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Marine fungi penicillium crustosum bacterial strain, quinolinone compounds derived from marine fungi penicillium crustosum, and preparation and applications of quinolinone compounds

A technology of quinolinones and compounds, applied in the biological field, can solve problems such as multidrug resistance, loss of efficacy of antibiotics, and inability of antitumor drugs

Inactive Publication Date: 2014-07-23
DALIAN JIAOTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, in clinical treatment, it is found that the drug resistance of tumors is also increasing, and multi-drug resistance appears. The stronger it is, many traditional antibiotics have basically lost their efficacy, so it is necessary to find new anti-tumor drugs and antibiotics to meet the needs of clinical drug development

Method used

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  • Marine fungi penicillium crustosum bacterial strain, quinolinone compounds derived from marine fungi penicillium crustosum, and preparation and applications of quinolinone compounds
  • Marine fungi penicillium crustosum bacterial strain, quinolinone compounds derived from marine fungi penicillium crustosum, and preparation and applications of quinolinone compounds
  • Marine fungi penicillium crustosum bacterial strain, quinolinone compounds derived from marine fungi penicillium crustosum, and preparation and applications of quinolinone compounds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] The preparation method of this quinolinone compound:

[0055] Ⅰ. Fermentation

[0056] Inoculate Penicillium crustosum (Penicillium crustosum) CGMCC No.8516 in the fungal liquid medium for static fermentation, filter, and collect the mycelium and fermentation broth respectively;

[0057] Wherein, the composition of the fungal liquid culture medium contains per liter: 500 mL of potato juice, 500 mL of 40 g / L brine prepared from natural coarse sea salt, and 20 g of sucrose;

[0058] Ⅱ. Rough extraction

[0059] Extracting and concentrating the fermentation broth obtained in step I with ethyl acetate, extracting and concentrating the mycelia obtained in step I with an organic solvent, and combining the obtained concentrates to obtain a crude extract;

[0060] Wherein, the organic solvent is one or more of chloroform, acetone, ethyl acetate, ethanol, methanol;

[0061] Ⅲ. Separation and purification of intermediates

[0062] a. The crude extract obtained in step II was ...

Embodiment 2

[0090] Inhibition of tumor cell growth experiments:

[0091] The human colon cancer cell line HCT116, which is commonly used in anti-tumor drug screening, was used to test the anti-tumor activity in vitro.

[0092] Method: According to the growth rate of the cells, the tumor cells in the logarithmic growth phase were inoculated in a 96-well culture plate, grown adherently for 24 hours, and then the quinolinone compounds to be tested were added. The concentration was set at 500-4 μM, and two replicate wells were set for each concentration. And set the corresponding concentration of normal saline vehicle control and no cell zero well. Tumor cells were stored at 37°C, 5% CO 2 Cultivate under conditions for 24 hours, then discard the culture medium, wash once with phosphate buffer solution and discard, fix the cells with cold methanol, place at 0°C for 10 minutes and then remove the methanol, add 0.25 mg / mL crystal violet staining solution for room temperature staining for 5 min...

Embodiment 3

[0097] Inhibition of Gram-positive bacteria activity test:

[0098] Staphylococcus aureus is a very common germ that sometimes enters the body and causes an infection. The infection can range from mild sores and pimples on the skin to pneumonia or blood infection. Therefore, Staphylococcus aureus is an important target pathogenic microorganism for the development of new antibiotics.

[0099] Method: The international standard filter paper method was used for preliminary screening of antibacterial activity, and the antibacterial activity was measured by the diameter of the inhibition zone (Clinical and Laboratory Standards Institute. 2012. Performance Standards for Antimicrobial Disk Susceptibility Tests; Approved Standard. Eleventh Edition M2–A11 .); rescreening using 96-well plates, using the international standard micro broth dilution method (National Committee for Clinical Laboratory Standards. 2012. Methods for dilution antimicrobial susceptibility tests for bacteria that...

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Abstract

The invention discloses a marine fungi penicillium crustosum bacterial strain, quinolinone compounds derived from marine fungi penicillium crustosum, and a preparation method and applications of the quinolinone compounds. Specific structural formulas of the quinolinone compounds are represented by formula R1-formula R6. According to the preparation method, a culture medium is inoculated with the marine fungi penicillium crustosum bacterial strain AP2T1 CGMCC No.8516 for static fermentation; an obtained fermentation broth is extracted with ethyl acetate, and mycelium is extracted with an organic solvent; obtained crude extracts are mixed, an obtained mixture is subjected to sephadex column chromatography, reverse-phase silica-gel column chromatography, and thin layer chromatography detection using organic solvents; an obtained elution ingredient containing intermediates is subjected to methylation, and thin layer chromatography and reverse-phase silica-gel column chromatography separation and purification so as to obtain target compounds R1-R6. The target compounds R1-R6 possesses wide application prospect in preparation of antitumor drugs, antibacterial agents, and agricultural insecticides.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a strain of marine fungus Penicillium capsid AP2T1, in particular to a quinolinone compound derived from the fungus, a preparation method thereof, and an application thereof in the fields of antitumor drugs, antibacterial antibiotics and insecticides. Background technique [0002] At present, in clinical treatment, it is found that the drug resistance of tumors is also increasing, and multi-drug resistance appears. The stronger it is, many traditional antibiotics have basically lost their efficacy, so it is necessary to find new anti-tumor drugs and antibiotics to meet the needs of clinical drug development. In addition, the excessive use of various chemically synthesized pesticides in agricultural production has caused serious food, soil and water pollution, and endangered human health and ecological balance through the food chain. Pesticide residues have also seriously hinder...

Claims

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Application Information

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IPC IPC(8): C12N1/14C07D215/22C12P17/12A61K31/4704A01N43/42A61P35/00A01P7/00C12R1/80
Inventor 张翼穆军冯妍斯蒂芬妮·格弘德韩金媛温利雪鲍海燕
Owner DALIAN JIAOTONG UNIVERSITY
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