CN106420716B - Derived from application of the mould enol E1 in terms of intestinal cancer of tangerine green trichoderma - Google Patents
Derived from application of the mould enol E1 in terms of intestinal cancer of tangerine green trichoderma Download PDFInfo
- Publication number
- CN106420716B CN106420716B CN201610820509.XA CN201610820509A CN106420716B CN 106420716 B CN106420716 B CN 106420716B CN 201610820509 A CN201610820509 A CN 201610820509A CN 106420716 B CN106420716 B CN 106420716B
- Authority
- CN
- China
- Prior art keywords
- trichoderma
- derived
- green trichoderma
- tangerine green
- enol
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/4015—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil having oxo groups directly attached to the heterocyclic ring, e.g. piracetam, ethosuximide
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/30—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having two double bonds between ring members or between ring members and non-ring members
- C07D207/34—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having two double bonds between ring members or between ring members and non-ring members 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 to ring carbon atoms
- C07D207/36—Oxygen or sulfur atoms
- C07D207/38—2-Pyrrolones
-
- 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
- C12P17/10—Nitrogen as only ring hetero atom
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The application that the present invention relates to a kind of derived from the mould enol E1 of tangerine green trichoderma in terms of intestinal cancer.The compound structure is characterized in: the molecular skeleton containing five cyclic amides, containing connecting on one ten carbon saturated fat long-chain of a carbonyl connection, N, a methyl, there are two hydroxyl groups for molecule.It is verified by experiments, the alkaloid compound has preferable inhibitory activity to colon-cancer cell, can be used as and prepares colon-cancer cell Proliferation Ability drug or anti-bowelcancer medicine for antitumor research.
Description
Technical field
The present invention relates to a kind of mould enol E1 derived from tangerine green trichoderma to apply in terms of intestinal cancer.
Background technique
Alkaloid is a kind of organic compounds containing nitrogen generated by biological cometabolism, the alkaloid type in nature compared with
It is more, greatly mostly from plant, therefore there is the title of vegetable soda again.Alkaloid has important physiological action to humans and animals, including flat
Antibechic, hypoglycemic, reducing blood lipid, antibacterial, antitumor, analgesia etc. are breathed heavily, wherein the most prominent with antibacterial, anti-tumor activity.Natural knot
Structure alkaloid is the important sources that lead compound is found in innovation drug research, has been applied to clinical alkaloidal drug at present
Through nearly hundred kinds.The study found that some marine fungis can generate structure novel, activity good biology during cometabolism
Alkali has good medicinal and industrialization prospect.
The present inventor studies and learns, tangerine green trichoderma (Trichoderma citrinoviride.) IBPT-4 has been (in 2013
On January 25, in is deposited in China typical culture collection center, and address: Wuhan Wuhan University, deposit number are: CCTCC
NO:M 2013055) the crude extract of tunning have good tumor cell proliferation inhibition activity, then to its active constituent
It is studied, tangerine green trichoderma (Trichoderma citrinoviride.) IBPT-4 is in the patent No. 201310208563.5
It discloses.Research finds that shown alkaloid compound has anti-tumor activity for intestinal cancer, has not yet to see the compound
Chemical structure and report for colon-cancer cell proliferation inhibition activity, therefore in the market also there is not yet drug related to this.
Summary of the invention
The application that the purpose of the present invention is to provide a kind of derived from the mould enol E1 of tangerine green trichoderma in terms of intestinal cancer.
Present invention firstly relates to one plant of tangerine green trichoderma (Trichoderma citrinoviride.) IBPT-4, the bacterial strain
It is deposited in China typical culture collection center on January 25th, 2013, address: Wuhan Wuhan University, deposit number are:
CCTCC NO:M 2013055；Tangerine green trichoderma (Trichoderma citrinoviride.) IBPT-4 is in the patent No.
It is disclosed in 201310208563.5.
The purposes of the bacterial strain is, by tangerine green trichoderma (Trichoderma citrinoviride.) IBPT-4 sent out
Ferment culture, mycelium and fermentation broth extract are isolated to the compound mould enol with tumor cell proliferation inhibition activity
E1。
The structural formula of compound are as follows:
。
Its structure feature is: the molecular skeleton containing five cyclic amides connects a ten carbon saturation containing a carbonyl
On fatty long-chain, N connect a methyl, there are two hydroxyl groups for molecule.
The present invention also protects the compound in preparation for the application in colon-cancer cell Proliferation Ability drug, and should
Compound is preparing the application in anti-bowelcancer medicine.
Remarkable advantage of the invention: the compound shown in studying is the alkaloid of a structure novel, the alkaloids
Compound has significant anti-intestinal cancer activity, has not yet to see the chemical structure of the compound and for colon-cancer cell Proliferation Ability
Active report, therefore in the market also there is not yet drug related to this.
Detailed description of the invention
Fig. 1 is the main COSY and HMBC signal of mould enol E1.
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 tangerine green trichoderma (Trichoderma citrinoviride.) IBPT-4 (is deposited in China typical culture collection center on January 25th, 2013, address: military
Chinese Wuhan University, deposit number are: CCTCC NO:M 2013055) in right amount, it is inoculated into PDA solid slope culture medium 28
It is cultivated 4 days in DEG C incubator.
Take inclined-plane culture 4 days tangerine green trichoderma (Trichoderma citrinoviride.) appropriate IBPT-4, it is inoculated into
Equipped with 400mL culture solution [culture solution form (grams per liter): mannitol 20.0, yeast extract 3.0, maltose 20.0, monosodium glutamate 10.0,
Glucose 10.0, KH2PO40.5, MgSO40.3, NaCL 6.0 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.Fermentation liquid and ethyl acetate 1:2(v/v) are extracted twice, extract liquor
Vacuum distillation obtains the ethyl acetate extract of fermentation liquid to doing.Mycelium is then with the aqueous solution ultrasonication of the acetone containing 70%-80%
It 3 times, removes and removal acetone is concentrated under reduced pressure after residue merges clear liquid, be extracted twice with ethyl acetate is added for 1:2 by volume,
It is concentrated to dryness to obtain mycelium medicinal extract.The total medicinal extract of extract is obtained after mycelium and fermentation liquid medicinal extract are merged.
The separation and purification of 3 compounds
After the medicinal extract passes through 100-200 mesh silica gel mixed sample, using petroleum ether: methylene chloride: methanol depressurizes silicon as eluent
Glue chromatographic column gradient elution.Eluent obtains active component B (methylene chloride-methanol v/v 50:1 elution by activity tracking
Object), then with petroleum ether: methylene chloride: methanol gradient group is divided into eluant, eluent, is further washed by pressurized silica gel column chromatographic grade
It is de-, obtained active subfraction B1 (eluate that methylene chloride-methanol is 20:1) with chloroform-methanol (v/v1:2) be solvent into
Row Sephadex LH-20 gel filtration chromatography, finally by semi-preparative liquid chromatography (1010 type ODS-A, 10 × 250 mm, 5 μ
M): separation flow velocity be 5 mL/min, mobile phase be 85% acetonitrile contain 0.1% TFA, obtain shown compound (32.1 mg,t R
20.7min).
Compound light yellow oil, high resolution mass spectrum HRESI-MS existm/z324.2161 place provides molecular ion peak
[M – H]–, (Calcd for C18H30NO4, 324.2175), prompting molecular weight is 325, speculates molecular formula in conjunction with spectral information
For C18H31NO4。 1H and13The NMR datas such as C-NMR are shown in Table 1.
1 compound of table1H and13C-NMR data (500 MHz, in CDCl3)a)
A) this table signals assignment is based on DEPT, HMQC and HMBC spectrum analysis result.The type of carbon signal utilizes the side DEPT
Method determines.
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-mentioned implementation 1.Precision weighs
Appropriate amount of sample is configured to the solution of required concentration with methanol, for surveying activity.
The squamous subculture of cell line and cell uses colon-cancer cell system, and cell uses the RPMI-1640 containing 10% FBS to cultivate
Base, in 37 DEG C of squamous subcultures in the incubator for being passed through 5% carbon dioxide.
Cell inhibitory effect activity test method
The cell of tetrazolium (MTT) method logarithmic growth phase, is adjusted to every milliliter 2 × 10 for cell density5A 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 liquids or blank solution is added, after culture 24 hours, every hole adds MTT liquid, and (every milliliter of 5 milligrams of physiology salts of MTT are water-soluble
Liquid) 10 microlitres, continue culture 4 hours, 37 DEG C, 2000 revs/min are centrifuged 8 minutes, suck supernatant.DMSO each 100 is added in every hole
Microlitre, it is vibrated 15 minutes on micro oscillator, until producing SPECTRAMAX Plus type using MD company after crystallization is completely dissolved
Microplate reader measures light absorption value (OD) value of every hole at 570nm.Each concentration of sample is respectively provided with three in 96 orifice plate of same
Hole separately sets three hole blank controls and cell-free zeroing hole (if drug has color to do the cell-free zeroing of relative medicine concentration).
Each hole OD value first does corresponding cell-free zeroing, then takes three hole mean OD values by IR (%)=(ODBlank control-ODSample)/ODBlank control ×
100% formula calculates cell proliferation inhibition rate (IR%) under each concentration.
2. experimental result
Cell inhibitory effect active testing result
In mtt assay test, according to the colon-cancer cell proliferation inhibition rate of the compound of various concentration, using SPSS16.0
Software carries out data processing and calculation of half inhibitory concentration IC50Value.It the results are shown in Table 2.
The inhibitory activity that 2 compound of table is proliferated colon-cancer cell
3. conclusion
Compound has apparent colon-cancer cell inhibited proliferation, can be used as and prepares colon-cancer cell antiblastic or anti-
Bowelcancer medicine is used for antitumor research.
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 (2)
1. a kind of application of the mould enol E1 derived from tangerine green trichoderma, it is characterised in that: the structural formula of the mould enol E1 is, preparation of the mould enol E1 applied to colon-cancer cell Proliferation Ability drug；
The mould enol E1 be by tangerine green trichoderma (Trichoderma citrinoviride) IBPT-4 progress fermented and cultured, then
It is isolated from its mycelium and fermentation broth extract.
2. a kind of application of the mould enol E1 derived from tangerine green trichoderma, it is characterised in that: the structural formula of the mould enol E1 is, preparation of the mould enol E1 applied to anti-bowelcancer medicine；The penem
Alcohol E1 be by tangerine green trichoderma (Trichoderma citrinoviride) IBPT-4 progress fermented and cultured, then from its mycelium
With it is isolated in fermentation broth extract.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610820509.XA CN106420716B (en) | 2016-09-13 | 2016-09-13 | Derived from application of the mould enol E1 in terms of intestinal cancer of tangerine green trichoderma |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610820509.XA CN106420716B (en) | 2016-09-13 | 2016-09-13 | Derived from application of the mould enol E1 in terms of intestinal cancer of tangerine green trichoderma |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106420716A CN106420716A (en) | 2017-02-22 |
| CN106420716B true CN106420716B (en) | 2019-06-07 |
Family
ID=58168961
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610820509.XA Expired - Fee Related CN106420716B (en) | 2016-09-13 | 2016-09-13 | Derived from application of the mould enol E1 in terms of intestinal cancer of tangerine green trichoderma |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106420716B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103865808A (en) * | 2014-03-10 | 2014-06-18 | 福州大学 | Novel anti-tumor application of penicillium enol A1 from penicillium citrinum |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110136752A1 (en) * | 2009-12-04 | 2011-06-09 | Novobiotic Pharmaceuticals Llc | Novel antibiotics |
-
2016
- 2016-09-13 CN CN201610820509.XA patent/CN106420716B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103865808A (en) * | 2014-03-10 | 2014-06-18 | 福州大学 | Novel anti-tumor application of penicillium enol A1 from penicillium citrinum |
Non-Patent Citations (2)
| Title |
|---|
| Penicillenols from Penicillium sp. GQ-7, an Endophytic Fungus Associated with Aegiceras corniculatum;Zhen-Jian LIN等;《Chem. Pharm. Bull.》;20071112;第56卷(第2期);第217-221页，尤其是第217页左栏第2段，第220页右栏第6段，图1，表3 |
| TUMONOIC ACIDS K AND L, NOVEL METABOLITES FROM THE MARINE-DERIVED FUNGUS PENICILLIUM CITRINUM;Li Chen等;《HETEROCYCLES》;20111208;第85卷(第2期);第413-419页，尤其是第414页图1，416页6-8行 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106420716A (en) | 2017-02-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103865809B (en) | A kind of anticancer usage of the penicillium sp enol B1 coming from Aspergillus citrimum | |
| CN104592082B (en) | Come from the penicillium sp enol D of Aspergillus citrimum2preparation method and applications | |
| CN105061446B (en) | Penicillium citrinum-derived penicitrinine A as well as application thereof to preparation of drugs for resisting nasopharyngeal carcinoma | |
| CN106420716B (en) | Derived from application of the mould enol E1 in terms of intestinal cancer of tangerine green trichoderma | |
| CN106420715B (en) | Mould enol E1 derived from tangerine green trichoderma is in the application for preparing anti-lymphadenoma drug | |
| CN106420717B (en) | Derived from application of the mould enol E1 in terms of nasopharyngeal carcinoma of tangerine green trichoderma | |
| CN106432034B (en) | Mould enol E1 derived from tangerine green trichoderma is in the application for preparing anti-breast cancer medicines | |
| CN106432036B (en) | Derived from tangerine green trichoderma mould enol E1 and prepare anti-oral cavity epidermal carcinoma medicinal application | |
| CN106491597B (en) | Derived from application of the mould enol E1 in terms of the cancer of the esophagus of tangerine green trichoderma | |
| CN106389417B (en) | Derived from application of the mould enol E1 in terms of gastric cancer of tangerine green trichoderma | |
| CN106491596B (en) | Derived from application of the mould enol E1 in terms of lung cancer of tangerine green trichoderma | |
| CN106389418B (en) | Derived from tangerine green trichoderma mould enol E1 liver cancer application | |
| CN104447475B (en) | Preparation method and application of penicillenol D1 derived from penicillium citrinum | |
| CN107325087B (en) | Citrinin compounds dicitrinone D and its application in terms of malignant mela noma | |
| CN110407797A (en) | Derived from the secalonic acid class compound Secalonic acid K and preparation method of penicillium oxalicum | |
| CN103265522B (en) | Lactone derivative derived from trichoderma citrinoviride and application thereof | |
| CN106432035A (en) | Application of penem enol E1 derived from trichoderma citrinoviride on the aspect of malignant melanoma | |
| CN104211780A (en) | Cyclic depsipeptides compound, preparation method and purpose thereof | |
| CN105061443B (en) | Penicillium citrinum-derived penicitrinine A as well as application thereof to preparation of drugs for resisting human liver cancer | |
| CN105131006B (en) | Penicitrinine A sourced from penicillium citrinum and application thereof in preparation of anti-malignant melanoma drug | |
| CN107325085B (en) | Citrinin compounds dicitrinone D preparation method and its application in terms of lymph cancer | |
| CN105061445B (en) | Penicillium citrinum-derived penicitrinine A as well as application thereof to preparation of drugs for resisting human gastric cancer | |
| CN104211630B (en) | A kind of alkaloid compound and application thereof that comes from tangerine green trichoderma | |
| CN107325088B (en) | Citrinin compounds dicitrinone D preparation method and the application in terms of nasopharyngeal carcinoma | |
| CN105153175B (en) | The penicitrinine A for coming from Penicillium citrinum and its application for preparing anti-human oesophagus cancer drug |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190607 Termination date: 20210913 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |