1, 10-split ring type guaiane sesquiterpene lactone quasi-natural product as well as preparation method and application thereof

Abstract

The invention discloses a 1, 10-split ring type guaiane sesquiterpene lactone quasi-natural product as well as a preparation method and application of the 1, 10-split ring type guaiane sesquiterpene lactone quasi-natural product. The quasi-natural product and the derivative thereof are prepared by taking (R)-Carvone as a raw material through a chemical synthesis method. Experiments show that the 1, 10-split ring type guaiane sesquiterpene lactone compound prepared by the invention has the effect of inhibiting red blood cell hemolysis, can be used as a candidate drug molecule for preventing and / or treating NLRP3 inflammasome related diseases, and has the potential of being developed into related anti-inflammatory drugs.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology and relates to ( R )-Carvone (i.e., L-carvone) was used as the starting material to prepare 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural products and their derivatives through chemical synthesis, and the anti-inflammatory activity of these compounds was tested in an established erythrocyte hemolysis model. Background Art

[0002] Inflammatory bowel disease (IBD) is a chronic, nonspecific inflammatory disease of the intestine with two clinical subtypes: ulcerative colitis (UC) and Crohn's disease (CD). IBD is more common in young and middle-aged adults. In recent years, the incidence of IBD has been increasing, making it a major threat to intestinal health after colorectal cancer. Furthermore, if the disease persists, patients may develop complications such as intestinal perforation, obstruction, intra-abdominal abscesses, and even toxic megacolon, severely impacting their quality of life and causing significant suffering.

[0003] The NLRP3 inflammasome is an organelle composed of the NLRP3 scaffold, the adaptor protein ASC, and caspase-1. Studies have shown that the NLRP3 inflammasome is closely associated with the development of several major human diseases, including inflammatory bowel disease. Although NLRP3 has gradually become an important target for drug development for inflammatory diseases, no drug has been developed since its discovery.

[0004] Sesquiterpene lactones exhibit significant anti-inflammatory potential due to their ability to target and inhibit key pro-inflammatory molecules involved in diverse inflammatory pathways and prevent or reduce inflammatory tissue damage. Although sesquiterpene lactones represent an important source for the discovery of novel anti-inflammatory compounds, only a few studies have reported on the anti-inflammatory, immunomodulatory, and anticancer effects of 1,10-split-ring guaiacolides. In fact, unmodified natural products suffer from numerous deficiencies in their pharmacological activity and drugability. For example, a 1,10-split-ring guaiacolide sesquiterpene lactone (IF-37) reported in "Chemical Constituents and Biological Activities of Four Inula Plants" was found to have low anti-inflammatory activity after experimental evaluation. The concept of pseudo-natural products (PNPs) stems from novel evolutionary strategies aimed at discovering bioactive compounds. However, there are currently no reports on the use of these compounds and their derivatives as active anti-inflammatory ingredients. Summary of the Invention

[0005] The purpose of the present invention is to provide a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product, a preparation method and use thereof.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions: In a first aspect, a 1,10-split-ring guaiacyl sesquiterpene lactone compound is provided, wherein the compound is a compound having a structure as shown in Formula I or any one of a pharmaceutically acceptable salt, tautomer, stereoisomer, and precursor compound of the compound having a structure as shown in Formula I:

[0007] In Formula I: R1 and R2 together form a double bond; or R1 is hydrogen and R2 is selected from any one of an alkylaminomethyl group, an aromatic aminomethyl group, a heteroaromatic aminomethyl group, and a group having the following structure (i.e., Formula X):

[0008] In Formula X, R3, R4, and the N atom form a cyclic structure (specifically, a heterocyclic ring containing one N atom and the rest being C atoms), and the cyclic structure is a 3- to 7-membered ring; the hydrogen atoms in the cyclic structure (which may refer to the aforementioned 3- to 7-membered ring or the ring structure of the aforementioned arylaminomethyl or heteroarylaminomethyl group) are unsubstituted or substituted with one or more groups (if one or more hydrogen atoms in the cyclic structure are substituted, the corresponding substituents may be selected from one or more of an alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, an alkylaryl group, an arylalkyl group, an arylalkenyl group, an arylalkynyl group, a heterocyclic group, a carboxyl group, a nitro group, a halogen, a carbonyl oxygen atom, a sulfur atom, and a halogen-substituted alkyl group); R5 is allyl, acetaldehyde, -C2H4ORc, -C2H4OCOORc, -C2H4OCORc, -C2H4OCONRcRd, -CH2ORc, -CH2OCOORc, -CH2OCORc or -CH2OCONRcRd; R6 is hydrogen; R7 is a methyl group; R8 is a carbonyl oxygen atom, -ORc, -OCOORc, -OCORc or -OCONRcRd; The Rc and Rd are each independently selected from any one of hydrogen, alkynyl, cycloalkyl, halogen, alkyl, aryl, halogen-substituted alkyl, halogen-substituted aryl, and nitro-substituted aryl.

[0009] In a second aspect, a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product is provided, wherein the pseudo-natural product is a compound having a structure as shown in Formula I-1, Formula I-2, Formula I-3, Formula I-4 or Formula I-5, or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers, and precursor compounds of the compound:

[0010] Among them, the 1,10-ring-split guaiacyl sesquiterpene lactone pseudo-natural product shown in formula I-1 is the compound corresponding to the case where R1 and R2 of the compound shown in formula I form a double bond, R5 is an allyl group, and R8 is a hydroxyl group; the 1,10-ring-split guaiacyl sesquiterpene lactone pseudo-natural product shown in formula I-2 is the compound corresponding to the case where R1 and R2 of the compound shown in formula I form a double bond, R5 is an acetaldehyde group, and R8 is a carbonyl oxygen atom; the 1,10-ring-split guaiacyl sesquiterpene lactone pseudo-natural product shown in formula I-3 is the compound corresponding to the case where R1 and R2 of the compound shown in formula I form a double bond, R5 is an acetaldehyde group, and R8 is a carbonyl oxygen atom. The compound shown in formula I-4 is the compound corresponding to the case where R1 and R2 form a double bond, R5 is a hydroxymethyl group and R8 is a hydroxyl group; the 1,10-ring-split guaiacyl sesquiterpene lactone pseudo-natural product shown in formula I-5 is the compound corresponding to the case where R1 and R2 form a double bond, R5 is an allyl group and R8 is a carbonyl oxygen atom in the compound shown in formula I-1.

[0011] In a third aspect, a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product derivative is provided, wherein the pseudo-natural product derivative is a compound having a structure as shown in Formula II, Formula III, Formula IV, Formula V or Formula VI (the structures of these compounds also conform to the general formula of Formula I above) or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers, and precursor compounds of the compound:

[0012] Among them, R9, R 10 Each independently selected from any one of Rc, COORc, CORc, CONRcRd, wherein Rc and Rd are each independently selected from any one of hydrogen, alkynyl, cycloalkyl, halogen, alkyl, aryl, halogen-substituted alkyl, halogen-substituted aryl, and nitro-substituted aryl (as a derivative of a compound as shown in formula I-3 or formula I-4, in a compound as shown in formula IV or formula V, R9 and R 10 are not hydrogen at the same time); R 11Any one selected from alkylamino, arylamino, heteroarylamino, and heterocyclic amino containing one N atom, wherein the heterocyclic ring containing one N atom is a 3- to 7-membered ring; the hydrogen atoms on the cyclic structure (which may refer to the aforementioned 3- to 7-membered ring or the ring structure of the aforementioned arylamino and heteroarylamino) are not substituted or are substituted with one or more groups (if one or more hydrogen atoms on the cyclic structure are substituted, the corresponding substituents may be selected from one or more of alkyl, cycloalkyl, alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, arylalkenyl, arylalkynyl, heterocyclic group, carboxyl, nitro, halogen, carbonyl oxygen atom, sulfur atom, and halogen-substituted alkyl).

[0013] Preferably, the R 11 Any one selected from dimethylamino, pyrrolidinyl, 2-carboxy-pyrrolidinyl, piperidinyl, 4-trifluoromethyl-piperidinyl, 5-nitro-indazolyl, imidazolyl, 4-nitro-imidazolyl, 1,2,4-triazolyl, 1,2,3-triazolyl, 5-fluorouracilyl, 6-mercaptopurinyl, 6-nitro-indazolyl, pyrazolyl, etc.

[0014] Preferably, Rc or Rd is selected from any one of C1-C10 (i.e., having 1-10 carbon atoms) alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-octyl, n-nonyl, etc.), C3-C7 (i.e., having 3-7 carbon atoms) cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc.), ethynyl, 2-propynyl, 3-butynyl, phenyl, halogen (e.g., chlorine, etc.), C1-C6 (i.e., having 1-6 carbon atoms) halogen-substituted alkyl (e.g., chloromethyl, bromomethyl, chloroethyl, bromoethyl, fluoroethyl, chlorobutyl, etc.), halogen-substituted phenyl (e.g., 4-fluoro-phenyl, 4-chloro-phenyl, 4-bromo-phenyl, etc.), and nitro-substituted phenyl (e.g., 4-nitro-phenyl, etc.).

