Benzamide compounds containing imidazolinone structure and preparation method and application thereof

Abstract

Benzamide compounds containing imidazolinone structure and preparation methods and applications thereof belong to the field of medicine. The present invention relates to benzamide compounds containing imidazolinone structure as shown in general formula I or pharmaceutically acceptable salts thereof, wherein the substituent R 1 , R 2 , R 3 , R 4 and R 5 The meanings of are explained in the present specification. The compound of formula I or a pharmaceutically acceptable salt thereof involved in the present invention has a significant inhibitory effect on wild-type and mutant EZH2, and can be used to prepare drugs for inhibiting EZH2, especially drugs for preventing and / or treating tumors.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and specifically relates to a class of benzamide compounds containing an imidazolidinone structure, and a preparation method and application thereof. Background Art

[0002] Polycomb repressive complex 2, or PRC2, is a multiprotein complex that catalyzes the methylation of lysine 27 (H3K27) of histone H3, thereby silencing related genes. PRC2 consists of four core parts: EZH1 or EZH2 (SET domain with histone methyltransferase activity), EED, SUZ12, and RbAp48 (histone binding domain). EZH2 is the core catalytic subunit of PRC2, which needs to interact with EED and then rely on the stabilization of SUZ12 to exert methyltransferase activity. EZH2 is often overexpressed in prostate, breast, kidney, lung, myeloma, and lymphoma. Wild-type EZH2 shows the best catalytic efficiency for monomethylation of unmethylated H3K27 sites, and its catalytic efficiency decreases with the increase of the methylation state of lysine amino acids. In addition, Y641 mutants within the EZH2 catalytic domain have been identified in non-Hodgkin's lymphomas such as diffuse large B-cell lymphoma and follicular lymphoma, and the disease-associated EZH2 Y641 mutant can effectively catalyze dimethylation and trimethylation reactions. Therefore, wild-type EZH2 and mutant EZH2 will synergistically lead to increased H3K27Me3 levels, resulting in silencing of target genes that promote normal cell differentiation and inhibit proliferation. Therefore, it is meaningful to develop EZH2 inhibitors that are highly active against both wild-type and mutant forms for the treatment of non-Hodgkin's lymphomas such as DLBCL.

[0003] After several years of efforts, several EZH2 inhibitors have entered the clinical research stage, including Tazemetostat, GSK126, CPI1209, CPI0209, SHR2554, PF06821497, and DS3201b. Among them, Tazemetostat has been approved by the FDA for the treatment of adult patients with metastatic / locally advanced epithelioid sarcoma that cannot be completely removed and pediatric patients over 16 years old. It is the first FDA-approved drug for the treatment of epithelioid sarcoma. It was later approved for the treatment of follicular lymphoma and is also the first approved EZH2 inhibitor. However, for the indication of DLBCL, Tazemetostat combined with R-CHOP chemotherapy is still in the clinical research stage. These inhibitors require further clinical trials to evaluate their potential application in the treatment of DLBCL. Therefore, it is necessary to develop novel EZH2 inhibitors to address the limitations of clinical treatment of DLBCL. Summary of the invention

[0004] The present invention provides a series of novel benzamide compounds containing imidazolinone structures. Studies have shown that the compounds can significantly inhibit the enzyme activities of EZH2 WT and EZH2 mutants. The present invention relates to benzamide compounds containing imidazolinone structures as shown in general formula I or pharmaceutically acceptable salts thereof, their preparation methods and pharmaceutical compositions containing the compounds, wherein the substituent R 1 , R 2 , R 3 , R 4 , R 5 The meanings of are explained in the specification. The present invention relates to compounds having high levels of inhibitory activity against EZH2WT and EZH2 mutant enzymes, and the benzamide compounds containing imidazolinone structures or pharmaceutically acceptable salts thereof are used to prepare drugs for treating diseases associated with abnormal expression of EZH2, such as cancer and autoimmune diseases.

[0005] The present invention relates to a benzamide compound containing an imidazolidinone structure as shown in general formula I or a pharmaceutically acceptable salt thereof.

[0006]

[0007] in

[0008] R 1 is (C1-C3)alkyl or (C1-C3)alkoxy;

[0009] R 2 , R 3 Each is independently selected from hydrogen, (C1-C4) alkyl, (C3-C8) cycloalkyl, (C3-C6) cycloalkylacyl, 4-7 membered heterocycloalkyl containing 1-2 atoms selected from N, O and S, wherein the (C3-C8) cycloalkyl may be optionally replaced by 1-3 R 6 When the 4-7 membered heterocycloalkyl group containing 1-2 atoms selected from N, O and S is a heterocycloalkyl group containing sulfur atoms, the sulfur atoms therein may be oxidized to form sulfones; when the 4-7 membered heterocycloalkyl group containing 1-2 atoms selected from N, O and S is a heterocycloalkyl group containing nitrogen atoms, the nitrogen atoms therein may be optionally replaced by 1-2 R 7 replace;

[0010] R 4 , R 5Each is independently selected from hydrogen, (C1-C4) alkyl, (C1-C4) alkoxy, (C3-C8) cycloalkyl, 4-7 membered heterocycloalkyl containing 1-2 atoms selected from N, O and S, and a six- to eight-membered bicyclic ring containing 1-2 nitrogen atoms, wherein the (C1-C4) alkyl, (C1-C4) alkoxy, (C3-C8) cycloalkyl may be optionally replaced by 1-3 R 6 When the 4-7 membered heterocycloalkyl containing 1-2 atoms selected from N, O and S is a heterocycloalkyl containing sulfur atoms, the sulfur atoms therein may be oxidized to form sulfones; when the 4-7 membered heterocycloalkyl containing 1-2 atoms selected from N, O and S and the six to eight membered bicyclic ring containing 1-2 nitrogen atoms are heterocycloalkyl containing nitrogen atoms, the nitrogen atoms therein may be optionally replaced by 1-2 R 7 replace;

[0011] R 6 is selected from hydrogen, halogen, hydroxy, cyano, (C1-C3) alkyl, (C1-C3) alkoxy, (C3-C6) cycloalkyl, optionally substituted by 1-2 R 7 Substituted amino groups;

[0012] R 7 Selected from hydrogen, halogen, hydroxy, (C1-C4) alkyl, (C1-C4) alkoxy, hydroxy (C1-C4) alkyl, (C4-C6) cycloalkyl, (C1-C4) acyl, (C3-C6) cycloalkyl acyl, (C1-C4) sulfonyl, cyclopropylsulfonyl, (C1-C4) alkyl substituted by 1-3 cyano or fluorine atoms.

[0013] The present invention preferably relates to a benzamide compound containing an imidazolidinone structure of general formula I or a pharmaceutically acceptable salt thereof, wherein:

[0014] R 1 is methyl or methoxy;

[0015] Selected from:

[0016]

[0017] Selected from:

[0018]

[0019]

[0020] The benzamide compounds containing an imidazolidinone structure of the general formula I of the present invention or their pharmaceutically acceptable salts are ultimately preferably selected from compounds (I-1 to I-75), but these compounds do not imply any limitation on the present invention:

[0021]

[0022]

[0023]

[0024]

[0025] The benzamide compound containing an imidazolinone structure of the general formula I described above or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt includes a salt formed with an inorganic acid and an organic acid; the inorganic acid is selected from the group consisting of hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, and phosphoric acid; the organic acid is selected from the group consisting of succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, and p-toluenesulfonic acid;

[0026] In the present invention, "halogen" refers to fluorine, chlorine, bromine or iodine; "alkyl" refers to a straight chain or branched chain alkyl; "heterocycloalkyl" refers to a monocyclic system containing N, O and / or S atoms; Represents the place where substituents are attached.

[0027] The present invention can contain the benzamide compound containing the imidazolinone structure of the above-mentioned general formula I or its pharmaceutically acceptable salt as an active ingredient, and mix it with a pharmaceutically acceptable carrier or excipient to prepare a composition. The carrier or excipient includes a diluent, adhesive, wetting agent, disintegrant, lubricant, and glidant known in the art. Diluents include starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, and calcium hydrogen phosphate; wetting agents include water, ethanol, isopropanol, etc.; adhesives include starch slurry, dextrin, syrup, honey, glucose solution, acacia gum slurry, gelatin slurry, sodium carboxymethyl cellulose, hydroxypropyl methylcellulose, ethyl cellulose, and polyethylene glycol; disintegrants include dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, and sodium dodecyl sulfate; lubricants and glidants include talc, silicon dioxide, and polyethylene glycol.

[0028] The pharmaceutical composition of the present invention can be formulated into several dosage forms, including injections, tablets, and capsules.

[0029] The benzamide compound containing an imidazolidinone structure or a pharmaceutically acceptable salt thereof of the present invention can be used in combination with other active ingredients to achieve a better therapeutic effect.

[0030] The present invention also provides the use of a benzamide compound containing an imidazolinone structure of general formula I or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and / or treating a disease related to abnormal expression of EZH2. The disease related to abnormal expression of EZH2 is cancer and autoimmune disease. The cancer is lymphoma, non-small cell lung cancer, small cell lung cancer, head and neck cell cancer, bladder cancer, gastric cancer, colon cancer, colorectal cancer, kidney cancer, bile duct cancer, gastric cancer, esophageal squamous cell carcinoma, ovarian cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, brain cancer, melanoma, multiple myeloma, glioblastoma, leukemia or cervical cancer; the autoimmune disease is inflammatory bowel inflammation, autoimmune encephalomyelitis or multiple sclerosis.

[0031] The positive and progressive effects of the present invention are that the benzamide compounds containing imidazolinone structures of the present invention have novel chemical structures, and most of the compounds have significant inhibitory activity against EZH2 WT and EZH2 mutants in in vitro biological activity studies, and have strong anti-proliferative activity against Karpas-422 cells. The compounds can be used for the treatment and prevention of various diseases such as cancer. DETAILED DESCRIPTION

[0032] Route 1:

[0033]

[0034] (a) 4-bromo-1-fluoro-2-nitrobenzene is used as a raw material and reacts with aliphatic amines to obtain intermediate 2 through nucleophilic substitution reaction;

[0035] (b) using intermediate 2 as a raw material, obtaining intermediate 3 by nitro reduction reaction;

[0036] (c) Intermediate 3 was used as the starting material to obtain intermediate 4 through a CDI-mediated cyclization reaction;

[0037] (d) intermediate 5 is used as a raw material, condensed with tetrahydropyrone and reacted with sodium triacetoxyborohydride to obtain intermediate 6;

[0038] (e) intermediate 6 is used as a raw material, condensed with acetaldehyde and reacted with sodium triacetoxyborohydride to obtain intermediate 7;

[0039] (f) intermediate 7 is used as a raw material, and intermediate 8 is prepared by Miyaura borate esterification reaction with diboric acid pinacol ester;

[0040] (g) intermediate 8 is used as a raw material, and intermediate 4 is reacted with intermediate 9 through Suzuki-Miyaura coupling reaction;

[0041] (h) intermediate 9 is used as a raw material, and reacts with iodomethane or iodoethane to obtain intermediate 10 through nucleophilic substitution reaction;

[0042] (i) Using intermediate 10 as a raw material, intermediate 11 is obtained by ester hydrolysis under the action of NaOH;

[0043] (j) Intermediate 11 is used as a raw material, and 3-(aminomethyl)-4,6-dimethyl-1H-pyridin-2-one hydrochloride or 3-(aminomethyl)-4-methoxy-6-methylpyridin-2(1H)-one hydrochloride is reacted with a condensing agent to obtain the target compound of formula I through an amide condensation reaction.

[0044] Route 2:

[0045]

[0046] Referring to (g), (h), (i) and (j) in route 1, intermediate 12 is reacted through four steps (a), (b), (c) and (d) to obtain the target compound of formula I.

[0047] Route 3:

[0048]

[0049] Wherein, ring A is an aliphatic heterocycle containing a nitrogen atom, as defined in general formula I.

[0050] Similarly, referring to the reaction of route 1, intermediate 14 is reacted via four steps (a), (b), (c) and (d) to obtain intermediate 18.

[0051] (e) Using intermediate 18 as a starting material, removing the Boc protecting group under acidic conditions to obtain intermediate 19;

[0052] (f) Intermediate 19 is used as a raw material to react with an acyl chloride or sulfonyl chloride through N-acylation or N-sulfonylation reaction, or condense with a fatty aldehyde or ketone and react with sodium triacetoxyborohydride to obtain the target compound of formula I.

[0053] Route 4:

[0054]

[0055] Similarly, referring to the reaction of Scheme 4, intermediate 20 is reacted through six steps (a), (b), (c), (d), (e) and (f) to obtain the target compound of Formula I.

[0056] Route 5:

[0057]

[0058] (a) Intermediate 14 is used as a raw material, and reacts with iodomethane or iodoethane to obtain intermediate 26 through nucleophilic substitution reaction;

[0059] (b) Using intermediate 26 as the starting material, the Boc protecting group was removed under acidic conditions to obtain intermediate 27;

[0060] (c) intermediate 27 is used as a raw material, condensed with formaldehyde solution and reacted with sodium triacetoxyborohydride to obtain intermediate 28;

[0061] Similarly, referring to the reaction in Scheme 3, intermediate 28 is reacted in five steps (d), (e), (f), (g) and (h) to obtain the target compound of Formula I.

[0062] Route 6:

[0063]

[0064] Wherein, ring B is a cycloalkane, as defined in formula I.

[0065] Similarly, referring to the reaction in Scheme 5, intermediate 34 is reacted in five steps (a), (b), (c), (d) and (e) to obtain the target compound of Formula I.

[0066] Route 7:

[0067]

[0068] (a) Intermediate 5 is used as a raw material, condensed with acetaldehyde and reacted with sodium cyanoborohydride to obtain intermediate 39;

[0069] (b) intermediate 39 is used as a raw material, and reacted with cycloalkylcarbonyl chloride to obtain intermediate 7 through N-acylation reaction;

[0070] Similarly, referring to the reaction in Scheme 6, intermediate 7 is reacted in four steps (c), (d), (e) and (f) to obtain the target compound of Formula I.

[0071] Example 1: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-1)

[0072] Step 1: 5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoic acid methyl ester

[0073] Dissolve 3-amino-5-bromo-2-methylbenzoic acid methyl ester (10 g, 40.97 mmol) in DCM (150 mL), add tetrahydropyrone (12.3 g, 122.91 mmol) and acetic acid (7.35 g, 122.91 mmol) in turn, stir at room temperature for 1 h, then add sodium triacetoxyborohydride (26.05 g, 122.91 mmol), stir at room temperature for 3 h. TLC monitors the reaction to be complete. The reaction solution is poured into water, extracted with dichloromethane 3 times, the organic phases are combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated to obtain a yellow solid (13 g, 39.76 mmol), which is directly put into the next step without purification, with a yield of 98.0%. ESI-MS m / z: 328.23 [M+H] + .

[0074] Step 2: Methyl 5-bromo-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzoate

[0075] The compound obtained in step 1 (13 g, 39.76 mmol) was dissolved in DCM (150 mL), and acetaldehyde (3.5 g, 79.51 mmol) and acetic acid (14.32 g, 238.53 mmol) were added in sequence, stirred at room temperature for 1 h, and sodium triacetoxyborohydride (25.28 g, 119.27 mmol) was added, and stirred at room temperature for 6 h. TLC monitored the reaction to be complete. The reaction solution was poured into water, extracted with dichloromethane 3 times, the organic phases were combined, washed with saturated sodium chloride, dried over anhydrous ammonium sulfate, and concentrated, and purified by column chromatography to obtain a light yellow solid (11.46 g, 32.17 mmol), with a yield of 81.0%. ESI-MS m / z: 357.25 [M+H] + .

[0076] Step 3: 3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid methyl ester

[0077] The compound obtained in step 2 (5 g, 14.03 mmol) was dissolved in 1,4-dioxane (75 mL), and diboric acid pinacol ester (5.35 g, 21.05 mmol), Pd(dppf)Cl2 (0.51 g, 0.70 mmol) and KOAc (4.13 g, 42.10 mmol) were added in sequence, and then heated to 100 ° C and refluxed for 3 h under N2 protection. TLC monitored the reaction to be complete. Cool to room temperature, add 80 mL of water, filter with diatomaceous earth, extract the filtrate with EA 3 times, combine the organic phases, wash twice with saturated sodium chloride, dry over anhydrous sodium sulfate and concentrate, and purify by column chromatography to obtain a light yellow solid (5.39 g, 13.36 mmol), with a yield of 95.2%. ESI-MS m / z: 404.32 [M+H] + .

[0078] Step 4: N-(4-bromo-2-nitrophenyl)tetrahydro-2H-pyran-4-amine

[0079] Dissolve 4-bromo-1-fluoro-2-nitrobenzene (10 g, 45.45 mmol) in DMF (100 mL), add 4-aminotetrahydropyran (6.90 g, 68.18 mmol), potassium carbonate (12.56 g, 90.91 mmol) in sequence. Heat to 60 ° C and react for 3 h. TLC monitors the reaction to be complete. Add 600 mL of water, stir for 30 min, and filter to obtain a yellow solid (13.41 g, 44.55 mmol), yield: 98.0%. ESI-MS m / z: 301.12 [M+H] + .

[0080] Step 5: 4-Bromo-N 1 -(Tetrahydro-2H-pyran-4-yl)benzene-1,2-diamine

[0081] Dissolve NH4Cl (14.30 g, 267.27 mmol) in 67 mL of water, and add iron powder (14.97 g, 267.27 mmol) in batches. Heat to 80 °C to activate the iron powder for 30 min. Then add the compound obtained in step 4 (13.41 g, 44.55 mmol) and EtOH (201 mL) in sequence, and continue the reaction for 1 h. TLC monitors the reaction to be complete. Add ethyl acetate 100 mL and water 100 mL, filter with diatomaceous earth, add EA to extract 3 times, combine the organic layers, wash with saturated sodium chloride 2 times, dry and evaporate to obtain a purple solid (9.08 g, 33.49 mmol). Yield: 75.2%. ESI-MS m / z: 271.12 [M+H] + .

[0082] Step 6: 5-Bromo-1-(tetrahydro-2H-pyran-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one

[0083] The compound obtained in step 5 (9.08 g, 33.49 mmol) was dissolved in redistilled dioxane (136 mL), and CDI (16.29 g, 100.46 mmol) was added in batches. The reaction was allowed to react at room temperature for 3 h. TLC monitored the reaction to be complete. Evaporate to dryness, add water, filter and dry, and slurry with ether to obtain a gray solid (7.78 g, 26.19 mmol), with a yield of 78.2%. ESI-MS m / z: 297.08 [M+H] + .

[0084] Step 7: Methyl 3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzoate

[0085] The compound obtained in step 3 (0.71 g, 1.77 mmol) was dissolved in 1,4-dioxane (6 mL), and the compound obtained in step 6 (0.71 g, 1.77 mmol), Pd(dppf)Cl2 (0.12 g, 0.17 mmol), K2CO3 (0.70 g, 5.05 mmol) and H2O (2 mL) were added in sequence. Under N2 protection, the mixture was heated to 100 °C for 3 h. The reaction was monitored by TLC to be complete. After cooling to room temperature, 20 mL of water was added, the mixture was filtered with diatomaceous earth pad, and the filtrate was extracted with EA 3 times. The organic phases were combined, washed with saturated sodium chloride 2 times, dried over anhydrous sodium sulfate, and concentrated. The mixture was purified by column chromatography to obtain a light yellow solid (0.51 g, 1.04 mmol) with a yield of 62.1%. ESI-MS m / z: 494.35 [M+H] + .

[0086] Step 8: 3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzoic acid methyl ester

[0087] The compound obtained in step 7 (0.51 g, 1.04 mmol) was dissolved in DMF (5 mL), and NaH (0.083 g, 2.08 mmol) was added in batches at 0°C, and stirred at room temperature for 30 min. Methyl iodide (0.29 g, 2.08 mmol) was added, and the reaction was continued at room temperature for 2 h. TLC monitored the reaction to be complete. 50 mL of water was added, and after stirring for 10 min, EA was added to extract 3 times, the organic phases were combined, washed with water twice, washed with saturated sodium chloride twice, dried over anhydrous sodium sulfate, and evaporated to obtain a light yellow solid (0.49 g, 0.96 mmol), with a yield of 92.6%. ESI-MS m / z: 508.23 [M+H] + .

[0088] Step 9: 3-(Ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzoic acid

[0089] The compound obtained in step eight (0.49 g, 0.96 mmol) was dissolved in methanol (8 mL), and NaOH (0.15 g, 3.84 mmol) and water (2 mL) were added in sequence. The temperature was raised to 66 ° C and the reaction was continued for 3 h. TLC monitored the reaction to be complete. After cooling to room temperature, 10 mL of water was added, the pH was adjusted to 4 with 2N HCl aqueous solution, and then DCM was added to extract 3 times. The organic phases were combined, washed with saturated sodium chloride 2 times, dried over anhydrous sodium sulfate, and evaporated to obtain a light yellow solid (0.45 g, 0.91 mmol), with a yield of 95.2%. ESI-MS m / z: 494.30 [M+H] + .

[0090] Step 10: Preparation of Compound I-1

[0091] The compound obtained in step nine (0.15 g, 0.30 mmol) was dissolved in DMF (2 mL), and 3-(aminomethyl)-4,6-dimethyl-1H-pyridin-2-one hydrochloride (0.057 g, 0.30 mmol), HATU (0.15 g, 0.40 mmol), and DIPEA (0.20 g, 1.52 mmol) were added in sequence, and the reaction was carried out at room temperature for 1 h. TLC monitored the reaction to be complete. 20 mL of water was added, and after stirring for 10 min, EA was added to extract 3 times, the organic phases were combined, washed with water twice, washed with saturated sodium chloride twice, dried over anhydrous sodium sulfate, evaporated to dryness, and column chromatography was performed to obtain the target compound I-1 (0.088 g, 0.14 mmol), with a yield of 45.2%. ESI-MS m / z: 628.25 [M+H] + ; 1H NMR (600MHz, DMSO-d6) δ11.46(s,1H),8.18(t,J=4.5Hz,1H),7.40(d,J=7.2Hz,2H),7.36(d,J=8.2Hz,1H),7.30 (d,J=7.9Hz,1H),7.27(s,1H),5.86(s,1H),4.49–4.44(m,1H),4.30(d,J=4.6Hz,2H),4.02–3.98(m,2H),3.86–3 .82(m,2H),3.49(t,J=11.4Hz,2H),3.39(s,3H),3.25(t,J=11.3Hz,2H),3.13–3.08(m,2H),3.05–2.99(m,1H),2 .42–2.36(m,2H),2.24(s,3H),2.22(s,3H),2.11(s,3H),1.67(s,4H),1.57–1.51(m,2H),0.85(t,J=6.8Hz,3H).

[0092] According to the synthesis method of Example 1, the target compounds in Examples 2-4, 43, 71 and 72 were prepared.

[0093] Example 2: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-1-(oxetane-3-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-2)

[0094] ESI-MS m / z:600.22[M+H] + .

[0095] Example 3: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(1-(2-methoxyethyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-3)

[0096] ESI-MS m / z:602.23[M+H] + .

[0097] Example 4: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro 2H-pyran-4-yl)amino)-5-(1-ethyl-2-oxo-3-(tetrahydro 2H-pyran-4-yl)-2,3-dihydro 1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-4)

[0098] ESI-MS m / z:642.10[M+H] + .

[0099] Example 5: 5-(1-(1-acetylpiperidin-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-5)

[0100] Step 1: tert-butyl 4-(5-(3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylphenyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-1-carboxylate

[0101] According to the synthesis method of the target compound I-1, 4-aminopiperidine-1-carboxylic acid tert-butyl ester was used to replace 4-aminotetrahydropyran to prepare a light yellow solid. ESI-MS m / z: 593.23 [M+H] + .

[0102] Step 2: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro 2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-2-oxo-1-(piperidin-4-yl)-2,3-dihydro 1H-benzo[d]imidazol-5-yl)benzamide

[0103]

[0104] The compound obtained in step 1 of Example 5 (1.64 g, 2.25 mmol) was dissolved in DCM (16 mL) and TFA (8 mL) was added. The reaction was allowed to react at room temperature for 1 h. TLC monitored the reaction to be complete. 30 mL of water was added, the pH was adjusted to 10 with 2N NaOH aqueous solution, and the mixture was extracted with DCM for 3 times. The organic phases were combined, washed with saturated sodium chloride for 2 times, dried over anhydrous sodium sulfate, and evaporated to obtain a white solid (1.34 g, 2.14 mmol). The yield was 95.2%. ESI-MS m / z: 627.35 [M+H]+ .

[0105] Step 3: Preparation of compound I-5

[0106] The compound obtained in step 2 of Example 5 (0.1 g, 0.16 mmol) and NEt3 (0.049 g, 0.48 mmol) were dissolved in THF (2 mL). A THF solution of acetyl chloride (0.015 g, 0.192 mmol) was slowly added dropwise at 0°C. The reaction was allowed to react at room temperature for 30 min. TLC monitored the reaction to be complete. Water (20 mL) was added, and the mixture was extracted with DCM for 3 times. The organic phases were combined, washed with saturated sodium chloride for 2 times, dried over anhydrous sodium sulfate, evaporated to dryness, and subjected to column chromatography to obtain a white solid with a yield of 35.2%. 1 H NMR (600MHz, DMSO-d6) δ11.49(s,1H),8.19(s,1H),7.42–7.39(m,2H),7.36(d,J=7.9Hz,1H),7.30–7.26(m,2H),5.86(s,1H),4.57(d,J=11.6Hz,1 H),4.50–4.46(m,1H),4.31(d,2H),3.97(d,J=12.2Hz,1H),3.84(d,J=9. 0Hz,2H),3.39(s,3H),3.26(d,J=11.1Hz,2H),3.20(d,J=12.7Hz,1H),3. 12–3.08(m,2H),3.05–3.01(m,1H),2.66(t,J=12.2Hz,1H),2.34–2.30(m ,1H),2.24(s,3H),2.22(s,3H),2.11(s,3H),2.07(s,3H),1.76(d,J=10. 6Hz,1H),1.72(d,J=12.5Hz,1H),1.67(d,J=11.3Hz,2H),1.56–1.51(m,2 H),1.09(t,J=6.7Hz,1H),0.85(t,J=5.9Hz,3H); ESI-MSm / z:669.25[M+H] + .

[0107] According to the synthesis method of Example 5, the target compounds in Examples 6-11, 19-21, 27, 44-46 and 53-62 were prepared.

[0108] Example 6: 5-(1-(1-(cyclopropanecarbonyl)piperidin-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-6)

[0109] ESI-MS m / z:695.38[M+H] + .

[0110] Example 7: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(3-methyl-2-oxo-1-(1-propionylpiperidin-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-7)

[0111] ESI-MS m / z:683.25[M+H] + .

[0112] Example 8: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(1-(1-isobutyrylpiperidin-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-8)

[0113] ESI-MS m / z:697.30[M+H] + .

[0114] Example 9: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-1-(1-(methylsulfonyl)piperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-9)

[0115] ESI-MS m / z:705.32[M+H] + .

[0116] Example 10: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(1-(1-(ethylsulfonyl)piperidin-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-10)

[0117] ESI-MS m / z:719.32[M+H] + .

[0118] Example 11: (1-(1-(cyclopropylsulfonyl)piperidin-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-11)

[0119] ESI-MS m / z:731.25[M+H] + .

[0120] Example 12: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-12)

[0121] The compound obtained in step 2 of Example 5 (0.1 g, 0.16 mmol) was dissolved in DCM (2 mL), and 40% formaldehyde aqueous solution (0.036 g, 0.48 mmol) and acetic acid (0.029 g, 0.48 mmol) were added in sequence, and stirred at room temperature for 30 min. Sodium triacetoxyborohydride (STAB) (0.10 g, 0.48 mmol) was then added in batches, and the reaction was allowed to proceed at room temperature for 1 h. TLC monitored the reaction to be complete. Water was added, and the pH was adjusted to 10 with 2NNaOH, and the mixture was extracted with DCM for 3 times. The organic layers were combined, washed with saturated sodium chloride for 2 times, dried over anhydrous sodium sulfate, evaporated to dryness, and column chromatographed to obtain a white solid (0.075 g, 0.12 mmol), with a yield of 72.6%. 1H NMR (600MHz, DMSO-d6) δ11.46(s,1H),8.18(t,J=4.8Hz,1H),7.40(s,1H),7.39(s,1H),7.33(d,J=8.2Hz,1H),7.30–7.28(m, 1H),7.26(s,1H),5.86(s,1H),4.30(d,J=4.8Hz,2H),4.19(ddd,J=12.4,7.2,3.5Hz,1H),3.84(d,J=9.9Hz,2H),3.38(s,3H) ,3.25(t,J=11.2Hz,2H),3.10(q,J=6.5Hz,2H),3.04–3.00(m,1H),2.93(d,J=9.0Hz,2H),2.39(d,J=9.2Hz,2H),2.24(s,3H) ,2.23(s,3H),2.22(s,3H),2.11(s,3H),2.08(s,2H),1.67(d,J=9.2Hz,4H),1.56–1.51(m,2H),0.84(t,J=7.0Hz,3H); ESI-MS m / z:641.30[M+H] + .

[0122] According to the synthesis method of Example 12, the target compounds in Examples 13-15, 22, 24, 29, 47-50 and 63-66 were prepared.

[0123] Example 13: 5-(1-(1-(cyclopropylmethyl)piperidin-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-13)

[0124] ESI-MS m / z:681.32[M+H] + .

[0125] Example 14: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(1-(1-ethylpiperidin-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-14)

[0126] ESI-MS m / z:655.35[M+H] + .

[0127] Example 15: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-1-(1-(oxetane-3-yl)piperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-15)

[0128] ESI-MS m / z:683.35[M+H] + .

[0129] Example 16: 5-(1-(1-(2,2-difluoroethyl)piperidin-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-16)

[0130] The compound obtained in step 2 of Example 5 was dissolved in DMF (2 mL), and potassium carbonate (0.044 g, 0.32 mmol) and 2,2-difluoroethyl trifluoromethanesulfonate (0.043 g, 0.20 mmol) were added in sequence. The reaction was carried out at 40°C for 2 h. The reaction was monitored to be complete by TLC. 20 mL of water was added, and the mixture was stirred for 30 min and extracted with EA for 3 times. The organic phases were combined, washed with water for 3 times, washed with saturated sodium chloride for 2 times, dried over anhydrous sodium sulfate, evaporated to dryness, and subjected to column chromatography to obtain a white solid (0.039 g, 0.057 mmol). The yield was 35.6%. 1H NMR (600MHz, DMSO-d6) δ11.48(s,1H),8.19(t,J=4.8Hz,1H),7.40(d,J=7.5Hz,2H),7.31(d,J=3.7Hz,2H),7.27(s,1 H),6.28–6.09(m,1H),5.86(s,1H),4.31(d,J=4.8Hz,2H),4.23–4.18(m,1H),3.84(d,J=10.5Hz,2H),3.39(s,3H),3. 25(t,J=11.2Hz,2H),3.12–3.08(m,2H),3.05(d,J=6.9Hz,2H),3.03–2.99(m,1H),2.83–2.77(m,2H),2.40–2.34(m, 4H),2.24(s,3H),2.22(s,3H),2.11(s,3H),1.67(d,J=11.0Hz,4H),1.56–1.51(m,2H),0.85(t,J=6.9Hz,3H); ESI-MS m / z:691.35[M+H] + .

[0131] According to the synthesis method of Example 16, the target compounds in Examples 17, 18, 23, 25, 26, 28, 51, 52 and 67-70 were prepared.

[0132] Example 17: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-2-oxo-1-(1-(2,2,2-trifluoroethyl)piperidin-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-17)

[0133] ESI-MS m / z:709.35[M+H] + .

[0134] Example 18: 5-(1-(1-(cyanomethyl)piperidin-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-18)

[0135] ESI-MS m / z:666.37[M+H] + .

[0136] Example 19: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-1-(1-(methylsulfonyl)azetidin-3-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-19)

[0137] ESI-MS m / z:677.30[M+H] + .

[0138] Example 20: 5-(1-(1-acetylazetidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-20)

[0139] ESI-MS m / z:641.30[M+H] + .

[0140] Example 21: 5-(1-(1-(cyclopropanecarbonyl)azetidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-21)

[0141] ESI-MS m / z:667.30[M+H] + .

[0142] Example 22: 5-(1-(1-(cyclopropylmethyl)azetidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-22)

[0143] ESI-MS m / z:653.37[M+H] + .

[0144] Example 23: 5-(1-(1-(2,2-difluoroethyl)azetidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-23)

[0145] ESI-MS m / z:663.30[M+H] + .

[0146] Example 24: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-1-(1-methylazetidin-3-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-24)

[0147] ESI-MS m / z:613.34[M+H] + .

[0148] Example 25: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(1-(1-(2-hydroxyethyl)azetidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-25)

[0149] ESI-MS m / z:643.35[M+H] + .

[0150] Example 26: 5-(1-(1-(cyanomethyl)azetidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-26)

[0151] ESI-MS m / z:638.33[M+H] + .

[0152] Example 27: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-1-(3-(methylsulfonyl)-3-azabicyclo[3.1.0]hexane-6-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-27)

[0153] ESI-MS m / z:703.32[M+H] + .

[0154] Example 28: 5-(1-(3-(2,2-difluoroethyl)-3-azabicyclo[3.1.0]hexane-6-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-28)

[0155] ESI-MS m / z:689.35[M+H] + .

[0156] Example 29: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(3-methyl-1-(3-methyl-3-azabicyclo[3.1.0]hexane-6-yl)-2-oxo-2,3-dihydro1H-benzo[d]imidazol-5-yl)benzamide (Compound I-29)

[0157] ESI-MS m / z:639.35[M+H] + .

[0158] Example 30: 3-((1-acetylpiperidin-4-yl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-30)

[0159] Step 1: tert-Butyl 4-(5-bromo-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-1-carboxylate

[0160] According to the same preparation method as steps 4, 5 and 6 in Example 1, 4-aminopiperidine-1-carboxylic acid tert-butyl ester was used to replace 4-aminotetrahydropyran to obtain a gray solid. ESI-MS m / z: 395.10 [M+H] + .

[0161] Step 2: tert-Butyl 4-(5-bromo-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-1-carboxylate

[0162] The compound obtained in step 1 of Example 30 (5 g, 12.62 mmol) was used as the raw material and prepared according to the synthetic method of step 8 of Example 1. The yield was 92.6%. ESI-MS m / z: 410.10 [M+H] + .

[0163] Step 3: 5-Bromo-3-methyl-1-(piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one

[0164] The compound obtained in step 2 of Example 30 (4.80 g, 11.69 mmol) was used as the raw material and prepared according to the synthetic method of step 2 of Example 5. Yield: 88.5%. ESI-MS m / z: 310.05 [M+H] + .

[0165] Step 4: 5-Bromo-3-methyl-1-(1-methylpiperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one

[0166] The compound obtained in step 3 of Example 30 (3.18 g, 10.27 mmol) was prepared according to the synthetic method of Example 12, with a yield of 85.6%. ESI-MS m / z: 324.06 [M+H] + .

[0167] Step 5: tert-butyl 4-(ethyl(3-(methoxycarbonyl)-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)amino)piperidine-1-carboxylate

[0168] According to the same preparation method as steps 1, 2 and 3 in Example 1, tetrahydropyrone was replaced with N-tert-butyloxycarbonyl-4-piperidone to obtain a gray solid. ESI-MS m / z: 395.10 [M+H] + .

[0169] Step 6: tert-butyl 4-(ethyl(3-(methoxycarbonyl)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)phenyl)amino)piperidine-1-carboxylate

[0170] Using the compound obtained in step 5 of Example 30 and the compound obtained in step 4 as raw materials, a yellow solid was obtained according to the same preparation method as step 7 of Example 1. ESI-MS m / z: 620.30 [M+H] + .

[0171] Step 7: 3-((1-(tert-butoxycarbonyl)piperidin-4-yl)(ethyl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzoic acid

[0172] The compound obtained in step 6 of Example 30 was prepared in the same manner as in step 9 of Example 1 to obtain a light yellow solid. ESI-MS m / z: 605.30 [M+H] + .

[0173] Step 8: tert-butyl 4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)phenyl)(ethyl)amino)piperidine-1-carboxylate

[0174] The compound obtained in step 7 of Example 30 was prepared in the same manner as in step 10 of Example 1 to obtain a yellow solid. ESI-MS m / z: 739.20 [M+H] + .

[0175] Step 9: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(piperidin-4-yl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro1H-benzo[d]imidazol-5-yl)benzamide

[0176]

[0177] The compound obtained in step 8 of Example 30 was prepared in the same manner as in step 2 of Example 5 to obtain a light yellow solid. ESI-MS m / z: 639.10 [M+H] + .

[0178] Step 10: Preparation of compound I-30

[0179] The compound obtained in step nine of Example 30 was subjected to column chromatography in the same manner as in step three of Example 5 to obtain a white solid with a yield of 32.5%. 1 H NMR (600MHz, DMSO-d6) δ11.47(s,1H),8.18(t,J=4.8Hz,1H),7.40(d,J=3.6Hz,2H),7.33(d,J=8.2Hz,1H),7.30(d,J=8.1Hz,1H),7.27(s ,1H),5.86(s,1H),4.30(d,J=4.7Hz,2H),4.21–4.17(m,1H),3.76(d,J=12.9Hz,1H),3.39(s,3H),3.11–3.07(m,2H),3.05–3.02(m,1H), 3.02–2.95(m,2H),2.92(d,J=10.4Hz,2H),2.54(d,J=11.8Hz,1H),2.41–2.36(m,2H),2.24(s,6H),2.22(s,3H),2.11(s,3H),2.07(d,J= 10.5Hz,2H),1.97(s,3H),1.77–1.72(m,2H),1.65(d,J=10.6Hz,2H),1.51–1.47(m,1H),1.37–1.33(m,1H),0.84(t,J=7.0Hz,3H); ESI-MS m / z:682.40[M+H] + .

[0180] According to the synthetic method of step ten in Example 30, the target compounds in Examples 31, 32 and 41 were prepared.

[0181] Example 31: 3-((1-(cyclopropanecarbonyl)piperidin-4-yl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-31)

[0182] ESI-MS m / z:708.42[M+H] + .

[0183] Example 32: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(1-propionylpiperidin-4-yl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro1H-benzo[d]imidazol-5-yl)benzamide (Compound I-32)

[0184] ESI-MS m / z:696.42[M+H] + .

[0185] Example 33: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(1-methylpiperidin-4-yl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-33)

[0186] The compound of step nine in Example 30 (0.1 g, 0.16 mmol) was used as the raw material and prepared according to the synthesis method of Example 12. Yield: 65.2%. ESI-MS m / z: 653.41 [M+H] + .

[0187] According to the synthetic method of Example 33, Examples 34 and 35 were prepared.

[0188] Example 34: 3-((1-(Cyclopropylmethyl)piperidin-4-yl)(ethyl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)benzamide (Compound I-34)

[0189] ESI-MS m / z:694.44[M+H] + .

[0190] Example 35: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(1-(oxetane-3-yl)piperidin-4-yl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-35)

[0191] ESI-MS m / z:696.42[M+H] + .

[0192] Example 36: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(N-ethylcyclopropanecarboxamido)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-36)

[0193] Step 1: Methyl 5-bromo-3-(ethylamino)-2-methylbenzoate

[0194] Dissolve 3-amino-5-bromo-2-methylbenzoic acid methyl ester (3g, 12.29mmol) in methanol (30mL), add acetaldehyde (0.54g, 12.29mmol), acetic acid (0.74g, 12.29mmol) in turn, and react at room temperature for 30min. Then add NaBH3CN (2.32g, 36.87mmol) in batches, and react at room temperature for 2h. TLC monitors the reaction to be complete. Add water, extract with DCM 3 times, combine the organic phases, wash with saturated sodium chloride 2 times, dry with anhydrous sodium sulfate, evaporate to dryness and column chromatography to obtain a yellow solid (1.18g, 4.33mmol), yield: 35.2%. ESI-MS m / z: 272.02[M+H] + .

[0195] Step 2: 5-bromo-3-(N-ethylcyclopropanecarboxamido)-2-methylbenzoic acid methyl ester

[0196] Dissolve 5-bromo-3-(ethylamino)-2-methylbenzoic acid methyl ester (1.18 g, 4.33 mmol) in DCM (15 mL), add NEt3 (1.31 g, 12.99 mmol), slowly add cyclopropylcarbonyl chloride (0.68 g, 6.50 mmol) in DCM at 0°C. React at room temperature for 2 h. TLC monitors the reaction to be complete. Add water, extract with DCM 3 times, combine the organic phases, wash with saturated sodium chloride 2 times, dry with anhydrous sodium sulfate, and evaporate to obtain a yellow solid (1.41 g, 4.14 mmol), yield: 95.6%. ESI-MS m / z: 340.05 [M+H] + .

[0197] Step 3: Preparation of Compound I-36

[0198] Using the compound obtained in step 2 of Example 36 and the compound obtained in step 4 of Example 30 as raw materials, refer to the synthesis method of Example 1 and obtain a white solid through column chromatography. 1H NMR (600MHz, DMSO-d6) δ11.50(s,1H),8.38(t,J=4.9Hz,1H),7.61(d,J=10.2Hz,2H),7.54(s,1H),7.42–7.39(m, 1H),7.36(d,J=8.2Hz,1H),5.87(s,1H),4.35–4.30(m,2H),4.21(t,J=12.1Hz,1H),3.39(s,3H),2.94(d,J=10.3H z,2H),2.79(s,2H),2.42–2.38(m,2H),2.26(s,3H),2.23(s,3H),2.19(s,3H),2.11(s,3H),1.67(d,J=10.5Hz,2 ESI-MS m / z:625.34[M+H] + .

[0199] Example 37: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-37)

[0200] Step 1: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(4-(methylamino)cyclohexyl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide

[0201]

[0202] According to the synthetic method of step 1 to step 9 of Example 30, a light yellow solid was prepared. ESI-MS m / z: 668.12 [M+H] + .

[0203] Step 2: Preparation of compound I-37

[0204] The compound obtained in step 1 of Example 37 was purified by column chromatography according to the synthesis method of Example 12 to obtain a white solid with a yield of 65.4%. 1H NMR (600MHz, DMSO-d6) δ11.46(s,1H),8.18(t,J=5.0Hz,1H),7.38–7.35(m,2H),7.32(d,J=8.1Hz,1H),7.28–7.26(m,1H),7.23 (d,J=3.3Hz,1H),5.86(s,1H),4.30(d,J=4.8Hz,2H),4.19–4.15(m,1H),3.38(s,3H),3.16–3.06(m,2H),3.05–3.01(m,1H),2. 90(d,J=10.9Hz,2H),2.69–2.64(m,1H),2.39–2.36(m,2H),2.23(s,3H),2.21(s,6H),2.17(s,3H),2.15(s,3H),2.11(s,3H),2 .05(t,2H),1.85–1.78(m,4H),1.65(d,J=10.7Hz,2H),1.40–1.34(m,2H),1.15(q,J=11.6Hz,2H),0.84(t,J=6.4Hz,3H); ESI-MS m / z:682.44[M+H] + .

[0205] According to the synthetic method of Example 37, Examples 38-40 and 42 were prepared.

[0206] Example 38: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(4-(ethyl(methyl)amino)cyclohexyl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-38)

[0207] ESI-MS m / z:696.45[M+H] + .

[0208] Example 39: 3-((4-((cyclopropylmethyl)(methyl)amino)cyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-39)

[0209] ESI-MS m / z:722.47[M+H] + .

[0210] Example 40: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(4-(methyl(oxetan-3-yl)amino)cyclohexyl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-40)

[0211] ESI-MS m / z:724.45[M+H] + .

[0212] Example 41: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(4-(N-methylcyclopropanecarboxamido)cyclohexyl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-41)

[0213] ESI-MS m / z:736.45[M+H] + .

[0214] Example 42: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(4-((2-methoxyethyl)(methyl)amino)cyclohexyl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-42)

[0215] 1H NMR (600MHz, DMSO-d6) δ11.47(s,1H),8.17(t,J=4.8Hz,1H),7.39–7.35(m,2H),7.32(d,J=8.2Hz,1H),7.28(d,J=7.4Hz,1H),7.25–7.22(m,1H),5.8 6(s,1H),4.30(d,J=4.7Hz,2H),4.20–4.15(m,1H),3.38(s,3H),3.33(t,J =6.0Hz,2H),3.20(s,3H),3.12–3.08(m,2H),2.91(d,J=10.6Hz,2H),2.66 (t,J=11.3Hz,1H),2.59–2.54(m,1H),2.41–2.34(m,2H),2.23(s,3H),2.2 2(s,6H),2.21(s,3H),2.15(s,2H),2.11(s,3H),2.06(t,J=11.6Hz,2H),1 .83(d,J=11.1Hz,2H),1.71(d,J=10.7Hz,2H),1.65(d,J=10.8Hz,2H),1.3 8(q,J=11.0Hz,2H),1.19(q,J=11.3Hz,2H),0.84(t,J=6.6Hz,3H); ESI-MS m / z:726.46[M+H] + .

[0216] Example 43: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-2-oxo-3-(tetrahydro-2H-pyran-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-43)

[0217] ESI-MS m / z:628.34[M+H] + .

[0218] Example 44: 5-(3-(1-acetylpiperidin-4-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-44)

[0219] ESI-MS m / z:669.37[M+H] + .

[0220] Example 45: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-3-(1-(methylsulfonyl)piperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-45)

[0221] ESI-MS m / z:705.34[M+H] + .

[0222] Example 46: 5-(3-(1-(cyclopropanecarbonyl)piperidin-4-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-46)

[0223] ESI-MS m / z:695.38[M+H] + .

[0224] Example 47: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-3-(1-(oxetane-3-yl)piperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-47)

[0225] ESI-MS m / z:683.38[M+H] + .

[0226] Example 48: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-3-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-48)

[0227] ESI-MS m / z:641.37[M+H] + .

[0228] Example 49: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(3-(1-ethylpiperidin-4-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-49)

[0229] ESI-MS m / z:655.39[M+H] + .

[0230] Example 50: 5-(3-(1-(cyclopropylmethyl)piperidin-4-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-50)

[0231] ESI-MS m / z:681.40[M+H] + .

