ALK5 inhibitor as well as preparation method and application thereof

Abstract

The invention discloses an ALK5 inhibitor as well as a preparation method and application thereof. The ALK5 inhibitor is a compound as shown in a formula I. The compound as shown in the formula I is a novel, efficient and organ-restricted ALK5 inhibitor, has excellent ALK5 inhibitory activity, has an inhibitory IC50 value of 0.879-5.23 nmol / L for ALK5 activity, has the characteristic of low systemic exposure risk, and can be used for preparing an ALK5 inhibitor. The compound has a wide potential of being developed to treat gastrointestinal fibrosis diseases, gastrointestinal tumors, pulmonary fibrosis diseases and fibrotic skin and eye diseases.

Description

Technical Field

[0001] This invention belongs to the field of medicine, and specifically relates to an ALK5 inhibitor, its preparation method, and its application. Background Technology

[0002] Transforming growth factor-β (TGF-β) is a superfamily of cytokines that includes TGFβs, activins, inhibins, Nodal signaling, bone morphogenetic proteins (BMPs), anti-Müllerian hormone (AMH), and growth and differentiation factors (GDFs). These cytokines regulate a wide range of physiological functions, including cell proliferation, differentiation, attachment, migration, and apoptosis.

[0003] TGF-β has a strong fibrosis-inducing effect; it is an activator of fibroblasts, which are the main source of type I collagen and other fibrotic factors. In tumor cells, TGF-β can directly affect tumor growth, or indirectly affect tumor growth by inducing epithelial-mesenchymal transition, blocking anti-tumor immune responses, increasing tumor-associated fibrosis, and enhancing angiogenesis.

[0004] Signal transduction in TGF-β superfamily members requires two related transmembrane heteropolymer serine / threonine kinase receptors, referred to as Type I receptor (TGFβRI) or ALK5 (Activin receptor-like kinase 5) and Type II receptor, respectively. Activation of TGFβRI / ALK5 mediates at least two independent signal continuation and transduction pathways: the SMAD-dependent canonical pathway and the SMAD-independent or non-canonical pathway. Alterations in any part of the TGF-β / Smads signaling pathway will lead to abnormalities in the signal transduction pathway.

[0005] TGFβRI / ALK5 is a key node in TGFβ signaling, and the development of potent, low-toxicity, and selective ALK5 inhibitors is currently a hot topic in the pharmaceutical industry, especially for the treatment of tumors and fibrotic diseases. Currently, Eli Lilly's ALK5 inhibitor LY2157299 (Galunisertib) (WO2002094833) failed in a phase III clinical trial for the treatment of glioma and liver cancer. Medpacto's ALK5 inhibitor EW-7197 is in a phase II clinical trial. LY2157299 and EW-7197 can inhibit tumor cell invasion and metastasis, while also inhibiting tumor cell infiltration into blood vessels. However, existing ALK5 inhibitors inevitably interfere with normal physiological functions, and systemic exposure will produce serious toxicities, including (1) cardiovascular toxicity, which is the most serious and common dose-limiting toxicity, such as aortic aneurysm and dissection, which are life-threatening emergencies; (2) hematologic toxicity, such as bleeding tendency, anemia, and neutropenia; and (3) liver toxicity, such as elevated transaminase levels. The systemic exposure-related toxicities of ALK5 inhibitors severely limit their clinical development and application. Summary of the Invention

[0006] In order to overcome at least one technical problem existing in the prior art, one object of the present invention is to provide a compound represented by Formula I; its general structural formula is as follows:

[0007] .

[0008] A second objective of this invention is to provide a method for preparing the compound represented by Formula I above.

[0009] A third objective of this invention is to provide a pharmaceutical composition.

[0010] A fourth objective of this invention is to provide the use of the compound represented by Formula I or the pharmaceutical composition thereof in the preparation of a medicament for the prevention or treatment of diseases that are improved by inhibiting transforming growth factor-β receptor I / ALK5.

[0011] To achieve the above objectives, the technical solution adopted by the present invention is as follows: The first aspect of the present invention is to provide a compound represented by Formula I,

[0012] Or its pharmaceutically acceptable salts, esters, deuterated compounds, isotopically labeled derivatives, solvates, crystalline compounds, stereoisomers, and prodrugs; Wherein, ring A is selected from 5-10 member heteroaryl groups; the heteroaryl group is optionally unsubstituted or substituted with one or more R groups. a replace; Ring B is selected from 8- to 15-membered heteroaryl groups, wherein ring B contains at least one heteroatom selected from N, S, and O; the heteroaryl group is optionally unsubstituted or substituted with one or more R groups. a replace; When the heteroatom in ring B is N and the number of heteroatoms is 1, Y is not CH; Each R1 is independently selected from -COOR c Halogen, cyano, carboxyl, sulfonium pentafluoride, C 1-8 Alkyl, C 1-8 Alkoxy or C 3-8 Cycloalkyl; the alkyl, alkoxy, or cycloalkyl group is optionally unsubstituted or substituted with one or more R groups. b replace; R2 is selected from hydrogen, halogen, or C. 1-8 Alkyl; the alkyl group is optionally unsubstituted or substituted with one or more R b replace; Each R a Each is independently selected from halogens, deuterium, and carbon. 1-8 Alkyl, C 3-8 cycloalkyl or C 1-8 Alkoxy; the alkyl, alkoxy, or cycloalkyl group is optionally unsubstituted or substituted with one or more R groups. b replace; R c C 1-8 Alkyl; the alkyl group is optionally unsubstituted or substituted with one or more R b replace; Each R b Each is independently selected from halogens or deuterium; X is selected from O or S; Y is selected from CH or N; n is an integer selected from 0 to 4.

[0013] In some embodiments of the present invention, each R1 is independently selected from ester, halogen, cyano, carboxyl, sulfonium pentafluoride, C 1-6 Alkyl, C 1-6 Alkoxy or C 3-6 Cycloalkyl; in some embodiments of the invention, each R1 is independently selected from H, fluorine, chlorine, bromine, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoroethyl, cyano, carboxyl, methyl, deuterated methyl, ethyl, propyl, isopropyl, cyclopropyl, tert-butyl, monofluoromethoxy, difluoromethoxy, trifluoromethoxy, methoxy, deuterated methoxy, ethoxy, propoxy, or isopropoxy.

[0014] In some embodiments of the present invention, R2 is selected from hydrogen, fluorine, chlorine, bromine, or halogen-substituted C. 1-5 alkyl.

[0015] In some embodiments of the present invention, each R a Each is independently selected from fluorine, chlorine, bromine, methyl, deuterated methyl, ethyl, propyl, isopropyl, cyclopropyl, monofluoromethoxy, difluoromethoxy, trifluoromethoxy, methoxy, deuterated methoxy, ethoxy, propoxy, or isopropoxy.

[0016] In some embodiments of the present invention, ring A is selected from five- to eight-membered heteroaryl groups; in some embodiments of the present invention, ring A is selected from five- to six-membered heteroaryl groups; in some embodiments of the present invention, ring A is selected from a thiazole ring or a pyridine ring; in some embodiments of the present invention, ring A is selected from... or R3 is selected from H, fluorine, chlorine, bromine, methyl, deuterated methyl, ethyl or propyl.

[0017] In some embodiments of the present invention, ring B is selected from 6-membered 6-membered or 6-membered 5-membered heteroaryl groups; in some embodiments of the present invention, ring B is selected from benzothiazole, naphthidine, 2,1,3-benzothiadiazole, [1,2,4]triazolo[1,5-A]pyridine, quinoxaline, benzopyridine, thiophene[3,2-C]pyridine, or 1,4-benzodioxane; in some embodiments of the present invention, ring B is selected from... , , , , , , or .

[0018] In some embodiments of the present invention, the compound represented by Formula I is selected from:

[0019]

[0020]

[0021]

[0022]

[0023]

[0024]

[0025]

[0026] .

[0027] The second aspect of the present invention provides a method for preparing the compound represented by Formula I as described in the first aspect of the present invention. When X in the compound represented by Formula I is O, the preparation method includes the following steps: The compound of formula Ia is reacted with the compound of formula Ib to obtain the compound shown in formula I; And optionally, it also includes the following steps: The compound of formula I obtained is reacted with Lawson's reagent to give the compound of formula I with X as S; The structural formula of the compound of formula Ia is: ; The structural formula of the compound of formula Ib is: ; Wherein, Z is selected from Br or Cl; Rings A, B, R1, R2, X, Y, and n are as defined above.

[0028] In some embodiments of the present invention, the compound of formula Ib is obtained by making... It is prepared by reacting with bromoacetyl bromide or chloroacetyl chloride.

[0029] A third aspect of the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound of formula I as described in the first aspect of the invention, or a pharmaceutically acceptable salt thereof, its ester, its deuterated compound, its isotopically labeled derivative, its solvate, its crystalline compound, its stereoisomer, or its prodrug.

[0030] In some embodiments of the present invention, the dosage form of the pharmaceutical composition is selected from tablets, lozenges, tablets, aqueous suspensions, oil suspensions, water-oil suspensions, powders, granules, emulsions, hard capsules, soft capsules, syrups, or elixirs.

[0031] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one of the following conventional pharmaceutical solvents, inert diluents, dispersants, granulating agents, surfactants, emulsifiers, disintegrants, binders, preservatives, buffers, lubricants, excipients, colorants, coating agents, sweeteners, flavoring agents, and aromatizers.

[0032] In some embodiments of the present invention, the pharmaceutical compositions of the present invention are suitable for multiple routes of administration and can thus be formulated into any pharmaceutically acceptable dosage form. For example, the above-described pharmaceutical compositions can be administered to patients or subjects requiring such treatment via oral, parenteral, rectal, or pulmonary administration. When used for oral administration, the pharmaceutical compositions can be formulated into oral preparations, such as conventional oral solid preparations, like tablets, capsules, pills, granules, etc.; or into oral liquid preparations, such as oral solutions, oral suspensions, syrups, etc. When formulating oral preparations, suitable fillers, binders, disintegrants, lubricants, etc., can be added. When used for parenteral administration, the above-described pharmaceutical compositions can also be formulated into injections, including injection solutions, sterile powders for injection, and concentrated solutions for injection. When formulating injections, conventional methods in the existing pharmaceutical field can be used; when formulating injections, excipients may not be added, or suitable excipients may be added depending on the properties of the drug. When used for rectal administration, the pharmaceutical compositions can be formulated into suppositories, etc. When used for pulmonary administration, the pharmaceutical compositions can be formulated into inhaled preparations, aerosols, powder inhalers, or sprays, etc.

[0033] In some embodiments of the invention, pharmaceutically acceptable excipients are substances that are non-toxic, compatible with the active ingredient, and otherwise biologically suitable for use in organisms. The selection of a specific excipient will depend on the route of administration or the type and state of disease for treating a particular patient. Examples of pharmaceutically acceptable excipients include, but are not limited to, solvents, inert diluents, dispersants and / or granulators, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants, and / or oils conventional in the pharmaceutical field. The composition may also contain excipients such as cocoa butter and suppository waxes, colorants, coating agents, sweeteners, flavoring agents, and aromatizers.

[0034] Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and mixtures thereof.

[0035] Exemplary granulating agents and / or dispersants include potato starch, corn starch, cassava starch, sodium starch glycolate, clay, alginate, guar gum, citrus pomace, agar, bentonite, cellulose and wood products, natural sponges, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinylpyrrolidone) (cross-linked polyvinylpyrrolidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (cross-linked carboxymethyl cellulose), methyl cellulose, pregelatinized starch (starch 1500), microcrystalline starch, water-insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof.

[0036] Exemplary surfactants and / or emulsifiers include natural emulsifiers (e.g., gum arabic, agar, alginic acid, sodium alginate, tragacanth gum, carrageenan, cholesterol, xanthan gum, pectin, gelatin, egg yolk, casein, lanolin, cholesterol, waxes, and lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminum silicate)), long-chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, glyceryl triacetate monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxylated polymethylene, polyacrylic acid, acrylic polymers, and carboxyethylene polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethyl cellulose, powdered cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate (20), poly... ethylene-oxygenated sorbitol (60), polyoxyethylene-oxygenated sorbitol monooleate (80), sorbitol monopalmitate (40), sorbitol monostearate (60), sorbitol tristearate (65), glyceryl monooleate, sorbitol monooleate (80), polyoxyethylene esters (e.g., polyoxyethylene monostearate (45), polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate), sucrose fatty acid esters, polyethylene glycol fatty acids Esters (e.g.), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether (30)), poly(ethylene-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, F-68, poloxamer P-188, cetrimonium bromide, cetylpyridine chloride, benzalkonium chloride, docusatesodium and / or mixtures thereof.

[0037] Exemplary binders include starch (e.g., corn starch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., gum arabic, sodium alginate, Irish moss extract, panwar gum, ghatti gum, mucilage of isapola husk, carboxymethyl cellulose, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, microcrystalline cellulose, cellulose acetate, poly(vinylpyrrolidone), magnesium aluminum silicate, and larch arabinogalactan), alginate, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethyl methacrylate, waxes, water, ethanol, and / or mixtures thereof.

[0038] Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, antiprotozoal preservatives, alcohol preservatives, acid preservatives, and other preservatives. In some embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent.

[0039] Exemplary antioxidants include alpha-tocopherol, ascorbic acid, ascorbate palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.

[0040] Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and its salts and hydrates (e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, etc.), citric acid and its salts and hydrates (e.g., citric acid monohydrate), fumaric acid and its salts and hydrates, malic acid and its salts and hydrates, phosphoric acid and its salts and hydrates, and tartaric acid and its salts and hydrates.

[0041] Exemplary antimicrobial preservatives include benzalkonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridine chloride, chlorhexidine, chlorobutanol, chlorocresol, xylene, cresol, ethanol, glycerin, hexetidine, imidureurium, phenol, phenoxyethanol, phenethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.

[0042] Exemplary buffers include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium gluconate, calcium gluconate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propionic acid, calcium fructate, valeric acid, calcium hydrogen phosphate, phosphoric acid, trivalent calcium phosphate, calcium hydroxide, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginate, pyrogen-free raw water, isotonic saline, Ringer's solution, ethanol, and mixtures thereof.

[0043] Exemplary lubricants include magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behenate, hydrogenated vegetable oil, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.

[0044] Exemplary natural oils include bitter almond oil, almond oil, avocado oil, babassu coconut oil, bergamot oil, blackcurrant seed oil, borage oil, juniper oil, chamomile oil, canola oil, caraway oil, Brazilian wax palm oil, castor oil, cinnamon oil, cocoa butter, coconut oil, cod liver oil, coffee oil, corn oil, cottonseed oil, emu oil, eucalyptus oil, evening primrose oil, fish oil, flaxseed oil, geraniol oil, gourd oil, grapeseed oil, hazelnut oil, hyssop oil, isopropyl myristate, jojoba oil, kukui argan oil, and mixed lavender oil. Lavender oil, lemon oil, litsea cubeba oil, macadamia nut oil, mallow oil, mango seed oil, meadowfoam seed oil, mink oil, nutmeg oil, olive oil, orange oil, orange sea bream oil, palm oil, palm kernel oil, peach kernel oil, peanut oil, poppy seed oil, pumpkin seed oil, rapeseed oil, rice bran oil, rosemary oil, safflower oil, sandalwood oil, camellia oil, peppermint oil, sea buckthorn oil, sesame oil, shea butter, silicone oil, soybean oil, sunflower oil, tea tree oil, thistle oil, ailanthus oil, vetiver oil, walnut oil, and wheat germ oil.

[0045] Exemplary synthetic oils include, but are not limited to, butyl stearate, octanoic acid triglyceride, caprylic acid triglyceride, cyclomethicone, diethyl sebacate, polydimethylsiloxane 360, isopropyl myristate, mineral oil, octyl dodecanol, oleyl alcohol, silicone oil, and mixtures thereof.

[0046] Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, liquid dosage forms may also contain inert diluents (such as water or other solvents), solubilizers, and emulsifiers commonly used in the art, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, methyl benzoate, propylene glycol, 1,3-butanediol, dimethylformamide, oils (e.g., cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitol, and mixtures thereof. In addition to inert diluents, oral compositions may include adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and aromatizers.

[0047] In some embodiments for parenteral administration, the compositions described herein are mixed with solubilizers such as alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof. Injectable formulations (e.g., sterile injectable aqueous or oily suspensions) can be formulated using suitable dispersants or wetting agents and suspending agents according to known techniques. Sterile injectable formulations can be sterile injectable solutions, suspensions, or emulsions in non-toxic, parenteral-acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable media and solvents that can be used are water, Ringer's solution (USP), and isotonic sodium chloride solutions. Furthermore, sterile, fixed oils are conventionally used as solvents or suspension media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. Additionally, fatty acids such as oleic acid are used in the preparation of injectables. Injectable formulations may be sterilized, for example, by filtration through a bacterial trap or by incorporating a sterilizing agent in the form of a sterile solid composition dissolved or dispersed in sterile water or other sterile injectable media prior to use.

[0048] To prolong the action of a drug, it is often necessary to slow down the absorption of drugs injected subcutaneously or intramuscularly. This can be achieved by using liquid suspensions of crystalline or amorphous materials with poor water solubility. The absorption rate of the drug depends on its dissolution rate, which in turn depends on the crystal size and crystal form. Alternatively, delayed absorption of parenteral drug forms can be achieved by dissolving or suspending the drug in an oil-based medium. Compositions for rectal or vaginal administration are typically suppositories, which can be prepared by mixing the composition described herein with a suitable non-irritating excipient or carrier (such as cocoa butter, polyethylene glycol, or suppository wax) that is solid at ambient temperature but liquid at body temperature, and thus melts in the rectal or vaginal cavity and releases the active ingredient.

[0049] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharmaceutically acceptable excipient or carrier (such as sodium citrate or dicalcium phosphate) and / or the following substances: (a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato or cassava starch, alginate, certain silicates, and sodium carbonate; (e) dissolution inhibitors, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) absorbents, such as kaolin and bentonite; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may include a buffer. Similar types of solid compositions can be used as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose or lactose and high molecular weight polyethylene glycol. Solid dosage forms of tablets, sugar pills, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the field of pharmacology. They may optionally contain opaque agents and may have compositions that release only one or more active ingredients or preferentially, optionally in a delayed manner, one or more active ingredients in a portion of the intestine. Examples of encapsulation compositions that can be used include polymeric substances and waxes. Similar types of solid compositions can be used as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose or lactose and high molecular weight polyethylene glycol.