[0015] Preferably, the pseudo-natural product derivative is a compound having a structure as shown in Formula II-1 to II-11, or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers, and precursor compounds of the compound:

[0016] Preferably, the pseudo-natural product derivative is a compound having a structure as shown in Formula III-1 to III-11, or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers, and precursor compounds of the compound:

[0017] Preferably, the pseudo-natural product derivative is a compound having a structure as shown in Formula VI-1 to VI-11, or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers, and precursor compounds of the compound:

[0018] Preferably, the pseudo-natural product derivative is a compound having a structure as shown in Formula IV-1 to IV-23, or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers, and precursor compounds of the compound:

[0019] Preferably, the pseudo-natural product derivative is a compound having a structure as shown in Formula V-1 to V-22, or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers, and precursor compounds of the compound:

[0020] In a fourth aspect, a method for preparing a 1,10-split-ring guaiacyl sesquiterpene lactone compound is provided, comprising the following steps: by( R )-Carvone (i.e., L-carvone) is used as a raw material, and a chemical synthesis is performed (including reacting a cyclopentane-1-carboxaldehyde intermediate with a protected hydroxyl group synthesized using L-carvone with an intermediate synthesized using propynol or butynol and retaining an alcoholic hydroxyl group and a methylene chain) to prepare a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product with a protected hydroxyl group as shown in the above formula I-3 or formula I-4 (referred to as the hydroxyl-protected compound I-3 or the hydroxyl-protected compound I-4), and the hydroxyl-protected compound I-4 is used to prepare the product (by oxidizing the corresponding alcoholic hydroxyl group contained in the structure of the hydroxyl-protected compound I-4 to an aldehyde group and reacting the generated aldehyde group to obtain a hydroxyl-protected compound I-4). The hydroxyl group is protected as a 1,10-ring-split guaiacyl sesquiterpene lactone pseudo-natural product (referred to as hydroxyl-protected compound I-1) as shown in the above formula I-1, and then the 1,10-ring-split guaiacyl sesquiterpene lactone pseudo-natural product (referred to as compound I-1, compound I-3 or compound I-4) as shown in the above formula I-1, formula I-3 and formula I-4 are prepared by deprotection reaction respectively; then, compound 1 and compound 4 are used to prepare the 1,10-ring-split guaiacyl sesquiterpene lactone pseudo-natural product (referred to as compound I-5, compound I-2) as shown in the above formula I-5 and formula I-2 respectively by oxidation reaction. Among them, the structures of hydroxyl-protected compound I-1, compound I-3 and compound I-4 are specifically shown in the following formula (in the following formula, “ PG " refers to a protecting group formed by any one of the reagents that can be used to protect the corresponding hydroxyl group, such as TBS, etc.): .

[0021] In a fifth aspect, a method for preparing a 1,10-split-ring guaiacyl sesquiterpene lactone compound is provided, comprising the following steps: by( R )-Carvone is used as the starting material, and after multiple steps of chemical transformation, compounds with structures such as those shown in Formula I-1, Formula I-2, Formula I-3, Formula I-4 or Formula I-5 are prepared respectively, and compounds with structures such as those shown in Formula II, Formula III, Formula IV, Formula V or Formula VI are prepared through reactions for achieving esterification, etherification and / or intermolecular Michael addition reactions.

[0022] Preferably, the preparation method of the 1,10-split-ring guaiacyl sesquiterpene lactone compound specifically comprises the following steps: 1) allowing the compound I-1 to undergo a condensation reaction with a carboxylic acid, anhydride, or acyl halide (e.g., acyl chloride) used to provide R9 in the presence of a condensing agent, a catalyst, and a base (or in the presence of a catalyst and a base) (in the condensation reaction, if the anhydride is highly active, the condensing agent can be omitted in the reaction) to produce an intermediate containing an acyl group or an ether bond; Alternatively, the above compound Ⅰ-1 is reacted with a halogenated hydrocarbon used to provide the R9 to undergo a nucleophilic substitution reaction to prepare an intermediate containing an ether bond; 2) Under alkaline conditions, the intermediate prepared in step 1 is reacted with the compound for providing the R 11 The organic amine undergoes a selective intermolecular Michael addition reaction to prepare the above compound II.

[0023] Preferably, the conditions of the condensation reaction include: the reaction solvent is tetrahydrofuran (THF), acetonitrile, benzene, dichloromethane (DCM), pyridine or chloroform; the condensing agent is N,N′-dicyclohexylcarbodiimide (DCC), carbonyldiimidazole (CDI), diisopropylcarbodiimide (DIC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), etc.; the catalyst is 4-dimethylaminopyridine (DMAP), 1-hydroxybenzotriazole (HOBt), etc.; the base is triethylamine, pyridine, sodium hydride, n-butyllithium or lithium diisopropylamide; the reaction temperature is 25-35°C, and the reaction time is 10-15 h; the molar ratio of compound I-1 to the carboxylic acid, anhydride or acyl halide used to provide R9 is 1:1.1-2.

[0024] Preferably, the conditions for the nucleophilic substitution reaction include: the reaction solvent is tetrahydrofuran, diethyl ether, N,N-dimethylformamide or ethylene glycol dimethyl ether; the reaction temperature is 0-85°C, the reaction time is 2-12 h; the molar ratio of compound I-1 to the halogenated hydrocarbon used to provide R9 is 1:1.1-2.2.

[0025] Preferably, the conditions for the intermolecular Michael addition reaction include: the base used (providing alkaline conditions) is selected from piperidine, triethanolamine (TEA), sodium hydroxide, potassium hydroxide, sodium ethoxide, potassium carbonate, potassium tert-butoxide or 1,8-diazabicyclo[5.4.0]undec-7-ene; the reaction solvent is methanol, tetrahydrofuran, acetonitrile, benzene, dichloromethane or chloroform; the reaction temperature is 25-35 ° C, and the reaction time is 12-20 h; the intermediate prepared in step 1 is reacted with the intermediate used to provide the R 11 The molar ratio of the organic amine is 1:1.1~2.

[0026] Preferably, the preparation method of the 1,10-split-ring guaiacyl sesquiterpene lactone compound specifically comprises the following steps: Under alkaline conditions, the above compound I-2 or compound I-5 is reacted with the compound for providing the R 11 The organic amine undergoes a selective intermolecular Michael addition reaction to prepare the above-mentioned compound III or compound VI.

[0027] Preferably, the preparation method of the 1,10-split-ring guaiacyl sesquiterpene lactone compound specifically comprises the following steps: 1) The above-mentioned hydroxy-protected compound I-3 or hydroxy-protected compound I-4 is reacted with a condensing agent, a catalyst and a base (or a catalyst and a base) to provide the R 10 (or R9) undergoes a condensation reaction with a carboxylic acid, anhydride or acyl halide (such as acyl chloride) (in the condensation reaction, if the anhydride is highly active, the condensing agent can be omitted in the reaction) to prepare an intermediate containing an acyl group or an ether bond; Alternatively, the above hydroxy protected compound I-3 or hydroxy protected compound I-4 is reacted with a 10 (or R9) undergoes a nucleophilic substitution reaction to produce an intermediate containing an ether bond; 2) subjecting the intermediate prepared in step 1 to a deprotection reaction to obtain the above-mentioned compound IV or compound V.

[0028] In a sixth aspect, an anti-inflammatory drug is provided, wherein the main active ingredient of the drug is the above-mentioned 1,10-split-ring guaiacane sesquiterpene lactone compound (see the above-mentioned formula I).

[0029] Preferably, the main active ingredient of the drug is the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product (such as Compound I-1, Compound I-3, etc.).

[0030] Preferably, the main active ingredient of the drug is the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product derivative (such as Compound II, Compound IV, etc.).

[0031] In a seventh aspect, a drug for preventing and / or treating NLRP3 inflammasome-related diseases is provided, wherein the main active ingredient of the drug is the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone compound (see Formula I above).

[0032] Preferably, the main active ingredient of the drug is the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product (such as Compound I-1, Compound I-3, etc.).

[0033] Preferably, the main active ingredient of the drug is the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product derivative (such as Compound II, Compound IV, etc.).

[0034] The above anti-inflammatory drugs and drugs for preventing and / or treating NLRP3 inflammasome-related diseases contain, in addition to the main active ingredients, a small amount of secondary ingredients and / or formulation excipients that do not affect the efficacy of the main active ingredients. For example, these drugs may contain sweeteners to improve the taste, antioxidants to prevent oxidation, and pharmaceutically acceptable carriers.

[0035] For the above-mentioned anti-inflammatory drugs and drugs for preventing and / or treating NLRP3 inflammasome-related diseases, there is no limitation on the dosage form, as long as the dosage form can effectively deliver the main active ingredients to the body. For example, these drugs can be in common dosage forms such as tablets, capsules, powders, granules, syrups, solutions, suspensions, injections, tinctures, oral liquids, aerosols, lozenges, granules, pills, powders, or sustained-release dosage forms such as nanoformulations.

[0036] In an eighth aspect, the present invention provides the use of the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone compounds (especially the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural products and their derivatives) in the preparation of anti-inflammatory drugs.

[0037] In a ninth aspect, the present invention provides the use of the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone compounds (especially the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural products and their derivatives) in the preparation of drugs for preventing and / or treating NLRP3 inflammasome-related diseases.

[0038] In a tenth aspect, the present invention provides the use of the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone compounds (especially the above-mentioned 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural products and their derivatives) in the preparation of drugs for treating nonspecific intestinal inflammatory diseases (such as inflammatory bowel disease) or arthritis.

[0039] Some of the terms appearing in the technical solutions of the present invention are defined as follows: The term "pharmaceutically acceptable salt" refers to a salt (such as sodium salt, hydrochloride, etc.) formed by the 1,10-split ring guaiacyl sesquiterpenoid lactone compound of the present invention and a base, an inorganic acid or an organic acid; the inorganic acid includes hydrofluoric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, nitric acid, sulfuric acid; the organic acid includes formic acid, acetic acid, propionic acid, succinic acid, 1,5-naphthalene disulfonic acid, asiatic acid, oxalic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, amyl Acid, diethylacetic acid, malonic acid, succinic acid, fumaric acid, pimelic acid, adipic acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methanesulfonic acid, p-toluenesulfonic acid, citric acid, oleic acid, glycolic acid, mercaptan, sarcosine, glycine and other amino acids; "pharmaceutically acceptable" means suitable for use in humans without excessive adverse side effects (such as toxicity, irritation and allergic reactions), that is, with a reasonable benefit / risk ratio.

[0040] The term "tautomer" refers to functional isomers that result from the rapid shift of an atom between two positions in a molecule.

[0041] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, such as cis-trans isomers, enantiomers, conformers, and the like.

[0042] The term "precursor compound" refers to a compound that is inactive in vitro but can be converted into a pharmaceutically active compound through metabolism or chemical reactions in the body.

[0043] The beneficial effects of the present invention are embodied in: The present invention conducts drug activity tests and pharmacological analyses on the synthesized 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural products and their derivatives having the general formula I. The results show that the compounds exhibit an inhibitory effect on erythrocyte hemolysis in an experiment to inhibit erythrocyte hemolysis activity, especially compounds I-1, I-3, II-1, III-9, IV-2 and VI-4, which have an inhibitory effect superior to (or equivalent to) that of the positive control drug MCC950 that specifically targets the NLRP3 inflammasome at lower concentrations. The present invention screens candidate drug molecules for NLRP3 inflammasome-related diseases through cell-level experiments, suggesting that 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural products and their derivatives with the general formula I structure can inhibit inflammation and have the potential and medicinal value to be developed as anti-inflammatory drugs for the prevention and / or treatment of NLRP3-related inflammatory diseases such as inflammatory bowel disease. At the same time, it provides a parent or core structure (such as compound I-1, compound I-3, etc.) for subsequent large-scale drug molecule synthesis and screening, which is of great significance for broadening the application range of 1,10-split-ring guaiacyl lactone compounds. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1-1 This is the synthetic route of compound 2, compound 6a and compound 6b.

[0045] Figure 1-2 This is the synthetic route for compound 8a, compound 8b and compound 17.

[0046] Figure 1-3 The invention relates to a synthetic route for 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural products and their derivatives using the general formula I structure.

[0047] Figure 2 This is the synthetic route of compound Ⅰ-3.

[0048] Figure 3 This is the synthetic route of compound Ⅰ-4.

[0049] Figure 4 This is the synthetic route of compound Ⅰ-2.

[0050] Figure 5 This is the synthetic route of compound Ⅰ-1.

[0051] Figure 6 This is the synthetic route of compound Ⅰ-5.

[0052] Figure 7 This is the synthetic route of compound III-9.

[0053] Figure 8 This is the synthetic route of compound II-1.

[0054] Figure 9 This is the synthetic route of compound II-1-HCl.

[0055] Figure 10 This is the synthetic route of compound IV-2.

[0056] Figure 11 This is the synthetic route of compound V-22.

[0057] Figure 12 This is the synthetic route of compound VI-4.

[0058] Figure 13 The experimental results of representative compounds Ⅰ-1, Ⅰ-2, Ⅰ-3, Ⅰ-4, Ⅰ-5, Ⅱ-1, Ⅲ-9, Ⅳ-2, Ⅴ-22 and Ⅵ-4 inhibiting erythrocyte hemolysis activity (OD505 test data after administration). DETAILED DESCRIPTION

[0059] The present invention will be further described below with reference to the accompanying drawings and examples. It should be understood that the examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0060] (I) Design and synthesis of 1,10-split-ring guaiacyl sesquiterpenoid lactone pseudo-natural products and their derivatives See also Figure 1-1 、 Figure 1-2 and Figure 1-3 The present invention uses (R)-Carvone (i.e., compound 1), propargyl alcohol (i.e., compound 3a), and butynol (i.e., compound 3b) as raw materials, respectively, and adopts a designed series of synthetic routes to first prepare important intermediates (i.e., compound 2, compound 6a, and compound 6b; wherein compound 6a and compound 6b are obtained by firstly subjecting propargyl alcohol and butynol, respectively, after protecting their alcoholic hydroxyl groups, to a nucleophilic substitution reaction in the presence of n-butyl lithium, and then reacting them with diisobutylaluminum hydride to an electrophilic substitution reaction with iodoborane pinacol ester). Then, these important intermediates are used (specifically, compound 2 and compound 6a, and compound 2 and compound 6b, respectively, are subjected to a tandem lactonization reaction in the presence of boron trifluoride etherate), and the corresponding alcoholic hydroxyl protecting groups from compounds 6a and 6b are removed, thereby Compounds 8a, 8b, and 17 were prepared; compound 17 was obtained by reacting compound 8b in the presence of a Dess-Martin periodinane and then subjecting the reaction product to a Wittig reaction. A novel 1,10-ring-split guaiacyl sesquiterpene lactone pseudo-natural product (including compounds I-1 and I-5 prepared using compound 17, compound I-3 prepared using compound 8a, and compounds I-4 and I-2 prepared using compound 8b) and derivatives of the 1,10-ring-split guaiacyl sesquiterpene lactone pseudo-natural product (including compound II prepared using compound I-1, compound III prepared using compound I-2, compound IV prepared using compound 8a, compound V prepared using compound 8b, and compound VI prepared using compound I-5) were prepared. The syntheses of compounds 8a, 8b, and 17 are described in detail below.

[0061] 1.1 Synthesis of compound 2 According to the method reported in the literature (reference: Elford TG, Hall D G. Total synthesis of (+)-chinensiolide B via tandem allylboration / lactonization. Journal of the American Chemical Society. 2010, 132(5):1488-1489.), (R)-Carvone (i.e., compound 1) was used as the raw material and cyclopentane-1-carboxaldehyde compounds (specifically compound 2) were prepared through 6 steps of transformation. 1H NMR (400MHz, CDCl3) δ 9.52 (d, J = 3.8 Hz, 1H), 4.88 – 4.66 (m, 2H), 4.18 (t, J = 3.7 Hz,1H), 3.60 (s, 1H), 3.30 (t, J = 10.9 Hz, 1H), 2.61 (m, 1H), 2.34 – 2.20 (m,1H), 1.92 – 1.78 (m, 1H), 1.72 (d, J = 1.5 Hz, 3H), 0.98 (d, J = 6.8 Hz, 3H),0.89 (d, J = 2.3 Hz, 9H), 0.05 (d, J = 4.8 Hz, 6H). The NMR identification results of compound 2 were consistent with those reported in the literature.

[0062] 1.2 Synthesis of compound 6a (1) To a 250 mL reaction flask containing a magnetic stirrer, 110 mL of THF, 5 g of propargyl alcohol (i.e., compound 3a; 89.18 mmol, 1.0 equiv), and 15.49 g of imidazole (222.96 mmol, 2.5 equiv) were added. Then, 28.32 mL of tert-butyldiphenylsilyl chloride (TBDPSCl; 107.02 mmol, 1.2 equiv) was slowly added dropwise to the reaction flask through a syringe. The addition time was about 25 minutes, and stirring was continued at room temperature for 1.5 hours. After TLC monitoring confirmed that the raw materials had reacted completely, 30 mL of saturated NH4Cl solution was added to terminate the reaction. 30 mL of water was added, and the mixture was extracted with ethyl acetate (3×30 mL). The organic phases were combined and washed with 50 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using PE:EA = 50:1 as the eluent to obtain 21.96 g of a colorless oil (i.e., compound 4a); 1 H NMR (400 MHz, CDCl3) δ 7.72 (d, J = 7.8 Hz,4H), 7.48 – 7.35 (m, 6H), 4.33 (d, J = 2.4 Hz, 2H), 2.38 (t, J = 2.4 Hz, 1H),1.08 (s, 9H).

[0063] (2) Under nitrogen (N2) atmosphere and -78°C, 254 mL of anhydrous THF and 15 g of compound 4a (50.94 mmol, 1.0 equiv.) were added to a 1 L round-bottom flask containing a magnetic stirrer. 35.02 mL of 1.6 M n-butyllithium hexane solution (containing 56.03 mmol, 1.1 equiv. of n-butyllithium) was slowly added dropwise to the round-bottom flask for about 20 minutes. The temperature was then maintained constant and stirring was continued for 1 hour. Subsequently, 11.93 mL of methyl chloroformate (152.82 mmol, 3 equiv.) was slowly added dropwise to the round-bottom flask at -78°C through a syringe. Stirring was continued for 1.0 hour. The round-bottom flask was then slowly warmed to room temperature. After TLC monitoring confirmed that the reaction of the raw materials was complete, 30 mL of saturated NaHCO3 solution was added to terminate the reaction. The reaction was then quenched with ethyl acetate (3×40 The combined organic phases were washed with 80 mL of saturated brine, dried over anhydrous NaSO, filtered, and the filtrate concentrated under reduced pressure to yield a crude product. The crude product was purified by silica gel column chromatography using a 40:1 PE:EA ratio to afford 7.64 g of a colorless oil (compound 5a). 1 H NMR (400 MHz, CDCl3) δ 7.67 (d, J = 6.6 Hz, 4H), 7.46 – 7.37 (m, 6H), 3.83 (d, J = 7.1 Hz, 2H),3.76 (s, 3H), 2.58 (t, J = 6.8 Hz, 2H), 1.06 (s, 9H).

[0064] (3) Under nitrogen atmosphere and 0°C, 87 mL of toluene and 7.62 mL of hexamethylphosphoric triamide (HMPA; 43.35 mmol, 2.0 equiv.) were added to a 250 mL round-bottom flask containing a magnetic stirrer. The mixture was stirred for 20 min. Then, 21.67 mL of 1 M diisobutylaluminum hydride (DIBAL-H) ​​n-hexane solution (containing 32.51 mmol, 1.5 equiv. of DIBAL-H) ​​was slowly added dropwise to the round-bottom flask. The addition time was about 15 min. Then, 7.64 g of compound 5a (21.67 mmol, 1.0 equiv.) was added to the round-bottom flask at one time. The mixture was stirred for 5 h. Then, 8.06 mL of iodomethylpinacol borate (43.35 mmol, 2.0 equiv.) was added to the round-bottom flask through a syringe and stirred for 30 min. The mixture was stirred at room temperature overnight and monitored by TLC. After confirming complete reaction, 20 mL of 1 M HCl solution was added to the round-bottom flask to terminate the reaction. 40 mL of water was then added, followed by extraction with ethyl acetate (3 × 40 mL). The organic phases were combined and washed with 30 mL of 1 M HCl solution, 50 mL of saturated NaHCO₃ solution, and 50 mL of water. The organic phases were then dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a PE:EA ratio of 40:1 as the eluent to obtain 7.28 g of a yellow oil (i.e., compound 6a). 1 H NMR (400 MHz, CDCl3) δ 7.65 (m, 4H), 7.39 –7.30 (m, 6H), 6.22 (t, J = 4.7 Hz, 1H), 4.67 (dd, J = 4.8, 1.5 Hz, 2H), 3.56 (s,3H), 1.26 (s, 2H), 1.21 (s, 6H), 1.03 (d, J = 2.0 Hz, 9H).

[0065] 1.3 Synthesis of compound 6b (1) The difference from 1.2 is that 90 mL of THF, 5 g of butynol (i.e., compound 3b; 71.34 mmol, 1.0 equiv), and 12.39 g of imidazole (178.34 mmol, 2.5 equiv) were added to a 250 mL reaction flask containing a magnetic stirrer. Then, 22.71 mL of TBDPSCl (85.6 mmol, 1.2 equiv) was slowly added dropwise to the reaction flask via a syringe. Finally, 21 g of a colorless oil (i.e., compound 4b) was obtained.1 H NMR (400 MHz, CDCl3) δ 7.75 – 7.58 (m, 4H),7.47 – 7.33 (m, 6H), 3.80 (t, J = 7.1 Hz, 2H), 2.45 (t, J = 7.1 Hz, 2H), 1.94 (t, J = 2.6 Hz, 1H), 1.07 (s, 9H).

[0066] (2) The difference from 1.2 is that: under nitrogen atmosphere and at -78 °C, 162 mL of anhydrous THF and 15 g of compound 4b (48.62 mmol, 1.0 equiv.) were added to a 1 L round-bottom flask with a magnetic stirrer. Then, 33.43 mL of 1.6 M n-butyllithium hexane solution (containing 53.49 mmol, 1.1 equiv. of n-butyllithium) was slowly added dropwise to the round-bottom flask. The addition time was about 20 minutes. After that, the temperature was kept constant and stirring was continued for 1 hour. Then, 11.38 mL of methyl chloroformate (145.87 mmol, 3 equiv.) was slowly added dropwise to the round-bottom flask at -78 °C through a syringe. Finally, 12.64 g of a colorless oil (i.e., compound 5b) was obtained. 1 H NMR (400 MHz, CDCl3) δ 7.69 (d, J = 6.5 Hz, 4H), 7.51 – 7.33(m, 6H), 4.41 (s, 2H), 3.78 (s, 3H), 1.07 (s, 9H).

[0067] (3) The difference from 1.2 is that: under nitrogen atmosphere and 0°C, 87 mL of toluene and 7.67 mL of hexamethylphosphoric triamide (HMPA; 43.65 mmol, 2.0 equiv.) were added to a 250 mL round-bottom flask with a magnetic stirrer. The mixture was stirred for 20 min, and then 21.83 mL of 1 M diisobutylaluminum hydride (DIBAL-H) ​​n-hexane solution (containing 32.74 mmol, 1.5 equiv. of DIBAL-H) ​​was slowly added dropwise to the round-bottom flask. The addition time was about 15 min. Then, 8 g of compound 5b (21.83 mmol, 1.0 equiv.) was added to the round-bottom flask at one time. Stirring was continued for 5 h, and then 8.12 mL of iodomethylpinacol borate (43.65 mmol, 2.0 equiv.) was added to the round-bottom flask through a syringe. Finally, 6.42 g of yellow oil (i.e., compound 6b) was obtained; 1H NMR (400 MHz, CDCl3) δ 7.71 – 7.63 (m, 4H), 7.44 – 7.34(m, 6H), 6.03 (t, J = 7.2 Hz, 1H), 3.74 (d, J = 6.5 Hz, 2H), 3.70 (s, 3H), 2.78(q, J = 6.8 Hz, 1H), 1.85 (s, 2H), 1.28 (s, 2H), 1.23 (s, 12H), 1.05 (S, 9H).

[0068] 1.4 Synthesis of compound 8a (1) Under nitrogen atmosphere and 0°C, 37 mL of anhydrous toluene, 1.05 g of compound 2 (3.72 mmol, 1.0 equiv.), and 3.68 g of compound 6a (7.43 mmol, 2.0 equiv.) were added to a 100 mL reaction flask containing a magnetic stirrer. 47.85 µL of boron trifluoride etherate (371.68 µmol, 0.1 equiv.) was then added to the reaction flask via a microsyringe. The mixture was stirred at 0°C for 6 h, then warmed to room temperature and stirred for 48 h. After TLC monitoring confirmed that the reaction of the raw materials was complete, 15 mL of a mixed solution of NH4Cl:NH4OH = 9:1 was added to terminate the reaction. 20 mL of water was added, and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined and washed with 20 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using PE:EA=50:1 as eluent to obtain 1.51 g of a light yellow oil (i.e., compound 7a); 1 H NMR (400 MHz, CDCl3) δ 7.68 –7.57 (m, 4H), 7.48 – 7.43 (m, 2H), 7.42 – 7.36 (m, 4H), 6.23 (d, J = 2.6 Hz,1H), 5.51 (d, J = 2.2 Hz, 1H), 4.90 (s, 1H), 4.84 (s, 1H), 4.48 (m, 1H), 4.14(t, J= 4.5 Hz, 1H), 3.69 (m, 2H), 3.05 (m, 1H), 2.93 (s, 1H), 2.06 – 1.99 (m,2H), 1.97 (s, 1H), 1.77 (s, 3H), 1.61 (s, 1H), 1.04 (d, J = 6.1 Hz, 9H), 1.01(s, 3H), 0.89 (s, 9H), 0.03 (s, 6H). IR (neat): 3071, 2954, 2928, 2856, 1763,1651 cm -1 ; 13 C NMR (151 MHz, CDCl3) δ 170.58, 144.27, 136.60, 135.58, 132.81,132.79, 129.98, 127.86, 127.83, 122.63, 112.55, 81.14, 77.25, 77.04, 76.83,74.62, 65.39, 50.81, 45.99, 45.36, 39.87, 38.44, 26.75, 25.90, 23.39, 19.17,18.19, 17.12, -4.64, -4.99.

[0069] (2) Under nitrogen atmosphere, 22 mL of anhydrous THF and 1.35 g of compound 7a (2.18 mol, 1.0 equiv.) were added to a 50 mL reaction flask containing a magnetic stirrer. The mixture was cooled to 0 °C in an ice-water bath, and 2.84 mL of TBAF (2.84 mmol, 1.3 equiv.) was added to the reaction flask. The mixture was then transferred to room temperature and stirred for 2 h. After TLC monitoring confirmed that the reaction of the raw materials was complete, 30 mL of water was added to terminate the reaction. The mixture was then extracted with ethyl acetate (3×20 mL). The organic phases were combined and washed with 20 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography using PE:EA=5:1 as the eluent to obtain 672.3 mg of a colorless oil (i.e., compound 8a). 1 H NMR (400 MHz, CDCl3) δ 6.27 (d, J = 2.8 Hz, 1H), 5.66 (d, J = 2.4 Hz,1H), 4.90 (t, J= 1.8 Hz, 1H), 4.84 – 4.81 (m, 1H), 4.35 (d, J = 5.2 Hz, 1H),4.14 – 4.11 (m, 1H), 3.70 (d, J = 6.3 Hz, 2H), 3.02 (m, J = 1H), 2.97 – 2.90 (m,1H), 1.99 (m, 3H), 1.77 (d, J = 1.2 Hz, 3H), 1.61 – 1.56 (m, 1H), 1.23 (s, 3H), 1.02 (s, 2H), 0.85 (s, 9H), 0.00 (s, 6H); 13 C NMR (101 MHz, CDCl3) δ 170.45,144.33, 136.38, 122.81, 112.40, 80.90, 74.52, 63.85, 50.59, 45.93, 45.30,39.83, 38.55, 29.69, 25.87, 23.40, 18.16, 17.06, -4.66, -5.01.

[0070] 1.5 Synthesis of compound 8b (1) The difference from 1.4 is that under nitrogen atmosphere and at 0°C, 35 mL of anhydrous toluene, 1 g of compound 2 (3.54 mmol, 1.0 equiv.), and 3.6 g of compound 6b (7.08 mmol, 2.0 equiv.) were added to a 100 mL reaction bottle containing a magnetic stirrer. Then, 45.57 μL of boron trifluoride etherate (353.98 μmol, 0.1 equiv.) was added to the reaction bottle via a microsyringe. Finally, 991.1 mg of a light yellow oil (i.e., compound 7b) was obtained. 1 H NMR (400 MHz, CDCl3) δ 7.69 – 7.59 (m, 6H), 7.45 – 7.39 (m, 6H), 6.18 (d, J = 2.6 Hz, 1H),5.47 (d, J = 2.2 Hz, 1H), 4.88 (s, 1H), 4.81 (s, 1H), 4.13 (d, J= 1.8 Hz, 2H),3.79 – 3.61 (m, 2H), 3.07 – 2.88 (m, 2H), 1.98 (m, 2H), 1.94 – 1.84 (m, 1H),1.81 – 1.73 (m, 2H), 1.70 (s, 3H), 1.59 (m, 1H), 1.07 (s, 9H), 1.01 (d, J = 7.0Hz, 3H), 0.89 (s, 9H), 0.02 (s, 6H); 13 C NMR (151 MHz, CDCl3) δ 170.33, 143.65,138.71, 135.09, 132.85, 129.41, 127.35, 121.46, 112.06, 82.95, 76.80, 76.59,76.38, 74.10, 59.93, 52.98, 49.76, 45.46, 39.36, 38.88, 37.92, 36.13, 26.44,25.44, 22.98, 18.69, 17.72, 16.62, -5.11, -5.45.

[0071] (2) The difference from 1.4 is that: Under nitrogen atmosphere, 24 mL of anhydrous THF and 1.5 g of compound 7b (2.37 mol, 1.0 equiv.) were added to a 50 mL reaction flask containing a magnetic stirrer. The flask was cooled to 0 °C in an ice-water bath, and then 3.08 mL of TBAF (3.08 mmol, 1.3 equiv.) was added to the reaction flask. Finally, 552.8 mg of a colorless oil (i.e., compound 8b) was obtained. 1 H NMR (600 MHz, CDCl3) δ 6.22 (d, J = 2.8 Hz, 1H), 5.58 (d, J = 2.5 Hz, 1H),4.90 (t, J = 1.7 Hz, 1H), 4.86 – 4.79 (m, 1H), 4.21 (d, J = 5.5 Hz, 1H), 4.11 (t, J = 4.2 Hz, 1H), 3.80 – 3.66 (m, 2H), 3.05 – 2.98 (m, 1H), 2.94 (t, J= 7.5Hz,1H), 2.02 – 1.92 (m, 3H), 1.84 – 1.72 (m, 6H), 1.57 (m, J = 1H), 1.03 (s 6.7Hz, 3H), 0.85 (s, 9H), -0.00 (s, 6H); 13 C NMR (151 MHz, CDCl3) δ 170.80,144.22, 139.29, 121.86, 112.45, 83.71, 77.05, 76.84, 74.54, 59.49, 50.02,45.93, 39.89, 39.71, 38.51, 36.23, 23.49, 18.17, 17.05, -4.66, -5.00.

[0072] 1.6 Synthesis of compound 17 (1) Under nitrogen atmosphere and 0°C, 20 mL of anhydrous CH2Cl2 and 330.2 mg of compound 8b (0.84 mmol, 1.00 equiv.) were added to a 50 mL round-bottom flask containing a magnetic stirrer. 485.7 mg of Dess-Martin oxidant (1.14 mmol, 1.30 equiv.) was then added to the round-bottom flask under stirring. The temperature was raised to room temperature and stirring was continued for 1 h. After TLC monitoring confirmed that the reaction of the raw materials was complete, 15 mL of saturated Na2S2O3 solution was added to quench the reaction. The reaction was then extracted with dichloromethane (3×15 mL). The organic phases were combined and washed with 20 mL of saturated NaHCO3 solution and 20 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using PE:EA=5:1 as the eluent to obtain 239 mg of a white solid (i.e., compound 16). 1 H NMR (600 MHz, CDCl3) δ 9.79 (s, 1H), 6.23 (d, J = 2.8 Hz, 1H), 5.56 (d, J = 2.4 Hz, 1H), 4.88 (t, J = 1.8 Hz, 1H), 4.81 (s, 1H), 4.12 (t, J = 4.9 Hz, 1H), 4.10 (m, 1H), 3.29 (d, J= 6.7 Hz, 1H), 3.02 (m, 1H), 2.78 – 2.70 (m, 2H), 2.08(m, 1H), 1.99 – 1.93 (m, 2H), 1.72 (s, 3H), 1.59 – 1.57 (m, 1H), 1.02 (d, J =7.0 Hz, 3H), 0.85 (s, 9H), 0.00 (s, 6H); 13 C NMR (151 MHz, CDCl3) δ 198.89,169.97, 144.19, 138.34, 122.72, 112.40, 83.11, 77.25, 77.04, 76.83, 74.50,49.71, 48.37, 45.83, 39.79, 38.40, 37.07, 23.46, 18.17, 17.04, -4.67, -5.00.

[0073] (2) Under nitrogen atmosphere, 25 mL of THF and 1.33 g of methyltriphenylphosphonium bromide (3.65 mmol, 6.0 equiv.) were added to a 50 mL reaction flask containing a magnetic stirrer. The flask was cooled to -50 °C, and 1.5 mL of 2.5 M n-butyllithium hexane solution (containing 3.65 mmol, 6.0 equiv. of n-butyllithium) was added dropwise to the reaction flask under stirring. The stirring was continued at -50 °C for 1 h. Subsequently, 239 mg of compound 16 (0.61 mmol, 1.0 equiv.) dissolved in 5.0 mL of anhydrous THF was added dropwise to the reaction flask over about 8 min. The mixture was stirred at -50 °C for 30 min and then transferred to room temperature and stirred for 3 h. After the reaction was confirmed to be complete by TLC monitoring, 10 mL of saturated NH4Cl solution was added to quench the reaction. The mixture was then extracted with ethyl acetate (3 × 10 mL). The organic phases were combined and precipitated with 20 The product was washed with saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was spin-dried to obtain a crude product. The crude product was purified by silica gel column chromatography using a PE:EA ratio of 10:1 to obtain 153.1 mg of a colorless oil (Compound 17). 1 H NMR (600 MHz, CDCl3) δ 6.22 (d, J =2.9 Hz, 1H), 5.72 (m, 1H), 5.57 (d, J = 2.5 Hz, 1H), 5.16 – 5.05 (m, 2H), 4.89(t, J= 1.8 Hz, 1H), 4.82 (s, 1H), 4.16 (d, J = 6.1 Hz, 1H), 4.12 (t, J = 4.8 Hz,1H), 2.99 (s, 1H), 2.85 – 2.74 (m, 1H), 2.41 – 2.32 (m, 1H), 2.31 – 2.21 (m,1H), 2.04 – 1.96 (m, 2H), 1.93 (m, 1H), 1.74 (s, 3H), 1.56 (q, J = 5.7 Hz, 2H),1.01 (d, J = 7.0 Hz, 3H), 0.85 (s, 9H), 0.00 (s, 6H); 13 C NMR (151 MHz, CDCl3) δ170.65, 144.07, 138.73, 133.99, 121.78, 118.38, 112.32, 82.86, 77.25, 77.03,76.82, 74.49, 49.58, 45.88, 41.98, 39.81, 38.24, 37.54, 25.88, 23.49, 18.18,17.19, -4.65, -5.01.

[0074] (II) Synthesis examples of 1,10-ring-split guaiacyl sesquiterpenoid lactone pseudo-natural products and their derivatives 2.1 Preparation of Compound I-3 See also Figure 2To a 20 mL reaction flask containing a magnetic stirrer under a nitrogen atmosphere at 0°C, 5.7 mL of anhydrous THF and 75.7 mg of compound 8a (0.199 mmol, 1.0 equiv.) were added (slowly dropwise) to the reaction flask. The mixture was then transferred to room temperature and stirred under a nitrogen atmosphere for 2 h. After TLC confirmed the complete reaction of the starting materials, 30 mL of water was added to terminate the reaction. The reaction was then extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, washed with 20 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure (25–35°C, 0.08–0.09 MPa) to obtain the crude product. The crude product was purified by silica gel column chromatography using a PE:EA ratio of 1:1. The collected eluate was dried under vacuum to obtain 27.5 mg of a white solid, Compound I-3. The specific synthesis results and identification of Compound I-3 are as follows.

[0075] Yield 52%. f = 0.3 (PE: EA = 1:1). HPLC purity (99%). mp: 148.7 ~ 153.8℃. 1 H NMR (600 MHz, CDCl3) δ 6.27 (d, J = 2.7 Hz, 1H), 5.67 (d, J = 2.3 Hz, 1H), 4.91 (s, 1H), 4.84 (s, 1H), 4.36 (d, J = 5.1 Hz, 1H), 4.20 (t, J = 4.8Hz, 1H), 3.71 (d, J = 6.3 Hz, 2H), 3.04 (dd, J = 13.4, 7.1 Hz, 1H), 2.93 (dd,J = 5.8, 2.9 Hz, 1H), 2.11 – 1.99 (m, 3H), 1.77 (s, 3H), 1.69 (dd, J = 13.3,6.2 Hz, 1H), 1.10 (d, J = 6.5 Hz, 3H). 13C NMR (151 MHz, CDCl3) δ 170.41,143.91, 136.22, 123.10, 112.78, 80.85, 77.26, 77.05, 76.84, 74.28, 63.85,50.59, 46.05, 45.35, 39.13, 38.17, 23.25, 16.09. IR (neat): 3394, 2929, 1742,1648 cm -1 HRMS(ESI) m / z: [M+H] + Calculated for (based on the molecular formula below) C 15 H 22 O4267.1590; found (actual measured value) 267.1592.

[0076] 2.2 Preparation of Compound I-4 See also Figure 3 To a 25-mL reaction vial containing a magnetic stirrer under nitrogen at 0°C, 6 mL of anhydrous THF and 110.1 mg of compound 8b (279 µmol, 1.00 equiv.) were added (via syringe) to the reaction vial. The mixture was then transferred to room temperature and stirred under nitrogen for 2 h. After TLC confirmed the complete reaction of the starting material, 10 mL of water was added to quench the reaction. The mixture was then extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, washed with 20 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure (25–35°C, 0.08–0.09 MPa) to obtain the crude product. The crude product was purified by silica gel column chromatography using a PE:EA ratio of 1:1. The collected eluate was dried under vacuum to obtain 38.8 mg of a colorless oil, Compound I-4. The detailed synthesis results and identification of Compound I-4 are as follows.

[0077] Yield 50%. HPLC purity (97%). f = 0.2 (PE: EA = 1:1). 1H NMR (600 MHz, CDCl3) δ 6.23 (d, J = 2.8 Hz, 1H), 5.59 (d, J = 2.4 Hz, 1H), 4.91 (s, 1H), 4.84 (s, 1H), 4.26 – 4.15 (m, 2H), 3.72 (t, J = 6.2 Hz, 2H), 3.05 (m, 1H), 2.95 (m, 1H), 2.11 – 1.99 (m, 3H), 1.89 – 1.80 (m, 1H), 1.76 (s, 3), 1.69(dd, J = 13.1, 6.3 Hz, 1H), 1.49 (s, 1H), 1.10 (d, J = 6.9 Hz, 3H). 13 C NMR(151 MHz, CDCl3) δ 170.69, 143.80, 139.10, 122.10, 112.85, 83.57, 77.26,77.05, 76.84, 74.34, 59.40, 50.01, 46.07, 39.19, 38.17, 36.13, 23.29, 16.03. IR (neat): 3401, 2916, 1748, 1647 cm -1 HRMS(ESI) m / z: [M+H] + calculated for C 16 H 24 O4281.1747; found 281.1759.

[0078] 2.3 Preparation of Compound Ⅰ-2 See also Figure 4Under nitrogen atmosphere and 0 ° C, 2 mL of anhydrous CH2Cl2 and 53.2 mg of compound I-4 (189.75 μmol, 1.00 equiv.) were added to a 20 mL reaction bottle containing a magnetic stirrer. Then, 120.7 mg of Dess-Martin periodinane (0.28 mmol, 1.5 equiv.) was added to the reaction bottle under stirring. The temperature was then raised to room temperature and stirred under nitrogen atmosphere for 2 h. After TLC monitoring confirmed that the raw material reaction was complete, 5 mL of saturated Na2S2O3 solution was added to terminate the reaction. The reaction was then extracted with DCM (3×10 mL). The organic phases were combined and washed with 10 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure (25-35 ° C, 0.08-0.09 MPa) to obtain the crude product. The crude product was purified by silica gel column chromatography using a PE:EA ratio of 2:1. The collected eluate was dried under vacuum to obtain 34.4 mg of a colorless oil, Compound I-2. The detailed synthesis results and identification of Compound I-2 are as follows.

[0079] Yield 66%. HPLC purity (99%). f = 0.6 (PE: EA = 2:1). 1 H NMR (600 MHz, CDCl3) δ 9.80 (s, 1H), 6.28 (d, J = 2.8 Hz, 1H), 5.63 (d, J = 2.4 Hz, 1H), 4.98(q, J = 1.4 Hz, 1H), 4.95 – 4.80 (m, 1H), 4.17 (t, J = 4.8 Hz, 1H), 3.26 (t, J =7.5Hz, 1H), 3.06 (t, J = 11.2 Hz, 1H), 2.91 – 2.75 (m, 2H), 2.69 (m, 1H), 2.46(t, J = 7.0 Hz, 1H), 2.34 – 2.17 (m, 2H), 1.74 (s, 3H), 1.22 (d, J = 7.6 Hz, 3H). 13C NMR (151 MHz, CDCl3) δ 218.46, 198.86, 198.85, 168.88, 142.29, 137.30,123.88, 113.51, 82.67, 77.26, 77.05, 76.83, 49.40, 48.85, 43.46, 43.17, 41.07, 36.89, 21.98, 17.29. IR (neat): 2928, 1763, 1739, 1650 cm -1 HRMS(ESI) m / z: [M+H] + calculated for C 16 H 20 O4277.1434; found 277.1445.

[0080] 2.4 Preparation of Compound Ⅰ-1 See also Figure 5 Under nitrogen atmosphere and 0°C, 4 mL of anhydrous THF and 64.6 mg of compound 17 (165.37 µmol, 1.00 equiv.) were added to a 20 mL reaction flask containing a magnetic stirrer. 0.25 mL of TBAF (248.06 µmol, 1.5 equiv.) was then added to the reaction flask. The mixture was then transferred to room temperature and stirred under nitrogen atmosphere for 2 h. After TLC monitoring confirmed that the reaction of the raw materials was complete, 10 mL of water was added to quench the reaction. The mixture was then extracted with ethyl acetate (3×10 mL). The organic phases were combined and washed with 20 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure (25-35°C, 0.08-0.09 MPa) to obtain the crude product. The crude product was purified by silica gel column chromatography using a PE:EA ratio of 2:1. The collected eluate was dried under vacuum to obtain 16.8 mg of a colorless oil, Compound I-1. The detailed synthesis results and identification of Compound I-1 are as follows.

[0081] Yield 53%. HPLC purity (99%). f = 0.25 (PE:EA = 3:1). 1 H NMR (600 MHz, CDCl3) δ 6.23 (d, J = 2.9 Hz, 1H), 5.71 (m, 1H), 5.58 (d, J= 2.6 Hz, 1H), 5.15 –5.05 (m, 2H), 4.90 (s, 1H), 4.83 (s, 1H), 4.20 (t, J = 5.0 Hz, 1H), 4.17 (d, J =6.0 Hz, 1H), 3.03 (m, 1H), 2.80 (t, J = 6.3 Hz, 1H), 2.35 (m, 1H), 2.12 – 2.03(m, 2H), 1.98 (m, 1H), 1.73 (s, 3H), 1.68 (d, J = 13.2 Hz, 1H), 1.10 (d, J = 7.2 Hz, 3H). 13 C NMR (151 MHz, CDCl3) δ 170.55, 143.59, 138.53, 133.89, 122.04,118.48, 112.72, 82.75, 77.25, 77.04, 76.83, 74.25, 49.60, 46.01, 42.10,39.15, 37.87, 37.53, 23.32, 16.19. IR (neat): 3416, 2924, 1750, 1641 cm -1 HRMS(ESI) m / z: [M+H] + calculated for C 17 H 24 O3277.1798; found 277.1802.

[0082] 2.5 Preparation of Compound Ⅰ-5 See also Figure 6Under nitrogen atmosphere and 0°C, 2 mL of anhydrous CH2Cl2 and 24.4 mg of compound Ⅰ-1 (88.29 μmol, 1.00 equiv.) were added to a 20 mL reaction bottle containing a magnetic stirrer. 51.2 mg of Dess-Martin periodinane (0.12 mmol, 1.30 equiv.) was then added to the reaction bottle under stirring. The mixture was then warmed to room temperature and reacted under nitrogen atmosphere for 2 h. After TLC monitoring confirmed that the reaction of the raw materials was complete, 10 mL of saturated Na2S2O3 solution was added to terminate the reaction. The mixture was then extracted with DCM (3×10 mL). The organic phases were combined and washed with 10 mL of saturated NaHCO3 solution and saturated brine, respectively, and then dried over anhydrous Na2SO4. The filtrate was concentrated under reduced pressure (25-35°C, 0.08-0.09 MPa) to obtain the crude product. The crude product was purified by silica gel column chromatography using a PE:EA ratio of 2:1. The collected eluate was dried under vacuum to obtain 22.9 mg of a colorless oil, Compound I-5. The specific synthesis results and compound identification of Compound I-5 are as follows.

[0083] Yield 94%. HPLC purity (97%). f = 0.5 (PE:EA = 2:1). 1 H NMR (600 MHz, CDCl3) δ 6.27 (d, J = 2.8 Hz, 1H), 5.71 (m, 1H), 5.63 (d, J = 2.5 Hz, 1H), 5.14(d, J = 5.6 Hz, 1H), 5.14 – 5.11 (m, 1H), 5.00 (q, J = 1.3 Hz, 1H), 4.91 – 4.89(m, 1H), 4.25 (t, J = 5.6 Hz, 1H), 3.02 (dd, J = 11.2, 7.6 Hz, 1H), 2.79 – 2.74(m, 1H), 2.71 (m, 1H), 2.42 – 2.37 (m, 1H), 2.32 (m, 1H), 2.28 – 2.23 (m,2H), 2.19 (t, J = 7.1 Hz, 1H), 1.75 (s, 3H), 1.19 (d, J = 7.6 Hz, 3H). 13C NMR (151MHz, CDCl3) δ 218.48, 169.52, 142.06, 137.56, 133.57, 123.04, 118.88, 113.63,82.01, 77.23, 77.02, 76.81, 49.36, 43.45, 43.08, 42.18, 41.27, 37.71, 22.03,17.35. IR (neat): 3082, 2926, 1760, 1739, 1641 cm -1 HRMS(ESI) m / z: [M+H] + calculated for C 17 H 22 O3275.1641; found 275.1641.

[0084] 2.6 Preparation of Compound III-9 See also Figure 7 Under a nitrogen atmosphere, compound I-2 (82.9 mg, 0.3 mmol) and 5-fluorouracil (42.9 mg, 0.33 mmol) were added to a 25 mL two-necked flask. 8 mL of THF was then added, and Et3N (60.7 mg, 0.6 mmol, 83.6 μL) was added to the two-necked flask with stirring. The reaction was stirred overnight at room temperature. Completion of the reaction was monitored by TLC. The filtrate was filtered, and the filtrate was concentrated under reduced pressure (using a circulating water multi-purpose vacuum pump at 35°C) to obtain a crude product. The crude product was purified by silica gel column chromatography using a PE:EA ratio of 1:1. The collected eluate was dried under vacuum to obtain 112.9 mg of a colorless solid in a 92.6% yield. The product was identified as compound III-9. HRMS (ESI) m / z: [M+H] + calculated for C 20 H 23 FN2O6407.1540; found 407.1547.

[0085] Considering stability and storage, the prepared compound III-9 was salted as follows: Compound III-9 (81.3 mg, 0.2 mmol) and 5 mL of methanol were added to a 25 mL double-necked flask at room temperature. Then, 2 mL of 0.1 M NaOH solution was added to the flask under stirring. Stirring was continued for 30 minutes before the reaction was terminated. The mixture was extracted with DCM (3 × 3 mL), and the separated aqueous phase was lyophilized to obtain 67.06 mg of a white powdery solid with a yield of 82.5%. The solid was identified as compound III-9 sodium salt (abbreviated as III-9-Na). HRMS (ESI) m / z: [M-Na] - calculated for C 20 H 22 FN2NaO6405.1462;found405.1466.

[0086] 2.7 Preparation of Compound II-1 See also Figure 8 To a 25 mL double-necked flask at 0 ° C and under a nitrogen atmosphere, compound Ⅰ-1 (55.3 mg, 0.2 mmol) and 5 mL of THF were added. NaH (60% dispersion, 8 mg, 0.2 mmol; alkaline reagent for reacting with hydroxyl to form sodium alcohol) was added to the double-necked flask under stirring, followed by reflux reaction at 85 ° C for 30 min. Then, propargyl bromide (28.6 mg, 0.24 mmol, 20.7 μL) was added to the double-necked flask, and the reaction was continued at 85 ° C for 10 h. After the reaction was completed, it was cooled to room temperature, 10 drops of water were added to the reaction system, and stirring was continued for 30 min to quench the reaction. The solvent was then removed under reduced pressure (25-35 ° C, 0.08-0.09 MPa) to obtain a yellow oily crude product. The crude product was purified by silica gel column chromatography using a mixture of petroleum ether (60-90°C) and ethyl acetate (3:1). The collected eluent was dried under vacuum to yield 56.3 mg of a white solid with a yield of 89.6%. This solid was identified as Compound II-1′. HRMS (ESI) m / z: [M+H] + calculated for C 20 H 26 O3315.1882; found 315.1887.

[0087] Dimethylamine hydrochloride (24.5 mg, 0.3 mmol) and 10 mL of methanol were placed in a 50 mL two-necked flask and stirred at room temperature for 30 min. Then, TEA (0.4 mmol, 56 μL) was added dropwise to the two-necked flask using a syringe. After the addition was complete, the mixture was vigorously stirred for 30 min under nitrogen protection. Then, compound II-1' (62.9 mg, 0.2 mmol) was added to the two-necked flask, and then stirred at room temperature overnight. The reaction was monitored by TLC. The methanol was removed under reduced pressure, and 10 mL of dichloromethane was added to the residue to dissolve it. The residue was washed with 5 mL of citric acid solution and saturated brine in sequence, and then dried over anhydrous Na2SO4. The residue was concentrated under reduced pressure (25-35 °C, 0.08-0.09 MPa) to obtain 52.1 mg of a white solid (i.e., compound II-1) with a yield of 72.4%.

[0088] Considering the stability and storage convenience, the prepared compound II-1 is salified according to the following steps: Figure 9 At room temperature, compound II-1 (71.9 mg, 0.2 mmol) and 5 mL of dichloromethane were added to a 25 mL round-bottom flask and stirred until compound II-1 was completely dissolved. 0.1 M HCl solution was then slowly added dropwise to the round-bottom flask until the pH of the solution reached approximately 3. 5 mL of purified water was then added to the round-bottom flask and stirred for 5 minutes. The organic phase was separated using a separatory funnel and the aqueous phase was extracted with DCM (3 × 5 mL). The extracted aqueous phase was freeze-dried to yield 66.1 mg of a white powdery solid with a yield of 83.4%. The product was identified as compound II-1 hydrochloride (abbreviated as II-1-HCl). HRMS (ESI) m / z: [M-Cl] + calculated for C 22 H 34 ClNO3360.2533; found 360.2529.

