Guaiane-type sesquiterpene lactone pseudo-natural product, and preparation method therefor and use thereof

Abstract

Disclosed in the present invention are a guaiane-type sesquiterpene lactone pseudo-natural product, and a preparation method therefor and the use thereof. The pseudo-natural product and a derivative thereof are prepared by means of a chemical synthesis method using (R)-Carvone as a starting material. Experiments show that the guaiane-type sesquiterpene lactone compound prepared in the present invention exhibits an inhibitory effect on erythrocyte hemolysis, can be used as a candidate drug molecule for preventing and / or treating diseases associated with the NLRP3 inflammasome, and also has the potential for development as a related anti-inflammatory drug.

Description

A guaiacane-type sesquiterpene lactone pseudonatural product, its preparation method and uses Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to the preparation of guaiacol-type sesquiterpene lactone pseudo-natural products and their derivatives by chemical synthesis using (R)-Carvone (i.e., L-carvone) as the starting material, and the testing of the anti-inflammatory activity of these compounds on an established erythrocyte hemolysis model. Background Technology

[0002] The NLRP3 inflammasome, an organelle structure first discovered and reported in 2002 by the Biomedical Research Center at the University of Lausanne, Switzerland, consists of three parts: the NLRP3 scaffold, the adaptor protein ASC, and Caspase-1. Studies have shown that when the body responds to pathogen infection or danger-related signals, the NLRP3 inflammasome assembles and activates in the cytoplasm, leading to the activation and secretion of pro-inflammatory factors IL-1β and IL-18, thus playing an important role in innate immune-mediated inflammation. Furthermore, the NLRP3 inflammasome is closely related to the development of many major human diseases, including type 2 diabetes (T2D), cyropyrin-associated periodic syndrome (CAPS), gout, multiple sclerosis, Alzheimer's disease (AD), Parkinson's disease (PD), and inflammatory bowel disease (IBD). Therefore, NLRP3 is also an important target for the development of drugs to treat inflammation-related diseases; however, no drugs have been reported since its discovery. Taking IBD as an example, the main clinical drugs used to treat IBD are steroidal anti-inflammatory drugs such as glucocorticoids, nonsteroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen, and immunosuppressants such as azathioprine. In addition, biological agents such as TNF-α monoclonal antibodies can also be used for treatment.

[0003] Sesquiterpene lactones exhibit significant anti-inflammatory potential by targeting and inhibiting various key pro-inflammatory molecules involved in different inflammatory pathways and preventing or reducing inflammatory damage to tissues. Therefore, sesquiterpene lactones are an important source of novel anti-inflammatory drugs. However, for known guaiacol-type sesquiterpene lactones, anti-inflammatory activity has only been verified in a few synthesized derivatives (see: CN117624098A, CN113968869A, etc.), and no novel parent or core structures of guaiacol-type sesquiterpene lactones that can meet the needs of large-scale drug molecule synthesis and screening have yet been discovered.

[0004] Peudo-natural products (PNPs) are a concept first proposed in 2018 by the Max Planck Institute for Molecular Physiology in Germany as part of a natural product chemistry optimization strategy. Currently, there are no research reports on the use of guaiacane-type sesquiterpene lactone pseudo-natural products and their derivatives as anti-inflammatory drugs, especially as anti-inflammatory drugs specifically targeting the NLRP3 inflammasome. Summary of the Invention

[0005] The purpose of this invention is to provide a guaiacol-type sesquiterpene lactone pseudonatural product, its preparation method, and its uses.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] In a first aspect, a guaiacane-type sesquiterpene lactone compound is provided, wherein the compound is any one of the following: a compound with the structure shown in Formula I, or a pharmaceutically acceptable salt, tautomer, stereoisomer, or precursor compound of the compound:

[0008] In Formula I:

[0009] The double bond is formed by R1 and R2, and R9 is hydrogen; or the double bond is formed by R9 and R2, and R1 is methyl; or both R9 and R1 are hydrogen and R2 is selected from any one of the following groups: alkylaminomethyl, arylaminomethyl, heteroarylaminomethyl, or the group with the structure shown in the following formula (i.e., formula X):

[0010] In formula X, R3, R4, and N atoms form a cyclic structure (specifically, a heterocyclic ring containing one N atom and the rest being C atoms), which is a 3- to 7-membered ring. The hydrogen atoms in the cyclic structure (which can refer to the aforementioned 3- to 7-membered rings, or the cyclic structures in the aforementioned arylaminomethyl and heteroarylaminomethyl groups) are either unsubstituted or substituted by one or more groups (if substitution occurs, the corresponding substituent group can be selected from one or more of alkyl, cycloalkyl, alkenyl, alkynyl, arylaryl, arylalkyl, arylalenyl, arylalkynyl, heterocyclic, carboxyl, nitro, halogen, carbonyl oxygen atom, sulfur atom, and halogen-substituted alkyl groups).

[0011] R5 is hydrogen;

[0012] R6 is a methyl group;

[0013] R7 is a carbonyl oxygen atom, -ORc, -OCOORc, -OCORc, or -OCONRcRd;

[0014] R8 can be hydrogen, -ORc, -OCOORc, -OCORc, or -OCONRcRd;

[0015] 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.

[0016] Secondly, a guaiacane-type sesquiterpene lactone pseudonatural product is provided, wherein the pseudonatural product is any one of the following: a compound with the structure shown in Formula I-1, Formula I-2 or Formula I-3, or a pharmaceutically acceptable salt, tautomer, stereoisomer or precursor compound of the compound:

[0017] Among them, the guaiacane-type sesquiterpene lactone pseudo-natural product shown in Formula I-1 is the compound corresponding to the compound shown in Formula I with R1 and R2 forming a double bond, R9 being hydrogen, R7 being a carbonyl oxygen atom, and R8 being hydrogen; the guaiacane-type sesquiterpene lactone pseudo-natural product shown in Formula I-2 is the compound corresponding to the compound shown in Formula I with R9 and R2 forming a double bond, R1 being methyl, R7 being hydroxyl, and R8 being hydroxyl; the guaiacane-type sesquiterpene lactone pseudo-natural product shown in Formula I-3 is the compound corresponding to the compound shown in Formula I with R1 and R2 forming a double bond, R9 being hydrogen, R7 being hydroxyl, and R8 being hydrogen.

[0018] Thirdly, a guaiacol-type sesquiterpene lactone pseudo-natural product derivative is provided, wherein the pseudo-natural product derivative is a compound with the structure shown in Formula II, Formula III or Formula V (the structure of these compounds also conforms to the general formula I above) or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers or precursor compounds of the compound:

[0019] Among them, R 10 Rc is selected from any one of Rc, COORc, CORc, and CONRcRd, wherein Rc and Rd are each independently selected from any one of alkynyl, cycloalkyl, halogen, alkyl, aryl, halogen-substituted alkyl, halogen-substituted aryl, and nitro-substituted aryl; R 11 The cyclic structure is selected from any one of alkylamino, aromaticamino, heteroaromaticamino, and heterocyclic amino containing one nitrogen atom, wherein the heterocyclic containing one nitrogen atom is a 3- to 7-membered ring; the hydrogen atoms in the cyclic structure (which can refer to the aforementioned 3- to 7-membered rings, or the cyclic structures in the aforementioned aromaticamino and heteroaromaticamino) are either unsubstituted or substituted by one or more groups (if substitution occurs, the corresponding substituent group can be selected from one or more of alkyl, cycloalkyl, alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, arylalenyl, arylalkynyl, heterocyclic, carboxyl, nitro, halogen, carbonyl oxygen atom, sulfur atom, and halogen-substituted alkyl).

[0020] Preferably, the R 11It is selected from any one of dimethylamino, pyrrolyl, 2-carboxy-pyrrolyl, piperidinyl, 4-trifluoromethyl-piperidinyl, 5-nitro-indazole, imidazolyl, 4-nitro-imidazolyl, 1,2,4-triazolyl, 1,2,3-triazolyl, 5-fluorouracilyl, 6-mercaptopurine, 6-nitroindazole, and pyrazolyl.

[0021] Preferably, Rc or Rd is selected from any one of the following: C1-C10 (i.e., 1-10 carbon atoms) alkyl groups (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-octyl, n-nonyl, etc.), C3-C7 (i.e., 3-7 carbon atoms) cycloalkyl groups (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc.), ethynyl, 2-propynyl, 3-butynyl, phenyl, C1-C6 (i.e., 1-6 carbon atoms) halogen-substituted alkyl groups (e.g., chloromethyl, bromomethyl, chloroethyl, bromoethyl, fluoroethyl, chlorobutyl, etc.), halogen-substituted phenyl groups (e.g., 4-fluoro-phenyl, 4-chloro-phenyl, 4-bromo-phenyl, etc.), nitro-substituted phenyl groups (e.g., 4-nitro-phenyl, etc.), and halogens (e.g., chlorine, etc.).

[0022] Preferably, the pseudo-natural product derivative is a compound with the structure shown in Formulas II-1 to II-12, or any one of the pharmaceutically acceptable salts, tautomers, stereoisomers, or precursor compounds of such compound:

[0023] Preferably, the pseudo-natural product derivative is a compound with the structure shown in Formulas III-1 to III-12, or any one of a pharmaceutically acceptable salt, tautomer, stereoisomer, or precursor compound of the compound:

[0024] Preferably, the pseudo-natural product derivative is any one of the following: a compound with the structure shown in formulas V-1 to V-31, or a pharmaceutically acceptable salt, tautomer, stereoisomer, or precursor compound of the compound:

[0025] Fourthly, a method for preparing guaiacane-type sesquiterpene lactone compounds is provided, comprising the following steps:

[0026] Using (R)-Carvone (i.e., L-carvone) as a starting material, guaiacane sesquiterpene lactones with hydroxyl protected structures, as shown in Formula I-3 above, were prepared through chemical synthesis (including reacting a cyclopentane-1-carboxaldehyde intermediate with protected hydroxyl groups synthesized from L-carvone with an intermediate containing alcoholic hydroxyl groups and a methylene chain synthesized from butynol, to synthesize 1,10-secrosic guaiacane sesquiterpene lactones, followed by oxidation of the corresponding alcoholic hydroxyl groups in the structure of the compound to aldehyde groups and conversion of the generated aldehyde groups to alkenyl groups, and then catalytic cyclization using a Grubb II catalyst). The ester-type pseudo-natural product (hereinafter referred to as hydroxyl-protected compound I-3) is used to prepare a guaiacane-type sesquiterpene lactone pseudo-natural product (hereinafter referred to as compound I-3) with the structure shown in Formula I-3 above. Then, using the guaiacane-type sesquiterpene lactone pseudo-natural product with the structure shown in Formula I-3, a guaiacane-type sesquiterpene lactone pseudo-natural product (hereinafter referred to as compound I-1) with the structure shown in Formula I-1 above is prepared by an oxidation reaction. The specific structure of hydroxyl-protected compound I-3 is shown in the following formula (in the following formula, "PG" refers to a protecting group formed by any of the reagents that can be used to protect the corresponding hydroxyl group, such as TBS, etc.):

[0027] Alternatively, using (R)-Carvone (i.e., L-carvone) as a raw material, a guaiac-type sesquiterpene lactone compound with a protected hydroxyl group synthesized from L-carvone and an intermediate with a retained alcohol hydroxyl group and methylene chain synthesized from propynyl alcohol are reacted to synthesize a 1,10-secrosic guaiac-type sesquiterpene lactone compound as shown in Formula I-2 above (hereinafter referred to as hydroxyl-protected compound I-2) is prepared by chemical synthesis (including the reaction of a cyclopentane-1-carboxaldehyde intermediate with a protected hydroxyl group synthesized from L-carvone and an intermediate with a retained alcohol hydroxyl group and methylene chain synthesized from propynyl alcohol, followed by catalytic cyclization using a second-generation Grubb catalyst).

[0028] Preferably, in the oxidation reaction, dichloromethane (DCM), tetrahydrofuran (THF), or acetonitrile is used as the reaction solvent.

[0029] Preferably, in the oxidation reaction, the molar ratio of compound I-3 to oxidant (e.g., DMP) is 1:1.1 to 1.5, the reaction time is 1 to 5 hours, and the reaction temperature is 0 to 35°C. After the reaction is completed, compound I-1 is obtained by sequential extraction (the solvent added during extraction is dichloromethane or ethyl acetate), washing, drying, filtration, vacuum concentration, and column chromatography.

[0030] Preferably, the mobile phase used in the column chromatography to purify compound I-1 is a petroleum ether (PE) / ethyl acetate (EA) mixed solvent (PE:EA volume ratio = 2 to 5:1).

[0031] Fifthly, a method for preparing guaiacane-type sesquiterpene lactone compounds is provided, comprising the following steps:

[0032] 1) Using (R)-Carvone (i.e., L-carvone) as raw material, a guaiacane-type sesquiterpene lactone intermediate is first prepared through a multi-step chemical reaction. The guaiacane-type sesquiterpene lactone intermediate is a guaiacane-type sesquiterpene lactone pseudo-natural product with the structure shown in Formula I-1 above (hereinafter referred to as Compound I-1), or a guaiacane-type sesquiterpene lactone pseudo-natural product with one hydroxyl group protected as shown in Formula I-2 above (i.e., the above-mentioned hydroxyl-protected Compound I-2), or a guaiacane-type sesquiterpene lactone pseudo-natural product with the structure shown in Formula I-3 above (hereinafter referred to as Compound I-3).

[0033] 2) Chemically synthesize the guaiacol-type sesquiterpene lactone intermediate obtained in step 1 (e.g., through reactions to achieve esterification, etherification and / or intermolecular Michael addition reactions) and prepare guaiacol-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula II, Formula III or Formula V above.

[0034] Preferably, step 2 specifically includes the following steps: under alkaline conditions, reacting compound I-1 with the compound used to provide R. 11 Organic amines undergo selective intermolecular Michael addition reactions to prepare guaiacol-type sesquiterpene lactone pseudo-natural product derivatives (referred to as compound II) with structures as shown in Formula II above.

[0035] Preferably, in the preparation of guaiacane-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula II above, the base used (providing alkaline conditions) is piperidine, triethylamine, sodium hydroxide, potassium hydroxide, sodium ethoxide, potassium carbonate (K2CO3), potassium tert-butoxide, or 1,8-diazabicyclo[5.4.0]undec-7-ene.

[0036] Preferably, in the preparation of guaiacol-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula II above, tetrahydrofuran, acetonitrile, benzene, dichloromethane (DCM) or chloroform are used as reaction solvents.

[0037] Preferably, in the preparation of guaiacane-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula II above, the reaction temperature is 25–35°C, the reaction time is 12–20 h, and compound I-1 is reacted with the compound used to provide the R. 11The molar ratio of the organic amines was 1:1.1 to 2; after the reaction was completed, the mixture was filtered, and the filtrate was concentrated under reduced pressure and subjected to column chromatography to obtain compound II.

[0038] Preferably, the mobile phase used in the column chromatography for purifying compound II is a petroleum ether (PE) / ethyl acetate (EA) mixed solvent (PE:EA volume ratio = 1 to 3:1).

[0039] Preferably, step 2 specifically includes the following steps: reacting the hydroxyl-protected compound I-2 with a condensing agent, a catalyst, and a base (or a catalyst and a base) in the presence of a condensing agent, a catalyst, and a base to provide the R. 10 The carboxylic acid, anhydride, or acyl halide (e.g., acyl chloride) undergoes a condensation reaction (in the condensation reaction, if the anhydride is highly reactive, the condensing agent can be omitted), followed by a deprotection reaction, thereby preparing a guaiacane-type sesquiterpene lactone pseudo-natural product derivative (referred to as compound III) with the structure shown in Formula III above; wherein, the structure of hydroxyl-protected compound I-2 is specifically shown in the following formula (in the following formula, "PG" refers to a protecting group formed by any of the reagents that can be used to protect the corresponding hydroxyl group, such as TBS, etc.):

[0040] Alternatively, step 2 specifically includes the following steps: reacting the hydroxyl-protected compound I-2 (structure as shown above) with the compound used to provide the R 10 The haloalkanes undergo nucleophilic substitution reactions, followed by deprotection reactions, to prepare guaiacol-type sesquiterpene lactone pseudo-natural product derivatives (referred to as compound III) with structures as shown in Formula III above.

[0041] Preferably, in preparing guaiacol-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula III above, the condensing agent used is N,N′-dicyclohexylcarbodiimide (DCC), carbonyl diimidazole (CDI), diisopropylcarbodiimide (DIC), or 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), and the molar ratio of hydroxyl-protected compound I-2 to the condensing agent is 1:1.1-2; the catalyst used is 4-dimethylaminopyridine (DMAP) or 1-hydroxybenzotriazole (HOBt), and the molar ratio of hydroxyl-protected compound I-2 to the catalyst is 1:0.5-0.8; the base used is triethylamine, pyridine (Py), sodium hydride, n-butyllithium, or diisopropylaminolithium.

[0042] Preferably, in the preparation of guaiacol-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula III above, the condensation reaction uses tetrahydrofuran, acetonitrile, benzene, dichloromethane, pyridine, or chloroform as the reaction solvent; the nucleophilic substitution reaction uses tetrahydrofuran, diethyl ether, N,N-dimethylformamide, or ethylene glycol dimethyl ether as the reaction solvent.

[0043] Preferably, in the preparation of guaiacane-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula III above, the condensation reaction temperature is 25–35°C, the reaction time is 10–15 h, and the hydroxyl-protected compound I-2 and the compound used to provide the R 10 The molar ratio of the carboxylic acid, acid anhydride, or acyl halide is 1:1.1–2. After the reaction, the mixture is dissolved (using ethyl acetate or dichloromethane as the solvent), dried, filtered, concentrated under reduced pressure, and subjected to column chromatography to obtain hydroxyl-protected compound III. The nucleophilic substitution reaction is carried out at a temperature of 0–35 °C for 2–12 h. The hydroxyl-protected compound I-2 reacts with the compound used to provide the R... 10 The molar ratio of the haloalkanes is 1:1.1 to 2.2. After the reaction is completed (e.g., by quenching the reaction with water), the mixture is successively extracted (the solvent added during extraction is ethyl acetate or dichloromethane), washed, dried, filtered, concentrated under reduced pressure, and column chromatography to obtain compound III with hydroxyl protection.

[0044] Preferably, the mobile phase used in the column chromatography for purifying hydroxyl-protected compound III is a petroleum ether (PE) / ethyl acetate (EA) mixed solvent (PE:EA volume ratio = 5 to 10:1).

[0045] Preferably, the deprotection reaction uses tetrahydrofuran, dichloromethane, acetonitrile, N,N-dimethylformamide (DMF), or 18-crown-6 ether as the reaction solvent, the reaction temperature is 0–35°C, and the reaction time is 2–20 h (e.g., 12–20 h). The molar ratio of the compound to be deprotected (e.g., TBS) (e.g., hydroxyl-protected compound III) to the deprotection reagent (specifically, the reaction reagent) is 1:1.1–2. After the reaction, the compound is sequentially extracted (using ethyl acetate or dichloromethane as the solvent), washed, dried, filtered, concentrated under reduced pressure, and subjected to column chromatography to obtain the target compound (e.g., compound III).

[0046] Preferably, the deprotecting agent is tetrabutylammonium fluoride (TBAF), TBAF / AcOH, 4-methoxyaqueous aldehyde·BF3, TBAF / NH4F, TBAF·(t-BuOH)4, or (Me2N)3S. + F2SiMe3 - (TAS-F), KF, KF·Al2O3, Bu4NCl / KF·H2O, Aq·HF, pyridine·HF (70%) or AcOH / H2O / THF (3:1:1).

[0047] Preferably, the mobile phase used in the column chromatography for purifying the target compound (e.g., compound III) is a petroleum ether (PE) / ethyl acetate (EA) mixed solvent (PE:EA volume ratio = 2 to 5:1).

[0048] Preferably, step 2 specifically includes the following steps: reacting compound I-3 with the compound used to provide the R 10 The condensation reaction occurs with a carboxylic acid, anhydride, or acyl halide (e.g., acyl chloride) (condensation reactions are usually carried out in the presence of a condensing agent, catalyst, and base; in condensation reactions, if the anhydride is highly reactive, the condensing agent can be omitted), or with a substance used to provide said R. 10 The alkyl halide undergoes a nucleophilic substitution reaction to prepare compound I-3 with an acyl or ether bond; then, under basic conditions, compound I-3 with an acyl or ether bond is reacted with a compound used to provide the R. 11 Organic amines undergo selective intermolecular Michael addition reactions to prepare guaiacol-type sesquiterpene lactone pseudo-natural product derivatives (referred to as compound V) with the structure shown in Formula V above.

[0049] Preferably, the conditions for the condensation reaction (or nucleophilic substitution reaction) in the preparation of guaiac-type sesquiterpene lactone pseudo-natural product derivatives with the structure shown in Formula V above are the same as the conditions for the condensation reaction (or nucleophilic substitution reaction) in the preparation of guaiac-type sesquiterpene lactone pseudo-natural product derivatives with the structure shown in Formula III above.

[0050] Preferably, in the preparation of guaiacol-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula V above, the base used for the selective intermolecular Michael addition reaction (providing basic conditions) is triethanolamine (TEA), diisopropylamine, or DBU, etc.

[0051] Preferably, in the preparation of guaiacol-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula V above, the selective intermolecular Michael addition reaction is carried out using methanol, tetrahydrofuran, acetonitrile, or toluene as the reaction solvent.

[0052] Preferably, in the preparation of guaiacane-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula V above, the selective intermolecular Michael addition reaction is carried out at a temperature of 25–35 °C for 12–20 h, and the compound I-3 with an acyl or ether bond reacts with the compound used to provide the R. 11 The molar ratio of the organic amines was 1:1.1 to 2. After the reaction was completed, the solvent was evaporated, and the residue was successively dissolved (in dichloromethane or ethyl acetate), washed, dried, and concentrated under reduced pressure to obtain compound V.

[0053] Sixthly, an anti-inflammatory drug is provided, the main active ingredient of which is the above-mentioned guaiacol-type sesquiterpene lactone compound (see Formula I above).

[0054] Preferably, the main active ingredient of the drug is the above-mentioned guaiacol-type sesquiterpene lactone pseudonatural product (e.g., compound I-1, compound I-2, compound I-3).

[0055] Preferably, the main active ingredient of the drug is the above-mentioned guaiacol-type sesquiterpene lactone pseudo-natural product derivative (e.g., compound II, compound III, compound V).

[0056] In a seventh aspect, a medicament is provided for the prevention and / or treatment of NLRP3 inflammasome-related diseases, wherein the main active ingredient of the medicament is the above-mentioned guaiacol-type sesquiterpene lactone compound (see Formula I above).

[0057] Preferably, the main active ingredient of the drug is the above-mentioned guaiacol-type sesquiterpene lactone pseudonatural product (e.g., compound I-1, compound I-2, compound I-3).

[0058] Preferably, the main active ingredient of the drug is the above-mentioned guaiacol-type sesquiterpene lactone pseudo-natural product derivative (e.g., compound II, compound III, compound V).

[0059] For the above-mentioned anti-inflammatory drugs and drugs used to prevent and / or treat NLRP3 inflammasome-related diseases, in addition to the main active ingredient, they also contain small amounts of minor ingredients and / or pharmaceutical excipients that do not affect the efficacy of the main active ingredient. For example, these drugs may contain sweeteners to improve taste, antioxidants to prevent oxidation, and pharmaceutically acceptable carriers.

[0060] For the above-mentioned anti-inflammatory drugs and drugs used to prevent and / or treat NLRP3 inflammasome-related diseases, there are no restrictions on the dosage form, as long as the main active ingredient can be effectively delivered into 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, etc., or in sustained-release dosage forms such as nano-formulations.

[0061] Eighthly, the application of the above-mentioned guaiacol-type sesquiterpene lactones (especially the above-mentioned guaiacol-type sesquiterpene lactone pseudo-natural products and their derivatives) in the preparation of anti-inflammatory drugs is provided.

[0062] Ninthly, the use of the above-mentioned guaiacol-type sesquiterpene lactones (especially the above-mentioned guaiacol-type sesquiterpene lactone pseudonatural products and their derivatives) in the preparation of medicaments for the prevention and / or treatment of NLRP3 inflammasome-related diseases is provided.

[0063] In a tenth aspect, the use of the above-mentioned guaiacol-type sesquiterpene lactone compounds (especially the above-mentioned guaiacol-type sesquiterpene lactone pseudonatural products and their derivatives) in the preparation of medicaments for treating nonspecific inflammatory bowel diseases (e.g., inflammatory bowel disease) or arthritis is provided.

[0064] The following are definitions of some terms that appear in the technical solutions of this invention:

[0065] The term "pharmaceutically acceptable salt" refers to the salt (e.g., sodium salt, hydrochloride salt, etc.) formed by the guaiacane-type sesquiterpene lactone compounds of this invention with a base, inorganic acid, or organic acid; the inorganic acid includes hydrofluoric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, nitric acid, and sulfuric acid; the organic acid includes formic acid, acetic acid, propionic acid, succinic acid, 1,5-naphthalenedisulfonic acid, linaloic acid, oxalic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, valeric 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, thioglycolic acid, sarcosine, glycine, and other amino acids; wherein "pharmaceutical acceptable" means suitable for human use without excessive adverse side effects (e.g., toxicity, irritation, and allergic reactions), i.e., with a reasonable benefit / risk ratio.

[0066] The term "tautomer" refers to a functional group isomer that is produced by the rapid movement of an atom in two positions within a molecule.

[0067] The term "stereoisomer" refers to isomers that are produced by different spatial arrangements of atoms in a molecule.

[0068] The term "precursor compound" refers to a compound that is inactive in vitro but can be transformed into a drug-active compound through metabolism or chemical reactions in vivo. For example, one or more of the compounds II described above are potential precursor compounds of compound I-1 described above.

[0069] The beneficial effects of this invention are reflected in:

[0070] This invention utilizes pharmacological analysis and drug activity testing of synthesized guaiacol-type sesquiterpene lactone pseudo-natural products and their derivatives with the general formula I. The results show that these compounds exhibit inhibitory effects on erythrocyte hemolysis in experiments, particularly compounds I-2 and III-12, which show significantly better inhibitory effects than the positive control drug MCC950, which specifically targets the NLRP3 inflammasome, even at lower concentrations. This invention also screens candidate drug molecules for NLRP3 inflammasome-related diseases using cellular level experiments, suggesting that guaiacol-type sesquiterpene lactone pseudo-natural products and their derivatives with the general formula I can inhibit inflammation and have the potential and medicinal value to be developed into anti-inflammatory drugs for the prevention and / or treatment of NLRP3-related inflammatory diseases such as inflammatory bowel disease. Furthermore, it provides novel parent or core structures of guaiacol-type sesquiterpene lactone compounds (e.g., compounds I-2, I-3) for subsequent large-scale drug molecule synthesis and screening, which is of great significance for broadening the application scope of guaiacol-type sesquiterpene lactone compounds. Attached Figure Description

[0071] Figure 1-1 shows the synthetic routes for intermediates (compound 2, compound 6a, and compound 6b).

[0072] Figure 1-2 shows the synthetic routes of intermediates (compound 12 and compound 20).

[0073] Figures 1-3 show the synthetic routes for guaiacane-type sesquiterpene lactone pseudonatural products and their derivatives using the general formula I structure.

[0074] Figure 2 shows the synthetic route of compound I-3.

[0075] Figure 3 shows the synthetic route of compound I-1.

[0076] Figure 4 shows the synthetic route of compound I-2.

[0077] Figure 5 shows the synthetic route of compound II-9.

[0078] Figure 6 shows the synthetic route of compound III-12.

[0079] Figure 7 shows the synthetic route of compound V-13.

[0080] Figure 8 shows the experimental results of the inhibitory activity of representative compounds I-1, I-2, I-3, II-9, III-12, and V-13 on erythrocyte hemolysis (OD505 test data after drug administration). Detailed Implementation

[0081] The present invention will be further described below with reference to the accompanying drawings and embodiments. It should be understood that the embodiments are only used to explain the present invention and are not intended to limit the scope of protection of the present invention.

[0082] 1. Design and Synthesis of Guaiacane-type Sesquiterpene Lactone Pseudo-Natural Products and Their Derivatives

[0083] Referring to Figures 1-1, 1-2, and 1-3, this invention uses (R)-Carvone (i.e., compound 1) and propynyl alcohol (i.e., compound 3a) and butynyl alcohol (i.e., compound 3b) as raw materials, and employs a designed series of synthetic routes. First, important intermediates (i.e., compounds 2, 6a, and 6b; where compounds 6a and 6b are obtained by nucleophilic substitution of propynyl alcohol and butynyl alcohol with protected hydroxyl groups in the presence of n-butyllithium, followed by electrophilic substitution with diisobutylaluminum hydride and iodoborane pinacol ester). Then, using these important intermediates (specifically, by performing tandem lactone reactions of compounds 2 and 6a, and compounds 2 and 6b, respectively, in the presence of boron trifluoride diethyl ether, followed by removal of the corresponding hydroxyl protecting groups from compounds 6a and 6b), novel 1,10-secreting guaiacol sesquiterpene lactones (see compound 8a, compound 6b) are prepared. Compound 8b was used to prepare novel guaiacol-type sesquiterpene lactones (specifically, by using compound 8a for Sven oxidation, reacting the product with vinyl magnesium bromide followed by catalytic cyclization using a Grubb second-generation catalyst, and by using compound 8b in the presence of Desmartin oxidant followed by Wittig reaction, and then catalytic cyclization using a Grubb second-generation catalyst). This led to the preparation of guaiacol-type sesquiterpene lactone pseudo-natural products (including: preparing compound I-3 from compound 20 and subsequently compound I-1, and preparing compound I-2 from compound 12) and the preparation of derivatives of these guaiacol-type sesquiterpene lactone pseudo-natural products (including: preparing compound II from compound I-1, preparing compound III from compound 12, and preparing compound V from compound I-3). The specific synthesis of compounds 12, 20, etc., is described below.

[0084] 1.1 Synthesis of Compound 2

[0085] 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.), using (R)-Carvone (i.e., compound 1) as the raw material, cyclopentane-1-carboxaldehyde compounds (specifically compound 2) were obtained through 6-step transformation. 1 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 are consistent with those reported in the literature.

[0086] 1.2 Synthesis of Compound 6a

[0087] (1) Add 110 mL of THF, 5 g of propynyl alcohol (i.e., compound 3a; 89.18 mmol, 1.0 equiv), and 15.49 g of imidazole (222.96 mmol, 2.5 equiv) to a 250 mL reaction flask containing a magnetic stirrer. Then, slowly add 28.32 mL of tert-butyldiphenylchlorosilane (TBDPSCl; 107.02 mmol, 1.2 equiv) dropwise to the reaction flask using a syringe over a period of approximately 25 minutes. Continue stirring at room temperature for 1.5 h. After confirming the complete reaction of the reactants by TLC, add 30 mL of saturated NH4Cl solution to terminate the reaction. Then, add 30 mL of water and extract with ethyl acetate (3 × 30 mL). Combine the organic phases and wash with 50 mL of saturated brine. Dry with anhydrous Na2SO4, filter, and concentrate the filtrate under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography with elution conditions of PE:EA = 50:1, and finally 21.96 g of colorless oil (i.e. compound 4a) was obtained. 1H NMR (400MHz, CDCl3) δ7.72 (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).

[0088] (2) Under a nitrogen (N2) atmosphere and at -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. Then, 35.02 mL of a 1.6 M n-butyllithium hexane solution (containing 56.03 mmol, 1.1 equiv. of n-butyllithium) was slowly added dropwise to the flask over approximately 20 minutes. The temperature was then maintained, and stirring continued for 1 hour. Next, at -78°C, 11.93 m... Methyl chloroformate (152.82 mmol, 3 equiv.) was slowly added dropwise to the round-bottom flask using a syringe, and stirring was continued for 1.0 h. The flask was then slowly brought to room temperature. After confirming complete reaction of the reactants by TLC, 30 mL of saturated NaHCO3 solution was added to terminate the reaction. The mixture was then extracted with ethyl acetate (3 × 40 mL), and the organic phases were combined and washed with 80 mL of saturated brine. The mixture was 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 under elution conditions of PE:EA = 40:1, ultimately yielding 7.64 g of a colorless oil (compound 5a). 1 H NMR (400MHz, CDCl3) δ7.67 (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).

[0089] (3) Under a nitrogen atmosphere and at 0°C, 87 mL of toluene and 7.62 mL of hexamethylphosphoric triamine (HMPA; 43.35 mmol, 2.0 equiv.) were added to a 250 mL round-bottom flask containing a magnetic stirrer and stirred for 20 min. Then, 21.67 mL of a 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 over a period of approximately 15 min. After that, 7.64 g of compound 5a (21.67 mmol, 1.0 equiv.) was added to the round-bottom flask all at once, and stirring continued. After stirring for 5 hours, 8.06 mL of iodomethylpinacol boronic acid ester (43.35 mmol, 2.0 equiv.) was added to the round-bottom flask via syringe. The mixture was stirred for 30 minutes, then transferred to room temperature and stirred overnight while monitoring by TLC. After confirming the complete reaction of the starting material, 20 mL of 1M HCl solution was added to the round-bottom flask to terminate the reaction. Then, 40 mL of water was added, followed by extraction with ethyl acetate (3 × 40 mL). The organic phases were combined and washed with 30 mL of 1M HCl solution, 50 mL of saturated NaHCO3 solution, and 50 mL of water. The mixture was 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 under elution conditions of PE:EA = 40:1, finally yielding 7.28 g of a yellow oily substance (i.e., compound 6a). 1 H NMR (400MHz, CDCl3) δ7.65(m,4H),7.39–7.30(m,6H),6.22(t,J=4.7Hz,1H),4.67(d d,J=4.8,1.5Hz,2H),3.56(s,3H),1.26(s,2H),1.21(s,6H),1.03(d,J=2.0Hz,9H).

[0090] 1.3 Synthesis of Compound 6b

[0091] (1) Unlike 1.2, 90 mL of THF, 5 g of butynol (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 using a syringe. Finally, 21 g of a colorless oily substance (compound 4b) was obtained. 1H NMR (400MHz, CDCl3) δ7.75–7.58(m,4H),7.47–7.33(m,6H),3.80(t,J=7.1Hz,2H),2.45(t,J=7.1Hz,2H),1.94(t,J=2.6Hz,1H),1.07(s,9H).

[0092] (2) Unlike 1.2, under a 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 a 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 over approximately 20 minutes. The temperature was then maintained, and stirring continued for 1 hour. Next, at -78°C, 11.38 mL of methyl chloroformate (145.87 mmol, 3 equiv.) was slowly added dropwise to the round-bottom flask using a syringe. Finally, 12.64 g of a colorless oily substance (i.e., compound 5b) was obtained. 1 H NMR (400MHz, CDCl3) δ7.69 (d, J = 6.5Hz, 4H), 7.51–7.33 (m, 6H), 4.41 (s, 2H), 3.78 (s, 3H), 1.07 (s, 9H).

[0093] (3) Unlike 1.2, under a nitrogen atmosphere and at 0°C, 87 mL of toluene and 7.67 mL of hexamethylphosphoric acid triamine (HMPA; 43.65 mmol, 2.0 equiv.) were added to a 250 mL round-bottom flask containing a magnetic stirrer and stirred for 20 min. Then, 21.83 mL of a 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 over a period of approximately 15 min. After that, 8 g of compound 5b (21.83 mmol, 1.0 equiv.) was added to the round-bottom flask all at once, and stirring was continued for 5 h. Then, 8.12 mL of iodomethylpinacol boronic acid ester (43.65 mmol, 2.0 equiv.) was added to the round-bottom flask through a syringe. Finally, 6.42 g of a yellow oily substance (i.e., compound 6b) was obtained. 1H NMR (400MHz, CDCl3) δ7.71–7.63(m,4H),7.44–7.34(m,6H),6.03(t,J=7.2Hz,1H),3.74(d,J=6.5 Hz, 2H), 3.70 (s, 3H), 2.78 (q, J = 6.8Hz, 1H), 1.85 (s, 2H), 1.28 (s, 2H), 1.23 (s, 12H), 1.05 (S, 9H).

[0094] 1.4 Synthesis of Compound 8a

[0095] (1) Under a nitrogen atmosphere and at 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 stir bar. Then, 47.85 μL of boron trifluoride diethyl ether (371.68 μmol, 0.1 equiv.) was added to the reaction flask through a microsyringe. The mixture was stirred at 0°C for 6 h, then heated to room temperature and stirred for 48 h. After confirming the complete reaction of the raw materials by TLC monitoring, 15 mL of a mixed solution of NH4Cl:NH4OH = 9:1 was added to terminate the reaction. Then, 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. The mixture was then dried with 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 with elution conditions of PE:EA = 50:1, and finally 1.51 g of pale yellow oil (i.e. compound 7a) was obtained. 1 H NMR (400MHz, CDCl3) δ7.68–7.57(m,4H),7.48–7.43(m,2H),7.42–7.36(m,4H),6.23(d,J=2.6Hz,1 H),5.51(d,J=2.2Hz,1H),4.90(s,1H),4.84(s,1H),4.48(m,1H),4.14(t,J=4.5Hz,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= 6.1Hz,9H),1.01(s,3H),0.89(s,9H),0.03(s,6H).IR(neat):3071,2954,2928,2856,1763,1651cm -1 ; 13C NMR (151MHz, CDCl3) δ170.58,144.27,136.60,135.58,132.81,132.79,129.98,127.86,127.83,122.63,112.55,81.14,77. 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.

[0096] (2) Under a 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 flask was cooled to 0 °C in an ice-water bath. Then, 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 confirming the complete reaction of the reactants by TLC, 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. The mixture was dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography with an elution ratio of PE:EA = 5:1, and 672.3 mg of a colorless oil (i.e., compound 8a) was finally obtained. 1 H NMR (400MHz, CDCl3) δ6.27(d,J=2.8Hz,1H),5.66(d,J=2.4Hz,1H),4.90(t,J=1 .8Hz,1H),4.84–4.81(m,1H),4.35(d,J=5.2Hz,1H),4.14–4.11(m,1H),3.70(d ,J=6.3Hz,2H),3.02(m,J=1H),2.97–2.90(m,1H),1.99(m,3H),1.77(d,J=1.2H z,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 (101MHz, CDCl3) δ170.45,144.33,136.38,122.81,112.40,80.90,74.52,63.85, 50.59,45.93,45.30,39.83,38.55,29.69,25.87,23.40,18.16,17.06,-4.66,-5.01.

[0097] 1.5 Synthesis of Compound 8b

[0098] (1) Unlike 1.4, under a 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 flask containing a magnetic stirrer. Then, 45.57 μL of boron trifluoride diethyl ether (353.98 μmol, 0.1 equiv.) was added to the reaction flask using a microsyringe. Finally, 991.1 mg of a pale yellow oil (i.e., compound 7b) was obtained. 1 H NMR (400MHz, CDCl3) δ7.69–7.59(m,6H),7.45-7.39(m,6H),6.18(d,J=2.6Hz,1H), 5.47(d,J=2.2Hz,1H),4.88(s,1H),4.81(s,1H),4.13(d,J=1.8Hz,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 (151MHz, CDCl3) δ170.33,143.65,138.71,135.09,132.85,129.41,127.35,121.46,112.06,82.95,76.80,76.59,76.3 8,74.10,59.93,52.98,49.76,45.46,39.36,38.88,37.92,36.13,26.44,25.44,22.98,18.69,17.72,16.62,-5.11,-5.45.

[0099] (2) Unlike 1.4: Under a 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. 1H NMR (600MHz, CDCl3) δ6.22(d,J=2.8Hz,1H),5.58(d,J=2.5Hz,1H),4.90(t,J=1.7Hz,1H),4.86–4.79(m,1H),4.21(d,J=5.5Hz,1H),4.11( t,J=4.2Hz,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 (151MHz, CDCl3) δ170.80,144.22,139.29,121.86,112.45,83.71,77.05,76.84,74. 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.

[0100] 1.6 Synthesis of Compound 12

[0101] (1) Under a nitrogen atmosphere, 40 mL of anhydrous DCM and 0.2 mL of oxaloyl chloride (2.3 mmol, 1.3 equiv.) were added to a 100 mL round-bottom flask containing a magnetic stir bar. The flask was cooled to -78 °C, and then 0.28 mL of dimethyl sulfoxide (DMSO; 3.89 mmol, 2.2 equiv.) was added dropwise to the flask over approximately 10 min. The mixture was then stirred at -78 °C for 30 min. After that, 672.3 mg of compound 8a (1.77 mmol, 1.0 equiv.) was added. The solution was added to the round-bottom flask via syringe, and the mixture was stirred at -78°C for 1 hour. Then, 1.23 mL of triethylamine (8.83 mmol, 5 equiv.) was added to the flask, and the mixture was stirred for 30 minutes. The mixture was then transferred to room temperature and stirred overnight while monitoring with TLC. After confirming complete reaction of the starting material, 20 mL of water was added to terminate the reaction. The mixture was then extracted with DCM (3 × 20 mL), and the organic phases were combined and washed with 20 mL of saturated brine. The mixture was 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 with an elution ratio of PE:EA = 5:1, ultimately yielding a white solid (compound 10). 1H NMR (400MHz, CDCl3) δ10.16(s,1H),5.26(s,1H),5.13(q,J=2.1Hz,1H),4.94(d,J=1 .9Hz,1H),4.86(s,1H),4.03(t,J=4.5Hz,1H),3.11(t,J=9.4,1H),2.45–2.39(m,1H ),2.23(d,J=2.2Hz,3H),1.90(d,J=13.0Hz,1H),1.83(s,3H),1.56(d,J=12.4Hz,1H ),1.34(t,J=7.0Hz,1H),0.83(s,9H),0.69(d,J=7.1Hz,3H),-0.03(d,J=5.7Hz,6H); 13 C NMR (101MHz, CDCl3) δ186.04,173.47,151.68,143.78,140.47,112.53,80.21,74 .33,46.98,45.43,39.41,36.56,25.82,23.56,18.11,16.06,9.21,-4.70,-5.07.

[0102] (2) Under a nitrogen atmosphere and at -78°C, 7.5 mL of anhydrous THF and 82.9 mg of compound 10 (218.97 μmol, 1.0 equiv.) were added to a 25 mL round-bottom flask containing a magnetic stir bar. Then, 0.88 mL of 1.0 M vinyl magnesium bromide THF solution (containing 875.9 μmol, 4.0 equiv. of vinyl magnesium bromide) was slowly added to the round-bottom flask over approximately 15 min. The temperature was then kept constant and the mixture was stirred for 30 min. The temperature was then raised to room temperature and stirred overnight while monitoring with TLC. After confirming that the reaction of the starting materials was complete, 10 mL of saturated NH4Cl solution was added dropwise to terminate 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. The mixture was then dried with 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 with an elution condition of PE:EA = 5:1, and 23.9 mg of colorless solid (i.e. compound 11) was finally obtained. 1H NMR(600MHz, CDCl3) δ5.94(dd,J=16.9,10.2,6.6Hz,1H),5.41(d,J=17.1Hz,1H),5.31(d,J=10.2Hz,1H),5.1 0(d,J=6.6Hz,1H),4.99(s,1H),4.94(s,1H),4.86(s,1H),4.07(t,J=4.7Hz,1H),3.07(dd,J=14.3,9.2,5.7H z,1H),2.54(dd,J=9.4,5.4Hz,1H),2.00(t,J=12.9Hz,1H),1.97–1.92(m,1H),1.82(d,J=19.1Hz,4H),1.62( s,1H),1.52(dd,J=20.9,12.6,5.9Hz,2H),1.33–1.21(m,1H),0.85(d,J=3.8Hz,12H),-0.01(d,J=3.9Hz,6H); 13 C NMR (151MHz, CDCl3) δ174.83,160.40,144.28,136.84,125.38,117.78,112.26,81.20,77.25,77.05 ,76.83,74.57,69.99,47.14,45.78,39.25,36.29,25.90,23.54,18.18,16.03,9.33,-4.66,-5.01.

[0103] (3) Under a nitrogen atmosphere, 35 mL of DCM and 73.1 mg of compound 11 (179.77 μmol, 1.0 equiv.) were added to a 100 mL round-bottom flask containing a magnetic stirrer. Then, 7.8 mg of Grubbs II catalyst (8.99 μmol, 0.05 equiv.) was added to the round-bottom flask. The mixture was transferred to an oil bath at 40 °C and heated and stirred overnight. TLC monitoring was performed to confirm that the reaction of the starting materials was complete. The solvent was then evaporated to dryness to obtain the crude product. The crude product was purified by silica gel column chromatography under elution conditions of PE:EA = 5:1, and 49.6 mg of white solid (i.e., compound 12) was finally obtained. 1H NMR(600MHz, CDCl3) δ5.79(d,J=7.5Hz,1H),4.95(d,J=7.3Hz,1H),4.60(d,J=11 .5Hz,1H),4.06(t,J=4.0Hz,1H),3.47(q,J=8.5Hz,1H),2.09(m,1H),2.05–1.99 (m,1H),1.97–1.93(m,1H),1.92(d,J=1.8Hz,3H),1.81–1.77(m,3H),1.75(m,1H ),1.27–1.21(m,1H),1.04(d,J=6.7Hz,3H),0.85(s,9H),0.02(d,J=3.0Hz,6H); 13 C NMR (151MHz, CDCl3) δ174.31,161.34,144.54,125.84,122.08,83.51,83.51,83.36,77.22,77.00, 76.79,74.21,61.57,46.87,45.00,41.81,40.05,25.79,25.71,18.09,15.34,8.66,-4.66,-4.93.

[0104] 1.7 Synthesis of Compound 20

[0105] (1) Under nitrogen atmosphere and at 0℃, 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. Then, under stirring, 485.7 mg of Dess-Martin oxidant (1.09 mmol, 1.30 equiv.) was added to the round-bottom flask. The temperature was raised to room temperature and stirring was continued for 1 h. After confirming that the reaction of the raw materials was complete by TLC monitoring, 15 mL of saturated Na2S2O3 solution was added to quench the reaction. Then, the reaction was extracted with dichloromethane (3 × 15 mL). The organic phases were combined and washed successively with 20 mL of saturated NaHCO3 solution and 20 mL of saturated brine. The mixture was then dried with 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 with an elution condition of PE:EA = 5:1, and 239 mg of white solid (i.e. compound 16) was finally obtained. 1H NMR (600MHz, CDCl3) δ9.79 (s, 1H), 6.23 (d, J = 2.8Hz, 1H), 5.56 (d, J = 2.4Hz, 1H), 4.88(t,J=1.8Hz,1H),4.81(s,1H),4.12(t,J=4.9Hz,1H),4.10(m,1H),3.29(d, J=6.7Hz,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=7.0Hz,3H),0.85(s,9H),0.00(s,6H); 13 C NMR (151MHz, CDCl3) δ198.89,169.97,144.19,138.34,122.72,112.40,83.11,77.25,77.04, 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.

[0106] (2) Under a nitrogen atmosphere, 25 mL of THF and 1.33 g of methyltriphenylphosphine bromide (3.65 mmol, 6.0 equiv.) were added to a 50 mL reaction flask containing a magnetic stirrer. The mixture was cooled to -50 °C. Then, under stirring, 1.5 mL of a 2.5 M n-butyllithium hexane solution (containing 3.65 mmol, 6.0 equiv. of n-butyllithium) was added dropwise to the reaction flask. The mixture was stirred at -50 °C for 1 h. Subsequently, 239 mg of a compound dissolved in 5.0 mL of anhydrous THF was added. Compound 16 (0.61 mmol, 1.0 equiv.) was added dropwise to the reaction flask over approximately 8 minutes. The mixture was stirred at -50°C for 30 minutes, then transferred to room temperature and stirred for 3 hours. After confirming complete reaction of the starting materials by TLC, 10 mL of saturated NH4Cl solution was added to quench the reaction. The mixture was then extracted with ethyl acetate (3 × 10 mL). The combined organic phases were washed with 20 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated to dryness to obtain the crude product. The crude product was purified by silica gel column chromatography at a PE:EA ratio of 10:1, ultimately yielding 153.1 mg of a colorless oil (i.e., compound 17). 1H NMR (600MHz, CDCl3) δ6.22(d,J=2.9Hz,1H),5.72(m,1H),5.57(d,J=2.5Hz,1H),5.16–5.05( m,2H),4.89(t,J=1.8Hz,1H),4.82(s,1H),4.16(d,J=6.1Hz,1H),4.12(t,J=4.8Hz,1H),2.9 9(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=5.7Hz,2H),1.01(d,J=7.0Hz,3H),0.85(s,9H),0.00(s,6H); 13 C NMR (151MHz, 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.

[0107] (3) Under a nitrogen atmosphere, 17 mL of anhydrous DCM and 33.5 mg of compound 17 (85.76 μmol, 1.0 equiv.) were added to a 50 mL round-bottom flask containing a magnetic stirrer. Then, under stirring, 3.7 mg of Grubbs II catalyst (4.29 μmol, 0.05 equiv.) was added to the round-bottom flask. At this time, the solution was wine-red in color. The mixture was then heated and stirred in an oil bath at 40 °C for 12 h. After confirming the complete reaction of the reactants by TLC, the solvent was evaporated to dryness to obtain the crude product. The crude product was purified by silica gel column chromatography with elution conditions of PE:EA = 20:1, and finally 28.9 mg of white solid (i.e., compound 20) was obtained. 1H NMR (600MHz, CDCl3) δ6.17(d,J=3.5Hz,1H),5.45(d,J=3.1Hz,1H),5.44–5.36(m,1H),4.13–4.03( m,1H),3.89(t,J=11.2Hz,1H),3.10(dd,J=13.9,7.3Hz,1H),2.67(d,J=12.1Hz,1H),2.61–2.53(m ,1H),2.20(t,J=7.0Hz,1H),2.02(t,J=12.7Hz,1H),1.96(d,J=3.0Hz,1H),1.90–1.83(m,1H),1.7 7–1.71(m,3H),1.51(t,J=12.7Hz,1H),1.07(d,J=7.0Hz,3H),0.87(s,9H),0.03(d,J=4.2Hz,6H); 13 C NMR (151MHz, CDCl3) δ170.35,140.22,137.00,120.04,119.85,87.32,77.26,77.05,76.83, 73.57,52.18,45.26,44.66,42.65,42.26,30.62,28.44,25.82,18.15,15.75,-4.67,-4.91.

[0108] 2. Examples of Synthesis of Guaiacane-type Sesquiterpene Lactone Pseudo-Natural Products and Their Derivatives

[0109] 2.1 Synthesis of Compound I-3

[0110] Referring to Figure 2, under a nitrogen atmosphere and at 0°C, 2.5 mL of anhydrous THF and 25.2 mg of compound 20 (69.5 μmol, 1.0 equiv.) were added dropwise to a 10 mL reaction flask containing a magnetic stirrer. Then, under stirring, 0.1 mL of TBAF (104.25 μmol, 1.5 equiv.) was added dropwise to the reaction flask. After the addition was complete, the flask was transferred to room temperature and stirred for 2 h. After confirming the complete reaction of the reactants by TLC, 10 mL of water was added to quench the reaction. The reaction system 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 (0.08 MPa, 35°C) to obtain the crude product. The crude product was purified by silica gel column chromatography with an elution ratio of PE:EA = 5:1. The collected eluent was dried under vacuum to obtain 12 mg of a white solid, namely compound I-3. The specific synthesis results and compound identification of compound I-3 are as follows.

[0111] Yield 70%. HPLC purity (98%). f =0.3(PE:EA=2:1). mp:144.9~155.2℃. 1 H NMR (600MHz, CDCl3) δ6.18(d,J=3.5Hz,1H),5.45(d,J=14.6Hz,2H),4.17(t,J=4.9Hz,1H),3.92(t,J=11.1Hz,1H),3.11(m,1H),2 .70(m,1H),2.59(m,1H),2.28(m,1H),2.15(d,J=13.2Hz,1H),1.95(m,2H),1.77(s,3H),1.72–1.55(m,1H),1.16(d,J=7.1Hz,3H). 13 CNMR (151MHz, CDCl3) δ170.26,140.07,136.61,120.28,120.07,86.99,77.26, 77.05,76.84,73.27,52.36,44.60,44.49,42.63,41.48,30.62,28.42,14.84. IR(neat):3416,2924,1750,1641cm -1 HRMS(ESI)m / z:[M+H] + Calculated for (using the molecular formula below as a reference) C 15 H 20 O3249.1485; found (actual measured value) 249.1489.

[0112] 2.2 Synthesis of Compound I-1

[0113] Referring to Figure 3, at 0°C, 4 mL of anhydrous dichloromethane and 18.6 mg of compound I-3 (74.9 mmol, 1.00 equiv.) were added to a 20 mL reaction flask containing a magnetic stir bar. Then, under stirring, 41.3 mg of Dess-Martin oxidant (DMP; 97.37 mmol, 1.30 equiv.) was added to the reaction flask. The temperature was then raised to room temperature, and stirring was continued for 2 h. After confirming the complete reaction of the reactants by TLC, 10 mL of saturated Na2S2O3 solution was added to quench the reaction. The reaction system was then extracted with DCM (3 × 10 mL). The extracted organic phases were combined, and the mixed organic phases were washed successively with 10 mL of saturated NaHCO3 solution and 10 mL of saturated brine. The mixture was then dried with anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure (0.08 MPa, 35°C) to obtain the crude product. The crude product was purified by silica gel column chromatography with an elution ratio of PE:EA = 5:1. The collected eluent was dried under vacuum to obtain 16.7 mg of a white solid, namely compound I-1. The specific synthesis results and compound identification of compound I-1 are as follows.

[0114] Yield 96%. HPLC purity (96%). f =0.25(PE:EA=5:1). mp:133.2~135.8℃. 1 H NMR (600MHz, CDCl3) δ6.21(d,J=3.4Hz,1H),5.51(d,J=10.1Hz,2H),4.05(t,J=9.9Hz,1H),3.21(q,J=7.9Hz,1H),2.96(m,1H),2.74(m,1H),2.54( t,J=7.0Hz,1H),2.50(d,J=8.1Hz,1H),2.33(m,1H),2.24(dt,J=11.8,5. 9Hz, 1H), 2.11–2.05 (m, 1H), 1.77 (d, J = 2.2Hz, 3H), 1.22 (d, J = 7.5Hz, 3H). 13 C NMR (151MHz, CDCl3) δ218.34,169.68,139.77,135.51,122.11,120.38,84.10,77. 25,77.04,76.83,52.45,47.60,47.49,43.86,38.38,29.78,29.71,27.13,15.35. IR(neat):3355,2970,2930,2881,1754,1732,1661cm -1 HRMS(ESI)m / z:[M+H] +Calculated for C 15 H 18 O3 247.1328; found 247.1342.

[0115] 2.3 Synthesis of Compound I-2

[0116] Referring to Figure 4, at 0°C, 3.5 mL of anhydrous THF and 26.5 mg of compound 12 (70 μmol, 1.0 equiv) were added to a 10 mL PTFE plastic reaction flask containing a magnetic stirrer. Then, under stirring, 100 μL of 70% pyridine hydrofluoric acid was added to the reaction flask. The mixture was then stirred overnight at 0°C and monitored by TLC. After confirming complete reaction of the starting materials, 5 mL of water was added dropwise to the reaction flask to terminate the reaction. Another 5 mL of water was then added dropwise to the reaction flask. The resulting mixture was extracted with ethyl acetate (3 × 10 mL). The combined organic layers were washed with 10 mL of saturated brine, dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure (0.08 MPa, 35°C) to obtain the crude product. The crude product was purified by silica gel column chromatography with an elution ratio of PE:EA = 2:1. The collected eluent was dried under vacuum to obtain 8.5 mg of a white solid, namely compound I-2. The specific synthesis results and compound identification of compound I-2 are as follows.

[0117] Yield 46%. HPLC purity (99%). f =0.3(PE:EA=2:1). mp:95.3~98.7℃. 1 H NMR (600MHz, CDCl3) δ6.22(d,J=11.8Hz,1H),6.03(d,J=11.8Hz,1H),4.82(d,J=11.9,1H),4.18(t,J=4.0Hz,1H),2.63(t,J=16.4H z,2H),2.56(d,J=18.3Hz,1H),2.23(m,1H),1.88(d,J=1.7Hz,3H),1.82(d,J=2.2Hz,3H),1.59(d,J=7.0Hz,1H),1.28–1.20(m,5H). 13CNMR (151MHz, CDCl3) δ174.26,156.38,142.90,137.48,127.16,121.21,116.5 8,82.57,77.24,77.03,76.81,74.50,49.03,44.87,41.75,21.81,14.44,8.46. IR(neat):3488,2958,2920,2886,1723,1645,1618cm -1 HRMS(ESI)m / z:[M+H] + Calculated for C 15 H 20 O4 265.1362; found 265.1367.

[0118] 2.4 Synthesis of Compound II-9

[0119] Referring to Figure 5, under a nitrogen atmosphere, compound I-1 (44.3 mg, 0.2 mmol) and 5-fluorouracil (28.6 mg, 0.22 mmol) were added to a 25 mL double-necked flask, followed by K₂CO₃ (55.3 mg, 0.4 mmol), and then 8 mL of THF. The mixture was stirred overnight at room temperature. The reaction was monitored by TLC until completion. The mixture was 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 petroleum ether:ethyl acetate at a 1:1 elution ratio to obtain 72.5 mg of a colorless solid, with a yield of 96.3%. This was identified as compound II-9, HRMS (ESI) m / z: [M+H]. + Calculated for C 19 H 21 FN2O5 377.1434; found 377.1439.

[0120] For stability and ease of storage, the obtained compound II-9 was converted to its sodium salt using the following steps: Compound II-9 (37.6 mg, 0.1 mmol) and 5 mL of methanol were added to a 25 mL double-necked flask. Then, under stirring, 1 mL of 0.1 M NaOH solution was added to the flask. The reaction was stopped after stirring for 30 min. Extraction was performed using DCM (3 × 3 mL). The separated aqueous phase was freeze-dried to obtain 35.73 mg of a white powdery solid, with a yield of 89.7%. It was identified as the sodium salt of compound II-9 (abbreviated as II-9-Na), HRMS(ESI) m / z: [M-Na]. - Calculated for C 19 H 20FN2O5Na375.1356; found 375.1351.

[0121] 2.5 Synthesis of Compound III-12

[0122] Referring to Figure 6, under a nitrogen atmosphere and at 0 °C, compound 12 (37.9 mg, 0.1 mmol), Ac₂O (12.3 mg, 0.12 mmol, 11.24 μL), DMAP (9.8 mg, 0.08 mmol), and 1 mL of pyridine were added to a 25 mL double-necked flask. The mixture was stirred for 30 min, then placed at room temperature and stirred under a nitrogen atmosphere for 12 h. After confirming the complete reaction of the starting materials by TLC, 10 drops of water were added to the reaction mixture, and stirring was continued for 30 min to quench the reaction. Then, 20 mL of ethyl acetate was added, and the mixture was dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure (0.08 MPa, 35 °C) to obtain the crude product. The crude product was purified by silica gel column chromatography using petroleum ether (60–90 °C):ethyl acetate = 10:1 as the elution conditions, yielding 39.2 mg of a colorless oily substance, with a yield of 93.2%. Identified as compound 3', HRMS(ESI) m / z: [M+H] + Calculated for C 23 H 36 O5Si 421.2332; found 421.2337.

[0123] Referring to Figure 6, under a nitrogen atmosphere and at 0°C, 3.5 mL of anhydrous THF and compound 3' (21.5 mg, 0.05 mmol) were added to a 10 mL polytetrafluoroethylene reaction flask containing a magnetic stirrer. Then, under stirring, 100 μL of 70% pyridine hydrofluoric acid was added to the reaction flask. The mixture was stirred overnight at 0°C and monitored by TLC (PE:EA = 5:1). After confirming complete reaction of the starting materials, 5 mL of water was added dropwise to the reaction flask to terminate the reaction. Then, 5 mL of water was added dropwise to the reaction flask. The resulting mixture was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with 10 mL of saturated sodium bicarbonate solution, dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure (0.08 MPa, 35°C) to obtain the crude product. The crude product was purified by silica gel column chromatography under elution conditions of PE:EA = 5:1 to obtain 13.3 mg of white solid, with a yield of 86.9%. Identified as compound III-12, HRMS(ESI) m / z: [M+H] + Calculated for C 17 H 22 O5 307.1467; found 307.1475.

[0124] 2.6 Synthesis of Compound V-13

[0125] Referring to Figure 7, under a nitrogen atmosphere and at 0°C, compound I-3 (24.8 mg, 0.1 mmol), propionic anhydride (15.6 mg, 0.12 mmol, 154 μL), DMAP (9.8 mg, 0.08 mmol), and 1 mL of pyridine were added to a 25 mL double-necked flask. The mixture was stirred for 30 min, then placed at room temperature and stirred under a nitrogen atmosphere for 12 h. After confirming the complete reaction of the starting materials by TLC, 10 drops of water were added to the reaction mixture, and stirring was continued for 30 min to quench the reaction. Then, 20 mL of ethyl acetate was added, and the mixture was dried over anhydrous Na2SO4. After filtration, 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 petroleum ether (60–90°C):ethyl acetate = 5:1 as the elution conditions, yielding 29.7 mg of a white solid, with a yield of 97.6%. Identified as compound 4', HRMS(ESI) m / z: [M+H] + Calculated for C 18 H 24 O4 305.1675; found 305.1682.

[0126] Referring to Figure 7, dimethylamine hydrochloride (48.9 mg, 0.6 mmol) and 10 mL of methanol were placed in a 50 mL double-necked flask. After stirring at room temperature, TEA (0.8 mmol, 112 μL) was added dropwise to the flask using a syringe. After the addition was complete, the mixture was stirred vigorously for 30 min under nitrogen protection. Then, compound 4' (152.2 mg, 0.5 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until it ended. Methanol was removed under reduced pressure (0.08 MPa, 35 °C). The residue was dissolved in 15 mL of dichloromethane and washed successively with 5 mL of citric acid, water, and saturated saline solution. The residue was then dried over anhydrous Na2SO4 and concentrated under reduced pressure (0.08 MPa, 35 °C) to give 135.1 mg of white solid (i.e., compound V-13), with a yield of 77.3%.

[0127] For stability and ease of storage, the obtained compound V-13 was salted as follows: At room temperature, compound V-13 (104.8 mg, 0.3 mmol) and 5 mL of dichloromethane were added to a 25 mL round-bottom flask and stirred. After compound V-13 was completely dissolved, 0.1 M HCl solution was slowly added dropwise until the pH of the solution reached approximately 3. Then, 5 mL of purified water was added to the round-bottom flask, and after stirring for 5 min, the organic phase was separated using a separatory funnel. The aqueous phase was extracted using DCM (3 × 5 mL), and the extracted aqueous phase was freeze-dried to obtain 102.1 mg of a white powdery solid, with a yield of 88.2%. This was identified as compound V-13 hydrochloride (abbreviated as V-13-HCl), HRMS (ESI) m / z: [M-Cl]. + Calculated for C 20 H 32 ClNO4 350.2331; found 350.2337.

[0128] 3. Preparation of tablets containing guaiacane-type sesquiterpene lactone pseudonatural products and their derivatives

[0129] 500 mg of compounds I-1, I-2, I-3, II-9-Na, III-12 or V-13-HCl were mixed with 4.275 g of excipients (white peony: lactose = 7:3, mass ratio), 95% ethanol was added, granulation was performed, drying was carried out, and the granules were sieved. 0.125 g of sodium stearate was added, and the mixture was mixed evenly and then compressed into tablets (each tablet weighing 200 mg) to obtain tablets containing 10 mg of compounds I-1, I-2, I-3, II-9-Na, III-12 or V-13-HCl.

[0130] 4. Preparation of guaiacane-type sesquiterpene lactone pseudonatural products and their derivatives as powder injections

[0131] 100 mg of compound I-1, I-2, I-3, II-9-Na, III-12 or V-13-HCl was dissolved in 17 mL of water for injection with 500 mg of mannitol. After initial mixing, the volume was adjusted to 20 mL. The resulting solution was filtered, placed into vials (1 mL per vial), lyophilized, sealed, and sterilized to obtain lyophilized powder injections containing 5 mg of compound I-1, I-2, I-3, II-9-Na, III-12 or V-13-HCl per vial.

[0132] 5. Preparation of capsules containing guaiacane-type sesquiterpene lactone pseudo-natural products and their derivatives

[0133] 150 mg of compounds I-1, I-2, I-3, II-9-Na, III-12 or V-13-HCl were mixed with 1.35 g of excipients (white peony: lactose = 7:3, mass ratio), 95% ethanol was added, granulated, dried, sized (sieved), and filled into capsules (each capsule weighing 150 mg) to obtain capsules containing 15 mg of compounds I-1, I-2, I-3, II-9-Na, III-12 or V-13-HCl.

[0134] 6. Assay for detecting the inhibitory activity of compounds I-1, I-2, I-3, II-9, III-12, and V-13 on erythrocyte hemolysis.

