Novel ezh2 inhibitor compounds and methods of making and using the same

Abstract

The present application relates to novel EZH2 inhibitor compounds and preparation method and application thereof, these compounds have structure formula shown in general formula (I) or (II) or (III), as an antitumor drug, have significant EZH2 histone methyltransferase inhibitory activity. The present application also provides the preparation method of the compound, and the pharmaceutical composition and use containing them. The novel EZH2 inhibitor compound obtained by the present application has more excellent antitumor activity and safety, can be used in the disease mediated by EZH2, PRC2, EZH2 / PRC2, especially as an antitumor agent has great value.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and particularly relates to novel EZH2 inhibitor compounds that inhibit tumor cell growth and exert anti-tumor effects, as well as their pharmaceutically acceptable salts, preparation methods, and pharmaceutical uses. It is a compound with therapeutic effects on EZH2, PRC2, and EZH2 / PRC2-mediated diseases. Background Technology

[0002] Cancer remains a significant public health challenge worldwide, with cancer incidence and mortality rates continuing to rise in my country. Epigenetics, a concept corresponding to genetics, primarily studies how regulatory mechanisms influence gene expression without altering the DNA sequence. These mechanisms include DNA methylation, histone post-translational modifications, and chromatin remodeling. Increasing research indicates a close correlation between cancer development and abnormal epigenetic expression. Polycomb group (PcG) proteins are important epigenetic regulatory molecules. PRC2 is a methyltransferase composed of embryonic ectoderm developmental protein (EED), Zeste 12 homolog repressor (SUZ 12), and Zeste homolog 1 / 2 enhancer (EZH1 / 2). These components collectively constitute PRC2, playing a crucial role in organisms. Zeste homolog 2 (EZH2) is an important catalytic subunit of the protein repressor complex PRC2. It can trimethylate the 27th lysine residue of histone H3 (H3K27me3) to promote transcriptional silencing and inhibit the expression of target genes. Most of these target genes have the functions of inhibiting cell proliferation and promoting cell differentiation. Overexpression or excessive enhancement of EZH2 function will lead to excessive silencing of the expression of these genes, ultimately resulting in tumorigenesis.

[0003] EPZ-6438 was the first EZH2 inhibitor approved by the FDA, primarily used to treat epithelioid sarcoma and relapsed or refractory follicular lymphoma. To date, many small-molecule EZH2-specific inhibitors have entered clinical trials, but many have shown limited clinical efficacy, such as preclinical resistance and poor treatment outcomes. Improving the anticancer activity of EZH2 requires new technologies and methods. This invention will design and synthesize a novel EZH2 inhibitor using Tazemetostat as a control drug.

[0004] A novel substituted thiophene-pyrimidine compound was synthesized previously, and its in vitro antitumor activity was studied. This invention builds upon that foundation for further research. Summary of the Invention

[0005] The purpose of this invention is to provide a novel class of EZH2 inhibitor compounds and their use in drugs for EZH2, PRC2, and EZH2 / PRC2-mediated diseases. After in vitro activity screening, the compound exhibits significant antitumor activity.

[0006] This invention relates to novel EZH2 inhibitor compounds represented by formula (I), (II), or (III) and their pharmaceutically acceptable salts:

[0007]

[0008] in,

[0009] A is selected from primary amine compounds;

[0010] P is selected from aryl or heteroaryl;

[0011] R is selected from secondary amines or structures containing secondary amine substitutions.

[0012] Furthermore, the aforementioned novel EZH2 inhibitor compounds and their pharmaceutically acceptable salts,

[0013] A is selected from 3-aminomethyl-4,6-dimethylpyridin-2(1H)one;

[0014] P is selected from 2,3-dihydro-1H-indene;

[0015] R is selected from morpholino, dimethylamino, N-methylpiperazinyl, cis-dimorpholino, methoxypyridinyl, 1-(2-methoxyethyl)piperazinyl, acetylpiperazinyl, or 4-dimethylaminopiperidine.

[0016] Furthermore, the aforementioned novel EZH2 inhibitor compound has the following structural formula:

[0017]

[0018]

[0019] According to some common methods in the field to which this invention pertains, the novel substituted thiophene-pyrimidine compounds of general formula (I) or (II) or (III) of this invention can react with acids to form pharmaceutically acceptable salts. Pharmaceutically acceptable addition salts include addition salts of inorganic acids and organic acids, with salts that react with the following acids being particularly preferred: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, and benzoic acid.

[0020] A pharmaceutical composition comprising any of the above-mentioned novel EZH2 inhibitor compounds and their pharmaceutically acceptable salts as active ingredients in combination with a pharmaceutically acceptable carrier.

[0021] Preferably, in the above-described pharmaceutical composition, the pharmaceutically acceptable carrier is selected from one or more fillers, disintegrants, binders, and lubricants.

[0022] Preferably, the above-mentioned pharmaceutical composition is formulated into dosage forms such as tablets, capsules, granules, sprays, or injections.

[0023] The use of any of the novel EZH2 inhibitor compounds described above, their pharmaceutically acceptable salts, or any of the pharmaceutical compositions described above in the preparation of medicaments for treating EZH2, PRC2, or EZH2 / PRC2-mediated diseases.

[0024] The use of any of the novel EZH2 inhibitor compounds described above, their pharmaceutically acceptable salts, or any of the pharmaceutical compositions described above in the preparation of EZH2 as a molecular therapeutic target drug.

[0025] The use of any of the novel EZH2 inhibitor compounds described above, their pharmaceutically acceptable salts, or any of the pharmaceutical compositions described above in the preparation of antitumor drugs.

[0026] Preferably, the tumors are: liver cancer, nasopharyngeal carcinoma, colorectal cancer, melanoma, bladder cancer, esophageal cancer with leukemia, breast cancer, gastric cancer, prostate cancer, pancreatic cancer, lung cancer, ovarian cancer, epithelioid sarcoma, non-Hodgkin's lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, and follicular lymphoma.

[0027] The beneficial effects of this invention are: the novel EZH2 inhibitor compound and its pharmaceutically acceptable salt obtained by this invention possess excellent antitumor activity and safety. They can be used to treat neoplastic diseases such as epithelioid sarcoma, lymphoma, prostate cancer, and breast cancer, as well as other autoimmune diseases. They have a high anticancer spectrum and a broad therapeutic window, making them of great application value in the pharmaceutical field. Detailed Implementation

[0028] 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.

[0029] The following synthetic routes describe the preparation methods of the novel EZH2 inhibitor compounds of general formula (I), (II), or (III) of this invention.

[0030] All raw materials were prepared by methods well known to those skilled in the art of organic chemistry, or were commercially available, as described in the synthetic routes below. All final compounds of this invention were prepared by methods described in the synthetic routes below or by similar methods well known to those skilled in the art of organic chemistry. All variable factors used in the synthetic routes below are as defined below or as defined in the claims.

[0031] 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 ARX-600, and the mass spectrometry was performed using an Agilent 1100 LC / MSD; all reagents used were analytical grade or chemically pure.

[0032] The synthetic routes for compounds 1-25 of general formula (I), (II), or (III) according to the present invention are as follows:

[0033] 1) As shown in the synthetic route 1 of the target compound, 5-bromo-1-indanone was reduced by NaBH4 in methanol solution to obtain intermediate 2; intermediate 2 was reacted with sulfoxide in dichloromethane solution to obtain intermediate 3; intermediate 3 was reacted with R to obtain intermediate 4; intermediate 4 was coupled with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxacyclopentaborane) to obtain intermediate 5.

[0034]

[0035] Among them, (a)MeOH,NaBH4,RT; (b)SOCl2,DCM,RT; (c)K2CO3,KI,MeCN,60℃; (d)Diox,Pd(pph3)2Cl2,CH3COOK,100℃.

[0036] 2) As shown in the synthetic route 2 of the target compound, 2-methyl-3-nitrobenzoic acid was reacted with dibromohydantoin at 25°C to obtain intermediate 7; intermediate 7 was esterified to obtain intermediate 8; intermediate 8 was reduced by iron powder to obtain intermediate 9; intermediate 9 was reacted with cyclopropionyl chloride in acetonitrile solution to obtain intermediate 10; intermediate 10 was reacted with iodoethane in DMF solution to obtain intermediate 11; intermediate 11 was hydrolyzed in methanol aqueous solution to obtain intermediate 12; intermediate 12 was reacted with 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one to obtain intermediate 13; intermediate 13 was coupled with boric acid to obtain the target compound (compound 1, compound 3, compound 6, compound 9, compound 12, compound 14, compound 17, compound 20, compound 23, compound 24).

[0037]

[0038] Among them, (a) H2SO4, 25℃; (b) MeOH, H2SO4, 60℃; (c) NH4Cl, MeOH, H2O, 80℃; (d) MeCN, K2CO3, RT; (e) Cs2CO3, DMF, Diox, RT; (f) NaOH, MeOH, H2O, 70℃; (g) HATU, DMF, Et3N, 50℃; (h) Diox, Pd(pph3)2Cl2, H2O, CH3COOK, 100℃.

[0039] 3) As shown in the synthetic route 3 of the target compound, methyl 6-bromo-4-indazolecarboxylate was used as a starting material and reacted with 2-iodopropane in acetonitrile solution to obtain intermediate 16; intermediate 16 was hydrolyzed to obtain intermediate 17; intermediate 17 was reacted with 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one to obtain intermediate 18; intermediate 18 was coupled with boric acid to obtain the target compound (compound 2, compound 4, compound 7, compound 10, compound 13, compound 15, compound 18, compound 21, compound 25).

[0040]

[0041]

[0042] Among them, (a) Cs2CO3, MeCN, 90℃; (b) NaOH, MeOH, H2O, 70℃; (c) HATU, DMF, Et3N, 25℃; (d) Diox, Pd(pph3)2Cl2, H2O, CH3COOK, 100℃.

[0043] 4) As shown in the synthetic route 4 of the target compound, methyl 6-bromo-4-indolecarboxylate was used as a starting material and reacted with 2-iodopropane in DMF solution to obtain intermediate 21; intermediate 21 was hydrolyzed to obtain intermediate 22; intermediate 22 was reacted with 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one to obtain intermediate 23; intermediate 23 was coupled with boric acid to obtain the target compound (compound 5, compound 8, compound 11, compound 16, compound 19, compound 22).

[0044]

[0045] Among them, (a) NaH, DMF; (b) NaOH, MeOH, H2O, 70℃; (c) HATU, DMF, Et3N, 25℃; (d) Diox, Pd(pph3)2Cl2, H2O, CH3COOK, 100℃.

[0046] Example 1

[0047]

[0048] N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-(furan-2-yl)thiopheno[3,2-d]pyrimidin-4-carboxamide (chemical)

[0049] Compound 1)

[0050] (1) Synthesis of 5-bromo-2,3-dihydro-1H-inden-1-ol (intermediate 2)

[0051]

[0052] Intermediate 1 (10 g, 0.05 mmol) was added to a 100 mL pear-shaped flask and dissolved in 50 mL of methanol solution. NaBH4 (1.93 g, 0.05 mmol) was added to a cold trap, and the reaction was carried out at room temperature. After 4 h, the reaction was complete. 9.93 g of white solid precipitated in cold water, with a yield of 98.82%.

[0053] (2) Synthesis of 5-bromo-1-chloro-2,3-dihydro-1H-indene (intermediate 3)

[0054]

[0055] Intermediate 2 (10 g, 0.05 mol) was added to a 250 mL flask and dissolved in 80 mL of dichloromethane. Thionyl chloride (11.27 g, 0.1 mol) was added dropwise into a cold trap, and the reaction was carried out at room temperature for 24 h until complete. Thionyl chloride was removed by rotary evaporation, and then 100 mL of dichloromethane was added to the resulting filtrate. The mixture was then washed twice with 50 mL of water to ensure removal of impurities. Next, it was washed twice with 30 mL of saturated NaCl solution each time to further purify the organic phase. After washing, the organic phase was dried with anhydrous sodium sulfate. Sodium sulfate was then removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry to obtain the desired crude product. Dry column chromatography yielded 8.54 g of the product, with a yield of 78.71%.

[0056] (3) Synthesis of 4-(5-bromo-2,3-dihydro-1H-inden-1-yl)morpholine (intermediate 4a)

[0057]

[0058] In a 50 mL three-necked flask, intermediate 3 (1 g, 0.04 mol), K₂CO₃ (3.01 g, 0.20 mol), and KI (0.72 g, 0.04 mol) were added and dissolved in 15 mL of acetonitrile solution. Stirring was started, and morpholine (0.76 g, 0.08 mol) was added. The reaction was carried out at 60 °C under nitrogen protection. After 24 h, the reaction was complete. Insoluble matter was removed by filtration. The filtrate was adjusted to pH 2, and the aqueous phase was retained by reverse extraction. The aqueous phase was then adjusted to pH 11, retaining the organic phase. 20 mL of dichloromethane was added to the organic phase. The mixture was washed twice with 10 mL of water, then twice with 10 mL of saturated NaCl solution each time. The organic phase was dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. Dry column chromatography was used to separate the crude product into 0.85 g of a brown solid, with a yield of 69.53%.

[0059] (4) Synthesis of 5-bromo-N,N-dimethyl-2,3-dihydro-1H-inden-1-amine (intermediate 4b)

[0060]

[0061] Intermediate 3 (1 g, 0.04 mol), K₂CO₃ (3.01 g, 0.20 mol), and KI (0.72 g, 0.04 mol) were added to a 50 mL three-necked flask and dissolved in 15 mL of acetonitrile solution. Stirring was started, and dimethylamine (0.39 g, 0.08 mol) was added. Using the reaction conditions and post-treatment method for intermediate 4a, 0.67 g of solid product was obtained, with a yield of 64.89%.

[0062] (5) Synthesis of 1-(5-bromo-2,3-dihydro-1H-inden-1-yl)-4-methylpiperazine (intermediate 4c)

[0063]

[0064] Intermediate 3 (1 g, 0.04 mol), K₂CO₃ (3.01 g, 0.20 mol), and KI (0.72 g, 0.04 mol) were added to a 50 mL three-necked flask and dissolved in 15 mL of acetonitrile solution. Stirring was started, and N-methylpiperazine (2.56 g, 0.08 mol) was added. Using the reaction conditions and post-treatment method for intermediate 4a, 0.83 g of solid was obtained, with a yield of 64.84%.

[0065] (6) Synthesis of (2S,6R)-4-(5-bromo-2,3-dihydro-1H-inden-1-yl)-2,6-dimethylmorpholine (intermediate 4d)

[0066] become

[0067]

[0068] Intermediate 3 (1 g, 0.04 mol), K₂CO₃ (3.01 g, 0.20 mol), and KI (0.72 g, 0.04 mol) were added to a 50 mL three-necked flask and dissolved in 15 mL of acetonitrile solution. Stirring was started, and cis-dimethylmorpholine (2.7 g, 0.08 mol) was added. Using the reaction conditions and post-treatment method of intermediate 4a, 0.77 g of solid product was obtained, with a yield of 57.21%.

[0069] (7) Synthesis of 1-(5-bromo-2,3-dihydro-1H-inden-1-yl)-4-methoxypiperidine (intermediate 4e)

[0070]

[0071] Intermediate 3 (1 g, 0.04 mol), K₂CO₃ (3.01 g, 0.20 mol), and KI (0.72 g, 0.04 mol) were added to a 50 mL three-necked flask and dissolved in 15 mL of acetonitrile solution. Stirring was started, and methoxypyridine (1.01 g, 0.08 mol) was added. Using the reaction conditions and post-treatment method for intermediate 4a, 0.73 g of solid was obtained, with a yield of 54.23%.

[0072] (8) Synthesis of 1-(5-bromo-2,3-dihydro-1H-inden-1-yl)-4-(2-methoxyethyl)piperazine (intermediate 4f).

[0073]

[0074] Intermediate 3 (1 g, 0.04 mol), K₂CO₃ (3.01 g, 0.20 mol), and KI (0.72 g, 0.04 mol) were added to a 50 mL three-necked flask and dissolved in 15 mL of acetonitrile solution. Stirring was started, and 1-(2-methoxyethyl)piperazine (1.26 g, 0.08 mol) was added. Using the reaction conditions and post-treatment methods for intermediate 4a, the product was purified to obtain 0.70 g of a brown solid, with a yield of 47.21%.

[0075] (9) Synthesis of 1-(1-(5-bromo-2,3-dihydro-1H-inden-1-yl)piperidin-4-yl)ethan-1-one (intermediate 4g)

[0076]

[0077] Intermediate 3 (1 g, 0.04 mol), K₂CO₃ (3.01 g, 0.2 mol), and KI (0.72 g, 0.04 mol) were added to a 50 mL three-necked flask and dissolved in 15 mL of acetonitrile solution. Stirring was started, and acetylpiperazine (1.10 g, 0.08 mol) was added. Using the reaction conditions and post-treatment methods for intermediate 4a, 1.06 g of a yellow solid was obtained after separation and purification, with a yield of 75.68%.

[0078] (10) Synthesis of 1-(5-bromo-2,3-dihydro-1H-inden-1-yl)-N,N-dimethylpiperidin-4-amine (intermediate 4h)

[0079] become

[0080]

[0081] Intermediate 3 (1 g, 0.04 mol), K₂CO₃ (3.01 g, 0.2 mol), and KI (0.72 g, 0.04 mol) were added to a 50 mL three-necked flask and dissolved in 15 mL of acetonitrile solution. Stirring was started, and 4-dimethylaminopiperidine (1.21 g, 0.08 mol) was added. Using the reaction conditions and post-treatment method of intermediate 4a, 0.87 g of a brown solid was obtained, with a yield of 61.71%.

[0082] (11) Synthesis of 1-(5-bromo-2,3-dihydro-1H-inden-1-yl)-4-methylpiperidine (intermediate 4i)

[0083]

[0084] Intermediate 3 (1 g, 0.04 mol), K₂CO₃ (3.01 g, 0.2 mol), and KI (0.72 g, 0.04 mol) were added to a 50 mL three-necked flask and dissolved in 15 mL of acetonitrile solution. Stirring was started, and 4-methylpiperidine (0.86 g, 0.08 mol) was added. Using the reaction conditions and post-treatment method for intermediate 4a, 0.73 g of solid was obtained, with a yield of 54.76%.

[0085] (12)N1-(5-bromo-2,3-dihydro-1H-inden-1-yl)-N 1 N 2 N 2 Synthesis of 1,2-trimethylethane-1,2-diamine (intermediate 4j)

[0086]

[0087] Add intermediate 3 (1g, 0.04mol), K2CO3 (3.01g, 0.2mol), and KI (0.72g, 0.04mol) to a 50mL three-necked flask, dissolve in 15mL of acetonitrile solution, start stirring, and add N2O. 1 N 1 N 2 -Triethylethane-1,2-diamine (0.89 g, 0.08 mol). Using the reaction conditions and post-treatment methods of intermediate 4a, 0.76 g of solid was obtained, with a yield of 53.21%.

[0088] (13)4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-

[0089] Synthesis of yl)morpholine (intermediate 5a)

[0090]

[0091] In a 50 mL three-necked flask, intermediate 4a (500 mg, 1.8 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxane) (678 mg, 2.7 mmol), and potassium acetate (261.18 mg, 2.7 mmol) were added and dissolved in 15 mL of dioxane solution. The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (62.41 mg, 0.09 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection for 12 h until the reaction was complete. The insoluble matter was removed by filtration, and 20 mL of dichloromethane was added to the organic phase. The mixture was washed twice with 10 mL of water, and then twice with 10 mL of saturated NaCl solution each time. The organic phase was dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. Dry column chromatography was used to separate and purify the solid to obtain 423 mg of solid, with a yield of 66.60%.

[0092] (14)N,N-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-

[0093] Synthesis of amine (intermediate 5b)

[0094]

[0095] Intermediate 4b (500 mg, 2.01 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxane-pentaborane) (797 mg, 3.13 mmol), and potassium acetate (307.18 mg, 3.13 mmol) were added to a 50 mL three-necked flask and dissolved in 15 mL of dioxane solution. The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (73.44 mg, 0.10 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment method of intermediate 5a, 387 mg of a yellow solid was obtained, with a yield of 76.14%.

[0096] (15)1-methyl-4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-

[0097] Synthesis of yl)piperazine (intermediate 5c)

[0098]

[0099] Intermediate 4c (500 mg, 1.70 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxane, 2.55 mmol), and potassium acetate (250.28 mg, 2.55 mmol) were added to a 50 mL three-necked flask and dissolved in 15 mL of dioxane solution. The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (62.4 mg, 0.08 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment method of intermediate 5a, 521 mg of a pale yellow solid was obtained, with a yield of 84.75%.

[0100] (16)(2S,6R)-2,6-dimethyl-4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-

[0101] Synthesis of inden-1-yl)morpholine (intermediate 5d)

[0102]

[0103] Intermediate 4d (500 mg, 1.62 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxane-pentaborane) (616 mg, 2.43 mmol), and potassium acetate (237.76 mg, 2.43 mmol) were added to a 50 mL three-necked flask and dissolved in 15 mL of dioxane solution. The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (56.78 mg, 0.08 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment method of intermediate 5a, 421 mg of a pale yellow solid was obtained, with a yield of 66.54%.

[0104] (17)4-methoxy-1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-

[0105] Synthesis of yl)piperidine (intermediate 5e)

[0106]

[0107] Intermediate 4e (500 mg, 1.62 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxane-pentaborane) (624 mg, 2.43 mmol), and potassium acetate (237.46 mg, 2.43 mmol) were added to a 50 mL three-necked flask and dissolved in 15 mL of dioxane solution. The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (56.79 mg, 0.08 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment method of intermediate 5a, 497 mg of solid product was obtained, with a yield of 78.62%.

[0108] (18)1-(2-methoxyethyl)-4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-

[0109] Synthesis of inden-1-yl)piperazine (intermediate 5f)

[0110]

[0111] Intermediate 4f (500 mg, 1.48 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxane, 2.22 mmol), and potassium acetate (217.36 mg, 2.22 mmol) were added to a 50 mL three-necked flask and dissolved in 15 mL of dioxane solution. The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (51.9 mg, 0.07 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment method of intermediate 5a, 468 mg of solid was obtained, with a yield of 68.42%.

[0112] (19) Synthesis of 1-(1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-yl)piperidin-4-yl)ethan-1-one (intermediate 5g)

[0113]

[0114] In a 50 mL three-necked flask, 4 g (500 mg, 1.55 mmol) of intermediate, 593 mg (2.32 mmol) of 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxane), and 228 mg (2.32 mmol) of potassium acetate were added and dissolved in 15 mL of dioxane solution. The mixture was bubbled under nitrogen for 30 min, and then 55 mg (0.08 mmol) of Pd(pph3)2Cl2 was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment method of intermediate 5a, 466 mg of a pale yellow solid was obtained, with a yield of 71.25%.

[0115] (20)N,N-dimethyl-1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-

[0116] Synthesis of yl)piperidin-4-amine (intermediate 5h)

[0117]

[0118] In a 50 mL three-necked flask, intermediate 4h (500 mg, 1.55 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxane) (591 mg, 2.32 mmol), and potassium acetate (228 mg, 2.32 mmol) were added and dissolved in 15 mL of dioxane solution. The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (54 mg, 0.08 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment method of intermediate 5a, 534 mg of solid was obtained, with a yield of 81.47%.

[0119] (21)4-methyl-1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-

[0120] Synthesis of yl)piperidine (intermediate 5i)

[0121] In a 50 mL three-necked flask, intermediate 4i (500 mg, 1.55 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxane-2,3,2-dioxane-2,3-2-borane) (647 mg, 2.32 mmol), and potassium acetate (250 mg, 2.32 mmol) were added and dissolved in 15 mL of dioxane solution. The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (59.28 mg, 0.08 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment method of intermediate 5a, 489 mg of solid was obtained, with a yield of 69.53%.

[0122] (22)N 1 N 1 N 2 -trimethyl-N2-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-

[0123] Synthesis of inden-1-yl)ethane-1,2-diamine (intermediate 5j)

[0124]

[0125] Intermediate 4j (500 mg, 1.55 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxane-pentaborane) (643 mg, 2.32 mmol), and potassium acetate (248 mg, 2.32 mmol) were added to a 50 mL three-necked flask and dissolved in 15 mL of dioxane solution. The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (59.28 mg, 0.08 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment method of intermediate 5a, 463 mg of a pale yellow solid was obtained, with a yield of 73.46%.

[0126] (23) Synthesis of 5-bromo-2-methyl-3-nitrobenzoic acid (intermediate 7)

[0127]

[0128] 2-Methyl-3-nitrobenzoic acid (30 g, 0.17 mol) was dissolved in 70 mL of concentrated sulfuric acid in a 1 L round-bottom flask. Stirring was started, and dibromohydantoin (8.71 g, 8.3 mmol) was added. The reaction was carried out at 25 °C for 5 h until complete. The reaction solution was then added to cold water, precipitating a large amount of white solid. After drying, 41.10 g of product was obtained, with a yield of 95.80%.

[0129] (24) Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate (intermediate 8)

[0130]

[0131] Intermediate 7 (30 g, 0.12 mol) was placed in a 1 L round-bottom flask and dissolved in 200 mL of methanol solution. 100 mL of concentrated sulfuric acid was slowly added dropwise into a cold trap, and the mixture was refluxed at 60 °C. After 6 h, the reaction was complete. The pH of the reaction solution was adjusted to about 9 with NaOH solution, and a large amount of pale yellow solid precipitated. After drying, 30.04 g of solid was obtained, with a yield of 94.67%.

[0132] (25) Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate (intermediate 9)

[0133]

[0134] Intermediate 8 (30 g, 0.11 mol) was added to a 1 L round-bottom flask and dissolved in 280 mL of methanol-water solution (3:1). Then, 14.7 g of ammonium chloride (29.24 g, 0.55 mol) was added, followed by iron powder (61.31 g, 1.1 mol). The reaction was carried out at 80 °C for approximately 9 hours until completion. The mixture was filtered using diatomaceous earth, and the filtrate was collected. Then, 100 mL of dichloromethane was added to the filtrate. The mixture was washed twice with 50 mL of water, and then twice with 30 mL of saturated NaCl solution each time. The organic phase was dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. 22.31 g of a white solid was separated by dry column chromatography, with a yield of 83.55%.

