CN113493407B

CN113493407B - A pyridine compound and its preparation method and pharmaceutical use

Info

Publication number: CN113493407B
Application number: CN202010258425.8A
Authority: CN
Inventors: 肖志艳; 景连栋; 张浩超; 杨亚军; 杨颖�
Original assignee: Institute of Materia Medica of CAMS and PUMC
Current assignee: Institute of Materia Medica of CAMS and PUMC
Priority date: 2020-04-03
Filing date: 2020-04-03
Publication date: 2025-01-28
Anticipated expiration: 2040-04-03
Also published as: CN113493407A

Abstract

The invention belongs to the technical field of medicines, and discloses a novel pyridine compound, a preparation method and a pharmaceutical application thereof. In particular to a novel pyridine compound shown in a formula (I), pharmaceutically acceptable salts thereof, a preparation method of the compound, application of the compound in preparation of CDK8 inhibitors, a pharmaceutical preparation containing the compound, and application of the compound in preparation of medicines for preventing and treating diseases related to tumors.

Description

Pyridine compound and preparation method and pharmaceutical application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to novel pyridine compounds shown in a general formula (I) and pharmaceutically acceptable salts thereof, pharmaceutical compositions and preparation methods of the compounds, and application of the compounds in preparation of cyclin-dependent kinase 8 (cyclin-DEPENDENT KINASE, CDK 8) inhibitors and in preparation of medicines for preventing or treating CDK8 related diseases.

Background

Malignant tumor has become one of serious diseases seriously threatening human health, and antitumor drugs play a great role in the clinical treatment process. However, as people's lifestyle, diet, environment, etc. change, the incidence and pathogenesis of malignant tumors also change significantly. Meanwhile, a certain kind of medicine is used for a long time, so that the tumor cells generate drug resistance. Therefore, the demand for antitumor drugs with novel structures and new action ways is increasing, and the development of such drugs is eager.

Cyclin dependent kinase 8 (CDK 8) is an important component of the mediator complex and plays a key role in the function of the mediator complex after binding to Cyclin C (Cyclin C) to form dimers. CDK8 is involved in multiple regulatory pathways, and is highly expressed in many cancer samples, and is considered as a potential target for the treatment of malignant tumors such as colorectal cancer, breast cancer, gastric cancer, ovarian cancer, and melanoma.

At present, small molecule inhibitors of CDK8 kinase have been reported, but drugs have not been successfully marketed due to the problems of unstable pharmacokinetic properties, poor selectivity and the like. In view of this, the inventors of the present application have sought to provide novel, safe and effective small molecule antitumor drugs with molecular targeting.

Disclosure of Invention

The technical problem solved by the invention is to provide a novel pyridine compound shown in a formula (I), a preparation method thereof, a pharmaceutical composition and application thereof in preparing a CDK8 inhibitor and application thereof in preparing a medicine for preventing or treating diseases related to hyperuricemia.

In order to solve the technical problems of the invention, the invention adopts the following technical scheme:

in a first aspect, the present invention provides a novel pyridine compound represented by the following general formula (I) and pharmaceutically acceptable salts thereof,

Wherein, Selected from double bond or single bond, R ₁ is selected from halogen, substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted indolyl, substituted or unsubstituted quinolinyl, substituted or unsubstituted indazolyl, the substituents are mono-or polysubstituted groups, each independently selected from halogen, -NH ₂、-OH、C_1-C₃ alkyl, C _1-C₃ alkoxy, morpholinyl, N-methylpiperazinyl, trifluoromethyl, cyano, nitro, R ₂ is selected from-NH ₂、-OH、-NHNH₂、C₁-C₃ alkoxy, C ₁-C₃ alkylamino, morpholinyl, N-methylpiperazinyl, - (CH ₂)_nCOOH,n＝1,2,3、-CHR₃COOH,R₃ is selected from hydrogen, C ₁-C₆ alkyl, C ₁-C₃ hydroxyalkyl.

Preferred compounds are those of the general formula (IA) and pharmaceutically acceptable salts thereof,

Wherein R ₁ is selected from halogen, substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted indolyl, substituted or unsubstituted quinolinyl, substituted or unsubstituted indazolyl, the substituents are mono-or polysubstituted groups, each independently selected from halogen, -NH ₂、-OH、C_1-C₃ alkyl, C _1-C₃ alkoxy, morpholinyl, N-methylpiperazinyl, trifluoromethyl, cyano, nitro, R ₂ is selected from-NH ₂、-OH、-NHNH₂、C₁-C₃ alkoxy, C ₁-C₃ alkylamino, morpholinyl, N-methylpiperazinyl, - (CH ₂)_nCOOH,n＝1,2,3、-CHR₃COOH,R₃ is selected from hydrogen, C ₁-C₆ alkyl, C ₁-C₃ hydroxyalkyl.

Preferred compounds are those of the general formula (IB) and pharmaceutically acceptable salts thereof,

More preferred compounds are the following compounds and pharmaceutically acceptable salts thereof, characterized in that said compounds are selected from the group consisting of:

According to a second aspect of the present invention, there is provided a method for synthesizing the compound of the first aspect, comprising the steps of:

the compound of the general formula (I) is obtained by taking 3-bromoisonicotinal as a starting material:

The definition of R ₁ and R ₂ is the same as that of the first aspect of the technical scheme.

According to a third aspect of the present invention, there is provided a pharmaceutical composition comprising an effective amount of any of the novel pyridines according to the first aspect of the present invention, and a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. The pharmaceutical composition is selected from tablets, capsules, pills, injections, sustained release preparations, controlled release preparations or various microparticle delivery systems.

For the preparation of medicaments, the compounds of the general formula (I) can be admixed in a known manner with suitable pharmaceutical carrier substances, fragrances, flavourings and colours in a known manner and formulated as tablets or coated tablets or suspended or dissolved in water or oil with other additional substances.

The compounds of the present invention may be administered orally or parenterally. The oral administration can be tablet, capsule, and coating agent, and parenteral administration dosage forms include injection and suppository. These formulations are prepared according to methods well known to those skilled in the art. Adjuvants used for the manufacture of tablets, capsules, and coatings are conventional adjuvants such as starch, gelatin, acacia, silica, polyethylene glycol, and solvents for liquid dosage forms such as water, ethanol, propylene glycol, and vegetable oils such as corn oil, peanut oil, olive oil, etc. Other adjuvants may also be present in the formulations containing the compounds of the invention, such as surfactants, lubricants, disintegrants, preservatives, flavouring agents, pigments and the like.

According to a fourth aspect of the present invention there is provided the use of a compound according to the first aspect of the invention, and a pharmaceutically acceptable salt thereof, in the preparation of a CDK8 inhibitor.

According to a fourth aspect of the present invention, there is also provided the use of the novel pyridines of the first aspect and pharmaceutically acceptable salts thereof in the manufacture of a medicament for the prophylaxis or treatment of a CDK8 related disorder. The CDK 8-related disease is selected from tumors and immune-related diseases. The tumor comprises rectal cancer, breast cancer, gastric cancer, ovarian cancer and melanoma.

Pharmacological studies indicate that the compounds of general formula (I) according to the invention have the activity of inhibiting CDK8, thus achieving therapeutic purposes.

The beneficial technical effects are as follows:

The CDK8 inhibitor has inhibiting effect on various tumor strains, and can be used for treating malignant tumors such as rectal cancer, breast cancer, gastric cancer, ovarian cancer, melanoma, etc. Poor specificity of action is one of the major challenges facing current research into CDK8 inhibitors. The compound has the characteristic and advantage of specifically inhibiting CDK8, and is expected to provide a novel safe and effective CDK8 inhibitor as a small-molecule antitumor drug.

Drawings

FIG. 1. Selective inhibition of CDK8 by a compound (single concentration inhibition of kinase by a compound of interest (10. Mu.M)).

Detailed Description

The invention is further illustrated below with reference to examples, which are not intended to limit the scope of the invention.

The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) or Mass Spectrometry (MS) or High Resolution Mass Spectrometry (HRMS). NMR displacements (δ) are given in parts per million (ppm). Column chromatography generally uses 200-300 mesh silica gel as a carrier. NMR measurements were performed using INOVA-300 with CDCl ₃、DMSO-d₆ and Acetone-d ₆ as measurement solvents, TMS as internal standard and chemical shifts given in ppm. MS was determined using AGILENT LC/MSD TOF LC-MS.

EXAMPLE 1 preparation of Compound 1

(1) Synthesis of ethyl (E) -3- (3-bromopyridin-4-yl) acrylate

Sodium hydride (content 60%,520 mg) was weighed and placed in a dry round bottom flask, 5mL of anhydrous tetrahydrofuran was added, and triethyl phosphono (2.912 g,13 mmol) dissolved in 20mL of anhydrous tetrahydrofuran was slowly added dropwise under ice bath, and the reaction was continued under stirring under ice bath for 1h. 3-bromo-4-aldehyde pyridine (1.86 g,10 mmol) diluted with 25mL tetrahydrofuran was continuously added dropwise to the reaction system, and the reaction was stirred at room temperature for about 3 hours after the completion of the dropwise addition. After the completion of the reaction, the solvent was distilled off, extracted with ethyl acetate and saturated brine, and the organic layer was combined and concentrated to give a pale yellow oil, which was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=20:1) to give a white solid (1.8 g, yield 70%).

(2) Synthesis of (E) -3- (3-bromopyridin-4-yl) acrylamide

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (77 mg,0.3 mmol) was dissolved in 2mL of 7M NH ₃ in methanol, and the reaction was stirred at room temperature for 24 hours, followed by addition of 2mL of NH ₃ in methanol. After the completion of the reaction, the solvent was distilled off, and the mixture was purified by silica gel column chromatography to give a white solid (38 mg, yield) 50％).¹H NMR(400MHz,DMSO-d₆)δ8.79(s,1H),8.56(d,J＝5.2Hz,1H),7.75(s,1H),7.64(d,J＝5.2Hz,1H),7.55(d,J＝15.6Hz,1H),7.40(s,1H),6.83(d,J＝15.6Hz,1H);¹³C NMR(100MHz,DMSO-d₆)δ165.31,152.25,148.87,141.79,134.55,129.45,121.62,121.62;HR-ESI-MS:m/z＝226.9824[M+H]⁺,calcd for C₈H₈BrN₂O:226.9815.

