Sesquiterpenoid penicipenoid E and preparation method and application thereof

Abstract

The invention discloses a sesquiterpenoid compound penicipenoid E, and the structural formula of the sesquiterpenoid compound penicipenoid E is shown as follows: # imgabs0 #. The invention also discloses an application of the sesquiterpenoid compound penicipenoid E in preparation of an anti-inflammatory drug. The preparation method disclosed by the invention is simple, and the compound penicipenoid E prepared by the method can play a role in resisting inflammatory activity and protecting tissues from inflammatory injury. The invention provides a new lead compound for researching and developing new anti-inflammatory drugs, and provides a scientific basis for developing and utilizing marine medicinal resources in China.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to a sesquiterpenoid compound, penisipenoid E, and a preparation method and application thereof. Background Art

[0002] Dysidea sp. XD140406D is a sponge belonging to the family Dysideidae, class Demospongiae, and Dictyoceratida. Microorganisms in some special habitats, such as those living in the deep sea or in symbiosis with animals and plants in special habitats, usually have special biosynthetic pathways to produce a series of secondary metabolites with good activity in order to cope with competing organisms or survive better when facing environmental pressures. They can produce many new compounds with unique structures and significant biological activities, including milestone discoveries of great significance for drug development. The marine environment has its own characteristics, such as high salt, high pressure, low oxygen, low temperature, weak light, oligotrophy, etc., which lead to the production of relatively unique metabolites in the ocean [Zhang Shi, Zhang Ying, Ning Yaodong, et al. Research progress on the anti-inflammatory effect of secondary metabolites of marine microorganisms [J]. Chinese Journal of Marine Drugs, 2023, 42(05).]. Sponges and their symbiotic microorganisms are important sources of many marine natural products. There is a lot of evidence that the actual producers of many novel and active compounds obtained from sponges may be their symbiotic microorganisms [Thomas T, Nakashima Y, Egami Y, et al. Metagenomic Analysis of the Sponge Discodermia Reveals the Production of the Cyanobacterial Natural Product Kasumigamide by 'Entotheonella' [J]. Plos One, 2016, 11 (10): e0164468. Jordan PA, Moore BS. Biosynthetic Pathway Connects Cryptic Ribosomally Synthesized Posttranslationally Modified Peptide Genes with Pyrroloquinoline Alkaloids [J]. Cell Chemical Biology, 2016, 23 (12): 1504-1514.].

[0003] Penicillium is a widely distributed and very common microorganism. A large part of Penicillium that produces good metabolites comes from the ocean. According to a survey of marine microbial natural products from 2010 to 2013 [Zhao CY, Zhu TH, Zhu W M. New marine natural products of microbial origin from 2010 to 2013 [J]. Chinese JOrg Chem, 2013, 33 (6): 1195-1234.], 22% of natural products from marine fungi were found from Penicillium, and Penicillium plays an important role in the discovery of active natural products from marine fungi. There are many types of active secondary metabolites produced by Penicillium fungi, mainly terpenes [Zou JB, Wu JL, Ding LJ, et al. Guignardones YZ, antiviral meroterpenes from Penicillium sp. NBUF154 associated with a Crella sponge from the marine mesophotic zone [J]. ChemBiodivers, 2022, 19(8). Meng LH, Li XM, Li HL, et al. Chermebilaenes A and B, new bioactive meroterpenoids from co-cultures of marine-derived isolates of Penicillium bilaiae MA-267 and Penicillium chermesinum EN-480 [J]. MarDrugs, 2020, 18(7): 339.], alkaloids [Yao FH, Liang X, Lu XH, et al. Pyrrospirones K–Q, decahydrofluorene-class alkaloids from the marine-derived fungus Penicilliumsp.SCSIO 41512[J].JNatProd,2022,85(8):2071-2081.Zhang YH,Du HF,Liu YF,etal.Novel anti-inflammatory diketopiperazine alkaloids from the marine-derivedfungus Penicillium brasilianum[J].ApplMicrobiolBiot,2024,108(1).], polyketides[WeiX, Feng C, Li XH, et al.Enantiomeric polyketides from the starfish-derivedsymbiotic fungus Penicillium sp.GGF16-1-2[J].Chem Biodivers,2019,16(6).Pang Penicillium chrysogenum DXY-1 exhibits anti-quorum sensing activity[J].Acs Omega,2021,6(11):7693-7700.Hou XM,Li YY,Shi YW,et al.Integrating molecular networking and. 1 H NMR to target the isolation of chrysogeamides from a library of marine-derived Penicillium fungi [J]. J Org Chem, 2019, 84 (3): 1228-1237.] and other compounds. Different structures give them diverse biological activities. After years of research, it has been found that the secondary metabolites of Penicillium have a variety of biological activities, including anti-cancer, anti-inflammatory, and antioxidant. Penicillium fungi have many sources and types, and their metabolites are diverse and active, making them excellent research objects.

[0004] Inflammation is an important immune defense mechanism of the body. When tissues are damaged or infected by toxins or bacteria, or damaged by heat or other reasons, they will enter a normal state of self-protection and damage repair. From the nature of inflammation, inflammation clears and eliminates various inflammatory factors and promotes the repair of damaged tissues. It is a defensive adaptive response of the body [Schultheiss C, Willscher E, Paschold L, et al. The IL-1beta, IL-6, and TNFcytokine triad is associated with post-acute sequelae of COVID-19 [J]. Cell Rep Med, 2022, 3 (6): 100663.], but in the process of anti-damage, it can also cause blood circulation disorders, release of inflammatory mediators, and tissue degeneration and necrosis. A large number of studies have shown that inflammatory response is the main response in the pathogenesis of many diseases [Meizlish ML, Franklin RA, Zhou X, et al. Tissue Homeostasis and Inflammation [J]. Annu Rev Immunol, 2021, 39: 557-581.], such as neurodegenerative diseases, Alzheimer's disease, autoimmune diseases, etc. Therefore, inhibiting proinflammatory cytokines is an effective strategy for treating inflammation-related diseases. Summary of the invention

[0005] The purpose of the invention is to provide a sesquiterpenoid compound, penisipenoid E.

[0006] Another object of the present invention is to provide a method for preparing the sesquiterpenoid compound penisipenoid E.

[0007] The third object of the present invention is to provide a use of the sesquiterpenoid compound penisipenoid E in the preparation of anti-inflammatory drugs.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0009] In a first aspect of the present invention, a sesquiterpenoid compound penicipenoid E or a pharmaceutically acceptable salt thereof is provided, the structural formula of which is shown below:

[0010]

[0011] The second aspect of the present invention provides a method for preparing the sesquiterpenoid compound penisipenoid E, comprising the following steps:

[0012] The first step is to activate the sponge-derived Penicillium sp.LHW5975 strain with PDA, cut the agar plate with the activated strain into small pieces and inoculate it on a rice culture medium, culture the sponge-derived Penicillium sp.LHW5975 with the rice culture medium, and ferment and culture it at 22-28° C. (preferably 25° C.) for 30-40 days (preferably 35 days) to obtain a rice fermentation product of Penicillium sp.LHW5975; the rice fermentation product of Penicillium sp.LHW5975 is subjected to methanol ultrasonic extraction for 3-8 times (preferably 5 times), the extract is concentrated under reduced pressure to obtain a total extract, the total extract is suspended in water, and then extracted with an equal volume of ethyl acetate for at least 3 times, the extracts are combined, and concentrated under reduced pressure to obtain an ethyl acetate extract;

[0013] The second step is to separate the ethyl acetate extract by vacuum silica gel column (VLC column) chromatography, and the obtained fractions are separated by HPLC and analyzed by TLC and then combined into 6 fractions 1-6;

[0014] In the third step, the above-mentioned fraction 2 is separated by normal medium pressure silica gel chromatography to obtain a fraction containing sesquiterpenoid compounds, which is separated by reverse phase medium pressure ODS column chromatography to obtain a fine fraction containing sesquiterpenoid compounds, and then separated by high performance liquid chromatography to obtain a sesquiterpenoid compound penisipenoid E.

