CN109293494A - The 1,4- naphthoquinone compound in a kind of mangrove endogenetic fungus source and preparation method thereof and application in preparing anti-inflammatory drugs - Google Patents
The 1,4- naphthoquinone compound in a kind of mangrove endogenetic fungus source and preparation method thereof and application in preparing anti-inflammatory drugs Download PDFInfo
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- CN109293494A CN109293494A CN201811080944.9A CN201811080944A CN109293494A CN 109293494 A CN109293494 A CN 109293494A CN 201811080944 A CN201811080944 A CN 201811080944A CN 109293494 A CN109293494 A CN 109293494A
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- naphthoquinone
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- 240000002044 Rhizophora apiculata Species 0.000 title claims abstract description 26
- -1 1,4- naphthoquinone compound Chemical class 0.000 title claims abstract description 23
- 241000233866 Fungi Species 0.000 title claims abstract description 21
- 229940124599 anti-inflammatory drug Drugs 0.000 title claims abstract description 13
- 238000002360 preparation method Methods 0.000 title claims description 15
- FRASJONUBLZVQX-UHFFFAOYSA-N 1,4-dioxonaphthalene Natural products C1=CC=C2C(=O)C=CC(=O)C2=C1 FRASJONUBLZVQX-UHFFFAOYSA-N 0.000 title claims description 8
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 25
- 238000000855 fermentation Methods 0.000 claims description 13
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- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
- 238000004587 chromatography analysis Methods 0.000 claims description 5
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- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 abstract description 7
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- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
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- CGIGDMFJXJATDK-UHFFFAOYSA-N indomethacin Chemical compound CC1=C(CC(O)=O)C2=CC(OC)=CC=C2N1C(=O)C1=CC=C(Cl)C=C1 CGIGDMFJXJATDK-UHFFFAOYSA-N 0.000 description 4
- 239000013641 positive control Substances 0.000 description 4
- QAPSNMNOIOSXSQ-YNEHKIRRSA-N 1-[(2r,4s,5r)-4-[tert-butyl(dimethyl)silyl]oxy-5-(hydroxymethyl)oxolan-2-yl]-5-methylpyrimidine-2,4-dione Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O[Si](C)(C)C(C)(C)C)C1 QAPSNMNOIOSXSQ-YNEHKIRRSA-N 0.000 description 3
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- HTSGKJQDMSTCGS-UHFFFAOYSA-N 1,4-bis(4-chlorophenyl)-2-(4-methylphenyl)sulfonylbutane-1,4-dione Chemical compound C1=CC(C)=CC=C1S(=O)(=O)C(C(=O)C=1C=CC(Cl)=CC=1)CC(=O)C1=CC=C(Cl)C=C1 HTSGKJQDMSTCGS-UHFFFAOYSA-N 0.000 description 1
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
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- 241000862501 Kandelia candel Species 0.000 description 1
- 241000987443 Kandelia obovata Species 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C50/00—Quinones
- C07C50/26—Quinones containing groups having oxygen atoms singly bound to carbon atoms
- C07C50/32—Quinones containing groups having oxygen atoms singly bound to carbon atoms the quinoid structure being part of a condensed ring system having two rings
-
- 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]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/66—Preparation of oxygen-containing organic compounds containing the quinoid structure
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2602/00—Systems containing two condensed rings
- C07C2602/02—Systems containing two condensed rings the rings having only two atoms in common
- C07C2602/04—One of the condensed rings being a six-membered aromatic ring
- C07C2602/10—One of the condensed rings being a six-membered aromatic ring the other ring being six-membered, e.g. tetraline
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicines Containing Plant Substances (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of 1,4-naphthoquinone class compounds in mangrove endogenetic fungus source, and the structural formula of the 1,4-naphthoquinone class compound is as shown in Formulas I and II.The NO that such compound generates LPS induction 264.7 mouse macrophage of RAW has certain inhibiting effect, IC50Value is respectively 3.9µM and 1.7µM.Further study show that compound ii can inhibit the mRNA expression of the relevant inductivity synthase of inflammation such as iNOS, COX-2, TNF-α, IL-6 the and IL-1 β of LPS induction, the expression of the iNOS and COX-2 albumen of LPS induction may also suppress.Illustrate that compound I and II has the function of inhibiting Macrophage Inflamatory reaction, there is good extracorporeal anti-inflammatory activity, can be used for preparing anti-inflammatory drug.Therefore, 1,4-naphthoquinone class compound provided by the invention has anti-inflammatory clinical application potentiality.
Description
Technical field
The invention belongs to medical compounds technical fields, more particularly, to a kind of Isosorbide-5-Nitrae-in mangrove endogenetic fungus source
Naphthoquinone compound and preparation method thereof and application in preparing anti-inflammatory drugs.
Background technique
Inflammation is body tissue to a kind of defensive anti-of destructive stimulus (such as destructive stimulus, germ or physical damnification)
It answers.The pathological change of inflammation is mainly made of the rotten of local organization, exudation and hyperplasia this three parts, and symptom clinically is then
For red, swollen, hot, pain and local dysfunction.Inflammation is also the important factor in human aging process, such as with many chronic diseases
The close phases such as arthritis, osteoporosis, asthma, A Ercimo disease, cardiovascular disease, dementia, cancer, obesity and type II diabetes
It closes [1-3].Clinically, anti-inflammatory drug is the second major class drug for being only second to anti-infectives.Inflammation is the common of many diseases
Pathologic process, if body cannot remove inflammation-causing substance in time or acute inflammation is changed into chronic inflammation, inflammation protracted course of disease meeting
Seriously affect the function of body.Current clinically common anti-inflammatory agent mainly includes non-steroidal anti-inflammatory drugs and steroidal anti-inflammatory drugs.
Although these two types of anti-inflammatory agents have certain clinical antiphlogistic effects, long-term a large amount of uses can generate a series of adverse reactions and resistance to
By property, such as gastric mucosa damage, hepar damnification, kidney damage.For the tolerance and adverse reaction for solving drug, new resist is found
Scorching drug and the drug of related novel framework types become the hot spot of anti-inflammatory drug research field.
Macrophage is a kind of immunocyte important in body, is the main cell that internal starting inflammatory mediator generates,
And there is anti-infective, antitumor and immunoregulatory effect.Lipopolysaccharides (LPS) is the Gram-negative bacterial cells such as Escherichia coli
One of component of wall, it can stimulate body to cause violent inflammatory reaction with the various associated receptors on active cell film.It utilizes
LPS inducing macrophage generates the cell model that inflammatory reaction is current common research inflammation.
Natural products is the important sources of lead compound.The natural products in marine fungi source has sustainable
Property, environmentally friendly, the features such as metabolite is rich and varied, be always the important source of drug screening.The metabolite of fungi
Antitumor with extensive physiological activity, such as antibacterial, immunological regulation is anti-inflammatory, and enzyme inhibits etc..Currently, being found from marine fungi
New medicine source molecule has become the hot spot of international and domestic research.
Summary of the invention
The purpose of the present invention is to provide the 1,4- naphthoquinone compounds in two mangrove endogenetic fungus sources.
It is another object of the present invention to provide a kind of 1,4- naphthoquinone compounds in above-mentioned mangrove endogenetic fungus source
Preparation method.
It is yet a further object of the present invention to provide the 1,4- naphthoquinone compounds in above-mentioned mangrove endogenetic fungus source to make
Application in standby anti-inflammatory drug.
Above-mentioned technical purpose of the invention is achieved through the following technical solutions:
A kind of 1,4-naphthoquinone class compound in mangrove endogenetic fungus source, the structural formula of the 1,4-naphthoquinone class compound is such as
Formulas I (new) and shown in II:
It is described present invention simultaneously provides the preparation method of the 1,4-naphthoquinone class compound in the mangrove endogenetic fungus source
1,4- naphthoquinone compound is to separate to obtain from the fermentation liquid of mangrove endogenetic fungus Talaromyces sp.SK-S009；Institute
It states Talaromyces sp.SK-S009 bacterial strain and is preserved in Guangdong Province's Culture Collection, preservation on May 9th, 2018
Number is GDMCC No:60369.
Further, the method specifically comprises the following steps:
S1. mangrove endogenetic fungus Talaromyces sp.SK-S009 is accessed into seed culture medium, obtains seed culture fluid；
S2. seed culture fluid is accessed in fermentation medium and is cultivated, obtain tunning；
S3. thallus is obtained by filtration in tunning, thallus purifying is concentrated, and extraction affords institute using chromatography
1,4 naphthoquinone compounds in the mangrove endogenetic fungus source stated.
Further, the composition of seed culture fluid described in S1 is by weight are as follows: glucose 0.3%, yeast extract
0.1%, peptone 0.1%~0.5%, agar 1.5%~2.5%, sodium chloride 1.5%~4%, water 93~98%.
Further, the condition cultivated in S2 is to cultivate 4~8 days at 150~200rpm of revolving speed, 28~35 DEG C.
Further, the fermentation medium is solid rice fermentation culture medium, and solid rice fermentation culture medium is rice
It is mixed with water according to the mass ratio of 1:1.
Further, it is stood after being cultivated in S2, the condition of the standing is 25~35 DEG C and stands 1 month.
Further, medicinal extract is obtained after concentration, ethyl acetate extract is extracted with ethyl acetate to obtain in medicinal extract, by ethyl acetate
Crude extract is chromatographed with silica gel normal-phase chromatography and is separated；It is eluted with petrol ether/ethyl acetate, collects 10%~60% acetic acid
Ethyl ester/petroleum ether moiety, then column chromatography for separation is used to get 1,4-naphthoquinone class compound shown in Formulas I and II is arrived.
The present invention protects the 1,4- naphthoquinone compound in the mangrove endogenetic fungus source preparing anti-inflammatory drug simultaneously
In application.
It further, is to inhibit the application in anti-inflammatory drug in preparation.
The present invention compared with the existing technology, have following advantages and effects
The present invention provides the 1,4-naphthoquinone class compound in two mangrove endogenetic fungus sources, small to LPS induction RAW 264.7
The NO that mouse macrophage generates has certain inhibiting effect, IC50Value is respectively 3.9 and 1.7 μM.Further study show that chemical combination
Object II can inhibit the mRNA table of the relevant inductivity synthase of inflammation such as iNOS, COX-2, TNF-α, IL-6 the and IL-1 β of LPS induction
It reaches, may also suppress the expression of the iNOS and COX-2 albumen of LPS induction.Illustrate that compound I and II has and inhibits Macrophage Inflamatory
The effect of reaction has good extracorporeal anti-inflammatory activity, can be used for preparing anti-inflammatory drug, have anti-inflammatory clinical application potentiality.
Detailed description of the invention
Fig. 1 is compound ii to iNOS, TNF-α, COX-2, the influence of IL-1 β and IL-6mRNA expression quantity.
Fig. 2 is influence of the compound ii to iNOS and COX-2 expressing quantity.
Specific embodiment
Further illustrate the present invention below in conjunction with specific embodiments and the drawings, but embodiment the present invention is not done it is any
The restriction of form.Unless stated otherwise, the present invention uses reagent, method and apparatus is the art conventional reagents, method
And equipment.
Unless stated otherwise, agents useful for same and material of the present invention are commercially available.
Embodiment 1
The preparation method of the 1,4-naphthoquinone class compound in two mangrove endogenetic fungus sources, the 1,4-naphthoquinone class compound
It is isolated from the fermentation liquid of ankle section category fungi Talaromyces sp.SK-S009 raw in mangrove.Mangrove endogenetic fungus
Talaromyces sp.SK-S009 is separated from the fruit of Guangxi mountain pass Kandelia candel mangrove (Kandelia obovata)
It arrives.
The mangrove fungi Talaromyces sp.SK-S009 is deposited in Guangdong Province's microbial bacteria on May 9th, 2018
Kind collection (GDMCC), deposit number are GDMCC No:60369, and classification naming is Talaromyces sp..Depositary institution
Address be the compound the 59th of Xianlie Middle Road, Guangzhou City 100 5 building, building.
The 1,4- naphthoquinone compound specific the preparation method is as follows:
S1. the seed liquor culture of mangrove endogenetic fungus Talaromyces sp.SK-S009: by mangrove endogenetic fungus
Talaromycessp.SK-S009 accesses seed culture medium, and at shaking speed 180rpm, 30 DEG C are cultivated 6 days, obtains seed training
Nutrient solution；Seed culture medium forms by weight are as follows: glucose 0.3%, yeast extract 0.1%, peptone 0.5%, agar
2.5%, sodium chloride 3%, water 98%.
S2. the fermented and cultured of mangrove endogenetic fungus Talaromyces sp.SK-S009: the culture of solid rice fermentation is utilized
Base (rice: tap water=1:1) transfers the bacterial strain in seed culture fluid in fermentation medium, stands 1 in 30 DEG C of room temperature
Month；
S3. above-mentioned cultured thallus methanol is extracted 3 times, concentrated extracting solution, by the concentrated extract acetic acid of acquisition
Ethyl ester extraction, obtains ethyl acetate extract.Ethyl acetate extract is chromatographed with silica gel normal-phase chromatography and is separated；Use petroleum
Ether/ethyl acetate is eluted, and collects 10%~60% ethyl acetate/petroleum ether part, then with silica gel, gel, C-18 reverse phase
Etc. column chromatography for separation technology to get arrive chemical compounds I and II.
Embodiment 2 (compound structure characterization)
Structured testing parsing is carried out to chemical compounds I (new) and II, obtains following experimental data:
Molecular formula C15H16O5, HRESI-MS:275.09223 [M-H]-(calculated value 275.09195)；
The NMR data of this compound is shown in Table 1.
1 chemical compounds I of table and II NMR data (500/125MHz, CDCl3)
According to above-mentioned data result, confirm chemical compounds I and II structural formula it is as follows:
Embodiment 3
The anti-inflammatory cells screening model of compound
1, the culture and processing of cell
264.7 cell of in vitro culture RAW, using the DMEM high glucose medium containing 10%FBS, in 37 DEG C, 5% titanium dioxide
Conventional maintenance culture and passage are carried out under the conditions of concentration of carbon.
2, compound intervention
Adjusting 264.7 cell density of RAW is 1 × 105The hole cells/ is simultaneously in logarithmic growth phase, and LPS (final concentration 1 is added
μ g/mL) inducing macrophage is in inflammatory conditions, and it is dense that untested compound or Indomethacin be made into different drugs using DMSO
Degree, each concentration set 3 parallel multiple holes, and Positive control wells (only plus LPS) is arranged, negative control hole (cell and culture medium),
Blank control wells (culture medium).After culture 24 hours, 50 μ L of cell supernatant is taken to be added in 96 new orifice plates, NO detection examination is added
Each 50 μ L of reagent I and II of agent box utilizes the content of Griess method measurement NO.
Inhibiting rate=([NO2-]LPS-[NO2-]LPS+sample)/([NO2-]LPS-[NO2-]untreated) × 100%
3, the influence of compound on intracellular vigor
MTT can be reduced to the blue crystallization of water-insoluble by the succinate dehydrogenase in living cells mitochondria, and be deposited on
In living cells, and dead cell has no this function.
264.7 cell of RAW of logarithmic growth phase, by 1 × 105The hole cells/ is inoculated in 96 orifice plates, the 100 every holes μ L.Add
Enter LPS (1 μ g/mL of final concentration) inducing macrophage and be in inflammatory conditions, is matched untested compound or Indomethacin using DMSO
At different drug concentrations, each concentration sets 3 parallel multiple holes, and Positive control wells (only plus LPS), negative control hole is arranged
(cell and culture medium), blank control wells (culture medium).After culture 24 hours, the 50 μ L of MTT solution of 1mg/mL, training is added in every hole
After supporting 4 hours, culture medium is sucked out, in the DMSO that 150 μ L are added to every hole.Concussion shakes up.It is measured with microplate reader in 490nm wavelength
The absorbance value (A) at place.The inhibiting effect that drug grows cell indicates that survival rate is higher with survival rate, indicates drug toxicity
It is lower.
Survival rate (%)=[(A1–A0)/(A2-A0)] × 100%, wherein A0For the absorbance value of blank control, A1For sample
The absorbance value of product, A2For the absorbance value of Positive control wells.The sample of 5 concentration is measured, dosage-inhibiting rate curve is drawn,
Obtain its CC50Value.Each sample is repeated three times.The selection result such as table 2.
2 chemical compounds I of table and II NO inhibitory activity and cytotoxicity assay result
aPositive control
4, compound ii is to iNOS, COX-2, the expression of IL-1 β, IL-6 and TNF-α in mRNA level in-site
The RAW264.7 cell inoculation of logarithmic growth phase is randomly divided into Normal group, model in six porocyte plates
Group and administration group, every group sets 3 multiple holes.LPS (1 μ g/ of final concentration is added in Normal group complete culture solution culture, model group
ML it) is stimulated, administration group is separately added into LPS (1 μ g/mL of final concentration) and drug, incubator culture 12h discard culture solution, add
Enter 1ml trizol cracking, extract total serum IgE, carry out reverse transcription reaction, further utilize TNF-α and NO primer, carries out PCR expansion
Increase, PCR product carries out separation identification with 2% agaropectin, and wherein GAPDH measures compound as internal reference, real-time fluorescence quantitative PCR
II couple of iNOS, COX-2, the inhibiting effect of IL-1 β, IL-6 and TNF-α mRNA gene expression.
3 fluorescent quantitation primer gene order table of table
The normal cell mrna expression amount that unused LPS is induced is set as 1, the IL-1 β expression quantity after LPS is induced significantly mentions
Height, the compound ii of various concentration can inhibit the iNOS induced by LPS, TNF-α, COX-2, the mRNA water of IL-1 β and IL-6
Flat (P < 0.05), and the inhibitory effect of high concentration is better than low concentration, the result is shown in Figure 1.
5, Western Western blot detection compound II to iNOS, COX-2 protein level expression
The RAW264.7 cell inoculation of logarithmic growth phase is randomly divided into Normal group, model in six porocyte plates
Group and administration group, every group sets 3 multiple holes.LPS (1 μ g/ of final concentration is added in Normal group complete culture solution culture, model group
ML it) being stimulated, administration group is separately added into LPS (1 μ g/mL of final concentration) and drug, and incubator culture 12h, PBS wash cell,
Cell is collected, cell pyrolysis liquid is added, half an hour is incubated under the conditions of 4 DEG C, 12,000rpm centrifugations 15 minutes take supernatant, take 10 micro-
It rises and carries out protein quantification, 2 × loadingbuffer of remaining addition equivalent, boiling water boiling 5 minutes, 10%SDS-PAGE (100v)
Separation, in transferring film to nitrocellulose filter, 5%BSA (TBS-T buffer) room temperature is closed 1 hour, i NOS and COX-2 antibody in
It is incubated overnight under the conditions of 4 DEG C, TBS-T is washed three times, is incubated at room temperature 1.5h with corresponding HRP- secondary antibody, and TBS-T is washed three times, and hair is added
Light reagent, colour developing imaging.
Western immune-blotting method the result shows that, not plus in the case where LPS, iNOS, COX-2 albumen yield
It is extremely low, and the yield both being added after LPS dramatically increases.After the compound ii processing of various concentration, iNOS and COX-2 egg
White expression quantity significantly reduces, and high dose is better than low dosage (see Fig. 2).
Claims (10)
1. a kind of 1,4-naphthoquinone class compound in mangrove endogenetic fungus source, which is characterized in that the 1,4-naphthoquinone class compound
Structural formula as shown in Formulas I and II:
。
2. a kind of preparation method of the 1,4-naphthoquinone class compound in mangrove endogenetic fungus described in claim 1 source, feature
It is, the 1,4-naphthoquinone class compound is from mangrove endogenetic fungusTalaromycesSp. it is separated in the fermentation liquid of SK-S009
It obtains；It is describedTalaromycesSp. SK-S009 bacterial strain is preserved in Guangdong Province's Microbiological Culture Collection on May 9th, 2018
Center, deposit number are GDMCC No:60369.
3. preparation method according to claim 2, which is characterized in that specifically comprise the following steps:
S1. by mangrove endogenetic fungusTalaromycesSp. SK-S009 accesses seed culture medium, obtains seed culture fluid；
S2. seed culture fluid is accessed in fermentation medium and is cultivated, obtain tunning；
S3. thallus is obtained by filtration in tunning, thallus purifying is concentrated, extraction, affords using chromatography described
Mangrove endogenetic fungus source 1,4- naphthoquinone compound.
4. the preparation method according to claim 3, which is characterized in that the composition of seed culture fluid described in S1 is by weight
Than are as follows: glucose 0.3%, yeast extract 0.1%, peptone 0.1%~0.5%, agar 1.5%~2.5%, sodium chloride 1.5%~
4%, water 93~98%.
5. preparation method according to claim 3, which is characterized in that the condition cultivated in S2 is in revolving speed 150~200
Rpm is cultivated 4~8 days at 28~35 DEG C.
6. preparation method according to claim 3, which is characterized in that the fermentation medium is the culture of solid rice fermentation
Base, solid rice fermentation culture medium are that rice is mixed with water according to the mass ratio of 1:1.
7. preparation method according to claim 3, which is characterized in that stood after being cultivated in S2, the condition of the standing is
25~35 DEG C stand 1 month.
8. the preparation method according to claim 3, which is characterized in that obtain medicinal extract, medicinal extract ethyl acetate after concentration
Ethyl acetate extract is extracted to obtain, ethyl acetate extract is chromatographed with silica gel normal-phase chromatography and is separated；With petroleum ether/acetic acid
Ethyl ester is eluted, and 10%~60% ethyl acetate/petroleum ether part is collected, then uses column chromatography for separation to get Formulas I institute is arrived
The 1,4- naphthoquinone compound shown.
It is answered 9. the 1,4- naphthoquinone compound in mangrove endogenetic fungus described in claim 1 source is in preparing anti-inflammatory drugs
With.
10. application according to claim 9, which is characterized in that be application in preparing anti-inflammatory drugs.
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