[0135] (26) Synthesis of methyl 5-bromo-3-(cyclopropanecarboxamido)-2-methylbenzoate (intermediate 10)

[0136]

[0137] Intermediate 9 (20 g, 0.08 mol) and K₂CO₃ (22.63 g, 0.16 mol) were added to a 500 mL round-bottom flask and dissolved in 100 mL of acetonitrile solution. Stirring was started, and cyclopropionyl chloride (8.52 g, 0.08 mol) was added dropwise. The reaction was allowed to proceed at room temperature for 6 hours until complete. The mixture was then filtered, and the filtrate was collected. Next, 100 mL of dichloromethane was added to the filtrate. The mixture was then washed twice with 50 mL of water to ensure the removal of impurities. Afterward, it was washed twice with saturated NaCl solution, 30 mL each time, to further purify the organic phase. After washing, the organic phase was dried with anhydrous sodium sulfate. The sodium sulfate was then removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry to obtain the desired crude product. Dry column chromatography was used to separate the product into 22.13 g of a white solid, with a yield of 87.79%.

[0138] (27) The synthesis of methyl5-bromo-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzoate (intermediate 11)

[0139] become

[0140]

[0141] Intermediate 10 (20 g, 0.06 mol) and Cs₂CO₃ (41.92 g, 0.12 mol) were added to a 250 mL three-necked flask and dissolved in 80 mL of LDM. Stirring was started, and iodoethane (20.06 g, 0.12 mol) was added dropwise. The reaction was allowed to proceed at room temperature for 3 hours until complete. The reaction solution was collected and filtered through filter paper. Next, 100 mL of dichloromethane was added to the filtrate. The mixture was then washed twice with 50 mL of water to ensure removal of impurities. Afterward, it was washed twice with 30 mL of saturated NaCl solution each time to further purify the organic phase. After washing, the organic phase was dried with anhydrous sodium sulfate. The sodium sulfate was then removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry to obtain the desired crude product. Dry column chromatography was used to purify the product, yielding 15.7 g of solid (74.40%).

[0142] (28) Synthesis of 5-bromo-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzoic acid (intermediate 12)

[0143]

[0144] Intermediate 11 (10 g, 0.03 mol) was added to a 250 mL round-bottom flask and dissolved in 140 mL of methanol-water solution (6:1). NaOH (2.94 g, 0.06 mol) was then added, and the mixture was refluxed at 70 °C for 2 hours until complete. Methanol was removed by rotary evaporation, and 20 mL of water was added. The pH was adjusted to approximately 5 using dilute hydrochloric acid. 100 mL of dichloromethane was added to the resulting filtrate. The mixture was then washed twice with 50 mL of water to ensure removal of impurities. Next, it was washed twice with 30 mL of saturated NaCl solution each time to further purify the organic phase. After washing, the organic phase was dried using anhydrous sodium sulfate. Sodium sulfate was then removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry to obtain the desired crude product. 8.75 g of a yellow solid was obtained, with a yield of 91.44%.

[0145] (29)5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(N-

[0146] Synthesis of ethylcyclopropanecarboxamido)-2-methylbenzamide (intermediate 13)

[0147]

[0148] Intermediate 12 (5 g, 0.02 mol) was dissolved in DMF, and triethylamine (2.31 g, 0.02 mol), 3-aminomethyl-4,6-dimethylpyridin-2(1H)one (4.67 g, 0.02 mol), and HATU (11.69 g, 0.02 mol) were added. The reaction was carried out at 25 °C. After 10 h, the reaction was not complete, so it was heated to 50 °C and the reaction was completed after 6 h. 50 mL of dichloromethane was added to the filtrate of the reaction solution. The mixture was then washed twice with 30 mL of water to ensure the removal of impurities. After that, it was washed twice with 20 mL of saturated NaCl solution each time to further purify the organic phase. After washing, the organic phase was dried with anhydrous sodium sulfate. Then, sodium sulfate was removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry to obtain the desired crude product. Slurry separation yielded 4.92 g of white solid, with a yield of 70.26%.

[0149] Synthesis of (30) methyl 6-bromo-1-isopropyl-1H-indazole-4-carboxylate (intermediate 16)

[0150]

[0151] In a 250 mL round-bottom flask, methyl 6-bromo-4-indazolecarboxylate (intermediate 15) (10 g, 0.04 mol) was added and dissolved in 100 mL of acetonitrile. Cs₂CO₃ (41.92 g, 0.08 mol) and 2-iodopropane (5 g, 0.06 mol) were then added. The mixture was heated to 90 °C and refluxed for approximately 2 h. Methanol was removed by rotary evaporation, and 100 mL of dichloromethane was added. The mixture was then washed twice with 50 mL of water to ensure the removal of impurities. Afterward, it was washed twice with 30 mL of saturated NaCl solution each time to further purify the organic phase. Following washing, the organic phase was dried using anhydrous sodium sulfate. Sodium sulfate was then removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry, yielding 10.71 g of a yellow solid, with a yield of 92.26%.

[0152] (31) Synthesis of 6-bromo-1-isopropyl-1H-indazole-4-carboxylic acid (intermediate 17)

[0153]

[0154] Intermediate 16 (10 g, 0.03 mol) was dissolved in 140 mL of methanol-water solution (6:1) in a 250 mL round-bottom flask. NaOH (2.74 g, 0.06 mol) was then added, and the mixture was refluxed at 70 °C for 2 h until complete. Methanol was removed by rotary evaporation, and 100 mL of dichloromethane was added to the resulting filtrate. The mixture was then washed twice with 50 mL of water to ensure removal of impurities. Next, it was washed twice with 30 mL of saturated NaCl solution each time to further purify the organic phase. After washing, the organic phase was dried with anhydrous sodium sulfate. Sodium sulfate was then removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry, yielding 8.58 g of a yellow solid, with a yield of 90.01%.

[0155] (32)6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-

[0156] Synthesis of indazole-4-carboxamide (intermediate 18)

[0157]

[0158] Intermediate 17 (5 g, 0.02 mol) was dissolved in 40 mL of LMF. Triethylamine (2.68 g, 0.03 mol), 3-aminomethyl-4,6-dimethylpyridin-2(1H)one (5.39 g, 0.04 mol), and HATU (13.48 g, 0.04 mol) were added. The reaction was carried out at 25 °C. After 12 h, the reaction was found to be incomplete, so the reaction was heated to 50 °C and completed after 8 h. 100 mL of dichloromethane was added. The mixture was then washed twice with 50 mL of water to ensure removal of impurities. Next, it was washed twice with saturated NaCl solution, 30 mL each time, to further purify the organic phase. After washing, the organic phase was dried with anhydrous sodium sulfate. Sodium sulfate was then removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry to obtain the desired crude product. Slurry separation yielded 4.92 g of a white solid, with a yield of 70.76%.

[0159] (33) Synthesis of methyl 6-bromo-1-isopropyl-1H-indole-4-carboxylate (intermediate 21)

[0160]

[0161] 10 g (0.04 mmol) of methyl 6-bromo-4-indolecarboxylate (intermediate 20) was added to a 250 mL round-bottom flask and dissolved in 100 mL of anhydrous DMF. NaH (1.42 g, 0.08 mmol) was added with stirring at 0 °C, and the mixture was stirred for 30 min. Iodopropane (14.23 g, 2 mmol) was then added. The reaction was allowed to proceed at room temperature for approximately 5 h until complete. 100 mL of dichloromethane was added to the resulting filtrate. The mixture was then washed twice with 50 mL of water to ensure the removal of impurities. Next, it was washed twice with 30 mL of saturated NaCl solution each time to further purify the organic phase. After washing, the organic phase was dried with anhydrous sodium sulfate. The sodium sulfate was then removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry, yielding 10.41 g of a yellow solid, with a yield of 89.23%.

[0162] (34) Synthesis of 6-bromo-1-isopropyl-1H-indole-4-carboxylic acid (intermediate 22)

[0163]

[0164] Intermediate 21 (10 g, 0.03 mol) was dissolved in 140 mL of methanol-water solution (6:1) in a 250 mL round-bottom flask. NaOH (2.71 g, 0.06 mol) was then added, and the mixture was refluxed at 70 °C for 2 hours until complete. Methanol was removed by rotary evaporation, and 20 mL of water was added. The pH was adjusted to approximately 5 with dilute hydrochloric acid, and 100 mL of dichloromethane was added to the resulting filtrate. The mixture was then washed twice with 50 mL of water to ensure removal of impurities. Next, it was washed twice with 30 mL of saturated NaCl solution each time to further purify the organic phase. After washing, the organic phase was dried with anhydrous sodium sulfate. Sodium sulfate was then removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry, yielding 8.87 g of a yellow solid, with a yield of 93.41%.

