CN116970017B - Tetracyclic triterpene compound and preparation method and application thereof - Google Patents
Tetracyclic triterpene compound and preparation method and application thereof Download PDFInfo
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- -1 triterpene compound Chemical class 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 150000001875 compounds Chemical class 0.000 claims abstract description 45
- 241000414067 Inonotus obliquus Species 0.000 claims abstract description 21
- 230000003110 anti-inflammatory effect Effects 0.000 claims abstract description 9
- 230000002757 inflammatory effect Effects 0.000 claims abstract description 9
- 239000003814 drug Substances 0.000 claims abstract description 8
- 230000014509 gene expression Effects 0.000 claims abstract description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 51
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 30
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 20
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 14
- 238000010828 elution Methods 0.000 claims description 14
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 10
- 235000019253 formic acid Nutrition 0.000 claims description 10
- 239000000243 solution Substances 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 239000002024 ethyl acetate extract Substances 0.000 claims description 8
- 239000003208 petroleum Substances 0.000 claims description 8
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- HYKJOXSCXUCBNL-UHFFFAOYSA-N chloroform ethyl acetate methanol toluene Chemical compound CO.C(C)(=O)OCC.ClC(Cl)Cl.C1(=CC=CC=C1)C HYKJOXSCXUCBNL-UHFFFAOYSA-N 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims 1
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- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
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- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Rheumatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Pain & Pain Management (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Steroid Compounds (AREA)
Abstract
The invention belongs to the technical field of natural medicines, and particularly relates to a tetracyclic triterpene compound in inonotus obliquus, and a preparation method and application thereof. In vitro anti-inflammatory activity experiments show that the compound has remarkable inhibition effect on inflammatory factor NO, and can effectively inhibit the gene expression level of the inflammatory mediators increased in RAW264.7 cells. The invention provides an alternative compound for developing new anti-inflammatory medicaments, which has very important significance for comprehensive development and utilization of inonotus obliquus.
Description
Technical Field
The invention relates to the technical field of natural products and medicines, in particular to a tetracyclic triterpene compound extracted from inonotus obliquus, a preparation method and application thereof.
Background
Inonotus obliquus (Inonotus obliquus) is a fungus for both medicine and food of the genus Fuscoporia (Fuscoporia) of the order Hymenochaetales (Hymenochaetaceae). Inonotus obliquus can be used for preventing and treating heart disease, diabetes and various cancers (gastric cancer, liver cancer, intestinal cancer, etc.). Research on inonotus obliquus phytochemistry shows that the chemical components of the inonotus obliquus phytochemistry mainly comprise lanolate alkane type in tetracyclic triterpene compounds, and also comprise a small amount of pentacyclic triterpene compounds, steroid compounds, saccharide compounds, phenols and the like. Modern pharmacological researches show that inonotus obliquus has good effects of regulating blood sugar, resisting tumor, resisting inflammation and resisting oxidation, and also has effects of resisting platelet aggregation and lowering blood pressure.
Therefore, the novel compound with medicinal value is extracted, separated and developed from natural inonotus obliquus, and has great significance for comprehensive development and utilization of inonotus obliquus.
Disclosure of Invention
In view of the above, the invention provides a tetracyclic triterpene compound separated and purified from inonotus obliquus, and a preparation method and application thereof.
The invention provides a novel tetracyclic triterpene compound, which is obtained by separating and purifying Fuscoporia obliqua, wherein the structural formula of the tetracyclic triterpene compound is shown as the formula (I):
the invention also provides a preparation method of the tetracyclic triterpene compound, which comprises the following steps:
s1, weighing and drying inonotus obliquus, crushing, leaching with ethanol, and concentrating the extracting solution under reduced pressure to obtain ethanol extract;
s2, dissolving the ethanol extract with methanol, then adding petroleum ether for extraction, and concentrating the methanol part under reduced pressure to obtain a methanol extract;
s3, dissolving the methanol extract again by using a 9:1 (v/v) water and methanol mixed solution, then adding ethyl acetate for extraction, and concentrating an ethyl acetate part under reduced pressure to obtain an ethyl acetate extract;
s4, performing normal phase silica gel column chromatography on the ethyl acetate extract, performing gradient elution, detecting and combining similar components by TLC to obtain (A-N) 14 components;
s5, performing medium-pressure reversed phase column chromatography on the obtained K components, performing gradient elution, detecting and combining similar components by TLC, and finally obtaining (K-1-K-14) 14 components;
s6, separating the component K-8 through semi-preparative high performance liquid chromatography to obtain the compound shown in the formula (I).
Further, in step S4, conditions for gradient elution of the ethyl acetate extract are as follows: gradient elution was performed with petroleum ether and ethyl acetate at a volume ratio of 20:1, 10:1, 9:1, 8:2, 7:3, 6:4, 1:1, 0:1.
Further, in step S5, the conditions for gradient elution of component K are: gradient elution was performed with water and methanol at volumes of 90:10, 70:30, 50:50, 30:70, 0:100.
Further, in step S6, the high performance liquid chromatography conditions of component K-8 are: isocratic elution is carried out by utilizing acetonitrile and formic acid aqueous solution according to the volume ratio of the acetonitrile to the formic acid aqueous solution of 40:60 at the flow rate of 3 mL/min; wherein the volume percentage of formic acid in the formic acid aqueous solution is 0.1 percent.
The tetracyclic triterpene compound can be applied to the preparation of anti-inflammatory drugs.
The technical scheme provided by the invention has the beneficial effects that: the invention obtains 1 new compound by separating and purifying the ethanol extract of medicinal fungus inonotus obliquus, comprehensively applies various spectrum analysis means and determines that the compound is lanolate alkane type tetracyclic triterpene compound; through in vitro anti-inflammatory activity experiments on the obtained compound, the compound is found to have remarkable inhibition effect on the release of inflammatory factor NO and the expression level of inflammatory mediators IL-1 beta, IL-6, iNOS and COX-2 mRNA; the invention provides an alternative compound for developing new anti-inflammatory medicaments, and has very important significance for comprehensive development and utilization of inonotus obliquus.
Drawings
FIG. 1 is a flow chart of extraction and separation of tetracyclic triterpene compounds prepared in example 1 of the invention;
FIG. 2 shows a compound obtained in example 1 of the present invention 1 H-NMR(500MHz,CD 3 OD) profile;
FIG. 3 shows a compound obtained in example 1 of the present invention 13 C-NMR(125MHz,CD 3 OD) profile;
fig. 4 is a graph of the DEPT (Distortionless Enhancement by Polarization Transfer, distortion free polarization transfer technique) (θ=90°) of the compound prepared in example 1 of the present invention;
fig. 5 is a DEPT (θ=135°) spectrum of the compound prepared in example 1 of the present invention;
FIG. 6 is a graph showing HSQC (Heteronuclear Singular Qantum Correlation, heteronuclear single quantum relationship) of the compound produced in example 1 of the present invention;
FIG. 7 is a chart showing the HMBC (heteronuclear multi-carbon correlation spectrum of 1H detected heteronuclear multiple bond correlation,1H) spectrum of the compound produced in example 1 of the present invention;
FIG. 8 shows a compound obtained in example 1 of the present invention 1 H- 1 H COSY (Correlation spectroscopy, correlation spectrum) profile;
FIG. 9 is a ROESY (Rotating Frame Overhauser Effect Spectroscopy, rotating coordinate System NOE spectrum) spectrum of the compound produced in example 1 of the present invention;
FIG. 10 is a UV spectrum of a compound prepared in example 1 of the present invention;
FIG. 11 is a spectrum of HR-ESI-MS of the compound according to the present invention prepared in example 1;
FIG. 12 is the effect of varying concentrations of the compound of example 1 on RAW264.7 cell viability;
FIG. 13 is the effect of the compound of example 1 on NO release, wherein CON: a control group; LPS: a model group; statistical data are expressed as mean ± SEM, ### p<number 0.001, # represents control vs model group; * P<0.001，**p<0.01，*p<0.05, number represents model group vs dosing group;
FIG. 14 is the effect of the compound of example 1 on inflammatory mediator mRNA expression levels, wherein CON: a control group; LPS: a model group; DEX: dexamethasone; statistical data are expressed as mean ± SEM, ### p<number 0.001, # represents control vs model group; * P<0.001，**p<0.01，*p<Number 0.05 indicates model group vs dosing group.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be further described with reference to specific examples.
The experimental methods used in the following examples are conventional methods unless otherwise specified.
Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1
The embodiment provides a preparation process for extracting tetracyclic triterpene compounds from inonotus obliquus, which comprises the following specific steps:
step S1, weighing 10.25kg of dried inonotus obliquus, crushing, leaching with 95% (v/v) ethanol for 3 times, soaking for 24 hours at room temperature each time, carrying out suction filtration after each time of soaking, combining the extracting solutions obtained after 3 times of suction filtration, and concentrating under reduced pressure to obtain a total extract (170 g); inonotus obliquus used in this example was purchased from Yichang City, hubei province, and identified as Inonotus obliquus (Inonotus obliquus) of genus Fuscoporia of the order Phanerochaete, class of Phanerochaete, university of south China, professor Liu Xinqiao.
S2, dissolving the total extract with methanol, then adding petroleum ether for extraction, and concentrating a methanol part under reduced pressure to obtain a methanol extract (160 g);
step S3, redissolving the methanol extract by using a 9:1 (v/v) water and methanol mixed solution, then adding ethyl acetate for extraction, and concentrating an ethyl acetate part under reduced pressure to obtain an ethyl acetate extract (80 g).
Step S4, carrying out normal phase silica gel column chromatography on the ethyl acetate extract, carrying out gradient elution on petroleum ether and ethyl acetate (petroleum ether: ethyl acetate=20:1, 10:1, 9:1, 8:2, 7:3, 6:4, 1:1, 0:1, v/v), detecting and combining similar components by TLC (developing agent is chloroform: toluene: ethyl acetate: methanol=10:4:1 (0.5-1.5), v/v; the dosage of methanol increases along with the increase of the polarity of the compound), obtaining 14 components, sequentially marking the components as A-N according to the polarity, collecting eluent with the volume ratio of petroleum ether to ethyl acetate of 6:4, numbering as K, and concentrating under reduced pressure until the eluent is dried for later use.
Step S5, loading K components (3.50 g) by a dry method, performing gradient elution by using methanol and water (water: methanol=90:10, 70:30, 50:50, 30:70, 0:100, v/v), detecting and combining similar components by TLC (thin-layer chromatography, thin-layer chromatography, developing agent is chloroform: toluene: ethyl acetate: methanol=10:4:1 (0.5-1.5), v/v, the dosage of the methanol increases along with the increase of the polarity of the compound), finally obtaining 14 components, sequentially marking the components as K-1-K-14 according to the polarity from small to large, collecting eluent with the volume ratio of water: methanol of 50:50, numbering K-8, concentrating to dryness under reduced pressure for later use.
Step S6, purifying the obtained K-8 fraction (58.9 mg) by HPC (High Performance iquid Chromatography, high performance liquid chromatography) using se:Sup>A semi-preparative column YMC-Pack ODS-A (250×10mm,5 μm), isocratically eluting with acetonitrile and an aqueous formic acid solution (0.1% by volume of formic acid in the aqueous formic acid solution, the balance being water) at se:Sup>A flow rate of 3 m/min at se:Sup>A volume ratio of 40:60, and taking se:Sup>A retention time t R The substance=45.36 min, which is the tetracyclic triterpene compound of the invention, has a weight of 2.14mg.
The flow chart of the above steps is shown in fig. 1.
The compound prepared in this example was subjected to high resolution mass spectrometry, ultraviolet spectrometry, optical rotation, nuclear magnetic resonance analysis, thereby determining the structure of the compound.
Physicochemical data and spectroscopic data for the compounds are as follows:
a pale yellow amorphous powder; HR-ESI-MS m/z 473.36273[ M+H ]] + (calcd for C 30 H 49 O 4 ,473.36254)；(c 0.05,MeOH)；UV(MeOH)λ max (log ε) 250 (3.67); the nuclear magnetic resonance data of the compounds are shown in Table (2). 1 H NMR(500MHz,CD 3 OD) spectrum is shown in figure 2, 13 C-NMR(125MHz,CD 3 OD) spectrum is shown in fig. 3, dept (θ=90°) spectrum is shown in fig. 4, dept (θ=135°) spectrum is shown in fig. 5, hsqc spectrum is shown in fig. 6, hmbc spectrum is shown in fig. 7, 1 H- 1 the H COSY spectrum is shown in FIG. 8, the ROESY spectrum is shown in FIG. 9, the UV spectrum is shown in FIG. 10, and the HR-ESI-MS spectrum is shown in FIG. 11.
Table 1: chinese and english naming of compounds
Table 2: NMR data of the compound (Record in CD 3 OD)
Measured at a) CD 3 OD
Example 2: in vitro anti-inflammatory Activity test
In vitro anti-inflammatory activity experiments were performed on the compound prepared in example 1:
materials and reagents: lipopolysaccharide (LPS), dexamethasone, and dimethyl sulfoxide were purchased from Sigma, usa; high sugar DMEM media was purchased from Gibco company, usa; penicillin-streptomycin solution was purchased from GE company usa; fetal bovine serum was purchased from GE company in the united states; CCK-8 kit was purchased from Biosharp, guangzhou; nitric oxide detection kits were purchased from Shanghai Biyun biotechnology limited.
Test tumor cell line: macrophage RAW264.7, available from Wohunorace life technologies Co., ltd (Cat NO: CL-0190).
CCK-8 detection of cell viability
The experimental method comprises the following steps:
RAW264.7 cells were cultured in DMEM high-sugar medium (penicillin, streptomycin diabody) containing 10% fetal bovine serum to about 80% passage, and RAW264.7 cells (1×10) 4 Well) was inoculated into a 96-well plate with a final volume of 100. Mu.L, and cultured in a cell culture incubator at 37℃for 12 hours. The compound of example 1 was prepared in DMSO at a stock concentration of 10mM, diluted to different concentrations (50, 100. Mu.M) in DMEM medium, and added to the 96-well plate at a final volume of 200. Mu.L. The experiments were run with both blank (no cells, DMEM medium only) and control (DMEM medium with cells). After 24h incubation in the incubator, 10. Mu.L of CCK-8 solution was added, incubation in the incubator was continued for 1h protected from light, and then OD values of each well were determined at a wavelength of 450 nm. Cell viability was calculated (see FIG. 12) as calculated by (OD Experimental group -OD Blank group )/(OD Control group -OD Blank group )×100％。
Conclusion of experiment:
from fig. 12, it is clear that the cytotoxicity screening result shows that the compound has no significant toxic effect on cells at 50, 100 μm concentration.
(II) determination of NO Activity
The experimental method comprises the following steps:
RAW264.7 cells were cultured in DMEM high-sugar medium (penicillin, streptomycin diabodies) containing 10% fetal bovine serum to about 80% passage, and then 1X 10 cells were cultured 4 RAW264.7 cells per well were seeded in 96-well plates at 5% co 2 Culturing in a cell culture box at 37 ℃ for 6 hours. Different concentrations (5, 10, 20, 40. Mu.M) of the compound of example 1 and 1. Mu.g/mL of Lipopolysaccharide (LPS) were prepared in DMEM medium. The old medium was discarded from the wells, the blank was only added with DMEM medium (without cells), the control was added with DMEM medium (with cells), the model was added with 1. Mu.g/mL of LPS, the dosing was added with 1. Mu.g/mL of LPS and the compound of example 1 at different concentrations, 100. Mu.L of each well was added and incubated in an incubator for 12h. Then 50. Mu.L of supernatant was measured for the content of nitric oxide in the supernatant by Griess method to determine IC 50 Values, experimental results are shown in FIG. 13.
Conclusion of experiment:
the concentrations of anti-inflammatory activity were set to 5, 10, 20, 40. Mu.M by CCK-8 experiments. The release of NO was significantly increased in the model group after stimulation of inflammatory cells with LPS compared to the control group. Meanwhile, the decrease in the amount of NO released with the increase in the concentration of the compound compared with the model group indicates that the compound of the present invention has an inhibitory effect on NO, and IC is calculated 50 The value was 36.7. Mu.M.
(III) determination of anti-inflammatory Activity
The experimental method comprises the following steps:
RAW264.7 cells were cultured in DMEM high-sugar medium (penicillin, streptomycin diabody) containing 10% fetal bovine serum to about 80% passage, and RAW264.7 macrophages (3×10) 4 individual/mL) was inoculated in 12-well plates at 5% co 2 After incubation for 24h in a 37℃cell incubator, the compound of example 1 was added at different concentrations (10, 20, 40. Mu.M) and incubation was continued for 1h before addition1. Mu.g/mL lipopolysaccharide, followed by incubation for 3 hours, the compound of example 1 and lipopolysaccharide were both in solution in DMEM medium. The medium was removed, cells were collected, total RNA was extracted from RAW264.7 using RNAiso Plus, then RNA was reverse transcribed into cDNA using a reverse transcription kit, and the effect of the test compound on inflammatory factor mRNA expression levels was detected using a real-time fluorescent quantitative PCR method. Through 2 -ΔΔCt The expression level of the relative RNA was calculated and normalized to β -action, and the gene specific primers used for qPCR are shown in Table (3). The experiments were run simultaneously with a blank (DMEM medium alone, no cells), a control (DMEM medium with cells), a model (LPS 1 μg/mL added) and a positive (dexamethasone 20 μM added).
TABLE 3 Gene specific primer sequences for qPCR
Conclusion of experiment: compared with the control group, the mRNA levels of inflammatory cytokines IL-1 beta, IL-6, COX-2 and iNOS in the LPS group are obviously up-regulated ### p<0.001). The compounds of the present invention significantly down-regulated the elevated mRNA levels of IL-1 beta, IL-6, COX-2 and iNOS in the model group compared to the model group, and thus the compounds of the present invention were effective in inhibiting the elevated gene expression levels of inflammatory mediators in RAW264.7 cells (fig. 14).
The embodiments described above and features of the embodiments herein may be combined with each other without conflict.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (6)
1. A tetracyclic triterpene compound has a structural formula:
2. the tetracyclic triterpene compound according to claim 1, wherein said compound has the structural formula:
3. a process for the preparation of a compound as claimed in claim 1 or 2, comprising the steps of:
s1, weighing and drying inonotus obliquus, crushing, leaching with ethanol, and concentrating the extracting solution under reduced pressure to obtain ethanol extract;
s2, dissolving the ethanol extract with methanol, then adding petroleum ether for extraction, and concentrating the methanol part under reduced pressure to obtain a methanol extract;
s3, dissolving the methanol extract again by using a mixed solution of water and methanol in a volume ratio of 9:1, then adding ethyl acetate for extraction, and concentrating an ethyl acetate part under reduced pressure to obtain an ethyl acetate extract;
s4, carrying out normal phase silica gel column chromatography on the ethyl acetate extract, and carrying out gradient elution to obtain A-N14 components;
s5, performing medium-pressure reverse phase column chromatography on the obtained K components, and performing gradient elution to finally obtain K-1-K-1414 components;
s6, separating the K-8 component by semi-preparation high performance liquid chromatography to obtain the compound;
the conditions of the gradient elution of the ethyl acetate extract are as follows: petroleum ether-ethyl acetate is used as an eluent, gradient elution is carried out according to the volume ratio of 20:1, 10:1, 9:1, 8:2, 7:3, 6:4, 1:1 and 0:1, and similar components are detected and combined by TLC, wherein the developing agent is a mixed solution of chloroform-toluene-ethyl acetate-methanol;
the conditions for the gradient elution of the component K are as follows: performing gradient elution by using water-methanol as an eluent according to the volume ratio of 90:10, 70:30, 50:50, 30:70 and 0:100, and detecting and combining similar components by TLC, wherein the developing agent is a mixed solution of chloroform-toluene-ethyl acetate-methanol;
the semi-preparative high performance liquid chromatography condition of the component K-8 is as follows: the mixed solution of acetonitrile and formic acid water solution is used as eluent, and the following steps are carried out: isocratic elution is carried out by the volume ratio of the formic acid aqueous solution to 40:60, the flow rate is 3mL/min, and the volume percentage of formic acid in the formic acid aqueous solution is 0.1%.
4. Use of a tetracyclic triterpene compound according to claim 1 or 2, in the manufacture of a medicament for inhibiting the production of NO.
5. Use of a tetracyclic triterpene compound according to claim 1 or 2, for the preparation of a medicament for inhibiting the expression level of an inflammatory factor mRNA.
6. Use of a tetracyclic triterpene compound according to claim 1 or 2 for the preparation of an anti-inflammatory medicament.
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Non-Patent Citations (2)
| Title |
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| Chemical Constituents from Inonotus obliquus and Their Biological Activities;Liu, Chao等;Journal of Natural Products;第77卷(第1期);35-41 * |
| 桦褐孔菌子实体三萜单体的分离及鉴定;陈程等;浙江理工大学学报;第70卷(第4期);721-730 * |
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