CN110407798A - Derived from the secalonic acid class compound Secalonic acid M and preparation method of penicillium oxalicum - Google Patents
Derived from the secalonic acid class compound Secalonic acid M and preparation method of penicillium oxalicum Download PDFInfo
- Publication number
- CN110407798A CN110407798A CN201910345733.1A CN201910345733A CN110407798A CN 110407798 A CN110407798 A CN 110407798A CN 201910345733 A CN201910345733 A CN 201910345733A CN 110407798 A CN110407798 A CN 110407798A
- Authority
- CN
- China
- Prior art keywords
- penicillium oxalicum
- compound
- secalonic acid
- preparation
- secalonic
- 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
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/78—Ring systems having three or more relevant rings
- C07D311/80—Dibenzopyrans; Hydrogenated dibenzopyrans
- C07D311/82—Xanthenes
- C07D311/84—Xanthenes with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached in position 9
- C07D311/86—Oxygen atoms, e.g. xanthones
-
- 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
- C12P17/16—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing two or more hetero rings
- C12P17/162—Heterorings having oxygen atoms as the only ring heteroatoms, e.g. Lasalocid
Landscapes
- 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)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to a kind of secalonic acid class compound Secalonic acid M and preparation method derived from penicillium oxalicum.The compound, which has, inhibits human cancer cell proliferation function.Its structural formula are as follows:.By fermented and cultured penicillium oxalicum (Penicillium oxalicum) IBPT-6, fermentation material is obtained, the compound is then isolated and purified out from fermentation material.It is verified by experiments, which has preferable anti-tumor activity to human cancer cell.It can be used for preparing human cancer cell Proliferation Ability drug or anti-tumor drug for studying.
Description
Technical field
The present invention relates to a kind of derived from the secalonic acid class compound Secalonic acid M of penicillium oxalicum and preparation side
Method belongs to biomedicine field.
Background technique
Secalonic acid class compound (Secalonic acids) belongs to the production of ergot pigment (Ergochrome) secondary metabolism
Object is xanthone dimer.Since Stoll etc. was in the isolated secalonic acid A from fungi in 1952
After (Secalonic acid A), the compound secalonic acid (A-I) of the series is just constantly found and studies.Rye ketone
Acid compounds have a variety of different physiological activity, with secalonic acid D(Secalonic acid D, SAD) for, 5 mg/
The SAD of ml is added in physiological saline, within the scope of 5-20 mg, it can treatment early stage bladder cancer, 50-100 mg range
It is interior, and be free from side effects generation effective in cure to more serious bladder cancer.It has been investigated that some marine fungis can be in secondary
The good secalonic acid class compound of structure novel, activity is generated in metabolic process, and there is good medicinal and industrialization prospect.
The present inventor studies and learns, penicillium oxalicum (Penicillium oxalicum) IBPT-6 has been (in 2013 12
The moon is deposited in China typical culture collection center on 25th, and address: Wuhan Wuhan University, deposit number are: CCTCC NO:M
2013714) crude extract of tunning has good cell inhibitory effect activity, studies then its active constituent.It grinds
Studying carefully secalonic acid class compound shown in discovery has anticancer activity, and it is living to the Proliferation Ability of cancer cell to have not yet to see the compound
The report of property, therefore in the market also there is not yet drug related to this.
Summary of the invention
The purpose of the present invention is to provide a kind of secalonic acid class compound Secalonic acid derived from penicillium oxalicum
M and preparation method.The compound, which has, inhibits cancer cell multiplication effect, has anticancer activity.Its structural formula are as follows:
。
The preparation method of the compound, be by fermented and cultured penicillium oxalicum (Penicillium oxalicum)
IBPT-6 obtains fermentation material, the compound is then isolated and purified out from fermentation material.Specific step is as follows:
1 fermenting and producing
By culture microorganism conventional method, take penicillium oxalicum (Penicillium oxalicum) IBPT-6 is inoculated into PDA
Cultivate 2 to 3 days, be then seeded into culture solution in 28 DEG C of incubators in solid slope culture medium, 28 DEG C static gas wave refrigerator 30 days
Afterwards, mycelium and fermentation liquid are obtained；The culture solution composition: every liter of water contains 20.0 g of mannitol, 3.0 g of yeast extract, maltose
20.0 g, 10.0 g of monosodium glutamate, glucose 10.0 g, KH2PO4 0.5 g、MgSO4·7H2O 0.3 g、NaCl 15.0 g。
The acquisition of 2 medicinal extract
With gauze by mycelium and separation of fermentative broth.Mycelium with acetone soln (contain 20% ~ 30% water) continuous ultrasound broken wall 3 times,
Filtering removal residue, obtains the mycelium crude extract containing acetone and water.Removal acetone is concentrated under reduced pressure, obtains the water-soluble of crude extract
Liquid, then ethyl acetate is added with volume ratio 1:2 and is extracted 3 times, ethyl acetate crude extract is obtained, is concentrated under reduced pressure into close dry that mycelium soaks
36.5 g of cream.
The separation and purification of 3 compounds
Mycelium medicinal extract is by 100-200 mesh silica gel mixed sample, and using petroleum ether: methylene chloride: methanol is subtracted as gradient eluent
Press silica gel chromatographic column chromatography.By simple thin-layer chromatographic analysis, merges, be separated into component A-E.Component D (5.9 g) (two
Chloromethanes: the eluate of methanol v/v=100:1) using methylene chloride: methanol carries out pressured column silica gel as gradient elution agent
Analysis merges after thin-layer chromatographic analysis and obtains five subfraction D1-D5.Component D3(1.1 g) with methanol: sour water (contains 0.1%
Trifluoroacetic acid) it is that gradient elution agent carries out reversed chromatographed on silica gel, isolated four subfraction D3-1 ~ D3-4.The Asia of D3
Component D3-2(168 mg) it is separated under the conditions of mobile phase is 50% acetonitrile (containing 0.1% trifluoroacetic acid) by semi-preparative liquid chromatography
Obtain the compound (2.6mg).
The penicillium oxalicum (Penicillium oxalicum) IBPT-6, it has been deposited on December 25th, 2013
State's Type Tissue Collection, address: Wuhan Wuhan University, deposit number are: CCTCC NO:M 2013714.
The present invention also protects the compound to inhibit application and the chemical combination in human cancer cell hyperproliferation agent in preparation
Object is preparing the application in anti-human cancer drug.
Remarkable advantage of the invention: the secalonic acid compound shown in studying has not been reported and has significant inhibition people's cancer
Cell-proliferation activity has not yet to see the compound to the report of human cancer cell proliferation inhibition activity, therefore in the market also not yet
Seeing has drug related to this.
Detailed description of the invention
Fig. 1 Secalonic acid main COSY of M, HMBC and NOE signal.
Specific embodiment
The chemical structure of signified compound in the following example:
The fermenting and producing and separation and purification of 1 compound of embodiment
1 fermenting and producing
Produce bacterium fermented and cultured: by culture microorganism conventional method, take penicillium oxalicum (Penicillium oxalicum)
IBPT-6 (be deposited in China typical culture collection center on December 25th, 2013, address: protect by Wuhan Wuhan University
Hiding number is: CCTCC NO:M 2013714) in right amount, being inoculated into PDA solid slope culture medium and cultivates in 28 DEG C of incubators
2 to 3 days.
Take inclined-plane culture 2 to 3 days penicillium oxalicum (Penicillium oxalicum) appropriate IBPT-6, it is inoculated into dress
By 400mL culture solution, [culture solution is formed (grams per liter): mannitol 20.0, yeast extract 3.0, maltose 20.0, monosodium glutamate 10.0, Portugal
Grape sugar 10.0, KH2PO40.5, MgSO40.3, NaCl 15.0, constant volume] 1000mL conical flask in, 28 DEG C of static gas wave refrigerators
After 30 days, mycelium and fermentation liquid are obtained.
The acquisition of 2 medicinal extract
With gauze by mycelium and separation of fermentative broth.Mycelium with acetone soln (contain 20% ~ 30% water) continuous ultrasound broken wall 3 times,
Filtering removal residue, obtains the mycelium crude extract containing acetone and water.Removal acetone is concentrated under reduced pressure, obtains the water-soluble of crude extract
Liquid, then ethyl acetate is added with volume ratio 1:2 and is extracted 3 times, ethyl acetate crude extract is obtained, is concentrated under reduced pressure into close dry that mycelium soaks
36.5 g of cream.
The separation and purification of 3 compounds
Mycelium medicinal extract is by 100-200 mesh silica gel mixed sample, and using petroleum ether: methylene chloride: methanol is subtracted as gradient eluent
Press silica gel chromatographic column chromatography.By simple thin-layer chromatographic analysis, merges, be separated into component A-E.Component D (5.9 g) (two
Chloromethanes: the eluate of methanol v/v=100:1) using methylene chloride: methanol carries out pressured column silica gel as gradient elution agent
Analysis merges after thin-layer chromatographic analysis and obtains five subfraction D1-D5.Component D3(1.1 g) with methanol: sour water (contains 0.1%
Trifluoroacetic acid) it is that gradient elution agent carries out reversed chromatographed on silica gel, isolated four subfraction D3-1 ~ D3-4.The Asia of D3
Component D3-2(168 mg) it is separated under the conditions of mobile phase is 50% acetonitrile (containing 0.1% trifluoroacetic acid) by semi-preparative liquid chromatography
Obtain the compound (2.6mg).
Compound: being yellow oil under room temperature, and high-resolution electrospray ionization mass spectrum HRESI-MS gives at m/z:661.1535
Molecular ion peak [M+Na]+(calcd for C out32H30NaO14, 661.1533), infer that its molecular formula is C32H30O14。1H and13C-NMR data are shown in Table 1, and main COSY, HMBC and NOE signal is shown in Fig. 1.
1 compound of table1H and13C-NMR data (500MHz1H and 125 MHz 13C, in DMSO d6)
ab Signal can be interchanged.
The test of 2 anti tumor activity in vitro of embodiment
1 laboratory sample and experimental method
The preparation test sample of sample solution is the pure compounds of separation and purification in above-described embodiment 1.Precision weighs suitable
Sample is measured, the solution of required concentration is configured to methanol, for surveying activity.
The squamous subculture of cell line and cell uses tumor cell line, and tumour cell uses the DMEM containing 10% FBS to cultivate
Base, at 37 DEG C in being passed through 5% CO2Incubator in squamous subculture.
Cell inhibitory effect activity test method
Tetrazolium (MTT) method: cell density is adjusted to every milliliter 2 × 10 by the tumour cell of logarithmic growth phase5A cell,
It is inoculated in 96 porocyte culture plates for 200 microlitres by every hole, is passed through 5% CO in 37 DEG C2Incubator in cultivate 4 hours.Every hole
2 microlitres of sample liquid or blank solution is added, after culture 24 hours, MTT liquid (every milliliter of 5 milligrams of physiology of MTT are added in every hole
Saline solution) 10 microlitres, continue culture 4 hours, 37 DEG C, 2000 revs/min are centrifuged 8 minutes, draw supernatant.DMSO is added in every hole
It each 100 microlitres, is vibrated 15 minutes on micro oscillator, until utilizing the production of MD company after crystallization is completely dissolved
SPECTRAMAX Plus type microplate reader measures extinction (OD) value of every hole at 570 nm.The sample in 96 orifice plate of same
Each concentration is respectively provided with three holes, and the another blank control that three holes are arranged and cell-free withered hole are (if drug has color to do accordingly
It is withered that drug concentration is cell-free).Each hole OD value first do it is corresponding cell-free withered, then take three hole mean OD values by IR (%)=
(ODBlank control-ODSample)/ODBlank control× 100% calculates the proliferation inhibition rate (IR%) of cell under each concentration.
2. experimental result
Cell inhibitory effect active testing result
In mtt assay test, according to the Cytostatic to tumor cell rate of the compound of various concentration, using SPSS16.0 software
Carry out data processing and calculation of half inhibitory concentration IC50Value.It the results are shown in Table 2.
Inhibitory activity (the IC that 2 compound of table is proliferated human cancer cell50, μM)
3. conclusion
The compound has preferable anti-tumor activity to human cancer cell.It can be used as and prepare cancer cell multiplication inhibition drug or anti-swollen
Tumor medicine is for studying.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (3)
1. compound。
2. the preparation method of compound as described in claim 1, it is characterised in that: fermented and cultured penicillium oxalicum
(Penicillium oxalicum) IBPT-6, fermentation material is obtained, the compound is then isolated and purified out from fermentation material；Institute
State penicillium oxalicum (Penicillium oxalicum) IBPT-6, Chinese Typical Representative culture is deposited on December 25th, 2013
Object collection, address: Wuhan Wuhan University, deposit number are: CCTCC NO:M 2013714.
3. compound as described in claim 1 is preparing the application in anti-gynecological cancer drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910345733.1A CN110407798B (en) | 2019-04-26 | 2019-04-26 | Secalonic acid M compound derived from penicillium oxalicum and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910345733.1A CN110407798B (en) | 2019-04-26 | 2019-04-26 | Secalonic acid M compound derived from penicillium oxalicum and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110407798A true CN110407798A (en) | 2019-11-05 |
| CN110407798B CN110407798B (en) | 2023-04-07 |
Family
ID=68358161
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910345733.1A Active CN110407798B (en) | 2019-04-26 | 2019-04-26 | Secalonic acid M compound derived from penicillium oxalicum and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110407798B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4424373A (en) * | 1979-01-30 | 1984-01-03 | Asahi Kasei Kogyo Kabushiki Kaisha | Secalonic acids |
| JP2002047181A (en) * | 2000-08-07 | 2002-02-12 | Japan Research Association For Reforestation Of Tropical Forest | Agent for suppressing production of nitric oxide |
| CN109106702A (en) * | 2017-12-19 | 2019-01-01 | 福州大学 | Derived from application of 4-4 ' the isomerization secalonic acid D in terms of colon cancer of penicillium oxalicum |
-
2019
- 2019-04-26 CN CN201910345733.1A patent/CN110407798B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4424373A (en) * | 1979-01-30 | 1984-01-03 | Asahi Kasei Kogyo Kabushiki Kaisha | Secalonic acids |
| JP2002047181A (en) * | 2000-08-07 | 2002-02-12 | Japan Research Association For Reforestation Of Tropical Forest | Agent for suppressing production of nitric oxide |
| CN109106702A (en) * | 2017-12-19 | 2019-01-01 | 福州大学 | Derived from application of 4-4 ' the isomerization secalonic acid D in terms of colon cancer of penicillium oxalicum |
Non-Patent Citations (5)
| Title |
|---|
| DUN-MEI YANG ET AL.: ""Structures of eumitrins A1, A2, and B, yellow pigments of the lichen, Usnea bayleyi"", 《TETRAHEDRON》 * |
| LI CHEN ET AL.: ""Secalonic acids J-M, four new secondary metabolites from the marine-derived fungus Penicillium oxalicum"", 《HETEROCYCLES》 * |
| MAULIDIYAH ET AL.: ""Isolation and structure elucidation of Eumitrin A1 from lichen Usnea blepharea motyka and its cytotoxic activity"", 《INTERNATIONAL JOURNAL OF PHARMTECH RESEARCH》 * |
| WEN ZHANG ET AL.: ""New Mono- and Dimeric Members of the Secalonic Acid Family: Blennolides A–G Isolated from the Fungus Blennoria sp."", 《CHEM. EUR. J.》 * |
| 李庆 等: ""真菌代谢产物Secalonic acid D及其药理活性研究"", 《中药材》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110407798B (en) | 2023-04-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107298671B (en) | Selenolonic acid H from penicillium oxalicum and application thereof in preparing medicine for resisting human colon cancer | |
| CN107353274B (en) | Selenolonic acid I from penicillium oxalicum and application thereof in preparation of human esophageal cancer resistant medicine | |
| CN107298672B (en) | Application of seclenic acid I derived from penicillium oxalicum in preparation of anti-human colon cancer drugs | |
| CN107298670B (en) | Application of medicine derived from penicillium oxalicum seclenum ketonic acid H in preparation of anti-human oral epidermoid carcinoma medicines | |
| CN107485607B (en) | Application of seclenic acid H derived from penicillium oxalicum in preparation of human esophageal cancer resistant medicine | |
| CN107298669B (en) | Selenolonic acid I from penicillium oxalicum and application of medicine for resisting human oral epidermoid carcinoma | |
| CN110407792A (en) | Derived from the secalonic acid class compound Secalonic acid J and preparation method of penicillium oxalicum | |
| CN109106702A (en) | Derived from application of 4-4 ' the isomerization secalonic acid D in terms of colon cancer of penicillium oxalicum | |
| CN109134416A (en) | Secalonic acid H derived from penicillium oxalicum is in the application for preparing human cervical cancer 1 cancer drug | |
| CN110407797A (en) | Derived from the secalonic acid class compound Secalonic acid K and preparation method of penicillium oxalicum | |
| CN110407794A (en) | Secalonic acid K derived from penicillium oxalicum and the application on inhibition cancer cell multiplication | |
| CN109776477A (en) | Iso-Penicillixanthone A and anti-vincristine drug resistance application derived from penicillium oxalicum | |
| CN109134417A (en) | Derived from the application of the secalonic acid I and anti-human cervix cancer drug of penicillium oxalicum | |
| CN110407798A (en) | Derived from the secalonic acid class compound Secalonic acid M and preparation method of penicillium oxalicum | |
| CN110403929A (en) | Derived from the secalonic acid M of penicillium oxalicum and in the application for inhibiting human cancer cell to be proliferated | |
| CN110407793A (en) | Secalonic acid J derived from penicillium oxalicum and the application on inhibition cancer cell multiplication | |
| CN110407795A (en) | Derived from penicillium oxalicum secalonic acid L and inhibiting the application in human cancer cell proliferation | |
| CN110407796A (en) | Derived from the secalonic acid class compound Secalonic acid L and preparation method of penicillium oxalicum | |
| CN106491596B (en) | Derived from application of the mould enol E1 in terms of lung cancer of tangerine green trichoderma | |
| CN106432035A (en) | Application of penem enol E1 derived from trichoderma citrinoviride on the aspect of malignant melanoma | |
| CN106389418B (en) | Derived from tangerine green trichoderma mould enol E1 liver cancer application | |
| CN105131006B (en) | Penicitrinine A sourced from penicillium citrinum and application thereof in preparation of anti-malignant melanoma drug | |
| CN105061443B (en) | Penicillium citrinum-derived penicitrinine A as well as application thereof to preparation of drugs for resisting human liver cancer | |
| CN109106703A (en) | Derived from application of 4-4 ' the isomerization secalonic acid D in terms of nasopharyngeal carcinoma of penicillium oxalicum | |
| CN109106701A (en) | Derived from application of 4-4 ' the isomerization secalonic acid D in terms of lymthoma of penicillium oxalicum |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |