2-aminopyrimidine-5-carboxamide compound and application thereof in preparing medicine for treating and / or preventing cancer

Abstract

The invention relates to the field of medicinal chemistry, in particular to a 2-aminopyrimidine-5-formamide compound and application thereof in preparation of drugs for treating and / or preventing cancers. The compound is a 2-aminopyrimidine-5-formamide compound represented by a general formula I, and pharmaceutically acceptable salts, solvates or prodrugs thereof, wherein substituent groups R1, R2 and R3 have meanings given in the specification. The invention further relates to application of the compound shown in the general formula I in preparation of drugs for treating and / or preventing diseases related to Wee1 kinase. # imgabs0 #

Description

Technical field:

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to a 2-aminopyrimidine-5-carboxamide compound and its use in preparing a drug for treating and / or preventing cancer. Background technology:

[0002] Malignant tumors are diseases involving abnormal cell growth with the potential to invade or spread to other parts of the body. Finding effective tumor treatments has always been a research focus in the field of medicine in my country and even in the world. With the rapid development of molecular biology and tumor pathology research, molecular targeted therapy targeting tumor growth mechanisms has become an important strategy for tumor treatment. Protein tyrosine kinases are currently the most intensively studied. In recent years, countless small molecule tyrosine kinase inhibitors (TKIs) have been launched on the market, but research on serine / threonine protein kinases is very limited. Therefore, rational research on this kinase and the development of inhibition of this kinase are of great significance for cancer treatment.

[0003] Wee1 protein kinase belongs to the serine / threonine family, whose family members also include Wee1A, Wee1B and Myt1, and plays an important role in the process of cell mitosis. This protein kinase includes three domains: N-terminal regulatory domain, central kinase domain and C-terminal regulatory domain. Wee1 plays a key regulatory role in the mitotic G2 / M checkpoint. When DNA is damaged, Wee1 inhibits CDK1 activity by phosphorylating the Tyr14 / 15 site of CDK1, inactivates the CDK1 / cyclinB complex, and prevents cells from entering mitosis, allowing sufficient time for damaged DNA to be repaired. However, inhibiting Wee1 will force immature cells or cells with DNA damage to enter mitosis prematurely, thereby triggering mitotic catastrophe and ultimately leading to cell apoptosis.

[0004] Synthetic lethality refers to the phenomenon that two or more non-lethal genes are simultaneously inactivated, leading to cell death. In normal cells, DNA damage can also occur through two pathways mediated by the tumor suppressor gene p53: ATM / ATR-P53-CDK4 / CyclinD and ATM / ATR-P53-CDK2 / CyclinE. However, in tumor cells, the mutation rate of the tumor suppressor gene p53 is high, with mutations in this gene present in more than 50% of tumor cells. The G1 / S checkpoint of tumor cells with defective p53 function is inactivated, which makes DNA repair mainly dependent on the G2 / M checkpoint. Wee1 inhibitors can drive cells to enter the division phase prematurely, thereby producing a synthetic lethality effect. Theoretically, inhibiting the activity of Wee1 kinase can selectively kill p53-deficient tumor cells without affecting normal cells. Related studies have shown that Wee1 kinase is overexpressed in liver cancer, lung cancer, colorectal cancer, gastric cancer, breast cancer, ovarian cancer, cervical cancer, squamous cell carcinoma, malignant melanoma, glioblastoma, and intracranial diffuse glioma, and its high expression in ovarian cancer, melanoma and glioblastoma is associated with poor prognosis. Therefore, inhibiting Wee1 kinase is expected to become a new strategy for treating cancer.

[0005] At present, many small molecule inhibitors of Wee1 have been reported, which are mainly divided into: pyrrolocarbazoles, such as PD0407824; pyrazolopyrimidinones, such as AZD1775, CJM061 and SC0191; dichlorophenyls, such as PD0166285 and IMP7068 (the structural formula is shown below). Among the above small molecule inhibitors, only AZD1775 has entered clinical research, and the rest are in preclinical research. AZD1775 is currently in the Phase II clinical research stage, but it has obvious drug tolerance problems. Therefore, intermittent administration in clinical trials has weakened its efficacy. The drug resistance problem of Wee1 small molecule inhibitors may be due to their poor selectivity for kinases. Therefore, it is crucial to study and develop a new highly selective inhibitor that can be continuously administered in clinical trials to achieve good clinical efficacy.

[0006] Summary of the invention:

[0007] The present invention aims at a 2-aminopyrimidine-5-carboxamide compound and its use in preparing a drug for treating and / or preventing cancer.

[0008] To achieve the above purpose, the present invention adopts the following technical solution:

[0009] A 2-aminopyrimidine-5-carboxamide compound, which is a 2-aminopyrimidine-5-carboxamide compound of general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof,

[0010]

[0011] in,

[0012] R 1 is unsubstituted or substituted with at least one identical or different R 4 Substituted (C6-C 10 ) aryl, 5-10 membered heteroaryl;

[0013] R 4 is hydroxy, halogen, nitro, amino, cyano, (C1-C6)alkyl or (C1-C6)alkoxy which is unsubstituted or substituted by at least one of the following substituents which are the same or different, wherein the following substituents are hydroxy, amino or halogen;

[0014] R 2 is (C1-C6) alkyl or (C3-C7) cycloalkyl, wherein the alkyl includes a straight chain or a branched chain alkyl;

[0015] R 3 is unsubstituted or substituted with 1 to 3 identical or different R 5 Substituted (C6-C 10 ) aryl, 5-10 membered heteroaryl, tetrahydroquinolyl, tetrahydroisoquinolyl;

[0016] R 5 is (C1-C6) haloalkyl, hydroxy, halogen, cyano, (C1-C6) alkyl, (C1-C6) alkoxy, -NR 6 R 7 、-(CH2) p NR 6 R 7 、-O(CH2) p NR 6 R 7 、-NH(CH2) p NR 6 R 7 、-NCH3(CH2) p NR 6 R 7 , a 5-10 membered heterocyclic group, the heterocyclic group optionally containing 1-3 double bonds and / or optionally 1-3 identical or different R 8 Substituted, wherein the heteroatoms in the heterocyclic group are 1 to 4 identical or different N, O and / or S;

