FTO inhibitor and preparation method and application thereof

Abstract

The invention provides an FTO (fluorine-doped tin oxide) inhibitor as well as a preparation method and application thereof, and particularly discloses 2-(substituted benzene miscellaneous group) aromatic formic acid as shown in the following general formula (I), derivative compounds of the 2-(substituted benzene miscellaneous group) aromatic formic acid and pharmaceutically acceptable salts, hydrates or solvates of the 2-(substituted benzene miscellaneous group) aromatic formic acid, which can be used as an FTO target inhibitor for treating diseases related to the FTO target. The compound can be used for treating human myelodysplastic syndrome, such as leukemia, lymphoma, myelodysplastic syndrome, obesity, metablic syndrome (MS), type II diabetes (Type 2 diabetes, T2D), Alzheimer's disease, breast cancer, kidney cancer, colorectal cancer, pancreatic cancer, liver cancer, small cell lung cancer, human bone marrow rhabdomyoma, pancreatic cancer, malignant glioblast brain tumor and the like. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry. Specifically, the present invention discloses 2-(substituted benzyl) aromatic carboxylic acid and its derivative compounds as shown in the following general formula (I) and pharmaceutically acceptable salts, hydrates or solvates thereof, which can be used as inhibitors of FTO targets for treating diseases related to FTO targets, such as leukemia, lymphoma, myelodysplastic syndrome, obesity, metabolic syndrome (MS), type 2 diabetes (T2D), Alzheimer's disease, breast cancer, renal cancer, colorectal cancer, pancreatic cancer, liver cancer, small cell lung cancer, human bone marrow rhabdomyosarcoma, pancreatic cancer, malignant glioblastoma brain tumor, etc. Background Art

[0002] N6-methyladenosine 6 A) is the most abundant chemical modification on eukaryotic messenger RNA (mRNA). Its modification state is dynamically and reversibly regulated by methyltransferases and demethylases, affecting RNA splicing, translation, stability, and the epigenetics of certain non-coding RNAs, mediating a variety of physiological and pathological processes.

[0003] Where m 6 A demethylase fat mass and obesity-associated protein (FTO), in Fe 2+ With the assistance of α-ketoglutarate, FTO catalyzes the removal of m6A methylation modification, dynamically regulates the m6A modification level, and participates in the processes of glucose and lipid metabolism and nerve conduction defect repair. FTO has been shown to be associated with the occurrence and progression of obesity, type II diabetes, Alzheimer's disease, non-alcoholic fatty liver disease, and a variety of hematological tumors and solid tumors. In addition, abnormal expression of FTO protein in tumors often leads to m 6 Disordered A modification levels have a significant impact on the maintenance of tumor stem cells, the proliferation, migration and invasion of tumor cells, and even the sensitivity to radiotherapy and chemotherapy.

[0004] Therefore, the development of small molecule inhibitors targeting FTO has the potential to be used for the treatment of various diseases and there is still a huge demand. Summary of the invention

[0005] The purpose of the present invention is to provide an inhibitor that specifically targets FTO, and to treat diseases such as leukemia, lymphoma, myelodysplastic syndrome, obesity, metabolic syndrome (MS), type 2 diabetes (T2D), Alzheimer's disease, breast cancer, kidney cancer, colorectal cancer, pancreatic cancer, liver cancer, small cell lung cancer, human bone marrow rhabdomyosarcoma, pancreatic cancer, and malignant glioblastoma by inhibiting the enzymatic function of FTO or the signal transduction process mediated by FTO protein.

[0006] In the first aspect of the present invention, there is provided a compound represented by the following formula (I), and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof:

[0007]

[0008] in,

[0009] A1, A2 and A3 are each independently N, CH or CR3;

[0010] R1 is selected from the following group: C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 3-8 membered heterocyclyl, C3-C8 cycloalkyl-O-, 3-8 membered heterocyclyl-O-; and said R1 may be replaced by one or more R f Substitute, the R f Selected from the group consisting of deuterium, halogen, carbonyl (=O), carboxyl, hydroxyl, amino, nitro, cyano, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkoxycarbonyl, C1-C6 acylamino, C2-C6 ester, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C6~C 10 Aryl, substituted or unsubstituted five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, carbocyclic ring as described in 3-12-membered cycloalkyl;

[0011] R2 is selected from the following groups: H, halogen atoms, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C 3～ C 12 Cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 amide, carbonyl, carboxyl, hydroxyl, amino, nitro, cyano;

[0012] R3 is selected from the following group: H, halogen atoms, hydroxyl, amino, nitro, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 alkoxycarbonyl, substituted or unsubstituted C3-C8 cycloalkyl;

[0013] X has the following structure: COOH, CONH2, CONHOH, CONHR e 、CONHOR e 、CON(OH)R e 、COOR e ; R e is a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group, or a substituted or unsubstituted 3-8 membered heterocyclic group;

[0014] R a , R b Each is independently selected from the following group: halogen, -OH, CN, NO2, NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted -(CH2) m -C1~C6 alkoxy, substituted or unsubstituted -(CH2) m -C1~C6 alkylthio, substituted or unsubstituted -(CH2) m -3- to 8-membered heterocyclic group, substituted or unsubstituted -O-(CH2) m -3- to 8-membered heterocyclic group, substituted or unsubstituted C3- to C8-cycloalkyl group, substituted or unsubstituted 3- to 8-membered heterocyclic group, substituted or unsubstituted C3- to C8-cycloalkyloxy group, substituted or unsubstituted C6- to C8- 10 Aryl, substituted or unsubstituted 5-10 membered heteroaryl; wherein m is selected from the following group: 0, 1, 2, 3, 4, or 5;

[0015] R c , R d Each is independently selected from the following group: H, halogen, -OH, CN, NO2, NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclic group, substituted or unsubstituted C6-C 10 Aryl, substituted or unsubstituted 5- to 10-membered heteroaryl;

[0016] Ring A is a group selected from the following group: a naphthalene ring, a 3- to 10-membered heterocyclic group or a 5- to 10-membered heteroaryl group;

[0017] n is 0, 1, 2, 3 or 4;

[0018] The substitution mentioned herein refers to the substitution of one or more hydrogen atoms by a substituent selected from the group consisting of deuterium, a halogen atom, a carbonyl group (=O), a carboxyl group, a hydroxyl group, an amino group, a nitro group, a cyano group, a C1-C6 alkoxy group, a deuterated C1-C6 alkoxy group, a halogenated C1-C6 alkoxy group, a C1-C6 alkylamino group, a C1-C6 alkoxycarbonyl group, a C1-C6 acylamino group, a C2-C6 ester group, a C1-C6 alkyl group, a halogenated C1-C6 alkyl group, a C6-C 10 Aryl, five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, C3-C 12 Cycloalkyl;

[0019] The heterocycle or heterocyclic group may be a saturated or partially unsaturated structure, but not aromatic; the carbocycle, heterocycle, aromatic ring, heteroaromatic ring may be a monocyclic, spirocyclic, fused or bridged ring; the heterocyclic group or heteroaromatic group may each independently contain 1, 2, 3 or 4 heteroatoms selected from O, S and N.

[0020] In another preferred embodiment, the R1 is selected from the following group: C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 3-8 membered heterocyclic group, C3-C8 cycloalkyl-O-; and the R1 is optionally replaced by one or more R f Substitute, the R f Selected from the group consisting of deuterium, halogen, carbonyl (=O), carboxyl, hydroxyl, amino, nitro, cyano, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkoxycarbonyl, C1-C6 acylamino, C2-C6 ester, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C6~C 10 Aryl, substituted or unsubstituted five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, carbocyclic ring as described in 3-12-membered cycloalkyl;

[0021] R a , R b Each is independently selected from the following group: halogen, -OH, CN, NO2, NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted -(CH2) m -C1~C6 alkoxy, substituted or unsubstituted -(CH2) m -C1~C6 alkylthio, substituted or unsubstituted C3~C8 cycloalkyl, substituted or unsubstituted -(CH2) m -3- to 8-membered heterocyclic group, substituted or unsubstituted -O-(CH2) m -3- to 8-membered heterocyclic group, substituted or unsubstituted C3- to C8-cycloalkyloxy group, substituted or unsubstituted C6- to C 10 Aryl, substituted or unsubstituted 5-10 membered heteroaryl; wherein m is selected from the following group: 0, 1, or 2;

[0022] Rc , R d Each is independently selected from the following group: H, halogen, C1-C6 alkyl.

[0023] In another preferred embodiment, the R c and R d Each is independently H.

[0024] In another preferred embodiment, the X is selected from a substituted or unsubstituted structure selected from the following group:

[0025]

[0026] wherein n is 0, 1, 2, 3, 4, 5 or 6; R e R is selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclic group; g Selected from H, OH.

[0027] In another preferred embodiment, the A ring is selected from the following group: pyridine, pyrimidine, pyridazine, tetrazine, triazine, pyrrole, thiophene, furan, tetrazole, triazole, imidazole, thiazole, oxazole, pyrazole, isothiazole, isoxazole, oxadiazole, thiadiazole, morpholine, dihydropiperidine, thiomorpholine, piperidine, piperazine, tetrahydropyran, dihydropyran, pyrroline, tetrahydrothiophene, tetrahydrofuran, oxetane, thiazole, and azetidine.

[0028] In another preferred embodiment, the A ring is selected from the following group:

[0029]

[0030] In another preferred embodiment, the R a , R b Each is independently selected from the following group: F, Cl, Br, methyl, ethyl, cyclopropane, isopropyl, methoxy, methylthio, -CH2OCH3, trifluoromethyl, trifluoromethoxy, difluoromethoxy, deuterated methoxy.

[0031] In another preferred embodiment, the compound of formula (I) is selected from the following group:

[0032]

[0033]

[0034]

[0035]

[0036]

[0037] In the second aspect of the present invention, there is provided a use of the compound of formula (I) as described in the first aspect of the present invention and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof, which is used for a use selected from the group consisting of:

[0038] (a) preparing a drug for treating a disease associated with FTO protein activity or expression;

[0039] (b) preparing targeted inhibitors of FTO protein activity;

[0040] (c) non-therapeutic inhibition of FTO protein activity in vitro; and / or

[0041] (d) Treating diseases related to FTO activity or expression.

[0042] In another preferred embodiment, the disease is selected from the following group: leukemia, lymphoma, myelodysplastic syndrome, obesity, metabolic syndrome (MS), type 2 diabetes (T2D), Alzheimer's disease, breast cancer, kidney cancer, colorectal cancer, pancreatic cancer, liver cancer, small cell lung cancer, human bone marrow rhabdomyosarcoma, pancreatic cancer, and malignant glioblastoma.

[0043] In the third aspect of the present invention, a pharmaceutical composition is provided, comprising: (i) an effective amount of a compound of formula (I), and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof; and (ii) a pharmaceutically acceptable carrier.

[0044] In the fourth aspect of the present invention, a method for inhibiting the activity of FTO protein is provided, which comprises the steps of: administering to the inhibited object an inhibitory effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof as described in the first aspect of the present invention, or administering to the inhibited object an inhibitory effective amount of a pharmaceutical composition as described in the third aspect of the present invention.

[0045] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0046] After extensive and in-depth research, a large number of screening and testing, the inventors have provided a class of compounds as shown in formula (I), which can efficiently and selectively inhibit the activity of FTO protein, and its cell activity is significantly improved compared with existing FTO inhibitors. Based on the above findings, the inventors have completed the present invention.

[0047] the term

[0048] The term "C1-C6 alkyl" refers to a straight or branched chain alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like.

[0049] The term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, or the like.

[0050] The term C3-C8 cycloalkyl refers to a saturated or unsaturated cyclic group having 3 to 8 carbon atoms, the ring skeleton of which is composed entirely of carbon atoms. In some preferred embodiments, the carbon ring may be saturated or partially unsaturated, but not aromatic, such as cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane or similar structures.

[0051] The term "C6~C 10 The term "aryl" refers to an aromatic group having 6, 7, 8, 9 or 10 carbon atoms, including monocyclic or bicyclic aromatic groups, such as phenyl, naphthyl, or the like.

[0052] The term "3- to 10-membered heterocyclic group" refers to a saturated or unsaturated (including aromatic) ring system substituent having 3, 4, 5, 6, 7, 8, 9 or 10 members and having one or more heteroatoms selected from O, S, N or P, such as tetrahydrofuran, pyrroline, oxetane, or similar groups, preferably a 4- to 9-membered heterocyclic group.

[0053] The term "5- to 6-membered heteroaryl" refers to a 5- or 6-membered ring system having one or more heteroatoms selected from O, S, N or P, and having aromatic ring system substituents, such as pyridyl, thienyl, and furanyl. The term "5- to 10-membered heteroaryl" has a similar definition, and refers to a 5-, 6-, 7-, 8-, 9-, or 10-membered ring system having one or more heteroatoms selected from O, S, N or P, wherein the heteroaryl group as a whole forms an aromatic structure.

[0054] Unless otherwise specified, in the present invention, the term "substituted" includes various isotope substitutions of hydrogen, such as deuteration.

[0055] The term "halogen" refers to F, Cl, Br and I.

[0056] In the present invention, the terms "comprising", "including" or "including" indicate that various components can be used together in the mixture or composition of the present invention. Therefore, the terms "consisting mainly of" and "consisting of" are included in the term "comprising".

[0057] In the present invention, the term "pharmaceutically acceptable" ingredient refers to a substance that is suitable for use in humans and / or animals without excessive adverse side effects (such as toxicity, irritation and allergic reaction), that is, has a reasonable benefit / risk ratio.

[0058] In the present invention, the term "effective amount" refers to the amount of a therapeutic agent to treat, alleviate or prevent a target disease or condition, or the amount that exhibits a detectable therapeutic or preventive effect. The exact effective amount for a subject depends on the subject's size and health, the nature and extent of the condition, and the therapeutic agent and / or combination of therapeutic agents selected for administration. Therefore, it is useless to specify an exact effective amount in advance. However, for a given condition, the effective amount can be determined by routine experiments, which a clinician is able to judge.

[0059] Unless otherwise specified, in the present invention, all compounds appearing are intended to include all possible optical isomers, such as single chiral compounds, or mixtures of various chiral compounds (i.e., racemates). Among all compounds of the present invention, each chiral carbon atom can be optionally in R configuration or S configuration, or a mixture of R configuration and S configuration.

[0060] As used herein, the term "compound of the present invention" refers to a compound of formula (I). The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compound of formula (I).

[0061] Compound of formula (I)

[0062]

[0063] in,

[0064] A1, A2 and A3 are each independently N, CH or CR3;

[0065] R1 is selected from the following group: C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 3-8 membered heterocyclyl, C3-C8 cycloalkyl-O-, 3-8 membered heterocyclyl-O-; and said R1 may be replaced by one or more R f Substitute, the R f Selected from the group consisting of deuterium, halogen, carbonyl (=O), carboxyl, hydroxyl, amino, nitro, cyano, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkoxycarbonyl, C1-C6 acylamino, C2-C6 ester, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C6~C 10 Aryl, substituted or unsubstituted five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, carbocyclic ring as described in 3-12-membered cycloalkyl;

[0066] R2 is selected from the following groups: H, halogen atoms, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C 3～ C 12 Cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 amide, carbonyl, carboxyl, hydroxyl, amino, nitro, cyano;

[0067] R3 is selected from the following group: H, halogen atoms, hydroxyl, amino, nitro, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 alkoxycarbonyl, substituted or unsubstituted C3-C6 cycloalkyl;

[0068] X has the following structure: COOH, CONH2, CONHOH, CONHR e 、CONHOR e 、CON(OH)R e 、COOR e ; R e is a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group, or a substituted or unsubstituted 3-8 membered heterocyclic group;

[0069] R a , R b Each is independently selected from the following group: halogen, -OH, CN, NO2, NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted -(CH2) m -C1~C6 alkoxy, substituted or unsubstituted -(CH2) m -C1~C6 alkylthio, substituted or unsubstituted -(CH2) m -3- to 8-membered heterocyclic group, substituted or unsubstituted -O-(CH2) m -3- to 8-membered heterocyclic group, substituted or unsubstituted C3- to C8-cycloalkyl group, substituted or unsubstituted C3- to C8-cycloalkoxy group, substituted or unsubstituted C6- to C8-cycloalkyl group 10 Aryl, substituted or unsubstituted 5-10 membered heteroaryl; wherein m is selected from the following group: 0, 1, 2, 3, 4, or 5;

[0070] R c , R d Each is independently selected from the following group: H, halogen, -OH, CN, NO2, NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclic group, substituted or unsubstituted C6-C 10Aryl, substituted or unsubstituted 5- to 10-membered heteroaryl;

[0071] Ring A is a group selected from the group consisting of a naphthalene ring, a 3- to 10-membered heterocyclic group, or a 5- to 10-membered heteroaryl group;

[0072] n is 0, 1, 2, 3 or 4;

[0073] The substitution mentioned herein refers to the substitution of one or more hydrogen atoms by a substituent selected from the group consisting of deuterium, a halogen atom, a carbonyl group (=O), a carboxyl group, a hydroxyl group, an amino group, a nitro group, a cyano group, a C1-C6 alkoxy group, a deuterated C1-C6 alkoxy group, a halogenated C1-C6 alkoxy group, a C1-C6 alkylamino group, a C1-C6 alkoxycarbonyl group, a C1-C6 acylamino group, a C2-C6 ester group, a C1-C6 alkyl group, a halogenated C1-C6 alkyl group, a C6-C 10 Aryl, five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, C3-C 12 Cycloalkyl;

[0074] The heterocycle or heterocyclic group may be a saturated or partially unsaturated structure, but not aromatic; the carbocycle, heterocycle, aromatic ring, heteroaromatic ring may be a monocyclic, spirocyclic, fused or bridged ring; the heterocyclic group or heteroaromatic group may each independently contain 1, 2, 3 or 4 heteroatoms selected from O, S and N.

[0075] The pharmaceutically acceptable salt is selected from: inorganic acid salts, organic acid salts, inorganic alkali salts, or organic alkali salts. Preferably, it is an inorganic acid salt or an organic acid salt, and the inorganic acid salt is selected from hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate, and acid phosphate; the organic acid salt is selected from formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, salicylate, picrate, glutamate, salicylate, ascorbate, camphorate, and camphorsulfonate.

[0076] In a preferred embodiment, the acceptable salt is an inorganic base or an organic base salt, and the inorganic base is selected from aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganic salts, manganous salts, potassium salts, sodium salts, zinc salts, etc. Ammonium salts, calcium salts, magnesium salts, potassium salts and sodium salts are particularly preferred. Salts derived from pharmaceutically acceptable organic non-toxic bases, including salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydroxocobalamin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, guazine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like.

[0077] Pharmaceutical compositions and methods of administration

[0078] Since the compounds of the present invention have excellent inhibitory activity against FTO protein, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredients can be used to treat, prevent and alleviate diseases related to FTO activity or expression. According to the prior art, the compounds of the present invention can be used to treat the following diseases: leukemia, lymphoma, myelodysplastic syndrome, obesity, metabolic syndrome (metabolic syndrome, MS), type II diabetes (Type 2diabetes, T2D), Alzheimer's disease, breast cancer, renal cancer, colorectal cancer, pancreatic cancer, liver cancer, small cell lung cancer, human bone marrow rhabdomyosarcoma, pancreatic cancer, malignant glioblastoma brain tumor and other diseases.

[0079] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1 to 2000 mg of the compound of the present invention per dose, and more preferably, contains 5 to 200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

[0080] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0081] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0082] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0083] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0084] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0085] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0086] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0087] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0088] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0089] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0090] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 5 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill range of a skilled physician.

[0091] The main advantages of the present invention include:

[0092] (1) The present invention provides a class of demethylase FTO inhibitors, which can be used at very low concentrations (IC 50 The activity of FTO protein was inhibited at 40 μM (the value was usually lower than 30 μM).

[0093] (2) The FTO inhibitor of the present invention has high selectivity and has the function of regulating m6 The function of A.

[0094] (3) The FTO inhibitor of the present invention has strong anti-cell proliferation activity.

[0095] (4) The FTO protein activity inhibitor of the present invention exhibits good pharmacokinetic effects in mice.

[0096] The present invention will be further described below in conjunction with specific implementation. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0097] The experimental reagents used in the experiment were mainly purchased from China National Pharmaceutical Group, Sigma and Shanghai Biotechnology.

[0098] Example 1: Synthesis of R02

[0099]

[0100] Step 1: In a 250 mL round-bottom flask, add A-1a (4.2 g, 15 mmol, 1.0 eq), K2CO3 (2.1 g, 15 mmol, 1.0 eq), diethyl sulfate (2.3 g, 15 mmol, 1.0 eq), N,N-dimethylformamide (75 mL) in sequence, and stir at room temperature overnight. Add water (75 mL) to the system, adjust the pH to neutral with saturated ammonium chloride solution, add ethyl acetate (75 mL × 3) to the solution, combine the organic phases, wash the organic phases with water (75 mL × 3), wash with saturated brine (75 mL × 3), dry the organic phase with anhydrous magnesium sulfate, and filter. The filtrate is concentrated, and the crude product is separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-2a (yellow oil, 4.2 g), which is directly used in the next step.

[0101] Step 2: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 2,6-dichloro-4-bromoaniline (2.8 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3a (white solid, 3.2 g), which was directly used in the next step.

[0102] Step 3: In a 25 mL round-bottom flask, compound A-3a (419 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL) were added in sequence, heated to 100 ° C and stirred overnight in a nitrogen atmosphere. The reaction solution was concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound A-4a-1 (white solid, 200 mg), which was directly used in the next step.

[0103] Step 4: Weigh compound A-4a-1 (200 mg, 0.49 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3.2 mL / 6.4 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.6 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), concentrated to dryness under reduced pressure, and the crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R02 (white solid, 97 mg). LCMS: [M+H] + =407. 1 H NMR (500MHz, DMSO-d6) δ8.51(s,1H),7.46–7.41(m,3H),7.35(d,J=2.6Hz,1H),6.90(dd,J=8.4,2.6Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0104] Example 2: Synthesis of R29

[0105]

[0106] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2-fluoro-6-methoxyaniline (2.6 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3p (yellow solid, 2.6 g), which was directly used in the next step.

[0107] Step 2: In a 25 mL round-bottom flask, add compound A-3p (398 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound A-4p-1 (yellow solid, 254 mg), which is directly used in the next step.

[0108] Step 3: Weigh the compound A-4p-1 (207 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), concentrated to dryness under reduced pressure, and the crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R29 (yellow solid, 119 mg).

[0109] LCMS: [M+H] + =387; 1 H-NMR (500MHz, DMSO-d6) δ10.75 (s, 1H), 7.45 (d, J = 3.2

[0110] Hz,1H),6.90(dd,J=11.3,1.8Hz,1H),6.86–6.83(m,1H),6.70(dd,J=8.8,3.2Hz,1H ),6.29(dd,J=8.8,4.9Hz,1H),3.82(s,3H),3.66(s,3H),2.45(s,3H),2.28(s,3H).

[0111] Example 3: Synthesis of R35

[0112]

[0113] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 2,6-difluoro-4-bromoaniline (2.4 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3c (white solid, 3.4 g), which was directly used in the next step.

[0114] Step 2: In a 25 mL round-bottom flask, add compound A-3c (402 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound A-4c-1 (white solid, 183 mg), which is directly used in the next step.

[0115] Step 3: Weigh the compound A-4c-1 (156 mg, 0.38 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (44 mg, 1.85 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R35 (white solid, 88 mg). LCMS: [M+H] + =375;1 H-NMR (400MHz, DMSO-d6) δ9.04 (s, 1H), 7.41 (d, J = 3.1Hz, 1H), 7.36–7.24 (m, 2H), 7.09 (d d,J=9.1,3.1Hz,1H),6.60(dt,J=9.1,2.7Hz,1H),3.73(s,3H),2.46(s,3H),2.28(s,3H).

[0116] Example 4: Synthesis of R36

[0117]

[0118] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 2,6-dimethyl-4-bromoaniline (2.3 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3d (white solid, 3.2 g), which was directly used in the next step.

[0119] Step 2: In a 25 mL round-bottom flask, add compound A-3d (402 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound A-4d-1 (white solid, 174 mg), which is directly used in the next step.

[0120] Step 3: Weigh the compound A-4d-1 (156 mg, 0.38 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (44 mg, 1.85 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R36 (white solid, 94 mg). LCMS: [M+H]+ =367; 1 H-NMR (400MHz, DMSO-d6) δ8.85 (s, 1H), 7.39 (d, J = 3.1Hz, 1H), 7.17 (s, 2H), 6.98 (dd, J = 9. 1,3.1Hz,1H),6.13(d,J=9.0Hz,1H),3.69(s,3H),2.42(s,3H),2.25(s,3H),2.16(s,6H).

[0121] Example 5: Synthesis of R37

[0122]

[0123] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2-chloro-6-methylaniline (2.6 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1) to obtain compound A-3e (white solid, 3.4 g), which was directly used in the next step.

[0124] Step 2: In a 25 mL round-bottom flask, add compound A-3e (402 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound A-4e-1 (white solid, 174 mg), which is directly used in the next step.

[0125] Step 3: Weigh the compound A-4e-1 (156 mg, 0.38 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (44 mg, 1.85 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R37 (white solid, 120 mg). LCMS: [M+H] + =387; 1 H-NMR (400MHz, DMSO-d6) δ9.03(s,1H),7.46(d,J=2.0Hz,1H),7.41(d,J=3.1Hz,1H),7.34(d,J=2.0Hz,1H ),7.01(dd,J=9.0,3.1Hz,1H),6.21(d,J=9.0Hz,1H),3.70(s,3H),2.44(s,3H),2.26(s,3H),2.21(s,3H).

[0126] Example 6: Synthesis of R38

[0127]

[0128] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2-fluoro-6-methylaniline (2.4 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1) to obtain compound A-3f (white solid, 3.4 g), which was directly used in the next step.

[0129] Step 2: In a 25 mL round-bottom flask, add compound A-3f (402 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound A-4f-1 (white solid, 174 mg), which is directly used in the next step.

[0130] Step 3: Weigh the compound A-4f-1 (156 mg, 0.38 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (44 mg, 1.85 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R38 (white solid, 134 mg). LCMS: [M+H] + =371; 1 H-NMR (400MHz, DMSO-d6) δ8.93 (s, 1H), 7.40 (d, J = 3.1Hz, 1H), 7.26–7.17 (m, 2H), 7.04 (dd, J = 9. 1,3.1Hz,1H),6.37(dd,J=9.1,3.0Hz,1H),3.71(s,3H),2.44(s,3H),2.27(s,3H),2.24(s,3H).

[0131] Example 7: Synthesis of R39

[0132]

[0133] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 2-chloro-6-fluoro-4-bromoaniline (2.6 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3b (white solid, 3.2 g), which was directly used in the next step.

[0134] Step 2: In a 25 mL round-bottom flask, add compound A-3b (402 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound A-4b-1 (white solid, 156 mg), which is directly used in the next step.

[0135] Step 3: Weigh the compound A-4b-1 (156 mg, 0.38 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (44 mg, 1.85 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R39 (white solid, 90 mg). LCMS: [M+H] + =391; 1 H-NMR (400MHz, DMSO-d6) δ8.84(s,1H),7.63(dd,J=10.3,2.4Hz,1H),7.34(dd,J=2.5,1.3Hz,1H),7.28(d,J= 3.1Hz, 1H), 6.96 (dd, J=9.1, 3.1Hz, 1H), 6.39 (dd, J=9.1, 3.4Hz, 1H), 3.68 (s, 3H), 2.22 (s, 3H), 2.00 (s, 3H).

[0136] Example 8: Synthesis of R40

[0137]

[0138] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2-chloro-6-(trifluoromethyl)aniline (3.2 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1) to obtain compound A-3g (white solid, 3.4 g), which was directly used in the next step.

[0139] Step 2: In a 25 mL round-bottom flask, add compound A-3g (402 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound A-4g-1 (white solid, 175 mg), which is directly used in the next step.

[0140] Step 3: Weigh the compound A-4g-1 (156 mg, 0.38 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (44 mg, 1.85 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R40 (white solid, 98 mg). LCMS: [M+H] + =441; 1 H-NMR (400MHz, DMSO-d6) δ9.36(s,1H),7.96(d,J=2.0Hz,1H),7.78(d,J=2.0Hz,1H),7.41(d,J=3.1 Hz,1H),7.03(dd,J=9.1,3.1Hz,1H),6.29(d,J=9.0Hz,1H),3.72(s,3H),2.46(s,3H),2.28(s,3H).

[0141] Example 9: Synthesis of R41

[0142]

[0143] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2,6-diisopropylaniline (3.0 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3h (white solid, 3.1 g), which was directly used in the next step.

[0144] Step 2: In a 25 mL round-bottom flask, add compound A-3h (434 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound A-4h-1 (off-white solid 151 mg), which is directly used in the next step.

[0145] Step 3: Weigh the compound A-4h-1 (151 mg, 0.33 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (39.6 mg, 1.65 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R41 (white solid, 93 mg). LCMS: [M+H] + =423; 1H-NMR (400MHz, DMSO-d6) δ8.88(s,1H),7.38(d,J=3.1Hz,1H),7.23(s,2H),6.99(dd,J=9.1,3.1Hz,1H),6.11(d,J=9. 1Hz,1H),3.68(s,3H),3.05(p,J=6.9Hz,2H),2.46(s,3H),2.29(s,3H),1.14(d,J=6.8Hz,6H),1.10(d,J=6.9Hz,6H).

[0146] Example 10: Synthesis of R44

[0147]

[0148] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2,6-dimethoxyaniline (2.7 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3i (white solid, 3.1 g), which was directly used in the next step.

[0149] Step 2: In a 25 mL round-bottom flask, add compound A-3i (410 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound A-4i-1 (white solid, 190 mg), which is directly used in the next step.

[0150] Step 3: Weigh the compound A-4i-1 (151 mg, 0.44 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (52.8 mg, 2.2 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R44 (yellow solid, 92 mg). LCMS: [M+H] + =399; 1 H-NMR(500MHz,DMSO-d6)δ8.72(s,1H),7.33(d,J=3.1Hz,1H),6.97(dd,J=9.1,3.1Hz,1H) ,6.72(s,2H),6.30(d,J=9.2Hz,1H),3.78(s,6H),3.69(s,3H),2.47(s,3H),2.30(s,3H).

[0151] Example 11: Synthesis of R114

[0152]

[0153] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2-ethyl-6-methyl-aniline (2.5 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3n (white solid, 2.3 g), which was directly used in the next step.

[0154] Step 2: In a 25 mL round-bottom flask, add compound A-3n (392 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound A-4n-1 (white solid, 208 mg), which is directly used in the next step.

[0155] Step 3: Weigh the compound A-4n-1 (208 mg, 0.51 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (62.4 mg, 2.6 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R114 (white solid, 87 mg). LCMS: [M+H] + =381; 1 H-NMR(500MHz,DMSO-d6)δ8.89(s,1H),7.39(d,J=3.1Hz,1H),7.22–7.13(m,2H),6.98(dd,J=9.1,3.1Hz,1H),6.12 (d,J=9.1Hz,1H),3.69(s,4H),2.53(d,J=7.5Hz,2H),2.43(s,3H),2.26(s,3H),2.14(s,3H),1.09(t,J=7.5Hz,3H).

[0156] Example 12: Synthesis of R116

[0157]

[0158] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2-isopropyl-6-methyl-aniline (2.7 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3o (white solid, 1.9 g), which was directly used in the next step.

[0159] Step 2: In a 25 mL round-bottom flask, add compound A-3o (406 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound A-4o-1 (brown solid, 169 mg), which is directly used in the next step.

[0160] Step 3: Weigh the compound A-4o-1 (169 mg, 0.40 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R116 (yellow solid, 79 mg). LCMS: [M+H] + =395; 1 H-NMR(500MHz,DMSO-d6)δ8.91(s,1H),7.38(d,J=3.1Hz,1H),7.26–7.14(m,2H),6.98(dd,J=9.1,3.1Hz,1H),6.11(d, J=9.1Hz,1H),3.69(s,3H),3.09(p,J=6.9Hz,1H),2.44(s,3H),2.27(s,3H),2.13(s,3H),1.13(dd,J=22.0,6.9Hz,7H).

[0161] Example 13: Synthesis of R27

[0162]

[0163] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2,6-dicyclopropylaniline (2.9 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3q (yellow solid, 2.5 g), which was directly used in the next step.

[0164] Step 2: In a 25 mL round-bottom flask, add compound A-3q (430 mg, 1.0 mmol, 1.0 eq), 3,5-dimethyl-1H-pyrazole-4-boronic acid (210 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound A-4q-2 (yellow solid, 228 mg), which is directly used in the next step.

[0165] Step 3: Weigh the compound A-4q-2 (223 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R27 (yellow solid, 112 mg). LCMS: [M+H] + =418; 1 H-NMR (500MHz, DMSO-d6) δ9.15(s,1H),7.38(d,J=3.1Hz,1H),7.01(dd,J=9.1,3.1Hz,1H),6.67(s,2H),6.35(d,J= 9.1Hz,1H),3.69(s,4H),2.26(s,6H),1.88(tt,J=8.5,5.3Hz,2H),0.83(d,J=34.7Hz,4H),0.63(d,J=37.8Hz,4H).

[0166] Example 14: Synthesis of R32

[0167]

[0168] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2-ethyl-6-methylaniline (2.5 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3n (yellow solid, 3.1 g), which was directly used in the next step.

[0169] Step 2: In a 25 mL round-bottom flask, add compound A-3n (392 mg, 1.0 mmol, 1.0 eq), 3,5-dimethyl-1H-pyrazole-4-boronic acid (210 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound A-4n-2 (yellow solid, 278 mg), which is directly used in the next step.

[0170] Step 3: Weigh the compound A-4n-2 (204 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R32 (yellow solid, 145 mg). LCMS: [M+H] + =380; 1 H-NMR (400MHz, DMSO-d6) δ8.89 (s, 1H), 7.38 (d, J = 3.1Hz, 1H), 7.07 (q, J = 2.2Hz, 2H), 6.97 (dd, J = 9. 1,3.1Hz,1H),6.12(d,J=9.1Hz,1H),3.68(s,3H),2.22(s,6H),2.12(s,3H),1.08(t,J=7.5Hz,3H).

[0171] Example 15: Synthesis of R33

[0172]

[0173] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2-cyclopropyl-6-fluoroaniline (2.7 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3k (yellow solid, 2.5 g), which was directly used in the next step.

[0174] Step 2: In a 25 mL round-bottom flask, add compound A-3k (408 mg, 1.0 mmol, 1.0 eq), 3,5-dimethyl-1H-pyrazole-4-boronic acid (210 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound A-4k-2 (brown solid, 264 mg), which is directly used in the next step.

[0175] Step 3: Weigh the compound A-4k-2 (212 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R33 (brown solid, 101 mg). LCMS: [M+H] + =396; 1H-NMR (400MHz, DMSO-d6) δ9.34–8.97(m,1H),7.39(s,1H),7.03(td,J=7.2,2.9Hz,2H),6.67(d,J=1.8Hz, 1H), 6.45 (d, J = 9.2Hz, 1H), 3.70 (s, 4H), 2.21 (s, 7H), 1.93 (s, 1H), 1.01–0.85 (m, 2H), 0.77–0.59 (m, 2H).

[0176] Example 16: Synthesis of R55

[0177]

[0178] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2-chloro-6-methoxyaniline (2.8 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3j (white solid, 3.1 g), which was directly used in the next step.

[0179] Step 2: In a 25 mL round-bottom flask, add compound A-3j (410 mg, 1.0 mmol, 1.0 eq), 3,5-dimethyl-1H-pyrazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 6:1) to obtain compound A-4j-2 (brown solid 190 mg), which is directly used in the next step.

[0180] Step 3: Weigh the compound A-4j-2 (190 mg, 0.44 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (52.8 mg, 2.2 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R55 (yellow solid, 132 mg). LCMS: [M+H] + =402; 1 H-NMR(500MHz,DMSO-d6)δ8.93(s,1H),7.37(d,J=3.1Hz,1H),7.10(dd,J=9.8,5.4Hz,1H),7.01( dd,J=9.0,3.2Hz,2H),6.31(dd,J=9.2,1.1Hz,1H),3.79(s,3H),3.70(s,3H),2.41–2.21(m,6H).

[0181] Example 17: Synthesis of R96

[0182]

[0183] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2,6-ethylaniline (2.7 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3m (white solid, 2.4 g), which was directly used in the next step.

[0184] Step 2: In a 25 mL round-bottom flask, add compound A-3m (406 mg, 1.0 mmol, 1.0 eq), 3,5-dimethyl-1H-pyrazole-4-boronic acid (210 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound A-4m-2 (yellow solid, 197 mg), which is directly used in the next step.

[0185] Step 3: Weigh the compound A-4m-2 (197 mg, 0.47 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.4 mL / 4.8 mL), add lithium hydroxide (57.6 mg, 2.4 mmol, 5.0 eq) in water (1.2 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R96 (yellow solid, 75 mg). LCMS: [M+H] + =394; 1 H-NMR (500MHz, DMSO-d6) δ8.87(s,1H),7.37(d,J=3.1Hz,1H),7.09(s,2H),6.97(dd,J=9.1,3.2H z,1H),6.13(d,J=9.1Hz,1H),3.68(s,3H),2.49–2.44(m,3H),2.23(s,6H),1.08(t,J=7.6Hz,6H).

[0186] Example 18: Synthesis of R31

[0187]

[0188] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-3-chloro-2,6-dimethylaniline (2.7 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3r (yellow solid, 2.9 g), which was directly used in the next step.

[0189] Step 2: In a 25 mL round-bottom flask, add compound A-3r (413 mg, 1.0 mmol, 1.0 eq), pyridine-4-boric acid (185 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound A-4r-4 (yellow solid, 267 mg), which is directly used in the next step.

[0190] Step 3: Weigh the compound A-4r-4 (206 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R31 (yellow solid, 122 mg). LCMS: [M+H] + =383; 1 H-NMR(500MHz,DMSO-d6)δ8.69–8.61(m,2H),7.50(dt,J=5.3,2.9Hz,1H),7.49–7.43(m,2H) ,7.22(s,1H),6.75–6.66(m,1H),6.01–5.90(m,1H),3.66(s,4H),2.25(s,3H),2.17(s,3H).

[0191] Example 19: Synthesis of R61

[0192]

[0193] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, A-2a (4.2 g, 14 mmol, 1.2 eq), 4-bromo-2,6-dichloroaniline (2.8 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound A-3a (yellow solid, 2.9 g), which was directly used in the next step.

[0194] Step 2: In a 25 mL round-bottom flask, add compound A-3a (419 mg, 1.0 mmol, 1.0 eq), 2-ethoxypyridine-4-boric acid (251 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound A-4a-5 (brown solid, 268 mg), which is directly used in the next step.

[0195] Step 3: Weigh the compound A-4a-5 (231 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R61 (brown solid, 123 mg). LCMS: [M+H] + =433; 1 H-NMR(500MHz,DMSO-d6)δ9.26(s,1H),8.23(d,J=5.3Hz,1H),8.05(s,2H),7.44–7.38(m,2H),7.23(d,J=1.6Hz,1 H),7.03(dd,J=9.0,3.1Hz,1H),6.32(d,J=9.0Hz,1H),4.36(q,J=7.0Hz,2H),3.72(s,3H),1.35(t,J=7.0Hz,3H).

[0196] Example 20: Synthesis of R82

[0197]

[0198] Step 1: In a 25 mL round-bottom flask, add compound A-3a (419 mg, 1.0 mmol, 1.0 eq), 2,3-dichloropyridine-4-boric acid (288 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL), heat to 100 ° C and stir overnight in a nitrogen atmosphere. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound A-4a-6 (brown solid 278 mg), which is directly used in the next step.

[0199] Step 2: Weigh the compound A-4a-6 (243 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R82 (brown solid 113 mg). LCMS: [M+H] + =459; 1 H-NMR(500MHz,DMSO-d6)δ9.75(s,1H),8.46(d,J=4.9Hz,1H),8.17(s,1H),7.76(s,2H),7.61(d,J=4.9Hz ,1H),7.56(s,1H),7.31(d,J=2.9Hz,1H),6.95(dd,J=9.0,2.9Hz,1H),6.33(d,J=8.9Hz,1H),3.74(s,3H).

[0200] Example 21: Synthesis of R22

[0201]

[0202] Step 1: In a 25 mL round-bottom flask, compound A-3a (210 mg, 0.5 mmol, 1.0 eq), 2-methoxypyridine-4-boronic acid (92 mg, 0.6 mmol, 1.2 eq), Pd(dppf)Cl2 (37 mg, 0.05 mmol, 0.1 eq), K2CO3 (138 mg, 1.0 mmol, 2.0 eq), dioxane / water (2.5 mL / 0.25 mL, 10:1, v / v) were weighed and heated to 100 ° C in a nitrogen atmosphere and stirred overnight. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound A-4a-7 (white solid, 179 mg), which was used directly in the next step.

[0203] Step 2: Weigh the compound A-4a-7 (179 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R22 (yellow solid, 145 mg). LCMS: [M+H] + =419. 1 H-NMR(500MHz,Chloroform-d)δ8.55(s,1H),8.29(d,J=4.7Hz,1H),7.69(s,2H),7.43(d ,J=8.4Hz,1H),7.38–7.33(m,2H),6.90(dt,J=5.2,2.7Hz,2H),3.95(s,3H),3.80(s,3H).

[0204] Example 22: Synthesis of R50

[0205]

[0206] In a 50mL round-bottom flask, compound R39 (124mg, 0.3mmol, 1.0eq), NH2O-THP (18mg, 0.15mmol, 0.5eq), HOBT (49mg, 0.36mmol, 1.2eq), EDCI (69mg, 0.36mmol, 1.2eq), N,N-dimethylformamide (6mL) were weighed and stirred at room temperature overnight. The reaction solution was added with ice to precipitate solids, which were filtered, washed with water (2mL×3), washed with n-hexane (2mL×3), and dried to obtain the compound (white solid, 122mg), which was directly used in the next step. In a 50mL round-bottom flask, the product of the previous step (122mg, 0.25mmol, 1.0eq), trifluoroacetic acid (570mg, 5mmol, 20.0eq), dichloromethane / methanol (1.3mL / 1.3mL) were weighed and stirred at room temperature overnight. The reaction solution was directly filtered to obtain the target compound R50 (white solid, 37mg). LCMS: [M+H] + =406. 1 H-NMR (400MHz, DMSO-d6) δ9.94–9.88(m,2H),8.84(d,J=3.8Hz,1H),7.51(d,J=8.5Hz,1H),7.39(d,J=2.7Hz,1H),7.3 4(d,J=2.2Hz,1H),7.15(dd,J=12.2,2.1Hz,1H),6.91(dd,J=8.4,2.7Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0207] Example 23: Synthesis of R62

[0208]

[0209] Step 1: In a 500 mL round-bottom flask, under nitrogen atmosphere, B-1a (11.5 g, 50 mmol, 1.0 eq), cyclopropylboronic acid (6.5 g, 75 mmol, 1.5 eq), Pd(OAc)2 (1.1 g, 5 mmol, 0.1 eq), tricyclohexylphosphine (2.1 g, 7.5 mmol, 0.15 eq), K3PO4 (15.9 g, 75 mmol, 1.5 eq), toluene / water (250 mL / 25 mL) were added in sequence, and the reaction system was heated to 110 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound B-2a (yellow oil, 7.6 g), which was directly used in the next step.

[0210] Step 2: In a 500mL round-bottom flask, weigh B-2a (7.6g, 40mmol, 4.0eq), add water (56mL), concentrated hydrochloric acid (12M, 56mL), and cool in an ice-water bath. Slowly add sodium nitrite (0.8g, 12mmol, 1.2eq) aqueous solution (8mL) through a constant pressure dropping funnel. Continue to keep stirring at 0℃ for 30min after adding. Continue to slowly add potassium iodide (5.0g, 30mmol, 3.0eq) aqueous solution (20mL) through a constant pressure dropping funnel. After adding, heat to 90℃ and stir for 90min. After the reaction is completed, wash the solution with ethyl acetate (200mL), combine the organic phases, wash the organic phases with saturated sodium bisulfite solution (200mL), dry with anhydrous sodium sulfate, and filter. The filtrate is concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain the target B-3a (pink oil, 5.7g), which is directly used in the next step.

[0211] Step 3: In a 50 mL round-bottom flask, under nitrogen atmosphere, add B-3a (1.51 g, 5 mmol, 1.0 eq), 2,6-dichloro-4-bromoaniline (1.4 g, 6 mmol, 1.2 eq), Pd(OAc)2 (112 mg, 0.5 mmol, 0.1 eq), XantPhos (396 mg, 0.75 mmol, 0.15 eq), Cs2CO3 (2.44 g, 7.5 mmol, 1.5 eq), and toluene (50 mL) in sequence, heat the reaction system to 110 ° C, and react for 24 h. Concentrate by rotary evaporation and separate by silica gel column chromatography (100% petroleum ether) to obtain compound B-4a (yellow solid, 2.2 g), which is directly used in the next step.

[0212] Step 4: In a 25 mL round-bottom flask, compound B-4a (208 mg, 0.5 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (85 mg, 0.6 mmol, 1.2 eq), Pd(dppf)Cl2 (37 mg, 0.05 mmol, 0.1 eq), K2CO3 (138 mg, 1.0 mmol, 2.0 eq), dioxane / water (2.5 mL / 0.25 mL, 10:1, v / v) were weighed and heated to 100 ° C in a nitrogen atmosphere and stirred overnight. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound B-5a-1 (white solid, 184 mg), which was directly used in the next step.

[0213] Step 5: Weigh the compound B-5a-1 (172 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R62 (white solid, 78 mg). LCMS: [M+H] + =417; 1 H-NMR(500MHz,DMSO-d6)δ9.39(s,1H),7.65(s,2H),7.64(d,J=2.3Hz,1H),7.09(dd,J=8.6,2.3Hz,1H),6 .26(d,J=8.6Hz,1H),2.45(s,3H),2.27(s,3H),1.93–1.81(m,1H),0.93–0.81(m,2H),0.68–0.51(m,2H).

[0214] Example 24: Synthesis of R63

[0215]

[0216] In a 50mL round-bottom flask, compound R62 (125mg, 0.3mmol, 1.0eq), NH2O-THP (18mg, 0.15mmol, 0.5eq), HOBT (49mg, 0.36mmol, 1.2eq), EDCI (69mg, 0.36mmol, 1.2eq), N,N-dimethylformamide (6mL) were weighed and stirred at room temperature overnight. The reaction solution was added with ice to precipitate solids, which were filtered, washed with water (2mL×3), washed with n-hexane (2mL×3), and dried to obtain the compound (white solid, 120mg), which was directly used in the next step. In a 50mL round-bottom flask, the product of the previous step (120mg, 0.23mmol, 1.0eq), trifluoroacetic acid (524mg, 4.6mmol, 20.0eq), dichloromethane / methanol (1.3mL / 1.3mL) were weighed and stirred at room temperature overnight. The reaction solution was directly filtered to obtain the target compound R63 (white solid, 45 mg). LCMS: [M+H] + =432. 1H-NMR (400MHz, DMSO-d6) δ9.87(d,J=3.5Hz,1H),9.16(s,1H),7.80(dd,J=2.2,0.7Hz,1H),7.45–7.40(m,3H),7.24–7 .15(m,1H),2.72–2.65(m,1H),2.47(s,3H),2.40(s,3H),1.65(dd,J=10.7,5.6Hz,2H),1.40(dd,J=10.7,5.6Hz,2H).

[0217] Example 25: Synthesis of R71

[0218]

[0219] Step 1: In a 100 mL round-bottom flask, under nitrogen protection, compound 2c (3.1 g, 15 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.5 g, 18 mmol, 1.2 eq), Pd(PPh3)4 (3.5 g, 1.5 mmol, 0.1 eq), K3PO4 (8.0 g, 30 mmol, 2.0 eq), toluene / water (40 mL / 20 mL) were added successively, and the reaction system was heated to 80 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound M04 (white solid, 2.3 g).

[0220] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (1.5 g, 5 mmol, 1.0 eq), M04 (1.3 g, 6 mmol, 1.2 eq), Pd(OAc)2 (112 mg, 0.5 mmol, 0.1 eq), XantPhos (396 mg, 0.75 mmol, 0.15 eq), Cs2CO3 (2.44 g, 7.5 mmol, 1.5 eq), and toluene (50 mL) in sequence, heat the reaction system to 110 ° C, react for 24 h, concentrate by rotary evaporation, and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound B-4c-1 (white solid, 1.7 g), which is directly used in the next step.

[0221] Step 3: Weigh the compound B-4c-1 (159 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R71 (white solid, 100 mg). LCMS: [M+H] + =385. 1 H-NMR (500MHz, DMSO-d6) δ8.51(s,1H),7.46–7.41(m,3H),7.35(d,J=2.6Hz,1H),6.90(dd,J=8.4,2.6Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0222] Example 26: Synthesis of R73

[0223]

[0224] Step 1: In a 100 mL round-bottom flask, under nitrogen protection, compound 2d (3.0 g, 15 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.5 g, 18 mmol, 1.2 eq), Pd(PPh3)4 (3.5 g, 1.5 mmol, 0.1 eq), K3PO4 (8.0 g, 30 mmol, 2.0 eq), toluene / water (40 mL / 20 mL) were added successively, and the reaction system was heated to 80 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound M24 (white solid, 2.7 g).

[0225] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (1.5 g, 5 mmol, 1.0 eq), M24 (1.3 g, 6 mmol, 1.2 eq), Pd(OAc)2 (112 mg, 0.5 mmol, 0.1 eq), XantPhos (396 mg, 0.75 mmol, 0.15 eq), Cs2CO3 (2.44 g, 7.5 mmol, 1.5 eq), and toluene (50 mL) in sequence, heat the reaction system to 110 ° C, and react for 24 h. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound B-4d-1 (white solid, 1.9 g), which is directly used in the next step.

[0226] Step 3: Weigh the compound B-4d-1 (159 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R73 (white solid, 134 mg). LCMS: [M+H] + =377. 1 H-NMR (500MHz, DMSO-d6) δ8.51(s,1H),7.46–7.41(m,3H),7.35(d,J=2.6Hz,1H),6.90(dd,J=8.4,2.6Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0227] Example 27: Synthesis of R79

[0228]

[0229] Step 1: In a 100 mL round-bottom flask, under nitrogen protection, compound 2m (3.0 g, 15 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.5 g, 18 mmol, 1.2 eq), Pd(PPh3)4 (3.5 g, 1.5 mmol, 0.1 eq), K3PO4 (8.0 g, 30 mmol, 2.0 eq), toluene / water (40 mL / 20 mL) were added successively, and the reaction system was heated to 80 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound M05 (white solid, 2.7 g).

[0230] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (1.5 g, 5 mmol, 1.0 eq), M05 (1.3 g, 6 mmol, 1.2 eq), Pd(OAc)2 (112 mg, 0.5 mmol, 0.1 eq), XantPhos (396 mg, 0.75 mmol, 0.15 eq), Cs2CO3 (2.44 g, 7.5 mmol, 1.5 eq), and toluene (50 mL) in sequence, heat the reaction system to 110 ° C, react for 24 h, concentrate by rotary evaporation, and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound B-4m-1 (white solid, 1.9 g), which is directly used in the next step.

[0231] Step 3: Weigh the compound B-4m-1 (167 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R79 (white solid, 149 mg). LCMS: [M+H] + =405. 1 H-NMR (500MHz, DMSO-d6) δ8.51(s,1H),7.46–7.41(m,3H),7.35(d,J=2.6Hz,1H),6.90(dd,J=8.4,2.6Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0232] Example 28: Synthesis of R81

[0233]

[0234] Step 1: In a 100 mL round-bottom flask, under nitrogen protection, compound 2q (3.8 g, 15 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.5 g, 18 mmol, 1.2 eq), Pd(PPh3)4 (3.5 g, 1.5 mmol, 0.1 eq), K3PO4 (8.0 g, 30 mmol, 2.0 eq), toluene / water (40 mL / 20 mL) were added successively, and the reaction system was heated to 80 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound M25 (white solid, 2.7 g).

[0235] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (1.5 g, 5 mmol, 1.0 eq), M25 (1.6 g, 6 mmol, 1.2 eq), Pd(OAc)2 (112 mg, 0.5 mmol, 0.1 eq), XantPhos (396 mg, 0.75 mmol, 0.15 eq), Cs2CO3 (2.44 g, 7.5 mmol, 1.5 eq), and toluene (50 mL) in sequence, heat the reaction system to 110 ° C, and react for 24 h. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound B-4q-1 (white solid, 1.9 g), which is directly used in the next step.

[0236] Step 3: Weigh the compound B-4q-1 (177 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R81 (white solid, 158 mg). LCMS: [M+H] + =429. 1 H-NMR (500MHz, DMSO-d6) δ8.51(s,1H),7.46–7.41(m,3H),7.35(d,J=2.6Hz,1H),6.90(dd,J=8.4,2.6Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0237] Example 29: Synthesis of R86

[0238]

[0239] Step 1: In a 100 mL round-bottom flask, under nitrogen protection, compound 2f (3.1 g, 15 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.5 g, 18 mmol, 1.2 eq), Pd(PPh3)4 (3.5 g, 1.5 mmol, 0.1 eq), K3PO4 (8.0 g, 30 mmol, 2.0 eq), toluene / water (40 mL / 20 mL) were added successively, and the reaction system was heated to 80 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound M26 (white solid, 2.8 g).

[0240] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (1.5 g, 5 mmol, 1.0 eq), M26 (1.3 g, 6 mmol, 1.2 eq), Pd(OAc)2 (112 mg, 0.5 mmol, 0.1 eq), XantPhos (396 mg, 0.75 mmol, 0.15 eq), Cs2CO3 (2.44 g, 7.5 mmol, 1.5 eq), and toluene (50 mL) in sequence, heat the reaction system to 110 ° C, and react for 24 h. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound B-4f-1 (white solid, 1.6 g), which is directly used in the next step.

[0241] Step 3: Weigh the compound B-4f-1 (158 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), concentrated to dryness under reduced pressure, and the crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R86 (white solid, 158 mg).

[0242] LCMS: [M+H] + =381. 1 H-NMR (400MHz, DMSO-d6) δ9.87(d,J=3.5Hz,1H),9.15(d,J=3.8Hz,1H),7.80(dd,J=2.2,0.7Hz,1H),7.43(d,J=8.4Hz,1H),7.23–7.17(m,1H),7.11( d,J=2.2Hz,1H),7.08(dd,J=12.0,2.1Hz,1H),2.73–2.65(m,1H),2.41(d, J=10.3Hz,5H),1.65(dd,J=10.7,5.6Hz,2H),1.40(dd,J=10.7,5.6Hz,2H).

[0243] Example 30: Synthesis of R87

[0244]

[0245] Step 1: In a 25 mL round-bottom flask, compound R62 (83 mg, 0.2 mmol, 1.0 eq), HATU (114 mg, 0.3 mmol, 1.5 eq), N,N-dimethylformamide (2 mL) were weighed, stirred at room temperature for 5 min, and then diisopropylethylamine (77 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were added, heated to 45°C, and reacted for 16 h. The reaction solution was passed through a fast reverse phase column (water: acetonitrile = 1:13) to obtain the target compound R87 (white solid, 64 mg). LCMS: [M+H] + =416; 1H-NMR (500MHz, DMSO-d6) δ9.94(s,1H),8.11(s,1H),7.61(s,2H),7.45(s,1H),7.39(d,J=2.1Hz,1H),7.03(dd,J=8.7,2.1H z,1H),6.24(d,J=8.4Hz,1H),2.45(s,3H),2.27(s,3H),1.86–1.77(m,1H),0.92–0.81(m,2H),0.65(dt,J=6.5,3.2Hz,2H).

[0246] Example 31: Synthesis of R90

[0247]

[0248] Step 1: In a 100 mL round-bottom flask, under nitrogen protection, compound 2s (4.4 g, 15 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.5 g, 18 mmol, 1.2 eq), Pd(PPh3)4 (3.5 g, 1.5 mmol, 0.1 eq), K3PO4 (8.0 g, 30 mmol, 2.0 eq), toluene / water (40 mL / 20 mL) were added successively, and the reaction system was heated to 80 ° C and stirred for 24 h. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound M15 (white solid, 3.9 g).

[0249] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (1.5 g, 5 mmol, 1.0 eq), M15 (1.8 g, 6 mmol, 1.2 eq), Pd(OAc)2 (112 mg, 0.5 mmol, 0.1 eq), XantPhos (396 mg, 0.75 mmol, 0.15 eq), Cs2CO3 (2.44 g, 7.5 mmol, 1.5 eq), and toluene (50 mL) in sequence, heat the reaction system to 110 ° C, and react for 24 h. Concentrate by rotary evaporation and separate by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound B-4s-1 (white solid, 1.9 g), which is directly used in the next step.

[0250] Step 3: Weigh the compound B-4s-1 (192 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R90 (white solid, 174 mg). LCMS: [M+H] + =467. 1 H-NMR (500MHz, DMSO-d6) δ8.51(s,1H),7.46–7.41(m,3H),7.35(d,J=2.6Hz,1H),6.90(dd,J=8.4,2.6Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0251] Example 32: Synthesis of R218

[0252]

[0253] Step 1: In a 50 mL round-bottom flask, under nitrogen atmosphere, add B-3a (1.51 g, 5 mmol, 1.0 eq), 4-bromo-2-fluoro-6-methoxyaniline (1.3 g, 6 mmol, 1.2 eq), Pd(OAc)2 (112 mg, 0.5 mmol, 0.1 eq), XantPhos (396 mg, 0.75 mmol, 0.15 eq), Cs2CO3 (2.44 g, 7.5 mmol, 1.5 eq), and toluene (50 mL) in sequence, heat the reaction system to 110 ° C, and react for 24 h. Concentrate by rotary evaporation and separate by silica gel column chromatography (100% petroleum ether) to obtain compound B-4p (yellow solid, 2.4 g), which is directly used in the next step.

[0254] Step 2: In a 25 mL round-bottom flask, compound B-4p (197 mg, 0.5 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (85 mg, 0.6 mmol, 1.2 eq), Pd(dppf)Cl2 (37 mg, 0.05 mmol, 0.1 eq), K2CO3 (138 mg, 1.0 mmol, 2.0 eq), dioxane / water (2.5 mL / 0.25 mL, 10:1, v / v) were weighed and heated to 100 °C in a nitrogen atmosphere and stirred overnight. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound B-5p-1 (white solid, 164 mg), which was used directly in the next step.

[0255] Step 3: Weigh the compound B-5p-1 (164 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R218 (white solid, 143 mg). LCMS: [M+H] + =397; 1 H-NMR (500MHz, DMSO-d6) δ9.17(s,1H),7.60(d,J=2.3Hz,1H),7.07(dd,J=8.6,2.3Hz,1H),6.98(dd,J=11.0,1.8Hz,1H),6.93(d,J=1.7H z,1H),6.40(dd,J=8.6,4.2Hz,1H),3.84(s,3H),2.46(s,3H),2.29(s,3H),1.85(s,1H),0.93–0.81(m,2H),0.55(dd,J=5.0,1.9Hz,2H).

[0256] Example 33: Synthesis of R64

[0257]

[0258] Step 1: In a 25 mL round-bottom flask, compound B-4b (199 mg, 0.5 mmol, 1.0 eq), 3,5-dimethyl-1H-pyrazole-4-boronic acid (84 mg, 0.6 mmol, 1.2 eq), Pd(dppf)Cl2 (37 mg, 0.05 mmol, 0.1 eq), K2CO3 (138 mg, 1.0 mmol, 2.0 eq), dioxane / water (2.5 mL / 0.25 mL, 10:1, v / v) were weighed and heated to 100 °C in a nitrogen atmosphere and stirred overnight. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound B-5b-2 (white solid, 166 mg), which was used directly in the next step.

[0259] Step 2: Weigh the compound B-5b-2 (166 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R64 (white solid, 135 mg). LCMS: [M+H] + =400; 1 H-NMR (500MHz, DMSO-d6) δ8.51(s,1H),7.46–7.41(m,3H),7.35(d,J=2.6Hz,1H),6.90(dd,J=8.4,2.6Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0260] Example 34: Synthesis of R65

[0261]

[0262] In a 25 mL round-bottom flask, compound R64 (80 mg, 0.2 mmol, 1.0 eq), HATU (114 mg, 0.3 mmol, 1.5 eq), N,N-dimethylformamide (2 mL) were weighed, stirred at room temperature for 5 min, and then diisopropylethylamine (77 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were added, heated to 45°C, and reacted for 16 h. The reaction solution was passed through a fast reverse phase column (water: acetonitrile = 1:13) to obtain the target compound R65 (white solid, 64 mg). LCMS: [M+H] + =399. 1 H-NMR (500MHz, DMSO-d6) δ8.51(s,1H),7.46–7.41(m,3H),7.35(d,J=2.6Hz,1H),6.90(dd,J=8.4,2.6Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0263] Example 35: Synthesis of R66

[0264]

[0265] In a 50mL round-bottom flask, compound R64 (120mg, 0.3mmol, 1.0eq), NH2O-THP (18mg, 0.15mmol, 0.5eq), HOBT (49mg, 0.36mmol, 1.2eq), EDCI (69mg, 0.36mmol, 1.2eq), N,N-dimethylformamide (6mL) were weighed and stirred at room temperature overnight. The reaction solution was added with ice to precipitate solids, which were filtered, washed with water (2mL×3), washed with n-hexane (2mL×3), and dried to obtain the compound (white solid, 104mg), which was directly used in the next step. In a 50mL round-bottom flask, the product of the previous step (104mg, 0.21mmol, 1.0eq), trifluoroacetic acid (479mg, 4.2mmol, 20.0eq), dichloromethane / methanol (1.3mL / 1.3mL) were weighed and stirred at room temperature overnight. The reaction solution was directly filtered to obtain the target compound R66 (white solid, 55 mg). LCMS: [M+H] + =415. 1 H-NMR (500MHz, DMSO-d6) δ8.51(s,1H),7.46–7.41(m,3H),7.35(d,J=2.6Hz,1H),6.90(dd,J=8.4,2.6Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0266] Example 36: Synthesis of R74

[0267]

[0268] Step 1: In a 25 mL round-bottom flask, compound B-4b (199 mg, 0.5 mmol, 1.0 eq), 2-hydroxypyridine-4-boronic acid (83 mg, 0.6 mmol, 1.2 eq), Pd(dppf)Cl2 (37 mg, 0.05 mmol, 0.1 eq), K2CO3 (138 mg, 1.0 mmol, 2.0 eq), dioxane / water (2.5 mL / 0.25 mL, 10:1, v / v) were weighed and heated to 100 °C in a nitrogen atmosphere and stirred overnight. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound B-5b-8 (white solid, 166 mg), which was used directly in the next step.

[0269] Step 2: Weigh the compound B-5b-8 (172 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R74 (yellow solid, 138 mg). LCMS: [M+H] + =415. 1 H-NMR (500MHz, DMSO-d6) δ8.51(s,1H),7.46–7.41(m,3H),7.35(d,J=2.6Hz,1H),6.90(dd,J=8.4,2.6Hz,1H),3.80(s,3H),2.47(s,3H),2.40(s,3H).

[0270] Example 37: Synthesis of R127

[0271]

[0272] Step 1: In a 25 mL round-bottom flask, compound B-4b (199 mg, 0.5 mmol, 1.0 eq), pyridine-4-boric acid (74 mg, 0.6 mmol, 1.2 eq), Pd(dppf)Cl2 (37 mg, 0.05 mmol, 0.1 eq), K2CO3 (138 mg, 1.0 mmol, 2.0 eq), dioxane / water (2.5 mL / 0.25 mL, 10:1, v / v) were weighed and heated to 100 °C in a nitrogen atmosphere and stirred overnight. The mixture was concentrated by rotary evaporation and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound B-5b-4 (white solid, 166 mg), which was used directly in the next step.

[0273] Step 2: Weigh the compound B-5b-4 (165 mg, 0.4 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2.6 mL / 5.2 mL), add lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.3 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R127 (yellow solid, 145 mg). LCMS: [M+H] + =399; 1H-NMR(500MHz,DMSO-d6)δ9.61(s,1H),8.72–8.60(m,3H),8.08(s,3H),7.90–7.80(m,3H),7.64(d,J=2.3Hz ,1H),7.07(dd,J=8.6,2.3Hz,1H),6.24(d,J=8.5Hz,1H),1.86(s,1H),0.93–0.82(m,3H),0.63–0.47(m,3H).

[0274] Example 38: Synthesis of R77

[0275]

[0276] Step 1: In a 250 mL round-bottom flask, C-1a (5.0 g, 15 mmol, 1.0 eq), K2CO3 (2.1 g, 15 mmol, 1.0 eq), diethyl sulfate (2.3 g, 15 mmol, 1.0 eq), N,N-dimethylformamide (75 mL) were added in sequence and stirred at room temperature overnight. Water (75 mL) was added to the system, and the pH was adjusted to neutral with saturated ammonium chloride solution. The solution was washed with ethyl acetate (75 mL × 3), and the organic phases were combined. The organic phases were washed with water (75 mL × 3) and saturated brine (75 mL × 3) in sequence, and the organic phases were dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound C-2a (yellow oil, 5.0 g), which was directly used in the next step.

[0277] Step 2: In a 500 mL round-bottom flask, under nitrogen atmosphere, C-2a (5.0 g, 14 mmol, 1.2 eq), 4-bromo-2-fluoro-6-methylaniline (2.8 g, 11.7 mmol, 1.0 eq), Pd(OAc)2 (259 mg, 1.2 mmol, 0.1 eq), XantPhos (915 mg, 1.7 mmol, 0.15 eq), Cs2CO3 (5.6 g, 17 mmol, 1.5 eq), toluene (117 mL) were added in sequence, and the reaction system was heated to 110 ° C. and reacted for 24 h. The mixture was concentrated by rotary evaporation, and the crude product was separated by silica gel column chromatography (100% petroleum ether) to obtain compound C-3f (white solid, 4.8 g), which was directly used in the next step.

[0278] Step 3: In a 25 mL round-bottom flask, compound C-3f (436 mg, 1.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (211 mg, 1.5 mmol, 1.5 eq), Pd(dppf)Cl2 (73 mg, 0.1 mmol, 0.1 eq), K2CO3 (276 mg, 2.0 mmol, 2.0 eq), dioxane / water (5 mL / 0.5 mL) were added in sequence, heated to 100 ° C and stirred overnight in a nitrogen atmosphere. The reaction solution was concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound C-4f-1 (white solid, 234 mg), which was directly used in the next step.

[0279] Step 4: Weigh compound C-4f-1 (234 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3.2 mL / 6.4 mL), add lithium hydroxide (62 mg, 2.6 mmol, 5.0 eq) in water (1.6 mL), heat to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), concentrated to dryness under reduced pressure, and the crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R77 (white solid, 175 mg). LCMS: [M+H] + =425; 1 H-NMR(500MHz,DMSO-d6)δ13.71(s,1H),9.59(s,1H),7.80(d,J=2.9Hz,1H),7.70(s ,2H),7.42(dd,J=9.1,2.9Hz,1H),6.43(d,J=9.2Hz,1H),2.47(s,3H),2.29(s,3H).

[0280] Example 39: Synthesis of R161

[0281]

[0282] Step 1: In a 25 mL round-bottom flask, compound 2t (82 mg, 0.34 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (73 mg, 0.52 mmol, 1.5 eq), Pd(dppf)Cl2 (25.2 mg, 0.03 mmol, 0.1 eq), potassium carbonate (95.1 mg, 0.69 mmol, 2.0 eq), dioxane / water (4 mL / 0.4 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight in a nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound M32 (white solid, 45.5 mg).

[0283] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (64.7 mg, 0.2 mmol, 1.2 eq), M32 (45.5 mg, 0.18 mmol, 1.0 eq), Pd(OAc)2 (10 mg, 0.2 mmol, 0.1 eq), XantPhos (20 mg, 0.03 mmol, 0.15 eq), cesium carbonate (87.3 mg, 0.27 mmol, 1.5 eq), and toluene (5 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4t-1 (white solid, 107.5 mg).

[0284] Step 3: Add the compound B-4t-1 (53.7 mg, 0.1 mmol, 1.0 eq) of the previous step to tetrahydrofuran / ethanol (2 mL / 4 mL), add lithium hydroxide (12 mg, 0.5 mmol, 5.0 eq) in water (1 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R161 (white solid, 27.4 mg). LCMS: [M+H] + =415; 1 H-NMR (400MHz, DMSO-d6) δ13.21(s,1H),9.50(m,1H),7.64(d,J=2.2Hz,1H),7.31(t,J=10.2Hz,1H),7.13(dd,J=8.6,2.2Hz,1H ),6.57-6.21(m,1H),2.28(s,3H),2.15(s,3H),2.09(s,3H),1.89(qd,J=8.6,4.4Hz,1H),1.00-0.80(m,2H),0.65-0.35(m,2H).

[0285] Example 40: Synthesis of R162

[0286]

[0287] Step 1: In a 25 mL round-bottom flask, compound 2u (67 mg, 0.28 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (59.5 mg, 0.42 mmol, 1.5 eq), Pd(dppf)Cl2 (20.6 mg, 0.03 mmol, 0.1 eq), potassium carbonate (77.7 mg, 0.56 mmol, 2.0 eq), dioxane / water (4 mL / 0.4 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight in a nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound M49 (white solid, 30.5 mg).

[0288] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (30.5 mg, 0.10 mmol, 1.2 eq), M49 (45.5 mg, 0.08 mmol, 1.0 eq), Pd(OAc)2 (10 mg, 0.1 mmol, 0.1 eq), XantPhos (20 mg, 0.01 mmol, 0.15 eq), cesium carbonate (87.3 mg, 0.27 mmol, 1.5 eq), and toluene (5 mL) in sequence, heat the reaction system to 110 ° C, react for 24 h, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4u-1 (white solid, 35.1 mg).

[0289] Step 3: Add the compound B-4u-1 (35.1 mg, 0.08 mmol, 1.0 eq) in tetrahydrofuran / ethanol (2 mL / 4 mL), add lithium hydroxide (5.8 mg, 0.2 mmol, 5.0 eq) in water (0.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R162 (white solid, 11.9 mg). LCMS: [M+H] + =415; 1H-NMR (400MHz, DMSO-d6) δ13.21(s,1H),9.39(s,1H),7.65(d,J=2.2Hz,1H),7.36(t,J=4.1Hz,1H),7.12(dd,J=8.6,2.3Hz,1H),6.4 6(dd,J=8.6,4.1Hz,1H),2.28(s,3H),2.10(s,3H),2.03(d,J=2.7Hz,3H),1.93-1.84(m,1H),0.92-0.84(m,2H),0.61-0.55(m,2H).

[0290] Example 41: Synthesis of R154

[0291]

[0292] Step 1: In a 100 mL round-bottom flask, compound 2b (2.00 g, 8.89 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (1.88 g, 13.3 mmol, 1.5 eq), Pd(dppf)Cl2 (651 mg, 0.9 mmol, 0.1 eq), potassium carbonate (2.45 g, 17.8 mmol, 2.0 eq), dioxane / water (30 mL / 3 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight in a nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound M33 (white solid, 1.94 g).

[0293] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, M33 (100 mg, 0.41 mmol, 1.0 eq), 2-methoxy-5-bromopyridine-4-carboxylic acid methyl ester (122 mg, 0.49 mmol, 1.2 eq), Pd(OAc)2 (10 mg, 0.04 mmol, 0.1 eq), XantPhos (36 mg, 0.06 mmol, 0.15 eq), cesium carbonate (203 mg, 0.62 mmol, 1.5 eq), and toluene (4 mL) were added in sequence, and the reaction system was heated to 110 ° C for 24 h, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound D-4b-1 (yellow solid, 144 mg).

[0294] Step 3: Add the compound D-4b-1 (90 mg, 0.22 mmol, 1.0 eq) of the previous step to tetrahydrofuran / ethanol (2 mL / 4 mL), add lithium hydroxide (16 mg, 0.66 mmol, 5.0 eq) in water (1 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was washed with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R154 (yellow solid, 68.4 mg). LCMS: [M+H] + =392; 1 H-NMR (400MHz, DMSO-d6) δ8.56 (s, 1H), 7.60 (d, J = 3.9Hz, 1H), 7.50 (s, 1H), 7.43 (d ,J=11.5Hz,1H),7.19(s,1H),5.76(s,1H),3.81(s,3H),2.46(s,3H),2.29(s,3H).

[0295] Example 42: Synthesis of R157

[0296]

[0297] Step 1: In a 100 mL round-bottom flask, add E01 (600 mg, 3.8 mmol, 1.0 eq), deuterated iodomethane (1.00 g, 6.8 mmol, 3.0 eq), potassium carbonate (2.09 g, 9.1 mmol, 4.0 eq), and DMF (10 mL) in sequence, heat to 40 ° C and stir overnight. After the reaction is completed, filter. The filtrate is concentrated to dryness to obtain E02 (white oil, 834 mg), which is directly used in the next step.

[0298] Step 2: In a 50 mL round-bottom flask, under nitrogen atmosphere, E02 (186.7 mg, 0.62 mmol, 1.0 eq), M33 (100 mg, 0.41 mmol, 1.2 eq), Pd(OAc)2 (10 mg, 0.04 mmol, 0.1 eq), XantPhos (36 mg, 0.06 mmol, 0.15 eq), cesium carbonate (203 mg, 0.62 mmol, 1.5 eq), toluene (4 mL) were added in sequence, and the reaction system was heated to 110°C and reacted for 24 h. After concentration, the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound E-03-1 (white solid, 88.8 mg).

[0299] Step 3: Add the compound E-03-1 (49.5 mg, 0.12 mmol, 1.0 eq) of the previous step to tetrahydrofuran / ethanol (2 mL / 4 mL), add lithium hydroxide (8.7 mg, 0.36 mmol, 3.0 eq) in water (1 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was washed with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R157 (white solid, 34.5 mg). LCMS: [M+H] + =394; 1 H-NMR(400MHz,DMSO-d6)δ9.26(s,1H),7.50(s,1H),7.48-7.39(m,2H),7.07( dd,J=9.1,3.1Hz,1H),6.53(dd,J=9.0,4.4Hz,1H),2.46(s,3H),2.29(s,3H).

[0300] Example 43: Synthesis of R164

[0301]

[0302] Step 1: In a 100 mL round-bottom flask, compound 2ac (518.0 mg, 2.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (422.2 mg, 3.0 mmol, 1.5 eq), Pd(dppf)Cl2 (146.3 mg, 0.2 mmol, 0.1 eq), potassium carbonate (276.4 mg, 2.0 mmol, 2.0 eq), dioxane / water (30 mL / 3 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight under nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound M50 (white solid, 350 mg).

[0303] Step 2: Under nitrogen protection, add M50 (324.4 mg, 1.18 mmol, 1.0 eq) and THF (20 mL) to a 50 mL three-necked flask equipped with a mechanical stirrer, a thermometer, and a constant pressure dropping funnel, cool to -78 ° C, add n-butyl lithium (1 ml, 1.53 mmol, 1.3 eq) dropwise, and react at -78 ° C for 1 h. Dissolve iodine (600 mg, 2.36 mmol, 2.0 eq) in THF (2 mL) and add dropwise to the above reaction system. After 2 h of warm reaction, quench the reaction with water, wash the solution with ethyl acetate (100 mL), combine the organic phases, wash with saturated sodium chloride solution (20 mL), dry with anhydrous sodium sulfate, and filter. Concentrate to dryness under reduced pressure. The crude product is separated by column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound M50-1 (white solid, 293.6 mg).

[0304] Step 3: In a 25 mL round-bottom flask, under a nitrogen atmosphere, M50-1 (90.8 mg, 0.23 mmol, 1.2 eq), B-2a (36 mg, 0.19 mmol, 1.0 eq), Pd(OAc)2 (10 mg, 0.02 mmol, 0.1 eq), XantPhos (16.4 mg, 0.03 mmol, 0.15 eq), cesium carbonate (92.3 mg, 0.28 mmol, 1.5 eq), and toluene (4 mL) were added in sequence, the reaction system was heated to 110 ° C., reacted for 24 hours, and concentrated to dryness. The crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 50: 1) to obtain compound B-4ad-1 (white solid, 54.5 mg).

[0305] Step 4: Add the compound B-4ad-1 (54.5 mg, 0.12 mmol, 1.0 eq) of the previous step to tetrahydrofuran / ethanol (2 mL / 4 mL), add lithium hydroxide (14.4 mg, 0.60 mmol, 5.0 eq) in water (1 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was washed with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R164 (white solid, 6.9 mg). LCMS: [M+H] + =451; 1H-NMR (400MHz, DMSO-d6) δ9.54(s,1H),7.64(d,J=2.2Hz,1H),7.54(dd,J=11.2,1.8Hz,1H),7.43(s,1H),7.14(dd,J=8.6,2.3Hz,1H ),6.52(dd,J=8.6,4.6Hz,1H),2.47(s,3H),2.30(s,3H),1.89(ddd,J=13.5,8.5,5.1Hz,1H),0.94-0.81(m,2H),0.68-0.52(m,2H).

[0306] Example 44: Synthesis of R165

[0307]

[0308] Step 1: In a 500mL round-bottom flask, add M33 (1.00g, 4.2mmol, 1.0eq), water (10mL), concentrated hydrochloric acid (12M, 10mL), and slowly add sodium nitrite (345mg, 5.0mmol, 1.2eq) aqueous solution (2mL) in an ice-water bath. After the addition, continue to keep stirring at 0℃ for 0.5h. Continue to slowly add potassium iodide (1.0g, 6.3mmol, 1.5eq) aqueous solution (5mL) in a constant pressure dropping funnel. After the addition, heat to 40℃ and stir for 10min. After the reaction is completed, add saturated sodium bicarbonate aqueous solution to quench, wash the solution with ethyl acetate (200mL), combine the organic phases, wash with saturated sodium chloride solution (100mL), dry over anhydrous sodium sulfate, and filter. The filtrate is concentrated, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain M33-1 (white solid, 766mg).

[0309] Step 2: In a 25 mL round-bottom flask, under a nitrogen atmosphere, M33-1 (13.7 mg, 0.08 mmol, 1.0 eq), methyl 2-amino-5-ethylbenzoate (32.3 mg, 0.09 mmol, 1.2 eq), Pd(OAc)2 (10 mg, 0.01 mmol, 0.1 eq), XantPhos (10 mg, 0.01 mmol, 0.15 eq), cesium carbonate (37.4 mg, 0.11 mmol, 1.5 eq), and toluene (4 mL) were added in sequence, and the reaction system was heated to 110 ° C., reacted for 24 hours and concentrated. The crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain compound F-4b-1 (white solid, 10.8 mg).

[0310] Step 3: Add the compound F-4b-1 (10.8 mg, 0.03 mmol, 1.0 eq) of the previous step to tetrahydrofuran / ethanol (1 mL / 2 mL), add lithium hydroxide (7.2 mg, 0.3 mmol, 10.0 eq) in water (1 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was washed with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R165 (white solid, 7.7 mg). LCMS: [M+H] + =389; 1 H-NMR (400MHz, DMSO-d6) δ13.20(s,1H),9.54(s,1H),7.75(d,J=2.1Hz,1H),7.55-7.50(m,1H),7.44(dt,J=24.1,12.0Hz,1H), 7.29-7.21(m,1H),6.47(dd,J=8.5,4.3Hz,1H),2.61-2.53(m,3H),2.47(s,3H),2.31(d,J=18.5Hz,2H),1.16(t,J=7.6Hz,3H).

[0311] Example 45: Synthesis of R168

[0312]

[0313] Step 1: In a 25 mL round-bottom flask, add compound 2u (54.9 mg, 0.23 mmol, 1.0 eq), 3,5-dimethylpyrazole-4-boronic acid pinacol ester (77 mg, 0.35 mmol, 1.5 eq), Pd(dppf)Cl2 (16.9 mg, 0.02 mmol, 0.1 eq), potassium carbonate (63.7 mg, 0.46 mmol, 2.0 eq), dioxane / water (4 mL / 0.4 mL, 10:1, v / v), heat to 95 ° C and stir overnight in a nitrogen atmosphere. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound M47 (white solid, 52.9 mg), which is directly used in the next step.

[0314] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, B-3a (58.3 mg, 0.19 mmol, 1.2 eq), M47 (52.9 mg, 0.16 mmol, 1.0 eq), Pd(OAc)2 (10 mg, 0.02 mmol, 0.1 eq), XantPhos (15 mg, 0.03 mmol, 0.15 eq), cesium carbonate (78.7 mg, 0.24 mmol, 1.5 eq), and toluene (4 mL) were added in sequence, and the reaction system was heated to 110 ° C., reacted for 24 hours and concentrated. The crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4u-2 (yellow oil, 36.4 mg), which was directly used in the next step.

[0315] Step 3: Add the compound B-4u-2 (32.4 mg, 0.07 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (2 mL / 4 mL), add lithium hydroxide (10.1 mg, 0.42 mmol, 6.0 eq) in water (1 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was washed with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R168 (white solid, 5.4 mg). LCMS: [M+H] + =414; 1 H-NMR (400MHz, DMSO-d6) δ12.76(s,2H),9.36(s,1H),7.64(d,J=2.2Hz,1H),7.18-7.16(m,1H),7.14-7.08(m,1H),6.41(dd,J= 8.6,3.7Hz,1H),2.04(s,6H),2.01(d,J=2.8Hz,3H),1.88(ddd,J=13.5,8.6,5.2Hz,1H),0.90-0.85(m,2H),0.60-0.53(m,2H).

[0316] Example 46: Synthesis of R79

[0317]

[0318] Step 1: In a 250 mL round-bottom flask, compound 2m (2.28 g, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight in a nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M05 (white solid, 1.50 g).

[0319] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M05 (196 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4m-1 (white solid, 256 mg).

[0320] Step 3: Add the compound B-4m-1 (209 mg, 0.5 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (3 mL / 6 mL), add a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R79 (white solid, 154 mg). LCMS: [M+H] + =405; 1HNMR(400MHz,DMSO-d6)δ9.13(s,1H),7.61(d,J=2.3Hz,1H),7.20(s,2H),7.02(dd,J=8.3,2.1Hz,1H),6.08(d,J=8.5Hz,1H),2.45(s,3H),2 .28(s,3H),2.01(q,J=7.2Hz,1H),1.82(tt,J=8.9,5.1Hz,1H),1.24(s,2H),1.09(t,J=7.5Hz,6H),0.88–0.81(m,2H),0.53(q,J=5.3Hz,2H).

[0321] Example 47: Synthesis of R207

[0322]

[0323] Step 1: In a 250 mL round-bottom flask, compound 2b (2.24 g, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v), heated to 95 ° C and stirred overnight in a nitrogen atmosphere. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M33 (white solid, 1.25 g).

[0324] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M33 (193 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4b-1 (white solid, 326 mg).

[0325] Step 3: Add the compound B-4b-1 (208 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R207 (white solid, 192 mg). LCMS: [M+H] + =401; 1 H NMR (400MHz, DMSO-d6) δ9.43 (s, 1H), 7.64 (d, J = 2.3Hz, 1H), 7.51 (s, 1H), 7.45 (d, J = 11.2Hz, 1H), 7.13 (dd, J = 8.4, 2.2Hz, 1H), 6.45 (dd, J = 8.7, 4.1Hz, 1H),2.47(s,3H),2.29(s,3H),2.08(s,1H),2.03–1.94(m,1H),1.88(td,J= 8.6, 4.4Hz, 1H), 1.24 (s, 4H), 0.88 (p, J = 6.6Hz, 2H), 0.59 (t, J = 5.2Hz, 2H).

[0326] Example 48: Synthesis of R208

[0327]

[0328] Step 1: In a 250 mL round-bottom flask, add compound 2h (2562 mg, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v), and heat to 95 ° C in a nitrogen atmosphere and stir overnight. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M28 (white solid, 1.46 g).

[0329] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M28 (218 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 h, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4h-1 (white solid, 386 mg).

[0330] Step 3: Add the compound B-4h-1 (208 mg, 0.4 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (3 mL / 6 mL), add a solution of lithium hydroxide (48 mg, 2.0 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R208 (white solid, 192 mg). LCMS: [M+H] + =433; 1 H NMR (400MHz, DMSO-d6) δ9.09(s,1H),7.60(t,J=1.8Hz,1H),7.23(s,2H),7.07–6.99(m,1H),6.07(d,J=8.6Hz,1H),3.04(p,J=6.9Hz,2H),2.47(s,3H ),2.30(s,3H),2.06–1.93(m,1H),1.82(tt,J=8.8,5.1Hz,1H),1.24(s,2H ),1.13(dd,J=15.4,6.8Hz,11H),0.89–0.79(m,2H),0.54(t,J=5.3Hz,2H).

[0331] Example 49: Synthesis of R90

[0332]

[0333] Step 1: In a 250 mL round-bottom flask, compound 2s (2905 mg, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight under nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M46 (white solid, 2106 mg).

[0334] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M46 (246 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4s-1 (white solid, 356 mg).

[0335] Step 3: Add the compound B-4s-1 (208 mg, 0.7 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (4 mL / 8 mL), add a solution of lithium hydroxide (84 mg, 3.5 mmol, 5.0 eq) in water (2.0 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R90 (white solid, 115 mg). LCMS: [M+H] + =467; 1HNMR(400MHz,DMSO-d6)δ9.55(s,1H),7.74(s,1H),7.64(s,1H),7.56(s,1H),7.11(d,J=8.5Hz,1H),6.37(d,J=8.7Hz,1H),2.4 7(d,J=1.7Hz,3H),2.29(d,J=1.8Hz,3H),2.00(s,1H),1.88(s,1H),1.24(s,3H),0.88(t,J=7.4Hz,2H),0.58(d,J=5.4Hz,2H).

[0336] Example 50: Synthesis of R210

[0337]

[0338] Step 1: In a 250 mL round-bottom flask, compound 2e (2205 mg, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight under nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M36 (white solid, 1.65 g).

[0339] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M36 (190 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound B-4e-1 (white solid, 314 mg).

[0340] Step 3: Add the compound B-4e-1 (288 mg, 0.7 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (4 mL / 8 mL), add a solution of lithium hydroxide (84 mg, 3.5 mmol, 5.0 eq) in water (2.0 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R210 (white solid, 192 mg). LCMS: [M+H] + =397; 1 H NMR (600MHz, DMSO-d6) δ13.04(s,1H),9.23(s,1H),7.64(d,J=2.4Hz,1H),7.47(s,1H),7.35(s,1H),7.06(d,J=8.6Hz,1H),6.15 (d,J=8.3Hz,1H),2.45(s,3H),2.27(s,3H),2.22(s,3H),1.85(tt,J=8.8,4.9Hz,1H),0.89–0.84(m,2H),0.55(d,J=5.3Hz,2H).

[0341] Example 51: Synthesis of R211

[0342]

[0343] Step 1: In a 250 mL round-bottom flask, compound 2v (2.34 g, 10.0 mmol, 1.0 eq), 3,5-3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight in a nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M37 (white solid, 1.46 mg).

[0344] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M37 (201 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 h, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound B-4v-1 (white solid, 300 mg).

[0345] Step 3: Add the compound B-4v-1 (221 mg, 0.5 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (3 mL / 6 mL), add a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R211 (white solid, 144 mg). LCMS: [M+H] + =411; 1 H NMR(600MHz,DMSO-d6)δ13.05(s,1H),9.24(s,1H),7.63(d,J=2.3Hz,1H),7.48 (d,J=2.3Hz,1H),7.36(d,J=2.5Hz,1H),7.06(dd,J=8.5,2.6Hz,1H),6.17–6.10 (m,1H),2.58(q,J=7.7Hz,2H),2.46(d,J=1.7Hz,3H),2.28(d,J=1.7Hz,3H),1.8 8–1.81(m,1H),1.14–1.08(m,3H),0.86(d,J=8.0Hz,2H),0.55(d,J=5.2Hz,2H).

[0346] Example 52: Synthesis of R212

[0347]

[0348] Step 1: In a 250 mL round-bottom flask, add compound 2g (2.75 mg, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v), and heat to 95°C in a nitrogen atmosphere and stir overnight. Concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M11 (white solid, 1.37 g).

[0349] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M11 (233 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4g-1 (white solid, 315 mg).

[0350] Step 3: Add the compound B-4g-1 (279 mg, 0.6 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (3 mL / 6 mL), add a solution of lithium hydroxide (72 mg, 3.0 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R212 (white solid, 135 mg). LCMS: [M+H] + =451; 1H NMR (600MHz, DMSO-d6) δ13.19(s,1H),9.57(s,1H),7.98(s,1H),7.79(s,1H),7.64(d,J=2.5Hz,1H),7.08(d,J=8.7Hz,1H),6.22 (d,J=8.6Hz,1H),2.47(d,J=1.8Hz,3H),2.29(d,J=1.7Hz,3H),1.90–1.84(m,1H),0.88(d,J=8.1Hz,2H),0.57(d,J=5.2Hz,2H).

[0351] Example 53: Synthesis of R213

[0352]

[0353] Step 1: In a 250 mL round-bottom flask, compound 2j (2.36 g, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, heated to 95 °C and stirred overnight under nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M38 (white solid, 1.69 g).

[0354] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M38 (203 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), toluene (8 mL, heat the reaction system to 110 ° C, react for 24 h, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4j-1 (white solid, 331 mg).

[0355] Step 3: Add the compound B-4j-1 (221 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R213 (white solid, 138 mg). LCMS: [M+H] + =413; 1 H NMR (600MHz, DMSO-d6) δ13.01(s,1H),9.15(s,1H),7.60(d,J=2.8Hz,1H),7.20(d,J=2.0Hz,1H),7.11–7.03(m,2H),6.26(d, J=8.8Hz,1H),3.81(s,3H),2.47(s,3H),2.29(s,3H),1.86(dd,J=8.9,4.4Hz,1H),0.90–0.83(m,2H),0.55(d,J=5.2Hz,2H).

[0356] Example 54: Synthesis of R214

[0357]

[0358] Step 1: In a 250 mL round-bottom flask, compound 2w (2.50 g, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight under nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M39 (white solid, 2.02 g).

[0359] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M39 (214 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 h, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4w-1 (white solid, 245 mg).

[0360] Step 3: Add the compound B-4w-1 (221 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R214 (white solid, 144 mg). LCMS: [M+H] + =427; 1 H NMR (600MHz, DMSO-d6) δ13.01(s,1H),9.22(s,1H),7.60(d,J=2.7Hz,1H),7.17(s,1H),7.10–7.02(m,2H),6.31(d,J=8.7Hz,1H),4.10(q,J=7.0Hz,2 H),3.31(s,2H),2.46(s,3H),2.28(d,J=2.0Hz,3H),1.87(d,J=6.7Hz,1H) ,1.18(t,J=6.9Hz,3H),0.87(dq,J=8.8,3.2Hz,2H),0.56(d,J=5.2Hz,2H).

[0361] Example 55: Synthesis of R215

[0362]

[0363] Step 1: In a 250 mL round-bottom flask, compound 2o (2.28 g, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight in a nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M40 (white solid, 1.71 g).

[0364] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M40 (196 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4o-1 (white solid, 227 mg).

[0365] Step 3: Add the compound B-4o-1 (210 mg, 0.5 mmol, 1.0 eq) in tetrahydrofuran / ethanol (3 mL / 6 mL), add lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R215 (white solid, 125 mg). LCMS: [M+H] + =405; 1H NMR (600MHz, DMSO-d6) δ12.88(s,1H),9.10(s,1H),7.62(d,J=2.7Hz,1H),7.21(d,J=11. 2Hz,2H),7.03(d,J=8.7Hz,1H),6.07(d,J=8.4Hz,1H),3.31(s,2H),3.11–3.05(m,1H),2 .45(d,J=2.0Hz,3H),2.28(d,J=1.9Hz,3H),2.13(s,3H),1.82(d,J=7.7Hz,1H),1.16(d, J=6.9Hz,3H),1.12(d,J=6.8Hz,3H),0.85(dq,J=8.8,3.1Hz,2H),0.54(d,J=5.1Hz,2H).

[0366] Example 56: Synthesis of R216

[0367]

[0368] Step 1: In a 250 mL round-bottom flask, compound 2x (2.46 g, 10.0 mmol, 1.0 eq), dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, and heated to 95 ° C. and stirred overnight in a nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M41 (white solid, 1.47 g).

[0369] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M41 (211 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 h, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound B-4x-1 (white solid, 296 mg).

[0370] Step 3: Add the compound B-4x-1 (263 mg, 0.6 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (3 mL / 6 mL), add a solution of lithium hydroxide (72 mg, 3.0 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R216 (white solid, 125 mg). LCMS: [M+H] + =423; 1 H NMR (600MHz, DMSO-d6) δ13.04(s,1H),9.36(s,1H),7.63(d,J=2.8Hz,1H),7.42(d,J=2.6Hz,1H),7.08(d,J=8.3Hz,1H),6.89(s,1H),6.27(d,J= 8.5Hz,1H),2.43(s,3H),2.25(s,3H),1.91(t,J=6.9Hz,1H),1.85(q,J= 6.8Hz,1H),0.94–0.85(m,4H),0.79–0.67(m,2H),0.56(d,J=5.4Hz,2H).

[0371] Example 57: Synthesis of R217

[0372]

[0373] Step 1: In a 250 mL round-bottom flask, compound 2y (2.15 g, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, heated to 95 °C and stirred overnight under nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M42 (white solid, 1.16 g).

[0374] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M42 (185 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4y-1 (white solid, 297 mg).

[0375] Step 3: Add the compound B-4y-1 (284 mg, 0.7 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (4 mL / 8 mL), add a solution of lithium hydroxide (84 mg, 3.5 mmol, 5.0 eq) in water (2.0 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R217 (white solid, 185 mg). LCMS: [M+H] + =392; 1 H NMR (600MHz, DMSO-d6) δ13.32(s,1H),9.64(s,1H),7.82–7.75(m,2H),7.66(d,J=2.3Hz,1H),7.17(dd,J=8.6,2.3Hz, 1H),6.64(dd,J=8.6,3.8Hz,1H),2.47(s,3H),2.29(s,3H),1.96–1.87(m,1H),0.95–0.86(m,2H),0.66–0.54(m,2H).

[0376] Example 58: Synthesis of R219

[0377]

[0378] Step 1: In a 250 mL round-bottom flask, compound 2z (2.48 g, 10.0 mmol, 1.0 eq), dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight in a nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M43 (white solid, 1.84 g).

[0379] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M43 (212 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound B-4z-1 (white solid, 205 mg).

[0380] Step 3: Add the compound B-4z-1 (176 mg, 0.4 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (3 mL / 6 mL), add a solution of lithium hydroxide (84 mg, 2.0 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R219 (white solid, 102 mg). LCMS: [M+H] + =425; 1H NMR (600MHz, DMSO-d6) δ13.02(s,1H),9.17(s,1H),7.61(d,J=2.3Hz,1H),7.12(dd,J=8.7,2.3Hz,1H),7.02–6.90(m,2H),6.48(dd,J=8.6,4.8Hz, 1H),4.67(hept,J=6.1Hz,1H),2.46(s,3H),2.29(s,3H),1.87(tt,J=8.4 ,5.1Hz,1H),1.23(d,J=6.0Hz,6H),0.90–0.85(m,2H),0.60–0.53(m,2H).

[0381] Example 59: Synthesis of R220

[0382]

[0383] Step 1: In a 250 mL round-bottom flask, compound 2aa (2.18 g, 10.0 mmol, 1.0 eq), dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL, 10:1, v / v) were added, heated to 95 ° C and stirred overnight under nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M44 (white solid, 1.09 g).

[0384] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol, 1.2 eq), M44 (188 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9: 1) to obtain compound B-4aa-1 (white solid, 220 mg).

[0385] Step 3: Add the compound B-4aa-1 (205 mg, 0.5 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (3 mL / 6 mL), add a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was extracted with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R220 (white solid, 145 mg). LCMS: [M+H] + =395;

[0386] 1 H NMR (600MHz, DMSO-d6) δ13.06 (s, 1H), 9.17 (s, 1H), 7.63 (d, J = 2.3Hz, 1H), 7.25 ( dd,J=11.1,2.0Hz,1H),7.20(d,J=2.0Hz,1H),7.09(dd,J=8.6,2.3Hz,1H),6.32( dd,J=8.6,3.2Hz,1H),2.62(q,J=7.6Hz,2H),2.46(s,3H),2.29(s,3H),1.85(td ,J=8.5,4.3Hz,1H),1.13(t,J=7.5Hz,3H),0.92–0.81(m,2H),0.61–0.51(m,2H).

[0387] Example 60: Synthesis of R221

[0388]

[0389] Step 1: In a 250 mL round-bottom flask, compound 2bb (5000 mg, 20.7 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (4388 mg, 31.1 mmol, 1.5 eq), Pd(dppf)Cl2 (1519 mg, 2.1 mmol, 0.1 eq), potassium carbonate (5726 mg, 41.5 mmol, 2.0 eq), dioxane / water (30 mL / 3 mL, 10:1, v / v) were added, heated to 95 °C and stirred overnight under nitrogen atmosphere. The crude product was concentrated and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound M45 (white solid, 5 g).

[0390] Step 2: Under nitrogen protection, add M45 (5g, 19.5mmol, 1.0eq) and THF (100mL) to a 250mL three-necked flask equipped with a mechanical stirrer, a thermometer, and a constant pressure dropping funnel, cool to -78°C, add n-butyl lithium (18ml, 29.2mmol, 1.5eq) dropwise, and react for 1h after the addition is complete. Dissolve iodine (7907mg, 31.1mmol, 1.6eq) in THF (10mL) and add dropwise to the above reaction system, and react for 2 hours. Quench the reaction with water, wash the solution with ethyl acetate (100mL), combine the organic phases, wash the organic phases with saturated sodium chloride solution (20mL), dry with anhydrous sodium sulfate, and filter. Concentrate to dryness under reduced pressure. The crude product is separated by column chromatography (petroleum ether: ethyl acetate = 40:1) to obtain compound M45-1 (white solid, 3.2g).

[0391] Step 3: In a 100 mL round-bottom flask, under a nitrogen atmosphere, add M45-1 (3.2 g, 8.4 mmol, 1.0 eq), B-2a (1929 mg, 10.1 mmol, 1.2 eq), Pd(OAc)2 (189 mg, 0.8 mmol, 0.1 eq), XantPhos (727.6 mg, 1.3 mmol, 0.15 eq), cesium carbonate (4097 mg, 12.6 mmol, 1.5 eq), and toluene (40 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, and concentrate. The crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 50: 1) to obtain compound B-4ac-1 (yellow oil, 1.6 g).

[0392] Step 4: Add the compound B-4ac-1 (1.6 g, 3.6 mmol, 1.0 eq) from the previous step to tetrahydrofuran / ethanol (10 mL / 20 mL), add a solution of lithium hydroxide (525 mg, 21.9 mmol, 6.0 eq) in water (5 mL), heat to 45 ° C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and the solution was washed with (dichloromethane: methanol = 10: 1) (50 mL), the organic phases were combined, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R221 (white solid, 1.4 g). LCMS: [M+H] + =433;

[0393] 1H-NMR (400MHz, DMSO-d6) δ13.17(s,1H),9.26(s,1H),7.63(d,J=2.3Hz,1H),7.37(dd,J=11.0,1.7Hz,1H),7.20(s,1H),7.12(dd,J =8.7,2.3Hz,1H),6.48(dd,J=8.6,4.3Hz,1H),2.47(s,3H),2.29(s,3H),2.03–1.68(m,1H),0.95–0.76(m,2H),0.65–0.44(m,2H).

[0394] Example 61: Synthesis of R240

[0395]

[0396] Step 1: In a 250 mL round-bottom flask, add compound R240-1 (2.18 g, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL), heat to 95 ° C in a nitrogen atmosphere and stir overnight. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound R240-2 (white solid, 1.09 g).

[0397] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, methyl 2-iodo-5-methylbenzoate (302 mg, 1.0 mmol), R240-2 (188 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) were added in sequence, and the reaction system was heated to 110 ° C., reacted for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R240-3 (white solid, 220 mg).

[0398] Step 3: Compound R240-3 (205 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45 °C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R240-4 (white solid, 145 mg).

[0399] Step 4: Compound R240-4 (78 mg, 0.2 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (column model: Spherical C18, 20-45 μM; mobile phase water: acetonitrile = 1:5) to obtain compound R240 (white solid, 56 mg). LCMS: [M+H] + =386; 1 H NMR(500MHz,DMSO-d6)δ8.23(s,1H),7.66–7.62(m,1H),7.30–7.23(m,1H),7.19–7.1 3(m,1H),7.07(d,J=2.2Hz,1H),6.74(s,1H),3.89(s,2H),2.55(s,2H),2.42(s,2H).

[0400] Example 62: Synthesis of R241

[0401]

[0402] Step 1: In a 50 mL round-bottom flask, under a nitrogen atmosphere, B-3a (303 mg, 1.0 mmol, 1.2 eq), R240-2 (205 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) were added in sequence, and the reaction system was heated to 110 ° C for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R241-1 (white solid, 245 mg).

[0403] Step 2: Compound R241-1 (214 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R241-2 (white solid, 178 mg).

[0404] Step 3: Compound R241-2 (83 mg, 0.2 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (water: acetonitrile = 1:5) to obtain compound R241 (white solid, 56 mg). LCMS: [M+H] + =412; 1 H NMR(500MHz,DMSO-d6)δ8.34(s,1H),7.81(dd,J=2.1,0.7Hz,1H),7.37(d,J=8.5Hz,1H),7.29(s,0H),7.20–7.1 4(m,1H),7.09–7.05(m,3H),3.89(s,2H),2.73–2.65(m,1H),2.55(s,2H),1.73–1.65(m,2H),1.48–1.40(m,2H).

[0405] Example 63: Synthesis of R242

[0406]

[0407] Step 1: In a 250 mL round-bottom flask, add compound R242-1 (2.14 g, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL), heat to 95 ° C and stir overnight in a nitrogen atmosphere. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound R242-2 (white solid, 0.98 g).

[0408] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (303 mg, 1.0 mmol, 1.2 eq), R242-2 (185 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R242-3 (white solid, 231 mg).

[0409] Step 3: Compound R242-3 (202 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45 °C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R242-4 (white solid, 155 mg).

[0410] Step 4: Compound R242-4 (78 mg, 0.2 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (column model: Spherical C18, 20-45 μM; mobile phase water: acetonitrile = 1:5) to obtain compound R242 (white solid, 44 mg). LCMS: [M+H] + =390; 1 H NMR (500MHz, DMSO-d6) δ7.81(dd,J=2.1,0.8Hz,1H),7.68(s,1H),7.33(s,1H),7.21(dt,J=1.8,0.9Hz,1H),7.20–7.14(m,1H),7.10(d,J=2.6Hz,1H) ,7.07(s,1H),2.73–2.65(m,1H),2.59(qd,J=7.1,0.9Hz,2H),2.55(s,2H) ,2.42(s,2H),1.73–1.65(m,2H),1.48–1.40(m,2H),1.25(t,J=7.1Hz,3H).

[0411] Example 64: Synthesis of R243

[0412]

[0413] Step 1: In a 50 mL round-bottom flask, under a nitrogen atmosphere, methyl 2-iodo-5-methylbenzoate (302 mg, 1.0 mmol, 1.2 eq), R242-2 (185 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol), and toluene (8 mL) were added in sequence, and the reaction system was heated to 110 ° C., reacted for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R243-1 (white solid, 216 mg).

[0414] Step 2: Compound R240-1 (190 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R243-2 (white solid, 143 mg).

[0415] Step 3: Compound R243-2 (73 mg, 0.2 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (column model: Spherical C18, 20-45 μM; mobile phase water: acetonitrile = 1:5) to obtain compound R243 (white solid, 51 mg). LCMS: [M+H] + =364; 1 H NMR (500MHz, DMSO-d6) δ7.66–7.62(m,2H),7.58(s,1H),7.28–7.14(m,4H),7.10(d,J=2.6Hz,1H),6.76(s,2 H),2.59(qd,J=7.1,0.9Hz,3H),2.55(s,4H),2.42(s,4H),2.37(s,4H),2.26(s,5H),1.25(t,J=7.1Hz,5H).

[0416] Example 65: Synthesis of R244

[0417]

[0418] Step 1: In a 250 mL round-bottom flask, add compound R244-1 (2.28 g, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL), heat to 95 ° C and stir overnight in a nitrogen atmosphere. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound R244-2 (white solid, 1.20 g).

[0419] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (303 mg, 1.0 mmol, 1.2 eq), R244-2 (196 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R244-3 (white solid, 252 mg).

[0420] Step 3: Compound R244-3 (209 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45 °C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R244-4 (white solid, 163 mg).

[0421] Step 4: Compound R244-4 (81 mg, 0.2 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (column model: Spherical C18, 20-45 μM; mobile phase water: acetonitrile = 1:5) to obtain compound R244 (white solid, 49 mg). LCMS: [M+H] + =404; 1 H NMR(500MHz,DMSO-d6)δ7.81(dd,J=2.1,0.8Hz,1H),7.44–7.36(m,2H),7.20–7.14(m,1H),7.07(s,2H ),2.73–2.65(m,1H),2.63–2.53(m,6H),1.73–1.65(m,2H),1.48–1.40(m,2H),1.25(t,J=7.1Hz,6H).

[0422] Example 66: Synthesis of R247

[0423]

[0424] Step 1: In a 250 mL round-bottom flask, add compound R247-1 (2.41 g, 10.0 mmol, 1.0 eq), 3-methylpyrazole-4-boronic acid (1.89 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL), heat to 95 ° C and stir overnight in a nitrogen atmosphere. Concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound R247-2 (white solid, 1.08 g).

[0425] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (303 mg, 1.0 mmol, 1.2 eq), R247-2 (194 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R247-3 (white solid, 226 mg).

[0426] Step 3: Compound R247-3 (202 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45 °C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane: methanol=20:1) to obtain compound R247 (white solid, 166 mg). LCMS: [M+H] + =402; 1 H NMR(500MHz,DMSO-d6)δ8.26(s,1H),8.02(s,1H),7.90(dd,J=2.1,0.7Hz,1H),7.56(s,2H),7.2 9(d,J=8.6Hz,1H),7.26–7.21(m,1H),2.69–2.61(m,1H),1.73–1.65(m,2H),1.48–1.40(m,2H).

[0427] Example 67: Synthesis of R248

[0428]

[0429] Step 1: In a 250 mL round-bottom flask, add compound R244-1 (2.28 g, 10.0 mmol, 1.0 eq), 3,5-dimethylpyrazole-4-boronic acid pinacol ester (3.33 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL), heat to 95 ° C and stir overnight in a nitrogen atmosphere. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound R248-1 (white solid, 1.22 g).

[0430] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (303 mg, 1.0 mmol, 1.2 eq), R248-1 (195 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R248-2 (white solid, 233 mg).

[0431] Step 3: Compound R248-2 (209 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45 °C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R248-3 (white solid, 168 mg).

[0432] Step 4: Compound R248-3 (81 mg, 0.2 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (column model: Spherical C18, 20-45 μM; mobile phase water: acetonitrile = 1:5) to obtain compound R248 (white solid, 52 mg). LCMS: [M+H] + =403; 1 H NMR(500MHz,DMSO-d6)δ7.81(dd,J=2.1,0.8Hz,1H),7.44–7.36(m,2H),7.20–7.14(m,1H),7.07(s,2H),2.73–2. 65(m,1H),2.59(qd,J=7.1,0.9Hz,4H),2.47(s,2H),1.73–1.65(m,2H),1.48–1.40(m,2H),1.25(t,J=7.1Hz,6H).

[0433] Example 68: Synthesis of R249

[0434]

[0435] Step 1: In a 250 mL round-bottom flask, add compound R242-1 (2.14 g, 10.0 mmol, 1.0 eq), 3,5-dimethylpyrazole-4-boronic acid pinacol ester (3.33 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL), heat to 95 ° C and stir overnight in a nitrogen atmosphere. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound R249-1 (white solid, 1.18 g).

[0436] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, methyl 2-iodo-5-methylbenzoate (302 mg, 1.0 mmol, 1.2 eq), R249-1 (183 mg, 0.8 mmol), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) were added in sequence, and the reaction system was heated to 110 ° C., reacted for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R249-2 (white solid, 202 mg).

[0437] Step 3: Compound R249-2 (189 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45 °C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R249-3 (white solid, 155 mg).

[0438] Step 4: Compound R249-3 (73 mg, 0.2 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (column model: Spherical C18, 20-45 μM; mobile phase water: acetonitrile = 1:5) to obtain compound R249 (white solid, 48 mg). LCMS: [M+H] + =363; 1H NMR(500MHz,DMSO-d6)δ7.66–7.62(m,1H),7.58(s,1H),7.28–7.20(m,2H),7.19–7.14(m,1H),7.11–7.06(m,1H),6 .76(s,1H),2.59(qd,J=7.1,0.9Hz,2H),2.47(s,2H),2.42(s,2H),2.37(s,2H),2.26(s,3H),1.25(t,J=7.1Hz,3H).

[0439] Example 69: Synthesis of R250

[0440]

[0441] Step 1: In a 250 mL round-bottom flask, add compound R240-1 (2.37 g, 10.0 mmol, 1.0 eq), 3,5-dimethylpyrazole-4-boronic acid pinacol ester (3.33 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL), heat to 95 ° C and stir overnight in a nitrogen atmosphere. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound R250-1 (white solid, 1.38 g).

[0442] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, methyl 2-iodo-5-methylbenzoate (302 mg, 1.0 mmol, 1.2 eq), R250-1 (201 mg, 0.8 mmol), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) were added in sequence, and the reaction system was heated to 110 ° C., reacted for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R250-2 (white solid, 231 mg).

[0443] Step 3: Compound R250-2 (200 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45 °C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R250-3 (white solid, 186 mg).

[0444] Step 4: Compound R250-3 (77 mg, 0.2 mmol, 1.0 eq) was dissolved in N, N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (column model: Spherical C18, 20-45 μM; mobile phase water: acetonitrile = 1:5) to obtain compound R250 (white solid, 51 mg). LCMS: [M+H] + =385; 1 H NMR(500MHz,DMSO-d6)δ8.23(s,1H),7.66–7.62(m,1H),7.28–7.24(m,1H),7.16(ddd,J=7.7 ,2.5,0.9Hz,1H),7.05(d,J=2.2Hz,1H),6.74(s,1H),3.89(s,2H),2.47(s,2H),2.42(s,2H).

[0445] Example 70: Synthesis of R251

[0446]

[0447] Step 1: In a 250 mL round-bottom flask, add compound R244-1 (2.28 g, 10.0 mmol, 1.0 eq), 3,5-dimethylpyrazole-4-boronic acid pinacol ester (3.33 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL), heat to 95 ° C and stir overnight in a nitrogen atmosphere. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound R251-1 (white solid, 1.24 g).

[0448] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, 2-iodo-5-methyl-benzoic acid methyl ester (302 mg, 1.0 mmol, 1.2 eq), R251-1 (195 mg, 0.8 mmol), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) were added in sequence, and the reaction system was heated to 110 ° C., reacted for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R251-2 (white solid, 224 mg).

[0449] Step 3: Compound R251-2 (196 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45 °C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R251-3 (white solid, 155 mg).

[0450] Step 4: Compound R251-3 (76 mg, 0.2 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (column model: Spherical C18, 20-45 μM; mobile phase water: acetonitrile = 1:5) to obtain compound R251 (white solid, 45 mg). LCMS: [M+H] + =377; 1 H NMR (500MHz, DMSO-d6) δ7.66–7.62(m,1H),7.31(s,1H),7.25(d,J=7.9Hz,1H),7.19–7.14(m,1H),6. 76(s,1H),2.59(qd,J=7.1,0.9Hz,3H),2.47(s,2H),2.42(s,2H),2.37(s,2H),1.25(t,J=7.1Hz,5H).

[0451] Example 71: Synthesis of R252

[0452]

[0453] Step 1: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (303 mg, 1.0 mmol, 1.2 eq), R249-1 (183 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R252-1 (white solid, 214 mg).

[0454] Step 2: Compound R252-1 (202 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45 °C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R252-2 (white solid, 170 mg).

[0455] Step 3: Compound R252-2 (78 mg, 0.2 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (column model: Spherical C18, 20-45 μM; mobile phase water: acetonitrile = 1:5) to obtain compound R252 (white solid, 61 mg). LCMS: [M+H] + =389; 1 H NMR(500MHz,DMSO-d6)δ7.81(dd,J=2.1,0.8Hz,1H),7.68(s,1H),7.33(s,1H),7.22(dt,J=2.0,0.8Hz,1H),7.20–7.14(m,1H),7.11–7.06(m,1H),7 .07(s,2H),2.73–2.65(m,1H),2.59(qd,J=7.1,0.9Hz,2H),2.47(s,2H), 2.42(s,2H),1.73–1.65(m,2H),1.48–1.40(m,2H),1.25(t,J=7.1Hz,3H).

[0456] Example 72: Synthesis of R253

[0457]

[0458] Step 1: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (303 mg, 1.0 mmol, 1.2 eq), R250-2 (201 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain compound R253-1 (white solid, 228 mg).

[0459] Step 2: Compound R253-1 (213 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=10:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane:methanol=20:1) to obtain compound R253-2 (white solid, 190 mg).

[0460] Step 3: Compound R253-2 (82 mg, 0.2 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (2 mL), HATU (115 mg, 0.3 mmol, 1.5 eq) was added, and the mixture was stirred for 10 minutes. DIPEA (78 mg, 0.6 mmol, 3.0 eq) and ammonium chloride (13 mg, 0.24 mmol, 1.2 eq) were then added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature and separated by reverse phase column chromatography (column model: Spherical C18, 20-45 μM; mobile phase water: acetonitrile = 1:5) to obtain compound R253 (white solid, 56 mg). LCMS: [M+H] + =411; 1 H NMR(500MHz,DMSO-d6)δ8.34(s,1H),7.81(dd,J=2.1,0.7Hz,1H),7.38(s,0H),7.26(d,J=2.0Hz,1H),7.20–7.1 4(m,1H),7.09–7.03(m,2H),3.89(s,2H),2.73–2.65(m,1H),2.47(s,2H),1.73–1.65(m,2H),1.48–1.40(m,2H).

[0461] Example 73: Synthesis of R254

[0462]

[0463] Step 1: In a 250 mL round-bottom flask, add compound R254-1 (2.30 g, 10.0 mmol, 1.0 eq), 3,5-dimethylisoxazole-4-boronic acid (2.11 g, 15 mmol, 1.5 eq), Pd(dppf)Cl2 (732 mg, 1.0 mmol, 0.1 eq), potassium carbonate (2.76 g, 20.0 mmol, 2.0 eq), dioxane / water (50 mL / 5 mL), heat to 95 ° C and stir overnight in a nitrogen atmosphere. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 60: 1) to obtain compound R254-2 (white solid, 1.25 g).

[0464] Step 2: In a 50 mL round-bottom flask, under a nitrogen atmosphere, add B-3a (302 mg, 1.0 mmol), R254-2 (197 mg, 0.8 mmol, 1.0 eq), Pd(OAc)2 (18 mg, 0.08 mmol, 0.1 eq), XantPhos (70 mg, 0.12 mmol, 0.15 eq), cesium carbonate (392 mg, 1.2 mmol, 1.5 eq), and toluene (8 mL) in sequence, heat the reaction system to 110 ° C, react for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain compound R254-3 (white solid, 220 mg).

[0465] Step 3: Compound R254-3 (210 mg, 0.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran / ethanol (3 mL / 6 mL), and a solution of lithium hydroxide (60 mg, 2.5 mmol, 5.0 eq) in water (1.5 mL) was added, and the mixture was heated to 45 °C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (2 M). The solution was extracted with dichloromethane / methanol (v / v=20:1) (50 mL x 2), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane: methanol=20:1) to obtain compound R254 (white solid, 120 mg). LCMS: [M+H] + =407; 1H NMR(500MHz, DMSO-d6)δ8.34(d,J=4.0Hz,1H),7.90(dd,J=2.1,0.7Hz,1H),7.31–7.18(m,4H),3.02(pd,J= 5.6,0.7Hz,1H),2.69–2.61(m,1H),2.55(s,2H),2.02–1.93(m,2H),1.77–1.65(m,4H),1.48–1.40(m,2H).

[0466] Example 74: Synthesis of R383

[0467]

[0468] Step 1: In a 100 mL round-bottom flask, add compound R383-1 (1.0 g, 3.9 mmol), 3,5-dimethyl-1-(2-tetrahydropyranyl)-1H-pyrazole-4-boronic acid pinacol ester (1.8 g, 5.8 mmol), Pd(dppf)Cl2 (282.6 mg, 0.4 mmol), potassium carbonate (1.1 g, 7.7 mmol), dioxane / water (30 mL / 3 mL), heat to 95 ° C in a nitrogen atmosphere and stir overnight. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound R383-2 (white solid, 1.7 g).

[0469] Step 2: Under nitrogen protection, R383-2 (1.4 g, 3.9 mmol) and tetrahydrofuran (50 mL) were added to a 250 mL three-necked flask in sequence, the reaction system was cooled to -78 ° C, n-butyl lithium (2.5 M, 2.1 mL, 5.2 mmol) was added dropwise, after the addition was complete, the reaction was kept at -78 ° C for 1 hour, iodine (1.8 g, 7.0 mmol) was dissolved in tetrahydrofuran (10 mL) and added dropwise to the above reaction system. After keeping the reaction for 2 hours, the reaction was quenched with saturated sodium thiosulfate / sodium carbonate aqueous solution (10 / 1, v / v), extracted with ethyl acetate (100 mL x 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (petroleum ether: ethyl acetate = 40: 1) to obtain compound R383-3 (white solid, 1.2 g).

[0470] Step 3: In a 25 mL round-bottom flask, under a nitrogen atmosphere, R383-3 (200 mg, 0.4 mmol), B-2a (115 mg, 0.6 mmol), Pd2(dba)3 (45.8 mg, 0.04 mmol), XantPhos (43 mg, 0.08 mmol), cesium carbonate (244.5 mg, 0.8 mmol), and toluene (5 mL) were added in sequence, and the reaction system was heated to 115 ° C for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain compound R383-4 (white solid, 94.3 mg).

[0471] Step 4: Compound R383-4 (94.3 mg, 0.17 mmol) was dissolved in tetrahydrofuran / methanol (2 mL / 4 mL), and a solution of lithium hydroxide (41.4 mg, 1.70 mmol) in water (1 mL) was added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R383-4 (white solid, 80 mg).

[0472] Step 5: Compound R383-4 (50 mg, 0.09 mmolq) was dissolved in hydrochloric acid / ethyl acetate (4 M, 2 mL), and the reaction was stirred at room temperature overnight. Solid precipitated and was filtered to obtain compound R383 (hydrochloride, white solid, 25 mg). LCMS: [M+H] + =450; 1 HNMR(400MHz,DMSO-d6)δ9.39(d,J=3.7Hz,1H),7.64(d,J=2.1Hz,1H),7.49–7.34(m,1H),7.28(d,J=30.2Hz,1H), 7.14(dd,J=8.7,2.1Hz,1H),6.51(dd,J=8.6,4.4Hz,1H),2.37–2.18(m,7H),0.95–0.84(m,2H),0.61–0.57(m,2H).

[0473] Example 75: Synthesis of R375

[0474]

[0475] Step 1: In a 25 mL round-bottom flask, under a nitrogen atmosphere, add M50-1 (200 mg, 0.5 mmol), 2-amino-5-methyl-benzoic acid methyl ester (68 mg, 0.4 mmol), Pd2(dba)3 (38 mg, 0.04 mmol), XantPhos (36 mg, 0.06 mmol), cesium carbonate (204 mg, 0.6 mmol), and toluene (4 mL) in sequence, heat the reaction system to 110 ° C for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1) to obtain compound R375-1 (white solid, 247 mg).

[0476] Step 2: Dissolve compound 375-1 (54.5 mg, 0.12 mmol) in tetrahydrofuran / methanol (2 mL / 4 mL), add lithium hydroxide (14.4 mg, 0.60 mmol) in water (1 mL), heat the reaction to 45 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 6-7 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R375 (white solid, 87 mg). LCMS: [M+H] + =425; 1 HNMR(400MHz,DMSO-d6)δ13.26(s,1H),9.52(s,1H),7.73(s,1H),7.55(dd,J=11.2,1.5Hz,1H),7.43(s ,1H),7.25(dd,J=8.5,1.5Hz,1H),6.53(dd,J=8.4,4.6Hz,1H),2.47(s,3H),2.30(s,3H),2.24(s,3H).

[0477] Example 76: Synthesis of R377

[0478]

[0479] Step 1: In a 25 mL round-bottom flask, R383-3 (200 mg, 0.41 mmol), methyl 2-amino-5-methylbenzoate (56.0 mg, 0.3 mmol), Pd2(dba)3 (32 mg, 0.03 mmol), XantPhos (30 mg, 0.05 mmol), cesium carbonate (168 mg, 0.5 mmol), and toluene (5 mL) were added in sequence under a nitrogen atmosphere, and the reaction system was heated to 115 ° C for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain compound R377-1 (white solid, 33.5 mg).

[0480] Step 2: In a 25 mL round-bottom flask, compound R377-1 (97.5 mg, 0.06 mmol) was dissolved in tetrahydrofuran / methanol (2 mL / 4 mL), and a solution of lithium hydroxide (15.4 mg, 0.6 mmol) in water (1 mL) was added, and the reaction was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R377-2 (white solid, 35 mg).

[0481] Step 5: Dissolve compound R377-2 (35 mg, 0.06 mmol) in hydrochloric acid / ethyl acetate (4 M, 2 mL) and stir overnight at room temperature. Solid precipitates and is filtered to obtain compound R377 (hydrochloride salt, white solid, 20 mg). LCMS: [M+H] + =424; 1 HNMR(400MHz,DMSO-d6)δ9.40(s,1H),7.72(s,1H),7.49–7.36(m,1H),7.28(s,1H), 7.24(dd,J=8.6,1.8Hz,1H),6.51(dd,J=8.5,4.3Hz,1H),2.29(s,6H),2.24(s,3H).

[0482] Example 77: Synthesis of R385

[0483]

[0484] In a 25mL round-bottom flask, compound R383-5 (30mg, 0.06mmol) was dissolved in toluene (2mL), and thionyl chloride (28.6mg, 0.24mmol) was slowly added. The reaction system was heated to 100℃ and stirred for 50 minutes. After concentration to dryness, the crude product was dissolved in dichloromethane (2mL), and added dropwise to a 25mL single-mouth bottle containing ammonia water (1mL) at 0℃, and the temperature was naturally raised to room temperature and stirred overnight. After concentration to dryness under reduced pressure, the crude product was separated by column chromatography (dichloromethane: methanol = 20:1) to obtain compound R385 (white solid, 25mg). LCMS: [M+H] + =449; 1HNMR (400MHz, DMSO-d6) δ12.44(s,1H),9.94(s,1H),8.13(s,1H),7.56–7.38(m,2H),7.37–7.26(m,1H),7.21(s,1H),7.07(dd,J=8.5,1. 7Hz,1H),6.46(dd,J=8.5,4.6Hz,1H),2.30(d,J=31.3Hz,6H),1.83(ddd,J=13.5,8.7,5.2Hz,1H),0.93–0.81(m,2H),0.71–0.62(m,2H).

[0485] Example 78: Synthesis of R334

[0486]

[0487] Step 1: In a 100 mL round-bottom flask, under a nitrogen atmosphere, M45-1 (100 mg, 0.3 mmol), methyl 2-amino-5-methoxybenzoate (52 mg, 0.3 mmol), Pd(OAc)2 (10 mg, 0.03 mmol), XantPhos (23 mg, 0.04 mmol), cesium carbonate (128 mg, 0.4 mmol), and toluene (4 mL) were added in sequence, and the reaction system was heated to 110 ° C for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound R334-1 (yellow oil 50 mg).

[0488] Step 2: Compound R334-1 (50 mg, 0.11 mmol) was dissolved in tetrahydrofuran / methanol (1 mL / 2 mL), and a solution of lithium hydroxide (13.7 mg, 0.57 mmol) in water (1 mL) was added. The reaction was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH 6-7 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R334 (white solid, 33 mg). LCMS: [M+H] + =423; 1 HNMR (400MHz, DMSO) δ13.30(s,1H),9.06(d,J=15.8Hz,1H),7.47(s,1H),7.41(d,J=3.1Hz,1H),7.36(dd,J=11.2,1.7Hz ,1H),7.29(s,1H),7.20(s,1H),7.11–7.04(m,1H),6.57(dd,J=9.1,4.3Hz,1H),3.73(s,3H),2.47(s,3H),2.29(s,3H).

[0489] Example 79: Synthesis of R335

[0490]

[0491] Step 1: In a 100 mL round-bottom flask, under a nitrogen atmosphere, add M45-1 (100 mg, 0.3 mmol), methyl 2-amino-5-methylbenzoate (48 mg, 0.3 mmol), Pd(OAc)2 (10 mg, 0.03 mmol), XantPhos (23 mg, 0.04 mmol), cesium carbonate (128 mg, 0.4 mmol), and toluene (4 mL) in sequence, heat the reaction system to 110 ° C for 24 hours, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound R335-1 (yellow oil, 52 mg).

[0492] Step 2: Compound R335-1 (42 mg, 0.1 mmol) was dissolved in tetrahydrofuran / methanol (1 mL / 2 mL), and a solution of lithium hydroxide (14 mg, 0.6 mmol) in water (1 mL) was added. The reaction was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH 6-7 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R335 (white solid, 32 mg). LCMS: [M+H] + =407; 1 HNMR (400MHz, DMSO-d6) δ9.37 (s, 1H), 7.72 (d, J = 1.7Hz, 1H), 7.47, 7.29, 7.10 (m, 1H), 7.37 (dd, J = 11. 1,1.8Hz,1H),7.25–7.17(m,2H),6.49(dd,J=8.5,4.3Hz,1H),2.48(s,3H),2.30(s,3H),2.24(s,3H).

[0493] Example 80: Synthesis of R340

[0494]

[0495] Step 1: In a 100 mL round-bottom flask, add compound R340-1 (441 mg, 2.0 mmol), 3,5-dimethyl-1-(2-tetrahydropyranyl)-1H-pyrazole-4-boronic acid pinacol ester (796 mg, 2.6 mmol), Pd(dppf)Cl2 (47 mg, 0.2 mmol), potassium carbonate (552 mg, 4.0 mmol), dioxane / water (20 mL / 2 mL), heat to 95 ° C in a nitrogen atmosphere and stir overnight. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain compound R340-2 (white solid, 334 mg).

[0496] Step 2: In a 25 mL round-bottom flask, under a nitrogen atmosphere, R340-2 (334 mg, 1.0 mmol), methyl 2-iodo-5-methylbenzoate (355 mg, 1.3 mmol), Pd2(dba)3 (95.7 mg, 0.10 mmol), XantPhos (90.7 mg, 0.16 mmol), cesium carbonate (515 mg, 1.6 mmol), and toluene (10 mL) were added in sequence, and the reaction system was heated to 115 ° C for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 30:1) to obtain compound R340-3 (white solid, 396 mg).

[0497] Step 3: Compound R340-3 (390 mg, 0.8 mmol) was dissolved in tetrahydrofuran / methanol (5 mL / 10 mL), and a solution of lithium hydroxide (100 mg, 4.2 mmol) in water (2 mL) was added, and the mixture was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R340-4 (white solid, 157 mg).

[0498] Step 4: In a 25mL round-bottom flask, compound R340-4 (125mg, 0.3mmol) was dissolved in toluene (2mL), and dichlorothionyl (130mg, 1.2mmol) was slowly added, and the reaction system was heated to 100°C and stirred for 50 minutes. The reaction solution was concentrated to dryness, and the crude product was dissolved in 6mL of dichloromethane and added dropwise to a 25mL single-mouth bottle containing 3mL of ammonia water at 0°C, and the temperature was naturally raised to room temperature and stirred overnight, and the mixture was concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20:1) to obtain compound R340 (white solid, 25mg). LCMS: [M+H] + =369; 1HNMR(400MHz,DMSO-d6)δ12.31(s,1H),9.65(s,1H),8.00(s,1H),7.56(s,1H),7.36(s,1H),7.28(s,1 H),7.21(s,1H),7.05(d,J=8.4Hz,1H),6.13(d,J=8.3Hz,1H),2.23(s,6H),2.21(s,3H),2.19(s,3H).

[0499] Example 81: Synthesis of R308

[0500]

[0501] Step 1: In a 100 mL round-bottom flask, add compound R308-1 (1.0 g, 5.0 mmol), N-chlorosuccinimide (665 mg, 5.0 mmol), N,N-dimethylformamide (20 mL), and heat the reaction to 70 ° C and stir overnight. Pour the reaction solution into water (150 mL), extract with ethyl acetate (50 mL x 3), combine the organic phases, wash with saturated brine (20 mL), dry with anhydrous sodium sulfate, filter, and concentrate. The crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound R308-2 (white solid, 1.2 g).

[0502] Step 2: In a 100 mL round-bottom flask, add compound R308-2 (1.2 g, 5.0 mmol), 3,5-dimethyl-1-(2-tetrahydropyranyl)-1H-pyrazole-4-boronic acid pinacol ester (2.3 g, 7.5 mmol), Pd(dppf)Cl2 (366 mg, 0.5 mmol), potassium carbonate (1.4 g, 10 mmol), dioxane / water (20 mL / 2 mL), heat to 95 ° C in a nitrogen atmosphere and stir overnight. Concentrate, and the crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain compound R308-3 (white solid, 1.1 g).

[0503] Step 3: In a 25 mL round-bottom flask, under a nitrogen atmosphere, R308-3 (100 mg, 0.30 mmol), methyl 2-iodo-5-methylbenzoate (105 mg, 0.4 mmol), Pd2(dba)3 (55 mg, 0.06 mmol), XantPhos (52 mg, 0.09 mmol), cesium carbonate (145 mg, 0.5 mmol), and toluene (5 mL) were added in sequence, and the reaction system was heated to 115 ° C for 24 hours, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain compound R308-4 (white solid, 124 mg).

[0504] Step 4: Compound R308-4 (123.6 mg, 0.26 mmol) was dissolved in tetrahydrofuran / methanol (5 mL / 10 mL), and a solution of lithium hydroxide (63 mg, 2.6 mmol) in water (1 mL) was added, and the reaction was heated to 45°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R308-4 (white solid, 111 mg).

[0505] Step 5: In a 25 mL round-bottom flask, compound R308-4 (50.0 mg, 0.11 mmol) was dissolved in 4 mL of toluene, and thionyl chloride (50.8 mg, 0.43 mmol) was slowly added, and the reaction system was heated to 100°C and stirred for 50 minutes. Concentrate to dryness, dissolve the crude product in 6 mL of dichloromethane, and add dropwise to a 25 mL single-mouth bottle containing 3 mL of ammonia water at 0°C, naturally warm to room temperature and stir overnight, and concentrate to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 40: 1) to obtain compound R308 (white solid, 18 mg). LCMS: [M+H] + =383; 1 HNMR(600MHz,DMSO-d6)δ12.36(s,1H),9.70(s,1H),7.99(s,1H),7.55(s,1H),7.35(s,1H),7.29(d,J=1.9Hz,1H),7.21(d,J=1.7H z,1H),7.04(dd,J=8.4,1.4Hz,1H),6.12(d,J=8.4Hz,1H),2.55(m,2H),2.26(d,J=26.5Hz,6H),2.20(s,3H),1.09(t,J=7.5Hz,3H).

[0506] Example 82: Synthesis of R431

[0507]

[0508] Step 1: In a 25 mL round-bottom flask, add R86 (50.0 mg, 0.13 mmol), toluene (5 mL), and thionyl chloride (0.5 mL) in sequence, heat the reaction system to 100 °C and stir for 0.5 hours. Concentrate to dryness under reduced pressure to obtain compound R431-1 (yellow solid, crude product), which is used directly in the next step.

[0509] Step 2: In a 25 mL round-bottom flask, add dichloromethane (2 mL), methylamine hydrochloride (17.4 mg, 0.26 mmol) and triethylamine (51.0 mg, 0.5 mmol) in sequence, then add a dichloromethane (2 mL) solution of the crude product of compound R431-1 to the above system, and stir the reaction at 25°C overnight. Adjust the pH to 5-6 with dilute hydrochloric acid, and concentrate to dryness under reduced pressure. The crude product is separated by column chromatography (dichloromethane: methanol = 40: 1) to obtain compound R431 (white solid, 23.4 mg). LCMS: [M+H] + =394; 1 HNMR (400MHz, CDCl3) δ8.99(s,1H),7.19(s,1H),7.00(dd,J=8.6,2.1Hz,1H),6.94(s,1H),6.90(d,J=10.9Hz,1H),6.49(dd,J=8.6, 3.6Hz,1H),6.21(s,1H),3.05(d,J=4.8Hz,3H),2.46(s,3H),2.34(s,3H),1.98-1.85(m,1H),0.94–0.89(m,3H),0.68-0.62(m,2H).

[0510] Example 83: Synthesis of R434

[0511]

[0512] In a 25mL round-bottom flask, dichloromethane (2mL), cyclobutylamine (18.6mg, 0.26mmol) and triethylamine (51.0mg, 0.5mmol) were added in sequence, and then the dichloromethane (2mL) solution of compound R431-1 was added to the above system, and the reaction was stirred at 25°C overnight. The pH was adjusted to 5-6 with dilute hydrochloric acid, and the mixture was concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 40: 1) to obtain compound R434 (white solid, 9.4mg). LCMS: [M+H] + =434; 1 H NMR (400MHz, CDCl3) δ8.99(s,1H),7.20(s,1H),6.98(d,J=8.6Hz,1H),6.93(s,1H),6.89(d,J=10.5Hz,1H),6.48(s,1H),6 .29(s,1H),4.60(s,1H),2.47(d,J=10.8Hz,4H),2.33(d,J=5.7Hz,6H),2.04(s,2H),0.92-0.88(m,2H),0.66-0.63(m,2H).

[0513] Example 84: Synthesis of R422

[0514]

[0515] Step 1: In a 100 mL round-bottom flask, compound R422-1 (1.0 g, 4.1 mmol), 3,5-dimethyl-1-(2-tetrahydropyranyl)-1H-pyrazole-4-boronic acid pinacol ester (2.0 g, 6.5 mmol), Pd(dppf)Cl2·CH2Cl2 (175 mg, 0.2 mmol), sodium carbonate (1.1 g, 10.6 mmol), DMSO / water (20 mL / 2.4 mL) were added in sequence, and heated to 80°C in a nitrogen atmosphere and stirred overnight. The reaction solution was diluted with ethyl acetate (200 mL), washed with saturated brine (100 mL x 3), the organic phase was dried over anhydrous sodium sulfate and concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R422-2 (white solid, 780 mg).

[0516] Step 2: In a 25 mL round-bottom flask, R422-2 (530.5 mg, 1.56 mmol), methyl 2-iodo-5-methylbenzoate (516 mg, 1.87 mmol), XantPhos (135 mg, 0.23 mmol), Cs2CO3 (763 mg, 2.34 mmol), toluene (8 mL), Pd2(dba)3 (143 mg, 0.15 mmol) were added in sequence under nitrogen atmosphere, and the reaction system was heated to 110°C and stirred for 24 hours. The reaction solution was concentrated to dryness, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain compound R422-3 (light yellow solid, 738.7 mg).

[0517] Step 3: Weigh compound R422-3 (738.7 mg, 1.5 mmol) in tetrahydrofuran / methanol (6 mL / 12 mL), add lithium hydroxide (218 mg, 9.1 mmol) in water (5 mL), heat the reaction to 50 ° C and stir overnight. The reaction solution is cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), concentrated to dryness under reduced pressure, and the crude product is separated by column chromatography (dichloromethane: methanol = 30: 1) to obtain compound R422-4 (light yellow solid, 494.2 mg).

[0518] Step 4: In a 25mL round-bottom flask, compound R422-4 (91mg, 0.2mmol) was dissolved in 2mL of toluene, and dichlorothionyl (98.4mg, 0.8mmol) was slowly added, and the reaction system was heated to 100°C and stirred for 50 minutes. The reaction was concentrated to dryness, and the crude product was dissolved in dichloromethane (2mL), and added dropwise to a 25mL single-mouth bottle containing ammonia water (1mL) at 0°C, and the temperature was naturally raised to room temperature and stirred overnight, and the mixture was concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20:1) to obtain compound R422 (white solid, 37.5mg). LCMS: [M+H] + =389; 1 H NMR 1 H NMR (400MHz, DMSO-d6) δ12.44(s,1H),9.88(s,1H),8.04(s,1H),7.56(d,J=2.0Hz, 1H),7.45(s,3H),7.09(dd,J=8.5,2.0Hz,1H),6.23(d,J=8.4Hz,1H),2.23(s,3H).

[0519] Example 85: Synthesis of R426

[0520]

[0521] Step 1: In a 25 mL round-bottom flask, R340-2 (260 mg, 0.8 mmol), B-3a (319 mg, 1.1 mmol), XantPhos (69.5 mg, 0.12 mmol), Cs2CO3 (396 mg, 1.2 mmol), toluene (5 mL), Pd2(dba)3 (74.2 mg, 0.1 mmol) were added in sequence under nitrogen atmosphere, and the reaction system was heated to 110°C and stirred for 24 hours. The reaction solution was concentrated to dryness, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R426-1 (off-white solid, 433 mg).

[0522] Step 2: Compound R426-1 (430 mg, 0.8 mmol) was dissolved in tetrahydrofuran / methanol (5 mL / 10 mL), and a solution of lithium hydroxide (126 mg, 5.3 mmol) in water (5 mL) was added, and the reaction was heated to 50°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound R426-2 (light yellow solid, 285 mg).

[0523] Step 3: In a 25 mL round-bottom flask, compound R426-2 (285 mg, 0.6 mmol) was dissolved in 5 mL of toluene, and dichlorothionyl (285 mg, 2.4 mmol) was added, and the reaction was heated to 100°C and stirred for 50 minutes. After the solvent was dried, it was dissolved in 5 mL of dichloromethane and added dropwise to a 25 mL single-mouth bottle containing 5 mL of ammonia water at 0°C, and the temperature was naturally raised to room temperature and stirred overnight, and the mixture was concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20:1) to obtain compound R426 (white solid, 101 mg). LCMS: [M+H] + =395; 1 H NMR (400MHz, DMSO-d6) δ12.35(s,1H),9.64(s,1H),8.05(s,1H),7.43–7.33(m,2H),7.28(d,J=2.0Hz,1H),7.20(d,J=2.0Hz,1H),6. 99(dd,J=8.5,2.1Hz,1H),6.11(d,J=8.5Hz,1H),2.23(s,5H),2.18(s,3H),1.89–1.80(m,1H),0.92–0.85(m,2H),0.69–0.60(m,2H).

[0524] Example 86: Synthesis of R427

[0525]

[0526] Step 1: In a 100 mL round-bottom flask, compound R422-1 (1.0 g, 4.1 mmol), R427-1 (1.7 g, 6.5 mmol), Pd(dppf)Cl2·CH2Cl2 (175 mg, 0.2 mmol), sodium carbonate (1.1 g, 10.7 mmol), DMSO / water (20 mL / 2.4 mL) were added in sequence, and heated to 80°C in a nitrogen atmosphere and stirred overnight. The reaction solution was diluted with ethyl acetate (300 mL), washed with saturated brine (100 mL x 3), the organic phase was dried over anhydrous sodium sulfate and concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R427-2 (white solid, 980 mg).

[0527] Step 2: In a 25 mL round-bottom flask, R427-2 (275 mg, 0.8 mmol), B-3a (350 mg, 1.2 mmol), XantPhos (67 mg, 0.12 mmol), Cs2CO3 (380 mg, 1.2 mmol), toluene (5 mL), Pd2(dba)3 (71 mg, 0.1 mmol) were added in sequence under nitrogen atmosphere, and the reaction system was heated to 110 ° C and stirred for 24 hours. The reaction solution was concentrated to dryness, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain compound R427-3 (yellow oil, 256 mg).

[0528] Step 3: Compound R427-3 (256 mg, 0.5 mmol) was dissolved in tetrahydrofuran / methanol (5 mL / 10 mL), and a solution of lithium hydroxide (72 mg, 3.0 mmol) in water (5 mL) was added, and the reaction was heated to 50°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure to obtain crude compound R427-4 (yellow oil 189 mg).

[0529] Step 4: In a 25mL round-bottom flask, compound R427-4 (190mg, 0.4mmol) was dissolved in toluene (5mL), and dichlorothionyl (180mg, 1.5mmol) was slowly added. The reaction system was heated to 100°C and stirred for 50 minutes. The mixture was concentrated to dryness under reduced pressure, and the crude product was dissolved in dichloromethane (5mL), and added dropwise to a 25mL single-mouth bottle containing ammonia water (5mL) at 0°C, and the temperature was naturally raised to room temperature and stirred overnight. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane: methanol = 20:1) to obtain compound R427 (white solid, 64.9mg). LCMS: [M+H] + =415; 1 H NMR (400MHz, DMSO-d6) δ12.44(s,1H),9.88(d,J=9.0Hz,1H),8.10(s,1H),7.54–7.35(m,3H),7.03(s,1H) ,6.22(t,J=9.0Hz,1H),2.25(s,4H),1.82(brs,1H),0.86–0.82(s,2H),0.66–0.62(q,J=7.4,6.0Hz,2H).

[0530] Example 87: Synthesis of R501

[0531]

[0532] Step 1: Compound R308-3 (167 mg, 0.5 mmol) was dissolved in toluene (2.0 mL), and B-3a (227 mg, 0.75 mmol), Pd2(dba)3 (91 mg, 0.1 mmol), XantPhos (116 mg, 0.2 mmol), and Cs2CO3 (250 mg, 0.8 mmol) were added in sequence. The reaction system was heated to 115°C and stirred for 20 hours under a nitrogen atmosphere. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R501-1 (yellow solid, 200 mg).

[0533] Step 2: Compound R501-1 (200 mg, 0.49 mmol) was dissolved in tetrahydrofuran / methanol / water (2 mL / 2 mL / 1 mL), lithium hydroxide (92 mg, 4.0 mmol) was added, and the reaction was heated to 50°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid, and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R501-2 (white solid, 120 mg).

[0534] Step 3: Compound R501-2 (120 mg, 0.24 mmol) was dissolved in toluene (4 mL), and thionyl chloride (0.5 mL) was slowly added. The reaction was heated to 100 ° C and stirred for 45 minutes. The mixture was concentrated to dryness under reduced pressure. The crude product was dissolved in dichloromethane (2 mL), added to ammonia water (1 mL) at 0 ° C, and naturally heated to room temperature and stirred for 3 hours. The organic phase was separated, and the aqueous phase was extracted with dichloromethane (5 mL x 2). The combined dichloromethane phases were dried over anhydrous sodium sulfate and concentrated. The crude product was separated by preparative TLC (dichloromethane: methanol = 20: 1) to obtain compound R501 (yellow solid, 27 mg). LCMS: [M+H] + =409; 1 H NMR (600MHz, DMSO-d6) δ12.34(s,1H),9.69(s,1H),8.05(s,1H),7.45–7.31(m,2H),7.29(d,J=2.0Hz,1H),7.21(d,J=2.0Hz,1H),6.98( d,J=8.4Hz,1H),6.11(d,J=8.4Hz,1H),2.57–2.53(m,2H),2.50(s,6H),1.24(s,1H),1.11(m,3H),0.92–0.84(m,2H),0.72–0.58(m,2H).

[0535] Example 88: Synthesis of R502

[0536]

[0537] Step 1: In a 100 mL round-bottom flask, compound R502-1 (2.2 g, 10 mmol), 3,5-dimethyl-1-(2-tetrahydropyranyl)-1H-pyrazole-4-boronic acid pinacol ester (1.7 g, 12 mmol), Pd(PPh3)4 (557 mg, 0.5 mmol), potassium carbonate (4.14 g, 30 mmol), toluene / water / ethanol (25 mL / 5 mL / 5 mL), heated to 100 ° C in a nitrogen atmosphere and stirred overnight. The reaction solution was diluted with ethyl acetate (200 mL), washed with saturated brine (50 mL x 3), the organic phase was dried over anhydrous sodium sulfate and concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R502-2 (white solid, 1780 mg).

[0538] Step 2: In a 100 mL round-bottom flask, add R502-2 (480 mg, 1.5 mmol), sodium thiomethoxide (315 mg, 4.5 mmol), and methanol (10 mL) in sequence. The reaction was stirred at room temperature for 3 hours and then concentrated. The crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R502-3 (white solid, 300 mg).

[0539] Step 3: In a 50 mL round-bottom flask, compound R502-3 (300 mg, 0.75 mmol), water / ethanol (2 mL / 10 mL), ammonium chloride (200 mg, 3.75 mmol), iron powder (208 mg, 3.75 mmol) were added in sequence, and the reaction was heated to 80 ° C and stirred for 6 hours. The reaction solution was concentrated to dryness, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound R502-4 (white solid, 210 mg).

[0540] Step 4: Compound R502-4 (120 mg, 0.32 mmol) was dissolved in toluene (2 mL), and methyl 2-iodo-5-methylbenzoate (176 mg, 0.64 mmol), Pd2(dba)3 (59 mg, 0.064 mmol), XantPhos (74 mg, 0.128 mmol), and Cs2CO3 (156 mg, 0.48 mmol) were added in sequence. The reaction system was heated to 115°C and stirred for 20 hours under a nitrogen atmosphere. The reaction solution was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R502-5 (yellow solid, 160 mg).

[0541] Step 5: Weigh compound R502-5 (160 mg, 0.32 mmol) in tetrahydrofuran / methanol / water (2 mL / 2 mL / 1 mL), add lithium hydroxide (61 mg, 2.56 mmol), heat the reaction to 50°C and stir overnight. The reaction solution was cooled to room temperature, adjusted to pH 5-6 with dilute hydrochloric acid (2 M), concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R502-6 (white solid, 80 mg).

[0542] Step 6: Weigh compound R502-6 (80 mg, 0.24 mmol) in toluene (4 mL), add dichlorothionyl (0.5 mL), heat the reaction to 100 ° C and stir for 45 minutes, concentrate to dryness under reduced pressure, dissolve the crude product in dichloromethane (2 mL), add ammonia water (1 mL) at 0 ° C, then naturally warm to room temperature and stir for 3 hours, separate the organic phase, extract the aqueous phase with dichloromethane (5 mL x 2), dry the combined dichloromethane phases over anhydrous sodium sulfate and concentrate, separate the crude product by preparative TLC (dichloromethane: methanol = 20: 1), and obtain compound R502 (yellow solid, 33 mg). LCMS: [M+H] + =401; 1 H NMR (600MHz, DMSO-d6) δ12.39(s,1H),9.69(s,1H),8.03–7.93(m,1H),7.55(d,J=2.1Hz,1H),7.40–7.29(m,1H),7.22(d,J=1 .8Hz,1H),7.07(d,J=1.9Hz,1H),7.03(dd,J=8.4,2.0Hz,1H),6.13(d,J=8.4Hz,1H),2.40(s,3H),2.26(s,6H),2.21(s,3H).

[0543] Example 89: Synthesis of R503

[0544]

[0545] Step 1: In a 100 mL round-bottom flask, compound R503-1 (2.37 g, 10 mmol), 3,5-dimethyl-1-(2-tetrahydropyranyl)-1H-pyrazole-4-boronic acid pinacol ester (1.7 g, 12 mmol), Pd(PPh3)4 (557 mg, 0.5 mmol), potassium carbonate (4.14 g, 30 mmol), toluene / water / ethanol (25 mL / 5 mL / 5 mL), heated to 100 ° C in a nitrogen atmosphere and stirred overnight. The reaction solution was diluted with ethyl acetate (200 mL), washed with saturated brine (50 mL x 3), the organic phase was dried over anhydrous sodium sulfate and concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R503-2 (white solid, 1680 mg).

[0546] Step 2: In a 100 mL round-bottom flask, add R503-2 (211 mg, 0.56 mmol), sodium hydroxide (2.74 g, 18 mmol), DMSO / water (2.5 mL / 0.5 mL) in sequence, and stir the reaction system for 2 hours. The reaction solution was concentrated to dryness under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R503-3 (white solid, 160 mg).

[0547] Step 3: In a 100 mL round-bottom flask, add R503-3 (2.4 g, 7.2 mmol), sodium difluorochloroacetate (56 mg, 1.4 mmol), potassium carbonate (1.98 g, 14.4 mmol), N,N-dimethylformamide / water (45 mL / 6 mL) in sequence, and heat the reaction system to 100 ° C and stir for 2 hours. Concentrate to dryness under reduced pressure, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R503-4 (yellow solid, 1.8 g).

[0548] Step 4: In a 50 mL round-bottom flask, compound R503-4 (251 mg, 0.75 mmol), water / ethanol (2 mL / 10 mL), ammonium chloride (200 mg, 3.75 mmol), iron powder (208 mg, 3.75 mmol) were added in sequence, and the reaction was heated to 80 ° C and stirred for 6 hours. The reaction solution was concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound R503-5 (white solid, 189 mg).

[0549] Step 5: Compound R503-5 (113 mg, 0.32 mmol) was weighed into toluene (2 mL), and methyl 2-iodo-5-methylbenzoate (176 mg, 0.64 mmol), Pd2(dba)3 (59 mg, 0.064 mmol), XantPhos (74 mg, 0.128 mmol), and Cs2CO3 (156 mg, 0.48 mmol) were added in sequence, and the mixture was heated to 115°C and stirred for 20 hours under a nitrogen atmosphere. The reaction solution was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R503-6 (yellow solid, 140 mg).

[0550] Step 6: Compound R503-6 (160 mg, 0.32 mmol, 1.0 eq) was dissolved in tetrahydrofuran / methanol / water (2 mL / 2 mL / 1 mL), lithium hydroxide (61 mg, 2.56 mmol) was added, and the reaction was heated to 50°C and stirred overnight. The reaction solution was cooled to room temperature, and the pH was adjusted to 5-6 with dilute hydrochloric acid (2 M). The crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain R503-7 (110 mg).

[0551] Step 7: The crude product of R503-7 (50 mg) was added to HCl / dioxane (4 M, 5 mL), stirred at 25°C for 12 hours, and filtered to obtain compound R503 (hydrochloride salt, white solid, 20 mg). LCMS: [M+H] + =406.

[0552] Example 90: Synthesis of R504

[0553]

[0554] Step 1: Compound R503-5 (113 mg, 0.32 mmol) was dissolved in toluene (2 mL), and B-3a (186 mg, 0.64 mmol), Pd2(dba)3 (59 mg, 0.064 mmol), XantPhos (74 mg, 0.128 mmol), and Cs2CO3 (156 mg, 0.48 mmol) were added in sequence, and the reaction system was heated to 115°C and stirred for 20 hours under a nitrogen atmosphere. The mixture was concentrated to dryness under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound R504-1 (yellow solid, 140 mg), which was directly used in the next step.

[0555] Step 2: Compound R504-1 (160 mg, 0.32 mmol) was dissolved in tetrahydrofuran / methanol / water (2 mL / 2 mL / 1 mL), lithium hydroxide (61 mg, 2.56 mmol) was added, and the reaction was heated to 50°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH = 5-6 with dilute hydrochloric acid (2M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R504-2 (white solid, 80 mg).

[0556] Step 3: R504-2 (30 mg) was added to HCl / dioxane (4 M, 3 mL), and the reaction was stirred at 25°C for 12 hours. Solid precipitated and was filtered to obtain compound R504 (hydrochloride salt, white solid, 10 mg). LCMS: [M+H] + =432; 1 H NMR(600MHz,DMSO--d6)δ12.41(s,1H),9.93(s,1H),8.05(s,1H),7.77–7.69(m,1H),7.31(d,J=2.1Hz,1H),7.27– 7.17(m,1H),6.68(t,J=7.5Hz,1H),6.20(d,J=8.3Hz,1H),2.57(d,J=7.4Hz,1H),2.25(s,6H),1.20–1.01(m,4H).

[0557] Example 91: Synthesis of R505

[0558]

[0559] Compound R504-2 (103 mg, 0.24 mmol) was dissolved in toluene (4 mL), and thionyl chloride (0.5 mL) was slowly added. The reaction was heated to 100 ° C and stirred for 45 minutes. The reaction solution was cooled to room temperature and concentrated to dryness under reduced pressure. The crude product was dissolved in dichloromethane (1 mL), added to ammonia water (1 mL) at 0 ° C, and naturally heated to room temperature and stirred for 3 hours. The organic phase was separated, and the aqueous phase was extracted with dichloromethane (5 mL x 2). The combined dichloromethane phases were dried over anhydrous sodium sulfate and concentrated. The crude product was separated by preparative TLC (dichloromethane: methanol = 20: 1) to obtain compound R505 (white solid, 18 mg). LCMS: [M+H] + =431.

[0560] Example 92: Synthesis of R506

[0561]

[0562] Compound R503-7 (97 mg, 0.24 mmol) was dissolved in toluene (4 mL), thionyl chloride (0.5 mL) was slowly added, and the mixture was heated to 100 °C and stirred for 45 minutes. The reaction solution was cooled to room temperature and concentrated to dryness under reduced pressure. The crude product was dissolved in dichloromethane (1 mL), added to ammonia water (1 mL) at 0 °C, and stirred for 3 hours after the temperature was naturally raised to room temperature. The organic phase was separated, and the aqueous phase was extracted with dichloromethane (5 mL x 2). The combined dichloromethane phases were dried over anhydrous sodium sulfate and concentrated. The crude product was separated by preparative TLC (dichloromethane: methanol = 20: 1) to obtain compound R506 (white solid, 29 mg). LCMS: [M+H] + =405; 1 H NMR(600MHz,DMSO-d6)δ12.41(s,1H),9.66(s,1H),8.02(s,1H),7.54(s,1H),7.38(d,J=21.6Hz,1H) ,7.24(s,1H),7.20–7.06(m,2H),7.03(s,1H),6.44(dd,J=8.5,4.0Hz,1H),2.26(s,6H),2.23(s,3H).

[0563] Example 93: Synthesis of R551

[0564]

[0565] Step 1: In a 100 mL round-bottom flask, add R551-1 (1.5 g, 7.3 mmol), N-iodosuccinimide (2.5 g, 11.0 mmol), N,N-dimethylformamide (30 mL) in sequence, and heat the reaction system to 70 ° C and stir overnight. Add water (100 mL) to the reaction system, extract with dichloromethane (50 mL x 3), combine the organic phases, wash with water (50 mL x 3) and saturated brine (50 mL) in sequence, dry over anhydrous sodium sulfate and concentrate, and separate the crude product by silica gel column chromatography (100% petroleum ether) to obtain compound R551-2 (white solid, 800 mg).

[0566] Step 2: In a 50 mL round-bottom flask, compound R551-2 (800 mg, 2.4 mmol), 3,5-dimethyl-1-(2-tetrahydropyranyl)-1H-pyrazole-4-boronic acid pinacol ester (705 mg, 2.3 mmol), Pd(dppf)Cl2 (8 mg, 0.01 mmol), potassium carbonate (954 mg, 6.9 mmol), dioxane / water (15 mL / 3 mL) were added in sequence, and heated to 80°C in a nitrogen atmosphere and stirred overnight. The reaction solution was concentrated to dryness under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound R551-3 (yellow solid, 490 mg).

[0567] Step 3: In a 50 mL round-bottom flask, R551-3 (200 mg, 0.5 mmol), methyl 2-iodo-5-methylbenzoate (288 mg, 1.1 mmol), Pd2(dba)3 (40 mg, 0.04 mmol), XantPhos (80 mg, 0.1 mmol), Cs2CO3 (256 mg, 0.8 mmol), and toluene (15 mL) were added in sequence under nitrogen atmosphere, and the reaction system was heated to 115°C and stirred overnight. The reaction solution was concentrated to dryness under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound R551-4 (yellow solid, 118 mg).

[0568] Step 4: Compound R551-4 (118 mg, 0.22 mmol) was dissolved in methanol / water (2 mL / 1 mL), lithium hydroxide (80 mg, 3.33 mmol) was added, and the mixture was heated to 50°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20: 1) to obtain compound R551-5 (white solid, 110 mg).

[0569] Step 5: In a 25 mL round-bottom flask, compound R551-5 (110 mg, 0.2 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (122 mg, 0.32 mmol) and N,N-dimethylformamide (10 mL), diisopropylethylamine (83 mg, 0.64 mmol), ammonium chloride (28 mg, 0.53 mmol) were added in sequence, the reaction system was heated to 45 ° C and stirred for 16 hours, 20 mL of water was added to the reaction system, and dichloromethane was extracted (10 mL×3), the organic phases were combined, washed with water (50 mL x 3) and saturated brine (50 mL) in sequence, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product R551-6 (yellow solid, 170 mg), which was directly used in the next step.

[0570] Step 6: Dissolve compound R551-6 (100 mg) in dioxane (3 mL), slowly add HCl / dioxane solution (4 M, 1 mL), stir the reaction at room temperature for 24 hours to precipitate solid, filter, rinse with dioxane, and dry to obtain product compound R551 (hydrochloride, white solid, 40 mg). LCMS: [M+H] + =433; 1 H NMR (600MHz, DMSO-d6) δ9.93(s,1H),8.05(s,1H),7.69(d,J=2.0Hz,1H),7.58(d,J=2.0Hz,2H ),7.42(s,1H),7.08(dd,J=8.4,2.0Hz,1H),6.22(d,J=8.3Hz,1H),2.33(s,6H),2.23(s,3H).

[0571] Example 94: Synthesis of R554

[0572]

[0573] Step 1: In a 100 mL round-bottom flask, add R554-1 (4.5 g, 22.4 mmol), concentrated sulfuric acid (2.5 g, 24.6 mmol), and methanol (30 mL) in sequence, and heat the reaction system to 50 ° C and stir for 7 hours. Add a saturated solution of sodium carbonate to the reaction solution to neutralize to pH = 8-9, extract with dichloromethane (20 mL x 3), combine the organic phases, dry over anhydrous sodium sulfate, concentrate, and separate the crude product by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound R554-2 (white solid, 3.8 g).

[0574] Step 2: In a 100 mL round-bottom flask, R554-2 (3.5 g, 16.3 mmol), N-chlorosuccinimide (2.4 g, 17.9 mmol), and N,N-dimethylformamide (30 mL) were added in sequence, and the reaction system was heated to 70 ° C and stirred overnight. Water (100 mL) was added to the reaction system, and dichloromethane was extracted (50 mL × 3). The organic phases were combined, washed with water (50 mL x 3) and saturated brine (50 mL) in sequence, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 13:1) to obtain compound R554-3 (white solid, 2.6 g).

[0575] Step 3: In a 50 mL round-bottom flask, compound R554-3 (1.85 g, 7.4 mmol), 3,5-dimethyl-1-(2-tetrahydropyranyl)-1H-pyrazole-4-boronic acid pinacol ester (2.5 g, 8.2 mmol), Pd(dppf)Cl2 (19 mg, 0.02 mmol), potassium carbonate (3.1 g, 22.3 mmol), dioxane / water (30 mL / 5 mL) were added in sequence, heated to 80 ° C and stirred overnight in a nitrogen atmosphere. The reaction solution was concentrated to dryness under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 6:1) to obtain compound R554-4 (yellow solid, 1.5 g), which was directly used in the next step.

[0576] Step 4: In a 50 mL round-bottom flask, R554-4 (350 mg, 1.0 mmol), methyl 2-iodo-5-methylbenzoate (554 mg, 2.0 mmol), Pd2(dba)3 (70 mg, 0.07 mmol), XantPhos (140 mg, 0.2 mmol), Cs2CO3 (490 mg, 1.5 mmol), toluene (25 mL) were added in sequence under nitrogen atmosphere, and the reaction system was heated to 115°C and stirred overnight. The mixture was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound R554-5 (yellow solid, 450 mg).

[0577] Step 5: Compound R554-5 (150 mg, 0.3 mmol) was dissolved in methanol / water (2 mL / 1 mL), lithium hydroxide (145 mg, 6.0 mmol) was added, and the reaction was heated to 50°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound R554-6 (white solid, 58 mg).

[0578] Step 6: In a 25 mL round-bottom flask, compound R554-6 (58 mg, 0.12 mmol), HATU (69 mg, 0.18 mmol) and N,N-dimethylformamide (10 mL) were added in sequence. After stirring at room temperature for 5 minutes, DIEA (47 mg, 0.36 mmol) and ammonium chloride (16 mg, 0.3 mmol) were added. The reaction system was heated to 45 ° C and stirred for 16 hours. Water (20 mL) was added to the reaction system, and dichloromethane was extracted (3×10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure to obtain the crude product R554-7 (yellow solid, 58 mg) which was directly used in the next step.

[0579] Step 7: Dissolve the crude compound R554-7 (58 mg) in dioxane (3 mL), slowly add HCl / dioxane solution (4 M, 1 mL), stir the reaction at room temperature for 24 hours to precipitate solid, filter, rinse with dioxane, and dry to obtain the product compound R554 (hydrochloride, white solid, 40 mg). LCMS: [M+H] + =399; 1 H NMR (600MHz, DMSO-d6) δ7.56–7.54(m,1H),7.52(d,J=2.1Hz,1H),7.41(d,J=2.1Hz,1H),7.05(d d,J=8.4,1.9Hz,1H),6.18(d,J=8.4Hz,1H),4.33(s,2H),3.29(s,3H),2.36(s,6H),2.22(s,3H).

[0580] Example 95: Synthesis of R555

[0581]

[0582] Step 1: In a 50 mL round-bottom flask, R554-4 (226 mg, 0.6 mmol), B-3a (150 mg, 0.5 mmol), Pd2(dba)3 (30 mg, 0.03 mmol), XantPhos (60 mg, 0.1 mmol), Cs2CO3 (243 mg, 0.7 mmol), and toluene (25 mL) were added in sequence under nitrogen atmosphere, and the reaction system was heated to 115°C and stirred overnight. The reaction solution was concentrated to dryness under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound R555-1 (yellow solid, 260 mg).

[0583] Step 2: Compound R555-1 (260 mg, 0.5 mmol) was dissolved in tetrahydrofuran / methanol / water (4 mL / 4 mL / 2 mL), lithium hydroxide (145 mg, 6.0 mmol) was added, and the reaction was heated to 50°C and stirred overnight. The reaction solution was cooled to room temperature, adjusted to pH 5-6 with dilute hydrochloric acid (2 M), and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound R555-2 (white solid, 130 mg).

[0584] Step 3: Compound R555-2 (120 mg, 0.2 mmol) was dissolved in toluene (15 mL), and thionyl chloride (1.5 mL) was slowly added. The reaction system was heated to 100°C and stirred for 4 minutes. The mixture was concentrated to dryness under reduced pressure. The crude product was dissolved in dichloromethane (4 mL), added to ammonia water (4 mL) at 0°C, stirred at room temperature overnight, and concentrated to dryness under reduced pressure. The crude product was separated by column chromatography (dichloromethane: methanol = 20:1) to obtain compound R555 (yellow solid, 20 mg). LCMS: [M+H] + =425; 1 HNMR(600MHz,DMSO-d6)δ12.38(s,1H),9.70(s,1H),8.04(s,1H),7.41–7.31(m,3H),6.98(d,J=8.6Hz,1H), 6.14(d,J=8.5Hz,1H),4.31(s,2H),2.23(s,3H),1.24(s,1H),0.85(d,J=8.3Hz,2H),0.64(d,J=5.4Hz,2H).

[0585] Other compounds were synthesized by referring to the methods described in the above examples.

[0586] Biological Example 1 FTO Inhibitory Activity Assay

[0587] High-purity FTO protein was obtained by nickel column affinity chromatography.

[0588] The FTO enzyme activity inhibition reaction system is as follows: 50 mM Tris·HCl, pH = 7.5, 0.3 μM FTO, 1 μM 39nt-m 6 A modified double-stranded DNA, 300μM 2OG, 280μM (NH4)2Fe(SO4)2, 2mM L-Ascorbic Acid and different concentrations of compounds were incubated at room temperature for 2h, then slowly heated at 65℃ for inactivation, and 1μM 39nt antisense DNA was added to anneal into double strands. 8μL of the reaction solution was taken, and the double-stranded substrate was digested with the methylation-sensitive enzyme DpnII. The digested samples were detected by 15% non-denaturing polyacrylamide electrophoresis, and photographed under the gel imaging system after Gel-Red staining, and the grayscale inhibition rate of the obtained bands was read.

[0589] The following Table 1 shows the inhibition rate of 2-(substituted benzyl) aromatic formic acid and its derivatives represented by general formula (I) at a concentration of 50 μM:

[0590] Table 1 Inhibition rate of FTO enzyme at 50 μM concentration of compounds

[0591]

[0592]

[0593]

[0594] From the test results in Table 1, it can be seen that this type of compound has a strong inhibitory activity on FTO enzyme.

[0595] The following are compounds of 2-(substituted benzoyl) aromatic carboxylic acid and its derivatives represented by general formula (I) with an inhibition rate greater than 70% at a concentration of 50 μM. The enzyme activity inhibition IC values ​​were further determined by PAGE method: 50 The FTO enzyme activity test results of the representative compounds described herein are shown in Table 2.

[0596] Table 2 FTO enzyme activity IC of representative compounds 50

[0597]

[0598]

[0599] From the test results in Table 2, most of these compounds have excellent FTO enzyme inhibitory activity.

[0600] Biological Example 2 Cell Activity Assay

[0601] The following is a study on the cytotoxicity of 2-(substituted benzyl) aromatic formic acid and its derivatives FTO inhibitors represented by general formula (I) on human acute promyelocytic leukemia cells (NB4) and human acute myeloid leukemia cells (Molm13):

[0602] Leukemia cell lines such as NB4 and Molm13 were cultured separately, and cells were seeded at a density of 5000 per well in 96-well plates. The cells were cultured until they adhered to the wall. Different compounds were added and cultured for 72 hours. 10 μL of MTS solution was directly added and incubated for 4 hours. The absorbance at 490 nm was detected, and the inhibition rate was calculated using the DMSO group as the control.

[0603] The following is the FTO inhibitor of 2-(substituted benzyl) aromatic formic acid compounds represented by general formula (I) at 10 μM and 40 μM concentrations. The inhibition rate of NB4 and Molm13 at the 72h time point is greater than 60%. The half inhibition concentration IC is further determined. 50 The anti-cell proliferation activity test results of the representative compounds described herein are shown in Table 3.

[0604] Table 3 Anti-cell proliferation activity of representative compounds

[0605]

[0606]

[0607]

[0608] From the test results in Table 3, most of these compounds have excellent anti-cell proliferation activity.

[0609] Biological Example 3 Cell Activity Assay of Renal Cancer Cell Line 786-O

[0610] The following is a study on the antiproliferative activity of 2-(substituted benzyl) aromatic carboxylic acid and its derivatives as FTO inhibitors represented by general formula (I) against human renal cancer cells (786-O):

[0611] 786-O cells were cultured in 96-well plates at a density of 2500 cells per well. After the cells adhered to the wall, different compounds were added and cultured for 72 hours. After the culture was completed, 10 μL of MTT solution was added and incubated for 4 hours. The absorbance at 490 nm was detected, and the inhibition rate was calculated using the DMSO group as the control.

[0612] The single-point concentrations of the initial screening compounds were 10 μM and 1 μM, and the half-inhibitory concentration (IC 50 The experimental results of the anti-cell proliferation activity of the representative compounds described herein are shown in Table 4:

[0613] Table 4 Anti-cell proliferation activity of representative compounds

[0614] Compound No. <![CDATA[Cell viability (IC 50 )(μM) 786-O]]> R248 0.074 R249 0.34 R250 0.30 R251 0.17 R252 0.22 R253 0.60 R335 0.16 R340 0.76 R308 0.71 R422 0.75 R426 0.74 R427 0.77 R501 0.44 R502 0.69 R506 0.5 R551 0.54 R554 0.70

[0615] Pharmacokinetic evaluation of the compound of biological example 4

[0616] Experimental purpose: Test the pharmacokinetics of compounds in mice

[0617] Experimental materials: ICR mice (male, 4-6 weeks old, Beijing Weitonglihua)

[0618] Experimental procedure: The clear solution obtained after dissolving the test compound was administered to ICR mice (overnight fasting) by oral gavage (2 mg / kg). After oral gavage, about 0.05 mL of blood samples were collected from the cheeks of ICR mice at 0.0833, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours, and plasma was obtained after centrifugation. The drug concentration in mouse plasma samples was determined by LC-MS / MS, and the relevant pharmacokinetic parameters were calculated using WinNolin pharmacokinetic software. The test results are shown in Table 5:

[0619] Table 5 PK test results of compounds in mice

[0620]

[0621]

[0622] Note: 1 / 2 : half-life; C max : peak concentration; T max : Peak time; AUC 0-1ast : Area under the plasma concentration-time curve from time 0 to 24 hours.

[0623] Conclusion: The compounds of the present invention have good oral exposure, and oral administration is conducive to producing good in vivo efficacy.

[0624] Biological Example 5 In vivo pharmacodynamic study of the tail vein transplantation model of human acute myeloid leukemia Molm13

[0625] Experimental purpose: To evaluate the in vivo efficacy of the compounds of the present invention in the tail vein transplantation model of human acute myeloid leukemia Molm13.

[0626] Experimental animals: female NSG mice, 6-8 weeks old; Supplier: Shanghai Jihui Experimental Animal Breeding Co., Ltd.

[0627] Experimental design:

[0628] Table 6 Experimental groups

[0629]

[0630] Note: vehicle: vehicle control group; PO: intragastric administration; QD: once a day;

[0631] Experimental process:

[0632] After irradiation, NSG mice were injected with a certain number of Molm13 cells through the tail vein. After 3 to 7 days, they were randomly divided into groups and given blank solvent (5% DMSO + 5% Tween 80 + 40% PEG400 + 50% saline) or a certain dose of the test article (as shown in Table 6) by gavage once a day. The body weight and survival status of the experimental animals in each group were recorded.

[0633] Experimental results:

[0634] NSG mice were injected with Molm13 cells via the tail vein, and all mice in the control group became ill and died within 2-3 weeks. After treatment with the compounds of the present invention, the onset of the disease in the experimental mice was delayed, and the survival rate and survival time of the mice were significantly prolonged.

[0635] Conclusion: The compounds of the present invention have significant therapeutic effects in the tail vein transplantation model of human acute myeloid leukemia Molm13.

[0636] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound represented by the following formula (I), and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof, in, A1, A2 and A3 are each independently N, CH or CR3; R1 is selected from the following group: C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 3-8 membered heterocyclyl, C3-C8 cycloalkyl-O-, 3-8 membered heterocyclyl-O-; and said R1 may be replaced by one or more R f Substitute, the R f Selected from the group consisting of deuterium, halogen, carbonyl (=O), carboxyl, hydroxyl, amino, nitro, cyano, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkoxycarbonyl, C1-C6 acylamino, C2-C6 ester, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C6~C 10 Aryl, substituted or unsubstituted five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, carbocyclic ring as described in 3-12-membered cycloalkyl; R2 is selected from the following groups: H, halogen atoms, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C 12 Cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 amide, carbonyl, carboxyl, hydroxyl, amino, nitro, cyano; R3 is selected from the following group: H, halogen atoms, hydroxyl, amino, nitro, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 alkoxycarbonyl, substituted or unsubstituted C3-C6 cycloalkyl; X has the following structure: COOH, CONH2, CONHOH, CONHR e 、CONHOR e 、CON(OH)R e 、COOR e ; R e is a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group, or a substituted or unsubstituted 3-8 membered heterocyclic group; R a , R b Each is independently selected from the following group: halogen, -OH, CN, NO2, NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted -(CH2) m -C1~C6 alkoxy, substituted or unsubstituted -(CH2) m -C1~C6 alkylthio, substituted or unsubstituted -(CH2) m -3- to 8-membered heterocyclic group, substituted or unsubstituted -O-(CH2) m -3- to 8-membered heterocyclic group, substituted or unsubstituted C3- to C8-cycloalkyl group, substituted or unsubstituted C3- to C8-cycloalkoxy group, substituted or unsubstituted C6- to C8-cycloalkyl group 10 Aryl, substituted or unsubstituted 5-10 membered heteroaryl; wherein m is selected from the following group: 0, 1, 2, 3, 4, or 5; R c , R d Each is independently selected from the following group: H, halogen, -OH, CN, NO2, NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclic group, substituted or unsubstituted C6-C 10 Aryl, substituted or unsubstituted 5- to 10-membered heteroaryl; Ring A is a group selected from the group consisting of a naphthalene ring, a 3- to 10-membered heterocyclic group, or a 5- to 10-membered heteroaryl group; n is 0, 1, 2, 3 or 4; The substitution mentioned herein refers to the substitution of one or more hydrogen atoms by a substituent selected from the group consisting of deuterium, a halogen atom, a carbonyl group (=O), a carboxyl group, a hydroxyl group, an amino group, a nitro group, a cyano group, a C1-C6 alkoxy group, a deuterated C1-C6 alkoxy group, a halogenated C1-C6 alkoxy group, a C1-C6 alkylamino group, a C1-C6 alkoxycarbonyl group, a C1-C6 acylamino group, a C2-C6 ester group, a C1-C6 alkyl group, a halogenated C1-C6 alkyl group, a C6-C 10 Aryl, five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, C3-C 12 Cycloalkyl; The heterocycle or heterocyclic group may be a saturated or partially unsaturated structure, but not aromatic; the carbocycle, heterocycle, aromatic ring, heteroaromatic ring may be a monocyclic, spirocyclic, fused or bridged ring; the heterocyclic group or heteroaromatic group may each independently contain 1, 2, 3 or 4 heteroatoms selected from O, S and N.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof, characterized in that: R1 is selected from the following group: C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 3-8 membered heterocyclic group, C3-C8 cycloalkyl-O-; and said R1 is optionally replaced by one or more R f Substitute, the R f Selected from the group consisting of deuterium, halogen, carbonyl (=O), carboxyl, hydroxyl, amino, nitro, cyano, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkoxycarbonyl, C1-C6 acylamino, C2-C6 ester, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C6~C 10 Aryl, substituted or unsubstituted five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, carbocyclic ring as described in 3-12-membered cycloalkyl; R a , R b Each is independently selected from the following group: halogen, -OH, CN, NO2, NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted -(CH2) m -C1~C6 alkoxy, substituted or unsubstituted -(CH2) m -C1~C6 alkylthio, substituted or unsubstituted -(CH2) m -3- to 8-membered heterocyclic group, substituted or unsubstituted -O-(CH2) m -3- to 8-membered heterocyclic group, substituted or unsubstituted C3- to C8-cycloalkyl group, substituted or unsubstituted C3- to C8-cycloalkoxy group, substituted or unsubstituted C6- to C8-cycloalkyl group 10 Aryl, substituted or unsubstituted 5-10 membered heteroaryl; wherein m is selected from the following group: 0, 1, or 2; R c , R d Each is independently selected from the following group: H, halogen, C1-C6 alkyl.

3. The compound according to claim 1, and its pharmaceutically acceptable salt, hydrate, solvate or prodrug, characterized in that: The R c and R d Each is independently H.

4. The compound according to claim 1, and its pharmaceutically acceptable salt, hydrate, solvate or prodrug, characterized in that: The X is selected from a substituted or unsubstituted structure selected from the following group: wherein n is 0, 1, 2, 3, 4, 5 or 6; R e R is selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclic group; g Selected from H, OH.

5. The compound according to claim 1, and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof, characterized in that: The A ring is selected from the following group: pyridine, pyrimidine, pyridazine, tetrazine, triazine, pyrrole, thiophene, furan, tetrazole, triazole, imidazole, thiazole, oxazole, pyrazole, isothiazole, isoxazole, oxadiazole, thiadiazole, morpholine, dihydropiperidine, thiomorpholine, piperidine, piperazine, tetrahydropyran, dihydropyran, pyrroline, tetrahydrothiophene, tetrahydrofuran, oxetane, thiazole, and azetidine.

6. The compound according to claim 1, and its pharmaceutically acceptable salt, hydrate, solvate or prodrug, characterized in that: The A ring is selected from the following group:

7. The compound according to claim 1, and its pharmaceutically acceptable salt, hydrate, solvate or prodrug, characterized in that: The R a , R b Each is independently selected from the following group: F, Cl, Br, methyl, ethyl, cyclopropane, isopropyl, methoxy, methylthio, -CH2OCH3, trifluoromethyl, trifluoromethoxy, difluoromethoxy, deuterated methoxy.

8. The compound according to claim 1, and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof, characterized in that: The compound of formula (I) is selected from the following group:

9. A use of a compound of formula (I) as claimed in claim 1 and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof, characterized in that: For use selected from the group consisting of: (a) preparing a drug for treating a disease associated with FTO protein activity or expression; (b) preparing targeted inhibitors of FTO protein activity; (c) non-therapeutic inhibition of FTO protein activity in vitro; and / or (d) Treating diseases related to FTO activity or expression.

10. The use according to claim 9, characterized in that The disease is selected from the following group: leukemia, lymphoma, myelodysplastic syndrome, obesity, metabolic syndrome (MS), type 2 diabetes (T2D), Alzheimer's disease, breast cancer, kidney cancer, colorectal cancer, pancreatic cancer, liver cancer, small cell lung cancer, human bone marrow rhabdomyosarcoma, pancreatic cancer, and malignant glioblastoma.

11. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: (i) an effective amount of a compound of formula (I), and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof; and (ii) a pharmaceutically acceptable carrier.

12. A method for inhibiting FTO protein activity, characterized in that: The method comprises the steps of administering an effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 to the subject of inhibition, or administering an effective amount of the pharmaceutical composition as claimed in claim 11 to the subject of inhibition.

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