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Method for preparing curvularin and indolizidine alkaloid and application

A technology of Curvus sp. and Curvus sp., which is applied in the field of new indolizidine alkaloids and their preparation, can solve the problem that the metabolites of special environment microorganisms do not attract enough attention, and achieves short cycle, simple process, Effects of strong acetylcholinesterase inhibitor activity

Inactive Publication Date: 2012-01-25
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Although the secondary metabolites of microorganisms have been widely studied, the metabolites of some special environment microorganisms have not attracted enough attention

Method used

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  • Method for preparing curvularin and indolizidine alkaloid and application
  • Method for preparing curvularin and indolizidine alkaloid and application
  • Method for preparing curvularin and indolizidine alkaloid and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Curvularia IFB-Z10 of marine origin ( Curvularia sp . ) separation and purification

[0029] Dissect the white meager under aseptic conditions, disinfect the surface of the taken out meager intestines in 75% alcohol for 2 minutes, rinse with sterile water for 3 times, add a small amount of sterile water and grind them in a sterile mortar. Gradiently dilute the polishing solution with sterile water to 10 -1 , 10 -2 , 10 -3 , 10 -4 , respectively take 0.2 ml of each gradient dilution and spread it on Chase medium (sucrose 30 g, NaNO 3 3.0 g, NaCl 0.5 g, K 2 HPO 4 1.0 g, MgSO 4 ·7H 2 O 0.5 g, FeSO 4 ·7H 2 O 0.01 g, 1 L of distilled water), cultured in a 28°C incubator, and after the colonies grew out, the mycelium was picked and purified to obtain the intestinal fungus IFB-Z10 of white meager fish, which was tested by molecular biology and morphology identified as Curvularia IFB-Z10 ( Curvularia sp . ), is now deposited in the China Center for...

Embodiment 2

[0030] Embodiment 2: Curvularia IFB-Z10 of marine origin ( Curvularia sp.) liquid fermentation

[0031] Activation of marine-derived Curvularia IFB-Z10 ( Curvularia sp.), inoculate fresh bacterium blocks into 1000 ml Erlenmeyer flasks, each bottle contains 400 ml of modified Chapeauer medium, inoculate 5-6 bottles on a shaker, at 150 rpm, 28-30 ℃ Cultivate for 3 days as the seed solution, then inoculate the seed solution with 20 ml inoculum in a 1000 ml Erlenmeyer flask containing 400 ml of modified Chapeauer's medium, and ferment for 12 days at 150 rpm and 28-30 °C.

Embodiment 3

[0032] Embodiment 3: Extraction and separation of indolizidine alkaloids

[0033] The fermented liquid obtained in Example 2 was filtered through gauze, the filtrate was extracted with ethyl acetate, concentrated and dried in vacuo to obtain black crude extract F1. Carry out silica gel column chromatography on extract F1, use 4 times the volume of chloroform successively, chloroform:methanol=100:1, chloroform:methanol=100:2, chloroform:methanol=100:4, carry out elution, concentrate 100:1 Segment obtains black extract F2; 100:4 segment obtains black extract F3; Then Sephadex LH-20 separates extract F2 to obtain curvulamine A, and merges other fractions to obtain brown extract F2-1; extract F2- 1 with high pressure liquid chromatography (column: Allsphere ODS 5 mu m (250×4.6 mm) column; mobile phase: methanol / water=80 / 20, flow rate: 2.0 mL / min) to separate curvulamine B (retention time t R =26.1 min); curvulamine C (retention time t R =22.3 min); for the extract F3, use hi...

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Abstract

The invention belongs to the technical field of biological pharmacy and particularly relates to indolizidine alkaloid extracted from marine fungi liquid fermentations, and a preparation method and an application thereof. The indolizidine alkaloid is obtained through separating and purifying marine curvularin IFB-Z10 (curvularia sp.) fermentation liquid. The alkaloid has higher inhibiting effect on acetylcholine esterase and does not have toxicity on cells, so the indolizidine alkaloid can be used as a lead compound of acetylcholine esterase inhibitors, is applied to the acetylcholine esterase inhibitor preparation and can be prepared into medicine for treating Alzheimer's diseases.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, in particular to the marine fungus IFB-Z10 ( Curvularia sp . A class of new indolizidine alkaloids extracted from the liquid fermented product of ) and its preparation method and application. Background technique [0002] Although the secondary metabolites of microorganisms have been widely studied, the metabolites of some special environment microorganisms have not attracted enough attention. The particularity of the marine environment has created the unique metabolic mode and metabolites of marine fungi. In recent years, people have been looking for antibiotics with new mechanisms of action from fungi in various marine environments. In addition to antibiotics, marine fungi can also produce inhibitors of various enzymes, and the diversity of marine fungi provides a rich source of secondary metabolites. [0003] Alzheimer's disease (AD) is a neurodegenerative disease that seriously...

Claims

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Application Information

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IPC IPC(8): C12N1/14C07D471/22C07D471/04A61K31/55A61K31/437A61P25/28C12R1/645
Inventor 谭仁祥张高飞宋勇春崔江涛汪伟
Owner NANJING UNIV
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