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Method for separating and purifying Pneumocandins B0

A technology of separation and purification, pneumocontin, applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve the problems of complicated purification process and difficult industrialization of extraction and purification, and achieve high purity, mild separation conditions, The effect of improving the quality of the product

Inactive Publication Date: 2014-12-31
BRIGHTGENE BIO MEDICAL TECH (SUZHOU) CO LTD +1
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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 provide a method for separation and purification of pneumocidine B0, which overcomes the defects of difficult industrialization of pneumocidine B0 extraction and purification existing in the prior art, and complicated purification process, and effectively removes the pigment in pneumocidine B0 , improve its purity and content, and make its industrial production easier

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  • Method for separating and purifying Pneumocandins B0

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Take 1000ml of neomocontin B0 fermentation broth, adjust the pH to 4 with oxalic acid, stir for 2 hours, filter, wash the mycelium with 500 ml of water, and collect the mycelium after drying. Take the mycelia, add 1000ml of ethanol solution, soak and stir for 4 hours, filter to obtain about 980ml of extract, concentrate the extract at 50°C to a volume of 100ml; add the mycelium obtained after extraction and filtration to Put into 750ml ethanol solution, soak and stir for 4 hours, filter to obtain about 730ml of extract, concentrate the extract at 50°C to a volume of 75ml, cool to room temperature, combine the two extracts, concentrate at 25°C The ethanol content of the extract to the solution is 25%.

[0031] Take 200ml of AB-8 resin (Cangzhou Baoen Chemical Co., Ltd.) and put it into a resin column (Φ20×500mm), absorb the above-mentioned 25% ethanol content of Neomocontin B0 extract, detect it by HPLC, and then use 400ml Wash the resin column with water, and then wash...

Embodiment 2

[0033] Take 1000ml of neomocontin B0 fermentation broth, adjust the pH to 3 with acetic acid, stir for 1 hour, filter, wash the mycelium with 500 ml of water, and collect the mycelium after drying. Take the mycelium, add 1000ml of methanol solution, soak and stir for 5 hours, filter to obtain about 950ml of extract, concentrate the extract at 50°C to a volume of 100ml; add the mycelium obtained after extraction and filtration to Put into 750ml of methanol solution, soak and stir for 3 hours, filter to obtain about 725ml of extract, concentrate the extract at 50°C to a volume of 72ml, cool to room temperature, combine the two extracts, concentrate at 25°C The methanol content of the leaching liquid to the solution is 20%.

[0034] Take 200ml of HPD-100 resin (Cangzhou Baoen Chemical Co., Ltd.) and put it into a resin column (Φ30×400mm), absorb the above-mentioned Nimocontin B0 extract with a methanol content of 20%, detect it by HPLC, and use it after the adsorption is compl...

Embodiment 3

[0036] Take 1000ml of neomocontin B0 fermentation broth, adjust the pH to 4 with carbonic acid, stir for 2 hours, filter, wash the mycelium with 500 ml of water, and collect the mycelium after drying. Take the mycelium, add 1000ml of acetone solution, soak and stir for 3 hours, filter to obtain about 985ml of extract, concentrate the extract at 50°C to a volume of 100ml; add the mycelium obtained after extraction and filtration to Put into 700ml of acetone solution, stir for 3 hours while soaking, filter to obtain about 735ml of extract solution, concentrate the extract solution at 50°C to a volume of 75ml, cool to room temperature, combine the two extract solutions, and concentrate at 25°C The acetone content of the extract to the solution was 28%.

[0037] Take 200ml of HPD722 resin (Cangzhou Baoen Chemical Co., Ltd.) and put it into a resin column (Φ20×500mm). Adsorb the above-mentioned pneumocidine B0 extract with acetone content of 28%, detect by HPLC, and rinse with 380m...

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Abstract

The invention provides a method for separating and purifying Pneumocandins B0, which comprises the steps of acidifying and extracting a broth, separating a leaching solution by resin adsorption, concentrating, and then drying to obtain the Pneumocandins B0. The obtained product with high purity is a good raw material to prepare antifungal drug Pneumocandins B0. The prepared end product has the advantages of high yield, low production cost, and simple operation, and is especially suitable for industrial production.

Description

technical field [0001] The invention relates to a method for separating and purifying pneumocidine B0. Background technique [0002] Pneumocandin B0 (pneumocandinB0) is a secondary metabolite produced by fungi, and is a precursor compound of the synthetic antifungal drug Caspofungin, with the following structural formula: [0003] [0004] Neomocondine B0 is a precursor compound for the synthesis of caspofungin. If the purity of neomocondine B0 is too low, it will directly affect the yield of each step of the chemical reaction product in the back, and it will bring considerable impact to the refining and purification of the final product caspofungin. Therefore, the preparation of high-purity pneumocidine B0 is the key to the synthesis and purification of caspofungin. [0005] Because pneumocidine B0 is obtained through microbial fermentation, by-products and a large amount of pigments that are similar in structure to pneumocidine B0 will be produced during the fermentati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/56C07K1/36C07K1/22
Inventor 袁建栋
Owner BRIGHTGENE BIO MEDICAL TECH (SUZHOU) CO LTD
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