[0165] (35) Synthesis of 6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-indole-4-carboxamide (intermediate 23)

[0166]

[0167] Intermediate 22 (5 g, 0.02 mol) was dissolved in DMF, and triethylamine (5.39 g, 0.03 mol), 3-aminomethyl-4,6-dimethylpyridin-2(1H)one (8.11 g, 0.04 mol), and HATU (27.05 g, 0.04 mol) were added. The reaction was carried out at 25 °C for 13 h until complete. 50 mL of dichloromethane was added. The mixture was then washed twice with 30 mL of water to ensure removal of impurities. Next, it was washed twice with 20 mL of saturated NaCl solution each time to further purify the organic phase. After washing, the organic phase was dried with anhydrous sodium sulfate. Sodium sulfate was then removed by filtration. Finally, the filtrate was evaporated under reduced pressure until completely dry to obtain the desired crude product. Slurrying yielded 4.92 g of a white solid, with a yield of 70.06%.

[0168] (36)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(N-

[0169] ethylcyclopropanecarboxamido)-2-methyl-5-(1-morpholino-2,3-dihydro-1H-inden-5-

[0170] Synthesis of yl)benzamide (compound 1)

[0171] Intermediate 13 (100 mg, 0.22 mmol), intermediate 5a (143 mg, 0.44 mmol), and potassium acetate (32 mg, 0.33 mmol) were added to a 50 mL three-necked flask and dissolved completely in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. The reaction was completed after 36 h. Insoluble matter was removed by filtration. 20 mL of dichloromethane was added to the organic phase. The mixture was washed twice with 10 mL of water, and then twice with 10 mL of saturated NaCl solution each time. The organic phase was dried over anhydrous sodium sulfate, and the sodium sulfate was removed by filtration. The filtrate was evaporated to dryness under reduced pressure to obtain the crude product. Dry column chromatography was used to separate and purify the product, yielding 15 mg of a white solid (11.82%).

[0172] MS (ESI) m / z (%): 583.30 [M+H] + 605.30[M+Na] + . 1H NMR(600MHz,Chloroform-d)δ11.96(s,1H),7.91(s,1H),7.80-7.66(m,4H),7.61(s ,2H),6.27(s,1H),4.91(s,2H),4.66(s,1H),4.42(dt,J=14.0,7.0Hz,1H),4.16-3.9 9(m,4H),3.67(dt,J=11.5,5.8Hz,1H),3.37-3.08(m,2H),2.88(d,J=26.2Hz,4H),2. 70(d,J=58.2Hz,6H),2.50(s,5H),2.07(s,2H),1.77(s,4H),1.57(d,J=37.8Hz,3H).

[0173] (37)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-morpholino-2,3-

[0174] Synthesis of dihydro-1H-inden-5-yl)-1H-indazole-4-carboxamide (compound 2)

[0175]

[0176] Intermediate 18 (100 mg, 0.24 mmol), intermediate 5a (158 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 34 mg of a white solid was obtained, with a yield of 26.53%.

[0177] MS (ESI) m / z (%): 539.30 [M] + . 1H NMR(600MHz,Chloroform-d)δ12.03(s,1H),8.53(s,1H),7.95(d,J=38.5Hz,2H),7.65(s,1H),7.38(d,J=6.9Hz,3H),7.26(s,1H),5.90(s,1H ),4.80(s,1H),4.62(s,2H),4.30(s,1H),3.72(s,4H),2.88(d,J=68.0 Hz, 2H), 2.54 (d, J = 19.4Hz, 4H), 2.42 (s, 2H), 2.08 (s, 5H), 1.64 (s, 6H).

[0178] (38)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(1-(dimethylamino)-2,3-

[0179] dihydro-1H-inden-5-yl)-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzamide(Compound

[0180] 3) Synthesis

[0181]

[0182] Intermediate 13 (100 mg, 0.22 mmol), intermediate 5b (125 mg, 0.44 mmol), and potassium acetate (35 mg, 0.33 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 22 mg of a white solid was obtained, with a yield of 18.96%.

[0183] MS (ESI) m / z (%): 540.30 [M] + 541.30 [M+H] + . 1H NMR (600MHz, DMSO-d6) δ11.48(s,1H),8.37(s,1H),7.63(d,J=39.0Hz,2H),5.87(s,1H),4.32(s,2H),3.80(s,1H), 3.48(s,1H),2.50(s,10H),2.15(d,J=50.9Hz,9H),1.23(s,4H),1.03(s,3H),0.76(s,3H),0.55(d,J=41.2Hz,2H).

[0184] (39)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(dimethylamino)-2,3-

[0185] Synthesis of dihydro-1H-inden-5-yl)-1-isopropyl-1H-indazole-4-carboxamide (compound 4)

[0186]

[0187] Intermediate 18 (100 mg, 0.24 mmol), intermediate 5b (137 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 33 mg of a white solid was obtained, with a yield of 27.86%.

[0188] MS (ESI) m / z (%): 498.20 [M+H] + 520.20[M+Na] + . 1 H NMR(400MHz,DMSO-d6)δ11.57(s,1H),8.66(s,1H),8.37(s,1H),8.13(s,1H),7.97-7.40(m,4H),7.31(s,1H),6.70(s,1H),5.89(s,1 H), 5.19 (s, 1H), 4.78 (s, 1H), 4.41 (d, J = 22.8Hz, 2H), 3.04 (d, J = 69.6Hz, 5H), 2.32 (s, 2H), 2.16 (d, J = 38.1Hz, 5H), 1.78-1.01 (m, 8H).

[0189] (40)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(dimethylamino)-2,3-

[0190] Synthesis of dihydro-1H-inden-5-yl)-1-isopropyl-1H-indole-4-carboxamide (compound 5)

[0191]

[0192] Intermediate 23 (100 mg, 0.24 mmol), intermediate 5b (125 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment method of intermediate compound 1, 29 mg of a white solid was obtained, with a yield of 24.56%.

[0193] MS (ESI) m / z (%): 497.40 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ11.57(s,1H),8.30(s,1H),7.93(d,J=11.8Hz,1H),7.80(s,1H),7.75-7.38(m,5H ), 6.88 (s, 1H), 5.84 (s, 1H), 4.78 (s, 1H), 4.30 (s, 2H), 2.13 (d, J = 17.8Hz, 9H), 1.38 (s, 8H), 1.23 (s, 4H).

[0194] (41)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(N-

[0195] ethylcyclopropanecarboxamido)-2-methyl-5-(1-(4-methylpiperazin-1-yl)-2,3-dihydro-1H-

[0196] Synthesis of inden-5-yl)benzamide (compound 6)

[0197]

[0198] Intermediate 13 (100 mg, 0.22 mmol), intermediate 5c (125 mg, 0.44 mmol), and potassium acetate (32 mg, 0.33 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 42 mg of a white solid was obtained, with a yield of 32.63%.

[0199] MS (ESI) m / z (%): 596.40 [M+H] + 618.3 [M+Na] + . 1 H NMR(400MHz,DMSO-d6)δ11.51(s,1H),8.40(s,1H),7.66-7.58(m,1H),7.56(s,1H),7.53(s, 2H),7.32(d,J=7.8Hz,1H),5.87(s,1H),4.29(t,J=7.0Hz,3H),3.87(dd,J=14.1,7.4Hz,1H) ,3.04-2.62(m,3H),2.45-2.30(m,6H),2.26-2.08(m,13H),2.02(dq,J=14.7,8.4,7.4Hz,3H ), 1.39 (s, 1H), 1.03 (d, J = 14.2Hz, 3H), 0.75 (d, J = 4.1Hz, 2H), 0.57 (dd, J = 20.2, 5.6Hz, 2H).

[0200] (42)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(4-methylpiperazin-1-yl)-2,3-

[0201] Synthesis of dihydro-1H-inden-5-yl)-1H-indazole-4-carboxamide (compound 7)

[0202]

[0203] Intermediate 18 (100 mg, 0.24 mmol), intermediate 5c (125 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 32 mg of a white solid was obtained, with a yield of 24.63%.

[0204] MS (ESI) m / z (%): 553.40 [M+H] + 575.40 [M+Na] + . 1 H NMR(400MHz,DMSO-d6)δ11.53(s,1H),8.64(s,2H),7.95(s,1H),7.85(s,1H),7. 67-7.58(m,2H),7.34(d,J=7.8Hz,1H),5.88(s,1H),4.89(p,J=6.6Hz,1H),4.42 -4.25(m,3H),2.89(ddt,J=46.8,15.3,6.8Hz,2H),2.47-2.35(m,5H),2.28-1.9 7(m,12H),1.58(s,5H),1.39(s,1H),1.25(d,J=20.0Hz,1H),1.00-0.76(m,1H).

[0205] (43)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(4-

[0206] Synthesis of methylpiperazin-1-yl)-2,3-dihydro-1H-inden-5-yl)-1H-indole-4-carboxamide (compound 8)

[0207]

[0208] Intermediate 23 (100 mg, 0.24 mmol), intermediate 5c (125 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 42 mg of a white solid was obtained, with a yield of 31.53%.