[0015] In the second step, the elution system of the vacuum silica gel column (VLC column) chromatography is petroleum ether and ethyl acetate, ethyl acetate and methanol, firstly, petroleum ether and ethyl acetate are used for gradient elution, and the elution ratios of petroleum ether and ethyl acetate are: 100:0, 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, 1:1 and 0:1; then, ethyl acetate and methanol are used for gradient elution, and the elution ratios of ethyl acetate and methanol are: 100:1, 20:1, 10:1, 5:1, 2:1, 1:1 and 0:1, and the elution time is 48 hours.

[0016] In the second step, the HPLC conditions were: gradient elution of acetonitrile and water, 10% increasing to 100% in half an hour, and then eluting with 100% acetonitrile for 15 minutes.

[0017] In the third step, the conditions of the forward medium-pressure silica gel chromatography are: gradient elution using a petroleum ether-ethyl acetate system, the elution ratios of petroleum ether-ethyl acetate are: 100:0, 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, 1:1 and 0:1, and the elution is performed for 24 hours.

[0018] In the third step, the reverse phase medium pressure ODS column chromatography conditions are: using an acetonitrile-water system for gradient elution, increasing the acetonitrile ratio from 5% to 100% within 4 hours, and then eluting with 100% acetonitrile for 30 minutes.

[0019] In the third step, the conditions of the HPLC are: 25% acetonitrile-formic acid water, flow rate 2.0 mL / min, detection wavelengths 210, 220, 254, 280, 365 nm.

[0020] The third aspect of the present invention provides a use of the sesquiterpenoid compound penisipenoid E in the preparation of anti-inflammatory drugs.

[0021] Activity tests have shown that the sesquiterpenoid compound penisipenoid E can exert anti-inflammatory effects in zebrafish and can therefore be used to develop anti-inflammatory drugs.

[0022] The anti-inflammatory drug refers to at least one of the drugs directly used to prevent, diagnose, detect, protect, treat and study inflammatory reactions and directly related diseases, and is at least one of the drugs for local anti-inflammatory reactions and drugs for systemic anti-inflammatory reactions. The inflammation is at least one of degenerative inflammation, exudative inflammation and proliferative inflammation.

[0023] The anti-inflammatory local reaction drug refers to at least one of the drugs directly used to prevent, diagnose, detect, protect, treat and study local inflammatory reactions and their directly related diseases. The local inflammatory reaction refers to at least one of redness, swelling, heat, pain and functional disorder. The functional disorder can further cause at least one of local organ dysfunction on the basis of redness, swelling, heat and pain.

[0024] The anti-inflammatory systemic response drug refers to at least one of the drugs directly used to prevent, diagnose, detect, protect, treat and study inflammatory systemic responses and their directly related diseases. The inflammatory systemic response refers to at least one of fever, changes in white blood cells in the blood, changes in the mononuclear macrophage-like cell system, and changes in parenchymal organs.

[0025] Due to the adoption of the above technical solution, the present invention has the following advantages and beneficial effects:

[0026] The preparation method of the invention is simple, and the compound penicipenoid E prepared by the method can exert anti-inflammatory activity and protect tissues from inflammatory damage. The invention provides a new lead compound for researching and developing new anti-inflammatory drugs, and provides a scientific basis for developing and utilizing my country's marine medicinal resources.

[0027] The compound penisipenoid E of the invention can exert anti-inflammatory effects in zebrafish, and thus can be used for the development of preparation of anti-inflammatory drugs.

[0028] Deposit information of biological material samples:

[0029] Collection Unit: China Center for Type Culture Collection

[0030] Address: Wuhan University, Wuhan, China

[0031] Storage date: June 6, 2024

[0032] Deposit number: CCTCC NO: M20241115

[0033] Classification and nomenclature: Penicillium sp.LHW5975 BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is a key schematic diagram of the two-dimensional NMR data of penisipenoid E.

[0035] Figure 2 Schematic diagram of the comparison of the measured ECD spectrum of penisipenoid E and the calculated ECD spectra of the two configurations.

[0036] Figure 3 Schematic diagram of the antioxidant activity results of penisipenoid E in zebrafish.

[0037] Figure 4 This is a schematic diagram of the cytotoxicity results of compound penisipenoid E on the test cells RAW264.7 mouse macrophages.

[0038] Figure 5 Schematic diagram of the inhibitory activity of penisipenoid E on the release of IL-6 by macrophages.

[0039] Figure 6 Schematic diagram of the inhibitory activity of penisipenoid E on the release of TNF-α by macrophages. DETAILED DESCRIPTION

[0040] In order to explain the present invention more clearly, the present invention is further described below in conjunction with preferred embodiments. It should be understood by those skilled in the art that the following specific description is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0041] Example 1

[0042] The preparation method of compound penisipenoid E comprises the following steps:

[0043] First, the sponge-derived Penicillium sp.LHW5975 strain was activated with PDA, and the agar plate with the activated strain was cut into small pieces and inoculated on a rice medium. The sponge-derived Penicillium sp.LHW5975 was cultured with a rice medium, and fermented and cultured at 25°C for 35 days to obtain a rice fermentation product of Penicillium sp.LHW5975; the rice fermentation product of Penicillium sp.LHW5975 was subjected to methanol ultrasonic extraction for 5 times, and the extract was concentrated under reduced pressure to obtain 97g of a total extract, and the 97g total extract was suspended in 1000mL of water, and then extracted with an equal volume of ethyl acetate for 3 times, and the extracts were combined and concentrated under reduced pressure to obtain 40g of ethyl acetate extract.

[0044] The ethyl acetate extract was separated by vacuum silica gel column (VLC column) chromatography (elution system was petroleum ether and ethyl acetate, ethyl acetate and methanol, firstly gradient elution with petroleum ether and ethyl acetate, the elution ratio of petroleum ether and ethyl acetate was: 100:0, 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, 1:1 and 0:1; then gradient elution with ethyl acetate and methanol, the elution ratio of ethyl acetate and methanol was: 100:1, 20:1, 10:1, 5:1, 2:1, 1:1 and 0:1, elution time was 48 hours), and the obtained fractions were separated by HPLC (gradient elution with acetonitrile and water, 10% acetonitrile increased to 100% within half an hour, and then eluted with 100% acetonitrile for 15 minutes), and then combined into 6 fractions 1-6 after TLC analysis.

[0045] The above-mentioned fraction 2 was separated by normal medium pressure silica gel chromatography, using a petroleum ether-ethyl acetate system gradient elution (the elution ratio of petroleum ether-ethyl acetate was: 100:0, 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, 1:1 and 0:1, elution for 24 hours), to obtain a fraction containing sesquiterpenoid compounds, which was separated by reverse phase medium pressure ODS column chromatography, using an acetonitrile-water system gradient elution (acetonitrile and water gradient elution, the acetonitrile ratio increased from 5% to 100% within 4 hours, and then 100% acetonitrile elution for 30 minutes), to obtain a fine fraction containing sesquiterpenoid compounds, which was then separated by high performance liquid chromatography (25% acetonitrile-formic acid water, flow rate 2.0 mL / min, detection wavelength 210, 220, 254, 280, 365 nm), to obtain 6.5 mg of sesquiterpenoid compound penisipenoid E (C 15 H 23 O4).

[0046] The structural formula of penicipenoid E is shown below:

[0047]

[0048] The physicochemical properties and NMR data of penicipenoid E are as follows:

[0049] penicipenoid E(1):white amorphous solids; [α] D 25 +3.4(c 0.74,MeOH); UV(MeOH)λ max (logε)213(3.69)nm; ECD(c 1.88×10 -4 M,CH3OH)λ max (Δε)202(-6.1),243(-3.03)nm; IR(KBr)ν max 3374,2955,2869,1687,1641,1536,1463,1402,1369,1260,1080,1047,1009,914,756cm -1 ; 1 H(700MHz in DMSO-d6)and 13 C(150MHz)NMR,Table 1; HRESIMSm / z 267.1601[M+H] + (calcd for C 15 H 23 O4,267.1602)see Table 1.

[0050] Table 1 NMR spectrum data of compound penisipenoid E (DMSO-d6)

[0051]

[0052]

[0053] Figure 1 This is a key schematic diagram of the two-dimensional NMR data of penisipenoid E. Figure 2 The diagram is a comparison of the measured ECD (Electronic Circular Dichroism) spectrum and the two calculated ECD spectra (a and b are 1R, 6S, 7R, 10R-1a and 1S, 6R, 7S, 10S-1b) of penisipenoid E. The absolute configuration of the compound is determined to be 1R, 6S, 7R, 10R, and its structural formula is as follows:

[0054]

[0055] Example 2

[0056] The potential anti-inflammatory activity of the compound penisipenoid E prepared in Example 1 of the present invention was evaluated using the Tg (zlyz-EGFP) inflammatory cell fluorescence transgenic zebrafish model and lipopolysaccharide (LPS)-induced RAW264.7 mouse macrophages.

[0057] The compound of the present invention, penicipenoid E, was subjected to an in vivo inhibition experiment of copper sulfate-induced transgenic zebrafish inflammation, and a parallel experiment was conducted with indomethacin (a synthetic anti-inflammatory drug) as a comparison. The sample was dissolved with DMSO and stored at low temperature, and the concentration of DMSO in the final system was controlled within a range that did not affect the detection activity. Three replicate wells were set for each sample in the test. Transgenic zebrafish (Tg:zlyz-EGFP) expressing green fluorescent protein in macrophages (purchased from Shandong Academy of Sciences) were used for the activity test, and the culture conditions were carried out in accordance with the OECD (the Organization for Economic Cooperation and Development) specifications, and the zebrafish were cultured at 28±0.5°C and 14 / 10h light / no light cycle conditions. Healthy zebrafish seedlings were selected and placed in a 24-well plate, 2 mL of embryo culture medium was added to each well, 10 seedlings were grown in each well, and they were divided into 6 groups: control group (water), model group (10 μM CuSO4), positive control group (10 μM CuSO4 + 10 μM Indomethacin (purchased from Solarbio, China)), and 3 drug-added groups (10 μM CuSO4 + (10 μM, 20 μM, 40 μM) Penicipenoid E). Each zebrafish fry was photographed using a fluorescent microscope (Axio, Zoom.V16), and the number of macrophages was obtained using Image-Pro Plus software (the specific test method was based on the following references: (1) Li, QG; Chu, J.; Chen, XQ; Wang, JG; Wu, XM; Liu, KC Study on the antioxidant activity evaluation of Jujube (Ziziphus) leaf flavonoids in vitro and zebrafish (Daniorerio) with fluorescent skin. Sci. Technol. Food Ind. 2014, 35, 58-61. (2) Chen, C.-F.; Chu, C.-Y.; Chen, T.-H.; Lee, S.-J.; Shen, C.-N.; Hsiao, C.-D. Establishment of a transgenic zebrafish line for superficial skin ablation and functional validation of apoptosis modulators in vivo. PLoS One 2011,6,e20654).

[0058] The results are as follows Figure 3 As shown, Figure 3 Schematic diagram of zebrafish antioxidant activity results of penisipenoid E. A is a schematic diagram of fluorescence microscope photos of zebrafish seedlings in each group, and B is a schematic diagram of the calculation results of the number of macrophages in each group. Among them, Control represents the blank group, CuSO4 represents the model group, Indomethacin represents the positive control group, and 10μM, 20μM and 40μM represent three drug-dosing groups. As shown in A and B in the figure, the compound penisipenoid E of the present invention has inhibitory activity on the occurrence and spread of acute inflammation induced by copper sulfate in transgenic zebrafish, and its activity is weaker than that of the potent anti-inflammatory drug indomethacin, but has a good dose-dependent relationship.

[0059] In order to further verify the anti-inflammatory activity of the compound penisipenoid E of the present invention, the RAW264.7 mouse macrophage model induced by lipopolysaccharide (LPS) was used to evaluate the inhibitory activity of the compound on the release of inflammatory factors such as IL-6 and TNF-α from macrophages. It was found that penisipenoid E had a very strong inhibitory activity on the release of these two inflammatory factors.

[0060] The compound of the present invention, penicipenoid E, was tested for its inhibitory activity on the release of inflammatory factors such as IL-6 and TNF-α from RAW264.7 mouse macrophages induced by lipopolysaccharide (LPS), and a parallel experiment was performed using Quercetin (Que) as a positive control drug. The sample was dissolved in DMSO and stored at low temperature, and the concentration of DMSO in the final system was controlled within a range that did not affect the detection activity. Three replicate wells were set up for each sample in the test (the specific test method was according to the following references: Hu, TY; Zhang, H.; Chen, YY; Jiao, WH; Fan, JT; Liu, ZQ; Lin, HW; Cheng, BD. Dysiarenonefrom Marine Sponge Dysidea arenaria Attenuates ROS and Inflammation viaInhibition of 5-LOX / NF-κB / MAPKs and Upregulation of Nrf-2 / OH-1in RAW264.7Macrophages. Journal of Inflammation Research 2021, 14, 587–597.).

[0061] The cytotoxicity of compound penisicopenoid E on the test cells RAW264.7 mouse macrophages was evaluated. Figure 4 As shown, Figure 4 The figure is a schematic diagram of the cytotoxicity of compound penisipenoid E to the test cells RAW264.7 mouse macrophages. It can be seen from the figure that compound penisipenoid E does not show obvious toxicity to mouse macrophages at 6 different concentrations of 2.5, 5.0, 10, 20, 40, and 80 μM, indicating that compound penisipenoid E is safe when the concentration is less than 80 μM.

[0062] The inhibitory activity of compound penisipenoid E on the release of two inflammatory factors, IL-6 and TNF-α, from RAW264.7 mouse macrophages induced by LPS was evaluated at five concentrations of 1.25, 2.5, 5.0, 10, and 20 μM. Figure 5 As shown, Figure 5 The figure is a schematic diagram of the inhibitory activity of penisipenoid E on the release of IL-6 by macrophages. It can be seen from the figure that the compound penisipenoid E has a very strong inhibitory activity on the release of IL-6 by macrophages under five concentration conditions of 1.25, 2.5, 5.0, 10, and 20 μM, and shows an obvious dose-dependent relationship. Figure 6 The figure is a schematic diagram of the inhibitory activity of penisipenoid E on the release of TNF-α from macrophages. It can be seen from the figure that the compound penisipenoid E has very strong inhibitory activity on the release of TNF-α from macrophages at three concentrations of 1.25, 2.5, and 5.0 μM, and shows an obvious dose-dependent relationship.

[0063] In summary, the compound penisipenoid E of the present invention shows no obvious toxicity to mouse macrophages, and has very strong inhibitory activity on the release of IL-6 and TNF-α by macrophages. Therefore, the compound penisipenoid E can be used for the development of anti-inflammatory drugs.

[0064] The above is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment, it is not used to limit the present invention. Any technician familiar with this patent can make some changes or modify the technical contents suggested above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the solution of the present invention.

Claims

1. A sesquiterpenoid compound, penicipenoid E or a pharmaceutically acceptable salt thereof, characterized in that: The structural formula is as follows: penicipenoid E.

2. A method for preparing the sesquiterpenoid compound penisipenoid E according to claim 1, characterized in that: The following steps are involved: The first step is to activate the sponge-derived Penicillium sp.LHW5975 strain with PDA, cut the agar plate with the activated strain into small pieces and inoculate it on a rice culture medium, culture the sponge-derived Penicillium sp.LHW5975 with the rice culture medium, and ferment and culture it at 22-28° C. for 30-40 days to obtain a rice fermentation product of Penicillium sp.LHW5975; the rice fermentation product of Penicillium sp.LHW5975 is subjected to methanol ultrasonic extraction for 3-8 times, the extract is concentrated under reduced pressure to obtain a total extract, the total extract is suspended in water, and then extracted with an equal volume of ethyl acetate for at least 3 times, the extracts are combined, and concentrated under reduced pressure to obtain an ethyl acetate extract; The second step is to separate the ethyl acetate extract by vacuum silica gel column chromatography, and the obtained fractions are separated by HPLC and analyzed by TLC and then combined into 6 fractions 1-6; In the third step, the above-mentioned fraction 2 is separated by normal medium pressure silica gel chromatography to obtain a fraction containing sesquiterpenoid compounds, which is separated by reverse phase medium pressure ODS column chromatography to obtain a fine fraction containing sesquiterpenoid compounds, and then separated by high performance liquid chromatography to obtain a sesquiterpenoid compound penisipenoid E.

3. The method for preparing the sesquiterpenoid compound penisipenoid E according to claim 2, characterized in that: In the second step, the elution system of the vacuum silica gel column chromatography is petroleum ether and ethyl acetate, ethyl acetate and methanol. First, petroleum ether and ethyl acetate are used for gradient elution, and the elution ratios of petroleum ether and ethyl acetate are: 100:0, 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, 1:1 and 0:1; then ethyl acetate and methanol are used for gradient elution, and the elution ratios of ethyl acetate and methanol are: 100:1, 20:1, 10:1, 5:1, 2:1, 1:1 and 0:1, and the elution time is 48 hours.

4. The method for preparing the sesquiterpenoid compound penisipenoid E according to claim 2, characterized in that: In the second step, the HPLC conditions were: gradient elution of acetonitrile and water, 10% increasing to 100% in half an hour, and then eluting with 100% acetonitrile for 15 minutes.

5. The method for preparing the sesquiterpenoid compound penisipenoid E according to claim 2, characterized in that: In the third step, the conditions of the forward medium-pressure silica gel chromatography are: gradient elution using a petroleum ether-ethyl acetate system, the elution ratios of petroleum ether-ethyl acetate are: 100:0, 100:1, 50:1, 20:1, 10:1, 5:1, 2:1, 1:1 and 0:1, and the elution is performed for 24 hours.

6. The method for preparing the sesquiterpenoid compound penisipenoid E according to claim 2, characterized in that: In the third step, the reverse phase medium pressure ODS column chromatography conditions are: using an acetonitrile-water system for gradient elution, increasing the acetonitrile ratio from 5% to 100% within 4 hours, and then eluting with 100% acetonitrile for 30 minutes.

7. The method for preparing the sesquiterpenoid compound penisipenoid E according to claim 2, characterized in that: In the third step, the conditions of the HPLC are: 25% acetonitrile-formic acid water, flow rate 2.0 mL / min, detection wavelengths 210, 220, 254, 280, 365 nm.

8. Use of the sesquiterpenoid compound penisipenoid E according to claim 1 in the preparation of anti-inflammatory drugs.

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