CN114057724B

CN114057724B - A BTK inhibitor

Info

Publication number: CN114057724B
Application number: CN202010743104.7A
Authority: CN
Inventors: 张贵民; 唐贞波; 张永波
Original assignee: Lunan Pharmaceutical Group Corp
Current assignee: Lunan Pharmaceutical Group Corp
Priority date: 2020-07-29
Filing date: 2020-07-29
Publication date: 2025-01-03
Anticipated expiration: 2040-07-29
Also published as: CN114057724A

Abstract

The invention belongs to the field of pharmaceutical chemicals, and particularly relates to a BTK inhibitor, which aims to analyze the structural characteristics of the marketed medicine ibrutinib, combine the latest reported structure-activity relationship, ensure that the difference of N2-substituent R1 of piperidine-2, 6-dione has little influence on Btk enzyme activity, and the R1 faces one hydrophilic pocket of the enzyme, and introduce different hydrophilic groups to adjust the physicochemical properties of the compound. The N9 substituent R2 is spatially oriented towards the gatekeeper region of the kinase, and interacts with specific residues in the Tec family, and the difference exists in amino acid residues which interact with the gatekeeper region of different kinases, and the design and synthesis of higher cytotoxicity compounds are expected to exert anticancer effects and have pharmaceutical potential. Cytotoxicity experiments show that the compound has stronger in vitro inhibition effect on various cancer cells.

Description

BTK inhibitor

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a novel 3-amino-2, 7-dihydropyran [4,3-c ] pyrazole-4, 6-dione BTK inhibitor, and a preparation method and application thereof.

Background

Tumors are a common and frequently occurring disease that severely threatens human health, and are caused by abnormal proliferation of cells in the human body. Lung cancer, digestive tract tumor and liver cancer are the most common tumors in men and account 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 tumor and liver cancer are the most common tumors in women and account 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%). As the aging of the population of china increases, the prevalence of cancer tends to increase. In addition, environmental pollution increases the prevalence of some cancers, such as lymphoma. Autoimmune diseases are diseases in which the immune system reacts to the components of the body, causing damage.

BTK (brutontyrosinekinase) is a cytoplasmic protein belonging to the Tec family of non-receptor tyrosine kinases which are expressed in a large number of 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, with important effects on proliferation, differentiation and apoptosis of B cells.

In malignant B cells, the B cell receptor signaling pathway is overactive, thereby inhibiting normal differentiation and apoptosis of B cells and promoting abnormal proliferation. Abnormal regulation of BCR pathways is known to occur frequently in a variety of B cell type malignancies, such as Diffuse Large B Cell Lymphoma (DLBCL), mantle Cell Lymphoma (MCL), chronic Lymphocytic Leukemia (CLL), follicular Lymphoma (FL), giant globulinemia (WM), marginal Zone Lymphoma (MZL), B lymphoblastic leukemia (B-LBL), burkitt Lymphoma (BL), and the like.

Drug studies on BTK have not been stopped since the first discovery of BTK inhibitors in 1993. The indications for BTK inhibitors are still expanding, and besides anti-tumor, the use of autoimmune diseases will be a key to push BTK inhibitors to another market height. At present, a plurality of small molecule inhibitors of BTK are developed successively in China and have good anti-tumor effect in-vivo and in-vitro tests, more new BTK inhibitors are being studied except for drugs Ibrutinib and ACP-196 which are already on the market, and CT1530 and GS-4059 have entered clinical phase II except for BGB-3111 which is rapidly progressed at present

Currently, tumor targeted small molecule drugs aimed at BTK targets have been developed in generation 2, and the first generation BTK inhibitor is ibrutinib (Ibrutinib) marketed in 2013 in the united states for clinical treatment of recurrent or refractory Mantle Cell Lymphoma (MCL), which is a star drug targeting BTK, ibrutinib is capable of inhibiting the growth and metastasis of malignant proliferation B cells, while Ibrutinib is the first approved by the FDA for marketing. Ibrutinib is used in the treatment of autoimmune related diseases including stem cell transplantation, immune resistance after transplantation, arthritis, and the like, in addition to various hematological diseases. However Ibrutinib has not been a hundred percent of the response in treatment and the disease recurs, and there is unknown primary and acquired resistance to the drug. Along with the marketing and clinical use of drugs, ibrutinib began to develop resistance to different mechanisms in different patients. Acquired resistance only these two target mutations have been found so far, and mutation Ibrutinib occurs at 481 position of binding site of BTK and ibrutinib in BCR signal pathway, so that the problem of clinical resistance of disease Ibrutinib in arthritis inhibition model is more and more increased, and a plurality of side effects also occur.

The second generation BTK inhibitor Acalabrutinib (ACP-196), ACP-196 is referred to as a second generation BTK inhibitor because it does not inhibit other kinases on the basis of inhibiting BTK kinase targets. The FDA approved for the treatment of Mantle Cell Lymphoma (MCL) at month 8 of 2017 is currently in clinical use for many indications, e.g., chronic Lymphocytic Leukemia (CLL) is in clinical stage three, and some solid tumors such as non-small cell lung cancer, head and neck cancer, etc. are in clinical stage two and develop very rapidly. The treatment of arthritis with ACP-196 is also currently in clinical use due to the off-target effect of Ibrutinib. Thus, the BTK inhibitor with better selectivity in the second generation starts the period of onset of response, has little side effects due to its high selectivity, and has good development prospect in the treatment of chronic diseases such as autoimmune diseases.

The selectivity of the known BTK inhibitor is not ideal, and besides the BTK, other various kinases (such as ETK, EGF, BLK, FGR, HCK, YES, BRK, JAK3 and the like) are also inhibited, so that more side effects are generated, and meanwhile, drug resistance is often generated after mutation of a BTK binding site. Therefore, more BTK inhibitors are clinically needed for treating diseases such as tumors, and meanwhile, adverse events can be overcome.

The invention designs and synthesizes a novel 3-amino-2, 7-dihydropyran [4,3-c ] pyrazole-4, 6-diketone compound, and in vitro experiments show that the compound has better cytotoxicity to cancer cells, wherein the IC ₅₀ of the compound TM10 to lung cancer cells reaches 0.86 mu M, and the compound is a compound developed into a novel anticancer drug, and in the preparation method, the invention synthesizes a key intermediate and a target compound thereof efficiently by a simple method.

Disclosure of Invention

The invention aims to analyze the structural characteristics of the on-market medicine ibrutinib, combine the latest reported structure-activity relationship, have little influence on Btk enzyme activity by different N2-position substituents R1 of piperidine-2, 6-dione, and introduce different hydrophilic groups (such as acid, amide and amine) towards one hydrophilic pocket of the enzyme, so that the physicochemical properties of the compound can be regulated. The N9 substituent R2 is spatially oriented towards the gatekeeper region of the kinase and interacts with specific residues within the Tec family, differing in amino acid residues that interact with the gatekeeper region of different kinases:

It is desirable to design and synthesize more cytotoxic compounds that exert anticancer effects and possess pharmaceutical potential. Cytotoxicity experiments show that the compound has stronger in vitro inhibition effect on various cancer cells.

Interpretation of the terms

The following sets forth definitions of various terms used to describe the application. These definitions apply to the terms used throughout the specification and claims unless otherwise limited in specific instances, either alone or as part of a larger group.

The term "alkyl" in the present invention refers to saturated straight or branched chain hydrocarbon groups, in certain embodiments containing 1 to 4, respectively, examples of C _1-4 alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, or the like.

The term "aryl" as used herein refers to a mono-or polycyclic carbocyclic ring system having one or more aromatic rings, either fused or unfused, including but not limited to phenyl, naphthyl, tetrahydronaphthyl, and the like, and which ring carbon may also have hydrogen atoms substituted with one or more substituents.

The term "halo" as used herein refers to a group formed by substitution of a halogen atom for a hydrogen atom on a carbon atom, wherein the halogen atom includes, but is not limited to F, cl, br, I.

The term "alkoxy" in the present invention refers to-O-alkyl, wherein the alkyl includes, but is not limited to, C _1-3 alkyl and C _3-6 cycloalkyl, and specific examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, and halogenated versions thereof.

The term "alkylamino" as used herein refers to the group-N-alkyl, wherein the alkyl includes, but is not limited to, C _1-3 alkyl and C _3-6 cycloalkyl, and specific examples include, but are not limited to, methylamino, ethylamino, propylamino, cyclopropylamino, cyclobutoxy, and the like. The term "cycloalkyl" as used herein refers to a monovalent radical of a monocyclic or polycyclic saturated or partially unsaturated carbocyclic compound, and C _3-6 cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like, and the hydrogen atoms on the ring carbon may also be substituted with one or more substituents.

The term "heterocycloalkyl" according to the present invention refers to a mono-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, S.

The term "heteroaryl" according to the present invention refers to an aromatic ring system containing 1 to 6 carbons and at least one heteroatom selected from N, S, O.

The term "alkylamino" as used herein refers to the group-alkyl-NR _aR_b, wherein alkyl 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 are each independently selected from H, C _1-3 alkyl or C _3-6 cycloalkyl.

The term "pharmaceutically acceptable salt" as used herein refers to salts of the compounds of the present invention prepared from the compounds of the present invention having the specified substituents found herein and relatively non-toxic acids or bases. The relatively basic functional groups of the present invention may be present in pure solution or in a suitable inert solvent to obtain acid addition salts, including inorganic and organic acid salts, by contacting the neutral form of such compounds with a sufficient amount of the acid. The inorganic acid salts include, but are not limited to, hydrochloride, nitrate, borate, hydrocyanic acid salt, hydrofluoric acid salt, hydrobromide, hydroiodide, nitrite, perhalate, halate, hypohalite, meta-aluminate, sulfate, phosphate, nitrate, and the organic acid salts include, but are not limited to, formate, acetate, propionate, butyrate, acrylate, oxalate, malonate, succinate, benzoate, phthalate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, thioacetate, trifluoroacetate, tartrate, malate, citrate, ascorbate, salicylate, caffeic acid fumarate, maleate, lactate, citrate, glutamate, camphorsulfonate, and the like.

"Solvate" as used herein refers to a form of addition of a solvent, either stoichiometric or non-stoichiometric, selected from the group consisting of water, ethanol, isopropanol, diethyl ether, acetone, and the like.

"Prodrug" as used herein refers to a compound that is metabolically convertible in vivo to provide any compound described by the formulas of the present application, and various forms of drugs are known in the art.

An "effective amount" as used herein refers to a dosage that achieves the desired therapeutic effect in the desired subject without undue adverse effects, as may be generally determined by one of skill in the art as desired.

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

It should be understood that other terms not explained hereinabove but appearing in the present invention should be defined as commonly understood by one of ordinary skill in the art.

The specific technical scheme of the invention is as follows:

In a first aspect of the present invention, there is provided a hydrate, solvate, prodrug, stereoisomer or tautomer of a pyrazole-4, 6-dione compound of formula (I):

Wherein:

R is independently selected from one or more independent groups selected from H, halogen, aryl, heteroaryl, C _1-4 alkyl, C _1-6 alkoxy, C _3-6 cycloalkyl, C _2-6 heterocycloalkyl, aryloxy, -NR ₂R₃, or substituted aryl, substituted heteroaryl, substituted C _3-6 cycloalkyl, substituted C _2-6 heterocycloalkyl, substituted aryloxy, substituted C _1-6 alkoxy, wherein the substituent groups are selected from one or more substituents 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 heterocycloalkyl.

R is preferably-NR ₂R₃, aryl, heteroaryl, C _1-4 alkyl, C _2-6 heterocycloalkyl, substituted aryl, substituted heteroaryl.

R is further preferably-NR ₂R₃, aryl, heteroaryl, substituted aryl.

R ₂ is independently selected from H or C _1-4 alkyl, R ₂ is further preferably methyl.

R ₃ is independently selected from one or more of 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 heterocycloalkyl, and R ₃ is further preferably methyl.

Z is independently selected from C _1-4 alkyl or aryl, preferably methyl, ethyl, benzene, and more preferably methyl, benzene.

In a preferred embodiment of the invention, the compound of formula (I) is selected from the group consisting of:

In a second aspect, the application provides a method for preparing pyrazole-4, 6-dione compounds shown in formula (I). The compounds of the present application may be prepared using a variety of means known to those skilled in the art. The present application may be synthesized using the methods described herein, organic chemical synthesis methods, or variations thereof as understood by those skilled in the art. Preferred methods include, but are not limited to, those described below.

Preferably, the preparation method of the pyrazole-4, 6-dione compound shown in the formula (I) comprises the following steps:

preparation of Compound 6:

General synthetic procedure for compound 11:

Preparation of Compound I:

wherein R and Z are defined as the structural compounds of formula (I) of the first aspect of the invention;

Preferably, the preparation method of the pyrazole-4, 6-diketone compound shown in the formula (I) comprises the following reaction steps:

Step 1, performing condensation reaction on a compound 1 in a sodium ethoxide solution to obtain a compound 2;

step 2, reacting the compound 2 with hydrazine hydrate to obtain a compound 3;

step 3, reacting the compound 3 under an alkaline condition to obtain a compound 4;

Step 4, performing temperature-controlled reflux dehydration on the compound 4 to generate an intermediate compound 5;

and 5, reacting the compound 5 with 3-bromo-4-fluorobenzoic acid, EDCI and HOBt in dichloromethane to obtain a compound 6.

Step 6, the compound 8 and the compound 9 react to obtain a compound 10.

Step 7, hydrolyzing the compound 10 to obtain a compound 11.

And 8, reacting the compound 6 with the compound 11 in an organic solvent, crystallizing, filtering and drying to obtain the target compound I. Preferably, the preparation method of the pyrazole-4, 6-dione compound shown in the formula (I) comprises the following specific reaction steps:

And 1, adding sodium ethoxide into an ethanol solvent, stirring and dissolving, adding the compound 1, heating to reflux, detecting by TLC, controlling Wen Xijing after the reaction is finished, carrying out suction filtration, leaching a filter cake by using ethanol, drying to obtain a crude product compound 2, and recrystallizing the crude product to obtain a final compound 2.

Preferably, the crystallization temperature is 15-30 ℃.

And 2, adding the compound 2 into water, stirring and dissolving, adding hydrazine hydrate, heating to reflux, detecting by TLC, cooling and separating out solid after the reaction is finished, filtering and drying to obtain the compound 3.

Preferably, the crystallization temperature is-10-5 ℃.

And 3, adding the compound 3 into an alkali solution, stirring and dissolving, heating to reflux, detecting by TLC, cooling, adding hydrochloric acid to adjust the pH to 3-4 after the reaction is finished, precipitating a solid, filtering, and drying to obtain the compound 4.

Preferably, the base is sodium hydride, potassium hydroxide, sodium hydroxide, lithium hydroxide, and more preferably sodium hydroxide.

Step 4, dissolving the compound 4 in acetic anhydride, adding a condensing agent, heating to reflux, detecting by TLC, finishing the reaction,

Cooling, crystallizing, suction filtering, eluting filter cake with methyl tert-butyl ether, and drying to obtain compound 5.

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

And 5, dissolving 3-bromo-4-fluorobenzoic acid in dried dichloromethane, sequentially adding EDCI, HOBt and a compound 5, stirring at room temperature for reaction, detecting by TLC, filtering, concentrating, evaporating under reduced pressure, and separating by a column layer to obtain the compound 6.

And 6, adding the compound 8 into dried dichloromethane, sequentially adding the compound 9 and a condensing agent, stirring at room temperature, detecting by TLC, and evaporating under reduced pressure to dryness after the reaction is finished, and separating by a column layer to obtain the compound 10.

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

And 7, dissolving the compound 10 in an organic solvent, dropwise adding hydrochloric acid with the concentration of 3mol/L, removing Boc groups, washing with purified water, drying an organic layer by using anhydrous sodium sulfate, filtering, concentrating, and evaporating under reduced pressure to obtain a compound 11.

Preferably, the organic solvent is ethyl acetate, dichloromethane, methanol, ethanol, wherein ethyl acetate is preferred.

And 8, dissolving the compound 6 in an organic solvent, adding the compound 11, heating to reflux reaction, detecting the end of the reaction by TLC, cooling and crystallizing, leaching a filter cake by the organic solvent, and drying to obtain the compound I.

Preferably, the organic solvent is toluene, benzene, xylene, and toluene is preferred.

Preferably, the crystallization temperature is-5 ℃, wherein 0 ℃ is preferred.

In a third aspect of the present invention there is provided a pharmaceutical composition comprising a therapeutically effective amount of one or more of the compounds of formula (I) of the present invention or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer or tautomer thereof, and optionally a pharmaceutically acceptable carrier, excipient, adjuvant or diluent.

Preferably, the pharmaceutical composition is useful for treating diseases associated with abnormal expression of tyrosine kinase or higher activity of tyrosine kinase.

Preferably, the diseases associated with abnormal expression of tyrosine kinase or high activity of tyrosine kinase include, but are not limited to, abnormal proliferation of cells, morphological changes, hyperkinesia, angiogenesis diseases, tumor growth, and tumor metastasis diseases.

Preferably, a BTK inhibitor comprises an inhibiting effective amount of one or more of pyrazole-4, 6-dione compounds of formula (I), or a pharmaceutically acceptable salt, tautomer, optical isomer, pharmaceutically acceptable solvate thereof, according to the invention, and optionally a pharmaceutically acceptable carrier, excipient, adjuvant or diluent.

The compounds of the invention may be used directly for prophylaxis and therapy or preferably in the form of pharmaceutical compositions. Although the active ingredients can be administered alone, it is preferably in the form of a pharmaceutical preparation or composition. Accordingly, the present invention provides a pharmaceutical formulation comprising a compound of the present invention together with a pharmaceutically acceptable diluent, excipient or carrier (collectively referred to herein as "carrier" materials). The pharmaceutical composition of the present invention may take the form of a pharmaceutical formulation as described below. The present invention therefore relates to a pharmaceutical composition comprising at least one compound of formula (I) and conventional excipients.

Exemplary compositions for oral administration include suspending agents which may comprise, for example, microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancing agent, and sweeteners or flavoring agents such as are known in the art, such as immediate release tablets which may comprise, for example, microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate, calcium sulfate, sorbitol, glucose and/or lactose and/or other excipients, binders, bulking agents, disintegrants, diluents and lubricants such as are known in the art. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, and the like. The compounds of formula (I) may also be delivered orally by sublingual and/or buccal means. Molded, compressed or lyophilized tablets are exemplary forms that may be used. Exemplary compositions include those formulated with the compounds of the present invention with fast dissolving diluents such as mannitol, lactose, sucrose, and/or cyclodextrins. High molecular weight excipients such as cellulose (microcrystalline powdered cellulose) or polyethylene glycol (PEG) may also be included in these formulations. These formulations may also include excipients that aid in mucoadhesion, such as hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), sodium carboxymethyl cellulose (SCMC), maleic anhydride copolymers (e.g., gantrez) and agents that control release, such as polyacrylic copolymers (e.g., carbopo 1934). Lubricants, glidants, flavoring agents, coloring agents and stabilizers may also be added to facilitate preparation and use. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. For oral administration in liquid form, the oral pharmaceutical component may be combined with any orally acceptable, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.

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

Formulations of the invention suitable for oral administration may be in the form of various discrete 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 or water-in-oil emulsions. The active ingredient may also be formulated as a bolus, electuary or paste.

Tablets may be made by compression or molding with the optional auxiliary ingredient(s). Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as powder or granules optionally mixed with a binder, lubricant, inert diluent, lubricant, surfactant or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein. The compounds of the present invention may be administered in a form suitable for immediate or sustained release. Immediate or sustained release may be achieved by the use of a suitable pharmaceutical composition comprising a compound of the invention, or, especially in the case of sustained release, by means such as subcutaneous implants or osmotic pumps. The compounds of the invention may also be administered in a liposomal manner. Preferred unit dosage formulations are those containing an effective dose (as hereinafter described) or suitable portion thereof of the active ingredient.

It will be appreciated that in addition to the ingredients specifically mentioned above, the formulations of the present invention may also include other agents conventional in the art relating to the type of formulation under investigation, for example, formulations suitable for oral administration may include flavouring agents.

The formulations may be conveniently presented as unit dosage forms and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients. Typically, the active ingredient is homogeneously and intimately associated with a liquid carrier and/or a finely divided solid carrier to manufacture a formulation, which product is then, if necessary, shaped to give the desired formulation.

The compounds of the invention may 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 a variety of phospholipids, 1, 2-dipalmitoyl phosphatidylcholine, phosphatidylethanolamine (cephalin), phosphatidylserine, phosphatidylinositol, dipeptidyl glycerol (cardiolipin), or phosphatidylcholine (lecithin).

Formulations for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions which may contain suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, saline or water-for-injection immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described. Exemplary compositions for parenteral administration include injectable solutions or suspensions which may contain, for example, suitable non-toxic parenterally acceptable diluents or solvents, such as polyethylene glycol, ethanol, 1, 3-butanediol, water, ringer's solution, sodium chloride isotonic solution, or other suitable dispersing or wetting agents and suspending agents, including synthetic mono-or diglycerides and fatty acids, including oleic acid and Cremaphor.

Exemplary compositions for nasal, aerosol, or inhalation 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.

Formulations for rectal administration may be presented as suppositories using conventional carriers such as cocoa butter/synthetic glycerides or polyethylene glycols. These carriers are typically solid at ambient conditions, but liquefy and/or dissolve within the rectal lumen to release the drug.

Formulations for topical administration in the mouth, e.g. buccal or sublingual administration, include lozenges comprising the active ingredient in a flavoured base such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a base of gelatin and glycerol or sucrose and acacia. Exemplary compositions for topical administration include topical carriers such as Plastibase (mineral oil rubberized with polyethylene).

In a fourth aspect, the present invention provides the use of pyrazole-4, 6-dione compounds of formula (I), in particular for the preparation of tyrosine kinase inhibitors, for the in vitro non-therapeutic inhibition of tyrosine kinase activity, for the in vitro non-therapeutic inhibition of tumour cell growth or a combination thereof, for the treatment or prophylaxis of diseases in which BTK plays a role.

Preferably, the pyrazole-4, 6-dione compound shown in the formula (I) is used for preparing a medicament for treating or preventing diseases in which BTK plays a role.

Preferably, the disease in which BTK plays a role includes cancer or autoimmune system-related disease, particularly including but not limited to non-small cancer lung cells, lymphomas, breast cancer, leukemia and head and neck squamous cell 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's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal stromal tumor, thyroid cancer, cholangiocarcinoma, endometrial cancer, renal cancer, anaplastic large cell lymphoma, acute myelogenous 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, syndrome of pulmonary hemorrhagic nephritis, primary biliary cirrhosis, multiple cerebral spinal sclerosis, acute idiopathic polyneuritis, systemic lupus erythematosus, rheumatoid arthritis, systemic vasculitis, scleroderma, tenascus, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia, thyroid autoimmune disease, ulcerative colitis.

Preferably, the treatment is a method of alleviating or alleviating the symptoms of the disease or condition and complications thereof, and the prevention is the reduction or elimination of the symptoms or complications of the disease, condition or disorder.

Compared with the prior art, the invention has the main advantages that:

(1) The invention provides a pyrazole-4, 6-dione compound with a novel structure.

(2) The pyrazole-4, 6-dione compound of the formula (I) provided by the invention has remarkable inhibition effect on the growth of tumor cells.

(3) The pyrazole-4, 6-dione compound of the formula (I) provided by the invention has the advantages of cheap and easily available starting materials, shorter synthetic route, simpler operation and suitability for industrial scale-up production.

It is to be understood that within the scope of the present invention, the above-described technical features of the present invention and technical features specifically described below (including embodiments) may be combined with each other to constitute new or preferred technical solutions.

Detailed Description

The process according to the invention is further illustrated by the following examples. It should be understood that the preparation method of the embodiment of the present invention is merely for illustrating the present invention, and not for limiting the present invention, and simple modification of the preparation method of the present invention under the premise of the concept of the present invention falls within the scope of the present invention as claimed.

Abbreviations used herein:

DCC dicyclohexylcarbodiimide

EDCI 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

DMC 2-chloro-1, 3-dimethylimidazoline chloride

HOBt 1-carbonyl benzotriazole

TLC thin layer chromatography

Boc group t-Butoxycarbonyl group

Preparation of Compound 6:

Adding sodium ethoxide solid (4 g,0.01 mol) into absolute ethyl alcohol (40 ml), stirring at room temperature for dissolution, pouring (13.20 g,0.02 mol) malononitrile, slowly heating to reflux, detecting by TLC, controlling the temperature to 20 ℃ after the reaction is finished, stirring for crystallization, completing crystallization, leaching by ethanol, transferring the obtained solid to a three-mouth bottle after decompression and drying, pouring purified water, stirring at room temperature for dissolution, adjusting the pH value to 3-4 by using 5mol/L hydrochloric acid, precipitating crystals, filtering, recrystallizing a filter cake by using purified water, obtaining the compound 2, drying under reduced pressure, wherein the yield is 85.6%, and the purity is 99.92%.

Preparation of compound 3:

Adding the intermediate 2 into 500ml of water, stirring for dissolution, adding 50ml of hydrazine hydrate, slowly heating to reflux, detecting by TLC, cooling to 0 ℃ for crystallization after the reaction is finished, filtering to precipitate, and drying under reduced pressure to obtain the compound 3, wherein the yield is 87.8% and the purity is 99.89%.

Preparation of Compound 4:

and adding the compound 3 into 0.2M NaOH (200 ml), stirring and dissolving, heating to reflux reaction, detecting by TLC, cooling to 0 ℃ after the reaction is finished, regulating the pH of the reaction solution to 3-4 by using 5mol/L hydrochloric acid, filtering, and drying under reduced pressure to obtain an intermediate 4, wherein the yield is 92.5%, and the purity is 99.90%.

Preparation of compound 5:

Compound 4 (150 g,0.90 mol) was dissolved in acetic anhydride (500 ml), DCC (240.5 g,1.17 mol) was added, heated to reflux, checked by TLC, cooled to 0 ℃ after the reaction was completed, crystallized, suction filtered, the filter cake rinsed with methyl tert-butyl ether, dried under reduced pressure to give compound 5 in 91.7% yield with 99.88% purity.

Preparation of Compound 6:

3-bromo 4-fluorobenzoic acid (2.19 g,0.1 mol) was added to 300ml of methylene chloride, 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 this order, and the reaction was stirred at room temperature, and then subjected to TLC detection, filtration, concentration, evaporation under reduced pressure and separation by column layer to give Compound 6 in a yield of 90.5% and a purity of 99.87%.

Example 1

Preparation of compound TM 1:

Preparation of Compound 10-1:

Compound 8 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-1 (8.90 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, and the reaction was completed by TLC, evaporated to dryness under reduced pressure, and separated by column layer to give compound 10-1 in a yield of 91.8% and a purity of 99.78%.

Preparation of Compound 11-1:

Compound 10-1 (23.13 g,0.1 mol) was dissolved in 150mL of ethyl acetate, 30mL of hydrochloric acid having a concentration of 3mol/L was added dropwise, the Boc group was removed, 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 give Compound 11 in a yield of 90.3% and a purity of 99.85%.

Preparation of compound TM 1:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-1 (15.74 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM1 in a yield of 92.4% and a purity of 99.92%.

¹HNMR(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);

¹³CNMR(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.

MS(m/z):481.24[M+H]⁺;

Example 2

Preparation of compound TM 2:

preparation of Compound 10-2:

Compound 8 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-2 (12.89 g,0.13 mol), HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, and TLC was checked, and after completion of the reaction, evaporated to dryness under reduced pressure, compound 10-2 was isolated by column layer in 91.5% yield and 99.77% purity.

Preparation of Compound 11-2:

Compound 10-2 (25.64 g,0.1 mol) was dissolved in 150mL of ethyl acetate, 30mL of hydrochloric acid having a concentration of 3mol/L was added dropwise, the Boc group was removed, 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 give compound 11-2 in a yield of 91.1% and a purity of 99.89%.

Preparation of compound TM 2:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-2 (18.75 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM2 in a yield of 92.1% and a purity of 99.90%.

¹HNMR(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);

¹³C-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).

MS(m/z):506.12[M+H]⁺;

Example 3

Preparation of compound TM 3:

Preparation of Compound 10-3:

compound 8 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-3 (11.18 g,0.12 mol), HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, and TLC was checked, and after completion of the reaction, evaporated to dryness under reduced pressure, compound 10-3 was isolated by column layer in 92.0% yield and 99.82% purity.

Preparation of Compound 11-3:

Compound 10-3 (25.03 g,0.1 mol) was dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid having a concentration of 3mol/L was added dropwise, the Boc group was removed, 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 give compound 11-3 in a yield of 91.6% and a purity of 99.90%.

Preparation of compound TM 3:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-3 (18.02 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM3 in a yield of 92.1% and a purity of 99.90%.

¹HNMR(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);

¹³C-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.

MS(m/z):500.18[M+H]⁺;

Example 4

Preparation of Compound TM 4:

preparation of Compound 10-4:

Compound 8 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-4 (13.33 g,0.12 mol), HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, and TLC was checked, and after completion of the reaction, evaporated to dryness under reduced pressure, compound 10-4 was isolated by column layer in 92.2% yield and 99.85% purity.

Preparation of Compound 11-4:

Compound 10-4 (26.83 g,0.1 mol) was dissolved in 150mL of ethyl acetate, 30mL of hydrochloric acid with a concentration of 3mol/L was added dropwise, boc groups were removed, 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 give compound 11-4 in a yield of 91.8% and a purity of 99.91%.

Preparation of Compound TM 4:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-4 (20.18 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM4 in a yield of 91.7% and a purity of 99.87%.

¹HNMR(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);

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(2C),127.5,121.8(2C),118.7,112.2,98.7,54.8,30.5.

MS(m/z):518.31[M+H]⁺;

Example 5

Preparation of compound TM 5:

preparation of Compound 10-5:

compound 8 (18.92 g,0.1 mol) was added to dried dichloromethane, and compound 9-5 (11.29 g,0.12 mol), HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, and the reaction was completed by TLC, evaporated to dryness under reduced pressure, and separated by column layer to give compound 10-5 in a yield of 91.9% and a purity of 99.81%.

Preparation of Compound 11-5:

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

Preparation of compound TM 5:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-5 (18.14 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM5 in a yield of 91.9% and a purity of 99.89%.

¹HNMR(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);

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.

MS(m/z):501.16[M+H]⁺;

Example 6

Preparation of compound TM 6:

Preparation of Compound 10-6:

Compound 8 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-6 (16.46 g,0.12 mol), HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, and TLC was checked, and after completion of the reaction, evaporated to dryness under reduced pressure, compound 10-6 was isolated by column layer in 92.2% yield with 99.84% purity.

Preparation of Compound 11-6:

compound 10-6 (29.44 g,0.1 mol) was dissolved in 150mL of ethyl acetate, 30mL of hydrochloric acid having a concentration of 3mol/L was added dropwise, the Boc group was removed, 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 give compound 11-6 in a yield of 92.2% and a purity of 99.91%.

Preparation of compound TM 6:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-6 (23.71 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM6 in a yield of 92.4% and a purity of 99.91%.

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);

¹³C-NMR(100MHz,DMSO-d₆):168.4,167.9,157.6,155.5,150.2,141.1,135.1,133.4,132.5,131.0,129.5,128.1,125.4,123.3,122.8,120.9,118.7,112.2,98.7,56.4,54.8,27.0,18.2.

MS(m/z):544.22[M+H]⁺;

Example 7

Preparation of compound TM 7:

Preparation of Compound 10-7:

Compound 8 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-7 (15.98 g,0.12 mol), HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, and TLC was checked, and after completion of the reaction, evaporated to dryness under reduced pressure, compound 10-7 was isolated by column layer in 92.0% yield and 99.82% purity.

Preparation of Compounds 11-7:

Compound 10-7 (29.03 g,0.1 mol) was dissolved in 150mL of ethyl acetate, 30mL of hydrochloric acid having a concentration of 3mol/L was added dropwise, the Boc group was removed, 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 give compound 11-7 in a yield of 91.8% and a purity of 99.88%.

Preparation of compound TM 7:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-7 (22.82 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM7 in a yield of 92.0% and a purity of 99.87%.

¹HNMR(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);

¹³C-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.

MS(m/z):540.13[M+H]⁺;

Example 8

Preparation of Compound TM 8:

Preparation of Compound 10-8:

Compound 8 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-8 (32.93 g,0.12 mol), HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, and TLC was checked, and after completion of the reaction, evaporated to dryness under reduced pressure, compound 10-8 was isolated by column layer, yield 92.5%, purity 99.87%.

Preparation of Compounds 11-8:

Compound 10-8 (43.16 g,0.1 mol) was dissolved in 150mL of ethyl acetate, 30mL of hydrochloric acid having a concentration of 3mol/L was added dropwise, boc groups were removed, 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 give compound 11-8 in a yield of 92.4% and a purity of 99.90%.

Preparation of Compound TM 8:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-8 (39.78 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM8 in a yield of 92.3% and a purity of 99.90%.

¹HNMR(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);

¹³C-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),118.7,118.4,118.3,117.7(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.

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

Example 9

Preparation of Compound TM 9:

Preparation of Compounds 10-9:

Compound 8 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-9 (16.46 g,0.12 mol), HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, and TLC was checked, and after completion of the reaction, evaporated to dryness under reduced pressure, compound 10-9 was isolated by column layer in 92.1% yield and 99.85% purity.

Preparation of Compounds 11-8:

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

Preparation of Compound TM 9:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-9 (23.31 g,0.12 mol) was added, the mixture was heated to reflux, TLC detection was completed, the temperature was lowered to 0℃for crystallization, crystallization was completed, filtration was carried out, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to obtain compound TM9 in a yield of 92.0% and a purity of 99.89%.

MS(m/z):544.15[M+H]⁺;¹H-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);¹³C-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.

Example 10

Preparation of compound TM 10:

preparation of Compounds 10-10:

Compound 8-1 (23.73 g,0.1 mol) was added to dried dichloromethane, compound 9-10 (16.46 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-10 was isolated by column layer, yield 91.7%, purity 99.82%.

Preparation of Compounds 11-10:

Compound 10-10 (35.64 g,0.1 mol) is dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with the concentration of 3mol/L is added dropwise, boc groups are removed, purified water is used for washing, an organic layer is dried by anhydrous sodium sulfate, filtration, concentration and reduced pressure evaporation are carried out, and compound 11-10 is obtained, the yield is 92.0%, and the purity is 99.87%.

Preparation of compound TM 10:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-10 (30.76 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM10 in a yield of 92.2% and a purity of 99.91%.

¹H-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);

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.

MS(m/z):606.20[M+H]⁺;

Example 11

Preparation of Compound TM 11:

preparation of Compounds 10-11:

Compound 8-1 (23.73 g,0.1 mol) was added to dried dichloromethane, compound 9-11 (8.90 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-11 was isolated by column layer, yield 92.0%, purity 99.85%.

Preparation of Compounds 11-11:

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

Preparation of Compound TM 11:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-11 (23.19 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM11 in a yield of 92.0% and a purity of 99.88%.

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);

¹³C-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.

MS(m/z):543.13[M+H]⁺;

Example 12

Preparation of Compound TM 12:

preparation of Compounds 10-12:

Compound 8-1 (23.73 g,0.1 mol) was added to dried dichloromethane, compound 9-12 (8.90 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-12 was isolated by column layer, yield 92.0%, purity 99.85%.

Preparation of Compounds 11-12:

Compound 10-12 (29.34 g,0.1 mol) is dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with the concentration of 3mol/L is added dropwise, boc groups are removed, purified water is used for washing, an organic layer is dried by anhydrous sodium sulfate, filtration, concentration and reduced pressure evaporation are carried out, and compound 11-12 is obtained, the yield is 92.4%, and the purity is 99.89%.

Preparation of Compound TM 12:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-12 (23.19 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM12 in a yield of 92.0% and a purity of 99.88%.

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),3.59(s,2H),3.49～3.41(m,1H),1.59～1.22(m,10H);

¹³C-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.

MS(m/z):568.09[M+H]⁺;

Example 13

Preparation of Compound TM 13:

Preparation of Compounds 10-13:

Compound 8-1 (23.73 g,0.1 mol) was added to dried dichloromethane, compound 9-13 (11.18 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-13 was isolated by column layer, yield 91.8%, purity 99.84%.

Preparation of Compounds 11-13:

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

Preparation of Compound TM 13:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-13 (25.47 g,0.12 mol) was added, the mixture was heated to reflux, the reaction was checked by TLC, cooled to 0℃for crystallization, the crystallization was completed, filtered, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM13 in a yield of 91.7% and a purity of 99.82%.

¹H-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);

¹³C-NMR(400MHz,DMSO-d₆):δ168.8,167.7,162.8,158.6,149.7,142.4,137.5,136.7,131.2,130.4,129.1(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.

MS(m/z):562.21[M+H]⁺;

Example 14

Preparation of Compound TM 14:

preparation of Compounds 10-14:

Compound 8-1 (23.73 g,0.1 mol) was added to dried dichloromethane, compound 9-14 (13.33 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-14 was isolated by column layer, yield 91.4%, purity 99.81%.

Preparation of Compounds 11-14:

Compound 10-14 (33.04 g,0.1 mol) was dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with a concentration of 3mol/L was added dropwise, boc groups were removed, the mixture was washed with purified water, and the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, evaporated to dryness under reduced pressure to give compound 11-14 in a yield of 92.0% and a purity of 99.87%.

Preparation of Compound TM 14:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-14 (27.63 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM14 in a yield of 91.1% and a purity of 99.80%.

¹H-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);

¹³C-NMR(400MHz,DMSO-d₆):δ168.8,167.7,162.8,161.5,158.6,149.7,142.4,137.5,136.7,133.4,130.4,128.5(2C),125.8(2C),127.7(2C),123.4,122.5,121.2,119.5,117.1(2C),110.5,99.5,31.2.

MS(m/z):580.15[M+H]⁺;

Example 15

Preparation of Compound TM 15:

Preparation of Compounds 10-15:

compound 8-1 (23.73 g,0.1 mol) was added to dried dichloromethane, compound 9-15 (11.29 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-15 was isolated by column layer, yield 92.0%, purity 99.87%.

Preparation of Compounds 11-15:

Compound 10-15 (31.34 g,0.1 mol) is dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with the concentration of 3mol/L is added dropwise, boc groups are removed, purified water is used for washing, an organic layer is dried by anhydrous sodium sulfate, filtration, concentration and reduced pressure evaporation are carried out, and compound 11-15 is obtained, the yield is 92.3%, and the purity is 99.89%.

Preparation of Compound TM 15:

intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-15 (25.59 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM15 in a yield of 91.9% and a purity of 99.88%.

¹H-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);

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.

MS(m/z):563.11[M+H]⁺;

Example 16

Preparation of Compound TM 16:

preparation of Compounds 10-16:

Compound 8-1 (23.73 g,0.1 mol) was added to dried dichloromethane, compound 9-16 (17.18 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-16 was isolated by column layer, yield 91.5%, purity 99.84%.

Preparation of Compounds 11-16:

Compound 10-16 (36.24 g,0.1 mol) was dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with a concentration of 3mol/L was added dropwise, boc groups were removed, 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 give compound 11-16 in a yield of 91.8% and a purity of 99.82%.

Preparation of Compound TM 16:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-16 (31.48 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM16 in a yield of 91.4% and a purity of 99.85%.

¹H-NMR(400MHz,DMSO-d₆):δ11.03(s,1H),10.20(s,1H),8.74(s,1H),8.03～7.92(m,2H),7.85～7.71(m,4H),7.57～7.41(m,4H),7.25～7.17(m,4H),3.59(s,2H);

¹³C-NMR(400MHz,DMSO-d₆):δ168.8,167.7,162.8,158.6,154.3,149.7,146.8,142.4,139.4,137.5,135.7,133.4,132.8,130.4,129.1,128.5(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.

MS(m/z):612.16[M+H]⁺;

Example 17

Preparation of Compound TM 17:

preparation of Compounds 10-17:

Compound 8-1 (23.73 g,0.1 mol) was added to dried dichloromethane, compound 9-17 (15.98 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-17 was isolated by column layer, yield 91.9%, purity 99.86%.

Preparation of Compounds 11-17:

Compound 10-17 (33.52 g,0.1 mol) is dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with the concentration of 3mol/L is added dropwise, boc groups are removed, purified water is used for washing, an organic layer is dried by anhydrous sodium sulfate, filtration, concentration and reduced pressure evaporation are carried out, and compound 11-17 is obtained, the yield is 92.1%, and the purity is 99.89%.

Preparation of Compound TM 17:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-17 (30.27 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM17 in a yield of 91.7% and a purity of 99.86%.

¹H-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);

¹³C-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;

MS(m/z):602.18[M+H]⁺;

Example 18

Preparation of Compound TM 18:

preparation of Compounds 10-18:

Compound 8-1 (23.73 g,0.1 mol) was added to dried dichloromethane, compound 9-18 (32.93 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-18 was isolated by column layer, yield 92.4%, purity 99.88%.

Preparation of Compounds 11-18:

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

Preparation of Compound TM 18:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compounds 11-18 (47.22 g,0.12 mol) were added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM18 in a yield of 92.1% and a purity of 99.88%.

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),7.31～7.22(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);

¹³C-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).

MS(m/z):743.24[M+H]⁺;

Example 19

Preparation of Compound TM 19:

preparation of Compounds 10-19:

Compound 8-1 (23.73 g,0.1 mol) was added to dried dichloromethane, compound 9-19 (17.79 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-19 was isolated by column layer, yield 91.7%, purity 99.80%.

Preparation of Compounds 11-19:

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

Preparation of Compound TM 19:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compounds 11-19 (32.20 g,0.12 mol) were added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM19 in a yield of 91.6% and a purity of 99.84%.

¹H-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);

¹³C-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.

MS(m/z):618.19[M+H]⁺;

Example 20

Preparation of Compound TM 20:

Preparation of Compounds 10-20:

Compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-20 (8.90 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-20 was isolated by column layer, yield 91.1%, purity 99.75%.

Preparation of Compounds 11-20:

Compound 10-20 (24.53 g,0.1 mol) is dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with the concentration of 3mol/L is added dropwise, boc groups are removed, purified water is used for washing, an organic layer is dried by anhydrous sodium sulfate, filtration, concentration and reduced pressure evaporation are carried out, and compound 11-20 is obtained, the yield is 91.2%, and the purity is 99.83%.

Preparation of Compound TM 20:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-20 (17.43 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM20 in a yield of 91.0% and a purity of 99.80%.

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),4.88～4.81(m,2H),4.31(s,2H),3.59(s,2H),2.71～2.64(m,2H),2.26(s,6H);

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.

MS(m/z):495.32[M+H]⁺;

Example 21

Preparation of Compound TM 21:

preparation of Compounds 10-21:

compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-21 (11.90 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-21 was isolated by column layer, yield 91.4%, purity 99.78%.

Preparation of Compounds 11-21:

Compound 10-21 (27.04 g,0.1 mol) was dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid having a concentration of 3mol/L was added dropwise, the Boc group was removed, 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 give compound 11-21 in a yield of 91.9% and a purity of 99.88%.

Preparation of Compound TM 21:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-21 (20.43 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM21 in a yield of 92.1% and a purity of 99.89%.

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);

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

MS(m/z):520.11[M+H]⁺;

Example 22

Preparation of compound TM 22:

preparation of Compounds 10-22:

Compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-22 (11.18 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-22 was isolated by column layer, yield 91.6%, purity 99.79%.

Preparation of Compounds 11-22:

Compound 10-22 (26.43 g,0.1 mol) is dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with the concentration of 3mol/L is added dropwise, boc groups are removed, purified water is used for washing, an organic layer is dried by anhydrous sodium sulfate, filtration, concentration and reduced pressure evaporation are carried out, and compound 11-22 is obtained, the yield is 91.1%, and the purity is 99.82%.

Preparation of compound TM 22:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-22 (19.71 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM22 in a yield of 91.5% and a purity of 99.82%.

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,6H),4.31(s,2H),3.59(s,2H),2.71～2.64(m,2H);

¹³C-NMR(400MHz,DMSO-d₆):δ170.1,168.6,167.8,159.2,149.7,145.6,142.4,139.5,133.1,132.3,128.8(2C),128.2,127.1,121.8(2C),118.5,111.0,99.6,40.1,33.7,31.2.

MS(m/z):514.17[M+H]⁺;

Example 23

Preparation of compound TM 23:

preparation of Compounds 10-23:

Compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-23 (13.33 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-23 was isolated by column layer, yield 91.0%, purity 99.74%.

Preparation of Compounds 11-23:

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

Preparation of compound TM 23:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compounds 11-23 (21.86 g,0.12 mol) were added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM23 in a yield of 91.7% and a purity of 99.84%.

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,5H),4.31(s,2H),3.59(s,2H),2.71～2.64(m,2H);

¹³C-NMR(400MHz,DMSO-d₆):δ170.1,168.6,167.8,162.3,159.2,149.7,145.6,142.4,133.1,132.3,131.5,128.2,121.8(2C),118.5,115.2(2C),111.0,99.6,40.1,33.7,31.2.

MS(m/z):532.24[M+H]⁺;

Example 24

Preparation of Compound TM 24:

preparation of Compounds 10-24:

Compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-24 (11.29 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-24 was isolated by column layer, yield 91.6%, purity 99.77%.

Preparation of Compounds 11-24:

Compound 10-24 (26.53 g,0.1 mol) is dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with the concentration of 3mol/L is added dropwise, boc groups are removed, purified water is used for washing, an organic layer is dried by anhydrous sodium sulfate, filtration, concentration and reduced pressure evaporation are carried out, and compound 11-24 is obtained, the yield is 91.8%, and the purity is 99.85%.

Preparation of Compound TM 24:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-24 (19.82 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM24 in a yield of 91.8% and a purity of 99.87%.

¹H-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);

¹³C-NMR(400MHz,DMSO-d₆):δ170.1,168.6,167.8,159.2,151.3,149.7,147.3,145.6,142.4,139.2,133.1,132.3,128.2,124.5,118.5,115.6,111.0,99.6,40.1,33.7,31.2.

MS(m/z):515.19[M+H]⁺;

Example 25

Preparation of Compound TM 25:

preparation of Compounds 10-25:

Compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-25 (16.46 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-25 was isolated by column layer, yield 91.0%, purity 99.71%.

Preparation of Compounds 11-25:

Compound 10-25 (30.84 g,0.1 mol) is dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with the concentration of 3mol/L is added dropwise, boc groups are removed, purified water is used for washing, an organic layer is dried by anhydrous sodium sulfate, filtration, concentration and reduced pressure evaporation are carried out, and compound 11-25 is obtained, the yield is 91.3%, and the purity is 99.83%.

Preparation of Compound TM 25:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-25 (24.99 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM25 in a yield of 91.0% and a purity of 99.81%.

¹H-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);

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.

MS(m/z):558.07[M+H]⁺;

Example 26

Preparation of Compound TM 26:

Preparation of Compounds 10-26:

Compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-26 (15.98 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-26 was isolated by column layer, yield 91.9%, purity 99.82%.

Preparation of Compounds 11-26:

Compound 10-26 (30.44 g,0.1 mol) was dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with a concentration of 3mol/L was added dropwise, boc groups were removed, 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 give compound 11-26 in a yield of 91.9% and a purity of 99.87%.

Preparation of Compound TM 26:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compounds 11-26 (24.51 g,0.12 mol) were added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM26 in a yield of 92.2% and a purity of 99.90%.

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);

¹³C-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.

MS(m/z):554.30[M+H]⁺;

Example 27

Preparation of compound TM 27:

preparation of Compounds 10-27:

Compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-27 (32.93 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-27 was isolated by column layer, yield 92.3%, purity 99.87%.

Preparation of Compounds 11-27:

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

Preparation of compound TM 27:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-27 (41.46 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM27 in a yield of 92.4% and a purity of 99.91%.

¹H-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);

¹³C-NMR(400MHz,DMSO-d₆):δ170.1,168.6,167.8,159.2,149.7,146.6,145.6,142.4,133.1,132.3,128.2,127.3,126.1(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).

MS(m/z):695.27[M+H]⁺;

Example 28

Preparation of Compound TM 28:

preparation of Compounds 10-28:

Compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-28 (18.02 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-28 was isolated by column layer, yield 91.5%, purity 99.82%.

Preparation of Compounds 11-28:

Compound 10-28 (32.14 g,0.1 mol) is dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with the concentration of 3mol/L is added dropwise, boc groups are removed, purified water is used for washing, an organic layer is dried by anhydrous sodium sulfate, filtration, concentration and reduced pressure evaporation are carried out, and compound 11-28 is obtained, the yield is 91.1%, and the purity is 99.80%.

Preparation of Compound TM 28:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compounds 11-28 (26.55 g,0.12 mol) were added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM28 in a yield of 91.8% and a purity of 99.87%.

¹H-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);

¹³C-NMR(400MHz,DMSO-d₆):δ174.0,170.1,168.6,167.8,159.2,155.2,149.7,145.6,142.4,133.1,132.3,131.2,128.2,122.5,123.6,120.3,119.1,118.5,111.0,99.6,40.1,33.7,31.2.

MS(m/z):571.22[M+H]⁺;

Example 29

Preparation of compound TM 29:

Preparation of Compounds 10-29:

Compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-29 (17.18 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-29 was isolated by column layer, yield 91.2%, purity 99.81%.

Preparation of Compounds 11-29:

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

Preparation of compound TM 29:

Intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compounds 11-29 (25.71 g,0.12 mol) were added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM29 in a yield of 91.4% and a purity of 99.85%.

¹H-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);

¹³C-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.

MS(m/z):564.16[M+H]⁺;

Example 30

Preparation of Compound TM 30:

preparation of Compounds 10-30:

Compound 8-2 (18.92 g,0.1 mol) was added to dried dichloromethane, compound 9-30 (15.31 g,0.12 mol) and HOBt (13.51 g,0.1 mol), EDCI (19.17 g,0.1 mol) were added in this order, stirred at room temperature, TLC was checked, the reaction was completed, evaporated to dryness under reduced pressure, and compound 10-30 was isolated by column layer, yield 91.6%, purity 99.83%.

Preparation of Compounds 11-30:

Compound 10-30 (29.88 g,0.1 mol) is dissolved in 180mL of ethyl acetate, 30mL of hydrochloric acid with the concentration of 3mol/L is added dropwise, boc groups are removed, purified water is used for washing, an organic layer is dried by anhydrous sodium sulfate, filtration, concentration and reduced pressure evaporation are carried out, and compound 11-30 is obtained, the yield is 91.9%, and the purity is 99.88%.

Preparation of Compound TM 30:

intermediate 6 (18.41 g,0.05 mol) was added to a 100ml three-necked flask, 80ml toluene was poured, compound 11-30 (23.84 g,0.12 mol) was added, the reaction was heated to reflux, the reaction was checked by TLC, the temperature was lowered to 0℃for crystallization, the crystallization was completed, filtration was performed, the filter cake was rinsed with an appropriate amount of toluene, and dried under reduced pressure to give compound TM30 in a yield of 92.0% and a purity of 99.89%.

¹H-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);

¹³C-NMR(400MHz,DMSO-d₆):δ170.1,168.6,167.8,159.2,149.7,145.6,142.4,136.5,134.2,133.1,132.3,130.1(2C),128.2,119.3(2C),118.5,111.0,99.6,40.1,33.7,31.2.

MS(m/z):548.27[M+H]⁺;

In vitro cytotoxic Activity test on cancer cells

In order to investigate the ability of the target compounds synthesized in this experiment to inhibit tumor cell proliferation, we examined the cytotoxicity of the compounds of the present invention against four tumor cells, namely lymphoma cell line (Raji), breast cancer cell (MCF 7), leukemia cell (K562) and head and neck squamous carcinoma cell (HN 5). The assay used was the standard MTT assay.

The experimental method specifically comprises the following steps:

Four tumor cells of lymphoma cell lines, breast cancer cells, leukemia cells and head and neck squamous carcinoma cells (Shanghai and order biotechnology Co., ltd.) are treated by pancreatin digestion for 10min, liquid is discarded, 5% serum culture solution is used for blowing, the cell concentration is regulated to 300-400 cells/mu L, medicines are sequentially added, a blank group only containing the culture solution is reserved, and the culture is carried out in an incubator for 24h. Discarding supernatant, adding diluted target compounds with different concentrations into 96-well plate (Shanghai macrobiosciences, inc.), adding cell sap into control group, setting three auxiliary holes for each group, mixing, culturing for 36 hr, observing cell morphology change in different time period, adding MTT solution for color development, and culturing for 6 hr. The upper liquid was discarded, 100 mu LDMSO was dissolved to form a purple crystal, and the absorbance was measured by a 490nm microplate reader (HBS-1096A microplate reader, nanjde iron laboratory equipment Co., ltd.). The IC ₅₀ value (plotted as cell viability versus log dose) was determined by the formula cell inhibition (%) = (1-a experiment/a control) ×100%.

In vitro toxicity test results of the compounds TM 1-8 on proliferation inhibition of four tumor cells, namely lymphoma cell line (Raji), breast cancer cell (MCF 7), leukemia cell (K562) and head and neck squamous carcinoma cell (HN 5), are shown in Table 1.

Part of the compounds of table 1 were tested for tumor cytotoxicity (IC ₅₀,μM)^a

Note that the values in table a are the average of 3 experiments, and the drug action time is 36h

The novel antitumor active compounds TM 1-30 provided by the invention have higher growth inhibition activities on four tumor cells, namely lymphoma cell lines (Raji), breast cancer cells (MCF 7), leukemia cells (K562) and head and neck squamous carcinoma cells (HN 5), the inhibition rates of the compounds TM10 on the lymphoma cell lines (Raji) and the leukemia cells (K562) reach the nM level, the inhibition rates of the compounds TM19 on the MCF7 reach the nM level, the inhibition rates of the compounds TM26 on the Raji and the MCF7 reach the nM level, and the inhibition rates of the three compounds on the four tumor cells are obviously higher than those of the control compound ibrutinib, and the inhibition activity and the selectivity of the compounds TM25 on the MCF7 cells are superior to those of the other three cells.

The compound TM10 also has good physical property, water solubility reaching 0.115mg/ml, and good metabolism in mice, and bioavailability reaching 96.8%.

Claims

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

2. A compound I The preparation method is characterized by comprising the following steps:

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, -NR ₂ R ₃ ; or one or more independent groups of substituted aryl, substituted heteroaryl, substituted C _3-6 cycloalkyl, substituted C _2-6 heterocycloalkyl, substituted aryloxy, substituted C _1-6 alkoxy, 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 heterocycloalkyl. _{R 2} is independently selected from H or C _1-4 alkyl;

_R3 is independently selected from one or more of H, _C1-4 alkyl, _C3-6 cycloalkyl, _C1-3 haloalkyl, aryl, heteroaryl, _C1-6 alkoxy, _C1-6 alkylamino, substituted _C2-6 heterocycloalkyl, halogen-substituted _C2-6 heterocycloalkyl, and _C2-6 heterocycloalkyl;

Z is independently selected from C _1-4 alkyl or aryl.

3. The method for preparing pyrazole-4,6-dione compounds according to claim 2, characterized in that the reaction step comprises:

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 pyrazole-4,6-dione compounds according to claim 2, characterized in that the specific reaction steps 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, 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 pyrazole-4,6-dione compounds according to claim 4, characterized in that the base in step 3 is one or two of sodium hydride, potassium hydroxide, sodium hydroxide, and lithium hydroxide.

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

9. The method for preparing pyrazole-4,6-dione compounds according to claim 4, characterized in that the organic solvent 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 optionally a pharmaceutically acceptable carrier, excipient, adjuvant, auxiliary material or diluent.

11. Use of the pyrazole-4,6-dione compound according to claim 1, characterized in that it is used as an active ingredient in the preparation of a drug for treating lymphoma, breast cancer, leukemia and head and neck squamous cell carcinoma.