EXAMPLE 2 preparation of Compound 2

(1) Synthesis of ethyl (E) -3- (3- ((4- (4-methylpiperazin-1-yl) phenyl) amino) pyridin-4-yl) acrylate

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (512 mg,2 mmol), p-piperazinylaniline (420 mg,2.2 mmol), X-phos (95 mg,0.2 mmol), pd ₂(dba)₃ (92 mg,0.1 mmol), sodium t-butoxide (268 mg,2.8 mmol) were placed in a round bottom flask and 10mL toluene was added. The reaction system was purged with nitrogen for 5 minutes and then heated to reflux under nitrogen for about 8 hours. After the completion of the reaction, the reaction mixture was cooled to room temperature, the solvent was then distilled off, and the crude product was obtained by concentration of the organic layer by extraction with saturated brine and ethyl acetate, followed by silica gel column chromatography to give a solid (366 mg, yield 50%).

(2) Synthesis of (E) -3- (3- ((4- (4-methylpiperazin-1-yl) phenyl) amino) pyridin-4-yl) acrylamide

(E) Ethyl 3- (3- ((4- (4-methylpiperazin-1-yl) phenyl) amino) pyridin-4-yl) acrylate (110 mg,0.3 mmol) was dissolved in 2mL of 7M NH ₃ in methanol, and the reaction was stirred at room temperature for 24h, followed by addition of 2mL of NH ₃ in methanol. After the completion of the reaction, the solvent was distilled off, and the mixture was purified by silica gel column chromatography to give a white solid (45 mg, yield 45％).¹H NMR(400MHz,DMSO-d₆)δ8.45(s,1H),8.16(d,J＝5.2Hz,1H),7.94(s,1H),7.61(s,1H),7.55(d,J＝16.0Hz,1H),7.46(d,J＝5.2Hz,1H),7.21(s,1H),7.02(t,J＝8.0Hz,1H),6.71(d,J＝16.0Hz,1H),6.43-6.47(m,2H),6.27(dd,J＝8.0,1.2Hz,1H),3.05(t,J＝4.8Hz,4H),2.42(t,J＝4.8Hz,4H),2.20(s,3H);¹³C NMR(100MHz,DMSO-d₆)δ166.21,151.99,144.93,143.72,142.65,137.66,133.42,133.16,129.51,125.93,120.10,107.89,107.23,103.82,54.54,54.54,47.97,47.97,45.66;HR-ESI-MS:m/z＝338.1970[M+H]⁺,calcd for C₁₉H₂₄N₅O:338.1975.

EXAMPLE 3 preparation of Compound 3

(1) Synthesis of ethyl (E) -3- (3- ((4-fluorophenyl) amino) pyridin-4-yl) acrylate

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (512 mg,2 mmol), para-fluoroaniline (244 mg,2.2 mmol), X-phos (95 mg,0.2 mmol), pd ₂(dba)₃ (92 mg,0.1 mmol), sodium t-butoxide (268 mg,2.8 mmol) were placed in a round bottom flask and 10mL toluene was added. The reaction system was purged with nitrogen for 5 minutes and then heated to reflux under nitrogen for about 8 hours. After the completion of the reaction, the reaction mixture was cooled to room temperature, the solvent was then distilled off, and the crude product was obtained by concentration of the organic layer by extraction with saturated brine and ethyl acetate, followed by silica gel column chromatography to give a solid (300 mg, yield 50%).

(2) Synthesis of (E) -3- (3- ((4-fluorophenyl) amino) pyridin-4-yl) acrylamide

(E) Ethyl 3- (3- ((4-fluorophenyl) amino) pyridin-4-yl) acrylate (86 mg,0.3 mmol) was dissolved in 2mL of 7M NH ₃ in methanol, and the reaction was stirred at room temperature for 24h, followed by addition of 2mL of NH ₃ in methanol. After the completion of the reaction, the solvent was distilled off, and the mixture was purified by silica gel column chromatography to give a white solid (38 mg, yield) 45％).¹H NMR(400MHz,DMSO-d₆)δ8.40(s,1H),8.17(d,J＝4.8Hz,1H),8.05(s,1H),7.61(s,1H),7.56(d,J＝15.6Hz,1H),7.46(d,J＝5.2Hz,1H),7.21(s,1H),7.03-7.08(m,2H),6.90-6.96(m,2H),6.72(d,J＝15.6Hz,1H);¹³C NMR(100MHz,DMSO-d₆)δ166.17,142.85,142.65,140.55,140.54,137.91,133.03,132.95,126.12,120.18,118.44,118.37,115.87,115.65;HR-ESI-MS:m/z＝258.1036[M+H]⁺,calcd for C₁₄H₁₃FN₃O:258.1037.

EXAMPLE 4 preparation of Compound 4

(1) Synthesis of ethyl (E) -3- (3- (4-fluorophenyl) pyridin-4-yl) acrylate

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (1024 mg,4 mmol), and (4-fluorophenyl) boric acid (616mg,4.4mmol),Na₂CO₃(1272mg,12mmol),(PPh₃)₂PdCl₂(140mg,0.2mmol), were dissolved in a mixed solution of toluene, ethanol and water (volume ratio=1:1:0.2), and the mixed system was purged with nitrogen for 5 minutes. The reaction was then heated at reflux overnight under nitrogen. After the reaction, the solvent was distilled off, extracted with saturated brine and ethyl acetate, and the organic layer was combined and concentrated to give a solid crude product. The crude product was chromatographed on a column of silica gel (petroleum ether: ethyl acetate=10:1 elution) to give a white solid (564 mg, 52% yield).

(2) Synthesis of (E) -3- (3- (4-fluorophenyl) pyridin-4-yl) acrylic acid

(E) Ethyl 3- (3- (4-fluorophenyl) pyridin-4-yl) acrylate (678 mg,2.5 mmol) was dissolved in 10mL of ethanol, followed by addition of 7.5mL of 1N aqueous NaOH solution (7.5 mmol) to the system and stirring at room temperature for about 2h. After the reaction was completed, the organic solvent was distilled off, distilled water was added to the residue in an amount of about 5mL, the residue was completely dissolved, part of the impurities was removed by extraction with ethyl acetate, a water layer was separated, and a 1N aqueous HCl solution was added thereto to adjust the pH to weak acidity. After the completion of the adjustment, a large amount of white solid was precipitated, filtered and washed with distilled water, and dried to give a white solid product (528 mg, yield 87%).

(3) Synthesis of (E) -3- (3- (4-fluorophenyl) pyridin-4-yl) acryloyl-3-methoxyazetidine

(E) 3- (3- (4-fluorophenyl) pyridin-4-yl) acrylic acid (122 mg,0.5 mmol), 3-methoxyazetidine hydrochloride (52 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.5 mmol) in 5mL of dichloromethane and stirring at room temperature for about 6h. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a white solid product (94 mg, yield 60％).¹H NMR(400MHz,CDCl₃)δ8.60(d,J＝5.2Hz,1H),8.59(s,1H),7.56(d,J＝15.6Hz,1H),7.46(d,J＝5.2Hz 1H),7.29-7.32(m,1H),7.27-7.29(m,1H),7.14-7.20(m,2H),6.51(d,J＝16.0Hz,1H),4.36-4.40(m,1H),4.22-4.27(m,2H),4.09-4.11(m,1H),3.94-3.99(m,1H),3.33(s,3H);¹³C NMR(100MHz,DMSO-d₆)δ164.14,150.66,148.78,139.69,135.30,135.18,132.52,132.49,131.88,131.79,122.57,120.47,115.69,115.47,68.41,56.81,55.39,54.66;HR-ESI-MS:m/z＝313.1346[M+H]⁺,calcd for C₁₈H₁₈FN₂O₂:313.1347.

EXAMPLE 5 preparation of Compound 5

(E) Reaction of 3- (3- (4-fluorophenyl) pyridin-4-yl) acrylic acid with N-methylpiperazine gives the same synthesis and operation as compound 4 .¹H NMR(400MHz,CDCl₃)δ8.59(d,J＝5.2Hz,1H),8.57(s,1H),7.54(d,J＝15.6Hz,1H),7.46-7.48(m,1H),7.26-7.32(m,2H),7.13-7.18(m,2H),6.91(d,J＝15.6Hz,1H),3.70(s,2H),3.58(s,2H),2.41(t,J＝15.6Hz,4H),2.31(s,3H);¹³C NMR(100MHz,DMSO-d₆)δ163.48,150.67,148.70,139.88,136.65,135.14,132.61,132.58,131.89,131.81,123.47,120.44,115.67,115.45,55.02,54.25,45.54,44.92,41.57;HR-ESI-MS:m/z＝326.1658[M+H]⁺,calcd for C₁₉H₂₁FN₃O:326.1663.

EXAMPLE 6 preparation of Compound 6

(1) Synthesis of ethyl (E) -3- (3, 4-dimethoxyphenyl) pyridin-4-yl) acrylate

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (1024 mg,4 mmol), 3, 4-dimethoxyphenylboronic acid (800mg,4.4mmol),Na₂CO₃(1272mg,12mmol),(PPh₃)₂PdCl₂(140mg,0.2mmol), were dissolved in a mixed solution of toluene, ethanol and water (volume ratio=1:1:0.2), and the mixed system was purged with nitrogen for 5 minutes. The reaction was then heated at reflux overnight under nitrogen. After the reaction, the solvent was distilled off, extracted with saturated brine and ethyl acetate, and the organic layer was combined and concentrated to give a solid crude product. The crude product was chromatographed on a column of silica gel (petroleum ether: ethyl acetate=10:1 elution) to give a white solid (677 mg, 54% yield).

(2) Synthesis of (E) -3- (3, 4-dimethoxyphenyl) pyridin-4-yl) acrylic acid

(E) Ethyl 3- (3, 4-dimethoxyphenyl) pyridin-4-yl) acrylate (783 mg,2.5 mmol) was dissolved in 10mL of ethanol, followed by addition of 7.5mL of 1N aqueous NaOH (7.5 mmol) to the system and stirring at room temperature for about 2h. After the reaction was completed, the organic solvent was distilled off, distilled water was added to the residue in an amount of about 5mL, the residue was completely dissolved, part of the impurities was removed by extraction with ethyl acetate, a water layer was separated, and a 1N aqueous HCl solution was added thereto to adjust the pH to weak acidity. After the completion of the adjustment, a large amount of white solid was precipitated, filtered and washed with distilled water, and dried to give a white solid product (628 mg, yield 89%).

(3) Synthesis of (E) -3- (3, 4-dimethoxyphenyl) pyridin-4-yl) acryloyl-3-methoxyazetidine

(E) -3- (3, 4-dimethoxy phenyl) pyridine-4-yl) acrylic acid and 3-methoxy azetidine hydrochloride react, and the synthesis method and operation are the same as those of the compound 4 .¹H NMR(400MHz,CDCl₃)δ8.63(s,1H),8.57(d,J＝5.2Hz,1H),7.65(d,J＝15.6Hz,1H),7.46(d,J＝5.2Hz,1H),6.96(d,J＝8.0Hz,1H),6.83-6.87(m,2H),6.52(d,J＝15.6Hz,1H),4.37-4.41(m,1H),4.24-4.27(m,2H),4.10-4.13(m,1H),3.96-4.00(m,1H),3.93(s,3H),3.90(s,3H),3.33(s,3H);¹³C NMR(100MHz,DMSO-d₆)δ164.32,150.74,148.80,148.64,148.17,139.48,136.10,135.96,128.39,122.31,121.89,120.38,113.40,111.77,68.42,56.81,55.58,55.55,55.39,54.66;HR-ESI-MS:m/z＝355.1651[M+H]⁺,calcd for C₂₀H₂₃N₂O₄:355.1652.

EXAMPLE 7 preparation of Compound 7

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (1024 mg,4 mmol), 2-naphthalene boronic acid (757mg,4.4mmol),Na₂CO₃(1272mg,12mmol),(PPh₃)₂PdCl₂(140mg,0.2mmol), was dissolved in a mixed solution of toluene, ethanol and water (volume ratio=1:1:0.2), and the mixed system was purged with nitrogen for 5min. The reaction was then heated at reflux overnight under nitrogen. After the reaction, the solvent was distilled off, extracted with saturated brine and ethyl acetate, and the organic layer was combined and concentrated to give a solid crude product. The crude product was chromatographed on a silica gel column (petroleum ether: ethyl acetate=10:1 elution) to give a white solid (800 mg, yield 53％).¹H NMR(500MHz,DMSO-d₆)δ8.74(s,1H),8.68(d,J＝4.4Hz,1H),8.08(d,J＝6.8Hz,1H),8.05–8.00(m,2H),7.98(s,1H),7.94(d,J＝4.4Hz,1H),7.63–7.61(m,2H),7.52(d,J＝12.8Hz,1H),7.52(dd,J＝6.8,1.6Hz,1H),6.90(d,J＝12.8Hz,1H),4.14(q,J＝5.6Hz,2H),1.19(t,J＝5.6Hz,3H);¹³C NMR(125MHz,DMSO-d₆)δ165.62,151.05,149.00,139.95,138.87,136.43,133.48,132.81,132.36,128.99,128.21,128.20,127.71,127.67,126.91,126.88,123.42,120.49,60.49,14.10;HR-ESI-MS:m/z＝304.13345[M+H]⁺,calcd for C₂₀H₁₈O₂N:304.13375.

EXAMPLE 8 preparation of Compound 8

(E) Ethyl 3- (3- (naphthalen-2-yl) pyridin-4-yl) acrylate (758 mg,2.5 mmol) was dissolved in 5mL ethanol and 5mL tetrahydrofuran, followed by addition of 7.5mL 1N aqueous NaOH (7.5 mmol) to the system and stirring at room temperature for about 2h. After the reaction was completed, the organic solvent was distilled off, distilled water was added to the residue in an amount of about 5mL, the residue was completely dissolved, part of the impurities was removed by extraction with ethyl acetate, a water layer was separated, and a 1N aqueous HCl solution was added thereto to adjust the pH to weak acidity. After the completion of the adjustment, a large amount of white solid was precipitated, filtered and washed with distilled water, and dried to give a white solid (600 mg, yield) 87％).¹H NMR(400MHz,DMSO-d₆)δ12.70(s,1H),8.71(s,1H),8.67(d,J＝5.2Hz,1H),8.08(d,J＝8.4Hz,1H),8.00-8.04(m,2H),7.97(s,1H),7.92(d,J＝5.2Hz,1H),7.59-7.64(m,2H),7.52(dd,J＝8.4,1.6Hz,1H),7.46(d,J＝16.0Hz,1H),6.79(d,J＝15.6Hz,1H);¹³C NMR(100MHz,DMSO-d₆)δ166.82,150.92,148.91,139.29,139.06,136.29,133.52,132.74,132.27,128.83,128.11,128.11,127.62,127.58,126.78,126.77,124.53,120.36;HR-ESI-MS:m/z＝276.1019[M+H]⁺,calcd for C₁₈H₁₄O₂N:276.1019.

EXAMPLE 9 preparation of Compound 9

(E) The reaction of 3- (3- (naphthalene-2-yl) pyridin-4-yl) acrylic acid and methanol gives the same synthesis and operation as compound 4 .¹H NMR(400MHz,DMSO-d₆)δ8.71(s,1H),8.67(d,J＝5.2Hz,1H),8.06(d,J＝8.0Hz,1H),7.98-8.02(m,2H),7.95(s,1H),7.91(d,J＝5.2Hz,1H),7.59-7.61(m,2H),7.55-7.44(m,2H),6.87(d,J＝16.0Hz,1H),3.65(s,3H);¹³C NMR(100MHz,DMSO-d₆)δ165.96,150.98,148.93,139.95,138.80,136.35,133.42,132.74,132.29,128.86,128.14,128.12,127.62,127.56,126.82,126.79,123.10,120.44,51.75;HR-ESI-MS:m/z＝290.1173[M+H]⁺,calcd for C₁₉H₁₆NO₂:290.1176.

EXAMPLE 10 preparation of Compound 10

(E) The reaction of the methanol solution of the ethyl-3- (3- (naphthalene-2-yl) pyridine-4-yl) acrylate NH3 is carried out, and the synthesis method and the operation are the same as those of the compound 1 .¹H NMR(400MHz,Acetone-d₆)δ8.69(s,1H),8.63(d,J＝5.2Hz,1H),8.05(d,J＝8.4Hz,1H),7.98-8.03(m,2H),7.94(d,J＝1.6Hz,1H),7.69(d,J＝5.2Hz,1H),7.57-7.61(m,2H),7.55(d,J＝15.2Hz,1H),7.53(dd,J＝8.4,2.0Hz,1H),7.07(s,1H),6.91(d,J＝16.0Hz,1H),6.47(s,1H);¹³C NMR(100MHz,DMSO-d₆)δ166.16,151.40,149.41,140.44,136.68,135.37,134.22,133.25,132.75,129.18,128.59,128.54,128.09,128.09,127.93,127.19,127.19,120.35;HR-ESI-MS:m/z＝275.1178[M+H]⁺,calcd for C₁₈H₁₅N₂O:275.1179.

EXAMPLE 11 preparation of Compound 11

(E) Reaction of 3- (3- (naphthalene-2-yl) pyridin-4-yl) acrylic acid with methylamine hydrochloride, its synthesis method and operation are the same as compound 4 .¹H NMR(400MHz,CDCl₃)δ8.71(s,1H),8.63(d,J＝5.2Hz,1H),7.93(d,J＝8.8Hz,1H),7.87-7.92(m,2H),7.81(s,1H),7.59(s,1H),7.52-7.57(m,2H),7.49(d,J＝5.2Hz,1H),7.43(dd,J＝8.4,2.0Hz,1H),6.47(d,J＝15.6Hz,1H),5.62(s,1H),2.88(d,J＝4.8Hz,3H);¹³C NMR(100MHz,DMSO-d₆)δ164.41,150.09,148.08,140.94,136.41,133.92,133.44,132.73,132.31,128.75,128.11,128.09,127.68,127.61,127.54,126.79,126.74,120.23,25.66;HR-ESI-MS:m/z＝289.1334[M+H]⁺,calcd for C₁₉H₁₇N₂O:289.1335.

EXAMPLE 12 preparation of Compound 12

(E) The 3- (3- (naphthalene-2-yl) pyridin-4-yl) acrylic acid is obtained by reacting cyclopropylamine, and the synthesis method and operation are the same as those of the compound 4 .¹H NMR(400MHz,CDCl₃)δ8.71(s,1H),8.62(d,J＝5.3Hz,1H),7.95(s,1H),7.89(dt,J＝6.9,3.8Hz,2H),7.81(s,1H),7.60(d,J＝15.7Hz,1H),7.55(m,2H),7.50(d,J＝5.3Hz,1H),7.42(dd,J＝8.4,1.7Hz,1H),6.44(d,J＝15.7Hz,1H),5.74(s,1H),2.82-2.76(m,1H),0.80(dt,J＝6.9,5.5Hz,2H),0.56-0.51(m,2H);¹³C NMR(125MHz,DMSO-d₆)δ165.09,150.98,149.00,140.00,136.25,134.22,133.78,132.81,132.32,128.75,128.18,128.12,127.69,127.68,127.02,126.80,126.78,119.83,22.58,5.87,5.87;HR-ESI-MS:m/z＝315.15018[M+H]⁺,calcd for C₂₁H₁₉ON₂:315.14919.

EXAMPLE 13 preparation of Compound 13

(E) 3- (3- (naphthalen-2-yl) pyridin-4-yl) acrylic acid (138 mg,0.5 mmol), 3-aminoalanine methyl ester (84 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) were dissolved in 10mL of dichloromethane and the reaction stirred at room temperature for about 6h. After the completion of the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution and saturated brine, and the organic layer was concentrated to obtain a crude product, which was subjected to silica gel column chromatography to obtain a white solid product (110 mg, yield 61%). The white solid (110 mg,0.5 mmol) was dissolved in absolute ethanol, 2ml of 1M aqueous NaOH solution was added dropwise thereto, and the reaction was stirred at room temperature for about 2 hours. After the reaction, the reaction mixture was evaporated under reduced pressure, dissolved in distilled water, washed with ethyl acetate, and the aqueous phase was adjusted to pH 5 with 1M aqueous HCl to precipitate a white solid, which was suction-filtered to give a white solid (85 mg, yield 82％).¹H NMR(400MHz,DMSO-d₆)δ8.72(s,1H),8.70(d,J＝5.3Hz,1H),8.38(t,J＝5.6Hz,1H),8.07(d,J＝8.5Hz,1H),8.02(td,J＝6.9,4.4Hz,2H),7.96(s,1H),7.74(d,J＝5.3Hz,1H),7.65-7.59(m,2H),7.50(dd,J＝8.4,1.7Hz,1H),7.34(d,J＝15.7Hz,1H),6.91(d,J＝15.7Hz,1H),3.33(q,J＝6.6Hz,2H),2.42(t,J＝6.7Hz,2H);¹³C NMR(125MHz,DMSO-d₆)δ172.91,164.02,150.17,148.13,140.91,136.56,134.19,133.48,132.79,132.37,128.83,128.20,128.17,127.70,127.66,127.64,126.88,126.83,120.20,35.03,33.69;HR-ESI-MS:m/z＝347.13937[M+H]⁺,calcd for C₂₁H₁₉O₃N₂:347.13902.

EXAMPLE 14 preparation of Compound 14

(E) 3- (3- (naphthalen-2-yl) pyridin-4-yl) acrylic acid (138 mg,0.5 mmol), L-valine ethyl ester hydrochloride (109 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) in 10mL dichloromethane and stirred at room temperature for about 6h. After the reaction is finished, the reaction liquid is respectively washed by saturated sodium carbonate solution, 0.5N hydrochloric acid solution and saturated saline water, the organic layer is concentrated to obtain a crude product, and the crude product is subjected to silica gel column chromatography to obtain a light yellow oily substance. The pale yellow oil (116 mg,0.3 mmol) was dissolved in absolute ethanol, 2ml of 1M aqueous NaOH solution was added dropwise, and the reaction was stirred at room temperature for about 2h. After the reaction, the reaction mixture was evaporated under reduced pressure, dissolved in distilled water, washed with ethyl acetate, and the aqueous phase was adjusted to pH 5 with 1M aqueous HCl to precipitate a white solid, which was suction-filtered to give a white solid (110 mg, yield 73％).¹H NMR(400MHz,DMSO-d₆)δ8.70(s,1H),8.68(s,1H),8.38(d,J＝8.5Hz,1H),8.06(d,J＝8.6Hz,1H),8.01(ddd,J＝9.6,4.7,3.3Hz,2H),7.95(d,J＝1.7Hz,1H),7.68(d,J＝5.3Hz,1H),7.63-7.59(m,2H),7.50(dd,J＝8.4,1.8Hz,1H),7.35(d,J＝15.8Hz,1H),7.12(d,J＝15.7Hz,1H),4.21(dd,J＝8.5,5.6Hz,1H),2.13–2.04(m,1H),0.89(t,J＝6.9Hz,6H);¹³C NMR(100MHz,DMSO-d₆)δ172.66,164.12,150.89,148.87,139.74,136.21,134.67,133.61,132.65,132.17,128.61,128.00,127.97,127.52,127.52,126.71,126.63,126.63,119.51,57.25,29.81,19.01,17.85;HR-ESI-MS:m/z＝375.17050[M+H]⁺,calcd for C₂₃H₂₃O₃N₂:375.17032.

EXAMPLE 15 preparation of Compound 15

(E) Reaction of 3- (3- (naphthalene-2-yl) pyridin-4-yl) acrylic acid with dimethylamine hydrochloride, its synthesis method and operation are the same as compound 4 .¹H NMR(400MHz,Acetone-d₆)δ8.68(s,1H),8.62(d,J＝5.2Hz,1H),8.04(d,J＝8.4Hz,1H),7.97-8.02(m,2H),7.94(d,J＝1.6Hz,1H),7.83(d,J＝5.2Hz,1H),7.57-7.61(m,2H),7.55(d,J＝15.2Hz,1H),7.52(dd,J＝8.4,2.0Hz,1H),7.36(d,J＝15.4Hz,1H),3.18(s,3H),2.91(s,3H);¹³C NMR(100MHz,DMSO-d₆)δ164.66,150.82,148.70,140.02,136.38,136.33,133.78,132.69,132.19,128.69,128.04,128.00,127.59,127.56,126.70,126.66,123.72,120.44,36.82,35.28;HR-ESI-MS:m/z＝303.1491[M+H]⁺,calcd for C₂₀H₁₉N₂O:303.1492.

EXAMPLE 16 preparation of Compound 16

(E) Reaction of 3- (3- (naphthalene-2-yl) pyridin-4-yl) acrylic acid with 3-methoxyazetidine hydrochloride, its synthesis and operation are the same as those of compound 4 .¹H NMR(400MHz,CDCl₃)δ8.71(s,1H),8.64(d,J＝5.2Hz,1H),7.93(d,J＝8.4Hz,1H),7.86-7.92(m,2H),7.80(d,J＝1.2Hz,1H),7.59(d,J＝15.6Hz,1H),7.51-7.56(m,2H),7.50(d,J＝5.2Hz,1H),7.42(dd,J＝8.4,1.6Hz,1H),6.49(d,J＝15.6Hz,1H),4.26-4.30(m,1H),4.16-4.25(m,2H),3.99-4.02(m,1H),3.93(m,1H),3.28(s,3H);¹³C NMR(100MHz,DMSO-d₆)δ164.17,150.76,148.70,139.83,136.26,135.43,133.68,132.72,132.26,128.72,128.08,128.07,127.60,127.58,126.75,126.73,122.51,120.46,68.39,56.82,55.37,54.65;HR-ESI-MS:m/z＝345.1597[M+H]⁺,calcd for C₂₂H₂₁N₂O₂:345.1598.

EXAMPLE 17 preparation of Compound 17

(E) The reaction of 3- (3- (naphthalene-2-yl) pyridin-4-yl) acrylic acid and morpholine gives the same synthesis and operation as compound 4 .¹H NMR(400MHz,CDCl₃)δ8.71(s,1H),8.64(d,J＝5.2Hz,1H),7.94(d,J＝8.4Hz,1H),7.86-7.91(m,2H),7.81(d,J＝1.2Hz,1H),7.60(d,J＝15.6Hz,1H),7.53-7.57(m,2H),7.51(d,J＝5.2Hz,1H),7.43(dd,J＝8.4,2.0Hz,1H),6.81(d,J＝15.6Hz,1H),3.65(s,4H),3.57(s,2H),3.42(s,2H);¹³C NMR(100MHz,DMSO-d₆)δ163.65,150.85,148.72,139.93,136.99,136.2,133.80,132.73,132.23,128.71,128.08,128.05,127.64,127.60,126.71,126.71,123.11,120.41,66.30,66.01,45.57,42.05;HR-ESI-MS:m/z＝345.1598[M+H]⁺,calcd for C₂₂H₂₁N₂O₂:345.1598.

EXAMPLE 18 preparation of Compound 18

(E) Reaction of 3- (3- (naphthalen-2-yl) pyridin-4-yl) acrylic acid with N-methylpiperazine gives the same synthesis and operation as compound 4 .¹H NMR(400MHz,CDCl₃)δ8.71(s,1H),8.64(d,J＝5.2Hz,1H),7.94(d,J＝8.6Hz,1H),7.91-7.87(m,2H),7.81(s,1H),7.56-7.53(m,3H),7.52(d,J＝5.2Hz,1H),7.43(dd,J＝8.3,1.7Hz,1H),6.87(d,J＝15.6Hz,1H),3.67(s,2H),3.48(s,2H),2.35(d,J＝27.7Hz,4H),2.29(s,3H);¹³C NMR(125MHz,DMSO-d₆)δ163.39,150.75,148.62,139.84,136.74,136.10,133.72,132.63,132.13,128.61,127.99,127.96,127.56,127.51,126.62,126.62,123.26,120.34,54.91,54.13,45.44,44.80,41.44;HR-ESI-MS:m/z＝358.19046[M+H]⁺,calcd for C₂₃H₂₄ON₃:358.19139.

EXAMPLE 19 preparation of Compound 19

(E) Reaction of 3- (3- (naphthalene-2-yl) pyridin-4-yl) acrylic acid with L-proline methyl ester hydrochloride, its synthesis method and operation are the same as compound 4 .¹H NMR(400MHz,DMSO-d₆)δ8.68(d,J＝5.2Hz,2H),8.08-7.80(m,6H),7.60(dd,J＝7.2,2.4Hz,2H),7.49(d,J＝10.0Hz,1H),7.42-7.34(m,1H),7.29-7.04(m,1H),4.96-4.21(m,1H),3.75(m,2H),2.20-2.11(m,1H),2.00-1.76(m,3H);¹³C NMR(100MHz,DMSO-d₆)δ172.53,162.65,150.45,148.46,139.69,136.21,135.96,133.58,132.51,132.03,128.30,127.71,127.71,127.23,127.14,126.30,126.30,124.49,120.16,58.42,46.34,28.37,23.92;HR-ESI-MS:m/z＝373.15472[M+H]⁺,calcd for C₂₃H₂₁O₃N₂:373.15467.

EXAMPLE 20 preparation of Compound 20

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (1228 mg,4.8 mmol), 1-methyl-5-guide-azole-boronic acid (845mg,4.8mmol),Na₂CO₃(2035mg,19.2mmol),(PPh₃)₂PdCl₂(168mg,0.24mmol), was dissolved in a mixed solution of toluene, ethanol and water (volume ratio=1:1:0.2), and the mixed system was purged 5 times with nitrogen. Then the mixture is heated and refluxed for 12 hours in an oil bath under the protection of nitrogen. After the reaction, the solvent is distilled off, ethyl acetate is added, distilled water is used for washing, and the organic layer is combined and concentrated to obtain a solid crude product. The crude product was chromatographed on a silica gel column (petroleum ether: ethyl acetate=2:1 elution) to give a white solid (870 mg, yield 59％).¹H NMR(400MHz,DMSO-d₆)δ8.66(s,1H),8.63(d,J＝5.2Hz,1H),8.15(s,1H),7.89(d,J＝5.2Hz,1H),7.79(d,J＝8.7Hz,1H),7.78(d,J＝0.8Hz,1H),7.49(d,J＝16.0Hz,1H),7.39(dd,J＝8.6,1.6Hz,1H),6.86(d,J＝16.0Hz,1H),4.17-4.12(q,7.2Hz,2H),4.12(s,3H),1.20(t,J＝7.1Hz,3H).HR-ESI-MS:m/z＝308.13956[M+H]⁺,calcd for C₁₈H₁₈O₂N₃:308.13935.

EXAMPLE 21 preparation of Compound 21

(E) Ethyl-3- (3- (1-methyl-1H-indazol-5-yl) pyridin-4-yl) acrylate (616 mg,2.0 mmol) was dissolved in 20ml absolute ethanol, 3ml of 1M aqueous NaOH solution was added dropwise thereto, and the mixture was stirred at room temperature for 4 hours. After the completion of the reaction, the solvent was distilled off, dissolved in distilled water, washed with ethyl acetate, and the aqueous layer was adjusted to pH 5 with 1M aqueous HCl to give a pale yellow solid (470 mg, yield) 84％).¹H NMR(400MHz,DMSO-d₆)δ8.62(s,1H),8.61(d,J＝5.2Hz,1H),8.14(d,J＝0.9Hz,1H),7.84(d,J＝5.2Hz,1H),7.80-7.77(m,1H),7.77-7.76(m,1H),7.41-7.38(m,1H),7.37-7.35(m,1H),6.77-6.72(m,1H),4.11(s,3H);¹³C NMR(125MHz,DMSO-d₆)δ167.01,150.99,148.37,139.22,138.96,138.62,136.59,132.70,128.11,127.90,125.14,123.51,121.87,120.16,109.83,35.41;HR-ESI-MS:m/z＝280.10889[M+H]⁺,calcd for C₁₆H₁₄O₂N₃:280.10805.

EXAMPLE 22 preparation of Compound 22

(E) 3- (3- (1-methyl-1H-indazol-5-yl) pyridin-4-yl) acrylic acid (140 mg,0.5 mmol), methylamine hydrochloride (40.5 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) in 10mL dichloromethane and stirred at room temperature for about 6H. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (100 mg, yield 68％).¹H NMR(400MHz,DMSO-d₆)δ8.62(s,1H),8.61(d,J＝0.8Hz,1H),8.18,(d,J＝4.4Hz,1H),8.13(d,J＝0.9Hz,1H),7.78(dt,J＝8.6,0.9Hz,1H),7.75(dd,J＝1.6,0.8Hz,1H),7.64(d,J＝5.3Hz,1H),7.36(dd,J＝8.6,1.6Hz,1H),7.28(d,J＝15.7Hz,1H),6.78(d,J＝15.7Hz,1H),4.11(s,3H),2.66(d,J＝4.4Hz,3H);¹³C NMR(125MHz,DMSO-d₆)δ164.45,150.98,148.40,139.89,138.94,136.47,134.28,132.69,128.24,127.90,126.74,123.50,121.77,119.73,109.80,35.41,25.56;HR-ESI-MS:m/z＝293.13940[M+H]⁺,calcd for C₁₇H₁₇ON₄:293.13969.

EXAMPLE 23 preparation of Compound 23

(E) 3- (3- (1-methyl-1H-indazol-5-yl) pyridin-4-yl) acrylic acid (140 mg,0.5 mmol), morpholine (52 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) were dissolved in 10mL dichloromethane and the reaction stirred at room temperature for about 6H. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (110 mg, yield 63％).¹H NMR(400MHz,DMSO-d₆)δ8.64(s,1H),8.62(s,1H),8.14(d,J＝0.9Hz,1H),7.98(d,J＝5.3Hz,1H),7.78(dt,J＝8.7,0.9Hz,1H),7.76(dd,J＝1.6,0.8Hz,1H),7.47(d,J＝15.4Hz,1H),7.40-7.35(m,2H),4.11(s,3H),3.74-3.67(m,2H),3.59(dd,J＝8.1,4.0Hz,4H),3.52-3.46(m,2H);¹³C NMR(100MHz,DMSO-d₆)δ163.57,150.75,147.98,140.08,138.95,137.08,136.63,132.68,128.22,127.93,123.50,122.80,121.82,120.35,109.80,66.22,65.95,45.48,41.96,35.41;HR-ESI-MS:m/z＝349.16483[M+H]⁺,calcd for C₂₀H₂₁O₂N₄:349.16590.

EXAMPLE 24 preparation of Compound 24

(E) 3- (3- (1-methyl-1H-indazol-6-yl) pyridin-4-yl) acrylic acid (140 mg,0.5 mmol), methylamine hydrochloride (40.5 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) in 10mL dichloromethane and stirring at room temperature for about 6H. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (105 mg, yield 70％).¹H NMR(400MHz,DMSO-d₆)δ8.66-8.64(m,2H),8.65(d,J＝0.7Hz,1H),8.14(d,J＝0.9Hz,1H),7.86(dd,J＝8.3,0.8Hz,1H),7.71(q,J＝0.9Hz,1H),7.68-7.65(m,1H),7.29(d,J＝15.8Hz,1H),7.06(dd,J＝8.3,1.4Hz,1H),6.78(d,J＝15.8Hz,1H),4.08(s,3H),2.65(d,J＝4.7Hz,3H);¹³C NMR(125MHz,DMSO-d₆)δ164.40,150.84,148.80,139.93,139.51,136.47,134.06,134.01,132.33,126.93,122.77,122.53,120.68,119.62,110.54,35.34,25.56;HR-ESI-MS:m/z＝293.13937[M+H]⁺,calcd for C₁₇H₁₇ON₄:293.13969.

EXAMPLE 25 preparation of Compound 25

(E) 3- (3- (1-methyl-1H-indazol-6-yl) pyridin-4-yl) acrylic acid (140 mg,0.5 mmol), morpholine (52 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) were dissolved in 10mL dichloromethane and the reaction stirred at room temperature for about 6H. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (160 mg, yield 92％).¹H NMR(400MHz,DMSO-d₆)δ8.67-8.65(m,2H),8.14(d,J＝0.9Hz,1H),7.99(d,J＝5.3Hz,1H),7.87(dd,J＝8.3,0.8Hz,1H),7.71(d,J＝1.1Hz,1H),7.47(d,J＝15.4Hz,1H),7.37(d,J＝15.4Hz,1H),7.07(dd,J＝8.3,1.4Hz,1H),4.08(s,3H),3.72-3.66(m,2H),3.61-3.54(m,4H),3.52-3.46(m,2H);¹³C NMR(125MHz,DMSO-d₆)δ163.57,150.81,148.60,139.87,139.49,136.94,136.54,134.05,132.33,122.90,122.76,122.56,120.69,120.18,110.61,66.22,65.93,45.47,41.95,35.34;HR-ESI-MS:m/z＝349.16684[M+H]⁺,calcd for C₂₀H₂₁O₂N₄:349.16590.

EXAMPLE 26 preparation of Compound 26

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (1228 mg,4.8 mmol), indole-5-boronic acid (773mg,4.8mmol),Na₂CO₃(2035mg,19.2mmol),(PPh₃)₂PdCl₂(168mg,0.24mmol), was dissolved in a mixed solution of toluene, ethanol and water (volume ratio=1:1:0.2), and the mixed system was purged 5 times with nitrogen. Then the mixture is heated and refluxed for 12 hours in an oil bath under the protection of nitrogen. After the reaction, the solvent is distilled off, ethyl acetate is added, distilled water is used for washing, and the organic layer is combined and concentrated to obtain a solid crude product. The crude product was chromatographed on a silica gel column (petroleum ether: ethyl acetate=2:1 elution) to give a white solid (700 mg, yield 50％).¹H NMR(400MHz,DMSO-d₆)δ11.32(s,1H),8.64(s,1H),8.59(d,J＝5.2Hz,1H),7.86(d,J＝5.2Hz,1H),7.56(d,J＝16.0Hz,1H),7.54(s,1H),7.53(d,J＝4.4Hz,1H),7.46(t,J＝2.8Hz,1H),7.06(dd,J＝8.4,1.6Hz,1H),6.84(d,J＝16.0Hz,1H),6.53-6.51(m,1H),4.14(q,J＝7.2Hz,2H),1.20(t,J＝7.2Hz,3H).HR-ESI-MS:m/z＝293.12799[M+H]⁺,calcd for C₁₈H₁₇O₂N₂:293.12845.

EXAMPLE 27 preparation of Compound 27

(E) Ethyl 3- (3- (1H-indol-5-yl) pyridin-4-yl) acrylate (585 mg,2.0 mmol) was dissolved in 20ml absolute ethanol, 6ml of 1M aqueous NaOH was added dropwise thereto and the mixture was stirred at room temperature for 10 hours. After the reaction, the solvent was distilled off, dissolved in distilled water, washed with ethyl acetate, and the aqueous layer was adjusted to pH 5 with 1M aqueous HCl to give a pale yellow solid (225 mg, yield) 85％).¹H NMR(400MHz,DMSO-d₆)δ11.29(s,1H),8.45(s,1H),8.44(d,J＝5.2Hz,1H)7.81(d,J＝5.6Hz,1H),7.50(d,J＝5.6Hz,1H),7.48(s,1H),7.42(t,J＝2.4Hz,1H),7.05(d,J＝9.2Hz,1H),7.02(s,1H),6.73(d,J＝16.0Hz,1H),6.52(s,1H);¹³C NMR(125MHz,DMSO-d₆)δ169.69,151.51,148.21,142.53,137.88,136.29,135.98,131.38,128.34,128.13,126.87,123.59,121.80,120.24,111.98,101.98;HR-ESI-MS:m/z＝265.09689[M+H]⁺,calcd for C₁₆H₁₃O₂N₂:265.09715.

EXAMPLE 28 preparation of Compound 28

(E) 3- (3- (1H-indol-5-yl) pyridin-4-yl) acrylic acid (140 mg,0.5 mmol), methylamine hydrochloride (40.5 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) were dissolved in 10mL dichloromethane and the reaction stirred at room temperature for about 6H. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (80 mg, yield 58％).¹H NMR(400MHz,DMSO-d₆)δ11.29(s,1H),8.59(s,1H),8.57(d,J＝5.2Hz,1H),8.17(q,J＝4.8Hz,1H),7.61(d,J＝5.2Hz,1H),7.53-7.51(m,2H),7.45(t,J＝2.8Hz,1H),7.36(d,J＝15.6Hz,1H),7.04(dd,J＝8.4,1.6Hz,1H),6.77(d,J＝15.6Hz,1H),6.51-6.50(m,1H),2.66(d,J＝4.8Hz,3H);¹³C NMR(125MHz,DMSO-d₆)δ164.58,151.02,147.74,139.67,137.70,135.34,134.77,127.65,126.61,126.35,126.18,122.83,121.23,119.56,111.40,101.32,25.56;HR-ESI-MS:m/z＝278.12817[M+H]⁺,calcd for C₁₇H₁₆ON₃:278.12879.

EXAMPLE 29 preparation of Compound 29

(E) 3- (3- (1H-indol-5-yl) pyridin-4-yl) acrylic acid (132 mg,0.5 mmol), morpholine (52 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) dissolved in 10mL dichloromethane and stirred at room temperature for about 6H. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (100 mg, yield 60％).¹H NMR(400MHz,DMSO-d₆)δ11.32(s,1H),8.67(s,1H),8.64(d,J＝5.2Hz,1H),8.05(d,J＝5.2Hz,1H),7.54(s,1H),7.53-7.52(m,1H),7.48-7.45(m,3H),7.06(dd,J＝8.4,1.6Hz,1H),6.52-6.51(m,1H),3.71-3.70(m,2H),3.62-3.55(m,4H),3.50-3.49(m,2H);¹³C NMR(125MHz,DMSO-d₆)δ163.54,149.52,146.01,141.28,138.29,137.17,135.43,127.65,126.50,126.12,123.10,122.81,121.37,120.83,111.48,101.35,66.22,65.93,45.48,41.96;HR-ESI-MS:m/z＝334.15463[M+H]⁺,calcd for C₂₀H₂₀O₂N₃:334.15500.

EXAMPLE 30 preparation of Compound 30

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (1228 mg,4.8 mmol), indole-6-boronic acid pinacol ester (1215mg,5.0mmol),Na₂CO₃(2035mg,19.2mmol),(PPh₃)₂PdCl₂(168mg,0.24mmol), were dissolved in a mixed solution of toluene, ethanol and water (volume ratio=1:1:0.2), and the mixed system was purged 5 times with nitrogen. Then the mixture is heated and refluxed for 12 hours in an oil bath under the protection of nitrogen. After the reaction, the solvent is distilled off, ethyl acetate is added, distilled water is used for washing, and the organic layer is combined and concentrated to obtain a solid crude product. The crude product was chromatographed on a silica gel column (petroleum ether: ethyl acetate=2:1 elution) to give a white solid (1000 mg, yield 71％).¹H NMR(400MHz,DMSO-d₆)δ11.31(s,1H),8.66(s,1H),8.60(d,J＝5.2Hz,1H),7.87(d,J＝5.2Hz,1H),7.68(d,J＝8.0Hz,1H),7.57(d,J＝16.0Hz,1H),7.46(t,J＝2.4Hz,1H),7.38(s,1H),6.98(dd,J＝8.0,1.6Hz,1H),6.85(d,J＝16.0Hz,1H),6.53(t,J＝2.4Hz,1H),4.14(q,J＝7.2Hz,2H),1.20(t,J＝7.2Hz,3H);HR-ESI-MS:m/z＝293.12872[M+H]⁺,calcd for C₁₈H₁₇O₂N₂:293.12845.

EXAMPLE 31 preparation of Compound 31

(E) Ethyl 3- (3- (1H-indol-6-yl) pyridin-4-yl) acrylate (585 mg,2.0 mmol) is dissolved in 20ml absolute ethanol, 4ml of 1M aqueous NaOH solution is added dropwise and the mixture is stirred at room temperature for 6H. After the completion of the reaction, the solvent was distilled off, dissolved in distilled water, washed with ethyl acetate, and the aqueous layer was adjusted to pH 5 with 1M aqueous HCl to give a pale yellow solid (423 mg, yield) 80％).¹H NMR(400MHz,DMSO-d₆)δ11.31(s,1H),8.47(s,1H),8.46(d,J＝5.2Hz,1H),7.64(d,J＝1.6Hz,1H),7.62(d,J＝5.2Hz,1H),7.41(t,J＝2.4Hz,1H),7.33(s,1H),7.07,(dd,J＝15.6,1.2Hz,1H),6.95(dd,J＝8.0,1.6Hz,1H),6.56(dd,J＝15.6,1.2Hz,1H),6.49(t,J＝2.4Hz,1H);¹³C NMR(125MHz,DMSO-d₆)δ169.07,150.65,147.58,141.71,136.89,135.69,135.47,130.71,129.35,126.98,126.18,120.73,119.80,119.55,112.54,100.83;HR-ESI-MS:m/z＝265.09796[M+H]+,calcd for C₁₆H₁₃O₂N₂:265.09715.

EXAMPLE 32 preparation of Compound 32

(E) 3- (3- (1H-indol-6-yl) pyridin-4-yl) acrylic acid (140 mg,0.5 mmol), methylamine hydrochloride (40.5 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) were dissolved in 10mL dichloromethane and the reaction stirred at room temperature for about 6H. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (78 mg, yield 57％).¹H NMR(400MHz,DMSO-d₆)δ11.28(s,1H),8.61(s,1H),8.58(d,J＝5.2Hz,1H),8.18(q,J＝4.4Hz,1H),7.66(d,J＝8.4Hz,1H),7.62(d,J＝5.2Hz,1H),7.45(m,1H),7.38(d,J＝15.6Hz,1H),7.36-7.35(m,1H),6.97(dd,J＝8.4,1.6Hz,1H),6.78(d,J＝15.6Hz,1H),6.52-6.51(m,1H),2.67(d,J＝4.4Hz,3H);¹³C NMR(125MHz,DMSO-d₆)δ164.56,150.97,147.94,139.64,137.42,135.67,134.81,128.60,127.24,126.50,126.27,120.77,120.03,119.67,112.66,100.93,25.57;HR-ESI-MS:m/z＝278.12878[M+H]⁺,calcd for C₁₇H₁₆ON₃:278.12879.

EXAMPLE 33 preparation of Compound 33

(E) 3- (3- (1H-indol-6-yl) pyridin-4-yl) acrylic acid (132 mg,0.5 mmol), morpholine (52 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) dissolved in 10mL dichloromethane and stirred at room temperature for about 6H. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (50 mg, yield 30％).¹H NMR(400MHz,CDCl₃)δ8.69(s,1H),8.60(s,1H),8.54(s,1H),7.70(d,J＝8.0Hz,1H),7.64(d,J＝15.6Hz,1H),7.52(d,J＝5.2Hz,1H),7.33(s,1H),7.29-7.27(m,1H),7.05(d,J＝8.0Hz,1H),6.83(d,J＝15.6Hz,1H),6.59(s,1H),3.71-3.62(m,4H),3.64-3.59(m,2H),3.50-3.44(m,2H);¹³C NMR(125MHz,CDCl₃)δ164.75,151.19,147.62,141.21,139.18,138.24,135.82,129.72,127.76,125.55,122.41,121.75,120.94,120.79,112.47,102.50,66.74,66.66,46.26,42.39;HR-ESI-MS:m/z＝334.15472[M+H]⁺,calcd for C₂₀H₂₀O₂N₃:334.15500.

EXAMPLE 34 preparation of Compound 34

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (1228 mg,4.8 mmol), 1-methylindole-5-boronic acid (962mg,5.5mmol),Na₂CO₃(2035mg,19.2mmol),(PPh₃)₂PdCl₂(168mg,0.24mmol), was dissolved in a mixed solution of toluene, ethanol and water (volume ratio=1:1:0.2), and the mixed system was purged 5 times with nitrogen. Then the mixture is heated and refluxed for 12 hours in an oil bath under the protection of nitrogen. After the reaction, the solvent is distilled off, ethyl acetate is added, distilled water is used for washing, and the organic layer is combined and concentrated to obtain a solid crude product. The crude product was chromatographed on a silica gel column (petroleum ether: ethyl acetate=2:1 elution) to give a white solid (1000 mg, yield 71％).¹H NMR(400MHz,DMSO-d₆)δ8.64(s,1H),8.59(d,J＝5.2Hz,1H),7.86(d,J＝5.2Hz,1H),7.59(d,J＝8.4Hz,1H),7.55(d,J＝1.6Hz,1H),7.54(d,J＝16.0Hz,1H),7.44(d,J＝3.2,0.8Hz,1H),7.12(dd,J＝8.4,1.6Hz,1H),6.84(d,J＝16.0Hz,1H),6.51(dd,J＝3.2,0.8Hz,1H),4.14(q,J＝7.2Hz,2H),3.86(s,3H),1.20(t,J＝7.2Hz,3H);HR-ESI-MS:m/z＝307.14426[M+H]⁺,calcd for C₁₉H₁₉O₂N₂:307.14410.

EXAMPLE 35 preparation of Compound 35

(E) Ethyl-3- (3- (1-methyl-1H-indol-5-yl) pyridin-4-yl) acrylate (313 mg,2.0 mmol) is dissolved in 20ml absolute ethanol, 4ml of 1M aqueous NaOH solution is added dropwise and the mixture is stirred at room temperature for 6H. After the completion of the reaction, the solvent was distilled off, dissolved in distilled water, washed with ethyl acetate, and the aqueous layer was adjusted to pH 5 with 1M aqueous HCl to give a pale yellow solid (334 mg, yield) 60％).¹H NMR(400MHz,DMSO-d₆)δ8.61(s,1H),8.57(d,J＝5.2Hz,1H),7.81(d,J＝5.2Hz,1H),7.58(d,J＝8.4Hz,1H),7.53(d,J＝1.2Hz,1H),7.44(d,J＝16.0Hz,1H),7.43(d,J＝3.2Hz,1H),7.11(d,J＝8.4,Hz,1H),6.72(d,J＝16.0Hz,1H),6.51(dd,J＝3.2,1.2Hz,1H),3.86(s,3H);¹³C NMR(125MHz,DMSO-d₆)δ167.08,151.06,147.85,138.98,137.65,135.87,130.68,128.04,126.63,122.90,122.90,121.53,121.53,120.03,109.81,100.60,32.51;HR-ESI-MS:m/z＝279.11383[M+H]⁺,calcd for C₁₇H₁₅O₂N₂:279.11280.

EXAMPLE 36 preparation of Compound 36

(E) 3- (3- (1-methyl-1H-indol-5-yl) pyridin-4-yl) acrylic acid (139 mg,0.5 mmol), methylamine hydrochloride (40.5 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) in 10mL of dichloromethane and stirring at room temperature for about 6H. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (130 mg, yield 90％).¹H NMR(400MHz,DMSO-d₆)δ8.65(s,1H),8.62(d,J＝5.2Hz,1H),8.20(q,J＝4.4Hz,1H),7.70(d,J＝5.2Hz,1H),7.58(d,J＝8.4Hz,1H),7.54(d,J＝2.0Hz,1H),7.44(d,J＝3.2Hz,1H),7.35(d,J＝15.6Hz,1H),7.11(dd,J＝8.4,2.0Hz,1H),6.81(d,J＝15.6Hz,1H),6.51(dd,J＝2.8,0.8Hz,1H),3.86(s,3H),2.67(d,J＝4.4Hz,3H);¹³C NMR(125MHz,DMSO-d₆)δ164.44,149.85,146.66,141.01,137.93,135.92,134.40,130.71,128.01,126.98,126.32,122.86,121.54,120.12,109.84,100.63,32.51,25.58;HR-ESI-MS:m/z＝292.14410[M+H]⁺,calcd for C₁₈H₁₈ON₃:292.14444.

EXAMPLE 37 preparation of Compound 37

(E) 3- (3- (1-methyl-1H-indol-5-yl) pyridin-4-yl) acrylic acid (139 mg,0.5 mmol), morpholine (52 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) were dissolved in 10mL dichloromethane and the reaction stirred at room temperature for about 6H. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (110 mg, yield 63％).¹H NMR(400MHz,CDCl₃)δ8.70(s,1H),8.62(d,J＝4.8Hz,1H),7.60(d,J＝15.6Hz,1H),7.56-7.54(m,2H),7.40(d,J＝8.4Hz,1H),7.14-7.12(m,2H),6.85(d,J＝15.6Hz,1H),6.52(dd,J＝3.2,0.8Hz,1H),3.84(s,3H),3.69-3.64(m,4H),3.64-3.60(m,2H),3.52-3.48(m,2H);¹³C NMR(125MHz,CDCl₃)δ164.68,151.09,147.31,141.81,139.05,138.78,136.41,129.97,128.64,126.96,123.46,122.48,122.14,120.86,109.52,101.38,66.7,66.69,46.28,42.38,33.00;HR-ESI-MS:m/z＝348.16931[M+H]⁺,calcd for C₂₁H₂₂O₂N₃:348.17065.

EXAMPLE 38 preparation of Compound 38

(E) 3- (3- (quinolin-6-yl) pyridin-4-yl) acrylic acid (138 mg,0.5 mmol), methylamine hydrochloride (40.5 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) were dissolved in 10mL of dichloromethane and the reaction stirred at room temperature for about 6h. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (80 mg, yield 55％).¹H NMR(400MHz,DMSO-d₆)δ8.99(dd,J＝4.4,1.6Hz,1H),8.69(s,1H),8.68(d,J＝5.2Hz,1H),8.45(dd,J＝8.4,1.2Hz,1H),8.20(q,J＝4.8Hz,1H),8.14(d,J＝8.8Hz,1H),8.03(d,J＝1.6Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),7.69(d,J＝5.2Hz,1H),7.62(dd,J＝8.4,4.4Hz,1H),7.30(d,J＝16.0Hz,1H),6.81(d,J＝16.0Hz,1H),2.65(d,J＝4.8Hz,3H);¹³C NMR(125MHz,DMSO-d₆)δ164.34,151.17,150.80,149.10,146.93,139.96,136.22,135.43,134.28,133.83,131.10,129.01,128.92,127.64,127.31,122.04,119.86,25.57;HR-ESI-MS:m/z＝290.12762[M+H]⁺,calcd for C₁₈H₁₆ON₃:290.12879.

EXAMPLE 39 preparation of Compound 39

(E) 3- (3- (quinolin-6-yl) pyridin-4-yl) acrylic acid (138 mg,0.5 mmol), morpholine (52 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) was dissolved in 10mL of dichloromethane and the reaction stirred at room temperature for about 6h. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (80 mg, yield 46％).¹H NMR(400MHz,DMSO-d₆)δ8.99(dd,J＝4.4,2.0Hz,1H),8.70(s,1H),8.69(d,J＝5.2Hz,1H),8.44(dd,J＝8.8,2.0Hz,1H),8.15(d,J＝8.4Hz,1H),8.03(s,1H),8.03(d,J＝8.4Hz,1H),7.75(dd,J＝8.8,2.0Hz,1H),7.63(dd,J＝8.4,4.4Hz,1H),7.50(d,J＝15.6Hz,1H),7.39(d,J＝15.6Hz,1H),3.72-3.69(m,2H),3.61-3.55(m,4H),3.51-3.47(m,2H);¹³C NMR(125MHz,DMSO-d₆)δ163.48,151.19,150.77,148.89,146.90,139.87,136.69,136.20,135.52,134.31,131.14,129.00,128.96,127.61,123.21,122.06,120.35,66.21,65.92,45.47,41.96;HR-ESI-MS:m/z＝346.15396[M+H]⁺,calcd for C₂₁H₂₀O₂N₃:346.15500.

EXAMPLE 40 preparation of Compound 40

(E) 3- (3- (quinolin-7-yl) pyridin-4-yl) acrylic acid (138 mg,0.5 mmol), methylamine hydrochloride (40.5 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) were dissolved in 10mL of dichloromethane and the reaction stirred at room temperature for about 6h. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (90 mg, yield 62％).¹H NMR(400MHz,DMSO-d₆)δ8.99(dd,J＝4.0,1.6Hz,1H),8.71(s,1H),8.69(d,J＝5.2Hz,1H),8.50-8.46(m,1H),8.21(q,J＝5.2Hz,1H),8.14(d,J＝8.4Hz,1H),8.01-7.99(m,1H),7.70(d,J＝5.2Hz,1H),7.64-7.61(m,2H),7.32(d,J＝15.6Hz,1H),6.81(d,J＝15.6Hz,1H),2.66(d,J＝4.4Hz,3H);¹³C NMR(125MHz,DMSO-d₆)δ164.35,151.27,150.73,149.15,147.24,139.98,137.27,135.86,135.35,133.95,129.57,128.39,128.12,127.33,127.20,122.00,119.98,25.57;HR-ESI-MS:m/z＝290.12878[M+H]⁺,calcd for C₁₈H₁₆ON₃:290.12879.

EXAMPLE 41 preparation of Compound 41

(E) 3- (3- (quinolin-7-yl) pyridin-4-yl) acrylic acid (138 mg,0.5 mmol), morpholine (52 mg,0.6 mmol), HATU (228 mg,0.6 mmol), DIEA (129 mg,1.0 mmol) was dissolved in 10mL of dichloromethane and the reaction stirred at room temperature for about 6h. After the reaction, the reaction solution was washed with saturated sodium carbonate solution, 0.5N hydrochloric acid solution, and saturated brine, and the organic layer was concentrated to give a crude product, which was subjected to silica gel column chromatography to give a pale yellow solid (112 mg, yield 65％).¹H NMR(400MHz,DMSO-d₆)δ8.99(dd,J＝4.4,1.6Hz,1H),8.71(d,J＝0.8Hz,1H),8.69(dd,J＝5.2,0.8Hz,1H),8.50-8.46(m,1H),8.14(d,J＝8.4Hz,1H),8.02(d,J＝5.2Hz,1H),8.00-7.99(m,1H),7.63(dd,J＝8.4,3.2Hz,1H),7.62(d,J＝8.4Hz,1H),7.50(d,J＝15.6Hz,1H),7.42(d,J＝15.6Hz,1H),3.75-3.67(m,2H),3.63-3.53(m,4H),3.52-3.45(m,2H);¹³C NMR(125MHz,DMSO-d₆)δ163.47,151.30,150.70,148.94,147.20,139.87,137.31,136.76,135.87,135.44,129.62,128.40,128.17,127.17,123.24,122.01,120.45,66.21,65.91,45.47,41.96;HR-ESI-MS:m/z＝346.15503[M+H]⁺,calcd for C₂₁H₂₀O₂N₃:346.15500.

EXAMPLE 42 preparation of Compound 42

(1) (E) -3- (3- (1H-indazol-5-yl) pyridin-4-yl) acrylic acid ethyl ester

(E) Ethyl 3- (3-bromopyridin-4-yl) acrylate (1228 mg,4.8 mmol), 1H-indole-5-boronic acid pinacol ester (1342mg,5.5mmol),Na₂CO₃(2035mg,19.2mmol),(PPh₃)₂PdCl₂(168mg,0.24mmol), was dissolved in a mixed solution of toluene, ethanol and water (volume ratio=1:1:0.2), and the mixed system was purged 5 times with nitrogen. Then the mixture is heated and refluxed for 12 hours in an oil bath under the protection of nitrogen. After the reaction, the solvent is distilled off, ethyl acetate is added, distilled water is used for washing, and the organic layer is combined and concentrated to obtain a solid crude product. The crude product was chromatographed on a column of silica gel (petroleum ether: ethyl acetate=2:1 to give a white solid (1014 mg, 72% yield).

(2) (E) -3- (3- (1H-indazol-5-yl) pyridin-4-yl) acrylic acid

(E) Ethyl 3- (3- (1H-indazol-5-yl) pyridin-4-yl) acrylate (586 mg,2.0 mmol) was dissolved in 20ml absolute ethanol, 4ml of 1M aqueous NaOH solution was added dropwise thereto, and the mixture was stirred at room temperature for 6 hours. After the completion of the reaction, the solvent was distilled off, dissolved in distilled water, washed with ethyl acetate, and the aqueous layer was adjusted to pH 5 with 1M aqueous HCl to precipitate a pale yellow solid (329 mg, yield 62%).

(3) (E) -3- (3- (1H-indazol-5-yl) pyridin-4-yl) acryloyl-3-methoxycyclobutylamine

(E) Reaction of 3- (3- (1H-indazol-5-yl) pyridin-4-yl) acrylic acid with 3-methoxycyclobutylamine hydrochloride, and the method and operation are the same as those of Compound 4 .¹H NMR(400MHz,CDCl₃)δ10.92(s,1H),8.66(s,1H),8.62(d,J＝5.2Hz,1H),8.07(d,J＝1.2Hz,1H),7.68(d,J＝16.0Hz,1H),7.67-7.68(m,1H),7.51(d,J＝5.6Hz,1H),7.46-7.49(m,1H),7.28(d,J＝1.6Hz,1H),6.57(d,J＝15.6Hz,1H),4.36-4.42(m,1H),4.20-4.31(m,2H),4.06-4.16(m,1H),3.95-4.00(m,1H),3.32(s,3H);¹³C NMR(100MHz,DMSO-d₆)δ164.25,151.05,148.29,139.71,139.37,136.81,135.75,133.80,128.21,127.95,122.98,122.10,121.71,120.37,110.37,68.40,56.81,55.38,54.65;HR-ESI-MS:m/z＝335.1503[M+H]⁺,calcd for C₁₉H₁₉O₂N₄:335.1503.

EXAMPLE 43 preparation of Compound 43

(1) Synthesis of ethyl 3- (3- (naphthalen-2-yl) pyridin-4-yl) propionate

(E) Ethyl 3- (3- (naphthalen-2-yl) pyridin-4-yl) acrylate (460 mg,1.53 mmol) was dissolved in 5mL of ethanol, pd/C (44 mg, 10%) was added, the atmosphere was replaced three times with an atmospheric hydrogen balloon, and the reaction was then stirred at room temperature for about 3 hours. After the reaction was completed, palladium on carbon was filtered off with celite, the filter cake was washed with methanol several times, and the filtrate was concentrated under reduced pressure to give a white solid (443 mg, yield 95%).

(2) Synthesis of 3- (3- (naphthalen-2-yl) pyridin-4-yl) propionic acid

Ethyl 3- (3- (naphthalen-2-yl) pyridin-4-yl) propionate (610 mg,2.0 mmol) was dissolved in 20ml absolute ethanol, 4ml of 1M aqueous NaOH was added dropwise, and the mixture was stirred at room temperature for 6h. After the completion of the reaction, the solvent was distilled off, dissolved in distilled water, washed with ethyl acetate, and the aqueous layer was adjusted to pH 5 with 1M aqueous HCl to give a pale yellow solid (338 mg, yield) 61％).¹H NMR(400MHz,DMSO-d₆)δ12.21(br s,1H),8.52(d,J＝5.2Hz,1H),8.46(s,1H),8.03(d,J＝8.4Hz,1H),8.01-7.98(m,2H),7.94(s,1H),7.58(q,J＝3.2Hz,2H),7.55(d,J＝8.4Hz,1H),7.43(d,J＝4.8Hz,1H),2.87(t,J＝8.0Hz,2H),2.47(t,J＝8.0Hz,2H);¹³C NMR(125MHz,DMSO-d₆)δ173.37,149.82,148.61,147.11,137.04,134.91,132.86,132.16,128.05,128.05,127.94,127.63,127.33,126.55,126.46,123.54,33.27,27.09;HR-ESI-MS:m/z＝278.11853[M+H]⁺,calcd for C₁₈H₁₆O₂N:278.11756.

EXAMPLE 44 preparation of Compound 44

(E) The synthetic method and the operation are the same as those of the compound 43, namely, the 3- (3- (naphthalene-2-yl) pyridine-4-yl) methyl acrylate is obtained through reduction reaction .¹H NMR(400MHz,DMSO-d₆)δ8.53(d,J＝5.2Hz,1H),8.46(s,1H),8.03(d,J＝8.4Hz,1H),8.01-7.98(m,2H),7.93(s,1H),7.58(d,J＝8.8Hz,1H),7.58(d,J＝2.8Hz,1H),7.55(dd,J＝8.4,1.6Hz,1H),7.43(d,J＝5.2Hz,1H),3.49(s,3H),2.90(t,J＝8.0Hz,2H),2.58(t,J＝8.0Hz,2H);¹³C NMR(125MHz,DMSO-d₆)δ172.14,149.74,148.52,146.60,136.91,134.70,132.73,132.03,127.95,127.92,127.83,127.50,127.20,126.44,126.36,123.39,51.27,32.76,26.75;HR-ESI-MS:m/z＝292.13425[M+H]⁺,calcd for C₁₉H₁₈O₂N:292.13321.

EXAMPLE 45 preparation of Compound 45

(E) The synthetic method and the operation are the same as those of the compound 43, namely, the compound is obtained by the reduction reaction of the-3- (3- (naphthalene-2-yl) pyridine-4-yl) acrylamide methylamine .¹H NMR(500MHz,DMSO-d₆)δ8.52(d,J＝4.0Hz,1H),8.45(s,1H),8.02(d,J＝6.8Hz,1H),8.01-7.97(m,2H),7.93(d,J＝0.8Hz,1H),7.74(d,J＝4.0Hz,1H),7.58(d,J＝6.8Hz,1H),7.58(dd,J＝2.4,0.8Hz,1H),7.54(dd,J＝6.8,1.6Hz,1H),7.38(d,J＝4.0Hz,1H),2.85(t,J＝6.4Hz,2H),2.49(d,J＝4.0Hz,3H),2.31(t,J＝6.4Hz,2H);HR-ESI-MS:m/z＝291.15018[M+H]⁺,calcd for C₁₉H₁₉ON₂:291.14919.

EXAMPLE 46 preparation of Compound 46

(E) The method and operation of the reduction reaction of the-3- (3- (naphthalene-2-yl) pyridine-4-yl) acryloylmorpholine are the same as that of the compound 43 .¹H NMR(400MHz,DMSO-d₆)δ8.52(d,J＝5.2Hz,1H),8.45(s,1H),8.03(d,J＝8.4Hz,1H),8.00(d,J＝10.8Hz,1H),8.00(d,J＝3.2Hz,1H),7.94(d,J＝0.8Hz,1H),7.54(d,J＝10.8Hz,1H),7.54(d,J＝3.2,0.8Hz,1H),7.55(dd,J＝8.4,1.6Hz,1H),7.46(d,J＝5.2Hz,1H),3.43(t,J＝4.8Hz,2H),3.34-3.30(m,4H),3.19(t,J＝4.8Hz,2H),2.88-2.81(m,2H),2.57-2.51(m,2H);¹³C NMR(125MHz,DMSO-d₆)δ169.54,149.74,148.58,147.68,137.07,134.98,132.85,132.14,128.10,128.03,127.95,127.63,127.46,126.59,126.49,123.86,65.99,65.90,45.15,41.39,32.21,27.39;HR-ESI-MS:m/z＝347.17636[M+H]⁺,calcd for C₂₂H₂₃O₂N₂:347.17540.

EXAMPLE 47 preparation of Compound 47

(E) Ethyl 3- (3- (naphthalen-2-yl) pyridin-4-yl) acrylate (30 mg,0.1 mmol), hydrazine hydrate (25 mg,5 mmol) was dissolved in 5mL ethanol and the reaction was heated to reflux and reacted for about 4h. After completion of the reaction, the solvent was distilled off, and washed with a small amount of ethyl acetate to give a white flocculent solid (15 mg, yield) 51％).¹H NMR(400MHz,DMSO-d₆)δ8.93(s,1H),8.50(d,J＝5.2Hz,1H),8.44(s,1H),8.01(d,J＝8.8Hz,1H),7.96-7.99(m,2H),7.91(d,J＝1.2Hz,1H),7.55-7.59(m,2H),7.53(dd,J＝8.4,1.6Hz,1H),7.37(d,J＝4.8Hz,1H),4.09(s,2H),2.82-2.86(m,2H),2.25-2.29(m,2H);¹³C NMR(100MHz,DMSO-d₆)δ170.20,149.80,148.52,147.38,136.96,134.84,132.82,132.13,128.05,128.02,127.87,127.57,127.36,126.49,126.40,123.46,33.10,27.57;HR-ESI-MS:m/z＝292.1444[M+H]⁺,calcd for C₁₈H₁₈ON₃:292.1444.

Pharmacological experiments:

Example 1 in vitro kinase inhibitory Activity assay

The compound was diluted 100-fold in DMSO solution, mixed with buffer (50mM HEPES pH 7.5,0.01%BRIJ-35,10mM MgCl ₂, 1mM EGTA), a mixture of kinase and antibody, and Tracer, shaken for 30s, and then incubated at room temperature for 60min. The Emission Ratio (ER, AF647 Emission (665 nm) to Europium Emission (615 nm) Ratio) was then read on a microplate reader and analyzed. In the IC ₅₀ test, compounds were diluted in 3-fold gradients for a total of 10 concentrations. A blank control was set up in the test, the positive control being referenced to sorafenib. The results are given as percentages of "difference between ER of the blank and ER of the test sample" and "difference between ER of the blank and ER of the positive control", with greater percentages indicating better binding of the compound to the kinase. Table 1 shows the results of the in vitro inhibitory activity of CDK8 kinases of the compounds of the invention.

TABLE 1

ND Not Determined, untested.

Example 2 in vitro kinase Selective Activity assay

The invention selects 13 representative effective inhibitors to selectively test 12 kinases. Of which 4 are CDK subtypes including CDK2, CDK6, CDK7 and CDK9, and the other 8 are Aurora A, BRAF, EGFR, FGFR1, FLT3, JAK1, PDGFRA and GSK3A. The kinase selectivity of the 13 compounds selected according to the invention was determined at a concentration of 10. Mu.M. As shown in table 2 and fig. 1, each of the 13 compounds of the present invention showed selective inhibition of CDK 8.

TABLE 2 inhibition of 13 kinases by 13 Compounds of the invention

CDKs: cyclin dependent kinase (CYCLIN DEPENDENT KINASE), BRAF: sarcoma filtered viral oncogene homolog B1 (v-raf murine sarcomaviral oncogene homolog B1), aurora A: aurora kinase A (Aurora kinase A), EGFR: epidermal growth factor receptor (EPITHELIAL GROWTH FACTOR RECEPTOR), PDGFRα: platelet derived growth factor receptor α (Platelet-derived growth factor receptor α), JAK1: janus kinase 1 (Janus kinase 1), FGFR1: fibroblast growth factor receptor 1 (Fibroblast Growth Factor Receptor-1), GSK3α: glycogen synthase kinase 3α (glycogen SYNTHASE KINASE 3α), FLT3: FMS-like tyrosine kinase 3 (FMS-like tyrosine kinase).

Example 3 in vitro anti-tumor cell Activity assay

The invention adopts SRB method to test the cytotoxic activity of 31 compounds with better molecular level activity, and selects cytoma strains of A549, MDA-MB-231, KB-VIN and MCF-7 respectively, and the test results are shown in Table 2. First, the trypsin-treated cell suspension is added to a 96-well plate with a cell density of 4000-11000 cells/well. Then, diluted compound solution was added, mixed with the cells, and after three days of incubation, the adhered cells were fixed with 10% ice-bath trichloroacetic acid and stained with 0.04% sulfonylrhodamine B (sulforhodamine B). Absorbance at 515nm was measured using a microplate reader (ELx 800,800, bioTek). IC ₅₀ values were obtained from the results of three independent experiments by de-averaging.

TABLE 2

Claims

1. A pyridine compound represented by the following general formula (IA) and a pharmaceutically acceptable salt thereof,

in,

_R1 is selected from halogen, substituted or unsubstituted phenyl, naphthyl, substituted or unsubstituted indolyl, quinolyl, substituted or unsubstituted indazolyl; the substituents are mono- or poly-substituted groups, each of which is independently selected from halogen, C1 _- _C3 alkyl, C1 _- _C3 alkoxy;

R ₂ is selected from C ₁ -C ₃ alkylamino, morpholinyl, and N-methylpiperazinyl.

2. A compound having the following structure and a pharmaceutically acceptable salt thereof, characterized in that the compound is selected from:

3. The method for preparing the compound according to claim 1, characterized in that it comprises the following steps:

The compound of general formula (IA) is obtained from 3-bromoisonicotinaldehyde as the starting material:

_R1 and _R2 are as defined in claim 1.

4. A pharmaceutical composition comprising an effective dose of any one of the compounds according to any one of claims 1 to 2 and a pharmaceutically acceptable carrier thereof.

5. The pharmaceutical composition according to claim 4, characterized in that the pharmaceutical composition is selected from tablets, capsules, pills, injections, sustained-release preparations, controlled-release preparations or various microparticle delivery systems.

6. Use of the compound according to any one of claims 1 to 2 and a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and/or treating CDK8-related diseases.

7. The use according to claim 6, characterized in that the CDK8-related disease is selected from tumors and immune-related diseases.

8. The use according to claim 7, characterized in that the tumor includes colorectal cancer, breast cancer, gastric cancer, ovarian cancer, and melanoma.