[0089] 2.8 Preparation of Compound IV-2 See also Figure 10To a 25 mL two-necked flask at 0°C under a nitrogen atmosphere, compound 8a (38.1 mg, 0.1 mmol), AcO (12.3 mg, 0.12 mmol, 11.24 μL), DMAP (9.8 mg, 0.08 mmol), and 1 mL of pyridine were added. The mixture was stirred for 30 min, and the reaction system was then allowed to stand at room temperature and stirred for 12 h under a nitrogen atmosphere. After confirming complete reaction of the starting materials by TLC, 10 drops of water were added to the reaction system, and stirring was continued for 30 min. The reaction was then quenched by adding 20 mL of ethyl acetate, dried over anhydrous NaSO, filtered, and the filtrate was concentrated under reduced pressure (using a circulating water multi-purpose vacuum pump at 35°C) to obtain the crude product. The crude product was purified by silica gel column chromatography using a ratio of petroleum ether (60-90°C):ethyl acetate (10:1). The collected eluate was evaporated to vacuum to afford 39.2 mg of a light yellow oil in a yield of 95.6%. The product was identified as compound Ⅳ-2', HRMS (ESI) m / z: [M+H] + calculated for C 23 H 38 O5Si 423.2489;found423.2495.

[0090] Under nitrogen atmosphere and 0 ° C, compound IV-2' (43.2 mg, 0.1 mmol) and 5.0 mL of anhydrous THF were added to a 20 mL reaction flask containing a magnetic stirrer. Then, 0.25 mL of TBAF (0.25 mmol, 2.5 equiv.) was added (slowly dropwise) to the reaction flask. The mixture was then transferred to room temperature and stirred under nitrogen atmosphere for 2 h. After TLC monitoring with eluent (PE:EA=3:1) confirmed that the raw material reaction was complete, 10 mL of water was added to terminate the reaction. The reaction was then extracted with ethyl acetate (3×10 mL). The organic phases were combined and washed with 20 mL of saturated brine, then dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure (25-35 ° C, 0.08-0.09 MPa) to obtain the crude product. The crude product was purified by silica gel column chromatography using PE:EA=3:1 as the eluent. The collected eluent was dried under vacuum to obtain 25.1 mg of a white solid with a yield of 81.3%. The solid was identified as compound IV-2. HRMS (ESI) m / z: [M+H] + calculated for C 17 H 24 O5309.1624; found 309.1629.

[0091] 2.9 Preparation of Compound V-22 See also Figure 11 To a 25-mL double-necked flask at 0°C under a nitrogen atmosphere, compound 8b (79.1 mg, 0.2 mmol) and 5 mL of THF were added. NaH (60% dispersion, 8 mg, 0.2 mmol) was added to the double-necked flask with stirring, and the mixture was refluxed at 85°C for 30 min. Propargyl bromide (28.6 mg, 0.24 mmol, 20.7 μL) was then added to the double-necked flask, and the mixture was refluxed at 85°C for 10 h. After the reaction was completed, the mixture was cooled to room temperature, 10 drops of water were added to the reaction system, and stirring was continued for 30 min to quench the reaction. The solvent was then removed under reduced pressure (25-35°C, 0.08-0.09 MPa) to obtain a crude yellow oily product. The crude product was purified by silica gel column chromatography using PE (60-90°C):EA = 10:1 as the eluent. The collected eluent was dried under vacuum to yield 74.8 mg of a colorless oil with a yield of 86.4%. This product was identified as Compound V-22′. HRMS (ESI) m / z: [M+H] + calculated for C 25 H 40 O4Si433.2696; found 433.2691.

[0092] Under nitrogen atmosphere and 0 ° C, compound V-22' (43.4 mg, 0.1 mmol) and 5.0 mL of anhydrous THF were added to a 20 mL reaction bottle containing a magnetic stirrer. Then, 0.25 mL of TBAF (0.25 mmol, 2.5 equiv.) was added (slowly dropwise) to the reaction bottle. The mixture was then transferred to room temperature and stirred under nitrogen atmosphere for 2 h. After TLC monitoring with eluent (PE:EA=3:1) confirmed that the reaction of the raw materials was complete, 10 mL of water was added to terminate the reaction, and then extracted with ethyl acetate (3×10 mL). The organic phases were combined and washed with 20 mL of saturated brine, then dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure (25-35 ° C, 0.08-0.09 MPa) to obtain the crude product. The crude product was purified by silica gel column chromatography using a PE:EA ratio of 3:1. The collected eluate was dried under vacuum to obtain 27.9 mg of a white solid compound with a yield of 87.4%. The compound was identified as compound V-22. HRMS (ESI) m / z: [M+H] + calculated for C 19 H 26 O4319.1831; found 319.1836.

[0093] 2.10 Preparation of Compound VI-4 See also Figure 12 Under a nitrogen atmosphere, compound I-5 (54.9 mg, 0.2 mmol) and 4-trifluoromethylpiperidine (33.7 mg, 0.22 mmol, 30.6 μL) were added to a 25 mL two-necked flask. 8 mL of THF was then added, and KCO (55.3 mg, 0.4 mmol) was added to the flask with stirring. The mixture was stirred overnight at room temperature. The reaction was monitored by TLC, filtered, and the filtrate was concentrated under reduced pressure (using a circulating water multi-purpose vacuum pump at 35°C) to obtain a crude product. The crude product was purified by silica gel column chromatography using a PE:EA ratio of 2:1. The collected eluate was dried under vacuum to obtain 76.7 mg of a colorless solid with a yield of 89.7%. The product was identified as compound VI-4. HRMS (ESI) m / z: [M+H] + calculated for C 23 H 32 F3NO3428.2334; found 428.2337.

[0094] (III) Preparation of Tablets of 1,10-Split-ring Guaiane Sesquiterpenoid Lactone Pseudo-Natural Products and Their Derivatives 300 mg of compound Ⅰ-1, Ⅰ-2, Ⅰ-3, Ⅰ-4, Ⅰ-5, IV-2, Ⅴ-22, Ⅵ-4, Ⅲ-9-Na or Ⅱ-1-HCl were mixed with 2.625 g of excipients (white dextrin: lactose = 7:3, mass ratio), 95% ethanol was added, granulated, dried, sieved, 0.075 g of sodium stearate was added, and the mixture was mixed evenly and tableted (each tablet weighing 200 mg) to obtain tablets containing 20 mg of compound Ⅰ-1, Ⅰ-2, Ⅰ-3, Ⅰ-4, Ⅰ-5, IV-2, Ⅴ-22, Ⅵ-4, Ⅲ-9-Na or Ⅱ-1-HCl.

[0095] (IV) Preparation of 1,10-Split-ring Guaiane Sesquiterpenoid Lactone Pseudo-Natural Products and Their Derivatives for Injection Dissolve 150 mg of compound Ⅰ-1, Ⅰ-2, Ⅰ-3, Ⅰ-4, Ⅰ-5, IV-2, Ⅴ-22, Ⅵ-4, Ⅲ-9-Na or Ⅱ-1-HCl and 750 mg of mannitol in 17 mL of water for injection. After initial mixing, adjust the volume to 30 mL. Filter the resulting solution and fill it into vials (1 mL per bottle). Lyophilize, seal and sterilize to obtain a lyophilized powder injection containing 5 mg of compound Ⅰ-1, Ⅰ-2, Ⅰ-3, Ⅰ-4, Ⅰ-5, IV-2, Ⅴ-22, Ⅵ-4, Ⅲ-9-Na or Ⅱ-1-HCl per vial.

[0096] (V) Preparation of capsules of 1,10-split-ring guaiacyl sesquiterpenoid lactone pseudo-natural products and their derivatives 150 mg of compound Ⅰ-1, Ⅰ-2, Ⅰ-3, Ⅰ-4, Ⅰ-5, IV-2, Ⅴ-22, Ⅵ-4, Ⅲ-9-Na or Ⅱ-1-HCl was mixed with 1.35 g of excipients (white dextrin: lactose = 7:3, mass ratio), 95% ethanol was added, granulated, dried, sieved, and loaded into capsules (each capsule weighing 150 mg) to obtain capsules containing 15 mg of compound Ⅰ-1, Ⅰ-2, Ⅰ-3, Ⅰ-4, Ⅰ-5, IV-2, Ⅴ-22, Ⅵ-4, Ⅲ-9-Na or Ⅱ-1-HCl.

[0097] (VI) Experimental study on the inhibition of erythrocyte hemolysis by compounds Ⅰ-1, Ⅰ-2, Ⅰ-3, Ⅰ-4, Ⅰ-5, Ⅱ-1, Ⅲ-9, Ⅳ-2, Ⅴ-22 and Ⅵ-4 6.1 Experimental Materials (1) Experimental reagents: Plasma free hemoglobin assay kit (Railda).

[0098] (2) Experimental cells: human red blood cells (from the Xi'an Blood Center of the Chinese People's Liberation Army); fresh blood collected on the same day was placed in a centrifuge tube with an anticoagulant and centrifuged to obtain two layers, the upper layer being plasma and the lower layer being red blood cells. The red blood cells in the lower layer were removed for the experiment.

[0099] (3) Experimental instruments: enzyme-labeled analyzer, etc.

[0100] 6.2 Experimental plan For the red blood cell hemolysis model, hemolysis refers to the rupture or destruction of red blood cells (RBCs), resulting in the release of hemoglobin and other cellular contents into the surrounding environment.

[0101] Erythrocytes were incubated with various concentrations of small molecule compounds (0 µM, 2 µM, 4 µM, 8 µM, and 16 µM) at 37°C for 30 min. Compounds I-1, I-2, I-3, I-4, I-5, II-1, III-9, IV-2, V-22, VI-4, or MCC950 (as a positive control) were then stimulated with nigericin at a final concentration of 1 µM for 50 min (i.e., the N+0 µM, N+2 µM, N+4 µM, N+8 µM, and N+16 µM groups in Table 1). In the model group (i.e., the N+0 µM group in Table 1), nigericin induced pyroptosis, leading to cell death and hemoglobin efflux. After treatment, hemoglobin was quantified using a kit. The activity of the small molecule compound was determined based on the proportionality between hemoglobin content and absorbance (OD505).

[0102] Table 1. Experimental groups and treatments

[0103] Note: In Table 1, the total volume of each group is 200 μL, N represents Nigericin, the final concentration of Nigericin in the system is 1 μM, X represents compounds Ⅰ-1, Ⅰ-2, Ⅰ-3, Ⅰ-4, Ⅰ-5, Ⅱ-1, Ⅲ-9, Ⅳ-2, Ⅴ-22, Ⅵ-4 or MCC950 (the solvent for preparing the solution is 0.9% saline), and NS represents 0.9% saline.

