Compounds that can be used to lower uric acid

Abstract

The present invention discloses a class of compounds that can be used to lower uric acid. These compounds are represented by general formula (I) or pharmaceutically acceptable salts thereof. They can significantly reduce serum uric acid levels in hyperuricemia rat models and have potential application value in anti-gout and anti-hyperuricemia drugs. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a class of compounds that can be used to lower uric acid. Background Art

[0002] Xanthine oxidase (XO) is an important target for drug treatment of hyperuricemia and gout. Hypoxanthine in the human body is metabolized into xanthine, which is further metabolized into uric acid. XO plays a crucial role in the formation of uric acid. Inhibiting XO activity inhibits the conversion of hypoxanthine, xanthine, and uric acid, thereby reducing the concentration of uric acid in human serum. Therefore, inhibiting XO activity is key to inhibiting uric acid production.

[0003] Generally speaking, a blood uric acid level exceeding 420 μmol / L in men and exceeding 360 μmol / L in women is called hyperuricemia. Currently, hyperuricemia has become the second largest metabolic disease after diabetes, seriously threatening human health. Hyperuricemia is not only an important biochemical basis for gout, but is also closely related to the occurrence of hypertension, hyperlipidemia, atherosclerosis, obesity, and insulin resistance. In the past few decades, due to the prevalence of comorbidities that can promote hyperuricemia (such as hypertension, obesity, metabolic syndrome, type 2 diabetes, and chronic kidney disease), the incidence of gout has also gradually increased (Khanna D, Fitzgerald JD, Khanna PP, et al. American College of Rheumatology Guidelines for Management of Gout. Part 1: Systematic Nonpharmacologic and Pharmacologic Therapeutic Approaches to Hyperuricemia [J]. Arthritis Care & Research, 2012, 64 (10): 1432-1446).

[0004] The increasing incidence of hyperuricemia and gout poses a serious threat to human health and warrants early intervention and treatment. However, the currently available uric acid-lowering medications are very limited, have significant side effects, and suffer from poor patient compliance. Currently, there are three main classes of drugs used to treat hyperuricemia and gout: urate anion transporter 1 (URAT1) inhibitors, xanthine oxidase inhibitors, and urate oxidase inhibitors.

[0005] URAT1 inhibitors primarily act on the urate transporter in the renal proximal tubule, inhibiting uric acid reabsorption and increasing its excretion, thereby reducing uric acid concentration in the body. This class of drugs includes benzbromarone, lesinurad, and probenecid. Benzbromarone is an effective uricosuric drug approved for marketing in many countries, but not in the United States. Due to severe hepatotoxicity, benzbromarone was withdrawn from several European countries in 2003. Due to racial disparities in liver-related adverse events associated with benzbromarone, the Chinese Guidelines for the Diagnosis and Treatment of Hyperuricemia and Gout (2019) recommend benzbromarone as a first-line uric acid-lowering drug. Lesinurad, approved for marketing in the United States in 2015, carries a black box warning regarding the risk of acute renal failure and cardiovascular disease (potentially fatal), and its efficacy is far inferior to that of benzbromarone, requiring combination therapy with allopurinol. Probenecid is the first choice of uricosuric drug in single-agent uric acid-lowering treatment in the US guidelines, but its application is limited due to multiple significant interactions with some commonly used drugs (such as nonsteroidal anti-inflammatory drugs, β-lactam drugs, heparin, etc.).

[0006] Xanthine oxidase inhibitors primarily include allopurinol and febuxostat. Allopurinol has been widely used clinically since its approval by the US FDA in 1966. Currently, it remains the first-line treatment for gout recommended in most national gout guidelines. However, allopurinol is poorly effective, inhibiting only the reduced form of XO, but not the oxidized form. Studies have shown that even at the maximum dose of allopurinol, the rate of subjects reaching the treatment endpoint is less than 50% (Robert M, Douglas CA, Scott B. Less than half of patients treated with high-dose allopurinol reach serum uric acid target[J]. ACR / ARHP Annual Meeting, 2017, Abstract Number: 1120). In addition, it can also cause rash and other rare but fatal side effects, including Stevens-Johnson syndrome and toxic epidermal necrolysis, with a mortality rate of approximately 10% to 30% (Bocquet H, Bagot M, Roujeau JC. Drug-induced pseudolymphoma and drug hypersensitivity syndrome (drug rash with eosinophilia and systemic symptoms: DRESS[J]. Seminars in Cutaneous Medicine and Surgery, 1996, 15(4): 250-257). Allopurinol has been reported to cause acute liver damage in clinical practice. Allopurinol should be used with caution in the treatment of patients with hyperuricemia and liver disease (Imai H, Kamei H, Onishi Y, et al. Successful living-donor liver transplantation for cholestatic liver failure induced by allopurinol: case report[J]. Transplantation Proceedings, 2015, 47(9): 2778-2781). Other side effects of allopurinol include stomach discomfort, nausea, abdominal pain, diarrhea, leukopenia and thrombocytopenia, headache, fever, loss of appetite, weight loss, painful urination, hematuria, itching, and drowsiness.

[0007] Febuxostat is a non-purine XO inhibitor developed by Teijin of Japan. It inhibits both the oxidized and reduced forms of XO and exhibits significantly higher activity than allopurinol. It was launched in Europe in 2008 and in the United States in 2009. As febuxostat's clinical use continues to expand, reports of cardiovascular adverse reactions in the treatment of hyperuricemia have increased. Due to its cardiovascular toxicity (such as the risk of sudden death), the US Food and Drug Administration required the addition of a black box warning to its drug label and adjusted its prescribing information in 2019, changing its designation from first-line to second-line. A study of 6,190 gout patients published in the New England Journal of Medicine in March 2018 found that after a mean of 32 months of treatment, the overall risk of adverse cardiovascular events was similar between the febuxostat and allopurinol groups (HR 1.03, 95% CI, 0.87-1.23). However, all-cause and cardiovascular mortality were higher in the febuxostat group than in the allopurinol group. Patients in the febuxostat group had a 34% increased risk of cardiovascular mortality (HR 1.34, 95% CI, 1.03-1.73) and a 22% increased risk of all-cause mortality (HR 1.22, 95% CI, 1.01-1.47). Sudden cardiac death was the most common cause of cardiovascular death, with 83 cases (2.7%) in the febuxostat group and 56 cases (1.8%) in the allopurinol group (William B, Kenneth G, Michael A, et al. Cardiovascular safety of febuxostat or allopurinol in patients with gout [J]. The New England Journal of Medicine, 2018, 378: 1200-1210). In addition, febuxostat may also cause serious gastrointestinal toxicity, renal toxicity, and liver function abnormalities.

[0008] Pegylated recombinant uricase is the leading urate oxidase drug currently on the market, administered via intravenous injection. The FDA has issued multiple black box warnings regarding the drug, which is associated with severe immune-allergic side effects in 20%-40% of patients. Its efficacy is modest, with only 47% of patients achieving a therapeutic endpoint of less than 0.36 mmol / L.

[0009] Over the past few decades, progress in drug development for hyperuricemia and gout has been slow. However, with the increasing incidence of these conditions, the development of therapeutics has attracted increasing attention from researchers. The design of new drugs targeting xanthine oxidase has also gained widespread attention, with several compounds entering clinical trials. However, these compounds still face significant toxic side effects, requiring further research. Therefore, highly effective and low-toxic XO inhibitors hold great potential for development and application. Summary of the Invention

[0010] The purpose of the present invention is to provide a compound having xanthine oxidase inhibitory activity based on the prior art.

[0011] Another object of the present invention is to provide the use of the above compound in the field of medicine.

[0012] The purpose of the present invention can be achieved by the following measures:

[0013] The compound represented by general formula (I) or a pharmaceutically acceptable salt thereof,

[0014]

[0015] in,

[0016] R is C 1-6 Alkyl, substituted C 1-6 Alkyl, C 3-6 Cycloalkyl, substituted C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl or substituted C 3-6 Heterocycloalkyl; wherein the substituents in each group involved in R are selected from deuterium, cyano, nitro, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or C 3-6 one or more of heterocycloalkyl;

[0017] Ar is a substituted or unsubstituted group: The substituents in the Ar group are selected from deuterium, hydroxyl, halogen, C 1-4 Alkyl or C 1-4 One or more of alkoxy groups;

[0018] Y is O or NR 3 ,

[0019] R 1 is a connecting bond or a substituted or unsubstituted C 1-6 Alkylene or substituted or unsubstituted C 2-12 Alkenylene, R 1 The substituents in the group are selected from deuterium, hydroxyl, amino, cyano, halogen, C 1-4 Alkyl or C 1-4 one or more of alkoxy groups;

[0020] R 2 is hydrogen, nitrooxy, carboxyl or substituted or unsubstituted following groups: dioxol-2-one, C 4-12 Condensed heteroaromatic ring group, C 4-16 Fused heteroaromatic ring pyrazolylcarbonyloxy, C 4-16Fused heteroaromatic ring pyridylcarbonyloxy, C 4-16 Fused heteroaromatic ring triazolylcarbonyloxy, C 2-6 Ester, pyridyl, phenyl, C 1-6 Alkoxy, C 2-20 Alkenyl, C 2-20 Alkynyl, C 2-8 Alkylcarbonyloxy or C 2-8 Alkoxycarbonyloxy, R 2 The substituents in the group are selected from deuterium, hydroxyl, amino, cyano, halogen, C 1-6 Alkyl, halogenated C 1-6 Alkyl, nitrooxy substituted C 1-6 Alkyl or C 1-6 One or more of alkoxy groups;

[0021] R 3 is hydrogen or C 1-6 alkyl.

[0022] In a preferred embodiment, Ar is a substituted or unsubstituted group: Wherein "*" is the connection site with C=O.

[0023] In a preferred embodiment, the compound of the present invention is selected from the compounds represented by the general formula (II), (III) or (IV),

[0024]

[0025] In a preferred embodiment, Y is O or NH.

[0026] In a preferred embodiment, R is C 3-6 Alkyl, substituted C 1-6 Alkyl, C 3-6 Cycloalkyl, substituted C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl or substituted C 3-6 Heterocycloalkyl; wherein the substituents in each group involved in R are selected from deuterium, cyano, nitro, halogen, C 1-5 Alkyl, C 1-5 Alkoxy or C 3-6 One or more of cycloalkyl groups.

[0027] In a preferred embodiment, R is C 3-6 Alkyl, substituted C 1-6 Alkyl, C 3-6 Cycloalkyl, substituted C 3-6 Cycloalkyl, tetrahydrofuran, substituted tetrahydrofuran, tetrahydrothiophene, substituted tetrahydrothiophene, tetrahydropyrrole or substituted tetrahydropyrrole; wherein the substituents in each group involved in R are selected from deuterium, cyano, nitro, halogen, C1-5 Alkyl, C 1-5 Alkoxy or C 3-6 One or more of cycloalkyl groups.

[0028] In a preferred embodiment, R is C 3-6 Alkyl, substituted C 1-3 Alkyl, C 3-6 Cycloalkyl or substituted C 3-6 Cycloalkyl, the substituent in the R group is selected from deuterium, halogen or C 3-6 Cycloalkyl.

[0029] In a preferred embodiment, R is C 3-6 Alkyl or C 3-6 Cycloalkyl.

[0030] In a preferred embodiment, R is n-propyl, isopropyl, n-butyl, isobutyl, cyclopropyl, cyclobutyl or cyclopentyl.

[0031] In a preferred embodiment, R 1 is a connecting bond or a substituted or unsubstituted C 1-4 Alkylene or substituted or unsubstituted C 4-12 Alkenylene, R 1 The substituents in the group are selected from deuterium, amino, cyano, halogen or C 1-4 One or more alkoxy groups.

[0032] In a preferred embodiment, R 2 is hydrogen, nitrooxy, carboxyl, or a substituted or unsubstituted dioxol-2-one, indazolyl, quinolyl, isoquinolyl, indolyl, benzofuranyl, purinyl, indazolylpyrazolylcarbonyloxy, quinolylpyrazolylcarbonyloxy, isoquinolylpyrazolylcarbonyloxy, indolylpyrazolylcarbonyloxy, benzofuranylpyrazolylcarbonyloxy, purinylpyrazolylcarbonyloxy, indazolylpyridinyl carbonyloxy, quinolylpyridylcarbonyloxy, isoquinolylpyridylcarbonyloxy, indolylpyridylcarbonyloxy, benzofuranylpyridylcarbonyloxy, purinylpyridylcarbonyloxy, indazolyltriazolylcarbonyloxy, quinolyltriazolylcarbonyloxy, isoquinolyltriazolylcarbonyloxy, indolyltriazolylcarbonyloxy, benzofuranyltriazolylcarbonyloxy, purinyltriazolylcarbonyloxy, C 2-6 Ester, pyridyl, phenyl, C 1-6 Alkoxy, C 6-20 Alkenyl, C 6-20 Alkynyl, C 2-8 Alkylcarbonyloxy or C 2-8 Alkoxycarbonyloxy, R 2 The substituents in the group are selected from deuterium, hydroxyl, amino, cyano, halogen, C 1-6Alkyl, halogenated C 1-6 Alkyl, nitrooxy substituted C 1-6 Alkyl or C 1-6 One or more alkoxy groups.

[0033] In a preferred embodiment, R 2 is hydrogen, nitrooxy, carboxyl or substituted or unsubstituted dioxol-2-one, indazolylpyrazolylcarbonyloxy, indazolylpyridinylcarbonyloxy, indazolyltriazolylcarbonyloxy, indolylpyrazolylcarbonyloxy, indolylpyridinylcarbonyloxy, indolyltriazolylcarbonyloxy, C 2-6 Ester, pyridyl, phenyl, C 1-6 Alkoxy, C 6-20 Alkenyl, C 2-8 Alkylcarbonyloxy or C 2-8 Alkoxycarbonyloxy, R 2 The substituents in the group are selected from deuterium, hydroxyl, amino, cyano, halogen, C 1-6 Alkyl, nitrooxy substituted C 1-6 Alkyl or C 1-6 One or more alkoxy groups.

[0034] In a preferred embodiment, R 3 is hydrogen, methyl, ethyl, n-propyl, isopropyl or butyl. In a preferred embodiment, the compound of the present invention can be selected from:

[0035]

[0036]

[0037] The present invention also includes a pharmaceutical composition, which uses the compound involved in this application or a pharmaceutically acceptable salt thereof as an active substance and is supplemented with pharmaceutically acceptable excipients.

[0038] The compound of the present invention or a pharmaceutically acceptable salt thereof can be used in the preparation of xanthine oxidase inhibitor drugs, in particular in the preparation of anti-gout drugs or anti-hyperuricemia drugs.

[0039] Unless otherwise specified, the groups indicated in the present invention have the following meanings:

[0040] "H", or hydrogen, refers to protium (1H), which is the main stable isotope of hydrogen.

[0041] "D", or "deuterium", refers to a stable isotope of hydrogen, also known as heavy hydrogen, with the element symbol D.

[0042] The term "halogen" refers to a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

[0043] "Hydroxyl" refers to an -OH group.

[0044] "Amino" refers to a -NH2 group.

[0045] "Alkyl" refers to a saturated aliphatic hydrocarbon group containing 1-10 carbon atoms, including straight-chain and branched groups (the numerical range mentioned in this application, such as "1-10", means that the group, in this case an alkyl group, can contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms). Alkyl groups containing 1-4 carbon atoms are called lower alkyl groups. When a lower alkyl group has no substituent, it is called an unsubstituted lower alkyl group. Alkyl groups can be C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl, C 1-2 Alkyl, C 2-3 Alkyl, C 2-4 Alkyl, etc. Specific alkyl groups include, but are not limited to, methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl or tert-butyl, etc. The alkyl group may be substituted or unsubstituted.

[0046] "Alkenyl" refers to a hydrocarbon group containing 2-30 carbon atoms and having one or more "C=C", including straight-chain and branched groups (the numerical range mentioned in this application, such as "2-10", refers to the group, in this case, alkenyl, which can contain 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms). Alkenyl can be selected from C 2-20 Alkenyl, C 2-18 Alkenyl, C 2-16 Alkenyl, C 2-14 Alkenyl, C 2-12 Alkenyl, C 4-14 Alkenyl, C 4-12 Specific alkenyl groups include, but are not limited to, vinyl, propenyl, allyl, butenyl, isobutenyl, tert-butenyl, wait.

[0047] "Alkoxy" refers to -O-(unsubstituted alkyl) and -O-(unsubstituted cycloalkyl) groups, which further refers to -O-(unsubstituted alkyl). The alkyl group can be C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl, C 1-2 Alkyl, C 2-3 Alkyl, C 2-4 Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, cyclopropyloxy, and the like.

[0048] "Dioxol-2-one" is group.

[0049] "Pyrazolyl" refers to Any one of .

[0050] "Triazolyl" includes 1,2,3-triazolyl, wherein "1,2,3-triazolyl" refers to

[0051] "Pyridyl" refers to Any one of .

[0052] "Fused heteroaromatic ring group" refers to an aromatic group containing two or more fused rings and heteroatoms, including but not limited to indazolyl, quinolyl, isoquinolyl, indolyl, benzofuranyl, purinyl, acridinyl, etc.

[0053] "Carboxyl" refers to a -COOH group.

[0054] "Ester" refers to a "-C(=O)-O-alkyl" group, in which the alkyl group can be C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl, C 1-2 Alkyl, C 2-3 Alkyl, C 2-4 Representative examples include, but are not limited to, methyl formate, ethyl formate, n-propyl formate, isopropyl formate, and the like. A substituted ester group means that a hydrogen atom in the ester group is replaced by a substituent, or that multiple hydrogen atoms in the ester group are replaced by the same or different substituents.

[0055] "Heterocycloalkyl" refers to a saturated cyclic group containing 3-10 ring atoms, wherein the ring atoms contain one or more heteroatoms selected from N, O, and S. The numerical range mentioned in this application, for example, "3-6", means that the group, which is a heterocycloalkyl at this time, can contain 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, etc., up to 6 carbon atoms as ring atoms. 3-8 Heterocycloalkyl, C 3-6 Heterocycloalkyl, C 3-5 Heterocycloalkyl, C 3-4 Heterocycloalkyl, C 3-9 Heterocycloalkyl, C 4-6Heterocycloalkyl, etc. Specific alkyl groups include, but are not limited to, tetrahydrofuran, tetrahydropyrrole, tetrahydrothiophene, 1,4-dioxane, oxospiro[3,3]heptyl, oxospiro[4,4]nonyl, oxospiro[5,5]undecyl, oxospiro[6,6]tridecyl, oxobicyclo[1,1,1]pentyl, oxobicyclo[2,2,2]octyl, oxobicyclo[3,2,1]octyl, azaspiro[3,3]heptyl, azaspiro[4,4]nonyl, azaspiro[5,5]undecyl, azaspiro[6,6]tridecyl, azabicyclo[1,1,1]pentyl, azabicyclo[2,2,2]octyl, or azabicyclo[3,2,1]octyl. Heterocycloalkyl groups may be substituted or unsubstituted.

[0056] “C 4-16 "Fused heteroaromatic ring pyrazolylcarbonyloxy" refers to a -OC(=O)-pyrazolyl-fused heteroaromatic ring group containing 4-16 carbon atoms, a specific example includes but is not limited to: indazolylpyrazolylcarbonyloxy Indolylpyrazolylcarbonyloxy

[0057] “C 2-8 "Alkoxycarbonyloxy" refers to an -OC(=O)-O-alkyl group containing 2 to 8 carbon atoms.

[0058] “C 2-8 "Alkylcarbonyloxy" refers to an -OC(=O)-alkyl group containing 2 to 8 carbon atoms.

[0059] “C 4-16 "Fused heteroaromatic ring pyridylcarbonyloxy" refers to a -OC(=O)-pyridyl-fused heteroaromatic ring group containing 4-16 carbon atoms, a specific example includes but is not limited to: indazolylpyridylcarbonyloxy Indolylpyridylcarbonyloxy

[0060] “C 4-16 "Fused heteroaromatic ring triazolylcarbonyloxy" refers to a -OC(=O)-triazolyl-fused heteroaromatic ring group containing 4-16 carbon atoms, a specific example includes but is not limited to: indazolyltriazolylcarbonyloxy Indolyltriazolylcarbonyloxy

[0061]

[0062] "Connecting bond" means that the groups at both ends are directly connected by covalent bonds. 1 -R 2 For example, when R 1 When it is a connecting bond, the group fragment is YR2 .

[0063] "Nitrooxy" refers to the -ONO2 group.

[0064] "Pharmaceutically acceptable salts" are salts of compounds of Formula (I) formed with organic or inorganic acids, meaning those salts that retain the biological effectiveness and properties of the parent compound. Such salts include, but are not limited to:

[0065] (1) Acid salts are obtained by reacting the free base of the parent compound with an inorganic acid or an organic acid, such as (but not limited to) hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid and perchloric acid, and organic acids such as (but not limited to) acetic acid, propionic acid, acrylic acid, oxalic acid, (D) or (L) malic acid, fumaric acid, maleic acid, hydroxybenzoic acid, γ-hydroxybutyric acid, methoxybenzoic acid, phthalic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, mandelic acid, succinic acid or malonic acid.

[0066] (2) Salts formed by the replacement of acidic protons in the parent compound by metal ions or coordination with organic bases, such as alkali metal ions, alkaline earth metal ions or aluminum ions, and organic bases such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, etc.

[0067] A "pharmaceutical composition" refers to a mixture of one or more compounds described herein, or their pharmaceutically acceptable salts and prodrugs, with other chemical components, such as pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.

[0068] The present invention further claims protection for a pharmaceutical composition comprising any one of the above-mentioned compounds, pharmaceutically acceptable salts or readily hydrolyzable prodrugs thereof and other pharmaceutically active ingredients.

[0069] The present invention also includes any of the above-mentioned compounds and pharmaceutically acceptable salts thereof, which can be formulated into any clinically or pharmaceutically acceptable dosage form using methods known in the art. For oral administration, conventional solid preparations such as tablets, capsules, pills, and granules can be prepared; oral liquid preparations such as oral solutions, oral suspensions, and syrups can also be prepared. Suitable fillers, binders, disintegrants, lubricants, and the like can be added to oral preparations. For parenteral administration, injections can be prepared, including injection solutions, sterile powders for injection, and concentrated solutions for injection. Injections can be produced using conventional methods in the pharmaceutical field. Additives may be omitted or added depending on the properties of the drug.

[0070] The compounds provided by this invention can significantly reduce serum uric acid levels in a hyperuricemia rat model and have potential application value in anti-gout and anti-hyperuricemia drugs. Because febuxostat can cause severe sudden cardiac death, severe nephrotoxicity, and severe hepatotoxicity, the compounds provided by this invention may have certain advantages in reducing drug toxicity and have good prospects for drug development. DETAILED DESCRIPTION

[0071] The present invention is further described below with reference to the following examples, but the protection scope of the present invention is not limited to the following examples.

[0072] Example 1: Synthesis of ethyl 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylate (2)

[0073]

[0074] Step A: A mixture containing 5-bromo-1H-indazole-3-carbonitrile (3.0 g, 13.5 mmol), isopropyl iodide (9.19 g, 54.1 mmol), cesium carbonate (8.80 g, 27.0 mmol) and DMF (50 mL) was stirred at 80°C for 1.5 hours. After cooling to room temperature, the insoluble matter was removed by filtration. Water (200 mL) was added and the mixture was extracted with ethyl acetate (80 mL × 3). The combined organic phase was washed with water (50 mL × 2) and saturated brine (50 mL) in sequence and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure and the product was purified by column chromatography (200-300 mesh silica gel, ethyl acetate:petroleum ether = 1:50-1:30) to obtain 5-bromo-1-isopropyl-1H-indazole-3-carbonitrile (1) (2.10 g). The yield was 58.9%. 1 H NMR (CDCl3, 400MHz) δ7.95 (d, J=1.2Hz, 1H), 7.55 (dd, J=1.2, 8.8Hz, 1H), 7.45 (d, J=8.8Hz, 1H), 4.93-4.87 (m, 1H), 1.61 (d, J=6.4Hz, 6H).

[0075] Step B: A mixture containing ethyl 1H-pyrazole-4-carboxylate (1.06 g, 7.56 mmol), compound 1 (1.0 g, 3.79 mmol), potassium carbonate (833 mg, 6.04 mmol), cuprous iodide (1.05 g, 5.51 mmol), (1S,2S)-1,2-diaminocyclohexane (432 mg, 3.78 mmol), and DMF (20 mL) was stirred at 110°C under nitrogen overnight. After cooling to room temperature, water (80 mL) was added and the mixture was extracted with ethyl acetate (40 mL x 3). The combined organic phases were washed with saturated brine (30 mL x 3) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, ethyl acetate:petroleum ether = 1:15-1:4) to give 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (2) (500 mg). The yield was 40.8%. 1 H NMR (DMSO-d6, 400MHz) δ9.31 (s, 1H), 8.47 (d, J = 1.2Hz, 1H), 8.23-8.16 (m, 3H), 5.2 9-5.22 (m, 1H), 4.29 (q, J=7.2Hz, 2H), 1.54 (d, J=7.2Hz, 6H), 1.33 (t, J=7.2Hz, 3H). MS (ESI, m / z): 324.1[M+H] + .

[0076] Example 2: Synthesis of 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid methyl ester (5)

[0077]

[0078] Step A: A mixture containing compound 2 (500 mg, 1.55 mmol), hydrated lithium hydroxide (623 mg, 14.8 mmol), water (1.5 mL), methanol (1.5 mL), and THF (1.5 mL) was stirred at 20°C for 2 hours. The solvent was partially evaporated under reduced pressure, water (8 mL) was added, and the pH was adjusted to 1-2 with 6M hydrochloric acid. The mixture was filtered and the filter cake was recrystallized from acetonitrile to give 1-(3-carbamoyl-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid (3) (300 mg). The yield was 61.8%. MS (ESI, m / z): 313.9 [M+H] + .

[0079] Step B: To a solution of compound 3 (300 mg, 0.958 mmol) in dichloromethane (5 mL) was added trifluoroacetic anhydride (906 mg, 4.31 mmol) and triethylamine (873 mg, 8.63 mmol) in an ice-water bath. After the addition was complete, the resulting mixture was stirred at room temperature overnight. Saturated brine (20 mL) was added, and the mixture was extracted with dichloromethane (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL x 2) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and then purified by preparative HPLC to obtain 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid (4). 1 H NMR (DMSO-d6, 400MHz) δ9.10 (s, 1H), 8.39 (d, J=1.6Hz, 1H), 8.21-8.12 (m, 2H), 8.05 (s, 1H), 5.22 (q, J=6.4Hz, 1H), 1.53 (d, J=6.4Hz, 6H). MS (ESI, m / z): 296.2[M+H] + .

[0080] Step C: A mixture containing compound 4 (250 mg, 0.847 mmol), iodomethane (192 mg, 1.35 mmol), potassium carbonate (235 mg, 1.70 mmol), and DMF (5 mL) was stirred at room temperature overnight. Water (20 mL) was added and the mixture was filtered. The filter cake was purified by column chromatography (200-300 mesh silica gel, eluted with dichloromethane:petroleum ether = 1:1) to obtain methyl 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylate (5). 1 H NMR (DMSO-d6, 400MHz) δ9.34 (s, 1H), 8.46 (d, J=1.6Hz, 1H), 8.23-8.16 (m, 3H), 5.28-5.22 (m, 1H), 3.83 (s, 3H), 1.54 (d, J=7.2Hz, 6H). MS (ESI, m / z): 310.1[M+H] + .

[0081] Example 3: Synthesis of 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl ester (6)

[0082]

[0083] A mixture containing compound 4 (150 mg, 0.508 mmol), 4-chloromethyl-5-methyl-1,3-dioxol-2-one (91 mg, 0.613 mmol), potassium carbonate (140 mg, 1.01 mmol), potassium iodide (110 mg, 0.663 mmol) and DMF (5 mL) was stirred at room temperature for 3 hours. Water (20 mL) was added and the mixture was filtered. The filter cake was purified by column chromatography (200-300 mesh silica gel, eluted with dichloromethane) to obtain 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl ester (6). 1 H NMR (DMSO-d6, 400MHz) δ9.47 (s, 1H), 8.58 (d, J=1.2Hz, 1H), 8.34-8.25 (m, 3H), 5.38-5.31 (m, 1H), 5.29 (s, 2H), 2.33 (s, 3H), 1.63 (d, J=6.8Hz, 6H). MS (ESI, m / z): 408.1[M+H] + .

[0084] Example 4: Synthesis of 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid isopropyl ester (7)

[0085]

[0086] A mixture containing compound 4 (250 mg, 0.847 mmol), isopropyl bromide (325 mg, 2.64 mmol), potassium carbonate (235 mg, 1.70 mmol), potassium iodide (190 mg, 1.14 mmol), and DMF (5 mL) was stirred at 30°C for 48 hours. Water (20 mL) was added and the mixture was filtered. The filter cake was purified by column chromatography (200-300 mesh silica gel, eluted with dichloromethane:petroleum ether = 1:1) to obtain 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid isopropyl ester (7) (190 mg). The yield was 66.5%. 1 HNMR (DMSO-d6, 400MHz) δ9.28 (s, 1H), 8.47 (d, J = 1.2Hz, 1H), 8.24-8.16 (m, 3H), 5 .29-5.22 (m, 1H), 5.16-5.10 (m, 1H), 1.54 (d, J=6.4Hz, 6H), 1.32 (d, J=6.4Hz, 6H). MS (ESI, m / z): 338.1[M+H] + .

[0087] Example 5: Synthesis of bis[1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid]propane-1,3-diester (8)

[0088]

[0089] A mixture containing compound 4 (200 mg, 0.677 mmol), 1,3-dibromopropane (68 mg, 0.337 mmol), potassium carbonate (187 mg, 1.35 mmol), potassium iodide (146 mg, 0.880 mmol), and DMF (5 mL) was stirred at 30°C for 48 hours. Water (20 mL) was added and the mixture was filtered. The filter cake was purified by column chromatography (200-300 mesh silica gel, eluted with dichloromethane:petroleum ether = 1:1) to obtain bis[1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid]propane-1,3-diester (8) (146 mg). The yield was 68.4%. 1 H NMR (DMSO-d6, 400MHz) δ9.20 (s, 2H), 8.24 (s, 2H), 8.12-8.05 (m, 6H), 5.23-5.1 6 (m, 2H), 4.45 (t, J=6.0Hz, 4H), 2.18 (t, J=6.0Hz, 2H), 1.53 (d, J=6.4Hz, 12H). MS (ESI, m / z): 631.1[M+H] + .

[0090] Example 6: Synthesis of [1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carbonyl]-L-valine methyl ester (9)

[0091]

[0092] A mixture containing compound 4 (200 mg, 0.677 mmol), L-valine methyl ester hydrochloride (136 mg, 0.811 mmol), diisopropylethylamine (219 mg, 1.69 mmol), HBTU (385 mg, 1.02 mmol), and DMF (5 mL) was stirred at room temperature overnight. Water (20 mL) was added, and the mixture was extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with saturated brine (15 mL × 3) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, eluted with petroleum ether:ethyl acetate:triethylamine = 100:10:1) to obtain [1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carbonyl]-L-valine methyl ester (9) (268 mg). The yield was 96.9%. 1H NMR (DMSO-d6, 400MHz) δ9.23 (s, 1H), 8.34-8.29 (m, 3H), 8.20-8.14 (m, 2H), 5.27-5.24 (m, 1H), 4.38-4.34 ( m, 1H), 3.67 (s, 3H), 2.18-2.13 (m, 1H), 1.55 (d, J=6.8Hz, 6H), 0.99 (d, J=6.8Hz, 3H), 0.94 (d, J=6.4Hz, 3H). MS (ESI, m / z): 409.2[M+H] + .

[0093] Example 7: Synthesis of 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid [(2R,3S)-3-amino-4-methoxy-4-oxobutan-2-yl] ester (11)

[0094]

[0095] Step A: A mixture containing compound 4 (200 mg, 0.677 mmol), Boc-L-threonine methyl ester (189 mg, 0.810 mmol), DCC (210 mg, 1.02 mmol), and dichloromethane (5 mL) was stirred at room temperature overnight. The insoluble matter was removed by filtration, and the filter cake was rinsed with dichloromethane (5 mL). The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, eluted with petroleum ether:ethyl acetate:triethylamine = 100:4:1) to obtain 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid [(2R,3S)-3-(Boc-amino)-4-methoxy-4-oxobutan-2-yl] ester (10) (340 mg). The yield was 99.8%.

[0096] Step B: A solution of compound 10 (340 mg, 0.666 mmol) and trifluoroacetic acid (0.3 mL) in dichloromethane (5 mL) was stirred at room temperature overnight. Water (20 mL) was added and the pH was adjusted to 7-8 with saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane (20 mL × 2), and the combined organic phases were washed with saturated brine (10 mL × 2) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the resulting product was recrystallized from ethyl acetate / petroleum ether to obtain [(2R,3S)-3-amino-4-methoxy-4-oxobutan-2-yl]-1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylate (11). 1H NMR (DMSO-d6, 400MHz) δ9.32 (s, 1H), 8.44 (s, 1H), 8.23-8.18 (m, 3H), 5.32-5.23 (m, 2H ), 3.61 (s, 3H), 3.55 (s, 1H), 2.06 (s, 2H), 1.54 (d, J = 6.4Hz, 6H), 1.35 (d, J = 6.4Hz, 3H). MS (ESI, m / z): 411.1[M+H] + .

[0097] Example 8: Synthesis of 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid (pyridin-2-yl)methyl ester (12)

[0098]

[0099] A mixture containing compound 4 (190 mg, 0.643 mmol), pyridine-2-methanol (84 mg, 0.770 mmol), DCC (210 mg, 0.969 mmol), DMAP (2 mg, 0.0163 mmol), and dichloromethane (5 mL) was stirred at room temperature overnight. Insoluble matter was removed by filtration, and the filter cake was rinsed with dichloromethane (5 mL). The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, dichloromethane:triethylamine = 100:1 elution) to obtain 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid (pyridin-2-yl)methyl ester (12) (75 mg). The yield was 30.2%. 1 H NMR (DMSO-d6, 400MHz) δ9.42 (s, 1H), 8.58 (d, J = 4.4Hz, 1H), 8.50 (d, J = 1.6Hz, 1H), 8.30 (s, 1H), 8.23 (d, J = 1.6Hz, 1H), 8.20 (s, 1H), 7.88-7.85 (m, 1H), 7.56 (d, J=8.0Hz, 1H), 7.39-7.36 (m, 1H), 5.40 (s, 2H), 5.29-5.23 (m, 1H), 1.54 (d, J=6.4Hz, 6H). MS (ESI, m / z): 387.1[M+H] + .

[0100] Example 9: Synthesis of 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid (3,7,11-trimethyldodecyl-2,6,10-trien-1-yl) ester (13)

[0101]

[0102] The experimental procedure for synthesizing compound 13 using compound 4 and 3,7,11-trimethyldodecyl-2,6,10-trien-1-ol as raw materials is shown in Example 8. 1 H NMR (DMSO-d6, 400MHz) δ9.29 (s, 1H), 8.47 (s, 1H), 8.23-8.15 (m, 3H), 5.44-5.40 (m, 1H), 5.29-5.22 (m , 1H), 5.10-5.00(m, 2H), 4.79-4.74(m, 2H), 2.11-1.89(m, 8H), 1.83-1.74(m, 6H), 1.58-1.52(m, 12H). MS (ESI, m / z): 500.3[M+H] + .

[0103] Example 10: Synthesis of 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid (3,7-dimethyloctane-2,6-dien-1-yl) ester (14)

[0104]

[0105] The experimental procedure for synthesizing compound 14 using compound 4 and 3,7-dimethyloctane-2,6-dien-1-ol as raw materials is shown in Example 8. 1 H NMR (DMSO-d6, 400MHz) δ9.31 (s, 1H), 8.47 (d, J = 1.6Hz, 1H), 8.23-8.16 (m, 3H), 5.44-5.41 (m, 1H), 5.28-5.23 (m, 1H), 5.22 -5.07 (m, 1H), 4.78 (d, J=6.8Hz, 2H), 2.10-2.03 (m, 4H), 1.74 (s, 3H), 1.63 (s, 3H), 1.57 (s, 3H), 1.55 (s, 3H), 1.53 (s, 3H). MS (ESI, m / z): 432.2[M+H] + .

[0106] Example 11: Synthesis of 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid (pivaloyloxy)methyl ester (15)

[0107]

[0108] A mixture containing compound 4 (120 mg, 0.406 mmol), chloromethyl pivalate (74 mg, 0.491 mmol), potassium carbonate (113 mg, 0.818 mmol), potassium iodide (88 mg, 0.530 mmol), and DMF (5 mL) was stirred at room temperature for 48 hours. Water (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 2). The combined organic phases were washed with saturated brine (20 mL × 2) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, dichloromethane:petroleum ether = 1:1 elution) to obtain 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid (pivaloyloxy)methyl ester (15). 1 HNMR (DMSO-d6, 400MHz) δ9.41 (s, 1H), 8.50 (d, J=1.2Hz, 1H), 8.26-8.17 (m, 3H), 5.94 (s, 2H), 5.29-5.22 (m, 1H), 1.54 (d, J=6.4Hz, 6H), 1.17 (s, 9H). MS (ESI, m / z): 410.1[M+H] + .

[0109] Example 12: Synthesis of methyl 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinate (19)

[0110]

[0111] Step A: To a mixture containing 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (8.40 g, 34.4 mmol), methyl 2-bromopyridine-4-carboxylate (9.0 g, 41.7 mmol), potassium carbonate (12.0 g, 87.0 mmol), dioxane (100 mL), and water (20 mL) was added [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.20 g, 1.47 mmol). After complete addition, the resulting mixture was stirred at 80°C under nitrogen for 3 hours. Cool to room temperature, filter, and rinse the filter cake with a small amount of ethyl acetate. Evaporate most of the solvent under reduced pressure, add ethyl acetate (500 mL), wash with saturated brine (100 mL), and dry over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, eluted with ethyl acetate:dichloromethane = 1:15) to give methyl 2-(1H-indazol-5-yl)isonicotinate (16). 1H NMR (DMSO-d6, 400MHz) δ13.25 (s, 1H), 8.90 (d, J=4.8Hz, 1H), 8.61 (s, 1H), 8.40 (s, 1H ), 8.23-8.19 (m, 2H), 7.79 (dd, J=1.2, 4.8Hz, 1H), 7.69 (d, J=8.8Hz, 1H), 3.99 (s, 3H). MS (ESI, m / z): 254.1[M+H] + .

[0112] Step B: To a solution of compound 16 (2.75 g, 10.9 mmol) in DMF (30 mL) were added cesium carbonate (7.08 g, 21.7 mmol) and iodine (5.50 g, 21.7 mmol). After complete addition, the resulting mixture was stirred at room temperature for 2 hours. Water (120 mL) and 2M sodium thiosulfate solution (20 mL) were added. The mixture was filtered and the filter cake was dissolved in ethyl acetate (300 mL). The insoluble matter was removed by filtration and then dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to obtain methyl 2-(3-iodo-1H-indazole-5-yl)isonicotinate (17) (3.90 g). The yield was 94.5%.

[0113] Step C: A mixture containing compound 17 (3.90 g, 10.3 mmol), potassium carbonate (1.70 g, 12.3 mmol), isopropyl bromide (1.90 g, 15.4 mmol), potassium iodide (340 mg, 2.05 mmol), and DMF (40 mL) was stirred at 60°C overnight. After cooling to room temperature, water (160 mL) was added and the mixture was extracted with ethyl acetate (100 mL x 2). The combined organic phases were washed sequentially with water (40 mL x 2) and saturated brine (40 mL), and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, ethyl acetate:petroleum ether = 1:10) to give methyl 2-(3-iodo-1-isopropyl-1H-indazol-5-yl)isonicotinate (18) (3.81 g). The yield was 87.8%.

