A class of biaromatic BET inhibitors, their synthesis methods and applications

Abstract

This invention discloses a class of biaromatic ring-based BET inhibitors, their synthesis methods, and uses. The compounds with biaromatic ring structures provided by this invention exhibit novel structures and excellent inhibitory effects on BET proteins. Those skilled in the art know that BET proteins are targets for tumor or inflammation treatment; therefore, the compounds with biaromatic ring structures provided by this invention can be used alone to prepare antitumor or anti-inflammatory drugs, or they can be combined with other antitumor components to prepare antitumor drugs, such as PARP inhibitors, CDK4 / 6 inhibitors, ATR inhibitors, PD-1 antibodies or small molecule inhibitors, PD-L1 antibodies or small molecule inhibitors, EZH2 inhibitors, or AR inhibitors.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to a class of biaromatic BET inhibitors, their synthesis methods, and uses. Background Technology

[0002] The bromo- and superterminal (BET) protein family of epigenetic targets is an extremely important target in the treatment of tumors and inflammation. The BET protein family includes four subtypes: BRD2, BRD3, BRD4, and BRDT. All BET family proteins contain two highly conserved N-terminal tandem bromodomains, BD1 and BD2. Taking BRD4 as an example, the first N-terminal bromodomain of BRD4 is represented as BRD4(1), and the second N-terminal bromodomain of BRD4 is represented as BRD4(2). BET proteins recognize acetylated lysine residues of histones and other proteins through bromodomains, playing an important role in regulating gene expression and controlling cell growth. Many human diseases are closely related to the overexpression of BET proteins, such as tumors, inflammatory diseases, autoimmune diseases, and viral infections. The development of BET protein inhibitors has great value in the fields of anti-tumor and anti-inflammatory treatments and has therefore been widely studied by major pharmaceutical companies and research institutions.

[0003] This invention is proposed to provide a more promising BET inhibitor. Summary of the Invention

[0004] The first objective of this invention is to provide a class of compounds with a biaromatic ring structure, the second objective is to provide a method for preparing such compounds with a biaromatic ring structure, and the third objective is to provide the uses of such compounds with a biaromatic ring structure.

[0005] The above-mentioned objective of this invention is achieved through the following technical solution:

[0006] A class of compounds with a biaromatic ring structure, namely the compound shown in Formula I or a pharmaceutically acceptable salt thereof:

[0007]

[0008] in:

[0009] Ring A is selected from phenyl, 5-10 aryl heterogroups; R a Independently selected from hydrogen, halogen, C 1-5 Alkyl, C 3-7 cycloalkyl, C 1-3 Alkoxy group; n = 1-5; ring B selected from 5-10 aryl heterogroups, R b Independently selected from hydrogen, halogen, C 1-5 Alkyl, C 3-7 cycloalkyl, C 1-3 Alkyl group; m = 1-5;

[0010] R1, R2, R3, and R4 are independently selected from hydrogen, halogens, and C. 1-5 Alkyl, C 1-3 Alkyl groups, -S(O) p R * -NRR * S(O) p R * -NRR * S(O) p NRR * -C(O)OR * -C(O)R * -OC(O)R * -OC(O)NRR * -NRC(O)R * -NRC(O)NRR * -C(O)NRR * -NRR * -OR * -SR * ;R and R * Independently selected from hydrogen and C 1-5 Alkyl, C 2-6 alkenyl, C 2-6 alkenyl, C 2-6 Alkyne group, or R and R * The nitrogen atoms attached to them form 4-7 membered heterocyclic alkyl groups; p = 1 or 2.

[0011] Preferably, the pharmaceutically acceptable salt is an acid addition salt of a compound of formula I, wherein the acid used for salt formation includes hydrogen chloride, sulfuric acid, hydrogen bromide, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid.

[0012] A method for synthesizing the above-mentioned compounds, the structures of which are shown in targets 1 to 14, and the synthetic route is as follows:

[0013]

[0014] A method for synthesizing the above-mentioned compounds, the structures of which are shown in targets 15 to 20, and the synthetic route are as follows:

[0015]

[0016] A method for synthesizing the above-mentioned compounds, the structures of which are shown in targets 21 to 24, and the synthetic route are as follows:

[0017]

[0018] A method for synthesizing the above-mentioned compounds, the structures of which are shown in targets 25 to 30, and the synthetic route are as follows:

[0019]

[0020] The above compounds are used in the preparation of BET inhibitor drugs.

[0021] The above compounds are used in the preparation of antitumor drugs or anti-inflammatory drugs.

[0022] Preferably, the antitumor drug further contains other antitumor components, such as PARP inhibitors, CDK4 / 6 inhibitors, ATR inhibitors, PD-1 antibodies or small molecule inhibitors, PD-L1 antibodies or small molecule inhibitors, EZH2 inhibitors or AR inhibitors.

[0023] Beneficial effects:

[0024] The compounds with biaromatic ring structures provided by this invention have novel structures and exhibit excellent inhibitory effects on BET protein. Those skilled in the art know that BET protein is a target for tumor or inflammation treatment; therefore, the compounds with biaromatic ring structures provided by this invention can be used alone to prepare antitumor or anti-inflammatory drugs, or they can be combined with other antitumor components to prepare antitumor drugs, such as PARP inhibitors, CDK4 / 6 inhibitors, ATR inhibitors, PD-1 antibodies or small molecule inhibitors, PD-L1 antibodies or small molecule inhibitors, EZH2 inhibitors, or AR inhibitors. Attached Figure Description

[0025] Figure 1 Example 26 illustrates the inhibitory effect of combined use with a PARP inhibitor on the proliferation of breast and ovarian cancer cells. Detailed Implementation

[0026] The substantive content of the present invention is described in detail below with reference to embodiments, but this is not intended to limit the scope of protection of the present invention. Experimental methods in the following embodiments that do not specify specific conditions were implemented according to existing industry methods and conditions.

[0027] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰ ohms. -6 The unit (ppm) is given. NMR measurements were performed using a Bruker AVANCE-300 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard.

[0028] MS measurements were performed using an MS liquid chromatography-mass spectrometry system (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS). HRMS used an Agilent 6230.

[0029] The silica gel plates used for thin-layer chromatography are Qingdao GF254 silica gel plates. The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15 mm to 0.2 mm, and the diameter of the silica gel plates used for TLC separation and purification products is 0.4 mm to 0.5 mm. Other starting materials disclosed in this invention can be synthesized according to methods known in the art or derived from commercially available products.

[0030] Unless otherwise specified in the examples, the reactions can be carried out under an argon or nitrogen atmosphere.

[0031] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20℃~30℃.

[0032] Synthesis Route 1:

[0033] Examples 1-14 were synthesized according to Scheme 1.

[0034] Commercial raw materials 1-13 were respectively placed in a Shrek tube with 15, cesium carbonate, and a [1,1-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (Pd(dppf)Cl2·CH2Cl2) and subjected to a Suzuki coupling reaction under heating at 100°C and argon protection to obtain Examples 1-13. Commercial raw material 16 and N,N′-carbonyldiimidazole 17 were heated in tetrahydrofuran to obtain intermediate 14. 14 and 15 were subjected to a Suzuki coupling reaction under the same conditions to obtain Example 14.

[0035]

[0036] Scheme 1.Reagents and conditions: (a) Cs2CO3, Pd(dppf)Cl2·CH2Cl2, 1,4-Dioxane, H2O, 100℃, 3h; (b) THF, 80℃, 4h.

[0037] Example 1:

[0038] Synthesis of 5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (Example 1)

[0039] 5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (Example 1)

[0040]

[0041] Conventional Route A: 5-Bromo-1H-pyrrolo[2,3-b]pyridine 1 (300 mg, 1.52 mmol), 3,4,5-trimethoxyphenylboronic acid 15 (419.64 mg, 1.98 mmol), cesium carbonate (992.17 mg, 3.05 mmol), and [1,1-bis(diphenylphosphine)ferrocene]palladium dichloromethane complex (124.38 mg, 0.152 mmol) were added to a Shrek tube, followed by 9 mL of dioxane and 3 mL of water as the reaction solvent. The air in the Shrek tube was completely replaced with argon, and the reaction was carried out at 100 °C for 3 h. The reaction was monitored by TLC until it was completely finished. After the reaction was cooled to room temperature, equal volumes of ethyl acetate and water were added for three hydration extractions. The organic layers were combined and concentrated under low pressure to remove the solvent. The residue was subjected to column chromatography to obtain the target product Example 1 (317 mg, yield: 73.2%). 1 HNMR (300MHz, DMSO-d6) δ11.74(s,1H),8.58(d,J=2.2Hz,1H),8.26(dd,J=2.2,0.7Hz,1H),7. 54(dd,J=3.5,2.5Hz,1H),6.99(s,2H),6.53(dd,J=3.4,1.9Hz,1H),3.91(s,6H),3.73(s,3H).

[0042] Example 2:

[0043] Synthesis of 6-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (Example 2)

[0044] 6-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (Example 2)

[0045]

[0046] Following conventional route A, using 6-bromo-1H-pyrrolo[2,3-b]pyridine 2 (300 mg, 1.52 mmol) as the reactant, the target product Example 2 (297 mg, yield: 68.75%) was obtained. 1 H NMR(300MHz,)δ11.78(s,1H),8.05(d,J=8.3Hz,1H),7.73(d,J=8.3Hz,1H),7.50(dd,J =3.5,2.5Hz,1H),7.42(s,2H),6.49(dd,J=3.4,1.8Hz,1H),3.92(s,6H),3.75(s,3H).

[0047] Example 3:

[0048] Synthesis of 5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (Example 3)

[0049] 5-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (Example 3)

[0050]

[0051] Following conventional route A, using 4-bromo-1H-pyrrolo[2,3-b]pyridine 3 (300 mg, 1.52 mmol) as the reactant, the target product Example 3 (327 mg, yield: 75.7%) was obtained. 1 H NMR(300MHz,)δ11.82(s,1H),8.30(d,J=5.0Hz,1H),7.57(t,J=3.0Hz,1H),7.26(d ,J=5.0Hz,1H),7.05(s,2H),6.72(dd,J=3.5,1.8Hz,1H),3.91(s,6H),3.77(s,3H).

[0052] Example 4:

[0053] Synthesis of 3-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (Example 4)

[0054] 3-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (Example 4)

[0055]

[0056] Following conventional route A, using 4-bromo-1H-pyrrolo[2,3-b]pyridine 4 (300 mg, 1.52 mmol) as the reactant, the target product Example 4 (288 mg, yield: 66.7%) was obtained. 1 H NMR(300MHz,)δ11.92(s,1H),8.42–8.24(m,2H),7.90(d,J=2.7Hz,1H),7.19(dd,J=8.0,4.7Hz,1H),6.97(s,2H),3.91(s,6H),3.72(s,3H).

[0057] Example 5:

[0058] Synthesis of 4-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-c]pyridine (Example 5)

[0059] 4-(3,4,5-trimethoxyphenyl)-1H-pyrrolo[2,3-c]pyridine (Example 5)

[0060]

[0061] Following conventional route A, using 4-bromo-1H-pyrrolo[2,3-c]pyridine 5 (300 mg, 1.52 mmol) as the reactant, the target product Example 5 (293 mg, yield: 67.8%) was obtained. 1 H NMR (300MHz, DMSO-d6) δ11.80(s,1H),8.76(s,1H),8.28(s,1H),7.70(t,J=2.8Hz,1H),6.99(s,2H),6.83–6.65(m,1H),3.89(s,6H),3.75(s,3H).

[0062] Example 6:

[0063] Synthesis of 4-(3,4,5-trimethoxyphenyl)-1H-benzo[d]imidazole (Example 6)

[0064] 4-(3,4,5-trimethoxyphenyl)-1H-benzo[d]imidazole (Example 6)

[0065]

[0066] Following conventional route A, using 4-bromo-1H-benzo[d]imidazole 6 (300 mg, 1.52 mmol) as the reactant, the target product Example 6 (339 mg, yield: 78.4%) was obtained. 1 HNMR (300MHz, DMSO-d6) δ12.63 (s, 1H), 8.29 (d, J = 23.4Hz, 1H), 7.72–7.26 (m, 5H), 6.92 (s, 1H), 3.76 (s, 3H).

[0067] Example 7:

[0068] Synthesis of 4-(3,4,5-trimethoxyphenyl)-1H-indazole (Example 7)

[0069] 4-(3,4,5-trimethoxyphenyl)-1H-indazole (Example 7)

[0070]

[0071] Following conventional route A, using 4-bromo-1H-indazole 7 (300 mg, 1.52 mmol) as the reactant, the target product Example 7 (323 mg, yield: 74.7%) was obtained. 1 H NMR (300MHz, DMSO-d6) δ12.63 (s, 1H), 8.29 (d, J = 23.4Hz, 1H), 7.72–7.26 (m, 5H), 6.92 (s, 1H), 3.76 (s, 3H).

[0072] Example 8:

[0073] Synthesis of 4-(3,4,5-trimethoxyphenyl)benzo[d]isoxazole (Example 8)

[0074] 4-(3,4,5-trimethoxyphenyl)benzo[d]isoxazole (Example 8)

[0075]

[0076] Following conventional route A, using 4-bromobenzo[d]isoxazole 8 (250 mg, 1.26 mmol) as the reactant, the target product Example 8 (216 mg, yield: 76.1%) was obtained. 1 H NMR (300MHz, DMSO-d6) δ11.21(d,J=4.5Hz,1H),7.57(td,J=7.9,2.7Hz,1H),7.14–6.82(m,4H),3.88(s,6H),3.81–3.74(m,3H).

[0077] Example 9:

[0078] Synthesis of 8-(3,4,5-trimethoxyphenyl)-[1,2,4]triazolo[4,3-a]pyridine (Example 9)

[0079] 8-(3,4,5-trimethoxyphenyl)-[1,2,4]triazolo[4,3-a]pyridine (Example 9)

[0080]

[0081] Following conventional route A, using 8-bromo-[1,2,4]triazolo[4,3-a]pyridine 9 (250 mg, 1.26 mmol) as the reactant, the target product Example 9 (178 mg, yield: 62.4%) was obtained. 1H NMR(300MHz,DMSO-d6)δ9.40(d,J=3.5Hz,1H),8.70–8.52(m,1H),7.77(dd,J=7 .2,3.6Hz,1H),7.63(d,J=3.7Hz,2H),7.19–7.03(m,1H),3.91(d,J=3.5Hz,6H).

[0082] Example 10:

[0083] Synthesis of 5-(3,4,5-trimethoxyphenyl)-[1,2,4]triazolo[4,3-a]pyridine (Example 10)

[0084] 5-(3,4,5-trimethoxyphenyl)-[1,2,4]triazolo[4,3-a]pyridine (Example 10)

[0085]

[0086] Following conventional route A, using 5-bromo-[1,2,4]triazolo[4,3-a]pyridine 10 (250 mg, 1.26 mmol) as the reactant, the target product Example 10 (165 mg, yield: 57.8%) was obtained. 1 HNMR (300MHz, DMSO-d6) δ9.39(s,1H),7.82(d,J=9.2Hz,1H),7.49(dd,J=9.2,6.8Hz,1H),7.11(s,2H),7.09–7.05(m,1H),3.90(s,6H),3.79(s,3H).

[0087] Example 11:

[0088] Synthesis of 4-(3,4,5-trimethoxyphenyl)-1H-pyrazolo[3,4-b]pyridine (Example 11)

[0089] 4-(3,4,5-trimethoxyphenyl)-1H-pyrazolo[3,4-b]pyridine (Example 11)

[0090]

[0091] Following conventional route A, using 4-bromo-1H-pyrazolo[3,4-b]pyridine 11 (200 mg, 1.01 mmol) as the reactant, the target product Example 11 (152 mg, yield: 53.1%) was obtained. 1HNMR (300MHz, DMSO-d6) δ13.81(s,1H),8.60(d,J=4.8Hz,1H),8.45(s,1H),7.44(d,J=4.8Hz,1H),7.14(s,2H),3.79(s,3H).

[0092] Example 12:

[0093] Synthesis of 7-(3,4,5-trimethoxyphenyl)-3H-imidazo[4,5-b]pyridine (Example 12)

[0094] 7-(3,4,5-trimethoxyphenyl)-3H-imidazo[4,5-b]pyridine (Example 12)

[0095]

[0096] Following conventional route A, using 7-chloro-3H-imidazo[4,5-b]pyridine 12 (200 mg, 1.3 mmol) as the reactant, the target product Example 12 (121 mg, yield: 42.4%) was obtained. 1 HNMR (300MHz, DMSO-d6) δ13.25(s,1H),8.53(s,1H),8.40(d,J=5.1Hz,1H),7.75(s,2H),7.65(d,J=5.3Hz,1H),3.92(d,J=3.3Hz,6H),3.77(s,3H).

[0097] Example 13:

[0098] Synthesis of 4-(3,4,5-trimethoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidine (Example 13)

[0099] 4-(3,4,5-trimethoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidine (Example 13)

[0100]

[0101] Following conventional route A, using 4-bromo-7H-pyrrolo[2,3-d]pyrimidine 13 (200 mg, 1.01 mmol) as the reactant, the target product Example 13 (169 mg, yield: 59.3%) was obtained. 1HNMR (300MHz, DMSO-d6) δ12.28(s,1H),8.85(s,1H),7.68(dd,J=3.6,2.3Hz,1H),7.46(s,2H),6.96(dd,J=3.7,1.7Hz,1H),3.94(s,6H),3.79(s,3H).

[0102] Example 14:

[0103] Synthesis of 4-(3,4,5-trimethoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-b]pyridin-2-one (Example 14)

[0104] 4-(3,4,5-trimethoxyphenyl)-1,3-dihydro-2H-benzo[d]imidazol-2-one (Example 14)

[0105]

[0106] Following conventional route A, using 7-chloro-1,3-dihydro-2H-imidazo[4,5-b]pyridin-2-one 14 (283 mg, 1.67 mmol) as the reactant, the target product Example 14 (293 mg, yield: 53.3%) was obtained. 1 H NMR (300MHz, DMSO-d6) δ11.47(s,1H),11.08(s,1H),7.95(d,J=5.5Hz,1H),7.11(d,J=5.5Hz,1H),6.87(s,2H),3.91(s,6H),3.74(s,3H).

[0107] Compound 14 was synthesized by the following method:

[0108] Synthesis of 7-chloro-1,3-dihydro-2H-imidazo[4,5-b]pyridin-2-one (14)

[0109] 7-bromo-1,3-dihydro-2H-imidazo[4,5-b]pyridin-2-one(14)

[0110]

[0111] 4-Chloropyridine-2,3-diamine 16 (300 mg, 2.09 mmol) and N,N'-carbonyldiimidazole 17 (677 mg, 4.18 mmol) were added to a round-bottom flask containing 10 mL of tetrahydrofuran. The mixture was heated at 80 °C for 4 h, and the reaction was confirmed to be complete by TLC. The tetrahydrofuran was removed by concentration under reduced pressure, and the residue was dissolved in 10 mL of dichloromethane. A large amount of red powder precipitated out, and after filtration, 14 (283 mg, yield: 79.9%) was obtained.

[0112] Synthesis Route 2:

[0113] Examples 15–20 were synthesized according to Scheme 2. Commercially available compounds 19a–19f were reacted with 1-bromo-3-iodobenzene 18 in a Shrek tube under Scheme 2 conditions, using Pd(dppf)Cl2·CH2Cl2 as a catalyst, cesium carbonate as a base, and dioxane and H2O (3:1) as a solvent, at 100°C under argon protection, via Suzuki coupling to yield intermediates 20a–20f. Intermediate 20f reacted with bromomethylcyclopropane 24 in the presence of cesium carbonate via affinity substitution to yield intermediate 20g. Intermediates 20a–20e and 20g reacted with 21 under Pd(dppf)Cl2·CH2Cl2 and potassium tert-butoxide catalysis, using anhydrous dioxane as a solvent, under argon protection and heating conditions, to yield borate intermediates 22a–22e and 22g. This step required strictly anhydrous and oxygen-free conditions. Examples 15 to 20 were obtained by reacting 22a to 22e and 22g with 23 in a mixture of Pd(dppf)Cl2·CH2Cl2 as a catalyst, cesium carbonate as a base, and dioxane and H2O (3:1) as a solvent, and heating at 100°C for 7 hours under argon protection.

[0114]

[0115] Scheme 2.Reagents and conditions: (a) Cs2CO3, Pd(dppf)Cl2·CH2Cl2, 1,4-Dioxane, H2O, 100℃, 3h; (b) AcOK, Pd(dppf)Cl2·CH2Cl2, dry 1,4-Dioxane, 100℃, 3h; (c) Cs2CO3, Pd(dppf)Cl2·CH2Cl2, 1,4-Dioxane, H2O, 100℃, 7h; (d) Cs2CO3, DMF, rt, 5h.

[0116] Synthesis of 4-(3-bromophenyl)-3,5-dimethylisoxazole (20a)

[0117] 4-(3-bromophenyl)-3,5-dimethylisoxazole(20a)

[0118]

[0119] Conventional synthetic route B: 1-Bromo-3-iodobenzene 18 (500 mg, 1.77 mmol), 3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)isoxazole 19a (512.54 mg, 2.30 mmol), cesium carbonate (1.15 g, 3.53 mmol), and [1,1-bis(diphenylphosphine)ferrocene]palladium dichloromethane dichloride complex (143.90 mg, 0.177 mmol) were added to a Shrek tube, followed by 9 mL of dioxane and 3 mL of water as the reaction solvent. The air in the Shrek tube was completely replaced with argon, and the reaction was carried out at 100 °C for 3 h. TLC monitoring showed that the reaction was complete. After the reaction was cooled to room temperature, equal volumes of ethyl acetate and water were added for three extractions. The organic layers were combined and the solvent was removed by low-pressure concentration. The residue was subjected to column chromatography to obtain the target product 20a (400 mg, yield: 89.8%). 1 H NMR (300MHz, Chloroform-d) δ7.73(m,1H),7.61(d,J=7.5,1H),7.43(d,J=7.5Hz,1H),7.41(t,J=7.5Hz,1H),2.87(s,3H),2.66(s,3H).

[0120] Synthesis of 4-(3-bromophenyl)-1,3,5-trimethyl-1H-pyrazole (20b)

[0121] 4-(3-bromophenyl)-1,3,5-trimethyl-1H-pyrazole(20b)

[0122]

[0123] Following conventional route B, using 1,3,5-trimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-1H-pyrazole 19b (533 mg, 2.30 mmol) as the reactant, the target product 20b (413 mg, yield: 88.13%) was obtained. 1HNMR(300MHz,Chloroform-d)δ7.70(t,J=2.0Hz,1H),7.61(dt,J=7.5,2.0Hz,1H),7.4 3(dt,J=7.5,2.1Hz,1H),7.34(t,J=7.5Hz,1H),3.92(s,3H),2.61(s,3H),2.37(s,3H).

[0124] Synthesis of 5-(3-bromophenyl)-1,4-dimethyl-1H-pyrazole (20c)

[0125] 5-(3-bromophenyl)-1,4-dimethyl-1H-pyrazole(20c)

[0126]

[0127] Following conventional route B, using 1,4-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1,4-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole)19c (510 mg, 2.30 mmol) as the reactant, the target product 20c (398 mg, yield: 89.67%) was obtained. 1 HNMR(300MHz,Chloroform-d)δ7.70(t,J=2.0Hz,1H),7.61(dt,J=7.3,2.1Hz,1H),7.4 2(dt,J=7.5,2.0Hz,1H),7.39(s,1H),7.34(t,J=7.5Hz,1H),3.85(s,3H),2.08(s,3H).

[0128] Synthesis of 3'-bromo-2,6-dimethyl-1,1'-biphenyl (20d)

[0129] 3'-bromo-2,6-dimethyl-1,1'-biphenyl(20d)

[0130]

[0131] Following conventional route B, using 2-(2,6-dimethylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxoborane 19d (533 mg, 2.30 mmol) as the reactant, the target product 20d (430 mg, yield: 93.16%) was obtained. 1H NMR (300MHz, Chloroform-d) δ7.70–7.59(m,2H),7.40(dt,J=7.6,2.0Hz,1H),7.31(dt,J=15.4,7.5Hz,2H),7.16(d,J=7.5Hz,2H),2.27(s,6H).

[0132] Synthesis of 8-(3-bromophenyl)quinoline (20e)

[0133] 8-(3-bromophenyl)quinoline(20e)

[0134]

[0135] Following conventional route B, using 8-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)quinoline 19e (586 mg, 2.30 mmol) as the reactant, the target product 20e (410 mg, yield: 81.64%) was obtained. 1 H NMR (300MHz, Chloroform-d) δ9.00 (dd, J=7.5, 1.5Hz, 1H), 8.31 (dt, J=7.5, 1.5Hz, 1H), 7.89 (dt, J=7.6, 1. 5Hz,1H),7.86–7.79(m,2H),7.67–7.57(m,2H),7.55(dt,J=7.5,2.0Hz,1H),7.38(dt,J=14.7,7.5Hz,2H).

[0136] Synthesis of 4-(3-bromophenyl)-3,5-dimethyl-1H-pyrazole (20f)

[0137] 4-(3-bromophenyl)-3,5-dimethyl-1H-pyrazole(20f)

[0138]

[0139] Following conventional route B, using 3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-1H-pyrazole 19f (510.28 mg, 2.30 mmol) as the reactant, the target product 20f (356 mg, yield: 80.21%) was obtained.

[0140] Synthesis of 4-(3-bromophenyl)-1-(cyclopropylmethyl)-3,5-dimethyl-1H-pyrazole (20g)

[0141] 4-(3-bromophenyl)-1-(cyclopropylmethyl)-3,5-dimethyl-1H-pyrazole(20g)

[0142]

[0143] At room temperature, 4-(3-bromophenyl)-3,5-dimethyl-1H-pyrazole 20f (500 mg, 1.99 mmol), (bromomethyl)cyclopropane 24 (403.19 mg, 2.99 mmol), and cesium carbonate (1.30 g, 3.98 mmol) were added to a round-bottom flask, followed by 15 mL of LDM as the reaction solvent. After stirring at room temperature for 4 h, the reaction was confirmed to be complete by TLC. The mixture was extracted three times with equal volumes of ethyl acetate and water. The combined organic layers were extracted again with saturated sodium chloride solution. The organic layer was dried over anhydrous sodium sulfate for 1 h and then concentrated to remove the solvent. Column chromatography yielded 20 g (467 mg, 76.9%) of a white oily substance.

[0144] Synthesis of 3,5-dimethyl-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoborhecyclopentan-2-yl)phenyl)isoxazole (22a)

[0145] 3,5-dimethyl-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)isoxazole(22a)

[0146]

[0147] Conventional synthetic route C: 4-(3-bromophenyl)-3,5-dimethylisoxazole 20a (334 mg, 1.33 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-di(1,3,2-dioxaborane)21 (438.07 mg, 1.73 mmol), potassium acetate (260.47 mg, 2.65 mmol), and [1,1-bis(diphenylphosphine)ferrocene]palladium dichloromethane dichloride complex (108.79 mg, 0.13 mmol) were added to a Shrek tube, followed by 10 mL of anhydrous dioxane as the reaction solvent. The air in the Shrek tube was completely replaced with argon, and the reaction was carried out at 100 °C for 3 h. TLC monitoring showed that the reaction was complete. After the reaction was cooled to room temperature, equal volumes of ethyl acetate and water were added for three hydration extractions. The organic layers were combined and concentrated under low pressure to remove the solvent, yielding a residue of 320 mg. Without further purification, it is directly used in the next step.

[0148] Synthesis of 1,3,5-trimethyl-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)phenyl)-1H-pyrazole (22b)

[0149] 1,3,5-trimethyl-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazole(22b)

[0150]

[0151] Following conventional route C, using 4-(3-bromophenyl)-1,3,5-trimethyl-1H-pyrazole 20b (400 mg, 1.51 mmol) as the reactant, the target product 22b (390 mg, yield: 82.8%) was obtained.

[0152] Synthesis of 1,4-dimethyl-5-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)phenyl)-1H-pyrazole (22c)

[0153] 1,4-dimethyl-5-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazole(22c)

[0154]

[0155] Following conventional route C, using 5-(3-bromophenyl)-1,4-dimethyl-1H-pyrazole 20c (400 mg, 1.59 mmol) as the reactant, the target product 22c (412 mg, yield: 86.74%) was obtained.

[0156] Synthesis of 2-(2',6'-dimethyl-[1,1'-biphenyl]-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxoborane (22d)

[0157] 2-(2',6'-dimethyl-[1,1'-biphenyl]-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(22d)

[0158]

[0159] Following conventional route C, using 3'-bromo-2,6-dimethyl-1,1'-biphenyl 20d (400 mg, 2.30 mmol) as the reactant, the target product 22d (417 mg, yield: 88.3%) was obtained.

[0160] Synthesis of 8-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoborhecyclopentan-2-yl)phenyl)quinoline (22e)

[0161] 8-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)quinoline(22e)

[0162]

[0163] Following conventional route C, using 8-(3-bromophenyl)quinoline 20e (400 mg, 1.41 mmol) as the starting material, the target product 22e (372 mg, yield: 83.31%) was obtained.

[0164] Synthesis of 1-(cyclopropylmethyl)-3,5-dimethyl-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)phenyl)-1H-pyrazole (22g)

[0165] 1-(cyclopropylmethyl)-3,5-dimethyl-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-pyrazole(22g)

[0166]

[0167] Following conventional route C, using 20 g (400 mg, 1.31 mmol) of 4-(3-bromophenyl)-1-(cyclopropylmethyl)-3,5-dimethyl-1H-pyrazole as the reactant, 22 g (383 mg, yield: 83.0%) of the target product was obtained.

[0168] Example 15:

[0169] Synthesis of 4-(3-(1H-imidazo[4,5-b]pyridin-7-yl)phenyl)-3,5-dimethylisoxazole (Example 15)

[0170] 4-(3-(1H-imidazo[4,5-b]pyridin-7-yl)phenyl)-3,5-dimethylisoxazole (Example 15)

[0171]

[0172] Conventional synthetic route D: 7-Bromo-1H-imidazo[4,5-b]pyridine 23 (258 mg, 1.3 mmol), 3,5-dimethyl-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)phenyl)isoxazole 22a (506.73 mg, 1.69 mmol), cesium carbonate (848.66 mg, 2.60 mmol), and [1,1-bis(diphenylphosphine)ferrocene]palladium dichloromethane dichloride complex (106.38 mg, 0.13 mmol) were added to a Shrek tube, followed by 9 mL of dioxane and 3 mL of water as the reaction solvent. The air in the Shrek tube was completely replaced with argon, and the reaction was carried out at 100 °C for 7 h. The reaction was then monitored by TLC to indicate complete completion. After the reaction was cooled to room temperature, equal volumes of ethyl acetate and water were added for three extractions. The organic layers were combined and the solvent was removed by low-pressure concentration. The residue was subjected to column chromatography to obtain the target product Example 15 (197 mg, yield: 60.6%). 1 H NMR(300MHz,DMSO-d6)δ13.29(s,1H),8.53(s,1H),8.48–8.37(m,2H),8.29(d,J =7.9Hz,1H),7.72–7.61(m,2H),7.53(d,J=7.7Hz,1H),3.41(s,3H),2.35(s,3H).

[0173] Example 16:

[0174] Synthesis of 7-(3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 16)

[0175] 7-(3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 16)

[0176]

[0177] Following conventional route D, using 1,3,5-trimethyl-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)phenyl)-1H-pyrazole 22b (390 mg, 1.25 mmol) as the reactant, the target product Example 16 (232 mg, yield: 79.7%) was obtained. 1H NMR(300MHz,Chloroform-d)δ8.58(d,J=5.4Hz,1H),8.50(s,1H),8.10(d,J=7.9Hz,1H),8.02(s,1H),7 .65(t,J=7.7Hz,1H),7.56(d,J=5.1Hz,1H),7.42(d,J=7.7Hz,1H),3.86(s,3H),2.36(d,J=2.5Hz,6H).

[0178] Example 17:

[0179] Synthesis of 7-(3-(1,4-dimethyl-1H-pyrazol-5-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 17)

[0180] 7-(3-(1,4-dimethyl-1H-pyrazol-5-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 17)

[0181]

[0182] Following conventional route D, using 1,4-dimethyl-5-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)phenyl)-1H-pyrazole 22c (412 mg, 1.38 mmol) as the reactant, the target product Example 17 (192 mg, yield: 62.7%) was obtained. 1 HNMR(300MHz,Chloroform-d)δ8.59(d,J=5.2Hz,1H),8.43(s,1H),8.28(d,J=7.9Hz,1H),8.17(s,1H),7. 73(t,J=7.7Hz,1H),7.57(d,J=5.1Hz,1H),7.49(d,J=7.6Hz,1H),7.47(s,1H),3.92(s,3H),2.14(s,3H).

[0183] Example 18:

[0184] Synthesis of 7-(2',6'-dimethyl-[1,1'-biphenyl]-3-yl)-1H-imidazo[4,5-b]pyridine (Example 18)

[0185] 7-(2',6'-dimethyl-[1,1'-biphenyl]-3-yl)-1H-imidazo[4,5-b]pyridine (Example 18)

[0186]

[0187] Following conventional route D, using 2-(2',6'-dimethyl-[1,1'-biphenyl]-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxoborane 22d (417 mg, 1.35 mmol) as the reactant, the target product Example 18 (229.1 mg, yield: 69.2%) was obtained. 1 HNMR(300MHz,Chloroform-d)δ8.56(d,J=5.1Hz,1H),8.45(s,1H),8.38–8.31(m,1H),7.91(s,1H), 7.68(t,J=7.6Hz,1H),7.55(d,J=5.1Hz,1H),7.36–7.31(m,1H),7.21(q,J=5.8Hz,3H),2.17(s,6H).

[0188] Example 19:

[0189] Synthesis of 8-(3-(1H-imidazo[4,5-b]pyridin-7-yl)phenyl)quinoline (Example 19)

[0190] 8-(3-(1H-imidazo[4,5-b]pyridin-7-yl)phenyl)quinoline (Example 19)

[0191]

[0192] Following conventional route D, using 8-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)phenyl)quinoline 22e (372 mg, 1.17 mmol) as the reactant, the target product Example 19 (201.7 mg, yield: 69.6%) was obtained. 1 HNMR(300MHz,Chloroform-d)δ9.03(dd,J=4.2,1.8Hz,1H),8.49(d,J=5.2Hz,1H),8.37(t,J=1.7Hz,1H),8.31–8.23(m,2H),8. 10(d,J=7.7Hz,1H),7.87(ddd,J=14.3,7.6,1.5Hz,2H),7.79(dt,J=7.7,1.4Hz,1H),7.66(t,J=7.7Hz,2H),7.52–7.44(m,2H).

[0193] Example 20:

[0194] Synthesis of 7-(3-(1-(cyclopropylmethyl)-3,5-dimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 20)

[0195] 7-(3-(1-(cyclopropylmethyl)-3,5-dimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 20)

[0196]

[0197] Following conventional route D, using 22 g (387.68 mg, 1.10 mmol) of 1-(cyclopropylmethyl)-3,5-dimethyl-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)phenyl)-1H-pyrazole as the reactant, the target product Example 20 (149 mg, yield: 51.3%) was obtained. 1 H NMR (300MHz, DMSO-d6) δ13.24(s,1H),8.50(s,1H),8.44(d,J=5.1Hz,1H),8.30(s,1H),8.17(d,J=7.8Hz,1H),7.61(d,J=6.1Hz,2H) ,7.41(d,J=7.8Hz,1H),3.95(d,J=6.9Hz,2H),2.35(s,3H),2.25(s,3H),1.27(s,1H),0.55(d,J=7.6Hz,2H),0.41(d,J=4.9Hz,2H).

[0198] Synthesis Route 3:

[0199] Examples 21–24 were synthesized according to Scheme 3. Commercial starting materials 25a–25d and 19b were used as starting materials. The reaction was carried out according to step a of Scheme 2 synthetic route to obtain intermediates 26a–26d, borate ester intermediates 27a–27d, and the target compound. The synthesis of Examples 21–24 was carried out according to steps b and c of Scheme 2 synthetic route, respectively.

[0200]

[0201] Scheme 3.Reagents and conditions: (a) Cs2CO3, Pd(dppf)Cl2·CH2Cl2, 1,4-Dioxane, H2O, 100℃, 3h; (b) AcOK, Pd(dppf)Cl2·CH2Cl2, dry 1,4-Dioxane, 100℃, 3h; (c) Cs2CO3, Pd(dppf)Cl2·CH2Cl2, 1,4-Dioxane, H2O, 100℃, 7h.

[0202] Synthesis of 4-(3-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole (26a)

[0203] 4-(3-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole(26a)

[0204]

[0205] Following conventional route B, using 1-bromo-2-fluoro-3-iodobenzene 25a (530 mg, 1.76 mmol) and 1,3,5-trimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronan-2-yl)-1H-pyrazole 19b (540 mg, 2.29 mmol) as reactants, 4-(3-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole 26a (400 mg, 80.2%) was obtained.

[0206] Synthesis of 4-(5-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole (26b)

[0207] 4-(5-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole(26b)

[0208]

[0209] Following conventional route B, 4-bromo-1-fluoro-2-iodobenzene 25b (530 mg, 1.76 mmol) and 1,3,5-trimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronan-2-yl)-1H-pyrazole 19b (540 mg, 2.29 mmol) were used as reactants. The target product 26b (372 mg, 74.6%) was obtained.

[0210] Synthesis of 4-(3-bromo-2-chlorophenyl)-1,3,5-trimethyl-1H-pyrazole (26c)

[0211] 4-(3-bromo-2-chlorophenyl)-1,3,5-trimethyl-1H-pyrazole(26c)

[0212]

[0213] Following conventional route B, 1-bromo-2-chloro-3-iodobenzene 25c (530 mg, 1.67 mmol) and 1,3,5-trimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronan-2-yl)-1H-pyrazole 19b (512 mg, 2.17 mmol) were used as reactants. The target product 26c (381 mg, 76.2%) was obtained.

[0214] Synthesis of 4-(5-bromo-2,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrazole (26d)

[0215] 4-(5-bromo-2,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrazole(26d)

[0216]

[0217] Following conventional route B, 1-bromo-2,4-dichloro-5-iodobenzene 25d (530 mg, 1.66 mmol) and 1,3,5-trimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentan-2-yl)-1H-pyrazole 19b (510 mg, 2.16 mmol) were used as reactants. The target product 26d (391 mg, 78.1%) was obtained.

[0218] Synthesis of 4-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (27a)

[0219] 4-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(27a)

[0220]

[0221] Following conventional route C, using 4-(3-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole 26a (400 mg, 1.41 mmol) as the reactant, the target product 27a (366 mg, yield: 78.5%) was obtained.

[0222] Synthesis of 4-(2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (27b)

[0223] 4-(2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(27b)

[0224]

[0225] Following conventional route C, using 4-(5-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole 26b (400 mg, 1.41 mmol) as the reactant, the target product 27b (401 mg, yield: 85.9%) was obtained.

[0226] Synthesis of 4-(2-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (27c)

[0227] 4-(2-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(27c)

[0228]

[0229] Following conventional route C, using 4-(3-bromo-2-chlorophenyl)-1,3,5-trimethyl-1H-pyrazole 26c (400 mg, 1.34 mmol) as the reactant, the target product 27c (351 mg, yield: 75.8%) was obtained.

[0230] Synthesis of 4-(2,4-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (27d)

[0231] 4-(2,4-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(27d)

[0232]

[0233] Following conventional route C, using 4-(5-bromo-2,4-dimethylphenyl)-1,3,5-trimethyl-1H-pyrazole 26d (400 mg, 1.36 mmol) as the reactant, the target product 27d (376 mg, yield: 79.2%) was obtained.

[0234] Example 21:

[0235] Synthesis of 7-(2-fluoro-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 21)

[0236] 7-(2-fluoro-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 21)

[0237]

[0238] Following conventional route D, 4-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole 27a (279.5 mg, 0.85 mmol) was used as the reactant. The target product, Example 21 (138 mg, 66.0%), was obtained. 1 H NMR (300MHz, Chloroform-d) δ8.59(s,1H),8.40(s,1H),7.89(d,J=7.6Hz,1H),7.55(s,1H),7.38(p,J=7.5Hz,2H),3.84(s,3H),2.26(s,3H),2.23(s,3H).

[0239] Example 22:

[0240] Synthesis of 7-(4-fluoro-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 22)

[0241] 7-(4-fluoro-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 22)

[0242]

[0243] Following conventional route D, 4-(2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole 27b (279.5 mg, 0.85 mmol) was used as the reactant. The target product, Example 22 (113 mg, 54.1%), was obtained. 1 H NMR(300MHz,Chloroform-d)δ8.62(s,1H),8.40(s,1H),8.21(s,1H),8.06(d,J=6 .9Hz,1H),7.52(s,1H),7.38(d,J=9.0Hz,1H),3.85(s,3H),2.29(d,J=5.9Hz,6H).

[0244] Example 23:

[0245] Synthesis of 7-(2-chloro-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 23)

[0246] 7-(2-chloro-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 23)

[0247]

[0248] Following conventional route D, 4-(2-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole 27c (292.9 mg, 0.85 mmol) was used as the reactant. The target product, Example 23 (96 mg, 43.7%), was obtained. 1 H NMR (300MHz, Chloroform-d) δ8.59(s,1H),8.38(s,1H),7.61(d,J=7.5Hz,1H),7.48(s,2H),7.35(d,J=7.3Hz,1H),3.84(s,3H),2.21(s,3H),2.19(s,3H).

[0249] Example 24:

[0250] Synthesis of 7-(2,4-difluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 24)

[0251] 7-(2,4-difluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 24)

[0252]

[0253] Following conventional route D, 4-(2,4-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole 27d (294.2 mg, 0.85 mmol) was used as the reactant. The target product, Example 24 (105 mg, 47.6%), was obtained. 1 H NMR (300MHz, Chloroform-d) δ8.59(s,1H),8.38(s,1H),7.89(t,J=8.3Hz,1H),7.51(s,1H),7.12(t,J=9.9Hz,1H),3.83(s,3H),2.26(s,3H),2.25(s,3H).

[0254] Synthesis Route 4:

[0255] Examples 25–30 were synthesized according to Scheme 4. 28 was reacted with commercial starting materials 29a–29f in the presence of pyridine via a condensation reaction to yield intermediates 30a–30f. The synthesis of intermediates 31a–31f, borate ester intermediates 32a–32f, and the target product of Examples 25–30 were performed according to steps a, b, and c of the synthetic route in Scheme 2, respectively.

[0256]

[0257] Scheme 4.Reagents and conditions: (a) Pyridine, CH2Cl2, rt, 4h; (b) Cs2CO3, Pd(dppf)Cl2·CH2Cl2, 1,4-Dioxane, H2O, 100℃, 3h; (c) AcOK ,Pd(dppf)Cl2·CH2Cl2,dry1,4-Dioxane,100℃,3h; (d)Cs2CO3,Pd(dppf)Cl2·CH2Cl2,1,4-Dioxane,H2O,100℃,7h.

[0258] Synthesis of N-(3,5-dibromo-4-fluorophenyl)methanesulfonamide (30a)

[0259] N-(3,5-dibromo-4-fluorophenyl)methanesulfonamide(30a)

[0260]

[0261] Conventional Synthetic Route E: At room temperature, 2.0 g (7.44 mmol) of 3,5-dibromo-4-fluoroaniline 28 was placed in a 250 mL round-bottom flask, and 70 mL of dichloromethane was added as the reaction solvent, followed by pyridine (1.47 g / 1.49 mL, 18.6 mmol). Methylsulfonyl chloride 29a (937.1 mg, 8.18 mmol) was slowly added dropwise under constant stirring. After the addition was complete, the reaction was continued at room temperature for 4 h, and TLC was used to confirm the completeness of the reaction. The reaction mixture was extracted three times with 100 mL of 1 M dilute hydrochloric acid and an equal volume of ethyl acetate. The combined organic layers were extracted once with saturated sodium chloride, dried over anhydrous sodium sulfate for 1 h, and then concentrated under low pressure to remove the solvent. The residue was subjected to column chromatography to obtain the target product 30a (2.47 g, 95.7%).

[0262] Synthesis of N-(3,5-dibromo-4-fluorophenyl)ethanesulfonamide (30b)

[0263] N-(3,5-dibromo-4-fluorophenyl)ethanesulfonamide(30b)

[0264]

[0265] Following conventional route E, using ethylsulfonyl chloride 29b (1.04 g, 8.18 mmol) as the reactant, the target product 30b (2.56 g, yield: 95.3%) was obtained.

[0266] Synthesis of N-(3,5-dibromo-4-fluorophenyl)propane-1-sulfonamide (30c)

[0267] N-(3,5-dibromo-4-fluorophenyl)propane-1-sulfonamide(30c)

[0268]

[0269] Following conventional route E, using ethylsulfonyl chloride 29c (1.17 g, 8.18 mmol) as the reactant, the target product 30c (2.61 g, yield: 93.6%) was obtained.

[0270] Synthesis of N-(3,5-dibromo-4-fluorophenyl)cyclopropanesulfonamide (30d)

[0271] N-(3,5-dibromo-4-fluorophenyl)cyclopropanesulfonamide(30d)

[0272]

[0273] Following conventional route E, using cyclopropylsulfonyl chloride 29d (1.15 g, 8.18 mmol) as the starting material, the target product 30d (2.58 g, yield: 93.0%) was obtained.

[0274] Synthesis of N-(3,5-dibromo-4-fluorophenyl)-1-methyl-1H-pyrazole-4-sulfonamide (30e)

[0275] N-(3,5-dibromo-4-fluorophenyl)-1-methyl-1H-pyrazole-4-sulfonamide(30e)

[0276]

[0277] Following conventional route E, using 1-methyl-1H-pyrazole-4-sulfonyl chloride 29e (1.48 g, 8.18 mmol) as the starting material, the target product 30e (2.79 g, yield: 90.8%) was obtained.

[0278] Synthesis of N-(3,5-dibromo-4-fluorophenyl)pyridine-2-sulfonamide (30f)

[0279] N-(3,5-dibromo-4-fluorophenyl)pyridine-2-sulfonamide(30f)

[0280]

[0281] Following conventional route E, using 1-methyl-1H-pyrazole-4-sulfonyl chloride 29f (1.45 g, 8.18 mmol) as the starting material, the target product 30f (2.63 g, yield: 86.3%) was obtained.

[0282] Synthesis of N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)methanesulfonamide (31a)

[0283] N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)methanesulfonamide(31a)

[0284]

[0285] Following conventional route B, using N-(3,5-dibromo-4-fluorophenyl)methanesulfonamide 30a (600 mg, 1.73 mmol) as the reactant, the target product 31a (532 mg, yield: 81.8%) was obtained.

[0286] Synthesis of N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide (31b)

[0287] N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide(31b)

[0288]

[0289] Following conventional route B, using N-(3,5-dibromo-4-fluorophenyl)ethanesulfonamide 30b (600 mg, 1.66 mmol) as the reactant, the target product 31a (541 mg, yield: 83.4%) was obtained.

[0290] Synthesis of N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)propane-1-sulfonamide (31c)

[0291] N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)propane-1-sulfonamide(31c)

[0292]

[0293] Following conventional route B, using N-(3,5-dibromo-4-fluorophenyl)propane-1-sulfonamide 30c (600 mg, 1.60 mmol) as the reactant, the target product 31c (554 mg, yield: 85.7%) was obtained.

[0294] Synthesis of N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)cyclopropanesulfonamide (31d)

[0295] N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)cyclopropanesulfonamide(31d)

[0296]

[0297] Following conventional route B, using N-(3,5-dibromo-4-fluorophenyl)cyclopropanesulfonamide 30d (600 mg, 1.61 mmol) as the reactant, the target product 31d (547 mg, yield: 84.5%) was obtained.

[0298] Synthesis of N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazol-4-sulfonamide (31e)

[0299] N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazole-4-sulfonamide(31e)

[0300]

[0301] Following conventional route B, using N-(3,5-dibromo-4-fluorophenyl)cyclopropanesulfonamide 30e (600 mg, 1.45 mmol) as the reactant, the target product 31e (542 mg, yield: 84.4%) was obtained.

[0302] Synthesis of N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)pyridine-2-sulfonamide (31f)

[0303] N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)pyridine-2-sulfonamide(31f)

[0304]

[0305] Following conventional route B, using N-(3,5-dibromo-4-fluorophenyl)cyclopropanesulfonamide 30f (600 mg, 1.46 mmol) as the reactant, the target product 31f (517 mg, yield: 80.4%) was obtained.

[0306] Synthesis of N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)methylsulfonamide (32a)

[0307] N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)methanesulfonamide(32a)

[0308]

[0309] Following conventional route C, using N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)methanesulfonamide 31a (400 mg, 1.06 mmol) as the reactant, the target product 32a (351 mg, yield: 78.0%) was obtained.

[0310] Synthesis of N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide (32b)

[0311] N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide(32b)

[0312]

[0313] Following conventional route C, using N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide 31b (400 mg, 1.02 mmol) as the reactant, the target product 32b (347 mg, yield: 77.4%) was obtained.

[0314] Synthesis of N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)propane-1-sulfonamide (32c)

[0315] N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)propane-1-sulfonamide(32c)

[0316]

[0317] Following conventional route C, using N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)propane-1-sulfonamide 31c (400 mg, 0.98 mmol) as the reactant, the target product 32c (331 mg, yield: 74.1%) was obtained.

[0318] Synthesis of N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)cyclopropanesulfonamide (32d)

[0319] N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)cyclopropanesulfonamide(32d)

[0320]

[0321] Following conventional route C, using N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)cyclopropanesulfonamide 31d (400 mg, 0.99 mmol) as the reactant, the target product 32d (324 mg, yield: 72.5%) was obtained.

[0322] Synthesis of N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazol-4-sulfonamide (32e)

[0323] N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazole-4-sulfonamide(32e)

[0324]

[0325] Following conventional route C, using N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazol-4-sulfonamide 31e (400 mg, 0.91 mmol) as the reactant, the target product 32e (324 mg, yield: 73.2%) was obtained.

[0326] Synthesis of N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)pyridine-2-sulfonamide (32f)

[0327] N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)pyridine-2-sulfonamide(32f)

[0328]

[0329] Following conventional route C, using N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)pyridine-2-sulfonamide 31f (400 mg, 0.91 mmol) as the reactant, the target product 32f (342 mg, yield: 77.2%) was obtained.

[0330] Example 25:

[0331] Synthesis of N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)methanesulfonamide (Example 25)

[0332] N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)methanesulfonamide (Example 25)

[0333]

[0334] Following conventional route D, N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)methylsulfonamide 32a (300 mg, 0.71 mmol) was used as the reactant. The target product, Example 25 (193 mg, 65.7%), was obtained. 1 H NMR (300MHz, DMSO-d6) δ13.33(s,1H),9.96(d,J=9.5Hz,1H),8.52(s,1H),8.45(d,J=5.0Hz,1H),7.67(dd,J=5.9,2.8 Hz,1H),7.42(dd,J=5.0,1.8Hz,1H),7.22(dd,J=6.1,2.8Hz,1H),3.75(s,3H),3.12(s,3H),2.23(s,3H),2.14(s,3H).

[0335] Example 26:

[0336] Synthesis of N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide (Example 26)

[0337] N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide (Example 26)

[0338]

[0339] Following conventional route D, N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide 32b (300 mg, 0.68 mmol) was used as the reactant. The target product, Example 26 (203 mg, 69.1%), was obtained. 1HNMR(300MHz,DMSO-d6)δ13.32(s,1H),9.99(s,1H),8.57–8.41(m,2H),7.68(dd,J=5.9,2.8Hz,1H),7.43–7.32(m,1H), 7.22(dd,J=6.0,2.8Hz,1H),3.75(s,3H),3.21(dd,J=8.7,5.8Hz,2H),2.23(s,3H),2.13(s,3H),1.27(t,J=7.2Hz,3H).

[0340] Example 27:

[0341] Synthesis of N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)propane-1-sulfonamide (Example 27)

[0342] N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)propane-1-sulfonamide (Example 27)

[0343]

[0344] Following conventional route D, N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)cyclopropanesulfonamide 32c (300 mg, 0.66 mmol) was used as the reactant. The target product, Example 27 (164 mg, 55.8%), was obtained. 1 HNMR(300MHz,Chloroform-d)δ8.97(s,1H),8.54(d,J=5.1Hz,1H),8.38(s,1H),7.77(s,1H),7.48(d,J=4.5Hz,1H) ,3.78(s,3H),3.15(dd,J=9.4,6.3Hz,2H),2.16(s,3H),2.14(s,3H),1.90(h,J=7.4Hz,2H),1.03(d,J=7.4Hz,3H).

[0345] Example 28:

[0346] Synthesis of N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)cyclopropanesulfonamide (Example 28)

[0347] N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)cyclopropanesulfonamide (Example 28)

[0348]

[0349] Following conventional route D, N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)cyclopropanesulfonamide 32d (300 mg, 0.67 mmol) was used as the reactant. The target product, Example 28 (176 mg, 59.8%), was obtained. 1 H NMR(300MHz,DMSO-d6)δ9.92(s,1H),8.55(s,1H),8.47(d,J=5.0Hz,1H),7.63(s,1H),7.39(dd,J=5.0,1.4Hz,1H),7.25 (dd,J=6.2,2.8Hz,1H),3.75(s,3H),2.75(tt,J=7.2,5.2Hz,1H),2.23(s,3H),2.14(s,3H),1.00(dq,J=4.4,2.5Hz,4H).

[0350] Example 29:

[0351] Synthesis of N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazol-4-sulfonamide (Example 29)

[0352] N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazole-4-sulfonamide (Example 29)

[0353]

[0354] Following conventional route D, N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazol-4-sulfonamide 32e (300 mg, 0.61 mmol) was used as the reactant. The target product, Example 29 (182 mg, 61.8%), was obtained. 1 H NMR (300MHz, DMSO-d6) δ13.31(s,1H),10.59(s,1H),8.56–8.41(m,2H),7.91(dd,J=4.7,2.3Hz,1H),7.58(dd,J=5.9,2.8Hz,1H),7.33(dd, J=5.0,1.7Hz,1H),7.14(ddd,J=15.1,6.1,2.8Hz,1H),6.71(d,J=2.2Hz,1H),3.93(s,3H),3.73(s,3H),2.15(s,3H),2.05(d,J=2.9Hz,3H).

[0355] Example 30:

[0356] Synthesis of N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)pyridine-2-sulfonamide (Example 30)

[0357] N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)pyridine-2-sulfonamide (Example 30)

[0358]

[0359] Following conventional route D, N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)pyridine-2-sulfonamide 32f (300 mg, 0.62 mmol) was used as the reactant. The target product, Example 30 (158 mg, 53.6%), was obtained. 1H NMR (300MHz, DMSO-d6) δ13.35(s,1H),10.73(s,1H),9.09–8.78(m,2H),8.49(d,J=25.0Hz,2H),8.24( d,J=7.9Hz,1H),7.81–7.51(m,2H),7.33(s,1H),7.02(s,1H),3.72(s,3H),2.09(s,3H),1.99(s,3H).

[0360] Pharmacological activity evaluation:

[0361] 1. Inhibition rate test of BRD4(1) and BRD4(2)

[0362] The inhibitory activity of the compounds against BRD4(1) and BRD4(2) was evaluated by competitive fluorescence polarization (FP) assay. To establish FP detection, we designed and synthesized a FAM (MedChemExpress(MCE))-labeled fluorescent probe based on (+)-JQ1 (MedChemExpress(MCE)) (synthesis method is shown in Table 1 below). The example compounds (10 mM in DMSO (China National Pharmaceutical Group Chemical Reagent Co., Ltd.) stock solution) were prepared into solutions of various concentrations by serial dilution in assay buffer. BRD4(1) and BRD4(2) and the fluorescent probe (FAM-(+)-JQ1) were diluted to the desired concentrations in assay buffer. The experiment was performed in 384-well black flat-bottom polystyrene plates (Corning No. 3575). For each assay, equal volumes of diluted compound (20 μL), BRD4(1) (20 μL, 78 nM final) or BRD4(2) (20 μL, 53 nM final), and FAM-(+)-JQ1 (20 μL, 2.33 nM final) were sequentially added to the wells. The plate was capped and shaken at room temperature for 30 minutes. FP values ​​were measured using a SpectraMax multimode microplate reader (Molecular Devices) at excitation and emission wavelengths of 485 and 535 nm, respectively. For each assay, the FP value of the blank control (FAM-(+)-JQ1 only) was recorded as Pmin; the FP value of the negative control (FAM-(+)-JQ1 and protein) was recorded as Pmax; and the FP value of the test wells (compound, FAM-(+)-JQ1, and protein) was recorded as Ptest. The inhibition rate of the compound at each concentration point was calculated as follows: Inhibition rate (%) = [1 - (Ptest - Pmin) / (Pmax - Pmin)] × 100%. IC calculation was performed using Graphpad Prism 8.0 software. 50 .

[0363] Table 1. Inhibitory activity of the compounds in the examples against BRD4(1) and BRD4(2).

[0364]

[0365]

[0366] Synthesis method of fluorescent probe 5-FAM-(+)-JQ1:

[0367] Synthesis of (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diaza-6-yl)acetic acid (C1)

[0368] (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]di

[0369] azepin-6-yl)acetic acid(C1)

[0370]

[0371] (+)-JQ1 (300 mg, 656.47 μM) was dissolved in 10 mL of dichloromethane. 974 μL (20 eq) of trifluoroacetic acid was slowly added dropwise to the solution at room temperature, and the mixture was stirred for 3 h at room temperature. The reaction was then carried out by TLC until complete. The solvent was removed by concentration under reduced pressure to obtain a pale yellow oil (262 mg, 99.6%), which proceeded directly to the next step without further purification.

[0372] Synthesis of tert-butyl(S)-(2-(2-(2-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thiopheno[3,2-f][1,2)),4]triazolo[4,3-a][1,4]diaza-6-yl)acetamido)ethoxy)ethoxy)ethyl)carbamate (C3)

[0373] tert-butyl(S)-(2-(2-(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)ethoxy)ethoxy)ethyl)carbamate(C3)

[0374]

[0375] C1 (266 mg, 0.66 mmol) was dissolved in anhydrous dichloromethane at room temperature. pyBOP (379 mg, 0.73 mmol) and DIPEA (256 mg, 1.98 mmol) were added to the solution, and the mixture was stirred at room temperature for 30 minutes. Then, C2 (164 mg, 0.66 mmol) was added to the reaction mixture. After stirring for another 4 hours at room temperature, the reaction was monitored for completeness by TLC. The organic layer was extracted three times with water and dichloromethane. The combined organic layers were extracted with saturated NaCl, dried over anhydrous sodium sulfate for 30 minutes, and the solvent was removed by extraction and concentration. The resulting white powder, C3 (295 mg, 67.1%), was obtained by column chromatography. 1 H NMR (300MHz, DMSO-d6) δ7.51(d,J=8.8Hz,2H),7.45(d,J=8.6Hz,2H),6.81(t,J=5.7Hz,1H),4.54(dd,J=7.9,6.2Hz,1H),3.61–3.51(m,8H),3.48( t,J=5.8Hz,2H),3.41(t,J=6.1Hz,2H),3.10(t,J=5.9Hz,2H),3.06–2.98 (m,1H),2.62(s,3H),2.46–2.40(s,3H),1.68–1.62(s,3H),1.39(s,9H).

[0376] (s)-2-(2-(2-(2-(4-(4-chlorophenyl))-2,3,9-trimethyl-6H-thiophene[3,2-f][1,2,4]triazol[4,3-a][1,4]diazaphen-6-yl)acetamyl)ethoxy)ethoxy)ethane-1- Synthesis of -2,2,2-trifluoroacetate (C4)

[0377] (S)-2-(2-(2-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]dia zepin-6-yl)acetamido)ethoxy)ethoxy)ethan-1-aminium 2,2,2-trifluoroacetate(C4)

[0378]

[0379] At room temperature, C3 (295 mg, 0.44 mmol) was added to a 50 mL round-bottom flask, along with 10 mL of dichloromethane as the reaction solvent. 2 mL of trifluoroacetic acid was slowly added dropwise, and the reaction was monitored by TLC after stirring at 3 °C. The reaction was allowed to proceed to the next step at room temperature for 3 h. The solvent was removed under reduced pressure to obtain a brownish-red oil (210 mg). No further purification was required; the product was directly cast to the next step.

[0380] Synthesis of (S)-N-(2-(2-(2-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thiopheno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazaphen-6-yl)acetamido)ethoxy)ethoxy)ethyl)-3',6'-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9'-anthracene]-5-carboxamide (5-FAM-(+)-JQ1)

[0381] (S)-N-(2-(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)acet amido)ethoxy)ethoxy)ethyl)-3',6'-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-5-carboxamide(5-FAM-(+)-JQ1)

[0382]

[0383] At room temperature, C4 (210 mg, 0.37 mmol) was added to a 50 mL round-bottom flask, followed by 7 mL of anhydrous DMF, HATU (154.7 mg, 0.41 mmol), and DIPEA (144.5 mg, 1.1 mmol). The reaction was stirred for 30 minutes under N2 protection until a light brown mixture was formed. Then, 5-FAM (139.2 mg, 0.37 mmol) was added, and stirring continued for 6 hours. The reaction of the starting material was monitored by TLC. The organic layer was extracted three times with water and dichloromethane. The organic layers were combined and extracted with saturated NaCl. The organic layer was dried over anhydrous sodium sulfate for 30 min. The solvent was removed by extraction and concentration to obtain a pale yellow powder, 5-FAM-(+)-JQ1 (267 mg, 81.4%), which was separated by column chromatography. 1H NMR (300MHz, DMSO-d6) δ10.21(s,1H),8.95(t,J=5.4Hz,1H),8.49(s,1H),8.35(s,1H),8.28(dd, J=8.1,1.6Hz,1H),7.56–7.41(m,5H),7.39(d,J=8.1Hz,1H),6.72(d,J=2.2Hz,2H),6.61(d,J=8. 6Hz, 2H), 6.56 (dd, J=8.8, 2.2Hz, 2H), 4.53 (dd, J=7.8, 6.3Hz, 1H), 3.62–3.58 (m, 6H), 3.50 (q, J= 5.7,4.9Hz,4H),3.27(t,J=7.2Hz,4H),2.62(s,3H),2.43(s,3H),1.63(s,3H).Purity:97.95%by HPLC (MeOH / H2O=75:25). Purity: 97.93% by HPLC (MeOH / H2O=80:20, t R =3.535min).

[0384] 2. Cellular activity evaluation

[0385] Human myeloid monocytic leukemia cells MV-4-11, human ovarian cancer cells SKOV3, and human breast cancer cells MCF7 were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. MV-4-11 cells were cultured in IMDM (Adamas) medium, SKOV3 cells were cultured in McCoy's 5A medium (Gibco), and MCF7 cells were cultured in DMEM (Adamas) medium. All culture media were supplemented with 10% fetal bovine serum and 1% 100× penicillin-streptomycin antibiotics (15140-122, Gibco ThermoFisher, USA) and cultured at 37°C and 5% CO2. Cells in logarithmic growth phase were seeded at a density of 5000 cells per well into 96-well plates (Corning, model 3599) with a medium volume of 100 μL and incubated overnight at 37°C and 5% CO2. The following day, 100 μL of drug solutions prepared with different concentrations in culture medium were added, with three replicates for each concentration (denoted as RLU). test The study included control wells and blank wells. Control wells contained cells, culture medium, and a drug solution of the same concentration (denoted as RLU). control The blank wells contain culture medium (denoted as RLU). blankAfter 96 hours, 100 μL of culture medium was aspirated from each well and transferred to a 96-well plate. 100 μL of the CellTiter-Lumi luminescence assay kit (Beyotime Biotechnology) was then added to each well, and the plates were incubated on a shaker in the dark for 10 minutes before detection. For adherent cells, 100 μL of culture medium was aspirated from each well and discarded. 100 μL of the assay kit was added to each well, and the cells were lysed on a shaker for 2 minutes. The solution from each well was then aspirated to a 96-well plate, and the plates were incubated on a shaker in the dark for 8 minutes before detection. Chemiluminescence values ​​were detected using the chemiluminescence module on a Thermo Scientific Varioskan Flash. The percentage of viable cells was calculated using the following formula: Cell viability (%) = (RLU) / (RLU) test -RLU blank ) / (RLU control -RLU blank ()×100%, calculated using Graphpad Prism 8.0 software. 50 The results are shown in Table 2 and Figure 1 As shown.

[0386] Table 2 represents the in vitro antitumor activity of the MV4-11 cell line in the examples.

[0387]

[0388] The purpose of the above embodiments is to specifically illustrate the substantive content of the present invention, but those skilled in the art should know that the scope of protection of the present invention should not be limited to the specific embodiments.

Claims

1. A class of compounds with a biaromatic ring structure, which are compounds with the following chemical structures or pharmaceutically acceptable salts thereof:

2. The compound according to claim 1, characterized in that: The pharmaceutically acceptable salt is an acid addition salt of the compound with the chemical structure shown in claim 1, wherein the acid used for salt formation is selected from one of hydrogen chloride, sulfuric acid, hydrogen bromide, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid.

3. Use of the compound of claim 1 in the preparation of a BET inhibitor drug.

4. Use of the compound of claim 1 in the preparation of antitumor drugs or anti-inflammatory drugs.

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