2, 4-diaminopyrimidine compound and application thereof

Abstract

The invention relates to 2, 4-diaminopyrimidine compounds as shown in a general formula I, pharmaceutically acceptable salts, solvates or prodrugs of the 2, 4-diaminopyrimidine compounds, preparation methods of the 2, 4-diaminopyrimidine compounds and the pharmaceutically acceptable salts, solvates or prodrugs of the 2, 4-diaminopyrimidine compounds and the pharmaceutically acceptable salts, solvates or prodrugs of the 2, 4-diaminopyrimidine compounds and the pharmaceutically acceptable salts, solvates or prodrugs of the 2, 4-diaminopyrimidine compounds, and pharmaceutical compositions containing the 2, 4-diaminopyrimidine compounds, the invention further relates to application of the compound shown in the general formula I in preparation of drugs for treating virus infection and malignant tumors. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to a 2,4-diaminopyrimidine compound and its use in preparing a drug for treating and / or preventing viral infection and malignant tumors. Background Art

[0002] Cancer is one of the leading causes of illness and death worldwide. It is estimated that by 2040, the total number of cancer patients worldwide will reach approximately 28 million, necessitating urgent action to combat cancer. Tumor immunotherapy is a novel anti-cancer approach, following surgery, chemotherapy, and targeted therapy. Unlike traditional anti-cancer approaches that directly target tumors, tumor immunotherapy targets tumors by modulating the immune system, restoring or enhancing intrinsic immune capacity. The development of tumor immunotherapies, such as PD-1 / PD-L1 drugs, offers new hope for cancer patients resistant to traditional anti-cancer treatments and represents a milestone in humanity's fight against cancer.

[0003] However, many patients clinically have a low response to tumor immunotherapy approaches, particularly those targeting PD-1 / PD-L1. Furthermore, some patients develop drug resistance and relapse after treatment, presenting challenges for current tumor immunotherapy. With the continued in-depth research on tumor immunotherapy in recent years, mechanisms of tumor immune resistance, tumor heterogeneity, and the complexity of the tumor microenvironment are believed to be the primary drivers of limited treatment efficacy. Studies have revealed that aberrant activation of multiple signaling pathways within tumors leads to overexpression of immunosuppressive molecules, suppressing anti-tumor immune responses. Furthermore, based on the distribution of T cells surrounding the tumor, tumors can be categorized into three basic immune phenotypes: immune-inflammatory, immune-rejective, and immune-desert. Immunoinflammatory tumors are considered "hot tumors," while immune-rejective and immune-desert tumors are both considered "cold tumors." Cold tumors, due to their low levels of T cell recognition and infiltration, are limited by the effectiveness of tumor immunotherapy. Finally, numerous immunosuppressive cells, represented by Terg cells, reside within the tumor microenvironment. These cells secrete immunosuppressive molecules that inhibit T cell activation, ultimately leading to tumor immune escape. Therefore, given the numerous immune escape strategies used by tumors, single PD-1 / PD-L1 drug treatments have limited efficacy. Furthermore, they can also trigger the expression of other immunosuppressive molecules, leading to acquired drug resistance. Therefore, developing new immunomodulatory agents for single or combined therapy is a novel strategy for sustainably restoring and enhancing T cell activity.

[0004] Hematopoietic progenitor kinase 1 (HPK1), also known as mitogen-activated protein kinase kinase kinase kinase 1 (MAP4K1), is a 97-kDa STE20-like serine / threonine kinase. HPK1 was originally cloned in hematopoietic progenitor cells and is highly expressed in a variety of immune cells, including T cells, B cells, and dendritic cells. HPK1's primary physiological function is as a negative regulator of the T cell receptor (TCR). Upon antigen stimulation of the TCR on T cells, HPK1 is recruited to the cell membrane and phosphorylates Ser376 on the SLP76 protein, destabilizing the TCR signaling complex and thus downregulating TCR signaling. Both HPK1 kinase-dead knock-in (HPK1-kd) and HPK1 knockout (HPK1-ko) mouse models have demonstrated that inhibition of HPK1 kinase function enhances T cell function and reduces tumor growth. Importantly, compared to mice with CTLA-4 dysfunction, mice with either HPK1 kinase-dead knock-in or HPK1 knockout did not exhibit fatal inflammation, and due to the restricted expression of HPK1 in hematopoietic cells, the likelihood of systemic side effects was also low. Furthermore, HPK1 inhibitors have the ability to convert "cold tumors" into "hot tumors" through immune activation, and their combination with other immunotherapies, such as PD-1 monoclonal antibodies, can significantly improve the response rate and efficacy of immunotherapy. In recent years, many domestic and foreign companies have developed research pipelines for HPK1 inhibitors, and nine inhibitors have entered clinical trials, but no drugs have yet been marketed. Therefore, the field is in urgent need of new HPK1 inhibitors, especially those with high activity and other excellent properties. Summary of the Invention

[0005] The present invention aims to provide a novel 2,4-diaminopyrimidine derivative and its application.

[0006] To achieve the above purpose, the present invention adopts the following technical solutions:

[0007] A 2,4-diaminopyrimidine compound, wherein the 2,4-diaminopyrimidine compound is a compound represented by general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof,

[0008]

[0009] in,

[0010] R 1 -C(O)NH2, pyrazole which is unsubstituted or substituted with at least one C1-C6 alkyl group;

[0011] Y is CH, N;

[0012] R 2are independently selected from hydrogen, halogen, nitro, hydroxy, amino, cyano, (C 1- C6) alkyl, (C 1- C6) alkoxy, (C3-C 10 )cycloalkoxy;

[0013] L is a single bond, -CH2, -C(O), -S(O), -S(O)2;

[0014] R 3 and R 4 are the same or different, independently selected from hydrogen, (C 1- C6) alkyl, (C 3- C 10 ) cycloalkyl, 4-10 membered heterocyclic group; or R 3 and R 4 Together with the nitrogen atom to which they are attached, they form a 4-10 membered monocyclic heterocyclic group or a bridged bicyclic heterocyclic group;

[0015] R 5 are independently selected from hydrogen, halogen, nitro, hydroxy, amino, cyano, (C 1- C6) alkyl, (C 1- C6) alkoxy;

[0016] m is 0, 1 or 2;

[0017] When m is 0, R 6 is hydrogen; R 7 Selected from (C 1- C6) alkyl, (C3-C 10 )cycloalkyl, (C6-C 10 ) aryl, 4-10 membered heterocyclic group, or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 4-10 membered heterocyclic group;

[0018] When m is 1 or 2, R 6 is selected from hydrogen or methyl; R 7 Selected from hydrogen, (C3-C 10 )cycloalkyl, (C6-C 10 ) aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclyl;

[0019] The R 2 、R 3 、R 4 、R 5 and R 7 (C 1- C6) alkyl, (C 1- C6) alkoxy, (C3-C 10 )cycloalkoxy, heterocyclic, aryl, heteroaryl, R3 and R 4 The heterocyclic group formed, R 6 and R 7 The heterocyclic group is independently optionally substituted by 0-3 identical or different R 8 replace;

[0020] All heterocyclic and heteroaryl groups mentioned above may optionally contain 1-4 identical or different heteroatoms selected from N, O, and S, wherein the heterocyclic group may optionally contain 0-2 carbon-carbon double bonds or carbon-carbon triple bonds;

[0021] R 8 are independently selected from hydrogen, halogen, oxo, nitro, hydroxy, amino, cyano, mercapto, 4-10 membered heterocyclic groups, free, salified, esterified and amidated carboxyl, (C 1- C6) alkylsulfinyl, (C 1- C6) alkylsulfonyl, (C 1- C6) alkylacyl, unsubstituted or substituted with at least one hydroxyl, amino or halogen (C 1- C6) alkyl or (C 1- C6) alkoxy.

[0022] Preferably, the compound of formula I and its pharmaceutically acceptable salt, solvate or prodrug thereof,

[0023] in,

[0024] R 1 -C(O)NH2, pyrazole which is unsubstituted or substituted with at least one C1-C6 alkyl group;

[0025] Y is CH, N;

[0026] R 2 Independently selected from hydrogen, halogen, nitro, hydroxy, amino, cyano, (C1-C4) alkyl, (C1-C4) alkoxy, (C3-C7) cycloalkoxy;

[0027] L is a single bond, -CH2, -C(O), -S(O), -S(O)2;

[0028] R 3 and R 4 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl, (C3-C7) cycloalkyl, 4-6 membered heterocyclyl; or R 3 and R 4 Together with the nitrogen atom to which they are attached, they form a 4-10 membered monocyclic heterocyclic group or a bridged bicyclic heterocyclic group;

[0029] R 5independently selected from hydrogen, halogen, nitro, hydroxy, amino, cyano, (C1-C4) alkyl, (C1-C4) alkoxy;

[0030] m is 0, 1 or 2;

[0031] When m is 0, R 6 is hydrogen; R 7 Selected from (C 1- C4) alkyl, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 4-6 membered heterocyclic group, or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group;

[0032] When m is 1 or 2, R 6 is selected from hydrogen or methyl; R 7 selected from hydrogen, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 5-6 membered heteroaryl, 4-6 membered heterocyclyl;

[0033] The R 2 、R 3 、R 4 、R 5 and R 7 (C 1- C4) alkyl, (C 1- C4)alkoxy, (C3-C7)cycloalkoxy, heterocyclic, aryl, heteroaryl R 3 and R 4 The heterocyclic group formed, R 6 and R 7 The heterocyclic group is independently optionally substituted by 0-3 identical or different R 8 replace;

[0034] All heterocyclic and heteroaryl groups mentioned above may optionally contain 1-4 identical or different heteroatoms selected from N, O, and S, wherein the heterocyclic group may optionally contain 0-2 carbon-carbon double bonds or carbon-carbon triple bonds;

[0035] R 8 Independently selected from hydrogen, halogen, oxo, nitro, hydroxy, amino, cyano, mercapto, 4-6 membered heterocyclic groups, free, salified, esterified and amidated carboxyl, (C1-C4) alkylsulfinyl, (C1-C4) alkylsulfonyl, (C1-C4) alkylacyl, (C1-C4) alkyl or (C1-C4) alkoxy groups which are unsubstituted or substituted by at least one hydroxy, amino or halogen group.

[0036] Further preferably, the compound of formula I and its pharmaceutically acceptable salt, solvate or prodrug thereof,

[0037] in,

[0038] R 1 is -C(O)NH2, 1H-pyrazol-4-yl, 1-methyl-1H-pyrazol-4-yl;

[0039] Y is CH, N;

[0040] R 2 independently selected from hydrogen, halogen, (C1-C4)alkyl, (C1-C4)alkoxy, (C3-C7)cycloalkoxy;

[0041] L is a single bond, -CH2, or -C(O);

[0042] R 3 and R 4 are the same or different and are independently selected from hydrogen, a 4-6 membered heterocyclic group; or R 3 and R 4 Together with the nitrogen atom to which they are attached, they form a 4-10 membered monocyclic heterocyclic group or a bridged bicyclic heterocyclic group;

[0043] R 5 independently selected from hydrogen, halogen, (C1-C4)alkyl, (C1-C4)alkoxy;

[0044] m is 0, 1 or 2;

[0045] When m is 0, R 6 is hydrogen; R 7 Selected from (C 1- C4) alkyl, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 4-6 membered heterocyclic group, or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group;

[0046] When m is 1 or 2, R 6 is selected from hydrogen or methyl; R 7 selected from hydrogen, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 5-6 membered heteroaryl, 4-6 membered heterocyclyl;

[0047] The R 2 、R 3 、R 4 、R 5 and R 7 (C 1- C4) alkyl, (C 1- C4)alkoxy, (C3-C7)cycloalkoxy, heterocyclyl, aryl, heteroaryl, R 3 and R 4The heterocyclic group formed, R 6 and R 7 The heterocyclic group is independently optionally substituted by 0-3 identical or different R 8 replace;

[0048] All heterocyclic and heteroaryl groups mentioned above may optionally contain 1-4 identical or different heteroatoms selected from N, O, and S, wherein the heterocyclic group may optionally contain 0-2 carbon-carbon double bonds or carbon-carbon triple bonds;

[0049] R 8 Independently selected from hydrogen, halogen, oxo, hydroxy, cyano, (C1-C4) alkyl, (C1-C4) alkoxy, 4-6 membered heterocyclyl, (C1-C4) alkyl or (C1-C4) alkoxy substituted by at least one halogen.

[0050] Still further preferably, the compound of formula I and its pharmaceutically acceptable salt, solvate or prodrug thereof,

[0051] in,

[0052] R 1 is -C(O)NH2;

[0053] Y is CH;

[0054] R 2 independently selected from hydrogen, halogen, methyl, methoxy, ethoxy;

[0055] L is a single bond, -CH2, or -C(O);

[0056] R 3 and R 4 Together with the nitrogen atom to which they are attached, they form a 4-10 membered monocyclic heterocyclic group or a bridged bicyclic heterocyclic group;

[0057] R 5 independently selected from hydrogen, halogen, methyl, methoxy, ethoxy;

[0058] m is 0, 1 or 2;

[0059] When m is 0, R 6 is hydrogen; R 7 Selected from (C 1- C4) alkyl, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 4-6 membered heterocyclic group, or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group;

[0060] When m is 1 or 2, R 6 is selected from hydrogen; R 7selected from hydrogen, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 5-6 membered heteroaryl, 4-6 membered heterocyclyl;

[0061] The R 3 、R 4 and R 7 Heterocyclic group, aryl group, heteroaryl group, R 3 and R 4 The heterocyclic group is independently optionally substituted by 0-3 identical or different R 8 replace;

[0062] All heterocyclic and heteroaryl groups mentioned above may optionally contain 1-4 identical or different heteroatoms selected from N, O, and S, wherein the heterocyclic group may optionally contain 0-2 carbon-carbon double bonds or carbon-carbon triple bonds;

[0063] R 8 Independently selected from hydrogen, halogen, oxo, hydroxy, cyano, methyl, methoxy, trifluoromethyl.

[0064] More preferably, the compound of formula I and its pharmaceutically acceptable salt, solvate or prodrug thereof,

[0065] in,

[0066] R 1 is -C(O)NH2;

[0067] Y is CH;

[0068] R 2 independently selected from hydrogen, halogen, methyl, methoxy, ethoxy;

[0069] L is a single bond, -CH2, or -C(O);

[0070] R 3 and R 4 Together with the nitrogen atoms to which they are attached, they form:

[0071] R 5 independently selected from hydrogen, halogen, methyl, methoxy, ethoxy;

[0072] m is 0 or 1;

[0073] When m is 0, R 6 is hydrogen; R 7 Selected from (C 1- C4) alkyl, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 4-6 membered heterocyclic group, or R 6 and R 7Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group;

[0074] When m is 1, R 6 is selected from hydrogen; R 7 Selected from (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 5-6 membered heteroaryl, 4-6 membered heterocyclyl;

[0075] The R 7 The heterocyclic group, aryl group and heteroaryl group in the 8 replace;

[0076] All heterocyclic and heteroaryl groups mentioned above may optionally contain 1-4 identical or different heteroatoms selected from N, O, and S, wherein the heterocyclic group may optionally contain 0-2 carbon-carbon double bonds or carbon-carbon triple bonds;

[0077] R 8 Independently selected from hydrogen, halogen, oxo, hydroxy, cyano, methyl, methoxy, trifluoromethyl.

[0078] More preferably, the 2-aminopyrimidine compound of general formula I and its pharmaceutically acceptable salt, solvate or prodrug thereof:

[0079] 2-({4-[(4-acetylphenyl)carbamoyl]phenyl}amino)-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0080] 2-({4-[N-methyl-(carbamoyl)phenyl]amino}-4-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidine-5-carboxamide

[0081] 2-{[4-(Morpholine-4-carbonyl)phenyl]amino}-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0082] 2-({4-[(4-oxocyclohexyl)carbamoyl]phenyl}amino)-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0083] 2-{[4-(Cyclopropylcarbamoyl)phenyl]amino}-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0084] 2-({[(4-morpholinylamino)carbonyl]phenyl}amino)-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0085] 2-[(4-{[2-(4-morpholinyl)ethyl]carbamoyl}phenyl)amino]-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0086] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0087] 2-({4-[(2-phenylethyl)carbamoyl]phenyl}amino)-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0088] 2-{[4-(Benzylcarbamoyl)-3-methylphenyl]-amino}-4-{[-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0089] 2-({4-[(pyridin-2-ylmethyl)carbamoyl]phenyl}amino)-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0090] 2-{[4-(Benzylcarbamoyl)-3-fluorophenyl]-amino}-4-{[-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0091] 2-({4-[(4-fluorobenzyl)carbamoyl]phenyl}amino)-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0092] 2-({4-[(4-chlorobenzyl)carbamoyl]phenyl}amino)-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0093] 2-({4-[(4-methoxybenzyl)carbamoyl]phenyl}amino)-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0094] 2-({4-[(3-fluorobenzyl)carbamoyl]phenyl}amino)-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0095] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[2-methoxy-4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0096] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[2-methoxy-4-(3,4,5-trimethylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0097] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[4-(3,4,5-trimethylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0098] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[6-(4-methylpiperazin-1-yl)pyridin-3-yl]amino}pyrimidine-5-carboxamide

[0099] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[2-methoxy-4-(4-methylpiperazine-1-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0100] 2-{[4-(cyclohexylcarbamoyl)phenyl]amino}-4-{[2-methoxy-4-(4-methylpiperazine-1-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0101] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[2-methoxy-4-(8-methyl-3,8-diazabicyclo[3.2.1]octane-3-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0102] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[4-(3,4-dimethylpiperazine-1-carbonyl)-2-methoxyphenyl]amino}pyrimidine-5-carboxamide

[0103] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[2-methoxy-4-(morpholine-4-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0104] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[4-(4-ethylpiperazine-1-carbonyl)-2-methoxyphenyl]amino}pyrimidine-5-carboxamide

[0105] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[2-methoxy-4-(4-methyl-1,4-diaza-1-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0106] 2-({4-[(4-fluorobenzyl)carbamoyl]phenyl}amino)-4-{[2-methoxy-4-(8-methyl-3,8-diazabicyclo[3.2.1]octane-3-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0107] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[2-ethoxy-4-(8-methyl-3,8-diazabicyclo[3.2.1]octane-3-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0108] 2-{[4-(cyclohexylcarbamoyl)phenyl]amino}-4-{[2-ethoxy-4-(4-methylpiperazine-1-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0109] 2-({4-[(4-fluorobenzyl)carbamoyl]phenyl}amino)-4-{[2-ethoxy-4-(morpholine-4-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0110] 4-({2-ethoxy-4-[4-(oxetan-3-yl)piperazine-1-carbonyl]phenyl}amino)-2-({4-[(4-fluorobenzyl)carbamoyl]phenyl}amino)pyrimidine-5-carboxamide

[0111] 2-({4-[(cyclopropylmethyl)carbamoyl]phenyl}amino)-4-{[2-methoxy-4-(8-methyl-3,8-diazabicyclo[3.2.1]octane-3-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0112] 2-({4-[(cyclohexylmethyl)carbamoyl]phenyl}amino)-4-{[2-methoxy-4-(8-methyl-3,8-diazabicyclo[3.2.1]octane-3-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0113] 2-({4-[(cyclopentylmethyl)carbamoyl]phenyl}amino)-4-{[2-methoxy-4-(8-methyl-3,8-diazabicyclo[3.2.1]octane-3-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0114] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[2-methoxy-4-(6-methyl-3,6-diazabicyclo[3.1.1]heptane-3-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0115] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[4-(8-oxa-3-azabicyclo[3.2.1]octane-3-carbonyl)-2-methoxyphenyl]amino}pyrimidine-5-carboxamide

[0116] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-{[2-methoxy-4-(3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0117] 2-{[4-(benzylcarbamoyl)phenyl)amino)-4-((2-methoxy-4-((1S,4S)-5-methyl-2,5-diazabicyclo[2.2.1]heptane-2-carbonyl)phenyl)amino)pyrimidine-5-carboxamide

[0118] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-({2-methoxy-4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)pyrimidine-5-carboxamide

[0119] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-({2-methoxy-4-[(8-methyl-3,8-diazabicyclo[3.2.1]octan-3-yl)methyl]phenyl}amino)pyrimidine-5-carboxamide

[0120] 2-{[4-(Benzylcarbamoyl)phenyl]amino}-4-({2-methoxy-4-[(3,4,5-trimethylpiperazin-1-yl)methyl]phenyl}amino)pyrimidine-5-carboxamide

[0121] N-Benzyl-4-{[4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}-5-(1H-pyrazol-4-yl)pyrimidin-2-yl]amino}benzamide

[0122] N-Benzyl-4-{[4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}-5-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-yl]amino}benzamide

[0123] N-benzyl-4-{[4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}-5-(1H-pyrazol-4-yl)pyrimidin-2-yl]amino}benzamide.

[0124] Unless otherwise indicated, the term "halogen" used in the present invention refers to fluorine, chlorine or bromine; "alkyl" refers to a straight-chain or branched alkyl; "cycloalkyl" refers to a substituted or unsubstituted cycloalkyl; "alkoxy" refers to a straight-chain or branched alkoxy; "aryl" refers to a phenyl group with no substituent or with a substituent; "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, such as pyridyl, pyrazolyl, imidazolyl, furyl, thienyl, Pyrrolyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl, etc.; "heterocyclyl" refers to a monocyclic or bridged ring system containing one or more heteroatoms selected from N, O, and S, such as pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, 3,8-diazabicyclo[3.2.1]octanyl, 3,6-diazabicyclo[3.1.1]heptyl, 8-oxa-3-azabicyclo[3.2.1]octanyl, (1S,4S)-2,5-diazabicyclo[2.2.1]heptyl.

[0125] According to some common methods in the field of the present invention, the compound of the general formula I of the present invention can be reacted with an acid to form a pharmaceutically acceptable salt thereof. 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 or aspartic acid.

[0126] The present invention also includes prodrugs of the derivatives of the present invention. Prodrugs of the derivatives of the present invention are derivatives of Formula I that may themselves have weak or even no activity, but after administration, are converted to the corresponding biologically active form under physiological conditions (e.g., by metabolism, solvolysis, or other means).

[0127] The present invention also includes pharmaceutical compositions comprising compounds of formula I and their pharmaceutically acceptable salts and / or solvates as active ingredients and pharmaceutically acceptable carriers; 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.

[0128] The carriers used in the pharmaceutical compositions of the present invention are common types available in the pharmaceutical field, including: binders, lubricants, disintegrants, solubilizers, diluents, stabilizers, suspending agents, pigments, flavoring agents, etc. for oral preparations; pH adjusters, osmotic pressure regulators, solubilizers, stabilizers, etc. for injectable preparations; and bases, diluents, lubricants, preservatives, etc. for topical preparations. Pharmaceutical preparations can be administered orally, parenterally (e.g., intravenously, subcutaneously, intraperitoneally, etc.), or topically (e.g., ophthalmically, nasally, sublingually, dermally, etc.). If certain drugs are unstable under gastric conditions, they can be formulated into enteric-coated tablets.

