A benzimidazole derivative and use thereof

Abstract

The present application relates to benzimidazole derivatives containing dithiocarbamic acid structure of general formula I and their optical isomers, pharmaceutically acceptable salts, solvates or prodrugs, their preparation method and pharmaceutical compositions of the compounds of general formula I as active ingredients, wherein the substituents R1, R2, A, L, X have the meanings given in the specification. The present application also relates to the use of the compounds of general formula I and their pharmaceutically acceptable salts, solvates or prodrugs thereof for the preparation of a medicament for the treatment of diseases caused by the abnormally high expression of PI3K kinases, in particular for the preparation of a medicament for the treatment and / or prevention of cancer.

Description

Technical Field

[0001] This invention relates to pharmaceutical compounds, specifically to benzimidazole derivatives containing a dithiocarbamate structure and their use in the preparation of medicaments for treating and / or preventing diseases caused by abnormally high expression of PI3K, particularly in the preparation of medicaments for treating and / or preventing cancer. Background Technology

[0002] Cancer, encompassing all malignant tumors, is one of the two leading causes of death worldwide, alongside cardiovascular and cerebrovascular diseases. It is a major disease seriously affecting human health and threatening human life. Physical, chemical, and viral factors are all major contributing factors to cancer. Cancer cells grow rapidly, invading surrounding tissues and spreading, exhibiting powerful destructive capabilities. Besides causing obstructive and compressive symptoms, they can also affect the normal function of organs and genes, leading to death. With advancements in molecular biology techniques, the pathogenesis of tumors has been further understood at the level of cell receptors and proliferation regulatory molecules, leading to the increasing maturity of targeted therapies targeting cell receptors and proliferation regulatory molecules. Among these, targeted drugs, due to their precision, high efficacy, significant therapeutic effects, and significantly lower toxicity compared to chemotherapy, along with ease of use, have gradually gained popularity and become a hot topic in modern anti-cancer drugs.

[0003] Phosphatidylinositol-3-kinase (PI3K), a lipid kinase located in a cytoplasmic complex, catalyzes the phosphorylation of phosphatidylinositol at the D3 position. PI3K plays a crucial role in cell proliferation, growth, and survival. When cells are stimulated by various growth factors, activation of PI3K leads to the conversion of phosphatidylinositol bisphosphate (PIP2) to phosphatidylinositol triphosphate (PIP3). PIP3, acting as a second messenger, then reactivates numerous downstream proteins.

[0004] PI3K is classified into three types—I, II, and III—based on its structure and phosphorylation substrate. Type I PI3K consists of a regulatory subunit p85 and a catalytic subunit p110, and generally catalyzes the phosphorylation of PIP2 to generate PIP3. Because type I PI3K has been the most extensively studied functionally to date, it is commonly referred to as PI3K. Type I PI3K is further divided into four subtypes based on differences in their catalytic subunits, etc.: PI3Kα, PI3Kβ, PI3Kδ, and PI3Kγ. These four subtypes also differ in function. Reported PI3K gene mutations in cancer patients are generally p110α mutations; therefore, PI3Kα is considered to play an important role in tumorigenesis.

[0005] Currently, with the continuous development of medicinal chemistry, oncology, and pharmacology, global research on PI3K-targeted drugs has achieved certain results. Several PI3K inhibitors are currently in clinical trials worldwide, among which ZSTK-474 is a pan-PI3K inhibitor developed by Zenyaku Kogyo in collaboration with the Japanese Cancer Research Society. Similar to most other PI3K inhibitors, it induces cell cycle arrest in the G1 phase in various tumor cells without significant apoptosis. Its cell cycle arrest mechanism may be attributed to the inactivation of cyclin D1, increased p27 expression, and pRB dephosphorylation. ZSTK-474 (Fig. 1) shows the IC50 values ​​for PI3Kα, β, δ, and γ. 50 The concentrations were 16, 44, 5, and 49 nmol / L, respectively. Oral administration of ZSTK-474 showed good anti-tumor effects in various nude mouse xenograft models and is currently undergoing Phase I clinical trials for evaluation.

[0006]

[0007] However, the drugs developed at this stage have drawbacks such as weak inhibitory effect on the target and low selectivity, which limit their clinical use. Summary of the Invention

[0008] The present invention aims to provide a benzimidazole derivative and its use in the preparation of medicaments for treating and / or preventing diseases caused by abnormally high expression of PI3K, particularly in the preparation of medicaments for treating and / or preventing cancer.

[0009] To achieve the above objectives, the present invention adopts the following technical solution:

[0010] A benzimidazole derivative: the derivative is a benzimidazole derivative containing a dithiocarbamate structure, having the structure shown in general formula I, and its stereoisomers, pharmaceutically acceptable salts, solvates, or prodrugs.

[0011]

[0012] in,

[0013] X represents N and CH;

[0014] R1 is hydrogen, an unsubstituted (C1-C6)alkoxy group, or a (C3-C6)cycloalkoxy group substituted with at least one of the following groups: hydroxyl, amino, or dimethylamino.

[0015] Ring A is selected from

[0016] L is

[0017] Y is C=O, CH2;

[0018] When L is L1, R2 is -NR3R4; when L is L2, R2 is -NR3R4, cyano, alkenyl, hydroxyl, amino, carboxyl, (C1–C6)alkoxy, (C1–C6)alkoxyformyl, (C1–C6)alkylsulfonyl, (C6–C 10 ) aryl, 5-10 heteroaryl, wherein the aryl or heteroaryl is optionally replaced by 1-3 identical or different R5;

[0019] R5 is a hydroxyl, halogen, nitro, amino, cyano, (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)alkyl or (C1-C6)alkoxy optionally substituted with hydroxyl, amino or halogen, amino substituted with 1-2 (C1-C6)alkyl, (C1-C6)alkylamide, free or salt-forming or esterified or amidated carboxyl, (C1-C6)alkylsulfinyl, (C1-C6)alkylsulfonyl, (C1-C6)alkylacyl, carbamoyl, (C1-C3)alkyldioxy, (C6-C 10 )Aryl, (C3-C 10 )cycloalkyl, 5-10-membered heteroaryl, 4-10-membered heterocyclic;

[0020] R3 and R4 may be the same or different, and are independently selected from hydrogen, (C2-C6) alkenyl, (C2-C6) alkynyl, unsubstituted or substituted with 1-3 identical or different R6, (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl, and dimethylaminoethyl;

[0021] Alternatively, R3 and R4 together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclic group containing at least one heteroatom; the heterocyclic group optionally contains 1-4 heteroatoms selected from N, O and / or S, the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, or the heterocyclic group is optionally substituted with one or more oxygen atoms, and the heterocyclic group is optionally substituted with 0-3 identical or different R7 atoms;

[0022] R6 and R7 may be the same or different, and are independently selected from hydrogen, hydroxyl, halogen, amino group substituted with 0-2 (C1-C6) alkyl groups, cyano group, (C1-C6) alkyl acyl group, (C1-C6) alkyl sulfonyl group, unsubstituted (C1-C6) alkyl group or substituted with at least one of the following groups, (C1-C6) alkoxy group, (C3-C6) cycloalkyl group; the following groups are hydroxyl, amino or halogen;

[0023] n is an integer between 1 and 2;

[0024] m is an integer between 0 and 4.

[0025] Preferably, the derivative is a compound of general formula I and its stereoisomers, a pharmaceutically acceptable salt, a solvate, or a prodrug thereof, wherein,

[0026] X represents N and CH;

[0027] R1 is a hydrogen, an unsubstituted (C1-C4) alkoxy group, or a group substituted with at least one of the following groups: hydroxyl, amino, or dimethylamino.

[0028] Ring A is selected from

[0029] L is

[0030] Y is C=O, CH2;

[0031] When L is L1, R2 is -NR3R4; when L is L2, R2 is -NR3R4, cyano, alkenyl, hydroxyl, amino, carboxyl, (C1–C4)alkoxy, (C1–C4)alkoxyformyl, (C1–C4)alkylsulfonyl, (C6–C4)alkylsulfonyl, (C6–C4)alkylsulfonyl, (C1 ... 10 ) aryl, 5-6 heteroaryl, wherein the aryl or heteroaryl is optionally substituted by 1-3 identical or different R5;

[0032] R5 is a hydroxyl, halogen, nitro, amino, cyano, (C1-C4)alkyl, (C1-C4)alkoxy, (C1-C4)alkyl or (C1-C4)alkoxy optionally substituted with hydroxyl, amino or halogen, amino substituted with 1-2 (C1-C4)alkyl, (C1-C4)alkylamide, free or salt-forming or esterified or amidated carboxyl, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, (C1-C4)alkylacyl, carbamoyl, (C1-C3)alkyldioxy, (C6-C 10 )Aryl, (C3-C 10 )cycloalkyl, 5-6 membered heteroaryl, 4-6 membered heterocyclic;

[0033] R3 and R4 may be the same or different, and are independently selected from hydrogen, dimethylaminoethyl, unsubstituted or substituted with 1-3 identical or different R6 (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl;

[0034] Alternatively, R3 and R4 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing at least one heteroatom; the heterocyclic group optionally contains 1-4 heteroatoms selected from N, O and / or S, the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, or the heterocyclic group is optionally substituted with one or more oxygen atoms, and the heterocyclic group is optionally substituted with 0-3 identical or different R7 atoms;

[0035] R6 and R7 may be the same or different, and are independently selected from hydrogen, hydroxyl, halogen, amino group substituted with 0-2 (C1-C4) alkyl groups, cyano group, (C1-C4) alkyl acyl group, (C1-C4) alkyl sulfonyl group, unsubstituted (C1-C4) alkyl group or substituted with at least one of the following groups, (C1-C4) alkoxy group, (C3-C6) cycloalkyl group; the following groups are hydroxyl, amino or halogen;

[0036] n is an integer between 1 and 2;

[0037] m is an integer between 0 and 4.

[0038] In a further preferred embodiment, the derivative is a compound of general formula I and its stereoisomers, a pharmaceutically acceptable salt, a solvate, or a prodrug thereof.

[0039] X represents N and CH;

[0040] R1 is hydrogen or (C1-C4)alkoxy;

[0041] Ring A is selected from

[0042] L is

[0043] Y is C=O, CH2;

[0044] When L is L1, R2 is -NR3R4; when L is L2, R2 is -NR3R4, cyano, alkenyl, hydroxyl, amino, carboxyl, (C1–C4)alkoxy, (C1–C4)alkoxyformyl, (C1–C4)alkylsulfonyl.

