Hypericumonates A–C, a mixed-source α-pyranone heteroterpene dimer, and its preparation method and applications

Abstract

This invention discloses an α-pyranone mixed heteroterpene dimer, Hypericumonates A-C, its preparation method, and its applications, belonging to the field of pharmaceutical chemistry. The α-pyranone mixed heteroterpene dimer, Hypericumonates A-C, has the structure shown in formulas A-C. This class of compounds was isolated for the first time from Hypericum monogynum. The compounds Hypericumonates A-C exhibit significant inhibitory activity against lipopolysaccharide (LPS)-induced NO concentration in BV-2 cells, demonstrating a significant anti-inflammatory effect. Its activity is stronger than that of the positive control drug minocycline. Furthermore, Hypericumonate A can dose-dependently inhibit the expression of iNOS and COX-2, and can be used in the preparation of anti-neuroinflammatory drugs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical chemistry, specifically relating to an α-pyranone mixed-source heteroterpene dimer Hypericumonates AC, its preparation method, and its application. Background Technology

[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 function, promoting nerve repair and maintaining homeostasis; however, long-term neuroinflammation can lead to excessive production of inflammatory mediators, resulting in neurological dysfunction and degeneration, which is associated with various neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, and Huntington's disease. Therefore, the discovery of drugs that inhibit neuroinflammation is of great value and significance. Marketed neuroinflammation treatments, such as memantine and tramiprosate, have unsatisfactory therapeutic effects and significant side effects. Natural medicines, as an important source of innovative drug discovery, have become a popular area for treating neuroinflammation; therefore, the discovery of new anti-neuroinflammation drugs from natural products has attracted the attention of many medicinal chemists. α-Pyranone hybrid heteroterpene dimers are rare natural products formed by a [2+2] cyclization reaction of two α-pyranone dimethylpyran rings via a terminal double bond to an intracyclic double bond or an intracyclic double bond to another intracyclic double bond. They possess unique chemical structures and exhibit excellent anti-neuroinflammatory activity. In this invention, the α-pyranone hybrid heteroterpene dimer Hypericumonates AC is a 6 / 6 / 4-6 / 6 ring system formed by the [2+2] cycloaddition reaction of the terminal double bond of one molecule of α-pyranone dimethylpyran ring to the intracyclic double bond of another molecule of α-pyranone ring, ultimately linked by a C-C bond. Its basic core is a novel, previously unseen skeleton, and it exhibits significant anti-neuroinflammatory activity. Hypericum monogynum, a plant belonging to the genus Hypericum in the family Guttiferae, is rich in α-pyranone mixed heteroterpenoids, among which dimers have not yet been reported in the literature. This plant is widely distributed in southwestern China, and its flowers, roots, and leaves can all be used medicinally, possessing antirheumatic, hepatitis-treating, antibacterial, anti-inflammatory, bronchitis-treating, hemostatic, and tissue-regenerating effects. Current research on the chemical constituents of Hypericum monogynum mainly focuses on phloroglucinols, spironolactones, flavonoids, and triterpenoids. Therefore, discovering novel α-pyranone mixed heteroterpenoid dimers with anti-neuroinflammatory activity from the traditional Chinese medicine Hypericum monogynum has significant research value and importance. Summary of the Invention

[0003] The purpose of this invention is to discover a novel α-pyranone mixed-origin heteroterpene dimer from the traditional Chinese medicine Hypericum perforatum, which possesses anti-neuroinflammatory activity, and its application in the preparation of anti-neuroinflammatory drugs. This invention provides an α-pyranone mixed-origin heteroterpene dimer, its preparation method, and its application. The following technical solution is adopted to achieve the above objective:

[0004] A mixed-source heteroterpene dimer, Hypericumonates A-C, has the structural formulas A-C:

[0005]

[0006] Right now:

[0007]

[0008] The method for preparing the α-pyranone mixed-source heteroterpene dimer Hypericumonates A-C includes the following steps:

[0009] Step A: Take 50 kg of dried Hypericum branches and leaves, use 150-250 kg of methanol as solvent, and extract 3-5 times at a temperature of 20-30℃, with each extraction lasting 4 days. Concentrate under reduced pressure to recover methanol and obtain crude extract I.

