Alpha-pyrone mixed-source meroterpenoid dimer Hypericumonates A-C as well as preparation method and application of alpha-pyrone mixed-source meroterpenoid dimer Hypericumonates A-C

Abstract

The invention discloses an alpha-pyrone mixed source meroterpenoid dimer Hypericumonates A-C as well as a preparation method and application of the alpha-pyrone mixed source meroterpenoid dimer Hypericumonates A-C, and belongs to the technical field of medical chemistry. The alpha-pyrone mixed source meroterpenoid dimer Hypericumonates A-C has a structure as shown in formulas A-C, and the alpha-pyrone mixed source meroterpenoid dimer Hypericumonates A-C is obtained by being separated from Hypericum monogynum for the first time. The alpha-pyrone mixed source meroterpenoid dimer Hypericumonates A-C has a structure as shown in formulas A-C; the compound Hypericumonates A-C has obvious inhibitory activity on NO concentration in BV-2 cells induced by lipopolysaccharide (LPS), shows an obvious anti-inflammatory effect, has the activity higher than that of a positive control drug minocycline, can inhibit expression of iNOS and COX-2 in a dose-dependent manner, and can be applied to preparation of anti-neuroinflammation drugs.

Description

Technical Field

[0001] The invention belongs to the field of medicinal chemistry, and specifically relates to an alpha-pyrone mixed heteroterpenoid dimer Hypericumonates AC and a preparation method and application thereof. Background Art

[0002] Neuroinflammation is a condition found in the central nervous system, usually caused by infection, toxic metabolites, trauma or autoimmune stimulation. It has a protective effect, promotes neural repair and maintains internal environment stability; however, long-term neuroinflammatory activation can lead to excessive production of inflammatory mediators, resulting in neurological dysfunction and degeneration, which is associated with a variety of neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, multiple sclerosis and Huntington's disease. Therefore, the study found that drugs that inhibit neuroinflammation are of great value and significance. The neuroinflammation treatment drugs on the market, such as memantine and tramiprosate, are not ideal in treatment effect and have large side effects. As one of the important sources of innovative drug discovery, natural medicine has become a hot field for the treatment of neuroinflammation. Therefore, the discovery of new anti-neuroinflammatory drugs from natural products has attracted the attention of many medicinal chemists. α-pyrone mixed heteroterpenoid dimer is a mother core formed by two α-pyrone dimethylpyran rings through a [2+2] cyclization reaction of the terminal double bond and the intracyclic double bond or the intracyclic double bond and the intracyclic double bond. It is a very rare natural product with a unique chemical structure and good anti-neuroinflammatory activity. In the present invention, α-pyrone mixed heteroterpenoid dimer Hypericumonates AC is a 6 / 6 / 4-6 / 6 ring system structure formed by the terminal double bond of one molecule of α-pyrone dimethylpyran ring and the intracyclic double bond of another molecule of α-pyrone ring through a [2+2] cycloaddition reaction, and finally connected in the form of CC bonds. Its basic mother core is a new type of skeleton that has never been seen before, and it exhibits significant anti-neuroinflammatory activity. Hypericum monogynum, a plant of the genus Hypericum in the family Guttiferae, is rich in α-pyrone mixed heteroterpenoid compounds, and the dimers have not been reported in the relevant literature. This plant is widely distributed in the southwestern provinces of China. Its flowers, roots, and leaves can be used as medicine, with anti-rheumatic, anti-hepatitis, anti-bacterial and anti-inflammatory, treatment of bronchitis, hemostasis and myogenic effects. At present, the relevant research on the chemical components of Hypericum is mainly based on compounds such as phloroglucinol, spironolactone, flavonoids, and triterpenoids. Therefore, it is of great research value and significance to explore α-pyrone mixed heteroterpenoid dimer compounds with novel structures and anti-neuroinflammatory activity from the ethnic medicine Hypericum. Summary of the invention

[0003] The purpose of the present invention is to dig out a novel α-pyrone mixed heteroterpenoid dimer from the ethnic medicine Hypericum perforatum, and have anti-neuroinflammatory activity, and use it in the preparation of anti-neuroinflammatory drugs. Provide an α-pyrone mixed heteroterpenoid dimer and its preparation method and application, in order to achieve the above purpose, the following technical scheme is adopted:

[0004] An α-pyrone mixed heteroterpenoid dimer Hypericumonates A-C, whose structural formula is A to C:

[0005]

[0006] Right now:

[0007]

[0008] The method for preparing the α-pyrone mixed terpenoid dimer Hypericumonates A-C comprises the following steps:

[0009] Step A: taking 50 kg of dried Hypericum perforatum branches and leaves, using 150-250 kg of methanol as solvent, extracting 3-5 times at a temperature of 20-30° C., each extraction time being 4 days, and concentrating under reduced pressure to recover methanol to obtain a crude extract I;

[0010] Step B: using dichloromethane, ethyl acetate and methanol as extraction solvents, solid phase extraction is performed on the crude extract I obtained in step A to obtain three fractions: component A, component B, and component C; component A is subjected to silica gel column chromatography, and gradient elution is performed with petroleum ether and dichloromethane in a volume ratio of 100:0-0:100; different fractions are then detected using thin layer chromatography technology, and seven fractions are obtained by merging them according to their thin layer chromatography performance: Fr.1, Fr.2, Fr.3, Fr.4, Fr.5, Fr.6, and Fr.7;

[0011] Step C: fraction Fr.5 was separated by MCI column chromatography, and the eluent was gradient eluted with a volume ratio of methanol to water of 40:60 to 100:0, and the fractions were combined according to the thin layer chromatography performance to obtain 7 sub-fractions: Fr.5.A, Fr.5.B, Fr.5.C, Fr.5.D, Fr.5.E, Fr.5.F and Fr.5.G;

[0012] Step D: Fr.5.D was separated by gel LH-20 column chromatography, eluted with methanol as solvent, and four sub-fractions were obtained: Fr.5.D.1, Fr.5.D.2, Fr.5.D.3 and Fr.5.D.4. Fraction Fr.5.D.2 was further purified by gel column chromatography and recrystallization, and the elution solvent used was methanol, to obtain α-pyrone mixed heteroterpenoid dimer Hypericumonate A;

[0013] Step E: Fraction Fr.5.C was purified by gel column chromatography and silica gel column chromatography with a gradient elution of petroleum ether and ethyl acetate in a volume ratio of 20:1 to 1:1 to obtain four subfractions: Fr.5.C.1, Fr.5.C.2, Fr.5.C.3 and Fr.5.C.4; component Fr.5.C.3 was further purified by semi-preparative high performance liquid chromatography with an acetonitrile to water volume ratio of 55:45 and a flow rate of 3.0 mL / min to obtain α-pyrone mixed heteroterpene dimers Hypericumonate B and Hypericumonate C.

[0014] The method for preparing the α-pyrone mixed terpene dimer Hypericumonates A-C is characterized in that in step A, the methanol is 100% industrial methanol; and the extraction method is cold soaking extraction.

[0015] The preparation method of the α-pyrone mixed terpene dimer Hypericumonates A-C is characterized in that, in step B, fraction A is separated by silica gel column chromatography, and the concentrations of the gradient elution of petroleum ether / dichloromethane are volume ratios of 100:0, 80:20, 70:30, 60:40, 50:50, 20:80, and 0:100, respectively.

[0016] The preparation method of the α-pyrone mixed terpene dimer Hypericumonates A-C is characterized in that, in step C, Fr.5 is separated by MCI column chromatography, and the concentrations of the methanol / water gradient elution are volume ratios of 60:40, 70:30, 75:25, 80:20, 85:15, 90:10, and 100:0, respectively.

[0017] The method for preparing the α-pyrone mixed terpene dimer Hypericumonates A-C is characterized in that in step E, the concentrations of the gradient elution of petroleum ether / ethyl acetate are volume ratios of 20:1, 10:1, 5:1, and 1:1, respectively; and the semi-preparative high performance liquid chromatography column filler is reversed-phase octadecyl bonded silica gel.

[0018] The application of the α-pyrone mixed heteroterpenoid dimer Hypericumonates AC in the preparation of anti-neuroinflammatory drugs.

[0019] The pharmaceutical composition comprises α-pyrone mixed heteroterpenoid dimer Hypericumonates AC and pharmaceutically acceptable excipients.

[0020] The pharmaceutical composition is characterized in that it contains 0.1-99% by mass of α-pyrone mixed heteroterpenoid dimer Hypericumonates AC, and the rest is a pharmaceutical carrier or excipient.

