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Method for preparing astragaloside IV by converting total saponins of astragalus by microorganisms

A technology for the transformation of total astragalosides and microorganisms, which is applied in the direction of drug combinations, pharmaceutical formulas, and medical preparations containing active ingredients, etc., can solve the problems of difficult separation and extraction, low content of astragaloside IV, and overcome saponin damage, regional and Strong stereoselectivity and low cost

Inactive Publication Date: 2012-07-11
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to solve the technical problems of low astragaloside IV content and difficult separation and extraction existing in the prior art, and to provide a method for preparing astragaloside IV through microbial transformation of total astragaloside saponins, especially the use of microorganisms to transform total astragaloside saponins Other saponins in the invention are astragaloside IV, and the method for separating and purifying astragaloside IV from microbial fermentation broth

Method used

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  • Method for preparing astragaloside IV by converting total saponins of astragalus by microorganisms
  • Method for preparing astragaloside IV by converting total saponins of astragalus by microorganisms
  • Method for preparing astragaloside IV by converting total saponins of astragalus by microorganisms

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1: Using Absidia corymbifera Preparation of Astragaloside IV by Conversion of Total Astragalus Saponins

[0028] Two Umbrella Branches Absidia corymbifera ) test tube slant, add 50 mL sterile water to make a spore suspension, inoculate (1 mL per bottle) in a 250 mL shaker flask filled with 50 mL liquid medium, and the liquid medium is potato medium (200 g potato plus water 1 L, boil for half an hour, filter to remove the residue, add 20 g glucose to the filtrate, natural pH). 28°C, 220 rpm, cultured for 24 hours to obtain seed solution. Inoculate 250 mL of seed liquid into a 5 L fermentation bottle containing 1 L of the above liquid medium, culture at 28 °C, 220 rpm for 24 h, add crude total saponins of astragalus (feeding concentration 10 g / L), and transform under the same conditions 72 h. For the results of TLC analysis of the fermentation broth, see figure 1 , Absidia corymbifera It can convert other astragalosides in total astragalosides into astra...

Embodiment 2

[0034] Embodiment 2: Preparation of Astragaloside IV Injection

[0035] Prescription: Astragaloside IV 1.0 g

[0036] Ethanol 200 mL

[0037] Propylene glycol 200 mL

[0038] Glucose 100g

[0039] Make up to 1000 mL with water for injection

[0040] Preparation method: Take the prescribed amount of astragaloside IV, dissolve it in the mixed solution of the prescribed amount of ethanol and propylene glycol at 40°C, stir and add part of the water for injection, then add the prescribed amount of glucose, and make up the water for injection to 1000 mL after completely dissolving , filtered through a 0.22 μm filter, potted, and sterilized to obtain 200 vials of 2 mL specifications of astragaloside IV injection.

[0041]

Embodiment 3

[0042] Example 3 Astragaloside IV Injection Treating Cardiovascular and Cerebrovascular Examples

[0043] 1 Experimental purpose Coxsackie B 3 virus (CVB 3 ) infection-induced viral myocarditis model in mice, evaluate the therapeutic effect of the astragaloside IV injection prepared in the present invention on viral myocarditis, and provide experimental basis for the development and clinical application of new drugs for the treatment of viral myocarditis.

[0044] 2 Experimental method Astragaloside IV was divided into two dose groups, and 50 Balb / c mice were randomly divided into 4 groups. Normal control group (5 rats): no astragaloside IV, no virus inoculation, only the same amount of vehicle; virus control group (15 rats): each rat was intraperitoneally inoculated with 0.1 mL CVB 3 Virus fluid (TCID 50 =10 -7.5 , 10 5 Doubling dilution); administration group I - high dose (15 rats): CVB inoculated with the same method as above 3 Astragaloside IV injection 3 mg / kg by t...

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Abstract

The invention discloses a method for preparing astragaloside IV by converting total saponins of astragalus by microorganisms, which comprises the following steps of: preparing bacteria, mould and yeast into bacterium suspension or spore suspension and inoculating the bacterium suspension or the spore suspension into a seed culture medium to form seed solution; then inoculating the seed solution into a fermentation medium to carry out culturing; after the seed solution grows well, adding the total saponins of astragalus to carry out conversion; carrying out scanning and analysis by a TLC (Thin Layer Chromatography) to convert other astragaloside into the astragaloside IV, so that the amount of the astragaloside IV is improved by over four times; and extracting the conversion solution by an organic solvent and separating and purifying the conversion solution by macroporous resin to obtain a pure product of the astragaloside IV. According to the method disclosed by the invention, the defects of damage to the saponins, serious pollution, poor purposiveness and the like in the process of hydrolyzing the total saponins of astragalus by a chemical method can be overcome and the problem of enzyme instability in the process of hydrolyzing the total saponins of astragalus by an enzyme method can be solved. The method has the advantages of no pollution, strong specificity and high conversion rate. The purity of the prepared astragaloside IV can reach over 99.9 percent.

Description

technical field [0001] The present invention relates to a method for preparing astragaloside IV by microbial transformation, in particular to a method for microbial transformation of astragaloside by using microorganisms containing deacetylase and glycosidase to hydrolyze acetyl groups and sugar groups in total astragaloside saponins to increase the content of astragaloside IV A method for preparing astragaloside IV from total saponins. Background technique [0002] Radix Astragali (Radix Astragali) is one of the commonly used traditional Chinese medicines. Astragalus membranaceus (Fischer) Bunge and Mongolian Astragalus A. membranaceus (Fisch.) Bunge var. mongholicus (Bunge) Dried root of P. K. Hsiao. Astragaloside IV (ASI) is a saponin compound with various pharmacological activities isolated from Astragalus membranaceus, and it is an index component for quality identification of Astragalus membranaceus preparations. Studies have shown that astragaloside IV has a wid...

Claims

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Application Information

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IPC IPC(8): C12P33/20A61K31/7048A61P9/10A61P9/00A61P35/02A61P31/12A61P3/10A61P7/02A61P7/04
Inventor 周珮叶丽史训龙冯美卿李继杨鞠佃文刘晓会
Owner FUDAN UNIV
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