A BTK inhibitor

Abstract

The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to a BTK inhibitor; the purpose of the present invention is to analyze the structural characteristics of the marketed drug ibrutinib, and combined with the latest reported structure-activity relationship, the different substituents R1 at the N2 position of piperidine-2,6-dione have little effect on the Btk enzyme activity, and R1 is oriented toward a hydrophilic pocket of the enzyme, and the introduction of different hydrophilic groups can regulate the physicochemical properties of the compound. The spatial orientation of the N9 substituent R2 is oriented toward the gatekeeper region of the kinase, and interacts with specific residues. In the Tec family, there are differences in the amino acid residues that interact with the gatekeeper region of different kinases: it is expected to design and synthesize more cytotoxic compounds, exert anti-cancer effects, and have drug-forming potential. Cytotoxicity experiments have found that the compounds of the present invention have a strong in vitro inhibitory effect on a variety of cancer cells.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to a novel 3-amino-2,7-dihydropyrano[4,3-c]pyrazole-4,6-dione BTK inhibitor, and a preparation method and use of the inhibitor. Background Art

[0002] Tumor is a common and frequently-occurring disease that poses a serious threat to human health. It is caused by the abnormal proliferation of cells in the human body. Lung cancer, digestive tract cancer and liver cancer are the most common tumors in men, accounting for more than 70% of all cases (lung cancer 23%, stomach cancer 15.2%, liver cancer 13.57%, esophageal cancer 10.46%, colorectal cancer 9.39%), while breast cancer, lung cancer, digestive tract cancer and liver cancer are the most common tumors in women, accounting for more than 60% of all cases (breast cancer 16.97%, lung cancer 14.85%, colorectal cancer 9.68%, stomach cancer 8.53%, liver cancer 6.17%). With the aging of China's population, the prevalence of cancer is bound to increase. In addition, environmental pollution has also increased the prevalence of some cancers, such as lymphoma. Autoimmune diseases are caused by the immune response of the immune system to the components of the body itself, causing damage and leading to disease.

[0003] BTK (brutontyrosine kinase) is a cytoplasmic protein belonging to the non-receptor tyrosine kinase Tec family. It is expressed in most hematopoietic cells, such as B cells, mast cells, megakaryocytes, etc., but not in T cells, NK cells and plasma cells. BTK plays a very critical role in the B cell receptor signaling pathway and has an important impact on the proliferation, differentiation and apoptosis of B cells.

[0004] In malignant B cells, the B cell receptor signaling pathway is overactive, thereby inhibiting the normal differentiation and apoptosis of B cells and promoting abnormal proliferation. It is known that abnormal regulation of the BCR pathway often exists in malignant tumors of various B cell types, such as diffuse large B cell lymphoma (DLBCL), mantle cell lymphoma (MCL), chronic lymphocytic leukemia (CLL), follicular lymphoma (FL), Waldenstrom's macroglobulinemia (WM), marginal zone lymphoma (MZL), B-lymphoblastic leukemia (B-LBL) and Burkitt's lymphoma (BL).

[0005] Since the BTK inhibitor was first discovered in 1993, drug research on BTK has never stopped. The indications of BTK inhibitors are still expanding. In addition to anti-tumor, the application of autoimmune diseases will be the key to pushing BTK inhibitors to another market height. At present, a variety of small molecule inhibitors of BTK have been developed in China and have shown good anti-tumor effects in in vivo and in vitro tests. In addition to the marketed drugs Ibrutinib and ACP-196, more new BTK inhibitors are under research. In addition to BGB-3111, CT1530 and GS-4059 have entered Phase II clinical trials.

[0006] At present, the second generation of tumor-targeted small molecule drugs targeting BTK has been developed. The first generation of BTK inhibitors is Ibrutinib, which was launched in the United States in 2013 for the clinical treatment of relapsed or refractory mantle cell lymphoma (MCL). It is a star drug targeting BTK. Ibrutinib can inhibit the growth and metastasis of malignant proliferating B cells. At the same time, Ibrutinib is the first BTK inhibitor approved by the FDA. In addition to being used to treat various blood diseases, Ibrutinib is also used to treat autoimmune-related diseases including stem cell transplantation, post-transplant immune resistance and arthritis. However, the response rate of Ibrutinib in treatment is not 100%, and the disease will relapse. The drug has unknown primary resistance and acquired resistance. With the listing and clinical use of the drug, Ibrutinib began to produce different mechanisms of resistance in different patients. Acquired drug resistance has only been found in these two target mutations so far. In the BCR signaling pathway, a mutation occurred at position 481 of the binding site between BTK and ibrutinib. Ibrutinib dose-dependently inhibits the disease in the arthritis model. There are more and more problems of drug resistance to Ibrutinib in clinical practice, and many side effects have also occurred.

[0007] Acalabrutinib (ACP-196), the second-generation BTK inhibitor, is called the second-generation BTK inhibitor because it does not inhibit other kinases on the basis of inhibiting the BTK kinase target. It was approved by the FDA in August 2017 for the treatment of mantle cell lymphoma (MCL). There are many indications in clinical trials, such as chronic lymphocytic leukemia (CLL) in Phase III clinical trials; some solid tumors such as non-small cell lung cancer and head and neck cancer are in Phase II clinical trials, and the development is very rapid. ACP-196 is also currently in clinical trials for the treatment of arthritis because of the off-target effect of Ibrutinib. Therefore, the second-generation BTK inhibitor with better selectivity has begun to emerge. Due to its high selectivity and few side effects, it has good development prospects in the treatment of chronic diseases such as autoimmune diseases.

[0008]

[0009] It is known that the selectivity of BTK inhibitors is not ideal. In addition to inhibiting BTK, they also inhibit a variety of other kinases (such as ETK, EGF, BLK, FGR, HCK, YES, BRK and JAK3, etc.), resulting in more side effects. At the same time, mutations in BTK binding sites often lead to drug resistance. Therefore, more BTK inhibitors are needed clinically to treat diseases such as tumors and overcome such adverse events.

[0010] The present invention designs and synthesizes a novel 3-amino-2,7-dihydropyrano[4,3-c]pyrazole-4,6-dione compound. In vitro experiments show that the compound has good cytotoxicity to cancer cells, among which compound TM10 has an IC of 1.0447 / cm3 on lung cancer cells. 50 The concentration of the compound reaches 0.86 μM, and the compound is a potential compound for development as a new anticancer drug. In terms of preparation method, the present invention efficiently synthesizes key intermediates and target compounds through a simple method. Summary of the invention

[0011] The purpose of this invention is to analyze the structural characteristics of the marketed drug ibrutinib, combined with the latest reported structure-activity relationship, the different substituents R1 at the N2 position of piperidine-2,6-dione have little effect on the activity of Btk enzyme, and R1 faces a hydrophilic pocket of the enzyme. The introduction of different hydrophilic groups (such as acid, amide, amine) can adjust the physicochemical properties of the compound. The spatial orientation of the substituent R2 at the N9 position faces the gatekeeper region of the kinase and interacts with specific residues. In the Tec family, the amino acid residues that interact with the gatekeeper region of different kinases are different:

[0012] It is expected that the compound with higher cytotoxicity can be designed and synthesized to exert anti-cancer effect and have drug-forming potential. Cytotoxicity experiments have found that the compound of the present invention has a strong in vitro inhibitory effect on a variety of cancer cells.

[0013] Explanation of terms

[0014] Listed below are definitions of various terms used to describe this application. These definitions apply to the terms as used throughout the specification and claims, unless otherwise limited in specific cases, either individually or as part of a larger group.

[0015] The term "alkyl" in the present invention refers to a saturated straight or branched hydrocarbon group, which, in certain embodiments, contains 1 to 4 carbon atoms, 1-4 Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, or the like.

[0016] The term "aryl" in the present invention refers to a monocyclic or polycyclic carbon ring system having one or more fused or non-fused aromatic rings, including but not limited to phenyl, naphthyl, tetrahydronaphthyl and the like, and the hydrogen atoms on the ring carbons may be substituted by one or more substituents.

[0017] The term "halogenated" in the present invention refers to a group formed by a halogen atom replacing a hydrogen atom on a carbon atom, wherein the halogen atom includes but is not limited to F, Cl, Br, and I.

[0018] The term "alkoxy" in the present invention refers to -O-alkyl, wherein the alkyl is not limited to C 1-3 Alkyl and C 3-6 Specific examples of cycloalkyl include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and halogenated forms thereof.

[0019] The term "alkylamino" in the present invention refers to the group -N-alkyl, wherein the alkyl group is not limited to C 1-3 Alkyl and C 3-6 Cycloalkyl, specific examples include but are not limited to methylamino, ethylamino, propylamino, cyclopropylamino, cyclobutyloxy and the like. The term "cycloalkyl" of the present invention refers to a monovalent group of a monocyclic or polycyclic saturated or partially unsaturated carbocyclic compound, C 3-6 Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like, and the hydrogen atoms on the ring carbons may be substituted by one or more substituents.

[0020] The term "heterocycloalkyl" of the present invention refers to a monocyclic or polycyclic non-aromatic ring system containing 2 to 6 ring carbon atoms and 1 to 3 ring heteroatoms, wherein the heteroatoms are selected from N, O, and S.

[0021] The term "heteroaryl" of the present invention refers to an aromatic ring system containing 1 to 6 carbon atoms and at least one heteroatom, wherein the heteroatom is selected from N, S, and O.

[0022] The term "alkylamino" herein refers to the group -alkyl-NR a R b , wherein the alkyl group includes but is not limited to C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy and C 3-6 Cycloalkyl, R a and R b Each independently selected from H, C 1-3 Alkyl or C 3-6 Cycloalkyl.

