Derivative of triptolide and triptonide, and synthesis method therefor and use thereof

Abstract

Disclosed in the present invention are a derivative of triptolide and triptonide, and a synthesis method therefor and the use thereof. The derivative of the present invention has the structure as shown in general formula I, and the definition of each substituent is as described in the description and claims. The derivative of triptolide and triptonide of the present invention exhibits a high anti-tumor activity, and good development and application prospects.

Description

Derivatives of triptolide and triptolide, their synthesis methods and applications Technical Field

[0001] This invention belongs to the fields of organic chemistry, medicinal chemistry and natural product chemistry, and specifically relates to a derivative of triptolide and triptolide ketone, its synthesis method and application. Background Technology

[0002] Tripterygium wilfordii Hook.f is a traditional Chinese herbal medicine with anti-inflammatory, antirheumatic, and analgesic activities.

[0003] Among the many natural products of Tripterygium wilfordii, triptolide is recognized as the main active ingredient. Tripterygium wilfordii, also known as triptolide lactone, is mainly extracted from the leaves and roots of Tripterygium wilfordii. It possesses significant anti-tumor, immunosuppressive, anti-inflammatory, and anti-male fertility activities, attracting widespread attention from medicinal chemistry and pharmacology researchers. However, studies have also found that triptolide has significant toxicity and various adverse reactions. Structural modification of triptolide to obtain highly effective and low-toxicity triptolide lactone derivatives, enabling them to exert anti-tumor and other biological activities, has received increasing attention in recent years.

[0004] Tripterygium wilfordii contains a rosinane diterpene structure with three epoxy groups and an α,β-unsaturated five-membered lactone ring. Structural modifications of triptolide mainly focus on the modification of the C14-hydroxyl group, the epoxy group, and the unsaturated lactone ring.

[0005] Currently, there are no marketed drugs specifically targeting the structural modification of triptolide. Triptolide-related derivatives or prodrug molecules typically exhibit lower biological activity than triptolide itself, highlighting the urgent need to develop more active triptolide derivatives for applications such as anti-tumor therapy. Summary of the Invention

[0006] The purpose of this invention is to provide triptolide and triptolide ketone derivatives modified with a five-membered lactone ring, their synthesis methods, and applications.

[0007] In a first aspect, the present invention provides a compound as shown in Formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph, or prodrug thereof.

[0008] Among them, CR3R4 is CHOH or C=O;

[0009] R1 and R2 are different, and are each independently selected from the following group: C1-C8 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C6-C10 aryl, -C1-C4 alkylene-C6-C10 aryl, 3-8 membered heterocyclic or 4-8 membered heteroaryl; the above groups are optionally substituted by one or more groups selected from the following group: halogen, -OH, NR a R b CN, COOH, unsubstituted or halogenated C1-C8 alkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C1-C8 alkoxy, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogenated C2-C6 alkynyl, unsubstituted or halogenated C6-C10 aryl, unsubstituted or halogenated 4-8 membered heteroaryl, unsubstituted or halogenated 4-8 membered heterocyclic; wherein, R a R b Each is independently selected from: H, C1-C8 alkyl groups.

[0010] In another preferred embodiment, R1 and R2 are different and are each independently selected from the group consisting of: C1-C6 alkyl, C3-C6 cycloalkyl, C2-C4 alkenyl, C2-C4 ynyl, C6-C10 aryl, -C1-C4 alkylene-C6-C10 aryl, 5-8 membered heterocyclic or 5-8 membered heteroaryl; the above groups are optionally substituted by one or more groups selected from the group consisting of: halogen, -OH, NR. a R b CN, COOH, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C3-C6 cycloalkyl, unsubstituted or halogenated C1-C6 alkoxy, unsubstituted or halogenated C2-C4 alkenyl, unsubstituted or halogenated C2-C4 alkynyl, unsubstituted or halogenated C6-C10 aryl, unsubstituted or halogenated 5-8 membered heteroaryl, unsubstituted or halogenated 5-8 membered heterocyclic; wherein, R a R b Each is independently selected from: H, C1-C6 alkyl groups.

[0011] In another preferred embodiment, R1 and R2 are different and are each independently selected from the group consisting of: C1-C4 alkyl, C3-C6 cycloalkyl, C2-C4 alkenyl, C2-C4 alkynyl, phenyl, -C1-C4 alkylene-phenyl, 5-7 membered heterocyclic or 5-7 membered heteroaryl; the above groups are optionally substituted by one or more groups selected from the group consisting of: halogen, -OH, NR. a R bCN, COOH, unsubstituted or halogenated C1-C4 alkyl, unsubstituted or halogenated C3-C6 cycloalkyl, unsubstituted or halogenated C1-C4 alkoxy, unsubstituted or halogenated C2-C4 alkenyl, unsubstituted or halogenated C2-C4 alkynyl, unsubstituted or halogenated phenyl, unsubstituted or halogenated 5-7 membered heteroaryl, unsubstituted or halogenated 5-7 membered heterocyclic; wherein, R a R b Each is independently selected from: H, C1-C4 alkyl groups.

[0012] In another preferred embodiment, R1 is selected from the group consisting of: C6-C10 aryl, C3-C6 cycloalkyl, 5-8 membered heterocyclic, or 5-8 membered heteroaryl; the above groups are optionally substituted by one or more groups selected from the group consisting of: halogen, -OH, NR. a R b , COOH, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkoxy; wherein, R a R b Each is independently selected from: H, C1-C6 alkyl groups.

[0013] In another preferred embodiment, R1 is selected from the group consisting of phenyl, C3-C6 cycloalkyl, and 5-6 heteroaryl groups; the above groups are optionally substituted by one or more groups selected from the group consisting of halogen, unsubstituted or halogenated C1-C6 alkyl, and C1-C4 alkoxy groups.

[0014] In another preferred embodiment, R2 is selected from the group consisting of: 5-8-membered heteroaryl, C1-C6 alkyl, C6-C10 aryl, and C3-C6 cycloalkyl; the above groups are optionally substituted by one or more groups selected from the group consisting of: halogen, -OH, and NR. a R b CN, COOH, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkoxy; wherein, R a R b Each is independently selected from: H, C1-C6 alkyl groups.

