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Preparation method and application of cytochalasin compound Aspochalasin D

A technology of cytochalasin and compounds, applied in the field of medicine, to achieve the effect of low production cost, simple method and easy operation, and inhibition of cell proliferation

Active Publication Date: 2017-09-01
极致生物技术(杭州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there has been no report on the strong inhibitory activity of the compound Aspochalasin D on prostate cancer cells.

Method used

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  • Preparation method and application of cytochalasin compound Aspochalasin D
  • Preparation method and application of cytochalasin compound Aspochalasin D

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Preparation of Compound Aspochalasin D

[0032] (1) Fermentation: Add 40 g of rice and 45 mL of 3% artificial seawater into a 500 mL conical flask, and autoclave at 121° C. for 20 minutes to obtain a rice culture medium.

[0033] The bacterial strain Z-4 was transferred from the slant medium to the rice medium, and cultured at 25° C. for 28 days. After filtering the bacterial solution, extract it with an equal volume of ethyl acetate for 3 times, combine the ethyl acetate layers, and recover the organic solvent under reduced pressure to obtain 40 g of the extract.

[0034] (2) Separation and purification: the above extract was eluted with normal phase silica gel column chromatography (200-300 mesh silica gel, 450 g) with dichloromethane / methanol (volume ratio 100:1-0:100), specifically : Carry out gradient elution with dichloromethane / methanol whose volume ratio is 100:1, 98:2, 95:5, 90:10, 80:20, 50:50, 0:100 in sequence, each concentration volume is 1000mL, thin For...

Embodiment 2

[0038] In Vitro Antitumor Activity Test of Compound Aspochalasin D by MTT Method

[0039] Cell line: prostate cancer PC3 cells;

[0040] Medium: RPMI1640 medium, containing 10% fetal bovine serum;

[0041] Sample preparation: the sample obtained in Example 1 was dissolved in DMSO, and diluted to 5 concentrations according to a certain ratio.

[0042] Activity test experiment: Inoculate 20,000 tumor cells / mL in a 96-well plate with tumor cells in the logarithmic growth phase, add 200 μL of cell suspension to each well, and after culturing for 24 hours, add 1 μL of the above-mentioned prepared drug solution, each concentration set 3 duplicate holes. After the cells were incubated in a 5% carbon dioxide incubator at 37°C for 48 hours, 20 μL of MTT solution with a concentration of 2.5 mg / mL was added, and the culture was continued for 4 hours. The absorbance value (OD value) of each well was measured, and the cell inhibition rate was calculated.

[0043] Table 1 Antitumor acti...

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Abstract

The invention discloses a preparation method and application of a cytochalasin compound Aspochalasin D, and belongs to the technical field of medicines. The preparation method comprises the following steps: inoculating aspergillus flavipes Z-4 of which the preservation number is CCTCC NO: M 2013631 into a rice culture medium, performing fermentation culture, and separating the cytochalasin compound Aspochalasin D from a fermentation liquid. According to the preparation method, the marine fungus, namely the aspergillus flavipes Z-4 is adopted for fermentation culture in the rice culture medium for a first time, the Aspochalasin D is prepared through extraction and separation, and a novel preparation mode is provided for production of the compound; in addition, the preparation method is simple, convenient and easy to operate, low in production cost and applicable to on-scale production; and the Aspochalasin D obtained from a fermentation product of the aspergillus flavipes Z-4 has a remarkable function of inhibiting activity of prostatic cancer cell proliferation, so that the compound can be developed into prostatic cancer prevention medicines or prostatic cancer prevention healthcare food.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a preparation method and application of a cytochalasin compound Aspochalasin D. Background technique [0002] Cancer is a common and frequently-occurring disease that seriously threatens human life and health, and the mortality rate is extremely high. About 4.3 million people die from cancer every year in the world, and about 1 million people die from cancer every year in my country. exacerbated. Therefore, it is of great significance to develop drugs with antitumor activity. [0003] Due to the special living environment such as high pressure, high salt, low temperature, and low nutrition, marine organisms produce and accumulate many secondary metabolites with novel structures and significant biological activities, which have become important resources for the research and discovery of new marine drugs. Compared with other marine biological resources, marine microorganisms wil...

Claims

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Application Information

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IPC IPC(8): C12P17/10C07D209/96A61P35/00C12R1/66
CPCC07D209/96C12P17/10
Inventor 徐金钟王品美丁婉婧李昕阳
Owner 极致生物技术(杭州)有限公司
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