Dihydropteridinone derivatives containing phenyl biheteroaryl groups and uses thereof

Abstract

The present invention relates to dihydropteridone derivatives containing phenyl biheteroaryl groups represented by general formula I and their optical isomers, pharmaceutically acceptable salts, solvates or prodrugs, their preparation methods and pharmaceutical compositions containing compounds represented by general formula I as active ingredients, wherein substituents R1, R2, R3, R4, R5, X, m and A have the meanings given in the specification. The present invention also relates to the use of compounds represented by general formula I and their pharmaceutically acceptable salts, solvates or prodrugs in the preparation of drugs for treating diseases caused by abnormally high expression of PLK1 kinase, in particular, in the preparation of drugs for treating and / or preventing cancer.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, in particular to a dihydropteridone derivative containing a phenyl biheteroaryl group and its use in preparing a drug for treating and / or preventing cancer. Background Art

[0002] Cancer, also known as malignant tumor, is a malignant disease caused by abnormal differentiation and proliferation of normal tissue cells. Cancer is the second leading cause of death in the world and has gradually become an increasingly serious public health problem. According to statistics, there were 18.1 million cancer cases and 9.6 million cancer deaths in 2018. The current form of cancer in my country is serious. According to statistics, of the 9.6 million cancer deaths in the world in 2018, 30% (2.9 million) of the deaths occurred in China. The cancer incidence rate in China (201.7 / 100,000) is comparable to the global total incidence rate (197.9 / 100,000), but the cancer mortality rate in China is much higher (130.1 / 100,000 in China, 102.6 / 100,000 in the UK, and 91.0 / 100,000 in the United States). Cancer has become the leading cause of death among Chinese residents and has placed a heavy burden on a large number of families. Therefore, the research on anti-tumor drugs and their related target inhibitors has important clinical and social value.

[0003] PLKs (Polo-like kinases) are a class of Ser / Thr protein kinases found in eukaryotes. They have a unique cell cycle regulation mechanism and play an important role in tumor cell proliferation and differentiation. Using PLKs as a target to find anti-tumor drugs with a new mechanism has become an important branch of targeted drug research.

[0004] At present, among all the members of the PLKs family, researchers have conducted the most thorough research on PLK1. The structure of PLK1 in the human body includes two important domains, namely the N-terminal kinase domain, which contains 252 amino acids; and the C-terminal PDB domain, which contains 60 to 70 amino acids. The crystal structure analysis of the two important domains of the protein, the C-terminal and the N-terminal, has been completed and reported in recent years. The crystal structure of the N-terminal kinase domain of PLK1 is a classic protein kinase structure, that is, the N-terminal antiparallel β-fold protrusion and the C-terminal α-helical protrusion form a pocket for ATP binding in space, and the highly conserved hinge region connecting the two protrusions is the main site of the main chain and ligand binding reaction. Compared with other traditional kinase structures, some specific sites also exist in the binding region of PLK1 kinase and inhibitors: such as phenylalanine at the bottom of the ATP binding pocket, cysteine ​​at the top (leucine in common kinases), the pocket in the hinge region, and the positively charged residues outside the adenine pocket. These specific sites provide a structural basis for the design of specific PLK1 inhibitors.

[0005] The importance of PLK in key cell cycle processes is obvious. Its most conserved function is to precisely control the microtubules in the centrosome, spindle, and kinetochore. The expression of the PLK1 gene is regulated by the cell cycle and its expression level changes with the stage of the cell cycle. Usually in the early stage of mitosis, PLK1 accumulates in the center of the spindle, and gradually migrates from the spindle to the equatorial plate after the middle and late stages of mitosis. At the end of mitosis, PLK1 accumulates in the central region of the spindle and phosphorylates HsCYK4 to activate the RhoA enzyme. The RhoA enzyme completes a cell cycle by regulating the contraction of the contractile ring and then the cytoplasm divides. Therefore, PLK1 expression is low in the G1 and S phases, gradually increases in the G2 phase, and reaches a peak in the M phase. After cell division is completed, PLK1 expression will drop sharply and then enter the next cell cycle. In addition to regulating the progress of mitosis, PLK1 also plays an important role in the maturation of the centrosome, the maturation of sister chromosomes, the regulation of the microtubule organizing center, and cell division.

[0006] At present, with the continuous development of medicinal chemistry, oncology and pharmacology, the global research on PLK1 targeted drugs has achieved certain results. At present, there are many PLK1 inhibitors in clinical research around the world, including dihydropteridinone compounds BI2536 (Phase II) and Volasertib (Phase III), tricyclic compounds NMS-P397 (Phase II), thiophene compounds GSK461364 (Phase I), pyrimidine and diazepine ketone compounds TAK960 (Phase I) and other Rigosertib (Phase III). However, due to the high homology between PLK subtypes and the problems caused by the administration method, no PLK1 small molecule inhibitor has been approved for marketing so far. There is an urgent need to find more new and efficient PLK1 inhibitors in clinic to provide new options for the treatment of diseases such as tumors.

[0007]

[0008] Structures of PLK1 inhibitors in clinical trials. Summary of the invention

[0009] The present invention aims to provide a dihydropteridone derivative containing a phenyl biheteroaryl group and its use in preparing a drug for treating and / or preventing cancer.

[0010] To achieve the above purpose, the present invention adopts the following technical solution:

[0011] A dihydropteridone derivative containing a phenylheteroaryl group, wherein the derivative is a compound represented by general formula I and a salt, solvate or prodrug thereof,

[0012]

[0013] in,

[0014] R1 is (C1-C6) alkyl, (C3-C6) cycloalkyl;

[0015] R2 is (C1-C6) alkyl, (C3-C6) cycloalkyl, 3-6 membered heterocyclic group, (C6-C 12 )spirocyclic group, 6-12 membered heterospirocyclic group, (C6-C 10 ) aryl, 5-10 membered heteroaryl, the alkyl, cycloalkyl, heterocyclyl, spirocyclyl, heterospirocyclyl, aryl or heteroaryl is optionally substituted by 1-3 identical or different R6;

[0016] R6 is hydrogen, halogen, hydroxy, cyano, amino, nitro, (C1-C6) alkyl, (C1-C6) haloalkyl, (C1-C6) alkoxy;

[0017] R3 is hydrogen, (C1-C6)alkoxy, (C3-C6)cycloalkoxy which is unsubstituted or substituted by at least one of the following groups; the following groups are halogen, hydroxyl or amino;

[0018] Ring A is selected from (C6-C 10 ) aryl, 5-6 membered heteroaryl, said aryl or heteroaryl being optionally substituted by 1-3 identical or different R9;

[0019] R9 is hydrogen, halogen, hydroxy, cyano, amino, nitro, (C1-C6)haloalkyl, (C1-C6)alkyl, (C1-C6)alkoxy;

[0020] R4 and R5 are the same or different and are independently selected from hydrogen, (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl, (C2-C6) alkenyl, (C2-C6) alkynyl, dimethylaminoethyl, and the alkyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 identical or different R7;

[0021] or R4 and R5 together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclic group containing at least one heteroatom; the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, the heterocyclic group is optionally substituted with one or more oxo groups, or the heterocyclic group is optionally substituted with 0-3 identical or different R8 groups;

[0022] R7 and R8 are the same or different and are independently selected from hydrogen, hydroxy, halogen, amino substituted with 0-2 (C1-C6) alkyl groups, cyano, (C1-C6) alkyl acyl, (C1-C6) alkyl sulfonyl, (C1-C6) alkyl which is unsubstituted or substituted with at least one of the following groups, (C1-C6) alkoxy, (C3-C6) cycloalkyl, (C3-C6) alkylsulfonyl, 10 )cycloalkyl-(C1-C4)alkyl; the following groups are hydroxy, amino or halogen;

[0023] X is an oxo-substituted or unoxo-substituted (C1-C6) alkyl group;

[0024] m is an integer from 0 to 1;

[0025] The above heterocyclic group or heteroaryl group may contain 1 to 4 heteroatoms, which may be the same or different and are selected from N, O and S.

[0026] Preferably, in the general formula I,

[0027] R1 is (C1-C4) alkyl, (C3-C5) cycloalkyl;

[0028] R2 is (C1-C6) alkyl, (C3-C6) cycloalkyl, 3-6 membered heterocyclic group, (C6-C10 ) aryl, 5-6 membered heteroaryl, the alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl group is optionally substituted by 1-3 identical or different R6;

[0029] R6 is hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethyl, (C1-C6) alkyl, (C1-C6) alkoxy;

[0030] R3 is unsubstituted or substituted by at least one of the following groups (C1-C3) alkoxy; the following group is halogen, hydroxyl or amino;

[0031] Ring A is selected from phenyl, 5-6 membered heteroaryl, and the phenyl or heteroaryl is optionally substituted by 1-3 identical or different R9;

[0032] R9 is hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethyl, (C1-C3) alkyl, (C1-C3) alkoxy;

[0033] R4 and R5 are the same or different and are independently selected from hydrogen, (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl, dimethylaminoethyl, wherein the alkyl, cycloalkyl, heterocycloalkyl are optionally substituted by 1-3 identical or different R7;

[0034] or R4 and R5 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing at least one heteroatom; the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, the heterocyclic group is optionally substituted with one or more oxo groups, or the heterocyclic group is optionally substituted with 0-3 identical or different R8 groups;

[0035] R7 and R8 are the same or different and are independently selected from hydrogen, hydroxyl, halogen, amino substituted with 0-2 (C1-C4) alkyl groups, cyano, (C1-C4) alkylacyl, (C1-C4) alkylsulfonyl, (C1-C4) alkyl which is unsubstituted or substituted with at least one of the following groups, (C1-C4) alkoxy, (C3-C6) cycloalkyl, (C3-C6) cycloalkyl-(C1-C4) alkyl; the following groups are hydroxyl, amino or halogen;

[0036] X is an oxo-substituted or unoxo-substituted (C1-C4) alkyl group;

[0037] m is an integer from 0 to 1;

[0038] The above heterocyclic group or heteroaryl group may contain 1 to 4 heteroatoms, which may be the same or different and are selected from N, O and S.

[0039] Preferably, in the general formula I,

[0040] R1 is (C1-C4) alkyl;

[0041] R2 is (C1-C6) alkyl, (C3-C6) cycloalkyl, 3-6 membered heterocyclic group, (C6-C 10 ) aryl, 5-6 membered heteroaryl, the alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl group is optionally substituted by 1-3 identical or different R6;

[0042] R6 is hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethyl, (C1-C6) alkyl, (C1-C6) alkoxy;

[0043] R3 is (C1-C3) alkoxy;

[0044] Ring A is selected from

[0045] R4 and R5 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl, (C3-C5) cycloalkyl, (C3-C6) heterocycloalkyl, and the alkyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 identical or different R7;

[0046] or R4 and R5 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing at least one heteroatom; the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, the heterocyclic group is optionally substituted with one or more oxo groups, or the heterocyclic group is optionally substituted with 0-3 identical or different R8 groups;

[0047] R7 and R8 are the same or different and are independently selected from hydrogen, hydroxyl, halogen, amino substituted with 0-2 (C1-C6) alkyl groups, cyano, (C1-C3) alkylacyl, (C1-C3) alkylsulfonyl, (C1-C3) alkyl unsubstituted or substituted with at least one of the following groups, (C1-C3) alkoxy, (C3-C6) cycloalkyl, (C3-C6) cycloalkyl-(C1-C4) alkyl; the following groups are hydroxyl, amino or halogen;

[0048] X is an oxo-substituted or unoxo-substituted (C1-C4) alkyl group;

[0049] m is an integer from 0 to 1;

[0050] The above heterocyclic group or heteroaryl group may contain 1 to 4 heteroatoms, which may be the same or different and are selected from N, O and S.

[0051] More preferably, in the general formula I,

[0052] R1 is methyl

[0053] R2 is (C3-C6) alkyl, (C3-C6) cycloalkyl, 3-6 membered heterocyclyl, phenyl, thienyl, furyl, naphthyl, wherein the alkyl, cycloalkyl, heterocyclyl, phenyl, thienyl, furyl, naphthyl is optionally substituted by 1-3 identical or different R6;

[0054] R6 is hydrogen, halogen, (C1-C4) alkyl, (C1-C4) alkoxy, trifluoromethyl, hydroxyl, cyano;

[0055] R3 is (C1-C3) alkoxy;

[0056] Ring A is selected from

[0057] R4 and R5 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl, (C3-C5) cycloalkyl, (C3-C6) heterocycloalkyl, and the alkyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 identical or different R7;

[0058] or R4 and R5 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing at least one heteroatom; the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, the heterocyclic group is optionally substituted with one or more oxo groups, or the heterocyclic group is optionally substituted with 0-3 identical or different R8 groups;

[0059] R7 and R8 are the same or different and are independently selected from hydrogen, hydroxyl, halogen, amino substituted with 0-2 (C1-C6) alkyl groups, cyano, (C1-C3) alkylacyl, (C1-C3) alkylsulfonyl, (C1-C3) alkyl unsubstituted or substituted with at least one of the following groups, (C1-C3) alkoxy, (C3-C6) cycloalkyl, (C3-C6) cycloalkyl-(C1-C4) alkyl; the following groups are hydroxyl, amino or halogen;

[0060] X is an oxo-substituted or unoxo-substituted (C1-C4) alkyl group;

[0061] m is an integer from 0 to 1;

[0062] The above heterocyclic group or heteroaryl group may contain 1 to 4 heteroatoms, which may be the same or different and are selected from N, O and S.

[0063] Still more preferably, in the general formula I,

[0064] R1 is methyl

[0065] R2 is (C3-C6) alkyl, (C3-C6) cycloalkyl, 3-6 membered heterocyclyl, phenyl, thienyl, furyl, naphthyl, wherein the alkyl, cycloalkyl, heterocyclyl, phenyl, thienyl, furyl, naphthyl is optionally substituted by 1-3 identical or different R6;

[0066] R6 is hydrogen, halogen, (C1-C4) alkyl, (C1-C4) alkoxy, trifluoromethyl, hydroxyl, cyano;

[0067] R3 is methoxy;

[0068] Ring A is selected from

[0069] R4 and R5 are the same or different and are independently selected from hydrogen, (C1-C6) alkyl, (C3-C5) cycloalkyl, (C3-C6) heterocycloalkyl, and the alkyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 identical or different R7;

[0070] or R4 and R5 together with the nitrogen atom to which they are attached form

[0071] X is CH2 or CO;

[0072] m is an integer from 0 to 1;

[0073] The above heterocyclic group or heteroaryl group may contain 1 to 4 heteroatoms, which may be the same or different and are selected from N, O and S.

[0074] More preferably, the derivative is a compound of formula I and its stereoisomers, pharmaceutically acceptable salts, solvates or prodrugs,

[0075] (R)-8-Cyclopentyl-2-((4-(4-((dimethylamino)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0076] (R)-8-Cyclopentyl-2-((4-(4-((4-cyclopentylpiperazin-1-yl)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0077] (R)-8-Cyclopentyl-2-((4-(4-((diethylamino)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0078] (R)-8-Cyclopentyl-7-ethyl-2-((4-(4-((4-ethylpiperazin-1-yl)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0079] (R)-8-Cyclopentyl-2-((4-(4-((cyclopropylamino)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0080] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(piperazin-1-ylmethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0081] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((4-methylpiperazin-1-yl)methyl)-1,2,4-oxadiazol-3-yl))phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0082] (R)-8-Cyclopentyl-2-((4-(5-((4-cyclopentylpiperazin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0083] (R)-8-Cyclopentyl-7-ethyl-2-((4-(5-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-1,2,4-oxadiazol-3-yl))-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0084] (R)-8-Cyclopentyl-7-ethyl-2-((4-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0085] (R)-8-Cyclopentyl-2-((4-(5-(((3S,5R)-3,5-dimethylpiperazin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0086] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(4-((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0087] (R)-8-Cyclopentyl-7-ethyl-2-((4-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0088] (R)-8-Cyclopentyl-2-((4-(4-(((3S,5R)-3,5-dimethylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0089] (R)-8-Cyclopentyl-7-ethyl-2-((4-(4-((4-isopropylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0090] (R)-8-Cyclopentyl-2-((4-(5-((dimethylamino)methyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0091] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((3-methylpiperazin-1-yl)methyl)-1,3,4-oxadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0092] (R)-8-Cyclopentyl-2-((4-(5-(((2-(dimethylamino)ethyl)(methyl)amino)methyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0093] (R)-8-Cyclopentyl-7-ethyl-2-((4-(5-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-1,3,4-oxadiazol-2-yl))-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0094] (R)-8-Cyclopentyl-2-((4-(5-((3-(dimethylamino)pyrrolidin-1-yl)methyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0095] (R)-8-Cyclopentyl-7-ethyl-2-((4-(5-(((R)-2-(hydroxymethyl)pyrrolidin-1-yl)methyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0096] (R)-8-Cyclohexyl-7-ethyl-2-((4-(5-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-1,3,4-oxadiazol-2-yl))-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0097] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((3-methylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0098] (R)-8-Cyclopentyl-2-((4-(5-(((3S,5R)-3,5-dimethylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0099] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((4-methylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl))phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0100] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(piperazin-1-ylmethyl)-1,3,4-thiadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0101] (R)-2-((4-(5-((4-acetylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl)-2-methoxyphenyl)amino)-8-cyclopentyl-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one.

[0102] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(piperazin-1-ylmethyl)oxazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0103] (R)-8-Cyclopentyl-2-((4-(2-((dimethylamino)methyl)-1H-pyrrol-1-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0104] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((4-methylpiperazin-1-yl)methyl)isoxazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropyridin-6(5H)-one

[0105] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(3-(4-methylpiperazine-1-carbonyl)-1,2,4-oxadiazol-5-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0106] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)-1,3,4-oxadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0107] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)-1,3,4-thiadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0108] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(4-(4-methylpiperazine-1-carbonyl)-1H-1,2,3-triazol-1-yl)phenyl))amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0109] (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)isoxazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropyridin-6(5H)-one

[0110] The compound salt is a salt formed with an acid, and the acid is selected from: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, trifluoroacetic acid or aspartic acid.

[0111] A pharmaceutical composition comprising a compound of general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0112] A use of a compound, wherein the compound of general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof are used in the preparation of a drug for preventing or treating a disease associated with a PLK1 kinase inhibitor.

[0113] Moreover, according to some common methods in the field to which the present invention belongs, the dihydropteridinone derivatives of the general formula I in the present invention can be reacted with acids to form pharmaceutically acceptable salts. Pharmaceutically acceptable addition salts include inorganic acid and organic acid addition salts, and the salts with the following acids are particularly preferred: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalene disulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, etc.

[0114] In addition, the present invention also includes prodrugs of the derivatives of the present invention. Prodrugs of the derivatives of the present invention are dihydropteridone derivatives of general formula I, which may have weak activity or even no activity themselves, but after administration, they are converted into the corresponding biologically active form under physiological conditions (e.g., by metabolism, solvolysis or other means).

[0115] In the above substituents, "halogen" refers to fluorine, chlorine or bromine; "alkyl" refers to straight-chain or branched alkyl; "cycloalkyl" refers to substituted or unsubstituted cycloalkyl; "alkoxy" refers to straight-chain or branched alkoxy; "alkenyl" refers to straight-chain or branched alkenyl; "alkynyl" refers to straight-chain or branched alkynyl; "aryl" refers to phenyl or naphthyl with or without substituents; "heteroaryl" refers to a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, and S, and the ring system is aromatic.

[0116] We have found that the compounds of the present invention have the activity of inhibiting tumor cell growth in vitro. Therefore, they can be used to prepare drugs for treating and / or preventing cancer, such as cancer of the breast, lung, liver, kidney, colon, rectum, stomach, prostate, bladder, uterus, pancreas, bone marrow, testis, ovary, lymph, soft tissue, head and neck, thyroid, esophagus and leukemia, neuroblastoma, etc.

[0117] Through in vitro inhibition of human breast cancer cells MDA-MB-231 and human myelomonocytic leukemia cells MV-4-11, the compounds of the present invention have significant inhibitory effects on breast cancer cells and leukemia cells. Through the test of PLK1 protein activity, it was found that the compounds of the present invention have a good inhibitory effect on PLK1 protein activity, and are particularly useful for preparing drugs for treating and / or preventing breast cancer and leukemia.

[0118] A pharmaceutical composition comprising a compound represented by general formula I and its stereoisomers, pharmaceutically acceptable salts, solvates or prodrugs mixed with a pharmaceutically acceptable carrier.

[0119] A use of a compound and a pharmaceutical composition, wherein the compound of the general formula I and its stereoisomers, pharmaceutically acceptable salts, solvates or prodrugs or a composition containing the compound of the general formula I and its stereoisomers, pharmaceutically acceptable salts, solvates or prodrugs are used in the preparation of a drug for treating and / or preventing PLK1-mediated diseases.

[0120] The preparation for treating and / or preventing PLK1-mediated diseases includes but is not limited to breast cancer, glioblastoma, lung cancer, head and neck cancer, pancreatic cancer, gastric cancer, and brain cancer, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, biliary tract cancer, bladder cancer, colorectal cancer, osteosarcoma, ovarian cancer, multiple myeloma, skin cancer, melanoma, leukemia or kidney cancer.

[0121] The present invention also includes prodrugs of the derivatives of the present invention. Prodrugs of the derivatives of the present invention are derivatives of general formula (I) which may themselves have weak activity or even no activity, but after administration, are converted into the corresponding biologically active form under physiological conditions (e.g., by metabolism, solvolysis or other means).

[0122] The present invention also includes a pharmaceutical composition comprising a compound of formula I and their pharmaceutically acceptable salts and / or solvates as an active ingredient and a pharmaceutically acceptable carrier; the compounds of the present invention may also be used in combination with other active ingredients as long as they do not produce other adverse effects, such as allergic reactions.

[0123] The carriers used in the pharmaceutical composition of the present invention are common types available in the pharmaceutical field, including: binders, lubricants, disintegrants, cosolvents, diluents, stabilizers, suspending agents, pigments, flavoring agents, etc. for oral preparations; pH regulators, osmotic pressure regulators, solubilizers, stabilizers, etc. for injectable preparations; bases, diluents, lubricants, preservatives, etc. for local preparations. Pharmaceutical preparations can be administered orally, parenterally (e.g., intravenously, subcutaneously, intraperitoneally, etc.) or topically (e.g., ocular, nasal, sublingual, skin, etc.), and if some drugs are unstable under gastric conditions, they can be formulated into enteric-coated tablets.

[0124] The exact amount of the compound of the present invention for treating cancer, especially diseases caused by abnormal PLK1, will vary from subject to subject, depending on the type, age and general condition of the subject, the severity of the disease being treated, the specific compound used and the mode of administration, such as the route and frequency of administration, etc. An ordinary technician in this field can determine the appropriate effective amount using only routine experimental methods.

[0125] The compound can be administered in an amount of about 0.1 to 160 mg / kg body weight per day, preferably 1 to 60 mg / kg body weight per day. It is understood that the dosage may vary depending on the patient's needs, the disease caused by the abnormal PLK1 being treated, and the specific compound used. Moreover, it is understood that the initial dose administered may be increased beyond the upper limit in order to quickly reach the desired blood level, or the initial dose may be less than the optimal value, and the daily dose may be gradually increased during treatment, depending on the specific situation. If necessary, the daily dose may also be divided into multiple doses, for example 2-4 times a day.

[0126] Mammal means a human or an animal.

[0127] The amount of active ingredient, i.e. the compound according to the invention, in the pharmaceutical composition and its unit dosage form may vary, depending on the specific application, the efficacy of the specific compound and the desired concentration. Generally speaking, the content of the active ingredient will be between 0.5% and 90%, based on the total weight of the composition.

[0128] In combination therapy, the compound of the present invention and other compounds may be administered simultaneously or separately. When administered simultaneously, the compound of the present invention and other compounds may be combined in a single pharmaceutical composition or in separate compositions.

[0129] The examples and preparations provided below further illustrate and illustrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparations is not sufficient to limit the scope of the present invention in any way.

[0130] The following synthetic routes summarize and describe the preparation of the formula I derivatives of the present invention, all starting materials are prepared by the methods described in these routes, by methods well known to those skilled in the art of organic chemistry, or are commercially available. All final derivatives of the present invention are prepared by the methods described in these routes or by methods analogous thereto, which are well known to those skilled in the art of organic chemistry. All variables used in these routes are as defined below or as defined in the claims.

[0131] The derivatives of formula I according to the present invention can be prepared according to the methods of route A, route B, route C, route D, route E, route F, route G, route H, route I, route J and route K. Wherein, R1, R2, R3, R4, R5 and m in the compound are as defined in the claims.

[0132]

[0133] In route A, A1 is used as the starting material, and after esterification, ester aminolysis, amide dehydration, and reduction, it reacts with intermediate M6 to obtain intermediate A6, followed by cyanothioate addition, cyclization with 1,3-dichloroacetone, and nucleophilic substitution reaction with various substituted small molecular amines to obtain compound Ii of general formula I.

[0134]

[0135] In route B, the preparation of intermediate A6 is the same as route A. After esterification, ester aminolysis, amide dehydration, and reduction, it reacts with intermediate M6 to obtain intermediate A6, and then the cyano group is added with hydroxylamine hydrochloride, cyclized with chloroacetyl chloride, and then reacted with various substituted small molecular amines for nucleophilic substitution reaction to obtain compound I-ii of general formula I.

[0136] In route C, C1 is used as the starting material, which undergoes a click reaction with propargyl alcohol after diazotization and azidation, and then undergoes a nucleophilic substitution reaction with various substituted small molecule amines after chlorination to obtain intermediate C5, which is then reduced to the nitro group and undergoes a Buchwald–Hartwig coupling reaction with intermediate M6 to obtain compound I-iii of general formula I.

[0137]

[0138] In route D, A1 is used as the starting material, and after nitro reduction, it undergoes a nucleophilic substitution reaction with the intermediate M6, and then undergoes esterification, hydrazinolysis, and chloroacetic acid cyclization, and then undergoes a nucleophilic substitution reaction with various substituted small molecular amines to obtain compound I-iv of general formula I.

[0139]

[0140] In route E, A1 is used as the starting material, and after esterification, hydrazinolysis, chloroacetyl chloride substitution, and Lawesson reagent cyclization, it undergoes a nucleophilic substitution reaction with various substituted small molecule amines, and then the nitro group is reduced to undergo a Buchwald–Hartwig coupling reaction with the intermediate M6 to obtain compound Iv of general formula I.

[0141]

[0142] In route F, A1 is used as the starting material, and after nitro reduction, it undergoes a nucleophilic substitution reaction with the intermediate M6, and then undergoes esterification, aminolysis, and 1,3-dichloroacetone cyclization, and then undergoes a nucleophilic substitution reaction with various substituted small molecular amines to obtain compound I-Vi of general formula I.

[0143]

[0144] In route G, G1 is used as the starting material, and undergoes pyrrole substitution, nitro reduction, and nucleophilic substitution reaction with M6 to obtain G4, which is then subjected to Mannich reaction with various substituted small molecule amines to obtain compounds I-Vii of general formula I.

[0145]

[0146] In route H, H1 is used as the starting material, and after hydroxylamine addition, chlorination, propargyl alcohol cyclization, chlorination, and nucleophilic substitution reaction with various substituted small molecular amines, intermediate H6 is obtained, nitro reduction, and then nucleophilic substitution with M6 is obtained to obtain compounds I-Viii of general formula I.

[0147]

[0148] In route I, the preparation of intermediate B1 is the same as route B, and it is substituted with ethyl oxalyl chloride and then cyclized, and then hydrolyzed and condensed with various substituted small molecular amines to obtain compound I-iX of general formula I.

[0149]

[0150] In route J, the preparation of intermediate D1 is the same as route D, and it is substituted with ethyl oxalyl chloride and then cyclized, and then hydrolyzed and condensed with various substituted small molecular amines to obtain compound IX of general formula I.

[0151]

[0152] In route K, the preparation of intermediate E1 is the same as route E, and it is substituted with monoethyl oxalyl chloride and then cyclized, hydrolyzed and condensed with various substituted small molecular amines, and then the nitro group is reduced to undergo Buchwald-Hartwig coupling reaction with intermediate M6 to obtain compound I-Xi of general formula I.

[0153]

[0154] In route K, the preparation of intermediate C2 is the same as route C, and it undergoes a click reaction with propiolic acid, and then condenses with various substituted small molecule amines to obtain intermediate L2, which is then reduced to the nitro group and undergoes a Buchwald–Hartwig coupling reaction with intermediate M6 to obtain compound I-Xii of general formula I.

[0155]

[0156] In route M, H3 is used as the starting material, undergoes propiolic acid cyclization, and then undergoes condensation with various substituted small molecular amines to obtain intermediate M2, which is then nitro-reduced and then undergoes nucleophilic substitution with M6 to obtain compounds I-Xiii of general formula I.

[0157]

[0158] Intermediate M6 can be obtained by using M1 as the starting material according to route M, reacting with 2,4-dichloro-5-nitropyrimidine after esterification and reductive amination, then reducing and cyclizing, and finally undergoing nucleophilic substitution reaction. Wherein, R1 and R2 in the compound are as defined in the claim. DETAILED DESCRIPTION

[0159] In the following examples, methods for preparing some of the compounds are described. It should be understood that the following methods and other methods known to those of ordinary skill in the art can be applied to the preparation of all compounds described in the present invention. The examples are intended to illustrate rather than limit the scope of the present invention. The H NMR spectra of the compounds were measured using Bruker ARX-400 or Bruker ARX-600, and the mass spectra were measured using Agilent 1100 LC / MSD; all reagents used were analytically pure or chemically pure.

[0160] Example 1: (R)-8-cyclopentyl-2-((4-(4-((dimethylamino)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0161] Step A Synthesis of methyl 3-methoxy-4-nitrobenzoate (A2)

[0162] 197.1g (1.00mol) of 3-methoxy-4-nitrobenzoic acid was added to 2000mL of dry methanol, and 201.9g (1.80mol) of thionyl chloride was slowly added dropwise at 0°C. After the addition was completed, the temperature was raised to room temperature and the reaction was continued for 10h. The reaction solution was concentrated and poured into a large amount of water. The pH value was adjusted to neutral with a saturated sodium bicarbonate aqueous solution. After suction filtration and drying, 187.9g of a light yellow solid was obtained, with a yield of 89.0%. MS (ESI) m / z: 212.1 [M+H] + .

[0163] Step B Synthesis of 3-methoxy-4-nitrobenzamide (A3)

[0164] 105.6g (0.50mol) of methyl 3-methoxy-4-nitrobenzoate was added to 500mL of methanol, and 1L of concentrated ammonia was slowly added. The temperature was raised to 50°C and the reaction was continued for 4h. After cooling to room temperature, the reaction solution was concentrated and poured into a large amount of water. The mixture was stirred at room temperature for 30min, filtered and dried to obtain 75.5g of a light yellow solid with a yield of 77%. MS (ESI) m / z: 197.1 [M+H] + .

[0165] Step C Synthesis of 3-methoxy-4-nitrobenzonitrile (A4)

[0166] 68.6 g (0.35 mol) of 3-methoxy-4-nitrobenzamide was added to 300 mL of tetrahydrofuran, and 70.8 g (0.70 mol) of triethylamine was added. 88.2 g (0.42 mol) of trifluoroacetic anhydride was slowly added dropwise at 0°C, and the temperature was raised to room temperature, and the reaction was carried out for 3 hours. Most of the solvent was evaporated, and the residue was poured into water, stirred at room temperature for 30 minutes, filtered, and the filter cake was dried to obtain 51.1 g of yellow solid, with a yield of 82%, MS (ESI) m / z: 179.1 [M+H] + .

[0167] Step D Synthesis of 3-methoxy-4-aminobenzonitrile (A5)

[0168] Add 61.6g (1.10mol) of reduced iron powder to 400mL of anhydrous ethanol in batches, slowly drop 3mL of 37% hydrochloric acid, stir at 50°C for 30min, then add 35.6g (0.20mol) of 3-methoxy-4-nitrobenzonitrile, and react at 80°C for 4h. Filter with diatomaceous earth pad while hot, evaporate the filtrate to dryness, and obtain 16.05g of brown-red solid, with a yield of 54.2%, MS (ESI) m / z: 149.1 [M+H] + .

[0169] Step E Synthesis of (R)-2-aminobutyric acid methyl ester hydrochloride (M2)

[0170] 200g (1.94mol) (R)-2-aminobutyric acid was added to 1000mL methanol, 391.8g (3.49mol) thionyl chloride was slowly added dropwise at -10°C, the temperature was raised to 70°C and refluxed, and the reaction was carried out for 10h. The reaction solution was evaporated to dryness to obtain a light yellow oily substance. After cooling, methyl tert-butyl ether was added and stirred. A white solid was generated and filtered. 289.0g of white solid was obtained, with a yield of 97%. MS (ESI) m / z: 118.2 [M+H] + .

[0171] Step F Synthesis of (R)-2-(cyclopentylamino)butyric acid methyl ester (M3)

[0172] 229.6 g (1.50 mol) of (R)-2-aminobutyric acid methyl ester hydrochloride was dissolved in 2500 mL of DCM, and 138.7 g (1.65 mol) of cyclopentanone and 123.0 g (1.50 mol) of anhydrous sodium acetate were added. After reacting for 1.5 h, 349.7 g (1.65 mol) of sodium triacetoxyborohydride were added in batches. After reacting at room temperature for 4 h, the solution was adjusted to weak alkalinity, extracted with DCM, dried, and the solvent was evaporated under reduced pressure to obtain 249.9 g of a light yellow oil with a yield of 90%. MS (ESI) m / z: 186.1 [M+H] + ; 1H NMR (600MHz, Chloroform-d) δ3.65 (s, 1H), 1.75–1.64 (m, 1H), 1.65–1.53 (m, 1H), 0.84 (t, J = 7.5Hz, 1H).

[0173] Step G Synthesis of (R)-2-[(2-chloro-5-nitro-4-pyrimidinyl)-cyclopentylamino]butyric acid methyl ester (M4)

[0174] 231.4g (1.25mol) (R)-2-(cyclopentylamino)butyric acid methyl ester and 105.0g (1.25mol) sodium bicarbonate were added to 2000mL cyclohexane, and 193.9g (1.00mol) 2,4-dichloro-5-nitropyrimidine was added at 0°C, and the temperature was raised to 80°C for reaction for 10h. After the reaction solution was cooled to room temperature, the solid residue was removed by suction filtration, and the solvent was removed by vacuum distillation to obtain a reddish brown crude product. After column chromatography purification, a light yellow solid was obtained, 239.9g, yield 70%, mp: 109.7–110.5°C, MS (ESI) m / z: 343.1 [M+H] + ; 1 HNMR(600MHz,Chloroform-d)δ8.59(s,1H),3.68(s,2H),3.52–3.44(m,1H),2.34(dt,J=14.1,7.2 Hz,1H),2.14(dt,J=7.7,4.1Hz,1H),1.99–1.83(m,1H),1.75–1.40(m,4H),0.98(t,J=7.5Hz,2H).

[0175] Step H Synthesis of (R)-2-chloro-8-cyclopentyl-7-ethyl-7,8-dihydro-6(5H)-pteridinone (M5)

[0176] 171.4 g (0.50 mol) of (2R)-2-[(2-chloro-5-nitro-4-pyrimidinyl)cyclopentylamino]-butyric acid methyl ester was dissolved in 2000 mL of glacial acetic acid, and 156.8 g (2.80 mol) of iron powder was added in batches at 70 °C. After reacting for 0.5 h, the mixture was filtered with diatomaceous earth pad while hot, and the filtrate was evaporated to dryness to obtain 130.8 g of white solid with a yield of 93.2%. mp: 171.8–172.7 °C, MS (ESI) m / z: 281.1 [M+H] + ; 1H NMR (600MHz, Chloroform-d) δ9.49(s,1H),7.61(s,1H),4.25(s,1H),2.03(s,1H),1.81(s,3H),1.74–1.67(m,1H),1.62–1.53(m,1H),0.87(s,2H).

[0177] Step I Synthesis of (R)-2-chloro-8-cyclopentyl-7-ethyl-7,8-dihydro-5-methyl-6(5H)-pteridinone (M6)

[0178] 126.3 g (0.45 mol) (R)-2-chloro-8-cyclopentyl-7-ethyl-7,8-dihydro-6 (5H) -pteridinone was dissolved in 1000 mL N, N-dimethylformamide (DMF), and 10.8 g (0.45 mol) sodium hydride was added at -10 ° C. After stirring for 1 hour, 70.3 g (0.495 mol) iodomethane was slowly added and reacted at room temperature for 10 hours. The reaction solution was then poured into water, filtered and dried to obtain 118.9 g of a white solid with a yield of 89.7%, mp: 92.1-93.5 ° C, MS (ESI) m / z: 295.1 [M+H] + ; 1 H NMR(600MHz,Chloroform-d)δ4.27(p,J=8.1Hz,1H),4.20(dd,J=7.5,3.5Hz,1H),3.26(s,2H),2. 05–1.98(m,1H),1.94–1.76(m,3H),1.75–1.63(m,1H),1.63–1.55(m,1H),0.79(t,J=7.5Hz,2H).

[0179] Step J Synthesis of (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropyridin-2-yl)amino)-3-methoxybenzonitrile (A6)

[0180] 13.4 g (0.10 mol) of 3-methoxy-4-aminobenzonitrile was added to 500 mL of isopropanol, and 38.3 g (0.13 mol) of (R)-2-chloro-8-cyclopentyl-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (M6) was added. 10 mL of 37% hydrochloric acid was slowly added, and the mixture was reacted at 90°C for 10 h. Most of the solvent was evaporated, and the residue was added with 100 mL of ethyl acetate and stirred for 30 min, and then filtered to obtain 37.8 g of a white solid with a yield of 93.1%. MS (ESI) m / z: 407.2 [M+H] + .

[0181] Step K Synthesis of (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-3-methoxybenzothioamide (A7)

[0182] 20g (0.25mol) of 70% sodium hydrosulfide and 50g (0.25mol) of magnesium chloride hexahydrate were added to 1000mL of N,N-dimethylformamide (DMF), and 20.3g (0.05mol) of intermediate A6 were added in batches. The mixture was reacted at room temperature for 4h. After removing the insoluble matter by suction filtration, the filtrate was poured into water to generate solids. After suction filtration again, 19.2g of yellow solids were obtained, with a yield of 87.1%. MS (ESI) m / z: 441.2 [M+H] + .

[0183] Step L Synthesis of (R)-2-((4-(4-(chloromethyl)thiazol-2-yl)-2-methoxyphenyl)amino)-8-cyclopentyl-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (A8)

[0184] 4.48 g (0.08 mol) of 1,3-dichloroacetone and 16.2 g (0.04 mol) of (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-3-methoxybenzothioamide were added to 200 ml of toluene, and the temperature was raised to 90°C for reaction for 10 h. The solvent was evaporated and purified by column chromatography to obtain 15.4 g of a reddish brown solid with a yield of 75.7%. MS (ESI) m / z: 513.2 [M+H] + .

[0185] Step M Synthesis of (R)-8-cyclopentyl-2-((4-(4-((dimethylamino)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0186] 1 g (1.97 mmol) of intermediate A8, 0.55 g (3.94 mmol) of potassium carbonate and 0.27 g of dimethylamine aqueous solution (2.36 mmol) were dissolved in 15 mL of dry N,N-dimethylformamide (DMF), stirred at room temperature for 4 h, and the reaction solution was poured into water to produce a large amount of solid, which was filtered and dried, and then purified by column chromatography to obtain 0.8 g of light yellow solid with a yield of 70.8%. mp: 198.2–200.5°C; MS (ESI) m / z: 522.68 [M+H ]+ ; 1H NMR(600MHz,DMSO-d6)δ8.45(d,J=8.4Hz,1H),7.84(s,1H),7.63(s,1H),7.51(s,1H) ,7.49(d,J=1.8Hz,1H),7.48(dd,J=8.4,1.8Hz,1H),4.35(p,J=8.4Hz,1H),4.22(dd, J=7.7,3.6Hz,1H),3.97(s,3H),3.74(s,2H),3.24(s,3H),2.34(s,6H),2.04–1.98(m ,1H),1.89–1.82(m,2H),1.80–1.70(m,4H),1.67–1.55(m,3H),0.76(t,J=7.5Hz,3H).

[0187] According to the method of Example 1, the intermediate A8 of Example 1 was used as a raw material to carry out nucleophilic substitution reaction with the corresponding small molecule amine to prepare the compounds of Examples 2 to 7. The structures, melting points, MS and 1H-NMR data of some of the compounds of the examples are shown in Table 1.

[0188] Table 1 Example 1 to Example 7

[0189]

[0190]

[0191] Example 8: (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(piperazin-1-ylmethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0192] Step A Synthesis of (R, Z)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-N'-hydroxy-3-methoxybenzamide (B1)

[0193] 20.3 g (0.05 mol) of intermediate A6 of Example 1, 5.21 g (0.075 mol) of hydroxylamine hydrochloride and 13.8 g (0.10 mol) of potassium carbonate were added to methanol, and the temperature was raised to 70°C under nitrogen protection for 6.5 h. After the insoluble matter was removed by suction filtration, the filtrate was evaporated to dryness to obtain 21.1 g of white solid, with a yield of 96.3%. MS (ESI) m / z: 440.2 [M+H] + .

[0194] Step B Synthesis of (R)-2-((4-(5-(chloromethyl)-1,2,4-oxadiazol-3-yl)-2-methoxyphenyl)amino)-8-cyclopentyl-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (B2)

[0195] 19.7 g (0.045 mol) (R, Z)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-N'-hydroxy-3-methoxybenzamide and 7.62 g (0.0675 mol) chloroacetyl chloride were added to 200 mL toluene, and the temperature was raised to 90°C for reaction for 10 h. After evaporating the solution, column chromatography was performed to purify the solution to obtain 18.5 g of a light yellow solid with a yield of 82.7%. MS (ESI) m / z: 498.9 [M+H] + .

[0196] Step C Synthesis of (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(piperazin-1-ylmethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0197] 1g (2.00mmol) of intermediate B3, 0.55g (4.00mmol) of potassium carbonate and 0.24g of piperazine (2.40mmol) were dissolved in 15mL of dry N,N-dimethylformamide (DMF), stirred at room temperature for 4h, and the reaction solution was poured into water to produce a large amount of solid, which was filtered and dried, and then purified by column chromatography to obtain 0.92g of light yellow solid, with a yield of 82.3%. MS (ESI) m / z: 548.7 [M+H] +1 ; 1 H NMR (600MHz, DMSO-d6) δ8.57(d,J=8.4Hz,1H),7.87(s,1H),7.70(s,1H),7.63(dd,J=8. 4,1.7Hz,1H),7.55(d,J=1.8Hz,1H),4.41–4.34(m,1H),4.25(dd,J=7.6,3.6Hz,1H),4. 04(s,2H),3.99(s,3H),3.26(s,4H),3.03–2.98(m,4H),2.74–2.68(m,3H),2.06–2.00( m,1H),1.95–1.85(m,2H),1.82–1.74(m,4H),1.69–1.60(m,3H),0.77(t,J=7.5Hz,3H).

[0198] According to the method of Example 8, the intermediate B2 of Example 8 was used as a raw material to carry out nucleophilic substitution reaction with the corresponding small molecular amine to prepare the compounds of Examples 9 to 16. The structures, chemical names and melting points, MS, 1 The H-NMR data are shown in Table 2.

[0199] Table 2 Example 8 to Example 16

[0200]

[0201]

[0202]

[0203] Example 17: (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(4-((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0204] Step A Synthesis of 4-azido-2-methoxy-1-nitrobenzene (C2)

[0205] Suspend 16.8g (0.1mol) 3-methoxy-4-nitroaniline in 200mL 37% hydrochloric acid, cool to below 0℃, and stir at low temperature for 30min. Dissolve 7.58g (0.11mol) sodium nitrite in 20mL water, slowly drop it into the above reaction solution, keep the temperature below 5℃, and continue to react at low temperature for 30min. Dissolve 6.5g (0.1mol) sodium azide in 10mL water, slowly drop it into the above reaction solution, and keep the temperature below 5℃ during the addition process. After the addition is complete, react at 0℃ for 30min, then warm to room temperature and react for 3h. Filter and dry to obtain 15.14g of light yellow solid, with a yield of 78.1%.

[0206] Step B Synthesis of (1-(3-methoxy-4-nitrophenyl)-1H-1,2,3-triazol-4-yl)methanol (C3)

[0207] At 0°C, 9.7g (0.05mol) 4-azido-2-methoxy-1-nitrobenzene, 4.57g (0.024mol) cuprous iodide, 2.38g (0.012mol) sodium vitamin C and 3.08g (0.055mol) propargyl alcohol were added to 100mL DMF in sequence, and the reaction was continued for 30min under ice bath, and then the temperature was raised to room temperature and the reaction was continued for 3h. After the reaction was completed, the mixture was poured into water and then extracted with dichloromethane. The organic phase was washed twice with saturated brine, and the solvent was evaporated to obtain 9.65g of light yellow oil, with a yield of 77.1%, MS (ESI) m / z: 251.1 [M+H] +1 .

[0208] Step C Synthesis of 4-(chloromethyl)-1-(3-methoxy-4-nitrophenyl)-1H-1,2,3-triazole (C4)

[0209] 8.75g (0.035mol) (1-(3-methoxy-4-nitrophenyl)-1H-1,2,3-triazol-4-yl) methanol was added to 100mL dichloromethane, and 8.33g (0.07mol) thionyl chloride was slowly added dropwise, and the temperature was raised to reflux, and the reaction was carried out for 2h. After the reaction was completed, the reaction solution was concentrated and poured into water, filtered, and dried to obtain 7.52g of light yellow solid, with a yield of 80%, MS (ESI) m / z: 269.1 [M+H] +1 .

[0210] Step D Synthesis of 1-((1-(3-methoxy-4-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)-4-methylpiperazine (C5)

[0211] 6.72 g (0.025 mol) of 4-(chloromethyl)-1-(3-methoxy-4-nitrophenyl)-1H-1,2,3-triazole, 6.9 g (0.05 mol) of potassium carbonate and 3 g of N-methylpiperazine (0.03 mol) were dissolved in 50 mL of dry N,N-dimethylformamide (DMF) and stirred at room temperature for 4 h. The reaction solution was poured into water to produce a large amount of solids. The solids were filtered and dried to obtain 7.36 g of light yellow solids with a yield of 88.6%. MS (ESI) m / z: 333.1 [M+H] +1 .

[0212] Step E Synthesis of 2-methoxy-4-(4-(((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)aniline (C6)

[0213] 6g (0.018mol) of 1-((1-(3-methoxy-4-nitrophenyl)-1H-1,2,3-triazol-4-yl)methyl)-4-methylpiperazine and 0.6g of palladium on carbon (Pd / C) were added to 50mL of methanol, hydrogen was introduced, vacuum was drawn, and the reaction was carried out at room temperature for 6h. Filtered by suction, the filter cake was washed with methanol (30mL×3), the filter cake was discarded, and the filtrate was retained. The solvent of the filtrate was removed to obtain 5.08g of black oil, with a yield of 92.6%, MS (ESI) m / z: 303.2 [M+H] +1 .

[0214] Step F Synthesis of (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(4-((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0215] Under nitrogen protection, (0.19 g, 0.65 mmol) 2-methoxy-4-(4-(((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)aniline, (0.23 g, 0.78 mmol) intermediate M6, (0.03 g, 0.03 mmol) Pd2(dba)3, (0.046 g, 0.15 mmol) X-Phos, (0.84 g, 2.6 mmol) cesium carbonate were added to 10 mL of tert-butanol and reacted at 90 ° C for 12 hours. The reaction solution was filtered with diatomaceous earth, the filtrate was evaporated to dryness, and purified by column chromatography to obtain 0.1 g of light yellow solid with a yield of 28%. MS (ESI) m / z: 561.7 [M+H] + ; 1 H NMR(600MHz,DMSO-d6)δ8.74(s,1H),8.51(d,J=8.7Hz,1H),7.90(s,1H),7.66(s,1H),7. 59(s,1H),7.52(d,J=8.5Hz,1H),4.39(dd,J=16.6,8.3Hz,1H),4.33–4.26(m,1H),4.04(s ,3H),3.68(s,2H),3.31(s,3H),2.45(s,8H),2.23(s,3H),2.06(d,J=7.7Hz,1H),1.94(d d,J=15.8,10.3Hz,2H),1.83(t,J=15.4Hz,4H),1.74–1.59(m,3H),0.82(t,J=7.3Hz,3H).

[0216] According to the method of Example 17, the intermediate C4 of Example 17 was used as a raw material to carry out a nucleophilic substitution reaction with the corresponding small molecule amine, and then the nitro group was reduced and then reacted with the intermediate M6 to prepare the compounds of Examples 18 to 25. The structures, chemical names and melting points, MS, 1 The H-NMR data are shown in Table 3.

[0217] Table 3 Example 18 to Example 25

[0218]

[0219]

[0220]

[0221] Example 26: (R)-8-Cyclopentyl-2-((4-(5-((dimethylamino)methyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0222] Step A Synthesis of 4-amino-3-methoxybenzoic acid (D1)

[0223] Under hydrogen atmosphere, 10g of 3-methoxy-4-nitrobenzoic acid and 0.5g of palladium carbon were added to 100mL of methanol and reacted at room temperature for 12 hours. The reaction solution was filtered through diatomaceous earth to remove palladium carbon, and the filtrate was evaporated to dryness to obtain 8.5g of gray solid, with a yield of 92%, MS (ESI) m / z: 168.1 [M+H] +1 .

[0224] Step B Synthesis of (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropyridin-2-yl)amino)-3-methoxybenzoic acid (D2)

[0225] 2.75 g of 4-amino-3-methoxybenzoic acid was added to 25 mL of isopropanol, and 4.62 g of (R)-2-chloro-8-cyclopentyl-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (M6) was added. 2.7 mL of 37% hydrochloric acid was slowly added, and the mixture was reacted at 90°C for 10 h. Most of the solvent was evaporated, and the residue was added with 100 mL of ethyl acetate and stirred for 30 min, and then filtered to obtain 3.6 g of a white solid with a yield of 54%. MS (ESI) m / z: 426.2 [M+H] +1 .

[0226] Step C Synthesis of (R)-methyl 4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)methyl)-3-methoxybenzoate (D3)

[0227] 3.6 g (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropyridin-2-yl)amino)-3-methoxybenzoic acid (D2) and 1.79 g sodium carbonate were added to 18 mL DMF, and 1.44 g iodomethane was added dropwise. The reaction was carried out at room temperature for 5-6 hours. The reaction solution was poured into water and filtered to obtain 3.16 g of a white solid with a yield of 85%. MS (ESI) m / z: 440.2 [M+H] +1 .

[0228] Step D Synthesis of (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-3-methoxybenzohydrazide (D4)

[0229] 3.0 g of (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)methyl)-3-methoxybenzoic acid methyl ester (D3) was added to a mixed solvent of 20 mL of hydrazine hydrate and 10 mL of ethanol, and the mixture was reacted at 100°C for 12 h. After the reaction solution was cooled, a large amount of solid precipitated, which was filtered and dried to obtain 2.87 g of a white solid with a yield of 90%. MS (ESI) m / z: 440.2 [M+H] +1 .

[0230] Step E Synthesis of (R)-2-((4-(5-(chloromethyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-8-cyclopentyl-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (D5)

[0231] 2.87g (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-3-methoxybenzoic acid hydrazide (D4) and 1.23g chloroacetic acid were added to 20ml phosphorus oxychloride, and the temperature was raised to 105°C for reaction for 10-12h. The reaction solution was concentrated and added to ice water, and the pH was adjusted to neutral. It was then extracted with ethyl acetate, the organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. After column chromatography, 2.3g of yellow solid was obtained, with a yield of 70%, MS (ESI) m / z: 498.2 [M+H] +1 .

[0232] Step F Synthesis of (R)-8-cyclopentyl-2-((4-(5-((dimethylamino)methyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one

[0233] 1 g (2.00 mmol) of intermediate D5, 0.55 g (4.00 mmol) of potassium carbonate and 0.25 g of dimethylamine hydrochloride (2.40 mmol) were dissolved in 15 mL of dry N,N-dimethylformamide (DMF), and stirred at room temperature for 4 h. The reaction solution was poured into water to produce a large amount of solid, which was filtered and dried. After purification by column chromatography, 0.86 g of light yellow solid was obtained. The yield was 77%. MS (ESI) m / z: 507.61 [M+H] + ;

[0234] According to the method of Example 26, the intermediate D5 of Example 26 was used as a raw material to carry out nucleophilic substitution reaction with the corresponding small molecule amine to prepare the compounds of Examples 27 to 38. The structures, chemical names and melting points, MS, and 1H-NMR data of some of the compounds of the examples are shown in Table 4.

[0235] Table 4 Examples 26 to 38

[0236]

[0237]

[0238]

[0239]

[0240] According to the synthesis method of intermediate M6 in Example 1, the corresponding dihydropteridinone skeleton was synthesized by using the corresponding aldehyde or ketone instead of cyclopentanone as the raw material, and then the compounds of Examples 39 to 50 were synthesized with the corresponding small molecule amine according to the method of Example 26. The structures, chemical names and melting points, MS, 1 The H-NMR data are shown in Table 5.

[0241] Table 5 Example 39 to Example 50

[0242]

[0243]

[0244]

[0245]

[0246] Example 51: (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((3-methylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0247] Step A: Synthesis of 3-methoxy-4-nitrobenzoylhydrazide (G1)

[0248] 21.1g (0.10mol) of methyl 3-methoxy-4-nitrobenzoate was dissolved in 80mL of methanol, and then 100mL of 80% hydrazine hydrate was added and the temperature was raised to 50°C for 1h. The reaction solution was concentrated and added to water, filtered, and the filter cake was dried to obtain 14.7g of white solid, with a yield of 70%. MS (ESI) m / z: 212.2 [M+H] + .

[0249] Step B: Synthesis of N'-(2-chloroacetyl)-3-methoxy-4-nitrobenzoylhydrazide (G2)

[0250] 14g 3-methoxy-4-nitrobenzoyl hydrazide and 8.9g chloroacetyl chloride were added to 100ml acetonitrile, cooled to 0℃, and 3.2mL 50% sodium hydroxide aqueous solution was added to react for 2h. The reaction solution was concentrated and added to water, filtered, and the filter cake was dried to obtain 10.2g of white solid, with a yield of 50%. MS (ESI) m / z: 287.9 ​​[M+H] + .

[0251] Step C: Synthesis of 2-(chloromethyl)-5-(3-methoxy-4-nitrophenyl)-1,3,4-thiadiazole (G3)

[0252] Dissolve 10 g of N'-(2-chloroacetyl)-3-methoxy-4-nitrobenzoyl hydrazide in 100 mL of THF, add 8.4 g of Lawesson's reagent, and then heat to 80°C for 10 h. After TLC detection, pour the reaction solution into a saturated sodium carbonate aqueous solution, extract with ethyl acetate, wash the organic phase with a saturated sodium chloride solution, and purify by column chromatography to obtain 5.2 g of a yellow solid, with a yield of 52%. MS (ESI) m / z: 285.9 [M+H] + .

[0253] Step D: Synthesis of 2-(3-methoxy-4-nitrophenyl)-5-((3-methylpiperazin-1-yl)methyl)-1,3,4-thiadiazole (G4)

[0254] 0.5 g (1.75 mmol) 2-(chloromethyl)-5-(3-methoxy-4-nitrophenyl)-1,3,4-thiadiazole, 0.48 g (2.00 mmol) potassium carbonate and 0.875 g 3-methylpiperazine (8.75 mmol) were dissolved in 5 mL N,N-dimethylformamide (DMF) and stirred at room temperature for 4 h. The reaction solution was poured into water and extracted with ethyl acetate. After purification by column chromatography, 0.57 g of a yellow solid was obtained. The yield was 93%. MS (ESI) m / z: 350.1 [M+H] + .

[0255] Step E: Synthesis of 2-methoxy-4-(5-((3-methylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl)aniline (G4)

[0256] 0.5g 2-(3-methoxy-4-nitrophenyl)-5-((3-methylpiperazine-1-yl)methyl)-1,3,4-thiadiazole and 0.5g reduced iron powder were added to 20mL ethanol, and then heated to 80℃ for 4h. The reaction solution was filtered while hot with diatomaceous earth and evaporated to dryness to obtain 0.29g of black solid, with a yield of 61%, MS (ESI) m / z: 320.1 [M+H] + .

[0257] Step F: Synthesis of (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((3-methylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0258] Under nitrogen protection, (0.21g, 0.65mmol) 2-methoxy-4-(5-((3-methylpiperazine-1-yl)methyl)-1,3,4-thiadiazol-2-yl)aniline, (0.23g, 0.78mmol) intermediate M6, (0.03g, 0.03mmol) Pd2(dba)3, (0.046g, 0.15mmol) X-Phos, (0.84g, 2.6mmol) cesium carbonate were added to 10mL of tert-butanol and reacted at 90°C for 12 hours. The reaction solution was filtered with diatomaceous earth pad, the filtrate was evaporated to dryness, and purified by column chromatography to obtain 0.14g of light yellow solid, with a yield of 37%, MS (ESI) m / z: 578.3 [M+H] + ; 1H NMR (600MHz, CDCl3) δ8.54(d,J=8.4Hz,1H),7.62(s,1H),7.55(d,J=8.4Hz,2H),7.35(d,J=8.3Hz,1H),4.49– 4.41(m,1H),4.16(dd,J=7.8,3.6Hz,1H),3.94(s,3H),3.90(d,J=4.2Hz,2H),3.27(s,3H),3.04(d,J=12.0Hz ,1H),2.99–2.89(m,2H),2.83(d,J=11.0Hz,2H),2.33(t,J=11.2Hz,1H),2.11–2.06(m,1H),2.01(t,J=10.5H z,1H),1.97–1.92(m,1H),1.83–1.74(m,4H),1.70–1.61(m,4H),1.08(d,J=6.3Hz,3H),0.81(t,J=7.5Hz,3H).

[0259] According to the synthesis method of Example 51, the compounds of Examples 52 to 55 were synthesized using the corresponding small molecule amines. The structures, chemical names, melting points, MS, 1 H-NMR data are shown in Table 6

[0260] Table 6 Example 51 to Example 55

[0261]

[0262]

[0263] Example 56 (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(piperazin-1-ylmethyl)oxazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0264] Step A Synthesis of (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)methyl)-3-methoxybenzamide (F1)

[0265] To a mixed solvent of 15 mL of aqueous ammonia and 3 mL of methanol was added 3 g of (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)methyl)-3-methoxybenzoic acid methyl ester (D3) and reacted at 50°C for 3 h. After the reaction solution was cooled, a large amount of solid precipitated, which was filtered and dried to obtain 2.67 g of a light yellow solid with a yield of 87%. MS (ESI) m / z: 425.2 [M+H] + .

[0266] Step B Synthesis of (R)-2-((4-(5-(chloromethyl)oxazol-2-yl)-2-methoxyphenyl)amino)-8-cyclopentyl-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (F2)

[0267] 2.67 g (R)-4-(((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-3-methoxybenzamide (F1) and 1.60 g 1,3-dichloroacetone were added to 20 ml toluene, and the temperature was raised to 105°C for reaction for 10-12 h. After the reaction, the reaction solution was evaporated and column chromatography was performed to obtain 2.42 g of a light yellow solid with a yield of 70%. MS (ESI) m / z: 497.2 [M+H] + .

[0268] Step C Synthesis of (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(piperazin-1-ylmethyl)oxazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0269] 0.2 g (0.4 mmol) of intermediate F2, 0.01 g (0.08 mmol) of potassium carbonate and 0.02 g of anhydrous piperazine (0.2 mmol) were dissolved in 5 mL of dry N,N-dimethylformamide (DMF) and stirred at room temperature for 4 h. The reaction solution was poured into water, extracted with ethyl acetate, and purified by column chromatography to obtain 0.13 g of a yellow solid with a yield of 65%. MS (ESI) m / z: 547.3 [M+H] + .

[0270] According to the synthesis method of Example 56, the compounds of Examples 57 to 59 were synthesized using the corresponding small molecule amines. The structures, chemical names, melting points, MS, 1 H-NMR data are shown in Table 7.

[0271] Table 7 Example 56 to Example 59

[0272]

[0273]

[0274] Example 60: (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(2-(((4-methylpiperazin-1-yl)methyl)-1H-pyrrol-1-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0275] Step A Synthesis of 1-(3-methoxy-4-nitrophenyl)-1H-pyrrole (G2)

[0276] At room temperature, 4-fluoro-2-methoxy-1-nitrobenzene (20.0 g, 0.12 mol) was dissolved in N,N-dimethylformamide (DMF) (200 mL), potassium carbonate (33.2 g, 0.24 mol) and pyrrole (8.9 g, 0.13 mol) were added under stirring, and the temperature was raised to 100°C for reaction for 12 h. After the reaction, the reaction solution was poured into water (1000 mL), extracted with ethyl acetate three times, the organic layers were combined, washed with water and saturated brine three times in sequence, dried over anhydrous Na2SO4, filtered, and the solvent was evaporated under reduced pressure to obtain 22.9 g of a yellow solid with a yield of 87.6%. MS (ESI) m / z: 219.1 [M+H] + ;.

[0277] Step B Synthesis of 2-methoxy-4-(1H-pyrrol-1-yl)aniline (G3)

[0278] At room temperature, iron powder (20.2 g, 0.36 mol) was added to 95% ethanol (200 mL) under stirring, and the temperature was raised to 70°C for activation reaction for 30 min. 1-(3-methoxy-4-nitrophenyl)-1H-pyrrole (20.0 g, 0.09 mol) was added to the reaction solution, and the temperature was raised to 80°C for reaction for 5 h. After the reaction, diatomaceous earth was pad and filtered while hot, and most of the solvent in the organic layer was evaporated to obtain 17.3 g of black oil, with a yield of 85.4%, MS (ESI) m / z: 189.1 [M+H] + ;.

[0279] Step C Synthesis of (R)-8-cyclopentyl-7-ethyl-2-(((2-methoxy-4-(1H-pyrrol-1-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (G4)

[0280] 11.3g (0.06mol) of 2-methoxy-4-(1H-pyrrol-1-yl)aniline was added to 500mL of isopropanol, and 22.9g (0.078mol) of (R)-2-chloro-8-cyclopentyl-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (M6) was added. 10mL of 37% hydrochloric acid was slowly added, and the mixture was reacted at 90°C for 10h. Most of the solvent was evaporated, and the residue was added with 100mL of ethyl acetate and stirred for 30min, and then filtered to obtain 22.4g of white solid, with a yield of 83.6%, and MS (ESI) m / z: 447.3 [M+H] + ;.

[0281] Step D Synthesis of (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(2-(((4-methylpiperazin-1-yl)methyl)-1H-pyrrol-1-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0282] N-methylpiperazine (0.04 g) and formaldehyde aqueous solution (0.15 mL) were dissolved in glacial acetic acid (5 mL) and stirred for half an hour. Intermediate G4 (0.2 g) dissolved in glacial acetic acid (5 mL) was slowly dripped into the reaction solution and reacted for 5 hours. After the reaction, most of the solvent was evaporated under reduced pressure, water (5 mL) was added, stirred, saturated sodium bicarbonate aqueous solution was adjusted to pH 8-9, extracted twice with dichloromethane, the organic layers were combined, washed twice with water and saturated brine in turn, evaporated to dryness, and column chromatography was performed to obtain 60 mg of a white solid with a yield of 25.0%. ESI-MS [M+H] (m / z): 559.3

[0283] According to the synthesis method of Example 60, the compounds of Examples 61 to 64 were synthesized using the corresponding small molecule amines. The structures, chemical names, melting points, MS, 1 H-NMR data are shown in Table 8

[0284] Table 8 Example 60 to Example 64

[0285]

[0286]

[0287] Example 65: (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((4-methylpiperazin-1-yl)methyl)isoxazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropyridin-6(5H)-one

[0288] Step A: Synthesis of 3-methoxy-4-nitrobenzaldehyde oxime (H2)

[0289] Dissolve 50g of 3-methoxy-4-nitrobenzaldehyde in methanol, add 34.5g of hydroxylamine hydrochloride, and heat to 70°C for 4h. After the reaction is completed, evaporate the methanol under reduced pressure. Dissolve the residue in ethyl acetate, wash with water and brine, dry with sodium sulfate, and remove the solvent under reduced pressure. The obtained solid is recrystallized from ethyl acetate and petroleum ether to obtain 50g of off-white solid with a yield of 91%.

[0290] Step B: Synthesis of N-hydroxy-3-methoxy-4-nitrobenzoyl chloride (H3)

[0291] 52.3 g of N-chlorosuccinimide was added to a solution of 50 g of 3-methoxy-4-nitrobenzaldehyde oxime dissolved in DMF (250 ml), and stirred at room temperature for 5 h. After the reaction was completed, the reaction solution was poured into water, extracted with ethyl acetate, washed with water, dried with sodium sulfate, and the solvent was removed under reduced pressure to obtain a crude product. The product was purified by column chromatography using petroleum ether / ethyl acetate: 1:9 to obtain 51 g of a white solid with a yield of 84%.

[0292] Step C: Synthesis of (3-(3-methoxy-4-nitrophenyl)isoxazol-5-yl)methanol (H4)

[0293] Dissolve 10 g of N-hydroxy-3-methoxy-4-nitrobenzoyl chloride and 2.28 g of propargyl alcohol in 120 ml of a 1:1 mixed solvent of tert-butyl alcohol and water. Add 0.76 g of VcNa, 0.2 g of copper sulfate pentahydrate and 16 g of potassium bicarbonate to the reaction solution under stirring. Stir at room temperature for 1 h. Dilute with water and filter to obtain 7.6 g of an off-white solid with a yield of 70%.

[0294] Step D: Synthesis of 5-(chloromethyl)-3-(3-methoxy-4-nitrophenyl)isoxazole (H5)

[0295] 7.6 g (3-(3-methoxy-4-nitrophenyl)isoxazol-5-yl)methanol was added to 100 mL dichloromethane, 6.5 g thionyl chloride was slowly added dropwise, the temperature was raised to reflux, and the reaction was carried out for 2 h. After the reaction was completed, the reaction solution was concentrated and poured into water, filtered, and dried to obtain 6.7 g of light yellow solid, with a yield of 76%, MS (ESI) m / z: 269.1 [M+H] +1 .

[0296] Step E: Synthesis of 3-(3-methoxy-4-nitrophenyl)-5-((4-methylpiperazin-1-yl)methyl)isoxazole (H6)

[0297] 6.7 g 5-(chloromethyl)-3-(3-methoxy-4-nitrophenyl)isoxazole, 6.9 g potassium carbonate and 3 g N-methylpiperazine were dissolved in 50 mL dry N,N-dimethylformamide (DMF) and stirred at room temperature for 4 h. The reaction solution was poured into water to produce a large amount of solids. The solids were filtered and dried to obtain 7.36 g of light yellow solids with a yield of 88.6%. MS (ESI) m / z: 333.1 [M+H] +1 .

[0298] Step F Synthesis of 2-methoxy-4-(5-((4-methylpiperazin-1-yl)methyl)isoxazol-3-yl)aniline (H7)

[0299] Add 6g 3-(3-methoxy-4-nitrophenyl)-5-((4-methylpiperazin-1-yl)methyl)isoxazole and 0.6g palladium on carbon (Pd / C) to 50mL methanol, introduce hydrogen, evacuate, and react at room temperature for 6h. Filter by suction, wash the filter cake with methanol (30mL×3), discard the filter cake, and retain the filtrate. Remove the filtrate solvent to obtain 5.08g of black oil, with a yield of 92.6%, MS (ESI) m / z: 303.2[M+H] +1 .

[0300] Step G (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((4-methylpiperazin-1-yl)methyl)isoxazol-3-yl)phenyl)

[0301] Synthesis of (amino)-5-methyl-7,8-dihydropyridin-6(5H)-one

[0302] Under nitrogen protection, (0.19 g, 0.65 mmol) 2-methoxy-4-(4-(((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)aniline, (0.23 g, 0.78 mmol) intermediate M6, (0.03 g, 0.03 mmol) Pd2(dba)3, (0.046 g, 0.15 mmol) X-Phos, (0.84 g, 2.6 mmol) cesium carbonate were added to 10 mL of tert-butanol and reacted at 90 ° C for 12 hours. The reaction solution was filtered with diatomaceous earth pad, the filtrate was evaporated to dryness, and purified by column chromatography to obtain 0.1 g of light yellow solid with a yield of 28%. MS (ESI) m / z: 561.3 [M+H] +

[0303] According to the synthesis method of Example 65, the compounds of Examples 66 and 67 were synthesized using the corresponding small molecule amines. The structures, chemical names, melting points, MS, 1 The H-NMR data are shown in Table 9.

[0304] Table 9 Example 65 to Example 67

[0305]

[0306] Example 68: (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(3-(4-methylpiperazine-1-carbonyl)-1,2,4-oxadiazol-5-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0307] Step A Preparation of (R)-3-(4-((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-3-methoxyphenyl)-1,2,4-oxadiazole-5-carboxylic acid ethyl ester (I1)

[0308] Dissolve 4.39 g of intermediate B1 in 20 mL of THF, add 2.6 mL of DIPEA, drop 1.3 mL of ethyl oxalyl chloride at 0 ° C, heat the reaction mixture to room temperature and keep reflux for 2 hours, cool the reaction mixture to room temperature and add 2N HCl aqueous solution (10 mL) and water (100 mL). Extract the product with ethyl acetate (3x 25 mL). Wash the organic layer with NaHCO3 aqueous solution and then with water. Dry the organic layer with anhydrous MgSO4 and filter. After rotary evaporation to remove the solvent, column chromatography gave 3.64 g of purple solid, with a yield of 70%, MS (ESI) m / z: 522.2 [M+H] + .

[0309] Step B Preparation of (R)-3-(4-((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-3-methoxyphenyl)-1,2,4-oxadiazole-5-carboxylic acid (I2)

[0310] 3.6 g of intermediate I was added to 20 mL of 10% NaOH aqueous solution, reacted at room temperature for 1.5 h, 1 M hydrochloric acid was added to adjust Ph = 2, filtered, the filter cake was washed with water and dried to obtain 1.75 g of white solid, yield 53%, MS (ESI) m / z: 494.2 [M+H] + .

[0311] Step C Preparation of (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(3-(4-methylpiperazine-1-carbonyl)-1,2,4-oxadiazol-5-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0312] At room temperature, 0.2 g of intermediate I2 was dissolved in 3 mL of dry DMF, HATU was added, and the mixture was stirred for 10 min. 0.05 g of N-methylpiperazine was added, and the mixture was stirred for 0.5 h. 0.1 g of DIPEA was added, and the reaction was continued for 3 h. 30 mL of water was added, and DCM was extracted and evaporated to dryness. Column chromatography was performed to obtain 0.18 g of a light yellow solid with a yield of 72%. MS (ESI) m / z: 576.3 [M+H] + .

[0313] According to the synthesis method of Example 68, the compounds of Examples 69 to 70 were synthesized using the corresponding small molecule amines. The structures, chemical names, melting points, MS,1 The H-NMR data are shown in Table 10.

[0314] Table 10 Example 69 to Example 70

[0315]

[0316] Example 71: (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)-1,3,4-oxadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0317] Step A Preparation of ((R)-2-(2-(4-((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-3-methoxybenzoyl)hydrazino)-2-oxoacetic acid ethyl ester (J1) 3.34 g of intermediate D4 was dissolved in 20 mL of DCM, 0.77 g of triethylamine was added, 1.03 g of ethyl oxalyl chloride was added under stirring at 0°C, and the reaction was continued at 0°C for 1 h, and then an appropriate amount of water was added. The organic phase was separated and evaporated to dryness to obtain 3 g of a white solid with a yield of 77%. MS (ESI) m / z: 540.2 [M+H] + Step B Preparation of (R)-5-(4-((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridin-2-yl)amino)-3-methoxyphenyl)-1,3,4-oxadiazole-2-carboxylic acid ethyl ester (J2)

[0318] 3 g of intermediate J1 was dissolved in 30 mL of phosphorus oxychloride, heated to 100 °C and reacted for 10 h. After evaporating the reaction solution, saturated sodium carbonate aqueous solution was added to adjust to neutrality, filtered, and the filter cake was washed with water and dried to obtain 1.78 g of gray solid, with a yield of 63%. MS (ESI) m / z: 522.2 [M+H] + .

[0319] Step C Preparation of (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)-1,3,4-oxadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0320] Using intermediate J2 as raw material, the synthetic method of step B and step C in Example 68 was used to obtain MS (ESI) m / z: 576.3 [M+H] + .

[0321] According to the synthesis method of Example 71, the compounds of Examples 72 to 73 were synthesized using the corresponding small molecule amines. The structures, chemical names, melting points, MS,1 H-NMR data are shown in Table 11.

[0322] Table 11 Example 71 to Example 73

[0323]

[0324] Example 74 (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)-1,3,4-thiadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0325] Step A Preparation of ethyl 2-(2-(3-methoxy-4-nitrobenzoyl)hydrazine)-2-oxoacetate (K1)

[0326] The intermediate E1 was used as the starting material and prepared according to the method of step A of Example 71. MS (ESI) m / z: 312.1 [M+H] + .

[0327] Step B Preparation of ethyl 5-(3-methoxy-4-nitrophenyl)-1,3,4-thiadiazole-2-carboxylate (K2)

[0328] The intermediate K1 was used as the raw material and prepared according to the method of step C of Example 51. MS (ESI) m / z: 310.1 [M+H] + .

[0329] Step C Preparation of 5-(3-methoxy-4-nitrophenyl)-1,3,4-thiadiazole-2-carboxylic acid (K3)

[0330] The intermediate K2 was used as the starting material and prepared according to the method of step B of Example 68. MS (ESI) m / z: 281.9 [M+H] + .

[0331] Step D Preparation of (5-(3-methoxy-4-nitrophenyl)-1,3,4-thiadiazol-2-yl)(4-methylpiperazine-1-yl)methanone (K4)

[0332] The intermediate K3 was used as the starting material and prepared according to the method of step C of Example 68. MS (ESI) m / z: 364.1 [M+H] + .

[0333] Step E Preparation of (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)-1,3,4-thiadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0334] The intermediate K4 was used as the raw material and prepared according to the method of step E and step F of Example 51. MS (ESI) m / z: 592.3 [M+H] + .

[0335] According to the synthesis method of Example 74, the compounds of Examples 75 to 76 were synthesized using the corresponding small molecule amines. The structures, chemical names, melting points, MS, 1 H-NMR data are shown in Table 12.

[0336] Table 12 Example 74 to Example 76

[0337]

[0338] Example 77 (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(4-(4-methylpiperazine-1-carbonyl)-1H-1,2,3-triazol-1-yl)phenyl))amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0339] Step A Preparation of 1-(3-methoxy-4-nitrophenyl)-1H-1,2,3-triazole-4-carboxylic acid (L1)

[0340] Propylic acid and intermediate C2 were used as raw materials and prepared according to the method of step B of Example 17. MS (ESI) m / z: 265.1 [M+H] + .

[0341] Step B Preparation of (1-(3-methoxy-4-nitrophenyl)-1H-1,2,3-triazol-4-yl)(4-methylpiperazin-1-yl)methanone (L2)

[0342] The intermediate L1 was used as the raw material and prepared according to the method of step C of Example 68. MS (ESI) m / z: 347.1 [M+H] + .

[0343] Step C Preparation of (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(4-(4-methylpiperazine-1-carbonyl)-1H-1,2,3-triazol-1-yl)phenyl))amino)-5-methyl-7,8-dihydropteridin-6(5H)-one

[0344] The intermediate L2 was used as the raw material and prepared according to the method of step E and step F of Example 17. MS (ESI) m / z: 575.3 [M+H] + .

[0345] According to the synthesis method of Example 77, the compounds of Examples 78 to 79 were synthesized using the corresponding small molecule amines. The structures, chemical names, melting points, MS, 1 H-NMR data are shown in Table 13.

[0346] Table 13 Example 77 to Example 79

[0347]

[0348] Example 80 (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)isoxazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropyridin-6(5H)-one

[0349] Step A Preparation of 3-(3-methoxy-4-nitrophenyl)isoxazole-5-carboxylic acid (M1)

[0350] Propylic acid and intermediate H3 were used as raw materials and prepared according to the method of step C of Example 65. MS (ESI) m / z: 265.1 [M+H] + .

[0351] Step B Preparation of (3-(3-methoxy-4-nitrophenyl)isoxazol-5-yl)(4-methylpiperazin-1-yl)methanone (M2)

[0352] The intermediate M1 and N-methylpiperazine were used as raw materials and prepared according to the method of step C of Example 68. MS (ESI) m / z: 347.1 [M+H ]+ .

[0353] Step C Preparation of (R)-8-cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)isoxazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropyridin-6(5H)-one

[0354] The intermediate M2 was used as the raw material and prepared according to the method of step F and step G of Example 65. MS (ESI) m / z: 575.3 [M+H] + .

[0355] According to the synthesis method of Example 80, the compounds of Examples 81 to 82 were synthesized using the corresponding small molecule amines. The structures, chemical names, melting points, MS, 1 H-NMR data are shown in Table 14.

[0356] Table 14 Example 80 to Example 82

[0357]

[0358]

[0359] The compounds of the present invention were studied for PLK1 enzyme activity, and the results are as follows

[0360] The in vitro PLK1 enzyme activity test was performed on the dihydropteridone compound containing a benzobiheteroaryl group represented by the above formula I of the present invention using a fluorescence analysis method. The compounds described below are all dihydropteridone compounds containing a benzobiheteroaryl group represented by the above formula I prepared in the above examples.

[0361] Dilute PLK1 kinase to 20 ng / μL in 5x KB DTT-MgCl2 assay buffer. Add 4 μL of inhibitor solution to each test well, while adding 4 μL of the same solution without inhibitor (inhibitor buffer) to the wells labeled "Negative Control" and "Positive Control". Add 2 μL of STK Substrate-biotin to all wells. Add 2 μL of kinase to the test wells and 2 μL of 5x KB DTT-MgCl2 assay buffer to the wells labeled "Negative Control" and "Positive Control". Add 2 μL of ATP to all wells. Seal and incubate at room temperature for 1 hour.

[0362] Add 5 μL of Sa-XL665 and STK Ab-Cryptate to all wells in sequence, seal and incubate at room temperature for 1 h.

[0363] The fluorescence signal was detected using SpectraMax i3X multi-function microplate reader: Emission Ratio (ER) = 665 nm Emission signal / 615 nm Emission signal (320 nm excitation, 665 nm, 615 nm emission).

[0364] The inhibition rate was calculated according to the formula: Inhibition rate = (ER positive -ER sample ) / (ER positive -ER negative )*100%(ER positive : ER value of positive control well; ER sample : ER value of the test hole; ER negative : ER value of negative control well).

[0365] Finally, SPSS software was used to input the data and draw the inhibition rate-concentration curve to calculate the IC 50 .

[0366] Table 15 PLK1 enzyme IC of some examples 50 Activity data

[0367] Example <![CDATA[IC 50 (nM)]]> Example 1 0.37 Example 2 0.66 Example 3 0.71 Example 6 0.68 Example 7 0.59 Example 8 0.45 Example 9 0.91 Example 10 0.35 Embodiment 11 0.26 Example 12 0.18 Embodiment 13 0.37 Embodiment 17 0.42 Embodiment 18 0.49 Embodiment 19 0.44 Embodiment 20 0.31 Embodiment 26 0.23 Embodiment 27 0.28 Embodiment 30 0.64 Embodiment 31 0.44 Embodiment 34 0.37 Embodiment 38 0.23 Embodiment 46 0.12 Embodiment 51 0.29 Embodiment 52 0.35 Embodiment 53 0.27 Embodiment 54 0.39 Embodiment 55 0.13 Embodiment 56 0.45 Embodiment 61 0.52 Embodiment 65 0.33 Embodiment 68 0.61 Embodiment 70 0.77 Embodiment 74 0.22 Embodiment 77 0.36 Embodiment 80 0.53 BI6727 0.82

[0368] The compounds of the general formula recorded in the above table all have good PLK1 inhibitory activity, and most of the compounds are equivalent to or better than the positive control drug BI 6727.

[0369] Then, the dihydropteridone derivatives containing phenylheteroaryl groups according to the above formula I of the present invention were further subjected to in vitro experiments on the inhibition of the activities of human breast cancer cells MDA-MB-231 and human leukemia cells MV4-11.

[0370] CCK-8 assay to detect the antiproliferative activity of the compounds (MDA-MB-231 and MV4-11 cell lines)

[0371] Cells in the logarithmic growth phase were cultured at a rate of 8 to 15 × 10 4 / mL was inoculated into a 96-well plate, and the cell solution volume was 100μL / well. 100μL of different concentrations of drug solution was added to each well, and three replicates were set for each concentration. Incubate for 48h under standard conditions. After the incubation, the 96-well plate was removed, 20μL of CCK-8 solution was added to each well, and the incubation continued for 4h before being removed and placed on a micro-oscillator for 5min. The absorbance (OD) of each well was measured at 450nm on an ELISA reader.

[0372] The inhibition rate (IR) of cell proliferation in vitro was calculated according to the formula:

[0373] IR%=(OD control -OD sample ) / (OD control -OD blank )×100%

[0374] In the formula, OD control : OD value of the well without drug addition; OD sample : OD value of the dosing hole; OD blank : In the CCK-8 method, it is the OD value of the wells with only culture medium added; in the MTT method, it is the OD value of the wells with only DMSO added.

[0375] Finally, SPSS software was used to input the data and draw the inhibition rate-concentration curve to calculate the IC 50 .

[0376] The test results of some compounds in MDA-MB-231 and MV4-11 cell lines are shown in Table 16.

[0377] Table 16 Cell IC of some examples 50 Activity data

[0378]

[0379]

[0380] The results of the cell activity test of the compounds showed that the activities of most compounds against MDA-MB-231 and MV4-11 were equivalent to or better than the positive control drug BI 6727.

[0381] At the same time, the results of in vitro kinase and cell activity tests show that the compounds involved in the present invention have good PLK1 enzyme activity and excellent cell activity, and the activity of some compounds is better than that of the positive control drug PLK1.

