Preparation and application of two iridoid compounds with anti-inflammatory activity extracted from philippine flemingia rhizome

Abstract

The invention relates to two iridoid compounds Xylanchinoid A (1) and Xylanchinoid B (2) with anti-inflammatory activity, which are extracted from philippine flemingia rhizome, and a preparation method and application thereof, and belongs to the technical field of medicines. The compound 1 and the compound 2 have an inhibition effect on cell factors TNF-alpha, IL-6 and IL-1beta secreted by LPS-induced macrophages RAW264.7, can form a pharmaceutical composition with a pharmaceutically acceptable carrier, and can be used for development of anti-inflammatory drugs. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of medicines, and specifically relates to the preparation and application of two iridoid compounds with anti-inflammatory activity extracted from the rhizome of Psoralea corylifolia. Background Art

[0002] Xylanche himalaica Hook.f.et Thoms., also known as Dingzuocao, loquat taro (Shaanxi), and Pinellia (Tibet), is a plant of the genus Dingzuo of the Orobanchaceae family. It often parasitizes on the roots of plants of the genus Rhododendron and is mainly distributed in Yunnan, Qinghai, Tibet, Sichuan, Hubei and other regions. Its spherical tubers are used as medicine, which have the effects of relieving cough and expectoration, relieving swelling and strengthening the stomach, and regulating qi and relieving pain. Some ethnic minorities use its tubers to treat schistosomiasis, irregular menstruation, rheumatic joint pain, mumps, and aconite poisoning. This traditional Chinese medicine has good clinical value in traditional medicine and modern drug discovery, but its material basis research work is very weak. The main secondary metabolites isolated from this plant by domestic and foreign scholars are triterpenes, phenylethanol glycosides and lignans. No cyclopentane components have been reported from this plant, and the pharmacological activity research is relatively scarce. In order to explore the drug value of Xylanche himalaica, the present invention takes the active part of Xylanche himalaica as the research object, uses silica gel, RP 18 , MCI and other chromatographic methods were used to separate and purify the compounds. Two new cyclopentadiene ether terpenoid compounds were identified using various spectral techniques such as NMR, HR-ESI-MS, IR, etc. The pharmacological activity screening showed that the two compounds had inhibitory effects on cytokines TNF-α, IL-6 and IL-1β secreted by LPS-induced macrophages RAW264.7, and can be used for the preparation of anti-inflammatory drugs. So far, there are no reports of compounds XylanchinoidA (1) and Xylanchinoid B (2) in the prior art, nor are there any reports on their medicinal activities, let alone reports on the anti-inflammatory effects of compounds 1-2 and their pharmaceutical compositions. Summary of the invention

[0003] The purpose of the present invention is to provide a new class of two new iridoid compounds with medicinal value and their preparation methods and applications. The compounds have good anti-inflammatory activity and can be used to prepare anti-inflammatory drugs. In the in-depth study of the rhizomes of the Chinese angelica, the applicant discovered two previously unreported iridoid compounds. Therefore, the present invention provides their separation methods, structural identification, anti-inflammatory activity, medicinal prospects, etc. In order to achieve the above-mentioned purpose of the present invention, the present invention provides the following technical solutions:

[0004] The present invention provides two new iridoid compounds, the structural formulas of which are shown below:

[0005]

[0006] The two cyclopentadiene ether terpenoid compounds are both separated from the rhizomes of Xylanche himalaica, and the Xylanche himalaica used is Xylanche himalaica Hook.f.et Thoms, a plant of the Orobanchaceae family.

[0007] The preparation method of the two new iridoid compounds of the present invention comprises the following steps:

[0008] Step 1: Extraction

[0009] The dried Radix Angelicae Pubescentis material is crushed and then extracted with water by reflux. The amount of extraction solvent is 8 to 15 times the mass of the raw material. The number of extractions is 2 to 4 times, each time for 1 to 3 hours. The extract is filtered through a 200-mesh filter. The water extracts are combined and concentrated to a clear paste with a relative density of 0.8 to 1.5 (60°C). The extract is cooled and 95% ethanol is added to make the alcohol content reach 65% to 75%. The extract is allowed to stand for 24 hours and filtered. The filtrate is concentrated to obtain a fluid extract.

[0010] Step 2: Segmentation

[0011] The fluid extract obtained in step 1 was separated by silica gel column chromatography, and gradient elution was performed using chloroform: methanol solvent system of 15:1, 9:1, 8:2, 7:3, 6:4, and 1:1 as eluents in sequence. The eluate was collected and combined for TLC monitoring and divided into 11 sections (Fr1~Fr11).

[0012] Step 3: Separation and purification

[0013] The extract of Fr7 was successively purified by MCI column chromatography (water-70% methanol), silica gel column chromatography (dichloromethane: methanol: water 7:2.5:1, 7:3:1, 6:4:1, v / v / v), RP 18 Compound 1 and compound 2 were obtained by column chromatography (15%-70% methanol) gradient elution, silica gel column chromatography with dichloromethane: methanol: water 7:2.5:1 elution, silica gel column chromatography (ethyl acetate: ethanol 10:1) and Sephadex LH-20 (methanol) purification.

[0014] The present invention provides a pharmaceutical composition, which comprises at least one of the iridoid compounds with anti-inflammatory activity in the rhizome of the Chinese hollyhock and a pharmaceutically acceptable carrier; the pharmaceutical composition is divided into oral, external or injectable pharmaceutical compositions according to the administration route, and the present invention has no special limitation on the dosage form, and dosage forms well known in the art can be selected, including but not limited to tablets, capsules, granules, powders, oral liquids, sprays, plasters, powder injections or injections.

[0015] The present invention also provides the use of the iridoid compound or the pharmaceutically acceptable salt of the compound with anti-inflammatory activity in the said weight or the said pharmaceutical composition in the preparation of anti-inflammatory drugs.

[0016] The embodiments of the present invention provide the in vitro anti-inflammatory activities of the two new iridoid compounds.

[0017] Beneficial effects of the present invention:

[0018] The present invention reports two new iridoid compounds, Xylanchinoid A (1) and Xylanchinoid B (2), from the rhizome of Xylanchinoid. Compounds 1 and 2 have an inhibitory effect on the cytokines TNF-α, IL-6 and IL-1β secreted by LPS-induced macrophages RAW264.7, and can be combined with a pharmaceutical carrier to form a pharmaceutical composition for the preparation of anti-inflammatory drugs. The present invention further enriches the chemical substance basis of the active site of Xylanchinoid, provides active lead compounds for the development of new drugs, and also provides a theoretical basis for the research and development of the drugability of Xylanchinoid. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 These are the cytotoxicity test results of compound 1 and compound 2.

[0020] Figure 2 These are the test results of inflammation-related indicators of compound 1 and compound 2.

[0021] Figure 3 For compound 1 1 H- 1 H COSY and HMBC are correlated.

[0022] Figure 4 Comparison of the experimental and theoretical ECD of compound 1.

[0023] Figure 5 For compound 2 1 H- 1 H COSY and HMBC are correlated. DETAILED DESCRIPTION

[0024] In order to better understand the essence of the present invention, the following is a description of the iridoid compounds, Xylanchinoid AB (1-2) and their preparation methods, structural identification, and pharmacological effects in combination with the accompanying drawings, but the present invention is not limited to these test examples and examples.

[0025] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] Example 1

[0027] Isolation and preparation of compound 1 and compound 2

[0028] Step 1: Extraction

[0029] 34 kg of rhizome of Psoralea corylifolia was crushed into coarse powder by CSJ-60 coarse crusher and divided into 2 batches, 17 kg / batch. Each batch was decocted twice with water, 10 times (170 L) of water was added each time, and decocted for 1.5 h. The extract was filtered through a 200-mesh filter, and the water extracts were combined and concentrated to a clear paste with a relative density of 1.1 (60°C). After cooling, 95% ethanol was added to make the alcohol content reach 70%, and the mixture was allowed to stand for 24 hours. After filtering, the filtrate was concentrated to obtain 13 kg of fluid extract.

[0030] Step 2: Segmentation

[0031] Take 2 kg of the above-mentioned Qianjinzhui extract, mix it with silica gel (100-200 mesh) in a mass ratio of 1:1.2, separate it by silica gel (200-300 mesh) column chromatography, and use 15:1→1:1 chloroform-methanol as the eluent for gradient elution. Collect each fraction separately, and combine the same fractions under TLC monitoring to obtain 11 component segments, which are named Fr1~Fr11 respectively.

[0032] Step 3: Separation and purification

[0033] The extract of Fr7 (324.51 g) was eluted by MCI column chromatography (water-70% methanol) and the same fractions were combined to obtain 10 fractions, Fr7.1-Fr7.10. Fr7.3 (200 g) was eluted by silica gel column chromatography (dichloromethane: methanol: water 7:2.5:1, 7:3:1, 6:4:1, v / v / v) and the same fractions were combined to obtain 8 fractions, Fr7.3.1-Fr7.3.8. Fr7.3.5 (40 g) was eluted by RP 18Column chromatography (15%-70% methanol) was used as gradient elution and TLC detection was performed. The same fractions were combined to obtain 9 sub-fractions: Fr7.3.5.1-Fr7.3.5.9. Fr7.3.5.3 (4.6 g) was eluted with dichloromethane:methanol:water 7:2.5:1 by silica gel column chromatography, and the same fractions were combined under TLC monitoring to obtain Fr7.3.5.3.1-Fr7.3.5.3.6; Fr7.3.5.3.1 (450 mg) was purified by sephadex LH-20 (methanol), and the same fractions were combined under TLC monitoring to obtain 2 segments, Fr7.3.5.3.1.1- Fr7.3.5.3.1.2; Fr7.3.5.3.1.1 (210 mg) was purified by sephadex LH-20 (methanol) to obtain compound 2 (39.8 mg), and Fr7.3.5.3.1.2 (120 mg) was purified by silica gel column chromatography with ethyl acetate:ethanol 10:1 to obtain compound 1 (25 mg).

[0034] Example 2

[0035] Isolation and preparation of compound 1 and compound 2

[0036] Step 1: Extraction

[0037] The rhizome of Qianjinzhui is crushed into coarse powder, placed in a multifunctional extraction tank, and 8 times the amount of water is added for reflux extraction. The extract is decocted for 3 hours and extracted twice in total. The extract is filtered through a 200-mesh filter, the water extract is combined, and concentrated to a clear paste with a relative density of 1.5 (60°C). It is cooled, and 95% ethanol is added to make the alcohol content reach 65%. It is allowed to stand for 24 hours, filtered, and the filtrate is concentrated to obtain a fluid extract for use.

[0038] Step 2 segmentation and step 3 separation and purification are the same as in Example 1.

[0039] Example 3

[0040] Isolation and preparation of compound 1 and compound 2

[0041] Step 1: Extraction

[0042] The medicinal material of Qianjinzhui is crushed into coarse powder, placed in a multi-functional extraction tank, 15 times the amount of water is added each time, decocted for 1 hour, extracted twice, the extract is filtered through a 200-mesh filter, the water extract is combined, concentrated to a clear paste with a relative density of 0.8 (60°C), cooled, and 95% ethanol is added to make the alcohol content reach 75%, allowed to stand for 24 hours, filtered, and the filtrate is concentrated to obtain a fluid extract for use.

[0043] Step 2 segmentation and step 3 separation and purification are the same as in Example 1.

[0044] Example 4

[0045] Structural identification of compounds 1 and 2

[0046] (1) Structural data of compound 1.

[0047]

[0048] Xylanchinoid A (1)

[0049] Molecular formula: C 15 H 20 O9

[0050] Appearance: White solid

[0051] Optical rotation: 109.41 (c 0.170, methanol)

[0052] HRESIMS (+) m / z: Found 389.1099 [M + COOH] + , calculated value 389.1089 [M + COOH] + .

[0053] FT-IR (KBr) ν max : 3403, 2925, 1766, 1383, 1077 cm -1 .

[0054] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 1.

[0055] Compound 1, white solid, was identified by positive ion HR-ESI-MS (m / z 389.1099 [M+COOH] + , calculated value: 389.1089) The molecular formula was determined to be C 15 H 20 O9, unsaturation is 6. 1 H NMR spectrum (Table 1) showed one oxymethylene signal [δ H 4.41 (m, H-3a), 4.45 (m, H-3b)], 1 methylene [δ H 2.57 (dd , J = 9.5, 18.1 Hz, H-9a), 2.37 (dd , J = 9.5, 18.1 Hz, H-9b)], one trisubstituted double bond proton signal [δ H 5.60 (m,H-5)], 2 methine groups [δ H3.51 (m, H-10), 3.44 (m, H-11)], two oxidized methine groups [δ H 5.24 (d, J = 4.9 Hz, H-1); 5.77 (m, H-6)], and also showed a terminal proton signal belonging to the sugar fragment [δ H 4.50 (d , J = 7.9 Hz, H-1')]; 13 C NMR (Table 1) spectrum and HSQC showed 15 carbon signals (Table 1), including 2 methylene [including 1 oxidized carbon signal δ C (65.3, C-3)], 4 methines [including 2 oxidized carbon signals, δ C 105.4(C-1), 95.1(C-6)], 1 trisubstituted double bond carbon signal [δ C 120.3 (C-5), 151.3 (C-4)], and 1 lactone carbonyl [δ C 179.1 (C-8)], and the remaining 6 carbon signals can be attributed to the signals of the sugar fragment (δ C 103.6, C-1'; 75.0,C-2'; 77.8, C-3'; 71.2, C-4'; 78.5, C-5'; 62.6, C-6'). By comparing with the literature, the data are consistent with β-D-pyranose. The lactone carbonyl, double bond and sugar fragment account for 3 unsaturations, indicating that compound 1 is a tricyclic system compound. HMBC spectrum ( Figure 3 ), H-1 and C-3, C-4, C-11 (δ C 58.2) and C-1', and 1 H– 1 In the H COSY spectrum, H-1 and H-11 are related, which constructs the tetrahydrofuran ring in the compound, and C-1 is connected to a β-D-pyranose group. In the HMBC spectrum, H-5 is related to C-4, C-6, C-10 (δ C 41.3), C-11 related, H-1 and C-10 related, and 1 H– 1 In the H COSY spectrum, H-10 is correlated with H-11 and H-6, so the five-membered carbon ring in the compound is constructed and connected to the tetrahydrofuran ring through the C-4 / C-11 bond, and the double bond substitution is determined at the C-4 and C-5 positions. 13 There is one ester carbonyl signal left in the C NMR spectrum, and combined with the correlation between H-9 and C-8, C-6, C-11, and C-10 in the HMBC spectrum, 1 H– 1In the H COSY spectrum, H-10 and H-9 are correlated, indicating the presence of a five-membered lactone ring connected by a C-6 / C-10 bond. The above information can be used to infer that the planar structure of compound 1 has a five-membered carbon ring, a five-membered lactone ring, and a tetrahydrofuran ring, which belongs to the structure of an cyclopentadiene ether terpene. In the ROESY spectrum ( Figure 3 ), H-10 is related to H-6 and H-11, indicating that H-6, H-10, and H-11 are on the same side. Finally, the absolute configuration of compound 1 was determined by calculating the ECD; Figure 4 As shown in the figure, the calculated ECD spectra of 6R, 10R, 11S-1 are very consistent with the measured ECD spectra, so the absolute configuration of compound 1 is determined to be 6R, 10R, 11S. After searching, compound 1 is a novel cyclopentadiene compound, named Boschnoside A.

[0056] (2) Structural data of compound 2.

[0057]

[0058] Xylanchinoid B (2)

[0059] Molecular formula: C 16 H 20 O 10

[0060] Appearance: Amorphous powder

[0061] Optical rotation: 28.80 (c 0.150, methanol)

[0062] HRESIMS (-) m / z: Found 371.0994 [MH] – , calculated value 371.0984 [MH] – .

[0063] IR (KBr) ν max : 3418, 1740, 1103, 1057, 1041 cm -1 .

[0064] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 1.

[0065] Compound 2, a white solid, was isolated by negative ion HR-ESI-MS (m / z 371.0994 [MH] - , calculated value: 371.0984) The molecular formula was determined to be C 16 H 20 O 10 , the unsaturation is 7.1 H NMR spectrum (Table 1) showed one oxymethylene signal [δ H 4.12 (d, J = 15.1 Hz, H-10a), 4.21 (d, J = 15.1 Hz, H-10b)], one trisubstituted double bond proton signal [δ H 5.82 (s, H-7)], one sugar terminal proton signal [δ H 4.92 (d , J = 7.2 Hz, H-1')], three oxidized methine signals [δ H 5.53 (d, J = 1.6 Hz, H-1), 5.32(d, J = 2.5 Hz, H-3), 5.59(d, J = 7.8 Hz, H-6)]. 13 The C NMR spectrum (Table 1) showed 16 carbon signals, including 2 acetal signals [δ C 92.9(C-1), 96.5(C-3)], 1 carbonyl carbon signal [δ C 180.6(C-11)], 1 double bond carbon signal [δ C 126.9(C-7), 150.4(C-8)], one oxidized methylene [δ C 60.1(C-10)], one oxidized methine [δ C 88.7(C-7)], 3 methine groups [δ C 39.1(C-4), 36.1(C-5), 45.8(C-9)], and the β-D-pyranose signal [δ C 99.2(C-1),79.9(C-2), 75.9(C-3), 71.1(C-4), 79.3(C-5), 62.5(C-6)]. This compound is similar to the known compound Eucomoside A. 1 H and 13 C NMR data were compared, the only difference was that compound 2 lacked an acetyl group at the C-10 position. H 4.20) is related to C-7, C-8 and C-9, confirming the above speculation. H 3.18) and H-5 (δ H 3.59), H-5 and H-6 (δ H 5.59), H-5 and H-9 (δ H 3.16), H-1 and H-9, H-1 and H-1', H-4 and H-2' (δ H3.43), indicating that H-4 and H-6 are β-configurations; the above information, combined with H-1 (δ H 5.53, d, J = 1.6 Hz) and H-3 (δ H 5.32, d, J = 2.5 Hz), it was inferred that H-1 and H-3 were in α configuration. Therefore, the structure of compound 2 was identified and named Boschnoside A. This is the second report of an cyclopentadiene ether terpenoid with a saturated bond between C-3 and C-4 and an ether bond between C-3 and C-2' of glucose.

[0066] Table 1. Compound 1 and Compound 2 1 H (500 MHz) and 13 C NMR (125 MHz) data

[0067]

[0068] Example 5

[0069] Activity assay of compounds 1 and 2

[0070] The prepared compounds were subjected to in vitro anti-inflammatory experiments. The cells used in the experiments were mouse RAW264.7 cells purchased from the China Center for Type Culture Collection of Wuhan University. The CCK-8 method was used for cytotoxicity experiments to determine the dosage concentration, and the Elisa kit was used to detect inflammatory factors IL-6, IL-1β, and TNF-α. The specific implementation method is as follows:

[0071] Cytotoxicity experiment: Compound 1 and Compound 2 were prepared into 200, 100, 50, 25, and 12.5 μg / mL solutions using DMEM medium for later use. RAW264.7 cells were routinely cultured and grown to 80% confluence. When the cell morphology was normal under microscope, the cells were plated at 2×10 5 Cells were inoculated at a density of 100 μL / well in a 96-well plate at 100 μL / well. After overnight incubation in a carbon dioxide incubator, the supernatant in the wells was discarded, and sample solutions of different concentrations were added, 100 μL per well, 6 wells for each concentration. The blank control group was added with the same volume of DMEM and incubated in a carbon dioxide incubator. After 24 hours, 10 μL of CCK-8 reagent solution was added to each well. After 4 hours of incubation in a carbon dioxide incubator, the absorbance was detected at 450 nm using a continuous wavelength microplate reader to calculate the cell survival rate. Quantitative data are expressed as mean ± standard deviation. SPSS27.0 statistical software was used for one-way analysis of variance, and pairwise comparisons were made. The difference was considered statistically significant at p < 0.05. The results are as follows Figure 1As shown, the maximum non-toxic dose of compound 1 to RAW264.7 cells is 200 μg / mL, and the maximum non-toxic dose of compound 2 to RAW264.7 cells is 100 μg / mL.

[0072] ELISA experiment: RAW264.7 cells were routinely cultured and grown to 80% confluence. When the cells were observed to be normal in morphology under a microscope, the cells were plated at 2×10 5 Cells were inoculated at a density of 100 μg / mL in a 6-well plate, 2 mL / well, and cultured in a carbon dioxide incubator overnight. The supernatant in the wells was discarded. Sample 1 was added with 200 and 100 μg / mL sample solutions as high and low dose groups; sample 2 was added with 100 and 50 μg / mL sample solutions as high and low dose groups (both diluted with DMEM, and the concentration was confirmed by the results of the cytotoxicity test). At the same time, Lipopolysaccharide (LPS) was added for modeling, with a concentration of 200 ng / mL, 1 mL per well, and 3 wells for each concentration. The blank control group was added with the same volume of DMEM, and the model control group was added with DMEM solution containing the same concentration of LPS, and cultured in a carbon dioxide incubator. After 24 hours, the cell supernatant was aspirated to detect IL-6, IL-1β, and TNF-α kits. The experimental results are shown in the figure. Figure 2 As shown, combined with the comprehensive effects of each sample on the three inflammatory mediators, the results showed that compounds 1 and 2 could reduce the concentrations of TNF-α, IL-6 and IL-1β in the supernatant of RAW264.7 cells induced by LPS to varying degrees, suggesting that they had certain anti-inflammatory effects.

[0073] Example 6

[0074] Preparation of pharmaceutical preparations:

[0075] In the following formulation examples, conventional reagents are selected and the formulations are prepared according to conventional methods. This application example only reflects that at least one of the compounds 1-2 described in the present invention can be prepared into different formulations, and the specific reagents and operations are not specifically limited:

[0076] 1. Dissolve at least one of the compounds 1-2 with propylene glycol or polyethylene glycol, add water for injection according to conventional methods, filter, and sterilize by filling to prepare an injection solution, wherein the concentration of the injection solution is 0.5~5 mg / mL.

[0077] 2. Dissolve at least one of the compounds 1-2 with propylene glycol, polyethylene glycol, or DMSO, add a lyophilization protective agent (such as mannitol), dissolve it in sterile water for injection, stir to dissolve it, filter it aseptically, dispense it into ampoules, freeze-dry it at low temperature, and seal it aseptically to obtain a powder injection.

[0078] 3. Add an appropriate amount of excipient to at least one of compounds 1−2 to prepare a powder.

[0079] 4. Add appropriate amount of excipients to at least one of compounds 1-2, and prepare granules and tablets.

[0080] 5. Prepare at least one of compounds 1−2 into an oral solution according to a conventional oral solution preparation method.

[0081] 6. Add an appropriate amount of excipient to at least one of compounds 1−2 to prepare a capsule.

[0082] 7. Add an appropriate amount of excipient to at least one of compounds 1−2 to prepare granules.

[0083] As can be seen from the above examples, the present invention provides a compound in the rhizome of Psoralea corylifolia and its preparation method and application, a pharmaceutical composition and its application. The present invention provides two novel structural iridoid compounds, which can reduce the concentrations of TNF-α, IL-6 and IL-1β in the supernatant of RAW264.7 cells induced by LPS to varying degrees, can be combined with pharmaceutically acceptable carriers or excipients to form a pharmaceutical composition, and can be used to prepare anti-inflammatory drugs.

[0084] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Two iridoid compounds extracted from the rhizomes of the herb, characterized in that: The structural formula is as follows: 。 2. The method for preparing two iridoid compounds extracted from the rhizomes of the herb Psoralea corylifolia according to claim 1, characterized in that: The following steps are involved: Step 1 Extraction: After the dried Qianjinzhui medicinal material is crushed, it is extracted with water reflux, the amount of extraction solvent is 8 to 15 times the mass of the raw material, the number of extractions is 2 to 4 times, each time for 1 to 3 hours, the extract is filtered through a 200-mesh filter, the water extract is combined, and concentrated to a clear paste with a relative density of 0.8 to 1.5, cooled, and 95% ethanol is added to make the alcohol content reach 65% to 75%, and it is allowed to stand for 24 hours, filtered, and the filtrate is concentrated to obtain a fluid extract; Step 2: Segmentation: Take the fluid extract obtained in step 1, perform silica gel column chromatography, use chloroform: methanol solvent system 15:1, 9:1, 8:2, 7:3, 6:4, 1:1 as eluent for gradient elution, collect the eluate, monitor and merge it into 11 segments Fr1~Fr11 by TLC; Step 3: separation and purification: take the extract of Fr7 segment in step 2 and perform gradient elution on MCI column chromatography with water-70% methanol as eluent, then perform gradient elution on normal phase silica gel column chromatography with dichloromethane:methanol:water system volume ratios of 7:2.5:1, 7:3:1, and 6:4:1 as eluents, perform gradient elution on RP18 column chromatography with 15%-70% methanol as gradient elution, finally perform purification on silica gel column chromatography with ethyl acetate:ethanol volume ratio of 10:1 to obtain compound 1, and perform purification on Sephadex LH-20 column chromatography with methanol elution to obtain compound 2.

3. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the two iridoid compounds 1-2 or salts extracted from the rhizomes of the Herba Lycopodii of claim 1 and a pharmaceutically acceptable carrier.

4. The pharmaceutical composition according to claim 3, characterized in that The pharmaceutical composition is in the form of an edible pharmaceutical preparation.

5. The pharmaceutical composition according to claim 4, characterized in that The pharmaceutical preparation is selected from the form of tablets, capsules, granules, powders, oral solutions, sprays, plasters, powder injections or injection solutions.

6. Use of the two iridoid compounds 1-2 extracted from the plumb bodhi tree as claimed in claim 1 or the pharmaceutical composition as claimed in any one of claims 4-5 in the preparation of anti-inflammatory drugs.

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