[0050] The active ingredient may be microencapsulated together with one or more excipients as mentioned above. Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules may be prepared with coatings and shells, such as enteric coatings, controlled-release coatings, and other coatings well known in the field of pharmaceutical formulation. In such solid dosage forms, the active ingredient may be mixed with at least one inert diluent (such as sucrose, lactose, or starch). Such dosage forms may (as is generally the case) contain substances other than inert diluents, such as tablet lubricants and other tablet excipients, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pellets, the dosage form may contain a buffer. They may optionally contain an opaque agent and may have a composition that releases only one or more active ingredients or preferentially, optionally in a delayed manner, one or more active ingredients in a portion of the intestine. Examples of encapsulating agents that may be used include polymers and waxes.

[0051] Dosage forms for the topical and / or transdermal application of the compounds described herein may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalers, and / or patches. Typically, the active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier or excipient and / or any desired preservative and / or buffer (where necessary). Additionally, this disclosure covers the use of transdermal patches, which often have the advantage of allowing for increased controlled delivery of the active ingredient into the body. Such dosage forms can be prepared, for example, by dissolving and / or dispensing the active ingredient in a suitable medium. Alternatively or additionally, the rate can be controlled by providing a rate-controlled membrane and / or by dispersing the active ingredient in a polymer matrix and / or gel.

[0052] The fourth aspect of the invention provides the use of the compound of Formula I as described in the first aspect of the invention, or a pharmaceutically acceptable salt thereof, its ester, its deuterated compound, its isotopically labeled derivative, its solvate, its crystalline compound, its stereoisomer, its prodrug, or the pharmaceutical composition described in the third aspect of the invention, in the preparation of a medicament for the prevention or treatment of diseases improved by inhibiting transforming growth factor-β receptor I / ALK5.

[0053] In some embodiments of the present invention, the diseases include gastrointestinal diseases, cancer, fibrotic skin diseases, fibrotic eye diseases, pulmonary fibrosis, or liver fibrosis.

[0054] In some embodiments of the present invention, the gastrointestinal disease is selected from inflammatory bowel disease; in some embodiments of the present invention, the gastrointestinal disease is selected from Crohn's disease and ulcerative colitis.

[0055] In some embodiments of the present invention, the cancer is selected from gastric cancer, esophageal cancer, and colorectal cancer.

[0056] In some embodiments of the present invention, the fibrotic skin disease includes scleroderma, nephrotic fibrotic skin disease, mixed connective tissue disease, sclerosing myxedema, sclerosis, and eosinophilic fasciitis.

[0057] In some embodiments of the present invention, the fibrotic eye disease is selected from dry eye disease, age-related macular degeneration, corneal and conjunctival scarring, postcataract fibrosis, proliferative vitreoretinopathy, or proliferative diabetic retinopathy.

[0058] The beneficial effects of this invention are as follows: The compound represented by Formula I is a novel, highly efficient, organ-restricted ALK5 inhibitor with excellent ALK5 inhibitory activity. Its IC50 value for inhibiting ALK5 activity is 0.879-5.23 nmol / L. It has the characteristics of low systemic exposure risk and has broad potential for development into a treatment for gastrointestinal fibrosis and gastrointestinal tumors (oral administration), pulmonary fibrosis (inhalation administration), and fibrotic skin and eye diseases (topical application). Detailed Implementation

[0059] The following examples provide a more detailed description of the specific implementation of the present invention, but the implementation and protection of the present invention are not limited thereto. It should be noted that any processes not specifically described below are methods that can be implemented or understood by those skilled in the art by referring to existing technology. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0060] Terminology Definition "Substitution" refers to the replacement of hydrogen atoms in a molecule with other different atoms or groups.

[0061] "Multiple" refers to two or more groups. Therefore, the substitution by multiple groups described in this invention means substitution by two or more groups. The specific number of substituents is affected by the number of substituted sites and steric hindrance of the substituted group. It usually means substitution by two, three, four, five or six groups, and more preferably substitution by two or three groups.

[0062] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0063] The term "alkyl" refers to a fully saturated straight-chain or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, linked by a single bond to the remainder of the molecule, having, for example, 1 to 8 (preferably 1 to 6) carbon atoms. In some embodiments, the alkyl group contains 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Non-limiting examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, tert-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, n-octyl, etc. The alkyl groups in the various embodiments of the present invention are preferably C14. 1~6 Alkyl groups. Unless otherwise specifically specified in this specification, alkyl groups may optionally be substituted.

[0064] The term "alkoxy" refers to a group obtained by substituting a hydrogen atom on a hydroxyl group with an alkyl group as defined herein. For example, the term "C 1~8 "Alkoxy" refers to the group -OC 1~8 Alkyl groups, such as methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentoxy, 2-pentoxy, isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, etc.

[0065] The term "heteroaryl" refers to a conjugated cyclic group having carbon atoms (e.g., 3 to 15 carbon atoms, 3 to 14 carbon atoms, 3 to 13 carbon atoms, 3 to 12 carbon atoms, 3 to 11 carbon atoms, 3 to 10 carbon atoms, e.g., 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms) and heteroatoms selected from nitrogen, oxygen, and sulfur (e.g., 2 to 6 heteroatoms, more preferably 2 or 3 heteroatoms). In some embodiments, the heteroaryl group may contain 5, 6, 7, 8, 9, or 10 cyclic atoms, e.g., 5 to 10 cyclic carbon atoms, 5 to 9 cyclic atoms, 5 to 8 cyclic atoms, 5 to 7 cyclic atoms, or 5 to 6 cyclic atoms. Unless otherwise specifically indicated in this specification, the heteroaryl group may be a monocyclic, bicyclic, tricyclic, or more cyclic system and may be fused with a cyclic hydrocarbon group, an aryl group, or a heterocyclic group. The nitrogen, carbon, or sulfur atom in the heteroaryl group may optionally be oxidized, and the nitrogen atom may optionally be quaternized. For the purposes of this invention, the heteroaryl group is preferably a stable 5- to 10-membered aromatic group comprising 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur, or a pentane-hexa- or hexaane-hexa-heteroaryl group comprising 2 or 3 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specifically specified in this specification, the heteroaryl group may optionally be substituted. Examples of heteroaryl groups include, but are not limited to, thiophene, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrazinyl, pyrazinyl, pyridazinyl, benzimidazolyl, benzimorpholinyl, benziisodiazolyl, benzotriazolyl, benzopyrazolyl, imidazopyridyl, pyridomorpholinyl, pyrazolopyridyl, indole, furanyl, pyrroleyl, triazolyl, tetrazolyl, triazinyl, pyridinoneyl, pyrimidinoneyl, pyridazinoneyl, inazinyl, isoindoleyl, indazoleyl, isoindazoleyl, purine, quinolinyl, isoquinolinyl, diazonyl, naphthidyl, quinoxolinyl, pteridinyl, carbazoleyl, carbolinyl, phenanthridine, phenanthridine Phenolic, acridine, phenazinyl, isothiazolyl, benzothiazolyl, benzothiophene, oxatriazolyl, cyclolinyl, quinazolinyl, indene, o-diazaphenanthyl, phenoxazinyl, phenthiazolyl, 4,5,6,7-tetrahydrobenzo[b]thiophene, naphthopyridyl, [1,2,4]triazolo[4,3-b]pyrazinyl, [1,2,4]triazolo[4,3-a]pyrazinyl, [1,2,4]triazolo[4,3-c]pyrimidinyl, [1,2,4]triazolo[4,3-a]pyridinyl, imidazo[1,2-a]pyridinyl, imidazo[1,2-b]pyrazinyl, and imidazo[1,2-a]pyrazinyl, etc.

[0066] In this article, "stereoisomer" refers to a compound composed of identical atoms bonded by the same bonds, but with different three-dimensional structures. This invention covers various stereoisomers and mixtures thereof.

[0067] The compounds of the present invention, or pharmaceutically acceptable salts thereof, may contain one or more chiral carbon atoms, and thus may produce enantiomers, diastereomers, and other stereoisomers. Each chiral carbon atom may be defined as (R)- or (S)- based on stereochemistry. The present invention aims to include all possible isomers, as well as their racemic and optically pure forms. The preparation of the compounds of the present invention may be carried out using racemic, diastereomer, or enantiomer as a starting material or intermediate. Optically active isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography. Conventional techniques for the preparation / separation of individual isomers include chiral synthesis from suitable optically pure precursors, or resolution of racemic mixtures (or racemic mixtures of salts or derivatives) using, for example, chiral high-performance liquid chromatography.

[0068] In this invention, "deuterated compound" refers to a compound in which any hydrogen atom is deuterated.

[0069] The term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. A "pharmaceutically acceptable acid addition salt" is a salt formed with an inorganic or organic acid that retains the bioavailability of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobroms, sulfates, nitrates, and phosphates; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetate, trifluoroacetate, propionate, hexanoate, octanoate, decanoate, undecenoate, glycolate, gluconate, lactate, sebate, adipate, glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate, cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, alginate, ascorbate, salicylate, 4-aminosalicylic acid, and naphthalenedisulfonate. These salts can be prepared using methods known in this field.

[0070] "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that retain the bioavailability of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, the following: primary amines, secondary amines, and tertiary amines; substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in the art.

[0071] As used herein, the terms “crystal form,” “crystalline form,” and “polymorph” are used interchangeably and refer to the crystal structure in which a compound (or its salts, solvates, or other derivatives such as prodrugs or metabolites) can crystallize in different crystalline packings (all having the same elemental composition). Different crystalline forms typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal shapes, optical and electrical properties, stability, and solubility. Recrystallization solvents, crystallization rates, storage temperatures, or other factors can cause one crystalline form to dominate. Polymorphs of a compound can be prepared by crystallization under different conditions.

[0072] The term "solvent" refers to a molecular complex formed by one or more solvent molecules embedded in the crystal lattice of a compound in a stoichiometric or non-stoichiometric ratio, including hydrogen bonding. Common solvents include water, methanol, ethanol, acetic acid, DMSO, THF, diethyl ether, etc. Compounds of Formula I can be prepared, for example, in crystalline form and can be solvates. Suitable solvates include pharmaceutically acceptable solvates and also include both stoichiometric and non-stoichiometric solvates. In some cases, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate can be separated. "Solvent" encompasses both solution phases and separable solvates. Representative solvates include hydrates, ethanolates, and methanolates. The term "hydrate" refers to a compound associated with water. Typically, the number of water molecules in the hydrate of a compound is in a definite ratio to the number of compound molecules in the hydrate. Therefore, a hydrate of a compound can be represented, for example, by the general formula R·xH₂O, where R is the compound and x is a number greater than 0. A given compound can form more than one type of hydrate, including, for example, monohydrates (x is 1), low-hydrates (x is a number greater than 0 and less than 1, such as hemihydrates (R·0.5H2O)), and polyhydrates (x is a number greater than 1, such as dihydrates (R·2H2O), hexahydrates (R·6H2O), etc.).

[0073] The term "prodrug" refers to those compounds that, after administration, will be metabolized (i.e., converted in the body) into the pharmacologically active compounds of the present invention. When the compounds of the present invention are difficult to absorb from the gastrointestinal tract on their own, their bioavailability can be improved by preparing them as prodrugs. Examples of prodrugs of the compounds of the present invention may include simple esters of compounds containing a carboxyl group (e.g., esters obtained by condensation with a C1-4 alcohol according to methods known in the art); esters of compounds containing a hydroxyl group (e.g., esters obtained by condensation with a C1-4 monocarboxylic acid, a C3-6 dicarboxylic acid, or an anhydride thereof, such as succinic anhydride or fumaric anhydride, according to methods known in the art); imines of compounds containing an amino group (e.g., imines obtained by condensation with a C1-4 aldehyde or ketone according to methods known in the art); carbamates of compounds containing an amino group, such as those esters described by Leu et al. (J. Med. Chem., 42: 3623-3628 (1999)) and Greenwald et al. (J. Med. Chem., 42: 3657-3667 (1999)); and aldol acetals or ketal acetals of compounds containing a hydroxyl group (e.g., those acetals obtained by condensation with chloromethyl methyl ether or chloromethyl ethyl ether according to methods known in the art).

[0074] Preparation Example 1: This example provides a method for preparing intermediate 1, wherein the chemical name of intermediate 1 is: 6-[3-(6-methylpyridin-2-yl)-1H-pyrazol-4-yl]benzothiazole, and the structural formula is:

[0075] The specific preparation steps of intermediate 1 are as follows: Step 1: Preparation of 2-methyl-6-(1H-pyrazol-3-yl)pyridine

[0076] 2-Acetyl-6-methylpyridine (20.0 g, 148.0 mmol) and N,N-dimethylformamide dimethyl acetal (150 mL) were added to a 500 mL round-bottom flask, heated to 105 °C, and stirred for 7 hours before stopping the reaction. After cooling, excess N,N-dimethylformamide dimethyl acetal was removed by concentration under reduced pressure to obtain a light brown crude product. The crude product was then slurried with petroleum ether and separated to give an orange solid (20.12 g, 71% yield).

[0077] To the above orange solid (5.97 g, 31.4 mmol), 35 mL of ethanol and 1.80 mL of hydrazine hydrate (36.9 mmol) were added sequentially to obtain a mixture. The mixture was then heated under reflux overnight. After the reaction was complete, it was concentrated, 50 mL of water was added, and the mixture was stirred for 1 hour. The solid was collected by filtration to give compound 2-methyl-6-(1H-pyrazol-3-yl)pyridine (4.25 g, 85% yield). 1 H NMR (CDCl3): δ = 7.64 (d, J = 1.8 Hz, 1H); 7.62 (t, J = 7.6 Hz, 1H); 7.51(d, J = 7.6 Hz, 1H); 7.09 (d, J = 7.7 Hz, 1H); 6.75 (d, J = 2.1 Hz, 1H); 2.57(s, 3H).

[0078] Step 2: Preparation of 2-(4-iodo-1H-pyrazol-3-yl)-6-methylpyridine

[0079] Under nitrogen protection, 2-methyl-6-(1H-pyrazol-3-yl)pyridine (3.3 g, 20.7 mmol) and 25 mL of dry N,N-dimethylformamide were mixed and cooled to 0 °C. Then, N-iodosuccinimide was added in portions, and the mixture was brought to room temperature. Then, it was heated to 90 °C and stirred overnight. After the reaction was complete, ice water and saturated sodium thiosulfate were added, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography to obtain the compound 2-(4-iodo-1H-pyrazol-3-yl)-6-methylpyridine (3.77 g, yield 67%). 1 H NMR (CDCl3): δ = 8.17 (d, J = 8.0 Hz, 1H); 7.69 (m, 2H); 7.15 (d, J =8.0 Hz, 1H); 2.57 (s, 3H).

[0080] Step 3: Preparation of 4-iodo-N,N-dimethyl-3-(6-methylpyridin-2-yl)-1H-pyrazole-1-sulfonamide

[0081] 2-(4-iodo-1H-pyrazol-3-yl)-6-methylpyridine (2.46 g, 8.6 mmol) was dissolved in 100 mL of chloroform, and triethylamine (7 mL, 50 mmol) was added. The mixture was cooled to 0 °C, and then N,N-dimethylaminosulfonyl chloride (4.9 mL, 45.4 mmol) was slowly added dropwise. The mixture was brought to room temperature and heated under reflux overnight. After the reaction was complete, the reaction solution was washed twice with 1 mol / L NaOH solution, the organic phase was dried, filtered, concentrated, and purified by silica gel column chromatography to obtain 4-iodo-N,N-dimethyl-3-(6-methylpyridin-2-yl)-1H-pyrazol-1-sulfonamide (1.76 g, 52% yield). 1 H NMR (CDCl3): δ = 8.09 (s, 1H); 7.72 (d, J = 7.6Hz, 1H); 7.64 (t, J = 8.0 Hz, 1H); 7.17 (d, J = 7.6 Hz, 1H); 2.97 (s, 6H); 2.59 (s, 3H).

[0082] Step 4: Preparation of [1-(N,N-dimethylaminesulfonyl)-3-(6-methylpyridin-2-yl)-pyrazol-4-yl]boronic acid

[0083] Under nitrogen protection, 10.59 g (27 mmol) of 4-iodo-N,N-dimethyl-3-(6-methylpyridin-2-yl)-1H-pyrazole-1-sulfonamide was dissolved in 200 mL of dry tetrahydrofuran and cooled to 0 °C. Then, 32 mL (32 mmol) of 1 mol / L isopropyl magnesium bromide was slowly added dropwise. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Then, cool to 0°C and slowly add trimethyl borate (6 mL, 54 mmol). After the addition is complete, slowly raise the temperature to room temperature and stir for 2 hours. Quench the solution with 100 mL of saturated ammonium chloride, then add 300 mL of 1 mol / L sodium hydroxide to adjust the pH to around 10. Separate the aqueous layer and extract each layer again with 1 mol / L sodium hydroxide. Adjust the pH of the organic layer to 5-6 with acetic acid to precipitate the solid. Cool to 0°C and stir for 30 minutes. Filter to obtain the compound [1-(N,N-dimethylaminesulfonyl)-3-(6-methylpyridin-2-yl)-1H-pyrazol-4-yl]boronic acid (3.85 g, yield 46%). 1 H NMR(CDCl3): δ = 9.15 (bs, 2H); 8.41 (s, 1H); 8.16 (d, J = 8.0 Hz, 1H); 7.76 (t,J = 8.0 Hz, 1H); 7.22 (d, J = 8.0 Hz, 1H); 2.99 (s, 6H); 2.65 (s, 3H).

[0084] Step 5: Preparation of 6-[3-(6-methylpyridin-2-yl)-1H-pyrazol-4-yl]benzothiazole

[0085] Under argon protection, 6-bromobenzothiazole (640 mg, 3.0 mmol), [1-(N,N-dimethylaminesulfonyl)-3-(6-methylpyridin-2-yl)-pyrazol-4-yl]boric acid (1.4 g, 4.5 mmol), PdCl2(dppf)2 (110 mg, 0.15 mmol), 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl (62 mg, 1.5 mmol), RuPHosPdG2 (116 mg, 0.015 mmol), sodium carbonate (950 mg, 9 mmol), and ethylene glycol dimethyl ether (DME):H2O = 5:1 (12 mL) were added to a 100 mL sealed tube. The tube was purged with argon three times and stirred at 100 °C for 3 h. TLC showed that after the reactants (petroleum ether (PE): ethyl acetate (EA) = 2:1, Rf = 0.5) were reacted, 10 mL of water and 15 mL of EA were added and separated. The mixture was evaporated to dryness and separated by Flash chromatography (PE:EA(v:v) = 2:1) to give 779 mg of brown solid, yield: 65%.

[0086] Under argon protection, the above intermediate (770 mg, 2.27 mmol) and 30 mL of 2 mol / L hydrochloric acid solution were added to a 50 mL single-necked flask and refluxed at 80 °C for 5 h. TLC showed that the reactant was completely reacted (EA = 100%) and the Rf value was around 0.3. Ice-cold saturated sodium carbonate was added, and EA was added for extraction. The mixture was washed with brine and separated by Flash chromatography (dichloromethane (DCM):MeOH (v:v) = 20:1) to obtain 464 mg of off-white solid, with a yield of 70%. 1 H NMR (CDCl3): δ = 9.02 (s, 1H); 8.15 (d, J = 8.5Hz, 1H); 8.02 (s, 1H); 7.70 (s, 1H); 7.58 (m, 1H); 7.41 (t, J = 7.5 Hz, 1H); 7.11 (d, J = 7.5 Hz, 1H); 7.07 (d, J = 7.5 Hz, 1H); 2.58 (s, 3H); MS m / z(ESI): 293.2 [M+H] + .

[0087] Preparation Example 2 This example provides a method for preparing intermediate 2, wherein the chemical name of intermediate 2 is: 6-[3-(4-methylthiazo-2-yl)-1H-pyrazol-4-yl]benzothiazole, and the structural formula is:

[0088] The specific preparation steps are as follows: Step 1: Preparation of 3-(dimethylamino)-1-(4-methylthiazol-2-yl)propenone

[0089] Under argon purging protection, N,N-dimethylformamide dimethyl acetal (DMF-DMA, 2.41 g, 20.26 mmol) and 1-(4-methylthiazol-2-yl)acetone (2 g, 14.17 mmol) were added to a 200 mL single-necked flask. The mixture was heated to 75 °C and stirred for 0.5 h, then heated to 105 °C and refluxed with stirring for 5 h. The solution changed from pale yellow to reddish-brown, and TLC showed that the reactants had completely reacted. After cooling to 5-10 °C, an orange-yellow solid precipitated. The solid was filtered, and the filter cake was washed with methyl tert-butyl ether. The filtrate was concentrated and passed through a silica gel column (PE:EA(v:v) = 1:1). The filtrates were combined to give 2.32 g of orange-yellow solid, with a yield of 83%.

[0090] Step 2: Preparation of 4-methyl-2-(1H-pyrazol-3-yl)thiazole

[0091] Under argon purging protection, the first-step intermediate [(E)-3-(dimethylamino)-1-(4-methylthiazolyl-2-yl)prop-2-en-1-one] (2.32 g, 11.82 mmol) and 10 mL of anhydrous ethanol were added to a 100 mL single-necked flask. Hydrazine hydrate (2.51 mL, 41.37 mmol) was added at 10 °C. The mixture was heated to 60 °C and stirred for 5 h. The solution changed from brownish-red to light yellow. TLC showed that the starting material had reacted completely. 50 mL of water and 20 mL of EA were added for extraction. The organic layer was washed with brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 1.83 g of light yellow solid, with a yield of 94%.

[0092] Step 3: Preparation of 2-(4-iodo-1H-pyrazol-3-yl)-4-methylthiazole

[0093] Under argon purging protection, the intermediate from step two, [4-methyl-2-(1H-pyrazol-3-yl)thiazole] (1.8 g, 10.89 mol), and 10 mL of anhydrous DMF were added to a 200 mL single-necked flask. N-iodosuccinimide (NIS, 2.94 g, 13.07 mmol) was added at 0 °C, and the mixture was heated to 90 °C and stirred for 5 h. The sample was quenched with saturated sodium thiosulfate and sodium bicarbonate, extracted with EA, and TLC showed that the starting material had reacted completely. The reaction flask was cooled, and the mixture was poured into 100 mL of stirred ice water. 25 mL of saturated sodium thiosulfate solution and 25 mL of saturated sodium carbonate solution were added, and a yellow solid precipitated. After filtration, the filter cake was washed with water and petroleum ether, and then evaporated to dryness at 45 °C to obtain 3.2 g of yellow solid.

[0094] Step 4: Preparation of 4-iodo-N,N-dimethyl-3-(4-methylthiazolyl-2-yl)-1H-pyrazole-1-sulfonamide

[0095] Under argon protection, the intermediate from step three (3.2 g, 11 mmol) was dissolved in DMF (50 mL), and 0.53 g (22 mmol) of 60% sodium hydride was added. The mixture was cooled to 0 °C, and then N,N-dimethylaminosulfonyl chloride (2.4 mL, 22 mmol) was slowly added dropwise. The mixture was brought to room temperature and stirred for 5 h. After the reaction was complete, the reaction solution was poured into ice water, extracted three times with methyl ether, washed twice with semi-saturated brine, dried the organic phase, filtered and concentrated, and then subjected to silica gel column chromatography (PE:EA(v:v) = 4:1) to give 3.427 g of a pale yellow solid, with a yield of 78%.

[0096] Step 5: Preparation of [1-(N,N-dimethylaminesulfonyl)-3-(4-methylthiazolyl-2-yl)-pyrazole-4-yl]boronic acid

[0097] Under nitrogen protection, the intermediate from step four (3.42 g, 8.59 mmol) was dissolved in 60 mL of dry tetrahydrofuran and cooled to 0 °C. Then, 1 mol / L isopropyl magnesium bromide (10.3 mL, 10.31 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Then, the mixture was cooled to 0 °C and trimethyl borate (1.92 mL, 17.18 mmol) was slowly added dropwise. After the addition was complete, the mixture was slowly heated to room temperature and stirred for 2 hours. 30 mL of saturated ammonium chloride was added to quench the reaction, followed by 100 mL of 1 mol / L sodium hydroxide. The pH was adjusted to approximately 10. The aqueous layer was separated, and the organic layer was extracted once again with 1 mol / L sodium hydroxide. The pH of the organic layer was adjusted to 5-6 with acetic acid, stirred for 10 min, and then evaporated to dryness. The aqueous layer was adjusted to pH 5-6 with hydrochloric acid, and extracted with DCM:MeOH (v:v) = 15:1. The combined organic layers were evaporated to dryness and column chromatography to obtain 1.45 g of a pale yellow solid, with a yield of 53%.

[0098] Step 6: Preparation of 6-[3-(4-methylthiazo-2-yl)-1H-pyrazol-4-yl]benzothiazole

[0099] Under argon protection, 6-bromobenzothiazole (320 mg, 1.5 mmol), [1-(N,N-dimethylaminesulfonyl)-3-(4-methylthiazolyl)-pyrazol-4-yl]boric acid (708 mg, 2.25 mmol), PdCl2(dppf)2 (55 mg, 0.075 mmol), 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl (31 mg, 0.75 mmol), RuPHosPdG2 (58 mg, 0.075 mmol), sodium carbonate (475 mg, 4.5 mmol), DME:H2O = 5:1 (2 mL) were added to a 25 mL sealed tube. The tube was purged with argon three times and stirred at 100 °C for 3 h. TLC showed that after the raw material reacted completely (PE:EA(v:v)=2:1, Rf=0.5), its polarity was similar to that of the boric acid raw material. 5 mL of water and 5 mL of EA were added and separated. After rotary evaporation and Flash chromatography (PE:EA(v:v)=2:1), 0.68 g of brownish-yellow solid was obtained, which was the sulfonamide raw material.

[0100] Under argon protection, sulfonamide raw material (580 mg, 1.43 mmol) and 5 mL of 2 mol / L hydrochloric acid solution were added to a 50 mL single-necked flask. The mixture was refluxed at 80 °C for 5 h. TLC showed that the raw material had reacted completely (EA = 100%), with an Rf value of 0.3. The mixture was then added to ice-cold saturated sodium carbonate, extracted with EA, washed with brine, and separated by Flash chromatography (DCM:MeOH (v:v) = 20:1) to obtain a light yellow solid. The solid was slurried with 10 mL of DCM and 20 mL of n-heptane, and filtered to obtain 0.248 g of a white solid, with a yield of 58%. 1 H NMR (500 MHz, Chloroform- d ) δ 11.89 (s, 1H), 9.04 (d, J = 1.9 Hz, 1H), 8.18 (d, J = 8.3 Hz,1H), 8.14 (s, 1H), 7.74 (s, 1H), 7.64 (d, J = 8.4 Hz, 1H), 6.82 (s, 1H), 2.48 (s, 3H); MS m / z (ESI): 299.3 [M+H] + .

[0101] Preparation Example 3 This example provides a method for preparing intermediate 3, wherein the chemical name of intermediate 3 is: 2-[3-(6-methylpyridin-2-yl)-1H-pyrazol-4-yl]naphthidine, and the structural formula is:

[0102] The specific reaction formula and preparation steps are as follows:

[0103] Under argon protection, 2-chloronaphthidine (500 mg, 0.304 mmol), boric acid (142 mg, 0.456 mmol), PdCl2(dppf)2 (11 mg, 0.0152 mmol), 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl (6 mg, 0.152 mmol), RuPHosPdG2 (12 mg, 0.0152 mmol), sodium carbonate (97 mg, 0.912 mmol), and DME:H2O = 5:1 (2.4 mL) were added to a 25 mL sealed tube. The tube was purged with argon three times and stirred at 100 °C for 3 h. TLC showed that the reactants had largely reacted. 5 mL of water and 5 mL of EA were added, and the mixture was separated by rotary evaporation and Flash chromatography (EA / PE = 1 / 4) to obtain 70 mg of a brownish-yellow solid, with a yield of 68%.

[0104] Under argon protection, 70 mg of the prepared brownish-yellow solid raw material and 2 mL of hydrochloric acid were added to a 100 mL single-necked flask. The mixture was reacted at 80 °C for 5 h. TLC showed that the raw material had reacted completely. Saturated sodium bicarbonate was added to adjust the pH to 7-8, and DCM:MeOH (v:v) = 15:1 (10 mL) was added for extraction. After washing with brine, the mixture was evaporated to dryness and column filtered (DCM:MeOH (v:v) = 10:1) to obtain 50 mg of a yellow oily substance. MS m / z (ESI): 288.2 [M+H] + .

[0105] Preparation Example 4 This example provides a method for preparing intermediate 4, wherein the chemical name of intermediate 4 is: [3-(4-methylthiazolyl-2-yl)-1H-pyrazole-4-yl]naphthidine, and the structural formula is:

[0106] The specific reaction formula and reaction steps are as follows:

[0107] Under argon protection, 2-chloronaphthylidine (150 mg, 0.911 mmol), boric acid (432 mg, 1.37 mmol), PdCl2(dppf)2 (33 mg, 0.046 mmol), 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl (19 mg, 0.046 mmol), RuPHosPdG2 (36 mg, 0.046 mmol), sodium carbonate (290 mg, 0.73 mmol), DME:H2O = 5:1 (2 mL) were added to a 25 mL sealed tube. The mixture was purged with argon three times and stirred at 100 °C for 6 h. After TLC showed complete reaction of the raw materials, 5 mL of water and 5 mL of EA were added, and the mixture was separated by rotary evaporation and Flash chromatography (PE:EA(v:v) = 2:1) to obtain 450 mg of a brown solid, which was the sulfonamide raw material. 1 HNMR (500 MHz, deuterated chloroform) δ 8.96 (dd, J = 4.2, 1.6 Hz, 1H), 8.65 (s, 1H), 8.44 –8.28 (m, 3H), 7.64 (dd, J = 8.6, 4.1 Hz, 1H), 7.05 – 6.86 (m, 1H), 3.09 (s,6H), 2.46 (d, J = 1.0 Hz, 3H).

[0108] Under argon protection, sulfonamide raw material (0.45 g, 1.12 mmol) and 4 mol / L hydrochloric acid (5.6 mL, 22.4 mmol) were added to a 100 mL single-necked flask. The mixture was stirred at 80 °C for 3 h. TLC showed complete reaction of the raw material. Ice and 20 mL of EA were added, and the pH was adjusted to 7-8 with 2 mol / L sodium hydroxide solution. The mixture was washed with saturated brine and dried over anhydrous sodium sulfate to obtain 250 mg of a brown solid, which was intermediate 4. MS m / z (ESI): 294.1 [M+H] + .

[0109] Preparation Example 5 This example provides a method for preparing intermediate 5, the structural formula of which is as follows:

[0110] The reaction formula and specific steps are as follows:

[0111] Under argon protection, the following reactants were added to a 15 mL sealed tube: bromine reactant (180 mg, 0.861 mmol), boric acid reactant (220 mg, 0.889 mmol), PdCl₂(dppf)₂ (38 mg, 0.052 mmol), 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl (22 mg, 0.052 mmol), RuPHosPdG₂ (40 mg, 0.052 mmol), and sodium carbonate (274 mg, 2.58 mmol). The ratio of DME to H₂O was 5:1 (2.5 mL / 0.5 mL). The tube was purged with argon three times and stirred at 100 °C for 3 h. TLC showed complete reaction of the reactants. Silica gel was added, the mixture was evaporated to dryness, and separated by Flash chromatography with an eluent ratio of PE:EA (v:v) of 1:1, yielding 230 mg of a pale yellow oil (84% yield).

[0112] Under argon protection, the above-mentioned pale yellow oily raw material (230 mg, 0.574 mmol) and 2 mol / L hydrochloric acid solution (6.0 mL, 11.5 mmol) were added to a 50 mL single-necked flask. The mixture was stirred at 80 °C for 5 h, and TLC showed that the raw material did not react completely. Hydrochloric acid (1.0 mL) was added, and the mixture was stirred at 80 °C for 1 h, after which the raw material reacted completely. The mixture was extracted twice with methyl tert-butyl ether (MTBE), and the organic phase was discarded. The aqueous layer was adjusted to pH 7-8 with sodium bicarbonate solution, and extracted twice with DCM:MeOH (v:v) = 15:1. The organic phases were combined, washed with saturated brine, dried, filtered, and evaporated to dryness to obtain 146 mg of a brown solid, with a yield of 87%. MS m / z (ESI): 294.2 [M+H] + .

[0113] Preparation Example 6 This example provides a method for preparing intermediate 6, the structural formula of which is:

[0114] The reaction formula and specific preparation steps are as follows:

[0115] Under argon protection, the following ingredients were added to a 15 mL sealed tube: brominated feedstock (250 mg, 1.16 mmol), boric acid feedstock (289 mg, 0.93 mmol), PdCl2(dppf)2 (51 mg, 0.069 mmol), 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl (29 mg, 0.069 mmol), RuPHosPdG2 (54 mg, 0.069 mmol), sodium carbonate (370 mg, 3.49 mmol), and DME:H2O = 5:1 (2.5 mL / 0.5 mL). The tube was purged with argon three times and stirred at 100 °C for 3 h. TLC showed that the reaction was complete, and a new spot was formed (PE:EA(v:v) = 2:1, Rf = 0.5). Silica gel was added, the mixture was evaporated to dryness, and then separated by Flash chromatography with eluent:PE:EA (v:v)=2:1, yielding 288 mg of foamy solid, with a yield of 78%.

[0116] Under argon protection, a foamy solid feedstock (288 mg, 0.719 mmol) and a 2 mol / L hydrochloric acid solution (8.0 mL, 14.38 mmol) were added to a 50 mL single-necked flask. The mixture was stirred at 80 °C for 5 h, and TLC showed that the feedstock reacted completely. The mixture was extracted twice with MTBE, the organic phase was discarded, and the aqueous layer was adjusted to pH 7-8 with sodium bicarbonate solution. DCM:MeOH (v:v) = 15:1 was added for two more extractions. The organic phases were combined, washed with saturated brine, dried, filtered, and evaporated to dryness to obtain 204 mg of a white solid, in equivalence yield. MS m / z (ESI): 294.2 [M+H] + .

[0117] Preparation Example 7 This example provides a method for preparing intermediate 7, the structural formula of which is:

[0118] The reaction formula and specific preparation steps are as follows:

[0119] Under argon protection, the following reactants were added to a 15 mL sealed tube: bromine reactant (226 mg, 1.05 mmol), boric acid reactant (250 mg, 0.806 mmol), PdCl₂(dppf)₂ (30 mg, 0.0403 mmol), 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl (17 mg, 0.0403 mmol), RuPHosPdG₂ (32 mg, 0.0403 mmol), and sodium carbonate (256 mg, 2.42 mmol). The ratio of DME to H₂O was 5:1 (2.5 mL / 0.5 mL). The tube was purged with argon three times and stirred at 100 °C for 3 h. TLC showed complete reaction of the reactants. Silica gel was added, the mixture was evaporated to dryness, and separated by Flash chromatography with an eluent ratio of PE:EA (v:v) of 3:1, yielding 288 mg of a yellow, foamy solid (90% yield).

[0120] Under argon protection, a yellow foamy solid (288 mg, 0.719 mmol), tetrahydrofuran (3.0 mL), and 3 mol / L hydrochloric acid solution (8.0 mL, 21.57 mmol) were added to a 50 mL single-necked flask. The mixture was stirred at 80 °C for 5 h, and TLC showed that the starting material reacted completely. The mixture was extracted twice with MTBE, the organic phase was discarded, and the aqueous layer was adjusted to pH 7-8 with sodium bicarbonate solution. DCM:MeOH (v:v) = 15:1 was added for two more extractions. The organic phases were combined, washed with saturated brine, dried, filtered, and evaporated to dryness to obtain 170 mg of a brown solid, yield 80%. MS m / z (ESI): 294.1 [M+H] + .

[0121] Preparation Example 8 This example provides a method for preparing intermediate 8, the structural formula of which is:

[0122] The reaction formula and specific preparation steps are as follows:

[0123] Under argon protection, the following reactants were added to a 15 mL sealed tube: brominated feedstock (226 mg, 1.12 mmol), boric acid feedstock (250 mg, 0.806 mmol), PdCl2(dppf)2 (30 mg, 0.0403 mmol), 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl (17 mg, 0.0403 mmol), RuPHosPdG2 (32 mg, 0.0403 mmol), and sodium carbonate (256 mg, 2.42 mmol). The DME:H2O ratio was 5:1 (2.5 mL / 0.5 mL). The tube was purged with argon three times and stirred at 100 °C for 3 h. TLC showed complete reaction of the reactants. Post-treatment: Silica gel was added, the mixture was evaporated to dryness, and separated by Flash chromatography with an eluent of PE:EA:DCM (v:v:v) = 60:30:10, yielding 240 mg of foamy solid, 78% yield.

[0124] Under argon protection, the above-mentioned foamy solid feedstock (240 mg, 0.626 mmol), tetrahydrofuran (3.0 mL), and 3 mol / L hydrochloric acid solution (8.0 mL, 18.78 mmol) were added to a 50 mL single-necked flask. The mixture was stirred at 80 °C for 3 h, and TLC showed that the feedstock reacted completely. The mixture was extracted twice with MTBE, the organic phase was discarded, and the aqueous layer was adjusted to pH 7-8 with sodium bicarbonate solution. DCM:MeOH (v:v) = 15:1 was added for two more extractions. The organic phases were combined, washed with saturated brine, dried, filtered, and evaporated to dryness to obtain 138 mg of white solid, yield 80%. MS m / z (ESI): 277.2 [M+H] + .

[0125] Preparation Example 9 This example provides a method for preparing intermediate 9, the structural formula of which is:

[0126] The reaction formula and specific preparation steps are as follows:

[0127] Under argon protection, the following ingredients were added to a 15 mL sealed tube: bromine precursor (152 mg, 0.709 mmol), boric acid precursor (200 mg, 0.645 mmol), PdCl₂(dppf)₂ (24 mg, 0.0322 mmol), 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl (13 mg, 0.0322 mmol), RuPHosPdG₂ (25 mg, 0.0322 mmol), sodium carbonate (205 mg, 1.93 mmol), DME:H₂O = 5:1 (2.5 mL / 0.5 mL). The tube was purged with argon three times and stirred at 100 °C for 3 h. TLC showed complete reaction of the starting material. Silica gel was added, the mixture was evaporated to dryness, and separated by Flash chromatography with eluent: PE:EA:DCM (v:v:v) = 60:30:10, yielding 240 mg of foamy solid, 92% yield.

[0128] Under argon protection, the above-mentioned foamy solid feedstock (240 mg, 0.6 mmol), tetrahydrofuran (3.0 mL), and 3 mol / L hydrochloric acid solution (8.0 mL, 18 mmol) were added to a 50 mL single-necked flask. The mixture was stirred at 80 °C for 3 h, and TLC showed that the feedstock reacted completely. The mixture was extracted twice with MTBE, the organic phase was discarded, and the aqueous layer was adjusted to pH 7-8 with sodium bicarbonate solution. DCM:MeOH (v:v) = 15:1 was added for two more extractions. The organic phases were combined, washed with saturated brine, dried, filtered, and evaporated to dryness to obtain 130 mg of white solid, yield 80%. MS m / z (ESI): 293.1 [M+H] + .

[0129] Preparation Example 10 This example provides a method for preparing intermediate 10, the structural formula of which is:

[0130] The specific preparation steps are as follows: Step 1: Preparation of 2-(4-iodo-1H-pyrazol-3-yl)pyridine

[0131] Under argon protection, 2-(1H-pyrazol-3-yl)pyridine (2.5 g, 17.22 mmol) and 20 mL of dry N,N-dimethylformamide were cooled to 0 °C, and N-iodosuccinimide (4.65 g, 20.67 mmol) was added in portions. The mixture was brought to room temperature and then heated to 90 °C with stirring for 5 hours. After cooling, ice water, saturated sodium thiosulfate, sodium carbonate aqueous solution, and n-hexane were added sequentially. A solid precipitated, which was filtered, washed with n-hexane, and evaporated to dryness to obtain 4.5 g of crude product, which was directly used for the next reaction.

[0132] Step 2: Preparation of 4-iodo-N,N-dimethyl-3-(pyridin-2-yl)-1H-pyrazole-1-sulfonamide

[0133] Under argon protection, iodide (4.5 g, 16.6 mmol) and DMF (40 mL) were added to a 100 mL single-necked flask. Sodium hydride (1.33 g, 33.2 mmol) was added in an ice bath, and the mixture was stirred for 0.5 h. Dimethylamine sulfonyl chloride (3.6 mL, 33.2 mmol) was then added, and the mixture was slowly heated to room temperature for 5 h. TLC showed that the starting material had completely reacted. The mixture was poured into ice water, extracted with EA, washed three times with semi-saturated brine, evaporated to dryness, and column-sected (PE:EA(v:v) = 2:1) to obtain a white solid containing impurities. The solid was incubated overnight with 40 mL of PE / EA(v / v) = 10 / 1 to obtain 5 g of white solid, with a yield of 80%.

[0134] 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.79 – 8.70 (m, ¹H), 8.12 (d, J = 1.1 Hz, 1H), 7.95 (dd, J = 8.0, 1.4 Hz, 1H), 7.82 – 7.73 (m, 1H), 7.33 (ddd, J = 7.6,4.9, 1.2 Hz, 1H), 2.99 (d, J = 1.1 Hz, 6H).

[0135] Step 3: Preparation of [1-(N,N-dimethylaminesulfonyl)-3-(pyridin-2-yl)-pyrazol-4-yl]boronic acid

[0136] Under argon protection, the iodinated compound (2.0 g, 5.29 mmol) was dissolved in 20 mL of dry tetrahydrofuran and cooled to 0 °C. Then, 1 mol / L isopropyl magnesium bromide (6.35 mL, 6.35 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Then, the mixture was cooled to 0 °C and trimethyl borate (1.2 mL, 10.58 mmol) was slowly added dropwise. After the addition was complete, the mixture was slowly warmed to room temperature and stirred for 2 hours. The mixture was quenched with 10 mL of saturated ammonium chloride, followed by 30 mL of 1 mol / L sodium hydroxide. The aqueous layer was separated, and the organic layer was extracted once again with 1 mol / L sodium hydroxide. The organic phase was discarded, and the pH of the aqueous layer was adjusted to 5-6 with hydrochloric acid. The mixture was extracted three times with DCM:MeOH (v:v) = 15:1. The organic layers were combined, dried, filtered, and evaporated to dryness to give 1.18 g of white solid, yield 75%.

[0137] Step 4: Preparation of 6-[3-(pyridin-2-yl)-1H-pyrazol-4-yl]benzothiazole

[0138] Under argon protection, 6-bromobenzothiazole (304 mg, 1.42 mmol), [1-(N,N-dimethylaminesulfonyl)-3-(pyridin-2-yl)-pyrazol-4-yl]boric acid (350 mg, 1.18 mmol), PdCl2(dppf)2 (43 mg, 0.0591 mmol), 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-biphenyl (24 mg, 0.0591 mmol), RuPHosPdG2 (46 mg, 0.0591 mmol), sodium carbonate (376 mg, 3.55 mmol), DME:H2O = 5:1 (2.5 mL / 0.5 mL) were added to a 15 mL sealed tube. The tube was purged with argon three times and stirred at 100 °C for 3 h. TLC showed that the starting material had completely reacted. Add silica gel, evaporate to dryness, and separate by Flash chromatography with eluent: PE:EA (v:v) = 1:1 to obtain 305 mg of yellow foamy solid, yield 67%.

[0139] Under argon protection, the above-mentioned yellow foamy solid feedstock (300 mg, 0.778 mmol), tetrahydrofuran (3.0 mL), and 3 mol / L hydrochloric acid solution (8.0 mL, 23.35 mmol) were added to a 100 mL single-necked flask. The mixture was stirred at 80 °C for 3 h, and TLC showed that the feedstock reacted completely. The mixture was extracted twice with MTBE, the organic phase was discarded, and the aqueous layer was adjusted to pH 7-8 with sodium bicarbonate solution. DCM:MeOH (v:v) = 15:1 was added for two more extractions. The organic phases were combined, washed with saturated brine, dried, filtered, and evaporated to dryness to obtain 208 mg of yellow foamy solid, with a yield of 95%. MS m / z (ESI): 279.1 [M+H] + .

[0140] Example 1 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(4-methylbenzyl)acetamide, and its structural formula is:

[0141] The reaction formula and specific preparation steps for this compound are as follows:

[0142] Under argon protection, p-methylbenzylamine (1 g, 8.25 mmol), DCM (10 mL), and triethylamine (1.4 mL, 9.9 mmol) were added to a 100 mL single-necked flask. The mixture was cooled to 0 °C, and bromoacetyl bromide (0.86 mL, 9.9 mmol) was added. The mixture was stirred for 1 h, and the reaction was quenched with 5 mL of ice water. The solution was washed with citric acid, sodium bicarbonate, and saturated brine. The solution was evaporated to dryness and column chromatography to give 1.7 g of a white brominated solid, yield 85%. MS m / z (ESI): 242.2 [M+H]. + .

[0143] Under argon protection, intermediate 1 (50 mg, 0.171 mmol), the above-mentioned brominated product (54 mg, 0.222 mmol), potassium carbonate (47 mg, 0.342 mmol), and acetonitrile (3 mL) were added to a 25 mL tubing. The mixture was stirred at 85 °C for 6 h. TLC showed that the reaction was complete. The product was evaporated to dryness and separated by Flash chromatography (DCM:MeOH (v:v) = 50:1) to give 32 mg of a white solid, with a yield of 42%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.97 (s, ¹H), 8.04 (d, ¹H) J = 8.5 Hz, 1H), 7.98 (d, J = 1.7 Hz, 1H),7.69 (s, 1H), 7.53 – 7.44 (m, 2H), 7.22 (d, J = 7.7 Hz, 1H), 7.15 – 7.08 (m,5H), 6.72 (s, 1H), 4.97 (s, 2H), 4.43 (d, J = 5.8 Hz, 2H), 2.51 (s, 3H), 2.31(s, 3H); MS m / z (ESI): 454.2[M+H] + .

[0144] Example 2 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,6-dichlorobenzyl)acetamide, and its structural formula is:

[0145] The reaction formula and specific preparation steps for this compound are as follows:

[0146] Under argon protection, 2,6-dichlorobenzylamine (1 g, 5.6 mmol), DCM (10 mL), and triethylamine (0.95 mL, 6.82 mmol) were added to a 100 mL single-necked flask. The mixture was cooled to 0 °C, and bromoacetyl bromide (0.6 mL, 6.82 mmol) was added. The mixture was stirred for 1 h, and the reaction was quenched with 5 mL of ice water. The mixture was washed with citric acid, sodium bicarbonate, and saturated brine. The solution was evaporated to dryness and passed through a column to obtain 0.72 g of an off-white solid, with a yield of 43%.

[0147] Under argon protection, intermediate 1 (50 mg, 0.171 mmol), the above-mentioned brominated intermediate (66 mg, 0.222 mmol), potassium carbonate (47 mg, 0.342 mmol), and acetonitrile (3 mL) were added to a 25 mL tubing. The mixture was stirred at 85 °C for 6 h. TLC showed that the reaction was complete. The sample was evaporated to dryness and separated by Flash chromatography (DCM:MeOH(v:v) = 100:1 → 30:1) to give 32 mg of a white solid, with a yield of 33%. 1 H NMR (500 MHz, deuterated chloroform) δ 8.97 (s, 1H), 8.04 (d, J = 8.5 Hz, 1H), 7.98 (d, J =1.7 Hz, 1H), 7.66 (s, 1H), 7.50 (t, J = 7.7 Hz, 1H), 7.45 (dd, J = 8.5, 1.7 Hz, 1H), 7.27 (d, J = 8.2 Hz, 2H), 7.23 (d, J = 7.8 Hz, 1H), 7.15 (t, J = 8.0 Hz, 1H), 7.10 (d, J = 7.7 Hz, 1H), 6.96 (t, J = 5.7 Hz, 1H), 4.93 (s, 2H), 4.79 (d, J = 5.8Hz, 2H), 2.50 (s, 3H); MS m / z (ESI):508.2[M+H] + .

[0148] Example 3 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2-trifluoromethylbenzyl)acetamide, and the structural formula is:

[0149] The reaction formula and specific preparation steps for this compound are as follows:

[0150] Under argon protection, o-trifluoromethylbenzylamine (1 g, 5.71 mmol), DCM (10 mL), and triethylamine (1.33 mL, 0.685 mmol) were added to a 100 mL single-necked flask. The mixture was cooled to 0 °C, and bromoacetyl bromide (0.84 mL, 0.685 mmol) was added. The mixture was stirred for 1 h, and the reaction was quenched with 5 mL of ice water. The mixture was washed with citric acid, sodium bicarbonate, and saturated brine. The solution was evaporated to dryness to obtain 1.2 g of an off-white solid, with a yield of 71%.

[0151] Under argon protection, intermediate 1 (50 mg, 0.171 mmol), brominated intermediate (66 mg, 0.222 mmol), potassium carbonate (47 mg, 0.342 mmol), and acetonitrile (3 mL) were added to a 25 mL tubing and stirred at 85 °C for 6 h. TLC showed complete reaction of the starting material. The mixture was then evaporated to dryness and separated by Flash chromatography (DCM:MeOH(v:v) = 100:1 → 40:1) to give 26 mg of a white solid, with a yield of 30%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.4 Hz, 1H), 7.98 (d, J = 1.6Hz, 1H), 7.68 (s, 1H), 7.62 (d, J = 7.8 Hz, 1H), 7.51 (td, J = 7.5, 4.9 Hz, 3H),7.48 – 7.44 (m, 1H), 7.36 (t, J = 7.5 Hz, 1H), 7.23 (d, J = 7.8 Hz, 1H), 7.11 (d, J = 7.6 Hz, 1H), 6.96 (s, 1H), 4.97 (s, 2H), 4.65 (d, J = 6.2 Hz, 2H), 2.51 (s,3H); MS m / z (ESI):508.1[M+H] + .

[0152] Example 4 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2-fluorobenzyl)acetamide, and the structural formula is:

[0153] The reaction formula and specific preparation steps for this compound are as follows:

[0154] Under argon protection, o-fluorobenzylamine (1 g, 7.99 mmol), DCM (10 mL), and triethylamine (1.33 mL, 9.59 mmol) were added to a 100 mL single-necked flask. The mixture was cooled to 0 °C, and bromoacetyl bromide (0.84 mL, 9.59 mmol) was added. The mixture was stirred for 1 h, and the reaction was quenched with 5 mL of ice water. The mixture was washed with citric acid, sodium bicarbonate, and saturated brine. The solution was evaporated to dryness to obtain 1.8 g of an off-white solid, with a yield of 92%.

[0155] Under argon protection, intermediate 1 (50 mg, 1 eq), the aforementioned brominated product (55 mg, 0.222 mmol), potassium carbonate (47 mg, 0.342 mmol), and acetonitrile (3 mL) were added to a 25 mL tubing. The mixture was stirred at 85 °C for 6 h. TLC showed that the reaction was complete. The product was evaporated to dryness and separated by Flash chromatography (DCM:MeOH(v:v) = 100:1 → 40:1) to give 40 mg of a white solid, with a yield of 51%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.97 (s, ¹H), 8.05 (d, ¹H) J = 8.4 Hz, 1H), 7.98 (d, J = 1.7Hz, 1H), 7.68 (s, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.46 (dd, J = 8.5, 1.7 Hz, 1H), 7.30 (td, J = 7.6, 1.8 Hz, 1H), 7.26 – 7.21 (m, 2H), 7.12 – 7.04 (m, 2H), 7.00(ddd, J = 9.7, 8.2, 1.2 Hz, 1H), 6.90 (t, J = 6.2 Hz, 1H), 4.97 (s, 2H), 4.52 (d, J= 6.0 Hz, 2H), 2.51 (s, 3H); MS m / z (ESI): 458.2 [M+H] + .

[0156] Example 5 This example provides a compound with the chemical name: 2-[4-(benzothiazol-6-yl)-3-(4-methylthiazol-2-yl)-1H-pyrazole]-N-(2-methylbenzyl)acetamide, and the structural formula is:

[0157] The reaction formula and specific preparation steps for this compound are as follows:

[0158] Under argon protection, intermediate 2 (50 mg, 0.168 mmol), bromide (61 mg, 0.251 mmol), potassium carbonate (46 mg, 0.336 mmol), and acetonitrile (4 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h, and TLC showed that the starting material had completely reacted. Ice water and DCM were added and separated. The mixture was evaporated to dryness and then separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain a light-colored solid. 1 mL of DCM, 2 mL of n-heptane, and 1 mL of EA were added and the mixture was stirred overnight. After centrifugation, the solid was washed with ice-cold ethanol and centrifuged again to obtain 30 mg of a white solid, with a yield of 39%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 9.01 (s, ¹H), 8.21 (d, ¹H) J = 1.7 Hz, 1H), 8.12(d, J = 8.4 Hz, 1H), 7.69 (s, 1H), 7.62 (dd, J = 8.4, 1.6 Hz, 1H), 7.21 – 7.12(m, 4H), 6.84 (s, 1H), 6.73 (s, 1H), 4.95 (s, 2H), 4.48 (d, J = 5.5 Hz, 2H), 2.43 (s, 3H), 2.29 (s, 3H); MS m / z (ESI): 460.2 [M+H] + .

[0159] Example 6 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2-methylbenzyl)acetamide, and the structural formula is:

[0160] The reaction formula and specific preparation steps for this compound are as follows:

[0161] Under argon protection, o-methylbenzylamine (3.45 g, 28.47 mmol) and 60 mL of THF were added to a 200 mL single-necked flask. Acetyl bromoacetate (2.6 mL, 29.9 mmol) was added at 0 °C, and the mixture was stirred at room temperature for 24 h. TLC showed that the reaction was essentially complete. Ice water and saturated sodium bicarbonate solution were added, followed by extraction with EA. After washing with saturated brine, the mixture was evaporated to dryness and passed through a silica gel column (PE:EA(v:v) = 10:1 → 2:1) to obtain a white solid, i.e., the brominated intermediate, 2.98 g, with a yield of 43%.

[0162] Under argon protection, intermediate 1 (125 mg, 0.342 mmol), the above-mentioned brominated intermediate (124 mg, 0.513 mmol), potassium carbonate (95 mg, 0.684 mmol), and acetonitrile (10 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h. TLC showed that the starting material had reacted completely. Ice water and DCM were added and separated. The mixture was evaporated to dryness to obtain 60 mg of light-colored solid. 1 mL of DCM and 4 mL of n-heptane were added and the mixture was stirred. After centrifugation, 50 mg of white solid was obtained, with a yield of 30%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.4 Hz, 1H), 7.98 (d, J = 1.7 Hz, 1H), 7.70 (s, 1H), 7.49 (t, J = 7.7 Hz, 1H), 7.45 (dd, J = 8.5, 1.8 Hz, 1H), 7.20 – 7.15 (m, 3H), 7.13 (d, J = 7.5 Hz, 2H), 7.10 (d, J = 7.8 Hz, 1H), 6.76 (s, 1H), 4.98 (s, 2H), 4.47 (d, J = 5.6 Hz, 2H), 2.51 (s, 3H), 2.27 (s, 3H); MS m / z (ESI): 454.2 [M+H] + .

[0163] Example 7 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-benzylacetamide, and the structural formula is:

[0164] The reaction formula and specific preparation steps for this compound are as follows:

[0165] Under argon protection, benzylamine (1 g, 9.33 mmol), 20 mL of DCM, and triethylamine (1.32 mL, 9.52 mmol) were added to a 200 mL single-necked flask. Bromoacetyl bromide (0.83 mL, 9.52 mmol) was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water (EA), washed with saturated brine, washed with 5% citric acid solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness before column chromatography to give 1.3 g of a white solid, the brominated intermediate, in a yield of 61%.

[0166] Under argon protection, intermediate 1 (125 mg, 0.342 mmol), the above-mentioned brominated intermediate (117 mg, 0.513 mmol), potassium carbonate (95 mg, 0.684 mmol), and acetonitrile (10 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h. TLC showed that the starting material had completely reacted. Ice water and DCM were added and separated. The mixture was evaporated to dryness to obtain a light-colored solid. 4 mL of DCM, 4 mL of n-heptane, and 1.5 mL of EA were added and the mixture was stirred overnight and filtered. The mixture was washed once with ice-cold ethanol and evaporated to dryness to obtain 28 mg of white solid, with a yield of 19%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.5 Hz, 1H), 7.98 (d, J = 1.7 Hz, 1H), 7.70(s, 1H), 7.50 (t, J = 7.7 Hz, 1H), 7.45 (dd, J = 8.5, 1.7 Hz, 1H), 7.31 – 7.27(m, 2H), 7.25 – 7.20 (m, 4H), 7.11 (d, J = 7.6 Hz, 1H), 6.80 (s, 1H), 4.99 (s,2H), 4.48 (d, J = 5.9 Hz, 2H), 2.51 (s, 3H); MS m / z (ESI): 440.2 [M+H]+ .

[0167] Example 8 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2-chlorobenzyl)acetamide, and the structural formula is:

[0168] The reaction formula and specific preparation steps for this compound are as follows:

[0169] Under argon protection, o-chlorobenzylamine (4 g, 28.25 mmol), 60 mL of DCM, and triethylamine (4 mL, 28.81 mmol) were added to a 200 mL single-necked flask. Acetyl bromoacetate (2.5 mL, 28.81 mmol) was added at -5 °C, and the mixture was stirred at room temperature for 10 min. TLC showed that the reaction was essentially complete. The mixture was extracted with ice water (EA), washed with saturated brine, washed with 5% citric acid solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to give 6.8 g of a white solid, the brominated intermediate, with a yield of 92%.

[0170] Under argon protection, pyrazole starting material (125 mg, 0.342 mmol), bromide (135 mg, 0.513 mmol), potassium carbonate (95 mg, 0.684 mmol), and acetonitrile (10 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h. TLC showed that the starting material had completely reacted. Ice water was added, and the mixture was separated from DCM and evaporated to dryness to obtain a light-colored solid. 4 mL of DCM, 4 mL of n-heptane, and 1.5 mL of EA were added, and the mixture was stirred overnight. Washing with ice-cold ethanol gave 66 mg of a white solid, with a yield of 42%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.5 Hz, 1H), 7.99 (d, J = 1.7 Hz, 1H), 7.68 (s, 1H), 7.51 (t, J =7.8 Hz, 1H), 7.46 (dd, J = 8.5, 1.7 Hz, 1H), 7.36 – 7.31 (m, 2H), 7.25 (d, J =7.6 Hz, 1H), 7.23 – 7.19 (m, 2H), 7.11 (d, J = 7.6 Hz, 1H), 7.00 (t, J= 6.2 Hz,1H), 4.97 (s, 2H), 4.55 (d, J = 6.2 Hz, 2H), 2.52 (s, 3H); MS m / z (ESI): 474.1 [M+H] + .

[0171] Example 9 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetamide, and the structural formula is:

[0172] The reaction formula and specific preparation steps for this compound are as follows:

[0173] Under argon protection, 2,6-difluorobenzylamine (4 g, 6.99 mmol), 60 mL of DCM, and triethylamine (4 mL, 7.13 mmol) were added to a 200 mL single-necked flask. Acetyl bromobromine (2.5 mL, 7.13 mmol) was added at -5 °C, and the mixture was stirred at room temperature for 10 min. TLC showed that the reaction was essentially complete. The mixture was extracted with ice water (EA), washed with saturated brine, washed with 5% citric acid solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to give 1.6 g of a white solid, yield 87%.

[0174] Under argon protection, intermediate 1 (125 mg, 0.342 mmol), the brominated product obtained above (135 mg, 0.513 mmol), potassium carbonate (95 mg, 0.684 mmol), and acetonitrile (10 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h. TLC showed that the starting material had reacted completely. Ice water and DCM were added and separated. The mixture was evaporated to dryness and separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain 70 mg of light-colored solid. 4 mL of DCM, 4 mL of n-heptane, and 1.5 mL of EA were added and stirred overnight. After filtration, the mixture was washed with ice-cold ethanol to obtain 58 mg of white solid, with a yield of 36%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.96 (s, ¹H), 8.03 (d, ¹H) J = 8.5Hz, 1H), 7.96 (d, J = 1.7 Hz, 1H), 7.66 (s, 1H), 7.50 (t, J = 7.7 Hz, 1H), 7.44 (dd, J= 8.5, 1.7 Hz, 1H), 7.25 – 7.17 (m, 2H), 7.09 (d, J = 7.6 Hz, 1H), 7.00 –6.93 (m, 1H), 6.84 (t, J = 7.8 Hz, 2H), 4.93 (s, 2H), 4.57 (d, J = 5.8 Hz, 2H),2.50 (s, 3H); MS m / z (ESI): 476.2 [M+H] + .

[0175] Example 10 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2-methylbenzyl)acetamide, and the structural formula is:

[0176] The reaction formula and specific preparation steps for this compound are as follows:

[0177] Under argon protection, intermediate 3 (50 mg, 0.174 mmol), bromide (63 mg, 0.261 mmol), potassium carbonate (48 mg, 0.348 mmol), and acetonitrile (8 mL) were added to a 25 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h, and TLC showed that the starting material had completely reacted. Ice water and DCM were added and separated. The mixture was evaporated to dryness and then separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain a light-colored solid. 1 mL of DCM, 2 mL of n-heptane, and 1 mL of EA were added and the mixture was stirred overnight. After centrifugation, the solid was washed with 1 mL of pre-cooled ethanol and centrifuged again to obtain 31 mg of a light yellow solid, with a yield of 40%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.92 (dd, J = 4.2, 1.6 Hz, 1H), 8.33(d, J = 8.5 Hz, 1H), 8.25 – 8.20 (m, 2H), 7.81 (s, 1H), 7.65 – 7.59 (m, 2H),7.45 (d, J = 7.7 Hz, 1H), 7.20 (d, J = 7.3 Hz, 1H), 7.18 – 7.10 (m, 4H), 6.92 (s,1H), 5.00 (s, 2H), 4.48 (d,J = 5.6 Hz, 2H), 2.50 (s, 3H), 2.28 (s, 3H); MS m / z(ESI):449.2 [M+H] + .

[0178] Example 11 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-benzylacetamide, and the structural formula is:

[0179] The reaction formula and specific preparation steps for this compound are as follows:

[0180] Under argon protection, intermediate 3 (50 mg, 0.174 mmol), bromoacetamide starting material (60 mg, 0.261 mmol), potassium carbonate (48 mg, 0.348 mmol), and acetonitrile (5 mL) were added to a 25 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h, and TLC showed that the starting material had completely reacted. Ice water was added, and DCM was separated. The mixture was evaporated to dryness and then separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain a light-colored solid. 1 mL of DCM, 2 mL of n-heptane, and 1 mL of EA were added, and the mixture was stirred overnight. After centrifugation, the solid was washed with 1 mL of pre-cooled ethanol and centrifuged again to obtain 31 mg of a light yellow solid, with a yield of 41%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.94 (dd, J = 4.2, 1.7 Hz, 1H), 8.34 (dd, J = 8.7, 1.6 Hz, 1H), 8.24 (d, J = 8.8 Hz, 1H), 8.19 (s, 1H), 7.78(dd, J = 8.9, 1.3 Hz, 1H), 7.67 – 7.59 (m, 2H), 7.46 (d, J = 7.7 Hz, 1H), 7.29(ddt, J = 13.0, 9.0, 4.8 Hz, 6H), 7.18 (d, J = 7.7 Hz, 1H), 6.97 (s, 1H), 5.02(s, 2H), 4.50 (d, J= 5.9 Hz, 2H), 2.51 (s, 3H); MS m / z (ESI): 435.1 [M+H] + .

[0181] Example 12 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2-chlorobenzyl)acetamide, and the structural formula is:

[0182] The reaction formula and specific preparation steps for this compound are as follows:

[0183] Under argon protection, intermediate 3 (50 mg, 0.174 mmol), bromoacetamide starting material (68.5 mg, 0.261 mmol), potassium carbonate (48 mg, 0.348 mmol), and acetonitrile (3 mL) were added to a 25 mL single-necked flask. The mixture was refluxed at 60 °C for 3 h, and TLC showed that the starting material had completely reacted. Ice water was added, and DCM was separated. The mixture was evaporated to dryness and then separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain a light-colored solid. 1 mL of DCM, 2 mL of n-heptane, and 1 mL of EA were added, and the mixture was stirred overnight. After centrifugation, the solid was washed with 1 mL of pre-cooled ethanol and centrifuged again to obtain 46 mg of a yellow solid, with a yield of 56%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.92 (dd, J = 4.2, 1.6 Hz, 1H), 8.33 (d, J = 8.5 Hz, 1H), 8.23 ​​(d, J = 8.8 Hz, 1H), 8.16 (s, 1H), 7.80 (d, J = 8.8Hz, 1H), 7.64 – 7.59 (m, 2H), 7.51 (d, J = 7.7 Hz, 1H), 7.38 – 7.29 (m, 2H), 7.20 (dd, J = 5.9, 3.5 Hz, 2H), 7.18 – 7.12 (m, 2H), 4.98 (s, 2H), 4.56 (d, J =6.1 Hz, 2H), 2.49 (s, 3H); MS m / z (ESI):469.2 [M+H] + .

[0184] Example 13 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetamide, and the structural formula is:

[0185] The reaction formula and specific preparation steps for this compound are as follows:

[0186] Under argon protection, intermediate 3 (50 mg, 0.174 mmol), bromoacetamide starting material (69 mg, 0.261 mmol), potassium carbonate (48 mg, 0.348 mmol), and acetonitrile (3 mL) were added to a 25 mL single-necked flask. The mixture was refluxed at 80 °C for 6 h. TLC showed that the starting material had reacted completely, and two new spots were formed. Ice water and DCM were added and separated. The mixture was evaporated to dryness and then separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain a light-colored solid. 1 mL of n-heptane was added, and the mixture was evaporated to dryness to obtain 36 mg of a light yellow solid, with a yield of 44%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.92 (dd, J = 4.1, 1.7 Hz, 1H), 8.32 (ddd, J = 8.5, 1.7, 0.9 Hz, 1H),8.22 (dd, J = 8.8, 0.9 Hz, 1H), 8.14 (s, 1H), 7.80 (d, J = 8.9 Hz, 1H), 7.66 –7.58 (m, 2H), 7.50 (d, J = 7.7 Hz, 1H), 7.25 – 7.17 (m, 1H), 7.16 (d, J = 7.7 Hz,1H), 6.98 (s, 0H), 6.85 (t, J = 7.8 Hz, 2H), 4.94 (s, 2H), 4.59 (d, J = 5.9 Hz, 2H), 2.49 (s, 3H); MS m / z (ESI): 471.3 [M+H] + .

[0187] Example 14 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(4-methylbenzyl)acetamide, and the structural formula is:

[0188] The reaction formula and specific preparation steps for this compound are as follows:

[0189] Under argon protection, intermediate 3 (50 mg, 0.174 mmol), bromoacetamide starting material (63 mg, 0.261 mmol), potassium carbonate (48 mg, 0.348 mmol), and acetonitrile (3 mL) were added to a 25 mL single-necked flask. The mixture was refluxed at 80 °C for 6 h, and TLC showed that the starting material had completely reacted. Ice water was added and DCM was separated. The mixture was evaporated to dryness and then separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain a light-colored solid. 1 mL of n-heptane was added, and the mixture was evaporated to dryness to obtain 43 mg of a light yellow solid, with a yield of 55%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.92 (dd, J = 4.2, 1.7 Hz, 1H), 8.32 (dt, J = 8.5, 1.3 Hz, 1H), 8.22 (dd, J =8.8, 0.9 Hz, 1H), 8.16 (s, 1H), 7.77 (d, J = 8.8 Hz, 1H), 7.65 – 7.57 (m, 2H), 7.44 (d, J = 7.7 Hz, 1H), 7.19 – 7.11 (m, 3H), 7.09 (d, J = 7.8 Hz, 2H), 6.82 (s,1H), 4.98 (s, 2H), 4.43 (d, J = 5.8 Hz, 2H), 2.49 (s, 3H), 2.30 (s, 3H); MS m / z(ESI):449.2 [M+H] + .

[0190] Example 15 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,6-dichlorobenzyl)acetamide, and the structural formula is:

[0191] The reaction formula and specific preparation steps for this compound are as follows:

[0192] Under argon protection, intermediate 3 (50 mg, 0.174 mmol), bromoacetamide starting material (78 mg, 0.261 mmol), potassium carbonate (48 mg, 0.348 mmol), and acetonitrile (3 mL) were added to a 25 mL single-necked flask. The mixture was refluxed at 80 °C for 6 h, and TLC showed that the starting material had completely reacted. Ice water was added and DCM was separated. The mixture was evaporated to dryness and then separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain a light-colored solid. 1 mL of n-heptane was added, and the mixture was evaporated to dryness to obtain 30 mg of a light yellow solid, with a yield of 34%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.92 (dd, J = 4.1, 1.7 Hz, 1H), 8.32 (d, J = 8.5 Hz, 1H), 8.22 (d, J = 8.8 Hz,1H), 8.15 (s, 1H), 7.80 (d, J = 8.8 Hz, 1H), 7.65 – 7.57 (m, 2H), 7.50 (d, J =7.7 Hz, 1H), 7.28 (s, 1H), 7.18 – 7.10 (m, 3H), 4.94 (s, 2H), 4.80 (d, J = 5.8Hz, 2H), 2.49 (s, 3H); MS m / z (ESI):503.1 [M+H] + .

[0193] Example 16 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2-trifluoromethylbenzyl)acetamide, and the structural formula is:

[0194] The reaction formula and specific preparation steps for this compound are as follows:

[0195] Under argon protection, intermediate 3 (50 mg, 0.174 mmol), bromoacetamide starting material (77 mg, 0.261 mmol), potassium carbonate (48 mg, 0.348 mmol), and acetonitrile (3 mL) were added to a 25 mL single-necked flask. The mixture was refluxed at 80 °C for 6 h, and TLC showed that the starting material had completely reacted. Ice water and DCM were added and separated. The mixture was evaporated to dryness and then separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain a light-colored solid. 1 mL of n-heptane was added, and the mixture was evaporated to dryness to obtain 60 mg of a light yellow solid, with a yield of 69%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.92 (dd, J = 4.2, 1.6 Hz, 1H), 8.35 – 8.30 (m, 1H), 8.22 (d, J = 8.8 Hz, 1H),8.16 (s, 1H), 7.78 (d, J = 8.8 Hz, 1H), 7.62 (dd, J = 8.3, 4.4 Hz, 3H), 7.54 (d, J = 7.6 Hz, 1H), 7.49 (t, J = 8.2 Hz, 2H), 7.36 (t, J = 7.6 Hz, 1H), 7.16 (d, J = 7.7Hz, 1H), 7.08 (d, J = 6.9 Hz, 1H), 4.98 (s, 2H), 4.66 (d, J = 6.2 Hz, 2H), 2.48(s, 3H); MS m / z (ESI):503.3 [M+H] + .

[0196] Example 17 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2-fluorobenzyl)acetamide, and the structural formula is:

[0197] The reaction formula and specific preparation steps for this compound are as follows:

[0198] Under argon protection, intermediate 3 (50 mg, 0.174 mmol), bromoacetamide starting material (64 mg, 0.261 mmol), potassium carbonate (48 mg, 0.348 mmol), and acetonitrile (3 mL) were added to a 25 mL single-necked flask. The mixture was refluxed at 80 °C for 6 h, and TLC showed that the starting material had completely reacted. Ice water and DCM were added and separated. The mixture was evaporated to dryness and then separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain a light-colored solid. 1 mL of n-heptane was added, and the mixture was evaporated to dryness to obtain 50 mg of a light yellow solid, with a yield of 54%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.92 (dd, J = 4.2, 1.6 Hz, 1H), 8.34 – 8.31 (m, 1H), 8.22 (d, J = 8.9 Hz, 1H),8.16 (s, 1H), 7.79 (d, J = 8.8 Hz, 1H), 7.65 – 7.60 (m, 2H), 7.49 (d, J = 7.7 Hz,1H), 7.34 – 7.30 (m, 1H), 7.23 (ddd, J = 7.5, 5.4, 1.9 Hz, 1H), 7.16 (d, J = 7.7Hz, 1H), 7.07 (dd, J = 8.0, 6.8 Hz, 1H), 7.03 – 6.95 (m, 2H), 4.98 (s, 2H), 4.53 (d, J = 6.1 Hz, 2H), 2.49 (s, 3H); MS m / z (ESI): 453.2 [M+H] + .

[0199] Example 18 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(4-methylthiazolyl-2-yl)-1H-pyrazole]-N-(2-methylbenzyl)acetamide, and the structural formula is:

[0200] The reaction formula and specific preparation steps for this compound are as follows:

[0201] After purging the 25 mL sealed tube with argon three times, intermediate 4 (70 mg, 0.238 mmol), bromoacetylbenzylamine starting material (116 mg, 0.478 mmol), potassium carbonate (99 mg, 0.716 mmol), and anhydrous acetonitrile (3 mL) were added. The reaction was carried out at 80 °C for 6 h. TLC showed that the starting material had completely reacted. The sample was then evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 20:1) to give 45 mg of a yellow solid, with a yield of 41%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.95 (dd, J = 4.2, 1.6 Hz, 1H), 8.41 (d, J = 8.9 Hz, 1H), 8.36 (dd, J = 8.2,2.1 Hz, 2H), 8.23 ​​(s, 1H), 7.63 (dd, J = 8.5, 4.2 Hz, 1H), 7.22 – 7.19 (m, 1H), 7.15 (td, J = 7.7, 4.2 Hz, 3H), 6.93 (d, J = 1.2 Hz, 1H), 6.71 (s, 1H), 4.97 (s,2H), 4.49 (d, J = 5.6 Hz, 2H), 2.48 (d, J = 1.1 Hz, 3H), 2.30 (s, 3H); MS m / z(ESI):455.2 [M+H] + .

[0202] Example 19 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3-fluorobenzyl)acetamide, and the structural formula is:

[0203] The reaction formula and specific preparation steps for this compound are as follows:

[0204] Under argon protection, 2 g (16 mmol) of 3-fluorobenzylamine, 10 mL of DCM, and 2.3 mL (16.3 mmol) of triethylamine were added to a 200 mL single-necked flask. Chloroacetyl chloride (1.3 mL, 16.3 mmol) was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 3.1 g of a brown oily substance, which was used directly in the next reaction.

[0205] Under argon protection, intermediate 1 (125 mg, 0.427 mmol), the chlorinated product prepared above (129 mg, 0.641 mmol), potassium carbonate (118 mg, 0.855 mmol), and acetonitrile (10 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h. TLC showed that the starting material had completely reacted. The mixture was then evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 100 mg of a light-colored solid, with a yield of 51%. 1 ¹H NMR (500MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.5 Hz, 1H), 7.98 (d, J = 1.7 Hz,1H), 7.71 (s, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.46 (dd, J = 8.5, 1.7 Hz, 1H), 7.23(d, J = 7.7 Hz, 2H), 7.11 (d, J = 7.6 Hz, 1H), 7.01 (d, J = 7.6 Hz, 1H), 6.95 (s,1H), 6.94 – 6.93 (m, 1H), 5.30 (s, 1H), 5.00 (s, 2H), 4.47 (d, J = 6.0 Hz, 2H),2.52 (s, 3H); MS m / z (ESI):458.2 [M+H] + .

[0206] Example 20 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3-chlorobenzyl)acetamide, and the structural formula is:

[0207] The reaction formula and specific preparation steps for this compound are as follows:

[0208] Under argon protection, 1 g of 3-chlorobenzylamine (7.06 mmol), 10 mL of DCM, and 1.0 mL of triethylamine (7.2 mmol) were added to a 100 mL single-necked flask. Chloroacetyl chloride (0.573 mL, 7.2 mmol) was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution and saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 1.51 g of a brown solid, which was used directly in the next reaction.

[0209] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the chlorinated product obtained above (73 mg, 0.444 mmol), potassium carbonate (62 mg, 0.333 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h. TLC showed that the starting material had completely reacted. The product was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 50 mg of white solid, with a yield of 47.6%. 1 ¹H NMR (500MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.4 Hz, 1H), 7.98 (d, J = 1.7 Hz,1H), 7.71 (s, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.46 (dd, J = 8.4, 1.7 Hz, 1H), 7.23– 7.20 (m, 3H), 7.12 (dd, J = 7.3, 4.2 Hz, 2H), 6.93 (s, 1H), 5.30 (s, 1H), 5.00 (s, 2H), 4.45 (d, J = 6.0 Hz, 2H), 2.52 (s, 3H); MS m / z (ESI): 474.2 [M+H] + .

[0210] Example 21 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3-cyanobenzyl)acetamide, and the structural formula is:

[0211] The reaction formula and specific preparation steps for this compound are as follows:

[0212] Under argon protection, 1 g (7.57 mmol) of 3-cyanobenzylamine, 10 mL of DCM, and 1.0 mL (7.72 mmol) of triethylamine were added to a 100 mL single-necked flask. Chloroacetyl chloride (0.573 mL, 7.72 mmol) was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 1.59 g of a light-colored solid, which was used directly in the next reaction.

[0213] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the chlorinated compound (70 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h. TLC showed that a large amount of the starting material was still unreacted. The temperature was then increased to 90 °C and stirred for 5 h. TLC showed that the starting material had completely reacted. The solution was evaporated to dryness and passed through a column chromatography (DCM:MeOH (v:v) = 100:1) to give 40 mg of a white solid, with a yield of 39%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J =8.5 Hz, 1H), 7.97 (d, J = 1.7 Hz, 1H), 7.71 (s, 1H), 7.55 – 7.50 (m, 3H), 7.49(d, J = 8.0 Hz, 1H), 7.44 (dd, J = 8.5, 1.7 Hz, 1H), 7.39 (t, J = 8.0 Hz, 1H), 7.21(dd, J = 10.7, 6.6 Hz, 2H), 7.12 (d, J = 7.7 Hz, 1H), 5.01 (s, 2H), 4.48 (d, J =6.1 Hz, 2H), 2.52 (s, 3H); MS m / z (ESI):465.1 [M+H] + .

[0214] Example 22 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3-carboxylic acid methyl ester benzyl)acetamide, and the structural formula is:

[0215] The reaction formula and specific preparation steps for this compound are as follows:

[0216] Under argon protection, 2 g (12.11 mmol) of 3-cyanobenzylamine, 10 mL of DCM, and 1.720 mL (12.35 mmol) of triethylamine were added to a 100 mL single-necked flask. Then, 1.1 mL (12.35 mmol) of bromoacetyl bromide was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 2.8 g of a brown solid, which was used directly in the next reaction.

[0217] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (95 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h. TLC showed that a large amount of the starting material was still unreacted. The temperature was then increased to 90 °C and stirred for 5 h. TLC showed that the starting material had completely reacted. The solution was evaporated to dryness and passed through a column chromatography (DCM:MeOH (v:v) = 100:1) to give 45 mg of white solid, with a yield of 40.9%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.04 (d, ¹H) J = 8.5 Hz, 1H), 7.99 (d, J = 1.7 Hz, 1H), 7.94 – 7.89 (m, 2H), 7.71 (s,1H), 7.50 (t, J = 7.8 Hz, 1H), 7.47 – 7.43 (m, 2H), 7.37 (t, J = 7.7 Hz, 1H), 7.22 (d, J = 7.8 Hz, 1H), 7.11 (d, J = 7.7 Hz, 1H), 6.94 (s, 1H), 5.00 (s, 2H), 4.52 (d, J= 6.0 Hz, 2H), 3.88 (s, 3H), 2.51 (s, 3H); MS m / z (ESI): 498.1 [M+H] + .

[0218] Example 23 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3-trifluoromethylbenzyl)acetamide, and the structural formula is:

[0219] The reaction formula and specific preparation steps for this compound are as follows:

[0220] Under argon protection, 2 g (11.42 mmol) of 3-trifluoromethylbenzylamine, 10 mL of DCM, and 1.62 mL (11.65 mmol) of triethylamine were added to a 100 mL single-necked flask. Then, 1.01 mL (11.65 mmol) of bromoacetyl bromide was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 2.5 g of a brown solid, which was used directly in the next reaction step.

[0221] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (98 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h. TLC showed that a large amount of the starting material was still unreacted. The temperature was then increased to 90 °C and stirred for 5 h. TLC showed that the starting material had completely reacted. The solution was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 30 mg of white solid, with a yield of 26.6%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.5 Hz, 1H), 7.97 (s, 1H), 7.70 (s, 1H), 7.53 – 7.47 (m, 3H), 7.43(dd, J = 14.6, 7.7 Hz, 3H), 7.22 (d, J = 7.8 Hz, 1H), 7.11 (d, J = 7.7 Hz, 1H),7.03 (s, 1H), 5.00 (s, 2H), 4.53 (d,J = 6.0 Hz, 2H), 2.51 (s, 3H); MS m / z(ESI):508.2[M+H] + .

[0222] Example 24 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(4-tert-butylbenzyl)acetamide, and the structural formula is:

[0223] The reaction formula and specific preparation steps for this compound are as follows:

[0224] Under argon protection, 2 g (12.25 mmol) of 4-tert-butylbenzylamine, 10 mL of DCM, and 1.74 mL (12.5 mmol) of triethylamine were added to a 100 mL single-necked flask. Then, 1.09 mL (12.5 mmol) of bromoacetyl bromide was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 2.5 g of a brown oily substance, which was used directly in the next reaction.

[0225] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (95 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 90 °C for 6 h, and TLC showed that the starting material had completely reacted. The solution was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 35 mg of a light-colored solid, with a yield of 31.8%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.97 (s, ¹H), 8.05 (d, ¹H) J = 8.5 Hz, 1H), 7.99 (d, J = 1.7 Hz, 1H),7.70 (s, 1H), 7.50 (t, J = 7.8 Hz, 1H), 7.46 (dd, J = 8.4, 1.7 Hz, 1H), 7.31 (d, J = 8.2 Hz, 2H), 7.23 (d, J = 7.7 Hz, 1H), 7.17 (d, J= 8.0 Hz, 2H), 7.10 (d, J = 7.7Hz, 1H), 4.97 (s, 2H), 4.45 (d, J = 5.8 Hz, 2H), 2.51 (s, 3H), 1.28 (s, 9H); MSm / z (ESI): 496.2[M+H] + .

[0226] Example 25 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(4-methoxybenzyl)acetamide, and the structural formula is:

[0227] The reaction formula and specific preparation steps for this compound are as follows:

[0228] Under argon protection, 2 g of 4-methoxybenzylamine (14.58 mmol), 10 mL of DCM, and 2.07 mL of triethylamine (14.87 mmol) were added to a 100 mL single-necked flask. Then, 1.3 mL of bromoacetyl bromide (14.87 mmol) was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 2.8 g of a brown solid, which was used directly in the next reaction.

[0229] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (86 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 90 °C for 6 h, and TLC showed that the starting material had completely reacted. The solution was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 16 mg of a light-colored solid, with a yield of 15.3%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.97 (s, ¹H), 8.04 (d, ¹H) J = 8.5 Hz, 1H), 7.97 (d, J = 1.7 Hz, 1H),7.69 (s, 1H), 7.50 (t, J = 7.7 Hz, 1H), 7.45 (dd, J = 8.4, 1.7 Hz, 1H), 7.20 (d, J= 7.8 Hz, 1H), 7.17 (d, J = 8.4 Hz, 2H), 7.10 (d, J = 7.7 Hz, 1H), 6.82 (d, J = 8.4Hz, 2H), 6.76 (s, 1H), 4.96 (s, 2H), 4.41 (d, J = 5.8 Hz, 2H), 3.77 (s, 3H), 2.51 (s, 3H); MS m / z (ESI): 470.2[M+H] + .

[0230] Example 26 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(4-fluorobenzyl)acetamide, and the structural formula is:

[0231] The reaction formula and specific preparation steps for this compound are as follows:

[0232] Under argon protection, 2 g (16 mmol) of 4-fluorobenzylamine, 10 mL of DCM, and 2.07 mL (16.3 mmol) of triethylamine were added to a 100 mL single-necked flask. Then, 1.3 mL (16.3 mmol) of bromoacetyl bromide was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 3.8 g of a brown solid, which was used directly in the next reaction step.

[0233] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (86 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 90 °C for 6 h, and TLC showed that the starting material had completely reacted. The solution was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 36 mg of a light-colored solid, with a yield of 35.4%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.4 Hz, 1H), 7.97 (d, J = 1.7 Hz, 1H),7.69 (s, 1H), 7.51 (t, J= 7.7 Hz, 1H), 7.45 (dd, J = 8.5, 1.7 Hz, 1H), 7.22 (dd, J = 8.3, 5.2 Hz, 3H), 7.12 (d, J = 7.7 Hz, 1H), 6.97 (t, J = 8.6 Hz, 2H), 6.89 (s,1H), 4.98 (s, 2H), 4.43 (d, J = 5.9 Hz, 2H), 2.52 (s, 3H); MS m / z (ESI): 458.2[M+H] + .

[0234] Example 27 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,4,6-trifluorobenzyl)acetamide, and the structural formula is:

[0235] The reaction formula and specific preparation steps for this compound are as follows:

[0236] Under argon protection, 2,4,6-trifluorobenzylamine (1 g, 6.21 mmol), 10 mL of DCM, and triethylamine (0.88 mL, 6.33 mmol) were added to a 100 mL single-necked flask. Bromoacetyl bromide (0.55 mL, 6.33 mmol) was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 1.3 g of a brown solid, which was used directly in the next reaction step.

[0237] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (94 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 90 °C for 6 h, and TLC showed that the starting material had completely reacted. The product was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 25 mg of a light-colored solid, with a yield of 22.8%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.4 Hz, 1H), 7.98 (d, J= 1.7 Hz, 1H),7.66 (s, 1H), 7.53 (t, J = 7.7 Hz, 1H), 7.46 (dd, J = 8.5, 1.6 Hz, 1H), 7.25 (s,1H), 7.12 (d, J = 7.6 Hz, 1H), 6.88 (s, 1H), 6.63 (t, J = 8.1 Hz, 2H), 4.92 (s,2H), 4.52 (d, J = 5.6 Hz, 2H), 2.52 (s, 3H); MS m / z (ESI): 494.2[M+H] + .

[0238] Example 28 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,3,6-trifluorobenzyl)acetamide, and the structural formula is:

[0239] The reaction formula and specific preparation steps for this compound are as follows:

[0240] Under argon protection, 2,3,6-trifluorobenzylamine (1 g, 6.21 mmol), 10 mL of DCM, and triethylamine (0.88 mL, 6.33 mmol) were added to a 100 mL single-necked flask. Bromoacetyl bromide (0.55 mL, 6.33 mmol) was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 1.4 g of a brown oily substance, which was used directly in the next reaction.

[0241] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (94 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 90 °C for 6 h, and TLC showed that the starting material had completely reacted. The solution was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 10 mg of a light-colored solid, with a yield of 9.1%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J= 8.4 Hz, 1H), 7.99 (d, J = 1.7 Hz, 1H),7.66 (s, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.46 (dd, J = 8.5, 1.7 Hz, 1H), 7.25 (s,1H), 7.11 (d, J = 7.7 Hz, 1H), 7.06 (tt, J = 9.1, 4.6 Hz, 1H), 6.94 (s, 1H), 6.84– 6.77 (m, 1H), 4.93 (s, 2H), 4.59 (d, J = 5.9 Hz, 2H), 2.51 (s, 3H); MS m / z(ESI):494.1[M+H] + .

[0242] Example 29 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3,5-difluoropyridin-4-methylene)acetamide, and the structural formula is:

[0243] The reaction formula and specific preparation steps for this compound are as follows:

[0244] Under argon protection, (3,5-difluoropyridin-4-yl)methylamine (1 g, 6.94 mmol), 10 mL of DCM, and triethylamine (0.98 mL, 7.08 mmol) were added to a 100 mL single-necked flask. Bromoacetyl bromide (0.62 mL, 7.08 mmol) was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 1 g of a brown solid, which was used directly in the next reaction step.

[0245] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (88 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h, and TLC showed that the starting material had completely reacted. The product was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 25 mg of a light-colored solid, with a yield of 23.6%. 1¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, 1H), 8.30 (s, 2H), 8.06 (d, 1H) J = 8.4 Hz, 1H), 7.97 (d, J =1.7 Hz, 1H), 7.67 (s, 1H), 7.54 (t, J = 7.7 Hz, 1H), 7.45 (dd, J = 8.5, 1.7 Hz, 1H), 7.24 (d, J = 8.0 Hz, 1H), 7.20 – 7.15 (m, 1H), 7.13 (d, J = 7.7 Hz, 1H), 4.94 (s, 2H), 4.61 (d, J = 6.0 Hz, 2H), 2.53 (s, 3H); MS m / z (ESI): 477.1[M+H] + .

[0246] Example 30 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(4-chlorobenzyl)acetamide, and the structural formula is:

[0247] The reaction formula and specific preparation steps for this compound are as follows:

[0248] Under argon protection, 2 g of 4-chlorobenzylamine (14.12 mmol), 10 mL of DCM, and 2.00 mL of triethylamine (14.41 mmol) were added to a 100 mL single-necked flask. Then, 1.3 mL of bromoacetyl bromide (14.41 mmol) was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 3.2 g of a light-colored solid, which was used directly in the next reaction step.

[0249] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (86 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 90 °C for 6 h, and TLC showed that the starting material had completely reacted. The solution was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 45 mg of a white solid, with a yield of 42.7%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.5 Hz, 1H), 7.96 (d, J = 1.7 Hz, 1H),7.69 (s, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.44 (dd, J = 8.5, 1.7 Hz, 1H), 7.29 (d, J = 8.1 Hz, 1H), 7.24 (s, 1H), 7.21 (d, J = 4.1 Hz, 1H), 7.19 – 7.16 (m, 2H), 7.12 (d, J = 7.7 Hz, 1H), 6.97 (d, J = 6.8 Hz, 1H), 4.97 (s, 2H), 4.46 – 4.43 (m,2H), 2.52 (s, 3H); MS m / z (ESI): 474.2[M+H] + .

[0250] Example 31 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(4-bromobenzyl)acetamide, and the structural formula is:

[0251] The reaction formula and specific preparation steps for this compound are as follows:

[0252] Under argon protection, 2 g (10.75 mmol) of 4-bromobenzylamine, 10 mL of DCM, and 2.00 mL (10.96 mmol) of triethylamine were added to a 100 mL single-necked flask. Then, 1.3 mL (10.96 mmol) of bromoacetyl bromide was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate solution, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 3.62 g of a white solid, which was used directly in the next reaction.

[0253] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (86 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 90 °C for 6 h, and TLC showed that the starting material had completely reacted. The solution was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 65 mg of a white solid, with a yield of 56.4%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.05 (d, ¹H) J = 8.5 Hz, 1H), 7.97 (d, J = 1.7 Hz, 1H),7.69 (s, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.46 – 7.45 (m, 1H), 7.44 – 7.43 (m,1H), 7.40 (d, J = 1.9 Hz, 1H), 7.20 (d, J = 7.8 Hz, 1H), 7.16 (d, J = 8.2 Hz, 1H),7.13 (s, 1H), 7.11 (s, 1H), 6.95 (s, 1H), 4.97 (s, 2H), 4.41 (d, J = 6.0 Hz, 2H), 2.52 (s, 3H); MS m / z (ESI): 520.1[M+H] + .

[0254] Example 32 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3-fluoropyridin-4-benzyl)acetamide, and the structural formula is:

[0255] The reaction formula and specific preparation steps for this compound are as follows:

[0256] Under argon protection, (3-fluoropyridin-4-yl)methylamine (0.5 g, 3.96 mmol), 10 mL of DCM, and triethylamine (0.56 mL, 4.04 mmol) were added to a 100 mL single-necked flask. Acetyl bromobromo (0.35 mL, 4.04 mmol) was added at -5 °C. The mixture was stirred at room temperature for 10 min, and TLC showed that the reaction was essentially complete. The mixture was extracted with ice water and DCM, washed with sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness at 5 °C to obtain 150 mg of a brown oil, which was used directly in the next reaction.

[0257] Under argon protection, intermediate 1 (65 mg, 0.222 mmol), the above-mentioned brominated product (83 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 90 °C for 6 h, and TLC showed that the starting material had completely reacted. The solution was evaporated to dryness and passed through a column chromatography column (DCM:MeOH (v:v) = 100:1) to give 5 mg of a white solid. MS m / z (ESI): 459.2 [M+H] + .

[0258] Example 33 This example provides a compound with the chemical name: 2-[4-(benzothiazol-6-yl)-3-(4-methylthiazol-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetamide, and the structural formula is:

[0259] The reaction formula and specific preparation steps for this compound are as follows:

[0260] Under argon protection, intermediate 2 (120 mg, 0.402 mmol), the bromide (160 mg, 0.603 mmol), potassium carbonate (112 mg, 0.804 mmol), and acetonitrile (10 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 4 h, and TLC showed that the starting material had completely reacted. Silica gel was added, and the mixture was directly evaporated to dryness. Column chromatography was performed with DCM / MeOH (v:v) = 200 / 1 as the eluent, which could separate the above isomers, giving a white solid. Hexane / EA = 4 / 1 (5 mL of mixed solvent) was added, the mixture was slurried, filtered, and 82 mg of white solid was obtained, yield: 43%. 1¹H NMR (500 MHz, deuterated chloroform) δ 9.01 (s, ¹H), 8.22 (s, ¹H), 8.12 (d, ¹H) J = 8.5Hz, 1H), 7.66 (s, 1H), 7.65 – 7.60 (m, 1H), 7.24 (t, J = 7.7 Hz, 1H), 6.91 –6.83 (m, 3H), 6.77 (s, 1H), 4.91 (s, 2H), 4.59 (d, J = 5.8 Hz, 2H), 2.43 (s,3H); MS m / z (ESI): 482.1[M+H] + .

[0261] Example 34 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(4-methylthiazolyl-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetamide

[0262] The reaction formula and specific preparation steps for this compound are as follows:

[0263] Under argon protection, intermediate 4 (120 mg, 0.409 mmol), the bromide (160 mg, 0.614 mmol), potassium carbonate (113 mg, 0.818 mmol), and acetonitrile (10 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 4 h, and TLC showed that the starting material had completely reacted. Silica gel was added, and the mixture was directly evaporated to dryness. Column chromatography was performed with DCM / MeOH (v:v) = 80 / 1 as the eluent, which could separate the above isomers, yielding a white solid. Hexane / EA = 4 / 1 (5 mL of mixed solvent) was added, the mixture was slurried, filtered, and washed with hexane / EA = 4 / 1 (5 mL) to give 80 mg of white solid, with a yield of 42%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.94 (dd, J = 4.2, 1.6 Hz, 1H), 8.43 – 8.33 (m, 3H), 8.20 (s, 1H), 7.63 (dd, J = 8.5, 4.2 Hz, 1H), 7.23 (td, J =7.6, 6.7, 1.9 Hz, 1H), 6.95 (s, 1H), 6.91 (s, 1H), 6.86 (t,J = 7.7 Hz, 2H), 4.93 (s, 2H), 4.59 (d, J = 5.8 Hz, 2H), 2.49 (s, 3H); MS m / z (ESI): 477.1[M+H] + .

[0264] Example 35 This example provides a compound with the chemical name: 2-[4-(quinoxaloline-6-yl)-3-(4-methylthiazolyl-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetamide, and the structural formula is:

[0265] The reaction formula and specific preparation steps for this compound are as follows:

[0266] Under argon protection, intermediate 5 (73 mg, 0.249 mmol), the bromide (99 mg, 0.373 mmol), potassium carbonate (69 mg, 0.498 mmol), and acetonitrile (5.0 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 4 h, and TLC showed that the starting material had completely reacted. Silica gel was added, and the mixture was directly evaporated to dryness. Column chromatography was performed with DCM / MeOH (v:v) = 200 / 1, which effectively separated the isomers, yielding a white solid. Hexane / EA = 4 / 1 (5 mL mixed solvent) was added, the mixture was slurried, filtered, and 60 mg of white solid was obtained, yield: 50%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.88 – 8.81 (m, 2H), 8.28 (d, J = 2.0 Hz, 1H), 8.08 (d, J = 8.7 Hz, 1H), 7.98 (dd, J = 8.7, 2.0 Hz, 1H), 7.76 (s, 1H), 7.26 –7.18 (m, 1H), 6.92 – 6.84 (m, 3H), 6.80 (s, 1H), 4.93 (s, 2H), 4.60 (d, J = 5.8Hz, 2H), 2.42 (d, J = 1.0 Hz, 3H); MS m / z (ESI):477.2[M+H] + .

[0267] Example 36 This example provides a compound with the chemical name: 2-[4-(1,4-benzoxan-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetamide, and the structural formula is:

[0268] The reaction formula and specific preparation steps for this compound are as follows:

[0269] Under argon protection, intermediate 6 (80 mg, 0.273 mmol), the bromide (108 mg, 0.409 mmol), potassium carbonate (75 mg, 0.546 mmol), and acetonitrile (10 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 2.5 h, and the reaction was visualized by TLC. Silica gel was added, the mixture was directly evaporated to dryness, and column chromatography was performed. The eluent, DCM / MeOH (v:v) = 100 / 1, was sufficient to separate the isomers, yielding a white solid. Hexane / EA = 4 / 1 (5 mL mixed solvent) was added, the mixture was slurried, centrifuged, and 60 mg of white solid was obtained, with a yield of 46%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 7.54 – 7.45 (m, 2H), 7.24 – 7.16 (m, 2H), 7.09 (d, J = 7.7 Hz, 1H), 6.88 – 6.82 (m,3H), 6.81 – 6.73 (m,3H), 4.89 (s, 2H), 4.56(d, J = 5.8 Hz, 2H), 4.29 – 4.20 (m, 4H), 2.58 (s, 3H); MS m / z (ESI): 477.1[M+H] + .

[0270] Example 37 This example provides a compound with the chemical name: 2-[4-(2,1,3-benzothiadiazol-5-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetamide, and the structural formula is:

[0271] The reaction formula and specific preparation steps for this compound are as follows:

[0272] Under argon protection, intermediate 7 (70 mg, 0.238 mmol), the bromide (95 mg, 0.358 mmol), potassium carbonate (66 mg, 0.476 mmol), and acetonitrile (5.0 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 2 h, and TLC showed that the starting material had completely reacted. Silica gel was added, and the mixture was directly evaporated to dryness. Column chromatography was performed with DCM / MeOH (v:v) = 100 / 1 as the eluent, which could separate the above isomers, yielding a white solid. Hexane / EA = 4 / 1 (5 mL of mixed solvent) was added, the mixture was slurried, filtered, and 50 mg of white solid was obtained, with a yield of 45%. 1 HNMR (500 MHz, deuterated chloroform) δ 7.98 (d, J = 1.6 Hz, 1H), 7.87 (d, J = 9.0 Hz, 1H),7.74 (s, 1H), 7.57 (dd, J = 8.6, 6.9 Hz, 2H), 7.39 (d, J = 7.7 Hz, 1H), 7.25 –7.18 (m, 1H), 7.12 (d, J = 7.7 Hz, 1H), 6.93 – 6.82 (m, 3H), 4.94 (s, 2H), 4.59(d, J = 5.8 Hz, 2H), 2.46 (s, 3H); MS m / z (ESI): 477.2[M+H] + .

[0273] Example 38 This example provides a compound with the chemical name: 2-[4-([1,2,4]triazolo[1,5-A]pyridin-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetamide, and the structural formula is:

[0274] The reaction formula and specific preparation steps for this compound are as follows:

[0275] Under argon protection, intermediate 8 (70 mg, 0.253 mmol), the bromide (95 mg, 0.38 mmol), potassium carbonate (66 mg, 0.506 mmol), and acetonitrile (5.0 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 4 h, and TLC showed that the starting material had completely reacted. Silica gel was added, and the mixture was directly evaporated to dryness. Column chromatography was performed with DCM / MeOH (v:v) = 30 / 1, which effectively separated the isomers, yielding a white solid. Hexane / EA = 4 / 1 (5 mL mixed solvent) was added, the mixture was slurried, filtered, and 50 mg of white solid was obtained, yield: 45%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.87 (s, 1H), 8.33 (s, 1H), 7.68 – 7.66 (m, 2H), 7.63 – 7.54 (m, 2H), 7.51 (d, J = 7.8 Hz, 1H), 7.27 – 7.23 (m, 1H), 7.11(d, J = 7.6 Hz, 1H), 6.91 – 6.85 (m, 3H), 4.91 (s, 2H), 4.59 (d, J = 5.9 Hz, 2H),2.45 (s, 3H); MS m / z (ESI):460.2[M+H] + .

[0276] Example 39 This example provides a compound with the chemical name: 2-[4-(thiophene[3,2-C]pyridin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetamide, and the structural formula is:

[0277] The reaction formula and specific preparation steps for this compound are as follows:

[0278] Under argon protection, intermediate 9 (70 mg, 0.239 mmol), the bromide (95 mg, 0.359 mmol), potassium carbonate (66 mg, 0.479 mmol), and acetonitrile (5.0 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 4 h, and TLC showed that the starting material had completely reacted. Silica gel was added, and the mixture was directly evaporated to dryness. Column chromatography was performed with DCM / MeOH (v:v) = 40 / 1, which effectively separated the isomers, yielding a white solid. Hexane / EA = 4 / 1 (5 mL of mixed solvent) was added, and the mixture was slurried and centrifuged to obtain 40 mg of a yellow solid, with a yield of 35%.1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.97 (s, ¹H), 8.39 (d, ¹H) J = 5.5 Hz, 1H), 7.81 (s,1H), 7.70 (d, J = 5.5 Hz, 1H), 7.63 (t, J = 7.7 Hz, 1H), 7.59 (s, 1H), 7.47 (d, J =7.7 Hz, 1H), 7.25 – 7.20 (m, 1H), 7.18 (d, J = 7.7 Hz, 1H), 6.89 – 6.79 (m,3H), 4.90 (s, 2H), 4.58 (d, J = 5.8 Hz, 2H), 2.62 (s, 3H); MS m / z (ESI): 476.2[M+H] + .

[0279] Example 40 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(pyridin-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetamide, and the structural formula is:

[0280] The reaction formula and specific preparation steps for this compound are as follows:

[0281] Under argon protection, intermediate 10 (100 mg, 0.359 mmol), bromide (123 mg, 0.467 mmol), potassium carbonate (100 mg, 0.718 mmol), and acetonitrile (5.0 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 4 h, and TLC showed that the starting material had completely reacted. Silica gel was added, and the mixture was directly evaporated to dryness. Column chromatography was performed with DCM / MeOH (v:v) = 50 / 1 as the eluent, yielding a white solid. Hexane / EA = 4 / 1 (5 mL of mixed solvent) was added, the mixture was stirred, filtered, and 80 mg of white solid was obtained, yield: 50%. 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.98 (s, ¹H), 8.60 (d, ¹H) J = 4.9 Hz, 1H), 8.06 (d, J = 8.5 Hz, 1H), 7.94(d, J= 1.8 Hz, 1H), 7.66 (s, 1H), 7.64 (td, J = 7.7, 1.8 Hz, 1H), 7.48 (d, J = 7.9Hz, 1H), 7.45 (dd, J = 8.4, 1.8 Hz, 1H), 7.26 – 7.18 (m, 2H), 6.89 – 6.90 (m,3H), 4.93 (s, 2H), 4.59 (d, J = 5.9 Hz, 2H); MS m / z (ESI): 462.2[M+H] + .

[0282] Example 41 This example provides a compound with the chemical name: 2-[4-(thiophene[3,2-C]pyridin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3,5-difluoropyridin-4-methylene)acetamide, and the structural formula is:

[0283] The reaction formula and specific preparation steps for this compound are as follows:

[0284] Under argon protection, intermediate 9 (65 mg, 0.222 mmol), the brominated product from Example 29 (88 mg, 0.333 mmol), potassium carbonate (62 mg, 0.444 mmol), and acetonitrile (5 mL) were added to a 100 mL single-necked flask. The mixture was refluxed at 85 °C for 6 h, and TLC showed complete reaction of the starting material. The solution was evaporated to dryness and passed through a column (DCM:MeOH (v:v) = 100:1) to give 33 mg of a light-colored solid, yield 31%. MS m / z (ESI): 477.2 [M+H] + .

[0285] Example 42 This example provides a compound with the chemical name: 2-[4-([1,2,4]triazolo[1,5-A]pyridin-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3,5-difluoropyridin-4-methylene)acetamide, and the structural formula is:

[0286] The reaction formula and specific preparation steps for this compound are as follows:

[0287] Under argon protection, intermediate 8 (70 mg, 0.253 mmol), the brominated product from Example 29 (100 mg, 0.38 mmol), potassium carbonate (66 mg, 0.506 mmol), and acetonitrile (5.0 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 4 h, and TLC showed that the starting material had completely reacted. Silica gel was added, and the mixture was directly evaporated to dryness. Column chromatography was performed with DCM / MeOH (v:v) = 30 / 1, which effectively separated the isomers, yielding a white solid. Hexane / EA = 4 / 1 (5 mL of mixed solvent) was added, the mixture was slurried, filtered, and 45 mg of white solid was obtained, yield: 38.8%.

[0288] 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.86 (s, 1H), 8.34 (s, 1H), 8.32 (s, 2H), 7.70 (s, 1H), 7.69 (d, J = 6.5 Hz, 1H), 7.62 (t, J = 7.7 Hz, 1H), 7.56 (d, J = 9.2 Hz, 1H), 7.51 (d, J = 7.8 Hz, 1H), 7.14 (d, J = 7.6 Hz, 1H), 7.10 (br, 1H), 4.93 (s, 2H), 4.62 (d, J = 6.0 Hz, 2H), 2.46 (s, 3H); MS m / z (ESI): 461.2 [M+H] + .

[0289] Example 43 This example provides a compound with the chemical name: 2-[4-(2,1,3-benzothiadiazol-5-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3,5-difluoropyridin-4-methylene)acetamide, and the structural formula is:

[0290] The reaction formula and specific preparation steps for this compound are as follows:

[0291] Under argon protection, intermediate 7 (70 mg, 0.238 mmol), the brominated product from Example 29 (95 mg, 0.358 mmol), potassium carbonate (66 mg, 0.476 mmol), and acetonitrile (5.0 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 2 h, and TLC showed complete reaction of the starting material. Silica gel was added, and the mixture was directly evaporated to dryness. Column chromatography was performed with DCM / MeOH (v:v) = 100 / 1 as the eluent, which could separate the above isomers, yielding a white solid. Hexane / EA = 4 / 1 (5 mL mixed solvent) was added, the mixture was slurried, filtered, and 50 mg of white solid was obtained, with a yield of 45%. MS m / z (ESI): 478.2 [M+H] + .

[0292] Example 44 This example provides a compound with the chemical name: 2-[4-(benzothiazo-6-yl)-3-(4-methylthiazo-2-yl)-1H-pyrazole]-N-(3,5-difluoropyridine-4-methylene)acetamide, and the structural formula is:

[0293] The reaction formula and specific preparation steps for this compound are as follows:

[0294] Under argon protection, intermediate 2 (120 mg, 0.402 mmol), the brominated product from Example 29 (160 mg, 0.603 mmol), potassium carbonate (112 mg, 0.804 mmol), and acetonitrile (10 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 4 h, and TLC showed complete reaction of the starting material. Silica gel was added, and the mixture was directly evaporated to dryness. Column chromatography was performed with DCM / MeOH (v:v) = 200 / 1, which effectively separated the isomers, yielding a white solid. Hexane / EA = 4 / 1 (5 mL mixed solvent) was added, and the mixture was slurried, filtered, and 43 mg of white solid was obtained, yield: 38%. MS m / z (ESI): 483.1 [M+H] + .

[0295] Example 45 This example provides a compound with the chemical name: 2-[4-(naphthidin-2-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(3,5-difluoropyridin-4-methylene)acetamide, and the structural formula is:

[0296] The reaction formula and specific preparation steps for this compound are as follows:

[0297] Under argon protection, intermediate 3 (50 mg, 0.174 mmol), bromoacetamide raw material from Example 29 (69 mg, 0.261 mmol), potassium carbonate (48 mg, 0.348 mmol), and acetonitrile (3 mL) were added to a 25 mL single-necked flask. The mixture was refluxed at 80 °C for 6 h. TLC showed that the raw material had reacted completely, and two new spots were formed. Ice water and DCM were added and separated. The mixture was evaporated to dryness and then separated by Flash chromatography (DCM:MeOH (v:v) = 30:1) to obtain a light-colored solid. 1 mL of n-heptane was added, and the mixture was evaporated to dryness to obtain 30 mg of a light yellow solid, with a yield of 37%.

[0298] 1 ¹H NMR (500 MHz, deuterated chloroform) δ 8.93 (d, J = 3.9 Hz, 1H), 8.35 – 8.28 (m, 3H), 8.23 ​​(d, J = 8.8 Hz, 1H), 8.14 (s, 1H), 7.77 (d, J = 8.8 Hz, 1H), 7.67 – 7.60 (m, 2H), 7.49 (d, J = 7.8 Hz, 1H), 7.19 (t, J = 7.9 Hz, 2H), 4.96 (s, 2H), 4.61 (d, J = 6.0 Hz, 2H), 2.51 (s, 3H); MS m / z (ESI): 472.2 [M+H] + .

[0299] Example 46 This example provides a compound with the chemical name: 2-[4-(benzo[d]thiazo-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazol-1-yl]-N-(2,6-difluorobenzyl)acetylthioamide, and the structural formula is:

[0300] The reaction formula and specific preparation steps for this compound are as follows:

[0301] Under nitrogen protection, the compound from Example 9 (20 mg, 0.042 mmol) was added to a 5 mL solution of Lawson's reagent (21 mg, 0.053 mmol) in toluene. After heating under reflux for 6 hours, the solvent was removed under reduced pressure, and the residue was purified by column chromatography to give a white solid (8 mg, 38.8%). MS m / z (ESI): 492.2 [M+H] + .

[0302] Example 47 This example provides a compound with the chemical name: 2-[4-([1,2,4]triazolo[1,5-A]pyridin-6-yl)-3-(6-methylpyridin-2-yl)-1H-pyrazole]-N-(2,6-difluorobenzyl)acetylthioamide, and its structural formula is:

[0303] The reaction formula and specific preparation steps for this compound are as follows:

[0304] Under nitrogen protection, the compound (20 mg, 0.044 mmol) prepared in Example 38 was added to a 5 mL solution of Lawson's reagent (21 mg, 0.053 mmol) in toluene. After heating under reflux for 6 hours, the solvent was removed under reduced pressure, and the residue was purified by column chromatography to give a white solid (6 mg, 28.6%). MS m / z (ESI): 476.2 [M+H] + .

[0305] Example 48 This example provides a compound with the chemical name: N-[(3,5-difluoropyridin-4-yl)methyl]-2-[3-(6-methylpyridin-2-yl)-4-(quinolin-4-yl)-1H-pyrazol-1-yl]acetamide, and its structural formula is:

[0306] The reaction formula and specific preparation steps for this compound are as follows:

[0307] Under argon protection, 4-[3-(6-methyl-2-pyridyl)-1H-pyrazol-4-yl]quinoline (50 mg, 0.175 mmol), bromoacetamide from Example 29 (102 mg, 0.385 mmol), potassium carbonate (68 mg, 0.49 mmol), and acetonitrile (10 mL) were added to a 50 mL single-necked flask. The mixture was refluxed at 85 °C for 5 h, and TLC showed that the reaction was complete. Silica gel was added, the mixture was directly evaporated to dryness, and column chromatography was performed. The eluent was DCM / MeOH = 40 / 1 (volume ratio) to obtain a white solid product. Hexane / EA = 4 / 1 (5 mL mixed solvent) was added, the mixture was slurried, centrifuged, and 28 mg of white solid was obtained.

[0308] 1¹H NMR (500 MHz, deuterated chloroform) δ 8.88 (d, J = 4.4 Hz, 1H), 8.32 (s, 2H), 8.14 (d, J = 8.4 Hz, 1H), 7.72 – 7.65 (m, 3H), 7.37 (td, J = 7.8, 2.9 Hz, 2H), 7.30 (d, J = 4.4 Hz, 1H), 7.09 (d, J = 7.8 Hz, 1H), 6.97 (d, J = 7.6 Hz, 1H), 5.01 (s, 2H), 4.65 (d, J = 6.0 Hz, 2H), 2.25 (s, 3H); MS m / z (ESI): 471.2 [M+H] + .

[0309] Biological evaluation test: Test Example 1: Inhibitory effect of the compound of the present invention on TGFβRI / ALK5 kinase activity. This invention employs the following experimental methods to determine the inhibitory effects of the compounds in the examples and AGMB-129 on TGFβRI / ALK5 kinase activity. AGMB-129 is an ALK5 inhibitor developed by Agomab Therapeutics NV, and its chemical structure is [not specified]. ; Experimental Methods: Compounds were serially diluted with DMSO in 384 PP compound dilution plates (Beckman, catalog number: 001-14555). 0.1 μL of the compound was transferred to 384 reaction microplates (Revity, catalog number: 6007290) using an Echo 655 microplate, ensuring a final DMSO concentration of 1% (double replicate). Proteins and ATP / substrate were prepared using 1× kinase reaction buffer (50 mM Hepes, 10 mM MgCl2, 0.01% Brij35, 1 mM EGTA, 2 mM DTT). Transfer 5 μL of ALK5 (5 nM, manufacturer: ICE, catalog number: S2405T-H57G) solution to a 384 assay plate, centrifuge at 1000 rpm for 60 seconds using a microplate centrifuge (model: Platepro3200, manufacturer: Monad), and incubate at 25°C for 10 minutes in a biochemical incubator (model: LRH-250F, manufacturer: Bluepard). Prepare a 2× substrate (TGFβR tide: 0.1 mg / mL) and ATP (1 μM, manufacturer: Promega, catalog number: V915B) mixture using kinase reaction buffer. Add 5 μL of the substrate and ATP mixture to the reaction plate and start the reaction, centrifuging at 1000 rpm for 1 minute. Incubate at 25°C for 180 minutes. Transfer 5 μL of ADP-GloReagent to a 384 assay plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes. Transfer 10 μL of the test solution to a 384 test plate, centrifuge at 1000 rpm for 1 minute, incubate at 25°C for 40 minutes, and read the chemiluminescence signal using a multi-functional microplate reader (model: PHERAstar FSX, manufacturer: BMG). The negative control (containing 1% DMSO, enzyme, substrate, and ATP) was set to 0% inhibition rate, and the positive control (containing 1% DMSO, 1× kinase buffer, substrate, and ATP) was set to 100% inhibition rate. The inhibition rate of each test solution was calculated, and the data was analyzed using software. The IC50 of the compound was obtained using a non-linear fitting formula. 50 (Half-maximal inhibitory concentration). The IC50 of the compound was determined using the method described above. 50 The values ​​are shown in Table 1 below.

[0310] Table 1. IC50 of the compounds of this invention on the inhibition of ALK5 kinase activity 50 value

[0311] As shown in Table 1, the compounds of the present invention have a significant inhibitory effect on ALK5 kinase activity, IC50... 50 Reaching nanomolar levels, it exhibits superior inhibitory effects compared to AGMB-129, with an IC50 of [missing value] for inhibiting ALK5 kinase activity.50 The value is 0.879-5.23 nM.

[0312] Test Example 2: Bidirectional Permeability Experiment of Caco-2 Cell Monolayer The bidirectional permeability of the compounds in some embodiments of this invention to Caco-2 cell monolayers was tested using the following method: (1) Reference compounds, the names of which are shown in Table 2 below: Table 2. Names of Reference Compounds

[0313] (2) Cell information Caco-2 cells purchased from ATCC at 1.00 × 10⁻⁶ 5 cells / cm 2 Cells were seeded at a density of [insert density here] onto polyethylene (PET) membranes in 96-well Corning insert plates, and the culture medium was changed every 4-5 days until a confluent cell monolayer was formed on days 21-28. Specific experimental parameters are shown in Table 3 below.

[0314] Table 3 Experimental Instrument Parameters

[0315] (3) Transport buffer This invention uses HBSS-Hank's balanced salt solution at pH 7.40 and 10.0 mM HEPES (2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid) as the transport buffer.

[0316] (4) Experimental methods Test compounds were tested in a bidirectional repeat at a concentration of 2.00 μM. Digoxin was tested in a bidirectional repeat at a concentration of 10.0 μM, while naldolol and metoprolol were tested in a repeat from A to B at a concentration of 2.00 μM. The final DMSO concentration was adjusted to below 1.0%. The reaction plate was incubated in a CO2 incubator at 37.0 °C and 5% CO2 saturated humidity for 2 hours without shaking. After mixing all samples with the stop solution, they were centrifuged at 3220 g for 10 minutes. The concentrations of test and control compounds in all samples were semi-quantitatively determined using LC-MS / MS, based on the peak area ratio of analyte / internal standard. After the transport experiment, the integrity of the Caco-2 cell monolayer was detected using a fluorescein yellow rejection assay.

[0317] (5) Data analysis methods Apparent permeability coefficient P app (cm / s) Calculated using the following formula: Papp = (dC r / dt) × V r / (Area × C0) Among them, dC r / dt represents the change in the cumulative concentration of the compound in the receiving chamber over time; V r The volume of solution in the receiving chamber is 0.0750 mL on the top side and 0.250 mL on the base side; Area is the transport surface area, i.e., the area of ​​the monolayer membrane is 0.0804 cm². 2 C0 represents the initial concentration in the donor chamber.

[0318] The formula for calculating the external discharge ratio is: Emission ratio (ER) = P app (BA) / P app (AB) AB refers to the direction from A to B, that is, from the apical side to the basal side, which mainly simulates intestinal absorption, that is, the absorption of substances from the intestinal lumen into the blood; BA refers to the direction from B to A, that is, from the basal side to the apical side, which mainly simulates the pumping of substances from the blood into the intestinal lumen.

[0319] The experimental data on the bidirectional permeability of Caco-2 cell monolayers obtained according to the above testing method are shown in Table 4 below.

[0320] Table 4. Experimental data on bidirectional permeability of Caco-2 cell monolayers.

[0321] In Table 4, ND indicates undetectable; P app (AB) < 0.500 (×10 -6 The value (cm / s) indicates a "low-permeability" compound, 0.500≤P app (AB) < 4.00 (×10 -6 (cm / s) indicates a "medium osmosis" compound, P app (AB) ≥ 4.00 (×10 - 6 An ER of 1 cm / s indicates a "high-permeability" compound; an ER ≥ 2.00 indicates that the compound may be an efflux transporter substrate, while an ER < 2.00 indicates that the compound may not be an efflux transporter substrate.

[0322] ALK5 inhibitors have high systemic toxicity, and the less the amount of compound absorbed into the bloodstream via the intestines, the better. Therefore, the P of this series of compounds... app(AB) should be as low as possible, meaning less intestinal absorption is better, while the efflux ratio (ER) should be as high as possible, meaning more efflux is better. Table 4 shows that the bidirectional permeability experiment data of the compounds of this invention in Caco-2 cell monolayers shows that all the illustrated examples and AGMB-129 have P... app (AB) are both higher than 4.00 (×10) -6 The concentration of permeability (cm / s) indicates that these are highly permeable compounds, meaning that all related compounds are highly absorbable in the intestines. Examples 10, 13, 29, 38, 42, 45, and 48, as well as AGMB-129, have efflux ratios (ER) higher than 2.00, indicating that these compounds are highly effluxed. Compared to AGMB-129, regarding P... app Regarding (AB), Examples 10, 13, 29, 38, 42, 45, and 48 have relatively small values, meaning less absorption into the bloodstream and thus a lower risk of systemic exposure. In terms of efflux ratio (ER), Examples 10, 13, 29, 38, 42, 45, and 48 are greater than AGMB-129, indicating that the series of compounds exhibits higher efflux than AGMB-129, implying a lower risk of systemic exposure. Therefore, based on the bidirectional permeability test results of Caco-2 cell monolayers, the series of compounds, including Examples 10, 13, 29, 38, 42, 45, and 48, are superior to AGMB-129 and are more ideal organ-restricted ALK5 inhibitors with lower systemic exposure risk.

[0323] Test Example 3: Stability Test of Compound in Liver Microsomes 1) Information on the control compounds used in the test is shown in Table 5 below.

[0324] Table 5. Information on control compounds

[0325] The stock solutions of the test compound and the control compound were diluted to 100 μM with acetonitrile.

[0326] 2) Liver microsomal information: The liver microsomal information used in the test is shown in Table 6 below.

[0327] Table 6 Liver microsomal information

[0328] Liver microsomes were diluted to a concentration of 0.56 mg / mL using 100 mM potassium phosphate buffer (PB buffer).

[0329] 3) Auxiliary factor information: The auxiliary factor information used in the test is shown in Table 7 below.

[0330] Table 7. Auxiliary Factor Information

[0331] Weigh an appropriate amount of reduced nicotinamide adenine dinucleotide phosphate (NADPH) powder and dilute it with 10 mM MgCl2 solution to a concentration of 10 mM.

[0332] 4) Termination solution Cold (4°C) acetonitrile (ACN) contains 250 nM tolbutamide and 250 nM labetalol as internal standards (IS).

[0333] 5) Experimental methods 5.1 Preheat the empty “incubation” plates T60 and NCF60 at 37°C for 10 minutes.

[0334] 5.2 Transfer the liver microsomal working solution (445 μL) to preheated “Incubation” plates T60 and NCF60, and then incubate at 37°C for 10 minutes.

[0335] 5.3 Transfer the microparticle working solution (54 µL) to a Blank60 plate, and then add 6 µL of NADPH cofactor and 180 µL of stop solution to each well.

[0336] 5.4 Add the compound working solution (5 μL) to the “incubation” plates (T60 and NCF60) containing liver microsomes.

[0337] 5.5 For the “Incubation” plate NCF60, add 50 µL of PB buffer and incubate the plate at 37°C for 60 minutes.

[0338] 5.6 Add the stop solution (180 µL) and NADPH working solution (6 µL) to the T0 plate. Then, remove the mixture (54 µL) from the "Incubation" plate T60 and transfer it to the T0 plate.

[0339] 5.7 For the “Incubation” plate T60, add NADPH working solution (44 µL) and incubate at 37°C for 60 minutes.

[0340] 5.8 At 5, 15, 30, 45 and 60 minutes, transfer 60 μL of sample at each time point to a well containing 180 μL of stop solution and mix.

[0341] 5.9 All sampling plates were shaken for 10 minutes and then centrifuged at 3220 ×g for 20 minutes at 4°C.

[0342] 5.10 Transfer the supernatant (80 µL) to 240 µL of pure water and mix on a plate shaker for 10 minutes.

[0343] 5.11 Before performing LC-MS / MS analysis, each bioanalytical plate was sealed and shaken for 10 minutes.

[0344] 6) Data Analysis The remaining percentage of the compound after incubation is calculated using the following formula:

[0345] The liver microsomal stability data of the compounds obtained according to the above test methods are shown in Table 8 below.

[0346] Table 8. Hepatic microsomal stability data of the compounds

[0347] As shown in Table 8, the stability test data of the compounds in liver microsomes of the present invention showed that after 60 minutes of incubation in human and rat liver microsomes, the prototype compounds of all examples and AGMB-129 were essentially undetectable. This indicates that the examples and AGMB-129 shown in the present invention are very unstable in human and rat liver microsomes, meaning that even if a small portion of the compound enters the bloodstream, the liver can quickly clear it, implying a low risk of systemic exposure. In particular, no residual prototype compounds were detected in Examples 6, 10, 13, 29, 38, 48, and AGMB-129 after 60 minutes of incubation in human liver microsomes, suggesting that this series of examples has a low risk of systemic exposure and low toxicity.

[0348] The embodiments of the present invention have been described in detail above. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. The compound represented by Formula I, Or its pharmaceutically acceptable salts, esters, deuterated compounds, isotopically labeled derivatives, solvates, crystalline compounds, stereoisomers, and prodrugs; in, Cycle A is selected from 5-10 member heteroaryl groups; the heteroaryl group is optionally unsubstituted or substituted with one or more R groups. a replace; Ring B is selected from 8- to 15-membered heteroaryl groups, wherein ring B contains at least one heteroatom selected from N, S, and O; the heteroaryl group is optionally unsubstituted or substituted with one or more R groups. a replace; When the heteroatom in ring B is N and the number of heteroatoms is 1, Y is not CH; Each R1 is independently selected from -COOR c Halogen, cyano, carboxyl, sulfonium pentafluoride, C 1-8 Alkyl, C 1-8 Alkoxy or C 3-8 Cycloalkyl; the alkyl, alkoxy, or cycloalkyl group is optionally unsubstituted or substituted with one or more R groups. b replace; R2 is selected from hydrogen, halogen, or C. 1-8 Alkyl; the alkyl group is optionally unsubstituted or substituted with one or more R b replace; Each R a Each is independently selected from halogens, deuterium, and carbon. 1-8 Alkyl, C 3-8 cycloalkyl or C 1-8 Alkoxy; the alkyl, alkoxy, or cycloalkyl group is optionally unsubstituted or substituted with one or more R groups. b replace; R c C 1-8 Alkyl; the alkyl group is optionally unsubstituted or substituted with one or more R b replace; Each R b Each is independently selected from halogens or deuterium; X is selected from O or S; Y is selected from CH or N; n is an integer selected from 0 to 4.

2. The compound according to claim 1, characterized in that: Each R1 is independently selected from fluorine, chlorine, bromine, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoroethyl, cyano, carboxyl, methyl, deuterated methyl, ethyl, propyl, isopropyl, cyclopropyl, tert-butyl, monofluoromethoxy, difluoromethoxy, trifluoromethoxy, methoxy, deuterated methoxy, ethoxy, propoxy, or isopropoxy.

3. The compound according to claim 1, characterized in that: R2 is selected from C substituted with hydrogen, fluorine, chlorine, bromine, or halogen. 1-5 alkyl.

4. The compound according to claim 1, characterized in that: Each R a Each is independently selected from fluorine, chlorine, bromine, methyl, deuterated methyl, ethyl, propyl, isopropyl, cyclopropyl, monofluoromethoxy, difluoromethoxy, trifluoromethoxy, methoxy, deuterated methoxy, ethoxy, propoxy, or isopropoxy.

5. The compound according to claim 1, characterized in that: Ring A is selected from a thiazole ring or a pyridine ring; preferably, ring A is selected from... or R3 is selected from H, fluorine, chlorine, bromine, methyl, deuterated methyl, ethyl or propyl.

6. The compound according to claim 1, characterized in that: Ring B is selected from hexa-6 heteroaryl or penta-6 heteroaryl; preferably, ring B is selected from benzothiazole, naphthidine, 2,1,3-benzothiadiazole, [1,2,4]triazolo[1,5-A]pyridine, quinoxaline, benzopyridine, thiophene[3,2-C]pyridine or 1,4-benzodioxane; Preferably, ring B is selected from , , , , , , or .

7. The compound according to claim 1, characterized in that: The compounds represented by Formula I are selected from: 。 8. A method for preparing the compound represented by Formula I according to any one of claims 1 to 7, characterized in that: When X in the compound represented by Formula I is O, the preparation method includes the following steps: The compound of formula Ia is reacted with the compound of formula Ib to obtain the compound shown in formula I; And optionally, it also includes the following steps: The compound of formula I obtained is reacted with Lawson's reagent to give the compound of formula I with X as S; The structural formula of the compound of formula Ia is: ; The structural formula of the compound of formula Ib is: ; Wherein, Z is selected from Br or Cl; Ring A, ring B, R1, R2, X, Y, and n are as defined in any one of claims 1 to 7.

9. The method for preparing the compound represented by Formula I according to claim 8, characterized in that: The compound of formula Ib is obtained by making... It is prepared by reacting with bromoacetyl bromide or chloroacetyl chloride.

10. A pharmaceutical composition, characterized in that: Includes pharmaceutically acceptable excipients and compounds of Formula I as described in any one of claims 1 to 7, or their pharmaceutically acceptable salts, esters, deuterated compounds, isotopically labeled derivatives, solvates, crystalline compounds, stereoisomers, and prodrugs.

11. The pharmaceutical composition according to claim 10, characterized in that: The dosage form of the pharmaceutical composition is selected from tablets, lozenges, tablets, aqueous suspensions, oil suspensions, water-oil suspensions, powders, granules, emulsions, hard capsules, soft capsules, syrups, or elixirs. And / or, the pharmaceutically acceptable excipients include at least one of the following conventional in the pharmaceutical field: solvents, inert diluents, dispersants, granulating agents, surfactants, emulsifiers, disintegrants, binders, preservatives, buffers, lubricants, excipients, colorants, coating agents, sweeteners, flavoring agents, and aromatizers.

12. The use of the compound of Formula I according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, its ester, its deuterated compound, its isotopically labeled derivative, its solvate, its crystalline compound, its stereoisomer, its prodrug, or the pharmaceutical composition according to any one of claims 10 to 11, in the preparation of a medicament for the prevention or treatment of a disease improved by inhibiting transforming growth factor-β receptor I / ALK5.

13. The application according to claim 12, characterized in that: The diseases mentioned include gastrointestinal diseases, cancer, fibrotic skin diseases, fibrotic eye diseases, pulmonary fibrosis, or liver fibrosis.

14. The application according to claim 13, characterized in that: The gastrointestinal diseases mentioned are selected from Crohn's disease and ulcerative colitis; And / or, the cancer is selected from stomach cancer, esophageal cancer, and colorectal cancer; And / or, the fibrotic skin diseases include scleroderma, nephrotic fibrotic skin disease, mixed connective tissue disease, sclerosing myxedema, sclerosing disease, and eosinophilic fasciitis; And / or, the fibrotic eye disease is selected from dry eye disease, age-related macular degeneration, corneal and conjunctival scarring, postcataract fibrosis, proliferative vitreoretinopathy, or proliferative diabetic retinopathy.

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