Compound with anti-inflammatory effect in Enshi phellodendron amurense, separation and application

Abstract

The invention discloses a compound with an anti-inflammatory effect in Enshi Sichuan amur corktree, separation and application, and belongs to the technical field of medicines. The invention particularly discloses a compound which is separated from Enshi chuan amur corktree and has an anti-inflammatory effect, a separation preparation method and application. Researches prove that the compound has an inhibition effect on the expression of inflammatory factors IL-1beta and IL-17A, can inhibit the generation and secretion of an inflammatory factor IL-6 in Th17 cells, can reduce the expression level of Caspase-1 and NLRP3 proteins in colon tissues of mice, and can reduce the inflammatory response of ulcerative colitis of DSS modeling mice in a dose-dependent manner. The compound (limonin compound) with the anti-inflammatory effect in Enshi Huangchuan amur corktree can be used as a lead compound for development of anti-inflammatory drugs, and is of great significance to comprehensive utilization and development of Enshi Huangchuan amur corktree.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and particularly relates to compounds with anti-inflammatory effects in Enshi Phellodendron chinense Schneid., separation, and applications thereof. Background Art

[0002] Ulcerative colitis (UC) is a disease mainly characterized by chronic and recurrent intestinal inflammation. Patients often have symptoms such as abdominal pain, diarrhea, hematochezia, anemia, etc. In severe cases, complications such as intestinal perforation and intestinal obstruction may also occur. It belongs to one of the inflammatory bowel diseases and is commonly known as the "green cancer". Ulcerative colitis has a long course, is difficult to cure, and is prone to canceration. It has been listed as one of the modern intractable diseases by the World Health Organization and has become a global disease, seriously threatening the lives and health of people around the world.

[0003] Phellodendron bark is one of the important traditional Chinese medicines in China and has significant anti-inflammatory and detoxifying effects. Among them, Sichuan Phellodendron bark is the dried bark of Phellodendron chinense Schneid. of the genus Phellodendron in the Rutaceae family. It is mainly distributed in the Wuling Mountain planting area of Hubei Province, including 7 counties and cities such as Enshi, Lichuan, Xianfeng, Hefeng, and Jianshi, and is one of the 13 famous and high-quality Chinese medicinal materials that Hubei Province focuses on developing. Sichuan Phellodendron bark is cold in nature and bitter in taste, and has the effects of clearing heat and detoxifying, purging fire and drying dampness. Clinically, it is commonly used to treat inflammatory diseases such as acute enteritis, acute bacillary dysentery, and urinary tract infections. Topically, it can treat otitis media, acute conjunctivitis, and burns.

[0004] At present, the research on limonoid compounds at home and abroad is relatively extensive, but there are few reports on the method of extracting limonoid compounds from Phellodendron chinense Schneid. produced in Enshi, and existing methods mostly use traditional extraction processes, which have problems such as large solvent consumption and low extraction efficiency. Summary of the Invention

[0005] The purpose of the present invention is to provide limonoid compounds with anti-inflammatory effects in Enshi Phellodendron chinense Schneid. and their separation methods, and to clarify the application value of the limonoid compounds in the preparation of anti-inflammatory drugs, thereby solving the problem of many adverse effects of existing drugs.

[0006] According to the first aspect of the present invention, a tetranortriterpenoid compound is provided, and the structural formula of the tetranortriterpenoid compound is shown as follows:

[0007]

[0008]

[0009]

[0010] Among them, R in formula VII is selected from hydroxy or methoxy; R in formula VIII is selected from α-hydroxy or β-hydroxy; R1 and R2 in formula IX are each independently selected from hydroxy or carbonyl; R1 and R2 in formula X are each independently selected from α-hydrogen, β-hydrogen, hydroxy or methoxy; R in formula XI is selected from hydroxy or hydrogen; R1 and R2 in formula XIV are each independently selected from hydroxy or carbonyl.

[0011] Preferably, formula I is compound 1, formula II is compound 2, formula III is compound 3, formula IV is compound 4, formula V is compound 5, formula VI is compound 6, formula VII is compound 7 or compound 8, formula VIII is compound 9 or compound 10, formula IX is compound 11 or compound 12, formula X is compound 13, compound 14 or compound 15, formula XI is compound 16 or compound 17, formula XII is compound 18, formula XIII is compound 19, formula XIV is compound 20 or compound 21, formula XV is compound 22, formula XIV is compound 23;

[0012]

[0013]

[0014]

[0015] On the other hand, according to the present invention, there is provided a method for separating the tetranortriterpenoid compounds, comprising the following steps:

[0016] (1) Dry the bark of Enshi Phellodendron chinense Schneid., pulverize it, extract with ethanol, and concentrate under reduced pressure to obtain a total extract.

[0017] (2) Suspend the total extract obtained in step (1) in water, extract with chloroform and ethyl acetate respectively, and then concentrate under reduced pressure to obtain chloroform and ethyl acetate part extraction extracts respectively.

[0018] (3) Dissolve the chloroform part extraction extract obtained in step (2) with a mixed solvent of dichloromethane and methanol, mix with silica gel and then load it onto a silica gel column, perform gradient elution with a dichloromethane and methanol system, and combine with normal phase thin layer chromatography TLC color development to be divided into 12 components A1 - A12; dissolve the ethyl acetate part extraction extract obtained in step (2) with a mixed solvent of dichloromethane and methanol, mix with silica gel and then load it onto a silica gel column, perform gradient elution with a dichloromethane and methanol system, and combine with normal phase thin layer chromatography TLC color development to be divided into 8 components B1 - B8;

[0019] When component A3 was separated by gradient elution through an ODS reversed-phase column chromatography to obtain 17 sub-components A3a - A3q; among them, A3l was separated by gel column chromatography into 5 sub-components A3l1 - A3l5, and among them, A3l2 was further purified by semi-preparative HPLC to obtain compound 19;

[0020] When component A4 was separated by gradient elution through an ODS reversed-phase column chromatography to obtain 16 sub-components A4a - A4p; among them, A4f was separated by a gel column into 3 sub-components A4f1 - A4f3, and among them, A4f3 was separated by normal-phase column chromatography to obtain 4 sub-components A4f3a - A4fd, and A4fd was further purified by semi-preparative HPLC to obtain compound 21 and compound 20;

[0021] When component A4h was separated by gel column chromatography into 3 sub-components A4h1 - A4h3, and among them, A4h3 was recrystallized to obtain compound 16;

[0022] When component A4i was separated by gel column chromatography into 3 sub-components A4i1 - A4i3, and among them, A4i3 was purified by semi-preparative HPLC to obtain compound 18;

[0023] When component A5 was separated by gradient elution through an ODS reversed-phase column chromatography to obtain 14 sub-components A5a - A5n; among them, A5h was separated by a normal-phase silica gel column into 5 sub-components A5h1 - A5h5, and among them, A5h3 was recrystallized to obtain compound 17;

[0024] When component A5j was separated using a normal-phase silica gel column to obtain 10 sub-components A5j1 - A5j10; among them, A5j5 was separated by gel column chromatography into 3 sub-components A5j5a - A5j5c, and among them, A5j5a was recrystallized to obtain compound 9;

[0025] When component A5j5c was separated by semi-preparative HPLC to obtain compound 10;

[0026] When component A5j7 was separated by gel column chromatography to obtain compound 22;

[0027] When component A7 was separated by normal-phase silica gel column chromatography combined with TLC color development to obtain 9 sub-components A7a - A7i; A7e was separated by gel column chromatography into 3 sub-components A7e1 - A7e3, and among them, A7e2 was further separated by a normal-phase silica gel column into 5 sub-components A7e2a - A7e2e, and A7e2e was recrystallized to obtain compound 3;

[0028] When component A7f was separated by a normal-phase silica gel column to obtain 5 sub-components A7f1 - A7f5, among which A7f3 was separated by a reverse-phase medium-pressure column to obtain 6 sub-components A7f3a - A7f3f, and A7f3f was purified by semi-preparative HPLC to obtain compound 11 and compound 12;

[0029] When component A9 was separated by reverse-phase medium-pressure column chromatography combined with TLC color development to obtain 19 sub-components A9a - A9s; among which A9j was separated by gel column chromatography to obtain 3 sub-components A9j1 - A9j3, and A9j2 was purified by a normal-phase silica gel column to obtain compound 13;

[0030] When component A9m was separated by gel column chromatography into 4 sub-components A9e1 - A9e4, among which A9e2 and A9e3 were combined and the main components were purified by a normal-phase silica gel column and a gel column respectively, and then further purified by semi-preparative HPLC to obtain compound 6;

[0031] When components A10 - A11 were combined and dissolved in pure methanol, the chlorophyll components were removed by an MCI column, and then gradient elution was performed using an ODS reverse-phase column chromatography to obtain 17 sub-components A10a - A10q; among which A10i was separated by gel column chromatography into 2 sub-components A10i1 - A10i2, and A10i1 was further purified by a normal-phase silica gel column and semi-preparative HPLC to obtain compound 1;

[0032] When component B4 was separated by ODS reverse-phase column chromatography combined with the TLC color development situation to obtain 12 sub-components B4a - B4l; among which B4h was first purified by a gel column to obtain the main component, and then further purified by HPLC to obtain compound 5;

[0033] When component B4i was purified by gel column chromatography and semi-preparative HPLC to obtain compound 8;

[0034] When component B4l was separated by gel column chromatography to obtain 4 sub-components B4l1 - B4l4, according to the TLC color development results, B4l2 was separated by a normal-phase silica gel chromatography column to obtain 7 sub-components B4l2a - B4l2g, and B4l2c was purified by semi-preparative HPLC to obtain compound 14 and compound 15;

[0035] When component B5 was separated by ODS reverse-phase column chromatography to obtain 9 sub-components B5a - B5i; among which B5d was separated by a gel column into 2 sub-components B5d1 - B5d2, and B5d2 was separated by a normal-phase silica gel column combined with the TLC color development situation to obtain 9 sub-components B5d2a - B5d2g, and B5d2f was further purified by semi-preparative HPLC to obtain compound 2;

[0036] When component B6 was separated by gradient elution through ODS reversed-phase column chromatography to obtain 7 sub-components B6a - B6g; among them, B6d was separated by gel column to obtain 2 sub-components B6d1 - B6d2, among which component B6d2 was separated by medium-pressure reversed-phase column to obtain 6 sub-components B6d2a - B6d2f, and among them, B6d2d was purified by semi-preparative HPLC to obtain compound 23 and compound 7;

[0037] When component B7 was separated by ODS reversed-phase column chromatography to obtain 9 sub-components B7a - B7i; among them, B7e was separated by normal-phase column chromatography into 7 sub-components B7e1 - B7e7, and among them, B7e6 was repeatedly purified by gel to obtain the main component, and then monomer compound 4 was obtained by semi-preparative HPLC.

[0038] According to another aspect of the present invention, there is provided the use of the tetranortriterpenoid compounds in the preparation of anti-inflammatory drugs.

[0039] Preferably, the anti-inflammatory drug is a drug for treating pneumonia, a drug for treating hepatitis or a drug for treating ulcerative colitis.

[0040] Preferably, the tetranortriterpenoid compounds are used to inhibit the expression of inflammatory factors IL-1β and IL-17A.

[0041] Preferably, the tetranortriterpenoid compounds are used to inhibit the production and secretion of inflammatory factor IL-6 in Th17 cells.

[0042] Preferably, the tetranortriterpenoid compounds are used to reduce the expression levels of Caspase-1 and NLRP3 proteins in colon tissues.

[0043] Generally speaking, compared with the prior art through the above technical solutions conceived by the present invention, the following technical advantages are mainly possessed:

[0044] (1) Compounds 1 - 8 provided in the present invention are all new compounds first prepared and discovered from Phellodendron chinense Schneid. var. wilsonii (Hemsl.) Huang in Enshi. Compound 1 is 1,4:14,15-diepoxy-7,8-seco-3,19:16,17-dilactone limonin. Compounds 2 - 3, 9 - 12, 16 - 17 are 1,4:14,15-diepoxy-3,19:16,17-dilactone limonin. Compound 4 is 1,4:14,15-diepoxy-3,19-lactone limonin. Compounds 5, 13 - 15 are 1,4:14,15-diepoxy-16,17-lactone limonin. Compounds 6, 19 - 22 are 14,15-epoxy-3,4:16,17-dilactone limonin. Compounds 7 - 8, 23 are 14,15-epoxy-16,17-lactone limonin. Compound 18 is 4,19-epoxy-3,19:16,17 dilactone limonin.

[0045] (2) The compounds provided by the present invention have an inhibitory effect on the expression of inflammatory factors IL-1β and IL-17A. At the same time, they can inhibit the production and secretion of the inflammatory factor IL-6 in Th17 cells, and reduce the expression levels of Caspase-1 and NLRP3 proteins in the colon tissues of mice; and dose-dependently reduce the inflammatory response of ulcerative colitis in mice modeled with DSS.

[0046] (3) Preferably, the compounds 9 and 13 provided by the present invention have a significant anti-inflammatory effect on lipopolysaccharide (LPS)-induced RAW264.7 cells, equivalent to that of the positive drug dexamethasone.

[0047] (4) Preferably, compounds 9 and 13 have a significant protective effect in a mouse model of acute ulcerative colitis induced by DSS, and the protective effect is stronger than that of the positive drug 5-aminosalicylic acid. The results of pathological sections of colon tissues show that most of the mucosal epithelium in the colon sections proliferates and repairs, the infiltration of inflammatory cells is significantly reduced, the number of glandular cells and goblet cells increases, and the arrangement is relatively neat. The degree of protection of the glandular structure is better than that of the positive drug 5-aminosalicylic acid. Description of the Drawings

[0048] Figure 1 It is a comparison diagram of the ECD calculation and measurement of compound 1.

[0049] Figure 2 It is a comparison diagram of the X-ray single crystal diffraction pattern of compound 2 and the ECD calculation and measurement.

[0050] Figure 3 It is a comparison diagram of the ECD calculation and measurement of compound 3.

[0051] Figure 4 It is a comparison diagram of the ECD calculation and measurement of compound 4.

[0052] Figure 5 It is a comparison diagram of the ECD calculation and measurement of compound 5.

[0053] Figure 6 It is a comparison diagram of the X-ray single crystal diffraction pattern of compound 6 and the ECD calculation and measurement.

[0054] Figure 7 It is a comparison diagram of the Boltzmann distribution prediction map of compound 7 and the ECD calculation and measurement.

[0055] Figure 8 It is a comparison diagram of the ECD calculation and measurement of compound 8.

[0056] Figure 9 It is the X-ray single crystal diffraction pattern of compound 9.

[0057] Figure 10It is the X-ray single crystal diffraction pattern of Compound 12.

[0058] Figure 11 It is the X-ray single crystal diffraction pattern of Compound 17.

[0059] Figure 12 It is the X-ray single crystal diffraction pattern of Compound 18.

[0060] Figure 13 It is the X-ray single crystal diffraction pattern of Compound 19.

[0061] Figure 14 It is the graph of the anti-ulcerative colitis activity results of Compounds 9 and 13 on mice.

[0062] Figure 15 It is the graph of the molecular docking results of Compounds 9 and 13. Detailed implementation manners

[0063] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0064] The present invention detects the chloroform and ethyl acetate extraction parts through a thin-layer chromatography plate, and combines the sulfuric acid-ethanol and Ehrlich color reactions, and finds that this part is rich in various limonoid components. For the separation and purification of this part, the present invention provides an efficient and environmentally friendly extraction method, which can significantly reduce costs and increase the yield of target compounds, providing a new technical approach for the development and utilization of limonoid compounds in Phellodendron chinense Schneid.

[0065] The compound with anti-inflammatory effect in the present invention is a limonoid compound, and the structural formula of the compound is as follows:

[0066]

[0067] Compound 1: Phellodenin A

[0068]

[0069] Compound 2: Phellodenin B

[0070]

[0071] Compound 3: Phellodenin C

[0072]

[0073] Compound 4: Phellodenin D

[0074]

[0075] Compound 5: Phellodenin E

[0076]

[0077] Compound 6: Phellodenin F

[0078]

[0079] Compound 7: Phellodenin G

[0080]

[0081] Compound 8: Phellodenin H

[0082]

[0083] Compound 9: 7α-limonol

[0084] Compound 10: 7β-limonol

[0085]

[0086] Compound 11: rutaevinexic acid

[0087]

[0088] Compound 12: limonexin

[0089]

[0090] Compound 13: isoobacunoic acid Compound 14: methyl isoobacunoate Compound 15: epi-isoobacunoic acid

[0091]

[0092] Compound 16: limonin

[0093] Compound 17: 12α-hydroxylimonin

[0094]

[0095] Compound 18: jangomolide

[0096]

[0097] Compound 19: obacunone

[0098]

[0099] Compound 20: kihadanin A

[0100]

[0101] Compound 21: kihadanin B

[0102]

[0103] Compound 22: kihadanin C

[0104]

[0105] Compound 23: dictangustone A

[0106] The compounds of the present invention were isolated and purified from the medicinal plant Phellodendron chinense Schneid. (Rutaceae, Phellodendron). The present invention obtained the above-mentioned limonoid compounds 1-23, which were discovered and prepared from Phellodendron chinense Schneid. from Enshi, by repeatedly subjecting the chloroform and ethyl acetate extracts of the 95% ethanol extract of Phellodendron chinense Schneid. from Enshi to column chromatography for separation and purification. Compound 1 is 1,4:14,15-diepoxy-7,8-seco-3,19:16,17-dilactone limonin. Compounds 2-3, 9-12, 16-17 are 1,4:14,15-diepoxy-3,19:16,17-dilactone limonins. Compound 4 is 1,4:14,15-diepoxy-3,19-lactone limonin. Compounds 5, 13-15 are 1,4:14,15-diepoxy-16,17-lactone limonins. Compounds 6, 19-22 are 14,15-epoxy-3,4:16,17-dilactone limonins. Compounds 7-8, 23 are 14,15-epoxy-16,17-lactone limonins. Compound 18 is 4,19-epoxy-3,19:16,17-dilactone limonin.

[0107] The structures were determined by using a variety of spectroscopic analysis methods, quantum chemical calculation methods, and X-ray single crystal diffraction methods.

[0108] Compounds 1-23 provided by the present invention can all inhibit inflammation and can be used to prepare anti-inflammatory drugs, specifically for preparing drugs for treating inflammations such as ulcerative colitis, pneumonia, hepatitis, etc.

[0109] Another embodiment of the present invention constructed an acute ulcerative colitis mouse model by the DSS (dextran sulfate sodium) induction method, especially for the in vivo anti-inflammatory activities of compounds 9 and 13. The results showed that they had significant protective effects, and the protective effects were stronger than those of the positive drug 5-aminosalicylic acid. The results of pathological sections of colon tissues showed that most of the mucosal epithelium in the colon sections proliferated and repaired, the infiltration of inflammatory cells was significantly reduced, the number of glandular cells and goblet cells increased, and the arrangement was relatively regular. The degree of protection of the glandular structure was better than that of the positive drug 5-aminosalicylic acid.

[0110] The following are specific examples

[0111] Example 1

[0112] (1) Isolation and preparation of compounds 1-23 shown in Formula I

[0113] 1. Plant materials

[0114] 24.8 Kg of the bark of the medicinal plant Phellodendron chinense Schneid. (Phellodendron amurense Rupr. var. sutchuenense Schneid.) of the genus Phellodendron in the Rutaceae family was collected from Jianshi County, Enshi Tujia and Miao Autonomous Prefecture, Hubei Province (latitude 30°30′, longitude 109°72′) in May 2021.

[0115] 2. Isolation and preparation process

[0116] After the dry Enshi Phellodendron chinense Schneid. (24.8 kg) was ground into coarse powder by a pulverizer, it was soaked and extracted with 95% ethanol. The extract was concentrated under reduced pressure to recover ethanol. After combining and evaporating the solvent to dryness, the total extract of Phellodendron chinense Schneid. (4.2 kg) was obtained. The total extract was suspended in an appropriate amount of warm distilled water and dispersed in a 25 L Longkou bottle. Then, it was successively extracted multiple times with organic solvents of different polarities (chloroform and ethyl acetate) until the color of the organic phase was relatively light. The solvent was recovered by concentration under reduced pressure, and the chloroform fraction (360.2 g) and ethyl acetate fraction (52.6 g) of the ethanol extract of Phellodendron chinense Schneid. were obtained respectively. After the chloroform fraction was dissolved with a dichloromethane / methanol mixed solvent, it was mixed with 1.5 times the mass of silica gel of 100-200 mesh. 10 times the amount of silica gel of 100-200 mesh was packed into a column, and gradient elution was carried out with a dichloromethane / methanol system, and its concentration ratio was 100:1 to 1:1. Combined with normal-phase thin-layer chromatography TLC development, it was divided into 12 components A1-A12. After the ethyl acetate fraction was dissolved with a dichloromethane / methanol mixed solvent, it was mixed with 1.5 times the mass of silica gel of 100-200 mesh. 10 times the amount of silica gel of 100-200 mesh was packed into a column, and gradient elution was carried out with a dichloromethane / methanol system, and its concentration ratio was 80:1 to 1:1. Combined with normal-phase thin-layer chromatography TLC development, it was divided into 8 components B1-B8.

[0117] The A3 fraction was separated by gradient elution on an ODS reverse-phase column chromatography (methanol-water, 30:70 → 100:0) to obtain 17 sub-fractions A3a - A3q. The fraction A3l was separated by gel column chromatography into 5 sub-fractions A3l1 - A3l5, among which A3l2 was further purified by semi-preparative HPLC (C18, methanol-water, 70:30, flow rate 1.5 mL / min) to obtain compound 19 (9.8 mg, tR = 17.0 min). The A4 fraction was separated by gradient elution on an ODS reverse-phase column chromatography (methanol-water, 30:70 → 100:0) to obtain 16 sub-fractions A4a - A4p. The fraction A4f was separated by gel column into 3 sub-fractions A4f1 - A4f3, and the fraction A4f3 was separated by normal-phase column chromatography to obtain 4 sub-fractions A4f3a - A4fd. The sub-fraction A4fd was further purified by semi-preparative HPLC (C18, methanol-water-formic acid, 54:46:0.1, flow rate 1.5 mL / min) to obtain compound 21 (11.0 mg, tR = 29.8 min) and 20 (13.1 mg, tR = 32.0 min). The fraction A4h was separated by gel column chromatography into 3 sub-fractions A4h1 - A4h, among which the fraction A4h3 was recrystallized to obtain compound 16 (50.0 mg). The fraction A4i was separated by gel column chromatography into 3 sub-fractions A4i1 - A4i3, among which the fraction A4i3 was purified by semi-preparative HPLC (C18, methanol-water, 70:30, flow rate 1.5 mL / min) to obtain compound 18 (9.3 mg, tR = 17.3 min). The A5 fraction was separated by gradient elution on an ODS reverse-phase column chromatography (methanol-water, 20:80 → 100:0) to obtain 14 sub-fractions A5a - A5n. The fraction A5h was separated by normal-phase silica gel column to obtain 5 sub-fractions A5h1 - A5h5, among which the fraction A5h3 was recrystallized to obtain compound 17 (308.5 mg). The fraction A5j was separated by normal-phase silica gel column to obtain 10 sub-fractions A5j1 - A5j10. Among them, the fraction A5j5 was separated by gel column chromatography into 3 sub-fractions A5j5a - A5j5c. The fraction A5j5a was recrystallized to obtain compound 9 (13.4 mg), and A5j5c was separated by semi-preparative HPLC (C18, methanol-water, 62:38, flow rate 1.5 mL / min) to obtain compound 10 (11.8 mg, tR = 14.5 min). The fraction A5j7 was separated by gel column chromatography to obtain compound 22 (7.3 mg). The A7 fraction was separated by normal-phase silica gel column chromatography combined with TLC color development to obtain 9 sub-fractions A7a - A7i. The fraction A7e was separated by gel column chromatography into 3 sub-fractions A7e1 - A7e3, and the sub-fraction A7e2 was further separated by normal-phase silica gel column into 5 sub-components A7e2a - A7e2e, among which the fraction A7e2e was recrystallized again to obtain compound 3 (9.0 mg).A7f was separated into 5 subcomponents A7f1 - A7f5 by normal-phase silica gel column. A7f3 was separated into 6 subcomponents A7f3a - A7f3f by reversed-phase medium-pressure column (C18 column, methanol - water, 30:70 → 65:35). Component A7f3f was purified by semi-preparative HPLC (C18, acetonitrile - water - formic acid, 42:58:0.1, flow rate 1.5 mL / min) to obtain compound 11 (6.7 mg, tR = 20.0 min) and 12 (6.1 mg, tR = 15.3 min). Component A9 was separated into 19 subcomponents A9a - A9s by reversed-phase medium-pressure column chromatography (methanol - water, 20:80 → 100:0) combined with TLC color development. Component A9j was separated into 3 subcomponents A9j1 - A9j3 by gel column chromatography. Among them, subcomponent A9j2 was purified by normal-phase silica gel column to obtain compound 13 (25.0 mg). Component A9m was separated into 4 subcomponents A9e1 - A9e4 by gel column chromatography. After combining A9e2 - A9e3, the main components were purified by normal-phase silica gel column (dichloromethane and methanol solvent system) and gel column respectively, and then further purified by semi-preparative HPLC (C18, methanol - water, 73:27, flow rate 1.5 mL / min) to obtain compound 6 (1.6 mg, tR = 23.0 min). Components A10 - A11 were combined and dissolved in pure methanol, and the chlorophyll components were removed by MCI column. Then, gradient elution was performed using an ODS reversed-phase column chromatography (methanol - water, 20:80 → 100:0) to obtain 17 subcomponents A10a - A10q. Component A10i was separated into 2 subcomponents A10i1 - A10i2 by gel column chromatography. Among them, component A10i1 was further purified by normal-phase silica gel column and semi-preparative HPLC (phenyl column, methanol - water, 70:30, flow rate 1.5 mL / min) to obtain monomer compound 1 (2.8 mg, tR = 27.4 min).

[0118] Component B4 was separated into 12 sub-components B4a - B4l by ODS reversed-phase column chromatography (methanol - water, 10:90 → 100:0) in combination with the color development of thin-layer chromatography. Component B4h was first purified by gel column to obtain the main component, and then further purified by HPLC (phenyl column, methanol - water, 80:20, flow rate 1.5 mL / min) to obtain monomer compound 5 (3.3 mg, tR = 25.5 min). Component B4i was purified by gel column chromatography (methanol) and semi-preparative HPLC (phenyl column, methanol - water, 78:22, flow rate 1.5 mL / min) to obtain compound 8 (4.4 mg, tR = 23.6 min). Component B4l was separated by gel column chromatography (methanol) into 4 sub-components B4l1 - B4l4. According to the color development results of thin-layer chromatography, component B4l2 was separated by normal-phase silica gel column chromatography into 7 sub-components B4l2a - B4l2g. Among them, B4l2c was purified by semi-preparative HPLC (C18, methanol - water, 74:36, flow rate 1.5 mL / min) to obtain monomer compounds 14 (12.3 mg, tR = 20.5 min) and 15 (7.2 mg, tR = 30.4 min). Component B5 was separated into 9 sub-components B5a - B5i by ODS reversed-phase column chromatography (methanol - water, 10:90 → 100:0). Component B5d was divided into 2 sub-components B5d1 - B5d2 by gel column (methanol). Component B5d2 was separated into 9 sub-components B5d2a - B5d2g by normal-phase silica gel column in combination with the color development of thin-layer chromatography. Component B5d2f was further purified by semi-preparative HPLC (C18 column, methanol - water, 51:49, flow rate 1.5 mL / min) to obtain compound 2 (11.0 mg, tR = 21.3 min). Component B6 was separated into 7 sub-components B6a - B6g by gradient elution of ODS reversed-phase column chromatography (methanol - water, 20:80 → 80:20). Component B6d was separated by gel column into 2 sub-components B6d1 - B6d2. Component B6d2 was separated by medium-pressure reversed-phase column (C18, methanol - water, 30:70 → 100:0) into 6 sub-components B6d2a - B6d2f. Among them, component B6d2d was purified by semi-preparative HPLC (C18, methanol - water - formic acid, 70:30:0.1, flow rate 1.5 mL / min) to obtain compound 23 (6.7 mg, tR = 25.6 min) and 7 (7.0 mg, tR = 29.9 min). Component B7 was separated into 9 sub-components B7a - B7i by ODS reversed-phase column chromatography (methanol - water, 0:100 → 80:20).Component B7e was separated into 7 sub-components B7e1 - B7e7 by normal-phase column chromatography. The main component of component B7e6 was repeatedly purified by gel filtration and then monomeric compound 4 (4.3 mg, tR = 18.5 min) was obtained by HPLC (C18, methanol - water, 20:80, flow rate 1.5 mL / min).

[0119] 3. Structure identification of compounds 1 - 23 shown in Formula I

[0120] Combined with a variety of spectroscopic analysis methods (high-resolution mass spectrometry HRESIMS, ultraviolet spectroscopy UV, infrared spectroscopy IR, and nuclear magnetic resonance spectroscopy NMR), quantum chemical calculation methods ( 13 C-NMR DP4+ probability analysis and ECD), and X-ray single crystal diffraction and other methods for comprehensive analysis to determine the structures of new compounds 1 - 8. The structures of known compounds 9 - 23 were determined by comparing literature data. Among them, the absolute configurations of compounds 2, 6, 9, 12, 17, 18, 19 were determined by X-ray single crystal diffraction, as shown in Figure 2 、 Figure 6 、 Figures 9 - 13 .

[0121] Compound 1: White powder; UV (methanol) λ max (logε): 212(0.72); IR (KBr) v max : 3414, 2976, 2944, 1740, 1630, 1383, 1286, 1191, 1131, 1030 cm -1 ; CD (methanol) λ(Δε): 214(-2.4), 263(+0.2) and 299(-0.1) nm; 1 1H and 13 13C NMR data are shown in Table 1 and Table 2; [α] 25 D -46.1 (c 0.1, methanol); high-resolution mass spectrometry m / z 527.1881 [M+Na] + (C 26 24 32 H 10 12 + (C 52 48 64 H 20 [[ID=?]]O Na's calculated value is 1031.3889).

[0122] By comparing the calculated and experimental ECD spectra, the absolute configuration of compound 1 was determined to be 1S,5R,8R,9R,10R,^{13}S,14S,15S,^{17}S. The calculated spectral band at 214 nm corresponds to the measured ECD spectrum, as shown inFigure 1 as shown

[0123] Compound 2: needle crystals (methanol: water = 5:1); melting point 301.2 - 303.3 °C; UV (methanol) λ max (logε): 214 (0.72); IR (KBr) ν max : 3398, 2957, 2925, 2856, 1736, 1601, 1413, 1384, 1285, 1025, 876, 807 cm -1 ; CD (methanol) λ(Δε): 203 (-0.8) and 235 (-2.9) nm; 1 H and 13 C NMR data are shown in Tables 1 and 2; [α] 25 D -22.5 (c 0.1, methanol); high resolution mass spectrometry m / z 511.1940 [M+Na] + (C 26 H 32 O9Na calculated value is 511.1944), 999.3977 [2M+Na] + (C 52 H 64 O 18 Na calculated value is 999.3990).

[0124] By comparing the calculated and experimental ECD spectra, the absolute configuration of Compound 2 was determined to be 1S,5R,7R,8S,9R,10R,12S,13S,14S,15S,17S. The calculated spectral band at 235 nm corresponded to the measured ECD spectrum, and the structure and absolute configuration of Compound 2 were confirmed by single crystal X-ray diffraction experiment induced by copper target, with the Flack parameter being 0.06 (12), as Figure 2 shown

[0125] Compound 3: white powder; UV (methanol) λ max (logε): 212 (0.74); IR (KBr) ν max : 3437, 2972, 2939, 2306, 1746, 1624, 1373, 1283, 1053, 1028, 945, 815, 536 cm -1 ; CD (methanol) λ(Δε): 208 (+1.0), 234 (-2.2) and 294 (-1.4) nm; 1 H and 13 C NMR data are shown in Tables 1 and 2; [α] 26 D-76.8 (c 0.1, methanol); HRMS m / z 541.1678 [M+Na] (calcd for C H O Na 541.1686). + (C 26 H 30 O 11 The absolute configuration of compound 3 was determined to be 1S,5R,8R,9R,10R,12S,13S,14R,15S,17R by comparing the calculated and experimental ECD spectra. The calculated bands at 208, 234 and 294 nm corresponded to the experimental ECD spectrum as shown below.

[0126] Figure 3 as

[0127] Compound 4: White powder; UV (methanol) λ (logε): 212 (0.70), 280 (0.21); IR (KBr) ν: 3406, 2953, 1751, 1709, 1454, 1383, 1297, 1235, 1155, 1025, 847, 834, 601 cm max (logε): 212 (0.70), 280 (0.21); IR (KBr) ν max : 3406, 2953, 1751, 1709, 1454, 1383, 1297, 1235, 1155, 1025, 847, 834, 601 cm -1 ; CD (methanol) λ (Δε): 206 (-3.7), 243 (+0.4) and 310 (-1.7) nm; 1 H and 13 C NMR data are shown in Tables 1 and 2; [α] 25 D -78.1 (c 0.1, methanol); HRMS m / z 523.1934 [M+Na] (calcd for C H O Na 523.1944), 1023.4010 [2M+Na] (calcd for C H O Na 1023.3990). + (C 27 H 32 O9Na (calcd for C H O Na 523.1944), 1023.4010 [2M+Na] (calcd for C H O Na 1023.3990). + (C 54 H 64 O 18 [[ID= / / ID=43]]The absolute configuration of compound 4 was determined to be 1S,5R,8R,9R,10R,13R,14R,15S by comparing the calculated and experimental ECD spectra. The calculated bands at 206, 243 and 310 nm corresponded to the experimental ECD spectrum as shown below.

[0128] Figure 4 as

[0129] Compound 5: White powder; UV (methanol) λ max (logε): 212 (0.86), 280 (0.29); IR (KBr) ν max ​​: 3420, 2959, 2932, 1739, 1605, 1457, 1384, 1270, 1127, 1028 cm -1 ; CD( methanol) λ(Δε): 210(+1.0), 234(-1.5) and 294(-1.0) nm; 1 H and 13 C NMR data are shown in Tables 3 and 4; [α] 26 D -56.7 (c 0.1, methanol); high resolution mass spectrometry m / z 567.2561 [M+Na] + (C 30 H 40 O9Na is calculated to be 567.2570), 1111.5197 [2M+Na] + (C 60 H 80 O 18 Na is calculated to be 1111.5242).

[0130] By comparing the calculated and experimental ECD spectra of Compound 5, the absolute configuration of Compound 5 was determined to be 1S,5R,8R,9R,10R,13S,14R,15S,17S. The calculated spectral bands at 210, 234 and 294 nm corresponded to the measured ECD spectrum, as Figure 5 shown.

[0131] Compound 6: Colorless needle crystals (methanol: water = 10:1); melting point 310.6 - 312.5 °C; UV (methanol) λ max (logε): 214(0.86),; IR (potassium bromide) v max : 3410, 2943, 1722, 1673, 1232, 1605, 1384, 1354, 1279, 1126, 1028, 809, 601 cm -1 ; CD (methanol) λ(Δε): 229(-2.6) and 255(+3.2) nm; 1 H and 13 C NMR data are shown in Tables 3 and 4; [α] 26 D -72.8 (c 0.1, methanol); high resolution mass spectrometry m / z 495.2003 [M+Na] + (C 26 H 32 O8Na is calculated to be 495.1995), 967.4084 [2M+Na] + (C 52 H 64 O 16The calculated value of Na is 967.4092), 1439.6081 [3M+Na] + (C 78 H 96 O 24 The calculated value of Na is 1439.6081).

[0132] By comparing the calculated and experimental ECD spectra of Compound 6, the absolute configuration of Compound 6 was determined to be 5S, 6S, 7S, 8S, 9R, 10R, 13S, 14R, 15S, 17S. The calculated spectral bands at 229 and 255 nm corresponded to the measured ECD spectrum. And through the single-crystal X-ray diffraction experiment induced by a copper target, the Flack parameter was 0.01(14), verifying the structure and absolute configuration of Compound 6, as Figure 6 shown.

[0133] Compound 7: White powder; UV (methanol) λ max (logε): 214(0.81); IR (KBr) v max : 3408, 2979, 1712, 1630, 1586, 1445, 1390, 1358, 1292, 1165, 1024, 1004, 837, 601 cm -1 ; CD (methanol) λ(Δε): 204(+0.3), 215(-0.8), 223(-0.2), 240(-1.7), 262(-0.3) and 296(-1.7) nm; 1 H and 13 C NMR data are shown in Tables 3 and 4; [α] 25 D -14.5 (c 0.1, methanol); High-resolution mass spectrometry m / z 495.1982 [M+Na] + (C 26 H 32 O8Na's calculated value is 495.1995), 967.3962 [2M+Na] + (C 52 H 64 O 16 The calculated value of Na is 967.4092) as Figure 7 shown.

[0134] Due to the lack of key signals in NMR, the connection of the A / B ring could not be proven, so computational chemistry was used for its analysis: A conformational search was performed on Compound 7 to obtain the conformational molecules arranged from largest to smallest in Boltzmann distribution and their distribution probabilities. The free energy barriers of the four conformations with the highest distribution probabilities were selected, including zero-point and thermal correction values as well as the solvent effect of methanol, and calculated to be 16.4 kcal mol -1Using the B3LYP / 6-31G DFT method, the TMS-corrected 13 C chemical shifts of A-1 - A-4 were calculated using GIAO (Table 5), and the reasons for their 13 unidentifiability in the 13 C NMR spectrum were obtained: the chemical shift difference exceeded 2 ppm. The greater the chemical shift difference, the greater the deviation between the conformations, and the broader or weaker the carbon resonances in the

[0135] C NMR spectrum. Finally, the absolute configuration was determined to be 5R,8R,9R,10R,13S,14R,15S,17S by ECD.

[0135] Compound 8: White powder; UV (methanol) λ max (logε): 214 (0.87); IR (KBr) ν max : 3411, 2954, 1742, 1713, 1613, 1440, 1384, 1279, 1201, 1169, 1025, 934, 824, 687 cm -1 ; CD (methanol) λ (Δε): 222 (+0.3), 244 (-2.5), 262 (-0.7) and 295 (-2.8) nm; 1 H and 13 C NMR data are shown in Tables 3 and 4; [α] 26 D -11.7 (c 0.1, methanol); HRMS m / z 509.2140 [M+Na] + (C 27 H 34 O8Na calcd. 509.2151), 995.4400 [2M+Na] + (C 54 H 68 O 16 Na calcd. 995.4405), 1481.6642 [3M+Na] + (C 81 H 102 O 24 Na calcd. 1481.6659).

[0136] By comparing the calculated and experimental ECD spectra of Compound 8, the absolute configuration of Compound 8 was determined to be 5R,8R,9R,10R,13S,14R,15S,17S. The calculated spectral bands at 244, 262, and 295 nm corresponded to the measured ECD spectrum, as Figure 8 shown.

[0137] Table 1 1 H NMR data of Compounds 1 - 4 (deuterochloroform, 400 MHz)

[0138]

[0139]

[0140] Table 2 of Compounds 1 - 4 13 $^{13}$C NMR Data (CD$_3$OD, 100 MHz)

[0141]

[0142]

[0143] Table 3 of Compounds 5 - 8 1 $^1$H NMR Data (CD$_3$OD, 400 MHz)

[0144]

[0145] Table 4 of Compounds 5 - 8 13 $^{13}$C NMR Data (CD$_3$OD, 100 MHz)

[0146]

[0147]

[0148] Table 5 of TMS - corrected $^{13}$C Chemical Shifts of the Major Conformations A - 1 - A - 4 of Compound 7 13 $^{13}$C Chemical Shifts

[0149]

[0150] Compound 9: [α] 26 D - 57.3 (c 0.1, methanol); white powder; 1 $^1$H NMR (CDCl$_3$, 400 MHz): δ H7.38 (2H, br s, H-21 and H-23), 6.31 (1H, s, H-22), 5.55 (1H, s, H-17), 4.46 (1H, d, J = 13.1 Hz, H-19β), 4.38 (1H, d, J = 13.1 Hz, H-19α), 4.01 (1H, d, J = 3.3 Hz, H-7), 3.85 (1H, s, H-15), 3.56 (1H, br s, H-1), 2.90 (1H, dd, J = 16.6, 3.8 Hz, H-6α), 2.72 (1H, dd, J = 12.8, 6.6 Hz, H-2α), 2.60 (1H, overlap, H-2β), 2.57 (1H, overlap, H-6β), 1.27 (3H, s, H3-18), 1.25 (3H, s, H3-28), 1.11 (3H, s, H3-29), 0.85 (3H, s, H3-30); 13 C NMR (chloroform-d, 100 MHz): δ C 80.1 (C-1), 36.0 (C-2), 170.2 (C-3), 81.0 (C-4), 52.1 (C-5), 27.6 (C-6), 70.4 (C-7), 44.0 (C-8), 42.0 (C-9), 45.9 (C-10), 17.7 (C-11), 26.3 (C-12), 38.6 (C-13), 69.4 (C-14), 57.6 (C-15), 168.3 (C-16), 78.6 (C-17), 18.9 (C-18), 66.2 (C-19), 120.6 (C-20), 141.5 (C-21), 110.1 (C-22), 143.3 (C-23), 17.7 (C-30), 21.8 (C-29), 30.5 (C-28).

[0151] Compound 10: [α] 26 D +7.2 (c 0.1, methanol); white powder; 1 H NMR (methanol-d4, 400 MHz): δ H7.51(1H,br s,H-21), 7.50(1H,dd,J = 1.9,1.4Hz,H-23), 6.42(1H,br s,H-22), 5.63(1H,s,H-17), 4.62(1H,d,J = 13.1Hz,H-19β), 4.59(1H,d,J = 13.1Hz,H-19α), 4.49(1H,s,H-15), 4.09(1H,dd,J = 3.6,1.8Hz,H-1), 3.82(1H,dd,J = 10.3,5.0Hz,H-7), 2.79(1H,dd,J = 16.5,1.8Hz,H-2α), 2.73(1H,dd,J = 16.5,3.6Hz,H-2β), 2.04(1H,overlap,H-6α), 1.80(1H,overlap,H-6β), 1.27(3H,s,H3-18), 1.26(3H,s,H3-28), 1.10(3H,s,H3-29), 0.97(3H,s,H3-30); 13 C NMR(chloroform-d, 100 MHz): δ C 81.0(C-1), 36.9(C-2), 173.8(C-3), 82.1(C-4), 59.1(C-5), 29.3(C-6), 78.8(C-7), 45.2(C-8), 47.1(C-9), 47.2(C-10), 19.0(C-11), 27.3(C-12), 40.6(C-13), 74.4(C-14), 57.0(C-15), 170.8(C-16), 79.8(C-17), 19.1(C-18), 67.5(C-19), 122.2(C-20), 143.0(C-21), 111.2(C-22), 144.6(C-23), 14.3(C-30), 22.0(C-29), 30.9(C-28).

[0152] Compound 11: [α] 24 D -68.3(c 0.1, methanol); white powder; 1 H NMR(pyridine-d5, 400 MHz): δ H7.78 / 7.75(1H,br s,H-22), 6.70 / 6.60(1H,br s,H-23), 5.78(1H,s,H-17), 5.19(1H,d,J = 13.1Hz,H-19β), 4.70(1H,d,J = 13.1Hz,H-19α), 4.99(1H,s,H-15), 3.24(1H,t,J = 2.8Hz,H-6β), 3.05(1H,d,J = 16.2Hz,H-2α), 2.65(1H,d,J = 16.2Hz,H-2β), 2.55(1H,d,J = 14.9Hz,H-6α), 1.38(3H,s,H3-18), 1.27(3H,s,H3-28), 1.26(3H,s,H3-29), 1.21(3H,s,H3-30); 13 C NMR(deuterated pyridine, 100 MHz): δ C 80.4(C-1), 37.3(C-2), 170.5(C-3), 80.9(C-4), 58.2(C-5), 36.9(C-6), 208.2(C-7), 50.6(C-8), 47.1(C-9), 46.7(C-10), 18.9(C-11), 28.6(C-12), 39.9(C-13), 66.1(C-14), 53.0(C-15), 166.9(C-16), 77.7(C-17), 18.5(C-18), 66.0(C-19), 167.5(C-20), 98.1(C-21), 121.9(C-22), 169.0(C-23), 16.4(C-30), 22.2(C-29), 30.4(C-28).

[0153] Compound 12: [α] 26 D -42.6(c 0.1, methanol); white powder; 1 H NMR(deuterated chloroform, 400 MHz): δ H6.68 (1H, br s, H-21), 6.65 (1H, br s, H-22), 5.86 (1H, s, H-17), 5.28 (1H, d, J = 13.1 Hz, H-19β), 4.74 (1H, d, J = 13.1 Hz, H-19α), 4.52 (1H, s, H-15), 4.39 (1H, d, J = 3.7 Hz, H-1), 3.33 (1H, dd, J = 15.7, 14.3 Hz, H-6β), 3.22 (1H, dd, J = 16.5, 3.7 Hz, H-2α), 3.12 (1H, d, J = 16.4 Hz, H-2β), 2.88 (1H, dd, J = 11.6, 3.8 Hz, H-9), 2.70 (1H, dd, J = 15.7, 3.4 Hz, H-6α), 2.59 (1H, dd, J = 14.3, 3.3 Hz, H-5), 1.43 (3H, s, H3-18), 1.28 (3H, s, H3-28), 1.28 (3H, s, H3-29), 1.22 (3H, s, H3-30); 13 13C NMR (chloroform-d, 100 MHz): δ C 80.3 (C-1), 37.4 (C-2), 170.6 (C-3), 80.9 (C-4), 60.9 (C-5), 37.0 (C-6), 208.2 (C-7), 52.5 (C-8), 48.8 (C-9), 46.9 (C-10), 19.4 (C-11), 30.9 (C-12), 39.2 (C-13), 67.0 (C-14), 54.6 (C-15), 166.9 (C-16), 79.3 (C-17), 21.2 (C-18), 66.4 (C-19), 164.0 (C-20), 99.5 (C-21), 123.5 (C-22), 162.0 (C-23),

[0154] 17.5 (C-30), 22.2 (C-29), 30.3 (C-28).

[0155] Compound 13: [α] 26 D -11.3 (c 0.1, methanol); white powder; 1 1H NMR (chloroform-d, 400 MHz): δ H7.38(1H,br s,H-21), 7.36(1H,dd,J = 1.8,1.6Hz,H-23), 6.31(1H,d,J = 1.8Hz,H-22), 5.47(1H,s,H-17), 4.18(1H,dd,J = 11.4,3.1Hz,H-1), 4.12(1H,s,H-15), 2.78(1H,dd,J = 15.4,14.2Hz,H-6β), 2.40(1H,dd,J = 14.2,11.7Hz,H-2α), 2.33(1H,dd,J = 14.2,3.5Hz,H-6α), 2.23(1H,dd,J = 14.2,3.1Hz,H-2β), 2.23(1H,dd,J = 14.2,3.1Hz,H-9), 2.08(1H,dd,J = 15.4,3.5Hz,H-5), 1.38(3H,s,H3-19), 1.30(3H,s,H3-28), 1.17(3H,s,H3-29), 1.18(3H,s,H3-30), 1.15(3H,s,H3-18); 13 C NMR(chloroform-d, 100 MHz): δ C 83.8(C-1), 36.9(C-2),

[0156] 174.7(C-3), 81.6(C-4), 56.1(C-5), 36.8(C-6), 208.0(C-7), 51.6(C-8), 43.6(C-9), 47.4(C-10), 18.4(C-11), 30.7(C-12), 38.5(C-13), 66.7(C-14), 54.1(C-15), 167.3(C-16), 78.1(C-17), 21.2(C-18), 19.9(C-19), 120.4(C-20), 141.3(C-21), 110.0(C-22),

[0157] 143.4(C-23), 18.0(C-30), 23.9(C-29), 30.7(C-28).

[0158] Compound 14: [α] 26 D -21.3(c 0.1, methanol); white powder; 1 H NMR(chloroform-d, 400 MHz): δ H7.38(1H,br s,H-21), 7.37(1H,dd,J = 1.8,1.6Hz,H-23), 6.32(1H,d,J = 1.8Hz,H-22), 5.47(1H,s,H-17), 4.20(1H,dd,J = 11.1,3.4Hz,H-1), 4.11(1H,s,H-15), 3.67(3H,s,OCH3), 2.79(1H,dd,J = 15.2,14.6Hz,H-6β), 2.39(1H,dd,J = 13.4,11.1Hz,H-2α), 2.32(1H,dd,J = 13.9,3.4Hz,H-6α), 2.23(1H,dd,J = 11.3,2.9Hz,H-2β), 2.20(1H,dd,J = 13.8,3.3Hz,H-9), 2.06(1H,dd,J = 15.2,3.5Hz,H-5), 1.37(3H,s,H3-19), 1.26(3H,s,H3-28), 1.17(3H,s,H3-29), 1.15(3H,s,H3-30), 1.14(3H,s,H3-18); 13 13C NMR(chloroform-d, 100 MHz): δ C 84.0(C-1), 37.2(C-2),

[0159] 171.4(C-3), 80.9(C-4), 56.2(C-5), 36.9(C-6), 208.3(C-7), 51.6(C-8), 43.7(C-9), 47.3(C-10), 18.4(C-11), 30.7(C-12), 38.5(C-13), 66.7(C-14), 54.4(C-15), 167.3(C-16), 78.1(C-17), 21.2(C-18), 19.9(C-19), 120.4(C-20), 141.3(C-21), 110.0(C-22), 1,43.4(C-23), 18.0(C-30), 23.9(C-29), 30.7(C-28), 52.2(OCH3).

[0160] Compound 15: [α] 26 D +9.8(c 0.1, methanol); white powder; 1 1H NMR(methanol-d, 400 MHz): δ H7.52(1H,br s,H-21), 7.49(1H,dd,J = 1.8,1.6Hz,H-23), 6.43(1H,d,J = 1.8Hz,H-22), 5.59(1H,s,H-17), 4.31(1H,s,H-15), 3.96(1H,dd,J = 8.7,2.7Hz,H-1), 2.90(1H,dd,J = 15.3,14.6Hz,H-6β), 2.60(1H,dd,J = 15.8,2.7Hz,H-2α), 2.47(1H,dd,J = 12.0,4.2Hz,H-9), 2.38(1H,dd,J = 15.8,8.7Hz,H-2β), 2.26(1H,dd,J = 14.9,3.5Hz,H-6α), 2.14(1H,dd,J = 15.7,3.5Hz,H-5), 1.28(3H,s,H3-18), 1.22(3H,s,H3-28), 1.21(3H,s,H3-30), 1.19(3H,s,H3-19), 1.13(3H,s,H3-29); 13 C NMR(deuterated methanol, 100MHz): δ C 84.9(C-1), 38.3(C-2), 176.8(C-3), 79.8(C-4), 61.3(C-5), 37.6(C-6), 210.8(C-7), 52.4(C-8), 47.5(C-9), 47.2(C-10), 18.4(C-11), 30.7(C-12), 40.3(C-13), 69.4(C-14), 56.4(C-15), 170.3(C-16), 79.8(C-17), 19.4(C-18), 13.0(C-19), 122.1(C-20), 142.9(C-21), 111.2(C-22), 144.6(C-23), 20.6(C-30), 23.9(C-29), 30.2(C-28).

[0161] Compound 16: [α] 26 D -12.7(c 0.1, methanol); white powder; 1 H NMR(deuterated chloroform, 400MHz): δ H7.39 (1H, br s, H-21), 7.38 (1H, dd, J = 1.8, 0.9 Hz, H-23), 6.32 (1H, d, J = 1.8 Hz, H-22), 5.45 (1H, s, H-17), 4.74 (1H, d, J = 13.1 Hz, H-19β), 4.44 (1H, d, J = 13.1 Hz, H-19α), 4.02 (1H, s, H-15), 4.02 (1H, br s, H-1), 2.96 (1H, dd, J = 16.8, 3.8 Hz, H-2β), 2.66 (1H, dd, J = 16.8, 2.0 Hz, H-2α), 2.53 (1H, dd, J = 12.3, 2.9 Hz, H-9), 2.83 (1H, dd, J = 15.9, 14.5 Hz, H-6β), 2.45 (1H, dd, J = 14.5, 3.3 Hz, H-6α), 2.20 (1H, dd, J = 15.9, 3.3 Hz, H-5), 1.27 (3H, s, H3-28), 1.162 (3H, s, H3-29), 1.155 (3H, s, H3-18), 1.05 (3H, s, H3-30); 13 13C NMR (chloroform-d, 100 MHz): δ C 79.4 (C-1), 35.9 (C-2), 169.3 (C-3), 80.5 (C-4), 60.8 (C-5), 36.6 (C-6), 206.3 (C-7), 51.6 (C-8), 48.4 (C-9), 46.2 (C-10), 19.2 (C-11), 31.1 (C-12), 38.2 (C-13), 65.9 (C-14), 54.1 (C-15), 166.8 (C-16), 78.0 (C-17), 21.6 (C-18), 65.9 (C-19), 120.2 (C-20), 141.3 (C-21), 109.9 (C-22), 143.3 (C-23), 17.9 (C-30), 20.9 (C-29), 30.4 (C-28).

[0162] Compound 17: [α] 25 D 25 D -38.0 (c 0.1, methanol); white powder; 1 1H NMR (pyridine-d5, 400 MHz): δ H7.85 (1H, br s, H-21), 7.55 (1H, br s, H-23), 6.72 (1H, d, J = 1.8 Hz, H-22), 6.68 (1H, d, J = 3.96 Hz, OH-12), 5.87 (1H, s, H-17), 5.33 (1H, d, J = 13.0 Hz, H-19β), 4.67 (1H, d, J = 13.0 Hz, H-19α), 4.39 (1H, overlap, H-1), 4.39 (1H, overlap, H-12), 4.32 (1H, s, H-15), 3.37 (1H, dd, J = 15.2, 13.4 Hz, H-6β), 3.01 (2H, d, J = 2.8 Hz, H-11), 4.74 (1H, d, J = 13.1 Hz, H-19β), 4.44 (1H, d, J = 13.1 Hz, H-19α), 4.02 (1H, s, H-15), 4.02 (1H, br s, H-1), 2.96 (1H, dd, J = 16.8, 3.8 Hz, H-2β), 2.66 (1H, dd, J = 16.8, 2.0 Hz, H-2α), 2.53 (1H, dd, J = 12.3, 2.9 Hz, H-9), 2.83 (1H, dd, J = 15.9, 14.5 Hz, H-6β), 2.45 (1H, dd, J = 14.5, 3.3 Hz, H-6α), 2.20 (1H, dd, J = 15.9, 3.3 Hz, H-5), 1.27 (3H, s, H3-18), 1.162 (3H, s, H3-28), 1.155 (3H, s, H3-30), 1.05 (3H, s, H3-29); 13 13C NMR (pyridine-d5, 100 MHz): δ C 80.2 (C-1), 37.4 (C-2), 170.6 (C-3), 80.7 (C-4), 61.8 (C-5), 33.4 (C-6), 207.8 (C-7), 51.6 (C-8), 45.9 (C-9), 46.7 (C-10), 36.8 (C-11), 68.2 (C-12), 44.4 (C-13), 65.8 (C-14), 53.5 (C-15), 167.8 (C-16), 78.4 (C-17), 14.4 (C-18), 66.9 (C-19), 121.7 (C-20), 142.6 (C-21), 111.7 (C-22), 144.0 (C-23), 17.0 (C-30), 22.1 (C-29), 30.3 (C-28).

[0163] Compound 18: [α] 26 D+18.6 (c 0.1, methanol); white powder; 1 H NMR (chloroform-d, 400 MHz): δ H 7.39 (1H, d, J = 0.8 Hz, H-21), 7.38 (1H, dd, J = 1.8, 0.9 Hz, H-23), 6.51 (1H, d, J = 10.0 Hz, H-1), 6.30 (1H, dd, J = 1.8, 0.9 Hz, H-22), 6.11 (1H, d, J = 10.0 Hz, H-2), 6.04 (1H, s, H-19), 5.51 (1H, s, H-17), 3.97 (1H, s, H-15), 1.34 (3H, s, H3-28), 1.26 (3H, s, H3-29), 1.26 (3H, s, H3-30), 1.13 (3H, s, H3-18); 13 C NMR (chloroform-d, 100 MHz): δ C 150.9 (C-1), 120.1 (C-2), 160.5 (C-3), 86.6 (C-4), 53.4 (C-5), 37.8 (C-6), 209.2 (C-7), 49.3 (C-8), 42.0 (C-9), 49.6 (C-10), 19.3 (C-11), 29.3 (C-12), 38.6 (C-13), 67.5 (C-14), 54.4 (C-15), 166.9 (C-16), 78.0 (C-17), 20.4 (C-18), 104.2 (C-19), 120.1 (C-20), 141.4 (C-21), 109.9 (C-22), 143.5 (C-23), 15.9 (C-30), 25.1 (C-29), 29.3 (C-28).

[0164] Compound 19: [α] 26 D -11.0 (c 0.1, methanol); white powder; 1 H NMR (chloroform-d, 400 MHz): δ H7.40 (1H, m, H-21), 7.38 (1H, dd, J = 1.9, 1.8 Hz, H-23), 6.49 (1H, d, J = 11.7 Hz, H-1), 6.34 (1H, dd, J = 1.9, 0.9 Hz, H-22), 5.94 (1H, d, J = 11.7 Hz, H-2), 5.44 (1H, s, H-17), 3.63 (1H, s, H-15), 2.96 (1H, dd, J = 14.1, 14.0 Hz, H-6β), 2.57 (1H, dd, J = 14.1, 5.0 Hz, H-5), 2.27 (1H, dd, J = 14.0, 5.0 Hz, H-6α), 1.48 (3H, s, H3-29), 1.48 (3H, s, H3-19), 1.43 (3H, s, H3-28), 1.22 (3H, s, H3-18), 1.10 (3H, s, H3-30); 13 13C NMR (chloroform-d, 100 MHz): δ C 157.0 (C-1), 123.2 (C-2), 166.8 (C-3), 84.2 (C-4), 57.5 (C-5), 40.1 (C-6), 207.6 (C-7), 53.2 (C-8), 49.4 (C-9), 43.3 (C-10), 17.2 (C-11), 33.0 (C-12), 37.6 (C-13), 65.2 (C-14), 53.5 (C-15), 167.1 (C-16), 78.2 (C-17), 19.7 (C-18), 21.4 (C-19), 120.3 (C-20), 141.2 (C-21), 110.0 (C-22), 143.4 (C-23), 16.7 (C-30), 27.0 (C-29), 32.3 (C-28).

[0165] Compound 20: [α] 24 D -64.0 (c 0.1, methanol); white powder; 1 1H NMR (chloroform-d, 400 MHz): δ H6.59 (1H, d, J = 11.4, H-1), 6.24 (1H, br s, H-22), 6.04 / 6.01 (1H, br s, H-21), 5.95 (1H, d, J = 11.4 Hz, H-2), 5.32 (1H, br s, H-17), 3.56 (1H, s, H-15), 2.95 (1H, dd, J = 14.0, 13.9 Hz, H-6β), 2.62 (1H, dd, J = 13.9, 4.9 Hz, H-5), 2.27 (1H, dd, J = 14.0, 5.0 Hz, H-6α), 1.48 (3H, s, H3-29), 1.47 (3H, s, H3-19), 1.43 (3H, s, H3-28), 1.20 (3H, s, H3-18), 1.08 (3H, s, H3-30); 13 13C NMR (chloroform-d, 100 MHz): δ C 157.0 (C-1), 123.3 (C-2), 167.4 (C-3), 84.3 (C-4), 57.1 (C-5), 39.8 (C-6), 207.7 (C-7), 52.8 (C-8), 49.0 (C-9), 43.1 (C-10), 19.4 (C-11), 32.1 (C-12), 37.8 (C-13), 65.1 (C-14), 53.1 (C-15), 169.8 (C-16), 78.3 (C-17), 16.8 (C-18), 21.3 (C-19), 162.0 (C-20), 100.6 (C-21), 122.7 (C-22), 166.0 (C-23), 16.6 (C-30), 26.8 (C-29), 32.0 (C-28).

[0166] Compound 21: [α] 26 D -2.7 (c 0.1, methanol); white powder; 1 1H NMR (chloroform-d, 400 MHz): δ H7.34 / 7.31(1H,br s,H-22), 6.51 / 6.47(1H,d,J = 11.9,H-1), 6.20 / 6.21(1H,br s,H-21), 5.94 / 5.93(1H,d,J = 11.4Hz,H-2), 5.39 / 5.37(1H,br s,H-17), 3.67 / 3.66(1H,s,H-15), 2.95(1H,dd,J = 14.0,13.9Hz,H-6β), 2.58(1H,dd,J = 13.9,4.9Hz,H-5), 2.29(1H,dd,J = 14.0,5.0Hz,H-6α), 1.488(3H,s,H3-29), 1.483(3H,s,H3-19), 1.43(3H,s,H3-28), 1.23(3H,s,H3-18), 1.11 / 1.10(3H,s,H3-30); 13 C NMR(chloroform-d, 100 MHz): δ C 156.8(C-1), 123.0(C-2), 167.2(C-3), 84.2(C-4), 57.3(C-5), 39.8(C-6), 207.5(C-7), 52.7(C-8), 49.0(C-9), 43.1(C-10), 19.4(C-11), 32.2(C-12), 37.8(C-13), 65.0(C-14), 53.2(C-15), 169.3(C-16), 78.4(C-17), 16.8(C-18), 21.5(C-19), 162.0(C-20), 165.7(C-21), 123.2(C-22), 97.9(C-23), 16.6(C-30), 26.8(C-29), 32.0(C-28).

[0167] Compound 22: [α] 24 D -28.0(c 0.1, methanol); white powder; 1 H NMR(chloroform-d, 400 MHz): δ H6.58 (1H, d, J = 11.7 Hz, H-1), 5.97 (1H, d, J = 11.7 Hz, H-2), 4.93 (1H, d, J = 3.7 Hz, H-23), 4.90 (1H, s, H-17), 4.76 (1H, s, H-21), 4.13 (1H, dd, J = 10.4, 3.7 Hz, H-22), 3.49 (1H, s, H-15), 3.45 (1H, s, OCH3-23), 3.35 (1H, s, OCH3-21), 2.93 (1H, dd, J = 14.1, 14.1 Hz, H-6β), 2.81 (1H, d, J = 11.8 Hz), 2.57 (1H, dd, J = 14.1, 5.0 Hz, H-5), 2.24 (1H, dd, J = 14.0, 5.0 Hz, H-6α), 1.47 (3H, s, H3-29), 1.46 (3H, s, H3-19), 1.43 (3H, s, H3-28), 1.26 (3H, s, H3-18), 1.16 (3H, s, H3-30); 13 13C NMR (chloroform-d, 100 MHz): δ C 157.4 (C-1), 123.1 (C-2), 167.3 (C-3), 84.2 (C-4), 57.5 (C-5), 40.1 (C-6), 207.8 (C-7), 53.4 (C-8), 49.5 (C-9), 43.2 (C-10), 19.6 (C-11), 31.0 (C-12), 38.1 (C-13), 65.3 (C-14), 52.8 (C-15), 166.9 (C-16), 81.8 (C-17), 21.9 (C-18), 16.7 (C-19), 81.8 (C-20), 109.1 (C-21), 76.5 (C-22), 112.2 (C-23), 17.0 (C-30), 27.0 (C-29), 32.2 (C-28), 55.3 (OCH3-21), 56.7 (OCH3-23).

[0168] Compound 23: [α] 26 D -58.9 (c 0.1, methanol); white powder; 1 1H NMR (methanol-d, 400 MHz): δ H7.51 (1H, br s, H-21), 7.48 (1H, br s, H-23), 6.75 (1H, d, J = 16.0 Hz, H-1), 6.43 (1H, br s, H-22), 5.77 (1H, d, J = 16.0 Hz, H-2), 5.48 (1H, s, H-17), 3.68 (1H, s, H-15), 3.11 (1H, dd, J = 14.4, 13.8 Hz, H-6β), 2.53 (1H, dd, J = 14.4, 4.4 Hz, H-5), 2.16 (1H, dd, J = 13.8, 4.4 Hz, H-6α), 1.43 (3H, s, H3-19), 1.22 (3H, s, H3-29), 1.22 (3H, s, H3-28), 1.20 (3H, s, H3-30), 1.09 (3H, s, H3-18); 13 13C NMR (deuterated methanol, 100 MHz): δ C 161.0 (C-1), 121.9 (C-2), 169.7 (C-3), 74.3 (C-4), 61.0 (C-5), 40.1 (C-6), 212.4 (C-7), 54.4 (C-8), 49.4 (C-9), 45.7 (C-10), 17.2 (C-11), 33.0 (C-12), 37.6 (C-13), 66.9 (C-14), 54.5 (C-15), 169.7 (C-16), 80.0 (C-17), 19.5 (C-18), 16.2 (C-19), 123.2 (C-20), 142.7 (C-21), 111.1 (C-22), 144.6 (C-23), 30.0 (C-28), 30.8 (C-29), 21.6 (C-30).

[0169] Example 2

[0170] Compound 1-23 was applied to establish an in vitro inflammation model of lipopolysaccharide (LPS)-induced RAW264.7 cells. Monomeric compounds were used for intervention (dexamethasone as the positive drug), and the inflammatory regulation level of RAW264.7 cells was judged by measuring the release amount of NO in the cell supernatant. The results are shown in Table 6.

[0171] Table 6. Results of the inhibitory effect of Compound 1-23 on the release of NO from LPS-induced RAW264.7 cells a

[0172]

[0173] Note: a Mean ± standard deviation, n = 3; bThe positive drug is dexamethasone

[0174] Conclusion: All limonoid compounds (1 - 23) showed significant inhibitory effects on the release of NO from LPS - induced RAW264.7 cells, and the IC 50 values ranged from 4.2 ± 0.1 to 18.6 ± 5.0 μM. Among them, the IC 50 values of compounds 9 and 13 were 4.2 ± 0.1 and 4.8 ± 1.0 μM respectively, showing good anti - inflammatory effects.

[0175] Example 3

[0176] To further confirm the in - vivo anti - ulcerative colitis activity and mechanism of action of the representative limonoid compounds 7α - limonol (9) and isoobacunoic acid (13) with the best in - vitro anti - inflammatory activity and relatively high content, a mouse model of DSS - induced acute ulcerative colitis was established. The fur condition, activity sensitivity, mental state, and drinking water situation of the mice in the DSS - induced acute ulcerative colitis mouse model were observed. The body weight, diarrhea, and blood in the stool of the mice after modeling were recorded, and the disease activity index DAI score was calculated.

[0177] Conclusion: Compounds 9 and 13 significantly and dose - dependently reduced the DAI index of ulcerative colitis in DSS - modeled mice and increased the colon length of DSS - modeled mice ( Figure 14 A, B, and C in it). Pathological sections and H&E staining showed that most of the colonic mucosal epithelium in the compound and positive drug treatment groups proliferated and repaired, the infiltration of inflammatory cells was significantly reduced, the number of glandular cells and goblet cells increased, and the arrangement was relatively neat. Moreover, the glandular arrangement in the compound 9 treatment group was neat, similar to that of the blank group, with less infiltration of inflammatory cells, and the degree of protection of the glandular structure was better than that of the positive drug treatment group ( Figure 14 D in it).

[0178] The docking results of compounds 9 and 13 with the NLRP3 inflammasome protein showed that compound 9 had strong hydrogen - bond interactions with the residues of Phe236, Asn242, Tyr255, and Pro256 ( Figure 15 A in it), and compound 13 had hydrogen - bond interactions with the residues of Ser447 and Gln450 ( Figure 15 B in it). From this docking result, it can be inferred that the residues of Phe236, Asn242, Tyr255, Pro256, Ser447, and Gln450 in the NLRP3 inflammasome protein are the active sites of limonoids and the NLRP3 inflammasome.

[0179] The docking results of compound 9 and 13 with IL-17A protein showed that compound 9 had hydrogen bond interactions with Val65, Gln94, Glu95 and Lys114 residues ( Figure 15 C in), and compound 12 had strong hydrogen bond interactions with Thr48, Cys76, Asn78 and Ala79 residues ( Figure 15 D in). From the docking results, it can be inferred that Thr48, Val65, Cys76, Asn78, Ala79, Gln94, Glu95 and Lys114 residues in IL-17A protein are the active sites of limonin and NLRP3 inflammasome.

[0180] It is easy for those skilled in the art to understand that the above are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A tetracyclic triterpenoid compound, characterized in that, The structural formula of the tetracyclic triterpenoid compounds is as follows: Among them, R in formula VII is selected from hydroxyl or methoxy; R in formula VIII is selected from α-hydroxy or β-hydroxy; R1 and R2 in formula IX are each independently selected from hydroxyl or carbonyl; R1 and R2 in formula X are each independently selected from α-hydrogen, β-hydrogen, hydroxyl or methoxy; R in formula XI is selected from hydroxyl or hydrogen; R1 and R2 in formula XIV are each independently selected from hydroxyl or carbonyl.

2. The tetracyclic triterpenoid compound according to claim 1, wherein Formula I is compound 1, formula II is compound 2, formula III is compound 3, formula IV is compound 4, formula V is compound 5, formula VI is compound 6, formula VII is compound 7 or compound 8, formula VIII is compound 9 or compound 10, formula IX is compound 11 or compound 12, formula X is compound 13, compound 14 or compound 15, formula XI is compound 16 or compound 17, formula XII is compound 18, formula XIII is compound 19, formula XIV is compound 20 or compound 21, formula XV is compound 22, formula XIV is compound 23; 3. The separation method of the tetracyclic triterpenoid compound according to claim 2, characterized in that, It includes the following steps: (1) Dry and powder the bark of Enshi Phellodendron chinense Schneid., extract it with ethanol, and concentrate it under reduced pressure to obtain a total extract. (2) Suspend the total extract obtained in step (1) in water, extract it with chloroform and ethyl acetate respectively, and then concentrate it under reduced pressure to obtain chloroform and ethyl acetate part extraction extracts respectively. (3) Dissolve the chloroform part extraction extract obtained in step (2) with a mixed solvent of dichloromethane and methanol, mix it with silica gel and then load it onto a silica gel column, perform gradient elution with a dichloromethane and methanol system, and combine with normal-phase thin-layer chromatography TLC color development to divide it into 12 components A1 - A12; dissolve the ethyl acetate part extraction extract obtained in step (2) with a mixed solvent of dichloromethane and methanol, mix it with silica gel and then load it onto a silica gel column, perform gradient elution with a dichloromethane and methanol system, and combine with normal-phase thin-layer chromatography TLC color development to divide it into 8 components B1 - B8; When component A3 is separated by gradient elution through an ODS reverse-phase column chromatography to obtain 17 sub-components A3a - A3q; among them, A3l is separated into 5 sub-components A3l1 - A3l5 by gel column chromatography, and among them, A3l2 is further purified by semi-preparative HPLC to obtain compound 19; When component A4 is separated by gradient elution through an ODS reverse-phase column chromatography to obtain 16 sub-components A4a - A4p; Among them, A4f is separated into 3 sub-components A4f1 - A4f3 by gel column chromatography, A4f3 is separated into 4 sub-components A4f3a - A4fd by normal-phase column chromatography, and A4fd is further purified by semi-preparative HPLC to obtain compound 21 and compound 20; When component A4h is separated into 3 sub-components A4h1 - A4h3 by gel column chromatography, and A4h3 is recrystallized to obtain compound 16; When component A4i is separated into 3 sub-components A4i1 - A4i3 by gel column chromatography, and A4i3 is purified by semi-preparative HPLC to obtain compound 18; When component A5 was separated by gradient elution through ODS reversed-phase column chromatography to obtain 14 sub-components A5a - A5n; Among them, A5h was separated by normal-phase silica gel column to obtain 5 sub-components A5h1 - A5h5, and among them, A5h3 was recrystallized to obtain compound 17; When component A5j was separated using a normal-phase silica gel column to obtain 10 sub-components A5j1 - A5j10; Among them, A5j5 was separated by gel column chromatography into 3 sub-components A5j5a - A5j5c, and among them, A5j5a was recrystallized to obtain compound 9; When component A5j5c was separated by semi-preparative HPLC to obtain compound 10; When component A5j7 was separated by gel column chromatography to obtain compound 22; When component A7 was separated by normal-phase silica gel column chromatography combined with TLC color development to obtain 9 sub-components A7a - A7i; A7e was separated by gel column chromatography into 3 sub-components A7e1 - A7e3, and among them, A7e2 was further separated by normal-phase silica gel column into 5 sub-components A7e2a - A7e2e, and among them, A7e2e was recrystallized to obtain compound 3; When component A7f was separated by normal-phase silica gel column to obtain 5 sub-components A7f1 - A7f5, and among them, A7f3 was separated by reversed-phase medium-pressure column to obtain 6 sub-components A7f3a - A7f3f, and among them, A7f3f was purified by semi-preparative HPLC to obtain compound 11 and compound 12; When component A9 was separated by reversed-phase medium-pressure column chromatography combined with TLC color development to obtain 19 sub-components A9a - A9s; Among them, A9j was separated by gel column chromatography to obtain 3 sub-components A9j1 - A9j3, and among them, A9j2 was purified by normal-phase silica gel column to obtain compound 13; When component A9m was separated by gel column chromatography into 4 sub-components A9e1 - A9e4, and among them, the combination of A9e2 and A9e3 was purified by normal-phase silica gel column and gel column for the main component respectively, and then further purified by semi-preparative HPLC to obtain compound 6; When components A10 - A11 were combined and dissolved in pure methanol, the chlorophyll components were removed by MCI column, and then gradient elution was carried out through ODS reversed-phase column chromatography into 17 sub-components A10a - A10q; among them, A10i was separated by gel column chromatography into 2 sub-components A10i1 - A10i2, and among them, A10i1 was further purified by normal-phase silica gel column and semi-preparative HPLC to obtain compound 1; When component B4 was separated by ODS reversed-phase column chromatography combined with the TLC color development situation to obtain 12 sub-components B4a - B4l; among them, B4h was first purified by gel column to obtain the main component, and then further purified by HPLC to obtain compound 5; When component B4i was purified by gel column chromatography and semi-preparative HPLC to obtain compound 8; When component B4l was separated by gel column chromatography to obtain 4 sub-components B4l1 - B4l4, according to the TLC color development results, B4l2 was separated by normal-phase silica gel chromatography column to obtain 7 sub-components B4l2a - B4l2g, and among them, B4l2c was purified by semi-preparative HPLC to obtain compound 14 and compound 15; When component B5 was separated by ODS reverse-phase column chromatography, 9 sub-components B5a - B5i were obtained; Among them, B5d was separated into 2 sub-components B5d1 - B5d2 by a gel column. Among them, B5d2 was separated into 9 sub-components B5d2a - B5d2g by normal-phase silica gel column in combination with the development of thin-layer chromatography. Among them, compound 2 was further purified from B5d2f by semi-preparative HPLC; When component B6 was separated by gradient elution of ODS reverse-phase column chromatography, 7 sub-components B6a - B6g were obtained; Among them, B6d was separated into 2 sub-components B6d1 - B6d2 by a gel column. Among them, component B6d2 was separated into 6 sub-components B6d2a - B6d2f by medium-pressure reverse-phase column. Compound 23 and compound 7 were obtained by purifying B6d2d by semi-preparative HPLC; When component B7 was separated by ODS reverse-phase column chromatography, 9 sub-components B7a - B7i were obtained; among them, B7e was separated into 7 sub-components B7e1 - B7e7 by normal-phase column chromatography. The main component of B7e6 was repeatedly purified by a gel column and then monomer compound 4 was obtained by semi-preparative HPLC.

4. Use of the tetranor - triterpenoid compound according to claim 1 or 2 in the preparation of an anti-inflammatory drug.

5. The application according to claim 4, characterized in that, The anti-inflammatory drug is a drug for treating pneumonia, a drug for treating hepatitis or a drug for treating ulcerative colitis.

6. The application according to claim 4, wherein The tetranor - triterpenoid compound is used to inhibit the expression of inflammatory factors IL-1β and IL-17A.

7. The application according to claim 4, characterized in that, The tetranor - triterpenoid compound is used to inhibit the production and secretion of inflammatory factor IL-6 in Th17 cells.

8. The application according to claim 4, wherein The tetranor - triterpenoid compound is used to reduce the expression levels of Caspase-1 and NLRP3 proteins in colon tissues.

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