from
Figure BDA0000701187500000032
R0、R2、R3、R4、R11Each independently selected from hydrogen, halogen, C1-6Alkyl, halo C1-6Alkyl radical, C1-6Alkoxy radicalRadical, halo C1-6Alkoxy radical, C1-6Cycloalkyl, halo C1-6Cycloalkyl, cyano or amino; r5Selected from hydrogen, C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy, SO2R6、POR8R9、
Figure BDA0000701187500000033
R6Selected from hydrogen, C1-6Alkyl radical, C1-6Alkyl hydroxy or
Figure BDA0000701187500000034
R7Selected from hydrogen, C1-6Alkoxy, halo C1-6Alkoxy, a 3-to 6-membered heterocycle from the group consisting of the heteroatoms N, O or
Figure BDA0000701187500000035
R8、R9Each independently selected from hydrogen or C1-6An alkyl group; r10Selected from hydrogen, C1-6Alkyl radical, C1-6Alkylamide group, C1-6An alkylhydroxy group or a 3-6 membered heterocyclic ring selected from the group consisting of the heteroatoms N, O; and m, n, p are each independently selected from any one of the values 0, 1, 2, 3, 4,5, 6.
A pyridone protein kinase inhibitor, which is selected from the following characteristic compounds numbered REX-1-REX-23:
REX-1: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-2: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-methylenepiperidinyl) pyridin-2 (1 h) -one;
REX-3: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -5-methyl-1- (4-methylenepiperidinyl) pyridin-2 (1 h) -one;
REX-4: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -5-methyl-1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-5: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -6-methyl-1- (4-methylenepiperidinyl) pyridin-2 (1 h) -one;
REX-6: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -6-methyl-1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-7: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -5-fluoro-1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-8: 5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-9: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) -5-trifluoromethylpyridin-2 (1 h) -one;
REX-10: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) -5-trifluoromethylpyridin-2 (1 h) -one;
REX-11: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -5-isopropoxy-1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-12: 2- (4- (4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -2-1(2H) pyridonyl) -1-piperidine) acetamide;
REX-13: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (1- (2-hydroxyethyl) -4-piperidinyl) pyridin-2 (1 h) -one;
REX-14: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4- (1-ethylpiperidinyl)) pyridin-2 (1 h) -one;
REX 15: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4- (1-2H-tetrahydropyranyl) piperidinyl) pyridin-2 (1H) -one;
REX-16: 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-2 hydro-pyranyl) pyridin-2 (1 hydro) -one;
REX-17: 4- ((4- ((2- (isopropylsulfonyl) phenyl) amine-5-trifluoromethyl) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-18: 4- ((5-chloro-4- ((2- (phosphodimethyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-19: 4- ((5-chloro-4- ((2- (N, N dimethylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-20: 4- ((5-chloro-4- ((2- (N-isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-21: 4- ((5-chloro-4- ((2- (hydroxyethylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-22: 4- ((5-chloro-4- ((2- (1- (ethoxymethyl) cyclopropyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one;
REX-23: 4- ((5-chloro-4- ((2-methoxyphenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one.
The structural formula of the compounds numbered REX-1-REX-23 is shown as follows:
Figure BDA0000701187500000051
Figure BDA0000701187500000061
the present invention also provides a process for the preparation of a compound as hereinbefore described, the general reaction scheme being as follows:
Figure BDA0000701187500000062
based on the general reaction route, the method comprises the following synthetic schemes:
(1) synthesis scheme 1: synthesis of Compounds 1-3
Step 1: r is to be1dissolving-OH (namely the compound 1-1) and triethylamine in an organic solvent, and slowly dropwise adding methylsulfonyl chloride; after reacting for N hours at normal temperature, adding an extracting agent, extracting, drying, decompressing and spin-drying to obtain the compound 1-2.
Step 2: dissolving 4-amino-2-hydroxypyridine compounds in an organic solvent, slowly adding the compound 1-2, heating for reacting 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, preferably dichloromethane, N-dimethylformamide or a mixture thereof; the extractant is selected from one or more of dichloromethane, pure water and ethyl acetate, preferably dichloromethane, pure water or their mixture;
in the synthesis scheme 1, the reaction temperature at normal temperature is 20-30 ℃, and 25 ℃ is preferred; the heating reaction temperature is 70-90 ℃, preferably 80 ℃, and the reaction time N is 3-10 hours.
(2) Synthesis scheme 2: synthesis of Compounds 2-4
Step 1: dissolving 2-fluoro-nitrobenzene and R5-H in an organic solvent, slowly adding potassium carbonate, heating for reacting for N hours, adding an extracting agent, extracting, drying, decompressing, and spin-drying to obtain a compound 2-2.
Step 2: dissolving the compound 2-2 in an organic solvent, adding a catalyst, replacing nitrogen, introducing hydrogen, heating for reacting for N hours, filtering under reduced pressure to remove the redundant catalyst, and carrying out reduced pressure and spin drying to obtain the compound 2-3.
And step 3: dissolving the compound 2-3 in an organic solvent, slowly adding sodium hydride and 2,4, 5-trichloropyrimidine, displacing with nitrogen, heating for reacting for N hours, adding a small amount of ice water to destroy redundant sodium hydride, adding an extracting agent, extracting, drying, reducing pressure and spin-drying to obtain the compound 2-4.
In the synthesis scheme 2, the organic solvent is selected from one or more of dichloromethane, N-dimethylformamide, methanol and dioxane, preferably N, N-dimethylformamide, methanol or a mixture thereof; the extractant is selected from one or more of dichloromethane, pure water and ethyl acetate, preferably ethyl acetate, pure water or their mixture; the catalyst is selected from one or more of palladium carbon, palladium acetate and 4, 5-bis (diphenylphosphino) -9, 9-dimethyl xanthene, and preferably 10% of palladium carbon.
In the synthesis scheme 2, the heating reaction temperature is 60-120 ℃, preferably 60 ℃, 100 ℃ or 120 ℃; the reaction time N is 12 to 16 hours, preferably 12 hours, 14 hours or 16 hours.
(3) Synthesis scheme 3: synthesis of target Compound
Step 1: dissolving the compounds 1-3 and 2-4 in organic solvent, adding cesium carbonate and catalyst, nitrogen replacing, microwave heating for N hr, filtering under reduced pressure to remove excessive catalyst, and performing column chromatography to obtain the target compound.
In the synthesis scheme 3, the organic solvent is selected from one or more of dichloromethane, N-dimethylformamide, methanol and dioxane, preferably dioxane; the catalyst is selected from one or more of palladium carbon, palladium acetate and 4, 5-bis-diphenylphosphine-9, 9-dimethyl xanthene, and preferably the mixture of palladium acetate and 4, 5-bis-diphenylphosphine-9, 9-dimethyl xanthene.
In the synthesis scheme 3, the reaction time N is 0.5 to 1 hour, preferably 0.5 hour.
In schemes 1, 2 and 3, R1Selected from hydrogen,
Figure BDA0000701187500000081
R0、R2、R3、R4、R11Each independently selected from hydrogen, halogen, C1-6Alkyl, halo C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy radical, C1-6Cycloalkyl, halo C1-6Cycloalkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, cyano or amino; r5Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, C1-6Alkoxy, halo C1-6Alkoxy, -NR2COR6、-NR2CONR2R6、-NR2SO2R6、-COR6、-CONR2R6、-SO2R6、-SO2NR2R6、-POR8R9、
Figure BDA0000701187500000082
Figure BDA0000701187500000083
R6Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6One or more of alkynyl, hydroxyl and aryl, or
Figure BDA0000701187500000084
R7Selected from hydrogen, halogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, C1-6Alkoxy, hydroxyl, aryl, one or more of 3-12 membered heterocyclic radical with N, O hetero atoms, or
Figure BDA0000701187500000085
R8、R9Each independently selected from hydrogen and C1-6Alkyl radical, C2-6Alkenyl radical, C2-6One or more of alkynyl, hydroxyl and aryl; r10Selected from hydrogen, C1-6Alkyl radical, C2-6Alkenyl radical, C2-6Alkynyl, amido, hydroxyl, aryl, and one or more of 3-12 membered heterocyclic group with N, O heteroatoms; and m, n, p are each independently selected from any integer value from 0 to 10.
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 "carbonyl" refers to-CO.
The term "carboxyl" refers to-COOH.
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 BDA0000701187500000091
Specific examples include, but are not limited to-NHCH3、-NHCH(CH3)2、-NHC2H5Etc.; -N (alkyl)2In the structural form of
Figure BDA0000701187500000092
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 heteroaryl groups include, but are not limited to, pyrrolyl, indolyl, pyrrolidinyl, imidazolyl, pyrazolyl, tetrazolyl, pyridyl, quinolyl, isoquinolyl, piperidyl, pyrimidinyl, pyrazinyl, piperazinyl, furyl, 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 an activity inhibition experiment on a series of synthesized pyridone protein kinase inhibitor compounds on ALK kinase and ALK related mutation sites, and finds that part of the compounds have significant activity inhibition on the ALK kinase and the ALK mutation site L1196M; in addition, cell proliferation experiments of lung cancer cell lines are also carried out, and partial compounds are found to have remarkable inhibitory effects.
Compared with the prior art, the pyridone protein kinase inhibitor provided by the invention is based on the reasonable drug design of a target, and a series of compounds with novel structures are obtained by substitution modification on a pyridone group; and a series of compounds with anti-tumor activity (especially having inhibitory effect on EML4-ALK positive and secondary mutation) are optimized and screened. 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.
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 preparation of- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one [ number REX-1 ]
The synthetic route is as follows:
Figure BDA0000701187500000131
synthesis scheme 1: synthesis of intermediate t-butyl 4- (4-amino-2-oxopyridin-1 (2h) yl) piperidinyl-1-carboxylate (i.e. Compound 1-4)
Step 1: preparation of intermediate N-tert-butyloxycarbonyl-4-hydroxypiperidine (i.e., compound 1-2)
The starting material, 4-hydroxypiperidine (compound 1-1, 10g,99.07mmol), was dissolved in methylene chloride (100mL), and di-tert-butyl dicarbonate (21.7g,99.1mmol) was added to the reaction mixture, followed by stirring at room temperature for 2 to 4 hours. After the reaction, methyl t-butyl ether (200mL) and 0.5N hydrochloric acid (200mL) were added and extracted. The organic phase was dried over anhydrous sodium sulfate and concentrated to give compound 1-2(15.0g), yield: 75.3 percent. MSm/z [ ESI ]: 202.1[ M +1 ].
Step 2: preparation of intermediate 4-methanesulfonyloxypiperidine-1-carboxylic acid tert-butyl ester (i.e., compound 1-3)
Compound 1-2(15.0g,74.6mmol) and triethylamine (21mL,153mmol) were dissolved in dichloromethane (200mL), methanesulfonyl chloride (6.0mL,74.6mmol) was slowly added dropwise at 5 ℃ or lower, and the temperature was maintained at 5-15 ℃ for 30 minutes. After the reaction was completed, the temperature was returned to room temperature, and the mixture was diluted with 400mL of water and extracted with methylene chloride. The organic phase was washed twice with sodium bicarbonate, dried over anhydrous sodium sulfate, and concentrated to give compound 1-3(13.0g), yield: 62.0 percent. MSm/z [ ESI ]: 280.1[ M +1 ].
And step 3: preparation of intermediate tert-butyl 4- (4-amino-2-oxopyridin-1 (2 hydro) yl) piperidinyl-1-carboxylate (i.e. Compound 1-4)
4-amino-2-hydroxypyridine (5.1g,46.6mmol) was dissolved in DMF (80mL) in an ice-water bath, and sodium hydride (2.0g,51.3mmol) was slowly added to the reaction system and stirred at 0 ℃ for 15 minutes. Then, compound 1-3(13.0g,46.6mmol) was added to the reaction system, and the mixture was heated to 45 ℃ and stirred overnight. 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 compound 1-4(6.0g) in yield: 43.9 percent. MSm/z [ ESI ]]：294.4[M+1]。1H-NMR(400MHz，DMSO6)：＝7.324-7.305(d，J＝7.6Hz，1H)，5.963(s，2H)，5.694-5.669(m，1H),5.246-5.240(d，J＝2.4Hz，1H),4.726(s，1H)，4.099-4.058(m，2H),2.519-2.501(m，2H),1.623-1.590(m，4H)，1.419-1.388(s，9H)。
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)
Raw materials 2-fluoronitrobenzene (compound 2-1, 10g,70mmol), isopropyl mercaptan (5.4g,70mmol) and potassium carbonate (25g,177mmol) were added to dry DMF (100mL), replaced with nitrogen, and stirred overnight at 100-110 ℃. The reaction mixture was cooled to room temperature, water (200mL) was added, and extraction was performed with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to give compound 2-2(10g), yield: 72 percent. MSm/z [ ESI ]]：198.2[M+1]。1H-NMR(400MHz，CDCl3)：＝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,65.97mmol) in dichloromethane (100mL) was added m-CPBA (25.67g,149.42mmol) in portions with stirring at 0 ℃ and, after the addition, reacted at 0 ℃ for 16 hours. The reaction mixture was washed with saturated sodium bicarbonate solution, dried, concentrated and subjected to silica gel column chromatography to obtain compound 2-3(10.66g) in yield: 70 percent. MSm/z [ ESI ]]：230.2[M+1]。1H-NMR(400MHz，CDCl3)：＝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)
Compound 2-3(20g,87.3mmol) and 10% palladium on charcoal (2.0g) were added to dry methanol (250mL), displaced with hydrogen, and reacted at 60 ℃ for 2 hours. After cooling, the mixture was separated by filtration through celite, and dried and concentrated over anhydrous sodium sulfate to obtain compound 2-4(17.3g) in yield: 95 percent. MSm/z [ ESI ]]：200.2[M+1]。1H-NMR(400MHz，DMSO6)：＝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 ice-water bath conditions, compounds 2-4(30g,150mmol) were dissolved in DMF (300mL), and sodium hydride (7.23g,300mmol) was slowly added to the reaction system and stirred at 0 ℃ for 15 minutes. 2,4, 5-trichloropyrimidine (33.1g,180mmol) was added dropwise to the reaction system, and the reaction was stirred at room temperature overnight. 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 compound 2-5(17.3g) in yield: 32 percent. MSm/z [ ESI ]]：370.2[M+1]。1H-NMR(400MHz，DMSO6)：＝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: synthesis of target compound 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one (i.e., REX-1)
Step 1: preparation of intermediate tert-butyl 4- (4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -2-oxopyridin-1 (2h) -yl) piperidine-1-carboxylate (i.e. compound 3-1)
Compound 1 to 4(0.7g,2.6mmol), compound 2 to 5(0.91g,2.38mmol), cesium carbonate (2.34g,7.17mmol), catalyst 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (0.3g,0.5mmol) and palladium acetate (54mg,0.24mmol) were dissolved in dioxane (15mL) and charged into a sealed tube, and after replacing the air with nitrogen, the reaction was carried out at 95 ℃ for 18 hours. Spin-drying the solvent, adding ethyl acetate and water, extracting, drying the organic phase, and purifying by silica gel column chromatography to obtain compound 3-1(0.7g), yield: 52 percent. MSm/z [ ESI ]: 603.1[ M +1 ].
Step 2: preparation of target compound 4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-piperidinyl) pyridin-2 (1 h) -one (i.e. REX-1)
Compound 3-1(0.5g, 8.3mmol) was added to dichloromethane (10mL) with stirring, trifluoroacetic acid (5mL) was added dropwise, and the mixture was stirred at room temperature overnight. Firstly, adjusting the pH value to be more than or equal to 10 by using a 5% sodium bicarbonate aqueous solution, then extracting by using ethyl acetate, drying and spin-drying to obtain a target compound REX-1, wherein the yield is as follows: 36 percent. MSm/z [ ESI ]]：504.1[M+1]。1H-NMR(400MHz，DMSO6)：＝9.741(s，1H)，8.575-8.555(d，J＝8Hz，1H)，8.392(s，1H)，7.875-7.806(m，2H)，7.507-7.391(m，2H)，6.940-6.936(d，J＝1.6Hz，1H)，6.493-6.469(m，1H)，4.667(m，1H)，3.572-3.441(m，1H)，3.059-3.028(m，2H)，2.599-2.583(m，2H)，1.648-1.601(m，4H)，1.231-1.116(m，6H)。
Example preparation of- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -1- (4-methylenepiperidinyl) pyridin-2 (1 h) -one [ number REX-2 ]
The synthetic route is as follows:
Figure BDA0000701187500000161
as described in the synthetic route provided in this example, in synthesis scheme 1 "synthesis of compounds 1 to 4", the starting material 4-hydroxypiperidine (i.e., compound 1-1) in example 1 is replaced with 4-hydroxymethylpiperidine, and the rest of the synthesis procedure is the same as in synthesis scheme 1 in example 1, to obtain compounds 1 to 4 in yields: 43.0 percent. MSm/z [ ESI ]: 308.4[ 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-2", compounds 1 to 4 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-2 with yield: 36 percent. MSm/z [ ESI ]]：518.1[M+1]。1H-NMR(400MHz，DMSO6)：＝9.689(s，1H)，8.540-8.364(m，2H)，7.868-7.779(m，2H)，7.412(m，2H)，6.916(m，1H)，6.423(m，1H)，4.667(m，1H)，3.644-3.635(m，3H)，2.910(m，2H)，2.355(m，2H)，1.800(m，1H)，1.435(m，2H)，1.170-1.163(m，8H)。
Example preparation of 34- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -5-methyl-1- (4-methylenepiperidinyl) pyridin-2 (1 h) -one [ code REX-3 ]
The synthetic route is as follows:
Figure BDA0000701187500000171
as shown in the synthetic route provided in this example, in synthesis scheme 1 "synthesis of compounds 1 to 4", the starting material 4-hydroxypiperidine (i.e., compound 1-1) in example 1 is replaced with 4-hydroxymethylpiperidine, 4-amino-2-hydroxypyridine in step 3 in example 1 is replaced with 4-amino-5-methyl-2-hydroxypyridine, and the rest of the synthesis procedure is the same as in synthesis scheme 1 in example 1, whereby compounds 1 to 4 are obtained in yields: 38.0 percent. MSm/z [ ESI ]: 322.4[ 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: 34 percent. MSm/z [ ESI ]: 370.2[ M +1 ].
In synthesis scheme 3 "synthesis of target compound REX-3", compounds 1 to 4 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-3 in yield: 26 percent. MSm/z [ ESI ]]：532.1[M+1]。1H-NMR(400MHz，DMSO6)：＝8.646-8.625(d，J＝8.4Hz，1H)，8.625(s，1H)，8.371(s,1H)，7.845-7.824(d，J＝8.4Hz，1H)，7.733(m,1H),7.373(m，2H)，6.961(s，1H),3.351-3.663(d，J＝7.2Hz，2H)，3.535(m，3H)，2.976-2.943(d，J＝13.2Hz，2H)，2.433(m，3H)，2.039(s，3H)，1.836(m，1H)，1.447(m，2H)，1.181(m，6H)。
Example preparation of- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -5-methyl-1- (4-piperidinyl) pyridin-2 (1 h) -one [ number REX-4 ]
The synthetic route is as follows:
Figure BDA0000701187500000181
as described in the synthetic route provided in this example, in synthesis scheme 1 "synthesis of compounds 1 to 4", 4-amino-2-hydroxypyridine of step 3 in example 1 was replaced with 4-amino-5-methyl-2-hydroxypyridine, and the rest of the synthesis method was the same as in synthesis scheme 1 of example 1, to obtain compounds 1 to 4 in yields: 35.0 percent. MSm/z [ ESI ]: 307.4[ 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: 31 percent. MSm/z [ ESI ]: 370.2[ M +1 ].
In synthesis scheme 3 "synthesis of target compound REX-4", compounds 1 to 4 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-4 with yield: 30 percent. MSm/z [ ESI ]]：516.1[M+1]。1H-NMR(400MHz，DMSO6)：＝8.662-8.641(d，J＝8.4Hz，1H)，8.447(s，1H)，8.386(s，1H)，7.851-7.831(d，J＝8.0Hz，1H)，7.754(m，1H)，7.433(s，1H)，7.391(m，1H)，6.974(s，1H)，4.709(m，1H)，4.041(m，1H)，3.503(m，3H)，3.076(m，2H)，2.607(m，2H)，2.078(s，3H)，1.992(m，2H)，1.712(m，4H)，1.184(m，6H)。
Example preparation of 54- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -6-methyl-1- (4-piperidinyl) pyridin-2 (1 h) -one [ code REX-5 ]
The synthetic route is as follows:
Figure BDA0000701187500000191
as shown in the synthetic route provided in this example, in synthesis scheme 1 "synthesis of compounds 1 to 4", the starting material 4-hydroxypiperidine (i.e., compound 1-1) in example 1 is replaced with 4-hydroxymethylpiperidine, 4-amino-2-hydroxypyridine in step 3 in example 1 is replaced with 4-amino-6-methyl-2-hydroxypyridine, and the rest of the synthesis procedure is the same as in synthesis scheme 1 in example 1, whereby compounds 1 to 4 are obtained in yields: 45.0 percent. MSm/z [ ESI ]: 321.4[ 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: 38 percent. MSm/z [ ESI ]: 370.2[ M +1 ].
In synthesis scheme 3 "synthesis of target compound REX-5", compounds 1 to 4 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-5 in yield: 26 percent. MSm/z [ ESI ]]：531.1[M+1]。1H-NMR(400MHz，DMSO6)：＝8.650-8.629(d，J＝8.4Hz，1H)，8.620(s，1H)，8.370(s，1H)，7.835-7.814(d，J＝8.4Hz，1H)，7.722(m，1H)，7.352(m，2H)，6.955(s，1H)，3.351-3.663(d，J＝7.2Hz，2H)，3.535(m，3H)，2.976-2.944(d，J＝13.1Hz，2H)，2.430(m，3H)，2.239(s，3H)，1.836(m，1H)，1.447(m，2H)，1.181(m，6H)。
Example preparation of 64- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -6-methyl-1- (4-piperidinyl) pyridin-2 (1 h) -one [ code REX-6 ]
The synthetic route is as follows:
Figure BDA0000701187500000201
Figure BDA0000701187500000211
as described in the synthetic route provided in this example, in synthesis scheme 1 "synthesis of compounds 1 to 4", 4-amino-2-hydroxypyridine in step 3 in example 1 was replaced with 4-amino-6-methyl-2-hydroxypyridine, and the rest of the synthesis method was the same as in synthesis scheme 1 of example 1, to obtain compounds 1 to 4 in yields: 36.0 percent. MSm/z [ ESI ]: 308.4[ 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: 38 percent. MSm/z [ ESI ]: 370.2[ M +1 ].
In synthesis scheme 3 "synthesis of target compound REX-6", compounds 1 to 4 and compounds 2 to 5 prepared in this example were prepared according to synthesis scheme 3 of example 1 to obtain target compound REX-6 in yield: 33 percent. MSm/z [ ESI ]]：518.1[M+1]。1H-NMR(400MHz，DMSO6)：＝8.660-8.639(d，J＝8.4Hz，1H)，8.445(s，1H)，8.376(s，1H)，7.861-7.841(d，J＝8.0Hz，1H)，7.750(m，1H)，7.436(s，1H)，7.390(m，1H),6.975(s，1H)，4.708(m，1H)，4.041(m，1H)，3.503(m，3H)，3.073(m，2H)，2.607(m，2H)，2.070(s，3H)，1.997(m，2H)，1.711(m，4H)，1.180(m，6H)。
Example preparation of 72- (4- (4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -2-1(2H) pyridonyl) -1-piperidine) acetamide [ number REX-12 ]
The synthetic route is as follows:
Figure BDA0000701187500000212
Figure BDA0000701187500000221
synthesis scheme 1: synthesis of intermediate 2- (4- (4-amino-2-oxopyridin-1 (2h) yl) piperidinyl) acetamide (i.e. Compound 1-3)
Step 1: preparation of intermediate 4-methanesulfonyloxypiperidine-1-acetamide (i.e., Compound 1-2)
Dissolving raw materials of N-acetamide-4-hydroxypiperidine (namely a compound 1-1, 10.0g and 63.2mmol) and triethylamine (17.5mL and 127mmol) in dichloromethane (100mL), slowly adding dropwise methylsulfonyl chloride (5.1mL and 63.2mmol) at the temperature of below 5 ℃, and reacting for 30 minutes at the temperature of 5-15 ℃. After the reaction was completed, the temperature was returned to room temperature, and the mixture was diluted with 200mL of water and extracted with methylene chloride. The organic phase was washed twice with sodium bicarbonate, dried over anhydrous sodium sulfate, and concentrated to give compound 1-2(8.0g), yield: 53.5 percent. MSm/z [ ESI ]: 237.3[ M +1 ].
Step 2: preparation of intermediate 2- (4- (4-amino-2-oxopyridin-1 (2H) yl) piperidinyl) acetamide (Compound 1-3)
4-amino-2-hydroxypyridine (3.0g,27.9mmol) was dissolved in DMF (50mL) in an ice-water bath, and sodium hydride (1.2g,30.8mmol) was slowly added to the reaction system and stirred at 0 ℃ for 15 minutes. Then, compound 1-2(6.6g,28.0mmol) was added to the reaction system, and the mixture was heated to 45 ℃ and stirred overnight. After cooling, the mixture was poured into 400mL of water, extracted with ethyl acetate, dried, concentrated and subjected to silica gel column chromatography to obtain compound 1-3(3.6g) in yield: 51.6 percent. MSm/z [ ESI ]: 294.4[ 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)
Raw materials of 2-fluoronitrobenzene (namely, compound 2-1, 10g,70mmol) and isopropyl mercaptan (5.4g, 7mmol)0mmol) and potassium carbonate (25g,177mmol) were added to dry DMF (100mL) under nitrogen and stirred overnight at 100-110 ℃. The reaction mixture was cooled to room temperature, water (200mL) was added, and extraction was performed with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to give compound 2-2(10g), yield: 72 percent. MSm/z [ ESI ]]：198.2[M+1]。1H-NMR(400MHz，CDCl3)：＝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,65.97mmol) in dichloromethane (100mL) was added m-CPBA (25.67g,149.42mmol) in portions with stirring at 0 ℃ and, after the addition, reacted at 0 ℃ for 16 hours. The reaction mixture was washed with saturated sodium bicarbonate solution, dried, concentrated and subjected to silica gel column chromatography to obtain compound 2-3(10.66g) in yield: 70 percent. MSm/z [ ESI ]]：230.2[M+1]。1H-NMR(400MHz，CDCl3)：＝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)
Compound 2-3(20g,87.3mmol) and 10% palladium on charcoal (2.0g) were added to dry methanol (250mL), displaced with hydrogen, and reacted at 60 ℃ for 2 hours. After cooling, the mixture was separated by filtration through celite, and dried and concentrated over anhydrous sodium sulfate to obtain compound 2-4(17.3g) in yield: 95 percent. MSm/z [ ESI ]]：200.2[M+1]。1H-NMR(400MHz，DMSO6)：＝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 ice-water bath conditions, compounds 2-4(30g,150mmol) were dissolved in DMF (300mL), and sodium hydride (7.23g,300mmol) was slowly added to the reaction system and stirred at 0 ℃ for 15 minutes. 2,4, 5-trichloropyrimidine (33.1g,180mmol) was added dropwise to the reaction system, and the reaction was stirred at room temperature overnight. 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 compound 2-5(17.3g) in yield: 32 percent. MSm/z [ ESI ]: 370.2[ M +1 ]. 1H-NMR (400MHz, DMSO 6): 9.822(s, 1H), 7.343-7.323(d, J-8.0 Hz, 1H), 7.911-7.839(m, 2H), 3.585-3.484(m, 1H), 1.176-1.159(d, J-6.8 Hz, 6H).
Synthesis scheme 3: synthesis of target compound 2- (4- (4- ((5-chloro-4- ((2- (isopropylsulfonyl) phenyl) amine) pyrimidin-2-amino) -2-1(2H) pyridonyl) -1-piperidine) acetamide (i.e. REX-12)
Step 1: compound 1 to 3(0.9g,3.6mmol), compound 2 to 5(1.4g,3.6mmol), cesium carbonate (3.5g,10.7mmol), catalyst 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (0.45g,0.75mmol) and palladium acetate (81mg,0.36mmol) were dissolved in dioxane (20mL) and charged into a sealed tube, and after replacing the air with nitrogen, the reaction was carried out at 95 ℃ for 18 hours. Spin-drying the solvent, adding ethyl acetate and water, extracting, drying the organic phase, and purifying by silica gel column chromatography to obtain the target compound REX-12(0.75g), yield: 37 percent. MSm/z [ ESI ]: 561.1[ M +1 ].
Example 8 measurement of ALK kinase inhibitory Activity and binding Rate to related mutation sites
Selecting the compounds REX-1 to REX-4 prepared in the embodiments 1 to 4 of the invention, and adopting
Figure BDA0000701187500000241
(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 BDA0000701187500000242
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. LanthaScreen Eu kinase binding assay by adding Eu tagAntibodies or anti-tag antibodies detect Alexa Fluor conjugate or kinase "tracer" binding. 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 also has better binding activity on 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-1 <50 <50
REX-2 <50 <50
REX-3 <50 <50
REX-4 <50 <50
Example 9 cell proliferation assay
Test compounds: compounds REX-1 and REX-2 prepared in examples 1-2 of the present invention.
Cell lines: lung cancer cell line NCI-H2228, purchased from bai biotechnology limited of beijing.
The method comprises the following steps: the cell line NCI-H2228 was cultured in 20% FBS (fetal bovine serum) (Gibco) +1640+ 1% double antibody. Then, the NCI-H2228 cells with good growth state are taken, 5000 cells/well are respectively inoculated on a 96-well cell plate, the cell plate is placed at the temperature of 37 ℃ and contains 5 percent 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 BDA0000701187500000261
as a result: see table 2.
TABLE II example cell proliferation IC of Compounds on Lung cancer cell lines50Value of
IC50(μM) NCI-H2228
REX-1 <1
REX-2 <10

Claims (10)

1. A pyridone protein kinase inhibitor is a compound with the following structural formula and pharmaceutically acceptable salts thereof:
Figure FDA0002573935990000011
wherein R is1Selected from hydrogen,
Figure FDA0002573935990000012
R0、R2、R3、R4、R11Each independently selected from hydrogen, halogen, C1-6Alkyl, halo C1-6Alkyl radical, C1-6Alkoxy, halo C1-6Alkoxy radical, C1-6Cycloalkyl, halo C1-6Cycloalkyl, cyano or amino;
R6selected from hydrogen, C1-6Alkyl or
Figure FDA0002573935990000013
R10Selected from hydrogen, C1-6Alkyl or a 3-6 membered heterocyclic ring selected from the group consisting of N, O heteroatoms;
and m, p are selected from any value of 0, 1, 2, 3, 4 and 5, and n is selected from any value of 0, 1, 2, 3, 4,5 and 6.
2. The pyridone protein kinase inhibitor according to claim 1, wherein: the 3-6 membered heterocyclic group with N, O heteroatoms is one of pyridyl, piperidyl, pyrimidyl, furyl and morpholinyl.
3. The pyridone protein kinase inhibitor according to claim 1, wherein: the halogen is one of fluorine, chlorine, bromine and iodine.
4. A pyridone protein kinase inhibitor selected from the following compounds:
Figure FDA0002573935990000014
Figure FDA0002573935990000021
5. a pharmaceutical composition comprising a compound as defined in claim 1 or 4, or a pharmaceutically acceptable salt thereof, as an active ingredient, together with one or more pharmaceutically acceptable carriers.
6. Use of a compound as defined in claim 1 or 4, 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.
7. Use of a compound as defined in claim 1 or 4, 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.
8. Use according to claim 7, characterized in that: the disease is selected from cell proliferative diseases.
9. Use according to claim 8, characterized in that: the cell proliferative disease is a tumor.
10. Use according to claim 9, characterized in that: the cell proliferative disease is selected from non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, 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.
CN201510183879.2A 2015-04-17 2015-04-17 Pyridone protein kinase inhibitors Active CN106146468B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510183879.2A CN106146468B (en) 2015-04-17 2015-04-17 Pyridone protein kinase inhibitors

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510183879.2A CN106146468B (en) 2015-04-17 2015-04-17 Pyridone protein kinase inhibitors

Publications (2)

Publication Number Publication Date
CN106146468A CN106146468A (en) 2016-11-23
CN106146468B true CN106146468B (en) 2020-12-01

Family

ID=58057670

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510183879.2A Active CN106146468B (en) 2015-04-17 2015-04-17 Pyridone protein kinase inhibitors

Country Status (1)

Country Link
CN (1) CN106146468B (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109422733A (en) * 2017-09-03 2019-03-05 上海美志医药科技有限公司 One kind inhibits and the compound for the tyrosine protein kinase ALK that degrades
CN111170996B (en) * 2018-11-09 2021-12-28 天津大学 Pyrimidine derivative with ALK inhibitory activity and synthetic method and application thereof

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005080350A1 (en) * 2004-02-23 2005-09-01 Glaxo Group Limited Pyrimidine derivatives as cannabinoid receptor modulators
WO2008118822A1 (en) * 2007-03-23 2008-10-02 Rigel Pharmaceuticals, Inc. Compositions and methods for inhibition of the jak pathway
CN101616895A (en) * 2006-12-08 2009-12-30 Irm责任有限公司 Compound and composition as kinases inhibitor
WO2010142766A2 (en) * 2009-06-10 2010-12-16 Cellzome Limited Pyrimidine derivatives as zap-70 inhibitors
CN101921236A (en) * 2003-03-14 2010-12-22 诺瓦提斯公司 Can be used for treating 2 of neoplastic disease, inflammatory and disorder of immune system, 4-two (phenylamino) pyrimidine
CN102083800A (en) * 2008-06-27 2011-06-01 阿维拉制药公司 Heteroaryl compounds and uses thereof
CN102105150A (en) * 2008-05-21 2011-06-22 阿里亚德医药股份有限公司 Phosphorous derivatives as kinase inhibitors
CN102421762A (en) * 2009-03-18 2012-04-18 贝林格尔.英格海姆国际有限公司 Substituted pyrimidines for the treatment of cancer

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101921236A (en) * 2003-03-14 2010-12-22 诺瓦提斯公司 Can be used for treating 2 of neoplastic disease, inflammatory and disorder of immune system, 4-two (phenylamino) pyrimidine
WO2005080350A1 (en) * 2004-02-23 2005-09-01 Glaxo Group Limited Pyrimidine derivatives as cannabinoid receptor modulators
CN101616895A (en) * 2006-12-08 2009-12-30 Irm责任有限公司 Compound and composition as kinases inhibitor
WO2008118822A1 (en) * 2007-03-23 2008-10-02 Rigel Pharmaceuticals, Inc. Compositions and methods for inhibition of the jak pathway
CN102105150A (en) * 2008-05-21 2011-06-22 阿里亚德医药股份有限公司 Phosphorous derivatives as kinase inhibitors
CN102083800A (en) * 2008-06-27 2011-06-01 阿维拉制药公司 Heteroaryl compounds and uses thereof
CN102421762A (en) * 2009-03-18 2012-04-18 贝林格尔.英格海姆国际有限公司 Substituted pyrimidines for the treatment of cancer
WO2010142766A2 (en) * 2009-06-10 2010-12-16 Cellzome Limited Pyrimidine derivatives as zap-70 inhibitors

Also Published As

Publication number Publication date
CN106146468A (en) 2016-11-23

Similar Documents

Publication Publication Date Title
JP6790040B2 (en) Novel compounds and compositions for inhibiting FASN
AU2013230146B2 (en) 2-amino, 6-phenyl substituted pyrido [2, 3 - d] pyrimidine derivatives useful as Raf kinase inhibitors
ES2671354T3 (en) DNA-PK inhibitors
TWI527800B (en) 1-(arylmethyl)quinazoline-2,4(1h,3h)-diones as parp inhibitors and the use thereof
TW202043212A (en) Shp2 inhibitor and application thereof
JP7337951B2 (en) Nitrogen-containing aromatic heterocyclic amide derivatives for treating cancer
KR102359993B1 (en) Pyrimido [5,4-b] indolizine or pyrimido [5,4-b] pyrrolizine compound, preparation method and use thereof
CN114057771B (en) Macrocyclic compounds, their preparation and use
KR20090090365A (en) Pyridine or pyrimidine derivative having excellent cell growth inhibition effect and excellent anti-tumor effect on cell strain having amplification of hgfr gene
JP7320823B2 (en) Use of pteridinone derivatives as EGFR inhibitors
CN112724145A (en) Pyrazine derivatives for inhibiting SHP2 activity
WO2017114383A1 (en) Novel kinase inhibitor against wild-type egfr and mutated egfr
EA019723B1 (en) cMET INHIBITORS
KR20100043288A (en) 5-(4-(haloalkoxy)phenyl)pyrimidine-2-amine compounds and compositions as kinase inhibitors
CN110483485A (en) Pyrimidines, preparation method and medical usage
CN106536510B (en) The 2-aminopyridine albuminoid kinase inhibitor that pyridine replaces
WO2021175271A1 (en) Novel hpk1 inhibitor, and preparation method therefor and application thereof
CN110546145B (en) Azaaryl derivative, preparation method and pharmaceutical application thereof
WO2020125759A1 (en) Compound as wnt signal pathway inhibitor and medical use thereof
CN106336382B (en) 4-saturated cyclic substituted aniline protein kinase inhibitor
WO2018157737A1 (en) Multi-target kinase inhibitor
CN106146468B (en) Pyridone protein kinase inhibitors
CN109020957B (en) Heterocyclic compounds as MNK inhibitors
CN103319404A (en) Nitroxoline derivatives and use of the nitroxoline derivatives as angiogenesis inhibitors
WO2020052489A1 (en) Preparation for 6-amino-1h-pyrazolo[3,4-d]pyrimidine-based jak kinase inhibitor and application thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
TA01 Transfer of patent application right
TA01 Transfer of patent application right

Effective date of registration: 20200123

Address after: 310011 No. 108 Tong Road, Hangzhou, Zhejiang, Yuhang

Applicant after: Wu Yaodong

Address before: Hangzhou City, Zhejiang province Xiaoshan District 311202 North Street Jincheng Road No. 1038, room 817

Applicant before: Hangzhou REX Pharmaceutical Co.,Ltd.

TA01 Transfer of patent application right
TA01 Transfer of patent application right

Effective date of registration: 20201027

Address after: 817, room 1038, Jincheng Road, Xiaoshan District, Zhejiang, Hangzhou 311202, China

Applicant after: Hangzhou REX Pharmaceutical Co.,Ltd.

Address before: 310011 No. 108 Tong Road, Hangzhou, Zhejiang, Yuhang

Applicant before: Wu Yaodong

GR01 Patent grant
GR01 Patent grant