[0209] MS (ESI) m / z (%): 552.40 [M+H] + 574.30 [M+Na] + . 1 H NMR (400MHz, DMSO-d6) δ11.56(s,1H),8.31(t,J=5.0Hz,1H),7.88(s,1H),7.76-7.52(m,6H),7.32(d,J=7.8Hz,1H),6.86(d,J=3.1Hz,1H),5.88(s ,1H),5.05-4.83(m,1H),4.37(d,J=5.0Hz,2H),4.29(t,J=6.8Hz,1H),3. 03-2.61(m,4H),2.22(s,4H),2.15-2.00(m,11H),1.46(d,J=6.6Hz,7H).

[0210] (44)N-(3-(3-(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)propanoyl)-5-(1-((2S,6R)-2,6-

[0211] dimethylmorpholino)-2,3-dihydro-1H-inden-5-yl)-2-methylphenyl)-N-

[0212] Synthesis of ethylcyclopropanecarboxamide (compound 9)

[0213]

[0214] Intermediate 13 (100 mg, 0.22 mmol), intermediate 5d (155 mg, 0.44 mmol), and potassium acetate (32 mg, 0.33 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 42 mg of a white solid was obtained, with a yield of 31.84%.

[0215] MS (ESI) m / z (%): 611.40 [M+H] + 633.40 [M+Na] + . 1 H NMR (600MHz, DMSO-d6) δ11.48(s,1H),8.35(s,1H),7.59-7.49(m,4H),7.34(d,J=7.7Hz,1H),5. 86(s,1H),4.36-4.24(m,3H),3.86(s,1H),3.56(d,J=61.8Hz,2H),2.91(s,1H),2.81(s,1H),2. 73(s,1H),2.36(d,J=10.3Hz,1H),2.21(d,J=12.3Hz,6H),2.15(s,4H),2.07-2.01(m,2H),1.98 (s, 1H), 1.20 (d, J = 36.4Hz, 2H), 1.08-0.94 (m, 10H), 0.80-0.69 (m, 2H), 0.56 (d, J = 33.6Hz, 2H).

[0216] (45)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-((2S,6R)-2,6-

[0217] dimethylmorpholino)-2,3-dihydro-1H-inden-5-yl)-1-isopropyl-1H-indazole-4-

[0218] Synthesis of carboxamide (compound 10)

[0219]

[0220] Intermediate 18 (100 mg, 0.24 mmol), intermediate 5d (171 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 21 mg of a white solid was obtained, with a yield of 15.63%.

[0221] MS (ESI) m / z (%): 568.40 [M+H] + 590.40 [M+Na] + . 1 H NMR (400MHz, DMSO-d6) δ11.57(s,1H),8.64(s,2H),8.10-7.81(m,2H),7.68(d,J=30.2Hz,2H),7.36(s,1H),5.88(s,1H),4.90(s,1H),4.34(d,J =24.4Hz,3H),3.17-2.60(m,4H),2.39(s,1H),2.30-2.00(m,7H),1.88( d,J=7.7Hz,1H),1.53(d,J=30.6Hz,6H),1.22(s,2H),1.13-0.66(m,7H).

[0222] (46)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-((2S,6R)-2,6-

[0223] Synthesis of dimethylmorpholino)-2,3-dihydro-1H-inden-5-yl)-1-isopropyl-1H-indole-4-carboxamide (h compound 11)

[0224]

[0225] Intermediate 23 (100 mg, 0.24 mmol), intermediate 5d (125 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 27 mg of a white solid was obtained, with a yield of 19.63%.

[0226] MS (ESI) m / z (%): 567.40 [M+H] + 589.40 [M+Na] + . 1 H NMR (400MHz, DMSO-d6) δ11.58(s,1H),8.28(s,1H),8.02(d,J=36.3Hz,1H),7.70(d,J=69.8Hz,2H),6.83(s,1H),5.89(s,1H),5.37(d,J=5 3.0Hz, 1H), 4.97 (s, 1H), 4.52 (d, J = 104.4Hz, 6H), 2.73 (s, 1H), 2.18 (d, J = 56.2Hz, 10H), 1.36 (d, J = 105.8Hz, 13H), 0.94 (d, J = 79.8Hz, 3H).

[0227] (47)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(N-

[0228] ethylcyclopropanecarboxamido)-5-(1-(4-methoxypiperidin-1-yl)-2,3-dihydro-1H-inden-5-

[0229] Synthesis of yl)-2-methylbenzamide (compound 12)

[0230]

[0231] Intermediate 13 (100 mg, 0.22 mmol), intermediate 5e (155 mg, 0.44 mmol), and potassium acetate (32 mg, 0.33 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 21 mg of a white solid was obtained, with a yield of 15.69%.

[0232] MS (ESI) m / z (%): 611.40 [M+H] + 633.40 [M+Na] + . 1 H NMR (400MHz, DMSO-d6) δ11.56(s,1H),8.65(s,1H),8.36(s,1H),8.07(s,1H),7.84(s,1H),7.67(d,J=19.4Hz,2H),7.36(s,1H ),5.88(s,1H),5.15(s,1H),5.00(s,1H),4.36(d,J=14.4Hz,4H),3.20(s,4H),2.34-1.94(m,17H),1.82(s,3H),1.49(s,8H).

[0233] (48)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(4-

[0234] methoxypiperidin-1-yl)-2,3-dihydro-1H-inden-5-yl)-1H-indazole-4-carboxamide(compound

[0235] 13) Synthesis

[0236]

[0237] Intermediate 18 (100 mg, 0.24 mmol), intermediate 5e (171 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 25 mg of a white solid was obtained, with a yield of 18.64%.

[0238] MS (ESI) m / z (%): 568.40 [M+H] + 590.30 [M+Na] + . 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),8.39(s,1H),7.55(d,J=11.2Hz,4H),5.8 7(s,1H),4.30(s,3H),3.86(d,J=7.1Hz,1H),3.20(s,3H),3.00-2.78(m,2H),2 .20(d,J=8.8Hz,7H),2.10(s,5H),1.84(s,2H),1.39(s,2H),1.20(d,J=24.7Hz ,2H),1.03(t,J=7.1Hz,3H),0.92(s,1H),0.76(s,2H),0.58(d,J=11.4Hz,2H).

[0239] (49)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(N-

[0240] ethylcyclopropanecarboxamido)-5-(1-(4-(2-methoxyethyl)piperazin-1-yl)-2,3-dihydro-1H-

[0241] Synthesis of inden-5-yl)-2-methylbenzamide (compound 14)

[0242]

[0243] Intermediate 13 (100 mg, 0.22 mmol), intermediate 5f (168 mg, 0.44 mmol), and potassium acetate (32 mg, 0.33 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 33 mg of a white solid was obtained, with a yield of 23.67%.

[0244] MS (ESI) m / z (%): 640.40 [M+H] + 662.40 [M+Na] + . 1 H NMR (400MHz, DMSO-d6) δ11.51(s,1H),8.39(s,1H),7.95(s,1H),7.55(d,J=12.7H z,4H),7.33(d,J=7.4Hz,1H),5.87(s,1H),4.11(s,3H),3.88(s,1H),3.21(s,4H) ,2.87(d,J=31.7Hz,3H),2.40(s,6H),2.20(d,J=9.0Hz,6H),2.08(d,J=20.8Hz,6 H), 1.59-1.08 (m, 4H), 1.03 (t, J = 7.0Hz, 2H), 0.75 (s, 3H), 0.58 (d, J = 13.8Hz, 2H).

[0245] (50)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(4-(2-

[0246] Synthesis of methoxyethyl)piperazin-1-yl)-2,3-dihydro-1H-inden-5-yl)-1H-indazole-4-carboxamide (compound 15)

[0247]

[0248] Intermediate 18 (100 mg, 0.24 mmol), intermediate 5f (185.56 mg, 0.48 mmol), and potassium acetate (35.36 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8.06 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 24 mg of a white solid was obtained, with a yield of 16.53%.

[0249] MS (ESI) m / z (%): 597.40 [M+H]+. 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),8.64(s,1H),8.37(s,1H),8.07(s,1H),7.85(s,1H),7.67(d,J=23.0Hz,3H),7.34 (s,1H),5.88(s,1H),4.34(d,J=27.8Hz,5H),3.21(s,7H),2.90(d,J=41.8Hz,4H),2.16(d,J=38.2Hz,12H),1.23(s,6H).

[0250] (51)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(4-(2-

[0251] Synthesis of methoxyethyl)piperazin-1-yl)-2,3-dihydro-1H-inden-5-yl)-1H-indole-4-carboxamide (compound 16)

[0252]

[0253] Intermediate 23 (100 mg, 0.24 mmol), intermediate 5f (186 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 25 mg of a white solid was obtained, with a yield of 17.65%.

[0254] MS (ESI) m / z (%): 596.40 [M+H]+ . 1 H NMR (400MHz, DMSO-d6) δ11.56(s,1H),8.30(s,1H),8.11-7.89(m,1H),7.87-7.72(m,1H),7.72-7. 46(m,2H),6.87(d,J=2.8Hz,1H),5.85(s,1H),5.41(d,J=75.7Hz,1H),4.96(d,J=36.6Hz,1H),4.8 4-4.62(m,1H),4.54(s,1H),4.30(d,J=4.4Hz,1H),3.56(d,J=44.0Hz,7H),3.25-2.94(m,2H),2.6 6(s,1H),2.20-1.93(m,4H),1.80-1.29(m,7H),1.23(s,7H),1.09-0.99(m,2H),0.94-0.69(m,3H).

[0255] (52)5-(1-(4-acetylpiperazin-1-yl)-2,3-dihydro-1H-inden-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-

[0256] dihydropyridin-3-yl)methyl)-3-(N-ethylcyclopropanecarboxamido)-2-

[0257] Synthesis of methylbenzamide (compound 17)

[0258]

[0259] Intermediate 13 (100 mg, 0.22 mmol), intermediate 5 g (160 mg, 0.44 mmol), and potassium acetate (32 mg, 0.33 mmol) were added to a 50 mL three-necked flask and dissolved completely in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 32 mg of a white solid was obtained, with a yield of 23.61%.

[0260] MS (ESI) m / z (%): 624.30 [M+H] + 646.30 [M+Na] + . 1H NMR (600MHz, DMSO-d6) δ11.48(s,1H),8.36(s,1H),7.57(s,1H),7.52(d,J=12.9Hz,3H),7.36(d,J=7. 7Hz,1H),5.87(s,1H),4.37-4.29(m,3H),3.92-3.83(m,1H),3.48(d,J=17.3Hz,2H),2.97-2.77(m,2H) ,2.43(s,1H),2.37(d,J=1.9Hz,1H),2.32(s,1H),2.21(d,J=11.7Hz,6H),2.11(s,3H),2.05(d,J=7.5H z, 2H), 1.97 (s, 3H), 1.39 (s, 4H), 1.03 (t, J = 6.9Hz, 4H), 0.76 (d, J = 3.8Hz, 2H), 0.56 (d, J = 20.4Hz, 2H).

[0261] (53)6-(1-(4-acetylpiperazin-1-yl)-2,3-dihydro-1H-inden-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-

[0262] Synthesis of dihydropyridin-3-yl)methyl)-1-isopropyl-1H-indazole-4-carboxamide (compound 18)

[0263] Intermediate 18 (100 mg, 0.24 mmol), intermediate 5 g (177 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 30 mg of a white solid was obtained, with a yield of 21.32%.

[0264] MS (ESI) m / z (%): 581.30 [M+H] + 603.30[M+Na] + . 1H NMR (600MHz, DMSO-d6) δ11.50 (s, 1H), 8.62 (d, J = 19.0Hz, 2H), 7.96 (s, 1H), 7.85 (s, 1H), 7. 68-7.58(m,2H),7.37(d,J=7.7Hz,1H),5.87(s,1H),4.90(dt,J=13.0,6.4Hz,1H),4.39-4. 34(m,3H),3.49(d,J=12.6Hz,1H),3.41(s,3H),2.90(ddt,J=66.6,15.6,7.2Hz,2H),2.21( s,3H),2.11(s,3H),2.05(q,J=6.9Hz,2H),1.97(s,3H),1.58(d,J=6.5Hz,7H),1.39(s,3H).

[0265] (54)6-(1-(4-acetylpiperazin-1-yl)-2,3-dihydro-1H-inden-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-

[0266] Synthesis of dihydropyridin-3-yl)methyl)-1-isopropyl-1H-indole-4-carboxamide (compound 19)

[0267]

[0268] Intermediate 23 (100 mg, 0.24 mmol), intermediate 5 g (125 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and dissolved completely in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 33 mg of a white solid was obtained, with a yield of 23.58%.

[0269] MS (ESI) m / z (%): 580.30 [M+H] + 602.20[M+Na] + . 1H NMR(600MHz,DMSO-d6)δ11.53(s,1H),8.28(s,1H),7.89(s,1H),7.78-7.50(m,5H),7.36(s,1H),6.87(s,1H),5.88 (s,1H),4.95(d,J=28.7Hz,2H),4.38(s,4H),2.91(d,J=64.9Hz,3H),2.23(s,3H),2.17-1.93(m,10H),1.47(s,8H).

[0270] (55)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(1-(4-

[0271] (dimethylamino)piperidin-1-yl)-2,3-dihydro-1H-inden-5-yl)-3-(N-

[0272] Synthesis of ethylcyclopropanecarboxamido)-2-methylbenzamide (compound 20)

[0273]

[0274] Intermediate 13 (100 mg, 0.22 mmol), intermediate 5h (178 mg, 0.44 mmol), and potassium acetate (35 mg, 0.33 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 44 mg of a white solid was obtained, with a yield of 32.56%.

[0275] MS (ESI) m / z (%): 624.30 [M+H] + 646.30 [M+Na] + . 1H NMR (400MHz, DMSO-d6) δ11.51(s,1H),8.39(t,J=4.9Hz,1H),7.52(d,J=22.0Hz,4H),7.32(d,J=7.8Hz,1H),5.87(s,1H) ,4.30(d,J=4.5Hz,3H),3.86(dt,J=15.4,8.2Hz,1H),3.50(s,1H),2.91(dt,J=13.5,6.1Hz,2H),2.80(dt,J=16.1,7.8Hz ,1H),2.26(d,J=11.2Hz,1H),2.23-2.16(m,12H),2.09(d,J=8.2Hz,4H),2.02(dt,J=14.6,7.3Hz,3H),1.71(dd,J=29.0, 11.4Hz, 2H), 1.42 (d, J = 22.5Hz, 2H), 1.25-1.12 (m, 2H), 1.03 (t, J = 7.1Hz, 3H), 0.75 (d, J = 4.1Hz, 2H), 0.63-0.50 (m, 2H).

[0276] (56)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(4-

[0277] (dimethylamino)piperidin-1-yl)-2,3-dihydro-1H-inden-5-yl)-1-isopropyl-1H-indazole-4-

[0278] Synthesis of carboxamide (compound 21)

[0279]

[0280] Intermediate 18 (100 mg, 0.24 mmol), intermediate 5h (125 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 50 mg of a white solid was obtained, with a yield of 39.46%.

[0281] MS (ESI) m / z (%): 581.30 [M+H] + 603.30[M+Na] +. 1 H NMR(400MHz,DMSO-d6)δ11.54(s,1H),8.64(s,2H),7.95(s,1H),7.85(s,1H),7.64-7.55(m ,2H),7.33(d,J=7.9Hz,1H),5.87(s,1H),4.90(p,J=6.6Hz,1H),4.37(d,J=4.5Hz,2H),4.3 1(t,J=6.8Hz,1H),2.96-2.80(m,3H),2.26(d,J=11.3Hz,1H),2.21(s,4H),2.16(s,7H),2. 11 (s, 4H), 2.02 (d, J = 7.0Hz, 3H), 1.57 (d, J = 6.5Hz, 6H), 1.49 (d, J = 6.5Hz, 1H), 1.39 (s, 3H).

[0282] (57)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(4-

[0283] (dimethylamino)piperidin-1-yl)-2,3-dihydro-1H-inden-5-yl)-1-isopropyl-1H-indole-4-

[0284] Synthesis of carboxamide (compound 22)

[0285]

[0286] Intermediate 23 (100 mg, 0.24 mmol), intermediate 5h (125 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 47 mg of the product was obtained, with a yield of 32.48%.

[0287] MS (ESI) m / z (%): 580.30 [M+H] + 602.30[M+Na] + . 1H NMR (400MHz, DMSO-d6) δ8.31(t,J=4.9Hz,1H),7.89(s,1H),7.69(s,1H),7.61(dd,J=11.3,8.2Hz ,3H),7.33(d,J=7.8Hz,1H),6.87(d,J=3.2Hz,1H),5.88(s,1H),4.93(q,J=6.6Hz,1H),4.37(d,J =5.1Hz,2H),4.32(d,J=6.8Hz,1H),2.99-2.87(m,3H),2.85(t,J=7.8Hz,1H),2.61-2.52(m,3H), 2.25(s,8H),2.22(s,3H),2.11(s,3H),2.04(t,J=7.1Hz,2H),1.47(d,J=6.7Hz,7H),1.39(s,2H).

[0288] (58)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(1-((2-

[0289] (dimethylamino)ethyl)(methyl)amino)-2,3-dihydro-1H-inden-5-yl)-3-(N-

[0290] Synthesis of ethylcyclopropanecarboxamido)-2-methylbenzamide (compound 23)

[0291]

[0292] Intermediate 13 (100 mg, 0.24 mmol), intermediate 5i (148 mg, 0.48 mmol), and potassium acetate (32 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 34 mg of the product was obtained, with a yield of 24.56%.

[0293] MS (ESI) m / z (%): 595.30 [M+H] + 617.30 [M+Na] + . 1H NMR (400MHz, DMSO-d6) δ11.54(s,1H),8.39(t,J=4.2Hz,1H),7.68-7.37(m,4H),7.31(d,J=7.7Hz, 1H),5.87(s,1H),4.29(s,3H),3.87(dd,J=13.4,6.7Hz,1H),3.65-3.46(m,1H),2.85(dd,J=36.9,6 .9Hz,3H),2.45(d,J=11.1Hz,1H),2.20(d,J=8.9Hz,6H),2.15-1.99(m,6H),1.89(s,1H),1.56(dd, J=33.7,11.4Hz,2H),1.40-1.13(m,4H),1.02(t,J=5.7Hz,5H),0.88(d,J=6.2Hz,3H),0.76(s,2H).

[0294] (59)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(1-((2-

[0295] (dimethylamino)ethyl)(methyl)amino)-2,3-dihydro-1H-inden-5-yl)-3-(N-

[0296] Synthesis of ethylcyclopropanecarboxamido)-2-methylbenzamide (compound 24)

[0297]

[0298] Intermediate 13 (100 mg, 0.24 mmol), intermediate 5j (149 mg, 0.48 mmol), and potassium acetate (32 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (7 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 49 mg of the product was obtained, with a yield of 29.65%.

[0299] MS (ESI) m / z (%): 598.30 [M+H] + 602.30[M+Na] + . 1H NMR (400MHz, DMSO-d6) δ11.56(s,1H),8.39(t,J=4.8Hz,1H),8.06(s,1H),7.60-7.46(m,3H),7.31(d,J=7 .8Hz,1H),5.86(s,1H),4.65(s,1H),4.40-4.28(m,2H),3.86(dd,J=13.6,7.0Hz,1H),3.50(s,3H),3.04- 2.73(m,2H),2.43(d,J=6.8Hz,1H),2.34(dd,J=11.7,6.6Hz,2H),2.20(d,J=9.1Hz,4H),2.12(d,J=14.2H z, 8H), 2.06-1.90 (m, 2H), 1.46 (s, 1H), 1.19 (d, J = 22.3Hz, 5H), 1.02 (t, J = 7.1Hz, 3H), 0.93-0.66 (m, 4H).

[0300] (60)N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-((2-

[0301] (dimethylamino)ethyl)(methyl)amino)-2,3-dihydro-1H-inden-5-yl)-1-isopropyl-1H-

[0302] Synthesis of indazole-4-carboxamide (compound 25)

[0303]

[0304] Intermediate 18 (100 mg, 0.24 mmol), intermediate 5J (165 mg, 0.48 mmol), and potassium acetate (35 mg, 0.36 mmol) were added to a 50 mL three-necked flask and thoroughly dissolved in 12 mL of dioxane aqueous solution (3:1). The mixture was bubbled under nitrogen for 30 min, and then Pd(pph3)2Cl2 (8 mg, 0.01 mmol) was added. The mixture was evacuated three times and reacted at 100 °C under nitrogen protection. Using the reaction conditions and post-treatment methods for the final product compound 1, 34 mg of the product was obtained, with a yield of 27.61%.

[0305] MS (ESI) m / z (%): 555.30 [M+H] + 572.20 [M+Na] + . 1H NMR (400MHz, DMSO-d6) δ11.57(s,1H),8.66(d,J=4.9Hz,1H),8.37(s,1H),8.08(d,J=9.0Hz,1H), 7.86(s,1H),7.68(dd,J=19.0,10.9Hz,2H),7.36(d,J=8.0Hz,1H),5.89(s,1H),5.17(p,J=6.4Hz, 1H),4.41(dd,J=11.5,5.9Hz,3H),3.00-2.81(m,2H),2.46(d,J=7.2Hz,2H),2.40-2.30(m,2H),2 .22(s,3H),2.17(s,3H),2.12(s,10H),1.67(s,1H),1.58(d,J=6.6Hz,1H),1.49(d,J=6.5Hz,5H).

[0306] Example 2: In vitro antitumor cell activity

[0307] Compounds containing substituted thiophene-pyrimidine structures were screened for their in vitro inhibitory activity against human diffuse large B lymphoma cells (WSU-DLCL-2) and human B lymphoma cells (SU-DHL-6).

[0308] 1) Plating Procedure: Select SU-DHL-6 cells in the logarithmic growth phase, centrifuge to remove the supernatant, and then resuspend the cells in 1 mL of culture medium. Next, take 100 μL of the cell suspension and mix it with 900 μL of culture medium for a 10-fold dilution, and accurately count the cells using a cell counting chamber. Then, seed 100 μL of the cell suspension into each well of a 96-well plate, at a cell density of 1 × 10⁶ cells per well. 4 Cells. After inoculation, place the 96-well plate in an incubator and maintain the culture environment for 24 hours.

[0309] 2) Sample Preparation Stage: Accurately weigh the compound to be tested and the Tazematostat drug as a positive control. First, completely dissolve the sample in 100 μL of DMSO, then add 50 μL of Tween 80 to enhance solubility, and adjust the total volume to 2 mL with RPMI-1640 basal medium. After thorough mixing, ensure the solution is clear and transparent; at this point, the stock solution concentration is 1000 μmol / L.

[0310] 3) Drug Addition Procedure: The drug was serially diluted 10-fold at concentrations of 100 μmol / L, 10 μmol / L, 1 μmol / L, 0.1 μmol / L, and 0.01 μmol / L, preparing a total of five different concentrations of drug solution. Then, 100 μL of the drug solution was added to each well of a 96-well plate. Due to mixing with the existing culture medium in the wells, the drug concentration was further diluted by two times, resulting in final test concentrations of 50 μmol / L, 5 μmol / L, 0.5 μmol / L, 0.05 μmol / L, and 0.005 μmol / L. After drug addition, the cells were cultured for another 48 hours for subsequent assays.

[0311] 4) Detection Procedure: Under light-protected conditions, 20 μL of 0.5% MTT solution was added to each well of a 96-well plate using a pipette, and the plate was then incubated at 37°C for 4 hours. Afterward, the plate was centrifuged at 1500 rpm for 5 minutes to remove the supernatant. Subsequently, 150 μL of DMSO solution was added to each well, and the plate was vibrated for 5 minutes using a microplate reader to ensure complete dissolution of the blue-purple MTT crystals. Finally, the optical density of each well was measured at a wavelength of 490 nm. After the measurements, the cell inhibition rate at each drug concentration was calculated based on the obtained data, and the half-maximal inhibitory concentration (IC50) of the drug was further determined.

[0312] Table 1 Results of the in vitro antitumor cell activity of the compounds

[0313]

Claims

1. An EZH2 inhibitor compound and its pharmaceutically acceptable salt, characterized in that, The EZH2 inhibitor compounds and their pharmaceutically acceptable salts have the general structural formula shown in (I), (II), or (III): Wherein, A is selected from 3-aminomethyl-4,6-dimethylpyridin-2(1H)one, with the structural formula shown in (Ⅳ): (Ⅳ); P is selected from 2,3-dihydro-1H-indene, with the structural formula shown in (Ⅴ): (V), where any of the three saturated methylene groups can be replaced by R; R is selected from morpholino, dimethylamino, N-methylpiperazinyl, cis-dimorpholino, 1-(2-methoxyethyl)piperazinyl, acetylpiperazinyl, or 4-dimethylaminopiperidinyl.

2. EZH2 inhibitor compounds and their pharmaceutically acceptable salts, characterized in that, The EZH2 inhibitor compound has the following structural formula:

3. A pharmaceutical composition, characterized in that, It is prepared by combining the EZH2 inhibitor compound of claim 1 or 2 and its pharmaceutically acceptable salt as the active ingredient with a pharmaceutically acceptable carrier.

4. The pharmaceutical composition according to claim 3, characterized in that, The pharmaceutically acceptable carrier is selected from one or more of fillers, disintegrants, binders, and lubricants.

5. The pharmaceutical composition according to claim 3, characterized in that, Formulating a pharmaceutical composition into a dosage form such as tablets, capsules, granules, sprays, or injections.

6. The use of the EZH2 inhibitor compound of any one of claims 1-2 and its pharmaceutically acceptable salt, or the pharmaceutical composition of any one of claims 3-5, in the preparation of a medicament for treating EZH2, PRC2, or EZH2 / PRC2-mediated diseases.

7. The use of the EZH2 inhibitor compound of any one of claims 1-2 and its pharmaceutically acceptable salt, or the pharmaceutical composition of any one of claims 3-5, in the preparation of EZH2 as a molecular therapeutic target drug.

8. The use of the EZH2 inhibitor compound of any one of claims 1-2 and its pharmaceutically acceptable salt, or the pharmaceutical composition of any one of claims 3-5, in the preparation of an antitumor drug.

9. The application according to claim 8, characterized in that, The tumors mentioned are: liver cancer, nasopharyngeal carcinoma, colorectal cancer, melanoma, bladder cancer, esophageal cancer with leukemia, breast cancer, stomach cancer, prostate cancer, pancreatic cancer, lung cancer, ovarian cancer, epithelioid sarcoma, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, and follicular lymphoma.

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