[0023] The "pharmaceutically acceptable salts" of the present invention refer to salts of the compounds of the present invention, which are prepared by reacting the compounds with specific substituents discovered by the present invention with relatively non-toxic acids or bases. The compounds of the present invention contain relatively basic functional groups, and acid addition salts can be obtained by contacting the neutral form of such compounds with sufficient acid in a pure solution or a suitable inert solvent. Pharmaceutically acceptable acid addition salts include inorganic salts and organic acid salts. The inorganic acid salts include, but are not limited to, hydrochlorides, nitrates, borates, hydrocyanates, hydrofluorides, hydrobromides, hydroiodides, nitrites, perhalates, halides, halites, hypohalites, aluminates, sulfates, phosphates, and nitrates; the organic acid salts include, but are not limited to, formates, acetates, propionates, butyrates, acrylates, oxalates, malonates, succinates, benzoates, phthalic acid carboxylates, methanesulfonates, ethanesulfonates, benzenesulfonates, toluenesulfinates, thioacetates, trifluoroacetates, tartrates, malates, citrates, ascorbates, salicylates, caffeic acid fumarates, maleates, lactates, citrates, glutamates, camphorates, camphorsulfonates, and the like.

[0024] The "solvate" described in the present invention refers to a solvent addition form containing stoichiometric or non-stoichiometric amounts, and the solvent can be selected from water, ethanol, isopropanol, ether, acetone, and the like.

[0025] The "prodrug" of the present invention refers to a compound that can be converted in vivo by metabolism to provide any compound described by the chemical formula of the present application, and various forms of drugs are known in the art.

[0026] The "effective amount" mentioned in the present invention refers to a dose that can achieve the desired therapeutic effect in the desired subject without causing excessive negative effects. The specific dose can usually be determined by technicians in this field according to needs.

[0027] The term "treatment" as used herein refers to a method of alleviating or alleviating the symptoms of a disease and its complications; the term "prevention" as used herein refers to reducing or eliminating the symptoms or onset of complications of a disease, condition or disorder.

[0028] It should be understood that other terms not explained above but appearing in the present invention should be defined according to the common understanding of those skilled in the art.

[0029] The specific technical solutions of the present invention are as follows:

[0030] In a first aspect, the present invention provides a hydrate, solvate, prodrug, stereoisomer or tautomer of a pyrazole-4,6-dione compound or a pharmaceutically acceptable salt thereof represented by formula (I):

[0031]

[0032]

[0033] in:

[0034] R is independently selected from H, halogen, aryl, heteroaryl, C 1-4 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 2-6 heterocycloalkyl, aryloxy, -NR2R3; or substituted aryl, substituted heteroaryl, substituted C 3-6 Cycloalkyl, substituted C 2-6 Heterocycloalkyl, substituted aryloxy, substituted C 1-6 One or more independent groups in an alkoxy group, wherein the substituent group is selected from halogen, aryl, heteroaryl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 1-4 Alkyl, substituted C 2-6 Heterocycloalkyl, halogen-substituted C 2-6 Heterocycloalkyl, C 2-6 One or more substituents in the heterocycloalkyl group.

[0035] R is preferably -NR2R3, aryl, heteroaryl, C 1-4 Alkyl, C 2-6 Heterocycloalkyl, substituted aryl, substituted heteroaryl.

[0036] More preferably, R is -NR2R3, an aryl group, a heteroaryl group, or a substituted aryl group.

[0037] R2 is independently selected from H or C 1-4 Alkyl; R2 is more preferably methyl.

[0038] R3 is independently selected from H, C 1-4Alkyl, C 3-6 Cycloalkyl, C 1-3 Haloalkyl, aryl, heteroaryl, C 1-6 Alkoxy, C 1-6 Alkylamino, substituted C 2-6 Heterocycloalkyl, halogen-substituted C 2-6 Heterocycloalkyl, C 2-6 One or more of the heterocycloalkyl groups; R3 is further preferably methyl.

[0039] Z is independently selected from C 1-4 An alkyl group or an aryl group; Z is preferably a methyl group, an ethyl group, or a benzene group; and Z is more preferably a methyl group or a benzene group.

[0040] In a preferred embodiment of the present invention, the compound of formula (I) is selected from the following group:

[0041]

[0042]

[0043] In a second aspect of the present invention, a method for preparing a pyrazole-4,6-dione compound of formula (I) is provided. The compounds of the present application can be prepared by using a variety of methods known to those skilled in the art. The present application can be synthesized using the methods described herein and organic chemical synthesis methods or variations thereof understood by those skilled in the art. Preferred methods include but are not limited to the following methods.

[0044] Preferably, the method for preparing the pyrazole-4,6-dione compound represented by formula (I) of the present invention comprises the following steps:

[0045] Preparation of compound 6:

[0046]

[0047] General synthesis method of compound 11:

[0048]

[0049] Preparation of Compound I:

[0050]

[0051] Wherein, R and Z are defined as the compound of formula (I) of the first aspect of the present invention;

[0052] Preferably, the method for preparing the pyrazole-4,6-dione compound represented by formula (I) comprises the following reaction steps:

[0053] Step 1: Compound 1 undergoes condensation reaction in sodium ethoxide solution to obtain compound 2;

[0054] Step 2: Compound 2 reacts with hydrazine hydrate to obtain compound 3;

[0055] Step 3: Compound 3 reacts under alkaline conditions to obtain compound 4;

[0056] Step 4: Compound 4 is subjected to temperature-controlled reflux dehydration to generate intermediate compound 5;

[0057] Step 5: Compound 5 is reacted with 3-bromo-4-fluorobenzoic acid, EDCI and HOBt in dichloromethane to obtain compound 6.

[0058] Step 6: Compound 8 and compound 9 react to obtain compound 10.

[0059] Step 7: Compound 10 is hydrolyzed to obtain compound 11.

[0060] Step 8: Compound 6 and compound 11 react in an organic solvent, crystallize, filter and dry to obtain the target compound I. Preferably, the preparation method of the pyrazole-4,6-dione compound represented by formula (I) comprises the following specific reaction steps:

[0061] Step 1: Add sodium ethoxide to ethanol solvent, stir to dissolve, add compound 1, raise the temperature to reflux, detect by TLC, and after the reaction is completed, control the temperature for crystallization, filter, rinse the filter cake with ethanol, and dry to obtain a crude compound 2; the crude product is recrystallized to obtain the final compound 2.

[0062] Preferably, the crystallization temperature is 15-30°C.

[0063] Step 2: Add compound 2 to water, stir to dissolve, add hydrazine hydrate, raise the temperature to reflux, detect by TLC, when the reaction is complete, cool down to precipitate solid, filter and dry to obtain compound 3.

[0064] Preferably, the crystallization temperature is -10 to 5°C.

[0065] Step 3: Compound 3 is added to the alkaline solution, stirred to dissolve, heated to reflux, and detected by TLC. When the reaction is complete, the temperature is lowered and hydrochloric acid is added to adjust the pH to 3-4. The solid is precipitated, filtered, and dried to obtain compound 4.

[0066] Preferably, the base is sodium hydride, potassium hydroxide, sodium hydroxide, lithium hydroxide, more preferably sodium hydroxide.

[0067] Step 4: Dissolve compound 4 in acetic anhydride, add a condensing agent, heat to reflux, and monitor by TLC. The reaction is complete.

[0068] The mixture was cooled and crystallized, filtered, the filter cake was rinsed with methyl tert-butyl ether, and dried to obtain compound 5.

[0069] Preferably, the condensing agent is dicyclohexylcarbodiimide (DCC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride [EDCI], 2-chloro-1,3-dimethylimidazoline chloride (DMC), and more preferably dicyclohexylcarbodiimide (DCC).

[0070] Step 5: Dissolve 3-bromo-4-fluorobenzoic acid in dried dichloromethane, add EDCI, HOBt and compound 5 in sequence, stir to react at room temperature, detect by TLC, and after the reaction is completed, filter, concentrate, evaporate to dryness under reduced pressure, and separate by column chromatography to obtain compound 6.

[0071] Step 6: Compound 8 is added to dried dichloromethane, and compound 9 and a condensing agent are added in sequence, stirred at room temperature, and tested by TLC. After the reaction is completed, the mixture is evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10.

[0072] Preferably, the condensing agent is selected from dicyclohexylcarbodiimide (DCC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride [EDCI], 2-chloro-1,3-dimethylimidazoline chloride (DMC), 1-carbonylbenzotriazole (HOBt), and more preferably 1-carbonylbenzotriazole (HOBt) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride [EDCI].

[0073] Step 7: Dissolve compound 10 in an organic solvent, add 3 mol / L hydrochloric acid dropwise to remove the Boc group, wash with purified water, dry the organic layer over anhydrous sodium sulfate, filter, concentrate, and evaporate to dryness under reduced pressure to obtain compound 11.

[0074] Preferably, the organic solvent is ethyl acetate, dichloromethane, methanol, ethanol, among which ethyl acetate is preferred.

[0075] Step 8: Compound 6 is dissolved in an organic solvent, compound 11 is added, and the mixture is heated to reflux for reaction. The reaction is completed by TLC detection. The mixture is cooled for crystallization, the filter cake is washed with an organic solvent, and dried to obtain compound I.

[0076] Preferably, the organic solvent is toluene, benzene, or xylene, with toluene being preferred.

[0077] Preferably, the crystallization temperature is -5 to 5°C, preferably 0°C.

[0078] The third aspect of the present invention provides a pharmaceutical composition, which comprises: a therapeutically effective amount of a compound of formula (I) according to the present invention or one or more of its pharmaceutically acceptable salts, solvates, prodrugs, stereoisomers or tautomers, and optionally a pharmaceutically acceptable carrier, excipient, adjuvant, auxiliary material or diluent.

[0079] In a preferred embodiment, the pharmaceutical composition can be used to treat diseases associated with abnormal expression of tyrosine kinase or high activity of tyrosine kinase.

[0080] In a preferred embodiment, the diseases associated with abnormal tyrosine kinase expression or high tyrosine kinase activity include but are not limited to abnormal cell proliferation, morphological changes, hypermotility, angiogenesis, tumor growth, and tumor metastasis.

[0081] A preferred embodiment provides a BTK inhibitor, which contains an inhibitory effective amount of a pyrazole-4,6-dione compound of formula (I) as described in the present invention, or one or more of its pharmaceutically acceptable salts, tautomers, optical isomers, pharmaceutically acceptable solvates, and optionally a pharmaceutically acceptable carrier, excipient, adjuvant, auxiliary material or diluent.

[0082] The compounds of the present invention can be used directly for prevention and treatment, or preferably in the form of a pharmaceutical composition. Although the active ingredient can be administered alone, it is preferably in the form of a pharmaceutical preparation or composition. Therefore, the present invention provides a pharmaceutical preparation comprising a compound of the present invention and a pharmaceutically acceptable diluent, excipient or carrier (collectively referred to herein as "carrier" material). The pharmaceutical composition of the present invention can take the form of a pharmaceutical preparation described below. Therefore, the present invention relates to a pharmaceutical composition comprising at least one compound of formula (I) and a conventional excipient.

[0083] Exemplary compositions for oral administration include: suspensions, which may include, for example, microcrystalline cellulose for imparting volume, alginic acid or sodium alginate as suspending agents, methylcellulose as viscosity enhancers, and sweeteners or flavorings, such as those known in the art; such as immediate release tablets, which may include, for example, microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate, calcium sulfate, sorbitol, glucose and / or lactose and / or other excipients, binders, expanders, disintegrants, diluents and lubricants, such as those known in the art. Suitable binders include: starch, gelatin, natural sugars such as glucose or β-lactose, corn sweeteners, natural and synthetic gums such as gum arabic, tragacanth or sodium alginate, carboxymethyl cellulose, polyethylene glycol and wax, etc. Disintegrants include, but are not limited to: starch, methylcellulose, agar, bentonite and xanthan gum, etc. The compound of formula (I) can also be delivered orally by sublingual and / or buccal administration. Molded tablets, compressed tablets or lyophilized tablets are exemplary forms that can be used. Exemplary compositions include those in which the compounds of the invention are formulated with fast-dissolving diluents such as mannitol, lactose, sucrose and / or cyclodextrin. These preparations may also include high molecular weight excipients such as cellulose (microcrystalline powdered cellulose) or polyethylene glycol (PEG). These preparations may also include excipients that contribute to mucosal adhesion, such as hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), sodium carboxymethyl cellulose (SCMC), maleic anhydride copolymers (e.g., Gantrez) and controlled release agents such as polyacrylic acid copolymers (e.g., Carbopol 934). Lubricants, glidants, flavoring agents, colorants and stabilizers may also be added for ease of preparation and use. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate and sodium chloride, etc. For oral administration in liquid form, the oral drug component may be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol and water.

[0084] Pharmaceutical formulations of the present invention include those suitable for oral, parenteral [including subcutaneous, intradermal, intramuscular, intravenous (bolus or infusion) and intraarticular], inhalation (including fine particle powders or sprays that can be produced by means of various types of metered pressurized aerosols), nebulizer or inhaler, rectal, intraperitoneal and topical (including dermal, buccal, sublingual and intraocular) administration, although the most appropriate route may depend, for example, on the condition and state of the recipient.

[0085] The formulations of the present invention suitable for oral administration may be in the form of various individual units containing a predetermined amount of the active ingredient, such as capsules, cachets, pills or tablets; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids, such as elixirs, tinctures, suspensions or syrups; or oil-in-water emulsions or water-in-oil emulsions. The active ingredient may also be formulated as a bolus, electuary or paste.

[0086] Tablets can be made by compression or molding with one or more optional auxiliary ingredients. The method of making compressed tablets can be to compress the active ingredients in free-flowing forms such as powders or granules, which are optionally mixed with binders, lubricants, inert diluents, lubricants, surfactants or dispersants in a suitable machine. Molded tablets can be molded by a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine. The tablets can be optionally coated or scored and can be formulated to provide a sustained or controlled release of the active ingredient therein. The compounds of the present invention can be administered in a form suitable for immediate release or sustained release. Immediate release or sustained release can be achieved by using a suitable pharmaceutical composition containing the compounds of the present invention, or, especially in the case of sustained release, by using a device such as a subcutaneous implant or an osmotic pump. The compounds of the present invention can also be administered in a liposome manner. A preferred unit dose formulation is a formulation of an active ingredient containing an effective dose (as described below) or a suitable portion thereof.

[0087] It should be understood that in addition to the ingredients specifically mentioned above, the formulations of the present invention may also include other conventional agents in the art related to the type of formulation in question, for example, formulations suitable for oral administration may include flavoring agents.

[0088] The preparations can be conveniently provided as unit dosage forms and can be manufactured by any method well known in the pharmaceutical field. All methods include the step of combining the active ingredient with a carrier constituting one or more additional ingredients. Typically, the active ingredient is uniformly and intimately combined with a liquid carrier and / or a subdivided solid carrier to prepare the preparation, and then, if necessary, the product is molded to obtain the desired preparation.

[0089] The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from various phospholipids, 1,2-dipalmitoylphosphatidylcholine, phosphatidylethanolamine (cephalin), phosphatidylserine, phosphatidylinositol, diphosphatidylglycerol (cardiolipin) or phosphatidylcholine (lecithin).

[0090] Preparations for parenteral administration include aqueous and non-aqueous sterile injection solutions, which may contain antioxidants, buffers, antibacterial agents, and solutes that make the preparation isotonic with the blood of the designated recipient; and aqueous and non-aqueous sterile suspensions, which may contain suspending agents and thickening agents. The preparation can be provided in unit dose or multi-dose containers, such as sealed ampoules and vials, and the composition can also be preserved by freeze drying (lyophilization) conditions, and only a sterile liquid carrier such as saline or water for injection needs to be added before use. Extemporaneous injection solutions and suspensions can be prepared from the sterile powders, granules, and tablets described above. Exemplary compositions for parenteral administration include: injectable solutions or suspensions, which may contain, for example, suitable non-toxic parenteral acceptable diluents or solvents, such as polyethylene glycol, ethanol, 1,3-butanediol, water, Ringer's solution, sodium chloride isotonic solution, or other suitable dispersants or wetting agents and suspending agents, including synthetic mono- or diglycerides and fatty acids, including oleic acid and Cremaphor.

[0091] Exemplary compositions for nasal, aerosol, or inhaled administration include solutions in saline, which may contain, for example, benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, and / or other solubilizing or dispersing agents such as those known in the art.

[0092] Formulations for rectal administration can be made into suppositories using conventional carriers such as cocoa butter / synthetic glycerides or polyethylene glycol. These carriers are usually solid at room temperature but liquefy and / or dissolve in the rectal lumen to release the drug.

[0093] Formulations for topical administration in the mouth, e.g., buccal or sublingual administration, include lozenges comprising the active ingredient in a flavored base such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a base of gelatin and glycerin or sucrose and acacia. Exemplary compositions for topical administration include a topical carrier such as Plastibase (mineral oil gelled with polyethylene).

[0094] In a fourth aspect, the present invention provides a use of a pyrazole-4,6-dione compound of formula (I), specifically, it can be used to prepare a tyrosine kinase inhibitor, for in vitro non-therapeutic inhibition of tyrosine kinase activity, for in vitro non-therapeutic inhibition of tumor cell growth or a combination thereof, and its use in the treatment or prevention of diseases in which BTK plays a role.

[0095] In a preferred embodiment, the pyrazole-4,6-dione compound of formula (I) of the present invention is used for preparing a drug for treating or preventing a disease in which BTK plays a role.

[0096] In a preferred embodiment, the diseases in which BTK plays a role include cancer or autoimmune system-related diseases, and the specific cancers include but are not limited to non-small cell lung cancer, lymphoma, breast cancer, leukemia and head and neck squamous cell carcinoma sarcoma, lung cancer, ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, plasmacytoma, extranodal marginal zone B cell lymphoma, lymph node marginal zone B cell lymphoma, non-Hodgkin lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, renal cancer, anaplastic large cell lymphoma , acute myeloid leukemia, multiple myeloma, melanoma or mesothelioma; the autoimmune-related diseases include but are not limited to chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, Goodpasture's syndrome, primary biliary cirrhosis, multiple sclerosis, acute idiopathic polyneuritis, systemic lupus erythematosus, rheumatoid arthritis, systemic vasculitis, scleroderma, herpes simplex, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia, thyroid autoimmune disease, and ulcerative colitis.

[0097] Preferably, the treatment refers to a method of alleviating or relieving the symptoms of a disease and its complications; the prevention refers to reducing or eliminating the symptoms or onset of complications of a disease, condition or disorder.

[0098] Compared with the prior art, the main advantages of the present invention include:

[0099] (1) The present invention provides a novel pyrazole-4,6-dione compound of formula (I).

[0100] (2) The pyrazole-4,6-dione compounds of formula (I) provided by the present invention have a significant inhibitory effect on the growth of tumor cells.

[0101] (3) The pyrazole-4,6-dione compound of formula (I) provided by the present invention has cheap and readily available starting materials, a short synthetic route, simpler operation, and is suitable for industrial scale-up production.

[0102] 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 (including embodiments) can be combined with each other to form new or preferred technical solutions. DETAILED DESCRIPTION

[0103] The method of the present invention is further illustrated by the following examples. It should be correctly understood that the preparation methods of the examples of the present invention are only used to illustrate the present invention, rather than to limit the present invention, and simple improvements to the preparation methods of the present invention under the premise of the concept of the present invention belong to the scope of protection claimed in the present invention.

[0104] Abbreviations used in this article:

[0105] DCC: dicyclohexylcarbodiimide

[0106] EDCI: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride

[0107] DMC: 2-chloro-1,3-dimethylimidazoline chloride

[0108] HOBt: 1-Carbonylbenzotriazole

[0109] TLC: Thin layer chromatography

[0110] Boc group: tert-butyloxycarbonyl

[0111] Preparation of compound 6:

[0112]

[0113] Preparation of compound 2: Add sodium ethoxide solid (4 g, 0.01 mol) to anhydrous ethanol (40 ml), stir at room temperature to dissolve, pour in (13.20 g, 0.02 mol) of malononitrile, slowly raise the temperature to reflux, detect by TLC, and after the reaction is completed, control the temperature to 20°C, stir and crystallize, after crystallization is completed, filter, rinse with ethanol, and transfer the obtained solid to a three-necked flask after drying under reduced pressure. Pour in purified water, stir at room temperature until dissolved, adjust the pH to 3-4 with 5 mol / L hydrochloric acid, precipitate crystals, filter, and recrystallize the filter cake with purified water to obtain white needle-shaped crystals, which are dried under reduced pressure to obtain compound 2 with a yield of 85.6% and a purity of 99.92%.

[0114] Preparation of compound 3:

[0115] Add intermediate 2 to 500 ml of water, stir to dissolve, add 50 ml of hydrazine hydrate, slowly raise the temperature to reflux, detect by TLC, the reaction is complete, cool to 0°C for crystallization, filter the precipitate, and dry under reduced pressure to obtain compound 3 with a yield of 87.8% and a purity of 99.89%.

[0116] Preparation of compound 4:

[0117] Compound 3 was added to 0.2M NaOH (200 ml), stirred to dissolve, heated to reflux for reaction, and detected by TLC. After the reaction was completed, the temperature was lowered to 0°C, and the reaction solution was adjusted to pH 3-4 with 5 mol / L hydrochloric acid, filtered, and dried under reduced pressure to obtain intermediate 4 with a yield of 92.5% and a purity of 99.90%.

[0118] Preparation of compound 5:

[0119] Compound 4 (150 g, 0.90 mol) was dissolved in acetic anhydride (500 ml), and DCC (240.5 g, 1.17 mol) was added. The mixture was heated to reflux and tested by TLC. After the reaction was completed, the mixture was cooled to 0°C for crystallization and filtered. The filter cake was washed with methyl tert-butyl ether and dried under reduced pressure to obtain compound 5 with a yield of 91.7% and a purity of 99.88%.

[0120] Preparation of compound 6:

[0121] 3-Bromo-4-fluorobenzoic acid (2.19 g, 0.1 mol) was added to 300 ml of dichloromethane, and EDCI (1.92 g, 0.1 mol), HOBt (1.35 g, 0.1 mol) and intermediate 5 (83.6 g, 0.08 mol) were added in sequence. The reaction was stirred at room temperature and detected by TLC. After the reaction was completed, the mixture was filtered, concentrated, and evaporated to dryness under reduced pressure. Compound 6 was obtained by column chromatography with a yield of 90.5% and a purity of 99.87%.

[0122] Example 1

[0123] Preparation of compound TM1:

[0124]

[0125] Preparation of compound 10-1:

[0126] Compound 8 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-1 (8.90 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-1 with a yield of 91.8% and a purity of 99.78%.

[0127] Preparation of compound 11-1:

[0128] Compound 10-1 (23.13 g, 0.1 mol) was dissolved in 150 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The organic layer was washed with purified water and dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11 with a yield of 90.3% and a purity of 99.85%.

[0129] Preparation of compound TM1:

[0130] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-1 (15.74 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM1 with a yield of 92.4% and a purity of 99.92%.

[0131] 1 HNMR (400MHz, DMSOd6): δ12.15(s,1H),9.76(s,1H),8.98(s,1H),7.82~7.74(m,3H),4.66(s,2H),4.32(s,2H),3.42(s,2H),2.76(s,6H);

[0132] 13 CNMR(100MHz,DMSOd6):173.4,168.5,164.6,159.8,155.5,150.2,141.1,133.4,133.5,128.9,118.7,112.2,98.7,78.1,54.8,45.5(2C),30.4.

[0133] MS (m / z): 481.24 [M+H] + ;

[0134] Example 2

[0135] Preparation of compound TM2:

[0136]

[0137] Preparation of compound 10-2:

[0138] Compound 8 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-2 (12.89 g, 0.13 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-2 with a yield of 91.5% and a purity of 99.77%.

[0139] Preparation of compound 11-2:

[0140] Compound 10-2 (25.64 g, 0.1 mol) was dissolved in 150 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The organic layer was washed with purified water and dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-2 with a yield of 91.1% and a purity of 99.89%.

[0141] Preparation of compound TM2:

[0142] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-2 (18.75 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM2 with a yield of 92.1% and a purity of 99.90%.

[0143] 1 HNMR(400MHz,DMSO-d6)δ12.15(s,1H),9.76(s,1H),8.98(s,1H),7.82~7.74(m,3H),4.35(s, 2H),3.92(s,2H),3.55~3.47(m,1H),1.74~1.69(m,2H),1.50~1.43(m,4H),1.23~1.11(m,4H);

[0144] 13 C-NMR(100MHz,DMSO-d6):168.4,157.6,155.5,150.2,141.1,133.4,132.6,129. 5,120.9,118.7,112.2,98.7,54.8,50.1,35.6(2C),30.5,27.0,25.6,26.7(2C).

[0145] MS (m / z): 506.12 [M+H] + ;

[0146] Example 3

[0147] Preparation of compound TM3:

[0148]

[0149] Preparation of compound 10-3:

[0150] Compound 8 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-3 (11.18 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-3 with a yield of 92.0% and a purity of 99.82%.

[0151] Preparation of compound 11-3:

[0152] Compound 10-3 (25.03 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-3 with a yield of 91.6% and a purity of 99.90%.

[0153] Preparation of compound TM3:

[0154] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-3 (18.02 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM3 with a yield of 92.1% and a purity of 99.90%.

[0155] 1 HNMR (400MHz, DMSO-d6) δ12.15(s,1H),9.76(s,1H),8.98(s,1H),7.82~7.74(m,3H),7.23~7.03(m,5H),4.65(s,2H),3.42(s,2H);

[0156] 13 C-NMR(100MHz,DMSO-d6):168.4,157.6,155.5,150.2,141.1,137.1,133.4,134.3, 130.4,129.5,128.8(2C),127.5,121.8(2C),120.9,118.7,112.2,98.7,54.8,30.5.

[0157] MS (m / z): 500.18 [M+H] + ;

[0158] Example 4

[0159] Preparation of compound TM4:

[0160]

[0161] Preparation of compound 10-4:

[0162] Compound 8 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-4 (13.33 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-4 with a yield of 92.2% and a purity of 99.85%.

[0163] Preparation of compound 11-4:

[0164] Compound 10-4 (26.83 g, 0.1 mol) was dissolved in 150 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The organic layer was washed with purified water and dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-4 with a yield of 91.8% and a purity of 99.91%.

[0165] Preparation of compound TM4:

[0166] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-4 (20.18 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM4 with a yield of 91.7% and a purity of 99.87%.

[0167] 1 HNMR (400MHz, DMSO-d6) δ12.15(s,1H),9.76(s,1H),8.98(s,1H),7.82~7.74(m,3H),7.23~7.03(m,4H),4.65(s,2H),3.42(s,2H);

[0168] 13 C-NMR(100MHz,DMSO-d6):168.4,162.9,157.6,155.5,150.2,141.1,137.1,133.4, 134.3,130.4,129.5,128.8(2C),127.5,121.8(2C),118.7,112.2,98.7,54.8,30.5.

[0169] MS (m / z): 518.31 [M+H]+ ;

[0170] Example 5

[0171] Preparation of compound TM5:

[0172]

[0173] Preparation of compound 10-5:

[0174] Compound 8 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-5 (11.29 g, 0.12 mol), HOBt (13.51 g, 0.1 mol) and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-5 with a yield of 91.9% and a purity of 99.81%.

[0175] Preparation of compound 11-5:

[0176] Compound 10-5 (25.13 g, 0.1 mol) was dissolved in 150 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-5 with a yield of 92.0% and a purity of 99.92%.

[0177] Preparation of compound TM5:

[0178] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-5 (18.14 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM5 with a yield of 91.9% and a purity of 99.89%.

[0179] 1 HNMR(400MHz,DMSO-d6)δ12.15(s,1H),9.76(s,1H),8.98(s,1H),7.82~7. 74(m,3H),7.33~7.23(m,3H),7.11~7.04(m,1H),4.65(s,2H),3.42(s,2H);

[0180] 13C-NMR(100MHz,DMSO-d6):168.4,162.9,157.6,155.5,150.2,141.1,137.1,133.4 ,134.3,130.4,129.5,128.8,127.5,124.8,121.8,118.7,112.2,98.7,48.8,30.5.

[0181] MS (m / z): 501.16 [M+H] + ;

[0182] Example 6

[0183] Preparation of compound TM6:

[0184]

[0185] Preparation of compound 10-6:

[0186] Compound 8 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-6 (16.46 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-6 with a yield of 92.2% and a purity of 99.84%.

[0187] Preparation of compound 11-6:

[0188] Compound 10-6 (29.44 g, 0.1 mol) was dissolved in 150 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-6 with a yield of 92.2% and a purity of 99.91%.

[0189] Preparation of compound TM6:

[0190] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-6 (23.71 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM6 with a yield of 92.4% and a purity of 99.91%.

[0191] 1HNMR(400MHz,DMSO-d6)δ12.15(s,1H),9.76(s,1H),8.98(s,1H),7.82~7.74(m,3H), 7.63(s,1H),7.23~7.33(m,2H),4.92(s,2H),3.57(s,2H),3.32(s,3H),2.15(s,3H);

[0192] 13 C-NMR(100MHz,DMSO-d6):168.4,167.9,157.6,155.5,150.2,141.1,135.1,133.4,132.5,1 31.0,129.5,128.1,125.4,123.3,122.8,120.9,118.7,112.2,98.7,56.4,54.8,27.0,18.2.

[0193] MS (m / z): 544.22 [M+H] + ;

[0194] Example 7

[0195] Preparation of compound TM7:

[0196]

[0197] Preparation of compound 10-7:

[0198] Compound 8 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-7 (15.98 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-7 with a yield of 92.0% and a purity of 99.82%.

[0199] Preparation of compound 11-7:

[0200] Compound 10-7 (29.03 g, 0.1 mol) was dissolved in 150 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The organic layer was washed with purified water and dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-7 with a yield of 91.8% and a purity of 99.88%.

[0201] Preparation of compound TM7:

[0202] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-7 (22.82 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM7 with a yield of 92.0% and a purity of 99.87%.

[0203] 1 HNMR (400MHz, DMSO-d6) δ12.15(s,1H),10.56(s,1H),9.76(s,1H),8.98(s,1H),7.82~7.74(m,3H),7.23~7.13(m,4H),4.65(s,2H),3.72(s,2H);

[0204] 13 C-NMR(100MHz,DMSO-d6):168.4,157.6,155.5,150.2,141.1,136.2(2C),133.4,130. 3,129.5,126.8,123.2(2C),122.5,120.9,118.7,115.2(2C),112.2,98.7,54.8,27.2.

[0205] MS (m / z): 540.13 [M+H] + ;

[0206] Example 8

[0207] Preparation of compound TM8:

[0208]

[0209] Preparation of compound 10-8:

[0210] Compound 8 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-8 (32.93 g, 0.12 mol), HOBt (13.51 g, 0.1 mol) and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-8 with a yield of 92.5% and a purity of 99.87%.

[0211] Preparation of compound 11-8:

[0212] Compound 10-8 (43.16 g, 0.1 mol) was dissolved in 150 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-8 with a yield of 92.4% and a purity of 99.90%.

[0213] Preparation of compound TM8:

[0214] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-8 (39.78 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM8 with a yield of 92.3% and a purity of 99.90%.

[0215] 1 HNMR(400MHz,DMSO-d6)δ12.15(s,1H),9.76(s,1H),8.98(s,1H),7.82~7.74(m,4H),7.23~7.33(m ,3H),4.72(s,2H),3.49(s,2H),3.11(m,4H);2.74(m,1H);2.45(m,8H);2.36(s,3H);1.82(m,4H);

[0216] 13 C-NMR(100MHz,DMSO-d6):168.4,167.2,157.6,155.5,150.2,141.1,133.4,132.7,129.5,120.9,120.6(2C),1 18.7,118.4,118.3,117.7(2C),112.2,98.7,63.4(2C),56.7(2C),54.8,54.3(2C),53.3,45.3,29.4(2C),27.0.

[0217] MS (m / z): 681.16 [M+H]+;

[0218] Example 9

[0219] Preparation of compound TM9:

[0220]

[0221] Preparation of compound 10-9:

[0222] Compound 8 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-9 (16.46 g, 0.12 mol), HOBt (13.51 g, 0.1 mol) and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-9 with a yield of 92.1% and a purity of 99.85%.

[0223] Preparation of compound 11-8:

[0224] Compound 10-9 (19.44 g, 0.1 mol) was dissolved in 150 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-9 with a yield of 92.0% and a purity of 99.87%.

[0225] Preparation of compound TM9:

[0226] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-9 (23.31 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM9 with a yield of 92.0% and a purity of 99.89%.

[0227] MS (m / z): 544.15 [M+H] + ; 1 H-NMR (400MHz, DMSO-d6): δ11.08(s,1H),10.24(s,1H),8.70(s,1H),7.55(s,1H),7.28~7 .16(m,5H),4.83~4.64(m,2H),3.77(s,3H),3.46(s,2H),2.68~2.54(m,2H);2.18(s,3H); 13 C-NMR (400MHz, DMSO-d6): δ173.0,168.8,167.9,162.4,158.9,149.8,137.7,135.5,130.8,128. 3,125.7,123.0,122.5,121.5,119.4,118.9,118.6,115.3,114.6,72.2,56.6,39.7,35.5,19.9.

[0228] Example 10

[0229] Preparation of compound TM10:

[0230]

[0231] Preparation of compound 10-10:

[0232] Compound 8-1 (23.73 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-10 (16.46 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-10 with a yield of 91.7% and a purity of 99.82%.

[0233] Preparation of compound 11-10:

[0234] Compound 10-10 (35.64 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-10 with a yield of 92.0% and a purity of 99.87%.

[0235] Preparation of compound TM10:

[0236] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-10 (30.76 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM10 with a yield of 92.2% and a purity of 99.91%.

[0237] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),7.71~7.54 (m,4H),7.45(s,1H),7.25~7.17(m,5H),3.71(s,3H),3.59(s,2H),2.15(s,3H);

[0238] 13C-NMR (400MHz, DMSO-d6): δ172.8,168.8,167.7,162.8,158.6,149.7,142.4,137.5,135.7,131.2(2C),130.4 ,128.5,125.8,124.3,123.4,122.3,121.2,119.5,118.7,118.4,115.1,114.8,113.5(2C),72.4,56.8,19.7.

[0239] MS (m / z): 606.20 [M+H] + ;

[0240] Embodiment 11

[0241] Preparation of compound TM11:

[0242]

[0243] Preparation of compound 10-11:

[0244] Compound 8-1 (23.73 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-11 (8.90 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-11 with a yield of 92.0% and a purity of 99.85%.

[0245] Preparation of compound 11-11:

[0246] Compound 10-11 (29.34 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-11 with a yield of 92.4% and a purity of 99.89%.

[0247] Preparation of compound TM11:

[0248] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-11 (23.19 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM11 with a yield of 92.0% and a purity of 99.88%.

[0249] 1H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),7.71~7.54 (m,3H),7.75(s,1H),7.25~7.17(m,3H),4.15(s,2H),3.59(s,2H),2.33(s,6H);

[0250] 13 C-NMR (400MHz, DMSO-d6): δ168.8,167.7,162.8,158.6,149.7,142.4,137.5,131.2,130. 4,128.5(2C),125.8(2C),123.4,121.2,119.5,114.8,110.5,99.5,72.4,47.4(2C),31.2.

[0251] MS (m / z): 543.13 [M+H] + ;

[0252] Example 12

[0253] Preparation of compound TM12:

[0254]

[0255] Preparation of compound 10-12:

[0256] Compound 8-1 (23.73 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-12 (8.90 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-12 with a yield of 92.0% and a purity of 99.85%.

[0257] Preparation of compound 11-12:

[0258] Compound 10-12 (29.34 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-12 with a yield of 92.4% and a purity of 99.89%.

[0259] Preparation of compound TM12:

[0260] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-12 (23.19 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM12 with a yield of 92.0% and a purity of 99.88%.

[0261] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),7.71~7.54(m,3H ),7.75(s,1H),7.25~7.17(m,3H),3.59(s,2H),3.49~3.41(m,1H),1.59~1.22(m,10H);

[0262] 13 C-NMR (400MHz, DMSO-d6): δ168.8,167.7,162.8,158.6,149.7,142.4,137.5,131.2,130.4,128.5 (2C),125.8(2C),123.4,121.2,119.5,114.8,110.5,99.5,55.4,33.4(2C),31.2,26.8(2C),23.9.

[0263] MS (m / z): 568.09 [M+H] + ;

[0264] Embodiment 13

[0265] Preparation of compound TM13:

[0266]

[0267] Preparation of compound 10-13:

[0268] Compound 8-1 (23.73 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-13 (11.18 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-13 with a yield of 91.8% and a purity of 99.84%.

[0269] Preparation of compound 11-13:

[0270] Compound 10-13 (31.24 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The organic layer was washed with purified water and dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-13 with a yield of 92.6% and a purity of 99.91%.

[0271] Preparation of compound TM13:

[0272] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-13 (25.47 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM13 with a yield of 91.7% and a purity of 99.82%.

[0273] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),7.75(s ,1H),7.71~7.54(m,5H),7.25~7.17(m,5H),7.15~7.07(m,1H),3.59(s,2H);

[0274] 13 C-NMR (400MHz, DMSO-d6): δ168.8,167.7,162.8,158.6,149.7,142.4,137.5,136.7,131.2,130.4,1 29.1(2C),128.5(2C),127.4(2C),125.8(2C),123.4,122.5,121.2,119.5,114.8,110.5,99.5,31.2.

[0275] MS (m / z): 562.21 [M+H] + ;

[0276] Embodiment 14

[0277] Preparation of compound TM14:

[0278]

[0279] Preparation of compound 10-14:

[0280] Compound 8-1 (23.73 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-14 (13.33 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-14 with a yield of 91.4% and a purity of 99.81%.

[0281] Preparation of compounds 11-14:

[0282] Compound 10-14 (33.04 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-14 with a yield of 92.0% and a purity of 99.87%.

[0283] Preparation of compound TM14:

[0284] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-14 (27.63 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM14 with a yield of 91.1% and a purity of 99.80%.

[0285] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),7.99~7.75(m,2H),7.71~7.54(m,4H),7.25~7.17(m,5H),3.59(s,2H);

[0286] 13 C-NMR (400MHz, DMSO-d6): δ168.8,167.7,162.8,161.5,158.6,149.7,142.4,137.5,136.7,133.4,1 30.4,128.5(2C),125.8(2C),127.7(2C),123.4,122.5,121.2,119.5,117.1(2C),110.5,99.5,31.2.

[0287] MS (m / z): 580.15 [M+H] + ;

[0288] Embodiment 15

[0289] Preparation of compound TM15:

[0290]

[0291] Preparation of compound 10-15:

[0292] Compound 8-1 (23.73 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-15 (11.29 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-15 with a yield of 92.0% and a purity of 99.87%.

[0293] Preparation of compound 11-15:

[0294] Compound 10-15 (31.34 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The organic layer was washed with purified water and dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-15 with a yield of 92.3% and a purity of 99.89%.

[0295] Preparation of compound TM15:

[0296] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-15 (25.59 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM15 with a yield of 91.9% and a purity of 99.88%.

[0297] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.03~7.92(m,3H),7.81~7.70(m,3H),7.25~7.17(m,5H),3.59(s,2H);

[0298] 13C-NMR (400MHz, DMSO-d6): δ168.8,167.7,162.8,158.6,154.3,149.7,146.8,142.4,139.4,137.5, 136.7,133.4,130.4,128.5(2C),125.8(2C),123.4,122.5,121.2,119.5,117.9,113.5,99.5,31.2.

[0299] MS (m / z): 563.11 [M+H] + ;

[0300] Example 16

[0301] Preparation of compound TM16:

[0302]

[0303] Preparation of compound 10-16:

[0304] Compound 8-1 (23.73 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-16 (17.18 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-16 with a yield of 91.5% and a purity of 99.84%.

[0305] Preparation of compound 11-16:

[0306] Compound 10-16 (36.24 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-16 with a yield of 91.8% and a purity of 99.82%.

[0307] Preparation of compound TM16:

[0308] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-16 (31.48 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM16 with a yield of 91.4% and a purity of 99.85%.

[0309] 1H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.03~7.9 2(m,2H),7.85~7.71(m,4H),7.57~7.41(m,4H),7.25~7.17(m,4H),3.59(s,2H);

[0310] 13 C-NMR (400MHz, DMSO-d6): δ168.8,167.7,162.8,158.6,154.3,149.7,146.8,142.4,139.4,137.5,135.7,133.4,132 .8,130.4,129.1,128.5(2C),126.7,125.8(2C),124.3,122.5,120.5,119.5,119.1,117.9,116.7,113.5,99.5,31.2.

[0311] MS (m / z): 612.16 [M+H] + ;

[0312] Embodiment 17

[0313] Preparation of compound TM17:

[0314]

[0315] Preparation of compound 10-17:

[0316] Compound 8-1 (23.73 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-17 (15.98 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-17 with a yield of 91.9% and a purity of 99.86%.

[0317] Preparation of compound 11-17:

[0318] Compound 10-17 (33.52 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The organic layer was washed with purified water and dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-17 with a yield of 92.1% and a purity of 99.89%.

[0319] Preparation of compound TM17:

[0320] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-17 (30.27 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM17 with a yield of 91.7% and a purity of 99.86%.

[0321] 1 H-NMR (400MHz, DMSO-d6): δ12.11(s,1H),11.03(s,1H),10.20(s,1H),8.74(s,1H), 7.71~7.54(m,3H),7.75(s,1H),7.25~7.17(m,3H),7.11~7.04(m,4H),3.59(s,2H);

[0322] 13 C-NMR (400MHz, DMSO-d6): δ168.8,167.7,162.8,158.6,150.7,149.7,142.4,139.1,137.5,136.2,131. 2,130.4,128.5(2C),125.8(2C),124.6(2C),123.4,121.2,119.5,117.0(2C),114.8,110.5,99.5,31.2;

[0323] MS (m / z): 602.18 [M+H] + ;

[0324] Embodiment 18

[0325] Preparation of compound TM18:

[0326]

[0327] Preparation of compound 10-18:

[0328] Compound 8-1 (23.73 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-18 (32.93 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-18 with a yield of 92.4% and a purity of 99.88%.

[0329] Preparation of compound 11-18:

[0330] Compound 10-18 (49.37 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-18 with a yield of 92.3% and a purity of 99.92%.

[0331] Preparation of compound TM18:

[0332] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-18 (47.22 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM18 with a yield of 92.1% and a purity of 99.88%.

[0333] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),7.71~7.54(m,3H),7.75(s,1H),7.31~7.22(m ,5H),7.11~6.94(m,2H),3.59(s,2H),3.34~7.21(m,4H),2.77~2.63(m,1H),2.36~2.18(m,11H),1.77~1.65(m,4H);

[0334] 13 C-NMR (400MHz, DMSO-d6): δ168.8,167.7,162.8,158.6,149.7,145.7,142.4,137.5,131.2,130.4,129.4,128.5(2C),125.8(2C ),123.4,122.6(2C),121.2,119.5,115.2(2C),114.8,110.5,99.5,76.2,66.1(2C),61.5(2C),55.3(2C),47.6,31.2,29.3(2C).

[0335] MS (m / z): 743.24 [M+H] + ;

[0336] Embodiment 19

[0337] Preparation of compound TM19:

[0338]

[0339] Preparation of compound 10-19:

[0340] Compound 8-1 (23.73 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-19 (17.79 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-19 with a yield of 91.7% and a purity of 99.80%.

[0341] Preparation of compound 11-19:

[0342] Compound 10-19 (36.85 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-19 with a yield of 91.8% and a purity of 99.89%.

[0343] Preparation of compound TM19:

[0344] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-19 (32.20 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM19 with a yield of 91.6% and a purity of 99.84%.

[0345] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),7.84~7.75(m,2H),7.70~7.56(m,5H),7.35~7.20(m,5H),3.59(s,2H);

[0346] 13 C-NMR (400MHz, DMSO-d6): δ168.8,167.7,162.8,158.6,149.7,142.4,139.2,138.4,137.5,131.2,130.4,128 .5(2C),127.3,125.8(2C),124.6,124.3,123.4,123.0,122.5,121.2,119.5,114.8,111.0,110.5,99.5,31.2.

[0347] MS (m / z): 618.19 [M+H] + ;

[0348] Embodiment 20

[0349] Preparation of compound TM20:

[0350]

[0351] Preparation of compound 10-20:

[0352] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-20 (8.90 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-20 with a yield of 91.1% and a purity of 99.75%.

[0353] Preparation of compound 11-20:

[0354] Compound 10-20 (24.53 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-20 with a yield of 91.2% and a purity of 99.83%.

[0355] Preparation of compound TM20:

[0356] The intermediate 6 (18.41 g, 0.05 mol) was added to a 100 ml three-necked flask, 80 ml of toluene was poured in, and compound 11-20 (17.43 g, 0.12 mol) was added, and the mixture was heated to reflux for reaction. The reaction was completed by TLC detection, and the mixture was cooled to 0°C for crystallization. After the crystallization was completed, the mixture was filtered, and the filter cake was rinsed with an appropriate amount of toluene and dried under reduced pressure to obtain compound TM20 with a yield of 91.0% and a purity of 99.80%.

[0357] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.11~7.98(m,2H),7.2 5~7.17(m,1H),4.88~4.81(m,2H),4.31(s,2H),3.59(s,2H),2.71~2.64(m,2H),2.26(s,6H);

[0358] 13C-NMR (400MHz, DMSO-d6): δ170.1,168.6,167.8,159.2,149.7,145.6,142.4 ,133.1,132.3,128.2,118.5,111.0,99.6,79.2,47.2(2C),40.1,33.7,31.2.

[0359] MS (m / z): 495.32 [M+H] + ;

[0360] Embodiment 21

[0361] Preparation of compound TM21:

[0362]

[0363] Preparation of compound 10-21:

[0364] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-21 (11.90 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-21 with a yield of 91.4% and a purity of 99.78%.

[0365] Preparation of compound 11-21:

[0366] Compound 10-21 (27.04 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-21 with a yield of 91.9% and a purity of 99.88%.

[0367] Preparation of compound TM21:

[0368] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-21 (20.43 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM21 with a yield of 92.1% and a purity of 99.89%.

[0369] 1H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.11~7.98(m,2H),7.25~7.17(m,1H),4.31(s ,2H),3.59(s,2H),3.51~3.42(m,1H),2.71~2.64(m,2H),1.74~1.69(m,2H),1.50~1.43(m,4H),1.23~1.11(m,4H);

[0370] 13 C-NMR (400MHz, DMSO-d6): δ170.1,168.6,167.8,159.2,149.7,145.6,142.4,133.1, 132.3,128.2,118.5,111.0,99.6,50.5,40.1,35.6(2C),33.7,31.2,25.6,26.7(2C).

[0371] MS (m / z): 520.11 [M+H] + ;

[0372] Embodiment 22

[0373] Preparation of compound TM22:

[0374]

[0375] Preparation of compound 10-22:

[0376] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-22 (11.18 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-22 with a yield of 91.6% and a purity of 99.79%.

[0377] Preparation of compound 11-22:

[0378] Compound 10-22 (26.43 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-22 with a yield of 91.1% and a purity of 99.82%.

[0379] Preparation of compound TM22:

[0380] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-22 (19.71 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, rinse the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM22 with a yield of 91.5% and a purity of 99.82%.

[0381] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.11~7 .98(m,2H),7.25~7.17(m,6H),4.31(s,2H),3.59(s,2H),2.71~2.64(m,2H);

[0382] 13 C-NMR (400MHz, DMSO-d6): δ170.1,168.6,167.8,159.2,149.7,145.6,142.4,139.5,13 3.1,132.3,128.8(2C),128.2,127.1,121.8(2C),118.5,111.0,99.6,40.1,33.7,31.2.

[0383] MS (m / z): 514.17 [M+H] + ;

[0384] Embodiment 23

[0385] Preparation of compound TM23:

[0386]

[0387] Preparation of compound 10-23:

[0388] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-23 (13.33 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-23 with a yield of 91.0% and a purity of 99.74%.

[0389] Preparation of compound 11-23:

[0390] Compound 10-23 (28.23 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-23 with a yield of 91.4% and a purity of 99.83%.

[0391] Preparation of compound TM23:

[0392] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-23 (21.86 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM23 with a yield of 91.7% and a purity of 99.84%.

[0393] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.11~7 .98(m,2H),7.25~7.17(m,5H),4.31(s,2H),3.59(s,2H),2.71~2.64(m,2H);

[0394] 13 C-NMR (400MHz, DMSO-d6): δ170.1,168.6,167.8,162.3,159.2,149.7,145.6,142.4,13 3.1,132.3,131.5,128.2,121.8(2C),118.5,115.2(2C),111.0,99.6,40.1,33.7,31.2.

[0395] MS (m / z): 532.24 [M+H] + ;

[0396] Embodiment 24

[0397] Preparation of compound TM24:

[0398]

[0399] Preparation of compound 10-24:

[0400] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-24 (11.29 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-24 with a yield of 91.6% and a purity of 99.77%.

[0401] Preparation of compound 11-24:

[0402] Compound 10-24 (26.53 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-24 with a yield of 91.8% and a purity of 99.85%.

[0403] Preparation of compound TM24:

[0404] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-24 (19.82 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM24 with a yield of 91.8% and a purity of 99.87%.

[0405] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.11~7.98(m,2H ),7.33~7.19(m,4H),7.11~7.04(m,1H),4.31(s,2H),3.59(s,2H),2.71~2.64(m,2H);

[0406] 13 C-NMR (400MHz, DMSO-d6): δ170.1,168.6,167.8,159.2,151.3,149.7,147.3,145.6,1 42.4,139.2,133.1,132.3,128.2,124.5,118.5,115.6,111.0,99.6,40.1,33.7,31.2.

[0407] MS (m / z): 515.19 [M+H] + ;

[0408] Embodiment 25

[0409] Preparation of compound TM25:

[0410]

[0411] Preparation of compound 10-25:

[0412] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-25 (16.46 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-25 with a yield of 91.0% and a purity of 99.71%.

[0413] Preparation of compound 11-25:

[0414] Compound 10-25 (30.84 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The organic layer was washed with purified water and dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-25 with a yield of 91.3% and a purity of 99.83%.

[0415] Preparation of compound TM25:

[0416] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-25 (24.99 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM25 with a yield of 91.0% and a purity of 99.81%.

[0417] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.11~7.98(m,2H),7.82~7.74 (m,3H),7.25~7.17(m,1H),4.31(s,2H),3.59(s,2H),3.32(s,3H),2.71~2.64(m,2H),2.15(s,3H);

[0418] 13C-NMR (400MHz, DMSO-d6): δ170.1,168.6,167.8,159.2,157.4,149.7,145.6,142.4,137.3,133 .1,132.3,130.3,128.2,122.1,118.5,114.2,111.0,103.9,99.6,55.6,40.1,33.7,31.2,15.9.

[0419] MS (m / z): 558.07 [M+H] + ;

[0420] Embodiment 26

[0421] Preparation of compound TM26:

[0422]

[0423] Preparation of compound 10-26:

[0424] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-26 (15.98 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-26 with a yield of 91.9% and a purity of 99.82%.

[0425] Preparation of compound 11-26:

[0426] Compound 10-26 (30.44 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-26 with a yield of 91.9% and a purity of 99.87%.

[0427] Preparation of compound TM26:

[0428] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-26 (24.51 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM26 with a yield of 92.2% and a purity of 99.90%.

[0429] 1H-NMR (400MHz, DMSO-d6): δ12.15(s,1H),11.03(s,1H),10.20(s,1H),8.74(s,1H), 8.11~7.98(m,2H),7.25~7.17(m,5H),4.31(s,2H),3.59(s,2H),2.71~2.64(m,2H);

[0430] 13 C-NMR (400MHz, DMSO-d6): δ170.1,168.6,167.8,159.2,149.7,146.5,145.6,142.4,137.2 ,136.4,133.1,132.3,128.2,123.2(2C),118.5,115.2(2C),111.0,99.6,40.1,33.7,31.2.

[0431] MS (m / z): 554.30 [M+H] + ;

[0432] Embodiment 27

[0433] Preparation of compound TM27:

[0434]

[0435] Preparation of compound 10-27:

[0436] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-27 (32.93 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-27 with a yield of 92.3% and a purity of 99.87%.

[0437] Preparation of compound 11-27:

[0438] Compound 10-27 (46.17 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-27 with a yield of 92.1% and a purity of 99.89%.

[0439] Preparation of compound TM27:

[0440] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-27 (41.46 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM27 with a yield of 92.4% and a purity of 99.91%.

[0441] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.11~7.98(m,2H),7.82~7.74(m,4H),7.25~7 .17(m,1H),4.31(s,2H),3.59(s,2H),3.34~7.21(m,4H),2.77~2.63(m,3H),2.36~2.18(m,11H),1.77~1.65(m,4H);

[0442] 13 C-NMR (400MHz, DMSO-d6): δ170.1,168.6,167.8,159.2,149.7,146.6,145.6,142.4,133.1,132.3,128.2,127.3,1 26.1(2C),118.5,113.4(2C),111.0,99.6,63.4(2C),70.5,56.7(2C),54.3(2C),46.8,40.1,33.7,31.2,29.4(2C).

[0443] MS (m / z): 695.27 [M+H] + ;

[0444] Embodiment 28

[0445] Preparation of compound TM28:

[0446]

[0447] Preparation of compound 10-28:

[0448] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-28 (18.02 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-28 with a yield of 91.5% and a purity of 99.82%.

[0449] Preparation of compound 11-28:

[0450] Compound 10-28 (32.14 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-28 with a yield of 91.1% and a purity of 99.80%.

[0451] Preparation of compound TM28:

[0452] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-28 (26.55 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM28 with a yield of 91.8% and a purity of 99.87%.

[0453] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.18~7.96(m,4H ),7.62~7.54(m,2H),7.25~7.17(m,1H),4.31(s,2H),3.59(s,2H),2.71~2.64(m,2H);

[0454] 13 C-NMR (400MHz, DMSO-d6): δ174.0,170.1,168.6,167.8,159.2,155.2,149.7,145.6,142.4,1 33.1,132.3,131.2,128.2,122.5,123.6,120.3,119.1,118.5,111.0,99.6,40.1,33.7,31.2.

[0455] MS (m / z): 571.22 [M+H] + ;

[0456] Embodiment 29

[0457] Preparation of compound TM29:

[0458]

[0459] Preparation of compound 10-29:

[0460] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-29 (17.18 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-29 with a yield of 91.2% and a purity of 99.81%.

[0461] Preparation of compound 11-29:

[0462] Compound 10-29 (31.44 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-29 with a yield of 91.4% and a purity of 99.83%.

[0463] Preparation of compound TM29:

[0464] Add intermediate 6 (18.41 g, 0.05 mol) into a 100 ml three-necked flask, pour in 80 ml of toluene, add compound 11-29 (25.71 g, 0.12 mol), heat to reflux for reaction, detect the completion of the reaction by TLC, cool to 0°C for crystallization, filter, wash the filter cake with an appropriate amount of toluene, and dry under reduced pressure to obtain compound TM29 with a yield of 91.4% and a purity of 99.85%.

[0465] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.38~8.32(m,1H),8.11~7 .98(m,4H),7.62~7.47(m,3H),7.25~7.18(m,2H),4.31(s,2H),3.59(s,2H),2.71~2.64(m,2H);

[0466] 13 C-NMR (400MHz, DMSO-d6): δ170.1,168.6,167.8,159.2,149.7,145.6,142.4,136.2,134.7,133.1,132. 3,129.5,128.2,127.0,126.6,124.3,122.7,121.3,119.5,118.5,117.1,111.0,99.6,40.1,33.7,31.2.

[0467] MS (m / z): 564.16 [M+H] + ;

[0468] Embodiment 30

[0469] Preparation of compound TM30:

[0470]

[0471] Preparation of compound 10-30:

[0472] Compound 8-2 (18.92 g, 0.1 mol) was added to dried dichloromethane, followed by compound 9-30 (15.31 g, 0.12 mol), HOBt (13.51 g, 0.1 mol), and EDCI (19.17 g, 0.1 mol), stirred at room temperature, and tested by TLC. After the reaction was completed, the mixture was evaporated to dryness under reduced pressure and separated by column chromatography to obtain compound 10-30 with a yield of 91.6% and a purity of 99.83%.

[0473] Preparation of compound 11-30:

[0474] Compound 10-30 (29.88 g, 0.1 mol) was dissolved in 180 ml of ethyl acetate, and 30 mL of 3 mol / L hydrochloric acid was added dropwise to remove the Boc group. The mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness under reduced pressure to obtain compound 11-30 with a yield of 91.9% and a purity of 99.88%.

[0475] Preparation of compound TM30:

[0476] The intermediate 6 (18.41 g, 0.05 mol) was added to a 100 ml three-necked flask, 80 ml of toluene was poured in, and compound 11-30 (23.84 g, 0.12 mol) was added, and the mixture was heated to reflux for reaction. The reaction was completed by TLC detection, and the mixture was cooled to 0°C for crystallization. After the crystallization was completed, the mixture was filtered, and the filter cake was rinsed with an appropriate amount of toluene and dried under reduced pressure to obtain compound TM30 with a yield of 92.0% and a purity of 99.89%.

[0477] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.11~7.98(m,2H ),7.53~7.40(m,4H),7.25~7.17(m,1H),4.31(s,2H),3.59(s,2H),2.71~2.64(m,2H);

[0478] 13C-NMR (400MHz, DMSO-d6): δ170.1,168.6,167.8,159.2,149.7,145.6,142.4,136.5,13 4.2,133.1,132.3,130.1(2C),128.2,119.3(2C),118.5,111.0,99.6,40.1,33.7,31.2.

[0479] MS (m / z): 548.27 [M+H] + ;

[0480] In vitro cytotoxic activity test against cancer cells

[0481] In order to study the ability of the target compound synthesized in this experiment to inhibit tumor cell proliferation, we determined the in vitro cytotoxicity of the compound of the present invention on four tumor cells, namely, lymphoma cell line (Raji), breast cancer cell (MCF7), leukemia cell (K562) and head and neck squamous cell carcinoma cell (HN5). The detection method used in the experiment was the standard MTT method.

[0482] The specific experimental method is:

[0483] Lymphoma cell line, breast cancer cell, leukemia cell, head and neck squamous cell carcinoma cell (Shanghai Hexu Biotechnology Co., Ltd.) four kinds of tumor cells were digested with trypsin for 10 minutes, and then the liquid was discarded. The cells were blown with 5% serum culture medium to adjust the cell concentration to 300-400 / μL, and the drugs were added in sequence. The blank group containing only culture medium was reserved and cultured in the incubator for 24 hours. The supernatant was discarded, and the diluted target compound of different concentrations was added to a 96-well plate (Shanghai Wohong Biotechnology Co., Ltd.). The control group only added cell solution. Three sub-wells were set for each concentration. After mixing, the culture was continued for 36 hours. The cell morphology changes at different time periods were observed. After the cells and the drug were fully reacted, the freshly prepared MTT solution was added to each well for color development, and the culture was continued for 6 hours. The upper liquid was discarded, 100μLDMSO was dissolved to form purple crystals, and 490nm was selected. The absorbance was measured by an enzyme reader (HBS-1096A enzyme label analyzer, Nanjing Detie Experimental Equipment Co., Ltd.). IC was calculated by the formula: cell inhibition rate (%) = (1-A experimental / A control) × 100%. 50 Values ​​(using cell survival rate versus log dose plot method).

[0484] The results of in vitro toxicity tests of compounds TM1-8 on the proliferation inhibition of four tumor cells, namely lymphoma cell line (Raji), breast cancer cell (MCF7), leukemia cell (K562) and head and neck squamous cell carcinoma cell (HN5), are shown in Table 1.

[0485] Table 1 Toxicity test of some compounds on tumor cells (IC 50,μM) a

[0486]

[0487]

[0488] Note: a The values ​​in the table are the average values ​​of 3 experiments; b The drug action time is 36 hours

[0489] The novel anti-tumor active compounds TM1-30 provided by the present invention have high growth inhibitory activities on four tumor cells, namely, lymphoma cell line (Raji), breast cancer cell (MCF7), leukemia cell (K562) and head and neck squamous cell carcinoma cell (HN5), while the inhibition rates of compound TM10 on lymphoma cell line (Raji) and leukemia cell (K562) both reach nM level; the inhibition rate of TM19 on MCF7 reaches nM level; and the inhibition rates of TM26 on Raji and MCF7 both reach nM level; and the inhibition rates of the three compounds on the four tumor cells are significantly higher than those of the control compound ibrutinib; the inhibitory activity and selectivity of compound TM25 on MCF7 cells are better than those of the other three cells.

[0490] The compound TM10 of the present invention also exhibits good physical properties, with a water solubility of 0.115 mg / ml; meanwhile, it is well metabolized in mice, with a bioavailability of 96.8%.

Claims

1. A pyrazole-4,6-dione compound, characterized in that: Has the following structure:

2. A compound I The preparation method is characterized in that The following steps are involved: Preparation of compound 6: General synthesis method of compound 11: Preparation of Compound I: Wherein, R is independently selected from H, halogen, aryl, heteroaryl, C 1-4 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 2-6 heterocycloalkyl, aryloxy, -NR2R3; or substituted aryl, substituted heteroaryl, substituted C 3-6 Cycloalkyl, substituted C 2-6 Heterocycloalkyl, substituted aryloxy, substituted C 1-6 One or more independent groups in an alkoxy group, wherein the substituent group is selected from halogen, aryl, heteroaryl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 1-4 Alkyl, substituted C 2-6 Heterocycloalkyl, halogen-substituted C 2-6 Heterocycloalkyl, C 2-6 One or more substituents in the heterocycloalkyl group. R2 is independently selected from H or C 1-4 alkyl; R3 is independently selected from H, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-3 Haloalkyl, aryl, heteroaryl, C 1-6 Alkoxy, C 1-6 Alkylamino, substituted C 2-6 Heterocycloalkyl, halogen-substituted C 2-6 Heterocycloalkyl, C 2-6 one or more of heterocycloalkyl; Z is independently selected from C 1-4 Alkyl or aryl.

3. The method for preparing the pyrazole-4,6-dione compound according to claim 2, characterized in that: The reaction steps include: Step 1: Compound 1 undergoes condensation reaction in sodium ethoxide solution to obtain compound 2; Step 2: Compound 2 reacts with hydrazine hydrate to obtain compound 3; Step 3: Compound 3 reacts under alkaline conditions to obtain compound 4; Step 4: Compound 4 is subjected to temperature-controlled reflux dehydration to generate intermediate compound 5; Step 5: Compound 5 reacts with 3-bromo-4-fluorobenzoic acid, EDCI and HOBt in dichloromethane to obtain compound 6; Step 6: Compound 8 and compound 9 react to obtain compound 10; Step 7: Compound 10 is hydrolyzed to obtain compound 11; Step 8: Compound 6 and compound 11 are reacted in an organic solvent, and the target compound I is obtained after crystallization, filtration and drying.

4. The method for preparing the pyrazole-4,6-dione compound according to claim 2, characterized in that: The specific steps of the reaction include: Step 1: Add sodium ethoxide to ethanol solvent, stir to dissolve, add compound 1, raise the temperature to reflux, detect by TLC, after the reaction is complete, cool down to crystallize, filter by suction, and dry to obtain compound 2; Step 2: Add compound 2 to water, stir to dissolve, add hydrazine hydrate, raise the temperature to reflux, detect by TLC, after the reaction is complete, cool down to precipitate solid, filter, and dry to obtain compound 3; Step 3: Compound 3 is added to the alkaline solution, stirred and dissolved, heated to reflux, and detected by TLC. After the reaction is completed, the temperature is lowered and hydrochloric acid is added to adjust the pH to 3-4, and the solid is precipitated, filtered, and dried to obtain compound 4; Step 4: dissolving compound 4 in acetic anhydride solution, adding a condensing agent, heating to reflux, and performing TLC detection. After the reaction is completed, cooling and crystallizing, filtering, rinsing the filter cake with methyl tert-butyl ether, and drying to obtain compound 5; Step 5: Dissolve 3-bromo-4-fluorobenzoic acid in dried dichloromethane, add EDCI, HOBt and compound 5 in sequence, stir to react at room temperature, detect by TLC, and after the reaction is complete, filter, concentrate, evaporate to dryness under reduced pressure, and separate by column chromatography to obtain compound 6; Step 6: Compound 8 is added to dried dichloromethane, and compound 9 and a condensing agent are added in sequence, stirred at room temperature, and tested by TLC. After the reaction is completed, the mixture is evaporated to dryness under reduced pressure, and separated by column chromatography to obtain compound 10; Step 7: Dissolve compound 10 in an organic solvent, add hydrochloric acid dropwise to remove the Boc group, extract with an extractant, dry with anhydrous sodium sulfate, filter, concentrate, and evaporate to dryness under reduced pressure to obtain compound 11; Step 8: Compound 6 is dissolved in an organic solvent, compound 11 is added, and the mixture is heated to reflux for reaction. The reaction is completed by TLC detection. The mixture is cooled for crystallization, the filter cake is washed with an organic solvent, and dried to obtain compound I.

5. The method for preparing the pyrazole-4,6-dione compound according to claim 4, characterized in that: The base described in step 3 is one or two of sodium hydride, potassium hydroxide, sodium hydroxide and lithium hydroxide.

6. The method for preparing the pyrazole-4,6-dione compound according to claim 4, characterized in that: The condensing agent in step 4 is one or two of dicyclohexylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, and 2-chloro-1,3-dimethylimidazoline chloride.

7. The method for preparing the pyrazole-4,6-dione compound according to claim 4, characterized in that: The condensation agent in step 6 is one or two of dicyclohexylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 2-chloro-1,3-dimethylimidazoline chloride, and 1-carbonylbenzotriazole.

8. The method for preparing pyrazole-4,6-dione compounds according to claim 4, characterized in that: The organic solvent described in step 7 is one of ethyl acetate, dichloromethane, methanol and ethanol.

9. The method for preparing the pyrazole-4,6-dione compound according to claim 4, characterized in that: The organic solvent described in step 8 is one of toluene, benzene and xylene.

10. A pharmaceutical composition comprising the pyrazole-4,6-dione compound according to claim 1, characterized in that: The pharmaceutical composition comprises: a therapeutically effective amount of the pyrazole-4,6-dione compound according to claim 1 and an optional pharmaceutically acceptable carrier, excipient, adjuvant, auxiliary material or diluent.

11. A use of the pyrazole-4,6-dione compound as claimed in claim 1, characterized in that: The invention is used as an active ingredient in preparing medicines for treating lymphoma, breast cancer, leukemia and head and neck squamous cell carcinoma.

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