JQ1 combined pyrimidinopyrrole compound as well as preparation method and application of JQ1 combined pyrimidinopyrrole compound

Abstract

The invention discloses a JQ1 combined pyrimidinopyrrole compound and a preparation method and application thereof.The structural formula of the compound is shown in the formula I. The preparation method of the compound comprises the steps that 1, a compound 1 and a linker protected by t-butyloxycarboryl are subjected to an amidation reaction under the alkaline condition, and an intermediate 2 is obtained; (2) carrying out Buchwald-Harwig coupling reaction on the intermediate 2 and a compound 3 to obtain an intermediate 4; (3) hydrolyzing the intermediate 4 to obtain an intermediate 5; and (4) carrying out amidation reaction on the intermediate 5 and a compound 6 under an alkaline condition to obtain a target product. The pyrrolopyrimidine parent and (+)-JQ-1 are connected in a flexible or rigid ligand mode to obtain a compound with a brand new structure, the activity of the compound on two tumor cell lines of Hela and MDA-MB-231 is superior to that of the (+)-JQ-1, BRD4 protein can be obviously degraded, and the compound has high application value. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a compound, and particularly to a compound of JQ1 binding pyrimidine and pyrrole, and its preparation method and application. Background Art

[0002] (+)-JQ-1 is a traditional small molecule compound that can affect various cancer-related biological processes, including cell proliferation, survival, migration, and differentiation, by inhibiting BRD4 in the BET (Bromodomain and Extra-Terminal domain) family of proteins. BRD4 is a multifunctional protein involved in regulating multiple aspects such as gene transcription, chromatin structure, and DNA damage repair. (+)-JQ-1 can bind to the acetyl-lysine binding pocket of BRD4, preventing it from binding to histone acetylated chromatin, and thus inhibiting BRD4-mediated gene transcription. Since BRD4 is overexpressed in many tumors and is closely related to the occurrence and development of tumors, (+)-JQ-1, as a BRD4 inhibitor, shows potential efficacy in cancer treatment. However, (+)-JQ-1 has not yet been approved as a clinical drug, and its safety and effectiveness need to be addressed urgently.

[0003]

[0004] In recent years, bifunctional small molecules designed and synthesized by the Proteolysis Targeting Chimeras (PROTACs) technology can bring BRD4 protein and intracellular E3 closer, leading to the degradation of the target protein. Compared with traditional inhibitors, such bifunctional small molecules can more effectively inhibit the growth of tumor cells and promote their apoptosis. Developing bifunctional small molecules that can target the ubiquitination and degradation of BRD4 protein is of great significance in studying the function of BRD4 protein and the development of new anti-tumor drugs. Summary of the Invention

[0005] Object of the Invention: The first object of the present invention is to provide a compound of JQ1 binding pyrimidine and pyrrole that can degrade BRD4 protein and exert anti-tumor activity based on the (+)-JQ-1 compound; the second object of the present invention is to provide a preparation method of the compound of JQ1 binding pyrimidine and pyrrole; the third object of the present invention is to provide the application of the compound of JQ1 binding pyrimidine and pyrrole.

[0006] Technical Solution: The compound of JQ1 binding pyrimidine and pyrrole of the present invention has the chemical structural formula as follows:

[0007]

[0008] Among them, Linker is Among them, R1 is H or a three-membered ring; R2 is a 4- to 6-membered nitrogen-containing heterocycle and the nitrogen atom is connected to the carbonyl group, n1 is 0 or 1; a, b, c are selected from 4- to 6-membered nitrogen-containing heterocycles and the nitrogen atom is connected to the carbonyl group; n2 is 1 to 10.

[0009] Preferably, the n2 is 2 to 6.

[0010] Preferably, the is

[0011] Preferably, the is

[0012] Preferably, the is

[0013] Preferably, the structural formula of the compound in which JQ1 binds to pyrimidine and pyrrole is:

[0014]

[0015]

[0016] The preparation method of the compound in which JQ1 binds to pyrimidine and pyrrole according to the present invention includes the following steps:

[0017] (1) 2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid (Compound 1) and Boc-protected linker are subjected to an amidation reaction under alkaline conditions to obtain Intermediate 2;

[0018] (2) Intermediate 2 and 3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline (Compound 3) undergo a Buchwald-Hartwig coupling reaction to obtain Intermediate 4;

[0019] (3) Intermediate 4 is hydrolyzed to obtain Intermediate 5;

[0020] (4) Intermediate 5 and 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid (Compound 6) are subjected to an amidation reaction under alkaline conditions to obtain the target product; The synthesis route is as follows:

[0021]

[0022] Preferably, in step (1), the temperature of the amidation reaction is 0 to 30 °C, and more preferably 10 to 25 °C.

[0023] Preferably, in step (1), the condensing agent for the amidation reaction is 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), 1-butylphosphonic anhydride (T4P), O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU), tetramethylchlorouronium hexafluorophosphate (TCFH), carbonyldiimidazole (CDI), or (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholinium hexafluorophosphate (COMU); preferably TCFH and T4P; the reaction solvent is dimethyl sulfoxide, N,N-dimethylformamide, dichloromethane, acetonitrile, absolute ethanol, preferably acetonitrile and dichloromethane; the reaction base is potassium carbonate, cesium carbonate, triethylamine, N,N-diisopropylethylamine (DIPEA), N-methylimidazole (NMI), preferably N,N-diisopropylethylamine (DIPEA) and N-methylimidazole (NMI).

[0024] Preferably, in step (2), the reaction solvent is dimethyl sulfoxide, N,N-dimethylformamide, 1,4-dioxane, preferably 1,4-dioxane; the reaction catalyst is palladium acetate, palladium chloride, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, preferably palladium acetate; the catalyst ligand is 1,1'-binaphthalene-2,2'-bis(diphenylphosphine) (BINAP), 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthrene (Xanphos), preferably Xanphos; the reaction temperature is 100-150 °C, preferably 100-110 °C; the reaction inert condition is nitrogen or argon protection, preferably nitrogen protection.

[0025] Preferably, in step (3), the reaction acidic reagent is ethyl acetate hydrochloride solution, dichloromethane trifluoroacetate solution, or methanol oxalyl chloride solution, preferably ethyl acetate hydrochloride solution.

[0026] Preferably, in step (4), the condensing agent for the amidation reaction is HATU, HBTU, T4P, TBTU, TCFH, CDI, COMU, preferably TCFH and T4P; the reaction solvent organic solvent is dimethyl sulfoxide, N,N-dimethylformamide, dichloromethane, acetonitrile, absolute ethanol, preferably acetonitrile and dichloromethane; the reaction base is potassium carbonate, cesium carbonate, triethylamine, N,N-diisopropylethylamine (DIPEA), N-methylimidazole (NMI), preferably N,N-diisopropylethylamine (DIPEA) and N-methylimidazole (NMI); the reaction temperature is 0 to -30 °C, preferably 10 to 25 °C.

[0027] Tautomers, optical isomers, deuterated compounds, nitrogen oxides, solvates, pharmaceutically acceptable salts or prodrugs of the JQ1-binding pyrimido[1,2-a]pyrrole compounds of the present invention.

[0028] The salts of the compounds in the present invention preferably include pharmaceutically acceptable salts of the compounds, and the salts can be prepared by any suitable method provided in the literature. For example, inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid are used; or organic acids such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid and salicylic acid are used; pyranose acids such as glucuronic acid and galacturonic acid; α-hydroxy acids such as citric acid and tartaric acid; amino acids such as aspartic acid and glutamic acid; aromatic acids such as benzoic acid and cinnamic acid; sulfonic acids such as p-toluenesulfonic acid and ethanesulfonic acid.

[0029] In the present invention, a pharmaceutically acceptable carrier is relatively non-toxic and harmless to a patient at a concentration consistent with the effective activity of the active ingredient, so that any side effects caused by the carrier do not destroy the beneficial effects of the active ingredient. The pharmaceutically effective amount of the compound or its pharmaceutically acceptable salt is preferably an amount that produces a result or has an impact on the specific condition being treated. The compounds of the present invention can be administered together with pharmaceutically acceptable carriers known in the art in any effective conventional dosage unit form including immediate release, sustained release and timed release formulations in the following ways: oral, parenteral, topical, nasal, ocular, sublingual, rectal, vaginal administration, etc.

[0030] Use of the JQ1-binding pyrimido[1,2-a]pyrrole compounds of the present invention in the preparation of anti-tumor drugs. The compounds of the present invention can effectively degrade the target protein BRD4 in tumor cells.

[0031] The tumors are cervical cancer and breast cancer.

[0032] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages: (1) The present invention connects (+)-JQ-1 and pyrimido[1,2-a]pyrrole compounds through a series of rigid or flexible hydrazine-based Linker structures, targets and degrades BRD4 protein, has good anti-tumor activity, and shows better anti-proliferation and apoptosis-inducing abilities compared with (+)-JQ-1; (2) The IC 50 of the compound against MDA-MB-231 cells can reach 0.22 ± 0.02 μM, and the IC 50 against Hela cells can reach 0.20 ± 0.02 μM. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is a synthetic route diagram of the present invention;

[0034] Figure 2Effect diagram of the compound degrading the target protein BRD4 in tumor cells. Detailed implementation manners

[0035] The technical solution of the present invention will be further described below in conjunction with embodiments.

[0036] Example 1

[0037] The JQ1-binding pyrimidine and pyrrole compound of the present invention, with the chemical name of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-1-(7-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)-4,7-diazaspiro[2.5]octan-4-yl)ethanone, has the following structural formula:

[0038]

[0039] Its preparation method includes the following steps:

[0040] (1) Synthesize tert-butyl 7-(2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)-4,7-diazaspiro[2.5]octane-4-carboxylate, with the following structural formula:

[0041]

[0042] Add 800 mg (3.01 mmol) of 2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid, 1011 mg (3.61 mmol) of TCFH, 839 μL (10.54 mmol) of NMI, 767 mg (3.61 mmol) of tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate and 10 mL of acetonitrile into the reaction flask in sequence, stir at 25 - 35 °C for 2 hours, concentrate the reaction solution, and purify by column chromatography (petroleum ether:ethyl acetate = 7:1) to obtain 687 mg of the target product, with a yield of 49.60%.

[0043] (2) Synthesize tert-butyl 7-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)-4,7-diazaspiro[2.5]octane-4-carboxylate, with the following structural formula:

[0044]

[0045] To the reaction flask, 432 mg (1.79 mmol) of 3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline, 1460 mg (4.48 mmol) of Cs2CO3, 686 mg (1.49 mmol) of the yellow product from the previous step, 432 mg (0.75 mmol) of Xantphos, 84 mg (0.37 mmol) of Pd(OAc)2 and 10 mL of 1,4-dioxane were added successively. The reaction was carried out at 110 °C for 6 hours. After suction filtration and concentration under reduced pressure, the concentrate was purified by column chromatography (DCM:MeOH = 50:1) to obtain 911 mg of the target product with a yield of 91.76%.

[0046] (3) Synthesis of (7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)(4,7-diazaspiro[2.5]octan-7-yl)methanone, the structural formula is as follows:

[0047]

[0048] To the reaction product from the previous step, 10 mL of hydrochloric acid ethyl acetate solution was added, and the reaction was carried out at 25 - 35 °C for 2 hours. After concentration under reduced pressure, 760 mg of the target product was obtained with a yield of 98.22%.

[0049] (4) Synthesis of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-1-(7-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)-4,7-diazaspiro[2.5]octan-4-yl)ethan-1-one

[0050] Add 200 mg (0.35 mmol) of 7-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)-4,7-diazaspiro[2.5]octane-4-carboxylic acid, 108 mg (0.39 mmol) of TCFH, 89 μL (1.12 mmol) of NMI, 129 mg (0.32 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid and 10 mL of acetonitrile into the reaction flask in sequence. Stir at 25 - 35 °C for 2 h, monitor the reaction by TLC until completion (DCM:MeOH = 10:1), and purify by column chromatography (DCM:MeOH = 30:1) to obtain 22 mg of the target product with a yield of 7.21%.

[0051] The target compound is a white solid; 1 H NMR (400 MHz, Methanol-d4) δ 8.73 (s, 1H), 8.21 (d, J = 6.1 Hz, 1H), 8.13–8.05 (m, 2H), 7.42 (q, J = 8.6 Hz, 6H), 7.37–7.30 (m, 1H), 6.65 (s, 1H), 4.79 (d, J = 7.4 Hz, 1H), 4.72 (t, J = 7.0 Hz, 1H), 4.53–4.16 (m, 2H), 3.95 (m, 2H), 3.79 (d, J = 35.0 Hz, 2H), 3.63 (d, J = 3.4 Hz, 2H), 2.68 (d, J = 5.3 Hz, 3H), 2.48 (d, J = 20.1 Hz, 2H), 2.41 (s, 3H), 2.26 (s, 3H), 2.06 (m, 2H), 1.82 (m, 2H), 1.66 (s, 3H), 1.61 (m, 2H), 1.36–1.20 (m, 4H). 1313C NMR (101 MHz, MeOD) δ 174.72, 172.92, 167.04, 165.78, 157.75, 157.31, 154.14, 154.00, 152.75, 145.20, 140.76, 140.21, 138.85, 138.63, 136.93, 134.26, 133.94, 133.77, 133.62, 132.66, 132.44, 131.91, 130.48, 127.22, 124.52, 116.89, 116.82, 115.54, 115.21, 114.87, 111.66, 103.36, 59.86, 56.44, 42.00, 38.30, 33.49, 32.08, 31.39, 28.74, 27.35, 25.73, 24.38, 15.01, 13.89, 13.56, 12.26.

[0052] Example 2

[0053] The JQ1-binding pyrimido-pyrrole compound of the present invention has the chemical name of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-1-(7-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)-2,7-diazaspiro[4.4]nonan-2-yl)ethanone, and the structural formula is as follows:

[0054]

[0055] Its preparation method comprises the following steps:

[0056] (1) On the basis of Example 1, in step (1), 2-tert-butoxycarbonyl-2,7-diazaspiro[4.4]nonane is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the other conditions remain unchanged. Steps (2) and (3) are the same as those in Example 1, and the intermediate (7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)(2,7-diazaspiro[4.4]nonan-2-yl)methanone (M = 578.64) is obtained, and the structural formula is as follows:

[0057]

[0058] (4) Add 200 mg (0.34 mmol) of the intermediate prepared in step (3), 134 mg (1.04 mmol) of DIPEA, and 138 mg (0.35 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid into the reaction flask in sequence. Add 10 mL of dichloromethane as the solvent, add 186 mg (0.52 mmol) of T4P at 0 °C, stir evenly, then transfer to 25 - 30 °C and react for 2 hours. Monitor the reaction by TLC until it is completed (DCM:MeOH = 10:1). Concentrate under reduced pressure and purify by column chromatography (DCM:MeOH = 30:1) to obtain 60 mg of the target product, with a yield of 18.35%.

[0059] The target compound is a yellow powder; 1 HNMR (400 MHz, Methanol-d4) δ 8.71 (dd, J = 13.3, 2.8 Hz, 1H), 8.19 (q, J = 5.3, 3.8 Hz, 1H), 8.12 (s, 1H), 8.08–8.02 (m, 1H), 7.46–7.28 (m, 5H), 7.22 (dt, J = 18.5, 8.3 Hz, 1H), 6.83–6.70 (m, 1H), 4.66 (ddd, J = 9.3, 6.8, 2.9 Hz, 1H), 3.90 (d, J = 14.5 Hz, 1H), 3.86–3.73 (m, 4H), 3.68–3.45 (m, 6H), 2.69–2.60 (m, 3H), 2.44–2.31 (m, 6H), 2.27–2.24 (m, 3H), 2.14 (s, 2H), 2.02 (d, J = 11.3 Hz, 2H), 1.79 (s, 2H), 1.66–1.60 (m, 2H), 1.58–1.51 (m, 2H), 1.26 (dd, J = 13.2, 5.7 Hz, 2H). 1313C NMR (101 MHz, MeOD) δ 171.49, 171.36, 166.50, 164.39, 157.29, 157.18, 156.95, 153.75, 153.54, 152.35, 144.76, 140.33, 139.75, 138.18, 136.48, 134.20, 133.78, 133.49, 133.36, 132.23, 132.12, 131.50, 131.26, 130.11, 129.99, 129.89, 126.80, 124.09, 116.43, 116.37, 115.05, 114.73, 114.37, 111.14, 103.87, 59.28, 55.41, 46.66, 46.27, 35.94, 31.65, 31.52, 25.23, 14.56, 13.48, 13.11, 11.81.

[0060] Example 3

[0061] The JQ1-binding pyrimido[2,3-d]pyrrole compound of the present invention has the chemical name of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-1-(4-((1-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)piperidin-4-yl)methyl)piperazin-1-yl)ethanone, and the structural formula is as follows:

[0062]

[0063] Its preparation method comprises the following steps:

[0064] (1) On the basis of Example 1, in step (1), tert-butyl 4-(piperazin-1-ylmethyl)piperidine-1-carboxylate is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the rest of the conditions remain unchanged. Steps (2) and (3) are the same as those in Example 1, to obtain the intermediate (7-cyclopentyl-2-[(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino]-7H-pyrrolo[2,3-d]pyrimidin-6-yl)(4-(piperazin-1-ylmethyl)piperidin-1-yl)methanone, and the structural formula is as follows:

[0065]

[0066] (4) Add 200 mg (0.31 mmol) of the intermediate prepared in step (3), 138 mg (0.35 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid, 151 mg (1.16 mmol) of DIPEA, and 10 mL of dichloromethane to the reaction flask in sequence. Dropwise add 133 μL (0.44 mmol) of T4P under ice bath conditions. After stirring evenly, transfer the reaction mixture to 25 - 35 °C and react for 2 hours. Monitor the reaction by TLC until it is completed (DCM:MeOH = 10:1). Concentrate under reduced pressure and purify by column chromatography (DCM:MeOH = 30:1) to obtain 25 mg of the target product with a yield of 8.5%.

[0067] The target compound is a white solid; 1 H NMR (400 MHz, Methanol-d4) δ 8.73 (d, J = 2.0 Hz, 1H), 8.21 (t, J = 2.0 Hz, 1H), 8.13–8.04 (m, 2H), 7.42–7.27 (m, 6H), 6.58 (d, J = 4.2 Hz, 1H), 4.73 (dt, J = 10.6, 7.8 Hz, 1H), 4.15–3.97 (m, 1H), 3.91–3.39 (m, 8H), 3.30–3.16 (m, 1H), 3.04–2.86 (m, 1H), 2.68 (s, 3H), 2.60–2.45 (m, 4H), 2.41 (s, 3H), 2.38–2.17 (m, 8H), 2.08 (s, 1H), 2.06–2.00 (m, 2H), 1.94 (d, J = 10.5 Hz, 2H), 1.90–1.75 (m, 4H), 1.64 (d, J = 23.8 Hz, 4H). 13CNMR (101 MHz, MeOD) δ 170.16, 169.22, 164.75, 163.00, 155.78, 155.15, 151.81, 150.68, 143.22, 138.73, 138.18, 138.16, 136.78, 136.54, 134.90, 132.43, 132.10, 131.75, 130.65, 129.93, 128.38, 125.20, 122.49, 114.86, 113.49, 113.14, 112.95, 109.59, 100.24, 63.63, 57.84, 53.97, 53.38, 53.31, 52.93, 52.83, 46.00, 45.36, 41.55, 34.53, 33.17, 31.41, 31.08, 29.99, 23.61, 19.70, 12.95, 11.86, 11.79, 11.52, 10.20.

[0068] Example 4

[0069] The JQ1-binding pyrimido[2,3-d]pyrrole compound of the present invention has the chemical name of N-(7-(2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)-7-azaspiro[3.5]nonan-2-yl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide, and the structural formula is as follows:

[0070]

[0071] Its preparation method comprises the following steps:

[0072] (1) On the basis of Example 1, in step (1), 2-amino-7-tert-butoxycarbonyl-7-azaspiro[3.5]nonane is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the other conditions remain unchanged. Steps (2) and (3) are the same as those in Example 1, and the intermediate 7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-N-(7-azaspiro[3.5]nonan-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide is obtained, and the structural formula is as follows:

[0073]

[0074] (4) Add 348 mg (0.59 mmol) of the intermediate prepared in step (3), 180 mg (0.64 mmol) of TCFH, 149 mg (1.87 mmol) of NMI, and 214 mg (0.53 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid to the reaction flask in sequence, then add 10 mL of dichloromethane, stir at 25 - 30 °C for 2 h, monitor by TLC until the reaction is complete (DCM:MeOH = 20:1), concentrate under reduced pressure, and purify by column chromatography (DCM:MeOH = 30:1) to obtain 96 mg of the target product with a yield of 16.7%.

[0075] The target compound is a white solid; 1 HNMR (400 MHz, Methanol-d4) δ 8.74 (s, 1H), 8.19 (s, 1H), 8.11 (s, 1H), 8.06 (t, J = 1.1 Hz, 1H), 7.45–7.35 (m, 5H), 7.31 (d, J = 1.6 Hz, 1H), 6.91 (d, J = 2.8 Hz, 1H), 5.42–5.29 (m, 1H), 4.75–4.67 (m, 1H), 4.49 (td, J = 8.2, 4.0 Hz, 1H), 3.75–3.47 (m, 6H), 2.68 (s, 3H), 2.46 (dd, J = 20.2, 10.0 Hz, 2H), 2.41 (s, 6H), 2.27 (d, J = 1.1 Hz, 3H), 1.96 (dt, J = 21.4, 10.7 Hz, 4H), 1.83 (m, 4H), 1.67 (s, 6H). 1313C NMR (101 MHz, MeOD) δ 171.13, 171.08, 166.71, 164.44, 157.86, 157.48, 154.60, 154.49, 152.68, 145.13, 140.76, 140.19, 138.84, 138.57, 136.91, 134.97, 134.26, 134.13, 133.94, 133.76, 132.71, 132.63, 131.95, 130.41, 127.22, 124.51, 116.87, 115.60, 115.28, 114.51, 111.71, 105.44, 59.12, 56.08, 45.00, 44.73, 41.99, 41.28, 41.02, 40.67, 37.97, 37.40, 36.65, 34.35, 31.78, 25.74, 14.98, 13.89, 13.55, 12.23.

[0076] Example 5

[0077] The JQ1-binding pyrimido[2,3-d]pyrrole compound of the present invention has the chemical name of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-1-(4-(4-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)piperazin-1-yl)methyl)piperidin-1-yl)ethanone, and its structural formula is as follows

[0078]

[0079] Its preparation method comprises the following steps:

[0080] (1) On the basis of Example 1, in step (1), 1-tert-butoxycarbonyl-4-(piperazin-1-ylmethyl)piperidine is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the rest of the conditions remain unchanged. Steps (2) and (3) are the same as those in Example 1, and the intermediate (7-cyclopentyl-2-[(3-(4-methyl-1H-imidazo[1,1-d]pyrimidin-5-(trifluoromethyl)phenyl)amino]-7H-pyrrolo[2,3-d]pyrimidin-6-yl)(4-(piperidin-4-ylmethyl)piperazin-1-yl)methanone is obtained, and its structural formula is as follows:

[0081]

[0082] (4) Add 140 mg (0.22 mmol) of the intermediate prepared in step (3), 80 mg (0.2 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,10-trimethyl-6,7-dihydro-3H-1l4-thieno[3,2-g][1,2,4]triazolo[4,3-a][1,5]diazocin-6-yl), 10 mL of dichloromethane, 67 mg (0.2 mmol) of TCFH, and 57.4 mg (0.7 mmol) of NMI to the reaction flask in sequence. Stir at 25 - 30 °C for 2 hours and monitor by TLC (DCM:MeOH = 20:1) until the reaction is completed. Concentrate under reduced pressure and purify by column chromatography (DCM:MeOH = 35:1) to obtain 30 mg of the target product with a yield of 14.73%.

[0083] The target compound is a white solid; 1 H NMR (400 MHz, DMSO-d6) δ 9.95 (s, 1H), 8.75 (s, 1H), 8.22 (d, J = 12.4 Hz, 2H), 8.05 (d, J = 4.9 Hz, 1H), 7.38 (t, J = 8.9 Hz, 6H), 6.53 (s, 1H), 4.66 (q, J = 9.0 Hz, 1H), 4.49 (t, J = 6.7 Hz, 1H), 4.27 (d, J = 12.6 Hz, 1H), 4.03 (d, J = 14.0 Hz, 6H), 2.42 (s, 3H), 2.31 (s, 6H), 2.24 (m, 2H), 2.07 (d, J = 12.0 Hz, 6H), 1.88 (s, 2H), 1.78–1.71 (m, 2H), 1.64 (d, J = 8.4 Hz, 2H), 1.53 (s, 3H), 1.48–1.40 (m, 2H), 1.14 (d, J = 6.5 Hz, 4H), 0.94–0.68 (m, 2H). 1313C NMR (101 MHz, DMSO) δ 168.25, 163.29, 162.05, 155.80, 155.39, 152.71, 151.68, 150.20, 143.51, 139.12, 138.45, 137.27, 135.66, 135.42, 132.61, 132.31, 131.15, 130.59, 130.35, 130.12, 128.92, 125.67, 122.96, 114.81, 113.49, 112.77, 101.07, 79.77, 79.65, 79.44, 79.13, 79.05, 63.91, 57.31, 54.67, 48.34, 48.12, 47.93, 47.78, 45.65, 41.83, 35.27, 33.19, 30.47, 29.47, 29.16, 24.02, 14.45, 13.99, 13.12, 11.70.

[0084] Example 6

[0085] The JQ1-binding pyrimidine and pyrrole compound of the present invention, with the chemical name of 2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-1-(3-(4-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)piperazin-1-yl)azetidin-1-yl)ethanone, has the following structural formula:

[0086]

[0087] Its preparation method includes the following steps:

[0088] (1) On the basis of Example 1, in step (1), 1-Boc-3-(piperazin-1-yl)azetidine is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the other conditions remain unchanged. Steps (2) and (3) are the same as in Example 1, to obtain the intermediate (4-(azetidin-3-yl)piperazin-1-yl)(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)methanone, with the following structural formula:

[0089]

[0090] (4) Add 200 mg (0.34 mmol) of the intermediate prepared in step (3), 80 mg (0.2 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,10-trimethyl-6,7-dihydro-3H-1l4-thieno[3,2-g][1,2,4]triazolo[4,3-a][1,5]diazocin-6-yl), 10 mL of dichloromethane, 67 mg (0.2 mmol) of TCFH, and 57.4 mg (0.7 mmol) of NMI into the reaction flask in sequence. Stir at 25 - 30 °C for 2 hours and monitor by TLC (DCM:MeOH = 10:1) until the reaction is complete. Concentrate under reduced pressure and purify by column chromatography (DCM:MeOH = 30:1) to obtain 61 mg of the target product with a yield of 31.23%.

[0091] The target compound is a white solid; 1 H NMR (400 MHz, DMSO-d6) δ 9.95 (s, 1H), 8.75 (s, 1H), 8.22 (d, J = 12.4 Hz, 2H), 8.05 (d, J = 4.9 Hz, 2H), 7.38 (t, J = 8.9 Hz, 5H), 6.53 (s, 1H), 4.66 (q, J = 9.0 Hz, 1H), 4.49 (t, J = 6.7 Hz, 1H), 4.27 (d, J = 12.6 Hz, 1H), 4.13–3.94 (m, 4H), 3.52 (dt, J = 16.3, 8.1 Hz, 4H), 3.12–2.92 (m, 2H), 2.42 (s, 3H), 2.34–2.24 (m, 7H), 2.07 (d, J = 12.0 Hz, 6H), 1.89 (d, J = 9.8 Hz, 2H), 1.79 (m, 2H), 1.68–1.62 (m, 2H), 1.48–1.41 (m, 2H). 13CNMR (101 MHz, DMSO) δ 170.28, 170.20, 163.67, 162.17, 162.14, 155.43, 155.35, 152.73, 151.73, 150.38, 150.36, 143.52, 143.41, 139.13, 138.46, 137.20, 135.73, 135.42, 132.67, 132.20, 131.40, 131.19, 130.60, 130.34, 130.31, 130.00, 128.94, 125.68, 122.97, 114.79, 113.49, 112.76, 112.59, 109.33, 101.24, 79.66, 79.12, 57.33, 54.28, 53.39, 51.60, 33.82, 30.48, 24.05, 19.11, 14.46, 14.02, 13.11, 11.72.

[0092] Example 7

[0093] The JQ1-binding pyrimidine and pyrrole compound of the present invention has the chemical name of 2-((6S,Z)-4-(4-chlorophenyl)-2,3,10-trimethyl-6,7-dihydro-3H-1l4-thieno[3,2-g][1,2,4]triazolo[4,3-a][1,5]diazepin-6-yl)-1-(8-(7-cyclopentyl-2-)((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)-2,8-diazaspiro[4.5]dec-2-yl)acetic acid-1-ketone, and its structural formula is as follows:

[0094]

[0095] Its preparation method includes the following steps:

[0096] (1) On the basis of Example 1, in step (1), 2-tert-butoxycarbonyl-2,8-diazaspiro[4.5]decane is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the remaining conditions remain unchanged. Steps (2) and (3) are the same as those in Example 1, and the intermediate (7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)(2,8-diazaspiro[4.5]dec-8-yl)methanone is obtained, and its structural formula is as follows:

[0097]

[0098] (4) Add 200 mg (0.34 mmol) of the intermediate prepared in step (3), 10 mL of dichloromethane, 67 mg (0.2 mmol) of TCFH, and 57.4 mg (0.7 mmol) of NMI to the reaction flask in sequence, stir at 25 - 30 °C for 2 hours, monitor by TLC (DCM:MeOH = 10:1) until the reaction is completed, concentrate under reduced pressure, and purify by column chromatography (DCM:MeOH = 35:1) to obtain 147 mg of the target product with a yield of 47.82%.

[0099] The target compound is a slightly yellow solid; 1 HNMR (400 MHz, DMSO-d6) δ 10.08 (s, 1H), 8.86 (d, J = 1.9 Hz, 1H), 8.31 (d, J = 2.5 Hz, 1H), 8.19 (s, 2H), 7.47 (q, J = 8.7, 6.6 Hz, 6H), 6.66 (d, J = 2.5 Hz, 1H), 4.79 (td, J = 8.8, 3.6 Hz, 1H), 4.65 (td, J = 6.9, 3.5 Hz, 1H), 3.77–3.66 (m, 8H), 3.56 (d, J = 8.0 Hz, 5H), 3.46–3.25 (m, 4H), 2.75 (t, J = 4.7 Hz, 2H), 2.60 (d, J = 4.1 Hz, 3H), 2.38 (s, 3H), 2.21 (s, 3H), 1.99 (dd, J = 18.7, 7.8 Hz, 2H), 1.78 (d, J = 8.6 Hz, 2H), 1.67 (m, 2H), 1.61 (s, 3H), 1.60–1.51 (m, 2H). 13 C NMR (101 MHz, DMSO) δ 168.86, 168.78, 163.50, 162.06, 162.02, 155.64, 155.35, 152.54, 151.65, 150.28, 143.53, 139.10, 138.44, 137.27, 137.23, 135.68, 135.39, 132.76, 132.65, 131.40, 131.08, 130.55, 130.31, 130.01, 128.86, 125.68, 122.97, 114.78, 113.40, 112.83, 112.55, 109.27, 100.53, 65.23, 57.36, 54.50, 54.30, 44.76, 44.05, 41.70, 36.89, 36.47, 35.61, 34.69, 30.44, 24.05, 14.39, 14.32, 13.97, 13.03, 11.67.

[0100] Example 8

[0101] The JQ1-binding pyrimido[2,3-d]pyrrole compound of the present invention has the chemical name ((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin- -6-yl)-1-(4-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)piperazin-1-yl)ethanone, and its structural formula is as follows:

[0102]

[0103] Its preparation method comprises the following steps:

[0104] (1) On the basis of Example 1, in step (1), N-Boc piperazine is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the remaining conditions remain unchanged. Steps (2) and (3) are the same as those in Example 1, and the intermediate (7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)(piperazin-1-yl)methanone is obtained, and its structural formula is as follows:

[0105]

[0106] (4) 300 mg (0.56 mmol) of the intermediate prepared in step (3), 202 mg (0.5 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin- -6-yl)acetic acid, 187 mg (0.67 mmol) of TCFH, 10 mL of dichloromethane, 155 mg (1.90 mmol) of NMI are added to the reaction flask in sequence, stirred at 25-30 °C for 2 hours, monitored by TLC (DCM:MeOH = 10:1) until the reaction is completed, concentrated under reduced pressure, and purified by column chromatography (DCM:MeOH = 40:1) to obtain 85 mg of the target product, with a yield of 18.40%.

[0107] The target compound is a white solid; 11H NMR (400 MHz, DMSO-d6) δ 10.10 (s, 1H), 8.88 (s, 1H), 8.35 (d, J = 13.3 Hz, 2H), 8.17 (s, 1H), 7.51 (dd, J = 11.8, 9.6 Hz, 4H), 7.45 (d, J = 8.5 Hz, 2H), 6.74 (s, 1H), 4.87–4.75 (m, 1H), 4.61 (d, J = 13.4 Hz, 1H), 3.79 (s, 1H), 3.66–3.52 (m, 8H), 3.12 (dd, J = 7.3, 4.0 Hz, 3H), 2.61 (s, 3H), 2.42 (s, 3H), 2.33 (m, 2H), 2.22 (s, 3H), 2.03–1.94 (m, 2H), 1.79–1.67 (m, 2H), 1.64 (s, 3H), 1.58–1.47 (m, 2H). 13 13C NMR (101 MHz, DMSO) δ 169.04, 163.44, 162.47, 155.67, 155.41, 152.89, 151.74, 150.28, 143.53, 138.22, 138.10, 137.22, 135.68, 135.34, 132.66, 132.12, 131.19, 130.36, 130.09, 128.95, 125.65, 122.94, 118.13, 115.28, 113.79, 112.97, 112.83, 109.68, 101.57, 72.79, 70.24, 60.67, 57.40, 54.61, 53.75, 42.40, 35.31, 31.76, 30.46, 24.04, 18.42, 17.15, 14.47, 13.53, 12.62, 11.73.

[0108] Example 9

[0109] The JQ1-binding pyrimidine and pyrrole compound of the present invention, with the chemical name 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin -6-yl)-1-(6-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)-2,6-diazaspiro[3.3]heptan-2-yl)ethanone has the following structural formula:

[0110]

[0111] The preparation method comprises the following steps:

[0112] (1) Based on Example 1, step (1) uses 2-tert-butyloxycarbonyl-2,6-diazaspiro[3.3]heptane to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the other conditions remain unchanged. Steps (2) and (3) are the same as in Example 1 to obtain the intermediate 7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)(2,6-diazaspiro[3.3]heptane-2-yl)methanone, the structural formula of which is as follows:

[0113]

[0114] (4) 100 mg (0.25 mmol) of the intermediate prepared in step (3), 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepine were added to the reaction flask in sequence. 158 mg (0.28 mmol) of 2-(6-yl)acetic acid, 119 mg (0.74 mmol) of DIPEA, 10 mL of dichloromethane, 119 mg (0.38 mmol) of T4P, react at 25-30 °C for 3 h, monitor the reaction by TLC until completion (DCM:MeOH=10:1), concentrate under reduced pressure, and purify by column chromatography (DCM:MeOH=40:1) to obtain 85 mg of the product with a yield of 36.35%.

[0115] The target compound is a white solid; 1 H NMR (400MHz, DMSO-d6) δ10.12(d,J=6.2Hz,1H),8.85(s,1H),8.29(s,1H),8.14(d,J=19.1Hz ,2H),7.64(s,1H),7.52–7.35(m,4H),7.14(s,1H),6.95–6.85(m,1H),5.24(t,J=9.8Hz,1H), 4.54(d,J=6.5Hz,1H),3.88(d,J=10.4Hz,1H),3.66(s,8H),3.44–3.28(m,3H),2.59(d,J=6.4 Hz,3H),2.39(dd,J=10.2,6.0Hz,3H),2.19(d,J=4.0Hz,3H),1.95(m,2H),1.84–1.15(m,9H). 1313C NMR (101 MHz, DMSO) δ 171.08, 163.56, 163.03, 155.53, 153.46, 152.17, 150.25, 143.37, 139.12, 138.46, 137.11, 135.67, 135.45, 132.72, 131.13, 130.55, 130.27, 130.00, 129.71, 128.90, 128.50, 125.66, 122.95, 121.08, 114.83, 113.72, 112.78, 112.51, 109.56, 104.55, 72.81, 70.46, 70.24, 69.98, 60.67, 56.91, 54.23, 38.08, 33.34, 31.77, 30.24, 24.02, 19.29, 14.48, 14.02, 13.06, 11.75.

[0116] Example 10

[0117] The JQ1-binding pyrimidine and pyrrole compound of the present invention, with the chemical name of synthetic 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin -6-yl)-1-(9-(7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonyl)-3,9-diazaspiro[5.5]undecan-3-yl)ethan-1-one, and the structure is as follows:

[0118]

[0119] Its preparation method comprises the following steps:

[0120] (1) On the basis of Example 1, in step (1), 3-tert-butoxycarbonyl-3,9-diazaspiro[5.5]undecane- is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the other conditions remain unchanged. Steps (2) and (3) are the same as in Example 1, and the intermediate (7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)--(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine--yl)(3,9-diazaspiro[5.5]undecan-3-yl)methanone is obtained. The structural formula is as follows:

[0121]

[0122] (4) Add 300 mg (0.56 mmol) of the intermediate prepared in step (3), 187 mg (0.67 mmol) of TCFH, 10 mL of acetonitrile, 155 mg (1.9 mmol) of NMI, and 202 mg (0.5 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid to the reaction flask in sequence, and react at 25 - 30 °C for 2 hours. Monitor the reaction by TLC until it is completed (DCM:MeOH = 10:1). Concentrate under reduced pressure and purify by column chromatography (DCM:MeOH = 40:1) to obtain 150 mg of the product, with a yield of 30.25%. The target compound is a white solid;

[0123] H NMR (400 MHz, DMSO-d6) δ 10.04 (d, J = 2.0 Hz, 1H), 8.84 (d, J = 3.1 Hz, 1H), 8.30 (q, J = 2.1 Hz, 1H), 8.18–8.09 (m, 2H), 7.53–7.40 (m, 5H), 7.15–6.87 (m, 1H), 6.62 (d, J = 6.6 Hz, 1H), 4.80–4.69 (m, 1H), 4.59 (dd, J = 7.7, 5.8 Hz, 1H), 3.49 (ddd, J = 12.6, 6.3, 3.5 Hz, 3H), 3.40 (ddd, J = 10.4, 6.8, 4.0 Hz, 9H), 2.60 (s, 3H), 2.41 (s, 3H), 2.38–2.27 (m, 2H), 2.18 (s, 3H), 1.98 (s, 2H), 1.80–1.67 (m, 2H), 1.63 (s, 3H), 1.59–1.44 (m, 8H), 1.42–1.28 (m, 2H). 1 13 1313C NMR (101 MHz, DMSO) δ 168.37, 163.30, 161.99, 155.78, 155.33, 152.60, 151.61, 150.20, 143.53, 139.12, 138.49, 137.25, 135.65, 135.45, 132.83, 131.13, 130.61, 130.34, 130.10, 129.25, 128.94, 116.06, 114.82, 114.31, 113.50, 112.85, 109.34, 100.40, 72.81, 70.46, 70.25, 69.99, 63.26, 60.68, 57.31, 54.70, 49.06, 37.44, 35.74, 35.23, 31.77, 31.05, 30.45, 28.56, 24.06, 21.79, 19.30, 14.48, 14.06, 13.15, 11.74.

[0124] Example 11

[0125] The JQ1-binding pyrimido[2,3-d]pyrrole compound of the present invention has the chemical name of N-[2-[2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl]-2-azaspiro[3.3]heptan-6-yl]-7-cyclopentyl-2-[[3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl]amino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide, and the structural formula is as follows:

[0126]

[0127] Its preparation method comprises the following steps:

[0128] (1) On the basis of Example 1, in step (1), 2-tert-butoxycarbonyl-6-amino-2-azaspiro[3.3]heptane is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the remaining conditions remain unchanged. Steps (2) and (3) are the same as those in Example 1, and the intermediate 7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-N-(2-azaspiro[3.3]heptan-6-yl)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide is obtained, and the structural formula is as follows:

[0129]

[0130] (4) Add 300 mg (0.56 mmol) of the intermediate prepared in step (3), 187 mg (0.67 mmol) of TCFH, 10 mL of acetonitrile, 155 mg (1.90 mmol) of NMI, and 202 mg (0.5 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid to the reaction flask in sequence, and react at 25 - 30 °C for 2 hours. Monitor the reaction by TLC until it is completed (DCM:MeOH = 10:1). Concentrate under reduced pressure and purify by column chromatography (eluent DCM:MeOH = 40:1) to obtain 180 mg of the product, with a yield of 37.91%. The target compound is a white solid; the total yield is 45%;

[0131] HNMR (400 MHz, DMSO-d6) δ 10.10 (d, J = 4.9 Hz, 1H), 8.87 (d, J = 8.5 Hz, 1H), 8.29 (s, 1H), 8.11 (s, 1H), 7.96 (s, 1H), 7.45 (dt, J = 19.0, 8.1 Hz, 4H), 7.01 (dd, J = 17.6, 10.1 Hz, 2H), 6.56 (d, J = 7.6 Hz, 1H), 6.49 (t, J = 7.2 Hz, 1H), 5.52–5.35 (m, 1H), 4.58–4.50 (m, 1H), 4.49–4.34 (m, 1H), 4.02–3.55 (m, 5H), 3.20–3.11 (m, 3H), 2.89 (s, 3H), 2.73 (s, 3H), 2.60 (d, J = 3.0 Hz, 2H), 2.40 (d, J = 6.1 Hz, 4H), 2.19 (s, 3H), 1.91 (s, 2H), 1.79–1.69 (m, 2H), 1.54 (dd, J = 55.1, 9.7 Hz, 5H). 1 13 1313C NMR (101 MHz, DMSO) δ 170.68, 163.56, 162.78, 161.38, 155.72, 155.69, 152.38, 150.29, 148.89, 143.40, 138.67, 138.36, 137.23, 135.68, 135.42, 132.81, 131.18, 130.57, 130.28, 130.04, 129.26, 128.91, 116.20, 115.02, 114.41, 113.82, 112.27, 72.81, 70.47, 70.25, 69.99, 63.26, 60.69, 56.51, 54.04, 42.29, 36.24, 31.77, 31.23, 30.19, 24.14, 19.30, 18.54, 17.19, 14.24, 13.82, 12.92, 11.76.

[0132] Example 12

[0133] The JQ1-binding pyrimidine and pyrrole compound of the present invention has the chemical name of (S)-N-(2-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)ethyl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide, and the structural formula is as follows:

[0134]

[0135] Its preparation method includes the following steps:

[0136] (1) On the basis of Example 1, in step (1), N-tert-butoxycarbonyl-1,2-ethylenediamine is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the other conditions remain unchanged. Steps (2) and (3) are the same as those in Example 1, and the intermediate N-(2-aminoethyl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide is obtained, and the structural formula is as follows:

[0137]

[0138] (4) Add 100 mg (0.19 mmol) of the intermediate prepared in step (3), 71 mg (0.18 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid, 84 mg (0.20 mmol) of COMU, 54 mg (0.53 mmol) of NMI, and 2 ml of DMF solvent to the reaction flask in sequence. React at 25 - 35 °C for 2 hours. Monitor the reaction by TLC until it is completed (DCM:MeOH = 10:1). Concentrate under reduced pressure and purify by column chromatography (eluent DCM:MeOH = 30:1) to obtain 60 mg of the target product with a yield of 37.23%. The target compound is a white solid;

[0139] H NMR (400 MHz, DMSO-d6) δ 10.06 (s, 1H), 8.88 (s, 1H), 8.58 (s, 1H), 8.39 (s, 1H), 8.26 (d, J = 2.1 Hz, 1H), 8.16–8.08 (m, 1H), 7.46–7.35 (m, 6H), 6.98 (s, 1H), 5.43 (p, J = 9.2 Hz, 1H), 4.52 (dd, J = 8.0, 6.2 Hz, 1H), 3.26 (d, J = 7.0 Hz, 2H), 2.57 (s, 3H), 2.40–2.35 (m, 6H), 2.17 (d, J = 1.0 Hz, 3H), 1.90 (d, J = 6.0 Hz, 2H), 1.72 (s, 2H), 1.57–1.47 (m, 4H), 1.39 (s, 6H). 1 C NMR (101 MHz, DMSO) δ 170.74, 163.63, 162.33, 155.83, 155.74, 153.55, 152.53, 150.40, 147.38, 143.55, 139.28, 138.62, 137.31, 135.79, 135.58, 133.17, 132.86, 131.26, 130.74, 130.42, 130.16, 128.98, 125.93, 123.13, 114.95, 113.87, 112.95, 112.48, 110.02, 109.73, 107.45, 104.02, 56.75, 54.43, 38.81, 38.35, 30.33, 30.22, 26.94, 24.22, 14.54, 14.16, 13.23, 11.87. 13

[0140] Example 13 ​

[0141] The JQ1-binding pyrimido[2,3-d]pyrrole compound of the present invention has the chemical name of (S)-N-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propyl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide, and its structural formula is as follows:

[0142]

[0143] Its preparation method comprises the following steps:

[0144] (1) On the basis of Example 1, in step (1), N-Boc-1,3-propanediamine is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the remaining conditions remain unchanged. Steps (2) and (3) are the same as those in Example 1, to obtain the intermediate N-(3-aminopropyl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide, and its structural formula is as follows:

[0145]

[0146] (4) Add 100 mg (0.19 mmol) of the intermediate prepared in step (3), 70 mg (0.17 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid, 81 mg (0.19 mmol) of COMU, and 52 mg (0.52 mmol) of NMI into the reaction flask in sequence. Using 2 ml of DMF as the reaction solvent, react at 25 - 35 °C for 2 hours. Monitor the reaction by TLC until it ends (DCM:MeOH = 10:1), concentrate under reduced pressure, and purify by column chromatography (DCM:MeOH = 30:1) to obtain 87 mg of the target product, with a yield of 56.27%.

[0147] The target compound is a white solid; 1 ​1H NMR (400 MHz, DMSO-d6) δ 10.06 (s, 1H), 8.90 (s, 1H), 8.60 (t, J = 5.6 Hz, 1H), 8.34–8.26 (m, 1H), 8.15 (d, J = 1.4 Hz, 1H), 8.11 (s, 1H), 7.48–7.39 (m, 6H), 6.98 (s, 1H), 5.43 (p, J = 9.2 Hz, 1H), 4.53 (dd, J = 8.4, 5.8 Hz, 1H), 3.19 (td, J = 14.2, 13.3, 6.2 Hz, 4H), 2.60 (s, 3H), 2.39 (s, 6H), 2.19 (s, 3H), 1.92–1.88 (m, 2H), 1.74 (q, J = 7.2 Hz, 4H), 1.63–1.30 (m, 8H). 13 13C NMR (101 MHz, DMSO) δ 169.70, 163.06, 161.53, 155.20, 155.11, 152.90, 151.90, 149.81, 142.94, 138.66, 138.01, 136.76, 135.19, 134.98, 132.69, 132.27, 130.92, 130.67, 130.60, 130.13, 129.82, 129.56, 128.42, 125.20, 122.49, 114.35, 113.25, 112.28, 111.86, 109.08, 103.18, 56.10, 53.89, 37.72, 36.73, 36.32, 29.69, 29.12, 26.33, 23.65, 13.97, 13.56, 12.63, 11.28.

[0148] Example 14

[0149] The JQ1-binding pyrimido[2,3-d]pyrrole compound of the present invention has the chemical name of (S)-N-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butyl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide, and its structural formula is as follows:

[0150]

[0151] Its preparation method comprises the following steps:

[0152] (1) (1) Based on Example 1, step (1) uses N-tert-butyloxycarbonyl-1,4-butanediamine to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the other conditions remain unchanged. Steps (2) and (3) are the same as in Example 1 to obtain the intermediate N-(4-aminobutyl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide, the structural formula of which is as follows:

[0153]

[0154] (4) 100 mg (0.18 mmol) of the intermediate prepared in step (3), 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepine were added to the reaction flask in sequence. 68 mg (0.17 mmol) of 2-(6-yl)acetic acid, 79 mg (0.19 mmol) of COMU, 52 mg (0.51 mmol) of NMI and 2 ml of DMF were used as the reaction solvent. The reaction was carried out at 25-35° C. for 2 hours. The reaction was monitored by TLC until completion (DCM:MeOH=10:1). The product was concentrated under reduced pressure and purified by column chromatography (eluent DCM:MeOH=30:1) to obtain 41 mg of the target product in a yield of 26.11%.

[0155] The target compound is a white solid; 1 H NMR (400MHz, DMSO-d6) δ10.06(s,1H),8.89(s,1H),8.64(t,J=5.7Hz,1H),8.28(t,J=2.0Hz,1H),8.24 (t,J=5.7Hz,1H),8.15(d,J=1.4Hz,1H),8.11(d,J=1.8Hz,1H),7.49(t,J=1.7Hz,1H),7.45–7.37(m,4H ),6.97(s,1H),5.43(p,J=9.1Hz,1H),4.52(dd,J=8.4,5.7Hz,1H),3.33–3.01(m,6H),2.59(s,3H),2.4 9–2.22(m,6H),2.18(d,J=1.0Hz,3H),1.90(s,2H),1.76–1.71(m,2H),1.62–1.47(m,6H),1.39(s,2H). 1313C NMR (101 MHz, DMSO) δ 169.41, 163.00, 161.54, 155.19, 155.12, 152.84, 151.90, 149.80, 142.95, 138.66, 138.01, 136.75, 135.19, 134.98, 132.74, 132.26, 130.67, 130.07, 129.80, 129.56, 128.43, 125.33, 125.26, 122.83, 122.33, 114.35, 113.22, 112.54, 112.29, 111.87, 109.07, 103.16, 56.08, 53.87, 38.26, 37.65, 29.72, 26.76, 26.46, 26.33, 23.66, 22.64, 14.00, 13.57, 12.64, 11.28.

[0156] Example 15

[0157] The JQ1-binding pyrimidine and pyrrole compound of the present invention has the chemical name of (S)-N-(5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)pentyl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide, and the structural formula is as follows:

[0158]

[0159] Its preparation method comprises the following steps:

[0160] (1) On the basis of Example 1, in step (1), N-Boc-1,5-pentanediamine is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the remaining conditions remain unchanged. Steps (2) and (3) are the same as in Example 1, and the intermediate N-(5-aminopentyl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide is obtained, and the structural formula is as follows:

[0161]

[0162] (4) Add 100 mg (0.18 mmol) of the intermediate prepared in step (3), 66 mg (0.16 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid, 77 mg (0.18 mmol) of COMU, 50 mg (0.50 mmol) of NMI, and 2 ml of DMF as the reaction solvent to the reaction flask in sequence. React at 25 - 35 °C for 2 hours. Monitor the reaction by TLC until it is completed (DCM:MeOH = 10:1). Concentrate under reduced pressure and purify by column chromatography (eluent DCM:MeOH = 30:1) to obtain 79 mg of the target product, with a yield of 52.67%. The target compound is a white solid;

[0163] The target compound is a white solid; 1 H NMR (400 MHz, DMSO-d6) δ 10.05 (s, 1H), 8.86 (s, 1H), 8.61 (t, J = 5.6 Hz, 1H), 8.28 (s, 1H), 8.21 (t, J = 5.7 Hz, 1H), 8.12 (d, J = 15.3 Hz, 1H), 7.50–7.37 (m, 6H), 6.96 (s, 1H), 5.41 (d, J = 9.0 Hz, 1H), 4.51 (dd, J = 8.2, 5.8 Hz, 1H), 3.32–3.05 (m, 6H), 2.58 (s, 3H), 2.38 (s, 6H), 2.17 (s, 3H), 2.02–1.83 (m, 2H), 1.73 (t, J = 7.7 Hz, 2H), 1.66–1.43 (m, 10H), 1.38 (q, J = 7.9 Hz, 2H). 13 C NMR (101 MHz, DMSO) δ 169.82, 163.42, 161.94, 155.59, 155.53, 153.22, 152.30, 150.20, 143.38, 139.08, 138.42, 137.19, 135.64, 135.39, 133.21, 132.66, 131.33, 131.07, 130.48, 130.22, 129.97, 128.86, 125.62, 122.91, 117.06, 114.76, 113.64, 112.71, 112.27, 109.45, 103.57, 101.57, 56.49, 54.30, 39.21, 38.79, 38.08, 30.14, 29.30, 28.98, 24.15, 24.08, 14.41, 13.98, 13.05, 11.69.

[0164] Example 16

[0165] The JQ1-binding pyrimido[2,3-d]pyrrole compound of the present invention has the chemical name of (S)-N-(6-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butyl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide, and its structural formula is as follows:

[0166]

[0167] Its preparation method comprises the following steps:

[0168] (1) On the basis of Example 1, in step (1), N-Boc-1,6-hexanediamine is used to replace tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate, and the remaining conditions remain unchanged. Steps (2) and (3) are the same as those in Example 1, and the intermediate N-(6-aminohexyl)-7-cyclopentyl-2-((3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide is obtained, and its structural formula is as follows:

[0169]

[0170] (4) 100 mg (0.17 mmol) of the intermediate prepared in step (3), 64 mg (0.15 mmol) of 2-((6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-3a,10a-dihydro-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid, 75 mg (0.17 mmol) of COMU, 49 mg (0.49 mmol) of NMI are successively added to the reaction flask, 2 ml of DMF is used as the reaction solvent, and the reaction is carried out at 25-35 °C for 2 hours. The reaction is monitored by TLC until it ends (DCM:MeOH = 10:1), concentrated under reduced pressure, and purified by column chromatography (eluent DCM:MeOH = 30:1) to obtain 43 mg of the target product, with a yield of 30.13%. The target compound is a yellowish-white solid;

[0171] The target compound is a yellowish-white solid; 11H NMR (400 MHz, DMSO-d6) δ 10.05 (s, 1H), 8.88 (s, 1H), 8.59 (t, J = 5.7 Hz, 1H), 8.28 (t, J = 2.0 Hz, 1H), 8.19–8.12 (m, 1H), 8.11 (d, J = 1.9 Hz, 1H), 7.50–7.37 (m, 6H), 6.93 (s, 1H), 5.42 (t, J = 9.0 Hz, 1H), 4.52 (dd, J = 8.0, 6.2 Hz, 1H), 3.33–3.03 (m, 6H), 2.58 (s, 3H), 2.47–2.28 (m, 6H), 2.18 (d, J = 1.1 Hz, 3H), 1.91 (d, J = 8.6 Hz, 2H), 1.74 (s, 2H), 1.59–1.44 (m, 6H), 1.34 (dt, J = 6.7, 3.0 Hz, 6H). 13 13C NMR (101 MHz, DMSO) δ 169.34, 162.97, 161.47, 155.17, 155.10, 152.78, 151.88, 149.75, 142.95, 138.66, 137.99, 136.74, 136.37, 135.22, 134.95, 132.82, 132.24, 130.63, 130.06, 129.77, 129.53, 128.41, 125.24, 122.50, 114.32, 113.17, 112.22, 111.85, 109.03, 103.08, 56.05, 53.90, 38.75, 38.31, 38.22, 37.66, 29.72, 29.21, 28.92, 26.32, 26.09, 25.99, 23.65, 13.97, 13.56, 12.60, 11.24.

[0172] Biological activity evaluation

[0173] (1) Activity test of the compound against cancer cells

[0174] To investigate the effect of the compound on the activity of cancer cells, MTT assay was used to detect cytotoxicity.

[0175] Test method:

[0176] Cell culture: Human cancer cell lines Hela and MDA-MB-231 cells were both purchased from the National Biomedical Laboratory in Beijing and cultured in DMEM (KGM12800-500) or MEM medium (KGM41500-500) containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, and cultured in a constant temperature incubator (Thermo Fisher Scientific, BB150) with 5% CO2 at 37°C. When the cell confluence reached 70%-80%, 0.25% trypsin was added for digestion, resuspension, and culture. Cells in the logarithmic growth phase and with good growth status were selected for the study.

[0177] Methyl thiazolyl tetrazolium (MTT) was used for the determination of cell viability. The hemocytometer method was used for cell counting, and in all experiments, the cell viability was greater than 95%. MDA-MB-231 and Hela cells were seeded at 1×10 4 cells per milliliter in 96-well plates. Different concentrations of the drug (0 - 50 μM) dissolved in 100 μL of medium (containing 1% FBS) were added to each well and incubated for 24 h respectively. Using a plate centrifuge (5 min, 2000 rpm), the supernatant was discarded, 10 μL of MTT (5 mg / mL) solution was added to each well, incubated at 37°C for 4 h, and the plate was centrifuged to discard the supernatant. 100 μL of DMSO was added to each well and placed on a shaker for 10 min to fully dissolve the formazan crystals. The absorbance was measured at a wavelength of 570 nm using a microplate reader (BioTek, USA). Cytotoxicity was determined compared with the control group (DMSO). The concentration of the drug that induced 50% cell growth inhibition (IC 50 ) was determined by non-linear regression using the curve fitting algorithm of GraphPad Prism 9 (GraphPad software, La Jolla, CA, USA). As shown in Table 1, the effects of the compound on the activities of different tumor cells were shown, and the half inhibitory concentration IC 50 (48 h) of the drug on each cell line was calculated.

[0178] Table 1 Inhibitory activities of compounds against different tumor cells

[0179] Example Hela (μM) MDA-MB-231 (μM) Example 1 11.3±0.63 6.07±0.11 Example 2 0.78±0.10 3.66±0.31 Example 3 6.85±0.75 0.56±0.07 Example 4 11.88±0.86 3.97±0.30 Example 5 6.98±1.10 1.61±0.03 Example 6 2.90±0.80 0.84±0.15 Example 7 22.58±1.01 4.89±0.52 Example 8 2.59±1.22 2.18±0.43 Example 9 4.01±1.01 1.23±0.28 Example 10 7.57±1.51 3.9±0.56 Example 11 21.96±4.95 5.79±1.65 Example 12 0.30±0.03 2.47±0.04 Example 13 2.61±0.71 0.22±0.02 Example 14 3.66±0.33 5.77±0.37 Example 15 0.20±0.02 7.96±1.63 Example 16 39.06±5.49 9.85±1.51 (+)-JQ-1 42.88±0.78 23.93±1.33

[0180] The results in Table 1 showed that the compound had a significant inhibitory effect on HELA and MDA-MB-231 tumor cells, some reaching below 1 μM, which was significantly better than the compound (+)-JQ-1, indicating that the compound had good activity against tumor cells and had great research value.

[0181] (2) Protein degradation experiment

[0182] Experimental purpose: To observe the degradation effect of the compound on BRD4 protein.

[0183] Test method: Cultivate MDA-MB-231 cells in vitro. Seed the cells in a 6-well plate. When the cells grow to about 80%, add different concentrations (1.25 μM to 20 μM) of the compound of Example 13. After acting for 4 hours, collect the cells, extract the protein, and perform Western Blotting to detect the BRD4 protein level.

[0184] The experimental results are shown in Figure 2 , the compound of the present invention can effectively degrade the target protein BRD4 in tumor cells and shows a concentration-dependent manner.

Claims

1. A compound of JQ1 combined with pyrimidine and pyrrole, characterized in that, The chemical structural formula is as follows: Among them, Linker is Among them, R1 is H or a three-membered ring; R2 is a 4- to 6-membered nitrogen-containing heterocycle and the nitrogen atom is connected to the carbonyl group, n1 is 0 or 1; a, b, and c are selected from 4- to 6-membered nitrogen-containing heterocycles and the nitrogen atom is connected to the carbonyl group; n2 is 1 to 10.

2. The JQ1 combined with pyrimidine and pyrrole compound according to claim 1, characterized in that, The said is 3. The compound of JQ1 combined with pyrimidine and pyrrole according to claim 1, characterized in that, The said is 4. The JQ1 combined with pyrimidine and pyrrole compound according to claim 1, characterized in that, The said is 5. The JQ1 combined with pyrimidine and pyrrole compound according to claim 1, wherein The structural formula of the compound of JQ1 combined with pyrimidine and pyrrole is as follows:

6. A method for preparing the JQ1-binding pyrimidine and pyrrole compound according to claim 1, characterized in that, It includes the following steps: (1) Compound 1 and the Boc-protected linker undergo an amidation reaction under alkaline conditions to obtain intermediate 2; (2) Intermediate 2 and Compound 3 undergo a Buchwald-Hartwig coupling reaction to obtain intermediate 4; (3) Intermediate 4 is hydrolyzed to obtain intermediate 5; (4) Intermediate 5 and Compound 6 undergo an amidation reaction under alkaline conditions to obtain the target product; The synthesis route is as follows:

7. The preparation method of the JQ1 combined with pyrimidine and pyrrole compound according to claim 3, characterized in that, The temperature of the amidation reaction is 0 - 30 °C.

8. The method for preparing the JQ1 combined with pyrimidine and pyrrole compound according to claim 3, characterized in that, In step (2), the solvent for the Buchwald-Hartwig coupling reaction is dimethyl sulfoxide, N,N-dimethylformamide, 1,4-dioxane, preferably 1,4-dioxane; the catalyst for the reaction is palladium acetate, palladium chloride or [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium; the catalyst ligand is 1,1'-binaphthalene-2,2'-bis(diphenylphosphine) or 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthrene; the reaction temperature is 100 - 150 °C; the reaction inert condition is nitrogen or argon protection.

9. A tautomer, optical isomer, deuterated compound, nitrogen oxide, solvate, pharmaceutically acceptable salt or prodrug of the compound of JQ1 combined with pyrimidine and pyrrole as claimed in claims 1 - 5.

10. Use of the compound of JQ1 combined with pyrimidine and pyrrole as claimed in any one of claims 1 - 5 in the preparation of a drug for treating tumors.

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