Guaiane-based sesquiterpene derivatives and uses thereof

Abstract

The present invention discloses a guaiane-based sesquiterpene derivative and its pharmaceutical use, and the guaiane-based sesquiterpene derivative or its pharma- ceutical acceptable salt is represented by general formula I. In this application, by using natural ingredients rich in parthenolide, dehydrocostus lactone and the like as raw materials and optimizing the structure, a novel guaiane-based sesquiterpene derivative with strong inhibitory activity against NLPR3 inflammasome activation and significantly improved chemical stability, water solubility and oral bioavailability has been discovered, and it has been experimentally verified that this derivative has an inhibitory effect against NLRP3 inflammasome activity and may be used for medicinal purposes. JPEG2024516840000060.jpg37166

Description

[Technical Field]

[0001] The present invention relates to compound derivatives and their uses, and more particularly to guaiane-based sesquiterpene derivatives and their pharmaceutical uses. [Background technology]

[0002] Inflammasomes are protein complexes that can distinguish between pathogen-associated mode molecules (PAMPs) and damage-associated mode molecules (DAMPs) in resident immune cells, such as macrophages, monocytes, and dendritic cells [Front Immunol, 2019,10: 2538.] Different types of inflammasomes, such as NLRP1, NLRP3, NLRC4, Pyrin, NLRP6, and AIM2, mediate inflammatory responses, promote the release of inflammatory cytokines, transmit signals to the immune system, initiate inflammation, and act as a bridge between innate and adaptive immunity [Cell, 2016, 165:792-800.] Unlike other types of inflammasomes, which specifically recognize DAMPs or PAMPs, the NLRP3 inflammasome can broadly recognize DAMPs and PAMPs of different origins. Therefore, research on the NLRP3 inflammasome has attracted attention from various fields. It is currently the most extensively studied inflammasome and has been demonstrated to be involved in the onset and progression of many chronic inflammatory diseases [Nat Rev Drug Discov, 2018, 17:588-606.]

[0003] The NLRP3 inflammasome consists of three parts: the receptor protein NLRP3, the regulatory protein ASC, and the effector protein pro-Caspase-1 [Immunol Rev,2015, 265:35-52.] NLRP3 inflammasome activation is divided into two steps. In step 1, the TLR4 receptor recognizes primary signals, such as PAMPs, DAMPs, or exogenous stress molecules, and activates the NF-κB pathway to upregulate the expression of NLRP3, pro-IL-1β, and pro-IL-18. In step 2, the receptor protein NLRP3 recognizes secondary signals, such as PAMPs, DAMPs, or intracellular stress molecules, and activates pro-caspase-1 by binding the adaptor protein ASC, which then cleaves and activates pro-IL-1β and pro-IL-18, promoting their maturation and secretion [Int J Mol Sci, 2019, 20:3328.] IL-1β further activates the NF-κB signaling pathway through autocrine and paracrine pathways, promoting the secretion of cytokines such as IL-1β, TNF-α, IL-6, and IL-8, inducing an inflammatory cascade and leading to chronic progression [Front Immunol, 2019, 10:276.]

[0004] Hyperactivation of the NLRP3 inflammasome is closely associated with the onset and exacerbation of various diseases, including immune disorders, autoimmune diseases, malignant tumors, skin diseases, cardiovascular diseases, liver-related diseases, renal system-related diseases, gastrointestinal diseases, central nervous system diseases, metabolic diseases, endocrine-related diseases, respiratory diseases, lymphatic system diseases, inflammation, infectious diseases, eye diseases, psychiatric disorders, and pain [Nat Med, 2015, 21:248-255; J Clin Invest, 2020, 130:1961-1976; Cell Metab, 2020, 31:580-591; Circ Res, 2018, 122:1722-1740; J Hepatol, 2017, 66:1037-1046; Ageing Res Rev, 2020, 64:101-192; Autophagy, 2019, 15:1860-1881; Brain,2020, 143:1414-1430; Mucosal Immunol, 2019, 12:1150-1163; J Clin Invest, 2018, 128:1793-1806; Immunology, 2020, 160:78-89; J Inflamm (Lond), 2015, 12:41; Nat Commun, 2020, 11:4243; Front Immunol, 2020, 11:570251; BiochemBiophys Res Commun, 2016, 477:329-335; Pharmaceutics, 2020, 12:867; Arthritis Rheumatol, 2020, 72:1192-1202; Food Chem Toxicol, 2020, 144:111588; EMBO Rep, 2020, 21:e49666; Int Immunopharmacol, 2020, 81:106257; Cells, 2019, 8: 1389; Cell Prolif, 2021, 54: e12973.] Therefore, inhibiting activation of the NLRP3 inflammasome can prevent and / or treat the above-mentioned diseases.

[0005] The structural formula of Arglabin is shown below. [ka]

[0006] The Abderrazak A research team found that arglabin, a guaiane-based sesquiterpene lactone, has extremely strong inhibitory activity against NLPR3 inflammasome activation (EC 50 =10 nM), arglabin was found to reduce NLPR3 inflammasome-associated inflammation, protect pancreatic beta cells, and prevent type 2 diabetes [Circulation, 2015, 131:1061-1070; J Pharmacol Exp Ther, 2016, 357:487-494.] Arglabin is derived from the Kazakh plant Artemisia princeps (Mugwort) and has a relatively low content of approximately 0.27% [J Nat Prop, 1999, 62: 1068-1071.] Its aqueous solubility is only 7.9 μg / mL, its chemical stability in gastric fluid is poor, its degradation rate reaches 50% within 8 hours, and its oral bioavailability is only 5%. These pharmaceutical limitations limit its clinical application. Therefore, further improvements in the chemical stability, water solubility, oral bioavailability, and resource economy of this compound are desirable. Summary of the Invention [Problem to be solved by the invention]

[0007] The object of the present invention is to provide guaiane-based sesquiterpene derivatives to improve the chemical stability, water solubility, and oral bioavailability of the compounds.Another object of the present invention is to provide a use of the compounds in the manufacture of drugs for treating NLRP3 inflammasome-related diseases. [Means for solving the problem]

[0008] The technical solution is as follows: A guaiane-based sesquiterpene derivative represented by general formula I or a pharmaceutically acceptable salt thereof: [ka] R1 and R2 together form a double bond, or R1 is hydrogen or deuterium and R2 is [ka] wherein R3 and R4 are each hydrogen, an alkyl group, or a cycloalkyl group, and R3, R4, and the N atom together form a 3- to 9-membered ring structure, which may be substituted with one or more substituents including an alkyl group, an ester group, an aryl group, an alkylaryl group, an arylalkyl group, an arylalkenyl group, an arylalkynyl group, or a heterocyclic group; R5 and R6 may be bonded to each other by a single bond to form a cyclopropane; when R5 and R6 do not form a cyclopropane, R5 is a methyl group, and R6 is a hydroxyl group, an alkoxy group, an ester group, or a halogen atom, etc., where, when R6 is not a hydroxyl group, an alkoxy group, an ester group, or a halogen atom, it can form a double bond with the carbon atom at the ortho position; R7 is hydrogen or a hydroxyl group; R8 and R9 may be joined by a single bond to form a cyclopropane; R 10 is hydrogen, and when R8 and R9 do not form a cyclopropane, R8 is a methyl group and R9 is R 10 to form a cyclopropane.

[0009] More preferably, the guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof is the following compound: [ka] [ka] is selected from.

[0010] Preferably, the pharmaceutically acceptable salts of the guaiane-based sesquiterpene derivatives refer to pharmaceutically acceptable salts formed with inorganic or organic acids selected from hydrochlorides, sulfates, phosphates, maleates, fumarates, citrates, etc.

[0011] Furthermore, the pharmaceutically acceptable salts include [ka] is selected from.

[0012] The present application further discloses methods for preparing guaiane-based sesquiterpene derivatives.

[0013] The present application also discloses a pharmaceutical composition comprising, as an active ingredient, a therapeutically effective amount of one or more guaiane-based sesquiterpene derivatives or pharmaceutically acceptable salts thereof, and further comprising a pharmaceutically acceptable carrier, adjuvant, or excipient.

[0014] The present application also discloses the use of the above-mentioned guaiane-based sesquiterpene derivatives or pharmaceutically acceptable salts thereof, and pharmaceutical compositions in the preparation of drugs for preventing or treating NLPR3 inflammasome-associated diseases.

[0015] The present application also discloses the use of the above-mentioned guaiane-based sesquiterpene derivatives or pharmaceutically acceptable salts thereof in combination with other pharmaceutically acceptable therapeutic agents, in particular other NLRP3 inflammasome inhibitors, in the preparation of a medicament for preventing or treating an NLPR3 inflammasome-associated disease.

[0016] The present invention provides a method for preventing or treating an NLPR3 inflammasome-associated disease, which method comprises administering to a patient in need of treatment a therapeutically effective amount of one or more selected from the guaiane-based sesquiterpene derivatives or pharmaceutically acceptable salts thereof according to the present invention, or a pharmaceutical composition containing a therapeutically effective amount of one or more selected from the guaiane-based sesquiterpene derivatives or pharmaceutically acceptable salts thereof according to the present invention as an active ingredient.

[0017] The above-mentioned NLPR3 inflammasome-related diseases include immune diseases, autoimmune diseases, malignant tumors, skin diseases, cardiovascular diseases, liver-related diseases, renal system-related diseases, gastrointestinal tract-related diseases, central nervous system diseases, metabolic diseases, endocrine-related diseases, respiratory diseases, lymphatic system diseases, inflammation, infectious diseases, eye diseases, psychiatric diseases, pain, etc.

[0018] Specifically, (1) cryopyrin protein-associated periodic syndromes (CAPS): Muckle-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS) and chronic infantile neurological cutaneous and articular syndrome (NOMID); (2) autoinflammatory diseases: familial Mediterranean fever (FMF), TNF receptor-associated periodic syndrome (TRAPS), mevalonate kinase deficiency (MKD), hyperimmunoglobulin D and periodic fever syndrome (HIDS), interleukin-1 receptor (DIRA) deficiency, Majeed syndrome, septic arthritis, pyoderma gangrenosum and acne (PAPA), A20 ha (3) Sweet's syndrome: chronic nonbacterial osteomyelitis (CNO), chronic recurrent multifocal osteomyelitis (CRMO), synovitis, acne, impetigo, hyperostosis and osteitis syndrome (SAPHO); (4) autoimmune diseases: multiple sclerosis (MS), type 1 diabetes, psoriasis, rheumatoid arthritis, Behcet's disease, (5) Respiratory diseases: chronic obstructive pulmonary disease (COPD), steroid-resistant asthma, asbestosis, silicosis, and cystic fibrosis; (6) Central nervous system diseases: Parkinson's disease, Alzheimer's disease, motor neuron disease, Huntington's disease, cerebral malaria, and brain damage due to pneumococcal meningitis; (7) Metabolic diseases: type 2 diabetes, atherosclerosis, obesity, gout, and pseudogout; (8) Ophthalmic diseases: ocular epidermis, age-related macular degeneration (AMD), corneal infection, uveitis, and dry eye; (9) Kidney-related diseases: Chronic kidney disease, oxalate nephropathy, and diabetic nephropathy; (10) liver-related diseases: nonalcoholic steatohepatitis and alcoholic liver disease; (11) skin-related inflammatory reactions: contact allergy and sunburn; (12) joint-related inflammatory reactions: osteoarticular, systemic juvenile idiopathic arthritis, adult Still's disease, and relapsing polychondritis; (13) viral infections: dengue virus, Zika virus, influenza, and AIDS virus; (14) hidradenitis suppurativa (HS) and other skin diseases causing cysts; (15) cancers: lung cancer, pancreatic cancer, gastric cancer, myelodysplastic syndrome, abdominal aortic aneurysm, and leukemia;(16) Diseases including polymyositis, colitis, pericarditis, helminth infections, bacterial infections, wound healing, depression, stroke, myocardial infarction, hypertension, Dressler's syndrome, and ischemia-reperfusion injury.

[0019] The colitis includes ulcerative colitis.

[0020] The NLRP3 inflammasome-associated disease includes acute gouty arthritis. [Effects of the Invention]

[0021] The beneficial effects are as follows: Compared with the prior art, this application uses natural ingredients such as parthenolide and dehydrocostus lactone as raw materials, and through tailored structural modifications, synthesizes guaiane sesquiterpene derivatives represented by general formula I with high regioselectivity, and the cyclopropane configurations of the compounds are all α-configuration. Experimental results show that these compounds maintain their inhibitory activity against NLPR3 inflammasome activation, and have significantly improved chemical stability, water solubility, and oral bioavailability, making them more promising for development and application. [Brief explanation of the drawings]

[0022] [Figure 1] This shows the time course of the concentration of Compound 1 and Compound 25 in HEPES 7.4 solution. [Figure 2] 1 shows the time course of Compound 1 and Compound 25 concentrations in mouse plasma. DETAILED DESCRIPTION OF THE INVENTION

[0023] The present application will now be described in detail with reference to specific examples.

[0024] Example 1: Preparation of parthenolide, dehydrocostus lactone (Preparation of Parthenolide) Five kilograms of dried root bark of the sedge orchid was crushed into a coarse powder, soaked in 10 times the amount of 95% ethanol for 12 hours, and extracted twice under reflux for 2 hours each time. The extract was then filtered, the filtrates were combined, concentrated under reduced pressure, and dried to obtain a crude extract of the sedge orchid. This was purified by silica gel column chromatography and run through a petroleum ether-ethyl acetate gradient. The fractions rich in parthenolide and costus lactone were separated, collected, combined, concentrated, and recrystallized to obtain parthenolide, with a preparation yield of 4.0% and a purity of 96.3%. 1 H NMR (500MHz, CDCl3): δ6.31(d,J=2.9Hz,1H),5.62(d,J=3.4Hz,1H),5.20(d,J=11.8Hz,2H),3.85(t,J=8.6Hz,1H),2.78 (d,J=8.9Hz,1H),2.45-2.32(m,2H),2.22-2.10(m,4H),1.70(s,3H),1.69-1.66(m,1H),1.29(s,3H),1.27-1.18(m,1H). ESI-MS(m / z):[M+H] + =249.1(calcd:249.1).

[0025] (Preparation of Dehydrocostus Lactone) 5 kg of southern reed herb was crushed into a coarse powder, soaked in 8 times the amount of petroleum ether for 12 hours, and extracted twice under reflux for 2 hours each time. The extract was filtered, the filtrates were combined, concentrated under reduced pressure, and dried to obtain a southern reed crude extract. The extract was purified by silica gel column chromatography and run through a petroleum ether-ethyl acetate gradient. The dehydrocostus lactone-rich fractions were separated, collected, combined, concentrated, and recrystallized to obtain dehydrocostus lactone in a preparation yield of 1.0% and with a purity of 96.8%. 1H NMR(500MHz): δ6.22(d,J=3.3Hz,1H),5.49(d,J=3.2Hz,1H),5.27(d,J=2.0Hz,1H),5.07(d,J=2.0Hz,1H),4.90(s,1H),4.82(s,1H),3.97-3.94(m, 1H),2.95-2.88(m,2H),2.87(dd,J=9.3,3.0Hz,1H),2.24-2.22(m,1H),2. 16-2.13(m,1H),1.99-1.96(m,2H),1.88-1.86(m,2H),1.42-1.40(m,2H). ESI-MS(m / z):[M+H] + =231.1(calcd:231.1).

[0026] Example 2: Synthesis of Compounds 1 to 9 Although arglabin has a strong inhibitory effect on NLPR3 inflammasome activation, the epoxy ring in its structure can undergo hydrolysis under acidic conditions, and tests have shown that the degradation rate reaches 50% within 8 hours. Therefore, we hope to replace the epoxy ring in arglabin with a cyclopropane to improve its chemical stability while maintaining its activity.

[0027] (Synthesis of Compound 1) [ka]

[0028] Dichloromethane (50 mL), p-toluenesulfonic acid (125 mg, 0.73 mmol), and parthenolide (5 g, 20.16 mmol) were added sequentially to a 150 mL round-bottom flask and stirred at room temperature. The reaction was monitored for completion by TLC. The reaction mixture was washed with water (10 mL x 3) and saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography to obtain the intermediate Micheliolide (MCL) in 90% yield. 1H NMR (500MHz, CDCl3): δ6.21(d,J=3.5Hz,1H),5.51(d,J=3.0Hz,1H),3.81(t,J=10.5Hz,1H),2.73(d,J=10.5Hz,1H),2.68-2.64( m,2H),2.42-2.37(m,1H),2.26-2.16(m,3H),2.11-2.08(m,1H),1.83-1.75(m,2H),1.69(s,3H),1.31(s,3H),1.27-1.25(m,1H). ESI-MS(m / z):[M+Na] + =271.1(calcd:271.1).

[0029] Ethylene glycol dimethyl ether (1.67 mL, 21.26 mmol) was added to anhydrous dichloromethane (67 mL) in an ice bath under nitrogen protection. After stirring uniformly, 13.3 mL of diethylzinc solution (1 M n-hexane solution) was added, and methylene iodide (2.67 mL, 3.11 mmol) was slowly added dropwise. The mixture was stirred for 10 min to prepare the cyclopropanation reagent. Another round-bottom flask was added to anhydrous dichloromethane (5 mL) with Micheliolide (MCL) (300 mg, 1.21 mmol). The mixture was stirred until dissolved, then protected with nitrogen gas and placed in an ice bath. The cyclopropanation reagent was added dropwise to the substrate solution, and the reaction was continued for 1 hour after the addition was complete. The mixture was then transferred to room temperature and allowed to react overnight. The reaction mixture was quenched with saturated ammonium chloride, filtered, washed with water (10 mL × 3) and saturated brine (10 mL × 3) in that order, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography to obtain compound 1 in a 75% yield. 1 H NMR (500MHz, CDCl3): δ6.14(d,J=3.5Hz,1H),5.45(d,J=3.0Hz,1H),3.82(t,J=10.5Hz,1H),2.52-2.48(m,1H),2.26-2.21(m,1H),2.09-1.99(m,2H) ,1.93-1.89(m,1H),1.88-1.86(m,1H,H5),1.85-1.83(m,1H),1.55(s,3H) ,1.52-1.44(m,2H),1.26-1.14(m,2H),1.11(s,3H),0.81(d,J=4.0Hz,1H,H16a ), 0.54(d, J = 4.0 Hz, 1H, H 16b ). The ROESY spectrum shows the H5 and H 16a The signal correlation was confirmed, confirming that the cyclopropane was in the α-configuration. ESI-MS (m / z): [M+Na] + =285.2(calcd:285.2).

[0030] (Preparation of Compound 2) [ka]

[0031] Compound 1 (200 mg, 0.81 mmol) and anhydrous pyridine (10 mL) were sequentially added to a round-bottom flask, protected with nitrogen gas, and dissolved in an ice-water bath. Phosphoric acid trichloride (1242 mg, 8.10 mmol) was added dropwise, and the mixture was allowed to react at room temperature for 2 h. The reaction mixture was poured into ice-water and extracted with ethyl acetate (10 mL x 3). The organic layer was washed sequentially with saturated copper sulfate solution (10 mL x 6), water (10 mL x 3), and saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography to obtain compound 2 in 65% yield. 1 H NMR (500MHz, CDCl3): δ6.14(d,J=3.5Hz,1H),5.59-5.57(m,1H),5.44(d,J=3.0Hz,1H),3.90(t,J=10.5Hz,1H),2.74-2.71(m,1H),2.51-2.46(m,1H) ,2.32-2.28(m,2H),2.04-2.00(m,1H),1.97(s,3H),1.81-1.77(m,1H,H5) ,1.54-1.46(m,1H),1.18-1.14(m,1H),1.13(s,3H),0.61(d,J=4.0Hz,1H,H 16a ), 0.50(d, J = 4.0 Hz, 1H, H 16b ). The ROESY spectrum is 16a The signal correlation was confirmed, confirming that the cyclopropane was in the α-configuration. ESI-MS (m / z): [M+Na] + =267.1(calcd:267.2).

[0032] (Preparation of Compound 3) [ka]

[0033] Metachloroperbenzoic acid (267.5 mg, 1.55 mmol) and anhydrous dichloromethane (20 mL) were added to a round-bottom flask, followed by the gradual addition of a dichloromethane solution (5 mL) of compound 2 (248 mg, 1.00 mmol). The mixture was stirred overnight. After quenching with saturated sodium thiosulfate solution, the mixture was washed sequentially with saturated sodium bicarbonate (10 mL x 3), water (10 mL x 3), and saturated brine (10 mL x 3). After drying over anhydrous sodium sulfate, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography to give compound 3-1 in 83% yield. 1 H NMR(500MHz,CDCl3):δ6.18(d,J=3.5Hz,1H),5.48(d,J=3.0Hz,1H),3.79(t,J=10.5Hz,1H,H6),2.53-2.48(m,1H),2.28-2.24(m,1H, H3),2.16-2.10(m,2H),2.04-2.01(m,1H,H5),1.73(s,3H),1.56-1.45(m,3H),1.12-1.09(m,1H),1.08(s,3H),0.57(d,J=4.0Hz,1H,H 16a ), 0.49(d,J=4.0Hz,1H,H 16b ). The ROESY spectrum shows the H5 and H 16a , H3 and H6 showed a signal correlation, confirming that cyclopropane was in the α-configuration and propylene oxide was in the α-configuration. ESI-MS (m / z): [M+Na] + =283.1(calcd:283.1).

[0034] The intermediate 3-1 (260 mg, 1.00 mmol), methanol (10 mL), and paratoluenesulfonic acid (172 mg, 1.00 mmol) were sequentially added to a round-bottom flask and stirred overnight. The reaction mixture was concentrated and extracted with ethyl acetate (10 mL x 3). The organic layer was washed with saturated sodium bicarbonate (10 mL x 3), water (10 mL x 3), and saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain compound 3 in 85% yield. 1 H NMR(500MHz,CDCl3):δ6.15(d,J=3.5Hz,1H),5.44(d,J=3.0Hz,1H),4.25(t,J=10.5Hz,1H,H6),3.64-3.62(m,1H,H3),3.41(s,3H,H 17 ),2.55-2.48(m,1H,H7),2.41(s,1H),2.22-2.18(m,1H),2.08(d,J=11.0Hz,1H),2.06-2.01(m,1H),1.98-1.94(m, 1H),1.80-1.75(m,1H,H5),1.61-1.55(m,1H),1.49(s,3H),1.47-1.42(m,1H),1.10(s,3H),0.57(d,J=4.0Hz,1H,H 16a ), 0.49(d,J=4.0Hz,1H,H 16b ). ROESY is H3 and H6, H5 and H 16a , H7 and H 17 The signal correlation was confirmed, confirming that the cyclopropane was in the α-configuration and that the 3-OH and 4-OMe were in the α-configuration. ESI-MS (m / z): [M+Na] + =315.1(calcd:315.1).

[0035] (Preparation of Compound 4) [ka]

[0036] A round-bottom flask was charged with tetrahydrofuran (5 mL) and water (1 mL), followed by the addition of paratoluenesulfonic acid (272 mg, 1.00 mmol) and intermediate 3-1 (260 mg, 1.00 mmol) and stirring overnight. The reaction mixture was concentrated and extracted with ethyl acetate (10 mL x 3). The organic layer was washed with saturated sodium bicarbonate (10 mL x 3), water (10 mL x 3), and saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to give compound 4 in 47% yield. 1 H NMR(500MHz,CDCl3):δ6.16(d,J=3.5Hz,1H),5.46(d,J=3.0Hz,1H),4.33(t,J=10.5Hz,1H,H6),4.12-4.09(m,1H,H3),2.54-2.47(m,1H,H6),2.24 -2.19(m,1H),2.10-2.02(m,4H),2.00(d,J=5.5Hz,1H),1.87-1.83(m,1H ,H5),1.61-1.54(m,2H),1.51(s,3H),1.12(s,3H),0.80(d,J=4.0Hz,1H,H 16a ), 0.50(d, J = 4.0 Hz, 1H, H 16b ). The ROESY spectrum shows the difference between H3 and H6, H6 and H 14 , and H5 and H 16a The signal correlation was confirmed, confirming that the cyclopropane was in the α-configuration and that the 3-OH and 4-OH were in the α-configuration. ESI-MS (m / z): [M+Na] + =301.2(calcd:301.2).

[0037] (Preparation of Compound 5) [ka]

[0038] Compound 1 (124 mg, 0.50 mmol) and dichloromethane (5 mL) were placed in a round-bottom flask, protected with nitrogen gas, and placed at -78 °C. The flask was stirred and dissolved, and DAST reagent (161 mg, 1.00 mmol) was slowly added dropwise. Stirring was continued for 10-15 minutes after the addition. The reaction mixture was quenched with water, diluted with dichloromethane, washed with water (10 mL x 3) and saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to give compound 5 in 55% yield. 1 H NMR (500MHz, CDCl3): δ6.14(d,J=3.5Hz,1H),5.43(d,J=3.0Hz,1H),4.11(t,J=10 .5Hz,1H),2.45-2.40(m,1H),2.35-2.31(m,1H),2.23-2.17(m,1H),2.09-2.03(m, 2H),2.02-1.97(m,1H),1.85-1.74(m,1H,H5),1.65(d,J=21.5Hz,3H),1.54-1.46 (m,1H),1.24-1.20(m,1H),1.17(s,3H),1.15-1.12(m,1H),0.80(d,J=4.0Hz,1H,H 16a ), 0.50(d, J = 4.0 Hz, 1H, H 16b ). 19 FNMR (470 MHz, CDCl3): δ-148.17. ROESY spectrum shows H5 and H 16a The signal correlation was confirmed, confirming that the cyclopropane was in the α-configuration. ESI-MS (m / z): [M+Na] + =287.2(calcd:287.2).

[0039] (Preparation of Compound 6) [ka]

[0040] A round-bottom flask was charged with Burgess Reagent (282 mg, 1.10 mmol) and tetrahydrofuran anhydride (10 mL), protected with nitrogen gas, and treated with an ice bath. Compound 1 (262 mg, 1.00 mmol) was added and stirred for 20 minutes, then the temperature was returned to room temperature and stirring was continued for 3 hours. The reaction mixture was concentrated and extracted with ethyl acetate (10 mL x 3). The organic layer was washed successively with water (10 mL x 3) and saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain compound 6 in 75% yield. 1 H NMR(500MHz, CDCl3): δ6.14(d,J=3.5Hz,1H),5.60-5.58(m,1H),5.44(d,J=3.0 Hz,1H),3.90(t,J=10.0Hz,1H),2.75-2.71(m,1H),2.52-2.46(m,1H),2.32-2. 29(m,2H),2.05-2.00(m,1H),1.98(s,3H),1.81-1.77(m,1H),1.55-1.47(m,1H )1.18-1.14(m,1H),1.13(s,3H),0.63(d,J=5.0Hz,1H),0.51(d,J=5.0Hz,1H). ESI-MS(m / z):[M+Na] + =267.3(calcd:267.3).

[0041] (Preparation of Compound 7) [ka]

[0042] Compound 1 (262 mg, 1.00 mmol) was added to a round-bottom flask, protected with nitrogen gas, and 3 mL of anhydrous dichloromethane and triethylamine (2.7 g, 27.0 mmol) were added sequentially. The mixture was then cooled to room temperature and allowed to react overnight. The reaction mixture was poured into ice water and extracted with ethyl acetate (10 mL x 3). The organic layer was washed with water (10 mL x 3) and saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to give compound 7 in 51% yield. 1H NMR(500MHz, CDCl3): δ6.16(d,J=3.5Hz,1H),5.47(d,J=3.0Hz,1H),3.82(t,J=1 0.5Hz,1H),2.52-2.48(m,1H),2.27-2.23(m,2H),2.21-2.02(m,2H),1.95-1.92 (m,1H),1.90-1.88(m,1H),1.87-1.85(m,1H,H5),1.57(s,3H),1.53-1.49(m,2H ),1.24(t,J=10.0Hz,3H),1.23-1.14(m,2H),1.11(s,3H),0.59(d,J=4.0Hz,1H,H 16a ), 0.38(d, J = 4.0 Hz, 1H, H 16b ). ROESY is H5 and H 16a The signal correlation was confirmed, confirming that the cyclopropane was in the α-configuration. ESI-MS (m / z): [M+Na] + =341.2(calcd:341.2).

[0043] (Preparation of Compound 8) [ka]

[0044] Ethylene glycol dimethyl ether (1.67 mL, 21.26 mmol) was added to anhydrous dichloromethane (67 mL) in an ice bath under nitrogen protection. After uniform stirring, 13.3 mL of diethylzinc solution (1 M n-hexane solution) was added, and methylene iodide (2.67 mL, 3.11 mmol) was slowly added dropwise. The mixture was stirred for 10 minutes to prepare the cyclopropanation reagent. In a separate round-bottom flask, dehydrocostus lactone (300 mg, 1.21 mmol) and anhydrous dichloromethane (5 mL) were added and dissolved by stirring. The mixture was then protected with nitrogen gas and placed in an ice bath. The cyclopropanation reagent was added dropwise to the substrate solution. After the addition was complete, the reaction was continued for 1 hour, then transferred to room temperature and allowed to react overnight. The reaction mixture was quenched with saturated ammonium chloride, filtered, washed with water (10 mL × 3) and saturated brine (10 mL × 3) in that order, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by silica gel column chromatography to obtain compound 8 in a 65% yield. 1 H NMR(500MHz,CDCl3):δ6.24(d,J=3.5Hz,1H),5.45(d,J=3.0Hz,1H),4.24(dd,J=10.8, 8,8Hz,1H),2.79-2.74(m,1H),2.22-2.20(m,1H),2.10-2.06(m,1H),1.95(dd,J=10.4 ,8,8Hz,1H),1.73-1.67(m,2H),1.63-1.57(m,2H),1.50-1.45(m,1H),1.38-1.35(m,2 H),0.98(s,1H),0.64(s,1H),0.50-0.48(m,1H),0.42-0.40(m,1H),0.37-0.27(m,4H). ESI-MS(m / z):[M+Na] + =259.4(calcd:259.4).

[0045] Preparation of Compound 9 [ka]

[0046] Under nitrogen protection in an ice bath, 5 mL of diethylzinc solution (1 M n-hexane solution) was added to 2.5 mL of dichloromethane. Trifluoroacetic acid (570 mg, 5.00 mmol) was dissolved in 0.8 mL of dichloromethane and added dropwise to the solution. Stirred for 20 min. Methylene iodide (1.34 g, 5.00 mmol) was dissolved in 0.8 mL of dichloromethane and added dropwise to the solution. Stirred for 20 min. MCL (248 mg, 1.00 mmol) was dissolved in 0.8 mL of dichloromethane and added to the solution. Stirred for 5 h. The reaction mixture was quenched with saturated ammonium chloride and extracted with ethyl acetate (10 mL x 3). The organic layer was washed sequentially with water (10 mL x 3) and saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain compound 9 in 12% yield. 1 H NMR (500MHz, CDCl3): δ6.14(d,J=3.5Hz,1H),5.44(d,J=3.0Hz,1H),3.83(t,J=10.5Hz,1H),3.23(s,3H),2.50-2.46(m,1H),2.28-2.23( m,1H),2.02-1.97(m,4H),1.78-1.71(m,1H,H5),1.53(s,3H),1.51-1.39(m,2H),1.13-1.07(m,1H),1.09(s,3H),0.70(d,J=4.0Hz,1H,H 16a ), 0.46(d, J = 4.0 Hz, 1H, H 16b ). The ROESY spectrum shows the H5 and H 16a The signal correlation was confirmed, confirming that the cyclopropane was in the α-configuration. ESI-MS (m / z): [M+Na] + =299.2(calcd:299.2).

[0047] Example 3: Stability test of compounds 1 to 9 2.00 mg of compounds 1-9 were precisely weighed and dissolved in 500 μL of chromatographic methanol. 1500 μL of artificial gastric juice was added, mixed uniformly, and then sonicated for complete dissolution. 50 μL of the solution was precisely aspirated, diluted with 200 μL of chromatographic methanol, filtered, and analyzed by HPLC (HPLC analysis conditions: mobile phase: 65% methanol-35% water, flow rate: 0.8 mL / min, column temperature: 25°C). The initial peak area was recorded. The solution was placed in a constant-temperature water bath at 37°C, and samples were taken at 8, 16, and 24 hours, respectively, for HPLC analysis. The peak areas were calculated to obtain stability data for the sample under the artificial gastric juice environment for 8, 16, and 24 hours.

[0048] As shown in Table 1, the stability of compounds 1 to 9 was significantly improved compared to arglabin and dehydrocostus lactone.

[0049] [Table 1]

[0050] Example 4: Prodrug Preparation (Prodrugs Contain Salts) (Preparation of Compound 10) [ka]

[0051] Compound 1 (262 mg, 1.00 mmol), dichloromethane (30 mL), dimethylamine hydrochloride (815 mg, 10.00 mmol), and potassium carbonate (2764 mg, 20 mmol) were added to a round-bottom flask and stirred for 4 hours. The reaction mixture was filtered, washed with water (10 mL x 3) and saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether:ethyl acetate:triethylamine = 1:1:0.02) to obtain compound 10 in 85% yield. 1H NMR (500MHz, CDCl3): δ3.92(t,J=10.5Hz,1H),3.55-3.47(m,1H),3.33-3.29(m,1H), 2.38-2.34(m,1H),2.28(s,6H),2.21-2.16(m,2H),1.98(d,J=15.0Hz,2H),1.88(t,J =5.0Hz,2H),1.72(d,J=10.0Hz,1H),1.60-1.52(m,2H),1.50(s,3H),1.20(t,J=5.0H z,1H),1.12(s,3H),1.08-1.04(m,1H),0.76(d,J=5.0Hz,1H),0.53(d,J=5.0Hz,1H). ESI-MS(m / z):[M+Na] + =330.2(calcd:330.2).

[0052] (Preparation of Compound 11) [ka]

[0053] Compound 2 and dimethylamine hydrochloride were used as starting materials, and the preparation method was the same as for Compound 10. The yield of Compound 11 was 80%. 1 H NMR(500MHz,CDCl3):δ5.52-5.46(m,1H),3.98(t,J=10.0Hz,1H),3.41-3.32(m,1H),3 .20-3.16(m,1H),2.98-2.92(m,1H),2.28(s,6H),2.22-2.19(m,1H),2.10(d,J=10.0Hz ,1H),2.06-2.02(m,1H),1.98-1.94(m,1H),1.91(s,3H),1.84-1.80(m,2H),1.62-1.5 6(m,1H),1.19(s,3H),1.14-1.08(m,1H),0.69(d,J=5.0Hz,1H),0.51(d,J=5.0Hz,1H). ESI-MS(m / z):[M+Na] + =312.2(calcd:312.2).

[0054] (Preparation of Compound 12) [ka]

[0055] Compound 3 and dimethylamine hydrochloride were used as starting materials, and the preparation method was the same as for Compound 10. The yield of Compound 12 was 95%. 1 H NMR(500MHz, CDCl3): δ3.96(t,J=10.0Hz,1H),3.28-3.23(m,1H),3.15-3.10(m ,1H),2.94(s,3H),2.70-2.65(m,1H),2.28(s,6H),2.03-1.99(m,1H),1.91-1. 88(m,1H),1.86-1.76(m,4H),1.62-1.56(m,1H),1.48(s,3H),1.28-1.24(m,1H ),1.12(s,3H),1.09-1.05(m,1H),0.70(d,J=5.0Hz,1H),0.54(d,J=5.0Hz,1H). ESI-MS(m / z):[M+Na] + =360.4(calcd:360.5).

[0056] (Preparation of Compound 13) [ka]

[0057] Compound 4 and dimethylamine hydrochloride were used as starting materials, and the preparation method was the same as for Compound 10. The yield of Compound 13 was 90%. 1 H NMR (500MHz, CDCl3): δ3.94(t,J=10.0Hz,1H),3.38-3.36(m,1H),3.23-3.18(m,1H), 3.19-3.15(m,2H),2.78-2.70(m,1H),2.28(s,6H),2.13-2.09(m,1H),1.97-1.92(m,1 H),1.90-1.86(m,1H),1.82-1.79(m,1H),1.77-1.75(m,1H),1.62-1.59(m,1H),1.51 (s,3H),1.18(s,3H),1.11-1.07(m,1H),0.70(d,J=5.0Hz,1H),0.48(d,J=5.0Hz,1H). ESI-MS(m / z):[M+Na] +=368.4(calcd:368.4).

[0058] (Preparation of Compound 14) [ka]

[0059] Compound 5 and dimethylamine hydrochloride were used as starting materials, and the preparation method was the same as for Compound 10. The yield of Compound 14 was 83%. 1 H NMR(500MHz, CDCl3): δ3.96(t,J=10.0Hz,1H),3.35-3.30(m,2H),2.80-2.76(m ,1H),2.30(s,6H),2.29-2.19(m,2H),2.08(t,J=10.0Hz,1H),1.96-1.90(m,1H) ,1.84-1.82(m,2H),1.81-1.77(m,2H),1.58(d,J=21.5Hz,3H)1.46-1.39(m,1H ),1.18(s,3H),1.16-1.12(m,1H),0.62(d,J=5.0Hz,1H),0.58(d,J=5.0Hz,1H). ESI-MS(m / z):[M+Na] + =332.4(calcd:332.4).

[0060] (Preparation of Compound 15) [ka]

[0061] Compound 6 and dimethylamine hydrochloride were used as starting materials, and the preparation method was the same as for Compound 10. The yield of Compound 15 was 91%. 1H NMR (500MHz, CDCl3): δ3.99(t,J=10.0Hz,1H),3.41-3.36(m,1H),3.20-3.18(m, 1H),2.82-2.77(m,1H),2.28(s,6H),2.22-2.19(m,1H),2.16-2.10(m,2H),1.98- 1.94(m,1H),1.91(s,3H),1.84-1.80(m,1H),1.78-1.75(m,1H),1.62-1.56(m,2 H),1.15(s,3H),1.11-1.07(m,1H),0.64(d,J=5.0Hz,1H),0.50(d,J=5.0Hz,1H). ESI-MS(m / z):[M+Na] + =312.4(calcd:312.4).

[0062] (Preparation of Compound 16) [ka]

[0063] Compound 7 and dimethylamine hydrochloride were used as starting materials, and the preparation method was the same as for Compound 10. The yield of Compound 16 was 80%. 1 H NMR (500MHz, CDCl3): δ3.92(t,J=10.0Hz,1H),3.40-3.30(m,2H),2.83-2.79(m,1H),2.30( s,6H),2.19-2.13(m,2H),2.04-1.97(m,2H),1.95-1.90(m,2H),1.86-1.82(m,1H),1.76-1. 73(m,1H),1.60(s,3H),1.56-1.52(m,1H),1.33-1.25(m,1H),1.14(s,3H),1.02(t,J=10.0 Hz,3H),1.00-0.95(m,1H),0.88-0.82(m,1H),0.48(d,J=5.0Hz,1H),0.40(d,J=5.0Hz,1H). ESI-MS(m / z):[M+Na] + =386.5(calcd:386.5).

[0064] (Preparation of Compound 17) [ka]

[0065] Compound 8 and dimethylamine hydrochloride were used as starting materials, and the preparation method was the same as for Compound 10. The yield of Compound 17 was 78%. 1 H NMR(500MHz,CDCl3):δ4.15(dd,J=9.6,10.0Hz,1H),2.70(dd,J=12.8,4.8Hz,1H),2 .48(dd,J=12.8,4.8Hz,1H),2.36-2.28(m,2H),2.22(s,6H),2.02-1.98(m,2H),1.8 4-1.80(m,1H),1.70-1.67(m,1H),1.45-1.30(m,5H),1.11-1.06(m,1H),1.01-0.98 (m,1H),0.70-0.68(m,1H),0.46-0.42(m,1H),0.37-0.28(m,4H),0.15-0.11(m,2H). ESI-MS(m / z):[M+Na] + =327.3(calcd:327.3).

[0066] (Preparation of Compound 18) [ka]

[0067] Compound 1 and piperidine were used as starting materials, and the preparation method was the same as for Compound 10. The yield of Compound 18 was 89%. 1 H NMR (500MHz, CDCl3): δ3.79(t,J=10.5Hz,1H),2.79-2.75(m,1H),2.56-2 .53(m,1H),2.46-2.40(m,3H),2.39-2.32(m,3H),2.19-2.08(m,4H),1.94 -1.85(m,3H),1.77(d,J=10.5Hz,1H),1.54(s,6H),1.45-1.39(m,4H),1.1 2(s,3H),1.09-1.04(m,1H),0.79(d,J=4.0Hz,1H),0.52(d,J=4.0Hz,1H). ESI-MS(m / z):[M+Na] + =326.3(calcd:326.3).

[0068] (Preparation of Compound 19) [ka]

[0069] Compound 1 and Pyrrolidine was used as the starting material, and the preparation method was the same as for Compound 10. The yield of Compound 19 was 87%. 1 H NMR (500MHz, CDCl3): δ3.79(t,J=10.5Hz,1H),2.89-2.80(m,2H),2.57-2.51(m,4H),2.37-2.32(m,1H),2.15-2.06(m,3H),1.94-1.87( m,3H),1.78-1.76(m,5H),1.55(s,3H),1.51-1.39(m,3H),1.11(s,3H),1.09-1.03(m,1H),0.77(d,J=4.0Hz,1H),0.51(d,J=4.0Hz,1H). ESI-MS(m / z):[M+Na] + =356.3(calcd:356.2).

[0070] (Preparation of Compound 20) [ka]

[0071] Compound 1 and Morpholine was used as the starting material, and the preparation method was the same as for Compound 10. The yield of Compound 20 was 76%. 1H NMR (500MHz, CDCl3): δ3.81(t,J=10.5Hz,1H),3.74-3.67(m,3H),2.82-2.79(m,1H),2. 65-2.61(m,1H),2.53-2.44(m,5H),2.40-2.35(m,1H),2.21-2.17(m,1H),2.13-2.07(m, 2H),1.95-1.86(m,3H),1.78(d,J=10.5Hz,1H),1.55(s,3H),1.52-1.39(m,2H),1.31-1. 27(m,1H),1.12(s,3H),1.09-1.04(m,1H),0.80(d,J=4.0Hz,1H),0.54(d,J=4.0Hz,1H). ESI-MS(m / z):[M+Na] + =372.2(calcd:372.2).

[0072] (Preparation of Compound 21) [ka]

[0073] Compound 1 and piperazine were used as starting materials, and the preparation method was the same as for Compound 10. The yield of Compound 21 was 74%. 1 H NMR (500MHz, CDCl3): δ3.79(t,J=10.5Hz,1H),2.82-2.79(m,1H),2.64-2.60(m,1H),2 .55-2.42(m,5H),2.38-2.34(m,2H),2.29(s,3H),2.20-2.16(m,1H),2.15-2.05(m,2H) ,1.93-1.83(m,3H),1.76(d,J=10.5Hz,1H),1.54(s,3H),1.49-1.38(m,2H),1.29-1.2 5(m,1H),1.12(s,3H),1.09-1.03(m,1H),0.78(d,J=4.0Hz,1H),0.52(d,J=4.0Hz,1H). ESI-MS(m / z):[M+Na] + =371.2(calcd:371.2).

[0074] (Preparation of Compound 22) [ka]

[0075] Compound 1 and piperazine were used as starting materials, and the preparation method was the same as for Compound 10. The yield of Compound 22 was 74%. 1 H NMR (500MHz, CDCl3): δ3.79(t,J=10.5Hz,1H),2.83-2.74(m,3H),2.59-2.55(m ,1H),2.39-2.35(m,2H),2.20-2.07(m,4H),1.96-1.83(m,4H),1.78(d,J=10.5H z,1H),1.55(s,3H),1.51-1.23(m,5H),1.21-1.15(m,2H),1.13(s,3H),1.10-1 .05(m,1H),0.93(d,J=6.5Hz,3H),0.79(d,J=4.0Hz,1H),0.52(d,J=4.0Hz,1H). ESI-MS(m / z):[M+Na] + =384.3(calcd:384.3).

[0076] (Preparation of Compound 23) [ka]

[0077] Compound 23 was prepared from compound 1 and N-Boc-piperazine as starting materials in the same manner as compound 10, with a yield of 65%. 1H NMR (500MHz, CDCl3): δ3.81(t,J=10.5Hz,1H),2.83-2.79(m,1H),2.66-2.62(m,2H),2.42- 2.35(m,4H),2.21-2.16(m,1H),2.13-2.07(m,2H),1.94-1.84(m,3H),1.78(d,J=10.5Hz,1H ),1.55(s,3H),1.48(s,9H),1.46-1.44(m,1H),1.40-1.39(m,1H),1.32-1.25(m,3H),1.13 (s,3H),1.10-1.04(m,2H),0.92-0.85(m,1H),0.80(d,J=4.0Hz,1H),0.54(d,J=4.0Hz,1H). ESI-MS(m / z):[M+Na] + =471.3(calcd:471.3).

[0078] (Preparation of Hydrochloride Salt 24 of Compound 10) [ka]

[0079] Compound 10 (307 mg, 1 mmol) was dissolved in dichloromethane (2 mL), stirred at room temperature for 2 hours, and then hydrochloric acid solution was added dropwise to adjust the pH to 4-5. The solution was filtered, and the resulting solid was washed with dichloromethane. The resulting white solid was the hydrochloride salt of compound 10 (compound 24) in a 90% yield. 1 H NMR(500MHz,CD3OD):δ4.15-4.10(m,1H,H6),3.42-3.37(m,1H),3.31-3.26(m,1H),3.04-2.98(m,1H,H 11 ),2.91(s,6H),2.21-2.12(m,2H),2.02(d,J=15.0Hz,2H),1.88(t,J=5.0Hz,2H),1.78(d,J=10.0Hz,1H,H5), 1.64-1.54(m,2H),1.52(s,3H),1.26(t,J=5.0Hz,1H),1.14(s,3H),1.10-1.05(m,1H),0.76(d,J=5.0Hz,1H,H 16a ), 0.53(d, J = 5.0 Hz, 1H, H 16b). The ROESY spectrum shows the H5 and H 16a hydrogen, and H6 and H 11 The hydrogens in the cyclopropane and 11-H were shown to have signal correlation, confirming that the cyclopropane was in the α-configuration and the 11-H was in the β-configuration. ESI-MS (m / z): [M+H] + =344.9(calcd:344.9).

[0080] (Preparation of fumarate salt 25 of compound 10) [ka]

[0081] Using fumarate instead of hydrochloric acid, fumarate compound 25 was prepared by following the preparation method of the hydrochloride salt of compound 10. The yield was 80%. 1 H NMR(500MHz,CD3OD):δ6.72(s,2H),4.13(t,J=10.0Hz,1H,H6),3.41-3.39(m,1H),3.30-3.27(m,1H),3.03-2.98(m,1H,H 11 ),2.91(s,6H),2.21-2.12(m,2H),1.94-1.91(m,1H),1.88(t,J=5.0Hz,2H),1.84-1.80(m,1H),1.78(d,J =15.0Hz,1H,H5),1.64-1.54(m,2H),1.52(s,3H),1.14(s,3H),1.10-1.04(m,1H),0.75(d,J=5.0Hz,1H,H 16a ), 0.52(d, J = 5.0 Hz, 1H, H 16b ). The ROESY spectrum shows the H5 and H 16a , and H6 and H 11 The signal correlation was confirmed, confirming that the cyclopropane was in the α-configuration and the 11-H was in the β-configuration. ESI-MS (m / z): [M+H] + =424.5(calcd:424.5).

[0082] (Preparation of Hydrochloride Salt 26 of Compound 11) [ka]

[0083] Using compound 11 as a starting material, compound 26 could be prepared in 95% yield by following the preparation method of the hydrochloride salt of compound 10. 1 H NMR(500MHz,CD3OD):δ5.62-5.58(m,1H),4.19(t,J=10.0Hz,1H,H6),3.51-3.46(m,1H),3.40-3.36(m,1H),3.12-3.07(m,1H,H 11 ),2.98(s,6H),2.82-2.79(m,1H),2.30(d,J=10.0Hz,1H),2.26-2.12(m,1H),1.98-1.94(m,1H),1.91(s,3H),1. 84-1.82(m,2H),1.80-1.78(m,1H,H5),1.66-1.59(m,1H),1.17(s,3H),1.16-1.10(m,1H),0.61(d,J=5.0Hz,1H,H 16a ), 0.48(d, J = 5.0 Hz, 1H, H 16b ). The ROESY spectrum shows the H5 and H 16a , and H6 and H 11 The signal correlation was confirmed, confirming that the cyclopropane was in the α-configuration and the 11-H was in the β-configuration. ESI-MS (m / z): [M+H] + =326.9(calcd:326.9).

[0084] (Preparation of Hydrochloride Salt 27 of Compound 12) [ka]

[0085] Using compound 12 as a starting material, compound 27 could be prepared in 95% yield by following the preparation method of the hydrochloride salt of compound 10. 1 H NMR(500MHz,CD3OD):δ4.39(t,J=10.0Hz,1H,H6),3.48-3.43(m,1H,H7),3.40-3.36(m,1H),3.34(s,3H,H 17 ), 3.10-3.05(m,1H,H 11),2.98(s,6H),2.23-2.19(m,1H,H3),2.11-2.08(m,1H),1.87-1.80(m,3H),1.79-1.77(m,1H,H5),1.6 5-1.56(m,1H),1.51(s,3H),1.27-1.25(m,1H),1.15(s,3H),1.13-1.08(m,1H),0.74(d,J=5.0Hz,1H,H 16a ), 0.51(d, J = 5.0 Hz, 1H, H 16b ). ESI-MS(m / z):[M+H] + =374.9(calcd:374.9).

[0086] (Preparation of Hydrochloride Salt 28 of Compound 13) [ka]

[0087] Using compound 13 as a starting material, compound 28 could be prepared in 85% yield by following the preparation method of the hydrochloride salt of compound 10. 1 H NMR(500MHz,CD3OD):δ4.47(t,J=10.0Hz,1H,H6),3.88-3.86(m,1H),3.63-3.58(m,1H),3.49-3.45(m,2H),3.10-3.06(m,1H,H 11 ),3.00(s,6H),2.23-2.19(m,1H),1.97-1.92(m,1H),1.90-1.81(m,3H),1.66-1.59(m,1H,H5 ),1.52(s,3H),1.14(s,3H),1.12-1.10(m,1H),0.73(d,J=5.0Hz,1H),0.50(d,J=5.0Hz,1H). ESI-MS(m / z):[M+H] + =360.9(calcd:360.9).

[0088] (Preparation of Hydrochloride Salt 29 of Compound 14) [ka]

[0089] Using compound 14 as a starting material, compound 29 could be prepared in 83% yield by following the preparation method of the hydrochloride salt of compound 10. 1 H NMR(500MHz,CD3OD):δ4.36(t,J=10.0Hz,1H,H6),3.50-3.40(m,2H),3.15-3.11(m,1H,H 11 ),3.00(s,6H),2.29-2.19(m,2H),2.11(t,J=10.0Hz,1H),2.06-2.00(m,1H),1.94-1.87(m,2H),1.84-1.82(m,1H),1 .81-1.78(m,1H,H5)1.60(d,J=21.5Hz,3H)1.60-1.51(m,1H),1.20(s,3H),1.18-1.15(m,1H),0.54(d,J=5.0Hz,1H,H 16a ), 0.50(d, J = 5.0 Hz, 1H, H 16b ). ESI-MS(m / z):[M+H] + =346.9(calcd:346.9).

[0090] (Preparation of Hydrochloride Salt 30 of Compound 15) [ka]

[0091] Using compound 15 as a starting material, compound 30 could be prepared in 80% yield by following the preparation method of the hydrochloride salt of compound 10. 1 H NMR(500MHz,CD3OD):δ4.19(t,J=10.0Hz,1H,H6),3.51-3.46(m,1H),3.40-3.36(m,1H),3.12-3.07(m,1H,H 11 ),2.98(s,6H),2.82-2.79(m,1H),2.26-2.12(m,2H),1.98-1.94(m,1H),1.91(s,3H),1.84-1.80(m, 2H),1.66-1.59(m,2H),1.17(s,3H),1.16-1.10(m,1H),0.61(d,J=5.0Hz,1H),0.48(d,J=5.0Hz,1H). ESI-MS(m / z):[M+H] +=326.9(calcd:326.9).

[0092] (Preparation of Hydrochloride Salt 31 of Compound 16) [ka]

[0093] Using compound 16 as a starting material, compound 31 could be prepared in 76% yield by following the preparation method of the hydrochloride salt of compound 10. 1 H NMR(500MHz,CD3OD):δ4.22(t,J=10.0Hz,1H,H6),3.50-3.38(m,2H),3.13-3.09(m,1H,H 11 ),2.99(s,6H),2.69-2.63(m,2H),2.24-2.17(m,2H),1.99-1.93(m,2H),1.86-1.82(m,1H,H5),1.68-1.59(m,1H),1.54-1.45(m,1H) ),1.36-1.30(m,1H),1.27(s,3H),1.14(s,3H),1.06(t,J=10.0Hz,3H),1.04-1.00(m,1H),0.93-0.88(m,1H),0.42(d,J=5.0Hz,1H,H 16a ), 0.36(d, J = 5.0 Hz, 1H, H 16b ). ESI-MS(m / z):[M+H] + =400.1(calcd:400.0).

[0094] (Preparation of Hydrochloride Salt 32 of Compound 17) [ka]

[0095] Using compound 17 as a starting material, compound 32 could be prepared in 88% yield by following the preparation method of the hydrochloride salt of compound 10. 1 H NMR(500MHz,CD3OD):δ4.44-4.40(m,1H,H6),3.38-3.33(m,1H),3.26-3.22(m,1H),2.94-2.90(m,1H,H 11), 2.88 (s, 6H), 2.26-1.98 (m, 2H), 1.96-1.92 (m, 1H), 1.84-1.80 (m, 1H), 1.78-1.75 (m, 1H), 1.58-1.41 (m, 5H), 1.25-1.20 (m, 1H), 1.13-1.08 (m, 1H), 1.01-0.98 (m, 1H), 0.70-0.68 (m, 1H), 0.46-0.42 (m, 1H), 0.33-0.18 (m, 4H), 0.15-0.11 (m, 2H). The ROESY spectrum shows H6 and H 11 The signal correlation was confirmed, and the 11-H was found to be in the β-configuration. ESI-MS (m / z): [M+H] + =340.8(calcd:340.9).

[0096] Example 5: Compound 25 was converted to Compound 1 in plasma and HEPES (Experimental Method) Preparation of HEPES 7.4 solution: 1.6 g of NaCl, 0.074 g of KCl, 0.027 g of NaHPO, 0.2 g of glucose, and 1 g of 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) solution were added to 90 mL of distilled water, the pH was adjusted to 7.4 with 0.5 M NaOH, and the solution was made up to 100 mL with distilled water.

[0097] Plasma preparation: Mouse plasma was collected in an EP tube containing sodium heparin in advance, centrifuged at 8000 rpm at 4°C for 10 minutes, and the supernatant was used.

[0098] Sample analysis: 0.6 mg of compound 25 was dissolved in 250 μL of deionized water. 250 μL of mouse serum or HEPES 7.4 solution was added to the sample and incubated at 37 °C. Samples were taken at different time points. 20 μL of the sample was transferred to an EP tube, 60 μL of methanol was added, vortexed, and centrifuged at 12,000 rpm at 4 °C for 10 minutes. After 1 hour, 2 hours, 4 hours, 8 hours, and 12 hours, the supernatant was used for HPLC analysis. 10 μL of the sample was sampled, and the corresponding peak area was recorded. Chromatography conditions were as follows: column: Hanbang C18 (4.6 × 250 mm, 5 μm); mobile phase: acetonitrile: 10 mmol / mL ammonium formate solution = 60:40; flow rate: 1.0 mL / min; detection wavelength: 210 nm; column temperature: 30 °C.

[0099] (Experimental results) As shown in Figure 1, the content of compound 1 in the HEPES buffer solution increased at 1, 2, 4, 8, and 12 hours, with the contents being 5.36%, 11.05%, 19.64%, 39.29%, and 55.36%, respectively. This experimental result indicates that compound 25 can be converted to unchanged compound 1 as a prodrug in the HEPES buffer solution. As shown in Figure 2, the content of compound 1 in mouse plasma increased at 1, 2, 4, 8, and 12 hours, with the contents being 5.77%, 11.05%, 18.86%, 39.42%, and 55.88%, respectively. This experimental result indicates that compound 25 can be converted to unchanged compound 1 as a prodrug in mouse plasma.

[0100] [ka]

[0101] Similarly, other prodrug compounds can be converted to the corresponding drug substance compounds in plasma and HEPES.

[0102] Example 6: Water solubility test of prototype and its prodrug 20 μg of each of compounds 1 to 9 and 10 mg of each of compounds 24 to 32 were weighed out and added to 1 mL of deionized water and completely dissolved by sonication. After preparing a saturated solution and filtering, the solution was injected into an HPLC for analysis. The sample volumes were 1 μL, 3 μL, 5 μL, 10 μL, 15 μL, and 20 μL, respectively, and calibration curves for the corresponding compounds were prepared.

[0103] The unsaturated solution of the above compounds was prepared, and dissolved by ultrasonication for 4 hours. The solution was then placed in a water bath at 37°C and allowed to stand for 1 hour. The resulting unsaturated solution was centrifuged, and 30 μL of the supernatant was removed and diluted with 200 μL of deionized water. After filtration, the sample was analyzed by HPLC. The correlation data was substituted into the calibration curve obtained above to obtain the solubilities of the test compounds 1 to 9 and 24 to 32.

[0104] [Table 2]

[0105] As shown in Table 1, prodrug salts 24 to 32 had at least 100-fold improved water solubility compared to Arglabin, Dehydrocostus lactone, and the corresponding drug substances.

[0106] Example 7: Comparison of Pharmacokinetic Properties of Equimolar Amounts of Compounds 1 and 25 (Experimental materials) (Experimental Reagents) The drug of the present invention, prepared in Example 1 above; tolbutamide (internal standard, IS), Dalian Meilun Bio-Technology Co., Ltd.; dimethyl sulfoxide, Shanghai Taitan Science and Technology Co., Ltd.; normal saline, Chenxin Pharmaceutical Co., Ltd.; sodium carboxymethylcellulose, Aladdin Pharmaceuticals; methanol, acetonitrile and formic acid, Merck & Co.; pure water, Hangzhou Wahaha Group Co., Ltd.

[0107] (Experimental equipment) Refrigerated centrifuge, Eppendorf; vortex mixer, Scientific Industries; numerically controlled ultrasonic cleaner, Kunshan Ultrasonic Equipment Co., Ltd.; electronic balance, Sartorius; magnetic stirrer, IKA; electronic balance, Changzhou Lucky Electronic Equipment Co., Ltd.; H-Class / Xevo TQ-S micro liquid chromatography mass spectrometer, Waters.

[0108] (Laboratory animals) SPF-grade male SD rats weighing 200±20g were provided by Qinglongshan Animal Farm in Jiangning District, Nanjing. After purchase, they were housed at an ambient temperature of 23-26°C and humidity of 40-60% for 7 days, with free access to food and water. Animal production permit number: SCXK(Zhejiang)2019-0002.

[0109] (Experimental Method) (Establishment of UPLC-MS / MS measurement method) Chromatography conditions: Waters Acquity UPLC® BEH C 18 A 2.1 × 50 mm, 1.7 μm column was used; the mobile phase A was 0.1% formic acid in water, and the mobile phase B was acetonitrile. The gradient was: 0–1.0 min, 5%–30% B; 1.0–2.0 min, 30%–80% B; 2.0–3.0 min, 80%–80% B; 3.0–4.0 min, 80%–5% B; 4.0–5.0 min, 5%–5% B; total run time: 5 min; flow rate: 0.3 mL / min; column temperature: 30 °C; sampling volume: 2 μL.

[0110] Mass spectrometry conditions: An electrospray ion source (ESI) was used in positive ion monitoring mode, with a multiple reaction monitoring (MRM) scan. The ions used for detection were m / z 263.1 → 227.2 (compound 1), m / z 308.2 → 116.0 (compound 24), and m / z 270.9 → 91.0 (IS). MS operating parameters were set as follows: capillary voltage 1000 V, desolvation temperature 600 °C, and desolvation flow rate 1000 L / hr. The cone voltages for compounds 1, 25, and IS were 26 V, 48 V, and 14 V, respectively, and the collision energies were 44 V, 18 V, and 30 V, respectively. Data collection and analysis were performed using Masslynx 4.2.

[0111] (Plasma sample processing) After methodological considerations, plasma samples were pretreated using a 1:3 protein precipitation method, with methanol selected as the protein precipitant. 50 μL of rat plasma sample was aspirated, 150 μL of internal standard methanol solution (0.67 ng / mL) was added, and the mixture was centrifuged at 14,000 rpm at 4°C for 10 minutes. 2 μL of the supernatant was injected and analyzed by LC-MS / MS.

[0112] (Pharmacokinetic studies) A total of 24 male SD rats were randomly divided into two groups, one for oral gavage and one for tail vein injection (compound 1 and compound 25), with six rats in each group. Equimolar amounts of compound 1 (0.345 mmol / kg) and compound 25 (0.345 mmol / kg) were orally administered to the rats using 0.5% sodium carboxymethylcellulose (containing 10% DMSO) as a vehicle, and equimolar amounts of compound 1 (0.024 mmol / kg) and compound 25 (0.024 mmol / kg) were intravenously administered to the rats using saline (containing 5% DMSO) as a vehicle. After oral gavage, rats were bled via the orbit at 0.167, 0.25, 0.5, 0.75, 1, 1.5, 2, 4, 5, 6, 7, 8, 10, 12, and 24 hours post-dose. After tail vein injection, rats were bled via the orbit at 0.033, 0.083, 0.167, 0.25, 0.5, 0.75, 1, 1.5, 2, 4, 6, 8, 10, 12, and 24 hours post-dose. Whole blood collected via the orbital venous plexus was placed in a 1.5 mL EP tube pretreated with heparin sodium solution and centrifuged at 8000 rpm at 4°C for 10 minutes. Plasma was obtained and stored at -20°C until use.

[0113] (Data Analysis) The pharmacokinetic parameters were the elimination half-life (t 1 / 2 ), area under the concentration-time curve (AUC), mean residence time (MRT), apparent volume of distribution (V z / F ) and plasma elimination rate (CL z / F ) and calculated by the non-compartmental analysis model in DAS (Drugs and Statistics, version 3.0) software. The maximum concentration (C max ) and the time to reach the maximum concentration (T max ) were measured. All data are expressed as mean ± standard deviation (SD).

[0114] (Experimental results) As shown in Table 3, equimolar amounts of compound 1 and compound 25 were administered orally by gavage. The AUC value of compound 1 in the blood after oral gavage of 25 was nearly twice the AUC value of compound 1 in the blood after oral gavage of 1, and the C value of compound 1 in the blood after oral gavage of 25 was max The C value is the C of compound 1 in the blood after oral gavage of 1.max This was 10-fold higher than the original value, indicating that the prodrug significantly improved the oral absorption of Compound 1.

[0115] [Table 3]

[0116] Example 8: Activity test of compounds inhibiting NLRP3 inflammasome activation NLRP3 is an important pattern recognition receptor that can form the NLRP3 inflammasome via the adaptor protein ASC and pro-caspase-1. After activation, the NLRP3 inflammasome mediates caspase-1 activation, further promoting the maturation and secretion of IL-1β. To confirm whether the prepared guaiane-based sesquiterpene lactones 1–9 can inhibit NLRP3 inflammasome activation, we induced NLRP3 inflammasome activation with LPS and ATP and observed the effects of compounds 1–9 on IL-1β levels following NLRP3 inflammasome activation.

[0117] (Experimental materials) (Experimental Reagents) The drug of the present invention, prepared in Example 1 above; lipopolysaccharide (LPS) and adenosine triphosphate (ATP), Sigma; recombinant mouse macrophage colony stimulating factor (rmM-CSF), PeproTech; RPMI1640 medium, DMEM medium, and fetal bovine serum (FBS).

[0118] (Laboratory animals) C57BL / 6 mice, female, 6–8 weeks old, weighing 18–20 g, were provided by Qinglongshan Animal Farm, Jiangning District, Nanjing. Production permit number: SCXK(Su)2017-0001.

[0119] (Experimental Method) Isolation and culture of mouse bone marrow-derived macrophages (BMDMs) C57BL / 6 mice were sacrificed by cervical dislocation and immersed in 75% alcohol for 5–10 minutes. The two hind legs were then removed with scissors. The femur was removed and the foot bones were washed three times with PBS. A 1 mL syringe was filled with serum-free RPMI 1640 medium, the bones were clamped with tweezers, and cut open at both ends with scissors. The bone marrow was then sprayed into a 15 mL centrifuge tube with the syringe, repeatedly until the bone marrow was removed and the bone turned from red to white. The tube was then centrifuged at 1500 rpm for 55 minutes, the supernatant discarded, and the tube was resuspended in 1 mL of erythrocyte lysis solution. After repeated blowing, the tube was left to stand for 7 minutes to lyse the red blood cells. The tube was then centrifuged at 1500 rpm for 5 minutes, the supernatant discarded, and the tube was resuspended in RPMI 1640 medium containing 100 ng / mL rmM-CSF. The tube was then transferred to a 6-well plate for culture. After 6 to 7 days, the cells were observed and found to have a long spindle shape, indicating that they were in good condition and could be used for subsequent experiments.

[0120] (Establishment of a model for NLRP3 inflammasome activation) DMEM medium containing 1% FBS and 100 ng / mL LPS was prepared and the BMDMs in the 6-well plate were pre-stimulated with the medium for 3 hours. Arglabin (1, 3, 10, 30, 60, or 120 nM) was added and incubated for 1 hour, followed by ATP (5 mM) stimulation for 1 hour. The supernatant was collected in a 1.5 mL EP tube and subsequently used for IL-1β level detection.

[0121] [Table 4]

[0122] We induced an NLRP3 inflammasome activation model in BMDMs using LPS and ATP, and examined the effects of compounds 1–9 and 24–32 on IL-1β expression levels. As shown in Table 4, derivatives 1–9 all exhibited excellent inhibition of IL-1β activity, similar to the positive compound Arglabin, although the activity of the prodrug was slightly reduced.

[0123] Example 9: Anti-ulcerative colitis activity test of Compound 1 and its dimethyl fumarate prodrug 25 (Experimental Reagents) The drug of the present invention, prepared in Example 1 above; dextran sulfate sodium (DSS), MP Biomedicals; 5-aminosalicylic acid (5-ASA), Ethypharm Pharmaceuticals, France; carboxymethylcellulose sodium (CMC-Na), Xilonghua Plant, Shantou City, Guangdong; myeloperoxidase (MPO) kit, Nanjing Jiancheng Bioengineering Institute; o-toluidine, Shanghai Jingpun Biochemical Technology Co., Ltd.; hydrogen peroxide (H2O2) and glacial acetic acid, Nanjing Chemical Reagent Co., Ltd.

[0124] (Laboratory animals) Female C57BL / 6 mice, 6-8 weeks old and weighing 18-20 g, were provided by the Qinglongshan Animal Farm in Jiangning District, Nanjing. Production permit number: SCXK(Su)2017-0001. The animals had free access to food and water and were fed a standard granular diet. The room temperature was 22±2°C and humidity was 45±10%. After 3 days of adaptation, they were used in the experiments.

[0125] (Experimental Method) (Establishment of a mouse colitis model and group administration) Mice were randomly divided into seven groups, one of which was designated the normal group, and the remaining six groups were designated the model group, Arglabin (20 nmol / kg), Compound 1 (20 and 40 nmol / kg), and Compound 25 (20 and 40 nmol / kg) groups, each consisting of six mice. Except for the normal group, the remaining mice were given 2.5% DSS ad libitum for 7 days, then switched to simple distilled water for 3 days to establish the UC model. Starting on day 1 of model construction, Arglabin (20 nmol / kg / d), Compound 1 (20 and 40 nmol / kg / d), and Compound 25 (20 and 40 nmol / kg / d) were orally administered by gavage for 10 consecutive days. Mice in the normal and model groups were orally administered an equal volume of the vehicle, 0.5% CMC-Na, by gavage.

[0126] (Disease Activity Index score) The general living conditions of the mice in each group were observed, and the disease activity index (DAI) was evaluated. Specific daily observation parameters were the mouse weight, stool characteristics, and occult blood status. The scores for weight loss, stool characteristics, and occult blood status were then averaged to obtain a DAI score for each mouse, i.e., DAI = (weight loss score + stool characteristics score + occult blood score) / 3, which was used to evaluate disease activity.

[0127] As shown in Table 5, the evaluation criteria are as follows:

[0128] [Table 5]

[0129] Occult blood test method: Using the o-toluidine method, take a small amount of feces with a cotton swab onto a 24-well plate, first add 200 μL of o-toluidine glacial acetic acid solution, and then immediately add 200 μL of 3% hydrogen peroxide solution. If the color turns blue-green within 2 minutes, it is positive.

[0130] (Sample collection) One hour after the final administration, blood was collected from the venous plexus at the back of the eye, left to stand at room temperature for 2 hours, and centrifuged at 3000 rpm for 20 minutes. The serum was aspirated, aliquoted, and frozen at -70°C. After blood collection was complete, the mice were sacrificed by cervical dislocation, the abdominal cavity was opened, and the colon tissue was removed 1 cm from the anus, washed twice with pre-cooled PBS, and frozen at -70°C.

[0131] (Colon length measurement) Colon shortening is one of the main characteristics of DSS-induced UC model mice. UC mice were intraperitoneally dissected, and the colon and distal ileum tissues were isolated. The changes in the external morphology of the colon tissue were observed, and its length was measured, recorded, and photographed.

[0132] (MPO activity measurement) 40 mg of colonic tissue was collected from each group of mice, and 400 μL of pre-chilled saline was added to prepare a tissue homogenate. After centrifugation at 1200 rpm at 4°C for 5 minutes, the supernatant was collected. Various reagents were added sequentially according to the kit instructions. Finally, the absorbance of each well was measured at 460 nm using a microplate reader, and MPO activity was calculated. MPO activity (U / g tissue) = (OD value of measurement tube - OD value of control tube) / 11.3 × sampling amount (g)

[0133] (Data Statistics) All data are expressed as mean ± SEM. Analysis of variance was used to analyze the significance of the differences. For those with significant differences in analysis of variance, one-way ANOVA and Dunnett's were used to further verify the differences between the comparison groups. P values ​​less than 0.05 were considered significant.

[0134] (Experimental results) Mice in the normal group showed normal activity and defecation, and gradually increased in weight. Mice administered DSS showed symptoms such as loose stools and semi-formed, non-adherent stools over the course of the experiment, decreased in activity, and showed a significant weight loss. Disease activity indicators were evaluated by daily observation of the mice for weight loss, stool quality, and stool blood status. As shown in Table 6, compared with the DSS group, compound 25 (40 nmol / kg) significantly reduced the DSS-induced increase in colitis DAI score, and the activity of prodrug 25 at equivalent doses was significantly stronger than that of drug substance 1.

[0135] (Impact on DAI score) [Table 6]

[0136] (Effect on colon length) [Table 7]

[0137] As shown in Table 7, the colon length of the mice in the DSS group was significantly shortened compared with that of the normal group. Oral administration of compound 25 (40 nmol / kg) by gavage significantly inhibited the shortening of the colon length of the mice, and the activity of prodrug 25 at the same dose was clearly stronger than that of drug substance 1.

[0138] (Effects on colonic tissue MPO activity) Granulocytes can synthesize myeloperoxidase (MPO) in the bone marrow before entering the blood circulation, and the latter is stored in azurophilic granules. MPO can kill pathogenic microorganisms by generating hypochlorous acid and regulate inflammatory responses. MPO accounts for approximately 5% of the dry weight of cells, and this characteristic can be used to measure the number of neutrophils in tissues. As shown in Table 8, colonic tissue MPO activity in mice treated with DSS was significantly elevated compared with that in normal mice. Oral gavage of 25 (40 nmol / kg) significantly reduced MPO activity in colonic tissues of mice, and the activity of the prodrug 25 at equivalent doses was significantly stronger than that of the original drug 1.

[0139] [Table 8]

[0140] Example 10: Anti-acute gouty arthritis activity test of Compound 25 (Experimental Reagents) The drug of the present invention, prepared in Example 2 above; colchicine tablets, Xisapanna Pharmaceutical Co., Ltd.; prednisolone acetate, Tianjin Xinyijin Pharmaceutical Co., Ltd.; carboxymethylcellulose sodium (CMC-Na), China National Pharmaceutical Group Chemical Reagent Co., Ltd.; pentobarbital sodium, China National Pharmaceutical Group Shanghai Chemical Reagent Co., Ltd.; sodium urate (MSU), Sigma-Aldrich; IL-1β radioimmunoassay kit, IL-18 radioimmunoassay kit, Beijing Huaying Institute of Biotechnology.

[0141] (Laboratory animals) SPF-level SD rats, female, weighing 200-220 g, were provided by Hangzhou Medical University. Production permit number: SCXK (Zhejiang) 2019-0002. Animal use permit number: SYXK (Suzhou) 2019-0004. The animals had free access to food and water and were fed a standard granular diet. The room temperature was 22±2°C, and the humidity was 45±10%. After acclimatization for 3 days, they were used in the experiment.

[0142] (Experimental Method) (Rat acute gouty arthritis model) During the adaptation period, rats were anesthetized intraperitoneally with 2% pentobarbital sodium at a dose of 0.25 mL / 100 g, and the weekly diameter of the ankle joint of the left hind limb of each rat was measured. After the adaptation period, rats were anesthetized intraperitoneally with 2% pentobarbital sodium at a dose of 0.25 mL / 100 g. After anesthesia, eight rats were randomly selected as the blank group and injected with saline into the left hind limb joint cavity. The remaining rats were injected with 200 μL of sodium urate into the left hind limb joint cavity to construct the model. Six hours after model construction, the rats were anesthetized with pentobarbital sodium, and the joint circumference was measured using a 2 mm-wide paper tape. The rats were divided into seven groups based on the degree of joint swelling. The groups were blank (equal volume of CMC-Na); model and positive control (equal volume of CMC-Na); colchicine (0.5 mg / kg); prednisolone (3.125 mg / kg); three groups administered low, medium, and high doses of Compound 25 (1.5, 5.0, and 15 mg / kg); 8 mice in the blank group; and 10 mice in each of the remaining groups.

[0143] (Administration by group) After the model was established, the rats were administered once at 10.5 and 22.5 hours. The rats in the blank and model groups were administered CMC-Na (0.5%) by oral gavage, the rats in the colchicine group were administered colchicine solution by oral gavage, the rats in the prednisolone group were administered prednisolone solution by oral gavage, and the rats in each dose group were administered different concentrations of Compound 25 by oral gavage, with the administration volume being 10 mL / kg.

[0144] (Joint swelling assessment) Joint swelling is a key feature of the MSU-induced acute gouty arthritis model in rats. The circumference of the left hind ankle joint was measured before and 6, 12, and 24 hours after model construction, and the joint swelling rate was calculated. The weekly diameter at a point 0.5 mm below the left hind ankle joint of each group of rats was measured twice using a 2 mm paper tape and a straightedge. The average value was then used to calculate the joint swelling rate. Joint swelling rate = (joint circumference at the time of measurement - initial joint circumference) / initial joint circumference

[0145] (Data Statistics) All data are expressed as mean ± SEM. Analysis of variance was used to analyze the significance of the differences. For those with significant differences in analysis of variance, one-way ANOVA and Dunnett's were used to further verify the differences between the comparison groups. P values ​​less than 0.05 were considered significant.

[0146] (Experimental results) [Table 9]

[0147] (Effect on MSU-induced joint swelling rate) As shown in Table 9, the ankle swelling rates of rats in the model group 6 hours, 12 hours, and 24 hours after model construction were all significantly higher than those of the control group. The ankle swelling rates of rats in each administration group 1.5 hours after the first administration (12 hours after model construction) were all reduced, but not significantly. The ankle swelling rates of rats in each administration group 1.5 hours after the second administration (24 hours after model construction) were all significantly reduced, suggesting that compound 25 significantly reduces the ankle swelling rate of rats with MSU-induced acute gouty arthritis.

[0148] (Addendum) (Appendix 1) A guaiane-based sesquiterpene derivative represented by general formula I or a pharmaceutically acceptable salt thereof, [ka] R1 and R2 together form a double bond, or R1 is hydrogen or deuterium and R2 is [ka] wherein R3 and R4 are each hydrogen, an alkyl group, or a cycloalkyl group, or R3, R4 and the N atom form a 3- to 9-membered ring structure, and the ring contains an alkyl group, an ester group, an aryl group, an alkylaryl group, an arylalkyl group, an arylalkenyl group, an arylalkynyl group, or a heterocyclic substituent; R5 and R6 are bonded together by a single bond to form a cyclopropane, or when R5 and R6 do not form a cyclopropane, R5 is a methyl group and R6 is a hydroxyl group, an alkoxy group, an ester group, a halogen atom, or forms a double bond with the carbon atom in the ortho position; R7 is hydrogen or a hydroxyl group; R8 and R9 are joined by a single bond to form a cyclopropane, and R 10 is hydrogen, or R8 is a methyl group and R9 is R 10 to form a cyclopropane, A guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof.

[0149] (Appendix 2) The following compound: [ka] [ka] 2. The guaiane-type sesquiterpene derivative or a pharmaceutically acceptable salt thereof according to claim 1, wherein the guaiane-type sesquiterpene derivative or the pharmaceutically acceptable salt thereof is selected from the group consisting of:

[0150] (Appendix 3) 2. The guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof according to claim 1, wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, sulfate, phosphate, maleate, fumarate, and citrate.

[0151] (Appendix 4) 4. The guaiane-type sesquiterpene derivative or a pharmaceutically acceptable salt thereof according to claim 3, wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride and fumarate. [ka]

[0152] (Appendix 5) A pharmaceutical composition comprising the guaiane-type sesquiterpene derivative or a pharmaceutically acceptable salt thereof according to any one of appendices 1 to 4, and a pharmaceutically acceptable carrier.

[0153] (Appendix 6) Use of the guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof according to any one of Appendices 1 to 4, and the pharmaceutical composition according to Appendices 5, in the manufacture of a drug for treating an NLRP3 inflammasome-associated disease.

[0154] (Appendix 7) The use described in Appendix 6, characterized in that the NLRP3 inflammasome-related disease includes immune diseases, autoimmune diseases, malignant tumors, skin diseases, cardiovascular diseases, liver-related diseases, renal system-related diseases, gastrointestinal tract-related diseases, central nervous system diseases, metabolic diseases, endocrine-related diseases, respiratory diseases, lymphatic system diseases, inflammation, infectious diseases, eye diseases, psychiatric diseases, and pain.

[0155] (Appendix 8) The NLRP3 inflammasome-related diseases include (1) cryopyrin protein-associated periodic syndromes (CAPS): Muckle-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), and chronic infantile neurological cutaneous and articular syndrome (NOMID); and (2) autoinflammatory diseases: familial Mediterranean fever (FMF), TNF receptor-associated periodic syndrome (TRAPS), mevalonate kinase deficiency (MKD), hyperimmunoglobulin D and periodic fever syndrome (HIDS), interleukin-1 receptor DIRA deficiency, Majeed syndrome, septic arthritis, pyoderma gangrenosum and acne (PAPA), and A2. (1) Haploinsufficiency HA20, childhood granulomatous arthritis PGA, PLCG2-related antibody deficiency & immune dysregulation PLAID, PLCG2-related autoinflammation, antibody deficiency & immune dysregulation APLAID, and sideroblastic anemia with B-cell immunodeficiency, periodic fever, and growth retardation SIFD; (2) Sweet's syndrome: chronic nonbacterial osteomyelitis CNO, chronic recurrent multifocal osteomyelitis CRMO, synovitis, acne, impetigo, hyperostosis & osteitis syndrome SAPHO; (3) Autoimmune diseases: multiple sclerosis MS, type 1 diabetes, psoriasis, rheumatoid arthritis, Behcet's disease, Sjogren's disease (5) Respiratory diseases: chronic obstructive pulmonary disease (COPD), steroid-resistant asthma, asbestosis, silicosis, and cystic fibrosis; (6) Central nervous system diseases: Parkinson's disease, Alzheimer's disease, motor neuron disease, Huntington's disease, cerebral malaria, and brain damage due to pneumococcal meningitis; (7) Metabolic diseases: type 2 diabetes, atherosclerosis, obesity, gout, and pseudogout; (8) Eye diseases: ocular epidermis, age-related macular degeneration (AMD), corneal infection, uveitis, and dry eye; (9) Kidney-related diseases: chronic kidney disease, silicosis, and cystic fibrosis. (10) Liver-related diseases: nonalcoholic steatohepatitis and alcoholic liver disease; (11) Skin-related inflammatory reactions: contact allergy and sunburn; (12) Joint-related inflammatory reactions: osteoarticular, systemic juvenile idiopathic arthritis, adult Still's disease, relapsing polychondritis; (13) Viral infections: dengue virus, Zika virus, influenza, AIDS virus; (14) Hidradenitis suppurativa (HS) and other skin diseases causing cysts; (15) Cancer: lung cancer, pancreatic cancer, gastric cancer, myelodysplastic syndrome, abdominal aortic aneurysm, and leukemia;(16) The use according to Appendix 6, including polymyositis, colitis, pericarditis, helminth infections, bacterial infections, wound healing, depression, stroke, myocardial infarction, hypertension, Dressler's syndrome, and ischemia-reperfusion injury;

[0156] (Appendix 9) The use of claim 8, wherein the colitis includes ulcerative colitis.

[0157] (Appendix 10) The use described in Appendix 6, wherein the NLRP3 inflammasome-associated disease includes acute gouty arthritis.

Claims

1. A guaiane-based sesquiterpene derivative represented by general formula I or a pharmaceutically acceptable salt thereof, 【Chemical 1】 R １ , R ２ together form a double bond, or R １ is hydrogen or deuterium, R ２ teeth, 【Chemistry 2】 Among them, R ３ and R ４ are each an alkyl group, or R ３ , R ４ and the N atom form a 5- to 6-membered ring structure, the ring being a ring structure selected from tetrahydropyrrole, piperidine, piperazine, and morpholine; R ５ is a methyl group, and R ６ is a hydroxyl group, an alkoxy group, an ester group, a halogen atom, or forms a double bond with the carbon atom at the ortho position, R ７ is hydrogen or a hydroxyl group, R ８ is a methyl group, and R ９ is R １０ to form a cyclopropane, A guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof.

2. The following compound: 【Chemistry 3】 【Chemistry 4】 2. The guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof according to claim 1, wherein the guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

3. The following compound: 【Chemistry 5】 2. The guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof according to claim 1, wherein the guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

4. 2. The guaiane-based sesquiterpene derivative or its pharmaceutically acceptable salt according to claim 1, wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, sulfate, phosphate, maleate, fumarate, and citrate.

5. the pharmaceutically acceptable salt is a hydrochloride or a fumarate; 【Chemistry 6】 5. The guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof according to claim 4, wherein the guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

6. A pharmaceutical composition comprising the guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, and a pharmaceutically acceptable carrier.

7. Use of the guaiane-based sesquiterpene derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 in the manufacture of a drug for treating an NLRP3 inflammasome-associated disease.

8. Use of the pharmaceutical composition of claim 6 in the manufacture of a medicament for treating an NLRP3 inflammasome-associated disease.

9. The NLRP3 inflammasome-related diseases include: (1) cryopyrin protein-associated periodic syndromes (CAPS): Muckle-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), and chronic infantile neurological, cutaneous, and articular syndrome (NOMID); and (2) autoinflammatory diseases: familial Mediterranean fever (FMF), TNF receptor-associated periodic syndrome (TRAPS), mevalonate kinase deficiency (MKD), hyperimmunoglobulin D and periodic fever syndrome (HIDS), interleukin-1 receptor (DIRA) deficiency, Majeed syndrome, septic arthritis, pyoderma gangrenosum / acne (PAPA), and A2. (1) Haploinsufficiency syndrome HA20, childhood granulomatous arthritis PGA, PLCG2-related antibody deficiency & immune dysregulation PLAID, PLCG2-related autoinflammation, antibody deficiency & immune dysregulation APLAID, and sideroblastic anemia with B-cell immunodeficiency, periodic fever, and growth retardation SIFD; (2) Sweet's syndrome: chronic nonbacterial osteomyelitis CNO, chronic recurrent multifocal osteomyelitis CRMO, synovitis, acne, impetigo, hyperostosis & osteitis syndrome SAPHO; (3) Autoimmune diseases: multiple sclerosis MS, type 1 diabetes, psoriasis, rheumatoid arthritis, Behcet's disease, Sjogren's disease (5) Respiratory diseases: chronic obstructive pulmonary disease (COPD), steroid-resistant asthma, asbestosis, silicosis, and cystic fibrosis; (6) Central nervous system diseases: Parkinson's disease, Alzheimer's disease, motor neuron disease, Huntington's disease, cerebral malaria, and brain damage due to pneumococcal meningitis; (7) Metabolic diseases: type 2 diabetes, atherosclerosis, obesity, gout, pseudogout; (8) Eye diseases: ocular epidermis, age-related macular degeneration (AMD), corneal infection, uveitis, and dry eye; (9) Kidney-related diseases: chronic kidney disease, silicosis, and cystic fibrosis. (10) Liver-related diseases: nonalcoholic steatohepatitis and alcoholic liver disease; (11) Skin-related inflammatory reactions: contact allergies and sunburn; (12) Joint-related inflammatory reactions: osteoarticular, systemic juvenile idiopathic arthritis, adult Still's disease, relapsing polychondritis; (13) Viral infections: dengue virus, Zika virus, influenza, AIDS virus; (14) Hidradenitis suppurativa (HS) and other skin diseases causing cysts; (15) Cancer: lung cancer, pancreatic cancer, gastric cancer, myelodysplastic syndrome, abdominal aortic aneurysm, and leukemia;(16) polymyositis, colitis, pericarditis, helminth infection, bacterial infection, wound healing, depression, stroke, myocardial infarction, hypertension, Dressler's syndrome, ischemia-reperfusion injury; (17) lymphatic system disease; (18) psychiatric disorder; (19) pain.

10. The NLRP3 inflammasome-related diseases include: (1) cryopyrin protein-associated periodic syndromes (CAPS): Muckle-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), and chronic infantile neurological, cutaneous, and articular syndrome (NOMID); and (2) autoinflammatory diseases: familial Mediterranean fever (FMF), TNF receptor-associated periodic syndrome (TRAPS), mevalonate kinase deficiency (MKD), hyperimmunoglobulin D and periodic fever syndrome (HIDS), interleukin-1 receptor (DIRA) deficiency, Majeed syndrome, septic arthritis, pyoderma gangrenosum / acne (PAPA), and A2. (1) Haploinsufficiency syndrome HA20, childhood granulomatous arthritis PGA, PLCG2-related antibody deficiency & immune dysregulation PLAID, PLCG2-related autoinflammation, antibody deficiency & immune dysregulation APLAID, and sideroblastic anemia with B-cell immunodeficiency, periodic fever, and growth retardation SIFD; (2) Sweet's syndrome: chronic nonbacterial osteomyelitis CNO, chronic recurrent multifocal osteomyelitis CRMO, synovitis, acne, impetigo, hyperostosis & osteitis syndrome SAPHO; (3) Autoimmune diseases: multiple sclerosis MS, type 1 diabetes, psoriasis, rheumatoid arthritis, Behcet's disease, Sjogren's disease (5) Respiratory diseases: chronic obstructive pulmonary disease (COPD), steroid-resistant asthma, asbestosis, silicosis, and cystic fibrosis; (6) Central nervous system diseases: Parkinson's disease, Alzheimer's disease, motor neuron disease, Huntington's disease, cerebral malaria, and brain damage due to pneumococcal meningitis; (7) Metabolic diseases: type 2 diabetes, atherosclerosis, obesity, gout, pseudogout; (8) Eye diseases: ocular epidermis, age-related macular degeneration (AMD), corneal infection, uveitis, and dry eye; (9) Kidney-related diseases: chronic kidney disease, silicosis, and cystic fibrosis. (10) Liver-related diseases: nonalcoholic steatohepatitis and alcoholic liver disease; (11) Skin-related inflammatory reactions: contact allergies and sunburn; (12) Joint-related inflammatory reactions: osteoarticular, systemic juvenile idiopathic arthritis, adult Still's disease, relapsing polychondritis; (13) Viral infections: dengue virus, Zika virus, influenza, AIDS virus; (14) Hidradenitis suppurativa (HS) and other skin diseases causing cysts; (15) Cancer: lung cancer, pancreatic cancer, gastric cancer, myelodysplastic syndrome, abdominal aortic aneurysm, and leukemia;(16) polymyositis, colitis, pericarditis, helminth infection, bacterial infection, wound healing, depression, stroke, myocardial infarction, hypertension, Dressler's syndrome, ischemia-reperfusion injury; (17) lymphatic system disease; (18) psychiatric disorder; (19) pain.

11. 10. The use according to claim 9, wherein the colitis is ulcerative colitis.

12. The use according to claim 7, wherein the NLRP3 inflammasome-associated disease is acute gouty arthritis.

13. The use according to claim 8, wherein the NLRP3 inflammasome-associated disease is acute gouty arthritis.

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