CN104293846B - A kind of preparation method of butyrolactone compound and the application as marine antifoulant - Google Patents
A kind of preparation method of butyrolactone compound and the application as marine antifoulant Download PDFInfo
- Publication number
- CN104293846B CN104293846B CN201410512799.2A CN201410512799A CN104293846B CN 104293846 B CN104293846 B CN 104293846B CN 201410512799 A CN201410512799 A CN 201410512799A CN 104293846 B CN104293846 B CN 104293846B
- Authority
- CN
- China
- Prior art keywords
- compound
- marine
- formula
- preparation
- ethyl acetate
- 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.)
- Active
Links
Abstract
A kind of preparation method of butyrolactone compound and the application as marine antifoulant, Spawn incubation first is carried out to fungi Aspergillus terreus (WZ 2,011 2905) during preparation, fermented and cultured is carried out to the fungi again, gained mycelium chloroform methanol mixed liquor (1: 1) is extracted 3 times after being concentrated under reduced pressure, coarse extract is extracted with ethyl acetate 3 times to obtain；Ethyl acetate coarse extract carries out normal-phase silica gel column chromatography, the gel filtration chromatographies of Sephadex LH 20, HPLC high performance liquid chromatography successively, produces compound of formula I.The present invention provides a kind of marine organisms anti-fouling agent, it is characterised in that with the compound of formula I or its pharmaceutically acceptable salt of the present invention, for preventing and treating marine biofouling caused by barnacle attachment.
Description
Technical field
The present invention relates to a kind of preparation method of butyrolactone (butyrolactones) compound and application, more particularly to
A kind of butyrolactone and its system to marine fouling organism barnacle Balanus amphitrite larvas with extremely strong inhibitory activity
Preparation Method and application.
Background technology
Marine biofouling is that subduction is set in ocean for organic molecule, microorganism, animal, plant and their accessory substance
Apply the harmfulness accumulation on surface.This harmfulness accumulation frequently occurs in the surface for the ocean subduction facility do not protected, including
Sea-freight and the ship of tourism, naval's warship, heat exchanger, sea sensor and aquaculture base etc..Biodeterioration causes
Huge economic loss, only by taking USN's warship as an example, every year economic loss in this respect 18 to 26 hundred million dollars it
Between, and USN's warship quantity only accounts for the 0.5% of global ships quantity, therefore marine biofouling is extremely serious nature
Harm.Barnacle is representativeness biology very universal in fouling organism known today because of its very strong Adhering capacity.From 2008
After the year global use for eliminating poisonous anti-fouling agent organotin, finding safe and efficient marine antifoulant turns into urgent need solution in the world
Problem certainly.Marine natural products is considered as the important sources of novel sea anti-fouling agent.In fact, in the past few decades
The compound for much having strong anti-fouling activity is found that from sponge, the marine organisms such as coral and marine alga.However, from above-mentioned
The reactive compound found in macro-organism is due to being leveraged its potential application by the limitation measured.Marine microorganism
Due in the lab can with large scale fermentation, survivable natural environment, and as activity marine compound it is most important come
Source.However, in recent years there is not yet the butyrolactone compound that important anti-fouling activity is obtained from the fungi in sea hare source is made
For the use of anti-fouling agent.(J.A.Callow, M.E.Callow, Nat.Commun.2011,2,244-253；
C.M.Kirschner, A.B.Brennan, Annu.Rev.Mater.Res.2012,42,1-19；M.Schultz,
J.Bendick, E.Holm, W.Hertel, Biofouling 2011,27,87-98；N.Fusetani,
Nat.Prod.Rep.2004,21,94-104；N.Fusetani, Nat.Prod.Rep.2011,28,400-410；P.-
Y.Qian, Y.Xu, N.Fusetani, Biofouling 2010,26,223-234.)
The content of the invention
It is an object of the invention to provide a kind of preparation method of butyrolactone compound from aplysiatoxin fungi and conduct
The application of marine antifoulant, it can meet the demand of prior art.Culture presevation information：Depositary institution's title：China is micro-
Biological inoculum preservation administration committee common micro-organisms center；Depositary institution address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number Institute of Microorganism, Academia Sinica；Preservation date：On 09 02nd, 2014；Deposit number：CGMCC9517；Classification And Nomenclature：
Aspergillus terreus。
The present invention provides compound of formula I or its pharmaceutically acceptable salt：Or its medicine
Acceptable salt on.
