CN114075189A

CN114075189A - Five-membered heterocyclic acene ring compound and preparation method and medical application thereof

Info

Publication number: CN114075189A
Application number: CN202010803248.7A
Authority: CN
Inventors: 秦引林; 苏梅; 王德忠
Original assignee: Jiangsu Carephar Pharmaceutical Co ltd; Nanjing Carephar Shenghui Pharmaceutical Co ltd
Current assignee: Jiangsu Carephar Pharmaceutical Co ltd
Priority date: 2020-08-11
Filing date: 2020-08-11
Publication date: 2022-02-22

Abstract

本发明公开一种五元杂环并苯环类化合物及其制备方法和医药用途，涉及结构如式(I)所示的化合物或其立体异构体、互变异构体或其药学上可接受的盐或其溶剂合物或者前药，本发明还提供本发明式I的化合物在用于制备预防或治疗JAK激酶相关疾病药物中的用途。

The invention discloses a five-membered heterocyclic acene ring compound, a preparation method and medical use thereof, and relates to a compound whose structure is shown in formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable compound thereof. The accepted salt or solvate or prodrug thereof, the present invention also provides the use of the compound of formula I of the present invention in the preparation of a medicament for preventing or treating JAK kinase-related diseases.

Description

Five-membered heterocyclic acene ring compound and preparation method and medical application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a novel five-membered heterocyclic acene ring compound as well as a preparation method and application thereof.

Background

Chondrocytes are the major cellular component of avascular tissue, with chondrocytes in normal articular cartilage accounting for about 5% of the tissue volume, while extracellular matrix accounts for the remaining 95% of the tissue. Chondrocytes secrete matrix components, mainly proteoglycans and collagen, which in turn provide the chondrocytes with an environment suitable for their survival under mechanical stress. In cartilage, type II collagen forms a fiber-like structure with type IX collagen, providing cartilage with great mechanical strength. Proteoglycans can absorb water and are responsible for the elastic and shock absorbing properties of cartilage. One of the functional roles of cartilage is to provide smooth articulation of bone portions to one another. Thus, the loss of articular cartilage causes bones to rub against each other, resulting in pain and loss of motion. In inflammatory arthritis, such as rheumatoid arthritis, cartilage degradation is caused by proteases (collagenases) secreted by inflamed tissues, such as inflamed synovium. Chondrocytes in damaged cartilage often show a decrease in cartilage synthesis activity and/or an increase in cartilage degeneration activity, which is a major disease marker for the development of rheumatoid arthritis and osteoarthritis. Rheumatoid Arthritis (RA) is a chronic degenerative disease of the joints characterized by inflammation and destruction of joint structures, possibly leading to malfunction of joint function causing substantial disability and pain, and even premature death. Therefore, the purpose of RA treatment is not only to delay disease, relieve pain, and protect against joint destruction.

JAK kinases belong to the Janus kinase (JAK) family of cytoplasmic tyrosine kinases, which are involved in cytokine receptor-mediated intracellular signal transduction. The JAK kinase family includes 4 members: JAK1, JAK2, JAK3 and TYK 2. JAKs recruit cytokine receptors, bind cytokines, and subsequently dimerize the cytokine receptors and shared receptor subunits. JAKs are then activated by autophosphorylation and/or transphosphorylation of another JAK, resulting in receptor phosphorylation and recruitment and phosphorylation of members of Signal Transducers and Activators of Transcription (STATs). Phosphorylated SATA dimerizes and translocates into the nucleus where they bind to enhancer regions of cytokine response genes. JAKs play an important role in the regulation of biological response functions of multiple cytokine receptor families. JAK1 knockout mice have early postnatal lethal factor phenotypes and the nervous system is also compromised, leading to congenital defects in young mice. The research shows that the JAK1 gene knockout mouse has secretion defects of thymocytes and B cells, and the JAK1 gene knockout tissue has obviously weakened response to IL-6 and IL-10.

The association of JAKs with autoimmune diseases has been established clinically, with mutations in JAKs and the upstream signaling component r-c receptor chain and IL7 receptor detected in about 70% of severe combined immunodeficiency cases.

Conditions to which the JAK family member JAK2 is clinically implicated include myeloproliferative disorders, cancer, particularly leukemias such as acute myelogenous leukemia, acute lymphocytic leukemia, or related diseases with solid tumors such as uterine leiomyosarcoma, prostate cancer, and the like.

Disclosure of Invention

The invention provides a preparation method and medical application of a novel five-membered heterocyclic acene ring compound.

More specifically, the invention relates to a compound with a structure shown as a formula (I) or a stereoisomer, a tautomer or a pharmaceutically acceptable salt or a solvate or a prodrug thereof:

wherein:

a is independently selected from S, N, N (Me);

b is independently selected from substituted or unsubstituted C₃-C₆Cycloalkyl, a 5-to 6-membered heteroaryl ring having 1 to 2 ring heteroatoms independently selected from N, O and S, wherein said 5-to 6-membered heteroaryl ring is optionally substituted with C independently selected from halogen, fluoro or non-fluoro₁-C₆Alkyl, fluoro or non-fluoro C₁-C₆Alkoxy, fluoro or non-fluoro C₁-C₆One or more substituents of the alkylamino group;

d is independently selected from C, N;

L₁is absent or independently selected from the group consisting of a single bond, -CH₂-，-(CH₂)_xO(CH₂)_y-,-(CH₂)_xNH(CH₂)_y-,-CH₂O-,-C(O)-,-CON(R₄)-,-CH₂N(R₄)-,-CONH(CH₂)_y-,-N(R₄)-,-SO₂N(R₄)-,-S(O)₂-,-N(Me)-；

R₄Independently selected from H, C₁-C₆Alkyl, substituted C₁-C₆Alkyl radical, C₁-C₃Ether C₁-C₃Alkyl, methylsulfonyl;

R₁independently selected from H, -NH₂-COOH, substituted or unsubstituted C₁-C₆Alkyl, acyl, substituted or unsubstituted acylamino, substituted or unsubstituted C₁-C₆Alkoxy, halogen, hydroxy, substituted or unsubstituted C₁-C₃An ester group, a substituted or unsubstituted heteroaryl group;

R₃independently selected from H, substituted or unsubstituted: sulfonyl, sulfonamide, 5-to 6-membered heterocycle having at least one heteroatom and optionally a second ring heteroatom independently selected from N and S, substituted or unsubstituted C₁-C₆Alkane, substituents independently selected from 5 to6 membered heteroaryl and methoxy substituted, substituted or unsubstituted C₃-C₇Cycloalkyl, substituted or unsubstituted 4-7 membered heterocycloalkyl, substituted or unsubstituted 5-7 membered heteroaryl.

m is 0, 1,2, 3; n is 0, 1,2, 3; t is 0, 1,2, 3.

In one embodiment, characterized in that B is selected from the following substituted or unsubstituted groups: cyclopropane, cyclobutane, cyclopentane, cyclohexane, pyrazolyl, pyridyl, thiophene, thiazole, 1,3, 4-thiadiazole, oxazole, 1,3, 4-oxadiazole, furan, pyrrole, 3-pyrroline, 2-pyrazoline, imidazole, 1,2, 3-triazole, 1,2, 4-triazole, pyran, pyridazine, pyrimidine, pyrazine; the substituent is selected from halogen and C₁-C₃Alkyl radical, C₁-C₃A haloalkyl group; preferably, B is selected from the following substituted or unsubstituted groups: cyclopropane, cyclobutane, pyrazolyl, pyridyl, imidazolyl; the substituent is selected from F, Cl, Br, methyl, ethyl, propyl, isopropyl and trifluoromethyl.

In one embodiment, D is C or N.

In one embodiment, n is 1.

In one embodiment, R₁Independently selected from H, -NH₂-COOH, hydroxy, halogen, substituted or unsubstituted C₁-C₃Alkyl, substituted or unsubstituted C₁-C₃Acyl, substituted or unsubstituted C₁-C₃Acylamino, substituted or unsubstituted C₁-C₃Alkoxy, substituted or unsubstituted C₁-C₃An ester group; in a preferred embodiment, R₁Independently selected from H, -NH₂-COOH, hydroxy, halogen, methyl, ethyl, propyl, isopropyl, formylamino, carbomethoxy.

In one embodiment, m is 0 or 1, more preferably, m is 0.

In one embodiment, L₁Is absent or independently selected from the group consisting of a single bond, -CH₂-，-(CH₂)_xO(CH₂)_y-,-(CH₂)_xNH(CH₂)_y-,-CONH(CH₂)_y-,-N(R₄)-；R₄Independently selected from H, methyl, ethyl, propyl, ethyl methyl ether, methyl ether, ethyl ether; x or y is independently selected from 0, 1 or 2. In a more preferred embodiment, L₁Is absent or independently selected from the group consisting of a single bond, -CH₂-，-(CH₂)_xO(CH₂)_y-,-(CH₂)_xNH(CH₂)_y-,-CONH(CH₂)_y-,-N(R₄)-；R₄Independently selected from H, methyl, ethyl methyl ether; x or y is independently selected from 0, 1 or 2.

In one embodiment, t is 1.

In one embodiment, R₃Independently selected from H, substituted or unsubstituted: sulfonyl, sulfonamide, thiomorpholine 1, 1-dioxide, piperidine, pyrazole, thiazole, imidazole, 1,3, 4-thiadiazole, piperazine, morpholine, pyrimidine, pyrazine, thiophene, oxazole, 1,3, 4-oxadiazole, furan, pyrrole, 3-pyrroline, 2-pyrazoline, 1,2, 3-oxazole, 1,2, 3-triazole, 1,2, 4-triazole, pyran; the substituent is selected from H, halogen and C₁-C₃Alkyl radical, C₁-C₃Haloalkyl, hydroxy C₁-C₃Alkyl radical, C₃-C₆Cycloalkyl, ethyl methyl ether, methyl ether, ethyl ether; preferably, R₃Independently selected from H, substituted or unsubstituted: sulfonyl, sulfonamide, thiomorpholine 1, 1-dioxide, piperidine, pyrazole, thiazole, imidazole, 1,3, 4-thiadiazole, piperazine, morpholine, pyrimidine, thiophene, oxazole, 1,3, 4-oxadiazole, furan, pyrrole, 3-pyrroline, 2-pyrazoline, 1,2, 3-oxazole, 1,2, 3-triazole, 1,2, 4-triazole; the substituent is selected from H, halogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, hydroxyethyl, hydroxymethyl, cyclopropane, cyclobutane, ethyl methyl ether, methyl ether, ethyl ether.

The present invention also provides compounds represented by the following structures or stereoisomers, tautomers, or pharmaceutically acceptable salts, or solvates, or prodrugs thereof:

the invention also provides methods for synthesizing the compounds of the invention and disclosed representative synthetic schemes and routes.

The present invention also provides a pharmaceutical composition comprising a compound of formula I as described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent or carrier.

The invention also provides the use of a compound of formula I according to the invention for the preparation of a medicament for the prevention or treatment of a JAK kinase-associated disease, in particular for the preparation of a medicament for the prevention and/or treatment of diseases involving cartilage degradation, bone and/or joint degradation, conditions involving inflammation or immune response, endotoxin-driven disease states, cancer and organ transplant rejection.

The invention also provides a method for the prevention and/or treatment of diseases involving cartilage degradation, bone and/or joint degradation, conditions involving inflammation or immune response, endotoxin-driven disease states, cancer and organ transplant rejection by administering a compound of the invention.

In one embodiment, examples of diseases related to cartilage degradation, bone and/or joint degradation, conditions related to inflammation or immune response, endotoxin-driven disease states, cancer, and organ transplant rejection described herein include, but are not limited to: osteoarthritis, crohn's disease, rheumatoid arthritis, psoriasis, allergic airway diseases (e.g., asthma, rhinitis), juvenile idiopathic arthritis, colitis, inflammatory bowel disease, endomycin driven disease states, diseases with impairment of cartilage turnover (e.g., diseases with anabolic stimulation of chondrocytes), congenital cartilage malformations, organ transplant rejection, diseases involving cartilage degeneration, joint degeneration, myeloproliferative disorders, leukemia (acute myeloid leukemia, acute lymphocytic leukemia), solid tumors (uterine leiomyosarcoma, prostate cancer), and the like.

Detailed Description

The present invention is further described with reference to the following examples, which are not intended to limit the scope of the present invention, and all simple modifications of the preparation method of the present invention based on the concept of the present invention are within the scope of the present invention. The following examples are experimental methods without specifying specific conditions, and generally follow the methods known in the art. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.

Example 1: preparation of Compound EXP-1

The synthetic route is as follows:

step 1.1 preparation of Compound 1

5-Nitrobenzothiophene (SM1) (100mg,0.558mmol) was dissolved in ethanol (3mL) and palladium on carbon (5mg, 10% purity) was added thereto under an argon atmosphere. The reaction solution was replaced several times with hydrogen and heated to 40 ℃ with continuous stirring under 30psi pressure for 16 hours. The reaction was filtered, and the filtrate was concentrated in vacuo to give 5-aminobenzothiophene (1) (100mg, yield 100%) as a yellow solid.

1H NMR(400MHz,CDCl₃):δ＝7.65(d,J＝8.4Hz,1H),7.39(d,J＝5.6Hz,1H),7.16(d,J＝5.2Hz,1H),7.12(d,J＝2.4Hz,1H),6.80(dd,J＝8.4and2.4 Hz,1H),3.62(brs,2H).

Step 1.2 preparation of Compound 2

5-aminobenzothiophene (1) (100mg,0.67mmol) and triethylamine (136mg,1.34mmol) were dissolved in dichloromethane (2mL) and cooled to 0 ℃. Methanesulfonyl chloride (92mg,0.804mmol) was added slowly and stirred at 0-20 ℃ for 2 hours. The reaction was diluted with ethyl acetate (10mL) and water (10mL) and the aqueous phase was extracted with ethyl acetate (10mL x 3). The organic phase is dried and filtered by anhydrous sodium sulfate, and the filtrate is dried by spinning to obtain a crude product. The crude product was chromatographed on silica gel column (eluent: 0% to 30% ethyl acetate/petroleum ether) to give N- [ benzothien-5-yl ] -N-methylsulfonylmethanesulfonamide (2) (100mg, yield 48.86%) as a white solid.

1H NMR(400MHz,CDCl₃):δ＝7.97(d,J＝8.4Hz,1H),7.84(d,J＝2.0Hz,1H),7.58(d,J＝5.2Hz,1H),7.40(d,J＝5.2Hz,1H),7.31(dd,J＝8.4and2.0 Hz,1H),3.46(s,6H).

Step 1.3 preparation of Compound 3

N- [ benzothien-5-yl ] -N-methylsulfonylmethanesulfonamide (2) (50mg,0.164mmol) was dissolved in tetrahydrofuran (0.5mL) and cooled to-75 ℃. N-butyllithium (2.5M n-hexane solution, 0.16mL) was added slowly at-75 ℃ under a nitrogen atmosphere. After stirring for 60 minutes, triisopropoxyl borate (46mg,245.59umol) was added to the reaction solution and stirred at-75 ℃ for 15 minutes. The temperature was slowly raised to 15-20 ℃ and stirring was continued for 16 hours. The reaction was quenched with 10mL of 10% hydrochloric acid solution and extracted with ethyl acetate (20 mL. times.3), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was spun dry to give [5- (methanesulfonamide) benzothien-2-yl ] boronic acid (3) (30mg, crude) as a brown oily solid which was used in the next step without purification.

Step 1.4 preparation of Compound EXP-1

[5- (Methanesulfonamide) benzothien-2-yl group]Boronic acid (3) (20mg,0.074mmol) and (5-bromo- [1,2, 4)]Triazole [1,5-a ]]Pyridin-2-amine) Cyclopropanecarboxamide (Int-1) (21mg,0.074mmol) was dissolved in dioxane (0.5mL) and water (0.1mL), and potassium carbonate (31mg,0.221mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (10.8mg,0.0148mmol) were added rapidly under nitrogen. The mixture was replaced with nitrogen three times, and then stirred at 90 ℃ for 2 hours. The reaction mixture was filtered, the filtrate was diluted with ethyl acetate (10mL) and water (10mL), the aqueous phase was extracted with ethyl acetate (10 mL. times.3), the organic phase was dried over anhydrous sodium sulfate and the crude product concentrated by filtration was purified by preparative HPLC (separation column: BostonPrime C18150: 30 mm. times.5 μm; mobile column: Bio-Rad;)Phase A Water (0.04% NH)₃H₂O+10mM NH₄HCO₃) And B is acetonitrile; gradient: b25-55%) and obtaining the white target product N- [5- [5- (methanesulfonamide) benzothiophen-2-yl][1,2,4]Triazole [1,5-a ]]Pyridin-2-yl]Cyclopropanecarboxamide (EXP-1) (3.7mg, yield 11% yield).

LCMS:t_R＝1.860min in 10-80CD_7min_220&254_Agilent.met,chromatography(Xbrige C18 30*2.1mm*3.5μm),MS(ESI)m/z＝428.1[M+H]⁺.

HPLC:t_R＝2.06min in 10-80CD_8min.met,chromatography(XBridge C18 2.1*50mm*5μm).

1H NMR(400MHz,DMSO-d₆):δ＝11.25(s,1H),9.87(s,1H),8.81(s,1H),8.05(d,J＝8.8Hz,1H),7.86-7.65(m,4H),7.34(d,J＝8.8Hz,1H),3.02(s,3H),2.03-1.94(m,1H),0.91-0.84(m,4H).

The example compounds of the following table 1 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 1]

Example 2: preparation of Compound EXP-4

The synthetic route is as follows:

step 2.1 preparation of Compound 4

4-Methanesulfonylaniline (SM2)0(1g,5.84mmol) was dissolved in ethanol (20mL) and iodine (1.48g,5.84mmol) and silver sulfate (2.00g,6.42mmol) were added. The reaction solution was stirred at 50 ℃ for 2 hours. The reaction was filtered and the filtrate was spin dried to give 2-iodo-4-methanesulfonyl-aniline (740mg, 43% yield) as a brown solid.

LCMS:t_R＝0.714min in 5-95AB_1.5min_220&254_Shimadzu.lcm chromatography(Merck RP18e 25-3mm),MS(ESI)m/z＝297.8[M+H]⁺.

Step 2.2 preparation of Compound 6

2-iodo-4-methanesulfonyl-aniline (4) (250mg,1.11mmol), N- [ 5-ethynyl- [1,2,4] triazolo [1,5-a ] pyrid-2-yl ] cyclopropanecarboxamide (5) (328mg,1.11mmol), cuprous iodide (42mg,0.221mmol), triethylamine (559mg,5.53mmol) and triphenylphosphine palladium dichloride (77mg,0.11mmol) were dissolved in tetrahydrofuran (6mL) and stirred at 60 degrees for 1 hour after displacing nitrogen 3 times. The reaction mixture was diluted with dichloromethane (50mL) and water (50mL), the aqueous phase was extracted with dichloromethane (50 mL. times.3), and the organic phase was washed once with saturated brine (100mL), then dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was chromatographed on silica gel column (eluent: 80% to 90% ethyl acetate/petroleum ether) to give N- [5- [2- (2-amino-5-methanesulfonyl-phenyl) ethynyl ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (6) as a yellow solid (140mg, 23% yield).

LCMS:t_R＝0.691min in 5-95AB_1.5min_220&254_Shimadzu.lcm chromatograph y(Merck RP18e 25-3mm),MS(ESI)m/z＝396.0[M+H]⁺.

Step 2.3 preparation of Compound EXP-4

N- [5- [2- (2-amino-5-methanesulfonyl-phenyl) ethynyl ] - [1,2,4] triazolo [1,5-a ] pyrid-2-yl ] cyclopropanecarboxamide (6) (110mg,0.278mmol) was dissolved in N, N-dimethylformamide (2mL), and potassium tert-butoxide (62mg,0.556mmol) was added and stirred at 90 degrees for 2 hours. The reaction mixture was diluted with ethyl acetate (30mL) and water (25mL), the aqueous phase was extracted with ethyl acetate (20mL), the organic phases were combined and washed once with brine (50mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was isolated and purified by basic preparation to give N- [5- (5-methanesulfonyl-1H-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (EXP-4) (2.4mg, 2% yield) as a white solid.

LCMS:t_R＝2.448min in 10-80AB_7min_220&254_Agilent.met,chromatography(Xbrige C18,30*2.1mm*3.5μm),MS(ESI)m/z＝396.1[M+H]⁺.

HPLC:t_R＝2.53min in 10-80CD_8min.met,chromatography(XBridge C18,2.1*50mm*5μm).

1H NMR(400MHz,DMSO-d₆):δ＝12.50(s,1H),11.28(s,1H),8.32(d,J＝1.6H z,1H),8.24(s,1H),7.90-7.80(m,2H),7.79-7.69(m,3H),3.23(s,3H),2.13-2.05(m,1H),0.92-0.86(m,4H).

Example 3: preparation of Compound EXP-5

The synthetic route is as follows:

step 3.1 preparation of Compound EXP-5

N- [5- (5-methanesulfonyl-1H-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (EXP-4) (30mg, 0.076mmol) was dissolved in N, N-dimethylformamide (0.5mL), and potassium carbonate (31mg,0.228mmol) and iodomethane (11mg,0.083mmol) were added and stirred at 10-12 degrees for 4 hours. The reaction mixture was extracted with ethyl acetate (20mL) and water (10mL), and the organic phase was washed once with brine (20mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product obtained was isolated and purified by basic preparation to give N- [5- (1-methyl-5-methanesulfonyl-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (EXP-5) (2.0mg, 6% yield) as a white solid.

LCMS:t_R＝2.246min in 10-80AB_7min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*30mm3um),MS(ESI)m/z＝410.0[M+H]⁺.

HPLC:t_R＝3.13min in 10-80AB_8min.met,chromatography(Ultimate C18 3.0*50mm3um).

1H NMR(400MHz,DMSO-d₆):δ＝11.13(s,1H),8.29(d,J＝1.2Hz,1H),7.91-7.83(m,2H),7.82-7.75(m,2H),7.39(dd,J＝7.2,0.8Hz,1H),7.15(s,1H),3.74(s,3H),3.20(s,3H),2.04-1.91(m,1H),0.90-0.68(m,4H).

Example 4: preparation of Compound EXP-6

The synthetic route is as follows:

step 4.1 preparation of Compound 8

N- [5- (5-Nitro-1H-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (70mg,0.193mmol) was dissolved in ethanol (1mL) and wet palladium on carbon (7mg) was added under a nitrogen atmosphere, and the reaction was allowed to displace hydrogen 3 times and stirred at 30 degrees (30psi) for 4 hours. The reaction mixture was filtered and the filtrate was spin dried to give crude product. The crude product was subjected to silica gel column chromatography (eluent: 5% methanol/dichloromethane) to give N- [5- (5-amino-1H-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (8) (25mg, yield 32%) as a yellow solid.

LCMS:t_R＝0.709min in 5-95AB_1.5min_220&254_Shimadzu.lcm chromatography(Merck RP-18e 25-3mm),MS(ESI)m/z＝333.0[M+H]⁺.

Step 4.2 preparation of Compound EXP-6

N- [5- (5-amino-1H-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (8) (25mg,0.075mmol) was dissolved in dichloromethane (1mL) and triethylamine (22mg,0.226mmol) and methanesulfonyl chloride (9mg,0.083mmol) were added and the mixture was stirred at 10-17 degrees for 1 hour. The reaction was quenched with saturated sodium bicarbonate (10mL) and extracted with dichloromethane (10mL × 2). The organic phases were combined and washed once with brine (10mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product thus obtained was isolated and purified by basic preparation to give N- [5- (5-methanesulfonamido-1H-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (EXP-6) (2mg, yield 6%) as a white solid.

LCMS:t_R＝2.353min in 10-80AB_7min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*30mm3um),MS(ESI)m/z＝411.0[M+H]⁺.

HPLC:t_R＝2.358min in 10-80CD_8min.met,chromatography(XBridge C18 2.1*50mm 5um).

1H NMR(400MHz,DMSO-d₆):δ＝11.96(s,1H),11.25(s,1H),9.31(s,1H),8.02(s,1H),7.79(d,J＝5.2Hz,2H),7.69-7.63(m,1H),7.55-7.44(m,2H),7.15(dd,J＝8.8,2.0Hz,1H),2.90(s,3H),2.12-1.95(m,1H),0.93-0.82(m,4H).

Example 5: preparation of Compound EXP-5

The synthetic route is as follows:

step 5.1 preparation of Compound 9

N- [ 5-acetylene- [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (5) (520mg,2.30mmol), 2-iodo-4-nitro-aniline (SM3) (606mg,2.30mmol), cuprous iodide (87mg,0.46mmol), triethylamine (1.16g,11.49mmol) and triphenylphosphine palladium dichloride (161mg,0.23mmol) were dissolved in tetrahydrofuran (6mL) and stirred at 60 ℃ for 1 hour after 3 replacements of nitrogen. Water (50mL) was added to the reaction mixture and extracted with dichloromethane (50mLx2), and the organic phase was washed once with saturated brine (50mL), dried over anhydrous sodium sulfate, and concentrated by filtration. The resulting crude product was chromatographed on silica gel (eluent: 70% ethyl acetate/petroleum ether) to give N- [5- [2- (2-amino-5-nitro-phenyl) acetylene ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (9) (340mg, 36% yield) as a yellow solid.

LCMS:t_R＝0.779min in 5-95AB_1.5min_220&254_Shimadzu.lcm chromatography(Merck RP18e 25-3mm),MS(ESI)m/z＝363.0[M+H]⁺.

Step 5.2 preparation of Compound 7

N- [5- [2- (2-amino-5-nitro-phenyl) acetylene ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (9) (340mg,0.94mmol) was dissolved in N, N-dimethylformamide (5mL) and potassium tert-butoxide (210mg,1.88mmol) was added and stirred at 90 degrees for 2 hours. The reaction mixture was added with water (10mL) and extracted with ethyl acetate (20mL), and the organic phase was washed twice with saturated brine (20mL), then dried over anhydrous sodium sulfate and concentrated by filtration. The resulting crude product was subjected to silica gel column chromatography (eluent: 80% ethyl acetate/petroleum ether) to give N- [5- (5-nitro-1H-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (24) (200mg, 58% yield) as a yellow solid.

1H NMR(400MHz,DMSO-d₆):δ＝12.65(s,1H),11.26(s,1H),8.72(d,J＝2.0Hz,2H),8.25(s,1H),8.13(dd,J＝9.2,2.4Hz,1H),7.89-7.72(m,3H),7.69(d,J＝9.2Hz,1H),2.17-2.02(m,1H),1.01-0.76(m,4H).

Step 5.3 preparation of Compound 10

N- [5- (5-Nitro-1H-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (7) (190mg,0.524mmol) was dissolved in N, N-dimethylformamide (2mL), potassium carbonate (217mg,1.57mmol) and iodomethane (81mg,0.577mmol) were added and stirred at 30 degrees for 2 hours. The reaction mixture was added with water (10mL) and extracted with ethyl acetate (20mL), and the organic phase was washed once with saturated brine (20mL), then dried over anhydrous sodium sulfate and concentrated by filtration. The resulting crude product was subjected to silica gel column chromatography (eluent: 80% ethyl acetate/petroleum ether) to give N- [5- (1-methyl-5-nitro-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (10) (90mg, 32% yield) as a yellow solid.

LCMS:t_R＝0.753min in 5-95AB_1.5min_220&254_Shimadzu.lcm chromatography(Merck RP18e 25-3mm),MS(ESI)m/z＝377.0[M+H]⁺.

1H NMR(400MHz,DMSO-d₆):δ＝11.13(s,1H),8.71(d,J＝2.4Hz,1H),8.17(dd,J＝9.2,2.0Hz,1H),7.93-7.75(m,3H),7.41(d,J＝7.2Hz,1H),7.21(s,1H),3.75(s,3H),2.22-1.95(m,1H),0.83-0.60(m,4H).

Step 5.4 preparation of Compound 11

N- [5- (1-methyl-5-nitro-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (10) (80mg,0.212mmol) was dissolved in ethanol (1mL) and palladium on carbon (10mg, palladium purity 10 wt%) was added and hydrogen was replaced 3 times under an atmosphere of nitrogen, and the reaction was stirred under an atmosphere of hydrogen (30Psi) at 30 ℃ for 4 hours. The reaction solution was filtered, and the filtrate was spin-dried to give a crude product, which was subjected to silica gel column chromatography (eluent: 4% methanol/dichloromethane) to give N- [5- (1-methyl-5-amino-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyrid-2-yl ] cyclopropanecarboxamide (11) (30mg, 34% yield) as a yellow solid.

LCMS:t_R＝0.563min in 5-95AB_1.5min_220&254_Shimadzu.lcm chromatography(Merck RP18e 25-3mm),MS(ESI)m/z＝347.1[M+H]⁺.

Step 5.5 preparation of Compound EXP-7

N- [5- (1-methyl-5-amino-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (11) (30mg,0.087mmol) was dissolved in dichloromethane (1mL) and triethylamine (26mg,0.26mmol) and methanesulfonyl chloride (10mg,0.095mmol) were added and stirred at 10-17 degrees for 16 hours. The reaction was quenched with saturated sodium bicarbonate (10mL) and extracted with dichloromethane (10mL × 2). The organic phases were combined and washed once with brine (10mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product obtained was isolated and purified by basic preparation to give N- [5- (5-methanesulfonyl-1-methyl-indol-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (EXP-7) (2.3mg, 6% yield) as a white solid.

LCMS:t_R＝2.231min in 10-80AB_7min_220&254_Agilent.met,chromatography(Xtimate C18 2.1*30mm3um),MS(ESI)m/z＝447.0[M+Na]⁺.

HPLC:t_R＝3.10min in 10-80CD_8min.met,chromatography(Ultimate C18 3.0*50mm3um).

1H NMR(400MHz,DMSO-d₆):δ＝11.12(s,1H),9.32(s,1H),7.87-7.73(m,2H),7.63-7.50(m,2H),7.34(dd,J＝7.2,1.6Hz,1H),7.21(dd,J＝8.8,2.0Hz,1H),6.91(s,1H),3.66(s,3H),2.89(s,3H),2.05-1.90(m,1H),0.89-0.74(m,4H).

Example 6: preparation of Compound EXP-8

The synthetic route is as follows:

step 6.1 preparation of Compound 12

5-methylbenzothiophene (SM4) (200mg,1.35mmol) was dissolved in tetrahydrofuran (4mL), and n-butyllithium (2.5M n-hexane solution, 0.81mL) was slowly added dropwise thereto at-75 ℃ under a nitrogen atmosphere. After stirring for 60 min, triisopropyl borate (381mg,2.03mmol) was added, then slowly raised to 15-20 ℃ and stirring continued for 16 h. The reaction was quenched with 1M hydrochloric acid solution (20mL) and the aqueous phase was extracted with ethyl acetate (20mL x 3), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was spun dry to give the desired product [ 5-methylbenzothiophen-2-yl ] boronic acid (29) (300mg, crude, white solid).

1H NMR(400MHz,DMSO):δ＝8.44(s,2H),7.87(s,1H),7.83(d,J＝8.4Hz,1H),7.68(s,1H),7.20(d,J＝8.4Hz,1H),2.41(s,3H).

Step 6.2 preparation of Compound EXP-8

[ 5-Methylbenzothiophen-2-yl ] boronic acid (12) (150mg,0.78mmol) and (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-amine) cyclopropanecarboxamide (Int-1) (220mg,0.78mmol) were dissolved in dioxane (5mL) and water (1mL), and potassium carbonate (324mg,2.34mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (114mg,0.156mmol) were added rapidly under a nitrogen atmosphere. The mixture was replaced with nitrogen three times, and then stirred at 90 ℃ for 2 hours. The reaction mixture was filtered, the filtrate was diluted with ethyl acetate (20mL) and water (20mL), the aqueous phase was extracted with ethyl acetate (20 mL. times.3), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, spin-dried and chromatographed on silica gel (eluent: 20% -100% ethyl acetate/petroleum ether) to give crude product (180mg, 70% purity). 100mg of crude product were then purified by HPLC (separation column: A gela ASB 150 x 25mm x 5 um; mobile phase: A: water (0.05% HCl) and B: acetonitrile; gradient: B50-80%) and the yellow target product N- [5- [ 5-methylbenzothien-2-yl ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] cyclopropanecarboxamide (EXP-8) (14.3mg, yield 4.64%, purity 97.53%, HCl salt) was obtained, the remaining 80mg of crude product was used for further reactions.

LCMS:t_R＝3.758min in 10-80CD_7min_220&254_Agilent.met,chromatography(Xbrige C18 30*2.1mm 3.5um),MS(ESI)m/z＝349.2[M+H]⁺.

HPLC:t_R＝3.61min in 10-80CD_8min.met,chromatography(XBridge C18 2.1*50mm*5um).

1H NMR(400MHz,DMSO-d₆):δ＝11.24(s,1H),8.77(s,1H),7.97(d,J＝8.4Hz,1H),7.81-7.70(m,4H),7.32(d,J＝8.4Hz,2H),2.46(s,3H),2.20-2.01(m,1H),0.92-0.82(m,4H).

Example 7: preparation of Compound EXP-9

The synthetic route is as follows:

step 7.1 preparation of Compound 13

[ 5-methylbenzothiophen-2-yl ] boronic acid (12) (170mg,0.89mmol) was dissolved in tetrahydrofuran (2mL), and pinacol (104mg,0.89mmol) and trifluoroacetic acid (20mg,0.18mmol) were added thereto. The reaction solution was substituted with nitrogen three times and reacted at 15 to 20 ℃ for 2 hours. The reaction mixture was diluted with ethyl acetate (20mL) and water (20mL) and the aqueous phase was extracted with ethyl acetate (20mL × 3), the organic phase was washed with brine (20mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated and dried to give 4,4,5, 5-tetramethyl-2- (5-methylbenzothien-2-yl) -1,3, 2-dioxaborane (13) (200mg, yield 82%) as a white target product, which was used directly in the next reaction without further purification.

1H NMR(400MHz,DMSO-d₆):δ＝7.89(d,J＝8.4Hz,1H),7.82(s,1H),7.74(s,1H),7.27(d,J＝8.4Hz,1H),2.42(s,3H),1.32(s,12H).

Step 7.2 preparation of Compound 14

4,4,5, 5-tetramethyl-2- (5-methylbenzothien-2-yl) -1,3, 2-dioxaborane (13) (200mg,0.73mm ol) was dissolved in carbon tetrachloride (5mL), N-bromosuccinimide (130mg,0.73mmol) and benzoyl peroxide (18mg,0.07mmol) were added thereto and stirring was continued at 80 ℃ for 2 hours. The reaction mixture was diluted with ethyl acetate (20mL) and water (20mL) and the aqueous phase was extracted with ethyl acetate (20mL x 3), the organic phase was washed with brine (20mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated and dried to give the white desired product 2- [5- (bromomethyl) -benzothien-2-yl ] -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane (14) (250mg, 97% yield) which was used directly in the next reaction without further purification.

1H NMR(400MHz,DMSO-d₆):δ＝8.05-8.01(m,2H),7.92(s,1H),7.54-7.46(m,1H),4.86(s,2H),1.33(s,12H).

Step 7.3 preparation of Compound Int-2

2- [5- (bromomethyl) -benzothien-2-yl ] -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane (14) (250mg,0.71mmol) and potassium carbonate (196mg,1.42mmol) were dissolved in acetonitrile (4mL), to which 1, 1-dioxothiomorpholine (96mg,0.71mmol) was added and stirring was continued at 15-20 ℃ for 2 hours. The reaction solution was filtered and spin dried to give 4- [ [2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2 yl) benzothien-5-yl ] methyl ] -1, 1-dioxothiomorpholine (Int-2) (200mg, crude) as a white solid, which was used directly in the next reaction without further purification.

1H NMR(400MHz,DMSO-d₆):δ＝7.98(d,J＝8.4Hz,1H),7.90-7.89(m,2H),7.42(d,J＝8.4Hz,1H),3.79(s,2H),3.15-3.08(m,4H),2.93-2.84(m,4H),1.32(s,12H).

Step 7.4 preparation of Compound EXP-9

4- [ [2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2 yl) benzothien-5-yl]Methyl radical]-1, 1-Dioxothiomorpholine (Int-2) (200mg,0.49mmol) and (5-bromo- [1,2, 4)]Triazole [1,5-a ]]Pyridin-2-amine) Cyclopropanecarboxamide (Int-1) (110mg,0.39mmol) was dissolved in dioxane (3mL) and water (0.6mL), and potassium carbonate (204mg,1.47mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (72mg,0.098mmol) were added rapidly under nitrogen. The mixture was replaced with nitrogen three times, and then stirred at 90 ℃ for 2 hours. The reaction mixture was filtered, the filtrate was diluted with ethyl acetate (20mL) and water (20mL), the aqueous phase was extracted with ethyl acetate (20 mL. times.3), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, spin-dried and chromatographed on silica gel (eluent: 20% -100% ethyl acetate/petroleum ether) to give crude product (180mg, 70% purity). 100mg of crude product were then purified by HPLC (separation column: AgelaDuraShell C18150 mm 25mm 5 um; mobile phase: A: water (0.04% NH)₃.H₂O+10mM NH₄HCO₃) And B is acetonitrile; gradient: b35-65%) and obtaining the white target product N- [5- [5- [ (1, 1-dioxygen thiomorpholin-4-yl) methyl)]Benzothiophen-2-yl]-[1,2,4]Triazole [1,5-a ]]Pyridin-2-yl]Cyclopropanecarboxamide (EXP-9) (28.6mg, yield 12%, purity 99.14%).

LCMS:t_R＝3.420min in 10-80AB_7min_220&254_Shimadzu.lcm(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝482.3[M+H]⁺.

HPLC:t_R＝3.66min in 10-80AB_8min.met,chromatography.

1H NMR(400MHz,DMSO-d₆):δ＝11.23(s,1H),8.83(s,1H),8.06(d,J＝8.4Hz,1H),7.93(s,1H),7.79-7.70(m,3H),7.47(d,J＝8.4Hz,1H),3.83(s,2H),3.17-3.04(m,4H),3.01-2.89(m,4H),2.14-2.08(m,1H),0.97-0.78(m,4H).

The example compounds of the following table 2 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 2 ]

Example 8: preparation of Compound EXP-14

The synthetic route is as follows:

step 8.1 preparation of Compound 47

6-bromo-benzothiophene (SM6) (100mg,0.47mmol) and methanesulfonamide (SM7) (67mg,0.70mmol) were dissolved in N, N-dimethylacetamide (2mL), and copper iodide (9mg,0.47mmol) and potassium carbonate (130mg,0.94mmol) were added thereto at room temperature, followed by microwave reaction at 190 ℃ for 2 hours. The reaction mixture was filtered, the filtrate was diluted with ethyl acetate (10mL) and water (10mL) and the layers were separated, the aqueous phase was extracted with ethyl acetate (10 mL. times.3), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and spin-dried, and column chromatography on silica gel (eluent: 0% to 50% ethyl acetate/petroleum ether) was used to give the desired product N- (benzothien-6-yl) methanesulfonamide (47) (70mg, 66% yield) as a light brown color.

1H NMR(400MHz,DMSO-d₆):δ＝9.83(s,1H),7.85-7.79(m,2H),7.67(d,J＝5.2Hz,1H),7.40(d,J＝5.2Hz,1H),7.25(dd,J＝8.4,2.0Hz,1H),3.00(s,3H).

Step 8.2 preparation of Compound 48

N- (benzothien-6-yl) methanesulfonamide (47) (70mg,0.31mmol) was dissolved in tetrahydrofuran (2mL), to which N-butyllithium (2.5M,0.62mL) was slowly added dropwise under a nitrogen atmosphere at-75 ℃. After stirring for 60 minutes, triisopropyl borate (174mg,0.92mmol) was added, then slowly raised to 15-20 ℃ and stirring continued for 16 hours. The reaction was quenched with 1M hydrochloric acid solution (10mL) and the aqueous phase was extracted with ethyl acetate (10mL × 3), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was spun dry to give [6- (methanesulfonamide) benzothien-2-yl ] boronic acid (48) (70mg, crude) as a pale blue solid.

1H NMR(400MHz,DMSO-d₆):δ＝9.88(s,1H),8.46(s,2H),7.89(s,1H),7.84(d,J＝8.8Hz,1H),7.75(s,1H),7.23(dd,J＝8.8,1.6Hz,1H),3.01(s,3H).

Step 8.3 preparation of Compound EXP-14

[6- (Methanesulfonamide) benzothien-2-yl group]Boronic acid (48) (70mg,0.26mmol) and (5-bromo- [1,2, 4)]Triazole [1,5-a ]]Pyridin-2-amine) Cyclopropanecarboxamide (Int-1) (58mg,0.21mmol) was dissolved in dioxane (1mL) and water (0.2mL), and potassium carbonate (107mg,0.77mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (19mg,0.026mmol) were added rapidly under nitrogen. The mixture was replaced with nitrogen three times, and then stirred at 90 ℃ for 2 hours. The reaction mixture was filtered, the filtrate was diluted with ethyl acetate (20mL) and water (20mL), the aqueous phase was extracted with ethyl acetate (20 mL. times.3), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and dried before purification by HPLC (separation column: AgelaDuraShell C18250. times.25 mm. times.10 um; mobile phase: A: water (0.04% NH)₃.H₂O+10mM NH₄HCO₃) And B is acetonitrile; gradient: b40-70%) and obtaining the white target product N- [5- [6- (methane sulfonamide) benzothiophen-2-yl]-[1,2,4]Triazole [1,5-a ]]Pyridin-2-yl]Cyclopropanecarboxamide (EXP-14) (10.6mg, yield 9%, purity 93.21%).

LCMS:t_R＝3.880min in 10-80AB_7min_220&254_Shimadzu.lcm(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝428.2[M+H]⁺.

HPLC:t_R＝4.79min in 10-80AB_8min.met,chromatography.

1H NMR(400MHz,DMSO-d₆):δ＝11.21(s,1H),10.06(s,1H),8.78(s,1H),7.94(d,J＝8.8Hz,1H),7.86(s,1H),7.79-7.67(m,3H),7.32(dd,J＝8.8,1.6Hz,1H),3.08(s,3H),2.17-2.01(m,1H),0.97-0.75(m,4H).

Example 9: preparation of Compound EXP-15

The synthetic route is as follows:

step 9.1 preparation of Compound 50

To a solution of (benzothien-5-yl) -carbamic acid tert-butyl ester (49) (371mg,1.49mmol) in anhydrous tetrahydrofuran (7.5mL) was added sodium hydride (72mg, 60% pure in mineral oil, 1.80mmol) portionwise under ice-water bath conditions at 0 ℃. After stirring for 30min, iodomethane (0.2mL, d 2.28g/mL,3.21mmol) was added. The reaction was gradually warmed to room temperature and stirring was continued for 17 hours. The resulting pale yellow turbidity was quenched with 20% ammonium chloride solution (10mL) and extracted with tert-butyl methyl ether (12mL,10mL,8 mL). The organic phases were combined and washed with water (12 mL. times.2) and saturated brine (15mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give (benzothien-5-yl) -carbamic acid tert-butyl ester (50) (366mg,1.39mmol, 93% yield, light yellow solid) which was used directly in the next reaction.

¹H NMR(400MHz,CDCl₃):δ＝7.80(d,J＝8.4Hz,1H),7.65(s,1H),7.46(d,J＝5.2Hz,1H),7.29(d,J＝5.6Hz,1H),7.23(dd,J＝8.8,1.6Hz,1H),3.31(s,3H),1.45(s,9H).

Step 9.2 preparation of Compound 15

To a solution of tert-butyl (benzothien-5-yl) -methylcarbamate (50) (366mg,1.39mmol) in anhydrous tetrahydrofuran (9.0mL) at-60 ℃ under a nitrogen atmosphere was added dropwise n-butyllithium (0.8mL,2.5M in hexane) over five minutes. After the reaction mixture was kept at-60 ℃ and stirred for 50 minutes, triisopropyl borate (0.4mL, d 0.818g/mL) was added dropwise over five minutes, followed by natural warming to room temperature and stirring was continued for 17 hours. The resulting yellow turbid solution was quenched with 0.2M dilute hydrochloric acid (10mL) and extracted with ethyl acetate (6 mL. times.3). The combined organic phases were washed with brine (10mL), dried over anhydrous sodium sulfate, filtered and concentrated to dryness to give 5- (tert-butoxycarbonyl) methylaminobenzothiophene-2-boronic acid (15) (443mg, about 90% purity, 93% yield, light yellow solid) which was used directly in the next reaction.

¹H NMR(400MHz,DMSO-d₆):δ＝8.50(brs,2H),7.94-7.87(m,2H),7.77(d,J＝1.0Hz,1H),7.28(dd,J＝8.8,2.0Hz,1H),3.23(s,3H),1.38(s,9H).

Step 9.3 preparation of Compound 16

To a mixed solution of N- (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -cyclopropanecarboxamide (Int-1) (244mg,0.868mmol) and 5- (tert-butoxycarbonyl) methylaminobenzothiophene-2-boronic acid (15) (443mg, 90% purity, 1.30mmol) in 1, 4-dioxane (7.0mL) and water (1.5mL) was added 1, 1' -bis (diphenylphosphino) ferrocene dichloropalladium (40mg,0.055mmol) and potassium phosphate (503mg,2.37 mmol). The mixture was replaced with nitrogen several times, and then heated to 95 ℃ under nitrogen atmosphere and stirred continuously for 20 hours. The resulting yellow suspension was cooled and diluted with water (15 mL). The resulting precipitate was collected by filtration and washed with water (5 mL. times.2) and dried in vacuo to give {5- [5- (tert-butoxycarbonyl) methylaminobenzothien-2-yl ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (16) (495mg, about 80% purity, 98% yield, light brown solid) which was used directly in the next reaction.

LCMS:t_R＝0.998min in 5-95AB_1.5min_220&254_Shimadzu.lcm,MS(ESI)m/z＝464.1[M+H]⁺.

¹H NMR(400MHz,DMSO-d₆):δ＝11.21(brs,1H),8.76(s,1H),8.04(d,J＝8.8Hz,1H),7.86(s,1H),7.81-7.68(m,3H),7.45-7.37(m,1H),3.27(s,3H),2.17-2.04(m,1H),1.41(s,9H),1.40-1.29(m,4H).

Step 9.4 preparation of Compound 17

{5- [5- (tert-butoxycarbonyl) methylaminobenzothien-2-yl ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (16) (495mg, 80% purity, 0.855mmol) was suspended in anhydrous dichloromethane (5.6mL) at 10 ℃ and trifluoroacetic acid (2.8mL, d ═ 1.54g/mL,37.82mmol) was added dropwise thereto. After stirring the resulting dark brown solution at room temperature for 2 hours, most of the volatiles were removed by bubbling a stream of nitrogen and treated with 1M potassium carbonate solution (20 mL). The resulting mixture was extracted with dichloromethane/methanol (30mL × 3, v/v ═ 5/1) with mixing. The combined extracts were washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, concentrated and evaporated to dryness to give {5- (5-methylaminobenzothien-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (17) (234mg crude) which was used directly in the next reaction.

LCMS:t_R＝0.783min in 5-95AB_1.5min_220&254_Shimadzu.lcm,MS(ESI)m/z＝364.1[M+H]⁺.

Step 9.5 preparation of Compound EXP-15

To a suspension of {5- (5-methylaminobenzothien-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (17) (117mg) in anhydrous dichloromethane (4mL) was added triethylamine (0.1mL, d ═ 0.727g/mL) and methanesulfonic anhydride (86mg) at 10 ℃. The reaction mixture was stirred at 10 ℃ for 20 hours, and the resulting yellow-brown suspension was concentrated under reduced pressure to remove most of the volatiles and diluted with water. The resulting precipitate was collected by filtration and washed with water (2 mL. times.2), preparative separated by high performance liquid chromatography (50-80% acetonitrile/0.04% ammonia +10mM ammonium bicarbonate), the pure fractions collected combined, concentrated under reduced pressure and lyophilized to give the final product {5- (5-methanesulfonylmethylaminobenzothien-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (EXP-15) (13.1mg, 100% purity, 7% two step yield, white solid).

LCMS:t_R＝2.907min in 10-80AB_4min_220&254_Shimadzu.lcm,MS(ESI)m/z＝442.2[M+H]⁺.

t_R＝2.975min in 10-80AB_4min_220&254_Shimadzu.lcm,MS(ESI)m/z＝442.2[M+H]⁺.

HPLC:t_R＝2.64min in 10-80CD_8min.met(XBridge Shield RP18 5um).

¹H NMR(400MHz,DMSO-d₆):δ＝11.24(brs,1H),8.83(s,1H),8.34-7.96(m,2H),7.95-7.36(m,4H),3.45(s,3H),3.00(s,3H),2.16-1.93(m,1H),1.10-0.60(m,4H).

The example compounds of the following table 3 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 3 ]

Example 10: preparation of Compound EXP-25

The synthetic route is as follows:

step 10.1 preparation of Compound EXP-25

4- [ [2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2 yl) benzothien-5-yl]Methyl radical]Morpholine (Int-3) (100mg,0.28mmol) and (5-bromo- [1,2, 4)]Triazole [1,5-a ]]Pyridin-2-amine) Cyclopropanecarboxamide (Int-1) (39mg,0.14mmol) was dissolved in dioxane (0.5mL) and water (0.1mL), and potassium carbonate (115mg,0.83mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (20mg,0.027mmol) were added rapidly under a nitrogen atmosphere. The mixture was replaced with nitrogen three times, and then stirred at 90 ℃ for 2 hours. The reaction mixture and EB5-42 were combined and filtered, the filtrate was diluted with ethyl acetate (20mL) and water (20mL), the aqueous phase was extracted with ethyl acetate (20 mL. times.3), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, spun dry and purified by preparative HPLC (separation column: AgelaDuraShell C18250 mm 25mm 10 um; mobile phase: A: water (0.04% NH)₃.H₂O+10mM NH₄HCO₃) And B is acetonitrile(ii) a Gradient: b45-75%) to obtain white target product N- [5- [5- (morpholine) benzothiophen-2-yl]-[1,2,4]Triazole [1,5-a ]]Pyridin-2-yl]Cyclopropanecarboxamide (EXP-25) (15mg, yield 10%, purity 100%).

HPLC:t_R＝2.75min in 10-80CD_8min.met,chromatography(XBridge Shield RP18 5um).

1H NMR(400MHz,DMSO-d₆):δ＝11.22(s,1H),8.82(s,1H),8.04(d,J＝8.4Hz,1H),7.89(s,1H),7.80-7.68(m,3H),7.45(d,J＝8.4Hz,1H),3.66-3.54(m,6H),2.43-2.37(m,4H),2.14-2.02(m,1H),0.91-0.81(m,4H).

The example compounds of the following table 4 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 4]

Example 11: preparation of Compound EXP-27

The synthetic route is as follows:

step 11.1 preparation of Compound Int-4

2- [5- (bromomethyl) -benzothien-2-yl ] -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane (14) (100mg,0.28mmol) and potassium carbonate (78mg,0.57mmol) were dissolved in acetonitrile (2mL), 4-difluoropiperidine (SM8) (34mg,0.28mmol) was added thereto and stirring was continued at 15-20 ℃ for 2 hours. The reaction mixture was diluted with ethyl acetate (20mL) and water (20mL) and the aqueous phase was extracted with ethyl acetate (20mL × 3), the organic phase was dried over anhydrous sodium sulfate and filtered, concentrated and dried to give 4, 4-difluoro-1- [ [2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2 yl) benzothien-5-yl ] methyl ] piperidine (Int-4) (150mg, crude) as a brown target product which was used in the following reaction without further purification.

Step 11.2 preparation of Compound EXP-27

4, 4-difluoro-1- [ [2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2 yl) benzothien-5-yl]Methyl radical]Piperidine (Int-4) (100mg,0.25mmol) and (5-bromo- [1,2, 4)]Triazole [1,5-a ]]Pyridin-2-amine) Cyclopropanecarboxamide (Int-1) (36mg,0.13mmol) was dissolved in dioxane (2mL) and water (0.4mL), and potassium carbonate (105mg,0.76mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (19mg,0.025mmol) were added rapidly under nitrogen. The mixture was replaced with nitrogen three times, and then stirred at 90 ℃ for 2 hours. The reaction mixture was filtered, the filtrate was diluted with ethyl acetate (20mL) and water (20mL), the aqueous phase was extracted with ethyl acetate (20 mL. times.3), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and dried before purification by preparative HPLC (separation column: AgelaDuraShell C18250. times.25 mm. times.10 um; mobile phase: A: water (0.04% NH)_3.H₂O+10mM NH₄HCO₃) And B is acetonitrile; gradient: b55-85%) and obtaining light yellow target product N- [5- [5- (4, 4-difluoro-1-piperidine) methyl]Benzothiophen-2-yl]-[1,2,4]Triazole [1,5-a ]]Pyridin-2-yl]Cyclopropanecarboxamide (EXP-27) (20mg, yield 13%, purity 95.12%).

LCMS:t_R＝3.324min in 10-80AB_7min_220&254_Shimadzu.lcm(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝468.3[M+H]⁺.

HPLC:t_R＝3.36min in 10-80CD_8min.met,chromatography(XBridge Shield RP18 5um).

1H NMR(400MHz,DMSO-d₆):δ＝11.22(s,1H),8.82(s,1H),8.04(d,J＝8.0Hz,1H),7.89(s,1H),7.76-7.74(m,3H),7.45(d,J＝8.4Hz,1H),3.70(s,2H),3.31-3.27(m,4H),2.17-2.06(m,1H),2.04-1.87(m,4H),0.98-0.83(m,4H).

Example 12: preparation of Compounds EXP-28, EXP-29, EXP-30, EXP-31

The synthetic route is as follows:

step 12.1 preparation of Compound 18

To a solution of 5-bromobenzothiophene (SM9) (8.61g,39.6mmol) and tert-butyl carbamate (6.45g,55.04mmol) in 1, 4-dioxane (180mL) was added cesium carbonate (25.67g,78.80mmol) and palladium acetate (569mg,2.53mmol), and 4, 5-bis diphenylphosphine-9, 9-dimethylxanthene (2.66g,4.59mmol) under a nitrogen stream. The mixture was replaced with nitrogen several times, and then heated to 100 ℃ under nitrogen atmosphere and stirred continuously for 20 hours. The resulting dark brown suspension was cooled, diluted with ethyl acetate (200mL) and the insoluble material was removed by filtration. The filtrate was washed with water (200mL) and saturated brine (200mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by automatic preparative chromatography (eluent: 1-5% ethyl acetate/petroleum ether), the combined pure fractions were collected and concentrated to dryness to give (benzothien-5-yl) -carbamic acid tert-butyl ester (18) (5.58g,22.38mmol, 57% yield, off-white solid).

¹H NMR(400MHz,CDCl₃):δ＝8.00(s,1H),7.75(d,J＝8.8Hz,1H),7.43(d,J＝5.6Hz,1H),7.26(d,J＝5.6Hz,1H),7.20(dd,J＝8.4,2.0Hz,1H),6.58(brs,1H),1.54(s,9H).

Step 12.2 preparation of Compound 19

To a solution of (benzothien-5-yl) -carbamic acid tert-butyl ester (18) (3.00g,12.03mmol) in anhydrous tetrahydrofuran (55mL) at-60 ℃ under a nitrogen atmosphere was added n-butyllithium (14.4mL,2.5M in hexane) dropwise continuously for 30 minutes. After the reaction mixture was kept at-60 ℃ and stirred for 60 minutes, triisopropyl borate (4.6mL, d 0.818g/mL) was added dropwise continuously for 30 minutes, followed by natural warming to room temperature and stirring was continued for 18 hours. The resulting yellow turbid solution was quenched with 0.4M dilute hydrochloric acid (90mL) and extracted with ethyl acetate (60 mL. times.3). The combined organic phases were washed with brine (100mL), dried over anhydrous sodium sulfate, filtered and concentrated to dryness to afford 5- (tert-butoxycarbonyl) aminobenzothiophene-2-boronic acid (19) (4.07g, about 80% purity, 92% yield, off-white solid) which was used directly in the next reaction.

¹H NMR(400MHz,DMSO-d₆):δ＝9.42(brs,1H),8.44(d,J＝1.6Hz,1H),8.05(brs,1H),7.87-7.79(m,2H),7.42-7.35(m,1H),7.34(s,1H),1.49(s,9H).

Step 12.3 preparation of Compound 20

To a mixed solution of N- (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -cyclopropanecarboxamide (Int-1) (967mg,3.44mmol) and 5- (tert-butoxycarbonyl) aminobenzothiophene-2-boronic acid (19) (2.16g, about 80% purity, 5.88mmol) in 1, 4-dioxane (27.5mL) and water (5.5mL) was added 1, 1' -bis (diphenylphosphino) ferrocene dichloropalladium (151mg,0.206mmol) and potassium phosphate (1.97g,9.29mmol) under a nitrogen stream. The mixture was replaced with nitrogen several times, and then heated to 95 ℃ under nitrogen atmosphere and stirred continuously for 20 hours. The resulting dark brown suspension (containing the pilot batch EB3-19) was cooled and diluted with water (55 mL). The resulting precipitate was collected by filtration and washed with water (10 mL. times.2) and dried in vacuo to afford {5- [5- (tert-butoxycarbonyl) aminobenzothiophen-2-yl ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (20) (2.38g, about 70% pure crude, brown solid) which was used directly in the next reaction.

LCMS:t_R＝0.975,1.111min in 5-95AB_1.5min_220&254.lcm,MS(ESI)m/z＝450.1[M+H]⁺.

Step 12.4 preparation of Compound EXP-28

{5- [5- (tert-butoxycarbonyl) aminobenzothiophen-2-yl ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (20) (2.38g, about 70% pure crude) was suspended in anhydrous dichloromethane (20mL) at 10 ℃ and trifluoroacetic acid (10mL, d ═ 1.54g/mL) was added dropwise thereto. After stirring the resulting solution at 10 ℃ for 2 hours, most of the volatiles were removed by bubbling a stream of nitrogen and treated with 1M potassium carbonate solution (50 mL). The resulting precipitate was collected by filtration and slurried with methylene chloride/methanol (10mL, 1:1 by volume). The resulting solid was dried in vacuo to give {5- (5-aminobenzothiophen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (EXP-28) (1.60g crude, about 70% purity, 93% two-step yield, yellow solid) which was used in the next reaction without purification.

LCMS:t_R＝0.780min in 5-95AB_1.5min_220&254_Shimadzu.lcm,MS(ESI)m/z＝350.0[M+H]⁺.

¹H NMR(400MHz,DMSO-d₆):δ＝11.19(brs,1H),8.63(s,1H),7.80-7.62(m,4H),7.60-7.45(m,2H),5.24(brs,2H),2.23-1.97(m,1H),1.38-1.17(m,4H).

Step 12.5 preparation of Compound EXP-29

Acetic anhydride (30 μ L, d ═ 1.09g/mL) was added to a suspension of {5- (5-aminobenzothiophen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (EXP-28) (100mg crude, about 70% purity) and triethylamine (0.1mL, d ═ 0.727g/mL) in dry dichloromethane (4mL) at 15 ℃. The reaction mixture was stirred at 15 ℃ for 20 hours, and the resulting yellow-brown suspension was concentrated under reduced pressure to remove most of the volatiles and diluted with water (8 mL). The resulting precipitate was collected by filtration and washed with water (2mL × 2) and preparative separated by high performance liquid chromatography (30-60% acetonitrile/0.04% ammonia +10mM ammonium bicarbonate), the pure fractions were collected, concentrated under reduced pressure and lyophilized to give the final product {5- (5-acetamidobenzothiophen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (EXP-29) (11.2mg, 100% purity, 14% yield, white solid).

LCMS:t_R＝0.877min in 5-95 AB_1.5min_220&254_Shimadzu.lcm,MS(ESI)m/z＝392.1[M+H]⁺.

t_R＝3.715min in 10-80 AB_7min_220&254_Shimadzu.lcm,MS(ESI)m/z＝392.2[M+H]⁺.

HPLC:t_R＝2.52min in 10-80 CD_8min.met(XBridge Shield RP18 5um).

¹H NMR(400MHz,DMSO-d₆):δ＝11.24(brs,1H),10.15(brs,1H),8.83(s,1H),8.41(s,1H),7.99(d,J＝7.2Hz,1H),7.84-7.66(m,3H),7.52(d,J＝7.2Hz,1H),2.26-2.16(m,1H),2.10(s,3H),1.01-0.72(m,4H).

Step 12.6 preparation of Compound EXP-14

To a solution of {5- (5-aminobenzothiophen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (EXP-28) (100mg crude, about 70% purity) in anhydrous N, N-dimethylformamide (1.8mL) was added sodium hydride (27mg,60 wt% in mineral oil) at 0 ℃. After stirring at 0 ℃ for 30 minutes, 2-methoxyethanesulfonyl chloride (SM22) (57mg,0.349mmol) was added dropwise. After the reaction solution was warmed to 15 ℃ and continuously stirred for 14 hours, it was quenched with 1M aqueous ammonium chloride (10mL), followed by extraction with ethyl acetate (12mL,10mL and 8 mL). The organic phases were combined, washed successively with water (10 mL. times.2) and saturated brine (15mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was isolated using thin layer prep plates (developer: dichloromethane/ethyl acetate 1/3) to give an orange oil, followed by preparative separation by high performance liquid chromatography (35-65% acetonitrile/0.04% ammonia +10mM ammonium bicarbonate), the combined pure fractions were collected, concentrated under reduced pressure and lyophilized to give the final product {5- [5- (2-methoxy) ethanesulfonylamidobenzothien-2-yl ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl } cyclopropanecarboxamide (EXP-30) (2.9mg, 98.09% purity, 3% yield, off-white solid).

LCMS:t_R＝1.340min in 5-95 AB_2min_220&254_Shimadzu.lcm,MS(ESI)m/z＝472.2[M+H]⁺.

t_R＝5.648min in 10-80 AB_7min_220&254_Shimadzu.lcm,MS(ESI)m/z＝472.1[M+H]⁺.

HPLC:t_R＝3.33min in 10-80 AB_8min.met.

¹H NMR(400MHz,DMSO-d₆):δ＝11.24(brs,1H),8.80(s,1H),8.02(d,J＝8.8Hz,1H),7.86-7.65(m,4H),7.31(dd,J＝8.4,2.0Hz,1H),3.68(t,J＝6.0Hz,2H),3.46(t,J＝6.0Hz,2H),3.19(s,3H),2.23-2.01(m,1H),0.98-0.78(m,4H).

Step 12.7 preparation of Compound EXP-31

Cis-N- (5-bromo- [1,2, 4)]Triazole [1,5-a ]]Pyridin-2-yl) -2-fluoro-cyclopropanecarboxamide (Int-2) (97mg,0.32mmol) and [5- (methanesulfonamide) benzothien-2-yl]Boric acid (20) (110mg,0.41mmol) was dissolved in dioxane (2mL) and water (0.4mL), and potassium carbonate (168mg,1.22mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (30mg,0.041mmol) were added rapidly under a nitrogen atmosphere. The mixture was replaced with nitrogen three times, and then stirred at 90 ℃ for 2 hours. Reaction ofThe mixture was filtered, the filtrate diluted with ethyl acetate (20mL) and water (20mL), the aqueous phase extracted with ethyl acetate (20 mL. times.3), the organic phase dried over anhydrous sodium sulfate, filtered, concentrated and dried before purification by HPLC (separation column: AgelaDuraShell C18250: 25 mm. times.10 um; mobile phase: A: water (0.04% NH)₃.H₂O+10mM NH₄HCO₃) And B is acetonitrile; gradient: b35-65%) and obtaining the white target product 2-fluoro-N- [5- [5- (methane sulfonamide) benzothiophen-2-yl]-[1,2,4]Triazole [1,5-a ]]Pyridin-2-yl]Cyclopropanecarboxamide (EXP-31) (12.9mg, yield 7%, purity 96.66%).

LCMS:t_R＝3.732min in 10-80AB_7min_220&254_Shimadzu.lcm(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝446.2[M+H]⁺.

HPLC:t_R＝1.81min in 10-80CD_8min.met,chromatography(XBridge Shield RP18 5um).

1H NMR(400MHz,DMSO-d₆):δ＝11.31(s,1H),9.46(s,1H),8.81(s,1H),8.07(d,J＝8.8Hz,1H),7.86-7.60(m,4H),7.35(d,J＝8.8Hz,1H),5.16-4.76(m,1H),3.03(s,3H),1.82-1.63(m,1H),1.32-1.15(m,2H).

The example compounds of the following table 5 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 5 ]

Example 13: preparation of Compound EXP-34

The synthetic route is as follows:

step 13.1 preparation of Compound 21

5-bromobenzothiophene (SM9) (200mg,0.94mmol) and morpholine (SM23) (163mg,1.88mmol) were dissolved in N, N-dimethylacetamide (4mL), to which was added cuprous iodide (18mg,0.094mmol) and sodium tert-butoxide (180mg,1.88mmol) and reacted with microwaves at 190 ℃ for 2 hours. The reaction mixture was combined with EB5-63(100mg of 5-bromobenzothiophene) and diluted with ethyl acetate (20mL) and water (20 mL). The aqueous phase was extracted with ethyl acetate (20 mL. times.3), the organic phase was washed with saturated brine (20 mL. times.3), dried over anhydrous sodium sulfate, filtered, and concentrated, followed by silica gel column chromatography (eluent: 0% -20% ethyl acetate/petroleum ether) to give 4- (benzothien-5-yl) morpholine (21) (80mg, yield 24%, purity 92%) as a pale yellow target product.

LCMS:t_R＝0.975min in 5-95AB_1.5min_220&254_Shimadzu.lcm(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝220.0[M+H]⁺.

Step 13.2 preparation of Compound 22

4- (Benzothien-5-yl) morpholine (21) (80mg,0.36mmol) was dissolved in tetrahydrofuran (2mL) and n-butyllithium (2.5M in hexane,0.44mL) was slowly added dropwise thereto at-75 ℃ under a nitrogen atmosphere. After stirring for 60 minutes, triisopropyl borate (137mg,0.73mmol) was added, then slowly raised to 15-20 ℃ and stirring continued for 16 hours. The reaction was quenched with 10% hydrochloric acid solution (10mL) and the aqueous phase extracted with ethyl acetate (10 mL. times.3), the organic phase dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give (5-morpholinobenzothien-2-yl) boronic acid (22) as a yellow solid (60mg, crude).

Step 13.3 preparation of Compound EXP-34

N- (5-bromo- [1,2, 4)]Triazole [1,5-a ]]Pyridin-2-yl]Cyclopropanecarboxamide (Int-1) (30mg,0.11mmol) and (5-morpholinobenzothien-2-yl) boronic acid (22) (42mg,0.16mmol) were dissolved in dioxane (0.5mL) and water (0.1mL), and potassium carbonate (44mg,0.32mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (8mg,0.01mmol) were added rapidly under a nitrogen atmosphere. The mixture was replaced with nitrogen three times, and then stirred at 90 ℃ for 2 hours. The reaction mixture was filtered, the filtrate was diluted with ethyl acetate (20mL) and water (20mL), the aqueous phase was extracted with ethyl acetate (20 mL. times.3), the organic phase was washed with saturated brine (20 mL. times.3), dried over anhydrous sodium sulfate, filtered, concentrated and purified by preparative HPLC (separation column: AgelaDuraShell C18150 mm. times.25 mm. times.5 um; mobile phase: A: water (0.04% NH)₃.H₂O+10mM NH₄HCO₃) Andb is acetonitrile; gradient: b40-70%) to obtain white solid N- [5- [ 5-morphine benzothiophen-2-yl)]-[1,2,4]Triazole [1,5-a ]]Pyridin-2-yl]Cyclopropanecarboxamide (EXP-34) (1.9mg, yield 4%, purity 91.88%).

LCMS:t_R＝2.719min in 10-80AB_4min_220&254_Shimadzu.lcm(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝420.3[M+H]⁺.

HPLC:t_R＝3.07min(10-80AB_8min.met),chromatography.

1H NMR(400MHz,DMSO-d₆):δ＝11.20(s,1H),8.71(s,1H),7.91(d,J＝8.8Hz,1H),7.78-7.69(m,3H),7.37(d,J＝1.6Hz,1H),7.28(dd,J＝8.8and 2.0Hz,1H),3.79(t,J＝4.8Hz,4H),3.19(t,J＝4.8Hz,4H),2.18-2.02(m,1H),0.92-0.86(m,4H).

The example compounds of the following table 6 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 6 ]

Example 14: preparation of Compound EXP-37

Step 14.1 preparation of Compound 23

To a toughened glass bottle containing 5-bromobenzothiophene (SM9) (3.74g,17.20mmol) and triethylamine (5.0mL, d ═ 0.727g/mL,35.92mmol) in absolute ethanol (99mL) and tetrahydrofuran solution (33mL) was added 1, 1' -bis (diphenylphosphino) ferrocene dichloropalladium (730mg,1.00mmol) under an argon stream. After covering the pressure gauge head on the toughened glass bottle, replacing the internal pressure with argon for several times, and then filling carbon monoxide with internal pressure of 45 psi. The reaction was stirred for 50 hours at 80 ℃ under a carbon monoxide atmosphere at an internal pressure of 45 psi. The resulting deep red solution was cooled, most of the ethanol was evaporated under reduced pressure, and the remaining N, N-dimethylformamide solution was diluted with t-butyl methyl ether (300mL), washed with water (100 mL. times.3) and saturated brine (120mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by an automated preparative chromatograph (eluent: 1-3.5% ethyl acetate/petroleum ether), the combined pure fractions were collected and concentrated to dryness to give benzothiophene-5-carboxylic acid ethyl ester (23) (3.27g, 92% yield, light yellow oil).

¹H NMR(400MHz,CDCl₃):δ＝8.54(d,J＝1.2Hz,1H),8.01(dd,J＝8.4,1.6Hz,1H),7.92(d,J＝8.4Hz,1H),7.51(d,J＝5.6Hz,1H),7.43(d,J＝5.6Hz,1H),4.42(q,J＝7.2Hz,2H),1.43(t,J＝7.2Hz,3H).

Step 14.2 preparation of Compound 24

To a solution of benzothiophene-5-carboxylic acid ethyl ester (23) (2.83g,13.72mmol) in tetrahydrofuran (16mL) was added a solution of lithium hydroxide monohydrate (1.17g,27.9mmol) in water (14mL), and the resulting mixture was warmed to 55 ℃ and stirred for 8 hours. The resulting colorless emulsion was cooled and concentrated under reduced pressure to remove most of the volatiles, acidified with 1M hydrochloric acid (29mL) under stirring, and extracted with ethyl acetate (60 mL. times.2). The extracts were combined and washed with saturated brine (60mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give benzothiophene-5-carboxylic acid (24) (2.39g, 98% yield, off-white solid) which was used directly in the next reaction.

¹H NMR(400MHz,DMSO-d₆):δ＝12.96(brs,1H),8.51(s,1H),8.12(d,J＝8.4Hz,1H),7.90(dd,J＝8.0,1.2Hz,1H),7.88(d,J＝6.0Hz,1H),7.61(d,J＝5.6Hz,1H).

Step 14.3 preparation of Compound 25

Tert-butyllithium (7.8mL,1.3M in hexane) was added dropwise continuously to a solution of benzothiophene-5-carboxylic acid (24) (597mg,3.35mmol) in anhydrous tetrahydrofuran (15mL) at-60 ℃ under a nitrogen atmosphere for 10 minutes. After the low temperature bath was removed and the reaction was allowed to naturally warm to 0 ℃ and stirring was continued for 40 minutes, the resulting yellow suspension reaction solution was cooled again to-60 ℃ and triisopropyl borate (1.2mL, d ═ 0.818g/mL) was continuously added dropwise thereto for 10 minutes, followed by allowing the reaction solution to naturally warm to room temperature (15 ℃) and stirring was continued for 15 hours. The resulting yellow solution was quenched with 0.2M dilute hydrochloric acid (60mL) and extracted with ethyl acetate (20mL,15mL and 10 mL). The combined extracts were washed with saturated brine (25mL), dried over anhydrous sodium sulfate, filtered and concentrated to dryness to give a pale red solid. The solid was slurried with a mixed n-hexane/chloroform/methanol system (18mL, vol 30/5/1), the insoluble material was collected by filtration, washed with n-hexane (3 mL. times.2) and dried under vacuum to give 2-dihydroxybenzothiophene-5-carboxylic acid (25) (505mg, about 90% purity, 61% yield, off-white solid) which was used directly in the next reaction.

¹H NMR(400MHz,DMSO-d₆):δ＝12.98(brs,1H),8.59(brs,2H),8.52-8.46(m,1H),8.08(d,J＝8.4Hz,1H),8.06(s,1H),7.93-7.87(m,1H).

Step 14.4 preparation of Compound EXP-37

To a mixed solution of 1, 4-dioxane (10mL) and water (2mL) of N- (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -cyclopropanecarboxamide (Int-1) (369mg,1.31mmol) and 2-dihydroxyborobenzothiophene-5-carboxylic acid (25) (505mg, about 90% purity, 2.05mmol) was added potassium phosphate (822mg,3.87mmol) and 1, 1' -bis (diphenylphosphino) ferrocene palladium dichloride (32mg,0.043umol) under a nitrogen stream. The mixture was replaced with nitrogen several times, and then heated to 95 ℃ under nitrogen atmosphere and stirred continuously for 20 hours. The resulting dark brown suspension (containing pilot batch EB3-34) was cooled, acidified to pH 3 with 1M hydrochloric acid and extracted with ethyl acetate (25mL,20mL and 15 mL). The combined extracts were washed with saturated brine (30mL), dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The crude product is first purified by an automatic preparative chromatograph (eluent: 80-99% ethyl acetate/petroleum ether, then 1-8% methanol/dichloromethane). The desired fractions were collected and evaporated to dryness under reduced pressure to give the crude product as a pale yellow solid (155 mg).

LCMS:t_R＝0.946min in 5-95 AB_1.6min_220&254_Shimadzu.lcm,MS(ESI)m/z＝401.0[M+Na]⁺.

t_R＝3.973min in 10-80 AB_7min_220&254_Shimadzu.lcm,MS(ESI)m/z＝379.2[M+H]⁺.

73 mg of the above solid was isolated by HPLC (25-55% acetonitrile/0.225% formic acid), the pure fractions were collected, concentrated under reduced pressure and lyophilized to give the final product 2- [2- (cyclopropylcarboxamido) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl ] benzothiophene-5-carboxylic acid (EXP-37) (6.5mg, 93% purity, 2% yield, off-white solid).

LCMS:t_R＝3.787min in 10-80 AB_7min_220&254_Shimadzu.lcm,MS(ESI)m/z＝379.2[M+H]⁺.

HPLC:t_R＝3.79min in 0-60 AB_8min.met.

¹H NMR(400MHz,DMSO-d₆):δ＝11.22(brs,1H),8.89(s,1H),8.55(s,1H),8.21(d,J＝8.4Hz,1H),8.00(d,J＝8.4Hz,1H),7.88-7.70(m,3H),2.18-1.93(m,1H),0.91-0.84(m,4H).

Example 15: preparation of Compound EXP-38

The synthetic route is as follows:

step 15.1 preparation of Compound 26

To a toughened glass bottle containing 6-bromobenzothiophene (SM6) (935mg,4.30mmol) and triethylamine (1.3mL, d ═ 0.727g/mL,9.34mmol) in anhydrous ethanol (25mL) and tetrahydrofuran solution (10mL) was added 1, 1' -bis (diphenylphosphino) ferrocene dichloropalladium (170mg,0.232mmol) under an argon stream. After covering the pressure gauge head on the toughened glass bottle, the internal pressure was replaced several times with argon gas, and then carbon monoxide with an internal pressure of 50psi was introduced. The reaction was stirred for 40 hours at 80 ℃ under a carbon monoxide atmosphere at an internal pressure of 50 psi. The resulting deep red solution was cooled, most of the ethanol was evaporated under reduced pressure, and the remaining N, N-dimethylformamide solution was diluted with t-butyl methyl ether (90mL), washed with water (30 mL. times.3) and saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by an automated preparative chromatograph (eluent: 1-3% ethyl acetate/petroleum ether), the combined pure fractions were collected and concentrated to dryness to give benzothiophene-6-carboxylic acid methyl ester (26) (821mg, 99% yield, white solid).

¹H NMR(400MHz,CDCl₃):δ＝8.62(s,1H),8.03(d,J＝8.0Hz,1H),7.86(d,J＝8.4Hz,1H),7.65(d,J＝5.6Hz,1H),7.40(d,J＝5.6Hz,1H),3.96(s,3H).

Step 15.2 preparation of Compound 27

Methyl benzothiophene-6-carboxylate (26) (821mg,4.27mmol), pinacolborane (SM10) (0.75mL, d ═ 0.882g/mL,4.91mmol), methoxy (cyclooctadiene) iridium (I) dimer (45mg,0.064mmol) and a mixture of 4,4 '-di-tert-butyl-2, 2' -bipyridine (39mg,0.141mmol) in n-hexane (24mL) were replaced with a nitrogen atmosphere several times and stirring was continued at 15 ℃ for 12 hours. The resulting red solution was filtered to remove insoluble matter and concentrated under reduced pressure. The residue was purified by an automatic preparative chromatograph (eluent: 1-4% ethyl acetate/petroleum ether), the combined pure fractions were collected and concentrated to dryness to give 6-methoxycarbonylbenzothiophene-2-boronic acid pinacol ester (27) (450mg, 33% yield, off-white solid).

¹H NMR(400MHz,DMSO-d₆):δ＝8.63(s,1H),8.00(dd,J＝8.4,1.6Hz,1H),7.91(s,1H),7.88(d,J＝8.4Hz,1H),3.96(s,3H),1.38(s,12H).

Step 15.3 preparation of Compound 28

To a 28mL microwave tube were added N- (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -cyclopropanecarboxamide (Int-1) (183mg,0.651mmol) and 6-methoxycarbonylbenzothiophene-2-boronic acid pinacol ester (27) (290mg,0.911mmol) in that order in a mixed solution of 1, 4-dioxane (5.5mL) and water (1mL), sodium carbonate (138mg,1.30mmol) was added, followed by bis (triphenylphosphine) palladium dichloride (59mg,0.085mmol) under a nitrogen stream. After the nitrogen atmosphere was replaced by bubbling in the microwave tube, the tube was sealed and placed in a microwave reactor, and the reaction was continued for 2.5 hours at 110 ℃. The resulting orange suspension was cooled and concentrated under reduced pressure to remove most of the volatiles. The residue was dissolved in dichloromethane/methanol (50mL, volume 4/1) and the insolubles were removed by filtration. The filtrate was concentrated under reduced pressure to dryness, and the resulting crude product (containing a pilot batch EB3-71) was purified by an autopreparation chromatograph (eluent: 0-25% acetone/(petroleum ether/ethyl acetate 1/1 vol%). The desired fractions were collected and evaporated to dryness under reduced pressure to give methyl 2- [2- (cyclopropylcarboxamido) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl ] benzothiophene-6-carboxylate (28) (90mg, 60% purity, 19% yield, pink solid).

LCMS:t_R＝1.073min in 5-95 AB_1.6min_220&254_Shimadzu.lcm,MS(ESI)m/z＝393.1[M+H]⁺.

t_R＝3.580min in 10-80 AB_7min_220&254_Shimadzu.lcm,MS(ESI)m/z＝393.2[M+H]⁺.

Step 15.4 preparation of Compound 29

To a solution of methyl 2- [2- (cyclopropylcarboxamido) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl ] benzothiophene-6-carboxylate (28) (90mg, about 60% pure, 0.138mmol) in tetrahydrofuran (1.4mL) was added a solution of lithium hydroxide monohydrate (29mg,0.691mmol) in water (0.6 mL). The reaction solution was stirred continuously at 30 ℃ for 12 hours. The resulting brown solution was concentrated to remove most of the volatiles and acidified to pH 3 with 1M hydrochloric acid solution. The resulting precipitate was collected by filtration and washed with water (0.5 mL. times.2), then dried in vacuo to give 2- [2- (cyclopropylcarboxamido) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl ] benzothiophene-6-carboxylic acid (29) (66mg, about 50% purity, 63% yield, pink solid) and used directly in the next reaction.

LCMS:t_R＝1.028min in 5-95 AB_2min_220&254_Shimadzu.lcm,MS(ESI)m/z＝379.2[M+H]⁺.

Step 15.5 preparation of Compound EXP-38

To a solution of 2- [2- (cyclopropylcarboxamido) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl ] benzothiophene-6-carboxylic acid (29) (66mg, about 50% purity, 0.087mmol) in anhydrous N, N-dimethylformamide (1.4mL) were added O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium hexafluorophosphate (50mg,0.132mmol) and N, N-diisopropylethylamine (0.05mL, d ═ 0.742g/mL,0.287mmol) in that order, and (2-thiazol-2-yl) ethylamine (17mg,0.133mmol) was added later. After the reaction mixture was stirred at 15 ℃ for 6 hours, the resulting yellow slurry was diluted with ethyl acetate (15mL), washed with water (5 mL. times.2) and saturated brine (5mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was chromatographed using high performance liquid chromatography (25-55% acetonitrile/0.04% ammonia) to give pure fractions, which were concentrated under reduced pressure and lyophilized to give the final product 2- [2- (cyclopropylcarboxamido) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl ] -N- (2-thiazol-2-yl) -ethyl ] benzothiophene-6-carboxamide (EXP-38) (4.8mg, 100% purity, 11% yield, white solid).

LCMS:t_R＝2.820min in 10-80 AB_7min_220&254_Shimadzu.lcm,MS(ESI)m/z＝489.3[M+H]⁺.

t_R＝2.860min in 10-80 AB_7min_220&254_Shimadzu.lcm,MS(ESI)m/z＝489.3[M+H]⁺.

HPLC:t_R＝2.55min in 10-80 AB_8min.met.

¹H NMR(400MHz,Methanol-d₄):δ＝8.86(s,1H),8.40(s,1H),8.01(d,J＝8.4Hz,1H),7.84(d,J＝8.4,1.2Hz,1H),7.78-7.69(m,3H),7.67(dd,J＝7.6and 2.0Hz,1H),7.49(d,J＝3.2Hz,1H),3.82(t,J＝6.8Hz,2H),3.41(t,J＝6.8Hz,2H),2.13-1.92(m,1H),1.15-1.05(m,2H),1.03-0.93(m,2H).

The example compounds of the following table 7 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 7 ]

Example 16: preparation of Compound EXP-41

The synthetic route is as follows:

step 16.1 preparation of Compound 30

[5- (Methylsulfonamido) benzothiophen-2-yl ] boronic acid (3) (1.1g, 4.1mmol), 5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-amine (SM12) (432mg, 2.0mmol) and sodium carbonate (430mg, 4.1mmol) were dissolved in water (5mL) and tetrahydrofuran (20mL) and nitrogen-substituted for 3 times, then 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (285mg, 0.41mmol) was added and nitrogen-substituted for 3 times, and the reaction was carried out at 100 ℃ for 1 hour. Ethyl acetate (10mL) and water (10mL) were added to dilute the solution, followed by extraction with ethyl acetate (10 mL. times.3), drying over anhydrous sodium sulfate, filtration, concentration in vacuo, and purification by column chromatography to give N- (2- (2-amino- [1,2-4] triazolo [1,5-a ] pyridin-5-yl) benzothien-5-yl ] methanesulfonamide (30) (700 mg, crude) as a yellow solid.

Step 16.2 preparation of Compound 31

N- [2- (2-amino- [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzothien-5-yl ] methanesulfonamide (30) (200mg,0.56 mmol) and sodium nitrite (77mg, 1.1mmol) were dissolved in acetonitrile (2 mL). After stirring at 0 ℃ for 0.1 hour under nitrogen, hydrogen bromide (287mg, 1.7mmol) was added dropwise and the mixture was stirred at 20 ℃ for 0.9 hour. Water (5mL) was added, saturated potassium carbonate was adjusted to pH 7, and dichloromethane (10mL × 3) was extracted. Washed with saturated brine (10mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give N- [2- (2-bromo- [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzothien-5-yl ] methanesulfonamide (31) (150mg, crude) as a yellow solid.

Step 16.3 preparation of Compound EXP-41

N- [2- (2-bromo- [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzothien-5-yl ] methanesulfonamide (31) (20mg, 0.05mmol), 1-methylpyrazol-3-amine (9mg, 0.09mmol), Brettphos Pd G3(9mg, 0.01mmol) and sodium tert-butoxide (36mg, 0.38mmol) were dissolved in 1, 4-dioxane (2 mL). Heating for 0.2 hour at 150 deg.C with microwave. Ethyl acetate (10mL) and water (10 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was isolated as a yellow solid N- [2- [2- [ (1-methylpyrazol-3-yl) amino ] - [1,2,4] triazolo [1,5-a ] pyridin-5-yl ] benzothien-5-yl ] methanesulfonamide (EXP-41) (6.7mg, 30% yield).

LCMS:t_R＝2.133min in 10-80AB_4min_220&254_Agilent.met,chromatography(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝440.3[M+H]⁺.

HPLC:t_R＝2.16min in 10-80CD_8min.met,chromatography(XBridge C18 2.1*50mm 5um).

1H NMR(400MHz,DMSO-d₆):δ＝11.25(s,1H),9.87(s,1H),8.81(s,1H),8.05(d,J＝8.8Hz,1H),7.86-7.65(m,4H),7.34(d,J＝8.8Hz,1H),6.83(s,1H),3.84(s,3H),3.10(s,3H).

The example compounds of the following table 8 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 8 ]

Example 17: preparation of Compound EXP-44

The synthetic route is as follows:

step 17.1 preparation of Compound 32

5-bromobenzothiophene (SM9) (4g, 18.77mmol), potassium vinyltrifluoroborate salt (5.03g,37.54mmol), 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (686mg, 0.938mmol) and cesium carbonate (15.29g, 46.93mmol) were dissolved in water (4 mL)/dioxane (40mL) and stirred at 90 degrees for 2 hours after nitrogen exchange three times. The mixture was added water (50mL) and extracted with dichloromethane (50 mL. times.3). The organic phase was washed once with saturated brine (80mL), then dried over anhydrous sodium sulfate and concentrated by filtration. The crude product obtained was subjected to silica gel column chromatography (eluent: 1% ethyl acetate in petroleum ether) to give 5-vinylbenzothiophene (32) (2.97g, yield 98%) as a colorless oil.

1H NMR(400MHz,DMSO-d₆):δ＝7.87-7.80(m,2H),7.49(dd,J＝8.4,2.0Hz,1H),7.45(d,J＝5.6Hz,1H),7.34(d,J＝5.2Hz,1H),6.92-6.80(m,1H),5.83(d,J＝17.2Hz,1H),5.30(d,J＝11.2Hz,1H).

Step 17.2 preparation of Compound 33

5-vinyl benzothiophene (32) (2.97g, 18.54mmol) was dissolved in tetrahydrofuran (60mL) and 9-borabicyclo [3.3.1] nonane (0.5M in tetrahydrofuran, 55.6mL) was added at 0 deg.C, the reaction was stirred at 10-15 deg.C for 16 hours and then water (6mL), 3M sodium hydroxide solution (30mL) and hydrogen peroxide (30mL, 30 wt%) were added, and the reaction was stirred at 50 deg.C for 2 hours. The mixture was quenched with saturated sodium sulfite (40mL) and extracted with ethyl acetate (60 mL. times.3), and the organic phase was washed once with saturated brine (80mL), then dried over anhydrous sodium sulfate and concentrated by filtration. The crude product obtained was chromatographed on silica gel (eluent: 50% ethyl acetate in petroleum ether) to give 2- (benzothien-5-yl) ethanol (33) (2.4g, yield 72%) as a white solid.

1H NMR(400MHz,DMSO-d₆):δ＝7.84(d,J＝8.4Hz,1H),7.70(d,J＝0.8Hz,1H),7.45(d,J＝5.6Hz,1H),7.31(d,J＝5.2Hz,1H),7.24(dd,J＝8.4,1.6Hz,1H),3.93(t,J＝6.0Hz,2H),3.00(t,J＝6.4Hz,2H),1.48-1.40(m,1H).

Step 17.3 preparation of Compound 34

2- (Benzothien-5-yl) ethanol (33) (1g, 5.61mmol) was dissolved in tetrahydrofuran (10mL) and after addition of sodium hydrogen (448mg, 11.22mmol) at 0 degrees and stirring at 15 degrees for half an hour, 4-methylbenzenesulfonyl chloride (1.28g, 6.73mmol) was added to the reaction and stirred at 10-15 degrees for 1.5 hours. The mixture was quenched with water (25mL), extracted with ethyl acetate (40 mL. times.2), and the organic phase was washed once with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product obtained was subjected to silica gel column chromatography (eluent: 5% ethyl acetate/petroleum ether) to give 2- (benzothien-5-yl) ethyl 4-methylbenzenesulfonic acid (34) as a white solid (1.4g, yield 75%).

1H NMR(400MHz,DMSO-d₆):δ＝7.74(d,J＝8.0Hz,1H),7.61(d,J＝8.4Hz,2H),7.54(d,J＝0.8Hz,1H),7.45(d,J＝5.6Hz,1H),7.24(d,J＝5.6Hz,1H),7.16(d,J＝8.0Hz,2H),7.08(dd,J＝8.4,1.6Hz,1H),4.27(t,J＝6.8Hz,2H),3.07(t,J＝6.8Hz,2H),2.39(s,3H).

Step 17.4 preparation of Compound 35

2- (Benzothien-5-yl) ethyl 4-methylbenzenesulfonic acid (34) (400mg,1.20mmol) was dissolved in N, N-dimethylformamide (5mL) and potassium carbonate (498mg,3.61mmol) and thiomorpholine-1, 1-dioxide (406mg,3.01mmol) were added and stirred at 85 ℃ for 2 hours. The mixture was added with ethyl acetate (20mL), washed 3 times with saturated brine (20mL), dried over anhydrous sodium sulfate, and concentrated by filtration. The crude product obtained was subjected to silica gel column chromatography (eluent: 50% ethyl acetate/petroleum ether) to give 4- [2- (benzothien-5-yl) ethyl ] -thiomorpholine-1, 1-dioxide (35) as a white solid (80mg, yield 19%).

LCMS:t_R＝0.898min in 10-80AB_2.0min_220&254_Shimadzu.lcm chromatography(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝296.1[M+H]⁺.

Step 17.5 preparation of Compound 36

4- [2- (benzothien-5-yl) ethyl ] -thiomorpholine-1, 1-dioxide (35) (70mg,0.236mmol) was dissolved in tetrahydrofuran (2mL) and after cooling to-75 ℃ under a nitrogen atmosphere, n-butyllithium (2.5M in n-hexane, 0.28mL) was added slowly and stirred at-75 ℃ for half an hour, triisopropyl borate (133mg,0.71mmol) was added and the mixture was raised to 15-20 ℃ and stirred for 2 hours. The mixture was quenched with 1M hydrochloric acid (10mL) and extracted with ethyl acetate (20 mL. times.3), and the organic phase was washed once with saturated brine (20mL), then dried over anhydrous sodium sulfate and concentrated by filtration to give [5- [2- (1, 1-dioxide-1, 4-thiomorpholin-4-yl) ethyl ] benzothien-2-yl ] phenylboronic acid (36) (70mg, 87% yield).

LCMS:t_R＝0.788min in 10-80AB_2.0min_220&254_Shimadzu.lcm chromatography(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝340.1[M+H]⁺.

Step 17.6 preparation of Compound EXP-44

N- (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (Int-1) (58mg,0.206mmol), [5- [2- (1, 1-dioxide-1, 4-thiomorpholin-4-yl) ethyl ] benzothien-2-yl ] phenylboronic acid (36) (70mg,0.206mmol), potassium phosphate (109mg,0.515mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (7mg,0.01mmol) were dissolved in dioxane (2 mL)/water (0.2mL) and stirred at 90 ℃ for 2 hours after replacement with nitrogen three times. The reaction mixture was added with water (30mL) and extracted with dichloromethane (30 mL. times.3), and the organic phase was washed once with saturated brine (50mL) and then dried over anhydrous sodium sulfate and concentrated by filtration. The resulting crude product was subjected to silica gel column chromatography (eluent: 5% methanol/dichloromethane) to give a crude product, which was then isolated and purified by basic preparative separation to give the desired product N- [5- [2- (1, 1-dioxide-1, 4-thiomorpholin-4-yl) ethyl ] benzothien-2-yl ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (EXP-44) as a white solid (9.2mg, 9% yield).

LCMS:t_R＝2.360min in 10-80AB_7min_220&254_Shimadzu.lcm chromatography(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝496.4[M+H]⁺.

HPLC:t_R＝2.76min in 10-80CD_8min.met,chromatography(XBridge Shield RP18 5um).

1H NMR(400MHz,DMSO-d₆):δ＝11.21(s,1H),8.80(s,1H),7.99(d,J＝8.0Hz,1H),7.84(s,1H),7.80-7.67(m,3H),7.39(d,J＝7.6Hz,1H),3.13-3.06(m,4H),3.05-2.96(m,4H),2.95-2.85(m,2H),2.84-2.75(m,2H),2.18-2.03(m,1H),0.96-0.82(m,4H).

Example 18: preparation of Compound EXP-41

Step 18.1 preparation of Compound 37

2- (Benzothien-5-yl) ethyl 4-methylbenzenesulfonic acid (34) (320mg,0.962mmol) was dissolved in acetonitrile (5mL) and potassium carbonate (399mg,2.89mmol) and 4, 4-difluoropiperidine (SM8) (233mg,1.93mmol) were added and stirred at 85 ℃ for 16 hours. The mixture was added with water (20mL), extracted with ethyl acetate (20 mL. times.3), and the organic phase was washed once with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product obtained was subjected to silica gel column chromatography (eluent: 10% ethyl acetate/petroleum ether) to give 1- [2- (benzothien-5-yl) ethyl ] -4, 4-difluoropiperidine (37) as a white solid (300mg, yield 82%).

LCMS:t_R＝0.985min in 5-95AB_1.5min_220&254_Shimadzu.lcm chromatography(Xtimate C18,3um,2.1*30mm),MS(ESI)m/z＝282.0[M+H]⁺.

Step 18.2 preparation of Compound 38

1- [2- (benzothien-5-yl) ethyl ] -4, 4-difluoropiperidine (37) (65mg,0.231mmol) was dissolved in tetrahydrofuran (2mL) and after cooling to-75 ℃ under a nitrogen atmosphere, n-butyllithium (2.5M in n-hexane, 0.277mL) was added slowly and stirred for half an hour at-75 ℃ and after addition of triisopropyl borate (130mg,0.693mmol) was raised to 15-20 ℃ and stirred for 2 hours. The mixture was quenched with 1M hydrochloric acid (10mL) and extracted with ethyl acetate (20 mL. times.3), and the organic phase was washed once with saturated brine (20mL), then dried over anhydrous sodium sulfate and concentrated by filtration to give [5- [2- (4, 4-difluoropiperidine) ethyl ] benzothien-2-yl ] phenylboronic acid (38) (70mg, yield 93%) as a white solid.

LCMS:t_R＝0.197min in 5-95AB_4.0min_220&254_Shimadzu.lcm chromatography(Xtimate,2.1*30mm,3um B:XBrige Shield 2.1*50mm),MS(ESI)m/z＝326.1[M+H]⁺.

Step 18.3 preparation of Compound EXP-45

N- (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (38) (60mg,0.215mmol), [5- [2- (4, 4-difluoropiperidine) ethyl ] benzothien-2-yl ] phenylboronic acid (70mg,0.215mmol), potassium phosphate (114mg,0.538mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (7mg,0.01mmol) were dissolved in dioxane (2 mL)/water (0.2mL) and stirred at 90 ℃ for 2 hours after replacing nitrogen three times. The reaction mixture was added with water (30mL) and extracted with dichloromethane (30 mL. times.3), and the organic phase was washed once with saturated brine (50mL) and then dried over anhydrous sodium sulfate and concentrated by filtration. The obtained crude product was subjected to silica gel column chromatography (eluent: 5% methanol/dichloromethane) to obtain a crude product, which was then isolated and purified by basic preparation to obtain the aimed product N- [5- [5- [2- (4, 4-difluoropiperidine) ethyl ] benzothien-2-yl ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (EXP-45) as a white solid (8.5mg, yield 8%).

LCMS:t_R＝2.494min in 10-80AB_7min_220&254_Shimadzu.lcm chromatography(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝482.4[M+H]⁺.

HPLC:t_R＝3.41min in 10-80CD_8min.met,chromatography(XBridge Shield RP 18 5um).

1H NMR(400MHz,DMSO-d₆):δ＝11.21(s,1H),8.79(s,1H),7.98(d,J＝8.4Hz,1H),7.83(s,1H),7.79-7.68(m,3H),7.37(dd,J＝8.0,0.8Hz,1H),2.96-2.85(m,2H),2.70-2.65(m,2H),2.63-2.58(m,4H),2.16-2.05(m,1H),2.0-1.85(m,4H),0.94-0.84(m,4H).

The example compounds of the following table 9 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 9 ]

Example 19: preparation of Compound EXP-48

The synthetic route is as follows:

step 19.1 preparation of Compound 39

5-bromobenzothiophene (SM9) (1.5g,7.04mmol) was dissolved in dimethylformamide (13.5mL) and pyridine (0.75mL), and cuprous cyanide (851mg,9.50mmol) was added. The reaction solution was stirred at 150 ℃ for 20 hours under a nitrogen atmosphere. The reaction solution was poured into a solution of N, N-dimethylethylenediamine (1mL) in water (20 mL). The aqueous phases were combined with ethyl acetate (80 mL. times.3), the organic phases were washed with brine (20 mL. times.3), dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and the crude product was purified by silica gel column (ethyl acetate/petroleum ether, from 0% to 5%) to give 5-cyanobenzothiophene (39) (579mg, 51% yield, 99% purity, yellow solid).

1H NMR(400MHz,DMSO-d₆):δ＝8.43(d,J＝0.8Hz,1H),8.25(d,J＝8.0Hz,1H),7.99(d,J＝5.6Hz,1H),7.72(dd,J＝8.4,1.6Hz,1H),7.58(d,J＝5.6Hz,1H).

Step 19.2 preparation of Compound 40

Lithium aluminum hydride (552mg,14.55mmol) was dissolved in tetrahydrofuran (20mL), a solution of 5-cyanobenzothiophene (579mg,3.64mmol) in tetrahydrofuran (10mL) was added at zero degrees, and the reaction was stirred at room temperature for 16 hours. The reaction mixture was quenched by addition of water (0.6mL) and sodium hydroxide solution (15 wt%, 0.6mL) and water (1.8mL) at zero, stirred for 0.5 h, dried over anhydrous sodium sulfate and added ethyl acetate (100 mL). The reaction was filtered, the organic phase washed with brine (20mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to afford benzothien-5-ylmethylamine (40) (590mg, 90% yield, 91% purity, yellow solid) which was used directly in the next step.

1H NMR(400MHz,DMSO-d₆):δ＝7.90(d,J＝8.4Hz,1H),7.81(d,J＝0.4Hz,1H),7.72(d,J＝5.2Hz,1H),7.41(d,J＝5.2Hz,1H),7.34(dd,J＝8.4,1.2Hz,1H),3.82(s,2H).

Step 19.3 preparation of Compound 41

Benzothien-5-ylmethylamine (40) (590mg,3.61mmol) was dissolved in dichloromethane (10mL), triethylamine (1.28g,12.64mmol) and methanesulfonic anhydride (1.26g,7.22mmol) were added at zero degrees, and the reaction was stirred at room temperature for 16 hours. The reaction solution was quenched by addition of saturated sodium bicarbonate (20mL) at zero degrees and extracted with dichloromethane (50 mL. times.3). The organic phases were combined, washed with brine (50mL), dried over anhydrous sodium sulfate, filtered, concentrated in vacuo and the crude product was purified by silica gel chromatography (ethyl acetate/petroleum ether, from 0% to 39%) to give N- (benzothien-5-ylmethyl) methanesulfonamide (41) (655mg, 75% yield, as a pale yellow solid).

1H NMR(400MHz,DMSO-d₆):δ＝7.98(d,J＝8.4Hz,1H),7.84(s,1H),7.77(d,J＝5.2Hz,1H),7.61(t,J＝6.4Hz,1H),7.46(d,J＝5.6Hz,1H),7.35(dd,J＝8.4,1.6Hz,1H),4.27(d,J＝6.4Hz,2H),2.86(s,3H).

Step 19.4 preparation of Compound 42

N- (benzothien-5-ylmethyl) methanesulfonamide (41) (200mg,0.829mmol) was dissolved in tetrahydrofuran (4mL) and N-butyllithium (2.5M in hexane,1.33mL) was added under nitrogen protection at-78 ℃ for stirring for 0.5 hour, and triisopropyl borate (655mg,3.48mmol) was added thereto, and the reaction mixture was stirred at-78 ℃ for 0.5 hour, warmed to room temperature and stirred for 2 hours. The reaction was quenched with dilute hydrochloric acid (1M,5mL) at zero degrees and stirred at room temperature for half an hour. The reaction solution was extracted with ethyl acetate (40 mL. times.3). The organic phases were combined, washed with brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give [5- (methanesulfonamidomethyl) benzothien-2-yl ] boronic acid (42) (260mg, crude, yellow solid) which was used directly in the next step.

Step 19.5 preparation of Compound EXP-48

N- (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropylcarboxamide (Int-) (100mg,0.356mmol), potassium carbonate (98mg,0.711mmol) and [5- (methanesulfonamido methyl) benzothien-2-yl ] boronic acid (42) (142mg,0.498mmol) were dissolved in dioxane (2mL) and water (0.4mL), displaced with nitrogen three times, then 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (39mg,0.0534mmol) was added, displaced with nitrogen three times, and the reaction was stirred at 95 ℃ for 2 hours. The reaction was concentrated to give a crude product which was purified by silica gel chromatography (methanol/dichloromethane, from 0% to 3%) to give N- [5- [5- (methanesulfonamido methyl) benzothien-2-yl ] - [1,2,4] triazole [1,5-a ] pyridin-2-yl ] cyclopropylcarboxamide (EXP-48) (30.2mg, 19% yield, 98.30% purity, yellow solid).

LCMS:t_R＝2.890min in 10-80AB_7min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*3mm,3um),MS(ESI)m/z＝442.3[M+H]⁺.

HPLC:t_R＝3.09min in 10-80AB_8min_215&220&254.met.

1H NMR(400MHz,DMSO-d₆) δ 11.23(brs,1H),8.86(s,1H),8.07(d, J8.4 Hz,1H),7.94(s,1H),7.79-7.71(m,3H),7.68(t, J6.4 Hz,1H),7.47(dd, J8.4, 1.2Hz,1H),4.32(d, J6.4 Hz,2H),2.90(s,3H),2.20-2.02(m,1H),0.95-0.82(m,4H). example 20: preparation of Compound EXP-49

The synthetic route is as follows:

step 20.1 preparation of Compound 43

Benzothiophene-5-amine (1) (200mg,1.34mmol), triethylamine (406mg,4.02mmol) and 4-dimethylaminopyridine (16mg,0.13mmol) were dissolved in dichloromethane (5mL), cyclopropylsulfonyl chloride (SM16) (282mg,2.01mmol) was added at 0 ℃ and stirred at 15-25 ℃ for 2 hours. The reaction solution was concentrated and spin-dried by silica gel column chromatography (eluent: 0% to 50% ethyl acetate/petroleum ether) to give N- (benzothien-5-yl) cyclopropanesulfonamide (43) (200mg, yield 59%) as a white target product.

1H NMR(400MHz,DMSO-d₆):δ＝9.74(s,1H),7.94(d,J＝8.4Hz,1H),7.81-7.69(m,2H),7.44(d,J＝5.6Hz,1H),7.31-7.23(m,1H),2.65-2.55(m,1H),0.97-0.82(m,4H).

Step 20.2 preparation of Compound 44

N- (benzothien-5-yl) cyclopropanesulfonamide (43) (200mg,0.79mmol) was dissolved in tetrahydrofuran (3mL), to which N-butyllithium (2.5M in hexane,1.26mL) was slowly added dropwise at-75 ℃ under a nitrogen atmosphere. After stirring for 60 min, triisopropyl borate (297mg,1.58mmol) was added, then slowly raised to 15-20 ℃ and stirring continued for 2 h. The reaction was quenched with 10% hydrochloric acid solution (20mL) and the aqueous phase was extracted with ethyl acetate (20mL × 3), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to give [5- (cyclopropanesulfonamide) benzothien-2-yl ] boronic acid (44) (200mg, crude) as a pale yellow solid.

Step 20.3 preparation of Compound EXP-49

N- (5-bromo- [1,2, 4)]Triazole [1,5-a ]]Pyridin-2-yl]Cyclopropanecarboxamide (Int-1) (80mg,0.28mmol) and [5- (Cyclopropanesulfonamide) benzothien-2-yl]Boric acid (44) (169mg,0.57mmol) was dissolved in dioxane (3mL) and water (0.6mL), and potassium carbonate (118mg,0.85mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (20mg,0.028mmol) were added rapidly under a nitrogen atmosphere. The mixture was replaced with nitrogen three times, and then stirred at 90 ℃ for 2 hours. The reaction mixture was filtered, the filtrate was diluted with dichloromethane (20mL) and water (20mL), the aqueous phase was extracted with dichloromethane (20 mL. times.3), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and dried, and purified by silica gel column chromatography (eluent: 0% to 10% methanol/dichloromethane) and HPLC (separation column: Waters Xbridge 150 mm. times.25 mm. times.5 um; mobile phase: A: water (0.04% NH. times.5 um)₃.H₂O+10mM NH₄HCO₃) And B is acetonitrile; gradient: b30-60%) to yield N- [5- [5- (cyclopropylsulfonamide) benzothien-2-yl ] white solid]-[1,2,4]Triazole [1,5-a ]]Pyridin-2-yl]Cyclopropanecarboxamide (EXP-49) (10.4mg, yield 7%, purity 84.89%).

LCMS:t_R＝2.188min in 10-80AB_4min_220&254_Shimadzu.lcm(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝454.30[M+H]⁺.

HPLC:t_R＝3.35min in 10-80AB_8min.met,chromatography.

1H NMR(400MHz,DMSO-d₆):δ＝11.22(s,1H),9.82(s,1H),8.81(s,1H),8.05(d,J＝8.8Hz,1H),7.83(d,J＝2.0Hz,1H),7.79-7.70(m,3H),7.38(dd,J＝8.8,2.0Hz,1H),2.74-2.61(m,1H),2.18-2.01(m,1H),0.97-0.83(m,8H).

Example 21: preparation of Compound EXP-50

The synthetic route is as follows:

step 21.1 preparation of Compound 45

2- [5- (bromomethyl) benzothiophene-pinacol borate (14) (200mg,0.566mmol) was dissolved in acetonitrile (3mL), potassium carbonate (156mg,1.13mmol) and 1- (2-hydroxyethyl) piperazine (1A) (147mg,1.13mmol) were added and stirred at 35 for 2 hours. The mixture was added with water (20mL), extracted with ethyl acetate (30 mL. times.3), and the organic phase was washed once with saturated brine (30mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 2- [4- [ [2- (pinacolboronic acid ester-2-yl) benzothiophen-5-yl ] methylpiperazin-1-yl ] ethanol (45) as a red solid (80mg, yield 35%).

LCMS:t_R＝0.837min in 5-95AB_2.0min_220&254_Shimadzu.lcm chromatography(Xtimate C18,3um,2.1*30mm),MS(ESI)m/z＝403.2[M+H]⁺.

Step 21.2 preparation of Compound EXP-50

N- (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (45) (50mg,0.177mmol),2- [4- [ [2- (pinacolboronic acid ester-2-yl) benzothien-5-yl ] methylpiperazin-1-yl ] ethanol (71mg,0.177mmol), potassium phosphate (75mg,0.355mmol) and 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (6mg,0.008mmol) were dissolved in dioxane (1 mL)/water (0.1mL) and stirred at 70 ℃ for 2 hours after exchanging nitrogen three times. The reaction mixture was added with water (30mL) and extracted with dichloromethane (30 mL. times.3), and the organic phase was washed once with saturated brine (50mL) and then dried over anhydrous sodium sulfate and concentrated by filtration. The obtained crude product was subjected to silica gel column chromatography (eluent: 10% methanol/dichloromethane) to obtain a crude product, which was then isolated and purified by basic preparation to obtain N- [5- [5- [ [4- (2-hydroxyethyl) piperazin-1 yl) methyl ] benzothien-2-yl ] - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (EXP-50) (2.0mg, yield 2.0%) as a white solid.

LCMS:t_R＝1.539min in 10-80AB_4min_220&254_Shimadzu.lcm chromatography(Xtimate C18,3um,2.1*3mm),MS(ESI)m/z＝477.3[M+H]⁺.

HPLC:t_R＝2.40min in 10-80CD_8min.met,chromatography(XBridge Shield RP18 5um).

1H NMR(400MHz,DMSO-d₆):δ＝11.20(s,1H),8.81(s,1H),8.02(d,J＝8.0Hz,1H),7.86(s,1H),7.80-7.68(m,3H),7.42(dd,J＝8.4,0.8Hz,1H),4.39(s,1H),3.61(s,2H),3.49-3.47(m,2H),2.48-2.34(m,10H),2.18-2.05(m,1H),0.93-0.84(m,4H).

Example 22: preparation of Compound EXP-51

The synthetic route is as follows:

step 22.1 preparation of Compound 51

6-Bromopyridin-2-amine (SM13) (5g,28.90mmol) and ethyl bromopyruvate (SM14) (6.20g,31.79mmol) were dissolved in ethanol (30mL) and stirred at 85 ℃ for 16 h. The reaction was filtered, the filter cake collected, slurried with petroleum ether/ethyl acetate (5/1, v/v,50mL), the solid filtered and dried to give methyl 5-bromoimidazo [1,2-a ] pyridine-2-carboxylate (51) (8.41g, 83% yield, hydrobromide, yellow solid).

¹H NMR(400MHz,DMSO-d₆):δ＝8.60-8.50(m,1H),7.82-7.76(m,1H),7.65-7.47(m,2H),4.38(q,J＝7.2Hz,2H),1.35(t,J＝7.2Hz,3H).

Step 22.2 preparation of Compound 52

Methyl 5-bromoimidazo [1,2-a ] pyridine-2-carboxylate (51) (2g,5.71mmol, hydrogen bromide) is dissolved in 5ml each of water, tetrahydrofuran and methanol, and lithium hydroxide monohydrate (839mg,20.00mmol) is added. The reaction solution was stirred at 15 ℃ for 2 hours. Water (10mL) was added to the reaction mixture, and methylene chloride (20mL) was used. The aqueous phase was adjusted to pH 5 with dilute hydrochloric acid (1N), filtered and the filter cake dried to give 5-bromoimidazo [1,2-a ] pyridine-2-carboxylic acid (52) (1.24g, 90% yield, white solid) which was used in the next step.

LCMS:t_R＝0.342min in 5-95AB_1.5min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*30mm,3um),MS(ESI)m/z＝242.9[M+2+H]⁺.

Step 22.3 preparation of Compound 53

5-Bromoimidazol [1,2-a ] pyridine-2-carboxylic acid (27) (1.24g,5.14mmol), 4A molecular sieve (2.48g,10.29mmol) and triethylamine (1.56g,15.43mmol) were suspended in t-butanol (12mL), and diphenyl phosphorazidate (2.12g,7.72mmol) was added. The reaction solution was stirred at 90 ℃ for 16 hours under a nitrogen atmosphere. The reaction was filtered, the filtrate concentrated in vacuo, and the crude product purified by silica gel chromatography (methanol/dichloromethane, from 0% to 2%) to give tert-butyl 5-bromoimidazo [1,2-a ] pyridine-2-carbamate (53) (1.01g, 49% yield, 78% purity, yellow solid).

LCMS:t_R＝1.443min in 10-80AB_2min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*3mm,3um),MS(ESI)m/z＝312.1[M+H]⁺.

Step 22.4 preparation of Compound 54

Tert-butyl 5-bromoimidazo [1,2-a ] pyridine-2-carbamate (53) (1.01g,3.24mmol) was dissolved in dichloromethane (20mL), trifluoroacetic acid (12.32g,108.05mmol) was added, and the reaction mixture was stirred at 15 ℃ for 2 hours. The reaction solution was concentrated, adjusted to pH 9 with saturated sodium bicarbonate, and then extracted with dichloromethane (30mL × 3). The organic phases were combined, washed with brine (20mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product, which was purified by silica gel chromatography (methanol/dichloromethane, from 0% to 5%) to give 5-bromoimidazo [1,2-a ] pyridin-2-amine (54) (400mg, 58% yield, yellow solid).

1H NMR(400MHz,DMSO-d₆):δ＝7.24(d,J＝8.0Hz,1H),7.06(d,J＝7.2Hz,1H),7.02-6.97(m,2H),5.30(brs,2H).

Step 22.5 preparation of Compound 46

5-Bromoimidazo [1,2-a ] pyridin-2-amine (54) (400mg,1.89mmol) was dissolved in N, N-dimethylacetamide (2mL) and cyclopropylcarbonyl chloride (237mg,2.26mmol) was added and stirred at 15 ℃ for 2 hours. The reaction was quenched by addition of saturated sodium bicarbonate (20mL), filtered, the filter cake washed with water (10mL), and dried to give N- (5-bromoimidazo [1,2-a ] pyridin-2-yl) cyclopropylcarboxamide (46) (460mg, 87% yield, yellow solid).

LCMS:t_R＝1.263min in 5-95AB_2min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*3mm,3um),MS(ESI)m/z＝280.0[M+H]⁺.

Step 22.6 preparation of Compound EXP-51

Reacting [5- (methanesulfonamido) benzothiophen-2-yl]Boronic acid (3) (242mg,0.892mmol), N- (5-bromoimidazo [1, 2-a)]Pyridin-2-yl) cyclopropylcarboxamide (46) (100mg,0.357mmol) and potassium carbonate were suspended in dioxane (2mL) and water (0.4mL) and replaced three times with nitrogen, then 1, 1-bis (diphenylphosphino) ferrocene palladium chloride (39mg,0.0535mmol) was added and replaced three times with nitrogen, stirring at 100 ℃ for 2 hours. The reaction was concentrated in vacuo to give crude which was purified by silica gel chromatography (methanol/dichloromethane, from 0% to 4%) and further subjected to preparative HPLC (Column: AgelaDuraShell C18150: 25mm 5 um; Condition: water (0.04% NH)_3.H₂O+10mM NH₄HCO₃) -ACN; b30% for Begin, and End B60%; gradient Time (8 min); FlowRate (25ml/min)) was purified to give N- [5- [5- (methanesulfonamido) benzothien-2-yl ] -N- [ 3- (methyl-phenyl-amino) benzothien-2-yl ] -amide]Imidazole [1,2-a ]]Pyridin-2-yl]Cyclopropylcarboxamide (EXP-51) (10.2mg, 6% yield, 94.5% purity, yellow solid).

LCMS:t_R＝3.255min in 10-80AB_7min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*3mm,3um),MS(ESI)m/z＝427.2[M+H]⁺.

HPLC:t_R＝2.75min in 10-80AB_8min.met.

1H NMR(400MHz,DMSO-d₆):δ＝11.12(s,1H),9.90(br s,1H),8.37(s,1H),8.09-8.01(m,2H),7.84(d,J＝2.0Hz,1H),7.55(d,J＝8.8Hz,1H),7.41-7.31(m,2H),7.18(d,J＝7.2Hz,1H),3.02(s,3H),1.98-1.88(m,1H),0.85-0.74(m,4H).

The example compounds of the following table 10 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 10 ]

Example 23: detection of JAK kinase inhibitory Activity

The compounds of the above examples were applied to the detection and screening of JAK kinase inhibitory activity.

1. Method of producing a composite material

The first 96-well plate was loaded with the corresponding volume of DMSO per well. Each well is filled with 20-30 μ L of 100-200 μ M stock solutions of different test compounds. Mix for 3min with shaking. All compounds were diluted 3X serially. Adding 60-80 mu L of Kinase buffer into each hole of the second 96-hole plate, and taking 1-2 mu L of solution from each hole of the first 96-hole plate and adding the solution into the corresponding hole of the second 96-hole plate. Mix for 3min with shaking. From the second compound dilution plate, 5. mu.L of compound dilution per well was transferred to the corresponding well of the 384-well test plate. 1-2 μ L TK Substrate-biotin was added to each well of the test plate. Adding 1-2 mu L of enzyme mixed solution into each hole. Blank wells were also provided with no enzyme added. Adding 1-2 mu L of ATP solution into each hole, sealing the plate, and reacting at room temperature. The reaction time is respectively as follows: 2 hours JAK1, 30min JAK2, 30min JAK3 and 50min TYK 2. Adding 3-5 mu Lof Streptavidin-XL665 and 3-5 mu L of TK Antibody-cryptate into each well, sealing the plate, and standing at room temperature for 30 minutes to finish the reaction. Fluorescences at 665nm and 620nm were read on a Perkinelmer EnVision machine.

IC50 calculation

Calculate the HTRF ratio for each well: (665signal/620signal) x 10^4.

Percent inhibition was calculated based on the following equation:

inhibition% (% Ratio)_max-Ratio_{Compound (I)})/(Ratio_max-Ratio_min)]×100％

Wherein Ratio_{Compound (I)}Is the HTRF Ratio, Ratio at a given compound concentration_minHTRFratio, Ratio for addition to blank well_maxIs the HTRF ratio without the addition of a compound. Sigmoidal dose-inhibition curves were plotted using a non-linear regression model and IC was calculated by using GraphPad Prism 5.0 software₅₀The value is obtained.

Filgotinib represents a compound having the following structure:

2. results of the experiment

IC measured for each test compound₅₀The (nM) measurements are shown in Table 11 below.

[ TABLE 11 ] inhibitory Activity of Compounds on JAK

Remarking: IC (integrated circuit)₅₀Values from 0 to 10nM are marked A; 10-30nM is marked B; 30-50nM is marked C; the 50-500nM marker is D; greater than 500nM is labeled E; NT stands for not tested.

Claims

1. A compound of formula (I) or a stereoisomer, tautomer, or pharmaceutically acceptable salt or solvate or prodrug thereof:

wherein:

a is independently selected from S, N, N (Me);

d is independently selected from C, N;

R₃independently selected from H, substituted or unsubstituted: sulfonyl, sulfonamide, 5-to 6-membered heterocycle having at least one heteroatom and optionallyIndependently of a second ring heteroatom selected from N and S, substituted or unsubstituted C₁-C₆Alkane, the substituents being independently selected from 5 to 6 membered heteroaryl and methoxy substituted, substituted or unsubstituted C₃-C₇Cycloalkyl, substituted or unsubstituted 4-7 membered heterocycloalkyl, substituted or unsubstituted 5-7 membered heteroaryl.

m is 0, 1,2, 3; n is 0, 1,2, 3; t is 0, 1,2, 3.

2. A compound of formula (I) according to claim 1, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt or solvate or prodrug thereof, characterized in that B is selected from the group consisting of substituted or unsubstituted: cyclopropane, cyclobutane, cyclopentane, cyclohexane, pyrazolyl, pyridyl, thiophene, thiazole, 1,3, 4-thiadiazole, oxazole, 1,3, 4-oxadiazole, furan, pyrrole, 3-pyrroline, 2-pyrazoline, imidazole, 1,2, 3-triazole, 1,2, 4-triazole, pyran, pyridazine, pyrimidine, pyrazine; the substituent is selected from halogen and C₁-C₃Alkyl radical, C₁-C₃A haloalkyl group; preferably, B is selected from the following substituted or unsubstituted groups: cyclopropane, cyclobutane, pyrazolyl, pyridyl, imidazolyl; the substituent is selected from F, Cl, Br, methyl, ethyl, propyl, isopropyl and trifluoromethyl.

3. The compound of formula (I) according to claim 1, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt or solvate or prodrug thereof, wherein D is C or N; n is 1; m is 0 or 1, preferably, m is 0; t is 1.

4. The compound of formula (I) according to claim 1, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt or solvate or prodrug thereof, wherein R is₁Independently selected from H, -NH₂-COOH, hydroxy, halogen, substituted or unsubstituted C₁-C₃Alkyl, substituted or unsubstituted C₁-C₃Acyl, substituted or unsubstituted C₁-C₃Acylamino, substituted or unsubstituted C₁-C₃Alkoxy, substituted or unsubstituted C₁-C₃An ester group; preferably, R₁Independently selected from H, -NH₂-COOH, hydroxy, halogen, methyl, ethyl, propyl, isopropyl, formylamino, carbomethoxy; l is₁Is absent or independently selected from the group consisting of a single bond, -CH₂-，-(CH₂)_xO(CH₂)_y-,-(CH₂)_xNH(CH₂)_y-,-CONH(CH₂)_y-,-N(R₄)-；R₄Independently selected from H, methyl, ethyl, propyl, ethyl methyl ether, methyl ether, ethyl ether; x or y are independently selected from 0, 1 or 2; preferably, L₁Is absent or independently selected from the group consisting of a single bond, -CH₂-，-(CH₂)_xO(CH₂)_y-,-(CH₂)_xNH(CH₂)_y-,-CONH(CH₂)_y-,-N(R₄)-；R₄Independently selected from H, methyl, ethyl methyl ether; x or y is independently selected from 0, 1 or 2.

5. The compound of formula (I) according to claim 1, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt or solvate or prodrug thereof, wherein R is₃Independently selected from H, substituted or unsubstituted: sulfonyl, sulfonamide, thiomorpholine 1, 1-dioxide, piperidine, pyrazole, thiazole, imidazole, 1,3, 4-thiadiazole, piperazine, morpholine, pyrimidine, pyrazine, thiophene, oxazole, 1,3, 4-oxadiazole, furan, pyrrole, 3-pyrroline, 2-pyrazoline, 1,2, 3-oxazole, 1,2, 3-triazole, 1,2, 4-triazole, pyran; the substituent is selected from H, halogen and C₁-C₃Alkyl radical, C₁-C₃Haloalkyl, hydroxy C₁-C₃Alkyl radical, C₃-C₆Cycloalkyl, ethyl methyl ether, methyl ether, ethyl ether; preferably, R₃Independently selected from H, substituted or unsubstituted: sulfonyl, sulfonamido, thiomorpholine 1, 1-dioxide, piperidine, pyrazole, thiazole, imidazole, 1,3, 4-thiaOxadiazole, piperazine, morpholine, pyrimidine, thiophene, oxazole, 1,3, 4-oxadiazole, furan, pyrrole, 3-pyrroline, 2-pyrazoline, 1,2, 3-oxazole, 1,2, 3-triazole, 1,2, 4-triazole; the substituent is selected from H, halogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, hydroxyethyl, hydroxymethyl, cyclopropane, cyclobutane, ethyl methyl ether, methyl ether, ethyl ether.

6. A compound represented by the following structure or a stereoisomer, tautomer, or pharmaceutically acceptable salt or solvate or prodrug thereof:

7. a pharmaceutical composition comprising a compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent or carrier.

8. Use of a compound according to any one of claims 1 to 6 for the manufacture of a medicament for the prevention or treatment of a JAK kinase associated disease, preferably for the manufacture of a medicament for the prevention and/or treatment of diseases involving cartilage degradation, bone and/or joint degradation, conditions involving inflammation or immune response, endotoxin driven disease states, cancer and organ transplant rejection; more preferably, the use in the manufacture of a medicament for the prevention and/or treatment of osteoarthritis, crohn's disease, rheumatoid arthritis, psoriasis, allergic airway disease, juvenile idiopathic arthritis, colitis, inflammatory bowel disease, endomycin driven disease states, diseases with impairment of cartilage turnover, congenital cartilage malformations, organ transplant rejection, diseases involving cartilage degradation, joint degradation, myeloproliferative disorders, leukemia, solid tumors.