[0232] Example 51: 5-(3-(1-(2,2-difluoroethyl)piperidin-4-yl)-1-methyl-2-oxo-2,3-dihydro1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-51)

[0233] ESI-MS m / z:691.37[M+H] + .

[0234] Example 52: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-2-oxo-3-(1-(2,2,2-trifluoroethyl)piperidin-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-52)

[0235] ESI-MS m / z:709.36[M+H] + .

[0236] Example 53: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(3-(1-(ethylsulfonyl)piperidin-4-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-53)

[0237] ESI-MS m / z:719.35[M+H] + .

[0238] Example 54: 5-(3-(1-(cyclopropylsulfonyl)piperidin-4-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-54)

[0239] ESI-MS m / z:731.35[M+H] + .

[0240] Example 55: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-2-oxo-3-(1-propionylpiperidin-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-55)

[0241] ESI-MS m / z:683.38[M+H] + .

[0242] Example 56: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(3-(1-isobutyrylpiperidin-4-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-56)

[0243] ESI-MS m / z:697.40[M+H] + .

[0244] Example 57: 5-(3-(1-acetylazetidin-3-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-57)

[0245] ESI-MS m / z:641.34[M+H] + .

[0246] Example 58: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-3-(1-(methylsulfonyl)azetidin-3-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-58)

[0247] ESI-MS m / z:677.30[M+H] + .

[0248] Example 59: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-2-oxo-3-(1-propionylazetidin-3-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-59)

[0249] ESI-MS m / z:655.35[M+H] + .

[0250] Example 60: 5-(3-(1-(cyclopropylsulfonyl)azetidin-3-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-60)

[0251] ESI-MS m / z:703.32[M+H] + .

[0252] Example 61: 5-(3-(1-(cyclopropanecarbonyl)azetidin-3-yl)-1-methyl-2-oxo-2,3-dihydro1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-61)

[0253] ESI-MS m / z:667.30[M+H] + .

[0254] Example 62: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(3-(1-isobutyrylazetidin-3-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-62)

[0255] ESI-MS m / z:669.37[M+H] + .

[0256] Example 63: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-3-(1-methylazetidin-3-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-63)

[0257] ESI-MS m / z:613.34[M+H] + .

[0258] Example 64: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(3-(1-ethylazetidin-3-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-64)

[0259] ESI-MS m / z:627.36[M+H] + .

[0260] Example 65: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-3-(1-(oxetane-3-yl)azetidin-3-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-65)

[0261] ESI-MS m / z:655.35[M+H] + .

[0262] Example 66: 5-(3-(1-(cyclopropylmethyl)azetidin-3-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-66)

[0263] ESI-MS m / z:653.37[M+H] + .

[0264] Example 67: 5-(3-(1-(2,2-difluoroethyl)azetidin-3-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-67)

[0265] ESI-MS m / z:663.34[M+H] + .

[0266] Example 68: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(1-methyl-2-oxo-3-(1-(2,2,2-trifluoroethyl)azetidin-3-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-68)

[0267] ESI-MS m / z:681.33[M+H] + .

[0268] Example 69: 5-(3-(1-(cyanomethyl)azetidin-3-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-69)

[0269] ESI-MS m / z:638.34[M+H] + .

[0270] Example 70: 5-(3-(1-(cyanomethyl)piperidin-4-yl)-1-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (Compound I-70)

[0271] ESI-MS m / z:666.37[M+H] + .

[0272] Example 71: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-5-(3-ethyl-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-2-methylbenzamide (Compound I-71)

[0273] ESI-MS m / z:642.36[M+H] + .

[0274] Example 72: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-2-methyl-5-(2-oxo-1-(tetrahydro-2H-pyran-4-yl)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-72)

[0275] ESI-MS m / z:614.33[M+H] + .

[0276] Example 73: 3-(Ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro1H-benzo[d]imidazol-5-yl)benzamide (Compound I-73)

[0277] According to the synthesis method of Example 1 and Example 36, a white solid was prepared with a yield of 32.2%. ESI-MS m / z: 657.35 [M+H] + .

[0278] Example 74: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(1-methylpyrrolidin-3-yl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro1H-benzo[d]imidazol-5-yl)benzamide (Compound I-74)

[0279] According to the synthesis method of Example 33, a white solid was prepared with a yield of 22.5%. ESI-MS m / z: 640.0 [M+H] + .

[0280] Example 75: N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(1-(oxetane-3-yl)pyrrolidin-3-yl)amino)-2-methyl-5-(3-methyl-1-(1-methylpiperidin-4-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)benzamide (Compound I-75)

[0281] According to the synthesis method of Example 33, a white solid was prepared with a yield of 15.8%. ESI-MS m / z: 682.0 [M+H] + .

[0282] Biological Activity Test:

[0283] Experiment 1: Determination of the enzyme activity of the compounds of the present invention on EZH2 WT, Y641F and Y641N mutants

[0284] According to the instructions, prepare 1x assay buffer (modified Tris buffer) and use this assay buffer to dilute the enzyme and substrate. Use an appropriate concentration gradient to dilute the compound, and then transfer 100nL of the compound dilution to each well in a 384-well plate. Then transfer 5μL of the enzyme solution to the assay plate, and for the low control, transfer 5μL of 1x assay buffer. Incubate at room temperature for 15min. After incubation, add 5μL of substrate solution to each well to start the reaction and incubate at room temperature for 45min. Then dilute the receptor and donor solutions with 1x assay buffer and add 15μL, incubate at room temperature under weak light for 60min. Read the endpoint on EnSpire using Alpha mode. (This experiment was completed by Shanghai Ruizhi Chemical Research Co., Ltd., and the method is AlphaLISA). The inhibitory activity results of the compounds against EZH2 WT are shown in Table 1. The inhibitory activity results against the EZH2 Y641 mutant are shown in Table 2.

[0285] Table 1 Inhibitory activity of compounds against EZH2 WT

[0286]

[0287] The test results showed that the example compounds had significant inhibition on EZH2 WT at the biochemical level.

[0288] Table 2 Inhibitory activity of some compounds against Y641F and Y641N mutants

[0289] Compound No. Y641F Y641N 12 0.028 0.05 37 0.034 0.083 42 0.14 0.20 EPZ-6438 0.37 0.85 GSK-126 0.33 0.52

[0290] The results showed that the preferred compounds 12, 37 and 42 had strong inhibitory effects on EZH2 Y641F and EZH2 Y641N.

[0291] Experiment 2: Analysis of the effects of the compounds of the present invention on proliferation of human diffuse large B-cell lymphoproliferative cells (Karpas 422 cells)

[0292] The experiment performed CCK-8 analysis on Karpas-422 (Cellcook) cells to evaluate the antiproliferative activity of the target compounds. Karpas 422 cells (about 3000 cells / well) were seeded into RPMI 1640 (Gibco) 96-well plates containing 20% ​​FBS (Cellcook) and 1% penicillin-streptomycin (Gibco). The cells were then treated with compounds with a concentration gradient. First, 50 μL of the test compound was added to the cell plate of each well, and the cell culture was cultured at 37°C and 5% CO2 for 72 hours. After the incubation, the compound was diluted again with growth medium and 50 μL was added to the cell plate of each well. The plate was then cultured for another 96 hours at 37°C and 5% CO2. Then, 50 μL of CCK-8 diluent (including 20 μL of CCK-8 and 30 μL of culture medium) was added to each well and cultured for another 4 hours. Then, the plate was shaken for 5 min and read at 450 nm using a microplate reader (Multiskan FC). All experiments were repeated at least 3 times. Dose-response curves were obtained using Prism 5.0 (GraphPad software). The results of the antiproliferative activity of some examples on Karpas-422 cells are shown in Table 3.

[0293] Table 3 Inhibitory effect of compounds on Karpas-422 proliferation

[0294]

[0295] The results of cell tests showed that the compounds in the examples all had certain antiproliferative activity against Karpas-422. 50 =35.01 nM), some compounds showed stronger antiproliferative activity.

Claims

1. A benzamide compound containing an imidazolidinone structure as shown in general formula I or a pharmaceutically acceptable salt thereof, in R 1 is (C1-C3)alkyl or (C1-C3)alkoxy; R 2 , R 3 Each is independently selected from hydrogen, (C1-C4) alkyl, (C3-C8) cycloalkyl, (C3-C6) cycloalkylacyl, 4-7 membered heterocycloalkyl containing 1-2 atoms selected from N, O and S, wherein the (C3-C8) cycloalkyl may be optionally replaced by 1-3 R 6 When the 4-7 membered heterocycloalkyl group containing 1-2 atoms selected from N, O and S is a heterocycloalkyl group containing sulfur atoms, the sulfur atoms therein may be oxidized to form sulfones; when the 4-7 membered heterocycloalkyl group containing 1-2 atoms selected from N, O and S is a heterocycloalkyl group containing nitrogen atoms, the nitrogen atoms therein may be optionally replaced by 1-2 R 7 replace; R 4 , R 5 Each is independently selected from hydrogen, (C1-C4) alkyl, (C1-C4) alkoxy, (C3-C8) cycloalkyl, 4-7 membered heterocycloalkyl containing 1-2 atoms selected from N, O and S, and a six- to eight-membered bicyclic ring containing 1-2 nitrogen atoms, wherein The (C1-C4) alkyl, (C1-C4) alkoxy, (C3-C8) cycloalkyl may be optionally substituted by 1-3 R 6 When the 4-7 membered heterocycloalkyl containing 1-2 atoms selected from N, O and S is a heterocycloalkyl containing sulfur atoms, the sulfur atoms therein may be oxidized to form sulfones; when the 4-7 membered heterocycloalkyl containing 1-2 atoms selected from N, O and S and the six to eight membered bicyclic ring containing 1-2 nitrogen atoms are heterocycloalkyl containing nitrogen atoms, the nitrogen atoms therein may be optionally replaced by 1-2 R 7 replace; R 6 is selected from hydrogen, halogen, hydroxy, cyano, (C1-C3) alkyl, (C1-C3) alkoxy, (C3-C6) cycloalkyl, optionally substituted by 1-2 R 7 Substituted amino groups; R 7 Selected from hydrogen, halogen, hydroxy, (C1-C4) alkyl, (C1-C4) alkoxy, hydroxy (C1-C4) alkyl, (C4-C6) cycloalkyl, (C1-C4) acyl, (C3-C6) cycloalkyl acyl, (C1-C4) sulfonyl, cyclopropylsulfonyl, (C1-C4) alkyl substituted by 1-3 cyano or fluorine atoms.

2. The benzamide compound containing an imidazolidinone structure or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1 is methyl or methoxy; Selected from:

3. The benzamide compound containing an imidazolidinone structure or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: Selected from:

4. The benzamide compound containing an imidazolidinone structure or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound is selected from compounds I-1 to I-75:

5. The method for preparing the benzamide compound containing an imidazolidinone structure or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, characterized in that: The compound preparation route is selected from route 1 to route 7; Route 1: (a) 4-bromo-1-fluoro-2-nitrobenzene is used as a raw material and reacts with aliphatic amines to obtain intermediate 2 through nucleophilic substitution reaction; (b) using intermediate 2 as a raw material, obtaining intermediate 3 by nitro reduction reaction; (c) Intermediate 3 was used as the starting material to obtain intermediate 4 through a CDI-mediated cyclization reaction; (d) intermediate 5 is used as a raw material, condensed with tetrahydropyrone and reacted with sodium triacetoxyborohydride to obtain intermediate 6; (e) intermediate 6 is used as a raw material, condensed with acetaldehyde and reacted with sodium triacetoxyborohydride to obtain intermediate 7; (f) intermediate 7 is used as a raw material, and intermediate 8 is prepared by Miyaura borate esterification reaction with diboric acid pinacol ester; (g) intermediate 8 is used as a raw material, and intermediate 4 is reacted with intermediate 9 through Suzuki-Miyaura coupling reaction; (h) intermediate 9 is used as a raw material, and reacts with iodomethane or iodoethane to obtain intermediate 10 through nucleophilic substitution reaction; (i) Using intermediate 10 as a raw material, intermediate 11 is obtained by ester hydrolysis under the action of NaOH; (j) using intermediate 11 as a raw material, and 3-(aminomethyl)-4,6-dimethyl-1H-pyridin-2-one hydrochloride or 3-(aminomethyl)-4-methoxy-6-methylpyridin-2(1H)-one hydrochloride, in the presence of a condensing agent, to undergo an amide condensation reaction to obtain the target compound of formula I; Route 2: Using (g), (h), (i) and (j) in route 1, intermediate 12 is reacted in four steps (a), (b), (c) and (d) to obtain the target compound of formula I; Route 3: Wherein, Ring A is an aliphatic heterocycle containing a nitrogen atom, as defined in Formula I; Using the reaction of route 1, intermediate 14 undergoes four steps (a), (b), (c) and (d) to obtain intermediate 18; (e) Using intermediate 18 as a starting material, removing the Boc protecting group under acidic conditions to obtain intermediate 19; (f) intermediate 19 is used as a raw material, reacted with an acyl chloride or sulfonyl chloride through N-acylation or N-sulfonylation reaction, or condensed with a fatty aldehyde or ketone and reacted with sodium triacetoxyborohydride to obtain the target compound of formula I; Route 4: Using the reaction of route 4, the intermediate 20 is reacted through six steps (a), (b), (c), (d), (e) and (f) to obtain the target compound of formula I; Route 5: (a) Intermediate 14 is used as a raw material, and reacts with iodomethane or iodoethane to obtain intermediate 26 through nucleophilic substitution reaction; (b) Using intermediate 26 as the starting material, the Boc protecting group was removed under acidic conditions to obtain intermediate 27; (c) intermediate 27 is used as a raw material, condensed with formaldehyde solution and reacted with sodium triacetoxyborohydride to obtain intermediate 28; Using the reaction in route 3, intermediate 28 is reacted in five steps (d), (e), (f), (g) and (h) to obtain the target compound of formula I; Route 6: Wherein, Ring B is a cycloalkane, as defined in Formula I; Using the reaction in route 5, intermediate 34 is reacted in five steps (a), (b), (c), (d) and (e) to obtain the target compound of formula I; Route 7: (a) Intermediate 5 is used as a raw material, condensed with acetaldehyde and reacted with sodium cyanoborohydride to obtain intermediate 39; (b) intermediate 39 is used as a raw material, and reacted with cycloalkylcarbonyl chloride to obtain intermediate 7 through N-acylation reaction; Using the reaction in route 6, intermediate 7 is reacted in four steps (c), (d), (e) and (f) to obtain the target compound of formula I.

6. A pharmaceutical composition, characterized in that A pharmaceutical composition comprising as an active ingredient the benzamide compound containing an imidazolidinone structure according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, a carrier or an excipient; the pharmaceutical composition is in the form of an injection, a tablet or a capsule.

7. Use of the benzamide compound containing an imidazolidinone structure or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 in the preparation of a medicament for preventing and / or treating diseases associated with abnormal expression of EZH2.

8. Use of the pharmaceutical composition according to claim 6 in the preparation of a medicament for preventing and / or treating diseases associated with abnormal expression of EZH2.

9. The use according to claim 7 or 8, characterized in that: The disease associated with abnormal expression of EZH2 is cancer or autoimmune disease.

10. The use according to claim 9, characterized in that: The cancer is lymphoma, non-small cell lung cancer, small cell lung cancer, head and neck cell carcinoma, bladder cancer, gastric cancer, colon cancer, large intestine cancer, kidney cancer, bile duct cancer, esophageal squamous cell carcinoma, ovarian cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, brain cancer, melanoma, multiple myeloma, glioblastoma, leukemia or cervical cancer; the autoimmune disease is inflammatory bowel disease, autoimmune encephalomyelitis or multiple sclerosis.

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