[0015] In another preferred embodiment, R2 is selected from the group consisting of: phenyl, C1-C4 alkyl, C3-C6 cycloalkyl, 5-6 heteroaryl; the above groups are optionally substituted by one or more groups selected from the group consisting of: halogen, NR a R b Unsubstituted or halogenated C1-C4 alkyl or C1-C4 alkoxy groups; wherein, R a R b Each is independently selected from: H, C1-C4 alkyl groups.

[0016] In another preferred embodiment, the compound is selected from the group consisting of:

[0017] A second aspect of the present invention provides a method for preparing the compound described in the first aspect, the method comprising the steps of:

[0018] (a) The compound of formula II reacts with the first acylation reagent to give the compound of general formula III: wherein the first acylation reagent is R1COCl, R1COBr or R1COOCOR1;

[0019] (b) The compound of general formula III is reacted with a second acylation reagent to give a compound of formula I, wherein the second acylation reagent is R2COCl, R2COBr, or R2COOCOR2;

[0020] In each formula, R1, R2, R3 and R4 are defined as described above.

[0021] In another preferred embodiment, in step (a), the reaction is carried out in an organic solvent in the presence of an organometallic base, wherein,

[0022] The organometallic base is selected from the group consisting of: lithium diisopropylamino (LDA), n-butyllithium (n-BuLi), tert-butyllithium (t-BuLi), lithium hexamethyldisilamide (LiHMDS), sodium hexamethyldisilamide (NaHMDS), potassium hexamethyldisilamide (KHMDS), 2,2,6,6-tetramethylpiperidinyllithium, and 2,2,6,6-tetramethylpiperidinyllithium chloride and magnesium chloride.

[0023] The organic solvent is selected from the group consisting of tetrahydrofuran, diethyl ether, 2-methyltetrahydrofuran, or a mixture of two or more solvents.

[0024] In another preferred embodiment, in step (b), the reaction is carried out in an organic solvent in the presence of a base, wherein the base is selected from: triethylamine, diisopropylamine, N,N-diisopropylethylamine, pyridine, and piperidine;

[0025] The organic solvent is selected from the group consisting of ethyl acetate, isopropyl acetate, dichloromethane, chloroform, tetrahydrofuran, 2-methyltetrahydrofuran, diethyl ether, acetonitrile, 1,4-dioxane, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, or a mixture of two or more solvents.

[0026] In another preferred embodiment, in step (a), after the reaction is completed, an acidic aqueous solution is added for quenching to obtain a compound of general formula III, wherein the acidic aqueous solution is prepared by dissolving hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid, formic acid, acetic acid, oxalic acid, maleic acid, fumaric acid, succinic acid or citric acid in water.

[0027] A third aspect of the present invention provides a pharmaceutical composition comprising:

[0028] The compound described in the first aspect, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph, or prodrug thereof; and

[0029] Pharmaceutically acceptable carrier.

[0030] A fourth aspect of the present invention provides the use of the compound described in the first aspect, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof, or a pharmaceutical composition described in the third aspect, for the preparation of a medicament for treating tumors.

[0031] In another preferred embodiment, the tumor is selected from the group consisting of: leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, squamous cell carcinoma of the lung, adenocarcinoma of the lung, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, cervical cancer, ovarian cancer, intestinal cancer, nasopharyngeal carcinoma, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, and oral cancer.

[0032] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0033] The inventors of this application, through extensive and in-depth research, modified the five-membered lactone rings of triptolide and triptolide ketone to obtain a series of derivatives of triptolide and triptolide ketone with antitumor activity. Based on this, the present invention was completed.

[0034] the term

[0035] Unless otherwise defined, all technical terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent applications, and publications cited in this document are incorporated herein in their entirety through reference.

[0036] Certain chemical groups defined herein are preceded by simplified symbols to indicate the total number of carbon atoms present in the group. For example, C1-6 alkyl refers to an alkyl group having a total of 1 to 6 carbon atoms as defined below. And so on. "3-8" refers to a group having 3, 4, 5, 6, 7, or 8 ring atoms, and so on. The total number of carbon atoms in the simplified symbols does not include carbons that may be present in substituents of the group.

[0037] Except as otherwise specified, when used in the specification and claims of this application, the following terms shall have the following meanings.

[0038] In this application, the term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0039] In this application, as a group or part of other groups (e.g., in halogen-substituted alkyl groups), the term "alkyl" means a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, without unsaturated bonds, having, for example, 1 to 8 (preferably 1 to 6, more preferably 1 to 4) carbon atoms connected to the rest of the molecule by single bonds. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, heptyl, 2-methylhexyl, 3-methylhexyl, octyl, etc.

[0040] "Alkylene" refers to a straight-chain or branched saturated aliphatic group, i.e., a divalent hydrocarbon group, having a specified number of carbon atoms and being attached to at least two other groups. The two groups attached to the alkylene group can be the same or different atoms attached to the alkylene group. Representative alkylene groups include, but are not limited to, methylene, ethylene, propylene, isopropylene, butylene, isobutylene, and sec-butylene.

[0041] In this application, as part of a group or other group, the term "alkenyl" means a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing at least one double bond, having, for example, 2 to 6 (preferably 2 to 4) carbon atoms connected to the rest of the molecule by single bonds, such as, but not limited to, vinyl, propenyl, allyl, but-1-alkenyl, but-2-alkenyl, etc.

[0042] In this application, as part of a group or other group, the term "alkynyl" means a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing at least one triple bond and optionally one or more double bonds, having, for example, 2 to 6 (preferably 2 to 4) carbon atoms and connected to the rest of the molecule by single bonds, such as, but not limited to, ethynyl, propynyl, butynyl, etc.

[0043] In this application, as a group or part of other groups, the term "cycloalkyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon group consisting only of carbon and hydrogen atoms. It may include fused ring systems, bridged ring systems, or spirocyclic systems, preferably having 3 to 8 carbon atoms, more preferably having 3 to 6 carbon atoms, and is either saturated or unsaturated and can be linked to the rest of the molecule via a single bond through any suitable carbon atom. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, and cyclohexadienyl.

[0044] In this application, as part of a group or other group, the term "heterocyclic group" means a stable 3- to 8-membered non-aromatic cyclic group consisting preferably of 2 to 7 carbon atoms and preferably 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen, and sulfur. Unless otherwise specifically indicated in this specification, a heterocyclic group can be a monocyclic, bicyclic, tricyclic, or more ring system, which may include fused ring systems, bridged ring systems, or spirocyclic systems; the nitrogen, carbon, or sulfur atoms in the heterocyclic group may optionally be oxidized; the nitrogen atom may optionally be quaternized; and the heterocyclic group may be partially or fully saturated. The heterocyclic group may be connected to the remainder of the molecule via a carbon atom or heteroatom and by a single bond. In heterocyclic groups containing fused rings, one or more rings may be aryl or heteroaryl as defined below, provided that the connection point with the remainder of the molecule is a non-aromatic ring atom. For the purposes of this invention, the heterocyclic group is preferably a stable 3- to 8-membered non-aromatic monocyclic, bicyclic, bridged, or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur; more preferably, it is a stable 4- to 7-membered non-aromatic monocyclic, bicyclic, bridged, or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur. Examples of heterocyclic groups include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, nitrogen-containing heterocyclic butyl, pyranyl, dioxolaneyl, imidazoalkyl, pyrazolyl, etc.

[0045] In this application, as a group or part of other groups, the term "aryl" refers to a conjugated hydrocarbon ring system group having 6 to 10 carbon atoms (preferably 6 carbon atoms). For the purposes of this invention, the aryl group can be a monocyclic or bicyclic ring system, and can also be fused with cycloalkyl or heterocyclic groups as defined above, provided that the aryl group is connected to the rest of the molecule via single bonds through atoms on the aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, etc.

[0046] In this application, as part of a group or other group, the term "heteroaryl" refers to a 4- to 8-membered conjugated cyclic group having 1 to 7 carbon atoms (preferably 1 to 5 carbon atoms) and 1 to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specifically indicated in this specification, a heteroaryl group may be a monocyclic or bicyclic cyclic system and may be fused with a cycloalkyl or heterocyclic group as defined above, provided that the heteroaryl group is connected to the rest of the molecule via a single bond through an atom on the aromatic ring. The nitrogen, carbon, or sulfur atom in the heteroaryl group may optionally be oxidized; the nitrogen atom may optionally be quaternized. For the purposes of this invention, the heteroaryl group is preferably a stable 5- to 7-membered aromatic group containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur, more preferably a stable 5- to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur, or a 5- to 6-membered aromatic group containing 1 to 2 heteroatoms selected from nitrogen, oxygen, and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzimidazolyl, benzopyrazolyl, indolyl, furanyl, pyrroleyl, triazolyl, tetrazolyl, etc.

[0047] In this application, "optional" or "optionally" means that the event or condition described below may or may not occur, and the description includes both the occurrence and non-occurrence of the event or condition. For example, "optionally substituted aryl" means that the aryl group is substituted or not substituted, and the description includes both substituted and unsubstituted aryl groups.

[0048] In this invention, the substitution can be monosubstituted or polysubstituted, and the polysubstituted can be disubstituted, trisubstituted, tetrasubstituted, or pentasubstituted. Disubstituted means having two substituents, and so on.

[0049] "Stereoisomers" are compounds composed of identical atoms bonded by the same bonds, but with different three-dimensional structures. This invention will cover various stereoisomers and mixtures thereof.

[0050] When the compounds of the present invention contain alkene double bonds, unless otherwise stated, the compounds of the present invention are intended to contain E- and Z-geometric isomers.

[0051] "Tautomer" refers to an isomer formed when a proton is transferred from one atom of a molecule to another atom of the same molecule. All tautomer forms of the compounds of this invention are also included within the scope of this invention.

[0052] The compounds of the present invention, or pharmaceutically acceptable salts thereof, may contain one or more chiral carbon atoms, and thus may produce enantiomers, diastereomers, and other stereoisomers. Each chiral carbon atom may be defined as (R)- or (S)- based on stereochemistry. The present invention aims to include all possible isomers, as well as their racemic and optically pure forms. The preparation of the compounds of the present invention may select racemic, diastereomer, or enantiomer as starting materials or intermediates. Optically active isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

[0053] Conventional techniques for preparing / separating individual isomers include chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high-performance liquid chromatography.

[0054] In this application, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0055] "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the bioavailability of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobroms, sulfates, nitrates, and phosphates; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetate, trifluoroacetate, propionates, hexanoates, octanoates, decanoates, undecenoates, glycolates, gluconates, lactates, sebates, adipates, glutarate, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartate, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginates, ascorbic acid salts, salicylates, 4-aminosalicylic acid salts, and naphthalene disulfonates. These salts can be prepared using methods known in this field.

[0056] "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that retain the bioavailability of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, the following: primary amines, secondary amines, and tertiary amines; substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in the art.

[0057] "Polymorphic compounds" refer to different solid crystalline phases resulting from the presence of two or more different molecular arrangements in the solid state of certain compounds of the present invention. Some compounds of the present invention may exist in more than one crystal form, and the present invention aims to include various crystal forms and mixtures thereof.

[0058] Typically, crystallization produces solvates of the compounds of this invention. The term "solvate" as used in this invention refers to an aggregate comprising one or more molecules of the compound of this invention and one or more solvent molecules. The solvent may be water, in which case the solvate is a hydrate. Alternatively, the solvent may be an organic solvent. Therefore, the compounds of this invention can exist as hydrates, including monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, etc., and corresponding solvated forms. The compounds of this invention can form true solvates, but in some cases, they may also remain only as indeterminate water or a mixture of water and a portion of indeterminate solvent. The compounds of this invention can react in a solvent or precipitate or crystallize from a solvent. The solvates of the compounds of this invention are also included within the scope of this invention.

[0059] This invention also includes prodrugs of the aforementioned compounds. In this application, the term "prodrug" refers to a compound that can be converted into the bioactive compound of this invention under physiological conditions or by solvent decomposition. Therefore, the term "prodrug" refers to a pharmaceutically acceptable metabolic precursor of the compound of this invention. When administered to an individual in need, the prodrug may be inactive but is converted in vivo into the active compound of this invention. Prodrugs are typically rapidly converted in vivo to produce the parent compound of this invention, for example, through hydrolysis in the blood. Prodrug compounds generally offer advantages such as solubility, tissue compatibility, or sustained release in mammalian organisms. Prodrugs include known amino and carboxyl protecting groups.

