Diterpenoid component in hyssopus harzianum ethanol extract as well as extraction method and application of diterpenoid component

Abstract

The invention relates to the technical field of hyssop officinalis extracts, in particular to a diterpenoid component in a hyssop officinalis ethanol extract as well as an extraction method and application of the diterpenoid component, the diterpenoid component is obtained at the 23rd minute by adding ethanol into the overground part of hyssop officinalis, extracting, concentrating, eluting and separating the obtained petroleum ether part extract and purifying and separating the third fraction. The invention discloses a diterpenoid component in the hyssop officinalis ethanol extract for the first time, namely a compound (1R, 5R, 6 'R, 7a' R)-6, 6, 7 ', 7'-tetramethyl tetrahydrospiro [bicyclo [3.1. 1] heptane-2, 2 '-[6, 7a] methylbenzofuran]-3, 4' (5 'H)-diketone, and in-vitro anti-tumor pharmacodynamic experiments find that the diterpenoid component has a relatively strong inhibition effect on HepG-2 cells and HeLa cells, and can be used for preparing a medicine for treating tumors. Therefore, the compound can be used for preparing medicines for preventing liver cancer or / and cervical cancer and medicines for resisting liver cancer or / and cervical cancer.

Description

Technical Field

[0001] The invention relates to the technical field of hyssopus officinalis extracts, in particular to a diterpenoid component in a hyssopus officinalis ethanol extract, an extraction method and application thereof, and also relates to application of the diterpenoid component in the hyssopus officinalis ethanol extract in preparing tumor prevention or / and anti-tumor drugs. The diterpenoid component in the hyssopus officinalis ethanol extract is a compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione. Background Art

[0002] Terpenoids are a large class of natural products, and are also one of the most complex and diverse natural product categories in terms of structure. Currently, terpenoids can be isolated from plants of the genus Hyssop.

[0003] Hyssopus officinalis is a plant of the genus Hyssopus in the family Lamiaceae. "Annotation to Medical Classics": "It can relieve cough and resolve phlegm, relieve inflammation and relieve asthma, make the skin bright, eliminate wounds, soften and reduce swelling, eliminate tinnitus, activate blood circulation and remove blood stasis, and expel intestinal worms. It is mainly used to treat pleurisy, pneumonia, asthma, whooping cough, pale complexion, facial wounds, hard inflammation and swelling, tinnitus, blood stasis in the white of the eyes, and intestinal worms." "Bai Di Yi Pharmacy": "It can relieve inflammation and relieve asthma, relieve cough and resolve phlegm, dispel cold and relieve pain, clear the liver and soften the spleen, promote diuresis and reduce swelling, eliminate the poison of poisonous insects, and expel intestinal worms. It is mainly used to treat pneumonia, shortness of breath and asthma, long-term cough, cold, toothache, sciatica, back pain, liver obstruction and spleen swelling, edema, and poisonous insect bites. It can be seen that the whole plant of hard-pointed Hyssopus officinalis can be used as medicine, which has the effects of clearing heat and detoxification and anti-inflammatory. At the same time, modern pharmacological studies have shown that the terpenoid compounds it contains have a wide range of pharmacological activities including central nervous system effects, cardiovascular system effects, anti-tumor, anti-virus, etc.

[0004] The Chinese patent application document with the authorization announcement number CN115010589B discloses a kind of abietic acid diterpenoid component in Hyssopus officinalis extract and its preparation method and application, wherein the abietic acid diterpenoid component in Hyssopus officinalis extract is the abbreviation of Nepetabrate E. However, it only involves that Nepetabrate E has a strong anti-inflammatory effect on RAW264.7 cells stimulated by lipopolysaccharide (LPS), so that Nepetabrate E can be used as a drug for the preparation of anti-inflammatory drugs and anti-inflammatory drugs.

[0005] The Chinese patent application document with publication number CN115010604A discloses a diterpene component in Hyssopus officinalis and its preparation method and application, wherein the diterpene component in Hyssopus officinalis is the abbreviation of Nepetabrate I. Animal experiments show that the Nepetabrate I of the present invention has significant anti-inflammatory and anti-inflammatory effects, can play a synergistic anti-inflammatory role, has a definite efficacy, high safety, and easy-to-obtain raw materials, and can be used in the preparation of drugs with anti-inflammatory effects. However, it only involves the application of Nepetabrate I in the preparation of drugs with anti-inflammatory effects.

[0006] Therefore, developing and utilizing the terpenoid compounds of Hyssopus sclerotium, further exploring its potential medicinal value, determining and characterizing the structure and physicochemical properties of its monomer compounds, and systematically evaluating the anti-tumor ability of terpenoid compounds are of great significance for the development and utilization of Hyssopus sclerotium plants. Summary of the invention

[0007] The invention provides a diterpenoid component in a Herba Hyssopus sclerotii ethanol extract and an extraction method and application thereof, which overcome the shortcomings of the above-mentioned prior art and disclose for the first time a compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione, which has a strong inhibitory effect on HepG-2 cells and HeLa cells, so that the compound can be used for preparing a drug for preventing liver cancer and / or cervical cancer and a drug for resisting liver cancer and / or cervical cancer.

[0008] One of the technical solutions of the present invention is achieved by the following measures: a compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione, having a chemical structural formula of .

[0009] The following is a further optimization and / or improvement of one of the above-mentioned technical solutions: The above was extracted according to the following method: The first step is to grind and sieve the aerial part of the hard-pointed Hyssopus officinalis and add ethanol, soak it at room temperature, then heat it under reflux extraction, combine each reflux extract, recover it under reduced pressure, and concentrate it to obtain the hard-pointed Hyssopus officinalis total extract; The second step is to disperse the total extract of Hyssopus officinalis into a suspension with water, extract it with petroleum ether, dichloromethane and ethyl acetate in sequence, and concentrate the extract to obtain extracts of various parts respectively; The third step is to obtain 8 fractions by gradient elution of the petroleum ether extract from a silica gel column. In the fourth step, the third fraction among the eight fractions obtained was purified and separated by high performance liquid chromatography gradient elution, and the eluate was collected to obtain the compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione at 23 minutes.

[0010] In the first step above, 8 mL to 12 mL of ethanol was added per 1 g of the aerial part of Hyssopus officinalis.

[0011] In the first step, the soaking time is 4.5 hours to 5.5 hours, the heating temperature is 40° C. to 60° C., the reflux extraction times are 3 times, and the reflux extraction time each time is 2 hours to 3 hours.

[0012] In the third step, the gradient eluent of the silica gel column chromatography includes chloroform and methanol, and the volume ratios of chloroform and methanol are 1:0, 100:1, 40:1, 20:1, 10:1, 5:1, 2:1 and 1:1, respectively.

[0013] In the fourth step, the eluent for the HPLC gradient elution is a mixture of methanol and water, wherein the volume ratio of methanol to water is 75:25.

[0014] The second technical solution of the present invention is achieved by the following measures: A method for extracting a compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione is carried out according to the following method: The first step is to grind and sieve the aerial part of the hard-pointed Hyssopus officinalis and add ethanol, soak it at room temperature, then heat it under reflux extraction, combine each reflux extract, recover it under reduced pressure, and concentrate it to obtain the hard-pointed Hyssopus officinalis total extract; The second step is to disperse the total extract of Hyssopus officinalis into a suspension with water, extract it with petroleum ether, dichloromethane and ethyl acetate in sequence, and concentrate the extract to obtain extracts of various parts respectively; The third step is to take the petroleum ether extract and separate it by silica gel column chromatography with gradient elution to obtain 8 fractions; In the fourth step, the third fraction among the eight fractions obtained was purified and separated by high performance liquid chromatography gradient elution, and the eluate was collected to obtain the compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione at 23 minutes.

[0015] The following is a further optimization and / or improvement of the second technical solution of the above invention: In the first step above, 8 mL to 12 mL of ethanol was added per 1 g of the aerial part of Hyssopus officinalis.

[0016] In the first step, the soaking time is 4.5 hours to 5.5 hours, the heating temperature is 40° C. to 60° C., the reflux extraction times are 3 times, and the reflux extraction time each time is 2 hours to 3 hours.

[0017] In the third step, the gradient eluent of the silica gel column chromatography includes chloroform and methanol, and the volume ratios of chloroform and methanol are 1:0, 100:1, 40:1, 20:1, 10:1, 5:1, 2:1 and 1:1, respectively.

[0018] In the fourth step, the eluent for the HPLC gradient elution is a mixture of methanol and water, wherein the volume ratio of methanol to water is 75:25.

[0019] The third technical solution of the present invention is achieved by the following measures: use of a compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione in the preparation of drugs for preventing liver cancer and / or cervical cancer.

[0020] The fourth technical solution of the present invention is achieved by the following measures: use of a compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione in the preparation of anti-liver cancer and / or cervical cancer drugs.

[0021] The present invention discloses the compound for the first time, and through in vitro anti-tumor pharmacodynamic experiments, finds that it has a strong inhibitory effect on HepG-2 cells and HeLa cells, so that it can be used as a drug for preventing liver cancer and / or cervical cancer and a drug for resisting liver cancer and / or cervical cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Attached Figure 1 Hyssopusone E of the present invention 1 H-NMR spectrum.

[0023] Attached Figure 2 Hyssopusone E of the present invention 13 C-APT spectrum.

[0024] Attached Figure 3 Hyssopusone E of the present invention1 H- 1 HCOSY spectrum.

[0025] Attached Figure 4 It is the HSQC spectrum of Hyssopusone E of the present invention.

[0026] Attached Figure 5 It is the HMBC spectrum of Hyssopusone E of the present invention.

[0027] Attached Figure 6 It is the NOESY spectrum of Hyssopusone E of the present invention. DETAILED DESCRIPTION

[0028] The present invention is not limited by the following embodiments, and the specific implementation method can be determined according to the technical scheme of the present invention and the actual situation. Unless otherwise specified, the various chemical reagents and chemicals mentioned in the present invention are all chemical reagents and chemicals known and used in the prior art; unless otherwise specified, the percentages in the present invention are all mass percentages; unless otherwise specified, the solutions in the present invention are all aqueous solutions with water as the solvent, for example, hydrochloric acid solution is a hydrochloric acid aqueous solution; the normal temperature and room temperature in the present invention generally refer to a temperature of 15°C to 25°C, and are generally defined as 25°C.

[0029] The present invention will be further described below in conjunction with embodiments: Example 1: The compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione has a chemical structural formula of .

[0030] The present invention discloses for the first time a diterpenoid component in the ethanol extract of Hyssopus sclerotium, namely the compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione, with a systematic name of (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methanobenzofuran]-3,4'(5'H)-dione (also referred to as hyssopusone E). Experiments show that the compound has a strong inhibitory effect on HepG-2 cells and He La cells, thereby making Hyssopusone E can be used for preparing tumor prevention (liver cancer and / or cervical cancer) drugs and anti-tumor (liver cancer and / or cervical cancer) drugs.

[0031] Example 2: As an optimization of the above example, the following method is used to extract: The first step is to grind and sieve the aerial part of the hard-pointed Hyssopus officinalis and add ethanol, soak it at room temperature, then heat it under reflux extraction, combine each reflux extract, recover it under reduced pressure, and concentrate it to obtain the hard-pointed Hyssopus officinalis total extract; The second step is to disperse the total extract of Hyssopus officinalis into a suspension with water, extract it with petroleum ether, dichloromethane and ethyl acetate in sequence, and concentrate the extract to obtain extracts of various parts respectively; The third step is to take the petroleum ether extract and separate it by silica gel column chromatography with gradient elution to obtain 8 fractions; In the fourth step, the third fraction among the eight fractions obtained was purified and separated by high performance liquid chromatography gradient elution, and the eluate was collected to obtain the compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione at 23 minutes.

[0032] Example 3: As an optimization of the above example, in the first step, 8 mL to 12 mL of ethanol is added to every 1 g of the aerial part of Hyssopus officinalis.

[0033] Example 4: As an optimization of the above example, in the first step, the soaking time is 4.5 hours to 5.5 hours, the heating temperature is 40°C to 60°C, the reflux extraction times is 3 times, and the reflux extraction time each time is 2 hours to 3 hours.

[0034] Example 5: As an optimization of the above example, in the third step, the silica gel column chromatography gradient eluent includes chloroform and methanol, and the volume ratios of chloroform and methanol are 1:0, 100:1, 40:1, 20:1, 10:1, 5:1, 2:1, and 1:1, respectively.

[0035] Example 6: As an optimization of the above example, in the fourth step, the eluent for the HPLC gradient elution is a mixture of methanol and water, wherein the volume ratio of methanol to water is 75:25.

[0036] Example 7: The extraction method of the compound 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione is carried out according to the following method: The first step is to grind and sieve the aerial part of the hard-pointed Hyssopus officinalis and add ethanol, soak it at room temperature, then heat it under reflux extraction, combine each reflux extract, recover it under reduced pressure, and concentrate it to obtain the hard-pointed Hyssopus officinalis total extract; The second step is to disperse the total extract of Hyssopus officinalis into a suspension with water, extract it with petroleum ether, dichloromethane and ethyl acetate in sequence, and concentrate the extract to obtain extracts of various parts respectively; The third step is to take the petroleum ether extract and separate it by silica gel column chromatography with gradient elution to obtain 8 fractions; In the fourth step, the third fraction among the eight fractions obtained was purified and separated by high performance liquid chromatography gradient elution, and the eluate was collected to obtain the compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione at 23 minutes.

[0037] Example 8: Use of the compound 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione in the preparation of drugs for preventing liver cancer and / or cervical cancer.

[0038] Example 9: Use of the compound 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione in the preparation of drugs for treating liver cancer and / or cervical cancer.

[0039] Example 10: The compound 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione was extracted according to the following method: The first step is to grind and sieve 6 kg of the aerial part of hard-pointed Hyssopus officinalis and add 40 L of ethanol, soak at room temperature for 3 hours, heat to 50°C, and reflux extract 3 times, each time for 2 hours, combine the reflux extracts of each time, recover under reduced pressure, and concentrate to obtain the total extract of hard-pointed Hyssopus officinalis; In the second step, 437.0 g of the total extract of Hyssopus sibiricum was dispersed into a suspension with water, and then extracted with petroleum ether, dichloromethane and ethyl acetate in sequence, and the extract was concentrated to obtain extracts of various parts; The third step is to take 134.8 g of petroleum ether extract and separate it by gradient elution on a silica gel column chromatography to obtain 8 fractions, wherein the volume ratios of chloroform and methanol in the gradient eluent of the silica gel column chromatography are 1:0, 100:1, 40:1, 20:1, 10:1, 5:1, 2:1, and 1:1, respectively; In the fourth step, the third fraction among the eight fractions obtained was purified and separated by high performance liquid chromatography gradient elution, and the eluate was collected to obtain the compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione at 23 minutes.

[0040] The compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione obtained in Example 10 of the present invention was subjected to nuclear magnetic resonance hydrogen spectrum ( 1 H-NMR) and carbon nuclear magnetic resonance spectroscopy ( 13 C-APT) analysis. 1 H-NMR spectrum, 13 C-APT spectrum, 1 H- 1 HCOSY spectrum, HSQC spectrum, HMBC spectrum, NOESY spectrum such as Figures 1 to 6 shown.

[0041] Combination Figures 1 to 6 , perform graph analysis, Figure 1 and Figure 2 Each peak is assigned Figure 1 and Figure 2 The peak attributions are shown in Table 1.

[0042] pass Figure 3 , Figure 4 From the data in Table 1, it can be seen that the chemical structure of the compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione of the present invention is as follows: .

[0043] The compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione in the present invention is an amorphous powder and is easily soluble in chloroform and methanol.

[0044] The compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione of the present invention was subjected to an in vitro antitumor pharmacodynamics experiment using the MTT colorimetric method.

[0045] The compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione was used as the experimental group, and Paclitaxel (paclitaxel, an anti-tumor drug) was used as the control group. HepG-2 (human liver cancer cell) cells and HeLa (human cervical cancer) cells were selected as experimental objects in the experimental group and the control group. After the culture medium was diluted, 6×10 4 / mL was inoculated in a 96-well plate, with 100μl in each well. After normal culture in the incubator for 24 hours, the corresponding drugs were added to each group, so that the final concentrations of the drugs in each group were 2.5μg / mL (group 1), 5μg / mL (group 2), 10μg / mL (group 3), 20μg / mL (group 4), and 40μg / mL (group 5), respectively. There were 5 concentrations in total, and 3 replicates for each concentration; after culture for 48 hours, MTT10μL was added to each well for staining; after further culture for four hours, the original culture medium was aspirated and discarded, DMSO150μL was added to each well, and the wells were shaken at a low speed on a shaker for 10 minutes to fully dissolve the crystals, and the optical density value was detected at a wavelength of 570nm on an enzyme-linked immunosorbent assay, and the 50% inhibition concentration (IC) was calculated based on the optical density value. 50 , μg / mL), optical density value was used to calculate IC 50 The calculation method of IC of HepG-2 cells and HeLa cells in the experimental group and the control group is known in the art. 50 As shown in Table 2.

[0046] It can be seen from the data in Table 2 that the compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione described in the present invention has a certain inhibitory effect on both HepG-2 cells and HeLa cells.

[0047] In summary, the present invention discloses the compound for the first time, and through in vitro antitumor pharmacodynamic experiments, it is found that the compound has a strong inhibitory effect on HepG-2 cells and HeLa cells, thereby enabling it to be used as a drug for preventing liver cancer and / or cervical cancer and a drug for resisting liver cancer and / or cervical cancer.

[0048] The above technical features constitute the embodiments of the present invention, which have strong adaptability and implementation effect. Non-essential technical features can be added or reduced according to actual needs to meet the requirements of different situations.

Claims

1. A compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione, characterized in that The chemical structure is 。 2. The compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione according to claim 1, characterized in that Extracted as follows: The first step is to grind and sieve the aerial part of the hard-pointed Hyssopus officinalis and add ethanol, soak it at room temperature, then heat it under reflux extraction, combine each reflux extract, recover it under reduced pressure, and concentrate it to obtain the hard-pointed Hyssopus officinalis total extract; The second step is to disperse the total extract of Hyssopus officinalis into a suspension with water, extract it with petroleum ether, dichloromethane and ethyl acetate in sequence, and concentrate the extract to obtain extracts of various parts respectively; The third step is to take the petroleum ether extract and separate it by silica gel column chromatography with gradient elution to obtain 8 fractions; In the fourth step, the third fraction among the eight fractions obtained was purified and separated by high performance liquid chromatography gradient elution, and the eluate was collected to obtain the compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione at 23 minutes.

3. The compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione according to claim 2, characterized in that In the first step, 8 mL to 12 mL of ethanol was added per 1 g of the aerial part of Hyssopus officinalis.

4. The compound according to claim 1, 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione, characterized in that In the first step, the soaking time is 4.5 hours to 5.5 hours, the heating temperature is 40° C. to 60° C., the reflux extraction times is 3 times, and the reflux extraction time each time is 2 hours to 3 hours.

5. The compound 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione according to claim 1, characterized in that In the third step, the silica gel column chromatography gradient eluent includes chloroform and methanol, and the volume ratios of chloroform and methanol are 1:0, 100:1, 40:1, 20:1, 10:1, 5:1, 2:1, and 1:1, respectively.

6. The compound according to claim 5, 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione, characterized in that In the fourth step, the eluent for the HPLC gradient elution is a mixture of methanol and water, wherein the volume ratio of methanol to water is 75:

25.

7. A method for extracting the compound 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione according to claim 1, characterized in that: Proceed as follows: The first step is to grind and sieve the aerial part of the hard-pointed Hyssopus officinalis and add ethanol, soak it at room temperature, then heat it under reflux extraction, combine each reflux extract, recover it under reduced pressure, and concentrate it to obtain the hard-pointed Hyssopus officinalis total extract; The second step is to disperse the total extract of Hyssopus officinalis into a suspension with water, extract it with petroleum ether, dichloromethane and ethyl acetate in sequence, and concentrate the extract to obtain extracts of various parts respectively; The third step is to take the petroleum ether extract and separate it by silica gel column chromatography with gradient elution to obtain 8 fractions; In the fourth step, the third fraction among the eight fractions obtained was purified and separated by high performance liquid chromatography gradient elution, and the eluate was collected to obtain the compound (1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione at 23 minutes.

8. The method for extracting the compound 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione according to claim 7, characterized in that: In the first step, 8 mL to 12 mL of ethanol is added to every 1 g of the aerial part of Hyssopus officinalis; or / and, in the first step, the soaking time is 4.5 hours to 5.5 hours, the heating temperature is 40°C to 60°C, the reflux extraction is performed 3 times, and the reflux extraction time each time is 2 hours to 3 hours; or / and, in the third step, the silica gel column chromatography gradient eluent includes chloroform and methanol, and the volume ratio of chloroform to methanol is 1:0, 100:1, 40:1, 20:1, 10:1, 5:1, 2:1, and 1:1, respectively; or / and, in the fourth step, the eluent of the high performance liquid chromatography gradient elution is a mixture of methanol and water, wherein the volume ratio of methanol to water is 75:

25.

9. Use of the compound 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione according to any one of claims 1 to 6 in the preparation of a drug for preventing liver cancer and / or cervical cancer.

10. Use of the compound 1R, 5R, 6'R, 7a'R)-6,6,7',7'-tetramethyltetrahydrospiro[bicyclo[3.1.1]heptane-2,2'-[6,7a]methylbenzofuran]-3,4'(5'H)-dione according to any one of claims 1 to 6 in the preparation of drugs for treating liver cancer or / and cervical cancer.

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