[0017] Among them, R 6 and R 7 are the same or different, each independently selected from hydrogen, unsubstituted or substituted with 1-3 identical or different R 9 Substituted (C1-C6) alkyl, (C3-C7) cycloalkyl, (C6-C 10) aryl, 5-10 membered heterocyclic group;

[0018] or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 5-10 membered heterocyclic group, the heterocyclic group optionally containing 1-3 double bonds and / or optionally substituted by 1-3 identical or different R 10 Substituted, wherein the heteroatoms in the heterocyclic group are 1 to 4 identical or different N, O and / or S;

[0019] R 8 , R 9 and R 10 are the same or different and are independently selected from oxo, hydroxyl, halogen, amino, (C1-C6) alkylacyl, (C1-C6) alkoxyformyl, (C1-C6) alkylsulfonyl, (C5-C 10 ) heterocyclic group, free, salified, esterified or amidated carboxyl group, (C1-C6) alkyl group which is unsubstituted or substituted by at least one hydroxyl group, amino group or halogen group, (C1-C6) alkoxy group, (C3-C7) cycloalkyl group;

[0020] p is an integer from 1 to 4.

[0021] Preferably, the compound is a 2-aminopyrimidine-5-carboxamide compound of general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein:

[0022] R 1 is unsubstituted or substituted with 1 to 3 identical or different R 4 Substituted phenyl;

[0023] R 4 is hydroxy, halogen, nitro, amino, cyano, (C1-C4)alkyl or (C1-C4)alkoxy which is unsubstituted or substituted by at least one of the following substituents which are the same or different, wherein the following substituents are hydroxy, amino or halogen;

[0024] R 2 is (C1-C4) alkyl or (C3-C6) cycloalkyl, wherein the alkyl includes a straight chain or a branched chain alkyl;

[0025] R 3 is unsubstituted or substituted with 1 to 3 identical or different R 5 Substituted phenyl, 5-6 membered heteroaryl, tetrahydroquinolyl, tetrahydroisoquinolyl;

[0026] R 5 is trifluoromethyl, hydroxyl, halogen, cyano, (C1-C4) alkyl, (C1-C4) alkoxy, -NR 6 R 7 、-(CH2) pNR 6 R 7 、-O(CH2) p NR 6 R 7 、-NH(CH2) p NR 6 R 7 、-NCH3(CH2) p NR 6 R 7 , a 5-6 membered heterocyclic group, the heterocyclic group optionally containing 1-2 double bonds and / or optionally 1-3 identical or different R 8 Substituted, wherein the heteroatoms in the heterocyclic group are 1 to 3 identical or different N, O and / or S;

[0027] Among them, R 6 and R 7 are the same or different, each independently selected from hydrogen, unsubstituted or substituted with 1-3 identical or different R 9 Substituted (C1-C4) alkyl, (C3-C6) cycloalkyl, phenyl, 5-6 membered heterocyclic group;

[0028] or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 5-10 membered heterocyclic group, the heterocyclic group optionally containing 1-2 double bonds and / or optionally substituted by 1-3 identical or different R 10 Substituted, wherein the heteroatoms in the heterocyclic group are 1 to 3 identical or different N, O and / or S;

[0029] R 8 , R 9 and R 10 are the same or different and are independently selected from oxo, hydroxyl, halogen, amino, (C1-C4) alkylacyl, (C1-C4) alkoxyformyl, (C1-C4) alkylsulfonyl, (C5-C6) heterocyclyl, free, salified, esterified or amidated carboxyl, (C1-C4) alkyl, (C1-C4) alkoxy, (C3-C6) cycloalkyl which is unsubstituted or substituted with at least one hydroxyl, amino or halogen;

[0030] p is 1, 2 or 3.

[0031] Preferably, the compound is a 2-aminopyrimidine-5-carboxamide compound of general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein:

[0032] R 1 is unsubstituted or substituted with 1 to 3 identical or different R 4 Substituted phenyl,;

[0033] R4 is hydroxy, halogen, nitro, amino, cyano, (C1-C4)alkyl or (C1-C4)alkoxy which is unsubstituted or substituted by at least one of the following substituents which are the same or different, wherein the following substituents are hydroxy, amino or halogen;

[0034] R 2 is (C1-C4) alkyl or (C3-C4) cycloalkyl, wherein the alkyl includes a straight chain or a branched chain alkyl;

[0035] R 3 is unsubstituted or substituted with 1 to 3 identical or different R 5 substituted phenyl, pyridyl, pyrazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, tetrahydroquinolyl, tetrahydroisoquinolyl;

[0036] R 5 is trifluoromethyl, hydroxyl, halogen, cyano, (C1-C3) alkyl, (C1-C3) alkoxy, -NR 6 R 7 、-(CH2) p NR 6 R 7 、-O(CH2) p NR 6 R 7 、-NH(CH2) p NR 6 R 7 、-NCH3(CH2) p NR 6 R 7 , a 5-6 membered heterocyclic group, the heterocyclic group is optionally substituted by 1-2 identical or different R 8 Substitution; wherein the heteroatom in the heterocyclic group contains 1-2 identical or different N, O and / or S;

[0037] Among them, R 6 and R 7 are the same or different, each independently selected from hydrogen, unsubstituted or substituted with 1-3 identical or different R 9 Substituted (C1-C4) alkyl, (C3-C4) cycloalkyl, phenyl, 5-6 membered heterocyclic group;

[0038] or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 5-6 membered heterocyclic group, the heterocyclic group optionally containing 1-2 double bonds and / or optionally substituted by 1-3 identical or different R 10 Substituted, wherein the heteroatoms in the heterocyclic group are 1 to 3 identical or different N, O and / or S;

[0039] R 8 , R 9and R 10 are the same or different and are independently selected from oxo, hydroxyl, halogen, amino, (C1-C4) alkylacyl, (C1-C4) alkoxyformyl, (C1-C4) alkylsulfonyl, (C5-C6) heterocyclyl, free, salified, esterified or amidated carboxyl, (C1-C4) alkyl, (C1-C4) alkoxy, (C3-C6) cycloalkyl which is unsubstituted or substituted with at least one hydroxyl, amino or halogen;

[0040] p is 1, 2 or 3.

[0041] Still more preferably, the compound is a 2-aminopyrimidine-5-carboxamide compound of general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein:

[0042] R 1 is unsubstituted or substituted with 1 to 3 identical or different R 4 Substituted phenyl;

[0043] R 4 is hydroxy, halogen, nitro, amino, cyano, (C1-C4)alkyl or (C1-C4)alkoxy which is unsubstituted or substituted by at least one of the following substituents which are the same or different, wherein the following substituents are hydroxy, amino or halogen;

[0044] R 2 is methyl, ethyl, cyclopropyl, cyclobutyl;

[0045] R 3 is unsubstituted or substituted with 1 to 3 identical or different R 5 substituted phenyl, pyridyl, pyrazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, tetrahydroquinolyl, tetrahydroisoquinolyl;

[0046] R 5 For-NR 6 R 7 、-(CH2) p NR 6 R 7 、-O(CH2) p NR 6 R 7 、-NH(CH2) p NR 6 R 7 、-NCH3(CH2) p NR 6 R 7 ;

[0047] Among them, R 6 and R 7are the same or different and are independently selected from hydrogen, (C1-C4) alkyl, (C3-C4) cycloalkyl;

[0048] or R 6 and R 7 Together with the nitrogen atoms to which they are attached, they form

[0049] p is 1, 2 or 3.

[0050] More preferably, the 2-aminopyrimidine-5-carboxamide compounds of general formula I and pharmaceutically acceptable salts, solvates or prodrugs thereof:

[0051] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0052] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[4-(4-ethylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0053] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[4-(piperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[4-(3-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0054] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[3-methyl-4-(4-ethylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0055] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[3-methyl-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0056] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[3-methyl-4-(piperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0057] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[3-methoxy-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0058] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[3-fluoro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0059] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[2-fluoro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0060] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[3-(trifluoromethyl)-fluoro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0061] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[3-chloro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0062] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[3-cyano-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0063] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[6-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}pyrimidine-5-carboxamide

[0064] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[4-(dimethylamino)phenyl]amino}pyrimidine-5-carboxamide

[0065] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{{4-[2-(dimethylamino)ethyl]phenyl}amino}pyrimidine-5-carboxamide

[0066] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{{4-{[2-(dimethylamino)ethyl](methyl)amino}phenyl}amino}pyrimidine-5-carboxamide

[0067] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{{4-[2-(diethylamino)ethoxy]phenyl}amino}pyrimidine-5-carboxamide

[0068] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{{4-[2-(4-methylpiperazin-1-yl)ethoxy]phenyl}amino}pyrimidine-5-carboxamide

[0069] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{{4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino}pyrimidine-5-carboxamide

[0070] N-(2,6-Dichlorophenyl)-4-ethoxy-2-[(4-morpholinylphenyl)amino]pyrimidine-5-carboxamide

[0071] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[4-(4-acetylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0072] 4-{4-{{5-[(2,6-dichlorophenyl)carbamoyl]-4-ethoxypyrimidin-2-yl}amino}phenyl}piperazine-1-carboxylic acid ethyl ester

[0073] tert-Butyl 4-{4-{{5-[(2,6-dichlorophenyl)carbamoyl]-4-ethoxypyrimidin-2-yl}amino}phenyl}piperazine-1-carboxylate

[0074] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{{[4-(methylsulfonyl)piperazin-1-yl]phenyl}amino}pyrimidine-5-carboxamide

[0075] N-(2,6-Dichlorophenyl)-4-ethoxy-2-{[1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl]amino}pyrimidine-5-carboxamide

[0076] N-(2,6-Dichlorophenyl)-4-ethoxy-2-[(1,2,3,4-tetrahydroisoquinolin-7-yl)amino]pyrimidine-5-carboxamide

[0077] N-(2,6-Dichlorophenyl)-4-ethoxy-2-[(2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)amino]pyrimidine-5-carboxamide

[0078] According to some common methods in the field to which the present invention belongs, the compound of the general formula I of the present invention can form its pharmaceutically acceptable salt with acid. Preferred acids are 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 and aspartic acid.

[0079] A pharmaceutical composition comprising a compound of general formula I and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof.

[0080] An application of the compound, and use of the compound and its pharmaceutically acceptable salts, hydrates, solvates or prodrugs in the preparation of drugs for treating and / or preventing cancer.

[0081] Use of the compound and its pharmaceutically acceptable salt, hydrate, solvate or prodrug in preparing drugs for preventing or treating diseases associated with Wee1 kinase.

[0082] The diseases mediated by Wee1 are prevented from being breast cancer, lung cancer, cervical cancer, ovarian cancer, prostate cancer, melanoma and liver cancer.

[0083] Through the test of Wee1 enzyme activity, it was found that the compound of the present invention has significant inhibition of Wee1 kinase activity and has a strong inhibitory effect on NCI-H1299 lung cancer cells with high expression of Wee1.

[0084] The exact amount of the compound of the present invention required to treat breast and lung cancer disease will vary from subject to subject, depending on the species, 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 appropriate effective amount can be determined by one of ordinary skill in the art using only routine experimentation.

[0085] The compound can be administered in an amount of about 0.1 to 100 mg / kg body weight per day, preferably 1 to 50 mg / kg body weight per day. It is understood that the dosage may vary depending on the patient's needs, the severity of the proliferative disease or malignant blood disease 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.

[0086] Mammal means a human or an animal.

[0087] 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.

[0088] 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.

[0089] 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 does not limit the scope of the present invention in any way.

[0090] The following synthetic routes describe the preparation of the derivatives of formula I of the present invention, all starting materials being prepared by the methods described in these schemes, by methods well known to those skilled in the art of organic chemistry, or commercially available. All final compounds of the present invention are prepared by the methods described in these schemes or by methods analogous thereto, which are well known to those skilled in the art of organic chemistry. All variables used in these schemes are as defined below or as defined in the claims.

[0091] According to the compound of general formula I of the present invention, in route A, the following compound is taken as an example: the substituent R 1 , R 2 , R 4 , R 5 , R 6 and R 7 The definition is the same as that in the claims.

[0092] Route A:

[0093]

[0094] In route A, 4-chloro-2-methylthiopyrimidine-5-carboxylic acid ethyl ester is used as the starting material, and intermediate C is obtained by hydrolysis and chlorination reaction; then, intermediate C is reacted with R 1 -NH2 is subjected to an amide condensation reaction to obtain intermediate D; a substitution reaction is carried out under alkaline conditions to obtain intermediate E; using substituted p-fluoronitrobenzene as a raw material, intermediate H is obtained through substitution and reduction reactions; intermediate E is oxidized to sulfone by m-CPBA and then subjected to a substitution reaction with intermediate H to obtain compound Ii of general formula I.

[0095] When R 3 is a heteroaryl group (such as 1H-pyrazol-4-yl), compound I-ii of general formula I can be synthesized according to route B:

[0096] Route B:

[0097]

[0098] In route B, 4-nitro-1H-pyrazole is used as the starting material, and intermediate K is obtained through Mitsunobu reaction and reduction reaction. Intermediate E is oxidized to sulfone by m-CPBA and then undergoes substitution reaction with intermediate K to obtain compound I-ii of general formula I.

[0099] When R 3 When it is phenyl, compound I-iii of general formula I can be synthesized according to route C:

[0100] Route C:

[0101]

[0102] In route C, substituted p-nitrophenol is used as the starting material, and intermediate N is obtained through substitution and reduction reaction. Intermediate E is oxidized to sulfone by m-CPBA and then undergoes substitution reaction with intermediate N to obtain compound I-iii of general formula I.

[0103] When R 3 When it is a tetrahydroquinoline group, the compound I-iiii of the general formula I can be synthesized according to route C:

[0104] Route D:

[0105]

[0106] In route D, substituted 7-nitro-1,2,3,4-tetrahydroisoquinoline is used as the starting material, and intermediate P is obtained by reduction reaction. Intermediate E is oxidized to sulfone by m-CPBA and then undergoes substitution reaction with intermediate P to obtain compound I-iiii of general formula I. Specific implementation method:

[0107] 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.

[0108] Example 1. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0109] 1.1 Preparation of 4-hydroxy-2-(methylthio)pyrimidine-5-carboxylic acid (B)

[0110] At room temperature, ethyl 4-chloro-2-methylthiopyrimidine-5-carboxylate (A) (10.0 g, 43.1 mmol) was dissolved in 100 mL EtOH, 60 mL NaOH aqueous solution (7 mol / L) was added, and the temperature was raised to 90°C for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, 100 mL water was added, and the pH was adjusted to 5 with 6 mol / L hydrochloric acid solution. A white solid precipitated, and the filter cake was washed with a small amount of water and collected. The filter cake was vacuum dried at 40°C for 24 h to obtain 6.4 g of a white solid with a yield of 80%. MS (ESI), m / z (%): 185.1 [MH] - .

[0111] 1.2 Preparation of 4-chloro-2-(methylthio)pyrimidine-5-carbonyl chloride (C)

[0112] At room temperature, 4-hydroxy-2-(methylthio)pyrimidine-5-carboxylic acid (7.0 g, 40 mmol) was dissolved in 70 mL SOCl2, 0.2 mL DMF was added, and the temperature was raised to 80°C for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, toluene was added, and the solution was filtered while hot, the filtrate was collected, the filtrate was concentrated under reduced pressure, and the solution was cooled and dried to obtain a yellow solid, which was directly used in the next step without purification.

[0113] 1.3 Preparation of 4-chloro-N-(2,6-dichlorophenyl)-2-(methylthio)pyrimidine-5-carboxamide (D)

[0114] At room temperature, 2,6-dichloroaniline (7.2 g, 44.7 mmol) was dissolved in 72 mL DCM, pyridine (8 g, 101.3 mmol) was added, and a DCM solution of 4-chloro-2-(methylthio)pyrimidine-5-carbonyl chloride (7.0 g, 31.6 mmol) was slowly dripped into the above reaction solution at -20°C, and reacted at room temperature for 9 hours. After the reaction was completed, 100 mL of ice water was added to the reaction solution, the aqueous layer was extracted with DCM (100 mL×3), the organic layers were combined, and washed with 1 mol / L HCl (200 mL×3) and 1 mol / L NaOH solution (200 mL×3) in sequence, the organic layer was dried with anhydrous sodium sulfate, filtered, the filtrate was evaporated to dryness under reduced pressure, slurried with anhydrous ethanol, filtered, the filter cake was washed with a small amount of ethanol, the filter cake was collected, and dried to obtain 8.5 g of yellow solid, with a yield of 78%. MS (ESI), m / z (%): 347.9 [M+H] + .

[0115] 1.4 Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-(methylthio)pyrimidine-5-carboxamide (E)

[0116] At room temperature, 4-chloro-N-(2,6-dichlorophenyl)-2-(methylthio)pyrimidine-5-carboxamide (3.0 g, 8.65 mmol) was dissolved in 30 mL DMF, and NaH (0.52 g, 21.7 mmol) was slowly added at -20 °C. After stirring for 15 min, anhydrous ethanol (1.19 g, 25.9 mmol) was added to the reaction solution, and the mixture was moved to room temperature for 5 h. After the reaction was completed, the reaction solution was poured into 100 mL of ice water and stirred slowly. A yellow solid precipitated, which was filtered by suction. The filter cake was washed with a small amount of ether, and the filter cake was collected and dried to obtain 1.5 g of off-white solid with a yield of 50%. MS (ESI), m / z (%): 358.1 [M+H] + .

[0117] 1.5 Preparation of 1-methyl-4-(4-nitrophenyl)piperazine (G)

[0118] At room temperature, N-methylpiperazine (11.7 g, 120 mmol), p-fluoronitrobenzene (10.0 g, 70 mmol) and potassium carbonate (20.0 g, 150 mmol) were added to 100 mL of DMF in sequence, and the reaction solution was heated to 100 °C for 2 h. After the reaction was completed, 100 mL of water was added to the reaction solution after cooling, and a yellow solid precipitated. The filter cake was washed with a small amount of water and dried to obtain 16.7 g of a yellow solid with a yield of 91%.

[0119] 1.6 Preparation of 4-(4-methylpiperazin-1-yl)aniline (H)

[0120] At room temperature, in a hydrogen environment, 1-methyl-4-(4-nitrophenyl)piperazine (5.0 g, 20 mmol) was added to 50 mL of ethanol, followed by palladium carbon (wt%) (0.5 g, 5% m / m). After the addition, the reaction solution was heated to 40°C and reacted for 8 h. After the reaction was completed, the filter cake was collected by suction filtration, evaporated to dryness under reduced pressure, and dried to obtain 4.1 g of gray solid with a yield of 94%. MS (ESI), m / z (%): 192.1 [M+H] + .

[0121] 1.7 Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0122] At room temperature, N-(2,6-dichlorophenyl)-4-ethoxy-2-(methylthio)pyrimidine-5-carboxamide (0.2g, 0.56mmol) was dissolved in 10mL toluene, and m-CPBA (0.25g, 1.45mmol) was added. The mixture was reacted at room temperature for 1.5h. DIPEA (0.38g, 2.94mmol) was added to the reaction solution, followed by 4-(4-methylpiperazine-1-yl)aniline (0.11g, 0.59mmol), and the mixture was heated to 80℃ for 10h. After the reaction was completed, 20mL of saturated NaHCO3 aqueous solution was added to the reaction solution, and the mixture was extracted with ethyl acetate (15mL×3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure. The crude product was purified by column chromatography to obtain a yellow solid with a yield of 45%.

[0123] Example 2. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(4-ethylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0124] Using N-methylpiperazine and p-fluoronitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(4-ethylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 48%.

[0125] Example 3. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(piperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0126] Using piperazine and p-fluoronitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(piperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 40%.

[0127] Example 4. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(3-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0128] Using 2-methylpiperazine and p-fluoronitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(3-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 38%.

[0129] Example 5. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-methyl-4-(4-ethylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0130] Using N-ethylpiperazine and 1-fluoro-2-methyl-4-nitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-methyl-4-(4-ethylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 40%.

[0131] Example 6. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-methyl-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0132] Using N-methylpiperazine and 1-fluoro-2-methyl-4-nitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-methyl-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 42%.

[0133] Example 7. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-methyl-4-(piperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0134] Using piperazine and 1-fluoro-2-methyl-4-nitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-methyl-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 43%.

[0135] Example 8. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-methoxy-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0136] Using N-methylpiperazine and 1-fluoro-2-methoxy-4-nitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-methoxy-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 42%.

[0137] Example 9. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-fluoro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0138] Using N-methylpiperazine and 1,2-difluoro-4-nitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-fluoro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 35%.

[0139] Example 10. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[2-fluoro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0140] Using N-methylpiperazine and 2,4-difluoro-1-nitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[2-fluoro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 32%.

[0141] Example 11. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-(trifluoromethyl)-fluoro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0142] Using N-methylpiperazine and 1-fluoro-4-nitro-2-(trifluoromethyl)benzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-(trifluoromethyl)-fluoro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 31%.

[0143] Example 12. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-chloro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0144] Using N-methylpiperazine and 2-chloro-1-fluoro-4-nitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-chloro-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 38%.

[0145] Example 13. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-cyano-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0146] Using N-methylpiperazine and 2-fluoro-5-nitrobenzonitrile as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[3-cyano-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 45%.

[0147] Example 14. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[6-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}pyrimidine-5-carboxamide

[0148] Using N-methylpiperazine and 2-fluoro-5-nitropyridine as raw materials, the intermediate 6-(4-methylpiperazine-1-yl)pyridin-3-amine was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[6-(4-methylpiperazine-1-yl)pyridin-3-yl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 40%.

[0149] Example 15. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(dimethylamino)phenyl]amino}pyrimidine-5-carboxamide

[0150] Using dimethylamine hydrochloride and p-fluoronitrobenzene as raw materials, the intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(dimethylamino)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 45%.

[0151] Example 16. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-[2-(dimethylamino)ethyl]phenyl}amino}pyrimidine-5-carboxamide

[0152] 16.1 Preparation of N,N-dimethyl-2-(4-nitrophenyl)ethan-1-amine

[0153] At room temperature, 1-(2-bromoethyl)-4-nitrobenzene (2.0 g, 8.7 mmol) and anhydrous potassium carbonate (2.4 g, 17.4 mmol) were added to 20 mL of DMF, and then dimethylamine hydrochloride (1.4 g, 17.4 mmol) was added, and the reaction was carried out at room temperature for 6 hours. After the reaction was completed, 100 mL of water was added after the reaction solution was cooled, and a yellow solid was precipitated. The filter cake was washed with a small amount of water and dried to obtain 1.4 g of a yellow solid with a yield of 80%.

[0154] 16.2 Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-[2-(dimethylamino)ethyl]phenyl}amino}pyrimidine-5-carboxamide

[0155] Using N,N-dimethyl-2-(4-nitrophenyl)ethyl-1-amine as a raw material, the intermediate 4-[2-(dimethylamino)ethyl]aniline was prepared according to the synthetic method of 1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-[2-(dimethylamino)ethyl]phenyl}amino}pyrimidine-5-carboxamide was prepared according to the synthetic method of 1.7 in Example 1. The yield was 42%.

[0156] Example 17. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-{[2-(dimethylamino)ethyl](methyl)amino}phenyl}amino}pyrimidine-5-carboxamide

[0157] Using N,N,N'-trimethylethylenediamine and p-fluoronitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-{[2-(dimethylamino)ethyl](methyl)amino}phenyl}amino}pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 40%.

[0158] Example 18. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-[2-(diethylamino)ethoxy]phenyl}amino}pyrimidine-5-carboxamide

[0159] 18.1 Preparation of N,N-diethyl-2-(4-nitrophenoxy)ethan-1-amine

[0160] At room temperature, p-nitrophenol (2.0 g, 14.3 mmol) and Cs2CO3 (7.6 g, 17.2 mmol) were added to 20 mL of dry DMF, and then 2-chloroethyldimethylamine (2.3 g, 8.7 mmol) was slowly added, and the temperature was raised to 100°C for 4 h. After the reaction was completed, the reaction solution was added to 100 mL of ice water, and the aqueous layer was extracted with dichloromethane (3×40 mL). The organic layers were combined, washed with saturated brine (3×40 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness to obtain 2.0 g of gray solid with a yield of 62%. 18.2 Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-[2-(diethylamino)ethoxy]phenyl}amino}pyrimidine-5-carboxamide

[0161] Using N,N-diethyl-2-(4-nitrophenoxy)eth-1-amine as a raw material, the intermediate N was prepared according to the synthetic method of 1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-[2-(diethylamino)ethoxy]phenyl}amino}pyrimidine-5-carboxamide was prepared according to the synthetic method of 1.7 in Example 1. The yield was 41%.

[0162] Example 19. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-[2-(4-methylpiperazin-1-yl)ethoxy]phenyl}amino}pyrimidine-5-carboxamide

[0163] Using 1-(2-chloroethyl)-4-methylpiperazine and p-nitrophenol as raw materials, N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-[2-(4-methylpiperazin-1-yl)ethoxy]phenyl}amino}pyrimidine-5-carboxamide was prepared according to the synthesis method in Examples 18.1-18.2. The yield was 42%.

[0164] Example 20. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino}pyrimidine-5-carboxamide

[0165] Using N-methylpiperazine and 1-bromomethyl-4-nitrobenzene as raw materials, N-(2,6-dichlorophenyl)-4-ethoxy-2-{{4-[2-(4-methylpiperazin-1-yl)ethoxy]phenyl}amino}pyrimidine-5-carboxamide was prepared according to the synthesis method in Examples 16.1-16.2. The yield was 38%.

[0166] Example 21. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-[(4-morpholinylphenyl)amino]pyrimidine-5-carboxamide

[0167] Using morpholine and p-fluoronitrobenzene as raw materials, intermediate H was prepared according to the synthesis method of 1.5-1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-[(4-morpholinylphenyl)amino]pyrimidine-5-carboxamide was prepared according to the synthesis method of 1.7 in Example 1. The yield was 35%.

[0168] Example 22. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(4-acetylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0169] First, N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(piperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide was prepared according to the synthesis method in Example 3. N-(2,6-dichlorophenyl)-4-ethoxy-2-{[4-(piperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide (0.2 g, 0.41 mmol) and N,N-diisopropylethylamine (0.068 g, 0.669 mmol) were added to 5 mL of DCM at 0°C, and acetyl chloride (0.06 g, 0.561 mmol) was slowly added dropwise. After the addition was completed, the mixture was reacted at 25°C for 2 h. After the reaction was completed, 10 mL of water was added to the reaction solution, the aqueous layer was extracted with dichloromethane (10 mL×3), the organic layers were combined, washed with saturated brine (2×20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness and purified by thin layer chromatography to obtain 0.15 g of a white solid with a yield of 60%.

[0170] Example 23. Preparation of ethyl 4-{4-{{5-[(2,6-dichlorophenyl)carbamoyl]-4-ethoxypyrimidin-2-yl}amino}phenyl}piperazine-1-carboxylate

[0171] Using ethyl chloroformate as a raw material, 4-{4-{{5-[(2,6-dichlorophenyl)carbamoyl]-4-ethoxypyrimidin-2-yl}amino}phenyl}piperazine-1-carboxylic acid ethyl ester was prepared according to the synthesis method in Example 22. The yield was 62%.

[0172] Example 24. Preparation of tert-butyl 4-{4-{{5-[(2,6-dichlorophenyl)carbamoyl]-4-ethoxypyrimidin-2-yl}amino}phenyl}piperazine-1-carboxylate

[0173] Using pivaloyl chloride as a raw material, tert-butyl 4-{4-{{5-[(2,6-dichlorophenyl)carbamoyl]-4-ethoxypyrimidin-2-yl}amino}phenyl}piperazine-1-carboxylate was prepared according to the synthesis method in Example 22. The yield was 65%.

[0174] Example 25. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{{[4-(methylsulfonyl)piperazin-1-yl]phenyl}amino}pyrimidine-5-carboxamide

[0175] Using methanesulfonyl chloride as a raw material, N-(2,6-dichlorophenyl)-4-ethoxy-2-{{[4-(methylsulfonyl)piperazin-1-yl]phenyl}amino}pyrimidine-5-carboxamide was prepared according to the synthesis method in Example 22. The yield was 60%.

[0176] Example 26. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl]amino}pyrimidine-5-carboxamide

[0177] 26.1 Preparation of 1-methyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine (O)

[0178] At room temperature, 4-nitro-1H-pyrazole (1.0 g, 9.6 mmol) and 1-methylpiperidin-4-ol (1.0 g, 8.7 mmol) and triphenylphosphine (3.4 g, 13.1 mmol) were added to 20 mL of dry tetrahydrofuran, and DIAD (2.6 g, 13.1 mmol) was added under nitrogen protection and ice bath, and the mixture was reacted at room temperature for 24 h. After the reaction was completed, the reaction was quenched with water (20 mL), the organic solvent was evaporated, and the mixture was extracted with DCM (30 mL × 3), the organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and the solvent was evaporated to obtain a brown-red oily liquid, which was purified by column chromatography to obtain 1.0 g of a white solid with a yield of 65%.

[0179] 26.2 Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-{[1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl]amino}pyrimidine-5-carboxamide

[0180] Using 1-methyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine as a raw material, the intermediate K was prepared according to the synthetic method of 1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-{[1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl]amino}pyrimidine-5-carboxamide was prepared according to the synthetic method of 1.7 in Example 1. The yield was 40%.

[0181] Example 27. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-[(1,2,3,4-tetrahydroisoquinolin-7-yl)amino]pyrimidine-5-carboxamide

[0182] Using 7-nitro-1,2,3,4-tetrahydroisoquinoline as a raw material, the intermediate P was prepared according to the synthetic method of 1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-[(1,2,3,4-tetrahydroisoquinolin-7-yl)amino]pyrimidine-5-carboxamide was prepared according to the synthetic method of 1.7 in Example 1. The yield was 42%.

[0183] Example 28. Preparation of N-(2,6-dichlorophenyl)-4-ethoxy-2-[(2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)amino]pyrimidine-5-carboxamide

[0184] Using 2-methyl-7-nitro-1,2,3,4-tetrahydroisoquinoline as a raw material, the intermediate P was prepared according to the synthetic method of 1.6 in Example 1, and then N-(2,6-dichlorophenyl)-4-ethoxy-2-[(2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)amino]pyrimidine-5-carboxamide was prepared according to the synthetic method of 1.7 in Example 1. The yield was 45%.

[0185] The structures and 1 H-NMR and MS data are shown in Table 1.

[0186] Table 1

[0187]

[0188]

[0189]

[0190]

[0191] The compounds of the present invention were studied for their in vitro enzyme activity, and the results are as follows

[0192] The compounds of the 2-aminopyrimidine-5-carboxamide skeleton of the general formula I of the present invention were tested for in vitro Wee1 enzyme activity using the Lantha screen analysis experiment. The reference substance AZD1775 was prepared by our research group.

[0193] Serial dilution of compounds and preparation of source plates: Dilute compounds to 100 times the highest inhibitor concentration required for the reaction in 100% DMSO. Transfer compounds from test tubes to 96-well storage plates and serially dilute compounds to 5 concentrations. Label this well plate as the source plate. Transfer 40 μL of dilutions from the source plate to a 384-well plate as the intermediate plate. Add 10 μL of kinase solution to each well of the assay plate, except for the control wells without enzyme (10 μL of 1x kinase buffer is added instead). Incubate at room temperature for 10 minutes after shaking. Add 10 μL of substrate solution to each well of the assay plate to start the reaction. Shake. Cover the assay plate and incubate at room temperature for 10 minutes.

[0194] Stop the reaction by adding 20 μL of detection solution to each well of the assay plate. Mix briefly by centrifuge and incubate at room temperature for 60 minutes before reading fluorescence on a plate reader. Copy the RFU values ​​from the Envision program. Calculate the ratio: RFU 520nm / RFU 495nm. Convert the ratio to percent inhibition.

[0195] Inhibition percentage = (max - sample ratio) / (max - min) * 10 ("min" represents the ratio of the no enzyme control, "max" represents the ratio of the DMSO control).

[0196] The data were displayed in MS Excel and the curves were fitted by XLFit excel version 5.4.0.8.

[0197] IC 50 The calculation formula is: Y = Bottom + (Top-Bottom) / (1 + (IC 50 / X)^HillSlope)

[0198] The results of in vitro kinase assays of Wee1 in some examples are shown in Table 2.

[0199] Table 2 IC values ​​of Wee1 kinases in some examples 50 data

[0200] Example <![CDATA[Wee1 IC 50 (nM)]]> Example <![CDATA[Wee1 IC 50 (nM)]]> Example 1 2.0 Example 8 2.71 Example 2 1.98 Example 9 2.22 Example 3 1.71 Example 12 1.99 Example 4 1.89 Embodiment 26 2.11 Example 5 1.04 Embodiment 27 1.55 Example 6 1.42 Embodiment 28 1.47 Example 7 1.35 AZD1775 2.47

[0201] It can be seen from Table 2 that the compounds obtained in the present invention all have corresponding activities, and most of the compounds have outstanding effects. The preliminary results of in vitro inhibition of kinase activity show that the compounds of general formula I to be protected by the present invention have good Wee1 inhibitory activity, and most of the compounds are equivalent to or better than the positive control drug AZD1775.

Claims

1. A 2-aminopyrimidine-5-carboxamide compound, characterized in that: The compound is a 2-aminopyrimidine-5-carboxamide compound of general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof, in, R 1 is unsubstituted or substituted with at least one identical or different R 4 Substituted (C6-C 10 ) aryl, 5-10 membered heteroaryl; R 4 is hydroxy, halogen, nitro, amino, cyano, (C1-C6)alkyl or (C1-C6)alkoxy which is unsubstituted or substituted by at least one of the following substituents which are the same or different, wherein the following substituents are hydroxy, amino or halogen; R 2 is (C1-C6) alkyl or (C3-C7) cycloalkyl, wherein the alkyl includes a straight chain or a branched chain alkyl; R 3 is unsubstituted or substituted with 1 to 3 identical or different R 5 Substituted (C6-C 10 ) aryl, 5-10 membered heteroaryl, tetrahydroquinolyl, tetrahydroisoquinolyl; R 5 is (C1-C6) haloalkyl, hydroxy, halogen, cyano, (C1-C6) alkyl, (C1-C6) alkoxy, -NR 6 R 7 、-(CH2) p NR 6 R 7 、-O(CH2) p NR 6 R 7 、-NH(CH2) p NR 6 R 7 、-NCH3(CH2) p NR 6 R 7 , a 5-10 membered heterocyclic group, the heterocyclic group optionally containing 1-3 double bonds and / or optionally 1-3 identical or different R 8 Substituted, wherein the heteroatoms in the heterocyclic group are 1 to 4 identical or different N, O and / or S; Among them, R 6 and R 7 are the same or different, each independently selected from hydrogen, unsubstituted or substituted with 1-3 identical or different R 9 Substituted (C1-C6) alkyl, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 5-10 membered heterocyclic group; or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 5-10 membered heterocyclic group, the heterocyclic group optionally containing 1-3 double bonds and / or optionally substituted by 1-3 identical or different R 10 Substituted, wherein the heteroatoms in the heterocyclic group are 1 to 4 identical or different N, O and / or S; R 8 , R 9 and R 10 are the same or different and are independently selected from oxo, hydroxyl, halogen, amino, (C1-C6) alkylacyl, (C1-C6) alkoxyformyl, (C1-C6) alkylsulfonyl, (C5-C 10 ) heterocyclic group, free, salified, esterified or amidated carboxyl group, (C1-C6) alkyl group which is unsubstituted or substituted by at least one hydroxyl group, amino group or halogen group, (C1-C6) alkoxy group, (C3-C7) cycloalkyl group; p is an integer from 1 to 4.

