CN113999205A

CN113999205A - Pyridine compound containing triazole ketoamide and imidazole amide structures and application thereof

Info

Publication number: CN113999205A
Application number: CN202111513719.1A
Authority: CN
Inventors: 刘举; 陈烨; 丁实; 李春艳; 沈继伟; 刘放; 田璐; 温迪; 吴爽; 李振; 张荠芳; 赵乐妍
Original assignee: Liaoning University
Current assignee: Liaoning University
Priority date: 2021-12-13
Filing date: 2021-12-13
Publication date: 2022-02-01
Anticipated expiration: 2041-12-13
Also published as: CN113999205B

Abstract

The invention relates to a pyridine compound containing triazole ketoamide and imidazole amide structures and application thereof. The pyridine compound containing triazole ketoamide and imidazole amide structures has a structure shown in a general formula (I), has a strong effect of inhibiting c-Met kinase, and also relates to application of the compound and pharmaceutically acceptable salts thereof in preparing medicines for treating and/or preventing diseases caused by abnormal high expression of c-Met kinase, in particular to application in preparing medicines for treating and/or preventing cancers.

Description

Pyridine compound containing triazole ketoamide and imidazole amide structures and application thereof

Technical Field

The invention relates to pyridine compounds containing triazole ketoamide and imidazole amide structures shown in a general formula (I) and pharmaceutically acceptable salts thereof, a preparation method thereof and a pharmaceutical composition containing the compounds. The invention also relates to application of the compound and pharmaceutically acceptable salts thereof in preparing medicaments for treating diseases caused by abnormal high expression of c-Met kinase, in particular to application in preparing medicaments for treating and/or preventing cancers.

Background

Cancer generally refers to all malignant tumors, cancer has always been a great threat to human health, and the statistical data of the world health organization shows that in 2020, 1000 million people die of cancer worldwide, and more heartily about 40 million children are diagnosed with cancer every year. One in five people suffer from cancer throughout their lifetime. The cancer mortality rate even exceeds that of cardiovascular and cerebrovascular diseases, and is the first of all diseases. 2140 million new cases and over 1300 million dead people are expected to occur in 2030 s all the world, so that the research on the anti-tumor drugs, in particular the high-efficiency and low-toxicity molecular targeting anti-tumor drugs, has great significance. In recent years, the molecular targeted therapy of tumors is receiving more and more attention. Compared with the traditional tumor treatment method, the molecular targeted therapy can simultaneously achieve the advantages of orientation and positioning, reduce the dosage of the drug, reduce the toxic and side effects and improve the curative effect. Studies have shown that nearly 80% of oncogenes contain tyrosine kinase codes. Inhibition of tyrosine kinase receptors can effectively control the phosphorylation of downstream signals, thereby inhibiting the growth of tumor cells. Therefore, the development of new drugs with tyrosine kinase as a target becomes a long-standing research subject of anti-tumor drugs at home and abroad. c-Met is a prototypic member of the Ron subfamily of the tyrosine kinase receptor family, expressed by the proto-oncogene c-Met, which is the only known Hepatocyte Growth Factor (HGF) high affinity receptor. c-Met is a receptor-type tyrosine kinase, which is expressed in both normal and tumor cells. Under normal physiological conditions, HGF, when bound to c-Met kinase, can cause a range of biological effects, such as: promoting the separation of epithelial cells, promoting the mitosis of placenta, regulating the development and structure formation of lung, kidney and mammary gland, etc. However, the abnormal HGF/c-Met activity is closely related to the occurrence, division, angiogenesis, invasiveness, metastasis, drug resistance, etc. of tumors, and they exhibit abnormally high expression in various tumor tissues, such as lung cancer, breast cancer, colon cancer, prostate cancer, pancreatic cancer, ovarian cancer, etc. The receptor tyrosine kinase c-Met is highly expressed and abnormally activated in a large proportion of cancers. The c-Met kinase is a key node protein in a tumor signal network pathway, is concerned about the interaction with other kinases and receptors on the cell surface, and has become a leading hot spot of research in the medical field. Cabozantinib (Cabozantinib) with a 4-phenoxyquinoline parent nucleus is the first approved small molecule c-Met kinase inhibitor to market, and the U.S. FDA approved Cabozantinib in 2012 for the treatment of progressive, metastatic Medullary Thyroid Cancer (MTC) patients; cabozantinib was approved in 2016 for the treatment of advanced Renal Cell Carcinoma (RCC) patients treated with anti-angiogenesis therapy; the FDA approved cabozantinib for first-line treatment of middle-and high-risk advanced renal cancer patients in 2017; cabozantinib was approved in 2018 for the treatment of advanced hepatocellular carcinoma patients who progressed after receiving sorafenib or other standard systemic treatment. Scientific research finds that the pyridine ring is used for replacing the quinoline structure of cabozantinib, and the obtained 4-phenoxypyridine compound keeps good c-Met kinase activity, such as Altiratinib and Golvatinib, and is in clinical research stages at present. However, the following problems mainly exist in the study of c-Met inhibitors: the clinical application has large toxic and side effects, unsatisfactory clinical treatment effect and pharmacokinetic parameters, low oral bioavailability and the like. Therefore, the development of a novel c-Met kinase inhibitor with novel structure, safety and effectiveness is still the key field of research on antitumor drugs at home and abroad.

Disclosure of Invention

The invention aims to design and synthesize a series of novel pyridine compounds containing triazole ketoamide and imidazole amide structures. In vitro activity screening shows that the compounds have antitumor activity.

The invention provides a pyridine compound containing triazole ketoamide and imidazole amide structures and a pharmaceutically acceptable salt thereof, which are shown in a general formula (I),

wherein:

r is selected from hydrogen and C₁-C₁₀Alkyl, or C₃-C₇A cycloalkyl group;

x is selected from hydrogen or halogen;

l is selected from

R₁Selected from hydrogen, C₁-C₆Alkyl, or C₃-C₆A cycloalkyl group;

ar is selected from C₆-C₁₀Aryl or 5-10 membered heteroaryl; wherein the heteroaryl contains 1-3 heteroatoms selected from N, O or S, and Ar is substituted with 1-3R, which may be the same or different₂Substitution;

R₂selected from hydrogen, hydroxyl, halogen, nitro, ester group, amino, cyano, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, C halogenated by 1-3 halogens₁-C₆Alkyl, C halogenated by 1-3 halogens₁-C₆An alkoxy group.

Further, the pyridine compound containing triazole ketone amide and imidazole amide structures and pharmaceutically acceptable salts thereof,

r is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl;

x is selected from hydrogen, fluorine or chlorine;

l is selected from

R₁Selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclobutyl or cyclopentyl;

ar is selected from phenyl or 5-6 membered heteroaryl; wherein the heteroaryl contains 1-3 heteroatoms selected from N, O or S, and Ar is substituted with 1-3R, which may be the same or different₂Substitution;

R₂selected from hydrogen, hydroxy, halogen, nitro, ester group, amino, cyano, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, trifluoromethyl, monofluoromethoxy, difluoromethoxy, trifluoromethoxy.

Furthermore, the pyridine compound containing the triazole ketone amide and imidazole amide structures and pharmaceutically acceptable salts thereof,

r is selected from methyl, ethyl or cyclopropyl;

x is selected from hydrogen or fluorine;

l is selected from

R₁Selected from hydrogen, methyl, ethyl, or cyclopropyl;

ar is phenyl and Ar is substituted by 1 to 3 identical or different R₂Substitution;

R₂selected from hydrogen, fluoro, chloro, bromo, methyl, methoxy, trifluoromethyl, difluoromethoxy or trifluoromethoxy.

Furthermore, the pyridine compounds containing triazole ketone amide and imidazole amide structures and the pharmaceutically acceptable salts thereof have the following structural formula, but the compounds do not mean any limitation to the invention:

a pharmaceutical composition comprises the pyridine compound containing triazole ketoamide and imidazole amide structures and pharmaceutically acceptable salts thereof as active ingredients and pharmaceutically acceptable excipients.

Furthermore, the pyridine compounds containing triazole ketoamide and imidazole amide structures and configurational isomers shown in the general formula (I) of the present invention can form pharmaceutically acceptable salts with acids according to some common methods in the field of the present invention. Pharmaceutically acceptable addition salts include inorganic and organic acid addition salts, with the following acids being particularly preferred: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, succinic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, and the like.

In addition, the present invention also includes prodrugs of the compounds of the present invention. Prodrugs of the compounds of the present invention are pyridines containing triazolinone amide and imidazolamide structures of formula (i) which may themselves be less active or even inactive, but which upon administration are converted under physiological conditions (e.g. by metabolism, solvolysis or otherwise) to the corresponding biologically active form.

"halogen" in the present invention means fluoro, chloro, bromo or iodo; "alkyl" refers to straight or branched chain alkyl; "alkylene" refers to straight or branched chain alkylene; "cycloalkyl" refers to a substituted or unsubstituted cycloalkyl; "aryl" refers to phenyl, naphthyl with no substituent or with a substituent attached; "heteroaryl" means a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, which ring system is aromatic, such as imidazolyl, pyridyl, pyrazolyl, (1,2,3) -and (1,2,4) -triazolyl, furyl, thienyl, pyrrolyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl, naphthyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl and the like.

The invention has the beneficial effects that:

according to the invention, the c-Met enzyme activity test shows that the compound has significant c-Met kinase inhibition activity, and can be used for preparing c-Met inhibitor drugs, especially drugs for treating and/or preventing cancers, and more especially drugs for treating and/or preventing gastric cancer, lung cancer and colon cancer.

According to the invention, the in vitro inhibition test of the c-Met high expression cell strain human gastric cancer cell strain MKN45, the human lung adenocarcinoma cell strain A549, the human lung cancer cell H460 and the human colon cancer cell HT-29 proves that the compound has a significant inhibition effect on human gastric cancer cells, human lung cancer cells and human colon cancer cells, and is particularly used for preparing medicines for treating and/or preventing lung cancer and colon cancer.

Detailed Description

The examples and preparations provided below further illustrate and exemplify the compounds of the present invention and their methods of preparation. It should be understood that the scope of the following examples and preparations are not intended to limit the scope of the invention in any way. The following examples are intended to illustrate, but not limit, the scope of the invention. NMR of the compounds was measured using Bruker ARX-600 or Bruker ARX-400 and Mass Spectroscopy was measured using Agilent 6460 QQQ; all reagents used were analytically or chemically pure.

The following synthetic scheme (scheme 1) describes the preparation of the compounds of general formula (i) according to the invention, all starting materials being prepared by the means described in these synthetic schemes, by methods well known to the person skilled in the art of organic chemistry or being commercially available. All of the final compounds of the present invention are prepared by the methods described in these synthetic routes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All variables used in these synthetic routes are as defined below or in the claims.

The compounds of general formula (I) according to the invention can be prepared in the manner of scheme 1 from the corresponding intermediates Q and the corresponding M by condensation.

The synthetic route for M is shown in scheme 2, the other substituents are as defined in the claims;

according to the compounds of general formula (I) according to the invention, intermediate Q can be prepared by the process shown in scheme 3, the other substituents being as defined in the claims.

The compounds of general formula (I) according to the invention, intermediates Q, can also be prepared by the process as shown in scheme 4, the other substituents being as defined in the claims.

Substituent R, X, L, R of all intermediates in the above five routes₁Ar is as defined in the claims.

EXAMPLE 1 preparation of N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-fluorophenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 1)

Step 1N- (4-Chloropyridin-2-yl) cyclopropylcarboxamide (a)

2-amino group8.80g of 4-chloropyridine and 20.80g of triethylamine were dissolved in 80mL of methylene chloride, and 30mL of a methylene chloride solution containing 9.30g of cyclopropylcarbonyl chloride was added dropwise to the solution in an ice bath, and the solution was warmed to room temperature after completion of the addition. Stirring for 12h, and reacting, and mixing with 20% K₂CO₃Washing the solution and saturated saline solution for 3 times respectively, separating out an organic phase, drying by anhydrous sodium sulfate, filtering, evaporating the solvent to obtain a crude product, and separating by column chromatography to obtain a white solid N- (4-chloropyridin-2-yl) cyclopropyl formamide (a).

Step 2N- [4- (2-fluoro-4-nitrophenoxy) pyridin-2-yl ] cyclopropylcarboxamide (b)

8.00g of intermediate (a), N- (4-chloropyridin-2-yl) cyclopropylcarboxamide and 15.98g of 2-fluoro-4-nitrophenol were added to 100mL of chlorobenzene and reacted at 140 ℃ for 40 hours. Cooling to room temperature, concentrating under reduced pressure, dissolving the residue in an appropriate amount of dichloromethane, adding₂CO₃Washing the solution and saturated saline solution for 3 times, separating organic phase, drying with anhydrous sodium sulfate, filtering, evaporating solvent to obtain brown solid, and performing column chromatography to obtain light yellow solid product N- [4- (2-fluoro-4-nitrophenoxy) pyridin-2-yl]Cyclopropyl carboxamide (b).¹H NMR(600MHz,DMSO-d₆)δ11.00(s,1H),8.43(m,1H),8.30(d,J＝5.7Hz,1H),8.19(m,1H),7.76(d,J＝2.2Hz,1H),7.61(t,J＝8.5Hz,1H),6.86(m,1H),2.04–1.95(m,1H),0.78(t,J＝6.3Hz,4H)。

Step 3N- [4- (4-amino-2-fluorophenoxy) pyridin-2-yl ] cyclopropylcarboxamide (M)

Intermediate (b) N- [4- (2-fluoro-4-nitrophenoxy) pyridin-2-yl]6.00g of cyclopropyl formamide, 5.28g of iron powder and 11.36g of acetic acid are added into 100mL of ethyl acetate, 20mL of water is added, heating reflux is carried out for 2h, the reaction is finished, the hot reaction product is filtered, an organic phase is separated, anhydrous sodium sulfate is dried, the filtration is carried out, and the solvent is evaporated to dryness under reduced pressure to obtain a white solid N- [4- (4-amino-2-fluorophenoxy) pyridin-2-yl]Cyclopropyl carboxamide (M).¹H NMR(600MHz,DMSO-d₆)δ10.79(s,1H),8.15(d,J＝5.7Hz,1H),7.59(s,1H),6.95(t,J＝9.0Hz,1H),6.67–6.61(m,1H),6.49(dd,J＝13.1,2.2Hz,1H),6.40(d,J＝8.7Hz,1H),5.44(s,2H),2.03–1.88(m,1H),0.76(br,4H)。

Step 42-chloro-2- (2-fluorobenzylhydrazone) acetic acid ethyl ester (c)

Taking 7.11g of o-fluoroaniline, adding 20mL of ethanol, 7mL of water and 13mL of concentrated hydrochloric acid, dissolving, cooling the solution to below-5 ℃, dropwise adding 5.77g of sodium nitrite solution dissolved in 15mL of water, keeping the temperature below 5 ℃ in the dropwise adding process, and stirring for 0.5h after dropwise adding. The temperature is reduced to below 0 ℃, 15.6g of anhydrous sodium acetate dissolved by 90mL of water is dripped, and 10.53g of 2-chloroacetoacetic acid ethyl ester is added after dripping. Stirring for 5h at room temperature, filtering, pulping filter cake with cold ethanol, filtering, and drying to obtain light yellow product ethyl 2-chloro-2- (2-fluorobenzene hydrazone) acetate (c)12.4g, yield 79%.¹H NMR(600MHz,CDCl₃)δ8.49(br,1H),7.63(m,1H),7.16(t,J＝7.8Hz,1H),7.11(m,1H),6.99(m,1H),4.41(q,J＝7.2Hz,2H),1.43(t,J＝7.2Hz,3H),；MS(ESI)m/z(％):267.1[M+Na]⁺.

Step 52-methylamino-2- (2-fluorobenzylhydrazone) acetic acid ethyl ester (d)

And (3) dissolving 8.54g of the intermediate (c) ethyl 2-chloro-2- (2-fluorobenzylhydrazone) acetate in 50mL of tetrahydrofuran, dropwise adding 8.3g of 40% methylamine water solution in an ice bath, keeping the temperature and stirring for 5h, evaporating to remove THF, adding 200mL of water, separating out a solid, filtering, and drying to obtain 7.1g of a yellow product, namely ethyl 2-methylamino-2- (2-fluorobenzylhydrazone) acetate (d), wherein the yield is 85%.¹H NMR(400MHz,CDCl₃)δ10.38(br,1H),7.48(m,1H),7.15–6.90(m,2H),6.88–6.64(m,1H),4.37(m,3H),2.86(s,3H),1.39(t,J＝7.2Hz,3H)；MS(ESI)m/z(％):240.2[M+H].。

Step 64-methyl-5-oxo-1- (2-methylphenyl) -4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxylic acid ethyl ester (e)

Taking 4.78g of intermediate (d), ethyl 2-methylamino-2- (2-fluorobenzene hydrazone) acetate and 6.0g of potassium carbonate, adding 20mL of dioxane, dropwise adding 6.0g of p-nitrophenylchloroformate dissolved in 20mL of dioxane under ice bath, reacting for 2h after the dropwise adding is finished, continuing to perform reflux reaction for 6h, performing suction filtration, and washing and discarding filter cakes of dioxane. The filtrate was evaporated to dryness, dissolved in dichloromethane, washed three times with saturated sodium bicarbonate solution, once with saturated sodium chloride solution, dried over anhydrous sodium sulfate of the organic phase and evaporated to dryness. Column chromatography separation to obtain offwhite product 4-methyl-5-oxo-1- (2-fluorophenyl) -4, 5-dihydro-1H-1, 2, 4-triazole-3-ethyl formateEster (e)2.8g, 53% yield.¹H NMR(400MHz,CDCl₃)δ7.55(m,2H),7.44(m,1H),7.33(m,1H),4.35(q,J＝7.2Hz,2H),3.45(s,3H),1.30(t,J＝7.2Hz,3H)；MS(ESI)m/z(％):288.1[M+Na]⁺。

Step 74-methyl-5-oxo-1- (2-fluorophenyl) -4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxylic acid (Q-1)

And (3) dissolving 2.1g of the intermediate (e), namely 4-methyl-5-oxo-1- (2-fluorophenyl) -4, 5-dihydro-1H-1, 2, 4-triazole-3-ethyl formate in 32mL of tetrahydrofuran, adding 8mL of water, adding 640mg of lithium hydroxide monohydrate while stirring, stirring at room temperature for reaction, and finishing the reaction for 60 min. THF is removed by reduced pressure evaporation, 30mL of water is added, ethyl acetate is used for extraction twice, the pH value of a water phase is adjusted by 6mol/L hydrochloric acid, a white solid is separated out, the filtration is carried out, a filter cake is washed by water and dried, and 1.4g of a product 4-methyl-5-oxo-1- (2-fluorophenyl) -4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxylic acid (Q-1) is obtained, wherein the yield is 75%.¹H NMR(400MHz,CD₃OD)δ7.57(m,2H),7.35(m,2H),3.62(s,3H)；MS(ESI)m/z(％):236.2[M-H]^—.

Step 8N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-fluorophenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 1)

Adding the intermediate (Q-1) 4-methyl-5-oxo-1- (2-fluorophenyl) -4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxylic acid (1.20mmol) and the intermediate (M) N- [4- (4-amino-2-fluorophenoxy) pyridin-2-yl into a reaction bottle]Cyclopropylformamide (1.00mmol), HATU (1.20mmol), triethylamine (1.20mmol), 15mL DMF, and stirred at room temperature overnight to complete the reaction. The reaction solution was poured into 100mL of 20% aqueous sodium carbonate solution, extracted three times with 50mL of dichloromethane, the organic phases were combined, the organic phase was washed three times with 20% aqueous sodium carbonate solution, the organic layer was washed twice with saturated brine, and the separated organic layer was dried over anhydrous sodium sulfate. Filtering, distilling off dichloromethane under reduced pressure, and separating by column chromatography to obtain N- (4- { [2- (cyclopropane carboxamido) pyridine-4-yl]Oxy } -3-fluorophenyl) -1- (2-fluorophenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (compound 1).¹H NMR(600MHz,DMSO-d₆)δ11.02(s,1H),10.88(s,1H),8.21(d,J＝5.7Hz,1H),7.94(dd,J＝12.9,2.3Hz,1H),7.72(dd,J＝8.9,1.3Hz,1H),7.69–7.61(m,2H),7.61–7.54(m,1H),7.54–7.45(m,1H),7.44–7.32(m,2H),6.74(dd,J＝5.7,2.4Hz,1H),3.54(s,3H),1.97(dt,J＝12.5,6.3Hz,1H),0.77(d,J＝6.1Hz,4H)；MS(ESI)m/z(％):507.20[M+H]⁺,529.20[M+Na]⁺。

Examples 2 to 13 (compounds 2 to 13) were finally obtained according to the preparation scheme of example 1, replacing the appropriate starting materials and reagents.

EXAMPLE 2N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-bromophenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 2)

¹H NMR(600MHz,DMSO-d₆)δ11.05(s,1H),10.89(s,1H),8.22(d,J＝5.5Hz,1H),7.95(d,J＝12.8Hz,1H),7.88(d,J＝7.9Hz,1H),7.73(d,J＝8.6Hz,1H),7.70–7.56(m,3H),7.52(t,J＝7.6Hz,1H),7.40(t,J＝8.9Hz,1H),6.74(d,J＝4.8Hz,1H),3.56(s,3H),2.13–1.86(m,1H),0.77(d,J＝5.7Hz,4H)。

EXAMPLE 3N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-methoxyphenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 3)

¹H NMR(600MHz,DMSO-d₆)δ10.97(s,1H),10.89(s,1H),8.22(d,J＝5.5Hz,1H),7.94(d,J＝12.5Hz,1H),7.72(d,J＝8.5Hz,1H),7.64(s,1H),7.53(t,J＝7.6Hz,1H),7.48–7.33(m,2H),7.25(d,J＝8.2Hz,1H),7.10(t,J＝7.4Hz,1H),6.74(d,J＝3.5Hz,1H),3.81(s,3H),3.53(s,3H),2.02–1.92(m,1H),0.77(d,J＝5.7Hz,4H)；MS(ESI)m/z(％):519.2[M+H]⁺。

EXAMPLE 4N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (3-fluorophenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 4)

¹H NMR(600MHz,DMSO-d₆)δ10.93(s,1H),10.88(s,1H),8.25–8.19(m,1H),7.99–7.87(m,3H),7.72(dd,J＝8.8,1.4Hz,1H),7.65(d,J＝2.3Hz,1H),7.62–7.56(m,1H),7.43(t,J＝9.0Hz,1H),7.18(td,J＝8.4,2.0Hz,1H),6.75(dd,J＝5.7,2.4Hz,1H),3.53(s,3H),2.00–1.93(m,1H),0.80–0.72(m,4H)；MS(ESI)m/z(％):507.2[M+H]⁺。

EXAMPLE 5N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-chlorophenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 5)

¹H NMR(600MHz,DMSO-d₆)δ10.99(s,1H),10.89(s,1H),8.23(d,J＝5.7Hz,1H),8.09(d,J＝8.9Hz,2H),7.95(dd,J＝12.8,2.2Hz,1H),7.73(d,J＝8.8Hz,1H),7.64(dd,J＝12.4,5.5Hz,3H),7.44(t,J＝9.0Hz,1H),6.76(dd,J＝5.7,2.3Hz,1H),3.53(s,3H),2.03–1.93(m,1H),0.80–0.74(m,4H)。

EXAMPLE 6N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-difluoromethoxyphenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 6)

IR(KBr,cm^-1):3739.9,1710.9,1516.1,1427.3,1211.3,1122.6,1041.6,831.3；¹H NMR(600MHz,DMSO-d₆)δ10.97(s,1H),10.88(s,1H),8.22(d,J＝5.7Hz,1H),8.10–8.03(m,2H),7.94(dd,J＝12.8,2.4Hz,1H),7.72(dd,J＝8.9,1.3Hz,1H),7.64(d,J＝2.3Hz,1H),7.43–7.39(m,1H),7.37(d,J＝9.0Hz,2H),7.28(t,J＝73.9Hz,1H),6.75(dd,J＝5.7,2.4Hz,1H),3.53(s,3H),2.01–1.92(m,1H),0.79–0.74(m,4H)；MS(ESI)m/z(％):555.2[M+H]⁺。

EXAMPLE 7N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-bromophenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 7)

MS(ESI)m/z(％):567.2[M+H]⁺。

EXAMPLE 8N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-trifluoromethoxyphenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 8)

MS(ESI)m/z(％):573.2[M+H]⁺。

EXAMPLE 9N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2, 4-dichlorophenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 9)

IR(KBr,cm^-1):3614.6,3242.3,2370.5,2308.8,1726.3,1602.9,1516.1,1419.6,1303.9,1192.0,1111.0,987.6,827.5,723.3；¹H NMR(600MHz,DMSO-d₆)δ11.11(s,1H),10.93(s,1H),8.26(d,J＝5.7Hz,1H),7.99(d,J＝2.1Hz,2H),7.77(d,J＝8.6Hz,2H),7.73–7.63(m,2H),7.44(t,J＝9.0Hz,1H),6.79(dd,J＝5.6,2.2Hz,1H),3.60(s,3H),2.07–1.97(m,1H),0.82(d,J＝6.0Hz,4H)；MS(ESI)m/z(％):557.1[M+H]⁺,579.1[M+Na]⁺。

EXAMPLE 10N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2, 6-dimethylphenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 10)

¹H NMR(600MHz,DMSO-d₆)δ10.92(s,1H),10.88(s,1H),8.21(d,J＝5.7Hz,1H),7.95(dd,J＝12.9,2.4Hz,1H),7.73(dd,J＝8.9,1.3Hz,1H),7.63(d,J＝2.3Hz,1H),7.50–7.31(m,2H),7.24(d,J＝7.6Hz,2H),6.74(dd,J＝5.7,2.4Hz,1H),3.57(s,3H),2.13(s,6H),2.01–1.91(m,1H),0.82–0.72(m,4H)；MS(ESI)m/z(％):517.2[M+H]⁺,539.2[M+Na]⁺。

EXAMPLE 11N- (4- { [2- (acetylamino) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-fluorophenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 11)

MS(ESI)m/z(％):481.2[M+H]⁺。

EXAMPLE 12N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-fluorophenyl) -4-cyclopropyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 12)

MS(ESI)m/z(％):533.2[M+H]⁺,555.2[M+Na]⁺。

EXAMPLE 13N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } phenyl) -1- (2-fluorophenyl) -4-methyl-5-oxo-4, 5-dihydro-1H-1, 2, 4-triazole-3-carboxamide (Compound 13)

IR(KBr,cm^-1):3612.7,3250.1,1712.8,1604.8,1527.6,1302.0,1203.6,995.3,839.0,754.2；¹H NMR(600MHz,DMSO-d₆)δ10.85(s,1H),10.83(s,1H),8.19(d,J＝5.7Hz,1H),7.87(d,J＝8.9Hz,2H),7.72–7.62(m,2H),7.61–7.53(m,1H),7.49(t,J＝9.3Hz,1H),7.40(t,J＝7.7Hz,1H),7.18(d,J＝8.9Hz,2H),6.69(dd,J＝5.7,2.3Hz,1H),3.54(s,3H),2.01–1.92(m,1H),0.76(d,J＝6.1Hz,4H)；MS(ESI)m/z(％):489.2[M+H]⁺,511.2[M+Na]⁺。

EXAMPLE 14 preparation of N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-fluorophenyl) -1H-imidazole-4-carboxamide (Compound 14)

Step 1N- (2-fluorophenyl) carboxamide (f)

Taking 10.7g of 2-fluoroaniline, 2.08g of sodium formate dihydrate and 50mL of anhydrous formic acid, and carrying out reflux reaction for 16h to finish the reaction. The solvent was removed by evaporation under reduced pressure, and the residue was added to 150mL of ethyl acetate, washed with water three times, dried with the organic phase and concentrated under reduced pressure to give 12.6g of N- (2-fluorophenyl) formamide (f), a white product, in 94% yield. ESI-MS, 140.1[ M-1 ]]^—。

Step 22-Fluorophenylisocyanide (g)

6.96g of intermediate (f), N- (2-fluorophenyl) formamide, was dissolved in 40mL of THF, 20.25g of triethylamine was added thereto, and a mixture of 10.0g of phosphorus oxychloride and 20mL of THF was added dropwise while controlling the reaction temperature to 0 ℃ or lower. Keeping the temperature at 0 ℃ for 3h, heating to room temperature for reaction for 1h, slowly pouring the reaction liquid into ice water, separating out a solid, filtering and drying to obtain a light yellow solid product, namely 4.4g of 2-fluorophenyl isonitrile (g). The yield was 73%.

Step 31- (2-fluorophenyl) -1H-imidazole-4-carboxylic acid ethyl ester (H)

Taking 6.06g of the intermediate (g), 7.92g of ethyl isocyanate, 1.00g of cuprous oxide and 1.98g of phenanthroline monohydrate, adding 30mL of THF for dissolving, and carrying out reflux reaction for 4h to finish the reaction. Evaporation under reduced pressureThe solvent was removed. Ethyl acetate (100 mL) was added under reflux, and the insoluble matter was filtered off while it was still hot. The filtrate was evaporated under reduced pressure to about 15-20mL, cooled to precipitate a solid, filtered, and the filter cake was washed with cold ethyl acetate to give 4.9g of ethyl 1- (2-fluorophenyl) -1H-imidazole-4-carboxylate (H), a white solid product, 42% yield. ESI-MS (M/z):235.1[ M + H]⁺。

Step 41- (2-fluorophenyl) -1H-imidazole-4-carboxylic acid (Q-2)

Taking 2.1g of intermediate (H) ethyl 1- (2-fluorophenyl) -1H-imidazole-4-carboxylate, dissolving in 30mL of tetrahydrofuran, adding 8mL of water, adding 770mg of lithium hydroxide monohydrate with stirring, and carrying out reflux reaction with stirring, wherein the reaction is finished for 4H. THF was removed by evaporation under reduced pressure, 30mL of water was added, extraction was performed twice with ethyl acetate, the aqueous phase was adjusted to pH 2 with 6mol/L hydrochloric acid, a white-like solid was precipitated, filtered, the filter cake was washed with water and dried to give 1.5g of 1- (2-fluorophenyl) -1H-imidazole-4-carboxylic acid (Q-2) as a product in 81% yield. ESI-MS (M/z):205.1[ M-H]^—。

Step 5N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-fluorophenyl) -1H-imidazole-4-carboxamide (Compound 14)

Intermediate (Q-2)1- (2-fluorophenyl) -1H-imidazole-4-carboxylic acid (1.20mmol), intermediate (M) N- [4- (4-amino-2-fluorophenoxy) pyridin-2-yl]Cyclopropylformamide (1.00mmol), HATU (1.20mmol), triethylamine (1.20mmol), 15mL DMF, and stirred at room temperature overnight to complete the reaction. The reaction solution was poured into 100mL of 20% aqueous sodium carbonate solution, extracted three times with 50mL of dichloromethane, the organic phases were combined, the organic phase was washed three times with 20% aqueous sodium carbonate solution, the organic layer was washed twice with saturated brine, and the separated organic layer was dried over anhydrous sodium sulfate. Filtering, distilling off dichloromethane under reduced pressure, and separating by column chromatography to obtain N- (4- { [2- (cyclopropane carboxamido) pyridine-4-yl]Oxy } -3-fluorophenyl) -1- (2-fluorophenyl) -1H-imidazole-4-carboxamide (compound 14). IR (KBr, cm)^-1):3417.6,3353.9,3144.3,3005.5,1799.7,1681.6,1599.5,1581.7,1513.9,1465.0,1436.0,1389.7,1302.1,1268.1,1208.6,1124.9,1112.0,1060.1,1028.8；¹H NMR(600MHz,CDCl₃)δ9.12(s,1H),9.03(s,1H),8.11(d,J＝5.8Hz,1H),8.00(s,1H),7.91(dd,J＝12.1,2.3Hz,1H),7.83(d,J＝14.7Hz,2H),7.45(dd,J＝10.1,5.8Hz,2H),7.39–7.30(m,3H),7.16(t,J＝8.7Hz,1H),6.61(dd,J＝5.8,2.2Hz,1H),1.68–1.53(m,1H),1.16–1.00(m,2H),0.92–0.79(m,2H)；MS(ESI)m/z(％):476.1[M+H]⁺,498.1[M+Na]⁺。

Examples 15 to 33 (compounds 15 to 33) were finally obtained according to the preparation scheme of example 14, replacing the appropriate starting materials and reagents.

EXAMPLE 15N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-chlorophenyl) -1H-imidazole-4-carboxamide (Compound 15)

¹H NMR(600MHz,DMSO-d₆)δ10.87(s,1H),10.42(s,1H),8.30–8.12(m,3H),8.07(d,J＝13.1Hz,1H),7.88–7.63(m,4H),7.58(t,J＝6.7Hz,2H),7.35(t,J＝9.0Hz,1H),6.74(d,J＝5.4Hz,1H),2.10–1.82(m,1H),0.87–0.69(m,4H)；MS(ESI)m/z(％):492.1[M+H]⁺,514.1[M+Na]⁺。

EXAMPLE 16N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-bromophenyl) -1H-imidazole-4-carboxamide (Compound 16)

¹H NMR(600MHz,DMSO)δ10.87(s,1H),10.42(s,1H),8.21(d,J＝5.7Hz,1H),8.15(d,J＝7.1Hz,2H),8.07(d,J＝13.1Hz,1H),7.90(d,J＝8.0Hz,1H),7.81(d,J＝8.8Hz,1H),7.71–7.63(m,2H),7.60(t,J＝7.6Hz,1H),7.51(t,J＝7.7Hz,1H),7.35(t,J＝9.0Hz,1H),6.73(d,J＝5.4Hz,1H),2.14–1.81(m,1H),0.89–0.58(m,4H)；MS(ESI)m/z(％):536.1[M+H]⁺,558.1[M+Na]⁺。

EXAMPLE 17N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-trifluoromethoxyphenyl) -1H-imidazole-4-carboxamide (Compound 17)

¹H NMR(600MHz,CDCl₃)δ9.11(s,1H),8.71(s,1H),8.11(d,J＝5.8Hz,1H),7.96–7.87(m,2H),7.82(d,J＝1.3Hz,1H),7.73(d,J＝0.9Hz,1H),7.61–7.42(m,4H),7.36(d,J＝7.8Hz,1H),7.16(t,J＝8.7Hz,1H),6.60(dd,J＝5.8,2.3Hz,1H),1.63–1.46(m,1H),1.15–0.99(m,2H),0.94–0.77(m,2H)；MS(ESI)m/z(％):542.2[M+H]⁺,564.2[M+Na]⁺。

EXAMPLE 18N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-fluorophenyl) -1H-imidazole-4-carboxamide (Compound 18)

¹H NMR(600MHz,DMSO-d₆)δ10.87(s,1H),10.41(s,1H),8.47(d,J＝2.9Hz,2H),8.21(d,J＝5.7Hz,1H),8.09(dd,J＝13.2,1.9Hz,1H),7.93–7.75(m,3H),7.66(d,J＝1.7Hz,1H),7.43(t,J＝8.7Hz,2H),7.36(t,J＝9.0Hz,1H),6.74(dd,J＝5.7,2.2Hz,1H),2.09–1.81(m,1H),0.87–0.70(m,4H)；MS(ESI)m/z(％):476.1[M+H]⁺,498.1[M+Na]⁺。

EXAMPLE 19N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-chlorophenyl) -1H-imidazole-4-carboxamide (Compound 19)

¹H NMR(600MHz,DMSO-d₆)δ10.87(s,1H),10.42(s,1H),8.52(d,J＝5.1Hz,2H),8.21(d,J＝5.7Hz,1H),8.08(d,J＝13.1Hz,1H),7.95–7.75(m,3H),7.64(d,J＝8.7Hz,3H),7.36(t,J＝9.0Hz,1H),6.74(dd,J＝5.6,2.1Hz,1H),2.12–1.77(m,1H),0.90–0.66(m,4H)；MS(ESI)m/z(％):492.1[M+H]⁺,514.1[M+Na]⁺。

EXAMPLE 20N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-chlorotrifluoromethylphenyl) -1H-imidazole-4-carboxamide (Compound 20)

¹H NMR(600MHz,DMSO-d₆)δ10.88(s,1H),10.46(s,1H),8.65(d,J＝11.7Hz,2H),8.21(d,J＝5.7Hz,1H),8.09(d,J＝8.9Hz,3H),7.95(d,J＝8.3Hz,2H),7.83(d,J＝8.8Hz,1H),7.65(s,1H),7.36(t,J＝9.0Hz,1H),6.86–6.58(m,1H),2.09–1.82(m,1H),0.86–0.62(m,4H)；MS(ESI)m/z(％):526.2[M+H]⁺,548.2[M+Na]⁺。

EXAMPLE 21N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-methoxyphenyl) -1H-imidazole-4-carboxamide (Compound 21)

¹H NMR(600MHz,DMSO-d₆)δ10.87(s,1H),10.38(s,1H),8.39(s,2H),8.21(d,J＝5.7Hz,1H),8.08(dd,J＝13.3,2.4Hz,1H),7.82(dd,J＝8.9,1.3Hz,1H),7.76–7.68(m,2H),7.65(d,J＝2.3Hz,1H),7.35(t,J＝9.1Hz,1H),7.20–7.01(m,2H),6.74(dd,J＝5.7,2.4Hz,1H),3.83(s,3H),2.06–1.84(m,1H),0.89–0.68(m,4H)；MS(ESI)m/z(％):488.2[M+H]⁺,510.2[M+Na]⁺。

EXAMPLE 22N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-methylphenyl) -1H-imidazole-4-carboxamide (Compound 22)

¹H NMR(600MHz,DMSO-d₆)δ10.80(s,1H),10.32(s,1H),8.38(d,J＝7.4Hz,2H),8.14(d,J＝5.7Hz,1H),8.01(dd,J＝13.2,1.9Hz,1H),7.75(d,J＝8.8Hz,1H),7.67–7.52(m,3H),7.39–7.20(m,3H),6.66(dd,J＝5.7,2.2Hz,1H),2.30(s,3H),2.04–1.77(m,1H),0.80–0.61(m,4H)；MS(ESI)m/z(％):472.2[M+H]⁺,494.2[M+Na]⁺。

EXAMPLE 23N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-bromo-4-fluorophenyl) -1H-imidazole-4-carboxamide (Compound 23)

¹H NMR(600MHz,DMSO-d₆)δ10.80(s,1H),10.35(s,1H),8.14(d,J＝5.7Hz,1H),8.09–8.02(m,2H),8.02–7.96(m,1H),7.84(dd,J＝8.3,2.8Hz,1H),7.76–7.71(m,1H),7.68(dd,J＝8.8,5.5Hz,1H),7.58(d,J＝2.2Hz,1H),7.48–7.38(m,1H),7.28(t,J＝9.0Hz,1H),6.66(dd,J＝5.7,2.4Hz,1H),1.97–1.81(m,1H),0.74–0.65(m,4H)；MS(ESI)m/z(％):556.1[M+H]⁺,578.1[M+Na]⁺。

EXAMPLE 24N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-chloro-4-fluorophenyl) -1H-imidazole-4-carboxamide (Compound 24)

MS(ESI)m/z(％):510.1[M+H]⁺,532.1[M+Na]⁺。

EXAMPLE 25N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (3,4, 5-trifluorophenyl) -1H-imidazole-4-carboxamide (Compound 25)

MS(ESI)m/z(％):512.1[M+H]⁺。

EXAMPLE 26N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2, 6-difluorophenyl) -1H-imidazole-4-carboxamide (Compound 26)

¹H NMR(600MHz,CDCl₃)δ9.11(s,1H),8.70(s,1H),8.15(dd,J＝34.8,5.6Hz,1H),7.98–7.86(m,2H),7.82(s,1H),7.72(s,1H),7.50–7.40(m,1H),7.36(d,J＝8.7Hz,1H),7.16(t,J＝8.6Hz,3H),6.60(dd,J＝5.7,2.2Hz,1H),1.63–1.40(m,1H),1.15–0.99(m,2H),0.94–0.76(m,2H)；MS(ESI)m/z(％):494.1[M+H]⁺,516.1[M+Na]⁺。

EXAMPLE 27N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2, 6-dimethylphenyl) -1H-imidazole-4-carboxamide (Compound 27)

¹H NMR(600MHz,CDCl₃)δ9.16(s,1H),8.71(s,1H),8.11(d,J＝5.8Hz,1H),7.91(dd,J＝12.2,2.4Hz,1H),7.82(d,J＝1.3Hz,1H),7.68(d,J＝1.0Hz,1H),7.45(d,J＝1.0Hz,1H),7.37(dd,J＝8.7,1.1Hz,1H),7.31(t,J＝7.6Hz,1H),7.24–7.12(m,3H),6.60(dd,J＝5.8,2.3Hz,1H),2.07(s,6H),1.60–1.49(m,1H),1.11–1.02(m,2H),0.91–0.82(m,2H)；MS(ESI)m/z(％):486.2[M+H]⁺,508.1[M+Na]⁺。

EXAMPLE 28N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (3, 4-dimethylphenyl) -1H-imidazole-4-carboxamide (Compound 28)

MS(ESI)m/z(％):486.2[M+H]⁺,508.1[M+Na]⁺。

EXAMPLE 29N- (4- { [2- (acetylamino) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (2-fluorophenyl) -1H-imidazole-4-carboxamide (Compound 29)

¹H NMR(600MHz,DMSO-d₆)δ10.50(s,1H),10.36(s,1H),8.22(d,J＝5.3Hz,2H),8.12(d,J＝5.7Hz,1H),8.01(dd,J＝13.2,2.1Hz,1H),7.83–7.67(m,2H),7.60(s,1H),7.48(dd,J＝8.6,3.6Hz,2H),7.32(d,J＝35.8Hz,2H),6.63(dd,J＝5.7,2.3Hz,1H),1.98(s,3H)；MS(ESI)m/z(％):450.2[M+H]⁺,472.1[M+Na]⁺。

EXAMPLE 30N- (4- { [2- (acetylamino) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-fluorophenyl) -1H-imidazole-4-carboxamide (Compound 30)

¹H NMR(600MHz,DMSO-d₆)δ10.57(s,1H),10.41(s,1H),8.48(d,J＝5.7Hz,2H),8.20(d,J＝5.7Hz,1H),8.09(d,J＝13.2Hz,1H),7.95–7.77(m,3H),7.67(s,1H),7.53–7.27(m,3H),6.71(d,J＝5.4Hz,1H),2.05(s,3H)；MS(ESI)m/z(％):450.1[M+H]⁺,472.1[M+Na]⁺。

EXAMPLE 31N- (4- { [2- (acetylamino) pyridin-4-yl ] oxy } -3-fluorophenyl) -1- (4-methoxyphenyl) -1H-imidazole-4-carboxamide (Compound 31)

IR(KBr,cm^-1):3365.8,3261.6,3130.4,2362.8,1672.3,1516.1,1423.5,1259.5,970.2,823.6,756.1,640.4；¹H NMR(600MHz,DMSO-d₆)δ10.57(s,1H),10.38(s,1H),8.39(s,2H),8.19(d,J＝5.3Hz,1H),8.09(d,J＝13.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.76–7.52(m,3H),7.35(t,J＝8.8Hz,1H),7.10(d,J＝7.9Hz,2H),6.86–6.52(m,1H),3.82(s,3H),2.05(s,3H)；MS(ESI)m/z(％):462.2[M+H]⁺,484.2[M+Na]⁺。

EXAMPLE 32N- (4- { [2- (cyclopropanecarboxamido) pyridin-4-yl ] oxy } phenyl) -1- (4-fluorophenyl) -1H-imidazole-4-carboxamide (Compound 32)

¹H NMR(600MHz,DMSO-d₆)δ10.82(s,1H),10.17(s,1H),8.44(d,J＝5.1Hz,2H),8.18(d,J＝5.7Hz,1H),7.98(d,J＝8.7Hz,2H),7.90–7.75(m,2H),7.65(s,1H),7.42(t,J＝8.6Hz,2H),7.15(d,J＝8.7Hz,2H),6.69(d,J＝5.5Hz,1H),2.12–1.74(m,1H),1.00–0.53(m,4H)；MS(ESI)m/z(％):458.2[M+H]⁺,480.2[M+Na]⁺。

EXAMPLE 33N- (4- { [2- (acetylamino) pyridin-4-yl ] oxy } phenyl) -1- (4-fluorophenyl) -1H-imidazole-4-carboxamide (Compound 33)

¹H NMR(600MHz,DMSO-d₆)δ10.52(s,1H),10.17(s,1H),8.45(d,J＝4.6Hz,2H),8.18(d,J＝5.7Hz,1H),7.98(d,J＝8.9Hz,2H),7.90–7.78(m,2H),7.67(s,1H),7.43(t,J＝8.7Hz,2H),7.16(d,J＝8.9Hz,2H),6.66(dd,J＝5.7,2.3Hz,1H),2.05(s,3H)；MS(ESI)m/z(％):432.2[M+H]⁺,454.1[M+Na]⁺。

Example 34 in vitro anti-tumor cell Activity

Pyridine compounds containing triazole ketoamide and imidazole amide structures are subjected to in-vitro inhibition c-Met kinase activity screening and inhibition human gastric cancer cell strain MKN45, human lung adenocarcinoma cell strain A549, human lung cancer cell H460 and human colon cancer cell HT-29 activity screening.

c-Met enzyme Activity assay

The assay used to measure c-Met kinase activity is based on an enzyme-linked immunosorbent assay (ELISA). The specific operation is as follows:

the example compound, 50pM c-Met (His-tagged recombinant human Met (amino acid 974-terminus), expressed by baculovirus) and 5. mu.M ATP were buffered in the assay at room temperature on 0.25mg/mL PGT coated platesIn solution (25mM MOPS, pH 7.4, 5mM MgCl)₂，0.5raM MnCl₂100 μ M sodium orthovanadate, 0.01% Triton X-100, 1mM DTT, and finally DMSO concentration 1% (v/v)) for 20 minutes. The reaction mixture was removed by washing and the phosphorylated polymer substrate was detected with 0.2. mu.g/mL of a phosphotyrosine-specific monoclonal antibody (PY20) conjugated with horseradish peroxidase (HRP). After the color development was stopped by adding 1M phosphoric acid, the color of the developed substrate (TMB) was quantified spectrophotometrically at 450 nm. The inhibition data of the compounds of the examples on c-Met kinase are shown in table 1.

In vitro cytotoxic Activity assay

(1) After cells were thawed and passaged for 2-3 stabilities, they were digested from the bottom of the flask with trypsin solution (0.25%). After pouring the cell digest into the centrifuge tube, the culture medium is added to stop the digestion. Centrifuging the centrifuge tube at 800r/min for 10min, discarding supernatant, adding 5mL culture solution, blowing and beating the mixed cells, sucking 10 μ L cell suspension, adding into cell counting plate, counting, and adjusting cell concentration to 10⁴Per well. 100. mu.L of the cell suspension was added to the 96-well plate except that the A1 well was a blank well and no cells were added. The 96-well plate was placed in an incubator for 24 h.

(2) The test sample was dissolved in 50. mu.L of dimethyl sulfoxide, and then an appropriate amount of culture solution was added to dissolve the sample to 2mg/mL of the liquid, and then the sample was diluted to 20,4,0.8,0.16, 0.032. mu.g/mL in a 24-well plate.

3 wells were added for each concentration, two columns of cells surrounding each, which were greatly affected by the environment, and only used as blank wells. The 96-well plate was placed in an incubator for 72 h.

(3) The drug-containing culture solution in the 96-well plate is discarded, the cells are washed twice by using Phosphate Buffer Solution (PBS), 100 mu L of MTT (tetrazole) (0.5mg/mL) is added into each well and put into an incubator for 4h, the MTT solution is discarded, and 100 mu L of dimethyl sulfoxide is added. And oscillating on a magnetic oscillator to fully dissolve the viable cells and the MTT reaction product formazan, and putting the formazan into an enzyme labeling instrument to measure the result. Determination of drug IC by Bliss method₅₀The value is obtained. Results of the carried out compound for inhibiting the activity of human lung adenocarcinoma cell line A549, human lung carcinoma cell H460 and human colon carcinoma cell HT-29See table 1.

The results of inhibiting c-Met kinase, human gastric cancer cell strain MKN45, human lung adenocarcinoma cell strain A549, human lung cancer cell H460 and human colon cancer cell HT-29 activity of the compound are shown in Table 1, and IC in Table 1₅₀< 1.0 μ M, expressed as A, 1.0 μ M < IC₅₀< 5.0 μ M, expressed as B, 5.0 μ M < IC₅₀< 10.0 μ M, expressed as C, 10.0 μ M < IC₅₀≦ 100.0. mu.M, denoted by D.

TABLE 1

As is clear from Table 1, the pyridine compounds containing triazole ketoamide and imidazole amide structures and having the general formula (I) to be protected in the invention have good in vitro inhibitory activity on c-Met kinase, human gastric cancer cell strain MKN45, human lung adenocarcinoma cell strain A549, human lung cancer cell H460 and human colon cancer cell HT-29. The compounds have good development and application prospects of antitumor drugs.

While the invention has been described with reference to specific embodiments, modifications and equivalent arrangements will be apparent to those skilled in the art and are intended to be included within the scope of the invention.

Claims

1. Pyridine compounds containing triazole ketoamide and imidazole amide structures shown in general formula (I) and pharmaceutically acceptable salts thereof,

wherein:

x is selected from hydrogen or halogen;

l is selected from

R₁Selected from hydrogen, C₁-C₆Alkyl, or C₃-C₆A cycloalkyl group;

R₂selected from hydrogen, hydroxyl, halogen, nitro, ester group, amino, cyano, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, C halogenated by 1-3 halogens₁-C₆Alkyl, or C halogenated by 1-3 halogens₁-C₆An alkoxy group.

2. The pyridine compound containing triazole ketone amide and imidazole amide structures according to claim 1, and pharmaceutically acceptable salts thereof,

x is selected from hydrogen, fluorine or chlorine;

l is selected from

R₂selected from hydrogen, hydroxyl, halogen, nitro, ester group, amino, cyano, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, tri-propylFluoromethyl, monofluoromethoxy, difluoromethoxy or trifluoromethoxy.

3. The pyridine compound containing triazole ketone amide and imidazole amide structures according to claim 2, and pharmaceutically acceptable salts thereof,

r is selected from methyl, ethyl or cyclopropyl;

x is selected from hydrogen or fluorine;

l is selected from

R₁Selected from hydrogen, methyl, ethyl, or cyclopropyl;

4. The pyridine compound containing triazole ketoamide and imidazole amide structures according to claim 3, and pharmaceutically acceptable salts thereof, characterized by having the following structural formula:

5. a pharmaceutical composition, characterized by comprising the pyridine compounds containing triazole ketone amide and imidazole amide structures and pharmaceutically acceptable salts thereof as active ingredients and pharmaceutically acceptable excipients according to any one of claims 1 to 4.

6. The pyridine compounds containing triazole ketone amide and imidazole amide structures according to any one of claims 1 to 4, pharmaceutically acceptable salts thereof, or the pharmaceutical composition according to claim 5, for use in preparing a medicament for treating and/or preventing proliferative diseases.

7. The pyridine compounds containing triazole ketoamide and imidazole amide structures of any one of claims 1 to 4, pharmaceutically acceptable salts thereof, or the pharmaceutical composition of claim 5 for use in the preparation of c-Met kinase inhibitors.

8. The pyridine compounds containing triazole ketone amide and imidazole amide structures according to any one of claims 1 to 4, pharmaceutically acceptable salts thereof, or the pharmaceutical composition according to claim 5, for use in preparing a medicament for treating and/or preventing cancer.

9. The use according to claim 8, wherein the cancer is gastric cancer, lung cancer and colon cancer.