[0135] 6.1 Experimental Materials

[0136] (1) Experimental reagents: Plasma free hemoglobin assay kit (Rilda).

[0137] (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 anticoagulant, centrifuged, and two layers were obtained, the upper layer being plasma and the lower layer being red blood cells. The lower layer of red blood cells was used for the experiment.

[0138] (3) Experimental instruments: enzyme-linked immunosorbent assay (ELISA) reader, etc.

[0139] 6.2 Experimental Scheme

[0140] In the erythrocyte hemolysis model, hemolysis refers to the rupture or destruction of erythrocytes (RBCs), resulting in the release of hemoglobin and other cellular contents into the surrounding environment.

[0141] Small molecule compounds at different concentrations (0 μM, 2 μM, 4 μM, 8 μM, 16 μM) were co-incubated with erythrocytes at 37°C for 30 min. The small molecule compounds were I-1, I-2, I-3, II-9, III-12, V-13, or MCC950 (as a positive control). Subsequently, nigericin at a final concentration of 1 μM was added 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 levels were quantified using a kit. The activity of the small molecule compounds was tested based on the direct proportionality between hemoglobin content and absorbance value (OD505).

[0142] Table 1. Experimental Groups and Treatments

[0143] Note: The total volume of each group in Table 1 is 200 μL. N represents Nigericin, and the final concentration of Nigericin in the system is 1 μM. X represents the volume of drug solution of I-1, I-2, I-3, II-9, III-12, V-13 or MCC950 (the solvent for preparing the drug solution is 0.9% physiological saline). NS represents 0.9% physiological saline.

[0144] 6.3 Specific Experimental Procedures

[0145] S1 adds 1 mL of physiological saline (0.9% NaCl) to a 1.5 mL EP tube.

[0146] S2 aspirates 100 μL of lower red blood cells (i.e., red blood cells separated from blood) and adds them to an EP tube containing 1.5 mL of physiological saline.

[0147] S3 was centrifuged at 3000 r / min for 5 min.

[0148] After centrifugation at S4, discard 1 mL of supernatant, add 1 mL of physiological saline, and centrifuge again at 3000 r / min for 5 min.

[0149] After centrifugation at S5, discard the supernatant; the remaining cells are the required red blood cells (this completes the washing of red blood cells).

[0150] S6 prepares suspensions of physiological saline and red blood cells according to the 7 regimens in Table 1.

[0151] S7. The small molecule compound to be tested was added to the corresponding group of red blood cell suspensions at concentrations of 0 μL, 32 μL, 0 μL, 4 μL, 8 μL, 16 μL, and 32 μL (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 incubated at 37 °C for 30 min.

[0152] After 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.

[0153] After S9 incubation is complete, centrifuge at 2000 r / min for 3 min.

[0154] S10 aspirates the supernatant and sets it aside for later use.

[0155] S11 uses a plasma free hemoglobin assay kit to detect hemoglobin in the supernatant (detection conditions: react at 37°C for 20 min after adding reagents).

[0156] After the S12 reaction was completed, the absorbance value at a wavelength of 505 nm was measured using an ELISA reader.

[0157] 6.4 Experimental Results

[0158] As shown in Figure 8, the experimental results indicate that, using the release of hemoglobin from erythrocytes stimulated with Nigericin after 30 min of co-incubation with MCC950 as a reference, compounds I-2, I-3, III-12, and V-13 all exhibited significant inhibitory effects on erythrocyte hemolysis induced by Nigericin stimulation at treatment concentrations of 2 μM and 4 μM, and their inhibitory effects were significantly better than those of the positive control drug MCC950. Compounds I-2 and III-12 showed the best inhibitory effects at a treatment concentration of 4 μM. These results suggest that guaiacol-type sesquiterpene lactones (or guaiacol-type sesquiterpene lactone pseudo-natural products), represented by compounds I-2 and I-3, and their derivatives, have a certain inhibitory effect on erythrocyte hemolysis and could serve as candidate drug molecules for the prevention and / or treatment of NLRP3 inflammasome-related diseases.

[0159] The results of the parallel experiments in the Control group and the N+0μM group in Figure 8 fluctuated (OD505 values ​​were high and low), which may be due to the difference in red blood cells used when conducting experiments on different small molecule compounds.

[0160] In summary, based on the synthesis of guaiacol-type sesquiterpene lactone pseudo-natural products and their derivatives, this invention has discovered novel guaiacol-type sesquiterpene 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 their use as active ingredients in the preparation of anti-inflammatory drugs).

Claims

1. An ent -kaurene diterpene lactone compound, characterized in that: The compound is any one of the following: a compound having the structure shown in Formula I, or a pharmaceutically acceptable salt, tautomer, stereoisomer, or precursor compound of the compound: In Formula I: The double bond is formed by R1 and R2, and R9 is hydrogen; or the double bond is formed by R9 and R2, and R1 is methyl; or both R9 and R1 are hydrogen, and R2 is selected from any one of alkylaminomethyl, arylaminomethyl, heteroarylaminomethyl, or groups with structures as shown in Formula X: In formula X, R3, R4, and N atoms form a ring structure, which is a heterocycle containing one N atom. R5 is hydrogen; R6 is a methyl group; R7 is a carbonyl oxygen atom, -ORc, -OCOORc, -OCORc, or -OCONRcRd; R8 can be hydrogen, -ORc, -OCOORc, -OCORc, or -OCONRcRd; 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, respectively.

2. A guaiacane-type sesquiterpene lactone pseudonatural product, characterized in that: The pseudo-natural product is any one of the following: a compound having the structure shown in Formula I-1, Formula I-2 or Formula I-3, or a pharmaceutically acceptable salt, tautomer, stereoisomer or precursor compound of the compound:

3. A guaiacane-type sesquiterpene lactone pseudo-natural product derivative, characterized in that: The pseudo-natural product derivative is any one of the following: a compound having the structure shown in Formula II, Formula III or Formula V, or a pharmaceutically acceptable salt, tautomer, stereoisomer or precursor compound of the compound: Among them, R 10 The Rc is selected from any one of Rc, COORc, CORc, and CONRcRd, wherein Rc and Rd are respectively selected from any one of alkynyl, cycloalkyl, halogen, alkyl, aryl, halogen-substituted alkyl, halogen-substituted aryl, and nitro-substituted aryl; R 11 It is selected from any one of alkylamino, aromaticamino, heteroaromaticamino, or heterocyclic amino containing one N atom.

4. The guaiacol-type sesquiterpene lactone pseudo-natural product derivative according to claim 3, characterized in that: The pseudo-natural product derivative is any one of the following compounds having the structure shown in formulas II-1 to II-12, III-1 to III-12, or V-1 to V-31, or a pharmaceutically acceptable salt, tautomer, stereoisomer, or precursor compound of the compound:

5. A method for preparing a guaiacane-type sesquiterpene lactone compound, characterized in that: Includes the following steps: Using levamisole and butynol as raw materials, and through the synthesis of 1,10-secreting guaiacol sesquiterpene lactones, oxidation of the corresponding alcohol hydroxyl groups in the structure of the compound to aldehyde groups, and conversion of the generated aldehyde groups to alkenes followed by catalytic cyclization, guaiacol sesquiterpene lactone pseudo-natural products with structures as shown in Formula I-3 were prepared. Using guaiacane-type sesquiterpene lactone pseudo-natural products with structures as shown in Formula I-3, and through oxidation reactions, guaiacane-type sesquiterpene lactone pseudo-natural products with structures as shown in Formula I-3 were prepared as follows: Alternatively, using levamisole and propynyl alcohol as raw materials, and through the synthesis of 1,10-secreting guaiacol sesquiterpene lactones, oxidation of the corresponding alcohol hydroxyl groups in the structure of the compound to aldehyde groups, and conversion of the generated aldehyde groups to alkenyl groups followed by catalytic cyclization, guaiacol sesquiterpene lactone pseudo-natural products with structures as shown in Formula I-2 can be prepared.

6. A method for preparing a guaiacane-type sesquiterpene lactone compound, characterized in that: Includes the following steps: 1) Using L-carvone as the starting material, prepare guaiacol-type sesquiterpene lactone intermediates; 2) Using the guaiacol-type sesquiterpene lactone intermediate obtained in step 1, and through reactions to achieve esterification, etherification, and / or intermolecular Michael addition reactions, prepare guaiacol-type sesquiterpene lactone pseudo-natural product derivatives with structures as shown in Formula II, Formula III, or Formula V: Among them, R 10 The Rc is selected from any one of Rc, COORc, CORc, and CONRcRd, wherein Rc and Rd are respectively selected from any one of alkynyl, cycloalkyl, halogen, alkyl, aryl, halogen-substituted alkyl, halogen-substituted aryl, and nitro-substituted aryl; R 11 It is selected from any one of alkylamino, aromaticamino, heteroaromaticamino, or heterocyclic amino groups containing one nitrogen atom.

7. An anti-inflammatory drug, characterized in that: The drug includes guaiacol-type sesquiterpene lactones as described in claim 1.

8. A medicament for preventing and / or treating an NLRP3 inflammasome-related disease, characterized by: The drug includes guaiacol-type sesquiterpene lactones as described in claim 1.

9. The use of the guaiacol-type sesquiterpene lactone compound as described in claim 1 in the preparation of anti-inflammatory drugs.

10. The use of the guaiacol-type sesquiterpene lactone compound as described in claim 1 in the preparation of a medicament for the prevention and / or treatment of NLRP3 inflammasome-related diseases.

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