PRMT7 target inhibitor and application thereof

Abstract

The invention discloses a PRMT7 target inhibitor and application thereof, the general formula of the compound is shown as any one of formulas (II)-(V), the invention discloses a plurality of compounds with novel structures, and the compounds can inhibit PRMT7 so as to resist tumors, have the advantages of high activity, good treatment effect and the like, and can be used for preparing antitumor drugs. # imgabs0 # imgabs1 #

Description

[0001] The present invention is a divisional application with the invention name: A PRMT7 target inhibitor and its application, application number: 2024112753072, and application date: 2024.9.12. Technical Field

[0002] The present invention relates to a PRMT7 target inhibitor and application thereof, and belongs to the field of medicinal chemistry. Background Art

[0003] Protein arginine methylation has attracted much attention due to the cell biological processes it mediates and the physiological and disease processes it participates in. Protein arginine methyltransferases (PRMTs), the enzymatic reaction of this post-translational modification, have gradually become an important target that has attracted much attention in the field of drug discovery.

[0004] Nine sequence-related PRMT subtypes (PRMT1-9) have been identified in mammals, which are divided into three categories (type I, type II, and type III) based on the products of the methylation reaction. Type I PRMTs (PRMT1, PRMT2, PRMT3, PRMT4, PRMT6, and PRMT8) catalyze the formation of monomethylarginine (Rme1) and asymmetric dimethylarginine (Rme2a), type II PRMTs (PRMT5 and PRMT9) catalyze the formation of Rme1 and symmetric dimethylarginine (Rme2s), and type III PRMT (PRMT7) only catalyzes the formation of Rme1. The nine members of the PRMT protein family have a common Rossmann-like structure, including seven β-sheets connected by α-helices and a β-barrel domain. They and some non-SET domain lysine methyltransferases (such as DOT1L) are classified as class I S-adenosylmethionine (SAM)-dependent methyltransferases, which have the same basic domain structure. However, little is known about this methyltransferase. All PRMTs except PRMT7 and PRMT9 contain only one methyltransferase domain, which in turn contain two tandem domains that arose from an ancestral gene duplication. The C-terminal domain alone is catalytically inactive, but folds together with the N-terminal domain to form a pseudodimer that is essential for enzyme activity.

[0005] PRMT7 plays an important biological function in normal physiological processes, and its disorder is closely related to the occurrence and development of various cancers. In breast cancer, high expression of PRMT7 promotes the proliferation and metastasis of tumor cells, and is a potential target for breast cancer treatment. In chronic myeloid leukemia (CML), PRMT7 maintains the survival of cancer stem cells (CSC) by regulating glycine metabolism. Gene knockout of PRMT7 can effectively kill CML cells without affecting normal cells, making it a very promising therapeutic target for CML. So far, several PRMT inhibitors have been discovered, mainly type I PRMT inhibitors, including selective and non-selective inhibitors, as well as PRMT5 inhibitors. In 2020, Szewczyk et al. reported the first PRMT7 selective inhibitor SGC3027, which had an inhibitory activity (IC 50 ) reached 2.5 nM, but it is an adenosine analogue with poor drugability and severe toxic side effects. Recently, Yang et al. reported a new class of 5-fluoro-4-aminopyrimidine derivatives, which have activity against PRMT7 at only a few micromolar levels. The best compound JS1310 selectively inhibited PRMT7 in the PRMTs family, with an IC 50 Therefore, it is of great significance to design and synthesize novel PMRT7 inhibitors with high selectivity, high activity, low toxicity and good drugability through medicinal chemistry methods and means. Summary of the invention

[0006] Purpose of the invention: The technical problem to be solved by the present invention is to provide a PRMT7 target inhibitor having the advantages of high activity and good therapeutic effect and its application.

[0007] Technical solution: To solve the above technical problems, the present invention provides a compound or a pharmaceutically acceptable salt thereof for inhibiting the activity of PRMT7 protein, wherein the general formula of the compound is as shown in any one of formulas (I) to (V): ; Wherein, when X1 and X2 are independently selected from sulfur atoms, CH; R1 is selected from the following groups: ; Wherein, R2, R3, R4, R5, and R6 are independently selected from methoxy, ethyl, and hydrogen; A and B are independently selected from CH or N.

[0008] Wherein, when X1 is a sulfur atom, X2 is CH; when X1 is CH, X2 is a sulfur atom; R1 is selected from the following groups: .

[0009] The present invention also provides a compound for inhibiting the activity of PRMT7 protein or a pharmaceutically acceptable salt thereof, the structural formula of which is shown below: .

[0010] Wherein, the pharmaceutically acceptable salt is an acid addition salt of the compound.

[0011] Among them, the acid used for salt formation is an inorganic acid or an organic acid, the inorganic acid includes hydrochloric acid, sulfuric acid or phosphoric acid; the organic acid includes acetic acid, trichloroacetic acid, propionic acid, butyric acid, maleic acid, p-toluenesulfonic acid, malic acid, malonic acid, cinnamic acid, citric acid, fumaric acid, camphoric acid, digluconic acid, aspartic acid, tartaric acid or methanesulfonic acid.

[0012] The present invention also provides a pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt, enantiomer thereof, and a pharmaceutically acceptable carrier.

[0013] The present invention also provides use of the compound or a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of a PRMT7 target inhibitor drug.

[0014] The present invention also provides the use of the compound or its pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of a drug for preventing and / or treating cancer or tumor-related diseases.

[0015] Wherein, the cancer or tumor-related diseases include prostate cancer, colorectal cancer, ovarian cancer, leukemia, lung cancer, liver cancer, breast cancer, esophageal cancer, glioma or pancreatic cancer.

[0016] The present invention designs and synthesizes multiple novel PRMT7 inhibitors through multiple medicinal chemistry methods and means, and tests their inhibitory activity on PRMT7 and anti-proliferative activity on multiple tumor cell lines.

[0017] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: The present invention provides a variety of compounds with novel structures, which can inhibit PRMT7 and thus fight tumors, have the advantages of high activity and good therapeutic effect, and can be used to prepare anti-tumor drugs. DETAILED DESCRIPTION

[0018] The technical solution of the present invention is further described below.

[0019] Example 1: Preparation of 2-methyl-N 6 -(7-(3,4,5-trimethoxyphenyl)quinoxaline-2-yl)quinoline-4,6-diamine (I-1) Synthesis route:

[0020] Synthesis method: Step 1: 2-cyano-4-nitroaniline (6.53 g, 40 mmol), 30 mL acetone, and 90 mL toluene were placed in a 250 mL round-bottom flask, cooled to 0 °C in an ice bath, and then 6.0 mL SnCl4 was slowly added dropwise. After the addition was completed, the reaction system was slowly raised to room temperature, then heated to 110 °C and refluxed for 6 hours, filtered, washed three times with dichloromethane (20 mL × 3), washed three times with methanol (10 mL × 3), and dried to obtain a yellow solid A1. 1 H NMR (400 MHz, DMSO- d 6) δ 9.46 (d, J = 2.4 Hz, 1H), 9.21 (s, 2H), 8.61 (dd, J = 9.3, 2.4 Hz, 1H), 8.03 (d, J = 9.3 Hz,1H), 6.71 (s, 1H), 2.62 (s, 3H). Step 2: A1 (6.0 g, 29.53 mmol), 200 mL of water and 1.2 g of 10% Pd / C were placed in a 500 mL round-bottom flask, reacted with hydrogen at room temperature overnight, filtered, and the filtrate was concentrated under reduced pressure to obtain a yellow solid A2 (4.86 g, 95%). 1 H NMR (400MHz, DMSO- d 6) δ 8.38 (brs, 2H), 7.78 (d, J = 8.9 Hz, 1H), 7.29 (dd, J = 9.0, 2.3Hz, 1H), 7.21 (d, J = 2.3 Hz, 1H), 6.51 (s, 1H), 5.76 (s, 2H), 2.54 (s, 3H). Step 3, A2 (173 mg, 1.0 mmol), 7-bromo-2-chloroquinoxaline (292 mg, 1.2 mmol), tris(dibenzylideneacetone)dipalladium (Pd2dba3, 46 mg, 0.05 mmol), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (Xantphos, 58 mg, 0.1 mmol), sodium tert-butoxide (290 mg, 3 mmol) and 5 mL 1,4-dioxane were placed in a 25 mL round-bottom flask, argon was replaced three times, and the reaction was carried out at 50° C. under argon protection overnight. The reaction solution was filtered through diatomaceous earth, concentrated under reduced pressure, and purified by a rapid silica gel column to obtain a yellow solid A3 (61 mg, 16%). 1 H NMR (400 MHz, DMSO- d 6) δ10.71 (s, 1H), 9.03 (d, J = 2.2 Hz, 1H), 8.73 (s, 1H), 8.46 – 8.22 (m, 3H), 8.08 (dd, J = 9.1, 2.2 Hz, 1H), 7.91 (d, J = 9.0 Hz, 1H), 7.83 (d, J = 8.6 Hz, 1H),7.64 (dd, J = 8.7, 2.2 Hz, 1H), 6.64 (s, 1H), 2.58 (s, 3H). Step 4, take A3 (61 mg, 0.16 mmol), 3,4,5-trimethoxyphenylboronic acid (41 mg, 0.19 mmol), bistriphenylphosphine palladium dichloride (5.6 mg, 0.008 mmol, Titan reagent, product number 1020950), sodium carbonate (51 mg, 0.48 mmol), 3 mL 1,4-dioxane and 0.2 mL water in a 25 mL round-bottom flask, replace argon three times, and react at 80 ° C under argon protection overnight. The reaction solution was filtered through diatomaceous earth, concentrated under reduced pressure, and purified by rapid silica gel column to obtain a yellow solid I-1 (53 mg, 71%). 1 H NMR (400 MHz, DMSO- d 6) δ 10.49 (s, 1H), 9.05 (d, J = 2.2 Hz, 1H),8.69 (s, 1H), 8.57 (brs, 2H), 8.27 (d, J = 2.0 Hz, 1H), 8.20 (dd, J= 9.1, 2.2Hz, 1H), 7.96 (d, J = 8.5 Hz, 1H), 7.93 – 7.87 (m, 2H), 7.12 (s, 2H), 6.64 (s,1H), 3.93 (s, 6H), 3.74 (s, 3H), 2.61 (s, 3H). Example 2: Preparation of N-(1-methyl-1H-pyrazol-4-yl)-7-(3,4,5-trimethoxyphenyl)quinoxaline-2-amine (I-2) Synthesis route:

[0021] Synthesis method: Step 1, 7-bromo-2-chloroquinoxaline (438 mg, 1.8 mmol), 1-methyl-1H-pyrazol-4-amine (146 mg, 1.5 mmol), tris(dibenzylideneacetone)dipalladium (69 mg, 0.075 mmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (93 mg, 0.15 mmol), sodium carbonate (477 mg, 4.5 mmol), and 4 mL of 1,4-dioxane were placed in a 25 mL round-bottom flask, argon was replaced three times, and the reaction was carried out at 100° C. for 22 h under argon protection. The reaction solution was filtered through diatomaceous earth, concentrated under reduced pressure, and purified by a rapid silica gel column to obtain a light yellow solid A4 (51 mg, 11%). 1 H NMR (400 MHz, DMSO- d 6) δ 10.15 (s, 1H), 8.44 (s, 1H), 8.42 (s, 1H), 7.98 (d, J = 2.3 Hz, 1H), 7.74 (d, J = 8.7 Hz, 1H),7.62 (s, 1H), 7.51 (dd, J = 8.6, 2.2 Hz, 1H), 3.87 (s, 3H). Step 2, take A4 (51 mg, 0.17 mmol), 3,4,5-trimethoxyphenylboronic acid (54 mg, 0.26 mmol), bistriphenylphosphine palladium dichloride (5.6 mg, 0.008 mmol), sodium carbonate (51 mg, 0.48 mmol), 3 mL 1,4-dioxane and 0.2 mL water into a 25 mL round-bottom flask, replace argon three times, and react at 100 ° C for 15 hours under argon protection. The reaction solution is filtered through diatomaceous earth, concentrated under reduced pressure, and purified by rapid silica gel column to obtain yellow solid I-2 (50 mg, 76%).1 H NMR (400 MHz, DMSO- d 6) δ 9.98 (s, 1H), 8.42 (s, 1H), 8.38 (s, 1H), 8.02 (s, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.75 (d, J = 8.5 Hz, 1H), 7.69 (s, 1H), 7.08 (s, 2H), 3.92 (s, 6H), 3.89 (s, 3H), 3.73 (s, 3H). Example 3: Preparation of N 1 ,N 1 -Dimethyl-N 4 -(7-(3,4,5-trimethoxyphenyl)quinoxaline-2-yl)benzene-1,4-diamine (I-3) Structural formula:

[0022] Referring to the synthesis method of Example 2, the raw material 1-methyl-1H-pyrazole-4-amine was replaced with 4-amino-N,N-dimethylaniline of equal molar mass, and the yield was 83%. 1 H NMR (400 MHz, DMSO- d 6) δ 9.81 (s, 1H), 8.47 (s, 1H), 7.83 – 7.67 (m, 4H), 7.50 (dd, J = 8.7, 2.2 Hz, 1H), 6.82 – 6.74 (m, 2H), 2.89 (s, 6H). 1 H NMR (400 MHz, DMSO- d 6) δ 9.67 (s, 1H), 8.45 (s, 1H), 7.90 – 7.77(m, 4H), 7.74 (dd, J = 8.5, 2.0 Hz, 1H), 7.06 (s, 2H), 6.82 – 6.77 (m, 2H), 3.91 (s, 6H), 3.72 (s, 3H), 2.88 (s, 6H). Example 4: Preparation of N-(4-amino-2-methylquinolin-6-yl)-2-(4-(5-fluoro-2-(3,4,5-trimethyloxyphenyl)pyrimidin-4-yl)piperazin-1-yl)acetamide (I-4) Synthesis route:

[0023] Synthesis method: Step 1, take the raw materials 2-(piperazine-1-yl)acetic acid (B1, 0.72 g, 5 mmol), 2,4-dichloro-5-fluoropyrimidine (B2, 1.67 g, 10 mmol), DIPEA (1.73 mL, 10 mmol) and 9 mL of ethanol into a 25 mL round-bottom flask, stir at room temperature for 12 hours, filter and dry to obtain a white solid B3 (0.71 g, 52%). 1 H NMR (400 MHz, Chloroform- d ) δ 7.97 (d, J = 6.0 Hz, 1H), 3.91 (t, J = 5.1 Hz, 4H), 3.28 (s, 2H), 2.82 –2.76 (m, 4H). Step 2, take B3 (165 mg, 0.6 mmol) and dissolve it in 4 ml of DMF, add DIPEA (0.31 ml, 1.8 mmol), slowly add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 239 mg, 0.63 mmol), stir at room temperature for half an hour, slowly add A2 (104 mg, 0.6 mmol), stir at room temperature for 6 hours, concentrate the reaction solution under reduced pressure, and purify it by rapid silica gel column to obtain white solid B4 (167 mg, 65%). 1 H NMR (400 MHz, DMSO- d 6) δ 10.07 (s, 1H), 8.41 (d, J = 2.4 Hz, 1H), 8.22 (d, J = 6.5 Hz, 1H),7.91 (dd, J = 9.1, 2.2 Hz, 1H), 7.84 – 7.57 (m, 3H), 3.83 (t, J = 4.9 Hz, 4H), 3.27 (s, 2H), 2.70 (t, J = 5.0 Hz, 4H), 2.50 (s, 3H). Step 3, take B4 (167 mg, 0.39 mmol), 3,4,5-trimethoxyphenylboronic acid (124 mg, 0.58 mmol), bistriphenylphosphine palladium dichloride (14 mg, 0.02 mmol), potassium carbonate (166 mg, 1.2 mmol), 4 mL 1,4-dioxane and 0.2 mL water into a 25 mL round-bottom flask, replace argon three times, and react at 100 ° C for 15 hours under argon protection. The reaction solution is filtered through diatomaceous earth, concentrated under reduced pressure, and purified by rapid silica gel column to obtain yellow solid I-4 (162 mg, 74%). 1 H NMR (400 MHz, Chloroform- d ) δ 9.33 (s, 1H), 8.52 (d, J = 2.3 Hz, 1H), 8.18 (d, J =6.3 Hz, 1H), 7.93 (d, J = 9.0 Hz, 1H), 7.59 (s, 2H), 7.49 (dd, J = 9.0, 2.2 Hz,1H), 6.50 (s, 1H), 5.12 (s, 2H), 3.99 (t, J = 5.0 Hz, 4H), 3.95 (s, 6H), 3.90 (s, 3H), 3.28 (s, 2H), 2.83 (t, J = 4.9 Hz, 4H), 2.58 (s, 3H). Example 5: Preparation of 2-methyl-N 6 -(3-(3,4,5-trimethoxyphenyl)quinoxaline-6-yl)quinoline-4,6-diamine (I-5) Synthesis route:

[0024] Synthesis method: Step 1, take 7-bromo-2-chloroquinoxaline (243 mg, 1 mmol), 3,4,5-trimethoxyphenylboronic acid (212 mg, 1 mmol), bistriphenylphosphine palladium dichloride (35 mg, 0.05 mmol), sodium carbonate (318 mg, 3 mmol), 5 mL of 1,4-dioxane and 0.5 mL of water into a 25 mL round-bottom flask, replace argon three times, and react at 100 ° C for 15 hours under argon protection, and filter to obtain a yellow solid C1 (330 mg, 88%). 1 H NMR (400 MHz, DMSO- d6) δ 9.69 (s, 1H), 8.37 (d, J = 2.2 Hz, 1H), 8.06 (d, J = 8.8 Hz, 1H), 7.96 (dd, J = 8.8, 2.2 Hz, 1H), 7.65(s, 2H), 3.94 (s, 6H), 3.77 (s, 3H). Step 2, C1 (250 mg, 0.67 mmol), A2 (100 mg, 0.58 mmol), Pd2dba3 (23 mg, 0.025 mmol), 2-dicyclohexylphospho-2',4',6'-triisopropylbiphenyl (Xphos, 24 mg, 0.05 mmol), cesium carbonate (490 mg, 1.5 mmol), and 4 mL 1,4-dioxane were placed in a 25 mL round-bottom flask, argon was replaced three times, and the reaction was carried out at 100 ° C for 20 h under argon protection. The reaction solution was filtered through diatomaceous earth, concentrated under reduced pressure, and purified by rapid silica gel column to obtain a yellow solid I-5 (27 mg, 10%). 1 H NMR (400 MHz, DMSO- d 6) δ 9.27 (s, 1H), 8.96 (s, 1H), 7.99 (d, J =2.3 Hz, 1H), 7.94 (d, J = 9.1 Hz, 1H), 7.73 (d, J = 8.9 Hz, 1H), 7.60 – 7.51 (m,4H), 7.41 (d, J = 2.5 Hz, 1H), 6.67 (brs, 2H), 6.44 (s, 1H), 3.90 (s, 6H), 3.74 (s, 3H), 2.42 (s, 3H). Example 6: Preparation of 1-(4-amino-2-methylquinolin-6-yl)-3-(3',4',5'-trimethoxy-[1,1'-diphenyl]-3-yl)urea (I-6) Synthesis route:

[0025] Synthesis method: Step 1, take A2 (86 mg, 0.5 mmol), 3-bromophenyl isocyanate (D1, 0.18 ml, 1.5 mmol), 2.5 ml acetonitrile, and 1.5 ml DMF and place them in a 25 ml round-bottom flask. Stir at room temperature for 12 hours, filter, and wash the filter cake with ethyl acetate and methanol several times to obtain a white solid D2 (176 mg, 95%). 1 H NMR (400 MHz, DMSO- d 6) δ 9.61 (s, 1H), 9.54 (s, 1H), 8.63 (brs, 2H), 8.38 (d, J = 2.1 Hz, 1H), 7.95 (t, J = 2.0 Hz, 1H),7.89 (dd, J = 9.1, 2.2 Hz, 1H), 7.85 – 7.79 (m, 2H), 7.36 – 7.29 (m, 2H), 6.56 (s, 1H), 2.57 (s, 3H). Step 2, take D2 (100 mg, 0.27 mmol), 3,4,5-trimethoxyphenylboronic acid (92 mg, 0.43 mmol), tetrakis(triphenylphosphine)palladium (16 mg, 0.014 mmol), potassium carbonate (112 mg, 0.81 mmol), 4 mL 1,4-dioxane and 0.2 mL water into a 25 mL round-bottom flask, replace argon three times, and react at 100 ° C for 15 hours under argon protection. The reaction solution is filtered through diatomaceous earth, concentrated under reduced pressure, and purified by rapid silica gel column to obtain white solid I-6 (69 mg, 56%). 1 H NMR (400MHz, DMSO- d 6) δ 9.25 (s, 1H), 9.18 (s, 1H), 8.62 (s, 2H), 8.36 (d, J = 2.2 Hz,1H), 7.95 (dd, J = 9.1, 2.2 Hz, 1H), 7.81 (d, J = 9.1 Hz, 1H), 7.71 (t, J = 2.0 Hz,1H), 7.54 (dt, J = 8.3, 1.3 Hz, 1H), 7.39 (t, J = 7.9 Hz, 1H), 7.31 (dt, J= 7.7,1.4 Hz, 1H), 6.87 (s, 2H), 6.56 (s, 1H), 3.87 (s, 6H), 3.70 (s, 3H), 2.57 (s, 3H). Example 7: Preparation of N-(4-amino-2-methylquinolin-6-yl)-2-(3,4,5-trimethoxyphenyl)benzo[d]thiazole-6-carboxamide (I-7) Synthesis route:

[0026] Synthesis method: Step 1, 2-chloro-6-benzothiazolecarboxylic acid (214 mg, 1 mmol) and 3,4,5-trimethoxyphenylboronic acid (318 mg, 1.5 mmol), tetrakis(triphenylphosphine)palladium (35 mg, 0.05 mmol), sodium carbonate (424 mg, 4 mmol), 5 mL of 1,4-dioxane and 0.3 mL of water were placed in a 25 mL round-bottom flask, argon was replaced three times, and the reaction was carried out at 100 ° C for 8 hours under argon protection. The reaction solution was diluted with water (20 mL), 4 N HCl was added to adjust the pH to neutral, and the reaction solution was filtered, and the filter cake was washed with water, and then with ethyl acetate and methanol to obtain a white solid product E1 (62 mg, 18%). 1 H NMR (400 MHz, DMSO- d 6) δ 13.20 (s, 1H), 8.77 (d, J = 1.7 Hz, 1H), 8.13 (d, J = 8.6 Hz, 1H), 8.07 (dd, J = 8.6, 1.7Hz, 1H), 7.38 (s, 2H), 3.93 (s, 6H), 3.77 (s, 3H). Step 2, take E1 (165 mg, 0.6 mmol) and dissolve it in 4 ml DMF, add DIPEA (0.1 ml, 0.54 mmol), slowly add HATU (72 mg, 0.19 mmol), stir at room temperature for half an hour, slowly add A2 (31 mg, 0.18 mmol), stir at room temperature overnight, concentrate the reaction solution under reduced pressure, and purify by rapid silica gel column to obtain yellow solid I-7 (31 mg, 34%). 1 H NMR (400 MHz, DMSO- d 6) δ 10.90 (s, 1H), 8.86 (d, J = 1.7 Hz, 1H), 8.80(d,J = 2.1 Hz, 1H), 8.56 (brs, 2H), 8.23 ​​(d, J = 8.6 Hz, 1H), 8.19 (dd, J = 8.5,1.7 Hz, 1H), 8.07 (dd, J = 9.0, 2.1 Hz, 1H), 7.93 (d, J = 9.0 Hz, 1H), 7.41 (s,2H), 6.60 (s, 1H), 3.94 (s, 6H), 3.78 (s, 3H), 2.59 (s, 3H). Example 8: Preparation of N-(4-amino-2-methylquinolin-6-yl)-2-(3',4',5'-trimethoxy-[1,1'-diphenyl]-3-yl)acetamide (I-8) Synthesis route:

[0027] Synthesis method: Step 1, take m-bromophenylacetic acid (F1, 215 mg, 1 mmol) and 3,4,5-trimethoxyphenylboronic acid (339 mg, 1.6 mmol), tetrakis(triphenylphosphine)palladium (35 mg, 0.05 mmol), cesium carbonate (1300 mg, 4 mmol), 5 mL of 1,4-dioxane and 0.5 mL of water in a 25 mL round-bottom flask, replace argon three times, and react at 100°C for 17 hours under argon protection. The reaction solution is diluted with water, 4 N HCl is added to adjust the pH to neutral, and extracted with dichloromethane. The organic phase is collected and then concentrated under reduced pressure. Purification by rapid silica gel column obtains white solid F2 (120 mg, 40%). 1 H NMR (400 MHz, DMSO- d 6) δ 12.37(s, 1H), 7.59 – 7.52 (m, 1H), 7.42 – 7.34 (m, 1H), 7.24 (dt, J = 7.5, 1.4 Hz,1H), 6.89 (s, 2H), 3.86 (s, 6H), 3.69 (s, 3H), 3.65 (s, 2H). Step 2, take F2 (120 mg, 0.4 mmol) and dissolve it in 4 ml DMF, add DIPEA (0.26 ml, 1.5 mmol), slowly add HATU (167 mg, 0.44 mmol), stir at room temperature for half an hour, slowly add A2 (69 mg, 0.4 mmol), stir at room temperature for 4 hours, concentrate the reaction solution under reduced pressure, and purify it by rapid silica gel column to obtain yellow solid I-8 (71 mg, 39%). 1 H NMR (400 MHz, DMSO- d 6) δ 10.33 (s, 1H), 8.24 (s, 1H), 7.66 (t, J =1.8 Hz, 1H), 7.60 (s, 2H), 7.59 – 7.55 (m, 1H), 7.42 (t, J = 7.6 Hz, 1H), 7.36(d, J = 7.6 Hz, 1H), 6.90 (s, 2H), 6.40 (s, 1H), 6.37 (s, 2H), 3.86 (s, 6H), 3.77 (s, 2H), 3.69 (s, 3H), 2.37 (s, 3H). Example 9: Preparation of N-(4-amino-2-methylquinolin-6-yl)-2-((3',4',5'-trimethoxy-[1,1'-diphenyl]-3-yl)oxy)acetamide (I-9) Structural formula:

[0028] Referring to the synthesis method of Example 8, the raw material m-bromophenylacetic acid was replaced with 2-(3-bromophenoxy)acetic acid of equal molar mass, and the yield was 43%. 1 H NMR (400 MHz, DMSO- d 6) δ 10.67 (s, 1H), 8.64 (d, J = 2.1 Hz,1H), 8.55 (brs, 2H), 7.96 (dd, J = 9.1, 2.1 Hz, 1H), 7.88 (d, J = 9.1 Hz, 1H),7.40 (t, J= 7.9 Hz, 1H), 7.34 – 7.27 (m, 2H), 7.06 – 7.00 (m, 1H), 6.89 (s,2H), 6.57 (s, 1H), 4.90 (s, 2H), 3.83 (s, 6H), 3.69 (s, 3H), 2.57 (s, 3H). Example 10: Preparation of N-(4-amino-2-methylquinolin-6-yl)-2-(2-(4-(5-fluoro-2-(3,4,5-trimethoxyphenyl)pyrimidin-4-yl)piperazin-1-yl)acetamido)acetamide (I-10) Synthesis route:

[0029] Synthesis method: Step 1, take B1 (275 mg, 1 mmol) and dissolve it in 5 ml of dichloromethane, add DIPEA (0.42 ml, 3 mmol), slowly add HATU (399 mg, 1.05 mmol), stir at room temperature for half an hour, slowly add glycine tert-butyl ester (0.15 ml, 1.1 mmol), stir at room temperature overnight, concentrate the reaction solution under reduced pressure, and purify it by rapid silica gel column to obtain white oily solid G1 (320 mg, 82%). 1 H NMR (400 MHz, Chloroform- d ) δ 7.94 (d, J = 6.1 Hz,1H), 7.51 (s, 1H), 3.99 (d, J = 5.5 Hz, 2H), 3.93 – 3.84 (m, 4H), 3.11 (s, 2H), 2.73 – 2.64 (m, 4H), 1.48 (s, 9H). Step 2, G1 (252 mg, 0.65 mmol), 3,4,5-trimethoxyphenylboronic acid (220 mg, 1.04 mmol), bistriphenylphosphine palladium dichloride (23 mg, 0.03 mmol), potassium carbonate (269 mg, 1.95 mmol), 4 mL 1,4-dioxane and 0.2 mL water were placed in a 25 mL round-bottom flask, argon was replaced three times, and the reaction was carried out at 100 ° C for 12 hours under argon protection. The reaction solution was filtered through diatomaceous earth, concentrated under reduced pressure, and purified by rapid silica gel column to obtain oily solid G2 (300 mg, 89%). 1 H NMR (400 MHz, DMSO- d 6) δ 8.35 (d, J= 6.6 Hz, 1H), 8.16 (t, J = 6.1 Hz, 1H), 7.58 (s,2H), 3.89 – 3.82 (m, 10H), 3.77 (d, J = 6.1 Hz, 2H), 3.72 (s, 3H), 3.04 (s,2H), 2.64 (t, J = 4.9 Hz, 4H), 1.41 (s, 9H). Step 3, G2 (300 mg, 0.58 mmol) was dissolved in 1 ml of dichloromethane, 1 ml of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 2 hours, and then concentrated under reduced pressure to obtain a yellow oily solid G3 (232 mg, 95%). 1 H NMR (400 MHz, DMSO- d 6) δ 8.97 (t, J = 5.8 Hz, 1H), 8.47 (d, J = 6.3 Hz, 1H), 7.59 (s, 2H), 4.89– 4.35 (m, 4H), 3.90 (d, J = 5.8 Hz, 2H), 3.86 (s, 6H), 3.73 (s, 3H), 3.67 –3.22 (m, 6H). Step 4, G3 (275 mg, 1 mmol) was dissolved in 2 ml DMF, DIPEA (0.35 ml, 2 mmol) was added, HATU (200 mg, 0.53 mmol) was slowly added, and after stirring at room temperature for half an hour, A2 (86 mg, 0.5 mmol) was slowly added. The reaction was stirred at room temperature overnight, and the reaction solution was concentrated under reduced pressure. The white solid I-10 (110 mg, 36%) was obtained by rapid silica gel column purification. 1 HNMR (400 MHz, DMSO- d 6) δ 10.15 (s, 1H), 8.35 (d, J = 6.7 Hz, 1H), 8.25 (s, 1H), 8.16 (t, J = 6.0 Hz, 1H), 7.68 – 7.60 (m, 2H), 7.58 (s, 2H), 6.73 (brs, 2H), 6.45 (s, 1H), 4.01 (d, J= 5.9 Hz, 2H), 3.92 – 3.79 (m, 10H), 3.72 (s, 3H), 3.09 (s, 2H), 2.68 (t, J = 4.9 Hz, 4H), 2.41 (s, 3H). Example 11: Preparation of N-(4-amino-2-methylquinolin-6-yl)-7-(3,4,5-trimethoxyphenyl)quinoline-2-carboxamide (I-11) Structural formula:

[0030] Referring to the synthesis method of Example 7, the raw material 2-chloro-6-benzothiazolecarboxylic acid was replaced with 7-bromoquinoline-2-carboxylic acid of equal molar mass, and the yield was 61%. 1 H NMR (400 MHz, DMSO- d 6) δ 11.13 (s, 1H), 8.92 – 8.61 (m,4H), 8.55 – 8.51 (m, 1H), 8.38 (dd, J = 9.1, 2.2 Hz, 1H), 8.28 (d, J = 8.4 Hz,1H), 8.24 (d, J = 8.6 Hz, 1H), 8.20 (dd, J = 8.6, 1.8 Hz, 1H), 7.97 (d, J = 9.1 Hz,1H), 7.18 (s, 2H), 6.63 (s, 1H), 3.94 (s, 6H), 3.75 (s, 3H), 2.62 (s, 3H). Example 12: Preparation of N-(4-amino-2-methylquinolin-6-yl)-3',4',5'-trimethoxy-[1,1'-diphenyl]-3-carboxamide (I-12) Structural formula:

[0031] Referring to the synthesis method of Example 8, the raw material m-bromophenylacetic acid was replaced with m-bromobenzoic acid of equal molar mass, and the yield was 47%. 1 H NMR (400 MHz, DMSO- d 6) δ 10.92 (s, 1H), 8.83 (d, J = 2.1 Hz, 1H), 8.70(brs, 2H), 8.33 (t, J = 1.8 Hz, 1H), 8.12 (dd,J = 9.1, 2.1 Hz, 1H), 8.03 – 7.98(m, 1H), 7.98 – 7.91 (m, 2H), 7.65 (t, J = 7.7 Hz, 1H), 7.06 (s, 2H), 6.61 (s,1H), 3.90 (s, 6H), 3.72 (s, 3H), 2.60 (s, 3H). Example 13: 2-Methyl-N 6 -(2-(3,4,5-trimethoxyphenyl)thieno[3,2-d]pyrimidin-4-yl)quinoline-4,6-diamine (I-13) Synthesis route:

[0032] Synthesis method: Step 1, take raw material A2 (86 mg, 0.5 mmol), 2,4-dichlorothieno[3,2-d]pyrimidine (H1, 205 mg, 1 mmol), DIPEA (0.17 mL, 1 mmol) and 3 mL of ethanol and put them in a 25 mL round-bottom flask, stir and react at 85°C for 6 hours, filter, wash the filter cake with ethyl acetate and methanol, and dry to obtain white solid H2 (150 mg, 88%). 1 H NMR (400 MHz, DMSO- d 6) δ 10.59 (s, 1H), 8.40 (d, J = 2.2 Hz, 1H), 8.29 (d, J = 5.4 Hz, 1H), 8.23 ​​(brs, 2H), 7.98 (dd, J = 9.0, 2.1 Hz, 1H), 7.90 (d, J = 9.0 Hz, 1H), 7.44(d, J = 5.4 Hz, 1H), 6.59 (s, 1H), 2.58 (s, 3H). Step 2, take H2 (147 mg, 0.43 mmol), 3,4,5-trimethoxyphenylboronic acid (137 mg, 0.65 mmol), bistriphenylphosphine palladium dichloride (18 mg, 0.03 mmol), potassium carbonate (207 mg, 1.5 mmol), 4 mL 1,4-dioxane and 0.3 mL water into a 25 mL round-bottom flask, replace argon three times, and react at 100 ° C for 14 hours under argon protection. The reaction solution is filtered through diatomaceous earth and concentrated under reduced pressure. Purification by rapid silica gel column gives white solid I-13 (124 mg, 61%). 1 H NMR (400 MHz, DMSO- d 6) δ 9.99 (s, 1H), 8.35 (d, J = 2.2 Hz, 1H), 8.22 (d, J = 5.4 Hz,1H), 7.94 (dd, J = 8.9, 2.2 Hz, 1H), 7.78 – 7.70 (m, 3H), 7.55 (d, J = 5.4 Hz,1H), 6.77 (brs, 2H), 6.47 (s, 1H), 3.81 (s, 6H), 3.71 (s, 3H), 2.44 (s, 3H). Example 14: Preparation of N 6 -(2-(Benzo[d][1,3]dioxol-5-yl)thieno[3,2-d]pyrimidin-4-yl)-2-methylquinoline-4,6-diamine (I-14) Structural formula:

[0033] Referring to the synthesis method of Example 13, the raw material 3,4,5-trimethoxyphenylboronic acid was replaced with 3,4-methyleneoxyphenylboronic acid of equal molar mass, and the yield was 70%. 1 H NMR (400 MHz, DMSO- d 6) δ 10.28 (s, 1H), 8.88 –8.47 (m, 3H), 8.28 (d, J = 5.4 Hz, 1H), 8.16 (dd, J = 9.0, 2.1 Hz, 1H), 8.06 –7.89 (m, 2H), 7.81 (d, J = 1.7 Hz, 1H), 7.54 (d, J = 5.4 Hz, 1H), 6.98 (d, J= 8.2Hz, 1H), 6.64 (s, 1H), 6.09 (s, 2H), 2.63 (s, 3H). Example 15: Preparation of N 6 -(2-(1H-indol-5-yl)thieno[3,2-d]pyrimidin-4-yl)-2-methylquinoline-4,6-diamine (I-15) Structural formula:

[0034] Referring to the synthesis method of Example 13, the raw material 3,4,5-trimethoxyphenylboronic acid was replaced with 5-indoleboric acid of equal molar mass, and the yield was 23%. 1 H NMR (400 MHz, DMSO- d 6) δ 11.51 (s, 1H), 9.01 – 8.61 (m,4H), 8.43 (s, 1H), 8.21 (dd, J = 9.0, 2.1 Hz, 1H), 8.14 (d, J = 8.6 Hz, 1H), 8.04(d, J = 9.0 Hz, 1H), 7.70 (d, J = 5.3 Hz, 1H), 7.56 – 7.43 (m, 2H), 6.69 (s, 1H), 6.56 (s, 1H), 2.65 (s, 3H). Example 16: Preparation of N 6 -(2-(3,4-dihydroisoquinolin-2(1H)-yl)thieno[3,2-d]pyrimidin-4-yl)-2-methylquinoline-4,6-diamine (I-16) Structural formula:

[0035] Referring to the synthesis method of Example 13, the raw material 3,4,5-trimethoxyphenylboronic acid was replaced with 1,2,3,4-tetrahydroisoquinoline of equal molar mass, and the yield was 90%. 1 H NMR (400 MHz, DMSO- d 6) δ 9.94 (s, 1H), 8.90 – 8.70 (m, 2H), 8.52 (s, 1H), 8.13 – 8.02 (m, 2H), 7.94 (d, J = 9.1 Hz, 1H), 7.23 (d, J= 5.3 Hz, 1H), 7.21 – 7.11 (m, 4H), 6.64 (s, 1H), 4.85 (s, 2H), 3.96 (t, J =5.8 Hz, 2H), 2.86 (t, J = 5.9 Hz, 2H), 2.63 (s, 3H). Example 17: Preparation of N 6 -(2-(4-methoxypiperidin-1-yl)thieno[3,2-d]pyrimidin-4-yl)-2-methylquinoline-4,6-diamine (I-17) Structural formula:

[0036] Referring to the synthesis method of Example 13, the raw material 3,4,5-trimethoxyphenylboronic acid was replaced with 4-methoxypiperidine of equal molar mass, and the yield was 54%. 1 H NMR (400 MHz, DMSO- d 6) δ 9.51 (s, 1H), 8.28 (d, J = 2.2Hz, 1H), 7.96 (d, J = 5.4 Hz, 1H), 7.70 (dd, J = 9.0, 2.2 Hz, 1H), 7.63 (d, J = 8.9Hz, 1H), 7.12 (d, J = 5.4 Hz, 1H), 6.47 – 6.36 (m, 3H), 4.22 (dt, J = 13.1, 4.7Hz, 2H), 3.43 – 3.37 (m, 1H), 3.29 – 3.19 (m, 5H), 2.40 (s, 3H), 1.90 – 1.80(m, 2H), 1.44 – 1.32 (m, 2H). Example 18: Preparation of 2-methyl-N 6 -(2-(4-morpholinylphenyl)thieno[3,2-d]pyrimidin-4-yl)quinoline-4,6-diamine (I-18) Structural formula:

[0037] Referring to the synthesis method of Example 13, the raw material 3,4,5-trimethoxyphenylboronic acid was replaced with 4-(4-morpholinyl)phenylboronic acid of equal molar mass, and the yield was 57%. 1 H NMR (400 MHz, DMSO- d6) δ 9.89 (brs, 1H), 8.35 (d, J = 2.3 Hz, 1H), 8.27 (d, J = 8.9 Hz, 2H), 8.13 (d, J = 5.4 Hz, 1H), 7.85 (dd, J =9.0, 2.3 Hz, 1H), 7.70 (d, J = 9.0 Hz, 1H), 7.45 (d, J = 5.4 Hz, 1H), 6.99 (d, J =9.1 Hz, 2H), 6.50 (s, 2H), 6.46 (s, 1H), 3.79 – 3.69 (m, 4H), 3.23 – 3.17 (m,4H), 2.43 (s, 3H). Example 19: Preparation of N 6 -(2-(6-methoxypyridin-3-yl)thieno[3,2-d]pyrimidin-4-yl)-2-methylquinoline-4,6-diamine (I-19) Structural formula:

[0038] Referring to the synthesis method of Example 13, the raw material 3,4,5-trimethoxyphenylboronic acid was replaced with 6-methoxypyridine-3-boric acid of equal molar mass, and the yield was 68%. 1 H NMR (400 MHz, DMSO- d 6) δ 10.35 (s, 1H), 9.07 (d, J = 2.3 Hz, 1H), 8.74 – 8.61 (m, 3H), 8.55 (dd, J = 8.8, 2.4 Hz, 1H), 8.31 (d, J =5.5 Hz, 1H), 8.18 (dd, J = 9.1, 2.1 Hz, 1H), 8.01 (d, J = 8.8 Hz, 1H), 7.57 (d, J =5.4 Hz, 1H), 6.90 (d, J = 8.7 Hz, 1H), 6.65 (s, 1H), 3.91 (s, 3H), 2.63 (s, 3H). Example 20: Preparation of 2-methyl-N 6-(2-(3,4,5-trimethoxyphenyl)benzo[d]thiazol-6-yl)quinoline-4,6-diamine (I-20) Structural formula:

[0039] Referring to the synthesis method of Example 5, the raw material 7-bromo-2-chloroquinoxaline was replaced with 6-bromo-2-chlorobenzothiazole of equal molar mass, and the yield was 28%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.81 (s, 1H), 7.94 (d, J = 8.8Hz, 1H), 7.90 – 7.83 (m, 2H), 7.71 (d, J = 9.0 Hz, 1H), 7.59 – 7.38 (m, 3H), 7.34 – 7.25 (m, 3H), 6.47 (s, 1H), 3.91 (s, 6H), 3.75 (s, 3H), 2.47 (s, 3H). Example 21: Preparation of N 6 -(2-(Benzo[d][1,3]dioxol-5-yl)benzo[d]thiazol-6-yl)-2-methylquinoline-4,6-diamine (I-21) Structural formula:

[0040] Referring to the synthesis method of Example 5, the raw material 3,4,5-trimethoxyphenylboronic acid was replaced with 3,4-methyleneoxyphenylboronic acid of equal molar mass, and the yield was 32%. 1 H NMR (400 MHz, DMSO- d 6) δ 9.08 (s, 1H), 8.57 (brs,2H), 7.98 – 7.89 (m, 3H), 7.87 (d, J = 8.9 Hz, 1H), 7.68 (dd, J = 9.1, 2.3 Hz,1H), 7.61 – 7.51 (m, 2H), 7.33 (dd, J = 8.8, 2.3 Hz, 1H), 7.09 (d, J = 8.0 Hz,1H), 6.55 (s, 1H), 6.16 (s, 2H), 2.57 (s, 3H). Example 22: Preparation of N 6-(2-(3,4-dihydroisoquinolin-2(1H)-yl)benzo[d]thiazol-6-yl)-2-methylquinoline-4,6-diamine (I-22) Synthesis route:

[0041] Synthesis method: Step 1, 6-bromo-2-chlorobenzothiazole (J1, 497 mg, 2 mmol) and 1,2,3,4-tetrahydroisoquinoline (J2, 0.5 ml, 4 mmol) were dissolved in 6 ml of ethanol, DIPEA (0.69 ml, 4 mmol) was added, and the mixture was stirred at 85°C for overnight. The mixture was cooled to room temperature, and a large amount of solid precipitated. The mixture was filtered and washed with a small amount of ethyl acetate to obtain a white solid J3 (560 mg, 81%). 1 H NMR (400MHz, Chloroform- d ) δ 7.73 (d, J = 1.9 Hz, 1H), 7.44 (d, J = 8.5 Hz, 1H), 7.39(dd, J = 8.6, 2.0 Hz, 1H), 7.25 – 7.18 (m, 4H), 4.81 (s, 2H), 3.86 (t, J = 5.9Hz, 2H), 3.03 (t, J = 5.9 Hz, 2H). Step 2, J3 (224 mg, 0.65 mmol), A2 (86 mg, 0.5 mmol), Pd2dba3 (23 mg, 0.025 mmol), Xphos (24 mg, 0.05 mmol), cesium carbonate (490 mg, 1.5 mmol), and 4 mL of 1,4-dioxane were placed in a 25 mL round-bottom flask, argon was replaced three times, and the reaction was carried out at 100 ° C for 15 h under argon protection. The reaction solution was filtered through diatomaceous earth, concentrated under reduced pressure, and purified by flash silica gel column to obtain yellow solid I-5 (92 mg, 42%). 1 H NMR (400 MHz, DMSO- d 6) δ8.62 (s, 1H), 8.40 (brs, 2H), 7.74 – 7.68 (m, 2H), 7.66 (d, J = 2.4 Hz, 1H),7.56 (dd, J = 9.1, 2.3 Hz, 1H), 7.47 (d, J= 8.5 Hz, 1H), 7.33 – 7.29 (m, 1H), 7.27 – 7.20 (m, 3H), 7.15 (dd, J = 8.6, 2.3 Hz, 1H), 6.49 (s, 1H), 4.77 (s,2H), 3.82 (t, J = 6.0 Hz, 2H), 2.99 (t, J = 5.9 Hz, 2H), 2.54 (s, 3H). Example 23: Preparation of N 6 -(2-(4-methoxypiperidin-1-yl)benzo[d]thiazol-6-yl)-2-methylquinoline-4,6-diamine (I-23) Structural formula:

[0042] Referring to the synthesis method of Example 22, the raw material 1,2,3,4-tetrahydroisoquinoline was replaced with 4-methoxypiperidine of equal molar mass, and the yield was 48%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.62 (s, 1H), 8.41 (brs, 2H), 7.71 (d, J = 9.1 Hz, 1H), 7.66 (t, J = 2.8 Hz, 2H), 7.56 (dd, J = 9.2, 2.3 Hz, 1H),7.42 (d, J = 8.5 Hz, 1H), 7.13 (dd, J = 8.6, 2.3 Hz, 1H), 6.50 (s, 1H), 3.77 (dt, J = 13.2, 5.1 Hz, 2H), 3.48 (tt, J = 7.9, 3.4 Hz, 1H), 3.41 – 3.36 (m, 1H), 3.30 (s, 3H), 2.54 (s, 3H), 1.99 – 1.90 (m, 2H), 1.61 – 1.48 (m, 2H). Example 24: Preparation of (6-((4-amino-2-methylquinolin-6-yl)amino)benzo[d]thiazol-2-yl)-L-phenylalanine tert-butyl ester (I-24) Structural formula:

[0043] Referring to the synthesis method of Example 22, the raw material 1,2,3,4-tetrahydroisoquinoline was replaced with L-phenylalanine tert-butyl ester of equal molar mass, and the yield was 23%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.58 (s, 1H), 8.39 (brs,2H), 8.32 (d, J = 7.7 Hz, 1H), 7.71 (d, J = 9.1 Hz, 1H), 7.62 (d, J = 2.3 Hz, 1H),7.59 (d, J = 2.3 Hz, 1H), 7.55 (dd, J = 9.1, 2.3 Hz, 1H), 7.34 (d, J = 8.5 Hz, 1H),7.33 – 7.28 (m, 4H), 7.27 – 7.21 (m, 1H), 7.08 (dd, J = 8.6, 2.3 Hz, 1H), 6.49(s, 1H), 4.56 (td, J = 8.1, 6.5 Hz, 1H), 3.13 – 3.00 (m, 2H), 2.54 (s, 3H), 1.33 (s, 9H). Example 25: Preparation of 2-methyl-N 6 -(2-(3,4,5-trimethoxyphenyl)thieno[2,3-d]pyrimidin-4-yl)quinoline-4,6-diamine (I-25) Structural formula:

[0044] Referring to the synthesis method of Example 13, the raw material 2,4-dichlorothieno[3,2-d]pyrimidine was replaced with 2,4-dichlorothieno[2,3-d]pyrimidine of equal molar mass, and the yield was 73%. 1 H NMR (400 MHz, DMSO- d 6) δ 10.20 (s,1H), 8.54 (s, 1H), 8.15 (d, J = 9.0 Hz, 1H), 7.93 (d, J = 5.9 Hz, 1H), 7.86 (d, J=9.0 Hz, 1H), 7.82 – 7.53 (m, 5H), 6.54 (s, 1H), 3.81 (s, 6H), 3.71 (s, 3H), 2.53 (s, 3H). Example 26: Preparation of 2-methyl-N 6 -(2-(3,4,5-trimethoxyphenyl)-5,7-dihydrofurano[3,4-d]pyrimidin-4-yl)quinoline-4,6-diamine (I-26) Structural formula:

[0045] Referring to the synthesis method of Example 13, the raw material 2,4-dichlorothieno[3,2-d]pyrimidine was replaced with 2,4-dichloro-5,7-dihydrofurano[3,4-d]pyrimidine of equal molar mass, with a yield of 75%. 1 H NMR (400 MHz, DMSO- d 6) δ9.52 (s, 1H), 8.21 (d, J = 2.3 Hz, 1H), 7.86 (dd, J = 9.0, 2.3 Hz, 1H), 7.69 (d, J = 9.0 Hz, 1H), 7.66 (s, 2H), 6.50 (s, 2H), 6.43 (s, 1H), 5.01 (s, 2H), 4.98 –4.92 (m, 2H), 3.80 (s, 6H), 3.71 (s, 3H), 2.41 (s, 3H). Example 27: Preparation of tert-butyl 4-((4-amino-2-methylquinolin-6-yl)amino)-2-(3,4,5-trimethoxyphenyl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (I-27) Structural formula:

[0046] Referring to the synthesis method of Example 13, the raw material 2,4-dichlorothieno[3,2-d]pyrimidine was replaced with 6-Boc-2,4-dichloro-5,7-dihydropyrrolo[3,4-d]pyrimidine of equal molar mass, with a yield of 60%. 1 H NMR (400 MHz, DMSO- d 6)δ 9.49 (s, 1H), 8.24 (t, J = 2.2 Hz, 1H), 7.88 (dd, J = 9.0, 2.3 Hz, 1H), 7.68(d,J = 9.1 Hz, 1H), 7.65 (s, 2H), 6.48 (s, 2H), 6.44 (s, 1H), 4.64 – 4.49 (m,4H), 3.80 (s, 6H), 3.70 (s, 3H), 2.40 (s, 3H), 1.49 (d, J = 5.4 Hz, 9H). Example 28: Preparation of 2-methyl-N 6 -(2-(3,4,5-trimethoxyphenyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)quinoline-4,6-diamine hydrochloride (I-28) Structural formula:

[0047] Synthesis method: Take 84 mg of I-27 and place it in a 25 ml round-bottom flask, add 2 ml of 2 mol / L hydrochloric acid ethyl acetate solution, stir at room temperature for 4 hours, filter, wash with ethyl acetate several times, and dry to obtain a light yellow solid I-28 (71 mg, 96%). 1 HNMR (400 MHz, DMSO- d 6) δ 10.40 – 10.28 (m, 2H), 10.24 (s, 1H), 8.82 (s, 1H), 8.69 – 8.54 (m, 2H), 8.16 (dd, J = 9.1, 2.1 Hz, 1H), 8.00 (d, J = 9.1 Hz, 1H),7.61 (s, 2H), 6.63 (d, J = 1.4 Hz, 1H), 4.65 – 4.58 (m, 2H), 4.51 (t, J = 5.8 Hz, 2H), 3.78 (s, 6H), 3.71 (s, 3H), 2.61 (s, 3H). Example 29: Preparation of 2-methyl-N 6 -(2-(3,4,5-trimethoxyphenyl)quinazolin-4-yl)quinoline-4,6-diamine (I-29) Structural formula:

[0048] Referring to the synthesis method of Example 13, the raw material 2,4-dichlorothieno[3,2-d]pyrimidine was replaced with 2,4-dichloroquinazoline of equal molar mass, and the yield was 82%. 1 H NMR (400 MHz, DMSO-d 6) δ 10.15 (s, 1H), 8.58 (d, J = 8.3 Hz, 1H), 8.39 (d, J = 2.3 Hz, 1H), 8.00 (dd, J = 9.0, 2.2 Hz, 1H), 7.90 –7.83 (m, 2H), 7.80 – 7.71 (m, 3H), 7.65 – 7.58 (m, 1H), 6.59 (s, 2H), 6.46(s, 1H), 3.80 (s, 6H), 3.71 (s, 3H), 2.43 (s, 3H). Example 30: Preparation of 2-methyl-N 6 -(2-(3,4,5-trimethoxyphenyl)pyrrolo[2,1-f][1,2,4]triazine-4-yl)quinoline-4,6-diamine (I-30) Structural formula:

[0049] Referring to the synthesis method of Example 13, the raw material 2,4-dichlorothieno[3,2-d]pyrimidine was replaced with 2,4-dichloropyrrolo[2,1-f][1,2,4]triazine of equal molar mass, and the yield was 71%. 1 H NMR (400 MHz, DMSO- d 6) δ 10.62(s, 1H), 8.90 – 8.60 (m, 3H), 8.31 (dd, J = 9.1, 2.1 Hz, 1H), 8.02 (d, J = 9.0Hz, 1H), 7.91 (dd, J = 2.6, 1.5 Hz, 1H), 7.55 (s, 2H), 7.31 (dd, J = 4.4, 1.6 Hz,1H), 6.82 (dd, J = 4.4, 2.6 Hz, 1H), 6.65 (s, 1H), 3.81 (s, 6H), 3.71 (s, 3H), 2.63 (s, 3H). Example 31: Preparation of tert-butyl 4-((4-amino-2-methylquinolin-6-yl)amino)-2-(3,4,5-trimethoxyphenyl)-7,8-dihydropyrido[4,3-d]pyrimidine-6(5H)-carboxylate (I-31) Structural formula:

[0050] Referring to the synthesis method of Example 13, the raw material 2,4-dichlorothieno[3,2-d]pyrimidine was replaced with N-Boc-2,4-dichloro-5,7,8-trihydropyrido[4,3-D]pyrimidine (Leyan, product number 1061570) of equal molar mass, with a yield of 34%. 1 HNMR (400 MHz, DMSO- d 6) δ 8.99 (s, 1H), 8.31 (s, 1H), 7.93 (d, J = 9.0 Hz, 1H),7.75 (d, J = 9.0 Hz, 1H), 7.57 (s, 2H), 7.15 (brs, 2H), 6.48 (s, 1H), 4.53 (s,2H), 3.75 (s, 6H), 3.73 – 3.65 (m, 5H), 2.84 (t, J = 5.8 Hz, 2H), 2.47 (s, 3H), 1.48 (s, 9H). Example 32: Preparation of 2-methyl-N 6 -(2-(3,4,5-trimethoxyphenyl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yl)quinoline-4,6-diamine hydrochloride (I-32) Structural formula:

[0051] Referring to the synthesis method of Example 28, the raw material I-28 was replaced with I-31 of equal molar mass, and the yield was 97%. 1 HNMR (400 MHz, DMSO- d 6) δ 10.17 (s, 2H), 9.78 (s, 1H), 8.83 (s, 1H), 8.74 –8.58 (m, 2H), 8.12 (dd, J = 9.0, 2.1 Hz, 1H), 8.00 (d, J = 9.1 Hz, 1H), 7.59 (s,2H), 6.62 (d, J = 1.4 Hz, 1H), 4.29 (t, J = 4.8 Hz, 2H), 3.77 (s, 6H), 3.70 (s, 3H), 3.55 – 3.47 (m, 2H), 3.23 – 3.12 (m, 2H), 2.62 (s, 3H). Example 33 Biological Evaluation Experiment (1) Small molecule assay for PRMT7 enzyme activity 1. Prepare 1× assay buffer (10 mM Tirs-HCl (pH=8.5), 0.01% Tween-20, 0.01% BSA and 1 mM dithiothreitol).

[0052] 2. First, dissolve the compound in DMSO to prepare a stock solution of the compound (20 mM). Use 1× assay buffer to prepare a gradient dilution solution of the compound (compound concentrations: 10 μM, 5 μM, 2.5 μM, 1.25 μM, 0.63 μM, 0.31 μM, 0.16 μM, 0.08 μM, and 0.04 μM), keeping the final concentration of DMSO no more than 1%.

[0053] 3. Prepare a 1× assay buffer containing PRMT7 protein (15 nM, Active Motif, Cat. No.31795) and 3 Solution of H-SAM (0.5 μM, PerkinElmer, Cat. No. NET155V001MC).

[0054] 4. Prepare a biotin-labeled substrate peptide H4 (1-21) (sequence: Ac-SGRGKGGKGLGKGGAKRHRKVGG-K(Biotin), GL Biochem (Shanghai) Ltd., 415263) solution (0.5 μM) in 1× assay buffer.

[0055] 5. First, add 200 nL of the compound gradient dilution solution to each well of the 384-well plate. Then, add 10 μL of the PRMT7 protein and 3 Add 10 μl of 1× assay buffer to the H-SAM solution and incubate at room temperature for 15 min.

[0056] 6. Add 10 μl of substrate peptide H4 solution to each well to initiate the enzyme catalytic reaction and incubate at room temperature for 240 minutes.

[0057] 7. Prepare stop solution: 1 × assay buffer containing SAM (10 mM, Sigma, Cat. No. A7007-100MG).

[0058] 8. Add 10 μl of stop solution to each well to stop the enzyme catalyzed reaction.

[0059] 9. Transfer 25 μl of the above reaction solution to a fast plate and incubate at room temperature for 1 hour.

[0060] 10. Read the signal value of each well using the Microbeta instrument.

[0061] 11. Use GraphPad 8.0 software to fit the inhibition rate-concentration curve according to the signal value and calculate the IC 50 value.

[0062] The results are shown in Table 1 below. It can be seen from the experimental results that the compounds of the present invention have significant inhibitory activity on PRMT7 protein.

[0063] Table 1 Inhibitory activity of the compounds of the present invention on PRMT7 protein

[0064] (2) Experiment on small molecule inhibition of tumor cell proliferation Pancreatic cancer cells PATU-8988 (FH0544), breast cancer cells MDA-MB-231 (FH0213), colorectal cancer cells RKO (FH0030), lung cancer cells A549 (FH0045), ovarian cancer cells SKOV-3 (FH0135), prostate cancer cells DU-145 (FH0197), 22Rv1 (FH0198), VCaP (FH0194) and PC-3 (FH0195), esophageal cancer cells ECA109 (FH0225), liver cancer cells HEPG2 (FH0076), leukemia cells MOLM13 (FH1018), and glioma cells U251 (FH0159) were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum at 37 °C and 5% CO2. All cells were from Shanghai Fuheng Biotechnology Co., Ltd. The cell suspension was diluted to 50,000 cells / mL with complete cell culture medium, and 100 μL of culture medium containing 5,000 cells was inoculated into a 96-well plate. The plate was cultured at 37 °C and 5% CO2 for one day. Then, 100 μL of culture medium containing the test compound with gradient concentrations (10 μM, 5 μM, 2.5 μM, 1.25 μM, 0.63 μM, 0.31 μM, 0.16 μM, 0.08 μM and 0.04 μM) was added to each well. Three replicate wells were set for each dose group, and DMSO was set as a control group. After 48 hours of treatment, 10 μL of CCK-8 solution (10 mg / mL, Share-bio, SB-CCK8) was added to each well, incubated at 37 °C for 4 hours, and the OD value of each well at 450 nm was read using a microplate reader (Bio-Tek SynergyH1). The data were processed using EXCEL software and nonlinear fitting and IC calculation were performed using GraphPad8.0 software. 50 Value. Inhibition rate of cell proliferation = (control group OD 450 - Experimental group OD 450 ) / control group OD 450 *100%. The results are shown in Tables 2 and 3 below: Table 2 Inhibitory activity of the compounds of the present invention on PATU-8988 cell line

[0065] It can be seen from Table 2 that most of the compounds of the present invention have strong proliferation inhibition activity on tumor cells.

[0066] Table 3 Inhibitory activity of compound I-13 of the present invention on various cancer cell lines

[0067] As can be seen from Table 3, Example 13 (I-13) of the present invention has significant killing effect on various tumor cell lines, a wide therapeutic range, and good anti-tumor therapeutic potential.

Claims

1. A compound for inhibiting PRMT7 protein activity or a pharmaceutically acceptable salt thereof, characterized in that: The general formula of the compound is shown in any one of formulas (II) to (V): ； Wherein, when X1 and X2 are independently selected from sulfur atoms, CH; R1 is selected from the following groups: ； Wherein, R2, R3, R4, R5, and R6 are independently selected from methoxy, ethyl, and hydrogen; A and B are independently selected from CH or N.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: When X1 is a sulfur atom, X2 is CH; when X1 is CH, X2 is a sulfur atom; R1 is selected from the following groups: 。 3. A compound for inhibiting PRMT7 protein activity or a pharmaceutically acceptable salt thereof, characterized in that: Its structural formula is as follows: 。 4. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, characterized in that: The pharmaceutically acceptable salts are acid addition salts of the compounds.

5. The compound according to claim 4 or a pharmaceutically acceptable salt thereof, characterized in that: The acid used for salt formation is an inorganic acid or an organic acid, wherein the inorganic acid includes hydrochloric acid, sulfuric acid or phosphoric acid; the organic acid includes acetic acid, trichloroacetic acid, propionic acid, butyric acid, maleic acid, p-toluenesulfonic acid, malic acid, malonic acid, cinnamic acid, citric acid, fumaric acid, camphoric acid, digluconic acid, aspartic acid, tartaric acid or methanesulfonic acid.

6. A pharmaceutical composition, characterized in that It comprises the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt, enantiomer thereof, and a pharmaceutically acceptable carrier.

7. Use of the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 6 in the preparation of a PRMT7 target inhibitor drug.

8. Use of the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 6 in the preparation of a medicament for preventing and / or treating cancer or tumor-related diseases.

9. The use according to claim 8, characterized in that: The cancer or tumor-related disease includes prostate cancer, colorectal cancer, ovarian cancer, leukemia, lung cancer, liver cancer, breast cancer, esophageal cancer, glioma or pancreatic cancer.

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