[0021] Experimental studies have shown that the α-pyrone mixed heteroterpenoid dimer Hypericumonates AC has significant inhibitory activity on NO in mouse microglia (BV-2) induced by lipopolysaccharide (LPS), exhibits significant anti-neuroinflammatory effects, and is more active than the positive control drug minocycline. It can be used to prepare anti-neuroinflammatory drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The structural formula of the α-pyrone mixed heteroterpene dimer compound Hypericumonates A-C of the present invention is:

[0023] Figure 2 This is the nuclear magnetic resonance hydrogen spectrum of the α-pyrone mixed heteroterpene dimer compound Hypericumonate A;

[0024] Figure 3 This is the carbon NMR spectrum of the α-pyrone mixed terpene dimer compound Hypericumonate A;

[0025] Figure 4 This is the nuclear magnetic resonance hydrogen spectrum of Hypericumonate B, a compound of α-pyrone mixed heteroterpene dimer;

[0026] Figure 5 This is the carbon NMR spectrum of Hypericumonate B, a compound of α-pyrone mixed heteroterpene dimer;

[0027] Figure 6 This is the nuclear magnetic resonance hydrogen spectrum of Hypericumonate C, a compound of α-pyrone mixed heteroterpene dimer;

[0028] Figure 7 This is the carbon NMR spectrum of Hypericumonate C, a compound of α-pyrone mixed heteroterpene dimer;

[0029] Figure 8 This is the result of Western blotting experiment of Hypericumonate A, a mixed α-pyrone terpene dimer compound, on LPS-induced BV-2 cells. DETAILED DESCRIPTION

[0030] Example 1

[0031] An α-pyrone mixed heteroterpene dimer compound Hypericumonates A-C, whose structural formula is formula A to C (the numbers in the formula are the positions of carbon atoms in the chemical structure):

[0032]

[0033] The method for preparing the α-pyrone mixed terpenoid dimer Hypericumonates A-C comprises the following steps:

[0034] Step A: taking 50 kg of dried Hypericum perforatum branches and leaves, using 150-250 kg of methanol as solvent, extracting 3-5 times at a temperature of 20-30° C., each extraction time being 4 days, and concentrating under reduced pressure to recover methanol to obtain a crude extract I;

[0035] Step B: using dichloromethane, ethyl acetate and methanol as extraction solvents, the crude extract I obtained in step A was subjected to solid phase extraction to obtain three fractions: component A, component B, and component C; component A was subjected to silica gel column chromatography, and the volume ratio of petroleum ether to dichloromethane was 100:0-0:100. Then, different fractions were detected by thin layer chromatography technology, and seven fractions were obtained by merging according to their thin layer chromatography performance: Fr.1, Fr.2, Fr.3, Fr.4, Fr.5, Fr.6, and Fr.7;

[0036] Step C: fraction Fr.5 was separated by MCI column chromatography, and the eluent was gradient eluted with a volume ratio of methanol to water of 40:60 to 100:0, and the fractions were combined according to the thin layer chromatography performance to obtain 7 sub-fractions: Fr.5.A, Fr.5.B, Fr.5.C, Fr.5.D, Fr.5.E, Fr.5.F and Fr.5.G;

[0037] Step D: Fr.5.D was separated by gel LH-20 column chromatography, eluted with methanol as solvent, and four sub-fractions were obtained: Fr.5.D.1, Fr.5.D.2, Fr.5.D.3 and Fr.5.D.4. Fraction Fr.5.D.2 was further purified by gel column chromatography (methanol) and recrystallization (methanol) to obtain α-pyrone mixed heteroterpenoid dimer Hypericumonate A.

[0038] Step E: Fraction Fr.5.C was subjected to gradient elution by gel column chromatography (methanol) and silica gel column chromatography (petroleum ether and ethyl acetate volume ratio 20:1 to 1:1) to obtain 4 subfractions: Fr.5.C.1, Fr.5.C.2, Fr.5.C.3 and Fr.5.C.4. Component Fr.5.C.3 was further purified by semi-preparative high performance liquid chromatography, with acetonitrile and water volume ratio of 55:45 and flow rate of 3.0 mL / min to obtain α-pyrone mixed heteroterpenoid dimers Hypericumonate B and Hypericumonate C.

[0039] The method for preparing the α-pyrone mixed terpene dimer Hypericumonates A-C is characterized in that in step A, the methanol is 100% industrial methanol; and the extraction method adopted is cold soaking extraction.

[0040] The preparation method of the α-pyrone mixed terpene dimer Hypericumonates A-C is characterized in that, in step B, fraction A is separated by silica gel column chromatography, and the concentrations of the gradient elution of petroleum ether / dichloromethane are volume ratios of 100:0, 80:20, 70:30, 60:40, 50:50, 20:80, and 0:100, respectively.

[0041] The preparation method of the α-pyrone mixed terpene dimer Hypericumonates A-C is characterized in that, in step C, Fr.5 is separated by MCI column chromatography, and the concentrations of the methanol / water gradient elution are volume ratios of 60:40, 70:30, 75:25, 80:20, 85:15, 90:10, and 100:0, respectively.

[0042] The method for preparing the α-pyrone mixed terpene dimer Hypericumonates A-C is characterized in that in step E, the concentrations of the gradient elution of petroleum ether / ethyl acetate are volume ratios of 20:1, 10:1, 5:1, and 1:1, respectively; and the semi-preparative high performance liquid chromatography column filler is reversed-phase octadecyl bonded silica gel.

[0043] The application of the α-pyrone mixed heteroterpenoid dimer Hypericumonates AC in the preparation of anti-neuroinflammatory drugs.

[0044] The pharmaceutical composition comprises α-pyrone mixed heteroterpenoid dimer Hypericumonates AC and pharmaceutically acceptable excipients.

[0045] The pharmaceutical composition is characterized in that it contains 0.1-99% by mass of α-pyrone mixed heteroterpenoid dimer Hypericumonates AC, and the rest is a pharmaceutical carrier or excipient.

[0046] Example 2

[0047] The structural identification of α-pyrone mixed heteroterpene dimer Hypericumonates AC was carried out by high resolution mass spectrometry (HRESIMS), nuclear magnetic resonance spectroscopy ( 1 H NMR, 13 C NMR, 2D-NMR), infrared spectrum (IR), melting point (mp), optical rotation and other data were comprehensively analyzed, and its physical and chemical properties are as follows:

[0048] Hypericumonate A: colorless crystals, (MeOH): mp 193-195°C; IR(KBr)ν max 2973.0, 1709.1, 1638.7, 768.4, and 732.4 cm -1 ; HRESIMS m / z 495.1787[M+Na] + (The calculated value is C 29 H 28 O6Na,495.1778). NMR spectrum 1 H and 13 C NMR data are shown in Table 1.

[0049] Hypericumonate B: colorless powder; IR(KBr)ν max 2974.5,1714.9,1644.5,1582.8,1410.4and 983.8cm -1 ; HRESIMS m / z 461.1926[M+Na] + (The calculated value is C 26 H 30 O6Na,461.1953). NMR spectrum 1 H and 13 C NMR data are shown in Table 1.

[0050] Hypericumonate C: white powder; UV(MeOH)λ max (logε)295.0(3.88)nm; IR(KBr)ν max2967.3,2932.8,1717.7,1645.9,and 989.5cm -1 ; HRESIMS m / z475.2101[M+Na] + (The calculated value is C 27 H 32 O6Na,475.2091). NMR spectrum 1 H and 13 C NMR data are shown in Table 1.

[0051] Table 1: Compounds A to C in deuterated chloroform 1 H and 13 C NMR data (coupling constant J is in Hz, chemical shift δ is in ppm).

[0052]

[0053] Example 3

[0054] In order to further verify the beneficial effects of the compounds of the present invention, the inhibitory effect of Hypericumonates AC prepared in Example 1 on NO in LPS-induced BV-2 cells was used;

[0055] (1) Test samples:

[0056] Preparation of sample solution: Accurately weigh appropriate amount of Hypericumonates A-C and use DMSO to prepare sample solutions of different concentrations for pharmacological activity testing.

[0057] Cell line: Mouse glial cell line BV-2.

[0058] (2) Test method:

[0059] ① MTT method to determine cytotoxicity: Take cells in the logarithmic growth phase, digest and centrifuge, discard the culture medium supernatant, resuspend the cells with 10mL complete culture medium, mix well, take 10μL of cell solution to the counting plate, and count under a microscope. Calculate the required volume of cell solution and culture medium, prepare the cell solution and mix well, then inoculate the cells in a 96-well plate (BV-2 cells: 10000 / well), and add 100μL ddH2O to each well around the 96-well plate. Then put it in a 37℃, 5% CO2 constant temperature incubator. After the cells adhere to the wall and grow for 24 hours, add different concentrations of α-pyrone mixed heteroterpene dimer Hypericumonates AC solution to treat it, so that the final concentration is: 10 or 20μg / mL of the compound. After the cells were treated with drugs and cultured for 24, 48 and 72 hours, 10 μL of MTT was added to each well. After culture at 37°C for 4 hours, the cells were centrifuged at 3000 rpm for 15 minutes, the supernatant of the culture medium was aspirated and discarded, 160 μL of DMSO was added to each well, and the cells were placed on a 37°C constant temperature shaker and shaken at a low speed for 20 minutes to fully dissolve the crystals. The absorbance (OD value) of each well was then measured at OD 490nm using an enzyme-linked immunosorbent assay.

[0060]

[0061] ② Detection of nitric oxide (NO) concentration: Collect cells and plate them in 96-well plates (BV-2: 10,000 cells / well). After 12 hours of cell culture, prepare and add different concentrations of α-pyrone mixed terpene dimer Hypericumonates AC to a final concentration of 10μg / mL or 20μg / mL, as well as positive control minocycline (MINO: 20μg / mL). Incubate for 12 hours, prepare and add LPS to a final concentration of 5μg / mL, and incubate the drug with LPS for 24 hours. Collect the supernatant of the culture medium, centrifuge for 20 minutes, and take the supernatant. Add the prepared standards and samples to a blank 96-well plate at 50μL / well. Add 50μL Griess Reagent I and 50μL Griess Reagent II to each well. The absorbance was measured at a wavelength of 540 nm using an enzyme marker, and a standard curve was drawn with the X-axis as the standard concentration and the Y-axis as the absorbance. The absorbance value was substituted into the regression equation to calculate the concentration of NO in the sample.

[0062] ③IC 50 Test: Same method as above.

[0063] ④Western blot experiment: BV-2 cells were plated in a culture dish (3.6×10 6 / dish), prepared and added different concentrations of the compound Hypericumonate A (5μM, 10μM and 20μM), incubated for 24h, and then added LPS. Prepare protein lysate and add 1% protease inhibitor cocktail and 1% phenylmethylsulfonyl fluoride (PMSF). Protein concentration was detected by BCA protein assay kit. Protein samples were electrophoresed by 10% SDS-PAGE and transferred to polyvinylidene fluoride (PVDF) membrane. The membrane was washed with TBST buffer, blocked in 5% skim milk for 1 hour, and incubated with primary antibody overnight. Finally, protein bands were developed using Odyssey imager.

[0064] (3) Experimental results: According to the above test, at different concentrations (10 or 20 μM), the compounds Hypericumonates A-C showed no cytotoxicity. By measuring the inhibitory effect of Hypericumonates AC on NO in LPS-induced BV-2 cells, the results showed that Hypericumonates AC could significantly inhibit the NO concentration in LPS-induced BV-2 cells, and its IC 50 The value was between 5.88±0.53 and 7.99±1.73. The activity was significantly stronger than that of the positive control drug (minocycline, IC 50 =19.09±1.34); especially the compound Hypericumonate A, IC 50 The results are shown in Table 2. In the Western blot experiment, it can be observed that with the increase of the concentration of the compound Hypericumonate A, the expression of the two proteins iNOS and COX-2 showed a significant downward trend (see Figure 8 , A: Immunoblotting results of iNOS and COX-2 protein expression; B, C: Quantitative analysis using Image J software. Compared with the control group, ### p<0.001; **p<0.01, ***p<0.001 compared with the LPS-induced group).

[0065] Table 2 Inhibitory activity of compounds Hypericumonates A-C on LPS-induced NO concentration in BV-2 cells

[0066]

[0067] (4) Experimental conclusion: The compound Hypericumonates AC has significant inhibitory activity on the NO concentration in LPS-induced BV-2 cells, showing obvious anti-inflammatory effects, and its activity is stronger than that of the positive control drug minocycline. In addition, the compound Hypericumonate A can inhibit the expression of iNOS and COX-2 in a dose-dependent manner. Therefore, the Hypericumonates A-C of the present invention can be used to prepare anti-neuroinflammatory drugs.

[0068] The above is only a preferred embodiment of the present invention. It should be pointed out that for researchers in this technical field, any other improvements and modifications made without departing from the method and content of the present invention should also be regarded as within the scope of protection of the present invention.

Claims

1. An α-pyrone mixed heteroterpenoid dimer Hypericumonates AC, whose structural formula is A to C:

2. A method for preparing the α-pyrone mixed terpenoid dimer Hypericumonates AC as claimed in claim 1, characterized in that: The following steps are involved: Step A: taking 50 kg of dried Hypericum perforatum branches and leaves, using 150-250 kg of methanol as solvent, extracting 3-5 times at a temperature of 20-30° C., each extraction time being 4 days, and concentrating under reduced pressure to recover methanol to obtain a crude extract I; Step B: using dichloromethane, ethyl acetate and methanol as extraction solvents, solid phase extraction is performed on the crude extract I obtained in step A to obtain three fractions: component A, component B, and component C; component A is subjected to silica gel column chromatography, and gradient elution is performed with petroleum ether and dichloromethane in a volume ratio of 100:0-0:100; different fractions are then detected using thin layer chromatography technology, and seven fractions are obtained by merging them according to their thin layer chromatography performance: Fr.1, Fr.2, Fr.3, Fr.4, Fr.5, Fr.6, and Fr.7; Step C: fraction Fr.5 was separated by MCI column chromatography, and the eluent was gradient eluted with a volume ratio of methanol to water of 40:60 to 100:0, and the fractions were combined according to the thin layer chromatography performance to obtain 7 sub-fractions: Fr.5.A, Fr.5.B, Fr.5.C, Fr.5.D, Fr.5.E, Fr.5.F and Fr.5.G; Step D: Fr.5.D was separated by gel LH-20 column chromatography, eluted with methanol as solvent, and four sub-fractions were obtained: Fr.5.D.1, Fr.5.D.2, Fr.5.D.3 and Fr.5.D.

4. Fraction Fr.5.D.2 was further purified by gel column chromatography and recrystallization, and the elution solvent used was methanol, to obtain α-pyrone mixed heteroterpenoid dimer Hypericumonate A; Step E: Fraction Fr.5.C was purified by gel column chromatography and silica gel column chromatography with a gradient elution of petroleum ether and ethyl acetate in a volume ratio of 20:1 to 1:1 to obtain four subfractions: Fr.5.C.1, Fr.5.C.2, Fr.5.C.3 and Fr.5.C.4; component Fr.5.C.3 was further purified by semi-preparative high performance liquid chromatography with an acetonitrile to water volume ratio of 55:45 and a flow rate of 3.0 mL / min to obtain α-pyrone mixed heteroterpene dimers Hypericumonate B and Hypericumonate C.

3. The method for preparing the α-pyrone mixed terpenoid dimer Hypericumonates AC according to claim 2, characterized in that: In step A, the methanol is 100% industrial methanol; and the extraction method is cold soaking extraction.

4. The method for preparing the α-pyrone mixed heteroterpenoid dimer Hypericumonates AC according to claim 2, characterized in that: In step B, fraction A is separated by silica gel column chromatography, and the concentrations of petroleum ether / dichloromethane in gradient elution are volume ratios of 100:0, 80:20, 70:30, 60:40, 50:50, 20:80, and 0:100, respectively.

5. The method for preparing the α-pyrone mixed heteroterpenoid dimer Hypericumonates AC according to claim 2, characterized in that: In step C, Fr.5 is separated by MCI column chromatography, and the concentrations of the gradient elution of methanol / water are volume ratios of 60:40, 70:30, 75:25, 80:20, 85:15, 90:10, and 100:0, respectively.

6. The method for preparing the α-pyrone mixed terpenoid dimer Hypericumonates AC according to claim 2, characterized in that: In step E, the concentrations of the gradient elution of petroleum ether / ethyl acetate are volume ratios of 20:1, 10:1, 5:1, and 1:1, respectively; and the semi-preparative high performance liquid chromatography column filler is reversed-phase octadecyl bonded silica gel.

7. Use of the α-pyrone mixed heteroterpenoid dimer Hypericumonates AC as claimed in claim 1 in the preparation of anti-neuroinflammatory drugs.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the α-pyrone mixed heteroterpenoid dimer Hypericumonates AC according to claim 1 and pharmaceutically acceptable excipients.

9. The pharmaceutical composition according to claim 8, characterized in that The pharmaceutical composition contains 0.1-99% by mass of the α-pyrone mixed heteroterpenoid dimer Hypericumonates A-C according to claim 1, and the rest is a pharmaceutical carrier or excipient.

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