Claims

1. A dihydropteridone derivative containing a phenylheteroaryl group, characterized in that: The derivatives are compounds represented by the general formula I and their salts, I in, R1 is (C1-C6) alkyl, (C3-C6) cycloalkyl; R2 is (C1-C6) alkyl, (C3-C6) cycloalkyl, 3-6 membered heterocyclic group, (C6-C 12 )spirocyclic group, 6-12 membered heterospirocyclic group, (C6-C 10 ) aryl, 5-10 membered heteroaryl, the alkyl, cycloalkyl, heterocyclyl, spirocyclyl, heterospirocyclyl, aryl or heteroaryl is optionally substituted by 1-3 identical or different R6; R6 is hydrogen, halogen, hydroxy, cyano, amino, nitro, (C1-C6) alkyl, (C1-C6) haloalkyl, (C1-C6) alkoxy; R3 is hydrogen, (C1-C6)alkoxy, (C3-C6)cycloalkoxy which is unsubstituted or substituted by at least one of the following groups; the following groups are halogen, hydroxyl or amino; Ring A is selected from (C6-C 10 ) aryl, 5-6 membered heteroaryl, said aryl or heteroaryl being optionally substituted by 1-3 identical or different R9; R9 is hydrogen, halogen, hydroxy, cyano, amino, nitro, (C1-C6)haloalkyl, (C1-C6)alkyl, (C1-C6)alkoxy; R4 and R5 are the same or different and are independently selected from hydrogen, (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl, (C2-C6) alkenyl, (C2-C6) alkynyl, dimethylaminoethyl, and the alkyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 identical or different R7; or R4 and R5 together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclic group containing at least one heteroatom; the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, the heterocyclic group is optionally substituted with one or more oxo groups, or the heterocyclic group is optionally substituted with 0-3 identical or different R8 groups; R7 and R8 are the same or different and are independently selected from hydrogen, hydroxy, halogen, amino substituted with 0-2 (C1-C6) alkyl groups, cyano, (C1-C6) alkyl acyl, (C1-C6) alkyl sulfonyl, (C1-C6) alkyl which is unsubstituted or substituted with at least one of the following groups, (C1-C6) alkoxy, (C3-C6) cycloalkyl, (C3-C6) alkylsulfonyl, 10 )cycloalkyl-(C1-C4)alkyl; the following groups are hydroxy, amino or halogen; X is an oxo-substituted or unoxo-substituted (C1-C6) alkyl group; m is an integer from 0 to 1; The above heterocyclic group or heteroaryl group may contain 1 to 4 heteroatoms, which may be the same or different and are selected from N, O and S.

2. The dihydropteridone derivative containing a phenyl biheteroaryl group according to claim 1, characterized in that: In the general formula I, R1 is (C1-C4) alkyl, (C3-C5) cycloalkyl; R2 is (C1-C6) alkyl, (C3-C6) cycloalkyl, 3-6 membered heterocyclic group, (C6-C 10 ) aryl, 5-6 membered heteroaryl, the alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl group is optionally substituted by 1-3 identical or different R6; R6 is hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethyl, (C1-C6) alkyl, (C1-C6) alkoxy; R3 is unsubstituted or substituted by at least one of the following groups (C1-C3) alkoxy; the following groups are halogen, hydroxyl or amino; Ring A is selected from phenyl, 5-6 membered heteroaryl, and the phenyl or heteroaryl is optionally substituted by 1-3 identical or different R9; R9 is hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethyl, (C1-C3) alkyl, (C1-C3) alkoxy; R4 and R5 are the same or different and are independently selected from hydrogen, (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl, dimethylaminoethyl, wherein the alkyl, cycloalkyl, heterocycloalkyl are optionally substituted by 1-3 identical or different R7; or R4 and R5 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing at least one heteroatom; the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, the heterocyclic group is optionally substituted with one or more oxo groups, or the heterocyclic group is optionally substituted with 0-3 identical or different R8 groups; R7 and R8 are the same or different and are independently selected from hydrogen, hydroxyl, halogen, amino substituted with 0-2 (C1-C4) alkyl groups, cyano, (C1-C4) alkylacyl, (C1-C4) alkylsulfonyl, (C1-C4) alkyl unsubstituted or substituted with at least one of the following groups, (C1-C4) alkoxy, (C3-C6) cycloalkyl, (C3-C6) cycloalkyl-(C1-C4) alkyl; the following groups are hydroxyl, amino or halogen; X is an oxo-substituted or unoxo-substituted (C1-C4) alkyl group; m is an integer from 0 to 1; The above heterocyclic group or heteroaryl group may contain 1 to 4 heteroatoms, which may be the same or different and are selected from N, O and S.

3. The dihydropteridone derivative containing a phenyl biheteroaryl group according to claim 2, characterized in that: In the general formula I, R1 is (C1-C4) alkyl; R2 is (C1-C6) alkyl, (C3-C6) cycloalkyl, 3-6 membered heterocyclic group, (C6-C 10 ) aryl, 5-6 membered heteroaryl, the alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl group is optionally substituted by 1-3 identical or different R6; R6 is hydrogen, halogen, hydroxy, cyano, amino, nitro, trifluoromethyl, (C1-C6) alkyl, (C1-C6) alkoxy; R3 is (C1-C3) alkoxy; Ring A is selected from , , , , , , , ; R4 and R5 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl, (C3-C5) cycloalkyl, (C3-C6) heterocycloalkyl, and the alkyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 identical or different R7; or R4 and R5 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing at least one heteroatom; the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, the heterocyclic group is optionally substituted with one or more oxo groups, or the heterocyclic group is optionally substituted with 0-3 identical or different R8 groups; R7 and R8 are the same or different and are independently selected from hydrogen, hydroxyl, halogen, amino substituted with 0-2 (C1-C6) alkyl groups, cyano, (C1-C3) alkylacyl, (C1-C3) alkylsulfonyl, (C1-C3) alkyl unsubstituted or substituted with at least one of the following groups, (C1-C3) alkoxy, (C3-C6) cycloalkyl, (C3-C6) cycloalkyl-(C1-C4) alkyl; the following groups are hydroxyl, amino or halogen; X is an oxo-substituted or unoxo-substituted (C1-C4) alkyl group; m is an integer from 0 to 1; The above heterocyclic group or heteroaryl group may contain 1 to 4 heteroatoms, which may be the same or different and are selected from N, O and S.

4. The dihydropteridone derivative containing a phenylbiheteroaryl group according to claim 3, characterized in that: In the general formula I, R1 is methyl R2 is (C3-C6) alkyl, (C3-C6) cycloalkyl, 3-6 membered heterocyclyl, phenyl, thienyl, furyl, naphthyl, wherein the alkyl, cycloalkyl, heterocyclyl, phenyl, thienyl, furyl, naphthyl is optionally substituted by 1-3 identical or different R6; R6 is hydrogen, halogen, (C1-C4) alkyl, (C1-C4) alkoxy, trifluoromethyl, hydroxyl, cyano; R3 is (C1-C3) alkoxy; Ring A is selected from , , , , , , , ; R4 and R5 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl, (C3-C5) cycloalkyl, (C3-C6) heterocycloalkyl, and the alkyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 identical or different R7; or R4 and R5 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing at least one heteroatom; the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, the heterocyclic group is optionally substituted with one or more oxo groups, or the heterocyclic group is optionally substituted with 0-3 identical or different R8 groups; R7 and R8 are the same or different and are independently selected from hydrogen, hydroxyl, halogen, amino substituted with 0-2 (C1-C6) alkyl groups, cyano, (C1-C3) alkylacyl, (C1-C3) alkylsulfonyl, (C1-C3) alkyl unsubstituted or substituted with at least one of the following groups, (C1-C3) alkoxy, (C3-C6) cycloalkyl, (C3-C6) cycloalkyl-(C1-C4) alkyl; the following groups are hydroxyl, amino or halogen; X is an oxo-substituted or unoxo-substituted (C1-C4) alkyl group; m is an integer from 0 to 1; The above heterocyclic group or heteroaryl group may contain 1 to 4 heteroatoms, which may be the same or different and are selected from N, O and S.

5. The dihydropteridone derivative containing a phenylbiheteroaryl group according to claim 4, characterized in that: In the general formula I, R1 is methyl R2 is (C3-C6) alkyl, (C3-C6) cycloalkyl, 3-6 membered heterocyclyl, phenyl, thienyl, furyl, naphthyl, wherein the alkyl, cycloalkyl, phenyl, heterocyclyl, thienyl, furyl, naphthyl is optionally substituted by 1-3 identical or different R6; R6 is hydrogen, halogen, (C1-C4) alkyl, (C1-C4) alkoxy, trifluoromethyl, hydroxyl, cyano; R3 is methoxy; Ring A is selected from , , , , , , , ; R4 and R5 are the same or different and are independently selected from hydrogen, (C1-C6) alkyl, (C3-C5) cycloalkyl, (C3-C6) heterocycloalkyl, and the alkyl, cycloalkyl or heterocycloalkyl is optionally substituted by 1-3 identical or different R7; or R4 and R5 together with the nitrogen atom to which they are attached form , , , , , , , , , , , , , , , , , , , , , , , , , , , or ; X is CH2 or CO; m is an integer from 0 to 1; The above heterocyclic group or heteroaryl group may contain 1 to 4 heteroatoms, which may be the same or different and are selected from N, O and S.

6. The dihydropteridone derivative containing a phenyl biheteroaryl group according to claim 5, characterized in that: The derivative is a compound of formula I or a pharmaceutically acceptable salt thereof, (R)-8-Cyclopentyl-2-((4-(4-((dimethylamino)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-2-((4-(4-((4-cyclopentylpiperazin-1-yl)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-2-((4-(4-((diethylamino)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((4-(4-((4-ethylpiperazin-1-yl)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-2-((4-(4-((cyclopropylamino)methyl)thiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(piperazin-1-ylmethyl)-1,2,4-oxadiazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((4-methylpiperazin-1-yl)methyl)-1,2,4-oxadiazol-3-yl))phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-2-((4-(5-((4-cyclopentylpiperazin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((4-(5-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-1,2,4-oxadiazol-3-yl))-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((4-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-2-((4-(5-(((3S,5R)-3,5-dimethylpiperazin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(4-((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((4-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-2-((4-(4-(((3S,5R)-3,5-dimethylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((4-(4-((4-isopropylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-2-((4-(5-((dimethylamino)methyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((3-methylpiperazin-1-yl)methyl)-1,3,4-oxadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-2-((4-(5-(((2-(dimethylamino)ethyl)(methyl)amino)methyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((4-(5-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-1,3,4-oxadiazol-2-yl))-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-cyclopentyl-2-((4-(5-((3-(dimethylamino)pyrrolidin-1-yl)methyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((4-(5-(((R)-2-(hydroxymethyl)pyrrolidin-1-yl)methyl)-1,3,4-oxadiazol-2-yl)-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-cyclohexyl-7-ethyl-2-((4-(5-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-1,3,4-oxadiazol-2-yl))-2-methoxyphenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((3-methylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-2-((4-(5-(((3S,5R)-3,5-dimethylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((4-methylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl))phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(piperazin-1-ylmethyl)-1,3,4-thiadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-2-((4-(5-((4-acetylpiperazin-1-yl)methyl)-1,3,4-thiadiazol-2-yl)-2-methoxyphenyl)amino)-8-cyclopentyl-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(piperazin-1-ylmethyl)oxazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-2-((4-(2-((dimethylamino)methyl)-1H-pyrrol-1-yl)-2-methoxyphenyl)amino)-7-ethyl-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-((4-methylpiperazin-1-yl)methyl)isoxazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropyridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(3-(4-methylpiperazine-1-carbonyl)-1,2,4-oxadiazol-5-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)-1,3,4-oxadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)-1,3,4-thiadiazol-2-yl)phenyl)amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(4-(4-methylpiperazine-1-carbonyl)-1H-1,2,3-triazol-1-yl)phenyl))amino)-5-methyl-7,8-dihydropteridin-6(5H)-one (R)-8-Cyclopentyl-7-ethyl-2-((2-methoxy-4-(5-(4-methylpiperazine-1-carbonyl)isoxazol-3-yl)phenyl)amino)-5-methyl-7,8-dihydropyridin-6(5H)-one.

7. The dihydropteridone derivative containing a phenyl biheteroaryl group according to any one of claims 1 to 6, characterized in that: The compound salt is a salt formed with an acid, and the acid is selected from: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, trifluoroacetic acid or aspartic acid.

8. A pharmaceutical composition, characterized in that: The composition contains the compound of general formula I according to any one of claims 1 to 6 and a pharmaceutically acceptable salt thereof.

9. Use of the compound according to any one of claims 1 to 6 and a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or treating a disease associated with a PLK1 kinase inhibitor.

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