2. The 2-aminopyrimidine-5-carboxamide compound according to claim 1, characterized in that: The compound is a 2-aminopyrimidine-5-carboxamide compound of general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: R 1 is unsubstituted or substituted with 1 to 3 identical or different R 4 Substituted phenyl; R 4 is hydroxy, halogen, nitro, amino, cyano, (C1-C4)alkyl or (C1-C4)alkoxy which is unsubstituted or substituted by at least one of the following substituents which are the same or different, wherein the following substituents are hydroxy, amino or halogen; R 2 is (C1-C4) alkyl or (C3-C6) cycloalkyl, wherein the alkyl includes a straight chain or a branched chain alkyl; R 3 is unsubstituted or substituted with 1 to 3 identical or different R 5 Substituted phenyl, 5-6 membered heteroaryl, tetrahydroquinolyl, tetrahydroisoquinolyl; R 5 is trifluoromethyl, hydroxyl, halogen, cyano, (C1-C4) alkyl, (C1-C4) alkoxy, -NR 6 R 7 、-(CH2) p NR 6 R 7 、-O(CH2) p NR 6 R 7 、-NH(CH2) p NR 6 R 7 、-NCH3(CH2) p NR 6 R 7 , a 5-6 membered heterocyclic group, the heterocyclic group optionally containing 1-2 double bonds and / or optionally 1-3 identical or different R 8 Substituted, wherein the heteroatoms in the heterocyclic group are 1 to 3 identical or different N, O and / or S; Among them, R 6 and R 7 are the same or different, each independently selected from hydrogen, unsubstituted or substituted with 1-3 identical or different R 9 Substituted (C1-C4) alkyl, (C3-C6) cycloalkyl, phenyl, 5-6 membered heterocyclic group; or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 5-10 membered heterocyclic group, the heterocyclic group optionally containing 1-2 double bonds and / or optionally substituted by 1-3 identical or different R 10 Substituted, wherein the heteroatoms in the heterocyclic group are 1 to 3 identical or different N, O and / or S; R 8 , R 9 and R 10 are the same or different and are independently selected from oxo, hydroxyl, halogen, amino, (C1-C4) alkylacyl, (C1-C4) alkoxyformyl, (C1-C4) alkylsulfonyl, (C5-C6) heterocyclyl, free, salified, esterified or amidated carboxyl, (C1-C4) alkyl, (C1-C4) alkoxy, (C3-C6) cycloalkyl which is unsubstituted or substituted with at least one hydroxyl, amino or halogen; p is 1, 2 or 3.

3. The 2-aminopyrimidine-5-carboxamide compound according to claim 1, characterized in that: The compound is a 2-aminopyrimidine-5-carboxamide compound of general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: R 1 is unsubstituted or substituted with 1 to 3 identical or different R 4 Substituted phenyl,; R 4 is hydroxy, halogen, nitro, amino, cyano, (C1-C4)alkyl or (C1-C4)alkoxy which is unsubstituted or substituted by at least one of the following substituents which are the same or different, wherein the following substituents are hydroxy, amino or halogen; R 2 is (C1-C4) alkyl or (C3-C4) cycloalkyl, wherein the alkyl includes a straight chain or a branched chain alkyl; R 3 is unsubstituted or substituted with 1 to 3 identical or different R 5 substituted phenyl, pyridyl, pyrazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, tetrahydroquinolyl, tetrahydroisoquinolyl; R 5 is trifluoromethyl, hydroxyl, halogen, cyano, (C1-C3) alkyl, (C1-C3) alkoxy, -NR 6 R 7 、-(CH2) p NR 6 R 7 、-O(CH2) p NR 6 R 7 、-NH(CH2) p NR 6 R 7 、-NCH3(CH2) p NR 6 R 7 , a 5-6 membered heterocyclic group, the heterocyclic group is optionally substituted by 1-2 identical or different R 8 Substitution; wherein the heteroatom in the heterocyclic group contains 1-2 identical or different N, O and / or S; Among them, R 6 and R 7 are the same or different, each independently selected from hydrogen, unsubstituted or substituted with 1-3 identical or different R 9 Substituted (C1-C4) alkyl, (C3-C4) cycloalkyl, phenyl, 5-6 membered heterocyclic group; or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 5-6 membered heterocyclic group, the heterocyclic group optionally containing 1-2 double bonds and / or optionally substituted by 1-3 identical or different R 10 Substituted, wherein the heteroatoms in the heterocyclic group are 1 to 3 identical or different N, O and / or S; R 8 , R 9 and R 10 are the same or different and are independently selected from oxo, hydroxyl, halogen, amino, (C1-C4) alkylacyl, (C1-C4) alkoxyformyl, (C1-C4) alkylsulfonyl, (C5-C6) heterocyclyl, free, salified, esterified or amidated carboxyl, (C1-C4) alkyl, (C1-C4) alkoxy, (C3-C6) cycloalkyl which is unsubstituted or substituted with at least one hydroxyl, amino or halogen; p is 1, 2 or 3.

4. The 2-aminopyrimidine-5-carboxamide compound according to claim 1, characterized in that: The compound is a 2-aminopyrimidine-5-carboxamide compound of general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: R 1 is unsubstituted or substituted with 1 to 3 identical or different R 4 Substituted phenyl; R 4 is hydroxy, halogen, nitro, amino, cyano, (C1-C4)alkyl or (C1-C4)alkoxy which is unsubstituted or substituted by at least one of the following substituents which are the same or different, wherein the following substituents are hydroxy, amino or halogen; R 2 is methyl, ethyl, cyclopropyl, cyclobutyl; R 3 is unsubstituted or substituted with 1 to 3 identical or different R 5 substituted phenyl, pyridyl, pyrazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, tetrahydroquinolyl, tetrahydroisoquinolyl; R 5 For-NR 6 R 7 、-(CH2) p NR 6 R 7 、-O(CH2) p NR 6 R 7 、-NH(CH2) p NR 6 R 7 、-NCH3(CH2) p NR 6 R 7 ; Among them, R 6 and R 7 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl, (C3-C4) cycloalkyl; or R 6 and R 7 Together with the nitrogen atoms to which they are attached, they form p is 1, 2 or 3.

5. The 2-aminopyrimidine-5-carboxamide compound according to claims 1-4, characterized in that: The pharmaceutically acceptable salt of the compound is a salt formed with an acid, wherein 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.

6. A pharmaceutical composition, characterized in that: The composition contains the compound of general formula I according to any one of claims 1 to 4 and its pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof.

7. Use of the compound according to claim 1, characterized in that: Use of the compound according to any one of claims 1 to 4 and its pharmaceutically acceptable salt, hydrate, solvate or prodrug in the preparation of a drug for treating and / or preventing cancer.

8. The use according to claim 7, characterized in that: Use of the compound according to any one of claims 1 to 4 and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof in the preparation of a medicament for preventing or treating a disease associated with Wee1 kinase.

9. The use according to claim 8, characterized in that: The diseases mediated by Wee1 are prevented from being breast cancer, lung cancer, cervical cancer, ovarian cancer, prostate cancer, melanoma and liver cancer.

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