CN104402899B - Citrinin compounds penicitrinol L coming from Aspergillus citrimum and its preparation method and application - Google Patents
Citrinin compounds penicitrinol L coming from Aspergillus citrimum and its preparation method and application Download PDFInfo
- Publication number
- CN104402899B CN104402899B CN201410782653.XA CN201410782653A CN104402899B CN 104402899 B CN104402899 B CN 104402899B CN 201410782653 A CN201410782653 A CN 201410782653A CN 104402899 B CN104402899 B CN 104402899B
- Authority
- CN
- China
- Prior art keywords
- compound
- dichloromethane
- preparation
- ibpt
- aspergillus citrimum
- 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.)
- Expired - Fee Related
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/06—Peri-condensed systems
-
- 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
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
- C12P17/181—Heterocyclic compounds containing oxygen atoms as the only ring heteroatoms in the condensed system, e.g. Salinomycin, Septamycin
Abstract
The present invention relates to a kind of citrinin compounds penicitrinol L coming from Aspergillus citrimum and its preparation method and application.This compound has suppression tumor cell proliferation effect, has anti-tumor activity.Its structural formula is:.By fermentation culture Aspergillus citrimum (<i>penicillium citrinum</i>) IBPT-5, obtain fermented product, then separate from fermented product and be purified into this compound.Being verified by experiments, Human cervical cancer cell lines hela is had good anti-tumor activity by this compound.Can study for antineoplastic as preparing cytostatic thing or antitumor drug.
Description
Technical field
The present invention relates to a kind of citrinin compounds penicitrinolL coming from Aspergillus citrimum and its preparation method and application.
Background technology
Fungus is relatively high in microorganism.Fungus natural product is the important sources of activity multiformity and structure diversity, has been developed over many chemicalses with good therapeutical effect.Such as, antibiotics penicillin (penicillin) series, immunosuppressant cyclosporin A (cyclosporinA), hypolipidemic mevastatin (mevastatin), lovastatin (lovastatin) and antifungal drug griseofulvin (griseofulvin) etc., these all illustrate that fungus is the important treasure-house of natural drug.Citrinin, is a kind of famous polyketone class fungal secondary metabolite, is separate to obtain from the secondary metabolite of a strain penicillium citrinum the earliest.Finding in research in recent years, citrinin compounds shows the biologic activity such as gratifying antitumor, antibacterial, antioxidation, fouling resistance, enzyme level.
The present inventor's research is learnt, Aspergillus citrimum (Penicilliumcitrinum) IBPT-5, (within 25th, it is deposited in China typical culture collection center in December in 2013, address: Wuhan Wuhan University, deposit number is: CCTCCNO:M2013713) the crude extract of tunning have good cell inhibitory effect activity, then its active component is studied.Research finds that shown alkaloid compound has anti-tumor activity, has not yet to see the report of the proliferation inhibition activity of this compound on tumor cell, therefore also there is not yet medicine related to this on market.
Summary of the invention
It is an object of the invention to provide a kind of citrinin compounds penicitrinolL coming from Aspergillus citrimum and its preparation method and application.This compound has suppression tumor cell proliferation effect, has anti-tumor activity.Its structural formula is:
。
The preparation method of this compound, it is by fermentation culture Aspergillus citrimum (Penicilliumcitrinum) IBPT-5, obtain fermented product, then separate from fermented product and be purified into this compound, described Aspergillus citrimum (Penicilliumcitrinum) IBPT-5, within 25th, being deposited in China typical culture collection center in December in 2013, address: Wuhan Wuhan University, deposit number is: CCTCCNO:M2013713.Specifically comprise the following steps that
1 fermenting and producing
The conventional method of cultivating microorganism, take Aspergillus citrimum (Penicilliumcitrinum) IBPT-5 to be inoculated on PDA solid slant culture base in 28 DEG C of incubators and cultivate 4 days, being then seeded in culture fluid, 28 DEG C of static gas wave refrigerator are after 30 days, it is thus achieved that mycelium and fermentation liquid；Described culture fluid forms: every liter of water is containing mannitol 20.0g, yeast extract 3.0g, maltose 20.0g, monosodium glutamate 10.0g, glucose 10.0g, KH2PO40.5g、MgSO40.3g, NaCl30.0g；
The acquisition of 2 extractum
With gauze by mycelium and separation of fermentative broth.By the fermentation liquid extraction into ethyl acetate twice of 2 times of volumes, extract decompression is distilled to dry, obtains the ethyl acetate extract of fermentation liquid.
The separation and purification of 3 compounds
This ethyl acetate extract is by after 100-200 order silica gel mixed sample, with petroleum ether: dichloromethane: methanol reduces pressure silica gel chromatographic column gradient elution for eluent, collect the eluate of dichloromethane, with dichloromethane: methanol is for eluent, further by pressurized silica gel column chromatography gradient elution, obtain dichloromethane: the eluate of methanol v/v=100:1, again through semi-preparative liquid chromatography 1010 type ODS-A, 10 × 250mm, 5 μm: separation flow velocity is 5mL/min, mobile phase is that 70% acetonitrile is containing 0.1%TFA, obtain described compound, tR16.7min。
The present invention also protects described compound purposes in preparing Cytostatic to tumor cell medicine and the purposes that this compound is in preparing antitumor drug.Described tumor cell is hela cell.
The remarkable advantage of the present invention: this citrinin compound shown in research is comparatively rare, Human cervical cancer cell lines hela is had significant anti-tumor activity by described citrinin compounds, can study for antineoplastic as preparing cytostatic thing or antitumor drug.Have not yet to see the report of this compound on tumor cell inhibitory effect activity, therefore market also there is not yet medicine related to this.
Accompanying drawing explanation
Fig. 1 is COSY, HMBC and NOE signal main for compound penicitrinolL.
Detailed description of the invention
The chemical constitution of the compound of indication in examples below:
。
The fermenting and producing of this compound of embodiment 1 and separation and purification
1 fermenting and producing
Produce the fermentation culture of bacterium: by the conventional method of cultivating microorganism, take Aspergillus citrimum (Penicilliumcitrinum) IBPT-5 and (within 25th, be deposited in China typical culture collection center in December in 2013, address: Wuhan Wuhan University, deposit number is: CCTCCNO:M2013713) appropriate, it is inoculated on PDA solid slant culture base in 28 DEG C of incubators and cultivates 4 days.
Take the slant culture Aspergillus citrimum of 4 days (Penicilliumcitrinum) IBPT-5 appropriate, be inoculated into equipped with 400mL culture fluid [culture fluid composition (g/l): mannitol 20.0, yeast extract 3.0, maltose 20.0, monosodium glutamate 10.0, glucose 10.0, KH2PO40.5, MgSO40.3, NaCl30.0 constant volume] 1000mL conical flask in, 28 DEG C of static gas wave refrigerator are after 30 days, it is thus achieved that mycelium and fermentation liquid.
The acquisition of 2 extractum
With gauze by mycelium and separation of fermentative broth.By fermentation liquid ethyl acetate 1:2 (v/v) extracting twice, extract decompression is distilled to dry, obtains the ethyl acetate extract 32g of fermentation liquid.
The separation and purification of 3 compounds
This extractum is by after 100-200 order silica gel mixed sample, with petroleum ether: dichloromethane: methanol, for eluent decompression silica gel chromatographic column gradient elution, obtains 11 components.Component 3 (1.8g) (eluate of dichloromethane) is with dichloromethane: methanol is for eluent, further by pressurized silica gel column chromatography gradient elution, the subfraction 3-1 (350mg) (dichloromethane: the eluate of methanol v/v=100:1) obtained, again through semi-preparative liquid chromatography (1010 type ODS-A, 10 × 250mm, 5 μm): separation flow velocity is 5mL/min, mobile phase is that 70% acetonitrile is containing 0.1%TFA, obtain shown compound (7.2mg, tR16.7min)。
Compound as white powder, high resolution mass spectrum HRESI-MS provides molecular ion peak [M H] at m/z361.1671 place–, (calcd.forC20H25O6, 361.1657), prompting molecular weight is 362, speculates that molecular formula is C in conjunction with spectral information20H26O6。1H and13C-NMR data are in Table 1, and main COSY, HMBC and NOE signal is shown in Fig. 1.
Table 1 compound1H and13C-NMR data (500MHz1Hand125MHz13C, inCDCl3)
The test of embodiment 2 anti tumor activity in vitro
1 laboratory sample and experimental technique
The preparation test sample of sample solution is the pure compounds of separation and purification in above-mentioned enforcement 1.Precision weighs appropriate amount of sample, is configured to the solution of desired concn with methanol, for surveying activity.
The successive transfer culture of cell line and cell adopts tumor cell line, and the tumor cell DMEM culture medium containing 10%FBS, at 37 DEG C of successive transfer culture in the incubator passing into 5% carbon dioxide.
Cell inhibitory effect activity test method
Tetrazolium (MTT) method is taken the logarithm the tumor cell of trophophase, and cell density is adjusted to every milliliter 2 × 105Individual cell, is inoculated in 96 porocyte culture plates by every hole 200 microlitre, passes into 5%CO in 37 DEG C2Incubator in cultivate 4 hours.Every hole adds 2 microliters of sample liquid or blank solutions, and after cultivating 24 hours, every hole adds MTT liquid (5 milligrams of normal saline solutions of every milliliter of MTT) 10 microlitres, continues to cultivate 4 hours, and 37 DEG C, 2000 revs/min are centrifuged 8 minutes, suck supernatant.Every hole adds each 100 microlitres of DMSO, vibrates 15 minutes on micro oscillator, after being completely dissolved to crystallization, utilizes MD company to produce SPECTRAMAXPlus type microplate reader and measures every hole light absorption value (OD) value at 570nm place.In same 96 orifice plate, each concentration of sample is respectively provided with three holes, separately sets three hole blanks and acellular zeroing hole (if medicine has color to do the acellular zeroing of relative medicine concentration).Each hole OD value first does corresponding acellular zeroing, then takes three hole mean OD value by IR (%)=(ODBlank-ODSample)/ODBlank× 100% formula calculates cell proliferation inhibition rate (IR%) under each concentration.
2. experimental result
Cell inhibitory effect active testing result
In mtt assay is tested, the Cytostatic to tumor cell rate according to this compound of variable concentrations, application SPSS16.0 software carries out data and processes and calculation of half inhibitory concentration IC50Value.Result is in Table 2.
The inhibitory activity that human tumor cells is bred by table 2 compound
3. conclusion
Compound on tumor cell hela has stronger inhibited proliferation, can study for antineoplastic as preparation hela inhibition of cell proliferation or antitumor agent.
Claims (5)
1. compound。
2. the preparation method of compound as claimed in claim 1, it is characterized in that: fermentation culture Aspergillus citrimum (Penicilliumcitrinum) IBPT-5, obtain fermented product, then separate from fermented product and be purified into this compound, described Aspergillus citrimum (Penicilliumcitrinum) IBPT-5, within 25th, being deposited in China typical culture collection center in December in 2013, address: Wuhan Wuhan University, deposit number is: CCTCCNO:M2013713；
Specifically comprising the following steps that of described preparation method
(1) conventional method of fermenting and producing cultivating microorganism, take Aspergillus citrimum (Penicilliumcitrinum) IBPT-5 to be inoculated on PDA solid slant culture base in 28 DEG C of incubators and cultivate 4 days, it is then seeded in culture fluid, 28 DEG C of static gas wave refrigerator are after 30 days, it is thus achieved that mycelium and fermentation liquid；Described culture fluid forms: every liter of water is containing mannitol 20.0g, yeast extract 3.0g, maltose 20.0g, monosodium glutamate 10.0g, glucose 10.0g, KH2PO40.5g、MgSO40.3g, NaCl30.0g；
(2) the acquisition gauze of extractum is by mycelium and separation of fermentative broth, and by the fermentation liquid extraction into ethyl acetate twice of 2 times of volumes, extract decompression is distilled to dry, obtains the ethyl acetate extract of fermentation liquid；
(3) this ethyl acetate extract of the separation and purification of compound is by after 100-200 order silica gel mixed sample, with petroleum ether: dichloromethane: methanol reduces pressure silica gel chromatographic column gradient elution for eluent, collect the eluate of dichloromethane, with dichloromethane: methanol is for eluent, further by pressurized silica gel column chromatography gradient elution, obtain dichloromethane: the eluate of methanol v/v=100:1, again through semi-preparative liquid chromatography 1010 type ODS-A, 10 × 250mm, 5 μm: separation flow velocity is 5mL/min, mobile phase is that 70% acetonitrile is containing 0.1%TFA, obtain described compound, tR16.7min.
3. the purposes in preparing Cytostatic to tumor cell medicine of the compound described in claim 1.
4. the purposes in preparing antitumor drug of the compound described in claim 1.
5. purposes according to claim 3, it is characterised in that: described tumor cell is hela cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410782653.XA CN104402899B (en) | 2014-12-18 | 2014-12-18 | Citrinin compounds penicitrinol L coming from Aspergillus citrimum and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410782653.XA CN104402899B (en) | 2014-12-18 | 2014-12-18 | Citrinin compounds penicitrinol L coming from Aspergillus citrimum and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104402899A CN104402899A (en) | 2015-03-11 |
| CN104402899B true CN104402899B (en) | 2016-06-29 |
Family
ID=52640578
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410782653.XA Expired - Fee Related CN104402899B (en) | 2014-12-18 | 2014-12-18 | Citrinin compounds penicitrinol L coming from Aspergillus citrimum and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104402899B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107325081B (en) * | 2016-05-27 | 2019-07-09 | 福州大学 | Citrinin compounds dicitrinone D preparation method and its application in terms of lung cancer |
| CN107325086B (en) * | 2016-05-27 | 2019-07-09 | 福州大学 | Citrinin compounds dicitrinone D preparation method and its application in terms of colon cancer |
| CN107325088B (en) * | 2016-05-27 | 2019-07-09 | 福州大学 | Citrinin compounds dicitrinone D preparation method and the application in terms of nasopharyngeal carcinoma |
| CN107325085B (en) * | 2016-05-27 | 2019-07-09 | 福州大学 | Citrinin compounds dicitrinone D preparation method and its application in terms of lymph cancer |
| CN107325087B (en) * | 2016-05-27 | 2019-07-09 | 福州大学 | Citrinin compounds dicitrinone D and its application in terms of malignant mela noma |
| CN110790660B (en) * | 2019-09-26 | 2022-05-24 | 浙江工业大学 | Polyketide, preparation method, bacterial strain and application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101735236A (en) * | 2008-11-10 | 2010-06-16 | 中国海洋大学 | Dimer citrinin compounds and preparation and use thereof |
-
2014
- 2014-12-18 CN CN201410782653.XA patent/CN104402899B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101735236A (en) * | 2008-11-10 | 2010-06-16 | 中国海洋大学 | Dimer citrinin compounds and preparation and use thereof |
Non-Patent Citations (3)
| Title |
|---|
| Citrinin Dimers from the Halotolerant Fungus Penicillium citrinum B-57;Zhen-Yu Lu，等;《J. Nat. Prod.》;20080219;第71卷(第4期);第543-546页 * |
| Citrinin revisited:from monomers to dimers and beyond;Benjamin R.Clark，等;《Organic & Biomolecular Chemistry》;20060316;第4卷;第1520-1528页 * |
| New citrinin derivatives isolated from Penicillium citrinum;Daigo Wakana，等;《J.Nat.Med.》;20060405;第60卷(第4期);第279-284页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104402899A (en) | 2015-03-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104478891B (en) | Citrinin compounds penicitrinol O coming from Aspergillus citrimum and its preparation method and application | |
| CN103865808B (en) | A kind of anticancer usage of the penicillium sp enol A1 coming from Aspergillus citrimum | |
| CN104402898B (en) | Citrinin compounds penicitrinol M coming from Aspergillus citrimum and its preparation method and application | |
| CN104402899B (en) | Citrinin compounds penicitrinol L coming from Aspergillus citrimum and its preparation method and application | |
| CN104447781B (en) | Citrinin compounds penicitrinol N coming from Aspergillus citrimum and its preparation method and application | |
| CN104531540A (en) | Antitumor application of penicillium enol A2 from penicillium citrinum | |
| CN104592082B (en) | Come from the penicillium sp enol D of Aspergillus citrimum2preparation method and applications | |
| CN109106702A (en) | Derived from application of 4-4 ' the isomerization secalonic acid D in terms of colon cancer of penicillium oxalicum | |
| CN105061446B (en) | Penicillium citrinum-derived penicitrinine A as well as application thereof to preparation of drugs for resisting nasopharyngeal carcinoma | |
| CN103265522B (en) | Lactone derivative derived from trichoderma citrinoviride and application thereof | |
| CN104370928B (en) | Come from indole terpene speradine F and the application of aspergillus oryzae | |
| CN104447475B (en) | Preparation method and application of penicillenol D1 derived from penicillium citrinum | |
| CN105061443B (en) | Penicillium citrinum-derived penicitrinine A as well as application thereof to preparation of drugs for resisting human liver cancer | |
| CN106432035A (en) | Application of penem enol E1 derived from trichoderma citrinoviride on the aspect of malignant melanoma | |
| CN105001228B (en) | Application of penicillium citrinum-derived penicitrinine A in preparing drugs for treating human oral epidermoid tumor | |
| CN105061444B (en) | Penicillium citrinum-derived penicitrinine A as well as application thereof to preparation of drugs for resisting human colorectal carcinoma | |
| CN105153176B (en) | The penicitrinine A for coming from Penicillium citrinum and its application for preparing anti-human lung-cancer medicament | |
| CN105061445B (en) | Penicillium citrinum-derived penicitrinine A as well as application thereof to preparation of drugs for resisting human gastric cancer | |
| CN105153175B (en) | The penicitrinine A for coming from Penicillium citrinum and its application for preparing anti-human oesophagus cancer drug | |
| CN105131006B (en) | Penicitrinine A sourced from penicillium citrinum and application thereof in preparation of anti-malignant melanoma drug | |
| CN104211630B (en) | A kind of alkaloid compound and application thereof that comes from tangerine green trichoderma | |
| CN105017272B (en) | Penicitrinine A originated from penicillium citrinum and application thereof in preparation of anti human breast cancer drugs | |
| CN106491597A (en) | Come from applications of the mould enol E1 of tangerine green trichoderma in terms of the cancer of the esophagus | |
| CN106491596A (en) | Come from applications of the mould enol E1 of tangerine green trichoderma in terms of lung cancer | |
| CN107325086A (en) | Citrinin compounds dicitrinone D preparation methods and its application in terms of colon cancer |
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 |
Granted publication date: 20160629 Termination date: 20211218 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |