Dihydroartemisinin derivatives and uses thereof

Abstract

The present application relates to a kind of dihydroartemisinin derivatives of structure shown in formula (I) or its pharmaceutically acceptable salt or its stereoisomer, and its application.The dihydroartemisinin derivative synthesized in the present application can significantly inhibit the expression of NLRP3 inflammasome, inhibit the activation of NLRP3 pathway;In the cell model of NLRP3 inflammasome activation, this kind of dihydroartemisinin derivative can inhibit the secretion and release of IL-1β and TNF-α in concentration-dependent manner, so as to be used for treating the disease related to NLRP3 inflammasome activation.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to dihydroartemisinin derivatives and their applications. Background Technology

[0002] The NLRP3 inflammasome is a NOD-like receptor composed of three parts: the inflammasome sensor molecule (NLRP3 protein), the adaptor protein ASC, and the effector molecule pro-caspase-1. It is a multi-protein complex located in the cytoplasm. Upon activation of the NLRP3 inflammasome, pro-caspase-1 self-cleaves into active caspase-1, which further cleaves pro-IL-1β and pro-IL-18 into active interleukin-1β (IL-1β) and interleukin-18 (IL-18), ultimately leading to an inflammatory response and pyroptosis. Extensive evidence suggests a close association between the NLRP3 inflammasome and many human diseases, such as Alzheimer's disease, gout, multiple sclerosis, type II diabetes, and inflammatory bowel disease.

[0003] Artemisinin derivatives possess a wide range of biological activities due to their unique peroxy-bridge structure. In addition to their classic antimalarial effects, existing studies (including some clinical explorations) have demonstrated their potential therapeutic value for autoimmune diseases such as lupus erythematosus, exhibiting certain immunosuppressive effects. However, existing artemisinin-based drugs have relatively weak inhibitory activity against the NLRP3 signaling pathway and suffer from poor bioavailability, limiting their application in a broader range of NLRP3-related inflammatory diseases. Therefore, developing novel artemisinin derivatives that can efficiently target the NLRP3 pathway has significant clinical translational implications. Summary of the Invention

[0004] Based on this, the purpose of this invention is to develop a novel artemisinin derivative that can efficiently target the NLRP3 pathway.

[0005] The technical solutions for achieving the above objectives include the following.

[0006] In a first aspect, the present invention provides a dihydroartemisinin derivative of formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

[0007]

[0008] Wherein, W is selected from: , , or ;

[0009] R is selected from: or ;

[0010] R 1 and R 2 Each and every one is independently selected from: C1-C6 alkyl groups;

[0011] n1 and n2 are independently selected from 0, 1, 2, 3 or 4, and n1 + n2 = 0~4;

[0012] m1 and m2 are each independently selected from: 1, 2, 3, 4 or 5, and m1 + m2 = 2~6;

[0013] y is selected from: 0, 1, 2 or 3.

[0014] Secondly, the present invention provides the use of the dihydroartemisinin derivatives described herein or pharmaceutically acceptable salts thereof or stereoisomers thereof in the preparation of NLRP3 inflammasome inhibitors.

[0015] Thirdly, the present invention provides the use of the dihydroartemisinin derivatives described herein or pharmaceutically acceptable salts thereof or stereoisomers thereof in the preparation of medicaments for the prevention and / or treatment of diseases associated with NLRP3 inflammasome activation.

[0016] Fourthly, the present invention provides a pharmaceutical composition for the prevention and / or treatment of diseases associated with NLRP3 inflammasome activation, prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient includes the dihydroartemisinin derivative of the present invention or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

[0017] The present invention has the following beneficial effects:

[0018] This invention synthesizes a novel class of dihydroartemisinin derivatives that can significantly inhibit the expression of the NLRP3 inflammasome and suppress the activation of the NLRP3 pathway. In a cell model of NLRP3 inflammasome activation, these dihydroartemisinin derivatives can inhibit the secretion and release of IL-1β and TNF-α in a concentration-dependent manner, thereby enabling their use in the treatment of diseases associated with NLRP3 inflammasome activation. Attached Figure Description

[0019] Figure 1 The effects of the compound on the viability of THP-1 and J774A.1 cells.

[0020] Figure 2 To investigate the effect of compound 21 on IL-1β secretion in a J774A.1 cell model activated by the NLRP3 inflammasome.

[0021] Figure 3To analyze the effect of compound 21 on the expression of pro-IL-1β and NLRP3 in a J774A.1 cell model activated by NLRP3 inflammasome using Western blotting.

[0022] Figure 4 The effect of compound 21 on LPS-mediated TNF-α release in J774A.1 cells. Detailed Implementation

[0023] To facilitate understanding of the present invention, a more complete description will be provided below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0024] Unless otherwise specified, experimental methods in the following examples are generally performed under standard conditions or as recommended by the manufacturer. All commonly used chemical reagents used in the examples are commercially available products.

[0025] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used in this invention includes any and all combinations of one or more of the associated listed items.

[0026] Furthermore, as used herein, the term "or" is an inclusive "or" sign and is equivalent to the term "and / or" unless the context clearly specifies otherwise. The term "based on" is not exclusive and allows for basing on other factors not described unless the context clearly specifies otherwise. Additionally, throughout the specification, the meanings of "an," "a," and "the" include plural indicators. The meaning of "in" includes both "in" and "on."

[0027] Some embodiments of the present invention involve dihydroartemisinin derivatives of formula (I) or pharmaceutically acceptable salts thereof or stereoisomers thereof.

[0028]

[0029] Wherein, W is selected from: , , or ;

[0030] R is selected from: or ;

[0031] R 1 and R 2 Each and every one is independently selected from: C1-C6 alkyl groups;

[0032] n1 and n2 are independently selected from 0, 1, 2, 3 or 4, and n1 + n2 = 0~4;

[0033] m1 and m2 are each independently selected from: 1, 2, 3, 4 or 5, and m1 + m2 = 2~6;

[0034] y is selected from: 0, 1, 2 or 3.

[0035] In the compounds of this invention, when any variable (e.g., R, etc.) appears more than once in any component, the definition of each occurrence is independent of the definition of other occurrences. Similarly, combinations of substituents and variables are permitted, provided such combinations stabilize the compound. A line drawn from a substituent into the ring system indicates that the bond referred to can be attached to any substituted ring atom. If the ring system is polycyclic, it means that such a bond is attached only to any suitable carbon atom of a neighboring ring. It will be understood that those skilled in the art can select the substituents and substitution patterns of the compounds of this invention to provide chemically stable compounds that can be readily synthesized from readily available starting materials using techniques in the art and the methods described below. If a substituent is itself substituted by more than one group, it should be understood that these groups can be on the same carbon atom or different carbon atoms, as long as structural stability is achieved.

[0036] As used herein, the term "alkyl" refers to both branched and straight-chain saturated aliphatic hydrocarbon groups having a specific number of carbon atoms. For example, the definition of "C1-C6 alkyl" includes groups having 1, 2, 3, 4, 5, or 6 carbon atoms arranged in a straight or branched chain. Specifically, "C1-C6 alkyl" includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, hexyl, etc.

[0037] In some implementations, n1+n2=1, 2 or 3.

[0038] In some implementations, m1+m2=2, 3 or 4.

[0039] In some implementations, W is selected from: , , , or .

[0040] In some implementations, R is selected from: or .

[0041] In some implementations, R 1 and R 2 Each is independently selected from: methyl, ethyl, n-propyl, and isopropyl.

[0042] In some implementations, R is selected from:

[0043] .

[0044] Some embodiments of the present invention also relate to the use of the dihydroartemisinin derivatives described herein or pharmaceutically acceptable salts thereof or stereoisomers thereof in the preparation of NLRP3 inflammasome inhibitors.

[0045] Some embodiments of the present invention also relate to the use of the dihydroartemisinin derivatives of the present invention or pharmaceutically acceptable salts thereof or stereoisomers thereof in the preparation of medicaments for the prevention and / or treatment of diseases associated with NLRP3 inflammasome activation.

[0046] Some embodiments of the present invention also relate to a pharmaceutical composition for the prevention and / or treatment of diseases associated with NLRP3 inflammasome activation, prepared from an active ingredient and pharmaceutically acceptable excipients, said active ingredient comprising the dihydroartemisinin derivatives of the present invention or pharmaceutically acceptable salts thereof or stereoisomers thereof.

[0047] The present invention will be further described in detail below with reference to specific embodiments.

[0048] The synthetic route of the compounds of this invention is as follows:

[0049]

[0050]

[0051] Example 1: Preparation of fumaric acid ((3R,5aS,6R,8aS,9R,10S,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isochromen-10-yl ethyl ester (compound 1)

[0052]

[0053] Dihydroartemisinin (DHA, 508.7 mg, 2.0 mmol) was dissolved in dichloromethane, and triethylamine (556.0 μL, 4.0 mmol) was added. The mixture was kept in an ice bath under nitrogen protection. Fumarate monoethyl ester chloride (400.0 μL, 3.0 mmol) was added, and the mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC, concentrated under reduced pressure, and purified by column chromatography to give compound 1, with a mass of 457.3 mg. The product was a yellow oily liquid, with a yield of 55.7%.1 H NMR (400 MHz, CDCl3) δ 6.98 – 6.86 (m, 2H), 5.86 (d, J =9.9 Hz, 1H), 5.47 (s, 1H), 4.26 (q, J = 7.1 Hz, 2H), 2.70 – 2.59 (m, 1H),2.43 – 2.33 (m, 1H), 2.09 – 2.00 (m, 1H), 1.94 – 1.86 (m, 1H), 1.82 – 1.70(m, 2H), 1.65 (dt, J = 13.8, 4.5 Hz, 1H), 1.52 – 1.46 (m, 1H), 1.43 (s, 3H),1.42 – 1.36 HRMS (ESI) calcd for C 21 H 30 O8 [M+Na] + 433.1833, found 433.1830.

[0054] Example 2: Preparation of 1-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isochromen-10-yl)piperazine (compound 2)

[0055]

[0056] Piperazine (2411.9 mg, 28.0 mmol) was dissolved in dichloromethane under nitrogen protection and stirred. DHA (1990.5 mg, 7.0 mmol) was dissolved in dichloromethane under nitrogen protection and stirred for 5 min. Then, 50 μL of anhydrous DMSO was added and stirred for 10 min. Oxaloyl chloride (651.6 μL, 7.7 mmol) was slowly added until no obvious bubbles were generated, and stirring was continued for 10 min. The resulting reaction solution containing DHA was slowly added to the dichloromethane solution of piperazine and stirred at room temperature for 12 h. The reaction progress was monitored by TLC. The product was washed three times each with saturated sodium bicarbonate aqueous solution and saturated brine. The organic phase was dried over anhydrous sodium sulfate. The silica gel column was first alkalized with ammonia before column chromatography purification to obtain compound 2, with a mass of 1487.8 mg. The product was a yellow oily liquid with a yield of 60.3%. 1H NMR (400 MHz, CDCl3) δ 5.27 (s, 1H), 3.98 (d, J =10.2 Hz, 1H), 3.03 – 2.99 (m, 1H), 2.97 (d, J = 2.7 Hz, 1H), 2.94 – 2.88 (m,3H), 2.71 – 2.65 (m, 2H), 2.61 – 2.55 (m, 1H), 2.43 (s, 2H), 2.39 – 2.28 (m,2H), 2.05 – 1.95 (m, 1H), 1.91 – 1.80 (m, 1H), 1.75 – 1.64 (m, 2H), 1.56 –1.43 (m, 2H), 1.39 HRMS (ESI) calcd for C 19 H 32 N₂O₄[M+H] + 353.2435, found 353.2435.

[0057] Example 3: Preparation of 1-(4-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isochromen-10-yl)piperazin-1-yl)prop-2-en-1-one (compound 3)

[0058]

[0059] Compound 2 (200.0 mg, 0.567 mmol) was dissolved in dichloromethane, and triethylamine (157.6 μL, 1.134 mmol) was added. The mixture was then placed in an ice bath under nitrogen protection. Acryloyl chloride (69.1 μL, 0.851 mmol) was added, and the mixture was stirred at room temperature for 4 h. The reaction progress was monitored by TLC. The reaction was quenched by stirring with methanol for 0.5 h, concentrated under reduced pressure, and purified by column chromatography to give compound 3, with a mass of 83.7 mg. The product was a yellow oily liquid, with a yield of 36.3%. 1H NMR (400 MHz, CDCl3) δ 6.56 (dd, J = 16.7,10.7 Hz, 1H), 6.26 (dd, J = 16.9, 2.1 Hz, 1H), 5.67 (dd, J = 10.7, 2.0 Hz,1H), 5.27 (s, 1H), 4.04 (d, J = 10.3 Hz, 1H), 3.67 (s, 2H), 3.61 – 3.46 (m,2H), 2.99 (dt, J = 11.3, 5.0 Hz, 2H), 2.72 – 2.64 (m, 2H), 2.63 – 2.54 (m,1H), 2.34 (td, J = 13.9, 3.8 Hz, 1H), 2.04 – 1.95 (m, 1H), 1.90 – 1.81 (m,1H), 1.74 – 1.67 (m, 2H), 1.58 – 1.40 (m, 3H), 1.38 (s, 3H), 1.37 – 1.29 (m,2H), 1.07 – 0.97 (m, 1H), 0.95 (d, J = 6.3 Hz, 3H), 0.82 (d, J = 7.2 Hz, 3H);HRMS (ESI) calcd for C 22 H 34 N₂O₅ [M+H] + 407.2541, found 407.2543.

[0060] Example 4: Preparation of 1-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isochromen-10-yl)-4-(vinylsulfonyl)piperazine (compound 4)

[0061]

[0062] Compound 2 (200.0 mg, 0.567 mmol) was dissolved in dichloromethane, and triethylamine (236.4 μL, 1.701 mmol) was added. The mixture was kept in an ice bath under nitrogen protection. 2-Chloroethanesulfonyl chloride (65.2 μL, 0.624 mmol) was added, and the mixture was stirred for 10 min. The mixture was then transferred to room temperature and stirred for another 4 h. The reaction was monitored by TLC. Methanol was added and stirred for 0.5 h to quench the reaction. The mixture was concentrated under reduced pressure and purified by column chromatography to give compound 4, with a mass of 95.9 mg. The product was a yellow oily liquid, with a yield of 38.2%. 1H NMR (400 MHz, CDCl3) δ 6.43 (dd, J = 16.6, 9.9 Hz, 1H), 6.23 (d, J = 16.6 Hz, 1H), 6.02 (d,J = 10.0 Hz, 1H), 5.26 (s, 1H), 4.03 (d, J = 10.2 Hz, 1H), 3.22 – 3.10 (m,4H), 3.09 – 3.02 (m, 2H), 2.82 – 2.72 (m, 2H), 2.62 – 2.49 (m, 1H), 2.40 –2.28 (m, 1H), 2.04 – 1.97 (m, 1H), 1.92 – 1.81 (m, 1H), 1.74 – 1.65 (m, 2H), 1.57 – 1.50 (m, 1H), 1.50 – 1.43 (m, 1H), 1.43 – 1.35 (m, 4H), 1.35 – 1.27(m, 2H), 1.06 – 0.97 (m, 1H), 0.94 (d, J = 6.2 Hz, 3H), 0.78 (d, J = 7.2 Hz, 3H); HRMS (ESI) calcd for C 21 H 34 N₂O₆S [M+H] + 443.2211, found 443.2205.

[0063] Example 5: Preparation of 2-chloro-1-(4-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isochromen-10-yl)piperazin-1-yl)ethane-1-one (compound 5)

[0064]

[0065] Compound 5 was obtained by following the synthesis method described in Example 3. The product was a yellow oily liquid with a yield of 53.7%. 1HNMR (400 MHz, CDCl3) δ 5.27 (s, 1H), 4.06 (s, 2H), 4.04 (d, J = 10.4 Hz, 1H), 3.62 (t, J = 7.3 Hz, 3H), 3.56 – 3.43 (m, 2H), 3.08 – 2.94 (m, 2H), 2.77 – 2.64 (m, 2H), 2.63 – 2.53 (m, 1H), 2.40 – 2.28 (m, 1H), 2.05 – 1.95 (m, 1H), 1.92 – 1.81 (m, 1H), 1.76 – 1.65 (m, 2H), 1.58 – 1.39 (m, 3H), HRMS (ESI) calcd for C 21 H 33 ClN2O5 [M+H] + 429.2151, found 429.2151.

[0066] Example 6: Preparation of 2,2-dichloro-1-(4-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isochromen-10-yl)piperazin-1-yl)ethane-1-one (compound 6)

[0067]

[0068] Compound 6 was obtained by following the synthesis method described in Example 3. The product was a yellow oily liquid with a yield of 45.4%. 1HNMR (400 MHz, CDCl3) δ 6.20 (s, 1H), 5.28 (s, 1H), 4.05 (d, J = 10.2 Hz, 1H), 3.68 (dd, J = 20.7, 11.0 Hz, 4H), 3.12 – 2.98 (m, 2H), 2.81 – 2.67 (m, 2H),2.65 – 2.52 (m, 1H), 2.41 – 2.28 (m, 1H), 2.05 – 1.95 (m, 1H), 1.92 – 1.81(m, 1H), 1.75 – 1.68 (m, 2H), 1.58 (s, 1H), 1.55 – 1.40 (m, 2H), HRMS (ESI) calcd for C 21 H 32 Cl2N2O5 [M+H] + 463.1762, found 463.1761.

[0069] Example 7: Preparation of (E)-4-oxo-4-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxane-heptano[4,3-i]isochroman-10-yl)piperazin-1-yl)but-2-enoic acid ethyl ester (compound 7)

[0070]

[0071] Compound 7 was obtained by following the synthesis method described in Example 3. The product was a yellow oily liquid with a yield of 87.0%. 1HNMR (400 MHz, CDCl3) δ 7.38 (d, J = 15.4 Hz, 1H), 6.74 (d, J = 15.4 Hz, 1H), 5.27 (s, 1H), 4.25 (q, J = 7.1 Hz, 2H), 4.04 (d, J = 10.2 Hz, 1H), 3.69 (s,2H), 3.62 – 3.48 (m, 2H), 3.06 – 2.96 (m, 2H), 2.74 – 2.65 (m, 2H), 2.65 –2.52 (m, 1H), 2.40 – 2.28 (m, 1H), 2.04 – 1.94 (m, 1H), 1.91 – 1.81 (m, 1H),1.75 – 1.66 (m, 2H), 1.58 – 1.39 (m, 3H), 1.38 – 1.34 (m, 4H), 1.33 – 1.29(m, 4H), 1.07 – 0.97 (m, 1H), 0.95 (d, J = 6.2 Hz, 3H), 0.82 (d, J = 7.2 Hz,3H); HRMS (ESI) calcd for C 25 H 38 N₂O₇ [M+H] + 479.2752, found 479.2750.

[0072] Example 8: Preparation of (E)-4-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxane-heptano[4,3-i]isochroman-10-yl)piperazin-1-yl)but-2-enoic acid methyl ester (compound 8)

[0073]

[0074] Compound 2 (300.0 mg, 0.851 mmol) was dissolved in dichloromethane under nitrogen protection. DIPEA (296.4 μL, 1.702 mmol) and methyl (E)-4-bromo-2-butenoate (110.0 μL, 0.936 mmol) were added, and the mixture was stirred at room temperature for 12 h. The reaction progress was monitored by TLC. The reaction solution was washed three times each with water and saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to give compound 8, with a mass of 59 mg. The product was a yellow oily liquid, with a yield of 15.4%. 1H NMR (400 MHz, CDCl3) δ 6.97 (dt, J = 15.8, 6.2 Hz, 1H), 5.99 (dt, J = 15.7, 1.6 Hz, 1H), 5.26 (s, 1H), 4.01 (d, J = 10.2 Hz, 1H), 3.73 (s, 3H), 3.21 – 3.05 (m, 2H), 3.05 – 2.96 (m, 2H), 2.70 (dt, J = 10.9, 4.7 Hz, 2H), 2.62 – 2.53 (m, 1H), 2.47 (s, 4H), 2.39 – 2.28 (m, 1H), 2.04 – 1.94 (m, 1H), 1.91 – 1.80 (m, 1H),1.72 – 1.66 (m, 2H), 1.58 – 1.39 (m, 3H), 1.37 (s, 3H), 1.36 – 1.29 (m, 2H),1.05 – 0.96 (m, 1H), 0.94 (d, J = 6.2 Hz, 3H), 0.79 (d, J = 7.2 Hz, 3H); HRMS(ESI) calcd for C 24 H 38 N₂O₆ [M+H] + 451.2803, found 451.2804.

[0075] Example 9: Preparation of 2-((4-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanene-10-yl)piperazin-1-yl)meth)methyl acrylate (compound 9)

[0076]

[0077] Compound 9 was obtained by following the synthesis method described in Example 8. The product was a yellow oily liquid with a yield of 58.3%. 1HNMR (400 MHz, CDCl3) δ 6.27 (s, 1H), 5.80 (s, 1H), 5.26 (s, 1H), 4.01 (d, J =10.2 Hz, 1H), 3.75 (s, 3H), 3.20 (s, 2H), 3.00 (t, J = 5.6 Hz, 2H), 2.72 –2.63 (m, 2H), 2.60 – 2.53 (m, 1H), 2.47 (s, 4H), 2.38 – 2.29 (m, 1H), 2.03 –1.96 (m, 1H), 1.89 – 1.81 (m, 1H), 1.72 – 1.65 (m, 2H), 1.57 – HRMS (ESI) calcd for C 24 H 38 N₂O₆ [M+H] + 451.2803, found 451.2801.

[0078] Example 10: Preparation of 2-((4-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanene-10-yl)piperazin-1-yl)meth)isopropyl acrylate (compound 10)

[0079]

[0080] Compound 10 was obtained by referring to the synthesis method of Example 8. The product was a yellow oily liquid with a yield of 44.7%. 1H NMR (400 MHz, CDCl3) δ 6.21 (d, J = 1.9 Hz, 1H), 5.74 (q, J = 1.7 Hz, 1H), 5.25 (s, 1H), 5.12 – 4.99 (m, 1H), 3.99 (d, J = 10.2 Hz, 1H), 3.24 – 3.11 (m,2H), 2.98 (dt, J = 10.8, 4.8 Hz, 2H), 2.65 (dt, J = 10.9, 4.5 Hz, 2H), 2.59 –2.51 (m, 1H), 2.51 – 2.40 (m, 4H), 2.37 – 2.27 (m, 1H), 2.02 – 1.94 (m, 1H),1.88 – 1.79 (m, 1H), 1.74 – 1.62 (m, 2H), 1.54 – 1.34 (m, 6H), 1.34 – 1.30(m, 1H), 1.27 – 1.24 (m, 7H), 1.04 – 0.95 (m, 1H), 0.93 (d, J = 6.3 Hz, 3H), 0.78 (d, J = 7.2 Hz, 3H); HRMS (ESI) calcd for C 26 H 42 N₂O₆ [M+H] + 479.3116, found 479.3120.

[0081] Example 11: Preparation of 2-(4-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isochromen-10-yl)piperazin-1-yl)ethanol (compound 11)

[0082]

[0083] Compound 11 was obtained by referring to the synthesis method of Example 2. The product was a colorless, transparent, oily liquid with a yield of 30.0%. 1H NMR (400 MHz, CDCl3) δ 5.27 (s, 1H), 4.04 (d, J = 10.2 Hz, 1H), 3.65(t, J = 5.3 Hz, 2H), 3.10 – 3.01 (m, 2H), 2.77 (d, J = 10.3 Hz, 2H), 2.69 –2.52 (m, 7H), 2.41 – 2.28 (m, 1H), 2.03 – 1.97 (m, 1H), 1.91 – 1.80 (m, 2H), 1.73 – 1.67 (m, 2H), 1.58 – 1.48 (m, 2H), 1.44 (dd, J = 13.5, 4.9 Hz, HRMS (ESI) calcd for C 21 H 36 N₂O₅ [M+H] + 397.2697, found 397.2696.

[0084] Example 12: Preparation of fumaric acid (2-(4-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isochromen-10-yl)piperazin-1-yl)ethyl)ethyl ester (compound 12)

[0085]

[0086] Using compound 11 as a starting material, compound 12 was obtained by following the synthesis method in Example 3. The product was a yellow oily liquid with a yield of 18.3%. 1H NMR (400 MHz, CDCl3) δ 6.86 (s, 2H), 5.26 (s, 1H), 4.36 (s, 2H), 4.26 (q, J = 7.1 Hz, 2H), 4.02 (d, J = 10.2 Hz, 1H), 3.02 (s, 2H), 2.71 (s, 4H), 2.61 – 2.47 (m, 4H), 2.34 (td, J = 13.9, 3.9 Hz, 1H), 2.04 –1.94 (m, 1H), 1.90 – 1.81 (m, 1H), 1.73 – 1.66 (m, 2H), 1.56 – 1.38 (m, 4H),1.36 (s, 3H), HRMS (ESI) calcd forC 27 H 42 N₂O₈ [M+H] + 523.3014, found 523.3012.

[0087] Example 13: Preparation of methyl methacrylate (compound 13) of 2-((4-((3R,5aS,6R,8aS,9R,10R,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanen-10-yl)-1,4-diazacycloheptane-1-yl)meth)

[0088]

[0089] Compound 13 was obtained by following the synthesis methods of Examples 2 and 8. The product was a yellow oily liquid with a yield of 37.1%. 1H NMR (400 MHz, CDCl3) δ 6.22 (dq, J = 3.4, 1.7 Hz, 1H), 5.78 (q, J =1.6 Hz, 1H), 5.27 (s, 1H), 4.06 (d, J = 10.2 Hz, 1H), 3.73 (s, 3H), 3.33 (t,J = 1.3 Hz, 2H), 3.18 – 3.08 (m, 2H), 2.93 – 2.78 (m, 2H), 2.77 – 2.71 (m,1H), 2.71 – 2.68 (m, 2H), 2.67 – 2.62 (m, 1H), 2.57 – 2.43 (m, 1H), 2.39 –2.26 (m, 1H), 2.02 – 1.94 (m, 1H), 1.88 – 1.80 (m, 1H), 1.79 – 1.73 (m, 2H), 1.71 – 1.65 (m, 2H), 1.54 – 1.40 (m, 3H), 1.37 (s, 3H), 1.36 – 1.30 (m, 2H),1.04 – 0.95 (m, 1H), 0.93 (d, J = 6.3 Hz, 3H), 0.82 (d, J = 7.1 Hz, 3H); HRMS(ESI) calcd for C 25 H 40 N₂O₆ [M+H] + 465.2960, found 465.2958.

[0090] Example 14: Preparation of (3R,5aS,6R,8aS,9R,10S,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanene-10-yl-piperazine-1-carboxylic acid ester (compound 14)

[0091]

[0092] DHA (1990.5 mg, 7.0 mmol) was dissolved in dichloromethane, and N,N'-carbonyldiimidazole (CDI, 1362.1 mg, 8.4 mmol) was added. The mixture was stirred at room temperature for 10 min. Triethylamine (1167.6 μL, 8.4 mmol) and piperazine (603.0 mg, 7.0 mmol) were added, and the mixture was stirred at room temperature for 12 h. The reaction progress was monitored by TLC, and the mixture was concentrated under reduced pressure and purified by column chromatography to give compound 14. The product was a yellow oily liquid in 83.6% yield. 1H NMR (400 MHz, CDCl3) δ 5.68 (d, J = 9.8 Hz, 1H), 5.42(s, 1H), 3.53 (s, 1H), 3.48 – 3.41 (m, 3H), 2.86 – 2.76 (m, 4H), 2.61 – 2.47(m, 1H), 2.40 – 2.27 (m, 1H), 2.05 – 1.95 (m, 1H), 1.91 – 1.79 (m, 1H), 1.78– 1.65 (m, 4H), 1.58 (dt, J = 13.7, 4.3 Hz, 1H), 1.52 – 1.38 (m, 5H), 1.35(dd, J = HRMS (ESI) calcd for C 20 H 32 N₂O₆ [M+H] + 397.2334, found 397.2331.

[0093] Example 15: Preparation of (3R,5aS,6R,8aS,9R,10S,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanene-10-yl 4-(2-(methoxycarbonyl)allyl)piperazine-1-carboxylic acid ester (compound 15)

[0094]

[0095] Using compound 14 as a starting material, compound 15 was obtained by following the synthesis method in Example 8. The product was a yellow oily liquid with a yield of 27.7%. 1H NMR (400 MHz, CDCl3) δ 6.25 (d, J = 1.7 Hz, 1H), 5.75 (q,J = 1.5 Hz, 1H), 5.67 (d, J = 9.8 Hz, 1H), 5.42 (s, 1H), 3.74 (s, 3H), 3.56(s, 1H), 3.53 – 3.45 (m, 3H), 3.19 (t, J = 1.1 Hz, 2H), 2.61 – 2.48 (m, 1H), 2.47 – 2.29 (m, 5H), 2.04 – 1.97 (m, 1H), 1.90 – 1.82 (m, 1H), 1.78 – 1.66(m, 2H), 1.59 (dt, J = 13.8, 4.4 Hz, 1H), 1.53 – 1.38 (m, 5H), 1.38 – 1.27(m, 2H), 1.04 – 0.96 (m, 1H), 0.94 (d, J = 6.0 Hz, 3H), 0.84 (d, J = 7.1 Hz,3H); HRMS (ESI) calcd for C 25 H 38 N₂O₈ [M+H] + 495.2701, found 495.2702.

[0096] Example 16: Preparation of (3R,5aS,6R,8aS,9R,10S,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanen-10-yl 4-(2-(ethoxycarbonyl)allyl)piperazine-1-carboxylic acid ester (compound 16)

[0097]

[0098] Using compound 14 as a starting material, compound 16 was obtained by following the synthesis method in Example 8. The product was a yellow oily liquid with a yield of 45.6%. 1H NMR (400 MHz, CDCl3) δ 6.25 (s, 1H), 5.73 (s, 1H), 5.68 (d, J = 9.7, 1H), 5.42 (d, J = 2.4 Hz, 1H), 4.20 (qd, J = 7.3, 2.3 Hz, 2H), 3.58 (d, J = 14.9 Hz, 1H), 3.53 – 3.43 (m, 3H), 3.19 (s, 2H), 2.61 – 2.48 (m,1H), 2.45 – 2.29 (m, 5H), 2.00 (dd, J = 14.2, 3.7 Hz, 1H), 1.91 – 1.81 (m,1H), 1.80 – 1.64 (m, 2H), 1.59 (dt, J = 14.4, 4.1 Hz, 1H), 1.53 – 1.43 (m,1H), 1.41 (s, 3H), 1.39 – 1.32 (m, 1H), 1.32 – 1.25 (m, 5H), 1.05 – 0.96 (m,1H), 0.94 (d, J = 6.0 Hz, 3H), 0.84 (d, J = 7.3 Hz, 3H); HRMS (ESI) calcd forC 26 H 40 N₂O₈ [M+H] + 509.2858, found 509.2855.

[0099] Example 17: Preparation of (3R,5aS,6R,8aS,9R,10S,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanene-10-yl 4-(2-(isopropoxycarbonyl)allyl)piperazine-1-carboxylic acid ester (compound 17)

[0100]

[0101] Using compound 14 as a starting material, compound 17 was obtained by following the synthesis method in Example 8. The product was a yellow oily liquid with a yield of 30.9%. 1H NMR (400 MHz, CDCl3) δ 6.21 (s, 1H), 5.72 – 5.64 (m, 2H), 5.43 (s, 1H), 5.12 – 5.01 (m, 1H), 3.58 (d, J = 15.0 Hz, 1H), 3.47 (s, 3H),3.19 (s, 2H), 2.60 – 2.48 (m, 1H), 2.47 – 2.29 (m, 5H), 2.05 – 1.96 (m, 1H),1.90 – 1.83 (m, 1H), 1.79 – 1.64 (m, 2H), 1.59 (dt, J = 14.2, 4.0 Hz, 1H),1.53 – 1.43 (m, 1H), 1.41 (s, 3H), 1.38 – 1.28 (m, 2H), 1.27 – 1.24 (m, 7H), 1.05 – 0.96 (m, 1H), 0.94 (d, J = 6.0 Hz, 3H), 0.84 (d, J = 7.1 Hz, 3H); HRMS(ESI) calcd for C 27 H 42 N₂O₈ [M+H] + 523.3014, found 523.3012.

[0102] Example 18: Preparation of (3R,5aS,6R,8aS,9R,10S,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanen-10-yl-3-((2-(methoxycarbonyl)allyl)oxy)azacyclobutane-1-carboxylic acid ester (compound 18)

[0103]

[0104] DHA (1990.5 mg, 7.0 mmol) and CDI (1362.1 mg, 8.4 mmol) were dissolved in dichloromethane and stirred at room temperature for 10 min. Triethylamine (1167.6 μL, 8.4 mmol) and 3-hydroxyazacyclobutane hydrochloride (766.9 mg, 7.0 mmol) were added, and the mixture was stirred at room temperature for 12 h. The reaction progress was monitored by TLC, and the mixture was concentrated under reduced pressure and purified by column chromatography to obtain crude (3R,5aS,6R,8aS,9R,10S,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxy[1,2]dioxane-heptano[4,3-i]isocyanen-10-yl-3-hydroxyazacyclobutane-1-carboxylate. The product was a yellow oily liquid with a yield of 63.8%, and was defined as intermediate Z1.

[0105] Intermediate Z1 (200.0 mg, 0.522 mmol) was dissolved in dichloromethane and kept in the dark. Silver oxide (133.0 mg, 0.574 mmol) and methyl 2-(bromomethyl)acrylate (69 μL, 0.574 mmol) were added, and the mixture was stirred at room temperature for 12 h. The reaction progress was monitored by TLC. The silver oxide solid was filtered through diatomaceous earth, and the diatomaceous earth was washed with dichloromethane. The filtrate was concentrated under reduced pressure and purified by column chromatography to obtain compound 18. The product was a yellow oily liquid with a yield of 90.0%. 1H NMR (400 MHz, CDCl3) δ 6.30(q, J = 1.3 Hz, 1H), 5.87 (q, J = 1.6 Hz, 1H), 5.62 (d, J = 9.8 Hz, 1H), 5.41(s, 1H), 4.34 – 4.29 (m, 1H), 4.18 – 4.04 (m, 4H), 3.92 (d, J = 3.7 Hz, 1H), 3.89 (d, J = 4.8 Hz, 1H), 3.75 (s, 3H), 2.57 – 2.43 (m, 1H), 2.38 – 2.28 (m,1H), 2.05 – 1.95 (m, 1H), 1.89 – 1.85 (m, 1H), 1.85 – 1.81 (m, 1H), 1.77 –1.65 (m, 2H), 1.57 (dt, J = 13.8, 4.4 Hz, 1H), 1.51 – 1.41 (m, 1H), 1.41 –1.39 (m, 3H), 1.39 – 1.28 (m, 2H), 1.03 – 0.95 (m, 1H), 0.93 (d, J = 6.1 Hz, 3H), 0.82 (d, J = 7.1 Hz, 3H); HRMS (ESI) calcd for C 24 H 35 NO9 [M+H] + 482.2385, found 482.2382.

[0106] Example 19: Preparation of (3R,5aS,6R,8aS,9R,10S,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanene-10-yl-3-((2-(ethoxycarbonyl)allyl)oxy)azacyclobutane-1-carboxylic acid ester (compound 19)

[0107]

[0108] Using intermediate Z1 as a starting material, compound 19 was obtained by following the synthesis method described in Example 18. The product was a yellow oily liquid with a yield of 67.4%. 1H NMR (400 MHz, CDCl3) δ 6.31 (q, J = 1.3 Hz, 1H), 5.87 (q,J = 1.6 Hz, 1H), 5.63 (d, J = 9.8 Hz, 1H), 5.42 (s, 1H), 4.35 – 4.31 (m, 1H), 4.22 (q, J = 7.1 Hz, 2H), 4.18 – 4.12 (m, 2H), 4.11 (s, 2H), 3.94 (d, J = 5.0Hz, 1H), 3.91 (d, J = 4.8 Hz, 1H), 2.58 – 2.45 (m, 1H), 2.41 – 2.29 (m, 1H),2.06 – 1.96 (m, 1H), 1.92 – 1.81 (m, 1H), 1.80 – 1.65 (m, 3H), 1.59 (dt, J =13.9, 4.4 Hz, 1H), 1.52 – 1.44 (m, 1H), 1.42 (s, 3H), 1.41 – 1.32 (m, 2H),1.30 (t, 3H), 1.05 – 0.96 (m, 1H), 0.94 (d, J = 6.1 Hz, 3H), 0.84 (d, J = 7.1Hz, 3H); HRMS (ESI) calcd for C 25 H 37 NO9 [M+H] + 496.2542, found 496.2544.

[0109] Example 20: Preparation of (3R,5aS,6R,8aS,9R,10S,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanen-10-yl-3-((2-(isopropoxycarbonyl)allyl)oxy)azacyclobutane-1-carboxylic acid ester (compound 20)

[0110]

[0111] Using intermediate Z1 as a starting material, compound 20 was obtained by following the synthesis method of Example 18. The product was a yellow oily liquid with a yield of 36.2%. 1H NMR (400 MHz, CDCl3) δ 6.28 (q, J = 1.3 Hz, 1H), 5.84 (q,J = 1.7 Hz, 1H), 5.63 (d, J = 9.8 Hz, 1H), 5.42 (s, 1H), 5.15 – 5.01 (m, 1H),4.35 – 4.30 (m, 1H), 4.19 – 4.06 (m, 4H), 3.93 (d, J = 4.9 Hz, 1H), 3.91 (d,J = 3.5 Hz, 1H), 2.58 – 2.45 (m, 1H), 2.40 – 2.30 (m,1H), 2.05 – 1.97 (m,1H), 1.92 – 1.81 (m, 1H), 1.77 – 1.66 (m, 2H), 1.58 (dt, J = 13.8, 4.4 Hz,1H), 1.52 – 1.43 (m, 1H), 1.41 (s, 3H), 1.40 – 1.30 (m, 2H), 1.27 HRMS (ESI) calcd for C 26 H 39 NO9 [M+Na] + 532.2518, found 532.2520.

[0112] Example 21: Preparation of fumaric acid ((3R,5aS,6R,8aS,9R,10S,12R,12aR)-3,6,9-trimethyldecahydro-12H-3,12-epoxydioxyheptano[4,3-i]isocyanen-10-yl)oxy)carbonyl)azacyclobutane-3-yl)ethyl ester (compound 38)

[0113]

[0114] Using intermediate Z1 as a raw material, compound 21 was obtained by following the synthesis method in Example 3. The product was a yellow oily liquid with a yield of 22.4%. 1H NMR (400 MHz, CDCl3) δ 6.95 – 6.81 (m, 2H), 5.65 (d, J =9.8 Hz, 1H), 5.44 (s, 1H), 5.33 – 5.23 (m, 1H), 4.41 – 4.32 (m, 1H), 4.27 (q,J = 7.1 Hz, 2H), 4.06 – 3.98 (m, 1H), 2.60 – 2.47 (m, 1H), 2.43 – 2.30 (m,1H), 2.06 – 2.00 (m, 1H), 1.96 – 1.83 (m, 1H), 1.82 – 1.66 (m, 2H), 1.64 –1.57 (m, 4H), 1.54 – 1.45 (m, 1H), 1.43 (s, 3H), 1.42 – 1.36 (m, 1H), 1.35 –1.30 (m, 4H), 1.06 – 0.98 (m, 1H), 0.96 (d, J = 6.0 Hz, 3H), 0.85 (d, J = 7.1Hz, 3H); HRMS (ESI) calcd for C 25 H 35 NO 10 [M+Na] + 532.2154, found 532.2149.

[0115] Example 22: In vitro test of the inhibitory effect of artemisinin derivatives on NLRP3 inflammasome.

[0116] THP-1 cells (human monocytic leukemia cells) were aliquoted into 96-well plates with RPMI-1640 medium containing PMA (phorbol ester, 0.1 μg / mL), 1% penicillin antibiotics, and 10% serum, at a density of 8 × 10⁸ cells per well. 5 Cells were cultured for differentiation in an incubator for 24 hours. The supernatant was discarded, and 100 μL of RPMI-1640 medium containing bacterial lipopolysaccharide (LPS, 1 μg / mL), 1% antibiotics, and 10% serum was added to each well for 3 hours. Then, different concentrations of artemisinin derivatives were added for 1 hour, followed by nigericin (10 μM) for 1 hour. The cell supernatant was then collected, and the absorbance values ​​were measured using a Human IL-1β ELISA kit to calculate the inhibitory effect of the compound of this invention on the NLRP3 inflammasome.

[0117] The results are shown in Table 1: Most of the artemisinin derivatives prepared in this invention have a strong inhibitory effect on the NLRP3 inflammasome.

[0118] Table 1. Effects of artemisinin derivatives on NLRP3 inflammasomes in THP-1 cells.

[0119] Inhibitory activity of activation-mediated IL-1β release (IC50) 50 (μM)

[0120]

[0121] Example 23: Effects of compounds 9, 16 and 21 on the viability of THP-1 and J774A.1 cells

[0122] (1) THP-1 cells were aliquoted into 96-well plates with RPMI-1640 medium containing PMA (0.1 μg / mL), 1% penicillin antibiotics, and 10% serum, with 4 × 10⁶ cells per well. 5 Cells were cultured for differentiation in an incubator for 24 hours. The supernatant was discarded, and different concentrations of artemisinin derivatives were added to each well for 2 hours, followed by CCK-8 reagent for another 2 hours. Absorbance values ​​were measured using a microplate reader, and cell viability was calculated for each group based on the control group. Figure 1 AC in the middle.

[0123] (2) J774A.1 cells (mouse mononuclear macrophages) were aliquoted into 96-well plates using DMEM medium containing 10% serum, with 5 × 10⁶ cells per well. 5 Cells were incubated overnight in an incubator. The supernatant was discarded, and different concentrations of artemisinin derivatives were added to each well for 2 hours. Cell viability was then measured for each group according to the method described in Example (1). Figure 1 (DF in the middle).

[0124] The results showed that compounds 9, 16, and 21 did not reduce the cell viability of THP-1 cells after 2 hours of treatment, while compound 21 could enhance cell viability at high concentrations (30 μM and 40 μM). When the three compounds were treated with J774A.1 cells for 2 hours, compounds 9 and 16 showed some cytotoxicity at high concentrations, while compound 21 showed no significant toxicity and even enhanced cell viability at a high concentration (40 μM). This indicates that compound 21 can effectively inhibit inflammation and has low toxicity.

[0125] Example 24: Compound 21 specifically inhibits the activation of NLRP3 inflammasome in J774A.1 cells in vitro.

[0126] 1. NLRP3 inflammasome activation and IL-1β detection: J774A.1 cells were divided into 96-well plates, 5 × 10⁶ cells per well. 5Cells were seeded overnight. The supernatant was discarded, and 100 μL of DMEM medium containing 10% serum and bacterial lipopolysaccharide (LPS, 1 μg / mL) was added to each well for 5 hours. Then, different concentrations (0.125 μM, 0.25 μM, 0.5 μM, 1 μM, 2 μM, 4 μM) of compound 21 were added for 1 hour, followed by treatment with nigericin (100 μM) for 1 hour. The cell supernatant was collected, and the absorbance was measured using a Mouse IL-1β ELISA kit.

[0127] The results are as follows Figure 2 As shown, in the J774A.1 cell model with NLRP3 inflammasome activation, compound 21 can inhibit IL-1β secretion in a concentration-dependent manner. The IC50 of the inhibitory effect of compound 21 on the NLRP3 inflammasome was calculated. 50 The value is 1.2 ± 0.1 μM.

[0128] 2. Western blot analysis of proteins: J774A.1 cells were divided into 6-well plates, 5 × 10⁶ cells per well. 6 Cells were seeded overnight. The supernatant was discarded, and each well was treated with 2 mL of DMEM medium containing 10% serum and bacterial lipopolysaccharide (LPS, 1 μg / mL) for 5 hours. Then, different concentrations (1 μM, 3 μM, 9 μM) of compound 21 were added for 1 hour, followed by treatment with Nigericin (100 μM) for 1 hour. The supernatant was extracted and preserved. J774A.1 cell samples were lysed in RIPA lysis buffer with protease inhibitors at 4°C for 10 minutes. Proteins in the lysate or supernatant were separated using a 10% SDS-polyacrylamide gel electrophoresis, transferred to a PVDF membrane, and analyzed by Western blotting with anti-mouse IL-1β antibody, anti-ASC antibody, anti-casepase-1 antibody, anti-NLRP3 antibody, and anti-β-actin antibody.

[0129] The results are as follows Figure 3 As shown, Western blot experiments revealed that compound 21 in the supernatant inhibited caspase-1 (p20) maturation and IL-1β secretion in a dose-dependent manner. In cell lysate, compound 21 inhibited the expression of pro-IL-1β and NLRP3 in a concentration-dependent manner, but did not affect pro-caspase-1 and ASC; compound 21 also inhibited the cleavage of GSDMD.

[0130] Example 25: Compound 21 inhibits the release of tumor necrosis factor-α mediated by lipopolysaccharide.

[0131] (1) THP-1 cells were aliquoted into 96-well plates with RPMI-1640 medium containing PMA (0.1 μg / mL), 1% penicillin antibiotics, and 10% serum, with 4 × 10⁶ cells per well. 5 Cells were cultured for differentiation in an incubator for 24 hours. The supernatant was discarded, and different concentrations of artemisinin derivatives were added to each well for 2 hours, followed by incubation in serum-depleted medium (Opti-MEM™ I) containing lipopolysaccharide (0.5 μg / mL) for 24 hours. Absorbance values ​​were measured using a Human TNF-α ELISA kit.

[0132] The results are as follows Figure 4 As shown in A, the IC50 of compound 21 of the present invention in inhibiting LPS-mediated TNF-α release in THP-1 cells was calculated. 50 The value is 10.3 ± 0.1 μM.

[0133] (3) Divide J774A.1 cells into 96-well plates, 5 × 10⁶ cells per well. 5 Cells were seeded overnight. The supernatant was discarded, and different concentrations of artemisinin derivatives were added to each well for 2 hours, followed by incubation in serum-depleted medium (Opti-MEM™ I) containing lipopolysaccharide (0.5 μg / mL) for 24 hours. The absorbance values ​​were measured using a Mouse TNF-α ELISA kit.

[0134] The results are as follows Figure 4 As shown in B, the IC50 of compound 21 of the present invention in inhibiting LPS-mediated TNF-α release in J774A.1 cells was calculated. 50 The value is 8.5 ± 1.6 μM.

[0135] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A dihydroartemisinin derivative of formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, in, W is selected from: , , or ; R is selected from: or ; R 1 and R 2 Each and every one is independently selected from: C1-C6 alkyl groups; n1 and n2 are independently selected from 0, 1, 2, 3 or 4, and n1 + n2 = 0~4; m1 and m2 are each independently selected from: 1, 2, 3, 4 or 5, and m1 + m2 = 2~6; y is selected from: 0, 1, 2 or 3.

2. The dihydroartemisinin derivative or its pharmaceutically acceptable salt or its stereoisomer according to claim 1, characterized in that, n1 + n2 = 1, 2, or 3; And / or, m1+m2=2, 3 or 4.

3. The dihydroartemisinin derivative or its pharmaceutically acceptable salt or its stereoisomer according to claim 1, characterized in that, W is selected from: , , , or .

4. The dihydroartemisinin derivative or its pharmaceutically acceptable salt or its stereoisomer according to claim 1, characterized in that, R is selected from: or .

5. The dihydroartemisinin derivative or its pharmaceutically acceptable salt or its stereoisomer according to any one of claims 1-4, characterized in that, R 1 and R 2 Each of the following is independently selected from: methyl, ethyl, n-propyl, and isopropyl.

6. The dihydroartemisinin derivative or its pharmaceutically acceptable salt or its stereoisomer according to any one of claims 1-3, characterized in that, R is selected from: 。 7. The dihydroartemisinin derivative or its pharmaceutically acceptable salt or its stereoisomer according to claim 1, characterized in that, The dihydroartemisinin derivative is selected from the following compounds: 。 8. The use of the dihydroartemisinin derivative or a pharmaceutically acceptable salt thereof or a stereoisomer thereof as described in any one of claims 1-7 in the preparation of an NLRP3 inflammasome inhibitor.

9. The use of the dihydroartemisinin derivative of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of a medicament for the prevention and / or treatment of diseases associated with NLRP3 inflammasome activation.

10. A pharmaceutical composition for the prevention and / or treatment of diseases associated with NLRP3 inflammasome activation, characterized in that, It is prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient includes a dihydroartemisinin derivative as described in any one of claims 1-7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

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