CN104230873A - Isocoumarin compound as well as preparation method and application of isocoumarin compound as natural marine organism antifouling agent - Google Patents
Isocoumarin compound as well as preparation method and application of isocoumarin compound as natural marine organism antifouling agent Download PDFInfo
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- CN104230873A CN104230873A CN201310429588.8A CN201310429588A CN104230873A CN 104230873 A CN104230873 A CN 104230873A CN 201310429588 A CN201310429588 A CN 201310429588A CN 104230873 A CN104230873 A CN 104230873A
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- 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
- C07D311/76—Benzo[c]pyrans
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- 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
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
Abstract
The invention discloses an isocoumarin compound as well as a preparation method and application of the isocoumarin compound as a natural marine organism antifouling agent. The isocoumarin compound is characterized in that the compound has a structure as shown in formula I, wherein R2 and R2 independently represent H and a structure as shown in specification, and R1 or R2 is H; and R3 and R4 independently represent H and OCH3, and R3 or R4 is H. The compound as shown in formula I can be prepared through adopting such means as cultivation, fermentation and chromatographic separation on marine fungi, i.e., Eurotium sp. (HM991283). The natural marine organism antifouling agent disclosed by the invention is efficient and low in toxicity, and is characterized in that the compound of formula I or salt thereof, as an active ingredient, is used for preventing and controlling marine organism fouling caused by barnacle.
Description
Technical field
The present invention relates to a kind of Isocoumarin compounds, particularly relate to a kind of preparation method and the application as natural marine organism stain control agent with the Isocoumarin compounds of suppression kentrogon attachment activity.
Background technology
Marine biofouling refers to that marine fouling organism such as the larva of barnacle, mussel, polyzoan etc. forms group at the maritime facilitieies such as hull, fish box, oil well or the growth of other marine organisms surface attachment, thus cause huge infringement to being attached thing, as accelerated metallic corrosion, reducing maritime facilities performance, affecting culture fishery seed output and quality etc.In marine fouling organism, barnacle be distribute the widest, endanger a maximum class.The financial loss that the whole world causes due to marine biofouling is every year huge.For boats and ships, fouling organism adheres to alow, makes marine fuel consumption increase by 40%, and navigation cost increases by 77%, and global shipping loses about 3,000,000,000 dollars every year for this reason.The anti-fouling material of current use mainly contains sterilant or organometallic organic coating in facility surface coated such as hulls.These anti-fouling materials also havoc ocean environment while the anti-fouling effect of performance, as organotin oxides (TBTO) makes many fish and halobiontic immunity system be destroyed, and sterilant all produces murder by poisoning to target fouling organism and nontarget organism, and directly harm humans is healthy.Therefore, searching natural marine organism stain control agent that is efficient, low toxicity has become the key subjects that various countries face.The special conditionss such as ocean height salt, high pressure, low temperature, oligotrophic, low light photograph enable marine microorganism produce a large amount of novel structure, active unique secondary metabolite, wherein some compound has In Studies On Antifouling Activity, provides important sources for finding potential natural marine organism stain control agent.
Summary of the invention
The object of the present invention is to provide a kind of Isocoumarin compounds deriving from thalassiomycetes and preparation method thereof and the application as natural marine organism stain control agent, it can meet the demand of prior art.Culture presevation information: depositary institution's title: China Committee for Culture Collection of Microorganisms's common micro-organisms center; Depositary institution address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica;
Preservation date: on August 12nd, 2013; Deposit number: CGMCC7991; Classification And Nomenclature: Eurotium sp..
A kind of Isocoumarin compounds of formula I structure or its salt:
Wherein R
1, R
2be independently of one another H,
and R
1, R
2in have one for H; R
3, R
4be H, OCH independently of one another
3, and R
3, R
4in have one for H.
Formula I is selected from following compounds:
The invention provides a kind of preparation method of above-mentioned formula I, it is characterized in that comprising the following steps: first in bacterium culture medium, carry out spawn culture to thalassiomycetes Eurotium sp. (HM991283), then carry out fermentation culture to above-mentioned thalassiomycetes in the fermentation medium; After being separated with thalline by fermented liquid, fermented liquid is extracted with ethyl acetate, and extraction liquid concentrating under reduced pressure obtains fermented liquid medicinal extract; Thalline methyl alcohol lixiviate, concentrating under reduced pressure obtains thalline medicinal extract; Merge fermented liquid medicinal extract and thalline medicinal extract, carry out chromatographic separation, obtain formula I, containing glucose, yeast extract paste, peptone, agar, thick sea salt, water in wherein said bacterium culture medium, containing glucose, yeast extract paste, peptone, thick sea salt, water in fermention medium; Described chromatographic separation is that normal phase silica gel column chromatography separation is separated with high performance liquid phase chiral chromatography.
In above-mentioned preparation method, bacterium culture medium is preferably containing glucose 0.1%-5.0% (weight percent, lower same), yeast extract paste 0.01%-2%, peptone 0.01%-2%, agar 0.1%-3.0%, thick sea salt 0.05%-5%, and all the other are water; Culture temperature is preferably 5-45 DEG C; Incubation time is preferably 3-10 days; Fermention medium is preferably containing glucose 0.1%-5.0% (weight percent, lower same), yeast extract paste 0.01%-2%, peptone 0.01%-2%, thick sea salt 0.05%-5%, and all the other are water; Culture temperature is preferably 5-45 DEG C; Incubation time is preferably 5-50 days; The stationary phase adopted during described normal phase silica gel column chromatography is separated is preferably 200 ~ 300 order silica gel, and moving phase is preferably 10%-40% (volume percent, lower same) ethyl acetate-light petrol mixed solvent; The chromatographic column adopted during high performance liquid phase chiral chromatography is separated is preferably TBB chiral chromatographic column (Kromasil, 5 μm, 150 × 4.6mm), and moving phase is preferably 5%-15% Virahol-normal hexane mixed solvent.
The invention provides a kind of marine organisms stain control agent, it is characterized in that using above-mentioned formula I Isocoumarin compounds or its salt as effective constituent, for preventing and treating the marine biofouling that barnacle causes.
Isocoumarin compounds of the present invention has high inhibition effect to kentrogon attachment, and less to its toxicity, can be used for the marine organisms stain control agent of development environment close friend, and raw material can carry out scale operation by fungi fermentation, not by resource limit, therefore have a extensive future.
Embodiment
For the ease of a further understanding of the present invention, the embodiment provided below has done more detailed description to it.But these embodiments only are not used for limiting scope of the present invention or implementation principle for better understanding invention, embodiments of the present invention are not limited to following content.
Embodiment 1
(1) cultivation of thalassiomycetes Eurotium sp. (HM991283) bacterial classification
The substratum that the spawn culture of fungi Eurotium sp. (HM991283) is used contains glucose 1.0% (weight percent, down together), yeast extract paste 0.1%, peptone 0.2%, agar 1.0%, thick sea salt 3%, all the other are water, make test tube slant during use, fungal bacterial strain is cultivated 5 days at 28 DEG C.
(2) fermentation of thalassiomycetes Eurotium sp. (HM991283)
The fermention medium that the fermentation culture of fungi Eurotium sp. (HM991283) is used is containing glucose 1.0% (weight percent, down together), yeast extract paste 0.1%, peptone 0.2%, thick sea salt 3.0%, all the other are water, and fungal bacterial strain is large in 25 DEG C of cultivations 30.
(3) separation and Extraction of formula I
Get the fermented product 100L of step (2) gained, after being separated with thalline by fermented liquid, fermented liquid is extracted with ethyl acetate 5 times, and extraction liquid concentrating under reduced pressure obtains fermented liquid medicinal extract, thalline methyl alcohol lixiviate 5 times, concentrating under reduced pressure obtains thalline medicinal extract, merge fermented liquid medicinal extract and thalline medicinal extract, first carry out normal phase silica gel column chromatography separation, stationary phase: 200 ~ 300 order silica gel, moving phase: 10%-40% (volume percent, ethyl acetate-light petrol mixed solvent down together), separation obtains 6 raceme mixtures, then the separation of high performance liquid phase chiral chromatography is carried out respectively, stationary phase: TBB chiral chromatographic column (Kromasil, 5 μm, 150 × 4.6mm), moving phase: 5%-15% Virahol-normal hexane mixed solvent, separation obtains 12 pale yellow oil (compound 1-12), be formula I.
The structural identification data of gained formula I:
Compound 1,2
pale yellow oil; [α]
25 d1:+59; 2:-59; CD1: λ
max(Δ ε) 260 (-18.3), 340 (+2.8) nm; 2: λ
max(Δ ε) 260 (+18.3), 340 (-2.8) nm; 1,2 nuclear magnetic datas:
1h NMR (CDCl
3, 600MHz) and δ: 10.85 (1H, s, 2-OH), 6.94 (1H, s, H-4), 6.10 (1H, brs, 5-OH), 5.27 (1H, m, H-2 "), 4.69 (1H; d, J=2.4Hz, H-1 '), 4.43 (1H; ddd, J=8.4,5.4,2.4Hz; H-2 '), 3.35 (3H, s, 1 '-OCH
3), 3.32 (2H, d, J=7.2Hz, H-1 "), 1.96 (1H; m, Ha-3 '), 1.86 (1H, m, Hb-3 '); 1.74 (3H, s, H-4 "), 1.68 (3H, s, H-5 "), 1.55 (1H, m, Ha-4 '), 1.43 (1H; m, Hb-4 '), 1.32 (4H, m, H-5 '; H-6 '), 0.89 (3H, t, J=6.6Hz, H-7 ').
13c NMR (CDCl
3, 150MHz) and δ: 169.1 (C, C-7), 153.6 (C, C-2), 145.5 (C, C-5), 133.4 (C, C-3 "), 131.3 (C, C-3), 123.8 (CH; C-4), 120.1 (CH, C-2 "), 118.4 (C, C-6), 106.0 (C, C-1), 81.2 (CH, C-2 '), 68.3 (CH, C-1 '), 55.9 (CH
3, 1 '-OCH
3), 30.8 (CH
2, C-5 '), 29.3 (CH
2, C-3 '), 27.0 (CH
2, C-1 "), 25.0 (CH
3, C-4 "), 24.1 (CH
2, C-4 '), 21.7 (CH
2, C-6 '), 17.1 (CH
3, C-5 "), 13.2 (CH
3, C-7 ') and .HRESIMS m/z347.1860 [M-H]
-(calcd for C
20h
27o
5, 347.1853).
Compound 3,4
pale yellow oil; [α]
25 d3:+38; 4:-38; CD3: λ
max(Δ ε) 260 (-15.2), 340 (+2.0) nm; 4: λ
max(Δ ε) 260 (+15.2), 340 (-2.0) nm; 3,4 nuclear magnetic datas:
1h NMR (CDCl
3, 600MHz) and δ: 10.89 (1H, s, 2-OH), 6.93 (1H, s, H-4), 5.83 (1H, brs, 5-OH), 5.49 (1H, m, H-6 '), 5.42 (1H, m, H-5 '), 5.28 (1H, m, H-2 "), 4.71 (1H, d, J=2.4Hz; H-1 '), 4.47 (1H, ddd, J=7.8,5.4; 2.4Hz, H-2 '), 3.36 (3H, s, 1 '-OCH
3), 3.32 (2H, d, J=7.2Hz, H-1 "), 2.17-2.25 (2H, m, H-4 '); 2.06 (1H, m, Ha-3 '), 1.86 (1H; m, Hb-3 '), 1.75 (3H, s; H-4 "), 1.69 (3H, s, H-5 "); 1.64 (3H, t, J=6.0Hz, H-7 ').
13c NMR (CDCl
3, 150MHz) and δ: 169.6 (C, C-7), 154.4 (C, C-2), 145.2 (C, C-5), 134.1 (C, C-3 "); 132.1 (C, C-3), 129.6 (CH; C-5 '), 126.6 (CH, C-6 '); 124.5 (CH, C-4), 120.9 (CH; C-2 "), 119.0 (C, C-6), 106.9 (C, C-1), 80.8 (CH, C-2 '), 69.4 (CH, C-1 '), 56.6 (CH
3, 1 '-OCH
3), 29.8 (CH
2, C-3 '), 27.8 (CH
2, C-1 "), 25.8 (CH
3, C-4 "), 27.9 (CH
2, C-4 '), 18.0 (CH
3, C-7 '), 17.8 (CH
3, C-5 ") .HRESIMSm/z345.1706 [M-H]
-(calcd for C
20h
25o
5, 345.1697).
Compound 5,6
pale yellow oil; [α]
25 d5:+13; 6:-13; HRESIMS m/z345.1698 [M-H]
-(calcd for C
20h
25o
5, 345.1697).
Compound 7,8
pale yellow oil; [α]
25 d7:-58; 8:+58; CD7: λ
max(Δ ε) 260 (+18.2), 340 (-2.2) nm; 8: λ
max(Δ ε) 260 (-18.2), 340 (+2.2) nm; 7,8 nuclear magnetic datas:
1h NMR (CDCl
3, 600MHz) and δ: 10.95 (1H, s, 2-OH), 6.93 (1H, s, H-4), 5.82 (1H, brs, 5-OH), 5.28 (1H, m, H-2 "), 4.76 (1H; ddd, J=7.8,4.8,3.6Hz; H-2 '), 4.65 (1H, d, J=3.6Hz; H-1 '), 3.39 (3H, s, 1 '-OCH
3), 3.33 (2H, d, J=7.2Hz, H-1 "), 1.75 (3H; s, H-4 "), 1.69 (3H, s, H-5 "), 1.59 (1H; m, Ha-3 '), 1.54 (1H, m, Hb-3 '), 1.53 (1H; m, Ha-4 '), 1.41 (1H, m, Hb-4 '); 1.28 (2H, m, H-6 '), 1.27 (2H, m; H-5 '), 0.87 (3H, t, J=6.6Hz, H-7 ').
13c NMR (CDCl
3, 150MHz) and δ: 167.8 (C, C-7), 153.6 (C, C-2), 145.1 (C, C-5), 133.4 (C, C-3 "), 131.5 (C, C-3), 123.8 (CH; C-4), 120.0 (CH, C-2 "), 116.7 (C, C-6), 105.8 (C, C-1), 81.4 (CH, C-2 '), 70.4 (CH, C-1 '), 55.3 (CH
3, 1 '-OCH
3), 31.2 (CH
2, C-5 '), 30.6 (CH
2, C-3 '), 26.9 (CH
2, C-1 "), 25.0 (CH
3, C-4 "), 24.2 (CH
2, C-4 '), 21.6 (CH
2, C-6 '), 17.0 (CH
3, C-5 "), 13.1 (CH
3, C-7 ') and .HRESIMS m/z347.1860 [M-H]
-(calcd for C
20h
27o
5, 347.1853).
Compound 9,10
pale yellow oil; [α]
25 d9:-29; 10:+29; CD9: λ
max230 (+5.8), 258 (+10.5), 340 (-1.5) nm; (Δ ε) 10: λ
max(Δ ε) 230 (-5.8), 258 (-10.5), 340 (+1.5) nm; 9,10 nuclear magnetic datas:
1h NMR (CDCl
3, 600MHz) and δ: 10.96 (1H, s, 2-OH), 6.93 (1H, s, H-4), 5.63 (1H, brs, 5-OH), 5.50 (1H, m, H-6 '), 5.38 (1H, m, H-5 '), 5.28 (1H, m, H-2 "), 4.77 (1H, ddd, J=8.4; 4.8,3.6Hz, H-2 '), 4.65 (1H, d; J=3.6Hz, H-1 '), 3.39 (3H, s, 1 '-OCH
3), 3.33 (2H, d, J=7.2Hz, H-1 "), 2.18 (2H, m, H-4 '); 1.76 (3H, s, H-4 "), 1.70 (3H, s, H-5 "), 1.68 (2H, m; H-3 '), 1.64 (3H, dd; J=6.6,1.2Hz, H-7 ').
13c NMR (CDCl
3, 150MHz) and δ: 167.8 (C, C-7), 153.9 (C, C-2), 145.0 (C, C-5), 133.5 (C, C-3 "); 131.7 (C, C-3), 128.4 (CH; C-5 '), 126.2 (CH, C-6 '); 124.0 (CH, C-4), 120.1 (CH; C-2 "), 116.8 (C, C-6), 105.9 (C, C-1), 80.4 (CH, C-2 '), 70.9 (CH, C-1 '), 55.4 (CH
3, 1 '-OCH
3), 31.2 (CH
2, C-3 '), 27.4 (CH
2, C-4 '), 27.0 (CH
2, C-1 "), 25.2 (CH
3, C-4 "), 17.2 (CH
3, C-5 "), 17.1 (CH
3, C-7 ') and .HRESIMS m/z345.1706 [M-H]
-(calcd for C
20h
25o
5, 345.1697).
Compound 11,12
pale yellow oil; [α]
25 d11:-18; 12:+18; CD11: λ
max(Δ ε) 232 (+10.6), 255 (+7.5), 340 (-1.8) nm; 12: λ
max(Δ ε) 232 (-10.6), 255 (-7.5), 340 (+1.8) nm; 11,12 nuclear magnetic datas:
1h NMR (CD Cl
3, 600MHz) and δ: 10.91 (1H, s, 2-OH), 6.93 (1H, s, H-4), 6.28 (1H, brs, 5-OH), 5.94 (1H, m, H-4 '), 5.74 (1H, m, H-3 '), 5.29 (1H, m, H-2 "), 4.87 (1H, d; J=3.6Hz, H-1 '), 4.47 (1H, dd, J=8.4; 3.6Hz, H-2 '), 3.44 (3H, s, 1 '-OCH
3), 3.32 (2H, d, J=7.2Hz, H-1 "), 2.08 (2H, m; H-5 '), 1.76 (3H, s, H-4 "), 1.69 (3H, s, H-5 "); 1.42 (2H, m, H-6 '), 0.89 (3H, t, J=7.8Hz, H-7 ').
13c NMR (CDCl
3, 150MHz) and δ: 168.3 (C, C-7), 153.6 (C, C-2), 145.0 (C, C-5), 137.8 (CH, C-4 '), 133.4 (C, C-3 "), 131.4 (C; C-3), 124.2 (CH, C-4); 122.3 (CH, C-3 '), 120.1 (CH; C-2 "), 116.9 (C, C-6), 105.4 (C, C-1), 80.2 (CH, C-2 '), 71.6 (CH, C-1 '), 56.1 (CH
3, 1 '-OCH
3), 33.6 (CH
2, C-5 '), 26.9 (CH
2, C-1 "), 25.1 (CH
3, C-4 "), 21.2 (CH
2, C-6 '), 17.0 (CH
3, C-5 "), 12.8 (CH
3, C-7 ') and .HRESIMSm/z345.1698 [M-H]
-(calcd for C
20h
25o
5, 345.1697).
Embodiment 2
(1) cultivation of thalassiomycetes Eurotium sp. (HM991283) bacterial classification
Bacterium culture medium contains glucose 0.1%-5.0% (weight percent, lower same), yeast extract paste 0.01%-2%, peptone 0.01%-2%, agar 0.1%-3.0%, thick sea salt 0.05%-5%, and all the other are water; Culture temperature is 5-45 DEG C; Incubation time is 3-10 days.
(2) fermentation of thalassiomycetes Eurotium sp. (HM991283)
Fermention medium contains glucose 0.1%-5.0% (weight percent, lower same), yeast extract paste 0.01%-2%, peptone 0.01%-2%, thick sea salt 0.05%-5%, and all the other are water; Culture temperature is 5-45 DEG C; Incubation time is 5-50 days.
(3) separation and Extraction of formula I
Get the fermented product 50L of step (2) gained, after being separated with thalline by fermented liquid, fermented liquid is extracted with ethyl acetate 2 ~ 6 times, and extraction liquid concentrating under reduced pressure obtains fermented liquid medicinal extract, thalline methyl alcohol lixiviate 2 ~ 6 times, concentrating under reduced pressure obtains thalline medicinal extract, merge fermented liquid medicinal extract and thalline medicinal extract, first carry out normal phase silica gel column chromatography separation, stationary phase: 200 ~ 300 order silica gel, moving phase: 10%-40% (volume percent, ethyl acetate-light petrol mixed solvent down together), separation obtains 6 raceme mixtures, then the separation of high performance liquid phase chiral chromatography is carried out respectively, stationary phase: TBB chiral chromatographic column (Kromasil, 5 μm, 150 × 4.6mm), moving phase: 5%-15% Virahol-normal hexane mixed solvent, separation obtains 12 pale yellow oil (compound 1-12), be formula I.The structural identification data of its compounds of formula I are consistent with corresponding data in embodiment 1.
Other spawn culture, the fermentation condition that specifically do not indicate in embodiment 1-2, and other experimental operating conditions such as normal phase silica gel column chromatography separation, the separation of high performance liquid phase chiral chromatography are the experimental operating conditions of this area routine, those skilled in the art can according to actual needs, reasonably select.
Embodiment 3
In bacterium culture medium, spawn culture is carried out to thalassiomycetes Eurotium sp. (HM991283), in the fermentation medium fermentation culture is carried out to above-mentioned thalassiomycetes again, after fermented liquid is separated with thalline, fermented liquid is extracted with ethyl acetate, and extraction liquid concentrating under reduced pressure obtains fermented liquid medicinal extract; Thalline methyl alcohol lixiviate, concentrating under reduced pressure obtains thalline medicinal extract; Merge fermented liquid medicinal extract and thalline medicinal extract, obtain 12 pale yellow oil (compound 1-12) after carrying out chromatographic separation and be formula I, its structural identification data are consistent with corresponding data in embodiment 1.Containing glucose, yeast extract paste, peptone, agar, thick sea salt, water in wherein said bacterium culture medium, containing glucose, yeast extract paste, peptone, thick sea salt, water in described fermention medium; Described chromatographic separation is that normal phase silica gel column chromatography separation is separated with high performance liquid phase chiral chromatography.
In order to explore the method being applicable to widely prepare formula I, the interpolation of each composition in bacterium culture medium, fermention medium in the present embodiment, all add in ratio conventional in this area or add in any proportion, the selection of the specification of used silica gel, the model of chiral chromatographic column and eluting solvent during chromatographic separation, the routine being this area is selected.Experimental result shows, the preparation method that above-mentioned routine is selected, all can obtain 12 pale yellow oil invented, i.e. formula I, its structural identification data are consistent with corresponding data in embodiment 1, only there is the fine difference of individual compound in purity and yield.
The result of embodiment 1-3 shows, according to spawn culture, the fermentation condition of this area routine, the condition that conventional normal phase silica gel column chromatography is separated, high performance liquid phase chiral chromatography is separated carries out spawn culture, fermentation, separation and purification to thalassiomycetes Eurotium sp. (HM991283), all can obtain the compound of formula I structure.The preparation method of formula I, the method recorded in preferred embodiment 1-2.
Embodiment 4
The anti-kentrogon attachment activity (table 1) of formula I.
Formula I of the present invention is tested according to following literature method the test of barnacle Balanus amphitrite larva attachment inhibit activities: Thiyagarajan V.; Harder T.; Qiu J.W.; Qian P.Y.Mar Biol (Berl) 2003,143,543-554.By suppressing the test of barnacle B.amphitrite larva attachment activity, find the medium effective concentration (EC of formula I of the present invention (i.e. compound 1-12)
50) be all less than 25 μ g/mL, its median lethal concentration (LC
50) be all greater than 50 μ g/mL.This shows that formula I is in the attachment of suppression barnacle B.amphitrite larva, all shows the feature of high-efficiency low-toxicity.Wherein, compound 8 inhibit activities is the strongest, medium effective concentration (EC
50) be 0.73 μ g/mL (table 1).Only list in table 1 the compounds of this invention 1-3,5,7,8,10, the Activity Results of 12.
Table 1 segment bounds I of the present invention adheres to inhibit activities and toxicity to kentrogon B.amphitrite
Isocoumarin class formula I of the present invention has strong suppression barnacle attachment activity, and it is lower to the toxicity of kentrogon, natural marine organism stain control agent that is efficient, low toxicity can be made into, and raw material can carry out scale operation by fungi fermentation, not by resource limit, therefore have a extensive future.
Claims (8)
1. the Isocoumarin compounds of a formula I structure or its salt:
Wherein R
1, R
2be independently of one another H,
and R
1, R
2in have one for H; R
3, R
4be H, OCH independently of one another
3, and R
3, R
4in have one for H.
2. formula I as claimed in claim 1, is characterized in that being selected from following compound:
3. the preparation method of the formula I described in claim 1 or 2, it is characterized in that comprising the following steps: first in bacterium culture medium, carry out spawn culture to thalassiomycetes Eurotium sp. (HM991283), in the fermentation medium fermentation culture is carried out to above-mentioned thalassiomycetes again, after fermented liquid is separated with thalline, fermented liquid is extracted with ethyl acetate, and extraction liquid concentrating under reduced pressure obtains fermented liquid medicinal extract; Thalline methyl alcohol lixiviate, concentrating under reduced pressure obtains thalline medicinal extract; Merge fermented liquid medicinal extract and thalline medicinal extract, carry out chromatographic separation, obtain formula I; Containing glucose, yeast extract paste, peptone, agar, thick sea salt, water in wherein said bacterium culture medium; Containing glucose, yeast extract paste, peptone, thick sea salt, water in described fermention medium; Described chromatographic separation is that normal phase silica gel column chromatography separation is separated with high performance liquid phase chiral chromatography.
4. preparation method according to claim 3, it is characterized in that described bacterium culture medium contains glucose 0.1%-5.0%, yeast extract paste 0.01%-2%, peptone 0.01%-2%, agar 0.1%-3.0%, thick sea salt 0.05%-5%, all the other are water, and above-mentioned percentage composition is all weight percentage; Culture temperature is 5-45 DEG C; Incubation time is 3-10 days.
5. preparation method according to claim 3, it is characterized in that described fermention medium contains glucose 0.1%-5.0%, yeast extract paste 0.01%-2%, peptone 0.01%-2%, thick sea salt 0.05%-5%, all the other are water, and above-mentioned percentage composition is all weight percentage; Culture temperature is 5-45 DEG C; Incubation time is 5-50 days.
6. preparation method according to claim 3, it is characterized in that the stationary phase adopted during described normal phase silica gel column chromatography is separated is 200 ~ 300 order silica gel, moving phase is the ethyl acetate-light petrol mixed solvent of 10%-40%; The chromatographic column adopted during described high performance liquid phase chiral chromatography is separated is TBB chiral chromatographic column Kromasil, 5 μm, 150 × 4.6mm, and moving phase is the Virahol-normal hexane mixed solvent of 5%-15%; Above-mentioned mixed solvent per-cent is volume percent.
7. a marine organisms stain control agent, is characterized in that containing the compound or its salt described in claim 1 or 2 as effective constituent.
8. the application in the marine biofouling that the formula I described in claim 1 or 2 or its salt cause at control barnacle.
Priority Applications (1)
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| CN109456292A (en) * | 2018-10-23 | 2019-03-12 | 中山大学 | A kind of coumarin kind compound and the preparation method and application thereof in marine fungi source |
| CN110078720A (en) * | 2019-05-28 | 2019-08-02 | 扬州工业职业技术学院 | Halogenated benzofuran-coumarin derivative and its preparing the application in fouling resistance agent |
| CN110105345A (en) * | 2019-05-28 | 2019-08-09 | 扬州工业职业技术学院 | The cumarin amine derivative and its application in fouling resistance that benzofuran replaces |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104327035A (en) * | 2013-12-14 | 2015-02-04 | 于跃 | Marine fungi secondary metabolite derivative and application of marine fungi secondary metabolite derivative as marine biological antifoulant |
| CN104327035B (en) * | 2013-12-14 | 2017-03-22 | 于跃 | Marine fungi secondary metabolite derivative and application of marine fungi secondary metabolite derivative as marine biological antifoulant |
| CN104710396B (en) * | 2013-12-17 | 2017-04-12 | 扬州大学 | Ocean-originated fungus secondary metabolite derivatives and application of same as anti-MRSA drug |
| CN109456292A (en) * | 2018-10-23 | 2019-03-12 | 中山大学 | A kind of coumarin kind compound and the preparation method and application thereof in marine fungi source |
| CN109456292B (en) * | 2018-10-23 | 2022-06-10 | 中山大学 | Coumarin compound derived from marine fungi as well as preparation method and application of coumarin compound |
| CN110078720A (en) * | 2019-05-28 | 2019-08-02 | 扬州工业职业技术学院 | Halogenated benzofuran-coumarin derivative and its preparing the application in fouling resistance agent |
| CN110105345A (en) * | 2019-05-28 | 2019-08-09 | 扬州工业职业技术学院 | The cumarin amine derivative and its application in fouling resistance that benzofuran replaces |
| CN110078720B (en) * | 2019-05-28 | 2020-07-17 | 扬州工业职业技术学院 | Halogenated benzofuran-coumarin derivative and application thereof in preparation of anti-fouling agent |
| CN110105345B (en) * | 2019-05-28 | 2020-07-17 | 扬州工业职业技术学院 | Benzofuran substituted coumarin amine derivative and application thereof in pollution resistance |
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