[0114] Step D: A mixture containing compound 18 (3.81 g, 9.04 mmol), cuprous cyanide (1.14 g, 12.7 mmol), and DMF (30 mL) was stirred at 120°C overnight. After cooling to room temperature, ethyl acetate (100 mL) and water (100 mL) were added, and the insoluble matter was removed by filtration. The layers were separated, and the aqueous layer was extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed sequentially with water (40 mL × 2) and saturated brine (40 mL), and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, ethyl acetate:petroleum ether = 1:12 to 1:2) to obtain methyl 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinate (19) (1.0 g). The yield was 34.5%. 1 H NMR (DMSO-d6, 400MHz) δ8.92 (d, J=4.8Hz, 1H), 8.67 (s, 1H), 8.53 (s, 1H), 8.41 (d, J=8.8Hz, 1H), 8. 12 (d, J=8.8Hz, 1H), 7.83 (d, J=4.8Hz, 1H), 5.29-5.22 (m, 1H), 3.97 (s, 3H), 1.55 (d, J=6.8Hz, 6H). MS (ESI, m / z): 321.1[M+H] + .

[0115] Example 13: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid isopropyl ester (21)

[0116]

[0117] Step A: A mixture containing compound 19 (1.0 g, 3.12 mmol), 2 M sodium hydroxide solution (15 mL), methanol (5 mL), and THF (5 mL) was stirred at room temperature for 30 minutes. Water (20 mL) was added and the mixture was extracted with ethyl acetate (50 mL). The product was in the aqueous phase. The pH of the aqueous phase was adjusted to 5-6 with 2 M citric acid solution. Filtration afforded 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid (20) (688 mg). The yield was 72.0%. 1 H NMR (DMSO-d6, 400MHz) δ8.88 (d, J=4.8Hz, 1H), 8.65 (s, 1H), 8.50 (s, 1H), 8.40 (dd, J=1.6, 9.2H z, 1H), 8.13 (d, J=8.8Hz, 1H), 7.82 (d, J=8.8Hz, 1H), 5.30-5.23 (m, 1H), 1.58 (d, J=6.8Hz, 6H). MS (ESI, m / z): 307.3[M+H]+ .

[0118] The experimental procedure of step B is as in Example 4 to obtain isopropyl 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinate (21). 1 H NMR (DMSO-d6, 400MHz) δ8.91 (dd, J=0.8, 4.8Hz, 1H), 8.64 (d, J=0.8Hz, 1H), 8.48 (s, 1H), 8.41 (dd, J=1.6, 8.8Hz, 1H) , 8.12 (d, J=8.8Hz, 1H), 7.81 (dd, J=1.6, 4.8Hz, 1H), 5.29-5.20 (m, 2H), 1.56 (d, J=6.4Hz, 6H), 1.39 (d, J=6.4Hz, 6H). MS (ESI, m / z): 349.1[M+H] + .

[0119] Example 14: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl ester (22)

[0120]

[0121] The experimental procedure for synthesizing compound 22 using compound 20 and 4-chloromethyl-5-methyl-1,3-dioxol-2-one as raw materials is shown in Example 3. 1 H NMR (DMSO-d6, 400MHz) δ8.93 (d, J=5.2Hz, 1H), 8.67 (s, 1H), 8.53 (s, 1H), 8.40 (dd, J=1.6, 9.2Hz, 1H), 8.12 (d, J=9.2Hz, 1H), 7.85 (dd, J=1.6, 9.2Hz, 1H), 5.33 (s, 2H), 5.29-5.22 (m, 1H), 2.26 (s, 3H), 1.55 (d, J=6.8Hz, 6H). MS (ESI, m / z): 419.1[M+H] + .

[0122] Example 15: Synthesis of [2-(3-cyano-1-isopropyl-1H-indazol-5-yl)pyridine-4-carbonyl]-L-valine methyl ester (23)

[0123]

[0124] The experimental procedure for synthesizing compound 23 using compound 20 and L-valine methyl ester hydrochloride as raw materials is shown in Example 6. 1H NMR (DMSO-d6, 400MHz) δ9.10 (d, J=7.6Hz, 1H), 8.85 (d, J=4.8Hz, 1H), 8.65 ( s, 1H), 8.48 (s, 1H), 8.42 (d, J = 8.8Hz, 1H), 8.15 (d, J = 8.8Hz, 1H), 7.77 (d, J =5.2Hz, 1H), 5.28-5.23(m, 1H), 4.42-4.38(m, 1H), 3.70(s, 3H), 2.26-2.21 (m, 1H), 1.56 (d, J=6.8Hz, 6H), 1.03 (d, J=6.8Hz, 3H), 0.98 (d, J=6.8Hz, 3H). MS(ESI, m / z): 539.0[M+DMSO+ACN+H] + .

[0125] Example 16: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid (3,7,11-trimethyldodecyl-2,6,10-trien-1-yl) ester (24)

[0126]

[0127] The experimental procedure for synthesizing compound 24 using compound 20 and 3,7,11-trimethyldodecyl-2,6,10-trien-1-ol as raw materials is shown in Example 8. 1 H NMR (DMSO-d6, 400MHz) δ8.90 (d, J=4.8Hz, 1H), 8.63 (s, 1H), 8.48 (s, 1H), 8.38 (dd, J=1.6, 9.2Hz, 1H), 8.11 (d, J=9.2Hz, 1H), 7.80 (dd, J=1.6, 5. 2Hz, 1H), 5.49-5.46 (m, 1H), 5.29-5.22 (m, 1H), 5.11-5.07 (m, 2H), 4.92 -4.87(m, 2H), 2.12-1.88(m, 8H), 1.89-1.77(m, 6H), 1.64-1.52(m, 12H). MS (ESI, m / z): 511.3 [M+H] + .

[0128] Example 17: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid (3,7-dimethyloctane-2,6-dien-1-yl) ester (25)

[0129]

[0130] The experimental procedure for synthesizing compound 25 using compound 20 and 3,7-dimethyloctane-2,6-dien-1-ol as raw materials is shown in Example 8. 1 H NMR (DMSO-d6, 400MHz) δ8.90 (d, J=4.8Hz, 1H), 8.64 (s, 1H), 8.48 (s, 1H), 8.38 (dd, J=1.6, 9.2Hz, 1H), 8.11 (d, J=9.2Hz, 1H), 7.81 (dd, J=1.6, 5.2Hz, 1H), 5.49-5.47 (m, 1H), 5.26-5.24 (m, 1H), 5.08-5.07 (m, 1H), 4.92-4.90 (m, 2H), 2.09-2.04 (m, 4H), 1.77-1.57 (m, 15H). MS (ESI, m / z): 443.2[M+H] + .

[0131] Example 18: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid {1-[(ethoxycarbonyl)oxy]}ethyl ester (26)

[0132]

[0133] A mixture containing compound 20 (100 mg, 0.326 mmol), potassium carbonate (90 mg, 0.651 mmol), 1-chloroethyl ethyl carbonate (75 mg, 0.492 mmol), potassium iodide (70 mg, 0.422 mmol), and DMF (3 mL) was stirred at 40°C overnight. Water (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed sequentially with water (15 mL × 2) and saturated brine (15 mL), and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, eluted with petroleum ether:dichloromethane:triethylamine = 400:100:1) to obtain 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid {1-[(ethoxycarbonyl)oxy]}ethyl ester (26). 1H NMR (DMSO-d6, 400MHz) δ8.94 (d, J=5.2Hz, 1H), 8.67 (s, 1H), 8.52 (s, 1H), 8.40 (dd, J=1.6, 8.8Hz, 1H), 8.11 (d, J=8.8Hz, 1H), 7.84 (dd, J=1.6, 5 .2Hz, 1H), 6.96 (q, J=5.2Hz, 1H), 5.29-5.22 (m, 1H), 4.19 (q, J=6.8Hz, 2H), 1.65 (d, J=5.2Hz, 3H), 1.56 (d, J=6.8Hz, 6H), 1.24 (d, J=6.8Hz, 3H). MS (ESI, m / z): 423.1[M+H] + .

[0134] Example 19: Synthesis of (2-acetoxy)ethyl 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylate (27)

[0135]

[0136] The experimental procedure for synthesizing compound 27 using compound 4 and ethylene glycol monoacetate as raw materials is shown in Example 8. 1 HNMR (DMSO-d6, 400MHz) δ9.34 (s, 1H), 8.48 (d, J = 1.6Hz, 1H), 8.24-8.16 (m, 3H), 5.29- 5.23 (m, 1H), 4.47-4.45 (m, 2H), 4.35-4.33 (m, 2H), 2.06 (s, 3H), 1.55 (d, J=6.4Hz, 6H). MS (ESI, m / z): 382.5[M+H] + .

[0137] Example 20: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid (2-acetoxy)ethyl ester (28)

[0138]

[0139] The experimental procedure for synthesizing compound 28 using compound 20 and ethylene glycol monoacetate as raw materials is shown in Example 8. 1HNMR (DMSO-d6, 400MHz) δ8.93 (d, J=5.2Hz, 1H), 8.64 (s, 1H), 8.50 (s, 1H), 8.38 (dd, J=1.6, 8.8Hz, 1H), 8.13 (d, J=8.8Hz, 1H ), 7.83 (dd, J=1.6, 5.2Hz, 1H), 5.29-5.22 (m, 1H), 4.59-4.56 (m, 2H), 4.44-4.41 (m, 2H), 2.07 (s, 3H), 1.56 (d, J=6.8Hz, 6H). MS (ESI, m / z): 393.1[M+H] + .

[0140] Example 21: Synthesis of (2-methoxy)ethyl 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylate (29)

[0141]

[0142] The experimental procedure for synthesizing compound 29 using compound 4 and ethylene glycol monomethyl ether as raw materials is shown in Example 8. 1 HNMR (DMSO-d6, 400MHz) δ9.33 (s, 1H), 8.49 (d, J = 1.6Hz, 1H), 8.24-8.16 (m, 3H), 5.29-5.2 2 (m, 1H), 4.38 (t, J=4.4Hz, 2H), 3.65 (t, J=4.4Hz, 2H), 3.32 (s, 3H), 1.54 (d, J=6.8Hz, 6H). MS (ESI, m / z): 354.1[M+H] + Example 22: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid (2-methoxy)ethyl ester (30) and 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid (2-methoxy)ethyl ester hydrobromide (31)

[0143]

[0144] Step A: Using compound 20 and ethylene glycol monomethyl ether as raw materials, the experimental procedure for synthesizing compound 30 is shown in Example 8. 1H NMR (DMSO-d6, 400MHz) δ8.93 (d, J=4.8Hz, 1H), 8.66 (s, 1H), 8.51 (s, 1H), 8.39 (dd, J=1.6, 9.2Hz, 1H), 8.13 (d, J=9.2Hz, 1H), 7. 83 (dd, J=1.6, 5.2Hz, 1H), 5.29-5.23 (m, 1H), 4.51 (t, J=4.4Hz, 2H), 3.72 (t, J=4.4Hz, 2H), 3.33 (s, 3H), 1.55 (d, J=6.4Hz, 6H). MS (ESI, m / z): 365.1[M+H] + .

[0145] Step B: Add hydrogen bromide to a dichloromethane (10 mL) solution of compound 30 (48 mg, 0.132 mmol) to make the solution strongly acidic. Then, dichloromethane was evaporated and the solution was recrystallized from ethyl acetate / petroleum ether to obtain 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid (2-methoxy)ethyl ester hydrobromide (31). 1 H NMR (DMSO-d6, 400MHz) δ8.93 (d, J=4.8Hz, 1H), 8.65 (s, 1H), 8.51 (s, 1H), 8.39 (dd, J=1.6, 8.8Hz, 1H), 8.13 (d, J=8.8Hz, 1H) , 7.83 (dd, J=1.6, 5.2Hz, 1H), 5.29-5.23 (m, 1H), 4.52-4.50 (m, 2H), 3.74-3.71 (m, 2H), 3.33 (s, 3H), 1.55 (d, J=6.4Hz, 6H). MS (ESI, m / z): 365.1[M+H] + .

[0146] Example 23: Synthesis of cinnamyl 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylate (32)

[0147]

[0148] For the experimental procedure of synthesizing compound 32 using compound 4 and cinnamyl alcohol as raw materials, see Example 8. 1H NMR (DMSO-d6, 400MHz) δ9.38 (s, 1H), 8.49 (d, J=1.6Hz, 1H), 8.26-8.17 (m, 3H), 7.52-7.50 (m, 2H), 7.39-7.29 (m, 3H), 6.81 (d, J=16.0Hz, 1H), 6.54-6.46 (m, 1H), 5.29-5.22 (m, 1H), 4.95 (d, J=5.6Hz, 2H), 1.54 (d, J=6.8Hz, 6H). MS (ESI, m / z): 412.1[M+H] + .

[0149] Example 24: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)cinnamyl isonicotinate (33)

[0150]

[0151] For the experimental procedure of synthesizing compound 33 using compound 20 and cinnamyl alcohol as raw materials, see Example 8. 1 H NMR (DMSO-d6, 400MHz) δ8.93 (d, J=4.8Hz, 1H), 8.68 (s, 1H), 8.56 (s, 1H), 8.39 (dd, J=1.6, 8.8Hz, 1H), 8.12 (d, J=8.8Hz, 1H), 7.84 (dd, J=1.6, 4.8Hz, 1 H), 7.54-7.52 (m, 2H), 7.39-7.30 (m, 3H), 6.86 (d, J=16.4Hz, 1H), 6.60-6. 52 (m, 1H), 5.29-5.22 (m, 1H), 5.08 (d, J = 5.6Hz, 2H), 1.55 (d, J = 6.8Hz, 6H). MS (ESI, m / z): 423.1 [M+H] + .

[0152] Example 25: Synthesis of (1-isobutyryloxy)ethyl 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylate (34)

[0153]

[0154] For the experimental procedure of synthesizing compound 34 using compound 4 and 1-chloroethyl isobutyrate as raw materials, see Example 18. 1HNMR (DMSO-d6, 400MHz) δ9.36 (s, 1H), 8.48 (s, 1H), 8.23-8.16 (m, 3H), 6.98 (q, J=5.6 Hz, 1H), 5.29-5.22 (m, 1H), 2.62-2.55 (m, 1H), 1.57-1.54 (m, 9H), 1.12-1.10 (m, 6H). MS (ESI, m / z): 410.1[M+H] + .

[0155] Example 26: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid (1-isobutyryloxy)ethyl ester (35)

[0156]

[0157] For the experimental procedure of synthesizing compound 35 using compound 20 and 1-chloroethyl isobutyrate as raw materials, see Example 18. 1 H NMR (DMSO-d6, 400MHz) δ8.94 (d, J=4.8Hz, 1H), 8.66 (s, 1H), 8.50 (s, 1H), 8.39 (dd, J=1.6, 9.2Hz, 1H), 8.12 (d, J=9.2Hz, 1H), 7.83 (dd, J=1.2, 4.8Hz, 1H), 7.04 (q, J=5.2Hz, 1H), 5.29 -5.22 (m, 1H), 2.65-2.58 (m, 1H), 1.63 (d, J=5.2Hz, 3H), 1.56 (d, J=6.4Hz, 6H), 1.12 (d, J=6.8Hz, 6H). MS (ESI, m / z): 421.2[M+H] + .

[0158] Example 27: Synthesis of bis[2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid]propane-1,3-diester (36)

[0159]

[0160] For the experimental procedure of synthesizing compound 36 using compound 20 and 1,3-dibromopropane as raw materials, see Example 5. 1HNMR (DMSO-d6, 400MHz) δ8.72 (d, J=4.8Hz, 2H), 8.32-8.31 (m, 4H), 8.17 (dd, J=1.2, 8.8Hz, 2H), 7.98 (d, J=8.8Hz, 2 H), 7.71 (d, J=4.8Hz, 2H), 5.22-5.15 (m, 2H), 4.62 (t, J=6.0Hz, 4H), 2.33 (t, J=6.0Hz, 2H), 1.53 (d, J=6.4Hz, 12H). MS (ESI, m / z): 653.2[M+H] + .

[0161] Example 28: Synthesis of 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid [4-(nitrooxy)]butyl ester (38)

[0162]

[0163] Step A: A mixture containing 4-bromobutyl acetate (1.0 g, 5.13 mmol), silver nitrate (1.30 g, 7.65 mmol) and acetonitrile (15 mL) was stirred under reflux overnight in the dark. After cooling to room temperature, the insoluble matter was removed by filtration. Water (60 mL) was added and the mixture was extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and then 2M sodium hydroxide solution (2.5 mL) and methanol (5 mL) were added to the residue. After the addition was complete, the resulting mixture was stirred at room temperature for 2 hours. Water (20 mL) was added and the mixture was extracted with ethyl acetate (20 mL × 2). The combined organic phases were washed with saturated brine (10 mL) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, petroleum ether:ethyl acetate = 5:1) to obtain 4-hydroxybutyl nitrate (37) (400 mg). The yield was 57.5%.

[0164] Step B: A mixture containing compound 4 (80 mg, 0.272 mmol), compound 37 (40 mg, 0.296 mmol), DCC (84 mg, 0.407 mmol), DMAP (4 mg, 0.0327 mmol) and dichloromethane (5 mL) was stirred at room temperature overnight. The insoluble matter was removed by filtration. The solvent was evaporated under reduced pressure, and the product was purified by column chromatography (200-300 mesh silica gel, petroleum ether:ethyl acetate = 10:1-10:3) to obtain 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid [4-(nitrooxy)]butyl ester (38). 1H NMR (DMSO-d6, 400MHz) δ9.31 (s, 1H), 8.46 (s, 1H), 8.23-8.16 (m, 3H), 5.29-5.22 (m, 1H) , 4.61 (t, J=6.0Hz, 2H), 4.29 (t, J=6.0Hz, 2H), 1.87-1.80 (m, 4H), 1.54 (d, J=6.4Hz, 6H). MS (ESI, m / z): 413.3[M+H] + .

[0165] Example 29: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid [4-(nitrooxy)]butyl ester (39)

[0166]

[0167] The experimental procedure for synthesizing compound 39 using compound 20 and compound 37 as raw materials refers to step B in Example 28. 1 H NMR (DMSO-d6, 400MHz) δ8.91 (d, J=4.8Hz, 1H), 8.63 (s, 1H), 8.49 (s, 1H), 8.38 ( dd, J=1.6, 9.2Hz, 1H), 8.11 (d, J=9.2Hz, 1H), 7.83 (dd, J=1.2, 4.8Hz, 1H), 5.29 -5.22 (m, 1H), 4.62 (t, J=6.0Hz, 2H), 4.42 (t, J=6.0Hz, 2H), 1.89-1.86 (m, 4H), 1.56 (d, J=6.4Hz, 6H). MS (ESI, m / z): 424.0[M+H] + Example 30: Synthesis of [3-(nitrooxy)methyl]phenyl 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylate (41)

[0168]

[0169] Step A: A mixture containing m-hydroxybenzyl bromide (500 mg, 2.67 mmol), silver nitrate (500 mg, 2.94 mmol) and acetonitrile (5 mL) was stirred in an ice-water bath in the dark for 5 hours. The insoluble matter was removed by filtration. Water (20 mL) was added and the mixture was extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (10 mL) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure and the product was purified by column chromatography (200-300 mesh silica gel, petroleum ether:ethyl acetate = 35:1) to give 3-hydroxybenzyl nitrate (40) (230 mg). The yield was 50.9%.

[0170] The experimental procedure of step B was similar to that of step B in Example 28 to obtain 1-(3-cyano-1-isopropyl-1H-indazol-5-yl)-1H-pyrazole-4-carboxylic acid [3-(nitrooxy)methyl]phenyl ester (42). 1 H NMR (DMSO-d6, 400MHz) δ9.58 (s, 1H), 8.53 (d, J = 1.6Hz, 1H), 8.40 (s, 1H), 8.28-8.19 (m, 2H), 7.57-7.5 3 (m, 1H), 7.44-7.42 (m, 2H), 7.37-7.35 (m, 1H), 5.64 (s, 2H), 5.30-5.23 (m, 1H), 1.55 (d, J=6.4Hz, 6H). MS (ESI, m / z): 447.0[M+H] + .

[0171] Example 31: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid [3-(nitrooxy)methyl]phenyl ester (42)

[0172]

[0173] The experimental procedure for synthesizing compound 42 using compound 20 and compound 40 as raw materials refers to step B in Example 28. 1 H NMR (DMSO-d6, 400MHz) δ9.00 (d, J=5.2Hz, 1H), 8.73 (s, 2H), 8.46 (dd, J=1.6, 8.8Hz, 1H), 8.14 (d, J=8.8Hz, 1H), 8.01 (dd, J=1. 6, 5.2Hz, 1H), 7.61-7.57 (m, 1H), 7.53-7.52 (m, 1H), 7.48-7.45 (m, 2H), 5.66 (s, 2H), 5.30-5.23 (m, 1H), 1.56 (d, J=6.8Hz, 6H). MS (ESI, m / z): 457.9[M+H] + .

[0174] Example 32: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid [2-(nitrooxy)]ethyl ester (44)

[0175]

[0176] For the experimental procedure of synthesizing compound 44 using 2-iodoethanol and compound 20 as raw materials, see Example 28. 1H NMR (DMSO-d6, 400MHz) δ8.91 (d, J=4.8Hz, 1H), 8.63 (s, 1H), 8.50 (s, 1H), 8.37 ( dd, J=1.6, 8.8Hz, 1H), 8.11 (d, J=8.8Hz, 1H), 7.81 (dd, J=1.2, 4.8Hz, 1H), 5.27 -5.20 (m, 1H), 4.94-4.92 (m, 2H), 4.69-4.67 (m, 2H), 1.53 (d, J=6.8Hz, 6H). MS (ESI, m / z): 396.0[M+H] + .

[0177] Example 33: Synthesis of 2-(3-cyano-1-isopropyl-1H-indazol-5-yl)isonicotinic acid [3-(nitrooxy)]propyl ester (46)

[0178]

[0179] For the experimental procedure of synthesizing compound 46 using 3-bromo-1-propanol and compound 20 as raw materials, see Example 28. 1 HNMR (DMSO-d6, 400MHz) δ8.91 (d, J=5.2Hz, 1H), 8.63 (s, 1H), 8.49 (s, 1H), 8.38 (dd, J=1.6, 8.8Hz, 1H), 8.11 (d, J=8.8Hz, 1H), 7.83 (dd, J=1.2, 4.8Hz, 1H), 5.28-5.18 (m, 1H), 4.56 (t, J=6.4Hz, 2H), 4.43 (t, J=6.4Hz, 2H), 1.91-1.87 (m, 2H), 1.54 (d, J=6.8Hz, 6H). MS (ESI, m / z): 468.5[M+ACN+Na] + .

[0180] Example 34: Experimental study on the effect of compound 22 on the treatment of hyperuricemia in rats

[0181] 1. Experimental Materials

[0182] (1) Test drug

[0183] Compound 22 was a light yellow powder, which was ground with 0.5% CMC-Na before use and prepared into a suspension of corresponding concentration for oral administration.

[0184] Febuxostat was purchased from Sigma and ground with 0.5% CMC-Na before use to prepare a suspension of corresponding concentration for oral gavage.

[0185] (2) Animals and breeding

[0186] a. Animal species and source

[0187] SD rats, SPF grade, 36 males, weighing 180-200 g, were purchased from Shanghai Slake Laboratory Animal Co., Ltd., production license number: SCXK (Beijing) 2019-0010, quality certificate number: 110324221100913432.

[0188] b. Rearing conditions

[0189] All rats were housed in independent ventilation cages with an air cleanliness level of 10000, a laboratory temperature of 26±2°C, a relative humidity of 60% to 80%, air exchanges of 10-15 times per hour, and a photoperiod of 12 (day) / 12 (night) hours, with 3 rats per cage.

[0190] Feed: Complete pelleted feed for rats was purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd., and its quality complies with GB14924.1-2001 “General Quality Standard for Compound Feeds for Laboratory Animals”.

[0191] Bedding: Sterile granular bedding was purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd.

[0192] Drinking water: Purified water is available for drinking freely after acidification.

[0193] (3) Main instruments and equipment

[0194] Varioskan LUX multifunctional microplate reader was purchased from Thermo Fisher Scientific, USA; BS210S precision electronic balance (0.1 mg–10 g) was purchased from Sartorius, Germany; FEJ-200 electronic balance (0.1–200 g) was purchased from Fuzhou Furihengzhibao Electronics Co., Ltd.; and Pacific TII+Genpure XCAD PLUS UV / TOC / UF pure water ultrapure water system was purchased from Thermo Fisher Scientific, USA.

[0195] (4) Main reagents

[0196] Uric acid detection kit (phosphotungstic acid reduction method), batch number: 20220305, was purchased from Nanjing Jiancheng Bioengineering Institute; potassium oxonate, product number 00164, batch number GR4VI-RK, was purchased from Tokyo Chemical Industry Co., Ltd. (TCI), Japan; sodium carboxymethyl cellulose (CMC-Na), batch number 20170810, chemically pure, was purchased from Sinopharm Chemical Reagent Co., Ltd.

[0197] 2. Experimental Methods

[0198] (1) Grouping

[0199] Thirty-six male SD rats weighed approximately 200-230 g after one week of acclimation. They were randomly divided into six groups, stratified by body weight, with six rats in each group: (1) normal group (0.5% CMC-Na), (2) model group (0.5% CMC-Na), (3) febuxostat 1 mg / kg, (4) febuxostat 2 mg / kg, (5) compound 22, 1.45 mg / kg, and (6) compound 22, 2.9 mg / kg. Each drug was prepared into a suspension of the corresponding concentration, and the administration volume was 0.5 mL / 100 g.

[0200] (2) Model establishment, dosing regimen, and testing indicators

[0201] Rats in each group were purchased and acclimated to the rat population. After fasting for 12 hours, rats were treated with potassium oxonate at a dose of 300 mg / kg intraperitoneally to establish a rat model. 0.5 hours after model establishment, each test drug group received a single oral gavage. Blood was collected from the retroorbital venous plexus before potassium oxonate injection and 1, 3, and 5 hours after potassium oxonate injection. The blood was centrifuged at 3500 rpm for 10 minutes, and 30 μL of serum was collected to measure uric acid levels at each time point.

[0202] (3) Data processing and statistical methods

[0203] All experimental measurement data were expressed as (mean) ± SD), and the significance was examined by ANOVA-Dunnett T test between groups, with P < 0.05 as the significance index and P < 0.01 as the extremely significant index.

[0204] 3. Experimental Results

[0205] The results are shown in Table 1. Compared with the solvent group, the serum uric acid levels in the potassium oxonate model group were significantly increased 1, 3, and 5 hours after modeling (P < 0.05). Compared with the model group at the same time point, both 1 mg / kg and 2 mg / kg of febuxostat significantly reduced the serum uric acid levels 1, 3, and 5 hours after modeling (P < 0.01). Compared with the model group at the same time point, the 1.45 mg / kg group of compound 22 significantly reduced the serum uric acid level 1 hour after modeling (P < 0.05). The 2.9 mg / kg group of compound 22 significantly reduced the serum uric acid levels 1 and 5 hours after modeling (P < 0.05).

[0206] Table 1. Effects of drug administration on serum uric acid levels in rats with hyperuricemia induced by potassium oxonate

[0207]

[0208] Note: # P<0.05, ##P<0.01, compared with the solvent group at the same time point; *P<0.05, **P<0.01, compared with the model group at the same time point.

[0209] Example 35: Experimental study on the effects of compounds 13, 38 and 41 on the treatment of hyperuricemia in rats

[0210] 1. Experimental Materials

[0211] (1) Test drug

[0212] Compound 13 was a light yellow powder, and compounds 38 and 41 were off-white powders. They were ground with 0.5% CMC-Na before use and prepared into a 0.4 mg / mL suspension for oral administration.

[0213] Febuxostat was purchased from Sigma and ground with 0.5% CMC-Na before use to prepare a 0.4 mg / mL suspension for oral gavage.

[0214] (2) Animals and breeding

[0215] a. Animal species and source

[0216] SD rats, SPF grade, 36 males, weighing 180-200 g, were purchased from Shanghai Slake Laboratory Animal Co., Ltd., production license number: SCXK (Beijing) 2019-0010, quality certificate number: 110324221100913432.

[0217] b. Rearing conditions

[0218] All rats were housed in independent ventilation cages with an air cleanliness level of 10000, a laboratory temperature of 26±2°C, a relative humidity of 60% to 80%, air exchanges of 10-15 times per hour, and a photoperiod of 12 (day) / 12 (night) hours, with 3 rats per cage.

[0219] Feed: Complete pelleted feed for rats was purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd., and its quality complies with GB14924.1-2001 “General Quality Standard for Compound Feeds for Laboratory Animals”.

[0220] Bedding: Sterile granular bedding was purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd.

[0221] Drinking water: Purified water is available for drinking freely after acidification.

[0222] (3) Main instruments and equipment

[0223] Varioskan LUX multifunctional microplate reader was purchased from Thermo Fisher Scientific, USA; BS210S precision electronic balance (0.1 mg–10 g) was purchased from Sartorius, Germany; FEJ-200 electronic balance (0.1–200 g) was purchased from Fuzhou Furihengzhibao Electronics Co., Ltd.; and Pacific TII+Genpure XCAD PLUS UV / TOC / UF pure water ultrapure water system was purchased from Thermo Fisher Scientific, USA.

[0224] (4) Main reagents

[0225] Uric acid detection kit (phosphotungstic acid reduction method), batch number: 20230224, was purchased from Nanjing Jiancheng Bioengineering Institute; potassium oxonate, product number 00164, batch number T6GKM-TA, was purchased from Tokyo Chemical Industry Co., Ltd. (TCI), Japan; sodium carboxymethyl cellulose (CMC-Na), batch number 20170810, chemically pure, was purchased from Sinopharm Chemical Reagent Co., Ltd.

[0226] 2. Experimental Methods

[0227] (1) Grouping

[0228] Thirty-six male SD rats weighed approximately 220-240 g after one week of acclimation. They were randomly divided into six groups, stratified by body weight, with six rats in each group: (1) normal group (0.5% CMC-Na), (2) model group (0.5% CMC-Na), (3) febuxostat 2 mg / kg, (4) compound 13, 2 mg / kg, (5) compound 38, 2 mg / kg, and (6) compound 41, 2 mg / kg. Each drug was prepared into a suspension of the corresponding concentration, and the administration volume was 0.5 mL / 100 g.

[0229] (2) Model establishment, dosing regimen, and testing indicators

[0230] Rats in each group were purchased and acclimated to the rat population. After fasting for 12 hours, rats were treated with potassium oxonate at a dose of 300 mg / kg per ip to establish the model. 0.5 hours after model establishment, each test drug group received the drug once via gavage. Drug administration continued for three consecutive days. On the third day, blood was collected from the retroorbital venous plexus before potassium oxonate injection and 1, 3, and 5 hours after potassium oxonate injection. The blood was centrifuged at 3500 rpm for 10 minutes, and 30 μL of serum was collected to determine uric acid levels at each time point.

[0231] (3) Data processing and statistical methods

[0232] All experimental measurement data were expressed as (mean) ± SD), and the significance was examined by ANOVA-Dunnett T test between groups, with P < 0.05 as the significance index and P < 0.01 as the extremely significant index.

[0233] 3. Experimental Results

[0234] The results are shown in Table 2. Compared with the solvent group, the serum uric acid level in the potassium oxonate model group was significantly increased 1, 3, and 5 hours after modeling (P < 0.05). Compared with the model group at the same time point, the febuxostat group significantly reduced the serum uric acid level 1, 3, and 5 hours after modeling (P < 0.01). Compared with the model group at the same time point, compound 13 significantly reduced the serum uric acid level 1, 3, and 5 hours after modeling (P < 0.01 or P < 0.05). Compound 38 significantly reduced the serum uric acid level 1, 3, and 5 hours after modeling (P < 0.01 or P < 0.05). Compound 41 significantly reduced the serum uric acid level 3 and 5 hours after modeling (P < 0.01).

[0235] Table 2. Effects of drug administration on serum uric acid levels in rats with hyperuricemia induced by potassium oxonate

[0236]

[0237] Note: ## P<0.01, compared with the solvent group at the same time point; *P<0.05, **P<0.01, compared with the model group at the same time point.

[0238] Example 36: Pharmacokinetics of the compound in SD rats

[0239] 1. Experimental Materials

[0240] (1) Test drug

[0241] Preparation of compound stock solution: Weigh appropriate amount of compound solid powder, add a certain amount of DMSO, vortex and sonicate to obtain 10 mg / mL stock solution.

[0242] Preparation of test compounds for oral administration: Pipette appropriate amount of compound stock solution, add a certain amount of Solutol HS15 solution, vortex for 1 minute, then add a certain amount of physiological saline, mix thoroughly to obtain a 1 mg / mL solution.

[0243] Preparation of test compounds for intravenous injection: Pipette an appropriate amount of compound stock solution, add a certain amount of Solutol HS15 solution, vortex for 1 minute, then add a certain amount of normal saline, mix thoroughly to obtain a 0.5 mg / mL solution.

[0244] (2) Experimental animals

[0245] Male Sprague-Dawley rats, SPF grade, 6–8 weeks old, were purchased from JH Laboratory Animal Co., Ltd. License numbers: SCXK(SH)2017-0012, SCXK(SH)2022-0009, Certificate numbers: 20170012022154, 20220009005149.

[0246] 2. Experimental Methods

[0247] (1) Dosage and method of administration

[0248] The experimental animals were fasted overnight before oral administration and were fed again 4 hours after administration. Water was available to the animals during this period. Two groups were set up for each test compound: intravenous administration and oral administration. The specific dosage and method of administration are shown in Table 3 below.

[0249] Table 3. Dosage and administration method of the compounds to SD rats

[0250] Group Dosage (mg / kg) Dosing volume (mL / kg) Concentration (mg / mL) Route of administration Intravenous administration group 1 2 0.5 intravenous injection Oral administration group 10 10 1 oral

[0251] (2) Experimental operation

[0252] Blood samples (150 μL / sample) were collected from the jugular vein of SD rats before administration and at 5 minutes (intravenous administration only), 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, and 24 hours after administration. The samples were placed in centrifuge tubes containing the anticoagulant sodium heparin and centrifuged at 2000 g for 5 minutes at 4°C to separate plasma. The plasma samples were analyzed by LC / MS / MS to determine the concentration of each test compound.

[0253] (3) Pharmacokinetic analysis

[0254] The non-compartmental model parameters are given by Calculated using Professional software.

[0255] 3. Experimental Results

[0256] The pharmacokinetic parameters of the test compounds in SD rats obtained according to the above method are shown in Table 4. The pharmacokinetic parameters of the compounds of the present invention are good and the bioavailability is high.

[0257] Table 4. Pharmacokinetic parameters of the compounds in SD rats after oral or intravenous administration

[0258]

[0259]

Claims

1. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof, in, R is C 1-6 Alkyl, substituted C 1-6 Alkyl, C 3-6 Cycloalkyl, substituted C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl or substituted C 3-6 Heterocycloalkyl; wherein the substituents in each group involved in R are selected from deuterium, cyano, nitro, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or C 3-6 one or more of heterocycloalkyl; Ar is a substituted or unsubstituted group: The substituents in the Ar group are selected from deuterium, hydroxyl, halogen, C 1-4 Alkyl or C 1-4 one or more of alkoxy groups; Y is O or NR 3 , R 1 is substituted or unsubstituted C 1-6 Alkylene or substituted or unsubstituted C 2-12 Alkenylene, R 1 The substituents in the group are selected from deuterium, hydroxyl, amino, cyano, halogen, C 1-4 Alkyl or C 1-4 one or more of alkoxy groups; R 2 is hydrogen, carboxyl or the following substituted or unsubstituted groups: dioxol-2-one, C 4-12 Condensed heteroaromatic ring group, C 4-16 Fused heteroaromatic ring pyrazolylcarbonyloxy, C 4-16 Fused heteroaromatic ring pyridylcarbonyloxy, C 4-16 Fused heteroaromatic ring triazolylcarbonyloxy, C 2-6 Ester, pyridyl, phenyl, C 1-6 Alkoxy, C 2-20 Alkenyl, C 2-20 Alkynyl, C 2-8 Alkylcarbonyloxy or C 2-8 Alkoxycarbonyloxy, R 2 The substituents in the group are selected from deuterium, hydroxyl, amino, cyano, halogen, C 1-6 Alkyl or C 1-6 One or more of alkoxy groups; R 3 is hydrogen or C 1-6 alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the compounds represented by general formula (II), (III) or (IV), 3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y is O or NH, and R is C 3-6 Alkyl, substituted C 1-6 Alkyl, C 3-6 Cycloalkyl, substituted C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl or substituted C 3-6 Heterocycloalkyl; wherein the substituents in each group involved in R are selected from deuterium, cyano, nitro, halogen, C 1-5 Alkyl, C 1-5 Alkoxy or C 3-6 One or more of cycloalkyl groups.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R is C 3-6 Alkyl, substituted C 1-6 Alkyl, C 3-6 Cycloalkyl, substituted C 3-6 Cycloalkyl, tetrahydrofuran, substituted tetrahydrofuran, tetrahydrothiophene, substituted tetrahydrothiophene, tetrahydropyrrole or substituted tetrahydropyrrole; wherein the substituents in each group involved in R are selected from deuterium, cyano, nitro, halogen, C 1-5 Alkyl, C 1-5 Alkoxy or C 3-6 One or more of cycloalkyl groups.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 is substituted or unsubstituted C 1-4 Alkylene or substituted or unsubstituted C 4-12 Alkenylene, R 1 The substituents in the group are selected from deuterium, amino, cyano, halogen or C 1-4 One or more alkoxy groups.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 2 is hydrogen, carboxyl, or a substituted or unsubstituted dioxol-2-one group, indazolyl, quinolyl, isoquinolyl, indolyl, benzofuranyl, purinyl, indazolylpyrazolylcarbonyloxy, quinolylpyrazolylcarbonyloxy, isoquinolylpyrazolylcarbonyloxy, indolylpyrazolylcarbonyloxy, benzofuranylpyrazolylcarbonyloxy, purinylpyrazolylcarbonyloxy, indazolylpyridinylcarbonyloxy yl, quinolylpyridylcarbonyloxy, isoquinolylpyridylcarbonyloxy, indolylpyridylcarbonyloxy, benzofuranylpyridylcarbonyloxy, purinylpyridylcarbonyloxy, indazolyltriazolylcarbonyloxy, quinolyltriazolylcarbonyloxy, isoquinolyltriazolylcarbonyloxy, indolyltriazolylcarbonyloxy, benzofuranyltriazolylcarbonyloxy, purinyltriazolylcarbonyloxy, C 2-6 Ester, pyridyl, phenyl, C 1-6 Alkoxy, C 6-20 Alkenyl, C 6-20 Alkynyl, C 2-8 Alkylcarbonyloxy or C 2-8 Alkoxycarbonyloxy, R 2 The substituents in the group are selected from deuterium, hydroxyl, amino, cyano, halogen, C 1-6 Alkyl or C 1-6 One or more alkoxy groups.

7. The compound according to claim 5 or a pharmaceutically acceptable salt thereof, wherein R 2 is hydrogen, carboxyl or the following substituted or unsubstituted groups: dioxol-2-one, indazolylpyrazolylcarbonyloxy, indazolylpyridinylcarbonyloxy, indazolyltriazolylcarbonyloxy, indolylpyrazolylcarbonyloxy, indolylpyridinylcarbonyloxy, indolyltriazolylcarbonyloxy, C 2-6 Ester, pyridyl, phenyl, C 1-6 Alkoxy, C 6-20 Alkenyl, C 2-8 Alkylcarbonyloxy or C 2-8 Alkoxycarbonyloxy, R 2 The substituents in the group are selected from deuterium, hydroxyl, amino, cyano, halogen, C 1-6 Alkyl or C 1-6 One or more alkoxy groups.

8. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

9. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof as an active substance and pharmaceutically acceptable excipients.

10. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of an anti-gout drug or an anti-hyperuricemia drug.

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