[0045] R3 and R4 may be the same or different, and are independently selected from hydrogen, dimethylaminoethyl, unsubstituted or substituted with 1-3 identical or different R6 (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl;

[0046] Alternatively, R3 and R4 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing at least one heteroatom; the heterocyclic group optionally contains 1-4 heteroatoms selected from N, O and / or S, the heterocyclic group contains 0-2 carbon-carbon double bonds or carbon-carbon triple bonds, or the heterocyclic group is optionally substituted with one or more oxygen atoms, and the heterocyclic group is optionally substituted with 0-3 identical or different R7 atoms;

[0047] R6 and R7 may be the same or different, and are independently selected from hydrogen, hydroxyl, halogen, amino group substituted with 0-2 (C1-C4) alkyl groups, cyano group, (C1-C4) alkyl acyl group, (C1-C4) alkyl sulfonyl group, unsubstituted (C1-C4) alkyl group or substituted with at least one of the following groups, (C1-C4) alkoxy group, (C3-C6) cycloalkyl group; the following groups are hydroxyl, amino or halogen;

[0048] n is an integer between 1 and 2;

[0049] m is an integer between 1 and 2.

[0050] Further preferably, the derivative is a compound of general formula I and its stereoisomers, a pharmaceutically acceptable salt, a solvate, or a prodrug thereof, wherein,

[0051] X represents N and CH;

[0052] R1 is hydrogen or methoxy;

[0053] Ring A is selected from

[0054] L is

[0055] Y is C=O, CH2;

[0056] When L is L1, R2 is -NR3R4; when L is L2, R2 is -NR3R4, cyano, alkenyl, hydroxyl, amino, carboxyl, (C1–C4)alkoxy, (C1–C4)alkoxyformyl, (C1–C4)alkylsulfonyl.

[0057] R3 and R4 may be the same or different, and are independently selected from hydrogen, dimethylaminoethyl, unsubstituted or substituted with 1-3 identical or different R6 (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl;

[0058] Or R3 and R4 together with the nitrogen atoms they are attached to form

[0059] n is an integer between 1 and 2;

[0060] m is an integer between 1 and 2.

[0061] More preferably, the derivative is a compound of general formula I and its stereoisomers, a pharmaceutically acceptable salt, a solvate, or a prodrug thereof.

[0062] 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyldiethyldithiocarbamate;

[0063] 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-yl}-2-oxoethylpyrrolidine-1-carbodisulfate;

[0064] 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-yl}-2-oxoethylmorpholino-4-carbodisulfate;

[0065] 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-yl}-2-oxoethylpiperidine-1-carbodisulfate;

[0066] 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazin-1-yl}-2-oxoethyl 4-methylpiperazine-1-carbodisulfate;

[0067] 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 4-(methanesulfonyl)piperazin-1-carbodisulfate;

[0068] 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 4-(2-hydroxyethyl)piperazin-1-carbodisulfate;

[0069] 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazin-1-yl}-2-oxoethyl-4-ethylpiperazine-1-carbodisulfate;

[0070] 2-(dimethylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0071] 2-(diethylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0072] 2-morpholino-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0073] 2-(4-methylpiperazin-1-yl)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate;

[0074] 2-Oxo-2-(pyrrolidone-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate;

[0075] 2-(4-(methanesulfonyl)piperazin-1-yl)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate;

[0076] 2-(4-Cyclopentylpiperazin-1-yl)-oxyethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate;

[0077] 2-(cyclobutanylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0078] 2-Oxo-2-(piperidin-1-yl)ethyl(S)-{1-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}pyrrolidine-3-yl}aminodithiocarbamate;

[0079] 2-Oxo-2-(piperidin-1-yl)ethyl{1-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperidin-4-yl}aminodithiocarbamate;

[0080] 2-Methoxyethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0081] 2-(dimethylaminoethyl)4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0082] 2-(4-methylpiperazin-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate;

[0083] 2-Morpholinyl ethyl ester 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0084] 2-(piperidin-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate;

[0085] 2-(pyrrolidone-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0086] Pyridin-4-ylmethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0087] cyanomethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0088] 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}-1,4-diaza-1-yl}2-oxoethylmorpholino-4-dithiocarbamate;

[0089] (S)-2-{3-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}amino}pyrrolidine-1-yl}-2-oxoethylmorpholino-4-dithiocarbamate;

[0090] (S)-2-{3-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}amino}pyrrolidine-1-yl}-2-oxoethyl 4-(2-hydroxyethyl)piperazine-1-carbodisulfate;

[0091] 2-{4-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}amino}piperidin-1-yl}-2-oxoethylmorpholino-4-dithiocarbamate;

[0092] 2-{4-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}amino}piperidin-1-yl}-2-oxoethyl 4-(2-hydroxyethyl)piperazine-1-carbodisulfate;

[0093] 2-{4-{4-[2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylmorpholino-4-carbodisulfate;

[0094] 2-{4-{4-[2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 4-ethylpiperazin-1-carbodisulfate;

[0095] 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethyl diethyl dithiocarbamate;

[0096] 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylpyrrolidine-1-carbodisulfate;

[0097] 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-4-carbodisulfate;

[0098] 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylpiperidine-1-carbodisulfate;

[0099] 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethyl 4-(methanesulfonyl)piperazin-1-carbodisulfate;

[0100] 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-1,1-dioxide-4-carbodisulfate;

[0101] 2-(dimethylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0102] 2-(diethylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0103] 2-O-2-(pyrrolidone-1-yl)ethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0104] 2-morpholino-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0105] 2-(cyclobutanylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0106] 2-Methoxyethyl 4-{2-[2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0107] 2-(dimethylamino)4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0108] 2-(pyrrolidone-1-yl)ethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0109] Pyridin-4-ylmethyl 4-{2-[2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0110] cyanomethyl 4-{2-[2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0111] 2-Amino-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0112] 2-(cyclohexylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0113] 2-(cyclopentylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0114] 2-(cyclopropylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate;

[0115] 2-(cyclopropylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0116] 2-Amino-2-oxoethyl 4-{2-[2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate;

[0117] 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-4-dithiocarbamate;

[0118] 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylcyclopropylaminodithiocarbamate;

[0119] 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 3,5-dimethylpiperazin-1-dithiocarbamate;

[0120] 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-4-dithiocarboxylic acid 1,1-dioxide;

[0121] 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethyl-4-cyclopentylpiperazin-1-dithiocarbamate.

[0122] The compound salt 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.

[0123] Furthermore, the present invention also includes prodrugs derived from the present invention. These prodrugs are derivatives of general formula I, which may themselves have weak or even no activity, but are converted into their corresponding biologically active forms under physiological conditions (e.g., through metabolism, solvation, or other means) after administration.

[0124] In this invention, "halogen" refers to fluorine, chlorine, bromine, or iodide; "alkyl" refers to a straight-chain or branched alkyl group; "alkylene" refers to a straight-chain or branched alkylene group; "cycloalkyl" refers to a substituted or unsubstituted cycloalkyl group; "aryl" refers to an unsubstituted or substituted phenyl or naphthyl group; "heteroaryl" refers to a cyclic system containing one or more monocyclic or polycyclic heteroatoms selected from N, O, and S, wherein the cyclic system is aromatic, such as imidazolyl, pyridinyl, pyrazolyl, etc. 1,2,3)- and (1,2,4)-triazolyl, furanyl, thiopheneyl, pyrroleyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl, quinolinyl, isoquinolinyl, benzimidazolyl, and benzoxazolyl, etc.; "saturated or partially saturated heterocyclic groups" refer to cyclic systems containing one or more heteroatoms selected from N, O, and S, such as pyrroleyl, morpholinyl, piperazineyl, piperidinyl, pyrazolylyl, imidazolyl, and thiazolyl, etc.

[0125] When selecting the above substituents, For example, when n is 1, it is a six-membered ring containing heteroatoms; when n is 2, it is a seven-membered ring containing heteroatoms, and so on.

[0126] In the above substituents, taking L1 as an example, when m is 0, Y and S are directly connected; when m is 1, Y and S are connected through a methylene group, and so on.

[0127] A pharmaceutical composition comprising a compound of formula I and its stereoisomers, a pharmaceutically acceptable salt, a solvate or a prodrug thereof, mixed with a pharmaceutically acceptable carrier.

[0128] The carriers used in the pharmaceutical compositions of this invention are common types available in the pharmaceutical field, including: binders, lubricants, disintegrants, solubilizers, diluents, stabilizers, suspending agents, colorants, flavoring agents, etc., for oral formulations; preservatives, solubilizers, stabilizers, etc., for injectable formulations; and matrices, diluents, lubricants, preservatives, etc., for topical formulations. The pharmaceutical formulations can be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically). If certain drugs are unstable under gastric conditions, they can be formulated into enteric-coated tablets.

[0129] The use of a compound and a pharmaceutical composition thereof, wherein the compound of general formula I and its stereoisomers, pharmaceutically acceptable salts, solvates or prodrugs thereof, or a pharmaceutical composition containing a compound of general formula I and its stereoisomers, pharmaceutically acceptable salts, solvates or prodrugs thereof, in the preparation of a medicament for the treatment and / or prevention of PI3K-mediated diseases.

[0130] The diseases that are prevented from being PI3K mediated include lung cancer, liver cancer, stomach cancer, colorectal cancer, breast cancer, glioblastoma, bladder cancer, prostate cancer, ovarian cancer, or esophageal cancer.

[0131] The PI3K activity test revealed that the compound of the present invention has a significant inhibitory effect on PI3K activity and a strong inhibitory effect on lung cancer cells, colon cancer cells, gastric cancer and breast cancer cells with high PI3K expression. It is particularly useful for the preparation of drugs for the treatment and / or prevention of lung cancer.

[0132] By inhibiting human colon cancer cells HCT-116, human bladder cancer cells PC-3, and human breast cancer cells MCF-7 in vitro, we found that the compounds of the present invention have significant anti-tumor activity. Therefore, the compounds of the present invention can be used to prepare drugs for the treatment or prevention of lung cancer cells, prostate cancer cells, and malignant glioblastoma.

[0133] The precise amount of the compounds of this invention required for treating malignant tumors will vary from person to person, depending on the type of patient, age and general condition, severity of the disease being treated, the specific compound used, and the route of administration, such as the route and frequency of administration. Those skilled in the art can determine the appropriate effective amount using only conventional experimental methods.

[0134] The dosage of the compound can range from 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 or hematologic malignancy being treated, and the specific compound used. Furthermore, it is understood that the initial dose may be increased beyond the upper limit to rapidly achieve the desired blood levels, or the initial dose may be lower than the optimal value, and the daily dose may be gradually increased during treatment, depending on the specific circumstances. If necessary, the daily dose may also be divided into multiple doses, for example, 2-4 times daily.

[0135] Mammals refer to humans or animals.

[0136] The amount of the active ingredient, i.e., the compound according to the 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 required concentration. Generally, the content of the active ingredient will be between 0.5% and 90% by total weight of the composition.

[0137] In combination therapy, the compounds of the present invention and other compounds may be administered simultaneously or at intervals. When administered simultaneously, the compounds of the present invention and other compounds may be combined in a single pharmaceutical composition or in separate compositions.

[0138] The examples and preparation methods provided below further illustrate and demonstrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparation methods does not limit the scope of the present invention in any way.

[0139] The following synthetic routes (Route 1, Route 2, Route 3, and Route 4) describe the preparation of the general formula I derivatives of this invention. All starting materials were 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 this invention were prepared by the methods described in these synthetic routes or by similar methods well known to those skilled in the art of organic chemistry. All variable factors used in these synthetic routes are defined below or as defined in the claims.

[0140] That is, depending on the choice of ring A, the general formula compound can be prepared according to the following different routes;

[0141]

[0142]

[0143] According to the derivatives of Formula I of the present invention, T1, T3, T5 and T7 can be prepared by reacting the corresponding intermediates I-1, I-2 or I-3 with carbon disulfide and sodium hydroxide for 4 hours according to routes 1, 2, 3 and 4, to obtain sodium salts, and then reacting them with small molecule amines substituted with haloacyl chlorides through a substitution reaction; T2, T4, T6 and T8 can be prepared by reacting the corresponding intermediates I-1, I-2 or I-3 with haloacyl chlorides through a substitution reaction, and then reacting them with substituted dithiocarbamates under alkaline conditions.

[0144] When ring A is The preparation method of intermediate I-1 is shown in route 5:

[0145]

[0146] When ring A is The preparation method of intermediate I-2 is shown in route 6:

[0147]

[0148] When ring A is The preparation method of intermediate I-3 is shown in route 7:

[0149]

[0150] The substituents R1, R3, R4, m, n, X, etc. of all intermediates in the above 7 routes are as defined in the claims.

[0151] Advantages of this invention:

[0152] The series of benzimidazole derivatives designed and synthesized in this invention mainly involve the introduction of a dithiocarbamate structure with antitumor activity at a specific position in their structure for the first time. Pharmacological activity tests have shown that they have good antitumor activity and significant inhibitory activity against PI3K, while also showing significant inhibitory effects on various human tumor cells. Detailed Implementation

[0153] The following examples further illustrate specific embodiments of the present invention. It should be noted that the specific embodiments described herein are merely for illustration and explanation and are not intended to limit the scope of the present invention.

[0154] The examples are intended to illustrate, and not limit, the scope of the invention. The proton NMR spectra of the compounds were determined using a Bruker ARX-400 or Bruker ARX-600, and the mass spectrometry was performed using an Agilent 1100 LC / MSD; all reagents used were analytical grade or chemically pure.

[0155]

[0156]

[0157]

[0158]

[0159] Preparation of Example 1:

[0160]

[0161] Step A2: Preparation of 5-methoxy-nitroaniline (A1)

[0162] 2-Amino-3-nitrobenzene ether (30.0 g, 178.6 mmol) and Pd / C (5.7 g, 5% w / w) were added to 250 mL of methanol at room temperature. The reaction mixture was heated to 50 °C for 4 h under H₂ protection. After the reaction was complete, the mixture was cooled to room temperature, filtered, and Pd / C was recovered. The filtrate was collected, and the solvent was removed by vacuum evaporation to obtain 23.3 g of a brown powdery solid, with a yield of 94.5%. MS (ESI) m / z: 139.0 [M+H] + .

[0163] Step B: Preparation of 2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazole (A2)

[0164] At room temperature, intermediate A1 (20.0 g, 144.9 mmol) and difluoroacetic acid (13.9 g, 144.9 mmol) were added to 100 mL of PPA, and the reaction solution was heated to 120 °C and reacted for 5 h. After the reaction was complete, the reaction solution was cooled to room temperature and then poured into 200 mL of ice water. The pH was adjusted to approximately 7 with 10% sodium hydroxide solution, and a brown solid precipitated. The solid was filtered, and the filter cake was washed with 50 mL of water. After vacuum drying, 26.2 g of brown powdery solid was obtained, with a yield of 91.2%. MS (ESI) m / z: 199.1 [M+H] + .

[0165] Step C 4-(4,6-dichloro-1,3,5-triazin-2-yl)morpholine (4)

[0166] At room temperature, cyanuric chloride (50.0 g, 695.5 mmol) and sodium bicarbonate (50.0 g, 695.5 mmol) were added to 500 mL of acetone. The temperature was lowered to -20 °C, and morpholine (50.0 g, 695.5 mmol) was slowly added in portions. After the addition was complete, the reaction was maintained at -20 °C for 1 h, resulting in the precipitation of a white solid. After the reaction was complete, the reaction solution was filtered, and the filter cake was washed with a small amount of acetone. The filter cake was dried to obtain a white solid with a yield of 81.2%. MS (ESI) m / z: 235.1 [M+H] + .

[0167] Step D: Preparation of 4-{4-chloro-6-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-1,3,5-triazin-2-yl}morpholine (A3)

[0168] At room temperature, intermediate 4 (20.0 g, 85.1 mmol) and potassium carbonate (22.1 g, 102.1 mmol) were added to 120 mL of LDM. Intermediate A2 (15.8 g, 85.1 mmol) was added in portions to the reaction mixture. After the addition was complete, the mixture was allowed to react at room temperature for 4 h. After the reaction was complete, the reaction mixture was poured into 200 mL of water, and a solid precipitated. The solid was filtered, and the filter cake was washed with 80 mL of water. The solid was dried under vacuum to give 28.7 g of a yellow powder. The yield was 85.1%. MS (ESI) m / z: 396.8 [M+H] + Preparation of step E: tert-butyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-carboxylic acid ester (A4)

[0169] At room temperature, intermediate A3 (25.0 g, 63.0 mmol), N-Boc-piperazine (17.9 g, 75.6 mmol), and DIPEA (31.0 g, 126.0 mmol) were added sequentially to 150 mL of THF, and the reaction was carried out at room temperature for 8 h. After the reaction was complete, the reaction solution was poured into 300 mL of water, stirred for 15 min, filtered, and the filter cake was washed with 80 mL of water. The solution was dried under vacuum to obtain 31.2 g of a yellow powdery solid, with a yield of 90.5%. MS (ESI) m / z: 547.0 [M+H] + .

[0170] Preparation of step F 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-(piperazin-1-yl)-1,3,5-triazin-2-yl}morpholine (A5)

[0171] Under ice-water bath conditions, TFA (30 mL) was slowly added dropwise to a DCM (150 mL) solution of intermediate A4 (20 g, 36.6 mmol). After the addition was complete, the reaction mixture was moved to room temperature and reacted for 2 h. Upon completion of the reaction, the mixture was poured into 300 mL of ice water, and the pH was adjusted to approximately 7 with saturated NaHCO3 solution. A gray solid precipitated, which was filtered, and the filter cake was washed with 50 mL of water. Vacuum drying yielded 15.5 g of a grayish-white solid powder. The yield was 95.0%. MS (ESI) m / z: 447.1 [M+H] + 445.2 [MH] - .

[0172] Preparation of step G: 2-chloro-1-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}ethyl-1-one (A6)

[0173] Under ice-water bath conditions, A5 (20.0 g, 45.2 mmol) and DIPEA (11.6 g, 90.4 mmol) were added to 150 mL of DCM. Chloroacetyl chloride (6.0 g, 45.2 mmol) solution diluted with 20 mL of DCM was slowly added dropwise to the reaction mixture. After the addition was complete, the reaction mixture was brought to room temperature and reacted for 2 h. Upon completion of the reaction, the reaction mixture was quenched in 150 mL of water. After quenching, the mixture was extracted with DCM (50 mL × 3). The organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain 20.2 g of a pale yellow powdery solid, with a yield of 86.2%. MS (ESI) m / z: 522.9 [M+H] + 520.9 [MH] - .

[0174] Preparation of step H 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl diethyl dithiocarbamate (Example 1)

[0175] At room temperature, intermediate A6 (0.10 g, 0.19 mmol) and sodium N,N-diethylaminodithiocarbamate (0.11 g, 0.23 mmol) were dissolved in DMF (5 mL). The reaction solution was heated to 50 °C and reacted for 6–8 h. After the reaction was complete, the reaction solution was poured into water (30 mL), and a solid precipitated. The solid was filtered, washed with water, and purified by column chromatography to obtain a white solid with a yield of 65.1%; MS (ESI) m / z: 635.9 [M+H]. + 657.8 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ7.90(d,J=8.0Hz,1H),7.71(t,J=52.9Hz,1H),7.41(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.36(s,2H),3. 97(s,3H),3.97–3.85(m,4H),3.85–3.77(m,8H),3.76–3.66(m,6H),3.65–3.57(m,2H),1.26(t,J=7.1Hz,3H),1.18(t,J=7.0Hz,3H).

[0176] Following the method of Example 1, Examples 2-8 were prepared by reacting intermediate A6 as a raw material with different substituted dithiocarbamates.

[0177] Example 2: 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylpyrrolidine-1-carbodisulfate; MS (ESI) m / z: 656.7 [M+Na] + ; 1H NMR (600MHz, DMSO-d6) δ7.91(d,J=7.8Hz,1H),7.71(t,1H),7.41(t,J=8.2Hz,1H),6.95(d,J=8.1Hz,1H),4.37(s,2H),3.97(s, 3H),3.95–3.89(m,2H),3.85–3.75(m,8H),3.74–3.64(m,8H),3.63–3.56(m,2H),2.04(p,J=6.9Hz,2H),1.92(p,J=6.9Hz,2H).

[0178] Example 3: 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylmorpholino-4-carbodisulfate; MS (ESI) m / z: 649.8 [M+H] + 671.8 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ7.91(d,J=7.4Hz,1H),7.82–7.59(m,1H),7.41(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.41( s,2H),4.35–3.99(m,4H),3.97(s,3H),3.95–3.90(m,2H),3.86–3.78(m,6H),3.75–3.66(m,10H),3.64–3.58(m,2H).

[0179] Example 4: 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylpiperidine-1-carbodisulfate; MS (ESI) m / z: 647.8 [M+H] + 669.8 [M+Na] + ; 1H NMR (600MHz, DMSO-d6) δ7.90(d,J=7.7Hz,1H),7.71(t,J=52.7Hz,1H),7.41(t,J=8.2Hz,1H),6.95(d,1H),4.38(s,2H),4.25–4.18(m ,2H),3.97(s,3H),3.96–3.85(m,4H),3.84–3.78(m,6H),3.76–3.65(m,6H),3.65–3.56(m,2H),1.70–1.64(m,2H),1.62–1.52(m,4H).

[0180] Example 5: 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 4-methylpiperazin-1-carbodisulfate; MS (ESI) m / z: 662.8 [M+H] + 684.8 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ7.91(d,J=7.7Hz,1H),7.71(t,J=53.6Hz,1H),7.41(t,J=8.2Hz,1H),6.95(d,J=8.1Hz,1H),4.39(s,2 H),4.30–4.12(m,2H),3.97(s,3H),3.94–3.91(m,2H),3.89–3.64(m,14H),3.64–3.57(m,2H),2.46–2.37(m,4H),2.22(s,3H).

[0181] Example 6: 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 4-(methanesulfonyl)piperazin-1-carbodisulfate; MS (ESI) m / z: 727.1 [M+H] + 749.1 [M+Na] + ; 1H NMR (600MHz, DMSO-d6) δ7.91(d,J=7.3Hz,1H),7.71(t,J=52.9Hz,1H),7.41(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.42(s,2H),4.39– 4.04(m,4H),3.97(s,3H),3.96–3.91(m,2H),3.87–3.78(m,6H),3.77–3.67(m,6H),3.64–3.57(m,2H),3.31–3.23(m,4H),2.94(s,3H).

[0182] Example 7: 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 4-(2-hydroxyethyl)piperazin-1-carbodisulfate; MS (ESI) m / z: 693.2 [M+H] + 714.9 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ7.91(d,J=7.5Hz,1H),7.71(t,J=52.7Hz,1H),7.41(t,J =8.2Hz,1H),6.95(d,J=8.1Hz,1H),4.48(t,J=5.2Hz,1H),4.39(s,2H),4.23(d,J =21.1Hz,2H),3.97(s,3H),3.97–3.87(m,4H),3.86–3.78(m,6H),3.76–3.66(m,6 H),3.65–3.57(m,2H),3.52(q,J=5.9Hz,2H),2.52(s,4H),2.44(t,J=6.0Hz,2H);

[0183] Example 8: 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl-4-ethylpiperazin-1-carbodisulfate; MS (ESI) m / z: 677.0 [M+H] + 698.9 [M+Na] + ; 1H NMR (600MHz, DMSO-d6) δ7.91(d,J=7.6Hz,1H),7.71(t,J=52.9Hz,1H),7.41(t,J=8.2Hz,1H),6.95(d,1H),4.39(s,2H),4.30–4.12(m,2H),3.97( s,3H),3.97–3.87(m,4H),3.85–3.78(m,6H),3.76–3.67(m,6H),3.64–3 .57(m,2H),2.49–2.41(m,4H),2.40–2.34(m,2H),1.02(t,J=7.2Hz,3H).

[0184] Preparation of Example 9:

[0185] Step I: Preparation of sodium 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-carbodithiosulfate (A7)

[0186] At room temperature, intermediate A6 (36.7 g, 234.2 mmol) and sodium hydroxide (9.36 g, 234.2 mmol) were added to 400 mL of tetrahydrofuran, followed by the slow dropwise addition of carbon disulfide (12.36 g, 281.4 mmol). After the addition was complete, the reaction was allowed to proceed at room temperature for 3 h. Upon completion of the reaction, a solid precipitated, which was filtered. The filter cake was washed 3-4 times with a small amount of tetrahydrofuran, and the dried filter cake yielded a brown solid, with a yield of 66.1%. MS (ESI) m / z: 520.1 [MH] - .

[0187] Preparation of step J 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate (Example 9)

[0188] At room temperature, intermediate A7 (1 g, 1.84 mmol) and potassium carbonate (0.38 g, 3.761 mmol) were added to 20 mL of LDM, followed by 2-chloro-N,N-dimethylacetamide (0.33 g, 2.21 mmol). The mixture was heated to 50 °C and reacted for 8 h. After the reaction was complete, the reaction solution was poured into 50 mL of ice water, and a solid precipitated out. The precipitate was filtered, and the filter cake was washed with a small amount of saturated sodium carbonate aqueous solution. The filter cake was purified by column chromatography to obtain a white solid with a yield of 57.9%. MS (ESI) m / z: 630.2 [M+Na] + ; 1H NMR (600MHz, DMSO-d6) δ7.93(d,J=8.4Hz,1H),7.73(t,J=52.9Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.0H z,1H),4.33(s,2H),4.12(d,J=40.0Hz,2H),3.97(s,6H),3.83(s,4H),3.70(s,4H),3.10(s,3H),2.85(s,3H).

[0189] Following the method of Example 2, Examples 10-16 were prepared by reacting intermediate A7 with a small molecule amine substituted with a haloacyl chloride.

[0190] Example 10: 2-(diethylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 658.2 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ7.93(s,1H),7.72(t,J=52.8Hz,1H),7.42(t,J=8.1Hz,1H),6.96(d,J=9.8Hz,1H),4.35(s,4H),4.16(s,2H) ,3.97(s,7H),3.82(s,4H),3.70(s,4H),3.43(q,J=7.1Hz,2H),3.29(q,J=7.1Hz,2H),1.21(t,J=7.1Hz,3H),1.02(t,J=7.1Hz,3H).

[0191] Example 11: 2-morpholino-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 672.2 [M+Na] + :672.21.Found:672.04; 1H NMR (600MHz, DMSO-d6) δ7.93(d,J=8.4Hz,1H),7.72(t,J=52.9Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.35(s,3 H),4.15(s,2H),3.97(s,7H),3.82(d,J=7.7Hz,4H),3.70(s,4H),3.63(d,J=4.5Hz,2H),3.60–3.54(m,4H),3.47–3.44(t,2H).

[0192] Example 12 2-(4-methylpiperazin-1-yl)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate; MS (ESI) m / z: 663.3 [M+H] + ; 1 H NMR (600MHz, DMSO-d6) δ7.93(d,J=8.3Hz,1H),7.73(t,J=52.9Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.35(s,3H), 4.15(d,J=11.5Hz,1H),3.97(s,5H),3.83(s,4H),3.70(s,4H),3.56(s,2H),3.46(s,2H),2.37(s,2H),2.26(s,2H),2.19(s,3H).

[0193] Example 13 2-O-2-(pyrrolidone-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 634.2 [M+H] + ; 1H NMR(600MHz,DMSO-d6)δ7.93(d,J=8.3Hz,1H),7.73(t,J=52.9Hz,1H),7.42 (t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.37(s,2H),4.25(s,2H),4.14(s,1 H),3.98(d,J=8.7Hz,7H),3.83(d,J=4.6Hz,4H),3.70(s,4H),3.58(t,J=6.8 Hz,2H),3.30(t,J=6.9Hz,2H),1.91(p,J=6.8Hz,2H),1.79(p,J=6.8Hz,2H).

[0194] Example 14: 2-(4-(methanesulfonyl)piperazin-1-yl)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate; MS (ESI) m / z: 727.2 [M+H] + ; 1 H NMR (600MHz, DMSO-d6) δ7.93(d,J=8.4Hz,1H),7.73(t,J=52.9Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.39(s,4H ),4.16(s,2H),3.97(s,7H),3.82(d,J=3.7Hz,5H),3.70(s,7H),3.58(s,3H),3.21(s,2H),3.10(s,2H),2.90(d,J=10.7Hz,5H).

[0195] Example 15 2-(4-cyclopentylpiperazin-1-yl)-oxyethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate; MS (ESI) m / z: 717.3 [M+H] + ; 1H NMR (600MHz, DMSO-d6) δ7.93(d,J=8.4Hz,1H),7.73(t,J=52.9Hz,1H),7.42(t,J=8.2 Hz,1H),6.96(d,J=8.1Hz,1H),4.37(d,J=24.7Hz,4H),4.15(s,2H),3.97(s,7H),3.82 (d,J=4.5Hz,4H),3.70(s,4H),3.55(d,J=0.5Hz,2H),3.44(d,J=0.4Hz,1H),2.46(s, 2H), 2.36 (s, 2H), 1.77 (s, 2H), 1.60 (d, J = 5.1Hz, 2H), 1.53–1.44 (m, 2H), 1.33 (s, 2H).

[0196] Example 16: 2-(cyclobutanoamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 656.2 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ8.43(d,J=7.6Hz,1H),7.93(d,J=8.2Hz,1H),7.72(t,J=52.9Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.0Hz,1H),4.37(s,2H) ,4.16(dd,J=16.4,7.8Hz,3H),4.02(s,2H),3.97(s,7H),3.83(s,4H),3.70 (s,4H),2.14(td,J=7.8,4.0Hz,2H),1.92–1.85(m,2H),1.69–1.55(m,2H).

[0197] Following the methods of Examples 1 and 2, a new intermediate was prepared from (S)-pyrrolidine-3-ylcarbamate / piperidine-4-ylcarbamate raw materials, and then reacted with a small molecule amine substituted with a haloacyl chloride to prepare Examples 17-18.

[0198] Example 17 2-O-2-(piperidin-1-yl)ethyl(S)-{1-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}pyrrolidine-3-yl}aminodithiocarbamate; MS (ESI) m / z: 648.2 [M+H] + .

[0199] Example 18: 2-O-2-(piperidin-1-yl)ethyl{1-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperidin-4-yl}aminodithiocarbamate; MS (ESI) m / z: 662.2 [M+H] + .

[0200] Following the method of Example 2, Examples 19-26 were prepared by reacting intermediate A7 as a raw material with a small molecule amine substituted with a haloalkyl group.

[0201] Example 19 2-Methoxyethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 581.2 [M+H] + ; 1 H NMR (600MHz, DMSO-d6) δ7.92(d,J=8.4Hz,1H),7.72(t,J=52.9Hz,1H),7.41(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.39(d,J=23. 7Hz, 2H), 4.11 (d, J = 21.2Hz, 2H), 3.97 (s, 7H), 3.82 (s, 4H), 3.70 (s, 4H), 3.57 (t, J = 6.4Hz, 2H), 3.51 (t, J = 6.0Hz, 2H), 3.27 (s, 3H).

[0202] Example 20: 2-(dimethylaminoethyl)4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 594.2 [M+H] + ; 1 H NMR (600MHz, DMSO-d6) δ7.92(d,J=8.4Hz,1H),7.72(t,J=52.9Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.43–4.3 4(t,2H),4.16–4.05(t,2H),3.97(s,7H),3.82(s,4H),3.70(s,4H),3.42(t,J=7.0Hz,2H),2.55(t,J=7.0Hz,2H),2.20(s,6H).

[0203] Example 21 2-(4-methylpiperazin-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate; MS (ESI) m / z: 649.3; 1 HNMR (600MHz, DMSO-d6) δ7.92(d,J=8.4Hz,1H),7.72(t,J=52.9Hz,1H),7.41(t,J=8.2Hz,1H),6.96(d,J=8.0Hz,1H),4.38(d,J=14.0H z,2H),4.10(d,J=13.5Hz,2H),3.97(s,7H),3.81(s,5H),3.70(s,4H),3.43(t,J=7.2Hz,2H),2.59(t,J=7.2Hz,2H),2.48–2.32(m,5H).

[0204] Example 22: 2-Morpholinyl ethyl ester 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 636.3 [M+H] + ; 1 H NMR (600MHz, DMSO-d6) δ7.93(d,J=8.4Hz,1H),7.72(t,J=52.9Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.38(t,J=5.7Hz,2H),4 .10(d,J=11.9Hz,2H),3.97(s,7H),3.82(s,4H),3.70(s,4H),3.58–3.56(t,4H),3.45(t,J=7.1Hz,2H),2.59(t,J=7.1Hz,2H),2.43(s,4H).

[0205] Example 23: 2-(piperidin-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 634.3 [M+H] + ; 1H NMR (600MHz, DMSO-d6) δ7.93(d,J=8.4Hz,1H),7.68(d,J=52.8Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.49–4.30(m, 2H),4.20–4.01(m,2H),3.97(s,7H),3.82(s,4H),3.70(s,4H),3.50–3.36(m,2H),2.51(s,2H),2.50(d,J=1.8Hz,4H),1.50(m,6H).

[0206] Example 24: 2-(pyrrolidone-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 620.2 [M+H] + ; 1 HNMR (600MHz, DMSO-d6) δ7.92(d,J=8.3Hz,1H),7.72(t,J=52.9Hz,1H),7.41(t,J=8.2Hz,1H),6.96(d,J=8.0Hz,1H),4.39(d,J=15.1 Hz,2H),4.10(d,J=17.2Hz,2H),3.97(s,7H),3.82(s,4H),3.70(s,4H),3.45(t,J=7.0Hz,2H),2.75(s,2H),2.54(s,3H),1.70(s,4H).

[0207] Example 25: Pyridin-4-ylmethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 614.2 [M+H] + .

[0208] Example 26: Cyanomethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 562.2 [M+H] + .

[0209] Examples 27-31 can be prepared by using intermediate A3 and tert-butyl 1,4-diaza-1-carboxylate / (S)-3-aminopyrrolidine-1-carboxylate / 4-aminopiperidine-1-carboxylate as raw materials and following steps E, F, G, and H of Example 1.

[0210] Example 27 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}-1,4-diaza-1-yl}2-oxoethylmorpholino-4-dithiocarbamate; MS (ESI) m / z: 664.2 [M+H] + .

[0211] Example 28: (S)-2-{3-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}amino}pyrrolidine-1-yl}-2-oxoethylmorpholino-4-dithiocarbamate; MS (ESI) m / z: 672.2 [M+Na] + ; 1 H NMR(600MHz, DMSO-d6)δ8.19–8.09(m,1H),7.86(t,J=54.5Hz,1H),7.46–7.34(m,1H),6.99–6.88(m,1H),4.61– 4.45(m,1H),4.34–4.12(m,4H),4.05–3.74(m,11H),3.74–3.55(m,10H),3.43–3.37(m,1H),2.35–1.96(m,2H).

[0212] Example 29 (S)-2-{3-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}amino}pyrrolidine-1-yl}-2-oxoethyl 4-(2-hydroxyethyl)piperazine-1-carbodisulfate; MS (ESI) m / z: 715.2 [M+Na] + 691.2 [MH] - ; 1H NMR (600MHz, DMSO-d6) δ8.20–8.09(m,1H),8.01–7.81(m,1H),7.44–7.34(m,1H),6.95(t,J=6.9Hz,1H),4.66–4.50(m,1H),4.47(s,1H),4.29–4. 15(m,4H),4.04–3.89(m,6H),3.84–3.66(m,10H),3.57–3.47(m,3H),3.4 0–3.35(m,1H),2.70–2.51(m,4H),2.45–2.38(m,2H),2.29–1.95(m,2H).

[0213] Example 30 27 2-{4-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}amino}piperidin-1-yl}-2-oxoethylmorpholino-4-dithiocarbamate; MS (ESI) m / z: 686.4 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ8.11(d,J=8.4Hz,1H),7.98–7.85(m,1H),7.47–7.35(m,1H),6.95(t,J=8.7Hz,1H),4.50–4.23(m,4H ),4.21(s,2H),4.09–4.02(m,2H),3.97(s,3H),3.95–3.60(m,14H),2.94–2.75(m,1H),2.05–1.88(m,2H),1.62–1.35(m,2H).

[0214] Example 31 2-{4-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}amino}piperidin-1-yl}-2-oxoethyl 4-(2-hydroxyethyl)piperazine-1-carbodisulfate; MS (ESI) m / z: 707.4 [M+H] + 729.4 [M+Na] + ; 1H NMR(600MHz,DMSO-d6)δ8.11(d,J=8.4Hz,1H),7.97–7.81(m,1H),7.47–7.34(m,1H) ,6.95(t,J=8.6Hz,1H),4.48(s,1H),4.44–4.24(m,4H),4.21(s,2H),4.13–4.03(m,2 H),3.97(s,3H),3.86–3.75(m,5H),3.73–3.64(m,4H),3.55–3.49(m,3H),2.92–2.75 (m,1H),2.68–2.54(m,2H),2.48–2.24(m,4H),2.08–1.86(m,2H),1.60–1.31(m,2H).

[0215] Examples 32-33 can be prepared by using o-phenylenediamine as a raw material and following steps B, C, D, E, F, G, and H of Example 1.

[0216] Example 32: 2-{4-{4-[2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylmorpholino-4-carbodisulfate; MS (ESI) m / z: 620.2 [M+H] + 642.1[M+Na] + ; 1 H NMR(600MHz,DMSO-d6)δ8.36(d,J=6.4Hz,1H),7.86(d,J=8.1Hz,1H),7.71(t,J=52.2Hz,1H),7.55–7.40(m,2H) ,4.41(s,2H),4.29–4.12(m,2H),4.07–3.91(m,4H),3.87–3.79(m,6H),3.78–3.66(m,10H),3.65–3.56(m,2H).

[0217] Example 33: 2-{4-{4-[2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl-4-ethylpiperazin-1-carbodisulfate; MS (ESI) m / z: 647.2 [M+H] + 669.2 [M+Na] + ; 1H NMR (600MHz, DMSO-d6) δ8.36(d,J=7.3Hz,1H),7.86(d,J=8.0Hz,1H),7.71(t,J=51.9Hz,1H),7.54–7.39(m,2H),4.39(s,2H),4.28–4.13(m,2H ),4.02–3.91(m,4H),3.86–3.79(m,6H),3.77–3.67(m,6H),3.65–3.57( m,2H),2.48–2.41(m,4H),2.37(q,J=7.0Hz,2H),1.02(t,J=7.1Hz,3H).

[0218] Preparation of Example 34:

[0219] Preparation of step K 1-(4,6-dichloropyrimidin-2-yl)-2-(difluoromethyl)-4-methoxy1H-benzo[d]imidazole (A8)

[0220] At room temperature, intermediate A3 (50.0 g, 252.3 mmol) and potassium carbonate (139.2 g, 100.9 mmol) were added to 500 mL of DMSO, followed by dropwise addition of 2,4,6-trichloropyrimidine (46.27 g, 252.3 mmol). The reaction mixture was allowed to react at room temperature for 6 h. After the reaction was complete, the reaction solution was poured into 1500 mL of water and stirred for 0.5 h. A solid precipitated, which was filtered. The filter cake was washed with a small amount of water and dried to obtain a gray solid with a yield of 78.9%. MS (ESI) m / z: 345.0 [M+H] + .

[0221] Preparation of step L 4-{6-chloro-2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]pyrimidin-4-yl}morpholine (A9)

[0222] At room temperature, intermediate A8 (50.0 g, 145.5 mmol) was added to 500 mL of THF, followed by dropwise addition of morpholine (12.66 g, 145.5 mmol). After the addition was complete, the reaction mixture was allowed to react at room temperature for 6 h. After the reaction was complete, the reaction solution was poured into 1500 mL of water and stirred for 0.5 h. A solid precipitated out; this was filtered, and the filter cake was washed with a small amount of water. After drying, a gray solid was obtained, with a yield of 81.2%. MS (ESI) m / z: 396.1 [M+H] + .

[0223] Following the steps of Example 1, using intermediate A9 as raw material, and preparing it via methods E, F, G, and H, Example 31, which yields a pyrimidine structure, can be obtained.

[0224] Example 34: 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethyl diethyl dithiocarbamate; MS (ESI) m / z: 635.2 [M+H] + ; 1 HNMR(600MHz, DMSO-d6δ7.78(d,J=8.3Hz,1H),7.65(t,J=53.0Hz,1H),7.36(t,J=8.2Hz,1H),6.90(d,J=8. 0Hz,1H),6.00(s,1H),3.96(s,3H),3.76(s,2H),3.73–3.65(m,8H),3.64–3.57(m,12H),1.74–0.66(m,5H).

[0225] The preparations of Examples 35-49 can be obtained by simply replacing the halogenated small molecule amine with the method of Example 31.

[0226] Example 35 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylpyrrolidine-1-carbodisulfate; MS (ESI) m / z: 655.0 [M+Na] + ; 1 HNMR (600MHz, DMSO-d6) δ7.79(d,J=8.3Hz,1H),7.66(t,J=53.0Hz,1H),7.38(t,J=8.2Hz,1H),6.91(d,J=8.0Hz,1H),6.01(s, 1H),4.41(s,2H),3.97(s,3H),3.83–3.77(m,2H),3.75–3.70(m,6H),3.70–3.66(m,6H),3.65–3.57(m,6H),1.44–0.76(m,4H).

[0227] Example 36: 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-4-carbodisulfate; MS (ESI) m / z: 664.9 [M+H] + 686.9 [M+Na] + ; 1H NMR(600MHz,DMSO-d6)δ7.79(d,J=8.3Hz,1H),7.66(t,J=53.0Hz,1H),7.38( t,J=8.2Hz,1H),6.91(d,J=8.0Hz,1H),6.01(s,1H),4.58–4.45(m,2H),4.40( s,2H),4.35–4.23(m,2H),3.97(s,3H),3.83–3.78(m,2H),3.75–3.69(m,6H) ,3.69–3.66(m,2H),3.65–3.62(m,4H),3.61–3.58(m,2H),2.78–2.68(m,4H).

[0228] Example 37 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylpiperidine-1-carbodisulfate; MS (ESI) m / z: 647.0 [M+H] + 668.9 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ7.79(d,J=8.3Hz,1H),7.66(t,J=53.0Hz,1H),7.38(t,J= 8.2Hz,1H),6.91(d,J=7.9Hz,1H),6.01(s,1H),4.37(s,2H),4.25–4.15(m,2H),3. 97(s,3H),3.96–3.86(m,2H),3.83–3.77(m,2H),3.75–3.69(m,6H),3.69–3.65(m ,2H),3.65–3.62(m,4H),3.61–3.58(m,2H),1.69–1.64(m,2H),1.63–1.54(m,4H);

[0229] Example 38 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethyl 4-(methanesulfonyl)piperazin-1-carbodisulfate; MS (ESI) m / z: 747.8 [M+Na] + ; 1H NMR (600MHz, DMSO-d6) δ7.79(d,J=8.3Hz,1H),7.66(t,J=53.0Hz,1H),7.38(t,J=8.2Hz,1H),6.91(d,1H),6.01(s,1H),4.41(s,2H),4.38–4.03 (m,4H),3.97(s,3H),3.84–3.77(m,2H),3.75–3.69(m,6H),3.69–3.66( m,2H),3.65–3.62(m,4H),3.62–3.58(m,2H),3.27(t,4H),2.94(s,3H).

[0230] Example 39 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-1,1-dioxide-4-carbodisulfate; MS (ESI) m / z: 697.2 [M+H] + 719.2 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ7.79(d,J=8.3Hz,1H),7.66(t,J=53.0Hz,1H),7.38(t,J=8.2Hz,1H),6.91(d,J=8.0Hz,1H ),6.01(s,1H),4.52(s,4H),4.41(s,2H),3.97(s,3H),3.94–3.76(m,3H),3.75–3.69(m,7H),3.69–3.52(m,10H).

[0231] Example 40: 2-(dimethylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 607.2 [M+H] + ; 1H NMR (600MHz, DMSO-d6) δ7.81(d,J=8.3Hz,1H),7.68(t,J=53.0Hz,1H),7.38(t,J=8.2Hz,1H),6.91(d,J=8.0Hz,1H),5.96(s,1H),4.3 5(d,J=25.3Hz,4H),4.13(s,2H),3.97(s,3H),3.86(d,J=8.7Hz,4H),3.74–3.68(m,4H),3.66–3.58(m,4H),3.10(s,3H),2.85(s,3H).

[0232] Example 41: 2-(diethylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 634.2 [M+H] + ; 1 HNMR(600MHz,DMSO-d6)δ7.81(d,J=8.3Hz,1H),7.68(t,J=53.0Hz,1H),7.38(t,J= 8.2Hz,1H),6.92(d,J=8.0Hz,1H),5.96(s,1H),4.36(d,J=13.1Hz,4H),4.13(s,2H) ,3.97(s,3H),3.86(d,J=11.7Hz,4H),3.75–3.68(t,4H),3.67–3.60(t,4H),3.43(q ,J=7.1Hz,2H),3.29(q,J=7.1Hz,2H),1.21(t,J=7.1Hz,4H),1.02(t,J=7.1Hz,3H).

[0233] Example 42: 2-O-2-(pyrrolidone-1-yl)ethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 654.8 [M+Na] + ; 1H NMR (600MHz, DMSO-d6) δ7.81(d,J=8.3Hz,1H),7.68(t,J=53.0Hz,1H),7.38(t,J=8.2H z,1H),6.91(d,J=8.0Hz,1H),5.96(s,1H),4.36(d,J=8.5Hz,2H),4.25(s,2H),4.13(s ,2H),3.97(s,3H),3.86(d,J=11.5Hz,4H),3.74–3.68(t,4H),3.67–3.61(t,4H),3.58 (t,J=6.8Hz,2H),3.30(t,J=6.9Hz,2H),1.91(p,J=6.8Hz,2H),1.78(p,J=6.8Hz,2H).

[0234] Example 43: 2-morpholino-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 648.2 [M+H] + ; 1 H NMR (600MHz, DMSO-d6) δ7.81(d,J=8.2Hz,1H),7.68(t,J=53.0Hz,1H),7.38(t,J=8.2Hz,1H),6.92(d,J=7.9Hz,1H),5.96(s,1H),4.35 (s,4H),4.13(s,2H),3.97(s,3H),3.86(d,J=14.9Hz,4H),3.73–3.69(t,4H),3.66–3.60(t,6H),3.60–3.53(m,4H),3.48–3.42(t,2H).

[0235] Example 44: 2-(cyclobutanylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 633.2 [M+H] + ; 1H NMR (600MHz, DMSO-d6) δ8.43(d,J=7.6Hz,1H),7.81(d,J=8.3Hz,1H),7.68(t,J=53.1Hz,1H), 7.38(t,J=8.2Hz,1H),6.91(d,J=8.0Hz,1H),5.96(s,1H),4.36(s,2H),4.17(dd,J=16.3,8.0 Hz,2H),4.11(s,1H),4.02(s,2H),3.97(s,3H),3.85(d,J=7.4Hz,4H),3.73–3.69(t,4H),3.6 4(d,J=4.7Hz,4H),2.14(dtd,J=10.3,7.9,2.5Hz,2H),1.96–1.84(m,2H),1.69–1.57(m,2H).

[0236] Example 45: 2-Methoxyethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 580.2 [M+H] + ; 1 H NMR (600MHz, DMSO-d6) δ7.80(d,J=8.3Hz,1H),7.68(t,J=53.1Hz,1H),7.38(t,J=8.2Hz,1H),6.91(d,J=8.0Hz,1H),5.96(s,1H),4.39(d,J=4. 6Hz,2H),4.09(d,J=6.3Hz,2H),3.97(s,4H),3.85(s,4H),3.67(dt,J=9 .3,4.4Hz,9H),3.54(dt,J=34.4,6.1Hz,3H),3.32(s,3H),3.27(s,2H).

[0237] Example 46: 2-(dimethylamino)4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 593.2 [M+H] + ; 1H NMR (600MHz, DMSO-d6) δ7.80(d,J=8.2Hz,1H),7.68(t,J=53.0Hz,1H),7.38(t,J=8.2Hz,1H),6.91(d,J=8.0Hz,1H),5.95(s,1H),4.38(s,2 H), 4.09 (s, 2H), 3.97 (s, 3H), 3.84 (d, J = 4.6Hz, 4H), 3.67 (dt, J = 9.4, 4.4Hz, 8H), 3.42 (t, J = 7.0Hz, 2H), 2.56 (t, J = 6.9Hz, 2H), 2.21 (s, 6H).

[0238] Example 47: 2-(pyrrolidone-1-yl)ethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 619.2 [M+H] + .

[0239] Example 48: Pyridin-4-ylmethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 613.2 [M+H] + .

[0240] Example 49: Cyanomethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 561.2 [M+H] + .

[0241] Example 50: 2-Amino-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 614.2 [M+H] + ; 1H NMR (600MHz, DMSO-d6) δ7.93(d,J=8.3Hz,1H),7.73(t,J=52.9Hz,1H),7.58(s,1H),7.42(t,J=8.2Hz,1H),7.19(s,1H),6.96 (d,J=8.1Hz,1H),4.38(dd,J=12.5,6.8Hz,2H),4.13(d,J=26.3Hz,2H),4.04(s,2H),3.97(s,6H),3.83(s,4H),3.70(s,4H).

[0242] Example 51: 2-(cyclohexylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 684.2 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ8.05(d,J=7.8Hz,1H),7.93(d,J=8.3Hz,1H),7.72(t,J=52.9Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.36(s, 2H),4.14(s,1H),4.02(s,2H),3.97(s,5H),3.82(d,J=3.4Hz,4H),3.70(s ,4H),3.52(dd,J=16.2,8.5Hz,1H),1.81–1.62(m,4H),1.33–1.06(m,6H).

[0243] Example 52: 2-(cyclopentylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 669.2 [M+Na] + ; 1H NMR(600MHz,DMSO-d6)δ8.15(d,J=7.2Hz,1H),7.93(d,J=8.4Hz,1H),7.72(t, J=52.9Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.37(s,2H),4.12 (s,2H),4.01(s,2H),3.98(d,J=6.7Hz,8H),3.82(d,J=4.7Hz,4H),3.70(s,4H ),1.82–1.73(m,2H),1.68–1.58(m,2H),1.53–1.46(m,2H),1.43–1.35(m,2H).

[0244] Example 53: 2-(cyclopropylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 642.2 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ8.28(d,J=3.1Hz,1H),7.93(d,J=8.4Hz,1H),7.73(t,J=52.9Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.48–4.2 8(t,2H),4.18–4.07(t,2H),4.00(s,3H),3.97(s,6H),3.83(s,4H),3.70( s,5H),2.62(tq,J=7.6,3.9Hz,1H),0.68–0.56(m,2H),0.46–0.36(m,2H).

[0245] Example 54: 2-(cyclopropylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 619.2 [M+H] + ; 1HNMR (600MHz, DMSO-d6) δ8.24(d,J=3.8Hz,1H),7.81(d,J=8.3Hz,1H),7.68(t,J=53. 0Hz,1H),7.38(t,J=8.2Hz,1H),6.91(d,J=8.0Hz,1H),5.96(s,1H),4.35(d,J=8.0Hz, 2H),4.10(d,J=11.0Hz,2H),3.98(d,J=16.4Hz,5H),3.86(d,J=10.0Hz,4H),3.67(dt, J=9.1,4.3Hz,9H),2.62(tq,J=7.7,3.9Hz,1H),0.70–0.55(m,2H),0.49–0.31(m,2H).

[0246] Example 55: 2-Amino-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; MS (ESI) m / z: 601.2 [M+H] + ; 1 H NMR (600MHz, DMSO-d6) δ7.81(d,J=8.3Hz,1H),7.68(t,J=53.1Hz,1H),7.58(s,1H),7.38(t,J=8.2Hz,1H),7.19(s,1H),6.92(d,J=7.9Hz,1H ),5.96(s,1H),4.38(d,J=5.9Hz,2H),4.12(d,J=1.3Hz,2H),4.04(s,2H),3.97(s,3H),3.86(d,J=13.5Hz,4H),3.68(dt,J=9.3,4.4Hz,8H).

[0247] Example 56: 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-4-dithiocarbamate; MS (ESI) m / z: 665.8 [M+H] + 687.7 [M+Na] + ; 1H NMR (600MHz, DMSO-d6) δ7.91(d,J=7.7Hz,1H),7.71(t,J=52.9Hz,1H),7.41(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4.64–4.43(m,2H),4. 41(s,2H),4.38–4.19(m,2H),3.97(s,3H),3.95–3.89(m,2H),3.87–3 .78(m,6H),3.76–3.67(m,6H),3.64–3.58(m,2H),2.80–2.67(m,4H).

[0248] Example 57: 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylcyclopropylaminodithiocarbamate; MS (ESI) m / z: 619.8 [M+H] + 641.8 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ7.90(d,J=5.3Hz,1H),7.71(t,J=52.7Hz,1H),7.42(t,J=8.2Hz,1H),6.96(d,J=8.1Hz,1H),4. 38–4.28(m,1H),3.97(s,3H),3.93(s,2H),3.90–3.76(m,8H),3.75–3.56(m,8H),1.31–1.18(m,2H),0.85–0.65(m,2H).

[0249] Example 58: 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 3,5-dimethylpiperazin-1-dithiocarbamate; MS (ESI) m / z: 676.8 [M+H] + ; 1H NMR (600MHz, DMSO-d6) δ7.91(d,J=7.8Hz,1H),7.71(t,J=52.7Hz,1H),7.42(t,J=8.2Hz ,1H),6.96(d,J=8.1Hz,1H),5.30–5.17(m,1H),4.48–4.40(m,1H),4.39(s,2H),3.97(s ,3H),3.96–3.89(m,2H),3.88–3.77(m,6H),3.76–3.65(m,6H),3.65–3.53(m,2H),2.90 –2.81(m,1H),2.77–2.68(m,2H),2.65–2.57(m,1H),1.23(s,1H),1.03(d,J=7.4Hz,6H).

[0250] Example 59 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-4-dithiocarboxylic acid 1,1-dioxide; MS (ESI) m / z: 719.9 [M+Na] + ; 1 H NMR (600MHz, DMSO-d6) δ7.91(d,1H),7.71(t,J=52.4Hz,1H),7.42(t,J=7.9Hz,1H),6.96(d,J=7.6Hz,1H),5.82–5. 68(m,2H),4.89–4.42(m,4H),4.41(s,2H),3.97(s,3H),3.96–3.91(m,2H),3.89–3.65(m,14H),3.65–3.51(m,2H);

[0251] Example 60: 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethyl-4-cyclopentylpiperazin-1-dithiocarbamate; MS (ESI) m / z: 716.0 [M+H] + ; 1H NMR (600MHz, DMSO-d6) δ7.79(d,J=8.3Hz,1H),7.66(t,J=53.0Hz,1H),7.38(t,J=8.2Hz,1H),6.9 1(d,1H),6.01(s,1H),4.38(s,2H),4.26–4.13(m,2H),3.97(s,3H),3.97–3.86(m,2H),3.83–3.7 7(m,2H),3.75–3.69(m,6H),3.67(s,2H),3.65–3.62(m,4H),3.62–3.58(m,2H),2.55–2.51(m,1H ),2.49–2.44(m,4H),1.81–1.73(m,2H),1.65–1.56(m,2H),1.53–1.44(m,2H),1.40–1.29(m,2H).

[0252] Study on the antitumor activity and PI3K inhibitory activity of the compounds of this invention

[0253] PI3Kα kinase activity assay

[0254] The ADP-Glo ​​luminescent kinase assay is performed through the following steps:

[0255] ① Prepare compound solutions: Dilute the test compounds (the compounds prepared in the above examples) in DMSO to a final concentration range of 0.08 nM-50 nM. Add 50 nL of each solubility solution of each test compound to a 384-well plate.

[0256] ② Prepare the kinase solution: Prepare a PI3Kα solution using 1× kinase buffer, at twice the final concentration. Add 2.5 μL of kinase solution to the test wells and 2.5 μL of 1× kinase buffer to the control wells.

[0257] ③ Prepare substrate solutions: Prepare solutions of ATP and PIP2 using 1× kinase buffer at twice the final concentration. Add 2.5 μL of the substrate solution of ATP and PIP2 to each well.

[0258] ④ Kinase reaction: Cover the test plate and incubate at room temperature for 1 hour;

[0259] ⑤ Kinase assay: Add 5 μL of ADP-Glo ​​reagent to each well and incubate for another 2 h. Then add 10 μL of kinase assay reagent to each well and shake the plate for 1 min.

[0260] ⑥ Read the plate and calculate the results: Use a microplate reader to read the RLU values ​​from the plate, and use normalized dose-response fitting nonlinear regression to calculate the IC50 using XLFit 5.4.0.8 software.50 The value, specific steps, and formula are as follows:

[0261] (I) Use a microplate reader to read the RLU value of each well; (II) Percent inhibition = (RLU) max -RLU sample ) / (RLU max -RLU min )×100, where RLU max DMSO control group, RLU min Enzyme-free control group, RLU sample (III) Enter the drug concentration and inhibition rate into XLFit 5.4.0.8 and fit the data to obtain the IC50. 50 value.

[0262] Table 1 shows the inhibition data of some of the compounds in the examples on PI3Kα kinase.

[0263] Table 1

[0264]

[0265] As shown in Table 1 above, all compounds described in this invention exhibit inhibitory activity against PI3Kα kinase, with some compounds showing particularly pronounced effects. Further experiments were then conducted using the compounds described in this invention to investigate their in vitro antitumor cell activity.

[0266] The 1,3,5-triazine / pyrimidine compounds containing benzimidazole structures according to Formula I of the present invention were screened in vitro for their inhibitory activity against human colon cancer cells HCT-116, human bladder cancer cells PC-3, and human breast cancer cells MCF-7.

[0267] (1) After reviving and stabilizing the cells using standard methods by passage 2-3 times, the cells were digested from the bottom of the culture flask using trypsin solution (0.25%). The digested cells were poured into centrifuge tubes, followed by the addition of culture medium to terminate the digestion. The centrifuge tubes were centrifuged at 800 rpm for 10 min. The supernatant was discarded, and 5 mL of culture medium (RPMI-1640 medium with 10% fetal bovine serum and 1% penicillin antibiotics) was added. The cells were mixed by pipetting, and 10 μL of the cell suspension was added to a cell counting chamber for counting. The cell concentration was adjusted to 10-1. 4 Cells / well. Except for well A1, which was a blank well with no cells, 100 μL of cell suspension was added to all other wells of the 96-well plate. The 96-well plate was then incubated in an incubator for 24 hours.

[0268] (2) Dissolve the compound prepared in the above example in 50 μL of dimethyl sulfoxide as the test sample, and then add an appropriate amount of culture medium (RPMI-1640 medium with 10% fetal bovine serum and 1% penicillin in this example) to dissolve the sample into a 2 mg / mL solution. Then dilute the sample in a 24-well plate to 20, 4, 0.8, 0.16 and 0.032 μg / mL.

[0269] Each concentration was added to 3 wells, with the outer two rows and two columns of cells, whose growth was more significantly affected by the environment, used only as a blank control group. The 96-well plates were placed in an incubator and incubated for 72 hours.

[0270] (3) Discard the drug-containing culture medium in the 96-well plate, wash the cells twice with phosphate-buffered saline (PBS), add 100 μL of MTT (0.5 mg / mL) to each well, incubate for 4 hours, then discard the MTT solution and add 100 μL of dimethyl sulfoxide. Shake on a magnetic oscillator to fully dissolve the surviving cells and the MTT reaction product formazan, then measure the results using a microplate reader. The IC50 of the drug can be determined using the Bliss method. 50 value.

[0271] The results of the compounds' inhibitory activity against human colon cancer cells HCT-116, human bladder cancer cells PC-3, and human breast cancer cells MCF-7 are shown in Table 2.

[0272] Table 2

[0273]

[0274] As can be seen from the above efficacy data, the compound represented by general formula I of the present invention can interact with the PI3K protein due to the benzimidazole-1,3,5-triazine / pyrimidine structure, thus exhibiting good antitumor activity and significant inhibitory activity against PI3K. At the same time, it has a significant inhibitory effect on a variety of human tumor cells, especially human colon cancer cells HCT-116 and human bladder cancer cells PC-3.

Claims

1. A benzimidazole derivative, characterized in that: Derivatives are compounds represented by general formula I and their pharmaceutically acceptable salts. in, X represents N and CH; R1 is hydrogen, an unsubstituted (C1-C6) alkoxy group, or a (C3-C6) cycloalkoxy group substituted with at least one of the following groups: hydroxyl, amino, or dimethylamino. Ring A is selected from , , ; L is , ; Y represents C=O and CH2; When L is L1, R2 is -NR3R4; when L is L2, R2 is -NR3R4, hydroxyl, amino, carboxyl, (C1–C6)alkoxy, (C1–C6)alkoxyformyl, (C1–C6)alkylsulfonyl. R3 and R4 may be the same or different, and are independently selected from hydrogen, (C2-C6) alkenyl, (C2-C6) alkynyl, unsubstituted or substituted with 1-3 identical or different R6, (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl, and dimethylaminoethyl; Alternatively, R3 and R4 together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclic group containing at least one heteroatom; the heterocyclic group optionally contains 1-4 heteroatoms selected from N, O and / or S, the heterocyclic group optionally is substituted by one or more oxo atoms, and the heterocyclic group optionally is substituted by 0-3 identical or different R7 atoms. R6 and R7 may be the same or different, and are independently selected from hydrogen, hydroxyl, halogen, amino group substituted with 0-2 (C1-C6) alkyl groups, cyano group, (C1-C6) alkyl acyl group, (C1-C6) alkyl sulfonyl group, unsubstituted (C1-C6) alkyl group or substituted with at least one of the following groups, (C1-C6) alkoxy group, (C3-C6) cycloalkyl group; the following groups are hydroxyl, amino or halogen; n is an integer between 1 and 2; m is 1.

2. The benzimidazole derivative according to claim 1, characterized in that: The derivative is a compound of general formula I and its pharmaceutically acceptable salt, wherein... X represents N and CH; R1 is a hydrogen, an unsubstituted (C1-C4) alkoxy group, or a group substituted with at least one of the following groups: hydroxyl, amino, or dimethylamino. Ring A is selected from , , ; L is , ; Y represents C=O and CH2; When L is L1, R2 is -NR3R4; when L is L2, R2 is -NR3R4, hydroxyl, amino, carboxyl, (C1–C4)alkoxy, (C1–C4)alkoxyformyl, (C1–C4)alkylsulfonyl. R3 and R4 may be the same or different, and are independently selected from hydrogen, dimethylaminoethyl, unsubstituted or substituted with 1-3 identical or different R6 (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl; Alternatively, R3 and R4 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing at least one heteroatom; the heterocyclic group optionally contains 1-4 heteroatoms selected from N, O and / or S, the heterocyclic group optionally is substituted by one or more oxo atoms, and the heterocyclic group optionally is substituted by 0-3 identical or different R7 atoms. R6 and R7 may be the same or different, and are independently selected from hydrogen, hydroxyl, halogen, amino group substituted with 0-2 (C1-C4) alkyl groups, cyano group, (C1-C4) alkyl acyl group, (C1-C4) alkyl sulfonyl group, unsubstituted (C1-C4) alkyl group or substituted with at least one of the following groups, (C1-C4) alkoxy group, (C3-C6) cycloalkyl group; the following groups are hydroxyl, amino or halogen; n is an integer between 1 and 2; m is 1.

3. The benzimidazole derivative according to claim 2, characterized in that: The derivatives are compounds of general formula I and their pharmaceutically acceptable salts. X represents N and CH; R1 is hydrogen or (C1-C4)alkoxy; Ring A is selected from , , ; L is , ; Y represents C=O and CH2; When L is L1, R2 is -NR3R4; when L is L2, R2 is -NR3R4, hydroxyl, amino, carboxyl, (C1–C4)alkoxy, (C1–C4)alkoxyformyl, (C1–C4)alkylsulfonyl. R3 and R4 may be the same or different, and are independently selected from hydrogen, dimethylaminoethyl, unsubstituted or substituted with 1-3 identical or different R6 (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl; Or R3 and R4 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing at least one heteroatom; The heterocyclic group optionally contains 1-4 heteroatoms selected from N, O and / or S, the heterocyclic group optionally is substituted by one or more oxygen atoms, and the heterocyclic group optionally is substituted by 0-3 identical or different R7 atoms; R6 and R7 may be the same or different, and are independently selected from hydrogen, hydroxyl, halogen, amino group substituted with 0-2 (C1-C4) alkyl groups, cyano group, (C1-C4) alkyl acyl group, (C1-C4) alkyl sulfonyl group, unsubstituted (C1-C4) alkyl group or substituted with at least one of the following groups, (C1-C4) alkoxy group, (C3-C6) cycloalkyl group; the following groups are hydroxyl, amino or halogen; n is an integer between 1 and 2; m is 1.

4. The benzimidazole derivative according to claim 3, characterized in that: The derivative is a compound of general formula I and its pharmaceutically acceptable salt, wherein... X represents N and CH; R1 is hydrogen or methoxy; Ring A is selected from , , ; L is , ; Y represents C=O and CH2; When L is L1, R2 is -NR3R4; when L is L2, R2 is -NR3R4, hydroxyl, amino, carboxyl, (C1–C4)alkoxy, (C1–C4)alkoxyformyl, (C1–C4)alkylsulfonyl. R3 and R4 may be the same or different, and are independently selected from hydrogen, dimethylaminoethyl, unsubstituted or substituted with 1-3 identical or different R6 (C1-C6) alkyl, (C3-C7) cycloalkyl, (C3-C6) heterocycloalkyl; Or R3 and R4 together with the nitrogen atoms they are attached to form , , , , , , , , , , , , , , , , , , , , , , ; n is an integer between 1 and 2; m is 1.

5. The benzimidazole derivative according to claim 4, characterized in that: The derivatives are compounds of general formula I and their pharmaceutically acceptable salts. 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyldiethyldithiocarbamate; 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-yl}-2-oxoethylpyrrolidine-1-carbodisulfate; 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-yl}-2-oxoethylmorpholino-4-carbodisulfate; 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-yl}-2-oxoethylpiperidine-1-carbodisulfate; 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazin-1-yl}-2-oxoethyl 4-methylpiperazine-1-carbodisulfate; 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 4-(methanesulfonyl)piperazin-1-carbodisulfate; 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 4-(2-hydroxyethyl)piperazin-1-carbodisulfate; 2-{4-{4-[(2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazin-1-yl}-2-oxoethyl-4-ethylpiperazine-1-carbodisulfate; 2-(dimethylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-(diethylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-morpholino-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-(4-methylpiperazin-1-yl)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate; 2-Oxo-2-(pyrrolidone-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate; 2-(4-(methanesulfonyl)piperazin-1-yl)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate; 2-(4-Cyclopentylpiperazin-1-yl)-oxyethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate; 2-(cyclobutanylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-Oxo-2-(piperidin-1-yl)ethyl(S)-{1-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}pyrrolidine-3-yl}aminodithiocarbamate; 2-Oxo-2-(piperidin-1-yl)ethyl{1-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperidin-4-yl}aminodithiocarbamate; 2-Methoxyethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-(dimethylaminoethyl)4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-(4-methylpiperazin-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazin-1-dithiocarbamate; 2-Morpholinyl ethyl ester 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-(piperidin-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazin-2-yl}piperazine-1-dithiocarbamate; 2-(pyrrolidone-1-yl)ethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; Pyridin-4-ylmethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; cyanomethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}-1,4-diaza-1-yl}2-oxoethylmorpholino-4-dithiocarbamate; (S)-2-{3-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}amino}pyrrolidine-1-yl}-2-oxoethylmorpholino-4-dithiocarbamate; (S)-2-{3-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}amino}pyrrolidine-1-yl}-2-oxoethyl 4-(2-hydroxyethyl)piperazine-1-carbodisulfate; 2-{4-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}amino}piperidin-1-yl}-2-oxoethylmorpholino-4-dithiocarbamate; 2-{4-{{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazine-2-yl}amino}piperidin-1-yl}-2-oxoethyl 4-(2-hydroxyethyl)piperazine-1-carbodisulfate; 2-{4-{4-[2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylmorpholino-4-carbodisulfate; 2-{4-{4-[2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 4-ethylpiperazin-1-carbodisulfate; 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethyl diethyl dithiocarbamate; 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylpyrrolidine-1-carbodisulfate; 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-4-carbodisulfate; 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylpiperidine-1-carbodisulfate; 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethyl 4-(methanesulfonyl)piperazin-1-carbodisulfate; 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-1,1-dioxide-4-carbodisulfate; 2-(dimethylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; 2-(diethylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; 2-O-2-(pyrrolidone-1-yl)ethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; 2-morpholino-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; 2-(cyclobutanylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; 2-Methoxyethyl 4-{2-[2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; 2-(dimethylamino)4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; 2-(pyrrolidone-1-yl)ethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; Pyridin-4-ylmethyl 4-{2-[2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; cyanomethyl 4-{2-[2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; 2-Amino-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-(cyclohexylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-(cyclopentylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-(cyclopropylamino)-2-oxoethyl 4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholin-1,3,5-triazine-2-yl}piperazine-1-dithiocarbamate; 2-(cyclopropylamino)-2-oxoethyl 4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; 2-Amino-2-oxoethyl 4-{2-[2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazine-1-dithiocarbamate; 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-4-dithiocarbamate; 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylcyclopropylaminodithiocarbamate; 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethyl 3,5-dimethylpiperazin-1-dithiocarbamate; 2-{4-{4-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholino-1,3,5-triazin-2-yl}piperazin-1-yl}-2-oxoethylthiomorpholino-4-dithiocarboxylic acid 1,1-dioxide; 2-{4-{2-[2-(difluoromethyl)-4-methoxy-1H-benzo[d]imidazol-1-yl]-6-morpholinopyrimidin-4-yl}piperazin-1-yl}-2-oxoethyl-4-cyclopentylpiperazin-1-dithiocarbamate.

6. The benzimidazole derivative according to any one of claims 1-5, characterized in that: The compound salt is a salt formed with an acid 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 a compound of general formula I according to any one of claims 1-5 and a pharmaceutically acceptable salt thereof.

8. The use of a compound of general formula I according to claim 1 and a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound of general formula I according to claim 1 and a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating and / or preventing PI3K-mediated diseases.

9. The application according to claim 8, characterized in that: The diseases that are prevented from being PI3K mediated include lung cancer, liver cancer, stomach cancer, colorectal cancer, breast cancer, glioblastoma, bladder cancer, prostate cancer, ovarian cancer, or esophageal cancer.

Images