[0129] The exact amount of the compound of the present invention required to treat viral infection or malignancy 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. An appropriate effective amount can be determined by one of ordinary skill in the art using only routine experimentation.

[0130] The invention is used to treat and / or prevent HPK1-mediated diseases including but not limited to viral infection, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, gastric cancer, skin cancer, ovarian cancer, cervical cancer, kidney cancer, head and neck cancer, lymphoma, leukemia, and melanoma.

[0131] The compound can be administered in an amount ranging from about 0.1 to 160 mg / kg body weight per day, preferably 1 to 60 mg / kg body weight per day. It will be appreciated that the dosage may vary depending on the patient's needs, the severity of the viral infection or malignancy being treated, and the specific compound being used. Furthermore, it will be appreciated 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 circumstances. If desired, the daily dose may also be administered in multiple doses, for example, 2-4 times per day.

[0132] Mammal means a human or an animal.

[0133] The amount of active ingredient, i.e., the compound according to the present invention, in the pharmaceutical composition and its unit dosage form can vary depending on the specific application, the potency of the specific compound and the desired concentration. Generally speaking, the active ingredient content will be between 0.5% and 90% by weight of the total weight of the composition.

[0134] In combination therapy, the compound of the present invention and the other compound may be administered simultaneously or intermittently. When administered simultaneously, the compound of the present invention and the other compound may be combined in a single pharmaceutical composition or in separate compositions.

[0135] The following examples and preparations are provided to 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.

[0136] The following synthetic routes describe the preparation of derivatives of formula I of the present invention, all of which are prepared by the methods described in these synthetic routes, by methods well known to those skilled in the art of organic chemistry, or are commercially available. All final compounds of the present invention are prepared by the methods described in these synthetic routes or by methods similar thereto, which are well known to those skilled in the art of organic chemistry. All variable factors used in these synthetic routes are as defined below or as defined in the claims.

[0137] According to the compound of formula I of the present invention, in routes 1 to 4, taking the following compound as an example, the substituent R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , Y, and L have the same definitions as in the claims.

[0138] When R 1 is -C(O)NH2, L is a single bond and is located in the para position of the amino group. Some compounds in the general formula can be synthesized according to Route 1:

[0139]

[0140] In route 1, substituted 4-fluoronitrobenzene or 2-fluoro-5-nitropyridine 1 is used as a raw material to obtain intermediate 3 through substitution and reduction reaction; uracil-5-carboxylic acid 4 is used as a raw material to obtain intermediate 6 through chlorination and condensation reaction; intermediate 6 and intermediate 3 undergo aromatic nucleophilic substitution reaction to obtain intermediate 7; substituted 4-nitrobenzoic acid is used as a raw material to obtain intermediate 10 through condensation and reduction reaction; intermediate 7 and intermediate 10 undergo aromatic nucleophilic substitution reaction to obtain part of the target compound.

[0141] When R 1 is -C(O)NH2, L is -C(O), and is located in the para position of the amino group. Some compounds in the general formula can be synthesized according to Route 2:

[0142]

[0143] In route 2, substituted 4-nitrobenzoic acid is used as the starting material, and intermediate 13 is obtained through condensation and reduction reaction; intermediate 13 and intermediate 6 obtained in route 1 are subjected to aromatic nucleophilic substitution reaction to obtain intermediate 14; intermediate 14 and intermediate 10 obtained in route 1 are further subjected to aromatic nucleophilic substitution reaction to obtain part of the target compound II.

[0144] When R 1 Where -C(O)NH2, L is CH2 and is located in the para position of the amino group, some compounds of the general formula can be synthesized according to Route 3:

[0145]

[0146] In route 3, intermediate 19 is prepared from substituted 4-nitrobenzoic acid through reduction, bromination, nucleophilic substitution and reduction reactions; intermediate 19 and intermediate 6 obtained in route 1 undergo aromatic nucleophilic substitution reaction to prepare intermediate 20; intermediate 20 and intermediate 10 obtained in route 1 undergo further aromatic nucleophilic substitution reaction to prepare part of the target compound (III).

[0147] When R 1 is 1H-pyrazol-4-yl or 1-methyl-1H-pyrazol-4-yl, L is located in the para position of the amino group, and some compounds in the general formula can be synthesized according to route 4:

[0148]

[0149] In route 4, 2,4-dichloro-5-bromo-pyrimidine 21 is used as the raw material, and undergoes an aromatic nucleophilic substitution reaction with the intermediate 3 obtained in route 1 and the intermediate 13 obtained in route 2 to obtain intermediate 22; intermediate 22 and the intermediate 10 obtained in route 1 undergo an aromatic nucleophilic substitution reaction to obtain intermediate 23; intermediate 23 undergoes a coupling reaction to obtain part of the target compound (IV). DETAILED DESCRIPTION

[0150] The following examples describe methods for preparing some of the compounds described. It should be understood that the following methods, as well as other methods known to those skilled in the art, are applicable to the preparation of all compounds described herein. The examples are intended to illustrate, not to limit, the scope of the invention. Proton NMR spectra of the compounds were measured using a Bruker ARX-600, and mass spectra were measured using a Waters ACQUITY triple quadrupole liquid chromatography-mass spectrometer. All reagents used were of analytical or chemical grade.

[0151] Example 1. Preparation of 2-({4-[(4-acetylphenyl)carbamoyl]phenyl}amino)-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidine-5-carboxamide

[0152] 1.1 Synthesis of 1-methyl-4-(4-nitrophenyl)piperazine (2)

[0153] In an ice-water bath at 0°C, add p-fluoronitrobenzene (5.0 g, 35.4 mmol), N-methylpiperazine (3.9 g, 39.0 mmol), and potassium carbonate (5.9 g, 42.5 mmol) to N,N-dimethylformamide (50 mL). The mixture was allowed to react at 0°C for 2 h. After completion of the reaction, the reaction solution was slowly poured into water (250 mL) and stirred at room temperature for 0.5 h. The mixture was filtered, the filter cake was washed with water, and dried to obtain 6.8 g of a brown solid (86.7% yield).

[0154] 1.2 Synthesis of 4-(4-methylpiperazin-1-yl)aniline (3)

[0155] 1-Methyl-4-(4-nitrophenyl)piperazine (5.0 g, 22.6 mmol) and 55% palladium on carbon (0.5 g) were added to methanol (50 mL) at room temperature under a hydrogen atmosphere and allowed to react for 6 h. After completion of the reaction, the mixture was filtered through a pad of celite and the filter cake was washed with methanol (50 mL). The filtrate was collected, the organic solvent was evaporated under reduced pressure, and dried to yield 4.2 g of a dark brown solid (97.7% yield).

[0156] 1.3 Synthesis of 2,4-dichloropyrimidine-5-carboxamide (6)

[0157] At room temperature, the starting material, 2,4-dichloropyrimidine-5-carboxylic acid (10.0 g, 64 mmol), was added to phosphorus oxychloride (100 mL). Phosphorus pentachloride (44.0 g, 211 mmol) was slowly added. After addition, the mixture was heated to reflux and reacted for 10 hours. After the reaction was complete, the phosphorus oxychloride was evaporated under reduced pressure. Dry dichloromethane (100 mL) was added to the residue. Ammonia water (10 mL, 25% wt) was slowly added dropwise at -20°C. After completion, the mixture was maintained at -20°C for 1 hour. After the reaction was complete, the mixture was filtered, the filter cake was washed with a small amount of water, and dried to yield 10.5 g of an off-white solid (85.4% yield).

[0158] 1.4 Synthesis of 2-chloro-4-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidine-5-carboxamide (7)

[0159] 2,4-Dichloropyrimidine-5-carboxamide (4.0 g, 20.9 mmol), 4-(4-methylpiperazin-1-yl)aniline (4.0 g, 20.9 mmol), and potassium carbonate (3.5 g, 25.0 mmol) were added to N,N-dimethylformamide (40 mL) in an ice-water bath at 0°C. The mixture was allowed to react at 0°C for 3 h. After completion of the reaction, the reaction solution was slowly poured into water (200 mL) and stirred at room temperature for 0.5 h. The mixture was filtered, the filter cake was washed with water, and dried to obtain 6.0 g of a yellow solid with a yield of 69.0%.

[0160] 1.5 Synthesis of N-(4-acetylphenyl)-4-nitrobenzamide (9)

[0161] At room temperature, p-nitrobenzoic acid (2.6 g, 15.5 mmol), EDCI (4.4 g, 22.2 mmol), HOBT (3.0 g, 22.2 mmol), and triethylamine (6.0 g, 59.2 mmol) were added to N,N-dimethylformamide (25 mL). The mixture was stirred at room temperature for 0.5 h, followed by the addition of p-aminoacetophenone (2.0 g, 14.8 mmol). The reaction was allowed to proceed at room temperature for 2 h. After completion of the reaction, the reaction solution was slowly poured into water (125 mL), stirred at room temperature for 0.5 h, filtered, and the filter cake was washed with water and dried to obtain 2.8 g of a yellow solid (yield 66.7%).

[0162] 1.6 Synthesis of N-(4-acetylphenyl)-4-aminobenzamide (10)

[0163] N-(4-Acetylphenyl)-4-nitrobenzamide (2.0 g, 7.0 mmol) and 55% palladium on carbon (0.2 g) were added to methanol (20 mL) at room temperature under a hydrogen atmosphere and allowed to react for 6 h. After completion of the reaction, the mixture was filtered through a pad of celite and the filter cake was washed with methanol (20 mL). The filtrate was collected, the organic solvent was evaporated under reduced pressure, and dried to obtain 1.7 g of a dark brown solid (95.0% yield).

[0164] 1.7 Synthesis of 4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}-2-({4-[(4-acetylphenyl)carbamoyl]phenyl}amino)-pyrimidine-5-carboxamide (I-1)

[0165] At room temperature, N-(4-acetylphenyl)-4-aminobenzamide (0.11 g, 0.4 mmol) and 2-chloro-4-((4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidine-5-carboxamide (0.15 g, 0.4 mmol) were added to isopropanol (2 mL), and trifluoroacetic acid (0.1 g, 0.9 mmol) was slowly added dropwise. After the addition was complete, the temperature was raised to 85°C and the reaction was allowed to react for 10 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and water (2 mL) was added to the residue. The pH was adjusted to 9-10 with 4 M aqueous sodium hydroxide solution, filtered, the filter cake was washed with water, and dried to obtain a crude product. The crude product was purified by preparative thin-layer chromatography to obtain 0.05 g of a light yellow solid, with a yield of 20.5%.

[0166] According to the synthesis method of Example 1, the compounds of Examples 2 to 20 were synthesized using substituted 4-nitrobenzoic acid methyl ester as raw material. The structures of some of the compounds in the examples are as follows: 1 H-NMR data are shown in Table 1.

[0167] Table 1

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174] Example 21. Preparation of 2-{[4-(benzylcarbamoyl)phenyl]amino}-4-{[4-(4-methylpiperazine-1-carbonyl)phenyl]amino}pyrimidine-5-carboxamide

[0175] 2.1 Synthesis of (4-methylpiperazin-1-yl)(4-nitrophenyl)methanone (12)

[0176] At room temperature, p-nitrobenzoic acid (1.0 g, 6.0 mmol) and N,N-dimethylformamide (0.44 g, 6.0 mmol) were added to dry dichloromethane (5 mL). Sulfonyl chloride (1.4 g, 12.0 mmol) was slowly added dropwise. After addition, the mixture was heated to 40°C and allowed to react for 4 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove the residual thionyl chloride. Dry dichloromethane (5 mL) was added to the residue and diluted for later use. N-methylpiperazine (0.6 g, 6.0 mmol) and triethylamine (2.4 g, 24.0 mmol) were added to dry dichloromethane at room temperature. The prepared dilute solution of the acid chloride in dichloromethane was added dropwise to the reaction mixture at -20°C. After addition, the mixture was transferred to room temperature and allowed to react for 1 h. After the reaction was completed, water (10 mL) was slowly added to the reaction solution to quench the reaction. The organic layer was collected and separated, washed with saturated brine (10 mL×2), dried over anhydrous sodium sulfate, filtered, and the organic liquid was collected and concentrated under reduced pressure to obtain 1.3 g of a yellow solid with a yield of 86.7%.

[0177] 2. Synthesis of 2-(4-aminophenyl)(4-methylpiperazin-1-yl)methanone (13)

[0178] At room temperature, (4-methylpiperazin-1-yl)(4-nitrophenyl)methanone (1.3 g, 5.2 mmol), ferric chloride hexahydrate (0.21 g, 0.8 mmol), and activated carbon (0.02 g, 1.6 mmol) were added to 1,4-dioxane (13 mL). The mixture was stirred at 70°C for 0.5 h. 80% hydrazine hydrate (4.9 g, 78 mmol) was slowly added dropwise to the reaction mixture. After addition, the mixture was heated to 80°C and allowed to react for 5 h. After completion of the reaction, the mixture was filtered while hot through a pad of celite and concentrated under reduced pressure. Water (25 mL) was added to the residue, and the mixture was extracted with dichloromethane (10 mL x 3). The organic layer was collected, washed with saturated brine (15 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield 0.9 g of an orange oil (78.9% yield).

[0179] 2.3 Synthesis of 2-chloro-4-{[4-(4-methylpiperazine-1-carbonyl)phenyl]amino}pyrimidine-5-carboxamide (14)

[0180] (4-Aminophenyl)(4-methylpiperazin-1-yl)methanone (0.9 g, 2.4 mmol), 2,4-dichloropyrimidine-5-carboxamide (0.46 g, 2.4 mmol), and triethylamine (0.27 g, 2.6 mmol) were added to ethanol (9 mL) at room temperature and the mixture was heated to 45°C for 2 h. After completion of the reaction, the mixture was filtered while hot, washed with a small amount of ethanol, and oven-dried to yield 1.1 g of a yellow solid (71.4% yield).

[0181] 2.4 Synthesis of 2-{[4-(benzylcarbamoyl)phenyl]amino}-4-{[4-(4-methylpiperazine-1-carbonyl)phenyl]amino}pyrimidine-5-carboxamide (II-2)

[0182] 2-Chloro-4-{[4-(4-methylpiperazine-1-carbonyl)phenyl]amino}pyrimidine-5-carboxamide (0.10 g, 0.27 mmol) and 4-amino-N-benzylbenzamide (0.06 g, 0.27 mmol) were added to isopropanol (2 mL) at room temperature, and trifluoroacetic acid (0.06 g, 0.53 mmol) was slowly added dropwise. After the addition was complete, the temperature was raised to 85°C and the reaction was allowed to react for 8 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and water (2 mL) was added to the residue. The pH was adjusted to 9-10 with 4M aqueous sodium hydroxide solution, filtered, the filter cake was washed with water, and dried to obtain the crude product. The crude product was purified by preparative thin-layer chromatography to obtain 0.04 g of a light yellow solid, with a yield of 26.7%.

[0183] According to the synthesis method of Example 21, the compounds of Examples 22 to 39 were synthesized using substituted 4-nitrobenzoic acid methyl ester as raw material. The structures of some of the compounds in the examples, MS, 1The H-NMR data are shown in Table 2.

[0184] Table 2

[0185]

[0186]

[0187]

[0188]

[0189]

[0190]

[0191] Example 40. Synthesis of 2-{[4-(benzylcarbamoyl)phenyl]amino}-4-({2-methoxy-4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)pyrimidine-5-carboxamide

[0192] 3.1 Synthesis of (3-methoxy-4-nitrophenyl)methanol (16)

[0193] 3-Methoxy-4-nitrobenzoic acid (5.0 g, 25.4 mmol) was added to anhydrous tetrahydrofuran (25 mL) at -20°C. A 1.0 mol / L borane solution in tetrahydrofuran (38.6 mL, 38.0 mmol) was slowly injected under a nitrogen atmosphere. The mixture was allowed to react at room temperature for 5 h. After completion, methanol (50 mL) was slowly added dropwise to the reaction mixture under an ice-water bath to quench the reaction. The reaction mixture was concentrated under reduced pressure to obtain 4.5 g of a light yellow oil with a yield of 97.0%.

[0194] 3.2 Synthesis of 4-(bromomethyl)-2-methoxy-1-nitrobenzene (17)

[0195] Under an ice-water bath, 3-methoxy-4-nitrophenyl)methanol (4.5 g, 24.6 mmol) was added to dichloromethane (45 mL). Phosphorus tribromide (8.0 g, 29.5 mmol) was slowly added dropwise to the reaction solution. After addition, the mixture was transferred to room temperature and allowed to react for 1 h. After completion of the reaction, water (45 mL) was slowly added to the reaction solution under an ice-water bath to quench the reaction. The mixture was extracted with dichloromethane (15 mL x 3). The organic solution was collected, washed with saturated brine (45 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 5.1 g of an orange oil with a yield of 84.3%.

[0196] 3.3 Synthesis of 1-(3-methoxy-4-nitrobenzyl)-4-methylpiperazine (18)

[0197] At room temperature, 4-(Bromomethyl)-2-methoxy-1-nitrobenzene (1.0 g, 4.1 mmol) and triethylamine (0.49 g, 4.9 mmol) were added to dichloromethane (10 mL). N-methylpiperazine (0.45 g, 4.5 mmol) was added dropwise to the reaction solution. The temperature was raised to 35°C and the reaction was allowed to react for 1 h. After the reaction was complete, water (15 mL) was slowly added to the reaction solution. The mixture was extracted with dichloromethane (5 mL x 3). The organic layer was collected, washed with saturated brine (15 mL x 2), dried over anhydrous sodium sulfate, filtered, and the collected organic solution was concentrated under reduced pressure to obtain 0.79 g of an orange solid with a yield of 78.7%.

[0198] 3.4 Synthesis of 2-methoxy-4-((4-methylpiperazin-1-yl)methyl)aniline (19)

[0199] 1-(3-Methoxy-4-nitrobenzyl)-4-methylpiperazine (0.79 g, 2.9 mmol) and 55% palladium on carbon (0.079 g) were added to methanol (16 mL) at room temperature and heated to 45°C under a hydrogen atmosphere for 3 h. After completion, the reaction was filtered through celite and the filter cake rinsed with a small amount of methanol. The organic solution was collected and concentrated under reduced pressure to afford 0.68 g of a brown oil (97.1% yield).

[0200] 3.5 Synthesis of 2-methoxy-4-((4-methylpiperazin-1-yl)methyl)aniline (20)

[0201] 2-Methoxy-4-((4-methylpiperazin-1-yl)methyl)aniline (0.68 g, 2.9 mmol) and 2,4-dichloropyrimidine-5-carboxamide (0.55 g, 2.9 mmol) were added to ethanol (7 mL) at room temperature and heated to 45°C for 5 h. After completion of the reaction, the mixture was filtered while hot, rinsed with a small amount of ethanol, and oven-dried to yield 0.77 g of a dark yellow solid (68.1% yield).

[0202] 3.6 Synthesis of N-methyl-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}-2-{[4-(benzylcarbamoyl)phenyl]amino}pyrimidine-5-carboxamide (III-1)

[0203] At room temperature, 2-methoxy-4-((4-methylpiperazin-1-yl)methyl)aniline (0.1 g, 0.26 mmol) and 4-amino-N-benzylbenzamide (0.06 g, 0.26 mmol) were added to isopropanol (2 mL). Trifluoroacetic acid (0.06 g, 0.51 mmol) was slowly added dropwise. After the addition was complete, the temperature was raised to 85°C and the reaction was allowed to react for 8 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and water (2 mL) was added to the residue. The pH was adjusted to 9-10 with 4 M aqueous sodium hydroxide solution. The product was filtered, the filter cake was washed with water, and dried to obtain the crude product. The crude product was purified by preparative thin-layer chromatography to obtain 0.05 g of a light yellow solid in a yield of 36.4%.

[0204] According to the synthesis method of Example 40, the compounds of Example 41 and Example 42 were synthesized using substituted 3-methoxy-4-nitrobenzoic acid as raw materials. The structures of the compounds of the examples are: 1 H-NMR data are shown in Table 3.

[0205] Table 3

[0206]

[0207] Example 43 Synthesis of N-benzyl-4-[(4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}-5-(1H-pyrazol-4-yl)pyrimidin-2-yl)amino]benzamide

[0208] 4.1 Synthesis of 5-bromo-2-chloro-N-(4-(4-methylpiperazin-1-yl)phenyl)pyrimidin-4-amine (22)

[0209] 2,4-Dichloro-5-bromopyrimidine (1.0 g, 4.4 mmol), 4-(4-methylpiperazin-1-yl)aniline (0.84 g, 4.4 mmol), and potassium carbonate (0.73 g, 5.3 mmol) were added to N,N-dimethylformamide (10 mL) at -20°C and allowed to react for 2 h. After completion of the reaction, water (50 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (20 mL x 3). The organic solution was collected, washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield 1.0 g of a brown oil (59.5% yield).

[0210] 4.2 Synthesis of N-benzyl-4-[(5-bromo-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidin-2-yl)amino]benzamide (23)

[0211] 5-Bromo-2-chloro-N-(4-(4-methylpiperazin-1-yl)phenyl)pyrimidin-4-amine (0.5 g, 1.3 mmol) and 4-amino-N-benzylbenzamide (0.3 g, 1.3 mmol) were added to isopropanol (10 mL) at room temperature. Trifluoroacetic acid (0.3 g, 2.6 mmol) was slowly added dropwise. After the addition was complete, the temperature was raised to 85°C and the reaction was allowed to react for 8 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and water (2 mL) was added to the residue. The pH was adjusted to 9-10 with 4 M aqueous sodium hydroxide solution, filtered, and the filter cake was washed with water and dried to obtain the crude product. The crude product was purified by preparative thin-layer chromatography to obtain 0.25 g of a light yellow solid in a yield of 33.3%.

[0212] 4.3 Synthesis of N-benzyl-4-[(4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}-5-(1H-pyrazol-4-yl)pyrimidin-2-yl)amino]benzamide

[0213] At room temperature, N-benzyl-4-[(5-bromo-4-{[4-(4-methylpiperazin-1-yl)phenyl]amino}pyrimidin-2-yl)amino]benzamide (0.2 g, 0.34 mmol), 1-H-pyrazole-4-boronate (0.08 g, 0.42 mmol), sodium carbonate (0.08 g, 0.70 mmol), and 1,1'-bis(diphenylphosphinoferrocenedichloropalladium) (0.012 g, 0.018 mmol) were added to 1,4-dioxane (5 mL). The mixture was degassed three times and heated to 100°C under a nitrogen atmosphere for 6 h. After completion of the reaction, the mixture was filtered through celite and washed with a small amount of 1,4-dioxane. The organic solution was collected and concentrated under reduced pressure to obtain the crude product, which was purified by preparative thin-layer chromatography to afford 0.06 g of a pale yellow solid in a 30.7% yield.

[0214] 4.4 Synthesis of N-benzyl-4-[(4-{[2-methoxy-4-(4-methylpiperazine-1-carbonyl)phenyl]amino}-5-(1H-pyrazol-4-yl)pyrimidin-2-yl)amino]benzamide (IV-1)

[0215] Using 2,4-dichloro-5-bromopyrimidine as the starting material, the compounds of Examples 44 and 45 were synthesized according to 4.1-1.3 of Example 43. The structures of the compounds of the examples are: 1 H-NMR data are shown in Table 4.

[0216] Table 4

[0217]

[0218]

[0219] The in vitro enzyme activity and SLP76 phosphorylation of some compounds prepared by the present invention were studied, and the results are as follows

[0220] The compounds of the 2,4-diaminopyrimidine skeleton of the general formula I of the present invention were tested for in vitro HPK1 enzyme activity using the Lantha screen assay.

[0221] Serial dilution of compounds and preparation of source plates: Dilute the compounds to 100 times the maximum inhibitor concentration required for the reaction with 100% DMSO. Transfer the compounds from the test tubes to a 96-well storage plate and serially dilute the compounds to 6 concentrations. Label this well plate as the source plate. Transfer 40μl of the dilution from the source plate to a 384-well plate as the intermediate plate. Add 5μl of kinase solution to each well of the assay plate, except for the control wells without enzyme (which add 5μl of 1x kinase buffer instead). Shake and incubate at room temperature for 10 minutes. Add 5μl 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 90 minutes.

[0222] Stop the reaction by adding 10 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 value from the Envision program. Calculate the ratio: RFU 520 nm / RFU 495 nm. Convert the ratio to percent inhibition.

[0223] Inhibition percentage = (max - sample ratio) / (max - min) * 10 ("min" represents the ratio of the no-enzyme control, and "max" represents the ratio of the DMSO control).

[0224] The data were displayed in MS Excel, and the curves were fitted by XLFit Excel version 5.4.0.8.

[0225] IC 50 The calculation formula is: Y = Bottom + (Top-Bottom) / (1 + (IC 50 / X)^HillSlope)

[0226] Table 5 IC values ​​of HPK1 kinase in some examples 50 data

[0227] Example <![CDATA[HPK1 IC 50 (nM)]]> Example <![CDATA[HPK1 IC 50 (nM)]]> Example 1 3.97 Example 20 3.16 Example 2 8.42 Example 21 0.54 Example 3 7.81 Example 22 1.05 Example 4 3.27 Example 23 1.81 Example 5 8.64 Example 24 2.88 Example 6 4.01 Example 25 2.07 Example 7 8.20 Example 26 1.85 Example 8 1.35 Example 27 1.35 Example 9 4.34 Example 28 1.08 Example 10 1.41 Example 29 0.32 Example 11 2.25 Example 30 0.59 Example 12 2.80 Example 31 1.09 Example 13 2.78 Example 32 1.68 Example 14 4.99 Example 33 4.27 Example 15 6.54 Example 34 2.25 Example 16 1.62 Example 35 2.89 Example 17 0.09 Example 40 2.16 Example 18 0.15 Example 41 1.30 Example 19 0.11 Example 43 5.61

[0228] The compounds of the general formula described in the above table all have good HPK1 inhibitory activity, and some compounds are equivalent to or better than the reference

[0229] Compounds containing the aforementioned 2,4-diaminopyrimidine backbone were further selected for SLP76 phosphorylation assays using ELISA. Jurkat cells were seeded in 90 μL of RPMI1640 medium supplemented with 10% FBS and incubated overnight at 37°C, 5% CO2. The following day, compounds were serially diluted in DMSO and added to the cells. After a 30-minute pretreatment, cells were stimulated with CD3 antibody for 15 minutes. Stimulation was stopped with ice-cold PBS, and cells were lysed with lysis buffer. Cell lysates were transferred to 96-well plates and stored at -80°C. p-SLP76 was detected using an ELISA kit. IC50 values ​​were calculated in GraphPad Prism 9.5.

[0230] IC 50 The calculation formula is: Y = Bottom + (Top-Bottom) / (1 + (IC 50 / X)^HillSlope)

[0231] Table 6 IC of SLP76 phosphorylation in some examples 50 data

[0232] Example <![CDATA[pSLP76 IC 50 (nM)]]> Example 17 133.69 Example 18 27.92 Example 19 160.82 GNE-1858 145.82

[0233] Cellular activity assays of the compounds demonstrated that the compounds effectively inhibited the phosphorylation of the HPK1 downstream adaptor protein SLP76, and compounds with superior activity compared to the control GNE-1858 were found. In vitro kinase and cell activity assays demonstrated that the compounds described herein possessed robust HPK1 enzymatic activity and excellent cellular activity, with some compounds exhibiting superior activity compared to the positive control.

Claims

1. A 2,4-diaminopyrimidine compound, characterized in that: The 2,4-diaminopyrimidine compound is a compound represented by the general formula I and a pharmaceutically acceptable salt, solvate or prodrug thereof. in, R 1 -C(O)NH2, pyrazole which is unsubstituted or substituted with at least one C1-C6 alkyl group; Y is CH, N; R 2 are independently selected from hydrogen, halogen, nitro, hydroxy, amino, cyano, (C 1- C6) alkyl, (C 1- C6) alkoxy, (C3-C 10 )cycloalkoxy; L is a single bond, -CH2, -C(O), -S(O), -S(O)2; R 3 and R 4 are the same or different, independently selected from hydrogen, (C 1- C6) alkyl, (C 3- C 10 ) cycloalkyl, 4-10 membered heterocyclic group; or R 3 and R 4 Together with the nitrogen atom to which they are attached, they form a 4-10 membered monocyclic heterocyclic group or a bridged bicyclic heterocyclic group; R 5 are independently selected from hydrogen, halogen, nitro, hydroxy, amino, cyano, (C 1- C6) alkyl, (C 1- C6) alkoxy; m is 0, 1 or 2; When m is 0, R 6 is hydrogen; R 7 Selected from (C 1- C6) alkyl, (C3-C 10 )cycloalkyl, (C6-C 10 ) aryl, 4-10 membered heterocyclic group, or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 4-10 membered heterocyclic group; When m is 1 or 2, R 6 is selected from hydrogen or methyl; R 7 Selected from hydrogen, (C3-C 10 )cycloalkyl, (C6-C 10 ) aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclyl; The R 2 、R 3 、R 4 、R 5 and R 7 (C 1- C6) alkyl, (C 1- C6) alkoxy, (C3-C 10 )cycloalkoxy, heterocyclic, aryl, heteroaryl, R 3 and R 4 The heterocyclic group formed, R 6 and R 7 The heterocyclic group is independently optionally substituted by 0-3 identical or different R 8 replace; All heterocyclic and heteroaryl groups mentioned above may optionally contain 1-4 identical or different heteroatoms selected from N, O, and S, wherein the heterocyclic group may optionally contain 0-2 carbon-carbon double bonds or carbon-carbon triple bonds; R 8 are independently selected from hydrogen, halogen, oxo, nitro, hydroxy, amino, cyano, mercapto, 4-10 membered heterocyclic groups, free, salified, esterified and amidated carboxyl, (C 1- C6) alkylsulfinyl, (C 1- C6) alkylsulfonyl, (C 1- C6) alkylacyl, unsubstituted or substituted with at least one hydroxyl, amino or halogen (C 1- C6) alkyl or (C 1- C6) alkoxy.

2. The 2,4-diaminopyrimidine compound according to claim 1, characterized in that: The compound of general formula I and its pharmaceutically acceptable salt, solvate or prodrug, in, R 1 -C(O)NH2, pyrazole which is unsubstituted or substituted with at least one C1-C6 alkyl group; Y is CH, N; R 2 Independently selected from hydrogen, halogen, nitro, hydroxy, amino, cyano, (C1-C4) alkyl, (C1-C4) alkoxy, (C3-C7) cycloalkoxy; L is a single bond, -CH2, -C(O), -S(O), -S(O)2; R 3 and R 4 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl, (C3-C7) cycloalkyl, 4-6 membered heterocyclyl; or R 3 and R 4 Together with the nitrogen atom to which they are attached, they form a 4-10 membered monocyclic heterocyclic group or a bridged bicyclic heterocyclic group; R 5 independently selected from hydrogen, halogen, nitro, hydroxy, amino, cyano, (C1-C4) alkyl, (C1-C4) alkoxy; m is 0, 1 or 2; When m is 0, R 6 is hydrogen; R 7 Selected from (C 1- C4) alkyl, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 4-6 membered heterocyclic group, or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group; When m is 1 or 2, R 6 is selected from hydrogen or methyl; R 7 selected from hydrogen, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 5-6 membered heteroaryl, 4-6 membered heterocyclyl; The R 2 、R 3 、R 4 、R 5 and R 7 (C 1- C4) alkyl, (C 1- C4)alkoxy, (C3-C7)cycloalkoxy, heterocyclic, aryl, heteroaryl R 3 and R 4 The heterocyclic group formed, R 6 and R 7 The heterocyclic group is independently optionally substituted by 0-3 identical or different R 8 replace; All heterocyclic and heteroaryl groups mentioned above may optionally contain 1-4 identical or different heteroatoms selected from N, O, and S, wherein the heterocyclic group may optionally contain 0-2 carbon-carbon double bonds or carbon-carbon triple bonds; R 8 Independently selected from hydrogen, halogen, oxo, nitro, hydroxy, amino, cyano, mercapto, 4-6 membered heterocyclic groups, free, salified, esterified and amidated carboxyl, (C1-C4) alkylsulfinyl, (C1-C4) alkylsulfonyl, (C1-C4) alkylacyl, (C1-C4) alkyl or (C1-C4) alkoxy groups which are unsubstituted or substituted by at least one hydroxy, amino or halogen group.

3. The 2,4-diaminopyrimidine compound according to claim 2, characterized in that: The compound of general formula I and its pharmaceutically acceptable salt, solvate or prodrug, in, R 1 is -C(O)NH2, 1H-pyrazol-4-yl, 1-methyl-1H-pyrazol-4-yl; Y is CH, N; R 2 independently selected from hydrogen, halogen, (C1-C4)alkyl, (C1-C4)alkoxy, (C3-C7)cycloalkoxy; L is a single bond, -CH2, or -C(O); R 3 and R 4 are the same or different and are independently selected from hydrogen, a 4-6 membered heterocyclic group; or R 3 and R 4 Together with the nitrogen atom to which they are attached, they form a 4-10 membered monocyclic heterocyclic group or a bridged bicyclic heterocyclic group; R 5 independently selected from hydrogen, halogen, (C1-C4)alkyl, (C1-C4)alkoxy; m is 0, 1 or 2; When m is 0, R 6 is hydrogen; R 7 Selected from (C 1- C4) alkyl, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 4-6 membered heterocyclic group, or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group; When m is 1 or 2, R 6 is selected from hydrogen or methyl; R 7 selected from hydrogen, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 5-6 membered heteroaryl, 4-6 membered heterocyclyl; The R 2 、R 3 、R 4 、R 5 and R 7 (C 1- C4) alkyl, (C 1- C4)alkoxy, (C3-C7)cycloalkoxy, heterocyclyl, aryl, heteroaryl, R 3 and R 4 The heterocyclic group formed, R 6 and R 7 The heterocyclic group is independently optionally substituted by 0-3 identical or different R 8 replace; All heterocyclic and heteroaryl groups mentioned above may optionally contain 1-4 identical or different heteroatoms selected from N, O, and S, wherein the heterocyclic group may optionally contain 0-2 carbon-carbon double bonds or carbon-carbon triple bonds; R 8 Independently selected from hydrogen, halogen, oxo, hydroxy, cyano, (C1-C4) alkyl, (C1-C4) alkoxy, 4-6 membered heterocyclyl, (C1-C4) alkyl or (C1-C4) alkoxy substituted by at least one halogen.

4. The 2,4-diaminopyrimidine compound according to claim 3, characterized in that: The compound of general formula I and its pharmaceutically acceptable salt, solvate or prodrug, in, R 1 is -C(O)NH2; Y is CH; R 2 independently selected from hydrogen, halogen, methyl, methoxy, ethoxy; L is a single bond, -CH2, or -C(O); R 3 and R 4 Together with the nitrogen atom to which they are attached, they form a 4-10 membered monocyclic heterocyclic group or a bridged bicyclic heterocyclic group; R 5 independently selected from hydrogen, halogen, methyl, methoxy, ethoxy; m is 0, 1 or 2; When m is 0, R 6 is hydrogen; R 7 Selected from (C 1- C4) alkyl, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 4-6 membered heterocyclic group, or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group; When m is 1 or 2, R 6 is selected from hydrogen; R 7 selected from hydrogen, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 5-6 membered heteroaryl, 4-6 membered heterocyclyl; The R 3 、R 4 and R 7 Heterocyclic group, aryl group, heteroaryl group, R 3 and R 4 The heterocyclic group is independently optionally substituted by 0-3 identical or different R 8 replace; All heterocyclic and heteroaryl groups mentioned above may optionally contain 1-4 identical or different heteroatoms selected from N, O, and S, wherein the heterocyclic group may optionally contain 0-2 carbon-carbon double bonds or carbon-carbon triple bonds; R 8 Independently selected from hydrogen, halogen, oxo, hydroxy, cyano, methyl, methoxy, trifluoromethyl.

5. The 2,4-diaminopyrimidine compound according to claim 4, characterized in that: The compound of general formula I and its pharmaceutically acceptable salt, solvate or prodrug, in, R 1 is -C(O)NH2; Y is CH; R 2 independently selected from hydrogen, halogen, methyl, methoxy, ethoxy; L is a single bond, -CH2, or -C(O); R 3 and R 4 Together with the nitrogen atoms to which they are attached, they form: R 5 independently selected from hydrogen, halogen, methyl, methoxy, ethoxy; m is 0 or 1; When m is 0, R 6 is hydrogen; R 7 Selected from (C 1- C4) alkyl, (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 4-6 membered heterocyclic group, or R 6 and R 7 Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group; When m is 1, R 6 is selected from hydrogen; R 7 Selected from (C3-C7) cycloalkyl, (C6-C 10 ) aryl, 5-6 membered heteroaryl, 4-6 membered heterocyclyl; The R 7 The heterocyclic group, aryl group and heteroaryl group in the 8 replace; All heterocyclic and heteroaryl groups mentioned above may optionally contain 1-4 identical or different heteroatoms selected from N, O, and S, wherein the heterocyclic group may optionally contain 0-2 carbon-carbon double bonds or carbon-carbon triple bonds; R 8 Independently selected from hydrogen, halogen, oxo, hydroxy, cyano, methyl, methoxy, trifluoromethyl.

6. The 2,4-diaminopyrimidine compound according to any one of claims 1 to 5, characterized in that: The salt of the compound of general formula I is a salt formed by the compound of general formula I and 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.

7. A pharmaceutical composition, characterized in that: The composition contains the compound of general formula I according to any one of claims 1 to 5 and its pharmaceutically acceptable salt, hydrate, solvate or prodrug.

8. Use of the compound according to claim 1, characterized in that: Use of the compound according to any one of claims 1 to 5 and pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof in the preparation of drugs for preventing or treating diseases associated with HPK1 kinase.

9. The use according to claim 8, characterized in that: The diseases mediated by HPK1 are prevented from viral infection, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, gastric cancer, skin cancer, ovarian cancer, cervical cancer, kidney cancer, head and neck cancer, lymphoma, leukemia, and melanoma.

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