CN107954860B - Succinic acid derivative and preparation method and application thereof - Google Patents
Succinic acid derivative and preparation method and application thereof Download PDFInfo
- Publication number
- CN107954860B CN107954860B CN201711007916.XA CN201711007916A CN107954860B CN 107954860 B CN107954860 B CN 107954860B CN 201711007916 A CN201711007916 A CN 201711007916A CN 107954860 B CN107954860 B CN 107954860B
- Authority
- CN
- China
- Prior art keywords
- succinic acid
- acid derivative
- preparation
- extract
- methanol
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C59/00—Compounds having carboxyl groups bound to acyclic carbon atoms and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups
- C07C59/40—Unsaturated compounds
- C07C59/76—Unsaturated compounds containing keto groups
-
- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/44—Polycarboxylic acids
- C12P7/50—Polycarboxylic acids having keto groups, e.g. 2-ketoglutaric acid
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention discloses a succinic acid derivative and its preparation and application, characterized in that the succinic acid derivative has a structural formula shown in I, the preparation method comprises the steps of obtaining a fermentation product of a novel succinic acid derivative by fermenting and culturing marine trichoderma with the preservation number of CGMCC No:12969, extracting the fermentation liquor with ethyl acetate to obtain a crude extract, and separating and purifying the crude extract by reduced pressure silica gel column chromatography and reversed phase semi-preparative high performance liquid chromatography.
Description
Technical Field
The invention relates to a succinic acid derivative, in particular to a method for preparing the succinic acid derivative by using marine trichoderma and application of the compound in preparing candida albicans inhibitors.
Background
Candida albicans (C., (C.))Candida albicans) Is a normal flora colonizing the normal human digestive tract, respiratory tract and mucous membrane of female genital tract, and is also one of the most common opportunistic pathogens in candida. In recent years, due to the large application of immunosuppressive agents, tumor radiotherapy and chemotherapyThe development of therapies, invasive treatments, organ transplants, and the widespread use of large amounts of broad-spectrum antibiotics has led to a dysbacteriosis of the normal flora, which has led to a significant increase in the infection and incidence of candida albicans compared to previous treatments. Because the existing antifungal drugs are few and have strong toxic and side effects, drug-resistant strains continuously appear, and the occurrence of biomembrane-related infection makes the search of new antifungal drugs a research hotspot in the field of microbiology. The marine microorganism is a potential antifungal compound source, is a research hotspot in recent years, and has the advantages of easy culture, short fermentation period and the like. The inventor researches to obtain the marine trichodermaTrichodermasp. (preserved in China general microbiological culture Collection center with the preservation number of CGMCC No: 12969), the liquid fermented ethyl acetate extract has better anti-Candida albicans activity, so that the active ingredients of the extract are researched. At present, the chemical structure and the anti-candida albicans activity of the compound are not reported, so that the related medicines are not seen in the market.
Disclosure of Invention
The invention aims to solve the technical problem of providing a succinic acid derivative with anti-candida albicans activity, and a preparation method and application thereof.
The technical scheme adopted by the invention for solving the technical problems is as follows: a succinic acid derivative having the formula I:
the preparation method of the succinic acid derivative specifically comprises the following steps:
(1) fermentation production
Trichoderma with preservation number of CGMCC No. 12969Trichodermasp. streak revival, transferring to PDB culture medium, culturing at 25 deg.C and 180r/min for 4 days to obtain seed culture solution; then inoculating the seed solution into a PDB culture medium according to the inoculation amount of 10% of the volume percentage, and fermenting for 14 days at 25 ℃ and 150rpm in a shaking table to obtain fermentation liquor;
(2) obtaining extract
Extracting the fermentation liquor with ethyl acetate of the same volume for 3 times, and concentrating the extract under reduced pressure to remove ethyl acetate to obtain crude extract;
(3) separation and purification of Compound
Dissolving the crude extract by using a mixed solvent of dichloromethane and methanol, adding 200-mesh silica gel of 300 meshes for mixing, eluting by using petroleum ether/ethyl acetate with a gradient of 10:1 in volume ratio as an eluent, performing reduced pressure silica gel column chromatography on the crude extract, collecting an elution component, and performing separation and purification by using semi-preparative reverse phase high performance liquid chromatography to obtain the succinic acid derivative, wherein the structure of the succinic acid derivative is shown as a formula I:
the preparation method of the PDB culture medium comprises the following steps: dissolving potato extract 2g and glucose 8g in 400mL seawater.
The volume ratio of the dichloromethane to the methanol in the dichloromethane and methanol mixed solvent is 1: 1.
the eluent of the semi-preparative reverse phase high performance liquid chromatography is formed by mixing acetonitrile and water according to the volume ratio of 25: 75.
The application of the succinic acid derivative and the application of the succinic acid derivative in the aspect of preparing the candida albicans inhibitor.
Compared with the prior art, the invention has the advantages that: the invention relates to a succinic acid derivative and a preparation method and application thereof, wherein a fermentation product containing a novel succinic acid derivative is obtained through microbial fermentation culture, then the fermentation product is soaked in methanol and extracted by ethyl acetate to obtain a crude extract, and the crude extract is separated and purified by reduced pressure silica gel column chromatography and reversed-phase semi-preparative high performance liquid chromatography.
The Trichoderma (Trichoderma sp.) is HPQJ34 strain with the preservation number of CGMCC number 12969, is preserved in China general microbiological culture Collection center at 10 and 13 days in 2016, and has the preservation address of No. 3 Hokkaido 1, China academy of sciences, North Cheng Yang district, Beijing.
Detailed Description
The present invention will be described in further detail with reference to examples.
Example 1
A succinic acid derivative has the structural formula shown in the specification:
example 2
The preparation method of the succinic acid derivative shown as the formula I specifically comprises the following steps:
(1) fermentation production
Trichoderma with preservation number of CGMCC No. 12969Trichodermasp. streak revival, transferring the obtained product into a 250mL conical flask filled with 100 mL of PDB culture medium, and culturing the product at 25 ℃ and 180r/min for 4 days to obtain a seed culture solution; then inoculating the seed solution into a 1000mL conical flask filled with a PDB culture medium (2 g of potato extract and 8g of glucose dissolved in 400mL of seawater) according to the inoculation amount of 10 percent by volume, and fermenting for 14 days at 25 ℃ and 150rpm in a shaking table to obtain a fermentation liquid;
(2) obtaining extract
Extracting the fermentation liquor for 3 times by using ethyl acetate with the same volume, and concentrating the extract liquor under reduced pressure to remove the ethyl acetate to obtain 17 g of crude extract;
(3) separation and purification of Compound
Dissolving the crude extract by using a mixed solvent of dichloromethane and methanol, adding 200-mesh silica gel of 300 meshes for mixing, eluting by using petroleum ether/ethyl acetate with a gradient of 10:1 in volume ratio as an eluent, performing reduced pressure silica gel column chromatography on the crude extract, collecting an elution component, and performing separation and purification by using semi-preparative reverse phase high performance liquid chromatography to obtain the succinic acid derivative, wherein the structure of the succinic acid derivative is shown as a formula I:
the compound I, white powder, formula C11H16O5Cationic HRESIMSm/z：246.1336 [M + H]+，1H and13the C-NMR data are shown in Table 1.
TABLE 1 preparation of compound I1H and13c NMR data (600 and 150MHz, in CDCl)3）
Example 3
In vitro testing of Candida albicans resistance (96-well plate bacteriostasis method)
(1) Experimental sample
Preparing a solution of a sample to be detected: the test sample is the purified compound I isolated and purified in the above example 1, and an appropriate amount of the sample is precisely weighed and prepared into a solution with a required concentration by DMSO for activity measurement.
(2) Experimental methods
The 96-well plate anti-candida albicans test method comprises the following steps: the test compound was diluted step by step in 20% DMSO/saline and transferred 10 μ L to a 96-well flat-bottom microplate. Candida albicans strain was cultured in Sabouraud agar medium (SDA) at 30 ℃ for 16-20 hours under aerobic conditions. A series of compounds at different concentrations were added to RPMI1640 medium and inoculated with Candida albicans and incubated at 35 ℃ for 46 hours. Fluconazole at 8. mu.g/ml was used as a positive control, and DMSO solution was used as a negative control. The concentrations of the test compounds were 1. mu.g/ml, 2. mu.g/ml, 4. mu.g/ml and 8. mu.g/ml, respectively. After completion of the culture, the absorbance at 600nm was measured, and the inhibition ratio was calculated according to the following formula. Inhibition (%) = (OD)R-OD)/(ODR-ODB)
ODR: the light absorption value of the bacteria liquid control hole; ODB: blank light absorption values; OD: the samples were assayed for absorbance in the wells.
(3) Results of the experiment
The results of the inhibition of Candida albicans by different concentrations of Compound I in the 96-well plate antifungal assay are shown in Table 2.
TABLE 2 inhibition of Candida albicans by Compound I at various concentrations (%)
The above table shows that the compound I has significant anti-Candida albicans effect, and can be used as a Candida albicans inhibitor or research of other antifungal drug lead compounds.
The above description is not intended to limit the present invention, and the present invention is not limited to the above examples. Those skilled in the art should also realize that such changes, modifications, additions and substitutions are within the true spirit of the invention.
Claims (3)
1. A succinic acid derivative characterized by: the succinic acid derivative has a structural formula shown as I:
2. a process for the preparation of derivatives of succinic acid according to claim 1, characterized in that it comprises the following steps:
(1) fermentation production
Carrying out streak rejuvenation on Trichoderma sp with the preservation number of CGMCC No. 12969, transferring into a PDB culture medium, and culturing at 25 ℃ and 180r/min for 4 days to obtain a seed culture solution; then inoculating the seed solution into a PDB culture medium according to the inoculation amount of 10 percent of volume percentage, and fermenting for 14 days in a shaking table at 25 ℃ and 150rpm to obtain a fermentation liquid, wherein the preparation method of the PDB culture medium comprises the following steps: dissolving 2g of potato extract and 8g of glucose in 400mL of seawater to prepare the potato extract;
(2) obtaining extract
Extracting the fermentation liquor with ethyl acetate of the same volume for 3 times, and concentrating the extract under reduced pressure to remove ethyl acetate to obtain crude extract;
(3) separation and purification of Compound
Dissolving the crude extract by using a mixed solvent of dichloromethane and methanol, adding 200-mesh silica gel of 300 meshes for mixing, eluting by using petroleum ether/ethyl acetate with a gradient of 10:1 in volume ratio as an eluent, performing reduced pressure silica gel column chromatography on the crude extract, collecting an elution component, and performing separation and purification by using semi-preparative reverse phase high performance liquid chromatography to obtain the succinic acid derivative, wherein the structure of the succinic acid derivative is shown as a formula I:
3. Use of a succinic acid derivative according to any one of claims 1-2, wherein: the succinic acid derivative is applied to the preparation of candida albicans inhibitors.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711007916.XA CN107954860B (en) | 2017-10-25 | 2017-10-25 | Succinic acid derivative and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711007916.XA CN107954860B (en) | 2017-10-25 | 2017-10-25 | Succinic acid derivative and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107954860A CN107954860A (en) | 2018-04-24 |
| CN107954860B true CN107954860B (en) | 2020-08-11 |
Family
ID=61964088
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711007916.XA Active CN107954860B (en) | 2017-10-25 | 2017-10-25 | Succinic acid derivative and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107954860B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106518818A (en) * | 2016-10-14 | 2017-03-22 | 宁波大学 | Furanone compound and preparation method and application thereof |
| CN106966886A (en) * | 2017-01-24 | 2017-07-21 | 宁波大学 | A kind of cyclopentene ketone compounds and its preparation method and application |
| CN107973803A (en) * | 2017-11-02 | 2018-05-01 | 宁波大学 | A kind of seven yuan of lactone benzofuran derivs and its preparation method and application |
-
2017
- 2017-10-25 CN CN201711007916.XA patent/CN107954860B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106518818A (en) * | 2016-10-14 | 2017-03-22 | 宁波大学 | Furanone compound and preparation method and application thereof |
| CN106966886A (en) * | 2017-01-24 | 2017-07-21 | 宁波大学 | A kind of cyclopentene ketone compounds and its preparation method and application |
| CN107973803A (en) * | 2017-11-02 | 2018-05-01 | 宁波大学 | A kind of seven yuan of lactone benzofuran derivs and its preparation method and application |
Non-Patent Citations (4)
| Title |
|---|
| Antibacterial Secondary Metabolites from an Endophytic Fungus, Eupenicillium sp. LG41;Gang Li et al.;《J. Nat. Prod. 》;20141030;第77卷(第11期);第2335-2341页 * |
| Asperitaconic acids A–C, antibacterial itaconic acid derivatives produced by a marine-derived fungus of the genus Aspergillus;Lijian Ding et al.;《The Journal of Antibiotics》;20180704;第71卷(第10期);第902-904页 * |
| Enzymatic Resolution of α-Methyleneparaconic Acids and Evaluation of their Biological Activity;Kuheli Chakrabarty et al.;《Chirality》;20150109;第27卷(第3期);第239-246页 * |
| Tensyuic Acids, New Antibiotics Produced by Aspergillus niger FKI-2342;Yoko HASEGAWA et al.;《Chem. Pharm. Bull.》;20070703;第55卷(第9期);第1338-1341页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107954860A (en) | 2018-04-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110527629B (en) | Marine fungus-derived brefeldin A, preparation method and application thereof in resisting agricultural pathogenic bacteria | |
| CN110863021B (en) | Preparation method and application of cytochalasin compound | |
| CN108315264A (en) | A kind of polyketide in sea paint endogenetic fungus source and its application in preparing anti-inflammatory drug | |
| CN111334438A (en) | Preparation method of aromatic butenolide dimer and application of aromatic butenolide dimer in preparation of antibacterial drugs | |
| CN106967024A (en) | A kind of α pyrone derivatives and its preparation method and application | |
| CN110330544B (en) | 4,4, 1-bicyclic steroid compound and preparation method and application thereof | |
| CN111072670B (en) | Diketopiperazine compound and preparation method and application thereof | |
| CN107973803B (en) | Seven-membered lactonofuran derivative and preparation method and application thereof | |
| CN110003153B (en) | Benzofuran compound and preparation method and application thereof | |
| CN108707090B (en) | Chlorine-containing aromatic compound and preparation method and application thereof | |
| CN110357788B (en) | Polyketone compound and preparation method and application thereof | |
| CN107805188B (en) | Biphenyl compound and preparation method and application thereof | |
| CN107954860B (en) | Succinic acid derivative and preparation method and application thereof | |
| CN110002996B (en) | Diphenyl ether compound and preparation method and application thereof | |
| CN114380764B (en) | Thiazoline siderophore compound and preparation method and application thereof | |
| CN113444131B (en) | N-acetylglucosamine compounds, and preparation method and application thereof | |
| CN108794502B (en) | Trichothecene compound and preparation method and application thereof | |
| CN108727169A (en) | A kind of preparation method of the biphenyl ether compound in marine fungi source and the application as antiseptic | |
| CN115109023A (en) | Macrolide compound FWYZ52-A, and fermentation strain, fermentation method and application thereof | |
| CN114621092A (en) | Phenolic compound in mangrove plant-derived fungi and preparation method thereof | |
| CN108441427B (en) | Arthriospora fungi and pyridone alkaloid compound produced by same | |
| CN114469908B (en) | Preparation method and application of acinetobacter baumanii-resistant compound stephol | |
| CN112107602B (en) | A pair of twin-nitrogen containing alkaloid enantiomers, preparation and application thereof | |
| CN111285828B (en) | Compound proximicin and preparation method and application thereof | |
| CN116836140B (en) | Compound with anti-inflammatory activity and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |