CN106632230A - Aspergillus unguis bromo-depsidone compound and preparation method and application thereof - Google Patents
Aspergillus unguis bromo-depsidone compound and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses an aspergillus unguis bromo-depsidone compound and a preparation method and application thereof. The chemical structure formula of the aspergillus unguis bromo-depsidone compound is shown as the formula (I) in the accompanying drawing. The compound is obtained by inoculating aspergillus unguis DLEP2008001 into a fungus culture medium; performing still standing fermentation; extracting fermentation liquid by ethyl acetate; extracting mycelia by an organic solvent; after the merging, performing gradient silicagel column chromatography by an organic solvent; through the silicagel thin layer chromatography detection, continuously performing silicagel column and gel column chromatography on the elution fractions containing target subjects; performing normal phase preparation liquid chromatographic separation purification. The compound can be used for inhibiting fungus white candida, gram-negative bacterium pseudomonas aeruginosa and gram-positive bacterium methoxyl-resistant penicillin staphylococcus aureus and killing artemia larvae, and the potential purposes of antifungal antibiotics, antibacterial antibiotics or pesticides are realized.
Description
Technical field
The invention belongs to microbial fermenters extractive technique field, in particular it relates to a kind of marine fungi pawl aspergillus bromo
Depsidone class compound and its preparation method and application.
Background technology
The drug resistance of the current microorganism that finds the cause of disease in clinical treatment is more and more stronger, especially the penicillin of resistance to methoxyl group gold
Staphylococcus aureus（MRSA）, pseudomonas aeruginosa（It is commonly called as Pseudomonas aeruginosa）And Candida albicans（It is commonly called as Candida albicans）Make
Into infection it is increasingly severe, many conventional antibiotics have lost curative effect substantially, it is therefore necessary to find new antibiotic come
Tackle this challenge.Additionally, agricultural production because excessively use various chemical synthetic pesticides, cause serious food, soil and
Water pollution, and jeopardize health and the ecological balance through food chain, residues of pesticides also seriously hinder the agricultural product of China and go out
Mouthful；And biological pesticide has the advantages that to be not likely to produce the resistance to the action of a drug, to non-target organism safety, environmental friendliness and is easy to natural degradation,
For human health, environmental protection and agricultural sustainable development have great importance.Antibiotic and kill that marine fungi is originated
Worm agent have the advantages that to be easy to large scale fermentation production, efficiently, be easy to overcome the resistance to the action of a drug, it is easy to accomplish medicine source stable supplying, tool
There are preferable economic, society and environmental benefit.
The content of the invention
The technical problem to be solved is the defect and deficiency for overcoming existing antibiotic and insecticide, there is provided a kind of
Extract from the bromo depsidone class compound of marine fungi pawl aspergillus, the compound have antimycotic, antibacterial activity and
Insecticidal activity, is with a wide range of applications in antifungal agent, antibacterial agent and agricultural insecticide is prepared.
It is an object of the invention to provide a kind of marine fungi pawl aspergillus bromo depsidone class compound.
It is a further object of the present invention to provide the preparation method of above-claimed cpd.
Another object of the present invention is to provide the application of above-claimed cpd.
The above-mentioned purpose of the present invention is that used technical scheme below gives realization.
The marine fungi pawl aspergillus that the present invention is adopted（Aspergillus unguis）CGMCC No.3372 bacterial classifications, in
On October 28th, 2009 is stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center（CGMCC）, preservation volume
Number be CGMCC No.3372, strain number be DLEP2008001.
