CN114044734B - Rosin alkane diterpene and preparation method and application thereof - Google Patents
Rosin alkane diterpene and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to the technical field of separation and purification of schizonepeta makino, in particular to abietyl diterpene and a preparation method and application thereof. The invention discloses a compound 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dimethyl-1-methylen-henanthrene-2, 9-diol (nepetabrate D) for the first time, and experiments prove that the compound has stronger inhibition effect on RAW 264.7 cells, staphylococcus aureus and escherichia coli by performing in vitro anti-inflammatory and antibacterial pharmacodynamics experiments, so that the compound can be used for preparing anti-inflammatory and antibacterial drugs or/and health care products for preventing and treating inflammation and antibacterial.
Description
Technical Field
The invention relates to the technical field of separation and purification of schizonepeta majora, in particular to abietyl diterpene, and a preparation method and application thereof, wherein abietyl diterpene is 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dimethyl-1-methyl-phenanthrene-2, 9-diol (nepetabrate D) for short.
Background
Herba Schizonepetae Nepeta bracteata Benth is a Nepeta plant of Labiatae, and is mainly distributed in the countries such as Pakistan, nepal, iran, etc., and is named as "ancestor hair", known as Shenxiang Xiang, and has wide clinical application mainly imported. The whole herb is used as a medicine, has faint scent of qi, light taste and slight dampness, enters lung and liver channels, has the effects of relieving cough and asthma, clearing heat and promoting diuresis, and is clinically used for treating symptoms such as tracheitis, cough and asthma, cold fever, difficult urination and the like. In addition, the schizonepeta herb has the characteristics of easy source, low cost, reliable clinical curative effect, small toxic and side effects and the like. So far, the focus of research on schizonepeta makino and basic theoretical research are not seen, and modern pharmacology shows that the extract has remarkable anti-inflammatory and antibacterial activities, but related research on chemical components is not carried out yet, and the schizonepeta makino is probably a good species for treating respiratory diseases, so that the schizonepeta makino has been paid attention to in recent years.
The drug effect of the schizonepeta majora is mainly derived from terpenoid compounds, including monoterpenes and diterpenoid compounds, wherein the diterpenoid components of the dominant component have good anti-inflammatory and antibacterial activities, so that the diterpenoid monomer compounds of the schizonepeta majora are developed and utilized, the potential medicinal value of the schizonepeta majora is further excavated, the structure and physical and chemical properties of the monomer compounds of the schizonepeta majora are determined and characterized, and the schizonepeta majora has important significance in development and utilization of the schizonepeta majora.
Disclosure of Invention
The invention provides 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dimethyl-1-methylen-henapherene-2, 9-diol (nepetabrate D) and a preparation method and application thereof, overcomes the defects of the prior art, and firstly discloses 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dimethyl-1-methylen-henapherene-2, 9-diol (nepetabrate D) and application thereof in preparing anti-inflammatory and antibacterial drugs or/and preparing anti-inflammatory and antibacterial health care products.
One of the technical schemes of the invention is realized by the following measures: 1,2,3, 4a,9,10 a-octah ydro-7-isopropyyl-2, 4 a-dimethyl-1-methylethylehenapherene-2, 9-diol (nepetabrate D), the chemical structural formula is
The following are further optimizations and/or improvements to one of the above-described inventive solutions:
the preparation method comprises the following steps: crushing schizonepeta makino, adding ethanol, soaking for 3-4 hours at room temperature, heating and reflux-extracting for 3 times at 50-60 ℃ for 1-3 hours each time, combining the reflux extracting solutions each time, decompressing and recovering and concentrating to obtain schizonepeta makino total extractum; dispersing the total extract of schizonepeta makino with water, and extracting with petroleum ether and dichloromethane in sequence to obtain a petroleum ether part and a dichloromethane part; step three, taking petroleum ether part extractum, and obtaining 8 fractions after gradient elution and separation by a silica gel column chromatography, wherein the gradient elution liquid of the silica gel column chromatography comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is sequentially 1:0, 100:1 and 50: 1. 25:1, 8:1, 5:1, 1:1, 1:0; fourthly, eluting and separating the 7 th fraction in the 8 fractions by a silica gel column chromatography gradient to obtain 5 fractions, wherein the silica gel column chromatography gradient eluent comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 15:1, 5:1 and 3: 1. 1:1, 0:1; fifthly, separating the 3 rd fraction in the 5 fractions by silica gel column chromatography gradient elution to obtain 3 fractions, wherein the silica gel column chromatography gradient elution comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 4:1, 2:1 and 1:1, a step of; and sixthly, eluting, purifying and separating the 3 rd fraction in the 3 obtained fractions by using a high performance liquid chromatography gradient, collecting the eluate, and obtaining 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-meth ylenephenanthrene-2,9-diol (nepetabrate D) at 25.6 minutes.
In the first step, 8ml to 10ml of ethanol is added to each 1g of schizonepeta.
In the sixth step, the eluent of the high performance liquid chromatography gradient elution is a mixed solution of methanol and water, wherein the volume ratio of the methanol to the water is 85:15.
the second technical scheme of the invention is realized by the following measures: the preparation method of 1,2,3, 4a,9,10 a-octah ydro-7-isopropyl-2,4 a-dimethyl-1-methylen-henaphenthrene-2, 9-diol (nepetabrate D) comprises the following steps: crushing schizonepeta makino, adding ethanol, soaking for 3-4 hours at room temperature, heating and reflux-extracting for 3 times at 50-60 ℃ for 1-3 hours each time, combining the reflux extracting solutions each time, decompressing and recovering and concentrating to obtain schizonepeta makino total extractum; dispersing the total extract of schizonepeta makino with water, and extracting with petroleum ether and dichloromethane in sequence to obtain a petroleum ether part and a dichloromethane part; step three, taking petroleum ether part extractum, and obtaining 8 fractions after gradient elution and separation by a silica gel column chromatography, wherein the gradient elution liquid of the silica gel column chromatography comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is sequentially 1:0, 100:1 and 50: 1. 25:1, 8:1, 5:1, 1:1, 1:0; fourthly, eluting and separating the 7 th fraction in the 8 fractions by a silica gel column chromatography gradient to obtain 5 fractions, wherein the silica gel column chromatography gradient eluent comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 15:1, 5:1 and 3: 1. 1:1, 0:1; fifthly, separating the 3 rd fraction in the 5 fractions by silica gel column chromatography gradient elution to obtain 3 fractions, wherein the silica gel column chromatography gradient elution comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 4:1, 2:1 and 1:1, a step of; and sixthly, eluting, purifying and separating the 3 rd fraction in the 3 obtained fractions by using a high performance liquid chromatography gradient, collecting the eluate, and obtaining 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-me thylenephenanthrene-2,9-diol (nepetabrate D) at 25.6 minutes.
The following is a further optimization and/or improvement of the second technical scheme of the invention:
in the first step, 8ml to 10ml of ethanol is added to each 1g of schizonepeta.
In the sixth step, the eluent of the high performance liquid chromatography gradient elution is a mixed solution of methanol and water, wherein the volume ratio of the methanol to the water is 85:15.
the third technical scheme of the invention is realized by the following measures: application of 1,2,3, 4a,9,10 a-octah ydro-7-isopropyl-2,4 a-dimethyl-1-methylen-henaphenthrene-2, 9-diol (nepetabrate D) in preparing medicines for preventing inflammation and inhibiting bacteria.
The fourth technical scheme of the invention is realized by the following measures: application of 1,2,3, 4a,9,10 a-octah ydro-7-isopropyl-2,4 a-dimethyl-1-methylen-henaphenthrene-2, 9-diol (nepetabrate D) in preparing anti-inflammatory and antibacterial drugs.
The fifth technical scheme of the invention is realized by the following measures: application of 1,2,3, 4a,9,10 a-octah ydro-7-isopropyl-2,4 a-dimethyl-1-methylen-henaphenthrene-2, 9-diol (nepetabrate D) in preparing health care products for preventing and treating inflammation and inhibiting bacteria.
The invention discloses a compound 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dime-1-methyl-phenanthrene-2, 9-diol (nepetabrate D) for the first time, and the compound is subjected to in vitro anti-inflammatory and antibacterial pharmacodynamics experiments, so that the experiment shows that the compound has stronger inhibition effect on RAW 264.7 cells, staphylococcus aureus and escherichia coli, and the compound can be used for preparing anti-inflammatory and antibacterial drugs or/and preparing health care products for preventing and treating inflammation and antibacterial.
Drawings
FIG. 1 is a chemical structure diagram of 1,2,3, 4a,9,10 a-octahydro-7-isopopyl-2, 4 a-dimethyl-1-methylethylehenanthrene-2, 9-diol (nepetabrate D) according to the present invention.
FIG. 2 is a schematic diagram of 1,2,3, 4a,9,10 a-octahydro-7-isopopyl-2, 4 a-dimethyl-1-methylethylehenanthrene-2, 9-diol (nepetabrate D) of the present invention 1 H-NMR spectrum.
FIG. 3 is a schematic diagram of 1,2,3, 4a,9,10 a-octahydro-7-isopopyl-2, 4 a-dimethyl-1-methylethylehenanthrene-2, 9-diol (nepetabrate D) of the present invention 13 C-APT spectrogram.
Detailed Description
The present invention is not limited by the following examples, and specific embodiments can be determined according to the technical scheme and practical situations of the present invention. The various chemical reagents and chemical supplies mentioned in the invention are all commonly known and used in the prior art unless specified otherwise; the percentages in the invention are mass percentages unless specified otherwise; the solutions in the invention are aqueous solutions in which the solvent is water unless otherwise specified, for example, the hydrochloric acid solution is hydrochloric acid aqueous solution; the room temperature and the room temperature in the present invention generally refer to temperatures ranging from 15 ℃ to 25 ℃, and are generally defined as 25 ℃.
The invention is further described below with reference to examples:
example 1: the 1,2,3, 4a,9,10 a-octahydro-7-isopropyyl-2, 4a-dimethyl-1-met hylenephenanthrene-2,9-diol (nepetabrate D) is characterized by the chemical structural formula
Subjecting 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dime-thyl-1-methylen henapherene-2, 9-diol (nepetabrate D) described in example 1 of the present invention to nuclear magnetic resonance hydrogen spectroscopy 1 H-NMR) and nuclear magnetic resonance carbon spectrum 13 C-APT).
1,2,3, 4a,9,10 a-octahydro-7-isopropyyl-2, 4a-dimethyl-1-m ethylenephenanthrene-2,9-diol (nepetabrate D) of this example 1 The H-NMR spectrum is shown in FIG. 2As shown.
The 1,2,3, 4a,9,10 a-octahydro-7-isopopyl-2, 4 a-dimethyl-1-methylethylehenanthene-2, 9-diol (nepetabrate D) described in this example 13 The C-APT spectrum is shown in FIG. 3.
The peaks shown in FIG. 2 and FIG. 3 were assigned to the peaks shown in Table 1, and the data shown in Table 1 shows that 1,2,3, 4a,9,10 a-octahydro-7-isopopyl-2, 4 a-dimethyl-1-methylethylehenapherene-2, 9-diol (nepetabrate D) of the present example has a chemical structure shown in FIG. 1, and is easily dissolved in chloroform or methanol.
Example 2: the 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-met hylenephenanthrene-2,9-diol (nepetabrate D) is prepared according to the following method: crushing schizonepeta makino, adding ethanol, soaking for 3-4 hours at room temperature, heating and reflux-extracting for 3 times at 50-60 ℃ for 1-3 hours each time, combining the reflux extracting solutions each time, decompressing and recovering and concentrating to obtain schizonepeta makino total extractum; dispersing the total extract of schizonepeta makino with water, and extracting with petroleum ether and dichloromethane in sequence to obtain a petroleum ether part and a dichloromethane part; step three, taking petroleum ether part extractum, and obtaining 8 fractions after gradient elution and separation by a silica gel column chromatography, wherein the gradient elution liquid of the silica gel column chromatography comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is sequentially 1:0, 100:1 and 50: 1. 25:1, 8:1, 5:1, 1:1, 1:0; fourthly, eluting and separating the 7 th fraction in the 8 fractions by a silica gel column chromatography gradient to obtain 5 fractions, wherein the silica gel column chromatography gradient eluent comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 15:1, 5:1 and 3: 1. 1:1, 0:1; fifthly, separating the 3 rd fraction in the 5 fractions by silica gel column chromatography gradient elution to obtain 3 fractions, wherein the silica gel column chromatography gradient elution comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 4:1, 2:1 and 1:1, a step of; and sixthly, eluting, purifying and separating the 3 rd fraction in the 3 obtained fractions by a high performance liquid chromatography gradient, and collecting the eluate to obtain 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dimethyl-1-methylenanthrene-2, 9-diol (nepetabrate D) at 25.6 minutes.
Example 3: as an optimization of the above examples, the 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dimethyl-1-methylethylehenapherene-2, 9-diol (nepetabrate D) was prepared as follows: firstly, crushing schizonepeta makino, adding ethanol, soaking for 3 hours or 4 hours at room temperature, heating and reflux-extracting for 3 times at 50 ℃ or 60 ℃ for 1 hour or 3 hours each time, combining reflux extracting solutions each time, decompressing and recovering and concentrating to obtain schizonepeta makino total extractum; dispersing the total extract of schizonepeta makino with water, and extracting with petroleum ether and dichloromethane in sequence to obtain a petroleum ether part and a dichloromethane part; step three, taking petroleum ether part extractum, and obtaining 8 fractions after gradient elution and separation by a silica gel column chromatography, wherein the gradient elution liquid of the silica gel column chromatography comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is sequentially 1:0, 100:1 and 50: 1. 25:1, 8:1, 5:1, 1:1, 1:0; fourthly, eluting and separating the 7 th fraction in the 8 fractions by a silica gel column chromatography gradient to obtain 5 fractions, wherein the silica gel column chromatography gradient eluent comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is sequentially 1:1, 5:1 and 3: 1. 1:1, 0:1; fifthly, separating the 3 rd fraction in the 5 fractions by silica gel column chromatography gradient elution to obtain 3 fractions, wherein the silica gel column chromatography gradient elution comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 4:1, 2:1 and 1:1, a step of; and sixthly, eluting, purifying and separating the 3 rd fraction in the 3 obtained fractions by a high performance liquid chromatography gradient, and collecting the eluate to obtain 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dimethyl-1-methyl-henanthrene-2, 9-diol (nepetabrate D) at 25.6 minutes.
Example 4: as an optimization of the above examples, in the first step, 8ml to 10ml of ethanol was added per 1g of schizonepeta makino.
Example 5: as an optimization of the above embodiment, in the sixth step, the eluent of the high performance liquid chromatography gradient elution is a mixed solution of methanol and water, wherein the volume ratio of methanol to water is 85:15.
example 6: the application of the 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-met hylenephenanthrene-2,9-diol (nepetabrate D) in preparing medicaments for preventing inflammation and inhibiting bacteria.
Example 7: the 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-met hylenephenanthrene-2,9-diol (nepetabrate D) is used for preparing anti-inflammatory and antibacterial drugs.
Example 8: the 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-met hylenephenanthrene-2,9-diol (nepetabrate D) is applied to the preparation of health care products for preventing and treating inflammation and bacteriostasis.
Example 9: the 1,2,3, 4a,9,10 a-octahydro-7-isopopyl-2, 4a-dimethyl-1-met hylenephenanthrene-2,9-diol (nepetabrate D) is obtained according to the following method: crushing schizonepeta makino, adding ethanol, soaking for 3 hours at room temperature, heating and reflux-extracting for 3 times at 50 ℃ for 2 hours each time, combining the reflux extracting solutions each time, decompressing and recovering and concentrating to obtain schizonepeta makino total extractum; dispersing the total extract of schizonepeta makino with water, and extracting with petroleum ether and dichloromethane in sequence to obtain a petroleum ether part and a dichloromethane part; step three, taking petroleum ether part extractum, and obtaining 8 fractions after gradient elution and separation by a silica gel column chromatography, wherein the gradient elution liquid of the silica gel column chromatography comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is sequentially 1:0, 100:1 and 50: 1. 25:1, 8:1, 5:1, 1:1, 1:0; fourthly, eluting and separating the 7 th fraction in the 8 fractions by a silica gel column chromatography gradient to obtain 5 fractions, wherein the silica gel column chromatography gradient eluent comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 15:1, 5:1 and 3: 1. 1:1, 0:1; fifthly, separating the 3 rd fraction in the 5 fractions by silica gel column chromatography gradient elution to obtain 3 fractions, wherein the silica gel column chromatography gradient elution comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 4:1, 2:1 and 1:1, a step of; and sixthly, eluting, purifying and separating the 3 rd fraction in the 3 obtained fractions by a high performance liquid chromatography gradient, and collecting the eluate to obtain 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dimethyl-1-methyl-henanthrene-2, 9-diol (nepetabrate D) at 25.6 minutes.
The 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dime-thyl-1-methylen henapherene-2, 9-diol (nepetabrate D) obtained in example 9 of the present invention was subjected to in vitro anti-inflammatory and antibacterial pharmacodynamic experiments, which utilized MTT colorimetry.
Taking 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephen anthrene-2,9-diol (nepetabrate D) as experimental group, aspin (aspirin, anti-inflammatory drug) as control group, setting blank group, selecting RAW 264.7 cells as experimental group, and diluting culture medium, and taking 6×10 cells 4 The density of each ml is inoculated in a 96-well plate, 100 mu l of each well is cultured normally in an incubator for 24 hours, and the corresponding medicines are added into each group, so that the final concentration of each group of medicines is respectively 2.5 mu g/ml (1 group), 5 mu g/ml (2 group), 10 mu g/ml (3 group), 20 mu g/ml (4 group) and 40 mu g/ml (5 group), 5 concentrations are set, and 3 compound wells are arranged for each concentration; after 48 hours of incubation, 10. Mu.l of MTT was added to each well for staining; after further culturing for four hours, the stock culture was aspirated, 150. Mu.l of DMSO was added to each well, the mixture was allowed to oscillate on a shaking table at low speed for 10min to allow the crystals to be sufficiently dissolved, and the optical density value was measured at 570nm wavelength of an ELISA, and 50% Inhibitory Concentration (IC) was calculated from the optical density value 50 Mu M), optical density value calculating IC 50 The calculation method is a prior known technology. IC of experimental group and control group on RAW 264.7 cells 50 As shown in table 2. As can be seen from the data in Table 2, 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dimethyl-1-methyethylhenapherene-2, 9-diol (nepetabrate D) of the present invention has a certain inhibition effect on RAW 264.7 cells.
Taking 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephen anthrene-2,9-diol (nepetabrate D) as experimental group, penicillin (amoxicillin, antibacterial agent) as control group, and setting up blank groupThe test group, the control group and the blank group select staphylococcus aureus and escherichia coli as test objects, samples are diluted by a certain multiple by sterile water or DMSO, 100 mu L of samples are centrifugally taken and added into holes of an indicator bacteria plate to be used as antibacterial test holes, fresh cultured indicator strains are taken, the concentration of bacterial suspension is regulated to 0.5McFarland and is uniformly coated on the plate, the plates are placed for culturing for 24 hours or 48 hours at corresponding temperature, and the colony growth condition of each plate is observed. 100 μl of fresh medium was added to the wells as negative control wells and amoxicillin solution as positive control. After dilution of the medium, the culture medium was diluted to 6X 10 4 The density of each ml is inoculated in a 96-well plate, 100 mu l of each well is cultured normally in an incubator for 24 hours, and the corresponding medicines are added into each group, so that the final concentration of each group of medicines is respectively 2.5 mu g/ml (1 group), 5 mu g/ml (2 group), 10 mu g/ml (3 group), 20 mu g/ml (4 group) and 40 mu g/ml (5 group), 5 concentrations are set, and 3 compound wells are arranged for each concentration; after 48 hours of incubation, 10. Mu.l of MTT was added to each well for staining; after further culturing for four hours, the stock culture was aspirated, 150. Mu.l of DMSO was added to each well, the mixture was allowed to oscillate on a shaking table at low speed for 10min to allow the crystals to be sufficiently dissolved, and the optical density value was measured at 570nm wavelength of an ELISA, and 50% Inhibitory Concentration (IC) was calculated from the optical density value 50 Mu M), optical density value calculating IC 50 The calculation method is a prior known technology. IC of experiment group, control group against staphylococcus aureus and escherichia coli 50 As shown in table 3. As can be seen from the data in Table 3, the 1,2,3, 4a,9,10 a-octah ydro-7-isopropyl-2,4 a-dimethyl-1-methyethylhenapherene-2, 9-diol (nepetabrate D) of the invention has certain inhibition effect on staphylococcus aureus and escherichia coli.
In summary, the invention discloses the compounds 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4 a-dimethyl-1-methylen-2, 9-diol (nepetabrate D) for the first time, and the compounds are subjected to in vitro anti-inflammatory and antibacterial pharmacodynamic experiments, so that the experiments show that the compounds have stronger inhibition effects on RAW 264.7 cells, staphylococcus aureus and escherichia coli, and the compounds can be used for preparing anti-inflammatory and antibacterial drugs or/and antibacterial health care products.
The technical characteristics form the embodiment of the invention, have stronger adaptability and implementation effect, and can increase or decrease unnecessary technical characteristics according to actual needs so as to meet the requirements of different situations.
TABLE 1
TABLE 2
TABLE 3 Table 3
Claims (7)
1. 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephenanthrene-2,9-diol, characterized in that the 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephenanthrene-2,9-diol is nepetabrate D, and has the chemical structural formula of
2. A process for the preparation of 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephenanthrene-2,9-diol according to claim 1, characterized in that it is carried out according to the following steps: crushing schizonepeta makino, adding ethanol, soaking for 3-4 hours at room temperature, heating and reflux-extracting for 3 times at 50-60 ℃ for 1-3 hours each time, combining the reflux extracting solutions each time, decompressing and recovering and concentrating to obtain schizonepeta makino total extractum; dispersing the total extract of schizonepeta makino with water, and extracting with petroleum ether and dichloromethane in sequence to obtain a petroleum ether part and a dichloromethane part; step three, taking petroleum ether part extractum, and obtaining 8 fractions after gradient elution and separation by a silica gel column chromatography, wherein the gradient elution liquid of the silica gel column chromatography comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is sequentially 1:0, 100:1 and 50: 1. 25:1, 8:1, 5:1, 1:1, 1:0; fourthly, eluting and separating the 7 th fraction in the 8 fractions by a silica gel column chromatography gradient to obtain 5 fractions, wherein the silica gel column chromatography gradient eluent comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 15:1, 5:1 and 3: 1. 1:1, 0:1; fifthly, separating the 3 rd fraction in the 5 fractions by silica gel column chromatography gradient elution to obtain 3 fractions, wherein the silica gel column chromatography gradient elution comprises petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 4:1, 2:1 and 1:1, a step of; and sixthly, eluting, purifying and separating the 3 rd fraction in the 3 obtained fractions by high performance liquid chromatography gradient, collecting the eluate, and obtaining nepetabrate D at 25.6 minutes, namely the 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephenanthrene-2, 9-dihydric alcohol.
3. The process for the preparation of 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephenanthrene-2,9-diol according to claim 2, characterized in that in the first step 8ml to 10ml of ethanol is added per 1g of schizonepeta megalobum.
4. A process for the preparation of 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephenanthrene-2,9-diol according to claim 2 or 3, characterized in that in the sixth step, the eluent of high performance liquid chromatography gradient elution is a mixture of methanol and water, wherein the volume ratio of methanol to water is 85:15.
5. use of 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephenanthrene-2,9-diol according to claim 1 for the preparation of a medicament for preventing inflammation and inhibiting bacteria.
6. Use of 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephenanthrene-2,9-diol according to claim 1 for the preparation of anti-inflammatory and antibacterial agents.
7. Use of 1,2,3, 4a,9,10 a-octahydro-7-isopropyl-2,4a-dimethyl-1-methylenephenanthrene-2,9-diol according to claim 1 for the preparation of a health product for preventing and treating inflammation and inhibiting bacteria.
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