CN101481379A - Compound separated from acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain fermentation liquor - Google Patents
Compound separated from acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain fermentation liquor Download PDFInfo
- Publication number
- CN101481379A CN101481379A CNA2009100208206A CN200910020820A CN101481379A CN 101481379 A CN101481379 A CN 101481379A CN A2009100208206 A CNA2009100208206 A CN A2009100208206A CN 200910020820 A CN200910020820 A CN 200910020820A CN 101481379 A CN101481379 A CN 101481379A
- Authority
- CN
- China
- Prior art keywords
- compound
- fermented liquid
- ginko
- bacterial strain
- acetic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a compound separated from ethyl acetate extract of Chaetomium globosum fermentation liquor, and the structural formula of the compound is on the right. The compound is named as chaetomugilin D, the molecular formula is C23H27O6Cl, and the molecular weight is 435. Identification tests and cytotoxic activity tests by the applicant prove that the compound has very high cytotoxic activity on the brine shrimp, and can be used for preparing anticancer drugs.
Description
Technical field
The present invention relates to a kind of compound, the compound that is separated in particularly a kind of acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain Chaetomiumglobosum fermented liquid, this compound is a new compound, and shrimp ovum brine shrimp causes death to give birth to survey and shows very high cytotoxic activity.
Background technology
Cancer is the world-famous puzzle that can not capture always, and seeking anticancer active constituent from natural product is one of effective way that solves this world-famous puzzle.
Ginkgo is one of the most ancient higher plant, is the endemic plant of China, is used as traditional Chinese medicine and is developed to the modern health product because of the various activeconstituentss that it was rich in, especially flavonoid compound.According to " gene level transmission hypothesis " and " symbiosis theory ", may produce identical or similar chemical ingredients with ginkgo with the symbiotic endogenetic fungus of ginkgo.The pre-test of 98 strain gingko endogenous fungus chemical ingredientss display part endogenetic fungus as a result has the chemical ingredients identical with similar with ginkgo, as triterpene and flavonoid compound, system's prerun of endophyte Chaetomium globosum fermented product shows that also it can produce triterpene flavonoid and alkaloids substance, and most these materials all are physiologically active substances, for example antitumour activity.Therefore bacterial strain Chaetomium globosum has been carried out the secondary metabolism chemical constitution study, provide scientific basis for from gingko endogenous bacterium, seeking anti-cancer active matter.
Summary of the invention
The objective of the invention is to, isolating new compound in a kind of acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain Chaetomium globosum fermented liquid is provided, this compound has antitumour activity, can be used as the preparation cancer therapy drug and is employed.
In order to realize above-mentioned task, the present invention takes following technical solution:
Isolated compound in a kind of acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain Chaetomium globosum fermented liquid is characterised in that its structural formula is:
The name of this compound is called chaetomugilin D, and molecular formula is C
23H
27O
6Cl, molecular weight are 435.
Prove that through evaluation and biological activity test that the applicant carried out this compound prawn ovum brineshrimp has very high cytotoxic activity, can be used in the application of preparation cancer therapy drug.
Description of drawings
Fig. 1 is the acetic acid ethyl ester extract chemical ingredients extraction separation schema of Chaetomium globosum fermented liquid;
Fig. 2 be title compound chaetomugilin D crucial HMBC and
1H-
1H COSY characteristic relation;
Fig. 3-Figure 12 is the structure evaluation figure of title compound, wherein:
Fig. 3 is a title compound
1H NMR, Fig. 4 are title compounds
13C NMR, Fig. 5 and Fig. 6 are title compound HMBC; Fig. 7 is title compound H-H COSY; Fig. 8 is title compound HMQC, and Fig. 9 is title compound HSQC; Figure 10,11,12nd, title compound HMBC.
The present invention is described in further detail below in conjunction with accompanying drawing.
Embodiment
The present invention is for separating the new compound that obtains having antitumour activity in a kind of acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain Chaetomium globosum fermented liquid, host plant ginkgo (GinkgObiloba) derives from Shandong Province's Linyi City.The test that causes death of shrimp ovum is to finish in German brother's Dettingen university.Above-mentioned materials is disclosed biomaterial.
Isolation identification bacterial classification and the used culture medium prescription of expansion fermentation are respectively:
The PDA substratum is used for cultivating and separation purification of epiphyte, and its prescription is: potato juice 1000ml, glucose 20g, agar 20g, sterilization 30min.
Improved culture medium is used to enlarge liquid fermenting, and its prescription is: CaCl
2: 0.5g, KH
2PO
4: 0.1g, KCl:0.05g, MgSO
47H
2O:0.1g, sucrose: 20.0g, peptone: 15.0g, the H of 1000ml
2O, pH are 6.0.
The preparation method of potato juice is, potato decortication, and section claims 200g to put as going in the 1000ml tap water and boils 30min that double gauze filters, the filtrate moisturizing is to 1000ml.
The main reagent that utilizes is:
Sherwood oil (60-90 ℃), methylene dichloride, hexanaphthene, acetone, methyl alcohol etc. are analytical pure.
Isolated compound in the acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain Chaetomium globosum fermented liquid, its sepn process is as follows:
1. the fermentation of bacterial classification and fermented liquid concentrates
Draw the bacteria suspension 1ml of Chaetomium globosum, join 1000ml 200ml liquid nutrient medium (CaCl is housed
2: O.5g, KH
2PO
4: 0.1g, KCl:0.05g, MgSO
47H
2O:0.1g, glucose:20.0g, peptone:15.0g, H
2O:1000ml in triangular flask pH=6.0), at rotating speed 150rpm, cultivated 5 days down for 28 ℃, finished to ferment, and filtered and obtained fermented liquid 30L and mycelium, and mycelium is 50 ℃ of oven dry down, and average bacterium weighs 4.6g/L.
2. the preparation of crude extract
Filtrate is 30L altogether, uses equal volume of ethyl acetate 3 times, concentrates organic phase, obtains fermented liquid ethyl acetate extraction medicinal extract 2.83g;
3. the active prerun of crude extract
The sea shrimp causes death test because be the cytotoxicity screening method that the researchist trusted the most and widely using, (diameter is 1.8cm to concrete grammar for the shrimp ovum that 30 processes are selected is put into the culture tank that the 0.2ml synthetic sea water is housed, dark 2cm) in, dissolving 20 μ g fermented liquid ethyl acetate extracts with the DMSO of 5 μ l joins in the culture tank, contrast only adds the DMSO of equivalent, at dark surrounds, cultivate under the room temperature then.After 24 hours, add up shrimp ovum death toll in each culture tank, calculate lethality rate M with following formula at microscopically.
M=[(A-B-N)/(G-N)] 100, wherein A is the average death toll after 24 hours, and B is the average death toll of contrast the inside after 24 hours, and N is a shrimp ovum number dead before medicine adds, and G is total shrimp ovum number that is used to test.
4. the separation and purification of compound
Referring to Fig. 1, fermented liquid ethyl acetate extraction medicinal extract is by silica gel column chromatography, gel filtration chromatography, reversed phase column chromatography separation and Extraction chemical ingredients wherein, the normal pressure column chromatography adopts wet method dress post and silica gel mixed sample upper prop, select the eluting solvent system according to the thin-layer developing situation, reversed phase column chromatography adopts pressurizing device, gel filtration chromatography is selected Sephadex LH-20 for use, adopts methyl alcohol and methyl alcohol-methylene dichloride wash-out.Repeatedly column chromatography obtains compound at last.
Silicagel column: 2.83g fermented liquid ethyl acetate extraction medicinal extract dry method is mixed sample (100-200 order) 5.6g, 50g silica gel (200-300 order), with methylene chloride-methanol=100:0, methylene chloride-methanol=50:1, methylene chloride-methanol=30:1, methylene chloride-methanol=20:1, methylene chloride-methanol=10:1 gradient elution, every part of elutriant is 100ml, underpressure distillation concentrates, thin plate detects, similar merging; Obtain five components of A-E, wherein the B component shows stronger cytotoxic activity, so carry out double gel Sephadex LH-20 column chromatography under activity is followed the trail of, eluent is respectively (methylene chloride-methanol=6:4) and (methyl alcohol).Obtain active ingredient B2-3, continue silica gel column chromatography, obtain active ingredient B2-3-c, continue to use anti-phase RP-C then with hexanaphthene-acetone=10:1 wash-out
18Column chromatography purification obtains compound with water-methanol (30%, 45%, 50%, 55%) wash-out.
This compound is done thin-layer chromatography, and through ultraviolet, iodine and sulfuric acid-ethanol colour developing is single spot, and three kinds of solvent systemss launch to be single spot, can tentatively regard as the simplification compound.
5. the evaluation of compound
The compound that is separated to is done mass spectrum, carbon spectrum, hydrogen spectrum, and two dimensional NMR.Compound c haetomugilin D is an xanchromatic viscose glue shape, is dissolvable in water chloroform, methyl alcohol, and acetone, ethyl acetate, specific rotatory power are [α]
D 20-21 ° (c=0.04, MeOH).Drawing molecular formula by nuclear magnetic resonance spectrum data (seeing Table 1, Fig. 3-12) and high resolution mass spectrum is C
23H
27O
6Cl, molecular weight is 435, consults reference, the data that obtain of analysis-by-synthesis then, and, determine that according to physical property and SPECTROSCOPIC CHARACTERIZATION the title compound structure is at last in conjunction with the possible structure type of document check analysis compound:
Called after chaetomugilin D, structure is looked into newly through a plurality of databases, finds still not have the report of this compound both at home and abroad, can determine that this compound is new compound.
Table 1: NMR feature (500MHz, the CDCl of compound c haetomugilin D
3)
| No. | δC | δ H(J in Hz) | Selected HMBC | Selected |
| 1 | 145.6d | 7.27(1H，s) | C-8 | |
| 3 | 157.7s | C-4，1，9，10 | ||
| 4 | 104.9d | 6.55(1H，s) | C-9 | |
| 4a | 140.4s | C-1，8 | ||
| 5 | 110.1s | C-4， | ||
| 6 | 189.2s | C-8，7-Me | ||
| 7 | 84.0s | C-8，7-Me | ||
| 8 | 50.6d | 2.97(1H，d，10.1) | C-1，2’，7-Me | |
| 8a | 114.3s | C-8，1，4，2’ | ||
| 9 | 120.2d | 6.04(1H，d，15.6) | C-4，11 | |
| 10 | 146.9d | 6.52(1H，dd，15.6，6.5) | C-9，11，12 | |
| 11 | 38.9d | 2.23(1H，m，6.8) | C-9，10，12，13， 11-Me | |
| 12 | 29.2t | 1.42(2H，m) | C-11，10，13，11-Me | |
| 13 | 11.7q | 0.87(3H，t，7.3) | C-11，12 |
| 7-Me | 23.3q | 1.39(3H，s) | C-8 | |
| 11-Me | 19.4q | 1.10(3H，d，6.4) | C-10，11，12 | |
| 1’ | 170.5s | C-8，2’ | ||
| 2’ | 58.3d | 3.06(1H，d，10.1) | C-8 | |
| 3’ | 104.2s | C-4’，2’，5’，4’-Me | ||
| 4’ | 44.9d | 1.87(1H，dq，10.1，6.9) | C-6’，4’-Me | 2’-H，6’-H |
| 5’ | 76.9d | 4.28(1H，dq，10.1，6.4) | C-4’，6’，4’-Me | 8-H，7-Me |
| 6’ | 18.7q | 1.40(3H，d，6.4) | ||
| 4’-Me | 8.8q | 1.11(3H，d，6.2) | C-4’ |
6. the determination of cytotoxic activity of compound
By top crude extract antitumour activity measuring method, the test concentrations of pure compound is 10 μ g/ml, promptly dissolves 2 μ g fermented liquid ethyl acetate extracts with 5 μ l DMSO and joins in the culture tank, and the average dead lethality rate that the result obtained after 24 hours is 75.2%
In sum, the compound that is separated to from the acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain Chaetomium globosum fermented liquid has very strong antitumour activity.The sea shrimp causes death test-results for being under the 10 μ g/ml in concentration, and the average dead lethality rate after 24 hours is 75.2%.This compound is a kind of new compound, and its molecular formula is C
23H
27O
6Cl, molecular weight are 435.Called after chaetomugilin D.
Claims (4)
1. isolated compound in the acetic acid ethyl ester extract of a ginko endogenetic fungal bacterial strain Chaetomium globosum fermented liquid is characterized in that the structural formula of this compound is:
The name of this compound is called chaetomugilin D, and molecular formula is C
23H
27O
6Cl, molecular weight are 435.
2. the separation method of isolated compound in the acetic acid ethyl ester extract of the described ginko endogenetic fungal bacterial strain Chaetomium of claim 1 globosum fermented liquid is characterized in that, carries out as follows:
1) fermentation of bacterial classification and fermented liquid concentrates
Draw the bacteria suspension 1ml of ginko endogenetic fungal bacterial strain Chaetomium globosum, joining 1000ml is equipped with in the triangular flask of 200ml liquid nutrient medium, at rotating speed 150rpm, cultivated 5 days down for 28 ℃, finish fermentation, filtration obtains fermented liquid 30L and mycelium, and mycelium is 50 ℃ of oven dry down, and average bacterium weighs 4.6g/L;
2) preparation of crude extract
Filtrate is 30L altogether, uses equal volume of ethyl acetate 3 times, concentrates organic phase, obtains fermented liquid ethyl acetate extraction medicinal extract 2.83g;
3) separation and purification of compound
Fermented liquid ethyl acetate extraction medicinal extract is by silica gel column chromatography, gel filtration chromatography, reversed phase column chromatography separation and Extraction chemical ingredients wherein, the normal pressure column chromatography adopts wet method dress post and silica gel mixed sample upper prop, select the eluting solvent system according to the thin-layer developing situation, reversed phase column chromatography adopts pressurizing device, gel filtration chromatography is selected Sephadex LH-20 for use, adopt methyl alcohol and methyl alcohol-methylene dichloride wash-out, repeatedly column chromatography obtains compound c haetomugilin D at last.
3. method as claimed in claim 2 is characterized in that the prescription of described liquid nutrient medium is: CaCl
2: 0.5g, KH
2PO
4: 0.1g, KCl:0.05g, MgSO
47H
2O:0.1g, glucose:20.0g, peptone:15.0g, H
2O:1000ml, pH=6.0.
4. the application that isolated compound is used to prepare cancer therapy drug in the acetic acid ethyl ester extract of the described ginko endogenetic fungal bacterial strain Chaetomium of claim 1 globosum fermented liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100208206A CN101481379B (en) | 2009-01-07 | 2009-01-07 | Compound separated from acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain fermentation liquor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100208206A CN101481379B (en) | 2009-01-07 | 2009-01-07 | Compound separated from acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain fermentation liquor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101481379A true CN101481379A (en) | 2009-07-15 |
| CN101481379B CN101481379B (en) | 2011-03-23 |
Family
ID=40878689
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100208206A Expired - Fee Related CN101481379B (en) | 2009-01-07 | 2009-01-07 | Compound separated from acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain fermentation liquor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101481379B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103145725A (en) * | 2011-12-07 | 2013-06-12 | 上海来益生物药物研究开发中心有限责任公司 | Compound, and preparation method and application thereof |
| CN103694247A (en) * | 2013-12-06 | 2014-04-02 | 秦建春 | Compound Chaetomugilide A and preparation method and application thereof |
| CN109295122A (en) * | 2018-10-23 | 2019-02-01 | 华南农业大学 | A kind of Preparation method and use of E. exserta endogenetic fungus Chaetomium sp secondary metabolite |
| CN116115602A (en) * | 2022-12-23 | 2023-05-16 | 青岛农业大学 | New application of compound chaetomougilin O in preparation of medicaments for preventing or treating thyroid cancer |
-
2009
- 2009-01-07 CN CN2009100208206A patent/CN101481379B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103145725A (en) * | 2011-12-07 | 2013-06-12 | 上海来益生物药物研究开发中心有限责任公司 | Compound, and preparation method and application thereof |
| CN103145725B (en) * | 2011-12-07 | 2016-02-24 | 上海来益生物药物研究开发中心有限责任公司 | A kind of compound and its preparation method and application |
| CN103694247A (en) * | 2013-12-06 | 2014-04-02 | 秦建春 | Compound Chaetomugilide A and preparation method and application thereof |
| CN109295122A (en) * | 2018-10-23 | 2019-02-01 | 华南农业大学 | A kind of Preparation method and use of E. exserta endogenetic fungus Chaetomium sp secondary metabolite |
| CN116115602A (en) * | 2022-12-23 | 2023-05-16 | 青岛农业大学 | New application of compound chaetomougilin O in preparation of medicaments for preventing or treating thyroid cancer |
| CN116115602B (en) * | 2022-12-23 | 2024-03-05 | 青岛农业大学 | New application of compound chaetomougilin O in preparation of medicaments for preventing or treating thyroid cancer |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101481379B (en) | 2011-03-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112592350B (en) | Polyketide lithocarpin E-G and preparation method and application thereof | |
| CN109336873B (en) | Compound lithocarolsA-F, preparation method thereof and application thereof in preparation of antitumor drugs | |
| CN101481379B (en) | Compound separated from acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain fermentation liquor | |
| CN109232513A (en) | Compound lithocarpinols and preparation method thereof and application in preparation of anti-tumor drugs | |
| CN112358516B (en) | Application of diosmetin (4-O-methyl) glucoside compound in preparation of lipid-lowering drugs | |
| CN109942481B (en) | Compound Oleraisoindole A in purslane, and extraction separation method and application thereof | |
| CN105481817B (en) | A kind of isocoumarin class compound and its preparation method and application | |
| CN114213428B (en) | Indole alkaloid compound and preparation method and application thereof | |
| CN107501072B (en) | Compound colletotriconeA, preparation method thereof and application thereof in preparing antitumor drugs | |
| CN106279092B (en) | A kind of double 1,4-benzoquinone class compounds and its extracting method | |
| CN109265494A (en) | The method of Kaempferol glucoside compounds is extracted from the camellia of Yunnan | |
| CN105274152B (en) | Curcumin biotransformation method, product and application | |
| CN112794832B (en) | Compound NBY-10 extracted from folium Arctii and having antiinflammatory activity, and its preparation method and application | |
| CN104059038B (en) | A kind of sesquiterpenoids and application thereof | |
| CN106565444A (en) | Extraction method and application of phenanthrene compounds from overground part of Chinese yam | |
| CN101463062B (en) | Compound separated from Gynostemma pentaphylla total saponin acid hydrolysis products | |
| CN100522987C (en) | Saponins antineoplastic compound in Bohadschia argus Jaeger | |
| CN105837590A (en) | Compound with anti-Candida albicans activity, preparation method and application thereof | |
| CN111574362B (en) | Fatty acid compound | |
| CN114409557B (en) | Carbon keratin with neuroprotective activity and preparation method and application thereof | |
| CN114409625B (en) | Keratinone with neuroprotective activity and preparation method and application thereof | |
| CN115583953B (en) | Quinazolinone alkaloid compound, and preparation method and application thereof | |
| CN115894180B (en) | Method for separating mycotoxin from metabolite of black spot bacteria of ginseng | |
| CN101585809A (en) | Novel compound with antibacterial and antitumor activities | |
| CN110172065B (en) | Compound and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110323 Termination date: 20130107 |