Compound of formula I is selected from following compounds：
The present invention provides the preparation method of compound of formula I, it is characterised in that first to fungi in bacterium culture medium
Aspergillus terreus (WZ-2011-2905) carry out Spawn incubation, then the fungi is sent out in the fermentation medium
Ferment culture, gained mycelium chloroform-methanol mixed liquor (1: 1) is extracted 3 times after being concentrated under reduced pressure, be extracted with ethyl acetate 3 times
Obtain coarse extract；After ethyl acetate coarse extract carries out normal-phase silica gel column chromatography, Sephadex LH-20 gel filtration chromatographies respectively, then
Through HPLC high performance liquid preparative chromatographies, gained eluent is concentrated, as compound of formula I.
Bacterium culture medium preferably comprises glucose 0.1%-5.0% (percentage by weight, similarly hereinafter), ferment in above-mentioned preparation method
Female cream 0.01%-1%, peptone 0.01%-1%, agar 0.1%-3.0%, sodium chloride 0.05%-5%, remaining is water, training
Foster temperature is preferably 0-30 DEG C, and incubation time is preferably 3-15 days；Fermentation medium preferably comprises rice 1.0%-80.0% (weights
Measure percentage, similarly hereinafter), sodium chloride 0.05%-5%, remaining is water, and cultivation temperature is preferably 0-30 DEG C, and incubation time is preferably
10-60 days；The preferred 200-300 mesh silica gel of stationary phase that described normal-phase silica gel column chromatography uses, mobile phase preferred volume ratio are
15%-60% ethyl acetate-light petrol mixed solvent；The mobile phase that the Sephadex LH-20 gel filtration chromatographies use is excellent
It is chloroform: methanol=1: 1 mixed solvent to select volume ratio；The chromatographic column used in the HPLC high performance liquid preparative chromatographies is this
Field routine ODS C18 posts, preferably 10 × 250mm of Kromasil, 5 μm, flow velocity is preferably 1.0-5.0mL/min, mobile phase
Preferred volume ratio is 50%-80% Methanol+Water.
The butyrolactone compound that the present invention obtains from aplysiatoxin fungi is to marine fouling organism barnacle Balanus
Amphitrite larvas have extremely strong inhibitory activity, available for marine antifoulant is developed, have a extensive future.
Another embodiment of the present invention provides compound of formula I or its pharmaceutically acceptable salt is preparing marine antifoulant
In application.
Term " pharmaceutically acceptable salt " refers to the addition of atoxic inorganic or organic acid and/or alkali in the present invention
Salt.Reference can be made to " Salt selection for basic drugs ", Int.J.Pharm. (1986), 33,201-217.
Embodiment
For the ease of a further understanding of the present invention, examples provided below has done more detailed description to it.But
It is that these embodiments only are not used for limiting the scope of the present invention or implementation principle, reality of the invention for being better understood from inventing
The mode of applying is not limited to herein below.
Embodiment 1
(1) sea hare source fungi Aspergillus terreus (WZ-2011-2905) Spawn incubation
Culture medium used in Spawn incubation contains glucose 1.0% (percentage by weight, similarly hereinafter), yeast extract 0.2%, albumen
Peptone 0.2%, agar 1.0%, sodium chloride 3.0%, remaining is water, test tube slant is made during use, fungal bacterial strain is trained at 30 DEG C
Support 3 days.
(2) sea hare source fungi Aspergillus terreus (WZ-2011-2905) fermentation
Culture medium used in fermented and cultured contains rice 40.0% (percentage by weight, similarly hereinafter), sodium chloride 3.0%, remaining
For water；Fungal bacterial strain is cultivated 30 days in 28 DEG C.
(3) the extraction separation of compound of formula I
The mycelium for taking 80 bottles of steps (2) to obtain, extracted 3 times after being concentrated under reduced pressure, used with chloroform-methanol mixed liquor (1: 1)
Ethyl acetate extracts 3 times to obtain coarse extract；Ethyl acetate coarse extract carries out normal-phase silica gel column chromatography (stationary phase 200- respectively
300 mesh silica gel；Mobile phase is 70% ethyl acetate/petroleum ether mixed solvent, volume ratio), Sephadex LH-20 gel column layers
After analysing (chloroform: methanol=1: 1 mixed solvent, volume ratio), then (chromatographic column is through the separation of HPLC high performance liquid preparative chromatographies
Kromasil 10 × 250mm, 5 μm, flow velocity 2.0mL/min, mobile phase is 90% Methanol+Water, volume ratio),
Gained eluent is concentrated, obtains colourless crystallization, as compound of formula I.
The structural identification data of compound of formula I：
Brown oil；ES1-MSm/z 425.1[M+H]+, 447.1 [M+Na]+；Proton nmr spectra1H-NMR(CD3OD,
600MHz) 3.76 (3H, s, MeO-5), 3.44 (2H, dd, J=14.4,15Hz, H-6aand H-6b), 6.42 (1H, d, J=
2.4Hz, H-2 "), 6.50 (1H, d, J=8.4Hz, H-5 "), 6.54 (1H, dd, J=8.4,2.4Hz, H-6 "), 3.08 (2H, m,
H-7 "), 5.07 (1H, br t, J=7.2Hz, H-8 "), 1.66 (3H, s, H3- 10 "), 1.57 (3H, s, H3- 11 "), 7.60 (2H,
D, J=9.0Hz, H-2 ' and H-6 '), 6.88 (2H, d, J=9.0Hz, H-3 ' andH-5 ')；Carbon-13 nmr spectra 13C-NMR
(CD3OD, 600MHz) 170.5 (C, C-1), 133.0 (C, C-2), 129.2 (C, C-3), 86.8 (C, C-4), 171.6 (C, C-
5), 53.9 (CH3, MeO-5), 39.6 (CH2, C-6), 123.5 (C, C-1 "), 129.8 (CH, C-2 "), 116.6 (CH, C-3 "),
155.0 (C, C-4 "), 129.8 (CH, C-5 "), 116.6 (CH, C-6 "), 28.7 (CH2, C-7 "), 123.2 (CH, C-8 "),
132.4 (C, C-9 "), 26.0 (CH3, C-10 "), 17.8 (CH3, C-11 "), 125.1 (C, C-1 '), 130.4 (CH, C-2 '),
128.4 (C, C-3 '), 159.2 (C, C-4 '), 115.0 (CH, C-5 '), 130.4 (CH, C-6 ').
Embodiment 2
(1) sea hare source fungi Aspergillus terreus (WZ-2011-2905) Spawn incubation
Culture medium used in Spawn incubation contains glucose 0.1%-5.0% (percentage by weight, similarly hereinafter), yeast extract
0.01%-1%, peptone 0.01%-1%, agar 0.1%-3.0%, sodium chloride 0.05%-5%, remaining is water, during use
Test tube slant is made, fungal bacterial strain is cultivated 3-15 days at 0-30 DEG C.
(2) sea hare source fungi Aspergillus terreus (WZ-2011-2905) fermentation
Culture medium used in fermented and cultured contains rice 1.0%-80.0% (percentage by weight, similarly hereinafter), sodium chloride
0.05%-5%, remaining is water, and fungal bacterial strain is cultivated 10-60 days in 0-30 DEG C.
(3) the extraction separation of compound of formula I
Take obtained by 10-60 bottles step (2) that gained mycelium chloroform-methanol mixed liquor (1: 1) 3 decompressions of extraction is dense
After contracting, coarse extract is extracted with ethyl acetate 3 times to obtain；Normal-phase silica gel column chromatography is carried out respectively after the concentration of ethyl acetate coarse extract
(stationary phase is this area routine purification on normal-phase silica gel, and mobile phase is 15%-40% ethyl acetate-light petrol mixed solvent, volume
Than), after Sephadex LH-20 gel filtration chromatographies (mobile phase is chloroform: methanol=1: 1 mixed solvent, volume ratio), then pass through
(chromatographic column is this area routine ODS C18 posts to HPLC high performance liquid preparative chromatographies, flow velocity 1.0-5.0mL/min, and mobile phase is
50%-80% Methanol+Water, volume ratio), gained eluent is concentrated, obtains colourless crystallization, as compound of formula I.
The structural identification data of its compounds of formula I is consistent with corresponding data in embodiment 1.
Other Spawn incubations, the fermentation condition not particularly pointed out in embodiment 1-2, and normal phase silica gel column chromatography separation,
Other experimental operating conditions such as the separation of Sephadex LH-20 gel filtration chromatographies, high performance liquid chromatography separation are this area routine
Experimental operating conditions, those skilled in the art can reasonably be selected according to being actually needed.