[0010] 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: fraction A, fraction B, and fraction C. Fraction A was subjected to silica gel column chromatography with a gradient elution of petroleum ether and dichloromethane at a volume ratio of 100:0 to 0:100. Thin-layer chromatography was then used to detect the different fractions, and they were combined according to their thin-layer chromatographic performance to obtain seven fractions: 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 using a gradient elution of methanol to water at a volume ratio of 40:60 to 100:0. The fractions were combined according to their thin-layer chromatographic characteristics to obtain seven subfractions: 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 column chromatography (LH-20) using methanol as the elution solvent to obtain four sub-fractions: 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 using methanol as the elution solvent to obtain the α-pyranone mixed heteroterpene dimer Hypericumonate A.

[0013] Step E: Fraction Fr.5.C was purified by gel column chromatography and silica gel column chromatography using a gradient elution of petroleum ether and ethyl acetate at 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. Fraction Fr.5.C.3 was further purified by semi-preparative high-performance liquid chromatography at a volume ratio of acetonitrile to water of 55:45 and a flow rate of 3.0 mL / min to obtain the α-pyranone mixed-source heteroterpene dimers Hypericumonate B and Hypericumonate C.

[0014] The method for preparing the α-pyranone mixed-source heteroterpene dimers Hypericumonates A-C is characterized in that, in step A, the methanol is 100% industrial methanol; and the extraction method is cold maceration extraction.

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

[0016] The method for preparing the α-pyranone mixed-source heteroterpene 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 successively 60:40, 70:30, 75:25, 80:20, 85:15, 90:10, and 100:0 (volume ratios).

[0017] The method for preparing the α-pyranone mixed-source heteroterpene dimers Hypericumonates A-C is characterized in that, in step E, the concentrations of the petroleum ether / ethyl acetate gradient elution are successively 20:1, 10:1, 5:1, and 1:1 (volume ratio); and the chromatographic column packing material of the semi-preparative high-performance liquid chromatography is reversed-phase octadecyl bonded silica gel.

[0018] The application of the α-pyranone mixed-source heteroterpene dimer Hypericumonates AC in the preparation of anti-neuroinflammatory drugs.

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

[0020] The pharmaceutical composition is characterized in that it contains 0.1-99% by mass of α-pyranone mixed-source heteroterpene dimers Hypericumonates AC, with the remainder being pharmaceutical carriers or excipients.

[0021] Experimental studies have shown that the α-pyranone mixed heteroterpene dimer Hypericumonates AC exhibits significant inhibitory activity against NO in lipopolysaccharide (LPS)-induced mouse microglia (BV-2), demonstrating a marked anti-neuroinflammatory effect, and its activity is stronger than that of the positive control drug minocycline. It can be used to prepare anti-neuroinflammatory drugs. Attached Figure Description

[0022] Figure 1 The structural formulas of Hypericumonates A-C, a type of α-pyranone mixed-source heteroterpene dimer compound of this invention, are as follows:

[0023] Figure 2 This is the 1H NMR spectrum of Hypericumonate A, a mixed-origin heteroterpene dimer of α-pyranone;

[0024] Figure 3 This is the carbon NMR spectrum of Hypericumonate A, a mixed-origin heteroterpene dimer of α-pyranone;

[0025] Figure 4 This is the 1H NMR spectrum of Hypericumonate B, an α-pyranone mixed-origin heteroterpene dimer.

[0026] Figure 5 This is the carbon NMR spectrum of Hypericumonate B, an α-pyranone mixed-source heteroterpene dimer.

[0027] Figure 6 This is the 1H NMR spectrum of Hypericumonate C, an α-pyranone mixed-origin heteroterpene dimer.

[0028] Figure 7 This is the carbon NMR spectrum of Hypericumonate C, an α-pyranone mixed-origin heteroterpene dimer.

[0029] Figure 8 This is the result of Western blot experiment on LPS-induced BV-2 cells using Hypericumonate A, an α-pyranone mixed-origin heteroterpene dimer. Detailed Implementation

[0030] Example 1

[0031] A type of α-pyranone mixed heteroterpene dimer compound, Hypericumonates A-C, has the structural formula A-C (the numbers in the formula are the carbon atom numbers in the chemical structure):

[0032]

[0033] The method for preparing the α-pyranone mixed-source heteroterpene dimer Hypericumonates A-C includes the following steps:

[0034] Step A: Take 50 kg of dried Hypericum branches and leaves, use 150-250 kg of methanol as solvent, and extract 3-5 times at a temperature of 20-30℃, with each extraction lasting 4 days. Concentrate under reduced pressure to recover methanol and obtain 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: fraction A, fraction B, and fraction C. Fraction A was subjected to silica gel column chromatography with a gradient elution of petroleum ether and dichloromethane at a volume ratio of 100:0 to 0:100. Thin-layer chromatography was then used to detect the different fractions, and they were combined according to their thin-layer chromatographic properties to obtain seven fractions: 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 using a gradient elution of methanol to water at a volume ratio of 40:60 to 100:0. The fractions were combined according to their thin-layer chromatographic characteristics to obtain seven subfractions: 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 column chromatography (LH-20) using methanol as the solvent, yielding four sub-fractions: 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 the α-pyranone mixed heteroterpene dimer Hypericumonate A.

[0038] Step E: Fraction Fr.5.C was eluted by gradient elution using gel column chromatography (methanol) and silica gel column chromatography (with 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. Fraction Fr.5.C.3 was further purified by semi-preparative high-performance liquid chromatography (HPLC) at a volume ratio of acetonitrile to water of 55:45 at a flow rate of 3.0 mL / min to obtain the α-pyranone mixed-source heteroterpene dimers Hypericumonate B and Hypericumonate C.

[0039] The method for preparing the α-pyranone mixed-source heteroterpene dimers Hypericumonates A-C is characterized in that, in step A, the methanol is 100% industrial methanol; and the extraction method used is cold maceration extraction.

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

[0041] The method for preparing the α-pyranone mixed-source heteroterpene 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 successively 60:40, 70:30, 75:25, 80:20, 85:15, 90:10, and 100:0 (volume ratios).

[0042] The method for preparing the α-pyranone mixed-source heteroterpene dimers Hypericumonates A-C is characterized in that, in step E, the concentrations of the petroleum ether / ethyl acetate gradient elution are successively 20:1, 10:1, 5:1, and 1:1 (volume ratio); and the chromatographic column packing material of the semi-preparative high-performance liquid chromatography is reversed-phase octadecyl bonded silica gel.

[0043] The application of the α-pyranone mixed-source heteroterpene dimer Hypericumonates AC in the preparation of anti-neuroinflammatory drugs.

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

[0045] The pharmaceutical composition is characterized in that it contains 0.1-99% by mass of α-pyranone mixed-source heteroterpene dimers Hypericumonates AC, with the remainder being pharmaceutical carriers or excipients.

[0046] Example 2

[0047] Structural identification of the α-pyranone mixed heteroterpene dimer compound Hypericumonates AC: High-resolution mass spectrometry (HRESIMS) and nuclear magnetic resonance spectroscopy were used. 1 H NMR, 13 Based on a comprehensive analysis of data including C NMR, 2D-NMR, infrared spectroscopy (IR), melting point (mp), and optical rotation, its physicochemical properties are as follows:

[0048] Hypericumonate A: Colorless crystals, (MeOH): mp193-195℃; 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). Nuclear magnetic resonance spectrum. 1 H and 13 The 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). Nuclear magnetic resonance spectrum. 1 H and 13 The 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). Nuclear magnetic resonance spectrum. 1 H and 13 The 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 in Hz, chemical shift δ in ppm).

[0052]

[0053] Example 3

[0054] To further verify the beneficial effects of the compounds described in this invention, the inhibitory effect of Hypericumonates AC prepared in Example 1 on NO induced by LPS in BV-2 cells was investigated.

[0055] (1) Test sample:

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

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

[0058] (2) Experimental methods:

[0059] ① MTT assay for cytotoxicity: Cells in logarithmic growth phase were digested and centrifuged, the supernatant was discarded, and the cells were resuspended in 10 mL of complete culture medium. After thorough mixing, 10 μL of the cell solution was transferred to a counting chamber and counted under a microscope. The required volumes of cell solution and culture medium were calculated, and the prepared cell solution was thoroughly mixed. Cells were then seeded in 96-well plates (BV-2 cells: 10,000 / well), and 100 μL of ddH2O was added to each well around the perimeter. The plates were then incubated at 37°C in a 5% CO2 incubator. After 24 hours of cell adhesion and growth, different concentrations of α-pyranone mixed-source heteroterpene dimer Hypericumonates AC solution were added to achieve a final concentration of 10 or 20 μg / mL of the compound. After cells were cultured for 24, 48, and 72 hours following drug treatment, 10 μL of MTT was added to each well. After incubation at 37°C for 4 hours, the cells were centrifuged at 3000 rpm for 15 minutes. The supernatant was then discarded, and 160 μL of DMSO was added to each well. The cells were then placed on a 37°C shaker and shaken at low speed for 20 minutes to fully dissolve the crystals. The absorbance (OD value) of each well was then measured at OD 490 nm using an ELISA reader.

[0060]

[0061] ② Nitric oxide (NO) concentration detection: Cells were collected and seeded in 96-well plates (BV-2: 10,000 cells / well). After culturing the cells for 12 hours, different concentrations of α-pyranone mixed-origin heteroterpene dimers (Hypericumonates AC) were prepared and added to a final concentration of 10 μg / mL or 20 μg / mL, along with a positive control (minocycline: 20 μg / mL). After incubation for 12 hours, LPS was prepared and added to a final concentration of 5 μg / mL. The drug and LPS were co-incubated for 24 hours. The culture supernatant was collected, centrifuged for 20 minutes, and the supernatant was collected. The prepared standards and samples were added to blank 96-well plates at 50 μL / well. 50 μL of Griess Reagent I and 50 μL of Griess Reagent II were added to each well, respectively. The absorbance was measured at 540 nm using an ELISA reader. A standard curve was plotted with the X-axis representing the standard concentration and the Y-axis representing the absorbance. The absorbance values ​​were then substituted into the regression equation to calculate the NO concentration in the sample.

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

[0063] ④Western blot experiment: BV-2 cells were seeded in culture dishes (3.6×10⁻⁶). 6Protein samples were prepared by adding different concentrations of Hypericumonate A (5 μM, 10 μM, and 20 μM) to a sample / plate and incubating for 24 h, followed by the addition of LPS. Protein lysis buffer was prepared and mixed with a 1% protease inhibitor mixture and 1% benzyl sulfonyl fluoride (PMSF). Protein concentration was determined using a BCA protein assay kit. Protein samples were subjected to 10% SDS-PAGE electrophoresis and transferred to a polyvinylidene fluoride (PVDF) membrane. The membrane was washed with TBST buffer, blocked in 5% skim milk for 1 h, and incubated with primary antibody overnight. Finally, protein bands were visualized using an Odyssey imager.

[0064] (3) Experimental Results: According to the above experiments, compounds Hypericumonates A-C did not exhibit cytotoxicity at different concentrations (10 or 20 μM). The inhibitory effect of Hypericumonates AC on LPS-induced NO in BV-2 cells was determined. The results showed that Hypericumonates AC significantly inhibited LPS-induced NO concentration in BV-2 cells, with an IC50 value of [missing information]. 50 The values ​​ranged from 5.88±0.53 to 7.99±1.73. The activity was significantly stronger than the positive control (minocycline, IC50). 50 =19.09±1.34); especially compound Hypericumonate A, IC 50 The value was 5.88 ± 0.53, as shown in Table 2 below. In the Western blot experiment, it was observed that with increasing concentrations of the compound Hypericumonate A, the expression of both proteins, iNOS and COX-2, showed a significant downregulation trend (see Table 2). Figure 8 A: Immunoblotting results of iNOS and COX-2 protein expression; B, C: Quantitative analysis performed using ImageJ software. Compared with the control group, ### p<0.001; compared with the LPS-induced group, **p<0.01, ***p<0.001).

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

[0066]

[0067] (4) Experimental Conclusions: Compound Hypericumonates AC significantly inhibited NO concentration in LPS-induced BV-2 cells, exhibiting a marked anti-inflammatory effect, and its activity was stronger than that of the positive control drug minocycline. Furthermore, compound Hypericumonate A dose-dependently inhibited the expression of iNOS and COX-2. Therefore, Hypericumonates A-C of the present invention can be used to prepare anti-neuroinflammatory drugs.

[0068] The above description is only a preferred embodiment of the present invention. It should be noted that any other improvements and modifications made by researchers in this field without departing from the method and content of the present invention should also be considered within the scope of protection of the present invention.

Claims

1. A kind α -Pyranone mixed heteroterpene dimers Hypericumonates A–C, with the structural formula A ~ C:

2. A device as described in claim 1 α A method for preparing hypericumonates A–C, a mixed-source heteroterpene dimer of pyranones, characterized in that... Includes the following steps: Step A: Take 50 kg of dried Hypericum branches and leaves, use 150–250 kg of methanol as solvent, extract 3–5 times at 20–30℃, each extraction time is 4 days, concentrate under reduced pressure to recover methanol to obtain crude extract I. 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: fraction A, fraction B, and fraction C. Fraction A was subjected to silica gel column chromatography with a gradient elution of petroleum ether and dichloromethane at a volume ratio of 100:0−0:

100. Thin-layer chromatography was then used to detect the different fractions, and they were combined according to their thin-layer chromatographic performance to obtain seven fractions: 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 using a gradient elution of methanol to water at a volume ratio of 60:40 to 100:

0. The fractions were combined according to their thin-layer chromatographic characteristics to obtain seven subfractions: 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 column chromatography (GLC) using an LH-20 column, eluting with methanol to obtain four sub-fractions: 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 GLC and recrystallization, using methanol as the eluting solvent. α - Hypericumonate A, a mixed-origin heteroterpene dimer of pyranones; Step E: Fraction Fr.5.C was purified by gel column chromatography and silica gel column chromatography. The gel column was eluted with methanol, and the silica gel column was eluted with a gradient of petroleum ether and ethyl acetate at a volume ratio of 20:1 to 1:1, yielding four subfractions: Fr.5.C.1, Fr.5.C.2, Fr.5.C.3, and Fr.5.C.

4. Fraction Fr.5.C.3 was further purified by semi-preparative high-performance liquid chromatography (HPLC) at a volume ratio of acetonitrile to water of 55:45 and a flow rate of 3.0 mL / min. α -Pyranone mixed-origin heteroterpene dimers Hypericumonate B and Hypericumonate C.

3. The method according to claim 2 α A method for preparing hypericumonates A–C, a mixed-source heteroterpene dimer of pyranones, characterized in that... In step A, the methanol is 100% industrial methanol; the extraction method is cold maceration extraction.

4. The method according to claim 2 α A method for preparing hypericumonates A–C, a mixed-source heteroterpene dimer of pyranones, characterized in that... In step B, fraction A is separated by silica gel column chromatography, with the concentrations of petroleum ether / dichloromethane eluted in the following volume ratios: 100:0, 80:20, 70:30, 60:40, 50:50, 20:80, and 0:

100.

5. The method according to claim 2 α A method for preparing hypericumonates A–C, a mixed-source heteroterpene dimer of pyranones, characterized in that... In step C, Fr.5 was separated using MCI column chromatography, with methanol / water gradient elution concentrations of 60:40, 70:30, 75:25, 80:20, 85:15, 90:10, and 100:0, respectively.

6. The method according to claim 2 α A method for preparing hypericumonates A–C, a mixed-source heteroterpene dimer of pyranones, characterized in that... In step E, the concentrations of the petroleum ether / ethyl acetate gradient elution are successively 20:1, 10:1, 5:1, and 1:1 (volume ratio); the column packing material for the semi-preparative high-performance liquid chromatography is reversed-phase octadecyl bonded silica gel.

7. A device as described in claim 1 α Application of Hypericumonates A–C, a mixed-origin heteroterpene dimer of pyranones, in the preparation of anti-neuroinflammatory drugs.

8. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the product of claim 1. α - Hypericumonates A–C 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 ingredient described in claim 1. α - Hypericumonates A–C, a mixed-origin heteroterpenoid dimer, with the remainder being pharmaceutically acceptable excipients.

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