[0104] 6.3 Experimental steps S1: Pipette 1 mL of normal saline (0.9% NaCl) and add it to a 1.5 mL EP tube.

[0105] S2: Pipette 100 µL of the lower layer of red blood cells (i.e., the red blood cells separated from the blood) and add them to an EP tube containing 1.5 mL of normal saline.

[0106] S3 was centrifuged at 3000 rpm for 5 min.

[0107] S4 After centrifugation, discard 1 mL of supernatant, add 1 mL of normal saline, and centrifuge again at 3000 r / min for 5 min.

[0108] S5 After centrifugation, the supernatant is discarded and the remaining is the required red blood cells (the red blood cell washing is now completed).

[0109] S6 Prepare suspensions of physiological saline and red blood cells according to the 7 groups of schemes in Table 1.

[0110] S7 Add the small molecule compound to be tested into the red blood cell suspension of the corresponding group at 0 µL, 32 µL, 0 µL, 4 µL, 8 µL, 16 µL, and 32 µL, respectively (so that the final concentrations of the small molecule compound in the Control, 16 µM, N+0 µM, N+2 µM, N+4 µM, N+8 µM, and N+16 µM groups are: 0 µM, 16 µM, 0 µM, 2 µM, 4 µM, 8 µM, and 16 µM, respectively) and incubate at 37°C for 30 min.

[0111] After the S8 incubation, 4 μL of Nigericin was added to the N+0 μM, N+2 μM, N+4 μM, N+8 μM, and N+16 μM groups and incubated at 37 °C for 50 min.

[0112] After the S9 incubation, the cells were centrifuged at 2000 rpm for 3 min.

[0113] S10: Aspirate the supernatant and set aside.

[0114] S11 Detect hemoglobin in the supernatant using a plasma free hemoglobin assay kit (detection conditions: react at 37°C for 20 min after adding the reagent).

[0115] After the S12 reaction was completed, the absorbance at a wavelength of 505 nm was measured using a microplate reader.

[0116] 6.4 Experimental Results like Figure 13 As shown in the experimental results, using the release of hemoglobin from erythrocytes stimulated with nigericin after 30 minutes of co-incubation with MCC950 as a reference (absorbance detection), compounds I-1 (the absorbance value decreased with increasing I-1 concentration, indicating that the concentration of free hemoglobin gradually decreased, indicating that erythrocyte hemolysis was alleviated with increasing I-1 concentration), I-3, II-1 (a comparison of the absorbance values ​​of II-1 at different concentrations revealed that its absorbance value decreased with increasing drug concentration, indicating that II-1 could reduce hemoglobin concentration and thus inhibit erythrocyte hemolysis), III-9, IV-2, and VI-4 all exhibited significant inhibitory effects on erythrocyte hemolysis induced by the induced activation reagent nigericin at a treatment concentration of 4 µM, with inhibitory effects superior to or comparable to those of MCC950. Compounds I-1 and II-1 exhibited the greatest inhibitory effects at treatment concentrations of 2 µM and 4 µM, respectively. These results suggest that 1,10-split-ring guaiacyl sesquiterpene lactone compounds (or 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural products) represented by compound Ⅰ-3 and compounds derived therefrom have a certain inhibitory effect on erythrocyte hemolysis and can be used as candidate drug molecules for preventing and / or treating NLRP3 inflammasome-related diseases.

[0117] Figure 13 The results of each parallel experiment in the N+0 μM group fluctuated (OD505 values ​​were high and low), which may be due to the differences in the red blood cells used in the experiments with different small molecule compounds.

[0118] In summary, based on the synthesis of 1,10-split-ring guaiacyl sesquiterpenoid lactone pseudo-natural products and their derivatives, the present invention discovered novel 1,10-split-ring guaiacyl sesquiterpenoid lactone compounds that have a significant inhibitory effect on erythrocyte hemolysis. These compounds have high application value in the development of anti-inflammatory drugs (especially anti-inflammatory drugs for the prevention and / or treatment of NLRP3-related inflammatory diseases such as inflammatory bowel disease) (e.g., large-scale synthesis and screening of drug molecules with anti-inflammatory activity, and use as active ingredients in the preparation of anti-inflammatory drugs).

Claims

1. A 1,10-split-ring guaiacyl sesquiterpene lactone compound, characterized in that: The compound is a compound having a structure as shown in Formula I or any one of its pharmaceutically acceptable salts, tautomers, stereoisomers, and precursor compounds: In Formula I: R1 and R2 together form a double bond; or R1 is hydrogen and R2 is selected from any one of an alkylaminomethyl group, an aromatic aminomethyl group, a heteroaromatic aminomethyl group, and a group having a structure as shown in Formula X: R3, R4 and the N atom in formula X form a cyclic structure, and the cyclic structure is a heterocyclic ring containing one N atom; R5 is allyl, acetaldehyde, -C2H4ORc, -C2H4OCOORc, -C2H4OCORc, -C2H4OCONRcRd, -CH2ORc, -CH2OCOORc, -CH2OCORc or -CH2OCONRcRd; R6 is hydrogen; R7 is a methyl group; R8 is a carbonyl oxygen atom, -ORc, -OCOORc, -OCORc or -OCONRcRd; The Rc and Rd are each selected from any one of hydrogen, alkynyl, cycloalkyl, halogen, alkyl, aryl, halogen-substituted alkyl, halogen-substituted aryl, and nitro-substituted aryl.

2. A 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product, characterized in that: The pseudo-natural product is a compound having a structure as shown in Formula I-1, Formula I-2, Formula I-3, Formula I-4 or Formula I-5, or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers, and precursor compounds of the compound: 。 3. A 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product derivative, characterized by: The pseudo-natural product derivative is a compound having a structure as shown in Formula II, Formula III, Formula IV, Formula V or Formula VI, or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers, and precursor compounds of the compound: Among them, R9, R 10 are selected from any one of Rc, COORc, CORc, and CONRcRd, wherein Rc and Rd are selected from any one of hydrogen, alkynyl, cycloalkyl, halogen, alkyl, aryl, halogen-substituted alkyl, halogen-substituted aryl, and nitro-substituted aryl; R 11 Any one selected from alkylamino, arylamino, heteroarylamino, and heterocyclic amino containing one nitrogen atom.

4. A method for preparing a 1,10-split-ring guaiacyl sesquiterpene lactone compound, characterized in that: The following steps are involved: by( R )-Carvone as a raw material, a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product with a protected hydroxyl group structure as shown in Formula I-3 or Formula I-4 is prepared by chemical synthesis, and a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product with a protected hydroxyl group structure as shown in Formula I-4 is prepared by using the 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product with a protected hydroxyl group structure as shown in Formula I-1, and then a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product with a protected hydroxyl group structure as shown in Formula I-1, Formula I-3 or Formula I-4 is prepared by deprotection reaction respectively: ； Then, the 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural products with structures such as those shown in Formula I-1 and Formula I-4 are used to prepare the 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural products with structures such as those shown in Formula I-5 and Formula I-2 respectively through oxidation reactions: 。 5. A method for preparing a 1,10-split-ring guaiacyl sesquiterpene lactone compound, characterized in that: The following steps are involved: 1) allowing a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product having a structure as shown in Formula I-1 to undergo a condensation reaction with a carboxylic acid, anhydride, or an acyl halide in the presence of a condensing agent, a catalyst, and a base, or in the presence of a catalyst and a base, to produce an intermediate containing an acyl group or an ether bond; Alternatively, a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product having a structure as shown in Formula I-1 is subjected to a nucleophilic substitution reaction with a halogenated hydrocarbon to prepare an intermediate containing an ether bond; The 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product with the structure shown in Formula I-1 is: ； 2) Under alkaline conditions, the intermediate prepared in step 1 is subjected to a selective intermolecular Michael addition reaction with an organic amine to obtain a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product derivative having a structure as shown in Formula II: wherein R9 is selected from any one of Rc, COORc, CORc, and CONRcRd, and Rc and Rd are independently selected from any one of hydrogen, alkynyl, cycloalkyl, halogen, alkyl, aryl, halogen-substituted alkyl, halogen-substituted aryl, and nitro-substituted aryl; R 11 Any one selected from alkylamino, arylamino, heteroarylamino, and heterocyclic amino containing one nitrogen atom.

6. A method for preparing a 1,10-split-ring guaiacyl sesquiterpene lactone compound, characterized in that: The following steps are involved: Under alkaline conditions, a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product having a structure as shown in Formula I-2 or Formula I-5 is used as a raw material: ； The raw materials are subjected to a selective intermolecular Michael addition reaction with an organic amine to prepare a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product derivative having a structure as shown in Formula III or Formula VI: Among them, R 11 Any one selected from alkylamino, arylamino, heteroarylamino, and heterocyclic amino containing one nitrogen atom.

7. A method for preparing a 1,10-split-ring guaiacyl sesquiterpene lactone compound, characterized in that: The following steps are involved: 1) allowing a hydroxyl-protected 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product to undergo a condensation reaction with a carboxylic acid, anhydride, or an acyl halide in the presence of a condensing agent, a catalyst, and a base, or in the presence of a catalyst and a base, to produce an intermediate containing an acyl group or an ether bond; Alternatively, a hydroxyl-protected 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product is subjected to a nucleophilic substitution reaction with a halogenated hydrocarbon to prepare an intermediate containing an ether bond; The hydroxyl-protected 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product is any one of the following structures: ； 2) subjecting the intermediate prepared in step 1 to a deprotection reaction to obtain a 1,10-split-ring guaiacyl sesquiterpene lactone pseudo-natural product derivative having a structure as shown in Formula IV or Formula V: Among them, R 10 Any one selected from Rc, COORc, CORc, CONRcRd, wherein Rc and Rd are independently selected from any one of hydrogen, alkynyl, cycloalkyl, halogen, alkyl, aryl, halogen-substituted alkyl, halogen-substituted aryl, and nitro-substituted aryl.

8. An anti-inflammatory drug, characterized in that: The medicine comprises the 1,10-split ring guaiacyl sesquiterpene lactone compound as claimed in claim 1.

9. A drug for preventing and / or treating NLRP3 inflammasome-related diseases, characterized in that: The medicine comprises the 1,10-split ring guaiacyl sesquiterpene lactone compound as claimed in claim 1.

10. Use of the 1,10-split-ring guaiacyl sesquiterpene lactone compound according to claim 1 in the preparation of anti-inflammatory drugs and drugs for preventing and / or treating NLRP3 inflammasome-related diseases.

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