Embodiment 3
Strain is carried out to sea hare source fungi Aspergillus terreus (WZ-2011-2905) in bacterium culture medium
Culture, then fermented and cultured is carried out to the fungi in the fermentation medium, by gained filtering fermentation liquor, thalline is removed, filtrate is dense
After contracting, it is extracted with ethyl acetate；Normal-phase silica gel column chromatography, Sephadex LH-20 gel column layers are carried out respectively after extract concentration
After analysis, then through HPLC high performance liquid preparative chromatographies, gained eluent is concentrated, obtains clear crystal, as compound of formula I, it is tied
Structure confirmation data are consistent with corresponding data in embodiment 1.Contain glucose, yeast extract, albumen in wherein described bacterium culture medium
Peptone, agar, coarse sea salt, water, rice, coarse sea salt, peptone, water are contained in the fermentation medium；Described chromatographic isolation is
Normal phase silica gel column chromatography separation, Sephadex LH-20 gel filtration chromatographies and high performance liquid chromatography separation.
In order to explore the method for being widely applied to prepare formula I, bacterium culture medium in the present embodiment,
The addition of each composition in fermentation medium, add in conventional ratio in this area or add in any proportion, during chromatographic isolation
The selection of the specification of used silica gel, the specification of Sephadex LH-20 gels, the model of chromatographic column and eluting solvent, is ability
The conventional selection in domain.Test result indicates that the preparation method of above-mentioned conventional selection, the clear crystal that can be invented, i.e. Formulas I
Compound, its structural identification data is consistent with corresponding data in embodiment 1, only exists individual compound in purity and yield side
The fine difference in face.
Embodiment 1-3 result shows, according to the conventional Spawn incubation in this area, fermentation condition, conventional purification on normal-phase silica gel
Pillar layer separation, the separation of Sephadex LH-20 gel column chromatographies, the condition of high performance liquid chromatography separation are to sea hare source fungi
Aspergillus terreus (WZ-2011-2905) carry out Spawn incubation, ferment, isolate and purify, and can obtain formula I
The compound of structure.The preparation method of formula I, the method described in preferred embodiment 1-2.
Embodiment 4
The compound of formula I of the present invention is tested according to following document barnacle Balanus amphitrite larvas attachment activity
Method is tested：Thiyagarajan, V.；Harder, T.；Qiu, J.W.；Qian, P.Y.Mar.Biol. (Berlin) 2003,
143,543-554.
The compound of formula I of the present invention and its attachment of crystal barnacle B.amphitrite larvas have extremely strong inhibitory activity,
Its EC50It is worth for 0.63 μ g/mL, and there is very high security, its toxicity efficiency ratio LC50/EC50Value is more than 79.Above-mentioned activity
Much it is better than potential natural anti-fouling compound EC as defined in USN50The μ g/mL of value 25 standard.Importantly, it
Toxicity efficiency ratio (LC50/EC50) much larger than 15, and LC50/EC50It is regarded as that there is exploitation more than 15 into safety antifouling agent
Potentiality.This shows compound of formula I or its pharmaceutically acceptable salt available for the marine antifoulant for preparing high-efficiency low-toxicity, and
Sea hare source fungi Aspergillus terreus (WZ-2011-2905) can carry out large scale fermentation production, ensure that Formulas I
The natural origin of compound, has broad application prospects.
Claims (1)
1. a kind of butyrolactone compound of Formulas I structure or its pharmaceutically acceptable salt are in anti-barnacle marine antifoulant is prepared
Using,
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410512799.2A CN104293846B (en) | 2014-09-20 | 2014-09-20 | A kind of preparation method of butyrolactone compound and the application as marine antifoulant |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410512799.2A CN104293846B (en) | 2014-09-20 | 2014-09-20 | A kind of preparation method of butyrolactone compound and the application as marine antifoulant |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104293846A CN104293846A (en) | 2015-01-21 |
| CN104293846B true CN104293846B (en) | 2018-01-16 |
Family
ID=52313794
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410512799.2A Active CN104293846B (en) | 2014-09-20 | 2014-09-20 | A kind of preparation method of butyrolactone compound and the application as marine antifoulant |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104293846B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434372B (en) * | 2016-09-29 | 2020-07-03 | 广东海洋大学 | Application of coral-derived fungus aspergillus terreus strain C21-10 |
| CN106367358A (en) * | 2016-09-29 | 2017-02-01 | 广东海洋大学 | Fungal aspergillus terreus strain C21-10 derived from coral |
| CN112920955B (en) * | 2021-02-09 | 2022-06-07 | 中国药科大学 | Deep-sea aspergillus terreus B12 capable of producing secondary metabolite with antibacterial and synergistic effects and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1629148A (en) * | 2004-10-10 | 2005-06-22 | 中国海洋大学 | Alkoxy ethoxy propyl isothiazolinone and its preparation process and use |
-
2014
- 2014-09-20 CN CN201410512799.2A patent/CN104293846B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1629148A (en) * | 2004-10-10 | 2005-06-22 | 中国海洋大学 | Alkoxy ethoxy propyl isothiazolinone and its preparation process and use |
Non-Patent Citations (2)
| Title |
|---|
| Antimicrobial butyrolactone I derivatives from the Ecuadorian soil fungus Aspergillus terreus Thorn. var terreus;M.E. Cazar et al.;《World Journal of Microbiology&Biotechnology》;20051031;第21卷;第1068页左栏倒数第1-2段和右栏第1段以及倒数第1段，第1069页图1和左栏第1段，第1073页表3 * |
| 海洋污损细菌群落结构及其发酵液中化学成分的研究;于洋等;《海洋科学》;20140915;第38卷(第9期);27-32 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104293846A (en) | 2015-01-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103554090B (en) | A kind of terpenoid dihydro-quinolinone alkaloid compound and its crystal, preparation method and the application as marine antifoulant | |
| CN101974464B (en) | Streptomyces and process for preparing antimycin antibiotics by fermentation using same | |
| CN107841466B (en) | A method of by marine Penicillum category fungi large scale preparation penicillic acid | |
| CN104293846B (en) | A kind of preparation method of butyrolactone compound and the application as marine antifoulant | |
| CN104031954B (en) | A kind of preparation method of monoterpene dihydro-quinolinone alkaloid compound and its crystal and the application as marine antifoulant | |
| CN110527629B (en) | Marine fungus-derived brefeldin A, preparation method and application thereof in resisting agricultural pathogenic bacteria | |
| CN105153103B (en) | A kind of preparation method of chloro polyketide and the application as marine antifoulant | |
| CN104230873B (en) | Isocoumarin compound as well as preparation method and application of isocoumarin compound as natural marine organism antifouling agent | |
| CN104031955B (en) | A kind of preparation method of dihydro-quinolinone alkaloid compound and the application as marine antifoulant | |
| CN105586373B (en) | Sclerotiorin derivative and preparation method thereof with as the application of marine antifoulant | |
| CN102105542A (en) | Antifouling furan-2-one derivatives | |
| CN103408490B (en) | A kind of preparation method and the application as environmentally friendly marine organisms stain control agent of quinolinone alkaloid | |
| CN103214364B (en) | A kind of preparation method and the application as natural marine organism stain control agent of Biphenyl Ether compound | |
| CN103554018B (en) | A kind of dihydro-quinolinone alkaloid compound and its crystal, preparation method and the application as marine antifoulant | |
| CN110003153B (en) | Benzofuran compound and preparation method and application thereof | |
| CN106496115A (en) | A kind of mixed source monoterpene alkaloid class compound and preparation method thereof and the application as marine antifoulant | |
| CN103351369B (en) | A kind of lignan compound and preparation method thereof and the application as natural marine organism stain control agent | |
| CN103882073B (en) | A kind of preparation method of macrolides compound and the application as marine antifoulant | |
| CN104163805B (en) | A kind of preparation method of mixed source chloro benzophenone and the application as marine antifoulant | |
| CN108794502B (en) | Trichothecene compound and preparation method and application thereof | |
| CN104630119B (en) | Algistatic activity material violacein and preparation method thereof | |
| CN104163757B (en) | A kind of mixed source Chloro benzophenone compound and preparation method thereof and the application as marine antifoulant | |
| CN108727169A (en) | A kind of preparation method of the biphenyl ether compound in marine fungi source and the application as antiseptic | |
| CN107973803B (en) | Seven-membered lactonofuran derivative and preparation method and application thereof | |
| CN102321089A (en) | Screw ring alkaloid compound, preparation method thereof and application in the aspect of marine organism fouling resistance |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |