A multi-substituted phenyl benzimidazo pyridine compound, its synthesis method and uses

Abstract

The present invention discloses a multi-substituted phenyl benzimidazo pyridine compound, a synthesis method and uses thereof. The multi-substituted phenyl benzimidazo pyridine compound provided by the present invention has a novel structure, has excellent inhibitory effects on BET proteins, and there is obvious selectivity in this inhibition, specifically, selectively inhibiting the BD1 domain of BET proteins. Those skilled in the art know that BET proteins are targets for tumor or inflammation treatment. Therefore, the multi-substituted phenyl benzimidazo pyridine compound provided by the present invention can be used to prepare anti-tumor drugs or anti-inflammatory drugs.

Description

Technical Field

[0001] The invention belongs to the field of medicinal chemistry, and specifically relates to a polysubstituted phenylbiimidazole pyridine compound and a synthesis method and use thereof. Background Art

[0002] The epigenetic target bromodomain and extra-terminal domain (BET) protein family is an extremely important target in the treatment of tumors and inflammation. The BET protein family contains four subtypes, namely BRD2, BRD3, BRD4 and BRDT, among which BRD4 is the most widely studied. BET family proteins all contain two highly conserved N-terminal tandem bromodomains BD1 and BD2. Each bromodomain consists of four antiparallel α helices (αZ, αA, αB and αC) and two hydrophobic loops (ZA loop and BC loop). Selective inhibition of one of the two bromodomains has become a focus of attention because it helps us understand the biological function of each bromodomain and thus rationally evaluate efficacy and toxicity.

[0003] Since BD1 and BD2 are two BDs connected in series on the same protein, it is difficult to examine the biological function of a single BD through conventional biological methods. Non-selective inhibitors of BD1 and BD2 have shown certain limitations in basic research and clinical trials. At present, there are more reports of BD2 selective inhibitors than BD1 selective inhibitors, and the activity and selectivity of reported BD1 selective inhibitors need to be further improved. The small number of BD1 selective inhibitors limits the understanding of the biological function and therapeutic value of selective inhibition of BD1, so the development of highly selective BD1 inhibitors and the exploration of their potential therapeutic effects are also urgent issues that need to be addressed. Summary of the invention

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a polysubstituted phenylbiimidazole pyridine compound and a synthesis method and use thereof.

[0005] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0006] A polysubstituted phenylbiimidazole pyridine compound is a compound represented by formula I or formula II, or a pharmaceutically acceptable salt of the compound represented by formula I or formula II:

[0007]

[0008] in:

[0009] Ring A is selected from phenyl or 5-10 membered aromatic hetero groups; a independently selected from hydrogen, C 1-3 Alkyl, n=1-5;

[0010] R 1, R 2 , R 3 are independently selected from hydrogen and halogen;

[0011] Y is independently selected from -CH 2 -、-(CH 2 ) m O-, -NR*-, -(CH 2 ) m S-; wherein m = 0, 1, 2, 3 or 4; R* is hydrogen, C 1-3 Alkyl or C 2-4 alkenyl;

[0012] R 4 are independently selected from hydrogen, 4-10 membered heterocycloalkane, R 3 - and R 4 The -Y- group forms a 5-10 membered cyclic ring with the benzene ring;

[0013]

[0014] in:

[0015] X is N or CH;

[0016] The C ring is independently selected from 5-6 membered aromatic heterocycles and 4-7 membered heterocycloalkanes; R c independently selected from hydrogen, halogen, C 1-3 Alkanes, C 1-3 Alkoxy, m=1-2;

[0017] R 5 independently selected from hydrogen, C 1-5 Alkanes, C 1-3 Alkoxy.

[0018] R 6 and R 7 Independently selected from hydrogen and halogen.

[0019] Preferably, the pharmaceutically acceptable salt is an acid addition salt of the compound of formula I or formula II, 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.

[0020] A method for synthesizing the above-mentioned polysubstituted phenylbiimidazole pyridine compound, the compound structure is shown in target 1, and the synthesis route is as follows:

[0021]

[0022] A method for synthesizing the above-mentioned multi-substituted phenylbiimidazole pyridine compound, the compound structure is shown in target 2 to target 3, and the synthesis route is as follows:

[0023]

[0024] A method for synthesizing the above-mentioned polysubstituted phenylbiimidazole pyridine compound, the compound structure is shown in target 4 to target 6, and the synthesis route is as follows:

[0025]

[0026] A method for synthesizing the above-mentioned multi-substituted phenylbiimidazole pyridine compound, the compound structure is shown in target 7 to target 8, and the synthesis route is as follows:

[0027]

[0028] A method for synthesizing the above-mentioned polysubstituted phenylbiimidazole pyridine compound, the compound structure is shown in target 9 to target 17, and the synthesis route is as follows:

[0029]

[0030] The polysubstituted phenylbiimidazole pyridine compounds are used for preparing BET inhibitor drugs.

[0031] Preferably, the inhibition is the selective inhibition of the BD1 domain of the BET protein.

[0032] The polysubstituted phenylbiimidazole pyridine compounds are used for preparing anti-tumor drugs or anti-inflammatory drugs.

[0033] Beneficial effects:

[0034] The polysubstituted phenyl biimidazole pyridine compounds provided by the present invention have novel structures, have excellent inhibitory effects on BET proteins, and this inhibition has obvious selectivity, specifically selectively inhibiting the BD1 domain of BET proteins. Those skilled in the art know that BET proteins are targets for tumor or inflammation treatment, and therefore, the polysubstituted phenyl biimidazole pyridine compounds provided by the present invention can be used to prepare anti-tumor drugs or anti-inflammatory drugs. DETAILED DESCRIPTION

[0035] The following examples are combined to specifically describe the essential contents of the present invention, but the protection scope of the present invention is not limited thereto. The experimental methods in the following examples without specifying specific conditions are implemented according to the existing methods and conditions in the industry.

[0036] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10 -6The unit of (ppm) is given. NMR measurements were performed using a Bruker AVANCE-300 nuclear magnetic spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD) as the measuring solvent, and tetramethylsilane (TMS) as the internal standard.

[0037] MS was measured using a liquid chromatography-mass spectrometer (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS). Aglient 6230 was used for HRMS.

[0038] The thin layer chromatography silica gel plate uses Qingdao GF254 silica gel plate, the silica gel plate used in thin layer chromatography (TLC) uses a specification of 0.15mm-0.2mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm-0.5mm. Other starting materials disclosed in the present invention can be synthesized according to methods known in the art, or from commercially available products.

[0039] Unless otherwise specified in the examples, the reactions can be carried out under argon or nitrogen atmosphere.

[0040] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0041] Synthetic route 1:

[0042] Example 1 was synthesized according to Scheme 1.

[0043]

[0044] Scheme 1.Reagents and conditions: (a)Cs 2 CO 3 ,Pd(dppf)Cl 2 ·CH 2 Cl 2 ,1,4-Dioxane,H 2 O, 100℃, 3h; (b) AcOK, Pd(dppf)Cl 2 ·CH 2 Cl 2 ,dry 1,4-Dioxane,100℃,3h; (c)Cs 2 CO 3 ,Pd(dppf)Cl 2 ·CH 2 Cl 2 ,1,4-Dioxane,H 2 O, 100℃, 7h.

[0045] 4-(7-Bromo-2,3-dihydrobenzofuran-5-yl)-3,5-dimethylisoxazole (34e)

[0046] 4-(7-bromo-2,3-dihydrobenzofuran-5-yl)-3,5-dimethylisoxazole(34e)

[0047]

[0048] Conventional synthetic route A: 33e (600 mg, 2.11 mmol), 19b (512.54 mg, 2.30 mmol), cesium carbonate (1.15 g, 3.53 mmol), and [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane 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 fully replaced with argon, and the reaction was completely completed after TLC monitoring at 100 ° C for 3 hours. After the reaction was cooled to room temperature, an equal volume of ethyl acetate and water were added for liquid extraction 3 times, the organic layers were combined and concentrated at low pressure to remove the solvent, and the residue was subjected to column chromatography to obtain the target product 34e (461 mg, yield: 84.9%).

[0049] 5-(3,5-Dimethylisoxazol-4-yl)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrofuran[2,3-c]pyridine (35e)

[0050] 5-(3,5-dimethylisoxazol-4-yl)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrofuro[2,3-c]pyridine(35e)

[0051]

[0052] Conventional synthetic route B: 34e (500 mg, 1.98 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) 21 (438.07 mg, 1.73 mmol), potassium acetate (260.47 mg, 2.65 mmol) and [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (108.79 mg, 0.13 mmol) were added to a Shrek tube, followed by 10 mL of anhydrous dioxane as a reaction solvent. The air in the Shrek tube was fully replaced with argon, and the reaction was completely completed after TLC monitoring at 100°C for 3 hours. After the reaction was cooled to room temperature, an equal volume of ethyl acetate and water were added for liquid extraction 3 times, the organic layers were combined and concentrated under low pressure to remove the solvent to obtain the residue 35e (466 mg, yield: 80.1%). Without further purification, it was directly used for the next step.

[0053] Embodiment 1:

[0054] 7-(5-(1,3,5-trimethyl-1H-pyrazol-4-yl)-2,3-dihydrobenzofuran-7-yl)-1H-imidazo[4,5-b]pyridine (Example 1)

[0055] 7-(5-(1,3,5-trimethyl-1H-pyrazol-4-yl)-2,3-dihydrobenzofuran-7-yl)-1H-imidazo[4,5-b]pyridine (Example 1)

[0056]

[0057] Conventional synthetic route C: 7-bromo-1H-imidazo[4,5-b]pyridine 23 (258 mg, 1.3 mmol), 35e (400 mg, 1.13 mmol), cesium carbonate (848.66 mg, 2.60 mmol) and [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane 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 reaction solvents. The air in the Shrek tube was fully replaced with argon, and the reaction was completely completed after TLC monitoring at 100°C for 7 hours. After the reaction was cooled to room temperature, an equal volume of ethyl acetate and water were added for liquid extraction 3 times, the organic layers were combined and concentrated at low pressure to remove the solvent, and the residue was subjected to column chromatography to obtain Example 1 (124 mg, yield: 41.5%). 1H NMR(300MHz,Chloroform-d)δ8.57(d,J=5.2Hz,1H),8.41(s,1H),7.74(s,1H),7.65(d,J=5.2Hz,1H),7.23 (s,1H),4.80(t,J=8.7Hz,2H),3.82(s,3H),3.41(t,J=8.7Hz,2H),2.32(d,J=1.7Hz,6H).HRMS(ESI):calcd for C 20 H 19 N 5 O[M+H] + 346.16, found 346.1672 Purity: 96.72% by HPLC (MeOH / H 2 O=65:35,t R =8.795min).

[0058] Synthesis route 2:

[0059] Examples 2-3 were synthesized according to Scheme 2.

[0060]

[0061] Scheme 3.Reagents and conditions: (a)Pd 2 (dba) 3 ,Xantphos,t-BuONa,toluene,100℃,5h(b)MeI,NaH,DMF,rt,overnight;(c)AcOK,Pd(dppf)Cl 2 ·CH 2 Cl 2 ,dry1,4-Dioxane,100℃,3h; (d)Cs 2 CO 3 ,Pd(dppf)Cl 2 ·CH 2 Cl 2 ,1,4-Dioxane,H 2 O, 100℃, 7h.

[0062] N-(3-Bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazol-4-amine (41)

[0063] N-(3-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazol-4-amine(41)

[0064]

[0065] At room temperature, 25a (500 mg, 1.97 mmol), 40 (271.16 mg, 2.17 mmol), Pd 2 (dba) 3 (90.1 mg, 0.09 mmol), Xantphos (113.8 mg, 0.197 mmol) and sodium tert-butoxide (378.63 mg, 3.94 mmol) were added to a Shrek tube, and 15 mL of toluene was added as the reaction solvent. After argon gas fully replaced the air in the reaction system, the reaction was carried out at 100° C. for 5 h. TLC detected that the reaction raw materials were completely reacted. An equal volume of water and ethyl acetate were added to extract the reaction solution three times. The organic layers were combined and extracted once with a saturated sodium chloride solution. The organic layer was dried over anhydrous sodium sulfate for 2 h. After suction filtration, the filtrate was concentrated under reduced pressure to remove the solvent to obtain a residue, which was chromatographed on silica gel to obtain the target product 41 (337 mg, yield: 57.4%).

[0066] N-(3-Bromo-2-fluorophenyl)-N,1,3,5-tetramethyl-1H-pyrazol-4-amine (42)

[0067] N-(3-bromo-2-fluorophenyl)-N,1,3,5-tetramethyl-1H-pyrazol-4-amine(42)

[0068]

[0069] At room temperature, 41 (300 mg, 1.01 mmol) was dissolved in DMF, and sodium hydride (120.7 mg, 3.02 mmol) was slowly added under stirring. After continuing stirring for 30 minutes, iodomethane (157.1 mg, 1.11 mmol) was added dropwise. After reacting at room temperature overnight, TLC detected that the reaction raw materials were completely reacted. Equal volumes of water and ethyl acetate were added to extract the reaction solution three times, the combined organic layers were extracted once with saturated sodium chloride solution, and the organic layer was dried over anhydrous sodium sulfate for 2 h. After suction filtration, the filtrate was concentrated under reduced pressure to remove the solvent to obtain the residue, which was chromatographed on silica gel to obtain the target product 42 (337 mg, yield: 57.4%).

[0070] N-(2-Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazol-4-amine (43a)

[0071] N-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazol-4-amine(43a)

[0072]

[0073] According to conventional route B, 41 (400.0 mg, 1.34 mmol) was used as the reaction raw material to obtain the target product 43a (312 mg, yield: 67.4%).

[0074] N-(2-Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-N,1,3,5-tetramethyl-1H-pyrazol-4-amine (43b)

[0075] N-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-N,1,3,5-tetramethyl-1H-pyrazol-4-amine(43b)

[0076]

[0077] According to conventional route B, 42 (400.0 mg, 1.28 mmol) was used as the reaction raw material to obtain the target product 43b (377 mg, yield: 81.9%).

[0078] Embodiment 2:

[0079] N-(2-Fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)phenyl)-1,3,5-trimethyl-1H-pyrazol-4-amine (Example 2)

[0080] N-(2-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)phenyl)-1,3,5-trimethyl-1H-pyrazol-4-amine (Example 2)

[0081]

[0082] According to the conventional route C, 43a (600 mg, 1.74 mmol) was used as the reaction raw material to obtain Example 2 (178 mg, yield: 39.6%). 1 H NMR (300 MHz, DMSO-d 6)δ13.25(s,1H),8.59–8.38(m,2H),7.44–7.29(m,1H),7.18–6.74(m,3H),6.48–6.32 (m,1H),3.73(s,3H),2.12(d,J=2.2Hz,3H),2.01(d,J=2.9Hz,3H).HRMS(ESI):calcd for C 18 H 17 FN 6 [M+Na] + 359.14, found 359.1392 Purity: 97.08% by HPLC (MeOH / H 2 O=80:20,t R =3.878min).

[0083] Embodiment 3:

[0084] N-(2-Fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)phenyl)-N,1,3,5-tetramethyl-1H-pyrazol-4-amine (Example 17)

[0085] N-(2-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)phenyl)-N,1,3,5-tetramethyl-1H-pyrazol-4-amine (Example 17)

[0086]

[0087] According to conventional route C, 43b (600 mg, 1.67 mmol) was used as the reaction raw material to obtain the target product Example 3 (219 mg, yield: 48.7%). 1 H NMR(300MHz,Chloroform-d)δ8.53(d,J=5.2Hz,1H),8.41(s,1H),7.45(d,J=4.9Hz,1H),7.28(s,1H),7.16 (t,J=7.9Hz,1H),6.93(t,J=8.4Hz,1H),3.74(s,3H),3.30(d,J=2.5Hz,3H),2.12(s,6H).HRMS(ESI):calcd for C 19 H 19 FN 6 [M+Na] + 373.15, found 373.1553 Purity: 94.37% by HPLC (MeOH / H 2 O=80:20,tR =6.250min).

[0088] Synthetic route 3:

[0089] Examples 4 to 6 were synthesized according to Scheme 3.

[0090]

[0091] Scheme 3.Reagents and conditions: (a) Isopropylmagnesium chloridelithium chloride complex (CAS:745038-86-2), dry THF, Ar, -40℃ to 0℃, overnight; (b) MeI, t-BuOK, DMF, 0℃ to rt, 6h; (c) AcOK, Pd(dppf)Cl 2 ·CH 2 Cl 2 ,dry 1,4-Dioxane,100℃,3h; (d)Cs 2 CO 3 ,Pd(dppf)Cl 2 ·CH 2 Cl 2 ,1,4-Dioxane,H 2 O, 100℃, 7h.

[0092] (3-Bromo-2-fluorophenyl)(oxetan-3-yl)methanol (45a)

[0093] (3-bromo-2-fluorophenyl)(oxetan-3-yl)methanol(45a)

[0094]

[0095] 44a (372.98 g, 4.33 mmol) was added into a 100 mL double-necked round-bottom flask, 40 mL of anhydrous tetrahydrofuran was used as solvent, the air in the reaction system was fully replaced with argon, the reaction system was cooled to -40 °C and maintained for 30 min, and a pre-cooled isopropylmagnesium chloride-lithium chloride (CAS: 745038-86-2) 2M tetrahydrofuran solution (5.1 mmol) was slowly added dropwise under a nitrogen flow, and the reaction was continued at -40 °C for 2 h. A tetrahydrofuran solution of a (1.0 g, 3.94 mmol) was slowly added dropwise to the above system, and the reaction was moved to an ice bath for 4 h. TLC showed that the reaction material was almost completely reacted. Saturated ammonium chloride was added to quench the reaction. Then, equal volumes of water and ethyl acetate were added to extract the reaction solution three times. The organic layers were combined and extracted once with saturated sodium chloride solution. The organic layer was dried over anhydrous sodium sulfate for 2 h. After filtration, the filtrate was concentrated under reduced pressure to remove the solvent to obtain a light yellow oil 45a, which was directly used for the next step without further purification.

[0096] (3-Bromo-2-fluorophenyl)(tetrahydrofuran-3-yl)methanol (45b)

[0097] (3-bromo-2-fluorophenyl)(tetrahydrofuran-3-yl)methanol(45b)

[0098]

[0099] According to the preparation method of 45a, 44b (433.75 mg, 4.33 mmol) was used as the reaction raw material to obtain the target product 45b as a light yellow oil.

[0100] (3-Bromo-2-fluorophenyl)(tetrahydro-2H-pyran-4-yl)methanol (45c)

[0101] (3-bromo-2-fluorophenyl)(tetrahydro-2H-pyran-4-yl)methanol(45c)

[0102]

[0103] According to the preparation method of 45a, 44c (494.5 mg, 4.33 mmol) was used as the reaction raw material to obtain the target product 45c as a light yellow oil.

[0104] 3-((3-Bromo-2-fluorophenyl)(methoxy)methyl)oxetane (46a)

[0105] 3-((3-bromo-2-fluorophenyl)(methoxy)methyl)oxetane(46a)

[0106]

[0107] At room temperature, 45a (300 mg, 1.15 mmol) was dissolved in 10 mL DMF, and potassium tert-butoxide was slowly added under stirring. After stirring at room temperature for 10 minutes, iodomethane (212.3 mg, 1.495 mmol) was added dropwise and the reaction was continued for 6 hours. TLC showed that the reaction material was almost completely reacted. Equal volumes of water and ethyl acetate were added to extract the reaction solution three times. The organic layers were combined and extracted once with saturated sodium chloride solution. The organic layer was dried over anhydrous sodium sulfate for 2 hours. After suction filtration, the filtrate was concentrated under reduced pressure to remove the solvent to obtain a residual oily substance, which was purified by silica gel column chromatography to obtain the target product 46a (310 mg, yield: 98.1%).

[0108] 3-((3-Bromo-2-fluorophenyl)(methoxy)methyl)tetrahydrofuran (46b)

[0109] 3-((3-bromo-2-fluorophenyl)(methoxy)methyl)tetrahydrofuran(46b)

[0110]

[0111] According to the preparation method of 46a, 45b (300 mg, 1.09 mmol) was used as the reaction raw material to obtain the target product 46b (302 mg, yield: 95.8%).

[0112] 4-((3-Bromo-2-fluorophenyl)(methoxy)methyl)tetrahydro-2H-pyran (46c)

[0113] 4-((3-bromo-2-fluorophenyl)(methoxy)methyl)tetrahydro-2H-pyran(46c)

[0114]

[0115] According to the preparation method of 46a, 45c (300 mg, 1.04 mmol) was used as the reaction raw material to obtain the target product 46c (303 mg, yield: 96.3%).

[0116] 2-(2-Fluoro-3-(methoxy(oxetan-3-yl)methyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (47a)

[0117] 2-(2-fluoro-3-(methoxy(oxetan-3-yl)methyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(47a)

[0118]

[0119] According to conventional route B, 46a (400 mg, 1.45 mmol) was used as the reaction raw material to obtain the target product 47a (413 mg, yield: 88.2%).

[0120] 2-(2-Fluoro-3-(methoxy(tetrahydrofuran-3-yl)methyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (47b)

[0121] 2-(2-fluoro-3-(methoxy(tetrahydrofuran-3-yl)methyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(47b)

[0122]

[0123] According to conventional route B, 46b (400 mg, 1.38 mmol) was used as the reaction raw material to obtain the target product 47a (401 mg, yield: 86.2%).

[0124] 2-(2-Fluoro-3-(methoxy(tetrahydro-2H-pyran-4-yl)methyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (47c)

[0125] 2-(2-fluoro-3-(methoxy(tetrahydro-2H-pyran-4-yl)methyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(47c)

[0126]

[0127] According to conventional route B, 46c (400 mg, 1.32 mmol) was used as the reaction raw material to obtain the target product 47c (409 mg, yield: 88.5%).

[0128] Embodiment 4:

[0129] 7-(2-Fluoro-3-(methoxy(oxetane-3-yl)methyl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 4)

[0130] 7-(2-fluoro-3-(methoxy(oxetan-3-yl)methyl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 4)

[0131]

[0132] According to the conventional route C, 47a (400 mg, 1.24 mmol) was used as the reaction raw material to obtain Example 4 (138 mg, yield: 46.1%). 1 H NMR (300 MHz, DMSO-d 6 )δ8.60–8.36(m,2H),7.85(t,J=7.3Hz,1H),7.62–7.48(m,2H),7.38(d,J=6.5Hz,1H),4.91(d,J=7.9Hz,1H),4.63(d, J=6.9Hz,2H),4.51(t,J=7.0Hz,1H),4.38(q,J=7.2,6.2Hz,1H),3.48–3.36(m,1H),3.27(s,3H).HRMS(ESI):calcdfor C 17 H 16 FN 3 O 2 [M+H] + 314.12, found 314.1283 Purity: 95.51% by HPLC (MeOH / H 2 O=80:20,t R =3.876min).

[0133] Embodiment 5:

[0134] 7-(2-Fluoro-3-(methoxy(tetrahydrofuran-3-yl)methyl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 5)

[0135] 7-(2-fluoro-3-(methoxy(tetrahydrofuran-3-yl)methyl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 5)

[0136]

[0137] According to the conventional route C, 47b (400 mg, 1.14 mmol) was used as the reaction raw material to obtain Example 5 (113 mg, yield: 37.8%). 1 H NMR(300MHz,Chloroform-d)δ8.58(dd,J=5.1,3.0Hz,1H),8.42(s,1H),7.95(tdd,J=6.9,4.6,1.7Hz,1H),7.61–7.50(m,2H),7.45–7.34(m,1H),4.58(dd ,J=8.1,4.3Hz,1H),4.01–3.58(m,4H),3.30(d,J=4.9Hz,3H),2.73(dp,J=11 .9,4.8,4.1Hz,1H),2.17–1.96(m,1H),1.88–1.60(m,1H).HRMS(ESI):calcd for C 18 H 18 FN 3 O 2 [M+H] + 328.14, found 328.1454 Purity: 98.62% by HPLC (MeOH / H 2 O=80:20,t R =3.874min).

[0138] Embodiment 6:

[0139] 7-(2-Fluoro-3-(methoxy(tetrahydro-2H-pyran-4-yl)methyl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 6)

[0140] 7-(2-fluoro-3-(methoxy(tetrahydro-2H-pyran-4-yl)methyl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 6)

[0141]

[0142] According to the conventional route C, 47c (400 mg, 1.13 mmol) was used as the reaction raw material to obtain Example 6 (132 mg, yield: 44.0%). 1H NMR(300MHz,Chloroform-d)δ8.56(d,J=5.1Hz,1H),8.41(d,J=8.4Hz,1H),7.93(td,J=7.3,1.9Hz,1H),7.57–7.48(m,2H),7.38(t,J=7.5Hz,1H),4.43( d,J=6.9Hz,1H),4.11–3.87(m,2H),3.48–3.31(m,2H),3.29(s,3H),1.90(d, J=12.6Hz,2H),1.58–1.51(m,2H),1.33(d,J=13.7Hz,1H).HRMS(ESI):calcd for C 19 H 20 FN 3 O 2 [M+H] + 342.16, found 342.1618 Purity: 95.47% by HPLC (MeOH / H 2 O=80:20,t R =3.524min).

[0143] Synthetic route 4:

[0144] Examples 7-8 were synthesized according to Scheme 4.

[0145]

[0146] Scheme 4.Reagents and conditions: (a)t-BuOK,DMF,rt,overnight; (b)AcOK,Pd(dppf)Cl 2 ·CH 2 Cl 2 ,dry 1,4-Dioxane,100℃,3h; (c)Cs 2 CO 3 ,Pd(dppf)Cl 2 ·CH 2 Cl 2 ,1,4-Dioxane,H 2 O, 100℃, 7h.

[0147] N-(3-Bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-N-methylethanesulfonamide (49a)

[0148] N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-N-methylethanesulfonamide(49a)

[0149]

[0150] At room temperature, N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide 31b (600 mg, 1.54 mmol) was dissolved in 20 mL DMF, and potassium tert-butoxide (345.03 mg, 3.07 mmol) was slowly added under stirring. After stirring for 30 min at room temperature, iodomethane (283.7 mg, 2.0 mmol) was added dropwise. After reacting at room temperature for 5 h, TLC detection showed that the reaction was completely completed. Equal volumes of ethyl acetate and water were added for three times of separation, the organic layers were combined, and the organic layers were separated and extracted again with saturated sodium chloride solution. The organic layer was taken, dried over anhydrous sodium sulfate for 1 h, and then concentrated to remove the solvent. After separation by column chromatography, a brown powder (587 mg, yield: 94.4%) was obtained.

[0151] N-(3-Bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide (49b)

[0152] N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide(49b)

[0153]

[0154] According to the preparation method of 49a, iodoethane (311.7 mg, 2.0 mmol) was used as the reaction raw material to obtain the target product 49b (569 mg, 88.47%).

[0155] N-(4-Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxolan-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-N-methylethylsulfonamide (50a)

[0156] 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)-N-methylethanesulfonamide(50a)

[0157]

[0158] According to conventional route B, N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)methanesulfonamide 49a (500 mg, 1.24 mmol) was used as the reaction raw material to obtain the target product 50a (421 mg, yield: 75.4%).

[0159] N-ethyl-N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxolan-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide (50b)

[0160] N-ethyl-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(50b)

[0161]

[0162] According to conventional route B, N-(3-bromo-4-fluoro-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide 49b (500 mg, 1.20 mmol) was used as the reaction raw material to obtain the target product 50b (432 mg, yield: 77.7%).

[0163] Embodiment 7:

[0164] N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-N-methylethanesulfonamide (Example 7)

[0165] N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-N-methylethanesulfonamide (Example 7)

[0166]

[0167] According to conventional route C, N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxolan-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-N-methylethylsulfonamide 50a (659.5 mg, 0.66 mmol) was used as the reaction raw material to obtain Example 7 (98 mg, yield: 43.85%). 1 H NMR (300MHz, Chloroform-d) δ8.57(d,J=17.3Hz,1H),8.39(s,1H),7.98(dd,J=5.9,2.8Hz,1H),7.55(d,J=5.2Hz,1H),7.40(dd,J=6. 0,2.9Hz,1H),3.84(s,3H),3.48(s,3H),3.20(q,J=7.4Hz,2H),2.28(dd,J=3.1,1.0Hz,6H),1.47(t,J=7.4Hz,3H).HRMS(ESI):calcd for C 21 H 23 FN 6 O 2 S[M+H] + 443.16, found 443.1665 Purity: 96.99% by HPLC (MeOH / H 2 O=80:20,t R =3.659min).

[0168] Embodiment 8:

[0169] N-ethyl-N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide (Example 8)

[0170] N-ethyl-N-(4-fluoro-3-(1H-imidazo[4,5-b]pyridin-7-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide (Example 8)

[0171]

[0172] According to conventional route C, N-ethyl-N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxolan-2-yl)-5-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)ethanesulfonamide 50b (305.5 mg, 0.66 mmol) was used as the reaction raw material to obtain Example 8 (103 mg, yield: 44.68%). 1 H NMR(300MHz,Chloroform-d)δ8.62(d,J=5.1Hz,1H),8.54(s,1H),7.95(s,1H),7.60(d,J=4.3Hz,1H),7.39(d,J=4.6Hz,1H),3.91 (d,J=7.0Hz,2H),3.87(s,3H),3.20(q,J=7.3Hz,2H),2.30(s,6H),1.49(t,J=7.2Hz,3H),1.31(t,J=7.0Hz,3H).HRMS(ESI):calcd for C 22 H 25 FN 6 O 2 S[M+H] + 457.17, found 457.1751 Purity: 94.08% by HPLC (MeOH / H 2 O=80:20,t R =4.408min).

[0173] Synthetic route 5:

[0174] Examples 9 to 17 were synthesized according to Scheme 5.

[0175]

[0176] Scheme 5.Reagents and conditions: (a)Cs 2 CO 3 ,Pd(dppf)Cl 2 ·CH 2 Cl 2 ,1,4-Dioxane,H 2 O, 100℃, 3h; (b) t-BuOK, DMSO, rt, overnight; (c) AcOK, Pd(dppf)Cl 2 ·CH 2 Cl 2 ,dry 1,4-Dioxane,100℃,3h; (d)Cs 2 CO 3, Pd(dppf)Cl 2 ·CH 2 Cl 2 , 1,4 - Dioxane, H 2 O, 100 °C, 7 h.

[0177] 4 - (3 - bromo - 2,5 - difluorophenyl)-1,3,5 - trimethylpyrazole(52)

[0178] 4 - (3 - bromo - 2,5 - difluorophenyl)-1,3,5 - trimethyl - 1H - pyrazole(52)

[0179]

[0180] According to the conventional route A, using 1,3 - dibromo - 2,5 - difluorobenzene 51 (500 mg, 1.84 mmol) and 19b (513.1 mg, 2.29 mmol) as reaction raw materials, the target product 52 (497 mg, yield: 89.8%) was obtained.

[0181] 4 - (3 - bromo - 2 - fluoro - 5 - (oxetan - 3 - yloxy)phenyl)-1,3,5 - trimethyl - 1H - pyrazole(54a)

[0182] 4 - (3 - bromo - 2 - fluoro - 5 - (oxetan - 3 - yloxy)phenyl)-1,3,5 - trimethyl - 1H - pyrazole(54a)

[0183]

[0184] Conventional reaction route D: At room temperature, 52 (450 mg, 1.49 mmol) and 53a (110 mg, 1.49 mmol) were added to a 100 mL round - bottom flask and 30 mL of DMSO was used as the reaction solvent. Potassium tert - butoxide (217.99 mg, 1.94 mmol) was slowly added under stirring and the reaction continued at room temperature overnight. After TLC detection, most of the reaction raw materials had reacted. Equal - volume saturated ammonium chloride solution and ethyl acetate were added for liquid - liquid extraction three times, and the organic layer was extracted with saturated sodium chloride solution once. The organic layer was dried over anhydrous sodium sulfate for 1 h, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was separated by silica gel column chromatography to obtain the target compound 54a (329 mg, yield: 62.0%).

[0185] 4 - (3 - bromo - 2 - fluoro - 5 - ((tetrahydrofuran - 3 - yl)oxy)phenyl)-1,3,5 - trimethyl - 1H - pyrazole(54b)

[0186] 4-(3-bromo-2-fluoro-5-((tetrahydrofuran-3-yl)oxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54b)

[0187]

[0188] According to the conventional route D, using 52 (450 mg, 1.49 mmol) and 19b (131.66 mg, 1.49 mmol) as reaction raw materials, the target product 52 (318 mg, yield: 57.6%) was obtained.

[0189] 4-(3-bromo-2-fluoro-5-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54c)

[0190] 4-(3-bromo-2-fluoro-5-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54c)

[0191]

[0192] According to the conventional route D, using 52 (450 mg, 1.49 mmol) and 53c (152.62 mg, 1.49 mmol) as reaction raw materials, the target product 54c (281 mg, yield: 49.1%) was obtained.

[0193] 4-(5-(azetidin-3-yloxy)-3-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole(54d)

[0194] 4-(5-(azetidin-3-yloxy)-3-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole(54d)

[0195]

[0196] According to the conventional route D, using 52 (450 mg, 1.49 mmol) and 53d (109.23 mg, 1.49 mmol) as reaction raw materials, the target product 54d (260 mg, yield: 49.1%) was obtained.

[0197] 4-(3-bromo-2-fluoro-5-((3-methyloxetan-3-yl)oxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54e)

[0198] 4-(3-bromo-2-fluoro-5-((3-methyloxetan-3-yl)oxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54e)

[0199]

[0200] According to the conventional route D, using 52 (450 mg, 1.49 mmol) and 53e (131.66 mg, 1.49 mmol) as reaction raw materials, the target product 54e (252 mg, yield: 45.7%) was obtained.

[0201] 4-(3-bromo-2-fluoro-5-(oxetan-3-ylmethoxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54f)

[0202] 4-(3-bromo-2-fluoro-5-(oxetan-3-ylmethoxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54f)

[0203]

[0204] According to the conventional route D, using 52 (450 mg, 1.49 mmol) and 53f (131.66 mg, 1.49 mmol) as reaction raw materials, the target product 54f (330 mg, yield: 59.8%) was obtained.

[0205] 4-(5-((2-oxaspiro[3.3]heptan-6-yl)oxy)-3-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole(54g)

[0206] 4-(5-((2-oxaspiro[3.3]heptan-6-yl)oxy)-3-bromo-2-fluorophenyl)-1,3,5-trimethyl-1H-pyrazole(54g)

[0207]

[0208] According to the conventional route D, using 52 (450 mg, 1.49 mmol) and 53g (170.57 mg, 1.49 mmol) as reaction raw materials, the target product 54g (279 mg, yield: 47.2%) was obtained.

[0209] 4-(3-bromo-2-fluoro-5-((3-methyloxetan-3-yl)methoxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54h)

[0210] 4-(3-bromo-2-fluoro-5-((3-methyloxetan-3-yl)methoxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54h)

[0211]

[0212] According to the conventional route D, using 52 (450 mg, 1.49 mmol) and 53h (152.62 mg, 1.49 mmol) as reaction raw materials, the target product 54h (306 mg, yield: 53.4%) was obtained.

[0213] 4-(3-bromo-2-fluoro-5-((tetrahydrofuran-2-yl)methoxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54i)

[0214] 4-(3-bromo-2-fluoro-5-((tetrahydrofuran-2-yl)methoxy)phenyl)-1,3,5-trimethyl-1H-pyrazole(54i)

[0215]

[0216] According to the conventional route D, using 52 (450 mg, 1.49 mmol) and 53i (152.62 mg, 1.49 mmol) as reaction raw materials, the target product 54i (361 mg, yield: 63.0%) was obtained.

[0217] 4-(2-fluoro-5-(oxetan-3-yloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(55a)

[0218] 4-(2-fluoro-5-(oxetan-3-yloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(55a)

[0219]

[0220] According to the conventional route B, using 54a (600 mg, 1.61 mmol) as the reaction raw material, the target product 55a (503 mg, yield: 74.0%) was obtained.

[0221] 4-(2-Fluoro-5-((tetrahydrofuran-3-yl)oxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (55b)

[0222] 4-(2-fluoro-5-((tetrahydrofuran-3-yl)oxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(55b)

[0223]

[0224] According to the conventional route B, using 54b (600 mg, 1.58 mmol) as the reaction raw material, the target product 55b (527 mg, yield: 77.9%) was obtained.

[0225] 4-(2-Fluoro-5-((tetrahydro-2H-pyran-4-yl)oxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (55c)

[0226] 4-(2-fluoro-5-((tetrahydro-2H-pyran-4-yl)oxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(55c)

[0227]

[0228] According to the conventional route B, using 54c (600 mg, 1.57 mmol) as the reaction raw material, the target product 55c (551 mg, yield: 81.8%) was obtained. 11H NMR (300 MHz, Chloroform-d) δ 8.56 (d, J = 5.1 Hz, 1H), 8.41 (s, 1H), 7.54 (s, 1H), 7.47 (s, 1H), 6.89 (s, 1H), 4.54 (s, 1H), 4.10–3.99 (m, 2H), 3.84 (s, 3H), 3.62 (t, J = 9.2 Hz, 2H), 2.28 (s, 3H), 2.26 (s, 3H), 2.11 (s, 2H), 1.96–1.82 (m, 2H).

[0229] 4-(5-(Azetidin-3-yloxy)-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (55d)

[0230] 4-(5-(azetidin-3-yloxy)-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (55d)

[0231]

[0232] Following the conventional route B, using 54d (600 mg, 1.63 mmol) as the reaction raw material, the target product 55d (522 mg, yield: 76.8%) was obtained.

[0233] 4-(2-Fluoro-5-((3-methyloxetan-3-yl)oxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (55e)

[0234] 4-(2-fluoro-5-((3-methyloxetan-3-yl)oxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (55e)

[0235]

[0236] Following the conventional route B, using 54e (600 mg, 1.59 mmol) as the reaction raw material, the target product 55e (551 mg, yield: 76.4%) was obtained.

[0237] 4-(5-(Cyclobutylmethoxy)-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (55f)

[0238] 4-(5-(cyclobutylmethoxy)-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(55f)

[0239]

[0240] According to the conventional route B, using 54f (600 mg, 1.63 mmol) as the reaction raw material, the target product 55f (551 mg, yield: 81.4%) was obtained.

[0241] 4-(5-((2-Oxaspiro[3.3]heptan-6-yl)oxy)-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (55g)

[0242] 4-(5-((2-oxaspiro[3.3]heptan-6-yl)oxy)-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(55g)

[0243]

[0244] According to the conventional route B, using 54g (600 mg, 1.49 mmol) as the reaction raw material, the target product 55g (534 mg, yield: 79.5%) was obtained.

[0245] 4-(2-Fluoro-5-((1-methylcyclobutyl)methoxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole (55h)

[0246] 4-(2-fluoro-5-((1-methylcyclobutyl)methoxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(55h)

[0247]

[0248] According to the conventional route B, using 54h (600 mg, 1.57 mmol) as the reaction raw material, the target product 55h (510 mg, yield: 75.7%) was obtained. 1 1H NMR (300 MHz, Chloroform-d) δ 8.53 (s, 1H), 8.43 (s, 1H), 7.49 (s, 1H), 7.43 (s, 1H), 6.88 (s, 1H), 4.67 (d, J = 5.9 Hz, 2H), 4.48 (d, J = 5.9 Hz, 2H), 4.11 (s, 2H), 3.81 (s, 3H), 2.24 (d, J = 6.0 Hz, 6H), 1.46 (s, 3H).

[0249] 4-(2-fluoro-5-((tetrahydrofuran-2-yl)methoxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(55i)

[0250] 4-(2-fluoro-5-((tetrahydrofuran-2-yl)methoxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1,3,5-trimethyl-1H-pyrazole(55i)

[0251]

[0252] According to the conventional route B, using 54i (600 mg, 1.57 mmol) as the reaction raw material, the target product 55i (503 mg, yield: 74.7%) was obtained.

[0253] Example 9:

[0254] 7-(2-fluoro-5-(oxetan-3-yloxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 9)

[0255] 7-(2-fluoro-5-(oxetan-3-yloxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 9)

[0256]

[0257] According to the conventional route C, using 55a (528.18 mg, 1.31 mmol) as the reaction raw material, Example 9 (182 mg, yield: 45.8%) was obtained. 1 H NMR (300 MHz, Chloroform-d) δ 8.65 (s, 1H), 8.48 (s, 1H), 7.59 (s, 1H), 7.36 (s, 1H), 6.71 (dd, J = 5.3, 2.8 Hz, 1H), 5.32 (m, 1H), 5.05 (t, J = 6.6 Hz, 2H), 4.89 (t, J = 6.1 Hz, 2H), 3.86 (s, 3H), 2.28 (d, J = 5.3 Hz, 6H). HRMS (ESI): calcd for C 21 H 20 FN 5 O 2 [M+H] + 394.16, found 394.1678 Purity: 95.90% by HPLC (MeOH / H 2 O = 80:20, t R = 4.876 min).

[0258] Example 10:

[0259] 7-(2-fluoro-5-((tetrahydrofuran-3-yl)oxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 10)

[0260] 7-(2-fluoro-5-((tetrahydrofuran-3-yl)oxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 10)

[0261]

[0262] According to the conventional route C, using 55b (546.88 mg, 1.31 mmol) as the reaction raw material, Example 10 (199 mg, yield: 48.4%) was obtained.1 1H NMR (300 MHz, Chloroform-d) δ 8.62 (d, J = 5.8 Hz, 2H), 7.65 (t, J = 4.8 Hz, 1H), 7.55 (d, J = 8.3 Hz, 1H), 7.11–7.03 (m, 1H), 3.99 (s, 1H), 3.87 (s, 3H), 3.50–3.33 (m, 2H), 3.24 (dd, J = 10.1, 4.4 Hz, 1H), 2.99 (p, J = 8.2 Hz, 1H), 2.31 (t, J = 3.2 Hz, 6H), 1.46–1.40 (m, 2H). HRMS (ESI): calcd for C 22 H 22 FN 5 O 2 [M+H] + 408.18, found 408.1825 Purity: 97.41% by HPLC (MeOH / H 2 O = 80:20, t R = 6.565 min).

[0263] Example 11:

[0264] 7-(2-Fluoro-5-((tetrahydro-2H-pyran-4-yl)oxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 11)

[0265] 7-(2-fluoro-5-((tetrahydro-2H-pyran-4-yl)oxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 11)

[0266]

[0267] Following the conventional route C, using 55c (565.01 mg, 1.31 mmol) as the reaction raw material, Example 11 (215 mg, yield: 50.5%) was obtained. 11H NMR (300 MHz, Chloroform-d) δ 8.56 (d, J = 5.1 Hz, 1H), 8.41 (s, 1H), 7.54 (s, 1H), 7.47 (s, 1H), 6.89 (s, 1H), 4.54 (s, 1H), 4.10–3.99 (m, 2H), 3.84 (s, 3H), 3.62 (t, J = 9.2 Hz, 2H), 2.28 (s, 3H), 2.26 (s, 3H), 2.11 (s, 2H), 1.96–1.82 (m, 2H). HRMS (ESI): calcd for C 23 H 24 FN 5 O 2 [M+H] + 422.20, found 422.2012 Purity: 94.88% by HPLC (MeOH / H 2 O = 80:20, t R = 4.805 min).

[0268] Example 12:

[0269] 7-(5-(Azetidin-3-yloxy)-2-fluoro-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 12)

[0270] 7-(5-(azetidin-3-yloxy)-2-fluoro-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 12)

[0271]

[0272] According to the conventional route C, using 55d (526.88 mg, 1.31 mmol) as the reaction raw material, Example 12 (201 mg, yield: 50.7%) was obtained. 1 1H NMR (300 MHz, DMSO-d 6)δ8.48(s,1H),8.42(d,J=5.0Hz,1H),7.38(dd,J=5.0,1.7Hz,1H),7.26(dd,J=5.5,3.1Hz,1H),6.77(dd,J=5.7,3.2Hz,1H),5.06(d,J=7.1Hz,1H),3.77–3.82(m,2H),3.73(s,3H),3.60(d,J=7.0Hz,2H),2.23–2.19(m,3H),2.12(s,3H).HRMS(ESI):calcd for C 21 H 21 FN 6 O[M+H] + 393.18,found 393.1832.Purity:99.25% by HPLC(MeOH / H 2 O=80:20,t R =4.082min).

[0273] Example 13:

[0274] 7-(2-fluoro-5-((3-methyloxetan-3-yl)oxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine(Example 13)

[0275] 7-(2-fluoro-5-((3-methyloxetan-3-yl)oxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine(Example 13)

[0276]

[0277] According to the conventional route C,using 55e(546.59mg,1.31mmol) as the reaction raw material,Example 13(191mg,yield:46.4%) was obtained. 1 H NMR(300MHz,Chloroform-d)δ8.68(d,J=43.7Hz,2H),8.47(s,1H),7.57(s,1H),6.61(s,1H),5.06(d,J=6.4Hz,2H),4.65(d,J=6.4Hz,2H),3.86(s,3H),2.28(d,J=5.7Hz,6H),1.88(s,3H).HRMS(ESI):calcd for C 22 H 22 FN5 O 2 [M+H] + 408.19, found 408.1942 Purity: 99.35% by HPLC (MeOH / H 2 O = 80:20, t R = 4.064 min).

[0278] Example 14:

[0279] 7-(2-Fluoro-5-(oxetan-3-ylmethoxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 14)

[0280] 7-(2-fluoro-5-(oxetan-3-ylmethoxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 14)

[0281]

[0282] According to the conventional route C, using 55f (546.59 mg, 1.31 mmol) as the reaction raw material, Example 14 (237 mg, yield: 57.6%) was obtained. 1 1H NMR (300 MHz, Chloroform-d) δ 8.65–8.45 (m, 1H), 8.36 (s, 1H), 7.54–7.44 (m, 2H), 6.85 (dd, J = 5.5, 3.1 Hz, 1H), 4.91 (dd, J = 7.8, 6.2 Hz, 2H), 4.60 (t, J = 6.1 Hz, 2H), 4.26 (d, J = 6.7 Hz, 2H), 3.80 (s, 3H), 3.47 (p, J = 6.9 Hz, 1H), 2.24 (s, 3H), 2.20 (s, 3H). 13 13C NMR (75 MHz, DMSO-d 6 ) δ 154.47, 148.87, 144.65, 144.35, 144.18, 137.69, 134.13, 132.90, 125.27, 125.04, 123.35, 118.36, 117.92, 116.82, 112.40, 73.65, 70.17, 36.38, 34.29, 12.79, 10.53. HRMS (ESI): calcd for C 22 1H 22 19F 5 1N2 [M+H] + 408.18, found 408.1837 Purity: 99.25% by HPLC (MeOH / H 2 O = 80:20, t R = 4.327 min).

[0283] Example 15:

[0284] 7-(5-((2-Oxaspiro[3.3]heptan-6-yl)oxy)-2-fluoro-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 15)

[0285] 7-(5-((2-oxaspiro[3.3]heptan-6-yl)oxy)-2-fluoro-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 15)

[0286]

[0287] According to the conventional route C, using 55 g (580.78 mg, 1.31 mmol) as the reaction raw material, Example 15 (182 mg, yield: 41.6%) was obtained. 1 H NMR (300 MHz, Chloroform-d) δ 8.55 (s, 1H), 8.33 (s, 1H), 7.53 (s, 1H), 7.39 (s, 1H), 6.71 (dd, J = 5.5, 3.2 Hz, 1H), 4.97–5.02 (m, 1H), 4.35 (dd, J = 9.7, 6.4 Hz, 2H), 4.10 (dd, J = 9.8, 4.2 Hz, 2H), 3.83 (s, 3H), 2.26 (s, 3H), 2.25–2.24 (m, 3H), 1.65–1.88 (m, 4H). 13 C NMR (75 MHz, DMSO-d 6 ) δ 152.24, 150.14, 148.89, 144.68, 144.38, 144.20, 137.78, 133.81, 132.86, 125.53, 118.34, 117.68, 116.89, 115.24, 112.24, 77.23, 70.87, 36.37, 31.47, 22.58, 14.47, 12.76, 10.51. HRMS (ESI): calcd for C 24 H 24FN 5 O 2 [M+H] + 434.20, found 434.1994 Purity: 92.86% by HPLC (MeOH / H 2 O = 80:20, t R = 6.250 min).

[0288] Example 16:

[0289] 7-(2-Fluoro-5-((3-methyloxetan-3-yl)methoxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 16)

[0290] 7-(2-fluoro-5-((3-methyloxetan-3-yl)methoxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 16)

[0291]

[0292] According to the conventional route C, using 55 h (565.32 mg, 1.31 mmol) as the reaction raw material, Example 16 (205 mg, yield: 48.2%) was obtained. 1 H NMR (300 MHz, Chloroform-d) δ 8.53 (s, 1H), 8.43 (s, 1H), 7.49 (s, 1H), 7.43 (s, 1H), 6.88 (s, 1H), 4.67 (d, J = 5.9 Hz, 2H), 4.48 (d, J = 5.9 Hz, 2H), 4.11 (s, 2H), 3.81 (s, 3H), 2.24 (d, J = 6.0 Hz, 6H), 1.46 (s, 3H). HRMS (ESI): calcd for C 23 H 24 FN 5 O 2 [M+H] + 422.20, found 422.1993 Purity: 98.90% by HPLC (MeOH / H 2 O = 80:20, t R = 3.606 min).

[0293] Example 17:

[0294] 7-(2-Fluoro-5-((tetrahydrofuran-2-yl)methoxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine (Example 17)

[0295] 7-(2-fluoro-5-((tetrahydrofuran-2-yl)methoxy)-3-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)-1H-imidazo[4,5-b]pyridine(Example 17)

[0296]

[0297] According to the conventional route C, using 55i (565.32 mg, 1.31 mmol) as the reaction raw material, Example 17 (221 mg, yield: 51.9%) was obtained. 1 H NMR (300 MHz, Chloroform-d) δ 8.58 (d, J = 5.0 Hz, 1H), 8.41 (s, 1H), 7.52 (d, J = 5.0 Hz, 1H), 7.45 (t, J = 4.3 Hz, 1H), 6.90 (dd, J = 5.7, 3.1 Hz, 1H), 4.35 (p, J = 6.1 Hz, 1H), 4.07 (d, J = 5.1 Hz, 2H), 3.99 (q, J = 7.1 Hz, 1H), 3.90 (t, J = 7.0 Hz, 1H), 3.83 (s, 3H), 2.25 (s, 3H), 2.22 (s, 3H), 2.13 (dt, J = 11.7, 6.7 Hz, 1H), 2.06–1.94 (m, 2H), 1.85 (s, 1H). HRMS (ESI): calcd for C 23 H 24 FN 5 O 2 [M+H] + 422.20, found 422.1989. Purity: 97.85% by HPLC (MeOH / H 2 O = 80:20, t R = 3.103 min).

[0298] Pharmacological activity evaluation :

[0299] 1. Testing the affinity of the compound of the example for BRD4(1) and BRD4(2) by MST method

[0300] Replace the protein buffer with the MonolithTM RED-NHS Second Generation Protein Labeling Kit (Nano Temper), and fluorescently label BRD4(1) and BRD4(2) proteins. Use a solution without primary amines (such as Tris or glycine), imidazole (Leyan), or β-mercaptoethanol (Aladdin) as the analysis buffer. Mix the fluorescently labeled protein with different concentrations of the compound in a 1:1 ratio. Set up a microscopic temperature gradient field with an infrared laser on the Monolith NT.115 (NanoTemper) analyzer, and track the movement of molecules such as fluorescent dye-labeled, tryptophan autofluorescence, and fluorescent fusion proteins in the microscopic temperature gradient field. Analyze the affinity of small molecules for BRD4(1) or BRD4(2) through the PR.ThermControl software.

[0301] Table 1 Binding Affinities of Examples for BRD4(1) and BRD4(2)

[0302]

[0303]

[0304] In Table 1, a Selectivty refers to the K of the compound for BRD4(2) D value / K of BRD4(1) D value.

[0305] 2. Activity Evaluation at the Cellular Level

[0306] Human myelomonocytic leukemia cells MV-4-11, human chronic myeloid leukemia cells K562, human ovarian cancer cells SKOV3, human colorectal cancer cells HCT-116, human pancreatic cancer cells MIAPaca-2, and human prostate cancer cells PC-3 were purchased from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences. Human non-small cell lung cancer cells A549 were purchased from Beyotime Biotechnology Company. MV-4-11 cells were cultured in IMDM medium, K562 and HCT-116 cells were cultured in RPMI-1640 medium, A549 and MIA Paca-2 cells were cultured in DMEM medium, SKOV3 cells were cultured in McCoy’s 5A medium, and PC-3 cells were cultured in Ham’s F-12 medium. When culturing, 10% fetal bovine serum and 1% of 100× penicillin-streptomycin double antibody (15140-122, Gibco ThermoFisher, USA) were added to each type of medium, and the cells were cultured at 37°C and 5% CO 2Cultured under the conditions. Cells in the logarithmic growth phase were seeded into 96-well plates at a density of 5000 cells per well, with a culture medium volume of 100 μL, and incubated overnight in an incubator at 37 °C and 5% CO 2 . The next day, different concentrations of drug solutions prepared with 100 μL of culture medium were added. Three replicate wells were set for each concentration (denoted as RLU test ), and control wells and blank wells were set. The control wells contained cells, culture medium, and the drug dissolution medium at the same concentration (denoted as RLU control ), and the blank wells contained only the culture medium (denoted as RLU blank ). After 96 h, for suspension cells, 100 μL was aspirated from each well into a 96-well black plate, and then 100 μL of the detection reagent of the CellTiter-Lumi luminescence cell viability detection kit (Beyotime Biotechnology) was added to each well. After incubation in the dark on a shaker for 10 min, the detection was carried out. For adherent cells, 100 μL of the culture medium was aspirated and discarded from each well. Then 100 μL of the detection reagent was added to each well in the 96-well plate. After lysis on a shaker for 2 min, the solution in the wells was aspirated into a 96-well black plate and incubated in the dark on a shaker for 8 min before detection. The chemiluminescence value was detected by the chemiluminescence module on the Thermoscientific varioskanflash. The percentage of cell survival was calculated using the following formula, and the IC 50 was calculated using Graphpad Prism 8.0 software.

[0307] Cell survival rate (%) = (RLU test - RLU blank ) / (RLU control - RLU blank ) × 100%

[0308] Table 2 Proliferation inhibitory activities of Example 14 against different tumor cells

[0309]

[0310] The role of the above examples is to specifically introduce the substantial content of the present invention. However, those skilled in the art should know that the protection scope of the present invention should not be limited to this specific example.

Claims

1. A multi-substituted phenyl benzimidazo[1,2-a]pyridine compound, characterized in that, it is a compound shown by the following chemical structure, or a pharmaceutically acceptable salt thereof:

2. The multi-substituted phenyl benzimidazo[1,2-a]pyridine compound according to claim 1, characterized in that: the pharmaceutically acceptable salt is an acid addition salt, and the acids used for salification include 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 multi-substituted phenyl benzimidazo[1,2-a]pyridine compound according to claim 1 for the preparation of a BET inhibitor drug.

4. According to the use described in claim 3, the inhibition is selective inhibition of the BD1 domain of the BET protein.

5. Use of the multi-substituted phenyl benzimidazo[1,2-a]pyridine compound according to claim 1 for the preparation of an anti-tumor drug or an anti-inflammatory drug.

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