[0060] In this application, "pharmaceutical composition" refers to a formulation of the compounds of the present invention with a medium generally accepted in the art for delivering bioactive compounds to mammals (e.g., humans). This medium includes pharmaceutically acceptable carriers. The purpose of the pharmaceutical composition is to facilitate administration to the organism, thereby promoting the absorption of the active ingredient and the exertion of its bioactivity.

[0061] As used herein, the term "pharmaceutically acceptable" means a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compounds of the present invention and is relatively non-toxic, i.e., that the substance can be administered to an individual without causing an adverse biological reaction or interacting adversely with any component contained in the composition.

[0062] In this application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, flow aid, sweetener, diluent, preservative, dye / coloring agent, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that is permitted by the relevant government regulatory authority to be acceptable for human or animal use.

[0063] The "tumor" and "diseases related to abnormal cell proliferation" mentioned in this invention include, but are not limited to, leukemia, gastrointestinal stromal tumors, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, squamous cell carcinoma of the lung, adenocarcinoma of the lung, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, cervical cancer, ovarian cancer, colorectal cancer, nasopharyngeal carcinoma, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, oral cancer, and other diseases.

[0064] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions (such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989)) or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0065] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0066] Example 1: Synthesis of Compound 1

[0067] Step 1) Synthesis of Compound 1-1

[0068] Under nitrogen protection, triptonide (300 mg, 0.84 mmol) was added to a reaction flask, followed by 15 mL of anhydrous tetrahydrofuran. The mixture was cooled to approximately -60 °C, and 3.3 mL (3.3 mmol) of a 1 mol / L tetramethylpiperidinyl lithium chloride magnesium chloride complex tetrahydrofuran solution was slowly added dropwise. After stirring at this temperature for 30 min, 230 mg (1.64 mmol) of benzoyl chloride was slowly added dropwise. Once the addition was complete, the reaction was maintained at -60 °C for 3 h. After the reaction was complete, dilute hydrochloric acid was added to quench the reaction, followed by extraction with ethyl acetate. The concentrate was then purified by silica gel column chromatography to obtain compound 1-1 (300 mg, 77% yield).

[0069] LCMS(m / z): 463.1 [M+1] + .

[0070] 1H NMR(400MHz, CDCl3)δ8.05(d,J=7.1Hz,2H),7.65(t,J=7.4Hz,1H),7.52(t,J=7.7Hz,2H), 5.90(s,1H),4.06(d,J=2.9Hz,1H),3.84(d,J=2.8Hz,1H),3.36(d,J=5.5Hz,1H),3.06(dd, J1=12.3Hz, J2=6.2Hz,1H),2.43–2.36(m,2H),2.29–2.21(m,2H),1.98–1.91(m,1H),1.62– 1.58(m,1H),1.44–1.36(m,1H),1.08(s,3H),0.98(d,J=6.8Hz,3H),0.89(d,J=7.0Hz,3H).

[0071] 13 C NMR (100MHz, CDCl3) δ196.96,192.02,171.07,159.54,134.65,134.44,129.59,129.59,128.96,128.96,127.41, 81.67,66.45,65.14,60.95,60.51,58.91,56.07,41.05,35.49,30.06,25.89,23.47,18.03,17.41,16.35,14.01.

[0072] Step 2) Synthesis of Compound 1

[0073] Under nitrogen protection, compound 1-1 (40 mg, 0.086 mmol), isonicotinyl chloride hydrochloride (88 mg, 0.49 mmol), and 2 mL of dichloromethane were added to a reaction flask. The mixture was cooled to below -60 °C, and diisopropylethylamine (40 mg, 0.31 mmol) was slowly added dropwise. The reaction was then carried out at -60 °C for approximately 0.5 h, and the reaction was monitored for completeness by LC-MS. The mixture was quenched with water, concentrated, and purified by preparative chromatography to give compound 1 (19 mg, 39% yield).

[0074] LCMS(m / z): 568.2 [M+1] + .

[0075] 1H NMR (400MHz, CDCl3) δ8.89 (s, 2H), 8.07 (d, J = 4.7Hz, 2H), 7.72 (dd, J1 = 7.7Hz, J2 = 2.0Hz, 2H) ,7.46–7.39(m,3H),3.93(d,J=2.8Hz,1H),3.73(d,J=2.7Hz,1H),2.75–2.68(m,2H),2.54(d d, J1=12.9Hz, J2=3.1Hz,1H),2.42–2.35(m,2H),2.33–2.26(m,1H),2.02–1.91(m,1H),1.61 –1.56(m,1H),1.23–1.18(m,1H),1.11(s,3H),0.93(d,J=6.8Hz,3H),0.87(d,J=6.9Hz,3H).

[0076] 13 C NMR (100MHz, CDCl3) δ196.51,167.61,163.56,151.45,151.45,149.71,142.38,135.27,132.93,131.22,130.38,129.89,128.94,128.94,1 28.15,128.15,123.08,123.08,66.62,65.12,60.68,60.03,58.81,5 5.62,41.20,36.39,30.05,26.14,24.51,18.04,17.87,16.68,15.37.

[0077] Example 2: Synthesis of Compound 2

[0078] Compound 1-1 (60 mg, 0.13 mmol), dimethylaminobenzoyl chloride (72 mg, 0.39 mmol), and 6 mL of dichloromethane were added to a reaction flask. The mixture was cooled to approximately 5 °C in an ice bath, and triethylamine (39 mg, 0.39 mmol) was slowly added dropwise. The reaction was allowed to proceed for approximately 0.5 hours, and LC-MS analysis confirmed that the starting material had reacted completely. The reaction was stopped by quenching with water, and the mixture was concentrated to obtain compound 2 (28 mg, 35% yield).

[0079] LCMS(m / z): 610.2 [M+1] + .

[0080] 1H NMR (400MHz, DMSO-d6) δ7.90–7.84(m,4H),7.64(t,J=7.4Hz,1H),7.54(t,J=7.7Hz,2H ),6.79(d,J=9.2Hz,2H),4.30(d,J=2.9Hz,1H),4.19(d,J=2.8Hz,1H),3.52(d,J=5.7Hz ,1H),3.39-3.35(m,1H),3.11–3.07(m,1H),3.06(s,6H),2.37–2.26(m,3H),1.75-1.6 9(m,1H),1.49–1.38(m,2H),1.07(s,3H),0.93(d,J=6.7Hz,3H),0.85(d,J=7.0Hz,3H).

[0081] 13 C NMR(100MHz,DMSO-d6)δ197.99,182.42,161.83,154.41,147.95,141.35,1 37.24,136.09,132.67,132.29,132.29,129.28,129.28,128.40,128.40,11 1.12,111.12,110.91,107.62,66.01,65.24,60.89,60.35,58.96,55.68,39.52,39.52,39.10,35.16,29.27,25.51,22.32,17.79,16.16,14.56,14.14.

[0082] Example 3: Synthesis of Compound 3

[0083] Under nitrogen protection, compound 1-1 (54 mg, 0.12 mmol) was dissolved in 5 mL of dichloromethane, cooled to ~5 °C in an ice bath, and acetyl chloride (21 mg, 0.27 mmol) and triethylamine (30 mg, 0.30 mmol) were slowly added dropwise. The reaction was then carried out at room temperature for 1 h, and the reaction was monitored for completeness by LC-MS. The mixture was quenched with water, concentrated, and purified by preparative chromatography to give compound 3 (23 mg, 39% yield).

[0084] LCMS(m / z): 505.0 [M+1] + .

[0085] 1H NMR (400MHz, CDCl3) δ7.90–7.88(m,2H),7.59–7.55(m,1H),7.47(t,J=7.7Hz,2H),4.04 (d,J=2.9Hz,1H),3.82(d,J=2.8Hz,1H),3.53–3.47(m,1H),3.44–3.43(m,1H),3.18(dd, J1=12.2Hz, J2=5.4Hz,1H),2.46–2.43(m,1H),2.41–2.36(m,2H),2.30(s,3H),1.83–1. 76(m,2H),1.37–1.32(m,1H),1.18(s,3H),0.99(d,J=6.8Hz,3H),0.91(d,J=7.0Hz,3H).

[0086] Example 4: Synthesis of Compound 4

[0087] Compound 1-1 (20 mg, 0.043 mmol) was reacted with cyclohexaneformyl chloride (19 mg, 0.13 mmol) and triethylamine (13 mg, 0.13 mmol) according to Example 2, and after purification, compound 4 (8 mg, 32% yield) was obtained.

[0088] LCMS(m / z): 573.2 [M+1] + .

[0089] 1 H NMR(400MHz, CDCl3) δ7.66(d,J=7.8Hz,2H),7.43–7.37(m,3H),4.01(d,J=2.9Hz,1H),3.83 (d,J=2.8Hz,1H),3.44(d,J=6.1Hz,1H),3.07–3.00(m,1H),2.67–2.63(m,1H),2.55–2.49(m ,1H),2.41–2.34(m,3H),2.14(t,J=15.2Hz,2H),2.02–1.94(m,1H),1.86–1.76(m,3H),1.62 –1.50(m,5H),1.34–1.31(m,2H),1.15(s,3H),0.98(d,J=6.9Hz,3H),0.91(d,J=6.9Hz,3H).

[0090] Example 5: Synthesis of Compound 5

[0091] Compound 1-1 (30.1 mg, 0.065 mmol) and p-methoxybenzoyl chloride (14.8 mg, 0.087 mmol) and triethylamine (8.6 mg, 0.085 mmol) were reacted according to Example 2, and after purification, compound 5 (35 mg, 90% yield) was obtained.

[0092] LCMS(m / z): 597.0 [M+1] + .

[0093] 1 H NMR (400MHz, CDCl3) δ8.10–8.07(m,2H),7.94–7.92(m,2H),7.57–7.53(m,1H),7.49–7.45(m,2H),6.99–6.96(m,2H),4.05(d,J=2.5Hz,1H),3. 89(s,3H),3.83(d,J=2.8Hz,1H),3.58–3.51(m,1H),3.46(d,J=5.8Hz,1H),3.22(dd,J1=12.1Hz,J2=5.4Hz,1H),2.49–2.40(m,3H),1.83(dd,J 1=15.2Hz,J 2=12.2Hz,1H),1.42–1.34(m,1H),1.34–1.30(m,1H),1.22(s,3H),0.99(d,J=6.8Hz,3H),0.91(d,J=7.0Hz,3H).

[0094] Example 6: Synthesis of Compound 6

[0095] Compound 1-1 (40 mg, 0.086 mmol) was reacted with p-trifluoromethylbenzoyl chloride (24.1 mg, 0.12 mmol) and triethylamine (11.7 mg, 0.12 mmol) according to Example 2, and after purification, compound 6 (27 mg, 49% yield) was obtained.

[0096] LCMS(m / z): 635.0 [M+1] + .

[0097] 1H NMR(400MHz, CDCl3)δ8.26(d,J=8.1Hz,2H),7.94–7.91(m,2H),7.79(d,J=8.3Hz,2H),7.59–7.54(m, 1H),7.49–7.45(m,2H),4.06(d,J=2.9Hz,1H),3.83(d,J=2.8Hz,1H),3.55–3.49(m,1H),3.46(d,J=5 .9Hz,1H),3.23(dd,J1=12.1Hz,J2=5.4Hz,1H),2.47-2.39(m,3H),1.83(dd,J1=15.0Hz,J2=12.2Hz, 1H),1.62–1.57(m,1H),1.42–1.33(m,1H),1.22(s,3H),0.99(d,J=6.8Hz,3H),0.91(d,J=7.0Hz,3H).

[0098] Example 7 Synthesis of Compound 7

[0099] Step 1: Synthesis of Compound 7-1

[0100] Tripterygium lactone (60 mg, 0.17 mmol), 2,2,6,6-tetramethylpiperidinyl lithium chloride magnesium chloride complex tetrahydrofuran solution (0.7 mL, 0.70 mmol), 4-methoxybenzoyl chloride (57 mg, 0.33 mmol), were synthesized according to the first step of Example 1, and purified to obtain compound 7-1 (33 mg, 40% yield).

[0101] LCMS(m / z): 493.0 [M+1] + .

[0102] Step 2: Synthesis of Compound 7

[0103] Compound 7-1 (30 mg, 0.061 mmol), benzoyl chloride (13 mg, 0.093 mmol), and triethylamine (25 mg, 0.25 mmol) were synthesized according to the method in Example 2. The compounds were separated and purified by preparative chromatography to obtain compound 7 (23 mg, yield 63%).

[0104] LCMS(m / z): 597.2 [M+1] + .

[0105] 1H NMR (400MHz, CDCl3) δ8.16–8.14(m,2H),7.99–7.95(m,2H),7.70–7.66(m,1H),7.52(t ,J=7.7Hz,2H),6.98–6.94(m,2H),4.05(d,J=2.9Hz,1H),3.86(s,3H),3.83(d,J=4.0Hz 1H), 3.53–3.44 (m, 2H), 3.22 (dd, J1=12.1, J2=5.2Hz, 1H), 2.47-2.40 (m, 3H), 1.81 (dd, J1=14.8Hz, J2=12. 1Hz,1H),1.59–1.55(m,1H),1.40-1.34(m,1H),1.21(s,3H),0.99(d,J=7.0Hz,3H),0.91(d,J=6.9Hz,3H).

[0106] Example 8: Synthesis of Compound 8

[0107] Synthesis of compound 8-1 in step one

[0108] Tripterygium wilfordii (1.0 g, 2.8 mmol), 2,2,6,6-tetramethylpiperidinyl lithium chloride magnesium chloride complex tetrahydrofuran solution (11.1 mL, 11.1 mmol), benzoyl chloride (0.8 g, 5.7 mmol), were synthesized according to the first step of Example 1, and purified to give compound 8-1 (0.69 g, 53% yield).

[0109] LCMS(m / z): 465.2 [M+1] + .

[0110] 1 H NMR (400MHz, CDCl3) δ8.04(d,J=8.0Hz,2H),7.66(t,J=8.0Hz,1H),7.52(t,J=8.0Hz,2H),5.89- 5.88(m,1H),3.92(d,J=4.0Hz,1H),3.52(d,J=4.0Hz,1H),3.40(s,1H),3.32(d,J=4.0Hz,1H),2 .97-2.93(m,1H),2.71(br,1H),2.40-2.34(m,1H),2.29-2.18(m,3H),1.94(t,J=12.0Hz,1H),1 .59-1.55(m,2H),1.35-1.28(m,1H),1.14(s,3H),1.01(d,J=8.0Hz,3H),0.89(d,J=8.0Hz,3H).

[0111] Step 2: Synthesis of Compound 8

[0112] Compound 8-1 (100 mg, 0.22 mmol), isonicotinic acid chloride hydrochloride (113 mg, 0.63 mmol), and diisopropylethylamine (278 mg, 2.16 mmol) were purified to obtain compound 8 (64 mg, 52% yield) according to the synthetic method in step 2 of Example 1.

[0113] LCMS(m / z): 570.2 [M+1] + ;

[0114] 1 H NMR (400MHz, CDCl3) δ8.94 (s, 2H), 8.04 (d, J = 6.0Hz, 2H), 7.70 (dd, 7.9Hz, J2 = 1.9Hz, 2H),7.45–7.40(m,3H),3.79(d,J=3.1Hz,1H),3.40(d,J=3.1Hz,1H),3.03(s,1H),2. 63–2.51(m,4H),2.38–2.34(m,2H),2.20–2.14(m,1H),1.96–1.89(m,1H),1.57–1.53 (m,1H),1.15(s,3H),1.14–1.09(m,1H),0.94(d,J=7.0Hz,3H),0.84(d,J=6.9Hz,3H).

[0115] 13 C NMR (100MHz, CDCl3) δ167.63,163.34,151.28,151.28,149.92,142.24,135.20,132.86,131.26,130.09,129.60,128.71,128.71,127.98, 127.98,123.10,123.10,72.88,65.80,65.51,60.23,60.08,56.27,5 3.88,40.85,36.78,29.16,28.05,24.67,17.76,17.65,16.76,14.90.

[0116] Example 9: Synthesis of Compound 9

[0117] Under nitrogen protection, compound 8-1 (20 mg, 0.043 mmol), dimethylaminobenzoyl chloride (24 mg, 0.13 mol), and 2 mL of dichloromethane were added to a reaction flask. The mixture was cooled to below -60 °C, and triethylamine (13 mg, 0.13 mmol) was slowly added dropwise. The reaction was then carried out at -60 °C for 0.5 h. LC-MS monitoring showed that a small amount of the starting material remained. The mixture was quenched with water, concentrated, and purified by preparative chromatography to obtain compound 9 (7 mg, 27% yield).

[0118] LCMS(m / z): 612.3 [M+1] + ;

[0119] 1 H NMR (400MHz, DMSO-d6) 1 H NMR (400MHz, DMSO-d6) δ7.87–7.80(m,4H),7.63–7.59(m,1H),7.51(t,J=7.7Hz,2H),6.76(d,J=9.2H z,2H),4.74(d,J=7.4Hz,1H),3.90(d,J=3.2Hz,1H),3.56(d,J=3.0Hz,1H),3.39–3.34(m,2H),3.25–3 .18(m,1H),3.03(s,6H),2.90–2.86(dd,J1=12.3Hz,J2=5.7Hz,1H),2.30–2.26(m,2H),2.19–2.12(m, 1H),1.66–1.59(m,1H),1.35–1.25(m,2H),1.08(s,3H),0.92(d,J=6.9Hz,3H),0.78(d,J=6.9Hz,3H).

[0120] Example 10: Synthesis of Compound 10

[0121] Compound 8-1 (20 mg, 0.043 mmol) was reacted with cyclopropaneformyl chloride (13 mg, 0.12 mmol) and triethylamine (13 mg, 0.13 mmol) according to Example 2, and after purification, compound 10 (8 mg, 35% yield) was obtained.

[0122] LCMS(m / z): 533.2 [M+1] + ;

[0123] 1H NMR (400MHz, CDCl3) δ7.89(d,J=7.0Hz,2H),7.56(t,J=7.4Hz,1H),7.47(t,J=7.6Hz,2H),3.89(d,J= 3.2Hz,1H),3.50–3.38(m,4H),3.05(dd,J1=12.1Hz,J1=5.7Hz,1H),2.73(d,J=10.3Hz,1H),2.35(dd ,J1=9.2Hz,J1=3.6Hz,2H),2.30–2.23(m,1H),1.83–1.76(m,1H),1.55–1.50(m,1H),1.26–1.24(m,1 H),1.22(s,3H),1.20–1.17(m,2H),1.11–1.07(m,2H),1.03(d,J=7.0Hz,3H),0.89(d,J=6.9Hz,3H).

[0124] 13 C NMR (100MHz, CDCl3) δ183.31,170.57,147.22,141.95,137.70,136.23,132.43,129.80,129.80,128.17,128.17,107.51,73.58, 66.50,65.50,61.15,60.36,56.69,54.41,39.86,36.19,29.45,28.19,23.31,17.75,16.86,15.02,14.07,12.33,10.30,10.28.

[0125] Example 11: Synthesis of Compound 11

[0126] Synthesis of compound 11-1 in step one

[0127] Tripterygium wilfordii (100 mg, 0.28 mmol) and furanoyl chloride (70 mg, 0.54 mmol), along with 1.1 mL (1.1 mmol) of a tetrahydrofuran solution of a 2,2,6,6-tetramethylpiperidinyl lithium chloride magnesium chloride complex, were reacted according to the first step of Example 1. After purification, compound 11-1 (35 mg, yield 28%) was obtained.

[0128] LCMS(m / z): 455.1 [M+1] + ;

[0129] 1H NMR (400MHz, CDCl3) δ7.70 (d, J=1.7Hz, 1H), 7.49 (d, J=3.7Hz, 1H), 6.60 (dd, J1=3.7Hz, J2=1.7Hz, 1H), 5.63–5.62(m,1H),3.85(d,J=3.1Hz,1H),3.47(d,J=3.1Hz,1H),3.40(s,1H),3.36(d,J=5.6Hz,1H),2.7 0–2.67(m,1H),2.59–2.54(m,1H),2.38–2.32(m,1H),2.26–2.19(m,2H),2.00–1.92(m,1H),1.56–1.51( m,1H),1.22–1.16(m,1H),1.13(s,3H),1.10–1.05(m,1H),1.00(d,J=7.0Hz,3H),0.87(d,J=6.9Hz,3H).

[0130] The second step involves the synthesis of compound 11.

[0131] Compound 11-1 (20 mg, 0.044 mmol) was reacted with isonicotinic chloride hydrochloride (23 mg, 0.13 mmol) and diisopropylethylamine (54 mg, 0.42 mmol) according to Example 1, and purified to give compound 11 (7 mg, yield 28%).

[0132] LCMS(m / z): 560.2 [M+1] + ;

[0133] 1 H NMR(400MHz,Chloroform-d)δ8.92(s,2H),8.05(d,J=6.2Hz,2H),7.48(d,J=1.8Hz,1H),7.21(d,J=3.6H z,1H),6.59(dd,J1=3.6Hz,J2=1.8Hz,1H),3.77(d,J=3.1Hz,1H),3.39(d,J=2.2Hz,1H),3.04(s,1H),2. 71–2.64(m,2H),2.56(s,1H),2.47–2.32(m,3H),2.19–2.12(m,1H),1.93–1.86(m,1H),1.56–1.51(m,1H ),1.14(s,3H),1.12–1.06(dd,J1=11.4Hz,J2=6.3Hz,1H),0.93(d,J=7.0Hz,3H),0.82(d,J=6.9Hz,3H).

[0134] Example 12 Synthesis of Compound 12

[0135] Synthesis of Compound 12-1 (Step 1)

[0136] Tripterygium wilfordii (100 mg, 0.28 mmol) and cyclohexaneformyl chloride (79 mg, 0.54 mmol), along with 1.1 mL (1.1 mmol) of a tetrahydrofuran solution of a 2,2,6,6-tetramethylpiperidinyl lithium chloride magnesium chloride complex, were reacted according to the first step of Example 1. After purification, compound 12-1 (30 mg, yield 23%) was obtained.

[0137] LCMS(m / z): 471.2 [M+1] + ;

[0138] 1 H NMR (400MHz, CDCl3) δ5.20–5.19(m,1H),3.85(d,J=3.2Hz,1H),3.49(d,J=3.0Hz,1 H),3.42(s,1H),3.38(d,J=5.7Hz,1H),2.74–2.67(m,2H),2.56–2.51(m,1H),2.36– 2.12(m,3H),2.02–1.97(m,1H),1.91–1.84(m,1H),1.80-1.66(m,5H),1.56–1.49(m ,2H),1.40–1.20(m,5H),1.09(s,3H),1.01(d,J=7.0Hz,3H),0.87(d,J=6.9Hz,3H).

[0139] The second step involves the synthesis of compound 12.

[0140] Compound 12-1 (15 mg, 0.032 mmol) was reacted with isonicotinic chloride hydrochloride (17 mg, 0.096 mmol) and diisopropylethylamine (40 mg, 0.31 mmol) according to Example 1, and purified to give compound 12 (3 mg, 16% yield).

[0141] LCMS(m / z): 576.2 [M+1] + ;

[0142] 1H NMR (400MHz, CDCl3) δ8.90(s,2H),8.00(s,2H),3.76(d,J=3.1Hz,1H),3.39(d,J=3.0Hz,1 H),3.15-3.07(m,1H),2.95(s,1H),2.54–2.49(m,2H),2.44(d,J=6.2Hz,1H),2.34–2.22( m,3H),2.18–2.12(m,1H),2.03-2.01(m,1H),1.92–1.74(m,6H),1.52–1.45(m,2H),1.39– 1.33(m,2H),1.18–1.15(m,1H),1.08(s,3H),0.92(d,J=6.9Hz,3H),0.83(d,J=6.9Hz,3H).

[0143] Example 14: Detection of the in vitro antitumor activity of small molecule compounds

[0144] The tumor cells were ASPC-1 (human pancreatic cancer cells).

[0145] The cell sources and culture media are shown in Table 1 below.

[0146] Table 1 Tumor Cell Information Table

[0147] Tripterygium wilfordii (TP) was used as a positive control; the working concentrations of the test compounds were designed to be 2 μM, 0.67 μM, 0.22 μM, 0.074 μM, 0.025 μM, 0.008 μM, 0.0027 μM, 0.0009 μM, 0.0003 μM, and 0.0001 μM.

[0148] Experimental methods:

[0149] After tumor cells were resuscitated, they were resuspended in the appropriate culture medium, and the cell concentration was adjusted to a final seeding density of 6000 cells / plate. The plates were then incubated overnight in a CO2 incubator. The test drugs at various concentrations were then added, and the plates were incubated at 37°C in a 5% CO2 incubator for 72 hours. The inhibitory effect of the compounds on cell proliferation was detected by CellTiter-Glo luminescence assay, and the IC50 was calculated. 50 value.

[0150] Table 2. In vitro antitumor activity IC 50 (μM):

[0151] The in vitro antitumor activity IC50 of the test compound and TP 50The values ​​are shown in Table 2. All tested compounds exhibited in vitro antitumor activity, and their activity was not significantly different from that of the positive control TP, suggesting their potential for development as antitumor drugs. Among them, compounds 1, 2, 5, 10, and 11 showed in vitro antitumor activity close to that of the positive control TP, while compound 8 showed higher in vitro antitumor activity than the positive control TP.

[0152] Using triptolide and triptolide as raw materials, heteroatom substituents, heterocyclic or heteroaromatic ring substituents are introduced onto the five-membered lactone ring to obtain triptolide and triptolide derivatives. The introduction of polar groups and heteroatoms may enhance the activity of the drug molecules by forming additional hydrogen bonds or other binding interactions with amino acid residues, and may also improve the physicochemical properties of the drug molecules, thus enriching the routes of administration. Compared with triptolide (TP), some compounds obtained in this invention exhibit antitumor activity comparable to or higher than the positive control TP, showing good development and application prospects.

[0153] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound of Formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph, or prodrug thereof, in, CR3R4 is CHOH or C=O; R1 and R2 are different, and are each independently selected from the following group: C1-C8 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C6-C10 aryl, -C1-C4 alkylene-C6-C10 aryl, 3-8 membered heterocyclic or 4-8 membered heteroaryl; the above groups are optionally substituted by one or more groups selected from the following group: halogen, -OH, NR a R b CN, COOH, unsubstituted or halogenated C1-C8 alkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C1-C8 alkoxy, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogenated C2-C6 alkynyl, unsubstituted or halogenated C6-C10 aryl, unsubstituted or halogenated 4-8 membered heteroaryl, unsubstituted or halogenated 4-8 membered heterocyclic; wherein, R a R b Each is independently selected from: H, C1-C8 alkyl groups.

2. The compound according to claim 1, characterized in that, R1 and R2 are different, and are each independently selected from the following group: C1-C6 alkyl, C3-C6 cycloalkyl, C2-C4 alkenyl, C2-C4 ynyl, C6-C10 aryl, -C1-C4 alkylene-C6-C10 aryl, 5-8 membered heterocyclic or 5-8 membered heteroaryl; the above groups are optionally substituted by one or more groups selected from the following group: halogen, -OH, NR a R b CN, COOH, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C3-C6 cycloalkyl, unsubstituted or halogenated C1-C6 alkoxy, unsubstituted or halogenated C2-C4 alkenyl, unsubstituted or halogenated C2-C4 alkynyl, unsubstituted or halogenated C6-C10 aryl, unsubstituted or halogenated 5-8 membered heteroaryl, unsubstituted or halogenated 5-8 membered heterocyclic; wherein, R a R b Each is independently selected from: H, C1-C6 alkyl groups.

3. The compound according to claim 1, characterized in that, R1 is selected from the group consisting of: C6-C10 aryl, C3-C6 cycloalkyl, 5-8 membered heterocyclic, or 5-8 membered heteroaryl; the above groups are optionally substituted by one or more groups selected from the group consisting of: halogen, -OH, NR. a R b , COOH, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkoxy; wherein, R a R b Each is independently selected from: H, C1-C6 alkyl groups.

4. The compound according to claim 1, characterized in that, R2 is selected from the group consisting of: 5-8-membered heteroaryl, C1-C6 alkyl, C6-C10 aryl, and C3-C6 cycloalkyl; the above groups are optionally substituted by one or more groups selected from the group consisting of: halogen, -OH, and NR. a R b CN, COOH, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkoxy; wherein, R a R b Each is independently selected from: H, C1-C6 alkyl groups.

5. The compound according to claim 1, characterized in that, The compounds are selected from the group consisting of:

6. The method for preparing the compound according to claim 1, characterized in that, The method includes the following steps: (a) The compound of formula II reacts with the first acylation reagent to give the compound of general formula III: wherein the first acylation reagent is R1COCl, R1COBr or R1COOCOR1; (b) The compound of general formula III is reacted with a second acylation reagent to give a compound of formula I, wherein the second acylation reagent is R2COCl, R2COBr, or R2COOCOR2; In each formula, R1, R2, R3 and R4 are defined as described in claim 1.

7. The method for preparing the compound according to claim 1, characterized in that, In step (a), the reaction is carried out in an organic solvent in the presence of an organometallic base, wherein the organometallic base is selected from the group consisting of: lithium diisopropylamino (LDA), n-butyllithium (n-BuLi), tert-butyllithium (t-BuLi), lithium hexamethyldisilamide (LiHMDS), sodium hexamethyldisilamide (NaHMDS), potassium hexamethyldisilamide (KHMDS), 2,2,6,6-tetramethylpiperidinyllithium, and 2,2,6,6-tetramethylpiperidinyllithium chloride and magnesium chloride; and the organic solvent is selected from the group consisting of: tetrahydrofuran, diethyl ether, 2-methyltetrahydrofuran, or a mixture of two or more solvents; and / or In step (b), the reaction is carried out in an organic solvent in the presence of a base, wherein the base is selected from: triethylamine, diisopropylamine, N,N-diisopropylethylamine, pyridine, and piperidine; and the organic solvent is selected from the group consisting of: ethyl acetate, isopropyl acetate, dichloromethane, chloroform, tetrahydrofuran, 2-methyltetrahydrofuran, diethyl ether, acetonitrile, 1,4-dioxane, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, or a mixture of two or more solvents.

8. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises: The compound of claim 1, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph, or prodrug thereof; and Pharmaceutically acceptable carrier.

9. The use of the compound of claim 1, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph, or prodrug thereof, or the pharmaceutical composition of claim 8, characterized in that, Used to prepare drugs for treating tumors.

10. The use as described in claim 9, characterized in that, The tumors are selected from the following group: leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, squamous cell carcinoma of the lung, adenocarcinoma of the lung, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, cervical cancer, ovarian cancer, intestinal cancer, nasopharyngeal carcinoma, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, and oral cancer.

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