A kind of marine fungi pawl aspergillus（Aspergillus unguis）Bromo depsidone class chemicals, its feature exists
In the chemical structural formula of the chemicals is such as（）It is shown：
（）
The preparation method of above-mentioned marine fungi pawl aspergillus bromo depsidone class chemicals comprises the steps：
S1. ferment：Pawl aspergillus DLEP2008001 is inoculated in fungi culture medium after standing for fermentation, respectively collect mycelium and
Zymotic fluid；；
S2. slightly carry：The zymotic fluid of step S1 is concentrated Jing after ethyl acetate extraction, mycelium is concentrated Jing after Solvent Extract methods,
Crude extract is obtained after merging；
S3. isolate and purify：The crude extract of step S2 is carried out into silicagel column gradient chromatography, the detection of Jing silica gel thin-layer chromatographies will contain
The elution fraction of target substance proceeds silicagel column, gel filtration chromatography and positive preparative liquid chromatography and isolates and purifies and obtain target
Compound；
Wherein, the formula of fungi liquid culture medium described in step S1 is：Contain following component in per liter：Murphy juice 400～600
15～25g of mL, NaBr, 15～25g of sucrose, pure water or the mL of running water 400～600.
Preferably, the formula of the fungi liquid culture medium is：Contain following component in per liter：The mL of murphy juice 500,
The g of NaBr 20, the g of sucrose 20, pure water or the mL of running water 500.
Specifically, the preparation method of above-mentioned marine fungi pawl aspergillus bromo depsidone class chemicals includes following step
Suddenly：
S1. ferment：By pawl aspergillus（Aspergillus unguis）CGMCC No.3372 are inoculated in fungi liquid culture medium
Standing for fermentation, after fermenting 2～4 days, adds final concentration of 0.5～2 mM（It is preferred that 1mM）Anuject, continue ferment 18
～22 days, filter, mycelium and zymotic fluid are collected respectively；
S2. slightly carry：The zymotic fluid Jing ethyl acetate that step S1 is obtained is extracted and concentrated, the mycelium that step S1 is obtained is used
Solvent Extract methods are simultaneously concentrated, and gained concentrate are merged, as crude extract；
Wherein, the organic solvent is one or more in chloroform, acetone, ethyl acetate, ethanol, methyl alcohol；
S3. isolate and purify：A. the crude extract Jing silica gel column chromatographies for step S2 being obtained, with petroleum ether-dichloromethane or oil
Ether-chloroform carries out pre- wash-out, carries out gradient elution with chloroform-methanol or methylene chloride-methanol then, collects with effluent volume
Than 100:0～100:The component of the wash-out of 1 gradient；Component characteristicses are that solvent is volume ratio 15 on silica gel thin-layer chromatography plate:1
Under chloroform-methanol launches, Rf value is 0.29～0.74 component of mixture；
B. the elution fraction that step a is collected is carried out into again silica gel column chromatography, eluent is volume ratio 5:1 petroleum ether-acetone；
C. the elution fraction that step b is collected is carried out into sephadex column chromatography, eluent is methyl alcohol, flow velocity 0.5～0.7
mL/min；
D. the elution fraction that step c is collected is carried out into positive preparative liquid chromatography purifying, eluent is volume ratio 3～5:1（It is excellent
Select 4:1）Chloroform-petroleum ether, silicagel column fill 20～30g（It is preferred that 25 g）Silica gel, the mL/min of flow velocity 2～4（It is preferred that 3
mL/min）In the case of, collect the component that retention time is 14～28 min；
E. the elution fraction that step d is collected is proceeded into positive preparative liquid chromatography purifying, eluent is pure chloroform, in silicon
Glue post fills 10～15g（It is preferred that 12 g）Silica gel, the mL/min of flow velocity 1～3（It is preferred that 2mL/min）In the case of, collect retention time
For the main peak of 14～35 min, gleanings are obtained.
Preferably, the silica gel of step a or silica gel column chromatography described in b is 200～300 mesh.
In addition, specifically, Rf value and chemical colour reaction of the bromo depsidone class compound on thin-layer chromatography
Feature is as follows：When chromatography solvent is pure chloroform, Rf value of the compound on GF254 thin layer silica gel plates is 0.26,254nm
It is in single, uniform black～skipper spot that ultraviolet irradiation is lower, and the colour developing of anisaldehyde sulfuric acid is sepia.
Preferably, compound retention time under the conditions of high-efficient liquid phase analysis is the single chromatogram of 4.0～5.0 min
Peak.
It is highly preferred that the efficient liquid phase chromatographic analysis condition is Acquity UPLC BEH C18 posts, packing material size
1.7 microns, mm × 50 mm of column dimension 2.1, mobile phase is the acetonitrile solution of the modifying agent formic acid for adding a ten thousandth volume,
0～2.5 min acetonitriles ratio is 60%, and 2.5～2.6 min acetonitrile proportional linearities rise to 100%, 2.6～3.0 min acetonitrile ratios
For 100%, 3.0～3.1 min acetonitrile proportional linearities are down to 60%, and 3.1～5.0 min acetonitriles ratios are 60%, the ml/ of flow velocity 0.3
min。
Preferably, the bromo depsidone class compound is at normal temperatures and pressures colorless needles.
Jing antibacterial experiments show that compound of the present invention is 6.4 μm of ol/L to the MIC value of Candida albicans, is had
Suppress the activity of Candida albicans；It is 1.6 μm of ol/L to the MIC of pseudomonas aeruginosa, shows that the compound has and suppress copper
The activity of green pseudomonad growth；It is 25.6 μm of ol/L to the MIC value of the staphylococcus aureus of the penicillin of resistance to methoxyl group, has
There is the activity for suppressing MRSA；It is 12.0 μm of ol/L to the half lethal dose LC50 of artemia, shows that it has insecticidal activity.
So, application of the above-mentioned bromo depsidone class chemicals on antimycotic, antibacterium or desinsection, and in system
Application in standby antifungal agent, antibacterial agent or insecticide, all should be within protection scope of the present invention.
Preferably, the fungi is Candida albicans.
Preferably, the bacterium is Gram-negative bacteria.
It is highly preferred that the Gram-negative bacteria is pseudomonas aeruginosa.
In addition, it is highly preferred that the bacterium is pseudomonas aeruginosa or the penicillin of resistance to methoxyl group staphylococcus aureus.
Preferably, the worm is agricultural pests.
It is highly preferred that the agricultural pests are the evil such as artemia, nematode, beet armyworm, aphid, Ability of Culex Pipiens Larvae or Tetranychus cinnabarinus
Worm.
The present invention compared with prior art, has the advantages that：
Present invention obtains a kind of new marine fungi pawl aspergillus bromo depsidone class compound, with good anti-true
Bacterium, antibacterial activity and insecticidal activity, and compounds process for production thereof is simple, it is easy to mass produce, prepare antifungal agent,
It is with a wide range of applications in antibacterial agent or agricultural insecticide.
Description of the drawings
Fig. 1 is the proton nmr spectra of bromo depsidone class compound of the present invention.
Fig. 2 is the carbon-13 nmr spectra of bromo depsidone class compound of the present invention.
Fig. 3 is the ESI of bromo depsidone class compound of the present invention-Mass spectrum.
Specific embodiment
The present invention is made with reference to Figure of description and specific embodiment further being elaborated, the embodiment
It is served only for explaining the present invention, is not intended to limit the scope of the present invention.Test method used in following embodiments is such as without spy
Different explanation, is conventional method；Material, reagent for being used etc., are the reagent for commercially obtaining if no special instructions
And material.
Marine fungi pawl aspergillus of the present invention（Aspergillus unguis）CGMCC No.3372 bacterial classifications, in 2009
On October 28, in is stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center（CGMCC）, deposit number is
CGMCC No.3372, strain number is DLEP2008001.
The preparation method of the marine fungi bromo depsidone class compound of embodiment 1
1. ferment
By pawl aspergillus（Aspergillus unguis）CGMCC No.3372 are inoculated in static fermentation in fungi liquid culture medium,
After fermentation 2 days, the Anuject of final concentration of 1mM is added, continue to ferment 20 days, filtered, mycelium is collected respectively and is sent out
Zymotic fluid.
Wherein, the formula of the fungi liquid culture medium is：
Contain following component in per liter：The mL of murphy juice 500, NaBr 20 g, the g of sucrose 20, pure water or the mL of running water 500.
2. slightly carry
The zymotic fluid Jing ethyl acetate that step 1 is obtained is extracted three times, and reduced pressure concentration obtains concentrate.Step 1 is obtained simultaneously
With there is methanol extract liquid to extract three times, reduced pressure concentration obtains concentrate to mycelium.Gained concentrate is merged, as crude extract；
3. isolate and purify
A. the crude extract Jing silica gel column chromatographies for step 2 being obtained, with petroleum ether-chloroform pre- wash-out is carried out, then with chloroform-first
Alcohol carries out gradient elution, and collection compares 100 with effluent volume:0～100:The component of the wash-out of 1 gradient；Component characteristicses are silica gel
Solvent is volume ratio 15 in thin layer chromatography board:Under 1 chloroform-methanol launches, Rf value is 0.29～0.74 mixing
Thing component；
B. the elution fraction that step a is collected is carried out into again silica gel column chromatography, eluent is volume ratio 5:1 petroleum ether-acetone；
C. the elution fraction that step b is collected is carried out into sephadex column chromatography, eluent is methyl alcohol, flow velocity 0.5～0.7
mL/min；
D. the elution fraction that step c is collected is carried out into positive preparative liquid chromatography purifying, eluent is volume ratio 4:1 chloroform-
Petroleum ether, in the case of silicagel column fills 25 g silica gel, the mL/min of flow velocity 3, collects the group that retention time is 14～28 min
Point；
E. the elution fraction that step d is collected is proceeded into positive preparative liquid chromatography purifying, eluent is pure chloroform, in silicon
Glue post is filled in the case of 12 g silica gel, the mL/min of flow velocity 2, collects the main peak that retention time is 14～35 min；Collected target
Material also is compliant with following characteristics：
（1）TLC detections are collected under the irradiation of 254nm ultraviolets in single, uniform black～skipper spot, anisaldehyde sulfuric acid
Develop the color for tan material；
（2）Rf values are 0.26 material under the conditions of the thin-layer chromatography of detection；Described detection thin layer chromatography board is GF254
Silica gel plate, solvent is pure chloroform；
（3）Retention time is the component of the single chromatographic peak of 4.36 min under the conditions of the ultra high efficiency liquid phase analysis of detection；Institute
The detection efficient liquid phase chromatographic analysis condition stated is Acquity UPLC BEH C18 posts, 1.7 microns of packing material size, post chi
Very little 2.1 mm × 50 mm, mobile phase be add a ten thousandth volume modifying agent formic acid acetonitrile solution, 0～2.5 min second
Nitrile ratio is 60%, and 2.5～2.6 min acetonitrile proportional linearities rise to 100%, and 2.6～3.0 min acetonitriles ratios are 100%, 3.0～
It is 60% that 3.1 min acetonitrile proportional linearities are down to 60%, 3.1-5.0 min acetonitriles ratio, the ml/min of flow velocity 0.3.
4th, the compound that the above-mentioned separation processes of Jing are obtained, is defined as pure compound.
Jing Spectrum Analysis, Structural Identification result shows that it is a kind of bromo depsidone class natural products, and structural formula is such as
（I）It is shown：
。
（I）
The compound has following physics and chemistry and spectral characteristic：
Colorless needles, proton nmr spectra (1H NMR, CDCl3, 500 MHz) δ6.76 (s), 5.38 (q, 6.8),
2.46 (s), 2.33 (s), 1.95 (s), 1.82 (dd, 6.8,1.0), as shown in Figure 1.
Carbon-13 nmr spectra (13C NMR, CDCl3, 125 MHz) δ162.0 (s), 160.2 (s), 156.5 (s),
148.8 (s), 145.3 (s), 142.9 (s), 142.5 (s), 136.1 (s), 132.1 (s), 128.1 (d),
116.1 (s), 115.3 (d), 114.5 (s), 108.0 (s), 99.6 (s), 21.7 (q), 17.9 (q),
14.5 (q), 10.6 (q), as shown in Figure 2.
ESI-Mass spectrum show quasi-molecular ion peak [M-H]-isotopic peak cluster mass-to-charge ratio be m/z 481/483/485, its
Peak intensity is 1：2：1, be typical two bromos feature and with molecular formula C19H17O5Br2It is consistent, as shown in Figure 3.
The fungicidal activities experiment of the bromo depsidone class compound of embodiment 2
1st, Candida albicans（Candida albicans）It is a kind of common opportunistic fungus, many portions of human body can be invaded
Position, causes cutaneous candidiasis, candidiasis of the mucous membranes（Thrush, angular stomatitis, vaginitis are most common）And internal organ and maincenter god
Jing candidiasis, is modal nosomycosis.Clinical symptoms are intricate, suddenly slow to differ.Recently as antibiotic, hormone,
The heavy dose application of immunodepressant, and the development of transplant operation, the candidiasis incidence of disease gradually increases, and can jeopardize life
Life causes serious consequence.The bacterium has become the important for target of antifungal drug research and development.
2nd, experimental technique：
The micro broth dilution method adopted international standards（National Committee for Clinical Laboratory
Standards. 1997. Reference methed for broth dilution antifungal
susceptibility testing of yeasts: approved standard M27-A, 7th ed. NCCLS,
Wayne, Pa.）Determine minimal inhibitory concentration of the compound to Candida albicans（MIC value）.
Control group is set simultaneously：Amphotericin B.
3rd, experimental result：
As a result show, the bromo depsidone class compound that above-described embodiment 1 is obtained is to the MIC value of Candida albicans
6.4 μm of ol/L, the activity with significant suppression Candida albicans.
And, amphotericin B is 12.8 μm of ol/L to the MIC value of Candida albicans, bromo depsidone of the present invention
Class compound is significantly stronger than positive control amphotericin B to the inhibitory activity of Candida albicans.
The suppression gram prolapse of uterus activity experiment of the bromo depsidone class compound of embodiment 3
1st, pseudomonas aeruginosa（Pseudomonas aeruginosa）It is a kind of conditioned pathogen also known as Pseudomonas aeruginosa, is also
One of the main pathogenic fungi of inside-hospital infection.The patient for suffering from metabolic disease, blood disease and malignant tumour, and it is postoperative or some
Patient susceptible after treatment contaminates this bacterium.The bacterium often causes postoperative wound infection, can also cause bedsore, abscess, otitis media suppurative
Deng.The infection focus that it causes can cause hematogenous extension, and bacteremia and septicemia occur.Infections after burn verdigris color is false single
Born of the same parents bacterium can cause death.And, with the extensive Clinical practice of broad-spectrum antibiotic, hormone and immunodepressant, verdigris is false single
Born of the same parents bacterium quickly generates drug resistance to Multiple Classes of Antibiotics so that clinical anti-infective therapy seems more and more difficult.At present, pin
It is also an emphasis of antibiotic research and development to the medicament research and development of the bacterium.
2nd, experimental technique：
The micro broth dilution method adopted international standards（National Committee for Clinical Laboratory
Standards. 2003. Methods for dilution antimicrobial susceptibility tests for
bacteria that grow aerobically, 5th ed. Approved standard M7–A6.）Determine the compound
Minimal inhibitory concentration to pseudomonas aeruginosa（MIC value）.
3rd, experimental result：
The bromo depsidone class compound that above-described embodiment 1 is obtained is 1.6 μm of ol/L to the MIC of pseudomonas aeruginosa,
Show that the compound has the activity for suppressing P. aeruginosa growth well.
The suppression Gram-positive drug-fast bacteria activity experiment of the bromo depsidone class compound of embodiment 4
1st, staphylococcus aureus（Staphylococcus aureus）It is very common germ, sometimes it can enter people
Cause infection in vivo.This infection slight meeting papula and papule on skin, serious can then cause pneumonia or blood sense
Dye.The infection caused to staphylococcus is generally treated with the antibiotic methicillin of PCs, in most cases very
Effectively.But some aureus strains define the resistance to the action of a drug, that is, the golden yellow of the penicillin of resistance to methoxyl group to methicillin
Staphylococcus（methicillin—resistantStaphylococcus aureus, MRSA）.Find in Britain within 1961
The first MRSA, worldwide spreads with surprising rapidity afterwards, is clinical modal Gram-positive drug-resistant bacteria,
According to estimates annual about 100,000 people are because infection MRSA and hospitalization, be also antibiotics research and development important target cause of disease it is micro-
It is biological.
2nd, experimental technique：
The micro broth dilution method adopted international standards（National Committee for Clinical Laboratory
Standards. 2003. Methods for dilution antimicrobial susceptibility tests for
bacteria that grow aerobically, 5th ed. Approved standard M7–A6.）Determine compound pair
The staphylococcus aureus of the penicillin of resistance to methoxyl group（MRSA）Minimal inhibitory concentration（MIC value）.
3rd, experimental result：
The MIC value of the bromo depsidone class compound that above-described embodiment 1 is obtained is 25.6 μm of ol/L, with good
Suppress the activity of MRSA.
The insecticidal activity experiment of the bromo depsidone class compound of embodiment 5
1st, artemia（brine shrimp）Also known as salt solution fairy shrimp, category Arthropoda, Crustachia, Anostraca, salt solution fairy shrimp
Section, genus artemia, are that a kind of fish food with higher economic worth is biological, are also a kind of important for toxin more sensitivity
Animal used as test, the activity that 2 ~ 3 instar larvaes of its laboratory cultures can be used to evaluate insecticide is strong and weak.
2nd, insecticidal activity experimental technique（Applicant paper Toxicology Mechanisms and Methods are referred to,
2012, 22(1), 23~30）.
3rd, artemia hatching and collection：A dual chamber linker is added through the frozen slag oxygenation of Indoor Natural light stimulus recovery
Cultivate in the hatchery of formula culture apparatus, the seawater that natural coarse sea salt is prepared is filled in linker（30 g/L）, ovum injected volume is 1
G/L, first closes linker passage.Jing after the hatching in 24 hours of about 2000 lux illumination at 28 DEG C, linker passage is opened, and will
Hatchery shading, and collecting pit keeps the h of illumination 12 ~ 24, carries out photoinduction；Linker passage is then shut off, with opening smoothness
The artemia larva that liquid transfer gun head is drawn in collecting pit carries out follow-up test.
4th, the biological lethal method of artemia：Sample is diluted into series concentration in 96 orifice plates with the continuous sesquialter of absolute methanol, very
Sky is dried and removes organic solvent, and 200 microlitres of artemia larva suspensions are added per hole（Containing 20 ~ 30 polypides）, and blank is set
Group, larva sum and death toll, correction of the calculating per hole in binocular stereo counted under microscope is per hole after 28 DEG C of 24 h of culture
The death rate,
Corrected mortality=（The control group death rate-disposal hole the death rate）/ control group survival rate × 100%；
Figure is done with the log2 logarithms of corrected mortality-concentration, in linear preferable 10% ~ 90% death rate interval linear trend is taken
Line, calculates half lethal dose LC50.
5th, experimental result：
The half lethal dose LC50 of the bromo depsidone class compound of above-mentioned acquisition is 12.0 μm of ol/L, shows its tool
There is insecticidal activity.
And, copper sulphate is 102.4 μm of ol/L to the LC50 of artemia, and bromo depsidone class compound is to artemia
Insecticidal activity is better than positive control copper sulphate.
In addition, research show, the compound to insects such as nematode, beet armyworm, aphid, Ability of Culex Pipiens Larvae or Tetranychus cinnabarinus,
There is preferable insecticidal activity.
Claims (10)
1. a kind of marine fungi pawl aspergillus bromo depsidone class compound, it is characterised in that the chemistry knot of the compound
Structure formula such as formula（）It is shown：
。
2. the preparation method of bromo depsidone class compound described in claim 1, it is characterised in that methods described include as
Lower step：
S1. ferment：Pawl aspergillus DLEP2008001 is inoculated in fungi liquid culture medium after standing for fermentation, mycelia is collected respectively
Body and zymotic fluid；
S2. slightly carry：The zymotic fluid of step S1 is concentrated Jing after ethyl acetate extraction, mycelium is concentrated Jing after Solvent Extract methods,
Crude extract is obtained after gained concentrate is merged；
S3. isolate and purify：The crude extract of step S2 is carried out into silicagel column gradient chromatography, the detection of Jing silica gel thin-layer chromatographies will contain
The elution fraction of target substance proceeds silicagel column, gel filtration chromatography and positive preparative liquid chromatography and isolates and purifies, and obtains mesh
Mark compound；
Wherein, the formula of fungi liquid culture medium described in step S1 is：Contain following component in per liter：Murphy juice 400～
15～25g of 600mL, NaBr, 15～25g of sucrose, 400～600mL of pure water or running water.
3. preparation method according to claim 2, it is characterised in that fermentation described in step S1 is standing for fermentation 2～4 days
Afterwards, the Anuject of final concentration of 0.5～2 mM is added, continues to ferment 18～22 days；Ethyl acetate extraction described in step S2
Number of times is taken for 2～4 times, the organic solvent extraction times are 2～4 times；The organic solvent be chloroform, acetone, ethyl acetate,
In ethanol, methyl alcohol one or more.
4. preparation method according to claim 2, it is characterised in that the concrete grammar isolated and purified described in step S3 is such as
Under：
A. the crude extract Jing silica gel column chromatographies for step S2 being obtained, are carried out pre- with petroleum ether-dichloromethane or petroleum ether-chloroform
Wash-out, carries out gradient elution with chloroform-methanol or methylene chloride-methanol then, and collection compares 100 with effluent volume:0～100:
The component of the wash-out of 1 gradient；Component characteristicses are that solvent is volume ratio 15 on silica gel thin-layer chromatography plate:1 chloroform-methanol
Under expansion, Rf value is 0.29～0.74 component of mixture；
B. the elution fraction that step a is collected is carried out into again silica gel column chromatography, eluent is volume ratio 5:1 petroleum ether-acetone；
C. the elution fraction that step b is collected is carried out into sephadex column chromatography, eluent is methyl alcohol, flow velocity 0.5～0.7
mL/min；
D. the elution fraction that step c is collected is carried out into positive preparative liquid chromatography purifying, eluent is volume ratio 3～5:1 chlorine
Imitative-petroleum ether, in the case of silicagel column fills 20～30g silica gel, the mL/min of flow velocity 2～4, it is 14～28 to collect retention time
The component of min；
E. the elution fraction that step d is collected is proceeded into positive preparative liquid chromatography purifying, eluent is pure chloroform, in silicon
In the case of glue post filling 10～15g silica gel, the mL/min of flow velocity 1～3, the main peak that retention time is 14～35 min is collected, obtained
Gleanings.
5. application of the bromo depsidone class chemicals on antimycotic, antibacterium or desinsection described in claim 1.
6. bromo depsidone class chemicals described in claim 1 are in antifungal agent, antibacterial agent or insecticide is prepared
Using.
7. the application according to claim 5 or 6, it is characterised in that the fungi is Candida albicans.
8. the application according to claim 5 or 6, it is characterised in that the bacterium is Gram-negative bacteria.
9. the application according to claim 5 or 6, it is characterised in that the bacterium is pseudomonas aeruginosa or resistance to methoxyl group
Penicillin staphylococcus aureus.
10. the application according to claim 5 or 6, it is characterised in that the worm is agricultural pests.
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| CN108925565A (en) * | 2018-06-20 | 2018-12-04 | 广东海洋大学深圳研究院 | A kind of application of depsidone class compound |
| CN109593074A (en) * | 2017-09-30 | 2019-04-09 | 华中科技大学 | With the separation preparation and purposes of antibacterial active compounds in secondary fungus metabolite |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109593074A (en) * | 2017-09-30 | 2019-04-09 | 华中科技大学 | With the separation preparation and purposes of antibacterial active compounds in secondary fungus metabolite |
| CN109593074B (en) * | 2017-09-30 | 2020-07-28 | 华中科技大学 | Separation preparation and application of compound with antibacterial activity in fungus secondary metabolite |
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| CN108925565A (en) * | 2018-06-20 | 2018-12-04 | 广东海洋大学深圳研究院 | A kind of application of depsidone class compound |
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