CN116836139A - Pyrone compound based on endophytic fungus source and preparation method and application thereof - Google Patents

Abstract

The invention discloses a pyrone compound based on endophytic fungi and its preparation method and application. The pyrone compound is derived from the endophytic fungus Ascomycotasp.FAE17 strain of Scutellaria baicalensis and is prepared through the fermentation of the strain. The fermentation product is extracted with ethyl acetate, and the extract is concentrated under reduced pressure to obtain an ethyl acetate extraction extract, which is separated and purified by column chromatography to obtain pyrone compounds, which are compound 1, compound 2, compound 3 and/or compound 4 respectively. Various in vitro activity evaluation results of the pyrone compounds show that the above pyrone compounds have antibacterial and anti-inflammatory activities, can be further developed into anti-inflammatory drugs, and have the prospect of being developed into anti-inflammatory drugs, especially in the inhibition of Application of nitric oxide production in anti-inflammatory drugs.

Description

Pyrone compounds derived from endophytic fungi and preparation methods and applications thereof

Technical field

The invention belongs to the field of natural product applications and relates to a pyrone compound and its preparation method and application. Specifically, it relates to a pyrone compound derived from endophytic fungi of Scutellaria baicalensis and its preparation method and application.

Background technique

There are about 360 species of plants in the genus Scutellaria of the family Lamiacea, most of which grow in Asia and are also distributed in Europe, North America and East Asia. There are 102 species and 50 varieties recorded in the Flora of China. Most of them are medicinal plants, which have the functions of clearing away heat and drying dampness, purging fire and detoxifying. Among them, Scutellaria baicalensis (S.baicalensis) and Scutellaria barbata (S.barbata) were selected into the 2020 edition of the "Chinese Pharmacopoeia". Modern pharmacological research shows that Scutellaria genus plants have antioxidant, anti-tumor, hepatoprotective, anti-inflammatory, anti-convulsant, antibacterial, and antiviral effects.

Blue-flowered skullcap (Scutellariaformosana) is mainly distributed in southern Hainan, eastern and southern Guangdong, Jiangxi, southern Fujian and southern Yunnan. Blue-flowered skullcap mostly grows in the shade of forests at an altitude of 450-550 meters. Its growth environment is relatively special, making it extremely rare and difficult to find, resulting in a handful of blue-flowered skullcap. Due to the limited production of Scutellaria baicalensis, it is obviously difficult to industrialize the collection of Scutellaria baicalensis plants for drug development, and it is also not conducive to the protection of rare natural medicinal plants. Therefore, there is an urgent need to develop alternative resources for medicinal Scutellaria baicalensis.

Endophytic fungi of Scutellaria baicalensis are endophytic microorganisms that live symbiotically in the host Scutellaria baicalensis. They participate in the biosynthesis of secondary metabolites of the host and metabolize rich metabolites with special structures during long-term adaptation to the natural environment. . More importantly, the endophytic fungi of Scutellaria baicalensis can be produced on a large scale through fermentation engineering technology, without destroying the scarce plant resources of Scutellaria baicalensis, making it easy to achieve industrialization. Therefore, it is of theoretical research value and important practical significance to conduct research on the secondary metabolites of endophytic microorganisms in Scutellaria baicalensis and their pharmacological activities.

The purpose of the present invention is to provide a pyrone compound derived from endophytic fungi of Scutellaria baicalensis, which has antibacterial and anti-inflammatory activities, can be further developed into an anti-inflammatory drug, and has the prospect of being developed into an anti-inflammatory drug, especially in Use in anti-inflammatory drugs that inhibit nitric oxide production.

The above objects of the present invention are achieved through the following technical solutions:

A kind of pyrone compound, described pyrone compound is compound 1, compound 2, compound 3 and/or compound 4, and its chemical structure is as follows respectively:

Another object of the present invention is to provide a method for preparing the pyrone compounds derived from endophytic fungi of Scutellaria baicalensis, which includes the following steps:

S1: Preparation of fermentation product of Ascomycota sp.FAE17 strain;

S2: Extract the fermentation product obtained in step S1 with an equal volume of ethyl acetate, and concentrate the extract under reduced pressure to obtain an ethyl acetate extraction extract;

S3: The ethyl acetate extraction extract obtained in step S2 is separated and purified by column chromatography to obtain compound 1, compound 2, compound 3 and compound 4.

Further, the step S1 includes the following steps: transfer the Ascomycota sp. FAE17 strain to PDA culture medium for culture, then transfer it to an Erlenmeyer flask containing potato glucose liquid culture medium, and shake and culture it to obtain the seed bacteria. liquid, transfer the seed bacterial liquid to a high-temperature sterilized rice culture medium Erlenmeyer flask and let it stand for fermentation to obtain a fermentation product.

Further, the step S3 includes the following steps:

(1) The ethyl acetate extraction extract is roughly separated by silica gel column chromatography, and the volume ratios are 9:1, 8:2, 7:3, 6:4, 5:5, 3:7, 2:8, respectively. Petroleum ether-ethyl acetate gradient elution was performed at 1:9, 0:1 and ethyl acetate-methanol was performed at a volume ratio of 9:1, 8:2, 7:3, 6:4, 5:5, 1:1. Gradient elution, collect the ethyl acetate-methanol eluates with volume ratios of 9:1, 8:2, and 7:3;

(2) Combine the ethyl acetate-methanol eluates with a volume ratio of 9:1, 8:2, and 7:3 and perform reverse-phase silica gel column chromatography, using a volume ratio of 1:9, 2:8, and 3: 7. Methanol-water gradient elution of 4:6, 6:4, 8:2, and 10:0, and collect the methanol-water eluate with a volume of 1:9;

(3) Subject the obtained methanol-water eluate to normal phase silica gel column chromatography, and use ethyl acetate with a volume ratio of 9:1, 8:2, 7:3, 6:4, 5:5, 3:7. -Methanol gradient elution, collect the ethyl acetate-methanol eluates with volume ratios of 9:1 and 8:2 respectively for concentration;

(4) Take the concentrated ethyl acetate-methanol eluate with a volume ratio of 9:1 and separate it with semi-preparative high-performance liquid chromatography. The mobile phase is acetonitrile-water with a volume ratio of 8:92 to obtain compound 1 and compound 1. 3 and compound 4;

(5) Take the concentrated ethyl acetate-methanol eluate with a volume ratio of 8:2 and perform reverse-phase silica gel column chromatography, using a volume ratio of 1:9, 2:8, 3:7, 4:6, Methanol-water gradient elution of 6:4, 8:2, and 10:0 was used to collect the methanol-water eluate with a volume of 3:7;

(6) The concentrated ethyl acetate-methanol eluate with a volume ratio of 3:7 was separated by semi-preparative high-performance liquid chromatography. The mobile phase was acetonitrile-water with a volume ratio of 13:87 to obtain compound 2.

Another object of the present invention is to provide the application of the pyrone compounds (Compound 1, Compound 2, Compound 3 and/or Compound 4) in anti-inflammatory aspects, especially in the preparation of anti-inflammatory drugs, and more specifically is used in anti-inflammatory drugs that inhibit nitric oxide production.

Compared with the prior art, the beneficial effects of the present invention are:

The present invention prepares ethyl acetate extraction extract from the fermentation material of endophytic fungi of Scutellaria baicalensis, and then isolates and identifies four pyrone compounds with novel chemical structures, namely compound 1, compound 2, compound 3 and compound 4. . Various in vitro activity evaluation results show that compound 1, compound 2, compound 3 and compound 4 have antibacterial and anti-inflammatory activities and can be further developed into anti-inflammatory drugs. They have the prospect of being developed into anti-inflammatory drugs, especially in inhibiting nitric oxide. Generative applications in anti-inflammatory drugs.

Description of the drawings

Figure 1 is a 1H-1H COSY and HMBC correlation diagram of compound 1, compound 2, compound 3 and compound 4 of the present invention.

Figure 2 is a NOESY correlation diagram of compound 1, compound 2, compound 3 and compound 4 of the present invention.

Figure 3 is an ECD chart of compound 1 of the present invention.

Figure 4 is an ECD chart of compound 2 of the present invention.

Figure 5 is the experimental results of anti-inflammatory activity of Compound 1, Compound 2, Compound 3 and Compound 4 of the present invention.

Detailed ways

In order to better understand the technical content of the present invention, specific examples are provided below to further illustrate the present invention. The following examples are used to illustrate the invention but are not intended to limit the scope of the invention. Experimental methods that do not indicate specific conditions in the following examples usually follow conventional experimental conditions.

Example 1. Preparation method of pyrone compounds

The endophytic fungus Ascomycota sp.FAE17 of Scutellaria baicalensis of the present invention is collected from the flower parts of the rare medicinal plant Scutellaria baicalensis in Bawangling National Nature Reserve, Changjiang Li Autonomous County, Hainan Province. The endophytic fungal strain of Scutellaria baicalensis was identified by a sequencing company (Qingdao Pengxiang Biotechnology Co., Ltd.). The obtained base sequences were compared for similarity in the GenBank database, and then the BLAST program was used to search for homologous sequences for comparison. The sequence similarity was 99%, and the FAE-17 strain was determined to be Ascomycotasp.

Take the Ascomycota sp. FAE17 strain out of the 4°C refrigerator and activate it at room temperature. Transfer it to PDA culture medium and culture it in a 28°C constant temperature incubator. Observe its growth after 2-3 days, and then transfer the strain. Put it into an Erlenmeyer flask filled with potato glucose liquid nutrient medium, place it in a room temperature shaker and shake it for 2-3 days, and observe the growth of the seed bacterial liquid; transfer the seed bacterial liquid to high-temperature sterilized rice in a clean bench In the Erlenmeyer flask of the culture medium, inoculate 150 bottles and let it stand at room temperature for 5 weeks to obtain the fermentation product; the rice solid culture medium is: 90g/bottle of rice, 120mL/bottle of water, a Erlenmeyer flask with a capacity of 1000mL, and a total of 150 bottles are inoculated. .

The fermentation product was extracted with an equal volume of ethyl acetate 3 to 5 times, the extracts were combined and concentrated under reduced pressure to obtain 160.0g of ethyl acetate extracted extract.

The ethyl acetate extraction extract was roughly separated by silica gel column chromatography, and the volume ratios were 9:1, 8:2, 7:3, 6:4, 5:5, 3:7, 2:8, 1:9, respectively. 0:1 for petroleum ether-ethyl acetate gradient elution and volume ratios of 9:1, 8:2, 7:3, 6:4, 5:5, 1:1 for ethyl acetate-methanol gradient elution, collect Ethyl acetate-methanol eluates with volume ratios of 9:1, 8:2, and 7:3;

Combine the ethyl acetate-methanol eluates with volume ratios of 9:1, 8:2, and 7:3 and perform reverse-phase silica gel column chromatography, using a volume ratio of 1:9, 2:8, 3:7, and 4. :6, 6:4, 8:2, 10:0 methanol-water gradient elution, and collect the methanol-water eluate with a volume of 1:9.

Take the methanol-water eluate with a volume of 1:9 for normal phase silica gel column chromatography, and use acetic acid with a volume ratio of 9:1, 8:2, 7:3, 6:4, 5:5, 3:7. Ethyl acetate-methanol gradient elution, the ethyl acetate-methanol eluates with volume ratios of 9:1 and 8:2 were collected and concentrated;

The concentrated ethyl acetate-methanol eluate with a volume ratio of 9:1 was separated by semi-preparative high-performance liquid chromatography. The mobile phase was acetonitrile-water with a volume ratio of 8:92 to obtain compound 1 (20.1 mg), Compound 3 (1.2mg) and Compound 4 (1.7mg);

Take the concentrated ethyl acetate-methanol eluate with a volume ratio of 8:2 and perform reverse-phase silica gel column chromatography, using a volume ratio of 1:9, 2:8, 3:7, 4:6, and 6:4. , 8:2, 10:0 methanol-water gradient elution, and collect the methanol-water eluate with a volume of 3:7.

The concentrated ethyl acetate-methanol eluate with a volume ratio of 3:7 was separated by semi-preparative high-performance liquid chromatography. The mobile phase was acetonitrile-water with a volume ratio of 13:87 to obtain compound 2 (4.6 mg).

Structural confirmation: Through comprehensive analysis using various modern spectroscopic technologies such as optical rotation spectrum, ultraviolet (UV) spectrum, infrared (IR) spectrum, nuclear magnetic resonance (NMR) spectrum, mass spectrometry (MS) and quantum chemical ECD calculation methods, the obtained ¹ HNMR and ¹³ C NMR (400/100MHz, DMSO-d ₆ ) data are shown in Table 1 and Table 2, and the chemical structures of Compound 1, Compound 2, Compound 3 and Compound 4 were determined:

Compound 1: light yellow oil; UV(MeOH)λ _max [logε/(L·mol ^-1 ·cm ^-1 )]:249(2.30)nm; IR(KBr)ν _max :1710cm ^-1 ,1622cm ^-1 ; HR-ESI-MS m/ z251.0877[M+Na] ⁺ (calcd for C ₁₁ H ₁₆ O ₅ Na,251.0890). The two-dimensional correlation signals of carbon and hydrogen are shown in Figure 1 (1H-1HCOSY, HMBC diagram). The relative configuration is shown in Figure 2. The relative configuration of compound 1 was determined through the correlation between H-5 and H-10 in compound 1, and its absolute configuration was determined through CD testing and ECD calculations. Comparing the CD test spectrum and ECD calculation spectrum of compound 1 (Figure 3), it was found that the Cotton effect is very consistent, that is, the absolute configuration of compound 1 is 5R, 8R and named ascomycopyroneA.

Compound 2: yellow oil; UV(CH ₃ OH)λ _max [logε/(L·mol ^-1 ·cm ^-1 )]:249(2.66),300(0.43)nm; IR(KBr)ν _max :1710cm ^-1 ,1621cm ^-1 ; HR-ESI-MSm/z 391.1349[M+Na] ⁺ (calcd for C ₁₈ H ₂₄ O ₈ Na,391.1349). The two-dimensional correlation signals of carbon and hydrogen are shown in Figure 1 (1H-1HCOSY, HMBC diagram). The relative configuration is shown in Figure 2, which is related to H-5 (δH 2.49) and H-10 (δH 0.93) in compound 2, and H-5′ (δH 2.86/4.38) and H-10′ (δH 15.7) It is speculated that there are four relative configurations of compound 2. The structure DP4+ calculation was performed, and the results showed that the configurations of 5R, 8R, 5R, and 8R-20 were completely consistent. Compound 2 was further determined through the ECD calculation spectrum (Figure 4). The absolute configuration was named ascomycopyrone B.

Compound 3: colorless oil; UV(MeOH)λ _max [logε/(L·mol ^-1 ·cm ^-1 )]:241(2.45)nm; IR(KBr)ν _max :3446cm ^-1 ,1622cm ^-1 ; HR-ESI-MS m/ z337.1608[M+Na] ⁺ (calcd for C ₁₆ H ₂₆ O ₆ Na,337.1622). The two-dimensional correlation signals of carbon and hydrogen are shown in Figure 1 (1H-1HCOSY, HMBC diagram). The relative configuration is shown in Figure 2. In compound 3, H-5 (δH 2.44) is related to H-10 (δH 0.89), H-6′ (δH 0.97) is related to H-2′ (δH 4.58), H- The relative configuration of compound 3 was determined and named ascomycopyrone C.

Compound 4: colorless oil; UV(MeOH)λ _max [logε/(L·mol ^-1 ·cm ^-1 )]:248(2.29)nm; IR(KBr)ν _max :3453cm ^-1 ,1620cm ^-1 ; HR-ESI-MS m/ z337.1604[M+Na] ⁺ (calcd for C ₁₆ H ₂₆ O ₆ Na,337.1622). The two-dimensional correlation signals of carbon and hydrogen are shown in Figure 1 (1H-1HCOSY, HMBC diagram). The relative configuration is shown in Figure 2. In compound 4, H-5 (δH 2.44) is related to H-10 (δH 0.91), H-3′ (δH 1.57) and H-6′ (δH 1.34), H- The relative configuration of compound 4 was determined based on the correlation between 8′ (δH 0.99) and H-6′ (δH 1.34) and was named ascomycopyrone D.

Table 1 ¹ HNMR and ¹³ CNMR (400/100MHz, DMSO-d ₆ ) data of compound 1 and compound 2

Table 2 ¹ H NMR and ¹³ C NMR (400/100MHz, DMSO-d ₆ ) data of compound 3 and compound 4

Example 2. Preparation method of pyrone compounds

Take the Ascomycota sp. FAE17 strain out of the 4°C refrigerator and activate it at room temperature. Transfer it to potato glucose solid medium and culture it in a 28°C constant temperature incubator. Observe its growth after 2-3 days, and then inoculate the strain. Transfer it to an Erlenmeyer flask containing potato glucose liquid culture medium, place it in a room temperature shaker and shake it for 2-3 days, and observe the growth of the seed bacterial liquid; transfer the seed bacterial liquid to a high-temperature sterilizer in a clean bench Inoculate 300 bottles of rice culture medium in Erlenmeyer flasks, and cultivate at room temperature for 5 weeks to obtain the fermentation product; among them, the rice solid culture medium is: 90g/bottle of rice, 120mL/bottle of water, and an Erlenmeyer flask with a capacity of 1000mL. A total of 300 bottles were inoculated.

The obtained fermentation product was extracted 3 to 5 times with an equal volume of ethyl acetate. The extracts were combined and concentrated under reduced pressure to obtain 310.0 g of ethyl acetate extracted extract.

The ethyl acetate extraction extract was roughly separated by silica gel column chromatography, and the volume ratios were 9:1, 8:2, 7:3, 6:4, 5:5, 3:7, 2:8, 1:9, respectively. 0:1 for petroleum ether-ethyl acetate gradient elution and volume ratios of 9:1, 8:2, 7:3, 6:4, 5:5, 1:1 for ethyl acetate-methanol gradient elution, collect Ethyl acetate-methanol eluates with volume ratios of 9:1, 8:2, and 7:3;

Combine the ethyl acetate-methanol eluates with volume ratios of 9:1, 8:2, and 7:3 and perform reverse-phase silica gel column chromatography, using a volume ratio of 1:9, 2:8, 3:7, and 4. :6, 6:4, 8:2, 10:0 methanol-water gradient elution, and collect the methanol-water eluate with a volume of 1:9.

Take the methanol-water eluate with a volume of 1:9 for normal phase silica gel column chromatography, and use acetic acid with a volume ratio of 9:1, 8:2, 7:3, 6:4, 5:5, 3:7. Ethyl acetate-methanol gradient elution, the ethyl acetate-methanol eluates with volume ratios of 9:1 and 8:2 were collected and concentrated;

The concentrated ethyl acetate-methanol eluate with a volume ratio of 9:1 was separated by semi-preparative high-performance liquid chromatography. The mobile phase was acetonitrile-water with a volume ratio of 8:92 to obtain compound 1 (40.6 mg), 3 (2.3mg), 4 (3.5mg) (structure confirmation data are the same as Example 1);

Take the concentrated ethyl acetate-methanol eluate with a volume ratio of 8:2 and perform reverse-phase silica gel column chromatography, using a volume ratio of 1:9, 2:8, 3:7, 4:6, and 6:4. , 8:2, 10:0 methanol-water gradient elution, and collect the methanol-water eluate with a volume of 3:7.

The concentrated ethyl acetate-methanol eluate with a volume ratio of 3:7 was separated by semi-preparative high performance liquid chromatography. The mobile phase was acetonitrile-water with a volume ratio of 13:87 to obtain compound 2 (9.5 mg) ( The structure verification data is the same as that in Example 1).

Study on the anti-inflammatory activity of the pyrone compounds obtained in Example 1 of the present invention:

experimental method:

(1) Use the MTT colorimetric method to screen the cytotoxic activity of mouse macrophage RAW 264.7. Take 10 μL of RAW cells in the logarithmic growth phase and put them into a counting plate for counting, so that the number of cells in each well is about 5000-10000. 96 Add 100 μL PBS solution to the outer circle of the well plate, inoculate 100 μL cell solution into each remaining well, and incubate for 24 hours to allow cells to adhere to the wall and grow. Add the prepared sample solutions sequentially from high concentration to low concentration. The final concentrations in each well are 100 μmol/L, 10 μmol/L, and 1 μmol/L respectively. Add PBS solution to the blank group, and then place it in a constant-temperature carbon dioxide incubator for 48 hours. , take out the cultured cells, add 10 μL of pre-thawed MTT at room temperature to each well, put it in the incubator for 4 hours, take out the 96-well plate, pour off the supernatant, and absorb the excess liquid on paper towels, and add 150 μL DMSO to each well. , shake in the dark to dissolve the purple crystals of formazan, remove the plate cover, and measure the absorbance values at 490, 570, and 630 nm with a microplate reader. Density value (OD), process the data at a wavelength of 570 nm, and calculate the cell inhibition rate according to the formula 1-(OD sample/OD Control) × 100%.

(2) Take the cells in the logarithmic growth phase, make a cell suspension and count them, seed them in a 96-well plate at ^2x105 cells/well, place them in a 5% CO2, 37°C incubator and culture them for 24 hours. The next day, monomeric compounds that are non-toxic to RAW cells (final concentration 6.25-50 μM) and dexamethasone and an equal volume of DMSO were added to each well for pretreatment for 1 hour. Afterwards, in addition to the blank control group, LPS (2 μg/mL) was added for stimulation. , continue culturing for 24h. The cell supernatant was collected at the end of culture.

Use the Griess method to detect NO release: Before measuring the NO content, return GriessReagent I and II to room temperature, and dilute the standard with serum-free DMEM culture medium. The concentrations are: 0, 1, 2, 5, 10, and 20. , 40, 60, and 100 μM. Take 50 μL each of the standard substance and the supernatant of the sample to be tested, and add it to a new 96-well plate. Set up three duplicate holes for each sample in the standard group, blank group, LP group, sample group, and positive drug group. Add 50 μL of griessReagent I to each well in the dark, tap the 96-well plate to mix thoroughly. After reacting for 5 minutes, add GriessReagent II and shake gently to mix the liquid in the well evenly. Use a microplate reader to measure the absorbance value at 540 nm. Substitute into the nitrite labeling curve to calculate the NO content and calculate the inhibitory rate of the compound on NO production by LPS-activated RAW264.7 cells. The results are shown in Figure 5.

NO production inhibition rate %=(LPS group-sample group)/(LPS group blank group)×100%.

The anti-inflammatory activity test results showed that at 50 μM, compound 1, compound 2, compound 3 and compound 4 all had varying degrees of inhibitory effects on NO produced by LPS-induced RAW 264.7 cells, and had certain anti-inflammatory activity.

The above are only preferred embodiments of the present invention. It should be pointed out that those of ordinary skill in the art can also make several improvements and modifications without departing from the technical principles of the present invention. These improvements and modifications It should also be regarded as the protection scope of the present invention.

Claims (7)

1. A pyrone compound, characterized in that the pyrone compound is compound 1, compound 2, compound 3 and/or compound 4, and their chemical structures are as follows: 3 R ₁ =β-H, R ₂ =β-CH ₃ , R ₃ =β-CH(CH ₃ )-β-OH 4 R ₁ =α-CH ₃ , R ₂ =β-CH ₂ OH, R ₃ =α-CH _{3 .} 2. The preparation method of pyrone compounds as claimed in claim 1, characterized in that, comprising the following steps: S1: Preparation of fermentation product of Ascomycota sp.FAE17 strain; S2: Extract the fermentation product obtained in step S1 with an equal volume of ethyl acetate, and concentrate the extract under reduced pressure to obtain an ethyl acetate extraction extract; S3: The ethyl acetate extraction extract obtained in step S2 is separated and purified by column chromatography to obtain compound 1, compound 2, compound 3 and compound 4. 3. The preparation method of pyrone compounds as claimed in claim 2, characterized in that, the step S1 includes the following steps: transfer the Ascomycota sp. FAE17 strain to culture on the PDA culture medium, and then transfer Put it into an Erlenmeyer flask containing potato glucose liquid culture medium, shake it for culture, and obtain a seed bacterial liquid. Transfer the seed bacterial liquid to a high-temperature sterilized rice culture medium Erlenmeyer flask and let it stand for fermentation to obtain a fermentation product. 4. The preparation method of pyrone compounds as claimed in claim 2, characterized in that said step S3 includes the following steps: (1) The ethyl acetate extraction extract is roughly separated by silica gel column chromatography, and the volume ratios are 9:1, 8:2, 7:3, 6:4, 5:5, 3:7, 2:8, respectively. Petroleum ether-ethyl acetate gradient elution was performed at 1:9, 0:1 and ethyl acetate-methanol was performed at a volume ratio of 9:1, 8:2, 7:3, 6:4, 5:5, 1:1. Gradient elution, collect the ethyl acetate-methanol eluates with volume ratios of 9:1, 8:2, and 7:3; (2) Combine the ethyl acetate-methanol eluates with a volume ratio of 9:1, 8:2, and 7:3 and perform reverse-phase silica gel column chromatography, using a volume ratio of 1:9, 2:8, and 3: 7. Methanol-water gradient elution of 4:6, 6:4, 8:2, and 10:0, and collect the methanol-water eluate with a volume of 1:9; (3) Subject the obtained methanol-water eluate to normal phase silica gel column chromatography, and use ethyl acetate with a volume ratio of 9:1, 8:2, 7:3, 6:4, 5:5, 3:7. -Methanol gradient elution, collect the ethyl acetate-methanol eluates with volume ratios of 9:1 and 8:2 respectively for concentration; (4) Take the concentrated ethyl acetate-methanol eluate with a volume ratio of 9:1 and separate it with semi-preparative high-performance liquid chromatography. The mobile phase is acetonitrile-water with a volume ratio of 8:92 to obtain compound 1 and compound 1. 3 and compound 4; (5) Take the concentrated ethyl acetate-methanol eluate with a volume ratio of 8:2 and perform reverse-phase silica gel column chromatography, using a volume ratio of 1:9, 2:8, 3:7, 4:6, Methanol-water gradient elution of 6:4, 8:2, and 10:0 was used to collect the methanol-water eluate with a volume of 3:7; (6) The concentrated ethyl acetate-methanol eluate with a volume ratio of 3:7 was separated by semi-preparative high-performance liquid chromatography. The mobile phase was acetonitrile-water with a volume ratio of 13:87 to obtain compound 2. 5. Use of the pyrone compounds as claimed in claim 1 in anti-inflammatory aspects. 6. Application of the pyrone compound as claimed in claim 5 in the preparation of anti-inflammatory drugs. 7. Application of the pyrone compound as claimed in claim 6 in an anti-inflammatory drug for inhibiting nitric oxide production.