A sesquiterpene compound extracted and isolated from Ainsliaea axilliflora and its application

Abstract

The present invention discloses a sesquiterpenoid compound extracted and separated from the hyacinthus annuus. The sesquiterpenoid compound is at least one of compound 1 and compound 2. The molecular formula of compound 1 is C 20 H 24 O4, the structural formula is shown in formula 1; the molecular formula of compound 2 is C 20 H 24 O5, the structural formula is shown in Formula 2. The present invention also discloses the use of the above-mentioned sesquiterpenoid compounds extracted and separated from the axillary wormwood. Compounds 1 and 2 have the effects of inhibiting inflammasome activation and blocking cell pyroptosis, and can be used to prepare products that inhibit inflammasome activation and / or block cell pyroptosis, and can also be used to prepare products for preventing or treating NLRP3 inflammasome-related diseases.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a sesquiterpenoid compound extracted and separated from Lepidoptera: ... Background Art

[0002] Pyroptosis is a new type of programmed cell death, first proposed by Professor Cookson of the University of Washington in 2001. Pyroptosis is a phenomenon in which numerous pores form in the cell membranes of various cells (such as macrophages) over a short period of time. Pyroptosis disrupts the osmotic pressure balance inside and outside the cell, increasing the osmotic pressure within the cell, leading to cell swelling and rupture, and the release of a large amount of internal cellular substances.

[0003] Existing research results have confirmed that inflammasome activation leads to the activation of cysteine-containing aspartate proteinase-1 (Caspase-1), thereby triggering cell pyroptosis and a strong inflammatory response.

[0004] The activated Caspase-1 protease will continue to cleave a specific site in the middle of the gastrosin D (GSDMD) protein, leading to the formation of the N-terminal fragment of GSDMD (GSDMD-NT). The protein will then localize to the cell membrane and punch holes in the cell membrane, leading to cell pyroptosis and the release of a large amount of internal cell substances.

[0005] At the same time, the activated Caspase-1 protease will induce the maturation of pro-inflammatory factors such as IL-1β. A large number of mature pro-inflammatory factors are released outward through the holes in the cell membrane, leading to a strong inflammatory response.

[0006] Hallmarks of inflammasome activation are the cleavage of caspase-1 and the maturation of IL-1β, both of which are released extracellularly and can be detected by immunoblotting (WB) and enzyme-linked immunosorbent assay (ELISA). A hallmark of pyroptosis is the breakdown of cell membrane integrity, which can be detected by intracellular propidium iodide (PI) staining, extracellular lactine dehydrogenase (LDH) release, and GSDMD-NT formation (WB).

[0007] Inflammasome activation-induced pyroptosis is associated with a variety of inflammatory diseases and inflammatory aging, including rheumatoid arthritis, gouty arthritis, chemical liver damage, silicosis, UV sunburn, acne caused by Propionibacterium acnes, and dandruff caused by Malassezia. Therefore, inhibiting pyroptosis and blocking the inflammatory cytokine storm can improve a variety of diseases and alleviate aging.

[0008] However, the existing therapeutic drugs used for diseases have "pan-cytotoxicity" or systemic toxicity. For example, methotrexate, colchicine, glucocorticoids, etc. all have certain toxicity. Although these drugs can control inflammatory responses by inhibiting the proliferation of all cells in the human body and indiscriminately blocking a wide range of intracellular signaling pathways, while suppressing inflammatory immune cells, they also damage normal immune cells and other body cells (such as germ cells). They are passive and broad-based interventions that cannot be targeted and accurately inhibit the maturation of inflammatory factors such as IL-1β from the source to block cell pyroptosis.

[0009] Inflammasome inhibitors can precisely inhibit the maturation of inflammatory factors such as IL-1β at the source, blocking pyroptosis. Because inflammasomes are not activated in normal cells, these inhibitors have minimal impact on normal immune cells and other body cells, making them an effective strategy for precise disease intervention, a form of proactive precision intervention.

[0010] Substances disclosed in the prior art that can be used as inflammasome inhibitors include: piperlongamide (CN202010794165.6 "Application of piperlongamide in inhibiting NLRP3 inflammasome activation"), capsaicin (202110196432.4 "Application of capsaicin in inhibiting NLRP3 inflammasome activation"), ipriflavone (CN202210035588.9 "Application of ipriflavone in inhibiting NLRP3 inflammasome activation") There are many types of inflammasome inhibitors, such as wedeliolide (CN202211119761.X "Application of wedeliolide in the preparation of inflammasome activation inhibitors or drugs for treating gouty arthritis"), wedeliolide (CN202211119761.X "Application of wedeliolide in the preparation of inflammasome activation inhibitors or drugs for treating gouty arthritis"), etc. However, the performance of the above-mentioned inflammasome inhibitors varies, and the effects of the natural products currently used as inhibitors are poor.

[0011] The plant, Echeveria axillaris, is a perennial herb, reaching 50-120 cm in height and flowering from March to June and September to October. It grows primarily in valleys, along streams, or in moist forests at altitudes of 1,500-2,500 meters. It is primarily found in Yunnan, Guizhou, and Sichuan, and has also been found in India. Echeveria axillaris has hemostatic and analgesic properties and is commonly used in folk treatments for ankylosing spondylitis, traumatic bleeding, arthritis, and stomachache.

[0012] However, existing research results have not found that sesquiterpenoid compounds derived from the plant Ainsliaea pertyoides Franch. have the effect of inhibiting inflammasome activation, nor have they clarified which structure of sesquiterpenoid compounds has better inflammasome activation inhibition function. Summary of the Invention

[0013] The purpose of the present invention is to overcome the defects of the prior art and provide a sesquiterpenoid compound extracted and separated from the axillary terpenoid, which can be used as an inflammasome inhibitor for inhibiting inflammasome activation. The sesquiterpenoid compound shows an effective inhibitory effect on inflammasomes and cell pyroptosis, and can be used in the preparation of inflammasome inhibitors, blocking cell pyroptosis, and in the preparation of medicines, health foods, cosmetics, and sanitary disinfection products for preventing, treating, or improving cell pyroptosis-related diseases.

[0014] Another object of the present invention is to provide an application of sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae.

[0015] A technical solution to achieve the above purpose is: a sesquiterpenoid compound extracted and separated from the axillary wormwood, wherein the sesquiterpenoid compound is at least one of compound 1 and compound 2, and the molecular formula of compound 1 is C 20 H 24 O4, the structural formula is shown in formula (1):

[0016]

[0017] The molecular formula of compound 2 is C 20 H 24 O5, the structural formula is shown in formula (2):

[0018]

[0019] The above-mentioned sesquiterpenoid compound extracted and separated from the scutellaria baicalensis, wherein the steps of extracting and separating the sesquiterpenoid compound from the scutellaria baicalensis are as follows:

[0020] (1) The dried whole herb of Ainsliaea pertyoides Franch. was extracted with 85% methanol, and then concentrated and dried under reduced pressure to obtain an 85% methanol extract;

[0021] (2) dissolving the 85% methanol extract obtained in step (1) in water, extracting with ethyl acetate which is immiscible with water, and recovering the solvent under reduced pressure to obtain an extract;

[0022] (3) The extract obtained in step (2) was separated by D-101 macroporous resin column chromatography using a methanol / water (50:50-100:0, v / v) gradient elution to obtain five fractions (Fr.1-Fr.5);

[0023] (4) Fr.2 obtained in step (3) above was separated by silica gel column chromatography, and eluted with a gradient of chloroform / methanol (10:1-0:1, v / v) to obtain 9 fractions (Fr.2.1-Fr.2.9); Fr.3 was separated by silica gel column chromatography, and eluted with a gradient of petroleum ether / ethyl acetate (30:1-1:1, v / v) to obtain 6 fractions (Fr.3.1-Fr.3.6);

[0024] (5) Fr.2.8 and Fr.3.3 obtained in the above step (4) were separated by semi-preparative HPLC chromatography, eluted with 70% and 60% acetonitrile, respectively, to obtain compounds 1 and 2.

[0025] The present invention also provides an application of the above-mentioned sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae, wherein the sesquiterpenoid compounds are used in the preparation of products that inhibit inflammasome activation and / or block cell pyroptosis.

[0026] The above-mentioned application of sesquiterpenoid compounds extracted and separated from Lepidoptera: wherein the inflammasome is the NLRP3 inflammasome.

[0027] The above-mentioned application of sesquiterpenoid compounds extracted and separated from Coleus axillaryum, wherein the inhibition of inflammasome activation is at least one of inhibiting the cleavage of Caspase-1 protein in inflammasome-inducing cells, inhibiting the maturation of IL-1β in inflammasome-inducing cells, and inhibiting the secretion of IL-1β in inflammasome-inducing cells.

[0028] The application of the sesquiterpenoid compound extracted and separated from the axillary wormwood, wherein the IC of the sesquiterpenoid compound inhibits the secretion of IL-1β in inflammasome-induced cells 50 All were less than 10μM.

[0029] The above-mentioned application of sesquiterpenoid compounds extracted and separated from Thunbergia axillariae, wherein the blocking of cell pyroptosis is at least one of reducing the propidium iodide positivity rate of inflammasome-induced cells, inhibiting the release of lactate dehydrogenase in inflammasome-induced cells, and inhibiting the formation of GSDMD-NT in inflammasome-induced cells.

[0030] The application of the sesquiterpenoid compounds extracted and separated from the axillary terpenoids, wherein when the concentration of the sesquiterpenoid compounds is 10 μM, the propidium iodide (PI) positive inhibition rate is 15-19%, and the lactate dehydrogenase (LDH) release rate is 7-13%.

[0031] The present invention also provides an application of the above-mentioned sesquiterpenoid compound extracted and separated from Lepidoptera: wherein the sesquiterpenoid compound is used in the preparation of a product for preventing or treating NLRP3 inflammasome-related diseases.

[0032] The above-mentioned application of sesquiterpenoid compounds extracted and separated from Axillary Thunbergia serrata, wherein the NLRP3 inflammasome-related disease is at least one of gout, arthritis, alcoholic liver damage, non-alcoholic liver damage, atherosclerosis, type II diabetes, sepsis, multiple sclerosis, systemic lupus erythematosus, Alzheimer's disease, Parkinson's disease, silicosis, cold pyrin syndrome, acne, psoriasis, dandruff, tinea versicolor, seborrheic dermatitis, UV-induced skin damage, UV-induced skin tanning, UV-induced skin aging and mosquito bites.

[0033] The above-mentioned application of sesquiterpenoid compounds extracted and separated from Achyranthes axillary, wherein the dosage form of the product is at least one of tablets, dispersible tablets, lozenges, orally disintegrating tablets, sustained-release tablets, capsules, soft capsules, pills, granules, injections, powder injections or aerosol powders, films, plasters, patches, emulsions, liniments, creams and lotions.

[0034] The above-mentioned applications of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae, wherein the products include medicines, health foods, cosmetics and sanitary disinfectants.

[0035] The above-mentioned application of sesquiterpenoid compounds extracted and separated from the hyssopus axillary, wherein the cosmetics are any one of shampoo, shampoo cream, cooling oil, wind oil essence, toner, floral water, facial cream, facial mask, facial gel, sunscreen milk, sunscreen cream, sunscreen gel and sunscreen lotion.

[0036] The technical solution of extracting and separating sesquiterpenoid compounds from Achyranthes axillary and its application has the following beneficial effects:

[0037] (1) Compound 1 represented by formula (1) and Compound 2 represented by formula (2) were extracted and separated from Thunbergia axillaris for the first time; the extraction and separation method was simple and rapid, and the compounds extracted and separated from Thunbergia axillaris had a high purity of up to 95%;

[0038] (2) The sesquiterpenoid compounds of the present invention can be used to prepare products that inhibit inflammasome activation and / or block cell pyroptosis. Compound 1 and Compound 2 have the effect of inhibiting inflammasome activation and / or blocking cell pyroptosis. Compound 1 and Compound 2, at a concentration of 10 μM, can inhibit the extracellular release of LDH, inhibit the formation of GSDMD-NT, inhibit the cleavage of Caspase-1, and inhibit the maturation of IL-1β, that is, by inhibiting the activation of inflammasomes, thereby blocking the occurrence of cell pyroptosis. At a concentration of 10 μM, the inhibition rate of compound 1 and compound 2 on cell pyroptosis (LDH method, PI method) exceeded 50%, and the IC value for inhibiting IL-1β secretion (ELISA method) was 2.5%. 50 The results were all less than 10 μM, indicating that compounds 1 and 2 have the effects of inhibiting inflammasome activation and blocking cell pyroptosis; the sesquiterpenoid compounds of the present invention can be used to prepare products for preventing or treating NLRP3 inflammasome-related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 For compound 1 1 H-NMR spectrum;

[0040] Figure 2 For compound 1 13 C-NMR spectrum;

[0041] Figure 3 For compound 2 1 H-NMR spectrum;

[0042] Figure 4 For compound 2 13 C-NMR spectrum;

[0043] Figure 5 is the HSQC spectrum of compound 2;

[0044] Figure 6 is the HMBC spectrum of compound 2;

[0045] Figure 7 For compound 2 1 H- 1 H COSY spectrum;

[0046] Figure 8 is the ROESY spectrum of compound 2;

[0047] Figure 9 The key to compound 2 1 H- 1 H COSY and HMBC Related;

[0048] Figure 10The key ROESY of compound 2 ( α-face β-face) related signals;

[0049] Figure 11 are the calculated and measured ECD spectra of compound 2;

[0050] Figure 12 This is a bar graph showing the PI staining and positive rate results obtained in Example 1 (the percentage of cell pyroptosis induced by activated inflammasomes in the treatment with compound 1 indicates that the compound can effectively block cell pyroptosis);

[0051] Figure 13 This is a bar graph showing the PI staining and positive rate results obtained in Example 1 (the percentage of cell pyroptosis induced by activated inflammasomes in the treatment with compound 2 indicates that the compound can effectively block cell pyroptosis);

[0052] Figure 14 This is a bar graph of the lactate dehydrogenase (LDH) release rate of compounds 1-2 in Example 2 (proving that compounds 1-2 can effectively inhibit cell pyroptosis);

[0053] Figure 15 This is the immunoblotting result of GSDMD-NT after treatment with compound 1 in Example 3;

[0054] Figure 16 This is an immunoblot image of the experiment in Example 4 in which compound 1 inhibits Caspase-1 and IL-1β protein cleavage and blocks inflammasome activation;

[0055] Figure 17 This is an immunoblot image of the experiment in Example 4 in which compound 2 inhibited Caspase-1 and IL-1β protein cleavage and blocked inflammasome activation;

[0056] Figure 18 This is a bar graph showing the experimental results of compound 1 inhibiting the secretion of IL-1β and blocking the activation of inflammasomes in Example 5;

[0057] Figure 19 This is the immunoblotting result of compound 1 inhibiting the formation of ASC oligomers (ASC Oligomerization) in Example 6;

[0058] Figure 20 Compound 1 in Example 7 inhibits MSU-induced mitochondrial damage in J774A.1 cells;

[0059] Figure 21 Compound 1 in Example 8 inhibits the production of mitochondrial reactive oxygen species (mt-SOX) in J774A.1 cells induced by MSU;

[0060] Figure 22 Representative photos of compound 1 in Example 9 inhibiting joint swelling in mice induced by gouty arthritis induced by sodium urate crystals (MSU);

[0061] Figure 23 This is a bar graph showing the inhibition of compound 1 in Example 9 on the diameter of joint swelling in mice with gouty arthritis induced by sodium urate crystals (MSU);

[0062] Figure 24 Western blot analysis of NLRP3 inflammasome-related proteins for the inhibition of ankle arthritis in mice by compound 1 in Example 9;

[0063] Figure 25 This is a bar graph showing the results of compound 1 in Example 9 inhibiting the expression level of IL-1β in the culture supernatant of mouse ankle joints. DETAILED DESCRIPTION

[0064] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Generally, the components of the embodiments of the present invention described and shown in the drawings herein can be arranged and designed in various different configurations.

[0065] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the invention as claimed, but rather merely represents selected embodiments of the present invention. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without creative effort are also within the scope of protection of the present invention.

[0066] The technical means that are not described in detail in this application and are not used to solve the technical problems of this application are all set according to the common knowledge in this field, and can be implemented in a variety of common knowledge settings.

[0067] Unless otherwise specified, the materials and instruments used in the following examples were obtained from commercial channels.

[0068] The sesquiterpenoid compound extracted and separated from the axillary wormwood is at least one of compound 1 and compound 2. The English name of compound 1 is: 2-Butenoic acid, 3-methyl-, (3aS, 6aR, 8S, 9aR, 9bS)-dodecahydro-3, 6, 9-tris (methylene) -2-oxoazuleno [4, 5-b] furan-8-yl ester. The molecular formula of compound 1 is C 20 H24 O4, the structural formula is shown in formula (1):

[0069]

[0070] The English name of compound 2 is Ainslide C, and the molecular formula of compound 2 is C 20 H 24 O5, the structural formula is shown in formula (2):

[0071]

[0072] The preparation methods of compound 1 and compound 2 are as follows:

[0073] (1) The dried whole herb of Ainsliaea pertyoides Franch. was extracted with 85% methanol, and then concentrated and dried under reduced pressure to obtain an 85% methanol extract;

[0074] (2) dissolving the 85% methanol extract obtained in step (1) in water, extracting with ethyl acetate which is immiscible with water, and recovering the solvent under reduced pressure to obtain an extract;

[0075] (3) The extract obtained in step (2) was separated by D-101 macroporous resin column chromatography using a methanol / water (50:50-100:0, v / v) gradient elution to obtain five fractions (Fr.1-Fr.5);

[0076] (4) Fr.2 obtained in step (3) above was separated by silica gel column chromatography, and eluted with a gradient of chloroform / methanol (10:1-0:1, v / v) to obtain 9 fractions (Fr.2.1-Fr.2.9); Fr.3 was separated by silica gel column chromatography, and eluted with a gradient of petroleum ether / ethyl acetate (30:1-1:1, v / v) to obtain 6 fractions (Fr.3.1-Fr.3.6);

[0077] (5) Fr.2.8 and Fr.3.3 obtained in the above step (4) were separated by semi-preparative HPLC chromatography, eluted with 70% and 60% acetonitrile, respectively, to obtain compounds 1 and 2.

[0078] Structural identification of compound 1:

[0079] See also Figure 1 and Figure 2 , Compound 1 is a colorless oil, 1 H-NMR (400 MHz, CDCl3), δ H:6.22(1H,d,J=3.5Hz,H-13),5.72(1H,s,H-2′),5.58(1H,dd,H-3),5.50(1H,d,J=3.0Hz,H-13),5.46 (1H,t,J=2.0Hz,H-15),5.29(1H,t,J=2.0Hz,H-15),4.95(2H,d,J=9.1Hz,H-14),4.07(1H,t,J=9.3Hz, H-6),2.95(1H,m,H-1),2.87(1H,m,H-7),2.84(1H,m,H-5),2.49(1H,m,H-2),2.44(1H,m,H-9),2.25(1 H,m,H-8),2.18(3H,s,H-4′),2.16(1H,m,H-2),1.91(3H,s,H-5′),1.82(1H,m,H-9),1.46(1H,m,H-8); 13 C-NMR (100 MHz, CDCl3), δ C The molecular weight of the compound 1 was 44.8 (C-1), 34.8 (C-2), 73.9 (C-3), 148.0 (C-4), 50.4 (C-5), 84.1 (C-6), 45.4 (C-7), 30.8 (C-8), 36.9 (C-9), 148.6 (C-10), 139.8 (C-11), 170.1 (C-12), 120.4 (C-13), 114.5 (C-14), 113.4 (C-15), 166.4 (C-1′), 116.1 (C-2′), 157.6 (C-3′), 27.6 (C-4′), and 20.4 (C-5′). The above data were consistent with those reported in the literature (Dong Xueyun et al., 2014). Therefore, compound 1 was identified as ainsliaolide A.

[0080] Structural identification of compound 2:

[0081] See also Figures 3 to 11 Compound 2 is a colorless oil with the molecular formula C 20 H 24 O5 was detected by high resolution mass spectrometry HR-ESI-MS at m / z 367.1515 ([M+Na] + The calculated value was 367.1516) and the degree of unsaturation of the compound was further calculated to be 9. 1 H-NMR spectrum ( Figure 3 ) shows that in δ H There are two singlet methyl groups at 2.18 (3H, s, Me-5′) and 1.91 (3H, s, Me-4′); 5 olefin proton signals δ H6.27 (1H, d, J = 3.4 Hz, H-13), 5.69 (1H, s, 1.3 Hz, H-2′), 5.55 (1H, d, J = 3.0 Hz, H-13), 5.45 (1H, s, H-15) and 5.25 (1H, s, H-15). 13 C-NMR spectrum ( Figure 4 ) shows that the compound has 20 carbons, including 2 methyl groups, 6 quaternary carbons, 6 methine groups and 6 methylene groups. According to the above NMR data, 5 unsaturations are assigned to 2 carbonyl groups and 3 double bonds, which means that the structure of compound 2 is a tetracyclic sesquiterpene. There are slight differences between the NMR data of compound 2 and compound 1. Unlike compound 1, the NMR spectrum of compound 2 does not have the typical terminal double bond signal of C-14 of guaiacyl sesquiterpene lactone, but shows an additional methylene group (δ H 2.75,2.54;δ C 50.3). The methylene group at C-14 (δ C 50.3) was confirmed by the HMBC correlation signals of H-14 / C-10, H-14 / C-1 and H-14 / C-9. At the same time, in the HMBC spectrum, it can be observed that H-1 is correlated with C-2, C-3, C-4, C-5, C-6, C-9, C-10 and C-14; H-15 is correlated with C-3 and C-5; H-13 is correlated with C-7, C-11, C-12 ( Figure 9 ) indicates the presence of double bonds at C-4 (15) and C-11 (13). Two methyl signals (δ H 2.18, 3H, s, H-5′ and 1.91, 3H, s, H-4′) and one olefin proton signal (δ H 5.69, 1H, s, H-2′) indicates the presence of a senecioyloxy group. The correlation between H-3 and C-1′ indicates that the senecioyloxy group is attached to C-3. H-1 / H-3, H-1 / H-7, H-3 / H-5 and H-7 / H-9α[δ H 1.88-1.98 (m, 1H)] ROESY correlation signals indicate that H-1, H-3, H-5, and H-7 are α-oriented. Similarly, H-6 / H-14 and H-6 / H-9β(δ H 1.78) indicates that H-6 and CH2-14 are β-oriented ( Figure 10 Finally, the absolute configuration of compound 2 was determined by ECD calculation. The calculated ECD curve of compound 2 is very consistent with the measured curve ( Figure 11 Therefore, the absolute configurations of compound 2 were determined to be 1R, 3S, 5R, 6S, 7S, and 10R, and it was named ainslide C.

[0082] Example 1: Compound 1 and Compound 2 reduce the ratio of propidium iodide (PI)-positive cells and block cell pyroptosis

[0083] (1) Experimental methods:

[0084] J774A.1 cells were cultured at a density of 2×10 5 Cells were plated in 24-well plates and stimulated with 200 ng / mL LPS for 3.5 h. The supernatant was discarded and the corresponding substances were added and incubated for 30 min.

[0085] Then, 10 μM Nigericin was added for 1 h to induce inflammasome activation, the supernatant was discarded, and a premix of PI (3 μg / mL) and Hoechst33342 (0.5 μg / mL) diluted with PBS was added. After staining for about 15 minutes, the cells were observed under a fluorescent inverted microscope (100X). Five different fields of view were selected, and the PI-positive cells (cells releasing red fluorescent signals) in each field of view were counted and the average value was calculated.

[0086] PI positive rate = average number of cells with red fluorescent signal under microscopy / average number of total cells under microscopy × 100%

[0087] (2) The propidium iodide (PI) used in the above experiment is a fluorescent dye for detecting cell pyroptosis. It cannot pass through the intact cell membrane of living cells, but can pass through damaged cell membranes and embed into double-stranded DNA to release red fluorescent signals. In the above experiment, by adding activated inflammasomes, cells are induced to undergo pyroptosis and holes are formed in the cell membrane, allowing PI dye to enter and exit the cells freely. Cells undergoing pyroptosis will release red fluorescent signals, while cells that do not will not. The higher the PI positive rate calculated based on the results of microscopy, the higher the degree of pyroptosis of the treated cells.

[0088] The experimental results can be found in Figure 12 and Figure 13 As shown, Figure 12 and Figure 13 Compound 1 (10 μM) and Compound 2 (10 μM) significantly reduced the PI-positive rate of cells and inhibited cell pyroptosis. Compared with the model group, ***P<0.001 data showed significant efficacy.

[0089] Depend on Figure 12 and Figure 13 It can be seen that after the activated inflammasomes were mixed with the solutions of compound 1 and compound 2 of the present invention, the number of PI-positive cells was relatively small after treatment for 3 to 4 days, indicating that compound 1 (10 μM) and compound 2 (10 μM) inhibited cell pyroptosis and significantly reduced the PI-positive rate of cells (***P<0.001). Both compounds were able to reduce the PI-positive rate to below 19%.

[0090] Example 2 Compound 1 and Compound 2 inhibit the release of lactate dehydrogenase (LDH) and block cell pyroptosis experiment

[0091] (1) Experimental methods:

[0092] J774A.1 cells were cultured at a density of 2×10 5 Plate cells per well in a 24-well plate and culture overnight. Observe the cell plating and cell status the next day to determine if the experiment is suitable. If so, remove the supernatant and add 1 mL of Opti-MEM medium containing 200 ng / mL lipopolysaccharide (LPS) for sensitization. Incubate for 3.5 hours.

[0093] After the stimulation was completed, the supernatant was discarded and Opti-MEM medium containing compound 1 and compound 2 was added for 30 minutes.

[0094] After the drug treatment reached the desired time, cells were stimulated for 1 hour by adding Opti-MEM medium containing nigericin (an NLRP3 inflammasome inducer, final concentration: 10 μM). MCC950 was used as a positive control (MCC950 is a typical inhibitor of the NLRP3 inflammasome).

[0095] 100 μL of cell culture supernatant was collected and placed in a 96-well plate. Non-Radioactive Cytotoxicity Assay, Promega; USA), add the detection reagent, place at room temperature for 15 minutes, and place the 96-well plate in a microplate reader to read the OD 490 After the values ​​were calculated, the LDH release rate (%) was calculated according to the following formula.

[0096] LDH release rate = (OD 490 -OD in the normal group 490 ) / (OD of the model group 490 -OD in the normal group 490 )×100%

[0097] (2) Experimental results

[0098] Lactate dehydrogenase (LDH) is a metabolic enzyme normally present in cells. Living cells, due to their intact cell membranes, do not release LDH extracellularly. However, after pyroptosis occurs, the cell membrane is punctured, allowing LDH to be released extracellularly. The extent of pyroptosis can be assessed by measuring the rate of extracellular LDH release in cell culture medium supplemented with inflammasome inducers.

[0099] The above experimental results can be found in Figure 14 , Figure 14Compound 1 (10 μM) and compound 2 (10 μM) inhibited pyroptosis in J774A.1 cells and blocked the extracellular release of LDH. ***P < 0.001 compared with the model group.

[0100] The results are as follows Figure 14 As shown in the figure, the LDH release rates of compound 1 and compound 2 are much lower than that of the model group, indicating that the addition of the compound can protect the cells, inhibit the activation of inflammatory factors, effectively reduce the proportion of cell pyroptosis, inhibit cell pyroptosis, and block the extracellular release of LDH.

[0101] Example 3: Compound 1 inhibits the formation of GSDMD-NT and blocks cell pyroptosis

[0102] (1) Experimental methods:

[0103] J774A.1 cells were cultured at a density of 2×10 5 Plate cells / well in a 24-well plate and culture overnight. Observe the cell laying and cell status the next day to determine whether the experiment can be carried out. If the supernatant can be removed, add 1 mL of Opti-MEM medium containing 200 ng / mL lipopolysaccharide (LPS) for sensitization for 3.5 hours.

[0104] After the stimulation was completed, the supernatant was discarded and Opti-MEM medium containing each treatment compound was added for 30 minutes. After the drug stimulation reached the time, Opti-MEM medium containing NLRP3 inflammasome inducer (Nigericin, final concentration of 10 μM) was added to stimulate the cells for 1 hour.

[0105] Add 200 μL of RIPA lysis buffer to each well, incubate on ice for 30 minutes, and sonicate for 1 minute. Add 6X Loading Buffer and incubate in a boiling water bath for 5 minutes. Perform SDS-PAGE electrophoresis, and perform immunoblotting using conventional methods. The primary antibody used was anti-GSDMD rabbit monoclonal antibody (ab209845; Abcam). β-Tubulin was used as the internal control protein.

[0106] (2) Experimental results:

[0107] Pyroptosis is a specialized form of programmed cell death. Activation of the NLRP3 inflammasome typically leads to activated Caspase-1, which cleaves the GSDMD protein, activating its N-terminus to form the GSDMD-NT protein. The GSDMD-NT protein then creates pores in the cell membrane, triggering pyroptosis.

[0108] The results of the above experiments are as follows Figure 15 As shown, Figure 15Middle: The concentration of compound 1 was 10 μM. Nigericin-induced J774A.1 cell pyroptosis model was used. Figure 15 Western blot analysis showed that compound 1 could reduce the cleavage of GSDMD and the generation of GSDMD-NT in a dose-dependent manner.

[0109] Example 4: Compound 1 and Compound 2 inhibit Caspase-1 and IL-1β protein cleavage and block inflammasome activation

[0110] (1) Experimental methods:

[0111] J774A.1 cells were cultured at a density of 2×10 5 Cells were plated at 100 cells / well in a 24-well plate and cultured overnight. The next day, the cell plating and cell status were observed to determine whether the experiment could be performed. If possible, the supernatant was removed and 1 mL of Opti-MEM medium containing 200 ng / mL lipopolysaccharide (LPS) was added for sensitization for 3.5 hours. After stimulation, the supernatant was discarded and Opti-MEM medium containing the respective treatment compound was added for 30 minutes. After the drug stimulation reached the desired time, Opti-MEM medium containing the NLRP3 inducer (Nigericin, final concentration 10 μM) was added to stimulate the cells for 1 hour.

[0112] 500 μL of cell culture supernatant was collected and protein was concentrated using the conventional trichloroacetic acid (TCA) method. 2× SDS loading buffer was then added, vortexed, and heated at 100°C for 10 min. SDS-PAGE electrophoresis was performed, and immunoblotting was performed using conventional methods. The primary antibody used was a mouse monoclonal anti-Caspase-1 antibody (AG-20B-0042-C100; Adipogen). The primary antibody used was a goat polyclonal anti-IL-1β antibody (AF-401; R&D). β-actin was used as the internal control protein.

[0113] (2) After the NLRP3 inflammasome is activated, pro-Caspase-1 is activated to Caspase-1; and because cell pyroptosis destroys the integrity of the cell membrane, cleaved Caspase-1 (also known as Cleaved Caspase-1) can be detected in the cell culture supernatant for treatments that cause cell pyroptosis. Therefore, detecting the expression level of Cleaved Caspase-1 in the culture supernatant can reflect the inhibitory effect of the compound on cell pyroptosis. Since Caspase-1 is essentially a protease, it can hydrolyze specific sites of pro-IL-1β, leading to the maturation of IL-1β and the formation of mature IL-1β. IL-1β is an important pro-inflammatory cytokine in the body, and its large-scale maturation often leads to the occurrence of inflammatory factor storms.

[0114] The experimental results can be found in Figure 16 and Figure 17 The concentrations of compound 1 and compound 2 in each treatment were 10 μM. The Nigericin-induced J774A.1 cell pyroptosis model was used. Both compound 1 and compound 2 can inhibit the activation of Pro-Caspase-1 to Caspase-1 in cells in a dose-dependent manner, indicating that both compound 1 and compound 2 can inhibit the cleavage of Caspase-1 in the cell pyroptosis model, thereby inhibiting the occurrence of inflammasome activation from the source. Both compound 1 and compound 2 can inhibit the activation of Pro-IL-1β to IL-1β in cells in a dose-dependent manner, indicating that both compound 1 and compound 2 can effectively inhibit the maturation of IL-1β, block inflammasome activation, and thus inhibit the occurrence of inflammatory cytokine storm.

[0115] Example 5: Compound 1 inhibits IL-1β secretion and blocks inflammasome activation

[0116] (1) Experimental methods:

[0117] J774A.1 cells were cultured at a density of 2×10 5 Cells were plated at 100 cells / well in a 24-well plate and cultured overnight. The next day, the cell plating and cell status were observed to determine whether the experiment could be performed. If the supernatant can be removed, 1 mL of Opti-MEM medium containing 200 ng / mL lipopolysaccharide (LPS) was added for sensitization for 3.5 hours. After stimulation, the supernatant was discarded and Opti-MEM medium containing the respective treatment compound was added for 30 minutes. After the drug stimulation reached the desired time, Opti-MEM medium containing the NLRP3 inducer (Nigericin, final concentration 10 μM) was added to stimulate the cells for 1 hour.

[0118] 100 μL of cell culture supernatant was collected and commercial ELISA kits (DY401, DY008B; R&D Company) were used to detect the inhibition of IL-1β secretion by the compounds. Standard curve equation: OD value = 0.0021 × protein concentration (pg / mL) + 0.0801

[0119] In this example, compound 1 was diluted 2-fold in each treatment, with a final concentration gradient of 2.5 μM, 5 μM, and 10 μM. Nigericin-induced J774A.1 cell pyroptosis model was used, and the culture supernatant of each treatment was collected for ELSIA analysis. IL-1β secretion was calculated based on the standard curve.

[0120] (2) Experimental results are as follows Figure 18 As shown, solutions of compound 1 at different concentrations can inhibit the secretion of IL-1β and block the activation of inflammasomes, and the inhibitory effect is dose-dependent.

[0121] Example 6: Compound 1 inhibits the formation of ASC oligomers (ASC Oligomerization) and blocks the formation of NLRP3 inflammasomes

[0122] (1) Experimental methods:

[0123] Cells were plated in a biosafety cabinet and the cells were plated in 6-well plates with a cell number of 1×10 6 Cells were cultured overnight in a cell culture incubator, and the cell status was observed the next day. The cells were stimulated to produce NLRP3 inflammasomes. All cell supernatants were removed, and 300 μL / well of non-denaturing cell lysis buffer (containing 0.5% Triton-X-100, 1% PMSF) prepared in 1×PBS was added. The cells were repeatedly pipetted and aspirated until the cells were fully lysed. The lysed cell fluid was transferred to an EP tube and centrifuged at 4°C 6000×g for 10 minutes. The supernatant was retained as a protein internal reference sample, and 300 μL / tube of non-denaturing cell lysis buffer was added to the precipitate, and the precipitate was centrifuged at 4°C 6000×g for 10 minutes. The supernatant was discarded and the precipitate was retained. Non-denaturing cell lysis buffer containing 2 mM DSS was added to the precipitate, and the precipitate was rotated on a disc rotator at room temperature for 1 hour, during which the liquid was continuously flipped in the EP tube.

[0124] The cells were centrifuged at 6000 × g at 4°C for 10 min, and all supernatants were removed. 30 μL / tube of 2× Laemmli buffer was added. After vortex centrifugation, the cells were boiled in a boiling water bath for 10 min, ice-shocked for 10 min, and stored in a refrigerator at -20°C. Samples were prepared for immunoblot analysis.

[0125] (2) Experimental results:

[0126] See also Figure 19The formation of ASC oligomers is a prerequisite for the activation of Pro-Caspase-1. Therefore, detecting the oligomerization of ASC is an important means to verify the activation of NLRP3 inflammasome. The DSS (disuccinimidyl suberate) used in this article is an indestructible membrane-permeable cross-linker. Its two ends contain two amine-reactive N-hydroxysuccinimide (NHS) ester structures with the same structure and function. Under appropriate pH conditions, this structure can bind to the primary amine and structure of the protein to form a stable amide bond. Using this property of DSS, oligomerized ASC is cross-linked to observe the effect of compound 1 on ASC polymerization. The results after cross-linking are analyzed by Western blot, as shown in Figure 2. Figure 19 As shown, compound 1 attenuated the formation of ASC monomers, dimers, and oligomers in a dose-dependent manner. These results indicate that when compound 1 inhibits NLRP3 inflammasome activation, it also blocks NLRP3 inflammasome-induced ASC oligomerization.

[0127] Example 7: Compound 1 inhibits MSU-induced mitochondrial damage in J774A.1 cells

[0128] (1) Experimental methods:

[0129] Cells were plated in a biosafety cabinet and the cells were plated in 6-well plates with a cell number of 1×10 6 Cells were cultured overnight in a cell culture incubator. The cells were observed the next day and stimulated with MSU (150 μg / mL) crystals to produce NLRP3 inflammasomes. All cell supernatants were removed, the cells were washed twice with 1× PBS, and 200 μL / well of a 5 μg / mL JC-1 dye solution was added. The cells were incubated in a cell culture incubator for 15 min and photographed using a fluorescence inverted microscope.

[0130] (2) Experimental results:

[0131] See also Figure 20 The effect of compound 1 on mitochondrial damage in MSU-induced J774A.1 cells was detected, which can reflect the protective effect of the compound on mitochondria. JC-1 is used as a fluorescent probe to detect mitochondrial membrane potential. Normal mitochondrial membrane potential is high. JC-1 polymerizes in the mitochondrial matrix to produce a red fluorescent signal. However, the damaged mitochondrial matrix of damaged cells cannot polymerize with JC-1, and the unpolymerized JC-1 will emit a green fluorescent signal. After JC-1 staining, fluorescence microscopy analysis was performed, and the results were as follows: Figure 20 As shown in the results, compound 1 treatment significantly reduced the MSU-induced mitochondrial membrane potential loss, indicating that compound 1 can alleviate mitochondrial dysfunction during NLRP3 inflammasome activation in a dose-dependent manner.

[0132] Example 8: Compound 1 inhibits the production of mitochondrial reactive oxygen species (mt-SOX) in J774A.1 cells induced by MSU

[0133] (1) Experimental methods:

[0134] Cell plating and cell stimulation to produce NLRP3 inflammasomes were performed in the same manner as JC-1 staining. MitoSOX dye was diluted to 5 μM with 1× PBS, and then 1000× Hoechest 33343 dye was added. The dye solution was added to the cell culture plate at 200 μL / well, incubated in a cell culture incubator for 15 min, and observed and photographed using an inverted fluorescence microscope.

[0135] (2) Experimental results:

[0136] See also Figure 21 , mt-ROS is the main mediator of NLRP3 inflammasome activation. Detecting the effect of compound 1 on the production of mitochondrial reactive oxygen species in MSU-induced J774A.1 cells can reflect the compound's effect on reducing mitochondrial reactive oxygen species.

[0137] MitoSOX is a mitochondrial superoxide indicator that can be targeted to mitochondria and relies on mtmP to accumulate in mitochondria. After oxidation, it binds to mitochondrial DNA and emits red fluorescence. It is used to detect the production of mt-ROS in living cells. Immunofluorescence analysis showed that after LPS and MSU stimulated J774A.1 cells to activate inflammasomes, compound 1 could reduce the production of mt-ROS (see Figure 21 ).

[0138] Example 9: Compound 1 inhibits joint swelling in mice with gouty arthritis induced by sodium urate crystals (MSU)

[0139] (1) Experimental methods:

[0140] Compound preparation: The compound was dissolved in PBS solution containing 0.25% Tween-80, and the low concentration was 25 mg / kg, and the high concentration was 50 mg / kg. The solvent was PBS solution containing 0.25% Tween-80.

[0141] Animal Grouping: Twenty 8-week-old KM male mice were divided into five groups of four mice each: normal group, model group, low-concentration Compound 1 group, and high-concentration Compound 1 group. This study was approved by the Yunnan University Laboratory Animal Welfare and Ethics Committee and complies with ethical standards (Approval No.: YNUCARE20210056).

[0142] Compound 1 was administered intraperitoneally to mice at high and low concentrations, with 1 mg and 0.5 mg of compound 1 dissolved in 300 μL of saline. The mice were then anesthetized with 350 mg / kg of tribromoethanol intraperitoneally until they were fully anesthetized. The blank control group was injected with 200 μL of saline. MSU crystals were prepared in saline to a 1 mg / 20 μL solution. 20 μL of the MSU solution was injected into the joints of mice in the model and treatment groups using a microsyringe. The blank control group was injected with 20 μL of saline. After 24 hours of waiting, mice were sacrificed by cervical dislocation the next day. Joints were photographed and joint diameters were measured with a vernier caliper. Joint tissue was then collected. Mice were disinfected by soaking in 75% alcohol and then minced into small pieces using medical scissors. The pieces were then homogenized in RIPA lysis buffer containing protease inhibitors using an electric homogenizer. After incubation on ice for 20 minutes, the cells were centrifuged at 10,000 × g for 10 minutes at 4°C. The cell supernatant was collected and added with 6× SDS Sample Buffer. After vortexing and centrifugation, the samples were boiled in a boiling water bath for 10 minutes, ice-shocked for 10 minutes, and stored at -20°C for Western blotting. Furthermore, joint tissue was collected, cut into small pieces, and homogenized in 100 μL of Opti-MEM using an electric homogenizer. Opti-MEM was then added to 400 μL. The samples were transferred to a culture dish and incubated overnight in a cell culture incubator. The supernatant was then collected and analyzed by ELISA.

[0143] (2) Experimental results

[0144] See also Figure 22 、 Figure 23 、 Figure 24 and Figure 25 Gout is characterized by edema and is defined as pain occurring in the first metatarsophalangeal joint. The onset of gout seriously affects people's normal lives and brings great distress. Existing gout treatments, such as nonsteroidal anti-inflammatory drugs (NSAIDs), colchicine, or corticosteroids, can be very harmful to patients with gout-related complications. Therefore, we urgently need to find a safe and targeted new drug to treat patients with gouty arthritis. Gout patients are often accompanied by excessive activation of inflammation. MSU entering the joints can cause NLRP3 inflammasome-dependent arthritis in KM mice. Therefore, compound 1, which has an inhibitory effect on the NLRP3 inflammasome, has the potential to treat gouty arthritis.

[0145] It is necessary to investigate the effect of compound 1 on MSU-induced gouty arthritis in KM mice. Compared with the model group, the ankle swelling of mice injected intraperitoneally with compound 1 at concentrations of 25 mg / kg and 50 mg / kg was significantly alleviated, and the ankle swelling of mice injected with higher concentrations was more significantly alleviated (see Figure 22 and Figure 23 In addition, the administration inhibited the cleavage of Pro-Caspase-1 and the maturation of IL-1β in the mouse arthritis model in a dose-dependent manner, but did not affect the in vivo expression levels of NLRP3 and ASC proteins (see Figure 24 Finally, compound 1 reduced the expression level of IL-1β in the joint culture fluid of a mouse arthritis model and inhibited the production of IL-1β in a dose-dependent manner (see Figure 25 ).

[0146] These results indicate that compound 1 can treat MSU-mediated acute gouty arthritis in mice by inhibiting the production of IL-1β and Caspase-1.

[0147] Compounds 1 and 2 of the present invention inhibit inflammasome activation and block pyroptosis. The sesquiterpenoid compounds of the present invention are used in the preparation of products for preventing or treating NLRP3 inflammasome-related diseases. The NLRP3 inflammasome-related diseases are at least one of gout, arthritis, alcoholic liver damage, non-alcoholic liver damage, atherosclerosis, type II diabetes, sepsis, multiple sclerosis, systemic lupus erythematosus, Alzheimer's disease, Parkinson's disease, silicosis, cold pyrin syndrome, acne, psoriasis, dandruff, tinea versicolor, seborrheic dermatitis, UV-induced skin damage, UV-induced tanning, UV-induced skin aging, and insect bites. The dosage form of the product is at least one of tablets, dispersible tablets, lozenges, orally disintegrating tablets, sustained-release tablets, capsules, soft capsules, pills, granules, injections, powders for injection or aerosol powders, films, plasters, patches, creams, liniments, creams, and lotions. Products include medicines, health foods, cosmetics, and sanitary disinfectants. Cosmetics are any of shampoo, shampoo cream, cooling oil, wind oil essence, toner, floral water, face cream, face mask, face gel, sunscreen, sunscreen gel, and sunscreen lotion.

[0148] In summary, the sesquiterpenoid compound extracted and separated from the axillary wormwood and its application of the present invention, the sesquiterpenoid compound is at least one of compound 1 and compound 2, the molecular formula of compound 1 is C 20 H 24 O4, the structural formula is shown in formula 1; the molecular formula of compound 2 is C 20 H 24O5, the structural formula is shown in Formula 2. Compounds 1 and 2 of the present invention have the effect of inhibiting inflammasome activation and blocking cell pyroptosis, and can be used in the preparation of inflammasome inhibitors, the treatment of gouty arthritis, and the preparation of drugs, health foods, cosmetics, and sanitary disinfectants for preventing, treating, or improving diseases related to cell pyroptosis.

[0149] Those skilled in the art should recognize that the above embodiments are merely intended to illustrate the present invention and are not intended to limit the present invention. As long as they are within the spirit of the present invention, any changes or modifications to the above embodiments will fall within the scope of the claims of the present invention.

Claims

1. A sesquiterpenoid compound extracted and separated from the herb Thunbergia axillariae, characterized in that: The sesquiterpenoid compound is compound 2; the molecular formula of compound 2 is C 20 H 24 O5, the structural formula is shown in formula (2):

2. The sesquiterpenoid compound extracted and separated from the Herba Lysimachiae as claimed in claim 1, characterized in that: The steps of extracting and separating the sesquiterpenoid compounds from the hyacinthus annuus are as follows: (1) The dried whole herb of Ainsliaea pertyoides Franch. was extracted with 85% methanol, and then concentrated and dried under reduced pressure to obtain an 85% methanol extract; (2) dissolving the 85% methanol extract obtained in step (1) in water, extracting with ethyl acetate which is immiscible with water, and recovering the solvent under reduced pressure to obtain an extract; (3) The extract obtained in step (2) was separated by D-101 macroporous resin column chromatography using a methanol / water (50:50-100:0, v / v) gradient elution to obtain five fractions (Fr.1-Fr.5); (4) Fr.2 obtained in step (3) above was separated by silica gel column chromatography, and eluted with a gradient of chloroform / methanol (10:1-0:1, v / v) to obtain 9 fractions (Fr.2.1-Fr.2.9); Fr.3 was separated by silica gel column chromatography, and eluted with a gradient of petroleum ether / ethyl acetate (30:1-1:1, v / v) to obtain 6 fractions (Fr.3.1-Fr.3.6); (5) Fr.2.8 and Fr.3.3 obtained in the above step (4) were separated by semi-preparative HPLC chromatography, eluted with 70% and 60% acetonitrile, respectively, to obtain compound 2.

3. A use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 1, characterized in that: The sesquiterpenoid compound is used in the preparation of products that inhibit inflammasome activation and / or block cell pyroptosis.

4. The use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 3, characterized in that: The inflammasome is the NLRP3 inflammasome.

5. The use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 3, characterized in that: The inhibition of inflammasome activation is at least one of inhibiting the cleavage of Caspase-1 protein in inflammasome-inducing cells, inhibiting the maturation of IL-1β in inflammasome-inducing cells, and inhibiting the secretion of IL-1β in inflammasome-inducing cells.

6. The use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 5, characterized in that: The sesquiterpenoids inhibit the secretion of IL-1β in inflammasome-induced cells. 50 All were less than 10μM.

7. The use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 3, characterized in that: The blocking of cell pyroptosis is at least one of reducing the propidium iodide positive rate of inflammasome-induced cells, inhibiting the release of lactate dehydrogenase in inflammasome-induced cells, and inhibiting the formation of GSDMD-NT in inflammasome-induced cells.

8. The use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 7, characterized in that: When the concentration of the sesquiterpenoid compound is 10 μM, the propidium iodide positive inhibition rate is 15-19%, and the lactate dehydrogenase release rate is 7-13%.

9. A use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 1, characterized in that: The use of the sesquiterpenoid compound in the preparation of a product for preventing or treating NLRP3 inflammasome-related diseases.

10. The use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 9, characterized in that: The NLRP3 inflammasome-related disease is at least one of gout, arthritis, alcoholic liver damage, non-alcoholic liver damage, atherosclerosis, type II diabetes, sepsis, multiple sclerosis, systemic lupus erythematosus, Alzheimer's disease, Parkinson's disease, silicosis, cold pyrin syndrome, acne, psoriasis, dandruff, tinea versicolor, seborrheic dermatitis, UV-induced skin damage, UV-induced skin tanning, UV-induced skin aging, and insect bites.

11. The use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 9, characterized in that: The dosage form of the product is at least one of tablets, dispersible tablets, lozenges, orally disintegrating tablets, sustained-release tablets, capsules, soft capsules, pills, granules, injections, powder injections or aerosol powders, films, plasters, patches, emulsions, liniments, creams and lotions.

12. The use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 9, characterized in that: The products include medicines, health foods, cosmetics and sanitary disinfection products.

13. The use of the sesquiterpenoid compounds extracted and separated from the Herba Lysimachiae as claimed in claim 12, characterized in that: The cosmetic is any one of shampoo, shampoo cream, cooling oil, wind oil, toner, floral water, facial cream, facial mask, facial gel, sunscreen milk, sunscreen cream, sunscreen gel and sunscreen liquid.

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