Novel compound containing biphenyl amide analogue structure and application thereof

Abstract

The invention discloses a novel compound containing a biphenyl amide analogue structure and application of the novel compound, and belongs to the technical field of medicines. The compounds have a structural formula as shown in a general formula (I) or (II), and have remarkable EZH2 histone methyltransferase inhibitory activity as an anti-tumor drug. The invention also provides a preparation method of the compounds, a pharmaceutical composition containing the compounds and application of the pharmaceutical composition. The obtained novel compound containing the biphenyl amide analogue structure has better anti-tumor activity and safety, can be applied to EZH2, PRC2 and EZH2 / PRC2 mediated diseases, and particularly has great value as an anti-tumor agent. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to novel compounds containing a biphenylamide analogue structure that inhibit the growth of tumor cells and exhibit anti-tumor effects, and pharmaceutically acceptable salts thereof, as well as preparation methods and pharmaceutical uses. It is a compound that has a therapeutic effect on diseases mediated by EZH2, PRC2, and EZH2 / PRC2. Background Art

[0002] The causes of cancer are extremely diverse, involving multiple fields such as genetics, cell biology, pathology, epigenetics, and treatment response. With the in-depth understanding of the cancer mechanism, the epigenetic mechanism has become an important driving factor for cancer to acquire hallmark capabilities. Epigenetics controls the transcription and post-transcriptional regulation of a large number of genes, which mediate various cellular processes and functions, including proliferation, differentiation, invasion, survival, growth, metabolism, and immune response. The development and progression of many pathological conditions, including cancer, trauma, and infectious and autoimmune diseases, may be driven by abnormal epigenetic modifications. EZH2 belongs to the core members of the polycomb gene family and plays a key role in epigenetic regulation, especially in gene silencing and cell fate determination. The polycomb gene family includes PRC1 and PRC2, which maintain the functions of gene repression and gene silencing, respectively. EZH2 is the enzymatic catalytic subunit of PRC2. PRC2 mainly mediates the trimethylation of lysine 9 / 27 (K9 / K27) of histone H3 through the SET domain of the EZH2 gene via its characteristic methyltransferase activity, thereby silencing the expression of a series of downstream target genes including tumor suppressor genes (such as cell differentiation, tumor metastasis, etc.). It has now been found that the overexpression of EZH2 is closely related to the occurrence and development of various cancers, and EZH2 has become a core target for epigenetic therapy.

[0003] Tazemetostat was developed by Epizyme and was approved for marketing by the US Food and Drug Administration (FDA) in January 2020. It is the first approved EZH2 inhibitor. This treatment regimen is applicable to the population of histologically confirmed unresectable locally advanced / metastatic epithelioid sarcoma, covering adult and adolescent patients aged ≥16 years. Especially for patients with epithelioid sarcoma lacking INI1 (SMARCB1). And for the treatment of adult patients with relapsed or refractory follicular lymphoma, its approval provides a new treatment option for patients with epithelioid sarcoma and follicular lymphoma, and it is an innovative targeted therapeutic drug. However, due to its serious adverse reactions such as anemia and thrombocytopenia, as well as problems such as clinical drug resistance and selectivity, the long-term safety and potential risk of secondary malignancies still need further study. Therefore, the development of novel inhibitors with novel structures, safety, and effectiveness is a key area in the research of anti-tumor drugs at home and abroad. Summary of the Invention

[0004] Based on summarizing the structure-activity relationship of EZH2 kinase inhibitors and analyzing the three-dimensional spatial structure of EZH2 protein, the present invention designed and synthesized a series of novel compounds containing a biphenylamide analog structure. The aim is to provide a class of novel compounds containing a biphenylamide analog structure and their uses in drugs for diseases mediated by EZH2, PRC2, and EZH2 / PRC2. Through in vitro activity screening, the compounds have significant anti-tumor activity.

[0005] The technical solution adopted by the present invention is: a novel compound containing a biphenylamide analog structure and its pharmaceutically acceptable salt, having a structural general formula as shown in (Ⅰ) or (Ⅱ):

[0006]

[0007] Among them,

[0008] A is selected from primary amine compounds;

[0009] B is selected from isopropyl or cyclopentyl;

[0010] X is selected from C or N atoms;

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

[0012] Furthermore, for the above-mentioned novel compound containing a biphenylamide analog structure and its pharmaceutically acceptable salt,

[0013] A is selected from 3-aminomethyl-4,6-dimethylpyridin-2(1H)-one, 3-aminopiperidine-2,6-dione, 2-(3-indolyl)ethylamine, or 2-(1H-imidazol-4-yl)ethylamine;

[0014] R is selected from (2S,6R)-2,6-dimethylmorpholine, 1-methylpiperazine, 4-methoxypiperidine, 4-methylpiperidine, 1-(2-methoxyethyl)piperazine, 1-(piperazin-1-yl)ethanone, 3-isopropoxyazetidine, morpholine, 1-ethylpiperazine, 1-methanesulfonylpiperazine, or dimethylamine.

[0015] Furthermore, the present invention preferably relates to a novel compound containing a biphenylamide analog structure and its pharmaceutically acceptable salt having the structure shown in general formula (Ⅰ) or (Ⅱ), specifically the following structures:

[0016]

[0017]

[0018]

[0019] A pharmaceutical composition is prepared by combining the novel compound containing a biphenylamide analog structure and its pharmaceutically acceptable salt as active ingredients with a pharmaceutically acceptable carrier.

[0020] Furthermore, for the above-mentioned pharmaceutical composition, the pharmaceutically acceptable carrier is selected from one or more of fillers, disintegrants, binders, and lubricants.

[0021] Furthermore, for the above-mentioned pharmaceutical composition, the pharmaceutical composition is made into dosage forms such as tablets, capsules, granules, sprays, or injections.

[0022] According to some common methods in the field to which the present invention pertains, the novel compound containing a biphenylamide analog structure represented by general formula (I) or (II) in the present invention can form pharmaceutically acceptable salts with acids. Pharmaceutically acceptable addition salts include inorganic acid and organic acid addition salts, and the salts formed with the following acids are 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, etc.

[0023] In addition, the present invention also includes prodrugs of derivatives of the present invention. The prodrugs of derivatives of the present invention are derivatives of general formula (I) or (II), which may themselves have weak activity or even no activity, but after administration, they are converted into the corresponding bioactive forms under physiological conditions (such as through metabolism, solvolysis, or other means).

[0024] The application of the novel compound containing a biphenylamide analog structure provided by the present invention, its pharmaceutically acceptable salt, or pharmaceutical composition in the preparation of drugs for treating diseases mediated by EZH2, PRC2, or EZH2 / PRC2.

[0025] Furthermore, the application of the novel compound containing a biphenylamide analog structure provided by the present invention, its pharmaceutically acceptable salt, or pharmaceutical composition in the preparation of drugs with EZH2 as a molecular therapeutic target.

[0026] Furthermore, the application of the novel compound containing a biphenylamide analog structure provided by the present invention, its pharmaceutically acceptable salt, or pharmaceutical composition in the preparation of anti-tumor drugs. The types of tumors mainly include liver cancer, nasopharyngeal carcinoma, colorectal cancer, melanoma, bladder cancer, leukemia, esophageal cancer, breast cancer, gastric cancer, prostate cancer, pancreatic cancer, lung cancer, ovarian cancer, epithelioid sarcoma, non-Hodgkin lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, and follicular lymphoma, etc.

[0027] The beneficial effects of the present invention are:

[0028] 1. The novel compounds containing a biphenylamide analogue structure provided by the present invention and their pharmaceutically acceptable salts have excellent anti-tumor activity and safety. They can be used for the treatment of neoplastic diseases such as epithelioid sarcoma, lymphoma, prostate cancer, and breast cancer, and other autoimmune diseases.

[0029] 2. The novel compounds containing a biphenylamide analogue structure provided by the present invention and their pharmaceutically acceptable salts, through the EZH2 enzyme activity test, are found to have significant inhibitory activity on EZH2 kinase activity, and are particularly used for the use in the preparation of drugs for the treatment and / or prevention of diseases caused by abnormal overexpression of EZH2 kinase, especially for the use in the preparation of drugs for the treatment and / or prevention of cancer.

[0030] 3. The novel compounds containing a biphenylamide analogue structure provided by the present invention have novel chemical structures. Most of the compounds have significant inhibitory activity on EZH2 kinase in in vitro biological activity studies and strong anti-proliferative activity on the tested cells. The compounds can be used for the treatment and prevention of cancer. Detailed Embodiments

[0031] The examples and preparation examples provided below further illustrate and exemplify the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparation examples does not limit the scope of the present invention in any way.

[0032] The following synthetic routes describe the preparation methods of the novel compounds containing a biphenylamide analogue structure of general formula (I) or (II) of the present invention.

[0033] All raw materials are prepared by the methods known to those of ordinary skill in the art of organic chemistry or are commercially available in the manner described by the following synthetic routes. All final compounds of the present invention are prepared by the methods described by the following synthetic routes or by methods similar thereto, which are known to those of ordinary skill in the art of organic chemistry. All variable factors applied in the following synthetic routes are defined as below or as defined in the claims.

[0034] The examples are intended to illustrate rather than limit the scope of the present invention. The nuclear magnetic resonance hydrogen spectrum of the compounds was measured with a Bruker ARX-400 or ARX-600, and the mass spectrum was measured with an Agilent 1100 LC / MSD; all reagents used were of analytical grade or chemical pure grade.

[0035] According to the present invention, the synthetic routes of Compounds 1 to 34 of general formula (I) or (II) are as follows, and the functional groups of A, B, X, and R are as shown in the groups in the claims.

[0036] Example 1: N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(4-methoxypiperidin-1-yl)quinolin-7-yl)-1H-indazole-4-carboxamide (Compound 1)

[0037]

[0038] Step 1: Synthesis of methyl 6-bromo-1-isopropyl-1H-indazole-4-carboxylate

[0039]

[0040] Dissolve methyl 6-bromo-1H-indazole-4-carboxylate (15 g, 58.8 mmol) in 60 mL of ACN. Add 1.5 eq of iodopropane (15 g, 88.2 mmol) and 2 eq of Cs2CO3 (38.4 g), and mix well. Place the mixture in an oil bath at 90 °C, connect a reflux tube, and react for 4 h. Monitor the reaction by TLC until completion. Filter off the solid in the reaction solution, wash it with a small amount of DCM to obtain a filtrate. Add 100 mL of DCM to the filtrate, wash it three times with 300 mL of water in portions, and then wash it twice with saturated NaCl solution, 30 mL each time. Then, completely dry the organic phase with anhydrous sodium sulfate and perform vacuum distillation to obtain an orange-yellow oil, which is then cooled and crystallized to obtain 17 g of solid. The yield is 97.6%.

[0041] Step 2: Synthesis of methyl 1-isopropyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole-4-carboxylate

[0042]

[0043] Add potassium acetate 2eq (11.3 g) and 150 mL of Diox into a three-necked flask, and bubble N2 for 15 min; add methyl 6-bromo-1-isopropyl-1H-indazole-4-carboxylate (17 g, 57.4 mmol) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) 2eq (29.2 g, 11.5 mmol), and bubble N2 for 15 min; add Pd(dppf)Cl2 0.05eq (2.1 g), evacuate three times, place it in an oil bath at 80 °C under nitrogen protection, connect the reflux tube and react for 6 h, monitor by TLC, and the reaction is complete. Filter out the solid in the reaction solution, wash it with a small amount of DCM to obtain the filtrate; add 100 mL of DCM and 300 mL of water to the filtrate and wash it 3 times, then wash it twice with saturated NaCl solution, 30 mL each time. Then use anhydrous sodium sulfate to completely dry the organic phase, distill it under reduced pressure, and purify it by column chromatography to obtain 11.423 g of a pale yellow solid, with a yield of 57.8%.

[0044] Step 3: Synthesis of 7-bromo-4-(4-methoxypiperidin-1-yl)quinoline

[0045]

[0046] Mix 7-bromo-4-chloroquinoline (1 g, 4.12 mmol) with 10 mL of anhydrous DMF solution, then add K2CO3 3eq (1.7 g) and mix well, add 4-methoxypiperidine 1.5eq (0.72 g, 6.25 mmol), and then react at 80 °C in an oil bath for 24 h, monitor by TLC, and the reaction is complete. Filter out the solid in the reaction solution, wash it with a small amount of DCM to obtain the filtrate; add 100 mL of DCM and 300 mL of water to the filtrate and wash it 3 times, then wash it twice with saturated NaCl solution, 30 mL each time. Then use anhydrous sodium sulfate to completely dry the organic phase, distill it under reduced pressure, and purify it by column chromatography to obtain 800 mg, with a yield of 61.5%.

[0047] Step 4: Synthesis of methyl 1-isopropyl-6-[4-(4-methoxypiperidin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxylate

[0048]

[0049] Add potassium acetate 2eq (1.84 g) and 30 mL of Diox into a three-necked flask, and blow N2 for 15 min; add 1.5eq (4.8 g, 13.9 mmol) of methyl 1-isopropyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole-4-carboxylate and 7-bromo-4-(4-methoxypiperidin-1-yl)quinoline (3 g, 9.3 mmol), and blow N2 for 15 min; add 0.05eq (0.55 g) of Pd(dppf)Cl2, evacuate three times, place it in an oil bath at 80 °C under nitrogen protection, connect the reflux tube and react for 6 h. Monitor by TLC, and the reaction is complete. Filter out the solid in the reaction solution, wash it with a small amount of DCM to obtain the filtrate; add 100 mL of DCM and 300 mL of water to wash it three times, and then wash it twice with saturated NaCl solution, 30 mL each time. Then use anhydrous sodium sulfate to completely dry the organic phase, distill it under reduced pressure, and then purify it by column chromatography to obtain 564 mg of solid, with a yield of 13.1%.

[0050] Step 5: Synthesis of 1-isopropyl-6-[4-(4-methoxypiperidin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxylic acid

[0051]

[0052] Mix methyl 1-isopropyl-6-[4-(4-methoxypiperidin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxylate with 5V / mol of EtOH and 5V / mol of water, and after mixing evenly, add 1.5eq of NaOH to the mixture. Place the reaction solution in an oil bath at 60 °C, connect the reflux and react for 6 h. Monitor by TLC, and the reaction is complete. Then rotary evaporate part of the ethanol solution in the reaction solution, add 10 mL of H2O, extract it once with DCM, take the aqueous phase, adjust the pH to acidic with dilute hydrochloric acid diluted 10 times, precipitate the solid, and filter it to obtain 378 mg, with a yield of 75%.

[0053] Step 6: Synthesis of N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-1-isopropyl-6-[4-(4-methoxypiperidin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxamide (Compound 1)

[0054]

[0055] 1-Isopropyl-6-[4-(4-methoxypiperidin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxylic acid (100 mg, 22.5 mmol) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one 1.1 eq (37.7 mg) were mixed and dissolved in 5 mL of DMF. Then, HATU 2 eq (171.2 mg) and TEA 3 eq (68.24 mg) were added. The reaction was carried out at room temperature for 4 h and monitored by TLC until the reaction was complete. The mixture was extracted three times with 20 mL of DCM to obtain the organic phase. The organic phase was washed three times with 50 mL of water and then washed twice with saturated NaCl solution, 20 mL each time, to further wash the organic phase. Then, the organic phase was completely dried with anhydrous sodium sulfate. Sodium sulfate was removed by suction filtration. The filtrate was distilled under reduced pressure until completely dry. Then, it was purified by PTLC to obtain 35 mg of Compound 1, with a yield of 30.7%.

[0056] MS(ESI), m / z: 579.30 [M+H] + , 601.30 [M+Na] + ; 1 1H NMR (500 MHz, CDCl3) δ 8.69 (d, J =

[0057] 5.0 Hz, 1H), 8.26–8.20 (m, 1H), 8.04 (d, J = 8.7 Hz, 1H), 7.74 (d, J = 2.1 Hz, 1H), 7.69 (dd, J = 8.7, 1.9 Hz, 1H), 7.58 (d, J = 2.0 Hz, 1H), 7.43 (s, 1H), 6.82 (d, J = 5.0 Hz, 1H), 5.85 (s, 1H), 4.55 (d, J = 5.9 Hz, 2H), 4.15–3.98 (m, 3H), 3.43 (d, J = 11.9 Hz, 2H), 3.31 (s, 1H), 2.63 (t, J = 11.1 Hz, 2H), 2.35 (d, J = 4.6 Hz, 6H), 2.12 (s, 3H), 1.27 (d, J = 6.3 Hz, 6H), 1.16–1.10 (m, 4H), 1.04–0.93 (m, 2H), 0.63–0.52 (m, 2H).

[0058] By replacing appropriate raw materials and reagents and following the preparation scheme of Example 1, Examples 2-10, Example 12, Examples 21-22, and Examples 30-34 (Compounds 2-10, Compound 12, Compounds 21-22, and Compounds 30-34) were finally prepared.

[0059] Example 2: N-(2,6-dioxopiperidin-3-yl)-1-isopropyl-6-[4-(4-methoxypiperidin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxamide (Compound 2)

[0060]

[0061] MS(ESI), m / z: 555.20 [M+H] + , 577.20 [M+Na] + ; 1 H NMR(500 MHz, CDCl3) δ 8.67 (s, 1H), 8.51 (s, 1H), 8.38 (s, 2H), 8.13–7.86 (m, 4H), 6.80 (s, 1H), 4.96 (d, J = 6.1 Hz, 2H), 3.49 (d, J = 54.0 Hz, 6H), 3.14 (s, 2H), 2.94–2.70 (m, 3H), 2.15 (s, 3H), 1.93 (s, 2H), 1.69–1.59 (m, 6H), 1.42 (s, 1H), 1.25 (s, 1H).

[0062] Example 3: N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-1-isopropyl-6-[4-(4-methylpiperidin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxamide (Compound 3)

[0063]

[0064] MS(ESI), m / z: 563.32 [M+H] + ; 1 H NMR(500 MHz, CDCl3) δ 12.40 (s, 1H), 8.65 (d, J = 5.2

[0065] Hz, 1H), 8.46 (s, 1H), 8.34 (s, 1H), 8.09–8.02 (m, 2H), 7.92 (s, 1H), 7.87 (s, 1H), 7.81 (dd, J=8.7, 2.0 Hz, 1H), 6.81 (d, J=5.3 Hz, 1H), 5.84 (s, 1H), 4.93 (p, J=6.6 Hz, 1H), 4.65 (d, J=5.8 Hz, 2H), 3.67 (d, J=12.4 Hz, 2H), 2.90 (t, J=11.3 Hz, 2H), 2.38 (s, 3H), 2.12 (s, 3H), 1.85 (dd, J=13.4, 2.6 Hz, 2H), 1.61 (d, J=6.7 Hz, 6H), 1.55 (dd, J=12.1, 3.5 Hz, 2H), 1.25 (s, 1H), 1.07 (d, J=6.4 Hz, 3H).

[0066] Example 4: N-(2,6-Dioxopiperidin-3-yl)-1-isopropyl-6-[4-(4-methylpiperidin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxamide (Compound 4)

[0067]

[0068] MS(ESI), m / z: 539.20 [M+H] + ; 1 1H NMR (500 MHz, CDCl3) δ 12.36 (s, 1H), 8.70 (d, J=5.0

[0069] Hz, 1H), 8.45 (s, 1H), 8.34 (s, 1H), 8.04 (d, J=8.7 Hz, 1H), 7.92 (s, 1H), 7.87–7.82 (m, 2H), 6.82 (d, J=5.0 Hz, 1H), 5.86 (s, 1H), 4.93 (p, J=6.6 Hz, 1H), 4.65 (d, J=5.8 Hz, 2H), 4.03 (t, J=8.3 Hz, 2H), 3.47 (d, J=11.9 Hz, 2H), 2.66 (t, J=11.2 Hz, 2H), 2.39 (s, 3H), 2.12 (s, 3H), 1.61 (d, J=6.7 Hz, 6H), 1.28 (s, 3H).

[0070] Example 5: N-[2-(1H-Indol-3-yl)ethyl]-1-isopropyl-6-[4-(4-methylpiperidin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxamide (Compound 5)

[0071]

[0072] MS(ESI), m / z: 571.20 [M+H] + , 593.20 [M+Na] + ; 1 H NMR(500 MHz, CDCl3) δ 9.57(s, 1H), 8.72(d, J=5.3 Hz, 1H), 8.39–8.31(m, 2H), 8.06(d, J=8.7 Hz, 1H), 7.85–7.76(m, 2H), 7.69–7.64(m, 2H), 7.41(d, J=8.1 Hz, 1H), 7.29(d, J=2.3 Hz, 1H), 7.14(dt, J=31.5, 7.3 Hz, 2H), 6.85(d, J=5.3 Hz, 1H), 6.42(d, J=5.9 Hz, 1H), 4.92(p, J=6.6 Hz, 1H), 3.90(q, J=6.2 Hz, 2H), 3.71(d, J=12.2 Hz, 2H), 3.20(t, J=6.4 Hz, 2H), 2.98–2.90(m, 2H), 1.91–1.85(m, 2H), 1.62(d, J=6.7 Hz, 9H), 1.09(d, J=6.4 Hz, 3H).

[0073] Example 6: N-[2-(1H-imidazol-4-yl)ethyl]-1-isopropyl-6-[4-(4-methylpiperidin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxamide (Compound 6)

[0074]

[0075] MS(ESI), m / z: 522.20 [M+H] + ; 1 H NMR(400 MHz, CDCl3) δ 8.59(d, J=5.2 Hz, 1H), 8.42

[0076] (s, 1H), 8.27 (s, 1H), 8.09 (d, J = 2.1 Hz, 1H), 7.99 (d, J = 8.7 Hz, 1H), 7.86 (s, 1H), 7.80 (d, J = 8.7 Hz, 3H), 7.71 (s, 1H), 6.92 (s, 1H), 6.76 (d, J = 5.3 Hz, 1H), 4.90 (p, J = 6.7 Hz, 1H), 3.89–3.76 (m, 2H), 3.62 (d, J = 11.9 Hz, 2H), 2.98 (t, J = 6.3 Hz, 2H), 2.85 (t, J = 11.7 Hz, 2H), 1.83 (d, J = 12.4 Hz, 2H), 1.60 (d, J = 6.7 Hz, 6H), 1.35–1.22 (m, 4H), 1.06 (d, J = 6.2 Hz, 3H).

[0077] Example 7: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-1-isopropyl-6-[4-(4-methylpiperazin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxamide (Compound 7)

[0078]

[0079] MS(ESI), m / z: 564.20 [M+H] + ; 1 1H NMR (500 MHz, CDCl3) δ 12.39 (s, 1H), 8.70 (d, J = 5.0

[0080] Hz, 1H), 8.46 (s, 1H), 8.34 (d, J = 2.0 Hz, 1H), 8.06 (dd, J = 12.0, 7.2 Hz, 2H), 7.95–7.91 (m, 1H), 7.86 (s, 1H), 7.82 (dd, J = 8.7, 1.9 Hz, 1H), 6.85 (d, J = 5.1 Hz, 1H), 5.84 (s, 1H), 4.92 (p, J = 6.7 Hz, 1H), 4.65 (d, J = 5.7 Hz, 2H), 3.34 (t, J = 4.8 Hz, 4H), 2.79 (t, J = 4.7 Hz, 4H), 2.47 (s, 3H), 2.38 (s, 3H), 2.12 (s, 3H), 1.61 (d, J = 6.7 Hz, 6H).

[0081] Example 8: N-(2,6-Dioxopiperidin-3-yl)-1-isopropyl-6-[4-(4-methylpiperazin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxamide (Compound 8)

[0082]

[0083] MS(ESI), m / z: 540.30 [M+H] + ; 1 H NMR(400 MHz, CDCl3) δ 8.66(s, 2H), 8.51(s, 1H), 8.38(s, 1H), 8.11–8.05(m, 2H), 8.01(s, 1H), 7.89(s, 2H), 6.80(s, 1H), 4.98–4.91(m, 1H), 3.76–3.70(m, 1H), 3.40(s, 4H), 2.77(s, 4H), 2.46(s, 3H), 1.63(d, J = 7.7 Hz, 4H), 1.26–1.23(m, 6H).

[0084] Example 9: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-6-[4-((2S,6R)-2,6-dimethylmorpholino)quinolin-7-yl]-1-isopropyl-1H-indazole-4-carboxamide (Compound 9)

[0085]

[0086] MS(ESI), m / z: 579.20 [M+H] + ; 1 H NMR(500 MHz, CDCl3) δ 11.81(s, 1H), 8.69(d, J = 5.0

[0087] Hz, 1H), 8.26–8.20(m, 1H), 8.04(d, J = 8.7 Hz, 1H), 7.79–7.66(m, 2H), 7.60–7.39(m, 2H), 6.82(d, J = 5.0 Hz, 1H), 5.85(s, 1H), 4.55(s, 2H), 4.03(ddd, J = 10.4, 6.2, 2.1 Hz, 2H), 3.43(d, J = 11.9 Hz, 2H), 3.31(dq, J = 14.1, 7.1 Hz, 1H), 2.63(t, J = 11.1 Hz, 2H), 2.35(d, J = 4.6 Hz, 6H), 2.12(s, 3H), 1.27(d, J = 6.3 Hz, 6H), 1.16–1.11(m, 3H).

[0088] Example 10: 6-[4-((2S,6R)-2,6-dimethylmorpholino)quinolin-7-yl]-N-(2,6-dioxopiperidin-3-yl)-1-isopropyl-1H-indazole-4-carboxamide (Compound 10)

[0089]

[0090] MS(ESI), m / z: 555.20 [M+H] + ; 1 H NMR(500 MHz, CDCl3) δ 8.78 (s, 1H), 8.51 (s, 1H), 8.42 (s, 1H), 8.18 (s, 1H), 8.12 (d, J = 8.6 Hz, 1H), 7.97 (d, J = 15.8 Hz, 2H), 7.87 (d, J = 8.6 Hz, 1H), 6.86 (s, 1H), 5.00–4.87 (m, 2H), 4.06 (s, 2H), 3.49 (d, J = 11.9 Hz, 2H), 2.86 (d, J = 23.6 Hz, 2H), 2.68 (t, J = 11.0 Hz, 2H), 1.66 (d, J = 7.4 Hz, 6H), 1.30 (d, J = 6.2 Hz, 6H), 1.13 (d, J = 6.1 Hz, 2H).

[0091] Example 11: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-[4-((2S,6R)-2,6-dimethylmorpholinyl)quinolin-8-yl]-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzamide (Compound 11)

[0092] Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic acid

[0093]

[0094] Using 2-methyl-3-nitrobenzoic acid (59 g, 325.9 mmol) as the raw material, it was fully mixed with 200 mL of concentrated H2SO4 under low temperature conditions, and 0.55 eq (51 g, 178.3 mmol) of 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione was added under stirring. The reaction was carried out at room temperature for 6 h, monitored by TLC, and the reaction was complete. The reaction solution was added to cold water, and a white powdery solid was precipitated. It was filtered by suction to obtain 84 g, with a yield of 99%.

[0095] Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate

[0096]

[0097] 5-Bromo-2-methyl-3-nitrobenzoic acid (15 g, 57.9 mmol) was added to 80 mL of 5 V / mol CH3OH. After thorough mixing, 25 mL of concentrated H2SO4 was added dropwise to the mixture. The reaction was carried out in an oil bath at 60 °C with a reflux condenser for 4 h. The reaction was monitored by TLC and was complete. The reaction mixture was rotary evaporated to remove excess CH3OH, and then poured into 20% Na2CO3 solution. The pH was adjusted to 9 with NaOH, and the mixture was allowed to stand to obtain a precipitate, which was filtered by suction to give 13.3 g of a white solid with a yield of 84%.

[0098] Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate

[0099]

[0100] In a mixed solution of EA:H2O = 5:1, while stirring, methyl 5-bromo-2-methyl-3-nitrobenzoate (13 g, 47.6 mmol), 5 eq of Fe powder (13.3 g, 273.5 mmol), and 15 eq of CH3COOH (43 g) were added successively. The reaction was carried out in an oil bath at 80 °C with a reflux condenser for 6 h. The reaction was monitored by TLC and was complete. The reaction mixture was allowed to cool to room temperature and filtered by suction to obtain a filtrate. The pH of the filtrate was adjusted to 9 with NaOH, and then extracted with EA. The organic phase was washed 3 times with 150 mL of water and then washed 2 times with 50 mL of saturated Na2CO3 solution to further purify the reaction mixture. Then, the organic phase was completely dried with anhydrous sodium sulfate. The sodium sulfate was removed by suction filtration. The filtrate was distilled under reduced pressure until completely dry. 9 g of a grayish-red solid was obtained with a yield of 78%.

[0101] Step 4: Synthesis of methyl 5-bromo-3-(cyclopropanecarboxamido)-2-methylbenzoate

[0102]

[0103] Methyl 3-amino-5-bromo-2-methylbenzoate (5 g, 20.6 mmol), DMAP 0.2 eq (0.5 g), and DIPEA 3.5 eq (9.3 g) were added to a sealed reaction apparatus. Then, 20 mL of THF solution was injected with a syringe and mixed evenly. Subsequently, 1.4 eq (3 g, 28.8 mmol) of cyclopropanecarbonyl chloride dissolved and diluted with THF was added dropwise. Be careful not to expose to air for a long time. Then, the reaction was carried out at room temperature for 4 h. Monitored by TLC, the reaction was complete. The reaction solution was left at room temperature. The solid in the reaction solution was removed by suction filtration, and the filter cake was washed with a small amount of DCM. The filtrate was obtained. 60 mL of DCM was added to the filtrate in three portions for extraction to obtain the organic phase. The organic phase was washed three times with 100 mL of water, and then washed twice with saturated NaCl solution, 30 mL each time, to further purify the organic phase. Then, anhydrous sodium sulfate was used to completely dry the organic phase. Sodium sulfate was removed by suction filtration. The filtrate was distilled under reduced pressure until completely dry. 6 g of brown solid was obtained, with a yield of 93.7%.

[0104] Step 5: Synthesis of methyl 5-bromo-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzoate

[0105]

[0106] Methyl 5-bromo-3-(cyclopropanecarboxamido)-2-methylbenzoate (12.86 g, 41.3 mmol) was mixed with 60 mL of ACN until dissolved. Iodoethane 1.2 eq (7.73 g, 49.5 mmol) and Cs2CO3 2 eq (27 g) were added and mixed evenly. The mixture was placed in an oil bath at 90 °C and connected to a reflux tube for reaction for 4 h. Monitored by TLC, the reaction was complete. The reaction system was cooled to room temperature. The solid in the reaction solution was removed by suction filtration, and the filter cake was washed with a small amount of DCM. The filtrate was obtained. 100 mL of DCM was added to the filtrate in three portions for extraction to obtain the organic phase. The organic phase was washed three times with 300 mL of water, and then washed twice with saturated NaCl solution, 30 mL each time, to further purify the organic phase. Then, anhydrous sodium sulfate was used to completely dry the organic phase. Sodium sulfate was removed by suction filtration. The filtrate was distilled under reduced pressure until completely dry to obtain an orange-yellow oil, which was then cooled and crystallized to obtain 13.375 g of pale yellow solid, with a yield of 95.5%.

[0107] Step 6: Synthesis of methyl 3-(N-ethylcyclopropanecarboxamido)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate

[0108]

[0109] Add potassium acetate 2eq (6.32 g) and 100 mL of Diox into a three-necked flask, and bubble N2 for 15 min; add methyl 5-bromo-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzoate (9.2 g, 27 mmol) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) 2eq (13.8 g, 54.3 mmol), and bubble N2 for 15 min; add Pd(PPh3)Cl2 0.05eq (0.96 g), evacuate the vacuum three times, and place it in an oil bath at 100 °C under nitrogen protection, connect the reflux tube and react for 6 h. Monitor by TLC, and the reaction is complete. Cool the reaction system to room temperature, remove the solid in the reaction solution by suction filtration, wash the filter cake with a small amount of DCM, and obtain the filtrate; add 100 mL of DCM to the filtrate in 3 portions for extraction to obtain the organic phase. Wash the organic phase with 300 mL of water in 3 portions, and then wash it twice with saturated NaCl solution, 30 mL each time, to further purify the organic phase. Then, use anhydrous sodium sulfate to completely dry the organic phase. Remove sodium sulfate by suction filtration. Distill the filtrate under reduced pressure until completely dry. Then, purify it by column chromatography to obtain 8.84 g of a pale yellow solid, with a yield of 84.2%.

[0110] Step 7: Synthesis of (2S,6R)-4-(7-bromoquinolin-4-yl)-2,6-dimethylmorpholine

[0111]

[0112] Mix 7-bromo-4-chloro-quinoline (1 g, 4.12 mmol) with 10 mL of anhydrous DMF solution, then add K2CO3 3eq (1.7 g) and mix well. Add (2S,6R)-2,6-dimethyl-4-morpholine 1.5eq (0.72 g, 6.25 mmol), and then react in an oil bath at 80 °C for 24 h. Monitor by TLC, and the reaction is complete. Cool the reaction system to room temperature, remove the solid in the reaction solution by suction filtration, wash the filter cake with a small amount of DCM, and obtain the filtrate; add 20 mL of DCM to the filtrate in 3 portions for extraction to obtain the organic phase. Wash the organic phase with 50 mL of water in 3 portions, and then wash it twice with saturated NaCl solution, 20 mL each time, to further purify the organic phase. Then, use anhydrous sodium sulfate to completely dry the organic phase. Remove sodium sulfate by suction filtration. Distill the filtrate under reduced pressure until completely dry, and then purify it by column chromatography to obtain 970 mg of a pale yellow solid, with a yield of 74.6%.

[0113] Step 8: Synthesis of methyl 5-[4-((2S,6R)-2,6-dimethylmorpholinyl)quinolin-7-yl]-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzoate

[0114]

[0115] Add potassium acetate 2eq (0.56 g) and 10 mL of Diox into a three-necked flask, and bubble N2 for 15 min; add methyl 3-(N-ethylcyclopropanecarboxamido)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (1 g, 2.6 mmol) and (2S,6R)-4-(7-bromoquinolin-4-yl)-2,6-dimethylmorpholine 1.1eq (1 g), and bubble N2 for 15 min; add Pd(PPh3)Cl2 0.05eq (0.01 g), evacuate three times, place it in an oil bath at 100 °C under nitrogen protection, connect the reflux tube and react for 6 h, monitor by TLC, and the reaction is complete. Cool the reaction system to room temperature, remove the solid in the reaction solution by suction filtration, wash the filter cake with a small amount of DCM, and obtain the filtrate; add 100 mL of DCM to the filtrate in three portions for extraction to obtain the organic phase, wash the organic phase with 300 mL of water in three portions, and then wash it twice with saturated NaCl solution, 30 mL each time, to further purify the organic phase. Then use anhydrous sodium sulfate to completely dry the organic phase. Remove sodium sulfate by suction filtration. Distill the filtrate under reduced pressure until completely dry. Column chromatography gives 1 g, with a yield of 69.7%.

[0116] Step 9: Synthesis of 5-(4-((2S,6R)-2,6-dimethylmorpholino)quinolin-7-yl)-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzoic acid

[0117]

[0118] Mix the above-obtained methyl 5-[4-((2S,6R)-2,6-dimethylmorpholino)quinolin-7-yl]-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzoate with 5 V / mol of EtOH and 5 V / mol of water, and after mixing evenly, add 1.5eq of NaOH to the mixture, place the reaction solution in an oil bath at 60 °C, connect the reflux and react for 6 h, monitor by TLC, and the reaction is complete. Then rotary evaporate part of the ethanol solution in the reaction solution, add 10 mL of H2O, extract once with DCM, take the aqueous phase, adjust the pH to acidic with dilute hydrochloric acid diluted 10 times, precipitate the solid, and filter by suction to obtain 806 mg, with a yield of 87%.

[0119] Step 10: Synthesis of N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-[4-((2S,6R)-2,6-dimethylmorpholino)quinolin-8-yl]-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzamide (Compound 11)

[0120]

[0121] Using 5-(4-((2S,6R)-2,6-dimethylmorpholino)quinolin-7-yl)-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzoic acid and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one as raw materials, they were mixed and dissolved in 5 mL of DMF. Then, 2 eq (171.2 mg) of HATU and 3 eq (68.24 mg) of TEA were added. The reaction was carried out at room temperature for 4 h and monitored by TLC until the reaction was complete. The mixture was extracted three times with 20 mL of DCM to obtain the organic phase. The organic phase was washed three times with 50 mL of water and then washed twice with saturated NaCl solution, 20 mL each time, to further wash the organic phase. Then, anhydrous sodium sulfate was used to completely dry the organic phase. Sodium sulfate was removed by suction filtration. The filtrate was distilled under reduced pressure until completely dry. Then, it was purified by PTLC to obtain 75 mg of compound 11 with a yield of 39.4%.

[0122] MS(ESI), m / z: 622.20[M+H] + ; 1 H NMR(500 MHz, CDCl3)δ11.86(s, 1H), 8.73(d, J = 5.0

[0123] Hz, 1H), 8.26–8.22(m, 1H), 8.06(d, J = 8.7 Hz, 1H), 7.76–7.67(m, 2H), 7.58(d, J = 2.0 Hz, 1H), 7.39(t, J = 5.9 Hz, 1H), 6.85(d, J = 5.1 Hz, 1H), 5.85(s, 1H), 4.55(d, J = 5.8 Hz, 2H), 3.99(t, J = 4.5 Hz, 4H), 3.26(t, J = 4.5 Hz, 4H), 2.35(d, J = 6.1 Hz, 7H), 2.13(s, 3H), 1.34–1.08(m, 9H), 1.03–0.94(m, 2H), 0.65–0.50(m, 2H).

[0124] By replacing the appropriate raw materials and reagents and following the preparation scheme of Example 11, Examples 13 - 20 and Examples 23 - 29 (compounds 13 - 20, compounds 23 - 29) were finally prepared.

[0125] Example 12: 1-Cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((2S,6R)-2,6-dimethylmorpholino)quinolin-7-yl)-1H-indazole-4-carboxamide (Compound 12)

[0126]

[0127] MS(ESI), m / z: 605.30 [M+H] + ; 1 H NMR(500 MHz, CDCl3) δ 12.10 (s, 1H), 8.66 (s, 1H), 8.58 (s, 1H), 8.36 (s, 1H), 8.17 (d, J = 15.4 Hz, 2H), 8.03 (d, J = 8.6 Hz, 1H), 7.92–7.79 (m, 2H), 6.79 (d, J = 5.0 Hz, 1H), 5.83 (d, J = 14.2 Hz, 1H), 4.94 (dp, J = 14.6, 7.3 Hz, 1H), 4.62 (d, J = 5.4 Hz, 2H), 4.03 (d, J = 9.0 Hz, 2H), 3.48 (d, J = 11.9 Hz, 2H), 2.66 (t, J = 11.2 Hz, 2H), 2.36 (d, J = 7.3 Hz, 3H), 2.32–2.27 (m, 2H), 2.20 (dd, J = 13.5, 7.0 Hz, 2H), 2.15 (s, 3H), 1.95 (d, J = 7.7 Hz, 3H), 1.75 (t, J = 6.0 Hz, 3H), 1.27 (d, J = 6.2 Hz, 6H).

[0128] Example 13: N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(N-ethylcyclopropanecarboxamido)-2-methyl-5-(4-morpholinylquinolin-8-yl)benzamide (Compound 13)

[0129]

[0130] MS(ESI), m / z: 594.20 [M+H] + , 616.20 [M+Na] + ; 11H NMR (400 MHz, DMSO) δ 10.98 (s, 1H), 9.13 (d, J = 8.4 Hz, 1H), 8.71 (d, J = 5.0 Hz, 1H), 8.52 (d, J = 1.6 Hz, 1H), 8.44 (d, J = 8.1 Hz, 2H), 8.17 (d, J = 1.2 Hz, 1H), 8.13–8.06 (m, 2H), 6.96 (d, J = 5.0 Hz, 1H), 5.28 (p, J = 6.5 Hz, 1H), 4.93 (ddd, J = 13.0, 8.3, 5.4 Hz, 1H), 3.56 (d, J = 11.7 Hz, 2H), 3.36 (s, 1H), 2.92–2.77 (m, 3H), 2.63–2.48 (m, 4H), 2.26–2.03 (m, 2H), 1.81 (dd, J = 12.7, 3.4 Hz, 2H), 1.66–1.43 (m, 10H), 1.02 (d, J = 6.2 Hz, 3H).

[0131] Example 14: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-(N-ethylcyclopropanecarboxamido)-2-methyl-5-[4-(4-methylpiperazin-1-yl)quinolin-8-yl]benzamide (Compound 14)

[0132]

[0133] MS (ESI), m / z: 607.30 [M+H] + ,629.20 [M+Na] + ; 1 1H NMR (500 MHz, CDCl3) δ 11.91 (s, 1H), 8.66 (s, 1H), 8.22 (s, 1H), 8.03 (s, 1H), 7.78–7.64 (m, 2H), 7.59–7.47 (m, 2H), 6.83 (d, J = 5.2 Hz, 1H), 5.85 (s, 1H), 4.54 (d, J = 6.0 Hz, 2H), 4.15–4.05 (m, 1H), 3.39–3.25 (m, 5H), 2.73 (s, 4H), 2.43 (s, 4H), 2.35 (d, J = 3.8 Hz, 6H), 2.13 (s, 3H), 1.13 (t, J = 7.1 Hz, 3H), 0.98 (d, J = 17.9 Hz, 2H), 0.65–0.50 (m, 2H).

[0134] Example 15: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-(N-ethylcyclopropanecarboxamido)-5-[4-(4-ethylpiperazin-1-yl)quinolin-8-yl]-2-methylbenzamide (Compound 15)

[0135]

[0136] MS(ESI), m / z: 621.30 [M+H] + , 643.30 [M+Na] + ; 1 1H NMR (500 MHz, CDCl3) δ 11.94 (s, 1H), 8.23 (d, J = 5.0 Hz, 1H), 8.05 (d, J = 8.6 Hz, 1H), 7.74 (s, 1H), 7.67 (s, 1H), 7.57 (s, 1H), 7.47 (d, J = 10.9 Hz, 1H), 6.84 (d, J = 4.8 Hz, 1H), 5.91–5.81 (m, 1H), 4.55 (d, J = 5.5 Hz, 2H), 4.10 (qd, J = 7.4, 3.6 Hz, 1H), 3.33 (dt, J = 14.0, 6.0 Hz, 5H), 2.79 (s, 4H), 2.65–2.55 (m, 2H), 2.36 (q, J = 3.6 Hz, 6H), 2.14 (td, J = 8.6, 3.6 Hz, 3H), 1.22–1.11 (m, 6H), 0.98 (d, J = 22.0 Hz, 2H), 0.58 (d, J = 23.6 Hz, 2H).

[0137] Example 16: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-(N-ethylcyclopropanecarboxamido)-5-[4-(3-isopropoxyazetidin-1-yl)quinolin-8-yl]-2-methylbenzamide (Compound 16)

[0138]

[0139] MS(ESI), m / z: 622.30 [M+H] + ; 1 1H NMR (500 MHz, CDCl3) δ 8.41 (d, J = 16.0 Hz, 1H),

[0140] 8.31 (s, 1H), 8.25 (s, 1H), 7.99 (t, J = 8.1 Hz, 1H), 7.81 (s, 1H), 7.70–7.49 (m, 3H), 6.21 (d, J = 16.9 Hz, 1H), 5.90 (s, 1H), 4.75 (d, J = 36.3 Hz, 2H), 4.62 (d, J = 10.3 Hz, 1H), 4.52 (s, 1H), 4.36 (d, J = 28.6 Hz, 2H), 4.06 (dd, J = 13.7, 7.0 Hz, 2H), 3.80–3.63 (m, 2H), 3.37 (dd, J = 13.3, 7.0 Hz, 1H), 3.04 (s, 1H), 2.38 (d, J = 13.2 Hz, 5H), 2.25 (d, J = 7.5 Hz, 3H), 1.22 (d, J = 6.2 Hz, 8H), 1.17–1.11 (m, 3H), 0.59 (d, J = 31.6 Hz, 2H).

[0141] Example 17: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-(N-ethylcyclopropanecarboxamido)-2-methyl-5-[4-(4-methylsulfonylpiperazin-1-yl)quinolin-8-yl]benzamide (Compound 17)

[0142]

[0143] MS (ESI), m / z: 671.30 [M + H] + ,693.20 [M + Na] + ; 1 1H NMR (500 MHz, CDCl3) δ 11.60 (s, 1H), 8.75 (d, J = 5.0 Hz, 1H), 8.26 (d, J = 1.9 Hz, 1H), 8.02 (d, J = 8.7 Hz, 1H), 7.76–7.69 (m, 2H), 7.57 (d, J = 2.0 Hz, 1H), 7.42 (t, J = 5.9 Hz, 1H), 6.89 (d, J = 5.0 Hz, 1H), 5.88 (s, 1H), 4.55 (d, J = 5.9 Hz, 2H), 4.15–4.05 (m, 1H), 3.55 (t, J = 4.8 Hz, 4H), 3.41–3.29 (m, 5H), 2.91 (s, 3H), 2.36 (d, J = 13.3 Hz, 7H), 2.17 (s, 3H), 1.14 (t, J = 7.1 Hz, 3H), 1.05–0.92 (m, 2H), 0.65–0.50 (m, 2H).

[0144] Example 18: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-(N-ethylcyclopropanecarboxamido)-5-[4-(4-(2-methoxyethyl)piperazin-1-yl)quinolin-8-yl]-2-methylbenzamide (Compound 18)

[0145]

[0146] MS(ESI), m / z: 651.30 [M+H] + , 673.30 [M+Na] + ; 1 1H NMR (400 MHz, CDCl3) δ 11.93 (s, 1H), 8.69 (d, J = 5.0 Hz, 1H), 8.21 (s, 1H), 8.05 (d, J = 8.8 Hz, 1H), 7.72 (s, 1H), 7.66 (d, J = 8.8 Hz, 1H), 7.57 (s, 1H), 7.42 (t, J = 5.9 Hz, 1H), 6.83 (d, J = 5.1 Hz, 1H), 5.86 (s, 1H), 4.55 (d, J = 6.0 Hz, 2H), 4.10 (dt, J = 14.2, 7.0 Hz, 1H), 3.59 (t, J = 5.4 Hz, 2H), 3.32 (s, 5H), 3.04 (s, 1H), 2.86–2.72 (m, 6H), 2.35 (d, J = 6.1 Hz, 6H), 2.13 (s, 3H), 1.26–1.09 (m, 6H), 1.03–0.93 (m, 2H), 0.63–0.51 (m, 2H).

[0147] Example 19: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-[4-(dimethylamino)quinolin-8-yl]-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzamide (Compound 19)

[0148]

[0149] MS(ESI), m / z: 552.20 [M+H] + , 574.20 [M+Na] + ; 11H NMR (500 MHz, CDCl3) δ 11.80 (s, 1H), 8.54 (d, J = 5.2 Hz, 1H), 8.23 (s, 1H), 8.08 (d, J = 8.7 Hz, 1H), 7.77 (s, 1H), 7.64 (d, J = 8.9 Hz, 2H), 7.57 (s, 1H), 6.70 (d, J = 5.4 Hz, 1H), 5.84 (s, 1H), 4.53 (d, J = 5.8 Hz, 2H), 4.09 (dq, J = 14.2, 7.2 Hz, 1H), 3.33 (dq, J = 14.1, 7.1 Hz, 1H), 3.11 (s, 6H), 2.36 (d, J = 4.4 Hz, 7H), 2.12 (s, 3H), 1.13 (t, J = 7.1 Hz, 3H), 1.01–0.94 (m, 2H), 0.62–0.53 (m, 2H).

[0150] Example 20: 5-[4-(4-Acetylpiperazin-1-yl)quinolin-8-yl]-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzamide (Compound 20)

[0151]

[0152] MS (ESI), m / z: 635.40 [M+H] + ; 1 1H NMR (500 MHz, CDCl3) δ 11.37 (s, 1H), 8.73 (d, J = 5.1

[0153] Hz, 1H), 8.28 (s, 1H), 8.07 (d, J = 8.7 Hz, 1H), 7.78–7.74 (m, 2H), 7.58 (d, J = 2.1 Hz, 1H), 7.47 (s, 1H), 6.86 (d, J = 5.1 Hz, 1H), 5.89 (s, 1H), 4.55 (d, J = 5.9 Hz, 2H), 4.11 (dq, J = 14.2, 7.2 Hz, 1H), 3.92 (d, J = 5.5 Hz, 2H), 3.79–3.77 (m, 2H), 3.32–3.26 (m, 3H), 2.37 (d, J = 11.5 Hz, 7H), 2.18 (d, J = 5.3 Hz, 6H), 1.14 (t, J = 7.1 Hz, 3H), 1.03–0.95 (m, 2H), 0.65–0.52 (m, 2H).

[0154] Example 21: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-1-isopropyl-6-[4-(4-(2-methoxyethyl)piperazin-1-yl)quinolin-7-yl]-1H-indazole-4-carboxamide (Compound 21)

[0155]

[0156] MS(ESI), m / z: 608.20 [M+H] + , 630.20 [M+Na] + ; 1 H NMR(500 MHz, CDCl3) δ 12.37(s, 1H), 8.70(d, J = 5.0 Hz, 1H), 8.45(s, 1H), 8.34(d, J = 2.0 Hz, 1H), 8.07(s, 1H), 8.04(d, J = 4.2 Hz, 1H), 7.93(s, 1H), 7.86(s, 1H), 7.81(dd, J = 8.7, 2.0 Hz, 1H), 6.84(d, J = 5.1 Hz, 1H), 5.84(s, 1H), 4.95–4.91(m, 1H), 4.65(d, J = 5.7 Hz, 2H), 3.60(t, J = 5.4 Hz, 3H), 3.35(t, J = 4.9 Hz, 4H), 2.84(s, 6H), 2.38(s, 4H), 2.11(s, 4H), 1.61(d, J = 6.6 Hz, 6H).

[0157] Example 22: 6-[4-(4-Acetylpiperazin-1-yl)quinolin-7-yl]-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-1-isopropyl-1H-indazole-4-carboxamide (Compound 22)

[0158]

[0159] MS(ESI), m / z: 592.30 [M+H] + , 614.30 [M+Na] + ; 11H NMR (500 MHz, CDCl3) δ 12.00 (s, 1H), 8.81–8.69 (m, 1H), 8.47–8.37 (m, 2H), 8.18–8.03 (m, 2H), 7.94–7.84 (m, 3H), 6.92–6.81 (m, 1H), 5.88 (d, J = 3.5 Hz, 1H), 4.96 (dq, J = 18.5, 6.1 Hz, 1H), 4.65 (d, J = 5.4 Hz, 2H), 3.98–3.76 (m, 4H), 3.38–3.23 (m, 4H), 2.40 (d, J = 3.5 Hz, 3H), 2.23–2.07 (m, 6H), 1.68–1.60 (m, 6H).

[0160] Example 23: N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(N-ethylcyclopropanecarboxamido)-5-(4-(4-methoxypiperidin-1-yl)quinolin-8-yl)-2-methylbenzamide (Compound 23)

[0161]

[0162] MS (ESI), m / z: 622.30 [M+H] + ,644.30 [M+Na] + ; 1 1H NMR (400 MHz, CDCl3) δ 11.95 (s, 1H), 8.66 (d, J = 5.4 Hz, 1H), 8.26 (s, 1H), 8.03 (d, J = 8.8 Hz, 1H), 7.77 (s, 1H), 7.70 (d, J = 8.8 Hz, 1H), 7.58 (s, 1H), 6.84 (d, J = 5.4 Hz, 1H), 5.85 (s, 1H), 4.54 (d, J = 5.9 Hz, 2H), 4.11 (h, J = 7.4 Hz, 1H), 3.60–3.50 (m, 3H), 3.33 (dq, J = 14.0, 7.1 Hz, 1H), 3.15 (t, J = 9.4 Hz, 2H), 2.36 (s, 7H), 2.15 (s, 6H), 1.93 (t, J = 10.6 Hz, 2H), 1.14 (t, J = 7.6 Hz, 6H), 1.04–0.92 (m, 2H), 0.67–0.51 (m, 2H).

[0163] Example 24: N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(N-ethylcyclopropanecarboxamido)-2-methyl-5-(4-(4-(methylsulfonyl)piperazin-1-yl)quinazolin-8-yl)benzamide (Compound 24)

[0164]

[0165] MS(ESI), m / z: 672.30 [M+H] + , 694.20 [M+Na] + ; 1 H NMR(500 MHz, CDCl3) δ 8.74 (s, 1H), 8.17 (s, 1H), 7.93 (s, 1H), 7.76 (d, J = 6.6 Hz, 2H), 7.55 (s, 1H), 5.93 (s, 1H), 4.55 (s, 2H), 4.13–4.01 (m, 4H), 3.46 (t, J = 4.8 Hz, 4H), 2.85 (s, 2H), 2.38 (d, J = 16.4 Hz, 7H), 2.23 (s, 3H), 1.14 (t, J = 7.1 Hz, 3H), 0.99 (d, J = 20.8 Hz, 2H), 0.64–0.53 (m, 2H).

[0166] Example 25: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-(N-ethylcyclopropanecarboxamido)-5-[4-(4-ethylpiperazin-1-yl)quinazolin-8-yl]-2-methylbenzamide (Compound 25)

[0167]

[0168] MS(ESI), m / z: 622.30 [M+H] + , 644.30 [M+Na] + ; 1 H NMR(500 MHz, CDCl3) δ 11.44 (s, 1H), 8.68 (d, J = 3.5 Hz, 1H), 8.05 (s, 1H), 7.93 (dd, J = 8.7, 3.7 Hz, 1H), 7.72 (s, 1H), 7.63 (d, J = 8.9 Hz, 1H), 7.55 (s, 1H), 7.48 (s, 1H), 5.90 (d, J = 3.6 Hz, 1H), 4.56 (d, J = 5.7 Hz, 2H), 4.02 (d, J = 96.1 Hz, 4H), 3.38–3.05 (m, 2H), 2.73 (s, 4H), 2.64–2.51 (m, 3H), 2.38 (dd, J = 15.4, 3.6 Hz, 6H), 2.17 (d, J = 3.7 Hz, 3H), 1.17 (dtd, J = 25.0, 7.4, 3.4 Hz, 6H), 1.06–0.93 (m, 2H), 0.66–0.53 (m, 2H).

[0169] Example 26: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-[4-((2S,6R)-2,6-dimethylmorpholino)quinazolin-8-yl]-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzamide (Compound 26)

[0170]

[0171] MS(ESI), m / z: 623.30 [M+H] + ; 1 H NMR(500 MHz, CDCl3) δ 11.33(s, 1H), 8.70(s, 1H), 8.11–8.03(m, 1H), 7.92(d, J = 8.7 Hz, 1H), 7.73(d, J = 2.1 Hz, 1H), 7.67(dd, J = 8.8, 2.0 Hz, 1H), 7.56(d, J = 2.0 Hz, 1H), 7.46(s, 1H), 5.90(s, 1H), 4.56(d, J = 5.9 Hz, 2H), 4.28(d, J = 12.9 Hz, 2H), 4.11(dq, J = 14.2, 7.2 Hz, 1H), 3.84(dt, J = 14.2, 7.8 Hz, 2H), 3.33(dq, J = 14.1, 7.1 Hz, 1H), 3.01(dd, J = 13.1, 10.4 Hz, 2H), 2.38(d, J = 14.5 Hz, 7H), 2.19(s, 3H), 1.27(d, J = 6.2 Hz, 6H), 1.14(t, J = 7.1 Hz, 3H), 1.00(dt, J = 17.8, 5.8 Hz, 2H), 0.64–0.55(m, 2H).

[0172] Example 27: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-(N-ethylcyclopropanecarboxamido)-2-methyl-5-[4-(4-methylpiperidin-1-yl)quinolin-8-yl]benzamide (Compound 27)

[0173]

[0174] MS(ESI), m / z: 606.30 [M+H] + ; 1 H NMR(400 MHz, CDCl3) δ 12.01(s, 1H), 8.64(d, J = 5.3

[0175] Hz, 1H), 8.22 (s, 1H), 8.02 (d, J = 8.8 Hz, 1H), 7.75 (s, 1H), 7.65 (d, J = 8.8 Hz, 1H), 7.54 (d, J = 23.4 Hz, 2H), 6.81 (d, J = 5.3 Hz, 1H), 5.84 (s, 1H), 4.54 (d, J = 5.9 Hz, 2H), 4.11 (dq, J = 14.2, 7.1 Hz, 1H), 3.64 (d, J = 12.5 Hz, 2H), 3.32 (dt, J = 13.6, 7.1 Hz, 1H), 2.92–2.84 (m, 2H), 2.35 (s, 7H), 2.13 (s, 3H), 1.85 (dd, J = 12.8, 3.4 Hz, 2H), 1.55 (dt, J = 12.3, 6.1 Hz, 2H), 1.21 (q, J = 3.9 Hz, 1H), 1.14 (t, J = 7.2 Hz, 3H), 1.07 (d, J = 6.4 Hz, 3H), 0.97 (dd, J = 13.0, 6.3 Hz, 2H), 0.61–0.54 (m, 2H).

[0176] Example 28: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-(N-ethylcyclopropanecarboxamido)-2-methyl-5-[4-(4-methylpiperazin-1-yl)quinazolin-8-yl]benzamide (Compound 28)

[0177]

[0178] MS(ESI), m / z: 608.30 [M + H] + ,630.30 [M + Na] + ; 1 1H NMR (500 MHz, CDCl3) δ 11.32 (s, 1H), 8.68 (s, 1H), 8.02 (s, 1H), 7.93 (s, 1H), 7.72–7.52 (m, 3H), 5.89 (s, 1H), 4.57 (s, 2H), 4.11 (dq, J = 14.2, 7.1 Hz, 1H), 3.87 (t, J = 5.0 Hz, 4H), 3.32 (dq, J = 13.9, 7.0 Hz, 1H), 2.66 (t, J = 4.9 Hz, 4H), 2.37 (d, J = 13.2 Hz, 10H), 2.16 (s, 3H), 1.14 (t, J = 7.1 Hz, 3H), 1.04–0.95 (m, 2H), 0.59 (d, J = 24.1 Hz, 2H).

[0179] Example 29: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-(N-ethylcyclopropanecarboxamido)-5-[4-(4-methoxypiperidin-1-yl)quinazolin-8-yl]-2-methylbenzamide (Compound 29)

[0180]

[0181] MS(ESI), m / z: 623.30 [M+H] + , 645.30 [M+Na] + ; 1 1H NMR (500 MHz, CDCl3) δ 11.65 (s, 1H), 8.66 (d, J = 3.5 Hz, 1H), 8.06 (s, 1H), 7.90 (dd, J = 8.8, 3.5 Hz, 1H), 7.73 (t, J = 2.7 Hz, 1H), 7.63 (d, J = 8.8 Hz, 1H), 7.59–7.47 (m, 2H), 5.89 (d, J = 3.4 Hz, 1H), 4.55 (d, J = 5.1 Hz, 2H), 4.09 (d, J = 6.5 Hz, 4H), 3.68–3.52 (m, 4H), 3.42 (d, J = 3.4 Hz, 3H), 3.38–3.27 (m, 2H), 2.36 (d, J = 3.4 Hz, 6H), 2.17 (d, J = 3.5 Hz, 3H), 2.06 (s, 3H), 1.86–1.78 (m, 2H), 1.16–1.11 (m, 3H), 1.04–0.95 (m, 2H), 0.59 (d, J = 26.5 Hz, 2H).

[0182] Example 30: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-1-isopropyl-6-[4-(4-(methylsulfonyl)piperazin-1-yl)quinazolin-7-yl]-1H-indazole-4-carboxamide (Compound 30)

[0183]

[0184] MS(ESI), m / z: 629.20 [M+H] + , 651.20 [M+Na] + ; 11H NMR (500 MHz, CDCl3) δ 12.10 (s, 1H), 8.75 (s, 1H), 8.45 (s, 1H), 8.19 (s, 1H), 8.04 (s, 1H), 7.93–7.79 (m, 4H), 5.89 (s, 1H), 4.92 (p, J = 6.7 Hz, 1H), 4.64 (d, J = 5.6 Hz, 2H), 3.92 (t, J = 5.0 Hz, 4H), 3.44 (t, J = 4.9 Hz, 4H), 2.85 (s, 3H), 2.40 (s, 3H), 2.16 (s, 3H), 1.61 (d, J = 6.6 Hz, 6H).

[0185] Example 31: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-6-[4-((2S,6R)-2,6-dimethylmorpholino)quinazolin-7-yl]-1-isopropyl-1H-indazole-4-carboxamide (Compound 31)

[0186]

[0187] MS (ESI), m / z: 580.30 [M+H] + ; 1 1H NMR (400 MHz, CDCl3) δ 11.94 (s, 1H), 8.64 (s, 1H), 8.43 (s, 1H), 8.17–7.97 (m, 3H), 7.83 (dd, J = 25.0, 12.6 Hz, 3H), 5.93 (s, 1H), 4.90 (dd, J = 14.2, 7.5 Hz, 1H), 4.66 (d, J = 19.1 Hz, 2H), 4.28 (d, J = 13.1 Hz, 2H), 3.82 (s, 2H), 3.06–2.95 (m, 2H), 2.38 (s, 3H), 2.19 (s, 3H), 1.59 (d, J = 6.6 Hz, 6H), 1.26 (d, J = 6.3 Hz, 6H).

[0188] Example 32: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-6-[4-(4-ethylpiperazin-1-yl)quinazolin-7-yl]-1-isopropyl-1H-indazole-4-carboxamide (Compound 32)

[0189]

[0190] MS (ESI), m / z: 579.30 [M+H] + , 601.30 [M+Na] + ; 11H NMR (400 MHz, CDCl3) δ 9.37 (d, J =

[0191] 26.7 Hz, 2H), 8.64 (s, 1H), 8.28 (d, J = 59.3 Hz, 3H), 7.61 (d, J = 71.2 Hz, 1H), 7.41–7.14 (m, 1H), 7.01 (s, 1H), 5.42 (s, 1H), 4.81 (s, 2H), 4.45–3.85 (m, 4H), 3.60–3.12 (m, 2H), 2.69–2.30 (m, 6H), 1.79 (d, J = 6.1 Hz, 4H), 1.27–1.19 (m, 9H).

[0192] Example 33: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-1-isopropyl-6-[4-(4-methoxypiperidin-1-yl)quinazolin-7-yl]-1H-indazole-4-carboxamide (Compound 33)

[0193]

[0194] MS (ESI), m / z: 580.30 [M+H] + ; 1 1H NMR (500 MHz, CDCl3) δ 12.26 (s, 1H), 8.67 (s, 1H), 8.46 (s, 1H), 8.17 (s, 1H), 8.08 (s, 1H), 7.90 (d, J = 5.5 Hz, 2H), 7.83 (s, 1H), 7.78 (d, J = 8.6 Hz, 1H), 5.88 (s, 1H), 4.92 (p, J = 6.7 Hz, 1H), 4.65 (d, J = 5.7 Hz, 2H), 4.07 (s, 2H), 3.63–3.55 (m, 3H), 3.42 (s, 3H), 2.39 (s, 3H), 2.14 (s, 3H), 2.07 (s, 2H), 1.86–1.80 (m, 2H), 1.61 (d, J = 6.6 Hz, 6H).

[0195] Example 34: N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-1-isopropyl-6-[4-(4-methylpiperazin-1-yl)quinazolin-7-yl]-1H-indazole-4-carboxamide (Compound 34)

[0196]

[0197] MS (ESI), m / z: 565.30 [M+H] + , 587.20 [M+Na]+ ; 1 1H NMR (500 MHz, CDCl3) δ 12.47 (s, 1H), 8.72 (s, 1H), 8.45 (s, 1H), 8.15 (d, J = 2.0 Hz, 1H), 8.03 (t, J = 5.8 Hz, 1H), 7.94–7.90 (m, 2H), 7.83 (s, 1H), 7.76 (dd, J = 8.7, 1.9 Hz, 1H), 5.87 (s, 1H), 4.92 (p, J = 6.7 Hz, 1H), 4.66 (d, J = 5.8 Hz, 2H), 3.88 (d, J = 5.1 Hz, 4H), 2.67 (t, J = 4.9 Hz, 4H), 2.41 (d, J = 10.5 Hz, 6H), 2.13 (s, 3H), 1.61 (d, J = 6.6 Hz, 6H).

[0198] Example 35: Antitumor cell activity in vitro

[0199] The in vitro inhibitory activity of novel compounds containing a biphenylamide analogue structure against human B lymphoma cells (SU-DHL-6, WSU-DLCL-2) was screened.

[0200] 1) Cell seeding: Take the SU-DHL-6 cell suspension and WSU-DLCL-2 cell suspension in the logarithmic growth phase for centrifugation (800 r, 5 min). After discarding the supernatant, add 1 mL of fresh medium to resuspend the cells. Take 100 μL of the cell suspension and mix it with 900 μL of medium to prepare a 1:10 dilution. Use a hemocytometer for cell quantification. Adjust the concentration according to the counting results, and inoculate 100 μL of the suspension containing 1×10 4 cells into each well of a 96-well plate. After inoculation, place the culture plate in a 37 °C, 5% CO2 incubator for pre-culture for 24 h.

[0201] 2) Drug preparation: Accurately weigh the test compound and the positive control drug Tazematostat using an analytical balance. Initially, dissolve the sample in 100 μL of DMSO, add 50 μL of Tween 80 to assist dissolution, and then make up the volume to 2 mL with RPMI-1640 medium. Obtain a clear stock solution (1 mmol / L) by vortex oscillation. All solution preparation processes need to be completed under sterile conditions.

[0202] 3) Gradient drug addition: Take the stock solution for serial gradient dilution to obtain five concentration gradients of the drug solution: 100 μmol / L, 10 μmol / L, 1 μmol / L, 0.1 μmol / L, and 0.01 μmol / L. Add 100 μL of the corresponding concentration of the drug solution to each well of the pre-cultured 96-well plate (the actual concentration in the well is 50% of the labeled concentration at this time). After drug addition, continue the culture for 72 h for efficacy observation.

[0203] 4) Detection by MTT method: Add 20 μL of 0.5% MTT solution (final concentration 0.1%) to each well under light protection conditions. After incubation at 37 °C for 4 h, centrifuge (1500 rpm, 5 min) to remove the supernatant. Add 150 μL of DMSO to each well and mix thoroughly by oscillation until the formazan crystals are completely dissolved. Measure the absorbance at a wavelength of 490 nm using an enzyme-linked immunosorbent assay (ELISA) reader. Draw a dose-effect curve by calculating the cell survival rate of different concentration groups, and calculate the IC 50 value. The results are shown in Table 1.

[0204] Table 1 Results of in vitro anti-tumor cell activity of compounds

[0205]

[0206] Example 36: EZH2 kinase inhibition experiment

[0207] 1) Reagent preparation: Prepare a reaction buffer system, an enzyme working solution, a substrate working solution, and a radioactive-labeled [3H]-S-adenosylmethionine ([3H]-SAM) solution in advance.

[0208] 2) Enzymatic reaction: Dispense 10 μL of the enzyme reaction solution into each well of a 96-well ELISA plate and incubate at room temperature for 15 min. Sequentially add 10 μL of the substrate working solution and 10 μL of the [3H]-SAM solution to each reaction well to initiate the reaction process. Then place the reaction system in a constant temperature environment at 25 °C for 90 min. Add 5 μL of the termination buffer to terminate the reaction, and then quantitatively transfer 25 μL of the reaction mixture to a Flashplate using a pipette.

[0209] 3) Result analysis: Collect radioactive signals from the Flashplate using a liquid scintillation counter, and calculate the inhibition rate according to the formula:

[0210] Inhibition rate (%) = (Count of blank group - Count of experimental group) × 100%

[0211] Evaluate the inhibitory effect of enzyme activity by comparing the differences in radioactive intensities of different treatment groups.

[0212] The results of the inhibition of EZH2 kinase activity by the compound at concentrations of 10000 nM, 100 nM, and 10 nM are shown in Table 2. ND indicates not tested.

[0213] Table 2 Results of compound kinase inhibition experiment

[0214]

[0215]

[0216] Example 37: In vitro kinase activity assay (IC50)

[0217] The in vitro EZH2 kinase activity of novel compounds containing the biphenylamide analogue structure was further assayed, and the results are shown in Table 3.

[0218] Table 3 In vitro EZH2 kinase activity (IC 50 ) results

[0219] Compd <![CDATA[IC 50 (nM)]]> Compound 15 0.375 Compound 22 21.00 Compound 24 1.31 Compound 25 1.04 Compound 27 1.10 Compound 28 0.95

[0220] It can be clearly seen from the above test results that the novel compounds containing the biphenylamide analogue structure of general formula (I) or (II) to be protected by the present invention have good in vitro anti-tumor activity and kinase inhibitory activity. Such compounds have good prospects for development and application as anti-tumor drugs.

[0221] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various equivalent transformations can be made to the technical solutions of the present invention. To avoid unnecessary repetition, the present invention will not separately describe various possible combinations. Any modification, equivalent replacement or improvement made within the scope of the technical concept of the present invention is included in the protection scope of the present invention.

Claims

1. A novel compound containing a biphenylamide analogue structure and its pharmaceutically acceptable salts, characterized in that, Having the structural general formula shown in (Ⅰ) or (Ⅱ): Wherein, A is selected from primary amine compounds; B is selected from isopropyl or cyclopentyl; X is selected from C or N atoms; R is selected from secondary amines or structures containing secondary amine substitutions.

2. The novel compound containing a biphenylamide analog structure and its pharmaceutically acceptable salt according to claim 1, characterized in that A is selected from 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one, 3-aminopiperidine-2,6-dione, 2-(3-indolyl)ethylamine or 2-(1H-imidazol-4-yl)ethylamine; R is selected from (2S,6R)-2,6-dimethylmorpholine, 1-methylpiperazine, 4-methoxypiperidine, 4-methylpiperidine, 1-(2-methoxyethyl)piperazine, 1-(piperazin-1-yl)ethanone, 3-isopropoxyazetidine, morpholine, 1-ethylpiperazine, 1-methanesulfonylpiperazine or dimethylamine.

3. The novel compound containing a biphenylamide analogue structure according to claim 2 and its pharmaceutically acceptable salts, characterized in that, The novel compound containing a biphenylamide analog structure has the following structural formula:

4. A pharmaceutical composition, characterized in that, Comprising the novel compound containing a biphenylamide analog structure according to any one of claims 1-3 and its pharmaceutically acceptable salt as an active ingredient combined with a pharmaceutically acceptable carrier.

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

6. The pharmaceutical composition according to claim 4, wherein The pharmaceutical composition is made into dosage forms such as tablets, capsules, granules, sprays or injections.

7. Use of the novel compound containing a biphenylamide analog structure according to any one of claims 1-3 and its pharmaceutically acceptable salt or the pharmaceutical composition according to any one of claims 4-6 in the preparation of a drug for treating diseases mediated by EZH2, PRC2 or EZH2 / PRC2.

8. The application according to claim 7, wherein Use of the novel compound containing a biphenylamide analog structure according to any one of claims 1-3 and its pharmaceutically acceptable salt or the pharmaceutical composition according to any one of claims 4-6 in the preparation of a drug with EZH2 as a molecular therapeutic target.

9. Use of the novel compound containing a biphenylamide analog structure according to any one of claims 1-3 and its pharmaceutically acceptable salt or the pharmaceutical composition according to any one of claims 4-6 in the preparation of an anti-tumor drug.

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

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