CN117903128A

CN117903128A - ASK1/PDK1 double-targeting inhibitor, and preparation method and application thereof

Info

Publication number: CN117903128A
Application number: CN202211272362.7A
Authority: CN
Inventors: 李乾斌; 胡高云; 陈卓; 秦佳
Original assignee: Central South University
Current assignee: Central South University
Priority date: 2022-10-18
Filing date: 2022-10-18
Publication date: 2024-04-19

Abstract

The invention belongs to the technical field of medicines, and particularly relates to an ASK1/PDK1 double-targeting inhibitor, pharmaceutically acceptable salts thereof, esters thereof and optical isomers thereof. The compound can be used as an inhibitor of targeted ASK1 and PDK1 and applied to medicines for treating and/or preventing diseases related to ASK1/PDK1 double-targeted mediated fibrosis diseases. The compound is shown in a general formula I, and optical isomer, pharmaceutically acceptable salt and solvate thereof,Wherein R ₁ isR12 is selected from C3-C8 cycloalkyl, methyl, ethyl, C3-C8 straight or branched alkyl; r13 is selected from methyl, ethyl or propyl; r ₂ is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

Description

ASK1/PDK1 double-targeting inhibitor, and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to an ASK1/PDK1 double-targeting inhibitor, pharmaceutically acceptable salts thereof, esters thereof and optical isomers thereof. The compound can be used as an inhibitor of targeted ASK1 and PDK1 and applied to medicines for treating and/or preventing ASK1/PDK1 double-targeted mediated fibrosis diseases.

Background

Apoptosis signal-regulating kinase (ASK 1) is a widely expressed serine/threonine kinase, and knockdown or inhibition of ASK1 is effective in preventing inflammation and fibrosis. Under pathological conditions of cellular stress, ASK1 is activated by stimulation with factors such as Reactive Oxygen Species (ROS), lipopolysaccharide (LPS), tumor necrosis factor-a (TNF-a), and Endoplasmic Reticulum (ER) stress to phosphorylate downstream JNKs and p38 MAPKs proteins, thereby affecting signaling pathways such as inflammation, fibrosis, and apoptosis.

The 3-phosphoinositide dependent protein kinase 1 (PDK 1) is a kinase in AGC protein family, can be highly expressed in endothelial cells, fibroblasts and other cells, and can phosphorylate a downstream factor AKT, thereby realizing the effects of regulating cell proliferation, apoptosis and cell differentiation. Studies show that PDK1 activity is closely related to fibrosis, and inhibition of PDK1 can inhibit cell signal regulation pathways so as to promote apoptosis of fibroblasts.

ASK1 inhibitor GS-4997 was studied clinically on diabetic nephropathy, alcoholic fatty liver and pulmonary arterial hypertension, but clinical trials of GS-4997 single drug treatment did not have significant effects compared to placebo. The individual suppression of one of the signal paths may affect the effect of the other path due to complex interactions between the paths. Therefore, the development direction of ASK1 inhibitors needs to make up for the deficiency of single drug application through single molecule multi-target points. ASK1 and PDK1 have similar regulation effects in cells, but studies have demonstrated that both PDK1 and ASK1 have negative feedback effects. Patel et al report that the combination of a PDK1 inhibitor (BX-795) and an ASK1 inhibitor (GS-4997) can achieve antithrombotic purposes.

The invention patents CN110294746, CN110698471, CN110407806, etc. disclose novel ASK1 inhibitors. So far, the compound only has an inhibiting effect on ASK1, but no related research result exists on ASK1/PDK1 double-targeting inhibitor, so that the ASK1/PDK1 double-targeting inhibitor is an important direction of compound research and has potential research value.

Disclosure of Invention

The invention aims to provide a compound with ASK1/PDK1 double-targeting inhibition effect, which provides possibility for preparing medicines for treating and/or preventing fibrosis diseases mediated by ASK1/PDK1 double-targeting.

The specific technical scheme is as follows:

The invention claims compounds of general formula (i), pharmaceutically acceptable salts, esters and solvates thereof:

Wherein R ₁ is R12 is selected from C3-C8 cycloalkyl, methyl, ethyl, C3-C8 straight or branched alkyl; r13 is selected from methyl, ethyl or propyl;

R ₂ is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

Preferably, R12 is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, isopropyl, n-propyl, n-butyl, isobutyl.

The invention claims compounds of general formula (ii), pharmaceutically acceptable salts, esters and solvates thereof:

wherein R ₃ is R14 is selected from C3-C8 cycloalkyl, methyl, ethyl, C3-C8 straight or branched alkyl;

r ₄ is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

Preferably, R14 is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, isopropyl, n-propyl, n-butyl, isobutyl.

The invention claims compounds of general formula (iii), pharmaceutically acceptable salts, esters and solvates thereof:

wherein X is C or N;

r ₅ is hydrogen, C1-C3 alkyl, halogenated C1-C3 alkyl; r ₆ is hydrogen, C1-C3 alkyl, halogenated C1-C3 alkyl, substituted benzene ring; r ₇ is hydrogen, substituted benzene ring; r ₈ is C3-C8 cycloalkyl, C1-C6 alkyl, C1-C6 hydroxyalkyl.

Preferably, R ₆ is methyl,

Preferably, when R ₇ is a substituted benzene ring, the substituent is one or more of methyl, ethyl, propyl and halogen.

Preferably, when R ₇ is a substituted benzene ring, the substituents are methyl and halogen.

Preferably, when R ₇ is a substituted benzene ring, the methyl and halogen are ortho-substituted on the benzene ring.

Preferably, R ₈ is C3-C6 cycloalkyl, C1-C4 alkyl, C1-C4 hydroxyalkyl.

Preferably, R ₈ is isopropyl, n-propyl, C3 hydroxyalkyl.

Preferably, R ₈ is-CH (CH ₃)CH₂ OH).

The invention claims compounds of general formula (iv), pharmaceutically acceptable salts, esters and solvates thereof:

wherein Y is C or N;

r9 is five-membered heterocycle, substituted five-membered heterocycle, six-membered heterocycle, substituted six-membered heterocycle; the substituent groups on the substituted five-membered heterocycle and the substituted six-membered heterocycle are selected from C3-C8 cycloalkyl, C1-C6 alkyl and C1-C6 haloalkyl;

R10 is hydrogen or substituted benzene ring;

r11 is C3-C8 cycloalkyl, C1-C6 alkyl, C1-C6 hydroxyalkyl.

Preferably, R9 is pyrrole, substituted pyrrole, pyrazole, substituted pyrazole, imidazole, substituted imidazole, pyridine, substituted pyridine, pyrimidine, substituted pyrimidine; the substituents on the substituted pyrrole, substituted pyrazole, substituted imidazole, substituted pyridine and substituted pyrimidine are selected from C3-C8 cycloalkyl, C1-C6 alkyl and C1-C6 haloalkyl.

Preferably, the substituents on the substituted pyrroles, substituted pyrazoles, substituted imidazoles, substituted pyridines and substituted pyrimidines are selected from the group consisting of C3-C6 cycloalkyl, C1-C4 alkyl, C1-C4 haloalkyl.

Preferably, the substituents on the substituted pyrrole, substituted pyrazole, substituted imidazole, substituted pyridine and substituted pyrimidine are selected from isopropyl, n-propyl, n-butyl and isobutyl.

Preferably, R9 is

Preferably, when R ₁₀ is a substituted benzene ring, the substituent is one or more of methyl, ethyl, propyl and halogen.

Preferably, when R ₁₀ is a substituted benzene ring, the substituents are methyl and halogen.

Preferably, when R ₁₀ is a substituted benzene ring, the methyl and halogen are ortho-substituted on the benzene ring.

Preferably, R ₁₁ is C3-C6 cycloalkyl, C1-C4 alkyl, C1-C4 hydroxyalkyl.

Preferably, R ₁₁ is isopropyl, n-propyl, C3 hydroxyalkyl.

Preferably, R ₁₁ is-CH (CH ₃)CH₂ OH).

Preferably, the present invention claims the following compounds, optical isomers, pharmaceutically acceptable salts and solvates thereof, but these compounds are not meant to be limiting in any way to the present invention:

N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide (Compound 01)

N- (6- (4-isopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide (Compound 02)

N- (3- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) phenyl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide (Compound 03)

N- (6- (4-cyclobutyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide (Compound 04)

N- (6- (4-cyclopentyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide (Compound 05)

N- (6- (4-cyclopentyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -7- (2-fluorobenzyl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide (Compound 06)

N- (6- (4-cyclopentyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (4-methoxybenzyl) -6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide (Compound 07)

5- (1-Cyclopropyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxamide (Compound 08)

N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -5- (1-isopropyl-1H-pyrazol-4-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide (Compound 09)

5- (1-Cyclobutyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxamide (Compound 10)

N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -6-oxo-1, 6-dihydro- [3,3' -bipyridine ] -5-carboxamide (Compound 11)

N- (6- (4- (1-hydroxy-prop-2-yl) -4H-1,2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide (Compound 12)

N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -5- (1-isopropyl-1H-pyrazol-4-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide (Compound 13)

5- (1-Cyclopropyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide (Compound 14)

5- (1-Cyclobutyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide (Compound 15)

6- (1-Cyclopropyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) quinoline-4-carboxamide (Compound 16)

5- (1-Cyclobutyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1H-indazole-3-carboxamide (Compound 17)

N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -6- (1-isopropyl-1H-pyrazol-4-yl) quinoline-4-carboxamide (Compound 18)

N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -6- (6-methylpyridin-3-yl) quinoline-3-carboxamide (Compound 19)

N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -5- (1-isopropyl-1H-pyrazol-4-yl) -1H-indazole-3-carboxamide (Compound 20)

5- (1-Cyclobutyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1H-indazole-3-carboxamide (Compound 21)

5- (1-Cyclopropyl-1H-pyrazol-4-yl) -1- (2-fluorobenzyl) -N- (6- (4- (1-hydroxypropyl-2-yl) -4H-1,2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide (Compound 22)

1- (2-Fluorobenzyl) -N- (6- (4- (1-hydroxy-prop-2-yl) -4H-1,2, 4-triazol-3-yl) pyridin-2-yl) -5- (1-isopropyl-1H-pyrazol-4-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide (Compound 23)

The term "pharmaceutically acceptable salts" as used herein refers to pharmaceutically acceptable acid and base addition salts and solvates.

The term "ester" as used herein refers to pharmaceutically acceptable esters, particularly esters which hydrolyze in vivo and include esters which readily decompose in humans to leave the parent compound (compounds of formula (I), (II)) or salts thereof.

The invention also claims processes for the preparation of the compounds of the above general formulae (I-IV), all starting materials being prepared in the manner described in these schemes, by methods known to the person skilled in the art of organic chemistry or being commercially available. All final derivatives of the invention are prepared by the methods described in these schemes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All variables applied in these routes are as defined below or as defined in the claims.

The compounds of the formula (I) can be synthesized in accordance with scheme 1

Step one: intermediate 1 and aromatic heterocyclic carboxylic acid are synthesized through acylation reaction to obtain intermediate 2.

Step two: intermediate 2 reacts with aromatic heterocyclic boric acid ester to obtain the compound shown in the general formula (I).

Wherein R ₁、R₂ is as defined above.

Intermediate 1 was synthesized as in scheme 2:

step one: the compound 1-1 reacts with hydrazine hydrate to generate 1-2

Step two: 1-2 reacts with alkylamine to generate intermediate 1

Wherein R ₂ is isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, halogenated C3-6 cycloalkyl, cyano-substituted C1-4 alkyl or hydroxy-substituted C1-4 alkyl

The compounds of the formula (II) can be synthesized according to scheme 3

The synthesis method of the intermediate 3 and the intermediate 4 is the same as that of the scheme 1.

The compounds of the formula (III) can be synthesized in accordance with scheme 4

The synthesis method of the intermediate 5 in the general formula (III) is the same as that of the route 2, the intermediate 6 is obtained according to the synthetic route 5, firstly, 1-methyl-3- (trifluoromethyl) -1H-pyrazol-5-ol is subjected to Vilsmeier-Haack-Arnold reaction to obtain the compound 6-2, the compound 6-5 is obtained through reduction and cyclization reaction, and finally, the intermediate 6 is obtained through substitution and decarboxylation.

Wherein R ₅ is selected from CH ₃、CF₃;R₆ is selected from CH ₃,R ₇ is selected from hydrogen, CH ₃、CH₃CH₂,

The compounds of the formula (IV) can be synthesized according to scheme 6

The synthesis method of the intermediate 7 and the intermediate 8 is the same as that of the scheme 1.R ₉、R₁₀、R₁₁ is as defined above.

The invention also claims the application of the compounds represented by the general formulas (I-IV) in medicines for treating and/or preventing ASK1/PDK1 double-targeting mediated related diseases.

Preferably, the ASK1/PDK1 dual-targeting mediated related disease is fibrosis.

The invention also provides a pharmaceutical composition comprising the aforementioned compound, a pharmaceutically acceptable salt, ester or stereoisomer thereof.

In some embodiments of the invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers and/or diluents.

The present invention also provides a pharmaceutical formulation comprising a compound as described above, a pharmaceutically acceptable salt, ester or stereoisomer thereof, and one or more pharmaceutically acceptable carriers and/or diluents; the pharmaceutical preparation is any clinically or pharmaceutically acceptable dosage form.

In some embodiments of the invention, the above-described pharmaceutical formulations may be administered orally, parenterally, rectally, or pulmonary, etc., to a patient or subject in need of such treatment. For oral administration, the pharmaceutical composition may be formulated into oral preparations, for example, into conventional oral solid preparations such as tablets, capsules, pills, granules, etc.; can also be made into oral liquid preparation such as oral solution, oral suspension, syrup, etc. When the composition is formulated into oral preparations, suitable fillers, binders, disintegrants, lubricants, etc. may be added. For parenteral administration, the pharmaceutical preparations may also be formulated as injections, including injectable solutions, injectable sterile powders and injectable concentrated solutions. When the injection is prepared, the conventional method in the existing pharmaceutical field can be adopted for production, and when the injection is prepared, no additive can be added, and the proper additive can be added according to the property of the medicine. For rectal administration, the pharmaceutical composition may be formulated as suppositories and the like. For pulmonary administration, the pharmaceutical composition may be formulated as an inhalant or spray, etc.

The pharmaceutically acceptable carrier and/or diluent useful in the pharmaceutical composition or pharmaceutical formulation of the present invention may be any conventional carrier and/or diluent in the pharmaceutical formulation arts, and the choice of the particular carrier and/or diluent will depend on the mode of administration or type and state of disease for the particular patient being treated. The preparation of suitable pharmaceutical compositions for specific modes of administration is well within the knowledge of those skilled in the pharmaceutical arts. For example, pharmaceutically acceptable carriers and/or diluents may include solvents, diluents, dispersing agents, suspending agents, surfactants, isotonic agents, thickening agents, emulsifying agents, binders, lubricants, stabilizers, hydration agents, emulsifying accelerators, buffers, absorbents, colorants, ion-exchange agents, mold release agents, coating agents, flavoring agents, antioxidants and the like which are conventional in the pharmaceutical arts. Flavoring agent, antiseptic, sweetener, etc. can be added into the pharmaceutical composition if necessary.

The invention has the advantages that:

1. the compound synthesized by the invention has better inhibitory activity on ASK1/PDK 1.

2. The compound synthesized by the invention can achieve the effect of inhibiting the excessive proliferation of the fibrotic cells by inhibiting ASK1/PDK1 double channels, and can be applied to the aspect of related medicaments.

Drawings

FIG. 1 shows the effect of the compounds of the present invention on PDK1 pathway.

FIG. 2 shows the effect of the compounds of the invention on AKT pathway.

Detailed Description

The technical solution of the present invention will be described below with reference to specific embodiments, and the described embodiments are only some embodiments of the present invention, but not all embodiments.

Representative compounds of the invention are shown in table 1:

TABLE 1 representative Compounds of the invention

Example 1

Synthesis of Compound 1

1.1 Preparation of 6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-amine

Step one: compound 6-aminopicolinate (20 g,132 mmol) is dissolved in 250ml methanol, hydrazine hydrate solution (13 g,260 mmol) is added for reaction for 8h at 80 ℃, after the reaction is finished, the reaction solution is cooled to 0 ℃, suction filtration is carried out, the obtained solid is washed by methanol, and the target product is obtained by drying, wherein the yield is 80%.

Step two: the compound 6-aminopyridine formylhydrazine (5 g,26 mmol) and triethyl orthoformate were dissolved in 100ml of 1, 4-dioxane and reacted at 75 ℃ for 2 hours, cyclopropylamine (1.87 g,33 mmol), acetic acid (1.97 g,33 mmol) and 100 ℃ were added sequentially, and after the starting materials were reacted completely, the reaction solution was concentrated, and the residue was chromatographed on silica gel (CH ₃ OH/dcm=1:30) to give the target compound 5.16g in 78% yield.

Preparation of N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide

1-Methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxylic acid (500 mg,1.92 mmol) was dissolved in 10ml thionyl chloride, reacted at 40℃for 2 hours, the solvent concentrated, 3- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2 amine (150 mg,0.74 mmol) was added, 15ml dichloromethane was dissolved, and reacted at room temperature for 5 hours. After the reaction was completed, the reaction solution was concentrated, and the residue was subjected to column chromatography (ch3oh/dcm=1:30) to give 50mg of the objective compound in the yield of 19％.¹H NMR(400MHz,DMSO-d₆)δ14.01(s,1H),8.68(s,1H),8.58(s,1H),8.36(d,J＝8.2Hz,1H),8.00(t,J＝8.0Hz,1H),7.83(d,J＝7.6Hz,1H),3.89(s,3H),3.87–3.81(m,1H),1.22–1.15(m,2H),1.10–1.03(m,2H).¹³C NMR(151MHz,DMSO-d₆)δ＝164.15,152.62,151.86,146.20,145.96,139.87,133.64,122.82,121.04,118.39,115.71,114.30,102.32,35.01,29.41,8.20.ESI m/z:445.1348[M+H]⁺;Found:445.1348

Example 2

Synthesis of Compound 2

Preparation of N- (6- (4-isopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide

The intermediate 6- (4-isopropyl-4H-1, 2, 4-triazole-3-yl) pyridin-2-amine was prepared in 80% yield by the method of step 1.1 in example 1, starting from intermediate 6-aminopyridine formylhydrazine and isopropylamine by cyclization.

The procedure of step 1.2 of example 1 was followed using as starting material intermediate 6- (4-isopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-amine and 1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxylic acid to give the title compound in the following yield 21％.¹H NMR(400MHz,DMSO-d₆)δ14.00(s,1H),12.62(s,1H),8.92(s,1H),8.72(s,1H),8.34(d,J＝8.3Hz,1H),8.05(s,1H),7.86(d,J＝7.6Hz,1H),5.44–5.32(m,1H),4.00(s,3H),1.55(d,J＝6.7Hz,6H).¹³C NMR(151MHz,DMSO-d₆)δ＝163.47,158.77,150.07,143.51,142.15,139.85,112.53,111.06,49.36,34.61,34.52,23.37.ESI m/z:447.1505[M+H]⁺;Found:447.1516

Example 3

Synthesis of Compound 3

Preparation of N- (3- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) phenyl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazole [3,4-b ] pyridine-5-carboxamide

3.1 Preparation of 6- (4-isopropyl-4H-1, 2, 4-triazol-3-yl) aniline

Step one: dissolving compound 6-amino-2 methyl benzoate (30 g, 199mmol) in 200ml methanol, adding hydrazine hydrate solution (21 g,420 mmol), reacting at 80 ℃ for 8h, cooling the reaction solution to 0 ℃ after the reaction is finished, filtering, washing the obtained solid with methanol, drying to obtain target product 25g, yield 83％.¹H NMR(400MHz,DMSO-d6)δ9.53(s,1H),7.13–6.98(m,2H),6.92(d,J＝7.7Hz,1H),6.67(dd,J＝7.9,1.6Hz,1H),5.23(s,2H),4.41(s,2H).

Step two: the compound 3-aminobenzoyl hydrazine (4 g,26 mmol) and triethyl orthoformate were dissolved in 100ml of 1, 4-dioxane and reacted at 75℃for 2 hours, isopropylamine (1.24 g,21 mmol), acetic acid (5 g,83 mmol) were added sequentially, reacted at 100℃for 12 hours, and after the starting material was reacted completely, the reaction solution was concentrated, and the residue was subjected to silica gel column chromatography (methanol: dichloromethane=1:30) to give the objective compound 3.79g in 82% yield.

The procedure of step 1.2 of example 1 was followed using 3- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) aniline as a starting material and 1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxylic acid to give the title compound in the following yield 21％.¹H NMR(400MHz,DMSO-d₆)δ12.82(s,1H),8.62(d,J＝8.0Hz,2H),8.37(d,J＝15.1Hz,1H),7.81(d,J＝8.0Hz,1H),7.68(d,J＝7.8Hz,1H),7.54(t,J＝7.9Hz,1H),3.96(s,3H),3.65(ddd,J＝10.9,7.3,3.9Hz,1H),1.12–1.03(m,2H),0.99–0.92(m,2H).¹³C NMR(126MHz,DMSO-d₆)δ＝163.93,159.73,152.79,151.43,149.48,146.14,145.63,142.47,140.13,135.91,131.32,130.89,124.77,123.78,119.35,115.84,114.43,112.71,110.55,62.94,33.71,28.64,12.56,7.63.ESI m/z:444.1396[M+H]⁺;Found:444.1416

Example 4

Synthesis of Compound 4

Preparation of N- (6- (4-cyclobutyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide

The intermediate 3- (4-cyclobutyl-4H-1, 2, 4-triazol-3-yl) aniline was prepared by cyclization with cyclobutylamine using the intermediate 6-aminopyridine formylhydrazine as the starting material according to the procedure of step 1.1 in example 1, with a yield of 80%. .

The procedure of step 1.2 of example 1 was followed using 3- (4-cyclobutyl-4H-1, 2, 4-triazol-3-yl) aniline as a starting material and 1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxylic acid to give the title compound in the following yield 16％.¹H NMR(400MHz,DMSO-d₆)δ13.88(s,1H),12.81(s,1H),8.98(s,1H),8.67(s,1H),8.31(d,J＝8.2Hz,1H),8.00(t,J＝8.0Hz,1H),7.83(d,J＝7.6Hz,1H),5.41–5.24(m,1H),3.97(s,3H),3.17(s,2H),2.63(dd,J＝16.4,7.9Hz,2H),1.94–1.72(m,2H).¹³C NMR(101MHz,DMSO-d₆)δ＝162.37,151.28,150.71,146.30,144.63,140.30,136.02,118.94,116.28,114.20,102.73,50.92,49.01,36.20,31.11,14.88.ESI m/z:459.1505[M+H]⁺;Found:459.1525

Example 5

Synthesis of Compound 5

Preparation of N- (6- (4-cyclopentyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide

The intermediate 6- (4-cyclopentyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-amine was prepared in 76% yield by the procedure of step 1.1 of example 1 starting from intermediate 6-aminopyridine formylhydrazine and cyclopentylamine.

The procedure of step 1.2 of example 1 was followed using as starting material intermediate 6- (4-cyclopentyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-amine and 1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxylic acid to give the title compound in yield 37％.¹H NMR(400MHz,DMSO-d₆)δ13.29(s,1H),8.88(s,1H),8.63(s,1H),8.35(d,J＝8.2Hz,1H),8.01(t,J＝8.0Hz,1H),7.84(d,J＝7.6Hz,1H),5.43(m,J＝14.4,7.2Hz,1H),3.95(s,3H),2.35–2.22(m,2H),1.95–1.82(m,4H),1.74(s,2H).¹³C NMR(151MHz,DMSO-d₆)δ＝170.38,169.34,159.62,152.31,151.96,146.61,146.23,143.98,143.34,140.24,138.41,131.21,114.33,113.57,111.95,108.91,57.97,34.27,33.71,23.84.ESI m/z:473.1661[M+H]⁺;Found:473.1684

Example 6

Synthesis of Compound 6

Preparation of N- (6- (4-cyclopentyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -7- (2-fluorobenzyl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide

The procedure of step 1.2 of example 1 was followed using as starting material intermediate 6- (4-cyclopentyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-amine and 7- (2-fluorobenzyl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxylic acid to give the title compound in the yield of 65％.¹H NMR(400MHz,DMSO-d₆)δ10.81(s,1H),8.82(s,1H),8.61(s,1H),8.27(d,J＝8.3Hz,1H),8.04(s,1H),7.86(d,J＝7.5Hz,1H),7.69(ddd,J＝23.2,7.6,6.1Hz,1H),7.38–7.30(m,1H),7.23–7.16(m,1H),7.07(s,1H),5.71(s,2H),5.57–5.47(m,1H),4.14(s,3H),2.07(dd,J＝13.5,5.4Hz,2H),1.81–1.67(m,4H),1.59(dd,J＝10.1,5.9Hz,2H).¹³C NMR(126MHz,DMSO-d₆)δ＝164.47,163.75,161.66,161.34,160.47,159.70,151.37,151.00,149.44,148.98,146.68,143.94,140.24,135.04,133.19,130.86,130.78,130.75,130.72,125.01,124.99,124.81,123.79,123.68,122.61,115.84,115.67,114.65,113.12,106.48,62.95,62.92,57.28,52.89,34.93,34.88,33.69,23.97.ESI m/z:581.2037[M+H]⁺;Found:581.2059

Example 7

Synthesis of Compound 7

Preparation of N- (6- (4-cyclopentyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (4-methoxybenzyl) -6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide

The procedure of step 1.2 of example 1 was followed using as starting material intermediate 6- (4-cyclopentyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-amine and 1- (4-methoxybenzyl) -6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxylic acid to give the title compound in the yield of 44％.¹H NMR(400MHz,DMSO)δ14.09(s,1H),12.58(s,1H),8.88(s,1H),8.72(s,1H),8.36(d,J＝8.3Hz,1H),8.05(t,J＝8.0Hz,1H),7.88(d,J＝7.6Hz,1H),7.32(d,J＝8.6Hz,2H),6.94(d,J＝8.6Hz,2H),5.55(s,2H),5.48–5.36(m,1H),3.73(s,3H),2.36–2.23(m,2H),1.91(dd,J＝12.5,5.7Hz,4H),1.74(s,2H).¹³C NMR(126MHz,DMSO)δ＝159.62,151.15,150.95,146.70,144.01,140.45,129.90,127.74,119.56,116.87,114.62,114.35,102.88,57.99,55.60,51.59,33.73,23.89.

Example 8

Synthesis of Compound 8

8.1 Preparation of 5-bromo-1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid

5-Bromo-2-oxo-1, 2-dihydropyridine-3-carboxylic acid (10 g,31 mmol), 1- (bromomethyl) -2-fluorobenzene (17 g,90 mmol) and potassium carbonate (12 g,87 mmol) were dissolved in acetonitrile (300 ml) and reacted at 70℃for 6h. After the reaction is finished, concentrating the solution, precipitating solid, and carrying out suction filtration to obtain 12g of target product with yield 79％.¹H NMR(500MHz,DMSO-d₆)δ13.97(s,1H),8.72(d,J＝2.3Hz,1H),8.38(d,J＝2.4Hz,1H),7.39(dd,J＝13.7,6.9Hz,1H),7.32–7.11(m,3H),5.33(s,2H).

8.2 Preparation of 5-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxamide

5-Bromo-1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid (2 g,6 mmol), 6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-amine (2.5 g,12 mmol) and HATU (3.5 g,9 mmol) were added to DMF (15 ml), reacted at room temperature, and DIPEA (1.6 g,12 mmol) was added to react for 4H at 40 ℃. At the end of the reaction, 30ml of water were added, the solid precipitated, suction filtration was carried out to obtain a crude product, which was dissolved with CH ₃ OH and purified by silica gel column chromatography (DCM/CH ₃ oh=30:1) to obtain 2.3g of the target compound in 74% yield .¹H NMR(400MHz,DMSO-d₆)δ12.40(s,1H),8.71–8.66(m,2H),8.51(d,J＝2.8Hz,1H),8.29(d,J＝8.2Hz,1H),8.02(t,J＝8.0Hz,1H),7.86(d,J＝7.6Hz,1H),7.38(s,1H),7.31(s,1H),7.25(d,J＝10.0Hz,1H),7.20(d,J＝7.6Hz,1H),5.32(s,2H),3.76(s,1H),1.14–1.06(m,2H),1.03(dd,J＝5.8,3.3Hz,2H).

8.3 Preparation of 5- (1-cyclopropyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxamide

5-Bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxamide (200 mg,0.39 mmol), 1-cyclopropyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (200 mg,0.7 mmol), bis triphenylphosphine palladium dichloride (88 mg,0.1 mmol) and cesium carbonate (128 mg,4 mmol) were dissolved in 1, 4-dioxane and water and heated to 100deg.C under nitrogen for reaction for 4H. At the end of the reaction, the solution was concentrated, extracted with DCM (30 ml x 3), dried over anhydrous sodium sulfate, filtered, and dried by spin-drying to give the crude compound 129mg,61% yield by silica gel column chromatography (DCM/ch3oh=30:1) .¹H NMR(400MHz,DMSO-d₆)δ12.66(s,1H),8.75(d,J＝2.7Hz,1H),8.69–8.65(m,2H),8.37–8.29(m,2H),8.03(t,J＝8.0Hz,1H),7.85(d,J＝6.8Hz,2H),7.38(td,J＝7.4,1.6Hz,1H),7.26(dd,J＝6.7,4.8Hz,2H),7.19(d,J＝7.7Hz,1H),5.37(s,2H),3.77(dd,J＝7.4,3.6Hz,2H),1.16–1.06(m,4H),1.05–0.95(m,4H).¹³C NMR(101MHz,DMSO-d₆)δ＝162.08,160.94,152.55,151.21,146.30,146.08,142.72,140.40,140.16,135.96,130.28,130.25,127.25,125.06,125.02,123.42,123.28,119.85,119.20,116.94,115.98,115.77,114.34,113.38,48.25,48.21,33.34,29.19,8.09,6.76.ESI m/z:537.2163[M+H]⁺;Found:537.2184

Example 9

Synthesis of Compound 9

Preparation of N- (6- (4-isopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -5- (1-isopropyl-1H-pyrazol-4-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide

The procedure of step 8.3 of example 8 was followed using 5-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxamide as starting material to make suzuki reaction with 1-isopropyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole as starting material to give the title compound in the yield of 33％.¹H NMR(400MHz,DMSO-d₆)δ12.68(s,1H),8.76(d,J＝2.7Hz,1H),8.67(d,J＝3.9Hz,2H),8.37–8.30(m,2H),8.04(t,J＝8.0Hz,1H),7.88–7.83(m,2H),7.38(td,J＝7.4,1.7Hz,1H),7.27(d,J＝9.2Hz,2H),7.19(d,J＝7.5Hz,1H),5.38(s,2H),4.52(s,1H),3.77(s,1H),1.46(d,J＝6.7Hz,6H),1.14–1.10(m,2H),1.06–1.01(m,2H).¹3C NMR(101MHz,DMSO)δ＝162.13,161.87,160.94,159.43,152.57,151.23,146.32,146.06,142.74,140.27,140.16,135.40,130.47,130.38,130.26,130.22,125.05,123.46,123.32,119.85,119.22,116.74,115.98,115.77,114.35,113.69,53.74,48.20,29.49,29.17,23.13,8.08.ESI m/z:539.2319[M+H]⁺;Found:539.2331.

Example 10

Synthesis of Compound 10

Preparation of 5- (1-cyclobutyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxamide

The procedure of step 8.3 of example 8 was followed using 5-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxamide as starting material to make suzuki reaction with 1-cyclobutyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole to give the title compound in the yield of 61％.¹H NMR(400MHz,DMSO-d₆)δ12.66(s,1H),8.76(d,J＝2.6Hz,1H),8.68(s,2H),8.40–8.31(m,2H),8.03(t,J＝8.0Hz,1H),7.93–7.82(m,2H),7.43–7.34(m,1H),7.30–7.14(m,3H),5.37(s,2H),4.98–4.61(m,1H),3.89–3.63(m,1H),2.50–2.36(m,4H),1.82(td,J＝10.0,4.3Hz,2H),1.19–0.92(m,4H).¹³C NMR(151MHz,DMSO-d₆)δ＝162.12,160.93,152.55,151.21,146.30,146.06,142.73,140.27,140.16,135.39,130.44,130.39,130.23,130.21,125.04,123.43,119.83,119.21,116.73,115.94,115.80,114.34,113.68,53.73,48.22,29.18,23.14,8.08.ESI m/z:551.2319[M+H]⁺;Found:551.2345.

Example 11

Synthesis of Compound 11

Preparation of N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -6-oxo-1, 6-dihydro- [3,3' -bipyridine ] -5-carboxamide

The title compound was prepared by following the procedure of step 8.3 in example 8 by clazuki reaction of intermediate 5-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1- (2-fluorobenzyl) -2-oxo-1, 2-dihydropyridine-3-carboxamide starting from 3- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridine in the yield of 16％.¹H NMR(500MHz,DMSO-d₆)δ12.69(s,1H),11.24(s,1H),8.86(d,J＝2.8Hz,1H),8.73–8.65(m,2H),8.36(d,J＝8.3Hz,1H),8.03(t,J＝8.0Hz,1H),7.89–7.81(m,2H),7.52(d,J＝8.4Hz,1H),7.42(t,J＝2.7Hz,1H),7.40–7.31(m,3H),7.22(dt,J＝15.0,8.6Hz,2H),6.52(s,1H),5.46(s,2H),3.84–3.74(m,1H),1.18–1.09(m,2H),1.07–1.00(m,2H).¹³C NMR(151MHz,DMSO-d₆)δ＝162.25,161.08,152.57,151.24,146.30,146.06,143.63,141.80,140.15,135.96,130.66,130.63,130.50,130.44,128.77,126.98,126.26,125.03,123.44,121.80,119.72,119.45,119.20,117.86,115.96,115.82,114.38,112.64,101.95,29.20,8.10.ESI m/z:546.2054[M+H]⁺;Found:546.2071.

Example 12

Synthesis of Compound 12

Preparation of N- (6- (4- (1-hydroxy-prop-2-yl) -4H-1,2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide

12.1 Preparation of 2- (3- (6-aminopyridin-2-yl) -4H-1,2, 4-triazol-4-yl) propan-1-ol

Step one: compound 6-aminopicolinate (50 g,329 mmol) is dissolved in 200ml methanol, hydrazine hydrate solution (32 g,420 mmol) is added for reaction for 8h at 80 ℃, after the reaction is finished, the reaction solution is cooled to 0 ℃, suction filtration is carried out, the obtained solid is washed by methanol, and the target product is obtained by drying, wherein the yield is 84%.

Step two: the compound 6-aminopyridine formylhydrazine (5 g,26 mmol) and triethyl orthoformate are dissolved in 100ml of 1, 4-dioxane to react for 2 hours at 75 ℃, 2-aminopropan-1-ol (2.5 g,33 mmol) and acetic acid (5 g,83 mmol) are added in sequence, the reaction is carried out for 12 hours at 100 ℃, after the raw materials are reacted completely, the reaction solution is concentrated, and the residue is chromatographed on a silica gel column (methanol: dichloromethane=1:30) to obtain the target compound 5.62g, the yield is 78％.¹H NMR(400MHz,DMSO-d₆)δ8.69(s,1H),7.52(dd,J＝8.3,7.3Hz,1H),7.18(dd,J＝7.4,0.9Hz,1H),6.52(dd,J＝8.3,0.9Hz,1H),6.17(s,1H),5.50–5.28(m,1H),5.03(s,1H),3.72–3.51(m,1H),1.43(d,J＝6.9Hz,2H).

Preparation of N- (6- (4- (1-hydroxypropan-2-yl) -4H-1,2, 4-triazol-3-yl) pyridin-2-yl) -1-methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxamide

1-Methyl-6-oxo-3- (trifluoromethyl) -6, 7-dihydro-1H-pyrazolo [3,4-b ] pyridine-5-carboxylic acid (500 mg,1.92 mmol) was dissolved in 10ml thionyl chloride, reacted at 40℃for 2 hours, the solvent concentrated, 2- (3- (6-aminopyridin-2-yl) -4H-1,2, 4-triazol-4-yl) propan-1-ol (420 mg,2 mmol) was added, dichloromethane (15 ml) was dissolved, and reacted at room temperature for 5 hours. After the reaction was completed, the reaction solution was concentrated, and the residue was subjected to column chromatography (CH ₃ OH/dcm=1:30) to give 100mg of the objective compound in yield 11％.¹H NMR(400MHz,DMSO)δ15.25(s,1H),8.79(s,1H),8.46(s,1H),8.36(d,J＝8.0Hz,1H),7.96(t,J＝8.0Hz,1H),7.74(d,J＝7.5Hz,1H),5.33(d,J＝5.2Hz,1H),5.12(s,1H),3.80(s,3H),3.73(s,1H),1.89(s,2H),1.54(d,J＝6.9Hz,3H).¹³C NMR(101MHz,DMSO)δ＝170.57,165.83,154.30,152.42,151.15,146.65,144.21,139.88,131.30,118.41,114.96,114.20,64.66,54.51,33.71,29.28,18.08.

Example 13

Synthesis of Compound 13

Preparation of 5- (1-cyclopropyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide

The title compound was prepared by following the procedure of step 8.3 in example 8 using 5-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide as starting material and by suzuki reaction with 1-cyclopropyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole in the yield of 57％.¹H NMR(600MHz,DMSO-d₆)δ13.01(d,J＝6.4Hz,1H),12.90(s,1H),8.69(d,J＝2.7Hz,2H),8.36(d,J＝8.2Hz,1H),8.30(s,1H),8.11(dd,J＝6.4,2.8Hz,1H),8.04(t,J＝8.0Hz,1H),7.90–7.85(m,2H),3.82(tt,J＝7.6,4.1Hz,1H),3.73(tt,J＝7.4,3.8Hz,1H),1.17(td,J＝7.4,5.5Hz,2H),1.11–1.04(m,4H),0.98(td,J＝7.4,4.9Hz,2H).¹³C NMR(151MHz,DMSO)δ＝162.40,162.05,152.49,151.33,146.28,146.11,142.99,140.15,136.25,136.08,127.06,119.78,118.94,117.11,114.29,113.45,40.40,40.26,40.12,39.98,39.84,39.70,39.57,33.30,29.40,8.20,6.76.

Example 14

Synthesis of Compound 14

Preparation of N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -5- (1-isopropyl-1H-pyrazol-4-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide

The title compound was prepared by following the procedure of step 8.3 in example 8 using 5-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide as starting material and by suzuki reaction with 1-isopropyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole in the yield of 54％.¹H NMR(600MHz,DMSO-d₆)δ13.00(d,J＝6.3Hz,1H),12.91(s,1H),8.74–8.64(m,2H),8.36(d,J＝8.3Hz,1H),8.30(s,1H),8.12(dd,J＝6.4,2.8Hz,1H),8.04(t,J＝8.0Hz,1H),7.88(t,J＝3.8Hz,2H),4.49(hept,J＝6.7Hz,1H),3.82(tt,J＝7.6,4.1Hz,1H),1.45(d,J＝6.7Hz,5H),1.17(td,J＝7.4,5.5Hz,2H),1.09–1.03(m,2H).¹³C NMR(151MHz,DMSO)δ＝162.43,162.04,152.51,151.34,146.31,146.11,142.99,140.15,136.12,135.48,124.90,119.76,118.94,116.92,114.29,113.76,53.66,40.40,40.26,40.12,39.98,39.84,39.71,39.57,30.62,29.89,29.38,23.15,8.19.

Example 15

Synthesis of Compound 15

Preparation of 5- (1-cyclobutyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide

The title compound was prepared by following the procedure of step 8.3 in example 8 using 5-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide as starting material and performing suzuki reaction with 1-cyclobutyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole in the yield of 16％.¹H NMR(600MHz,DMSO-d₆)δ13.04–12.99(m,1H),12.91(d,J＝5.6Hz,1H),8.73–8.64(m,2H),8.36(d,J＝8.0Hz,2H),8.16–8.11(m,1H),8.04(t,J＝8.0Hz,1H),7.91(s,1H),7.88(d,J＝7.6Hz,1H),4.83(p,J＝8.4Hz,1H),3.82(tt,J＝7.6,4.1Hz,1H),2.47(td,J＝9.5,2.7Hz,1H),2.40(qt,J＝7.7,2.9Hz,2H),1.80(ddt,J＝14.7,13.0,5.5Hz,2H),1.17(td,J＝7.4,5.5Hz,2H),1.11–1.02(m,2H).

Example 16

Synthesis of Compound 16

Preparation of 6- (1-cyclopropyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) quinoline-3-carboxamide

The procedure of step 8.3 of example 8 was followed using intermediate 6-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) quinoline-3-carboxamide as starting material was followed by suzuki reaction with 1-cyclopropyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole to give the title compound in the yield of 26％.¹H NMR(600MHz,DMSO-d₆)δ11.12(s,1H),9.26(d,J＝2.2Hz,1H),8.85(d,J＝2.2Hz,1H),8.67(s,1H),8.47(s,1H),8.34–8.31(m,2H),8.17(dd,J＝8.8,2.0Hz,1H),8.11–8.05(m,3H),7.86(dd,J＝7.6,0.9Hz,1H),4.30(tt,J＝7.5,4.1Hz,1H),3.80(tt,J＝7.5,3.8Hz,1H),1.14–1.10(m,2H),1.06–1.01(m,4H),0.97–0.93(m,2H).¹³C NMR(151MHz,DMSO)δ＝165.83,152.90,151.85,148.72,147.96,146.14,145.41,139.95,137.09,136.59,132.18,130.31,129.74,128.35,128.13,127.39,123.72,121.14,119.54,115.34,33.40,28.81,7.65,6.82.ESI m/z:463.1995[M+H]⁺;463.1990.

Example 17

Synthesis of Compound 17

Preparation of 6- (1-cyclobutyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) quinoline-3-carboxamide

The procedure of step 8.3 of example 8 was followed using intermediate 6-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) quinoline-3-carboxamide as starting material was followed by suzuki reaction with 1-cyclobutyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole to give the title compound in the yield of 37％.¹H NMR(600MHz,DMSO-d₆)δ11.12(s,1H),9.26(d,J＝2.3Hz,1H),8.86(d,J＝2.2Hz,1H),8.67(s,1H),8.51(d,J＝0.8Hz,1H),8.33(dd,J＝8.3,1.1Hz,2H),8.18(dd,J＝8.8,2.0Hz,1H),8.13–8.05(m,3H),7.86(dd,J＝7.6,0.8Hz,1H),4.94–4.83(m,1H),4.30(tt,J＝7.6,4.1Hz,1H),2.54(dd,J＝9.8,2.6Hz,1H),2.47–2.40(m,2H),1.91–1.80(m,2H),1.04(td,J＝7.4,5.4Hz,2H),0.97–0.91(m,2H).¹³C NMR(151MHz,DMSO)δ＝165.84,152.90,151.85,148.71,147.96,146.15,145.41,139.96,137.07,136.60,132.32,130.33,129.75,128.14,127.41,127.00,123.71,121.22,119.55,115.34,55.44,40.40,40.26,40.12,39.98,39.84,39.70,39.57,30.59,28.81,14.61,7.65.ESI m/z:477.2151[M+H]⁺;477.2148.

Example 18

Synthesis of Compound 18

Preparation of N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -6- (1-isopropyl-1H-pyrazol-4-yl) quinoline-3-carboxamide

The procedure of step 8.3 of example 8 was followed using intermediate 6-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) quinoline-3-carboxamide as starting material and 1-isopropyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole to give the title compound in the form of a suzuki reaction 22％.¹H NMR(600MHz,DMSO-d₆)δ11.11(s,1H),9.26(d,J＝2.2Hz,1H),8.86(d,J＝2.2Hz,1H),8.67(s,1H),8.46(s,1H),8.33(dd,J＝5.2,3.2Hz,2H),8.18(dd,J＝8.7,2.0Hz,1H),8.12–8.04(m,3H),7.86(dd,J＝7.6,0.8Hz,1H),4.56(hept,J＝6.7Hz,1H),4.30(tt,J＝7.5,4.1Hz,1H),1.49(d,J＝6.6Hz,5H),1.04(td,J＝7.4,5.2Hz,2H),0.96(ddd,J＝7.3,5.6,4.3Hz,2H).¹³C NMR(151MHz,DMSO)δ＝165.85,152.90,151.85,148.64,147.92,146.15,145.41,139.95,136.59,136.58,132.49,130.32,129.73,128.13,127.43,126.12,123.57,120.91,119.54,115.34,53.77,40.40,40.26,40.12,39.98,39.85,39.71,39.57,28.81,23.17,7.65.ESI m/z:465.2151[M+H]⁺;465.2145.

Example 19

Synthesis of Compound 19

Preparation of N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -6- (6-methylpyridin-3-yl) quinoline-3-carboxamide

The procedure of step 8.3 of example 8 was followed using as starting material intermediate 6-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) quinoline-3-carboxamide, followed by clazuki reaction with 2-methyl-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridine to give the title compound in yield 24％.¹H NMR(400MHz,DMSO)δ11.18(s,1H),9.37(d,J＝2.2Hz,1H),9.03(d,J＝2.0Hz,1H),8.97(d,J＝2.2Hz,1H),8.68(s,1H),8.54(d,J＝1.9Hz,1H),8.35–8.32(m,1H),8.30–8.26(m,1H),8.22(d,J＝8.8Hz,1H),8.18(dd,J＝8.1,2.5Hz,1H),8.13–8.05(m,1H),7.87(dd,J＝7.6,0.7Hz,1H),7.45(d,J＝8.1Hz,1H),4.31(s,1H),2.56(s,3H),1.07–1.01(m,2H),1.00–0.92(m,2H).

Example 20

Synthesis of Compound 20

Preparation of N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -5- (1-isopropyl-1H-pyrazol-4-yl) -1H-indazole-3-carboxamide

The procedure of step 8.3 of example 8 was followed using 5-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1H-indazole-3-carboxamide as a starting material and by suzuki reaction with 1-isopropyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole to give the title compound in the yield of 13％.¹H NMR(500MHz,DMSO-d₆)δ13.26(s,1H),10.86(s,1H),8.68(d,J＝6.7Hz,2H),8.48(s,2H),8.32(d,J＝8.3Hz,1H),8.10(s,1H),8.04(t,J＝8.0Hz,1H),7.98(dd,J＝8.7,1.6Hz,1H),7.81(d,J＝7.6Hz,1H),7.64(d,J＝8.8Hz,1H),4.62(p,J＝6.7Hz,1H),4.32(tt,J＝7.4,4.1Hz,1H),1.50(d,J＝6.6Hz,6H),1.06–0.99(m,2H),0.97(dq,J＝7.5,4.5,3.9Hz,2H).ESI m/z:454.2104[M+H]⁺:454.2133.

Example 21

Synthesis of Compound 21

Preparation of 5- (1-cyclobutyl-1H-pyrazol-4-yl) -N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1H-indazole-3-carboxamide

The procedure of step 8.3 of example 8 was followed using 5-bromo-N- (6- (4-cyclopropyl-4H-1, 2, 4-triazol-3-yl) pyridin-2-yl) -1H-indazole-3-carboxamide as a starting material and performing suzuki reaction with 1-cyclobutyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole to give the title compound in the yield of 32％.¹H NMR(600MHz,DMSO-d₆)δ13.27(s,1H),10.86(s,1H),8.72–8.65(m,2H),8.55(s,1H),8.32(dd,J＝8.4,0.9Hz,1H),8.13(s,1H),8.05(t,J＝8.0Hz,1H),7.98(dd,J＝8.7,1.5Hz,1H),7.82(dd,J＝7.6,0.9Hz,1H),7.65(d,J＝8.7Hz,1H),5.00–4.89(m,1H),4.31(tt,J＝7.5,4.1Hz,1H),2.69–2.51(m,4H),2.47–2.37(m,2H),1.90–1.76(m,2H),1.11–0.91(m,4H).ESI m/z:466.2104[M+H]⁺:466.2124.

Example 22

Synthesis of Compound 22

Preparation of 5- (1-cyclopropyl-1H-pyrazol-4-yl) -1- (2-fluorobenzyl) -N- (6- (4- (1-hydroxypropyl-2-yl) -4H-1,2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide

The procedure of step 8.3 of example 8 was followed using 5-bromo-1- (2-fluorobenzyl) -N- (6- (4- (1-hydroxypropyl-2-yl) -4H-1,2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide as starting material and bell-mouthing with 1-cyclopropyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole to give the title compound in the yield of 36％.¹H NMR(400MHz,DMSO-d₆)δ12.66(s,1H),8.33(d,J＝8.7Hz,1H),8.04(t,J＝8.0Hz,1H),7.38(t,J＝7.0Hz,1H),7.34–7.22(m,1H),5.37(s,1H),5.20(q,J＝6.1Hz,1H),5.02(s,1H),3.76(tt,J＝7.3,3.8Hz,1H),3.72(d,J＝5.5Hz,1H),1.49(d,J＝6.9Hz,1H),1.08(m,J＝3.3,2.9Hz,1H),1.05–0.94(m,1H).

Example 23

Synthesis of Compound 23

Preparation of 1- (2-fluorobenzyl) -N- (6- (4- (1-hydroxypropan-2-yl) -4H-1,2, 4-triazol-3-yl) pyridin-2-yl) -5- (1-isopropyl-1H-pyrazol-4-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide

The procedure of step 8.3 of example 8 was followed using 5-bromo-1- (2-fluorobenzyl) -N- (6- (4- (1-hydroxypropyl-2-yl) -4H-1,2, 4-triazol-3-yl) pyridin-2-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide as starting material and bell-mouthing with 1-isopropyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole to give the title compound in the yield of 45％.¹H NMR(400MHz,DMSO-d₆)δ12.68(s,1H),8.80(s,1H),8.75(d,J＝2.7Hz,1H),8.68(d,J＝2.7Hz,1H),8.37–8.28(m,2H),8.04(t,J＝8.0Hz,1H),7.87(d,J＝0.8Hz,1H),7.81(dd,J＝7.7,0.9Hz,1H),7.45–7.33(m,1H),7.34–7.10(m,3H),5.38(s,2H),5.20(h,J＝6.6Hz,1H),5.02(t,J＝5.4Hz,1H),4.52(h,J＝6.6Hz,1H),3.77–3.66(m,2H),1.47(dd,J＝12.8,6.8Hz,9H).

Biological examples

The beneficial effects of the compounds of the invention are further illustrated below by biological experiments, but this should not be understood as the compounds of the invention only having the following beneficial effects.

Example 24:

In vitro enzymatic Activity of the Compounds of the invention

Test article: the chemical structural formula of the compound synthesized by the embodiment of the invention is shown in table 1, and the experimental materials are shown in table 2.

Table 2 experimental materials

The experimental method comprises the following steps:

1. Dilution of the Compounds

1) The compounds of the invention were diluted to 8mM using DMSO.

2) The above-described compound of the present invention was diluted to 500. Mu.M or 1mM. The compounds of the invention were diluted in a two-fold gradient to 12 concentrations, the highest concentration being 5 μm or 10 μm.

3) Mu.L of the compound of the present invention was transferred to 384 well plates using a pipette, respectively.

2. Preparation of a reaction system

1) An enzyme reaction system comprising a kinase and factors required for the reaction was prepared according to Table 3

TABLE 3 enzymatic reaction System

2) 2UL of the reaction enzyme system was added to each reaction well

3) To each reaction well was added 2ul of 2.5 x ATP/Substrate Mix system (see table 4);

TABLE 4 2.5 XATP/Substrate Mix system

4) After the reaction system is placed at 37 ℃ for reaction for 60min, 5 mu L of ADP-Glo ^TM Reagent is added and incubated at 37 ℃ for 40min;

5) Adding 10 mu L Kinase Detection Reagent, incubating at 37 ℃ for 30min, and detecting by a multifunctional enzyme-labeled instrument.

3. Data analysis:

3.1 is calculated using the following formula:

% enzyme remaining = (average value of OD value of test group/OD value of blank group) ×100%

Enzyme inhibition = (1-average OD value of test group/OD value of blank group) ×100%

And (3) carrying out regression-Probit method calculation by using SPSS statistical analysis software to obtain the IC ₅₀ value.

3.2 Results are as follows:

compounds of table 5 for ASK1 inhibitory activity

Sequence number	ASK1 IC₅₀(μM)	Sequence number	ASK1 IC₅₀(μM)
				Compound 1	0.49	Compound 14	0.004
Compound 2	2.66	Compound 15	0.002
				Compound 3	2.70	Compound 16	0.519
Compound 4	0.98	Compound 17	0.13
				Compound 5	2.18	Compound 18	0.068
Compound 6	1.02	Compound 19	1.22
				Compound 7	0.708	Compound 20	0.161
Compound 8	0.002	Compound 21	0.50
				Compound 9	0.009	Compound 22	0.007
Compound 10	0.28	Compound 23	0.011
				Compound 11	2.37	GS-444217	0.007
Compound 12	0.179	MP7	>5
				Compound 13	0.005

Compounds of table 6 have PDK1 inhibitory activity

Sequence number	PDK1 inhibition@10μΜ(％)	Sequence number	PDK1 inhibition@10μΜ(％)
				Compound 1	27.45	Compound 14	22.11
Compound 2	42.85	Compound 15	19.78
				Compound 3	27.96	Compound 16	26.03
Compound 4	31.94	Compound 17	18.54
				Compound 5	37.72	Compound 18	27.72
Compound 6	29.90	Compound 19	24.35
				Compound 7	14.20	Compound 20	46.11
Compound 8	13.63	Compound 21	39.19
				Compound 9	23.80	Compound 22	12.99
Compound 10	40.27	Compound 23	17.49
				Compound 11	20.99	GS-444217	0.04
Compound 12	44.38	MP7	65.09
				Compound 13	32.19

The test results show that: the compound prepared by the invention can effectively inhibit ASK1 and PDK1 activities, and has obvious double-targeting inhibition. The positive drug GS-444217 has inhibition effect on ASK1 only and has no inhibition effect on PDK 1. MP7 was inhibitory to PDK1 alone and had no inhibitory effect on ASK 1.

Example 25

In vitro cytological inhibitory Activity of the Compounds of the invention

Test compound:

1) Test article: preferred compounds of the invention have the chemical structures shown in Table 1

2) Positive drug: GS-444217, MP7

Experimental materials:

1) Cell lines: human embryonic lung fibroblasts (HFL-1), rat kidney fibroblasts (NRK-49F), rat liver-shaped cells (HSC-T6)

2) Reagents and instrumentation: 96 well plates (Corning); fetal bovine serum (Gibco), F12K medium, DMEM medium (BI); a biosafety cabinet, a carbon dioxide incubator (ESCO); multifunctional enzyme-labeled instrument (Bio-tek).

The experimental method comprises the following steps:

1) Cell culture: the experiments were performed with cells in the logarithmic growth phase and with a proportion of viable cells higher than 90%.

2) Cell growth detection: cells were digested, diluted to a cell suspension at a concentration of 3-5×10 ⁴ cells/mL, 100 μl of cell suspension (3-5×10 ³ cells per well) was added to 60 wells in the middle of the 96-well plate, and 100 μl of PBS was added to 36 wells around; culturing 96-well plate in incubator with 5% CO2 at 37deg.C until it adheres to wall (enters logarithmic phase); after sucking up the medium in the middle 60 wells, 100 μl of medium containing different concentrations of compound was added to each well, 3 wells per group; placing the 96-well plate in an incubator for culturing for 48 hours; the medium containing the compound was discarded, and PBS was added to 100. Mu.L of a 10% CCK-8 base medium per well of the 96-well plate, and after incubating the 96-well plate in the incubator for 1-4 hours, the OD was measured at 450nm using an ELISA reader.

Data analysis:

the proliferation rate and inhibition rate of each group of cells were calculated.

Cell viability% = (average value of OD value of test group/OD value of blank group) ×100%

Inhibition = (1-average OD value of test group/OD value of blank group) ×100%

Calculating half effective inhibition concentration (IC ₅₀) of the drug, and calculating IC ₅₀ by regression-inhibition method using SPSS statistical analysis software according to drug action concentration and growth inhibition rate of the drug on cell action.

The experimental results are shown in table 7:

TABLE 7 growth inhibition of cell action by the compounds of the invention

Compared with a control drug, the compound prepared by the invention has better inhibitory activity on three fibroblasts of lung, kidney and liver and is obviously better than a positive drug, so that the compound shown in the general formula (I) or (II) provided by the invention is an effective ASK1/PDK1 inhibitor.

Example 26

Effect of the inventive Compounds on ASK1/PDK1 pathway

Test compound:

3) Test article: preferred compounds 8 and 9 of the present invention have the chemical structures shown in Table 1

Experimental materials:

1) Cell lines: human embryonic lung fibroblast (HFL-1)

2) Reagents and instrumentation: 6-well plate (Corning); fetal bovine serum (Gibco), F12K medium; a biosafety cabinet, a carbon dioxide incubator (ESCO); multifunctional enzyme labelling instrument (Bio-tek); chemiluminescent imaging system (Bio-Rad); and a steady voltage and steady flow electrophoresis apparatus (Bio-Rad).

The experimental method comprises the following steps:

1) 8% of separating gel and laminating gel are configured according to the gel concentration and volume, electrophoresis liquid (preferably completely penetrating through the laminating gel) is added, the sample is loaded, the loading amount of the sample is 30 mug/lane, and the Marker is 5 mug. And (3) laminating the gel electrophoresis voltage of 80V, adjusting the voltage to 120V to continue electrophoresis when the sample uniformly runs to the separation gel, and stopping until bromophenol blue is about to run out of the gel.

2) The membrane transferring filter paper is prepared, the foam cushion, the two layers of filter paper, the gel, the PVDF membrane, the two layers of filter paper and the foam cushion are placed in sequence, the gel is arranged at the negative electrode, the PVDF membrane is arranged at the positive electrode, and constant current transfer is carried out for 2 hours.

3) Washing the membrane with TBST for 3 times, each time for 5-10 min, transferring into TBST containing 5% skimmed milk powder for sealing, shaking at room temperature for 1-2 h, and sucking the sealing liquid with filter paper.

4) Adding primary antibody: the murine anti-human actin monoclonal antibody was diluted 1:5000 overnight at 4 ℃ with an anti-dilution.

5) Adding a secondary antibody: goat anti-rabbit HRP-lgG was diluted 1:5000 with TBST and shaken for 1h at room temperature.

6) Preparing chemiluminescent liquid according to the ratio of 1:1: each of the reagent A and the reagent B was taken in 0.1mL. And (3) fully contacting the membrane protein surface downwards with the mixed luminous liquid, and exposing for 1-3 min.

The experimental results are shown in fig. 1 and 2, and PDK1 can phosphorylate its downstream factor AKT, thereby realizing the effect of regulating cell proliferation, apoptosis and cell differentiation. Experimental results show that after HFL-1 cells are stimulated by TGF-beta 1, the phosphorylation level of PDK1 and AKT proteins is increased, and after compound 8 and compound 9 act for 24 hours, the phosphorylation of PDK1 and the phosphorylation level of AKT can be obviously inhibited, and the effect of PDK1/AKT channels can be effectively regulated. This experiment further verifies that compound 8 has a positive PDK1 inhibitory effect at a 10. Mu.M inhibition of 13%.

As shown in FIG. 2, the results of detecting the ASK1 pathway by the compound 8 and the compound 9 show that the phosphorylation levels of ASK1, P38 and JNK proteins are increased after HFL-1 cells are stimulated by TGF-beta 1, and the phosphorylation levels of ASK1, P38 and JNK can be obviously inhibited after the compound 8 and the compound 9 act for 24 hours, so that the effect of the ASK1/P38/JNK pathway can be effectively regulated. The Western Blot experiment proves that the positive medicine GS-444217 has no inhibition effect on PDK1 and AKT phosphorylation, MP7 has no obvious inhibition effect on ASK1, P38 and JNK, and the compound 8 and the compound 9 can inhibit ASK1/PDK1 double channels at the same time, and the result is consistent with the enzyme experiment result.

Claims

1. A compound shown as a general formula I and an optical isomer, pharmaceutically acceptable salt and solvate thereof,

Wherein R ₁ isR12 is selected from C3-C8 cycloalkyl, methyl, ethyl, C3-C8 straight or branched alkyl; r13 is selected from methyl, ethyl or propyl;

R ₂ is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

2. The compound according to claim 1, wherein R12 is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, isopropyl, n-propyl, n-butyl, isobutyl, and optical isomers, pharmaceutically acceptable salts, solvates thereof.

3. A compound of formula (ii), pharmaceutically acceptable salts, esters and solvates thereof:

r ₄ is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

4. The compound according to claim 2, wherein R14 is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, isopropyl, n-propyl, n-butyl, isobutyl, and optical isomers, pharmaceutically acceptable salts, solvates thereof.

5. A compound of formula (iii), pharmaceutically acceptable salts, esters and solvates thereof:

wherein X is C or N;

6. The compound according to claim 5, wherein R ₆ is methyl, and optical isomers, pharmaceutically acceptable salts, solvates thereof,

7. The compound according to claim 5, wherein when R ₇ is a substituted benzene ring, the substituent is one or more of methyl, ethyl, propyl, and halogen; preferably, when R ₇ is a substituted benzene ring, the substituents are methyl and halogen.

8. The compound of claim 5, wherein R ₈ is C3-C6 cycloalkyl, C1-C4 alkyl, C1-C4 hydroxyalkyl, and optical isomers, pharmaceutically acceptable salts, solvates thereof; preferably, R ₈ is isopropyl, n-propyl, C3 hydroxyalkyl.

9. A compound of formula (iv), pharmaceutically acceptable salts, esters and solvates thereof:

wherein Y is C or N;

R10 is hydrogen or substituted benzene ring;

r11 is C3-C8 cycloalkyl, C1-C6 alkyl, C1-C6 hydroxyalkyl.

10. The compound of claim 9, wherein R9 is pyrrole, substituted pyrrole, pyrazole, substituted pyrazole, imidazole, substituted imidazole, pyridine, substituted pyridine, pyrimidine, substituted pyrimidine, and optical isomers, pharmaceutically acceptable salts, solvates thereof; the substituents on the substituted pyrrole, substituted pyrazole, substituted imidazole, substituted pyridine and substituted pyrimidine are selected from C3-C8 cycloalkyl, C1-C6 alkyl and C1-C6 haloalkyl.

11. The compound of claim 10, wherein the substituents on the substituted pyrrole, substituted pyrazole, substituted imidazole, substituted pyridine and substituted pyrimidine are selected from the group consisting of C3-C6 cycloalkyl, C1-C4 alkyl, C1-C4 haloalkyl; preferably, the substituents on the substituted pyrrole, substituted pyrazole, substituted imidazole, substituted pyridine and substituted pyrimidine are selected from isopropyl, n-propyl, n-butyl and isobutyl.

12. The compound according to any one of claims 9-11, wherein R9 is

13. The compound according to claim 9, wherein, when R ₁₀ is substituted benzene ring, the substituent is one or more of methyl, ethyl, propyl, halogen; when R ₁₀ is a substituted benzene ring, the substituent is methyl and halogen.

14. The compound of claim 9, wherein R ₁₁ is C3-C6 cycloalkyl, C1-C4 alkyl, C1-C4 hydroxyalkyl, and optical isomers, pharmaceutically acceptable salts, solvates thereof; preferably, R ₁₁ is isopropyl, n-propyl, C3 hydroxyalkyl.

15. A compound according to claims 1-14, an optical isomer, a pharmaceutically acceptable salt, a solvate thereof, said compound being selected from the group consisting of:

1- (2-Fluorobenzyl) -N- (6- (4- (1-hydroxypropan-2-yl) -4H-1,2, 4-triazol-3-yl) pyridin-2-yl) -5- (1-isopropyl-1H-pyrazol-4-yl) -2-oxo-1, 2-dihydropyridine-3-carboxamide (compound 23).

16. A method of synthesizing a compound according to claim 1 or 2, comprising the steps of:

Step one: the intermediate 1 and aromatic heterocyclic carboxylic acid are synthesized through an acylation reaction to obtain an intermediate 2;

step two: intermediate 2 reacts with aromatic heterocyclic boric acid ester to obtain a compound shown in a general formula (I);

The synthetic route is as follows:

17. Use of a compound according to any one of claims 1-15, a pharmaceutically acceptable salt, ester or stereoisomer thereof for the manufacture of a medicament for the treatment and/or prophylaxis of ASK1/PDK1 dual-target related diseases.

18. The use according to claim 17, wherein the ASK1/PDK1 dual target disease is fibrosis.