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Preparation method of high-purity pneumocandin B0

A lung capsule Kangding, high-purity technology, applied in the preparation methods of peptides, chemical instruments and methods, organic chemistry and other directions, can solve the problems of unsuitability for industrial production, cumbersome processes, and high costs, and achieve shortened process cycles. The effect of improving quality

Active Publication Date: 2013-05-01
NCPC NEW DRUG RES & DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chinese patent CN101659693A discloses a kind of preparation lung capsule Kangding B 0 The method, this method repeatedly extracts Lung Sac Kangding B with two kinds of solvents 0 , the process is cumbersome, the yield is low, and the total extraction yield is less than 50%; and this method uses acidic alumina to decolorize the extract, which has high cost and is not suitable for industrial production

Method used

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  • Preparation method of high-purity pneumocandin B0
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  • Preparation method of high-purity pneumocandin B0

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Take Lung Sac Kangding B 0 Fermentation broth 10.0L, fermentation broth unit 832μg / mL. 100 g of perlite was added to the fermentation broth, stirred for 30 minutes and then vacuum filtered to obtain 2.0 kg of fermentation culture. Add 8L of industrial ethanol to the fermentation culture, stir mechanically for 1 hour, and then vacuum filter to collect the extract. The extract was decolorized through a D303 resin column with a diameter-to-height ratio of 1:6, the resin loading capacity was 500 mL, and the flow rate was 1000 mL / h. Collect the decolorized solution, adjust the concentration of the solvent to 30%, and introduce the HZ830 resin column with a diameter-to-height ratio of 1:6 for adsorption. The resin loading capacity is 400 mL, and the flow rate is 800 mL / h. The saturated resin was purified and washed with 40% ethanol, and then desorbed with 80% ethanol at a flow rate of 200mL / h. The desorption solution was concentrated, extracted with ethyl acetate, and acet...

Embodiment 2

[0041] Take Lung Sac Kangding B 0 Fermentation broth 10.0L, fermentation broth unit 762μg / mL. Add 300g of perlite to the fermentation broth, stir for 1 hour and then vacuum filter to obtain 2.2kg of fermentation culture. Add 13.2L of industrial ethanol to the fermentation culture, stir mechanically for 2 hours, and then vacuum filter to collect the extract. The extract was decolorized through a D303 resin column with a diameter-to-height ratio of 1:6, the resin loading capacity was 500 mL, and the flow rate was 1000 mL / h. Collect the decolorized solution, adjust the concentration of the solvent to 40%, and introduce the HZ830 resin column with a diameter-to-height ratio of 1:6 for adsorption. The resin loading capacity is 400 mL, and the flow rate is 800 mL / h. The saturated resin was purified and washed with 50% ethanol, and then desorbed with 90% ethanol at a flow rate of 200 mL / h. The desorption solution was concentrated, extracted with ethyl acetate, and acetone was crys...

Embodiment 3

[0043] Take Lung Sac Kangding B 0 Fermentation broth 20.0L, fermentation broth unit 973μg / mL. Add 500 g of perlite to the fermentation broth, stir for 1 hour and then vacuum filter to obtain 4.5 kg of fermentation culture. Add 19L of industrial ethanol to the fermentation culture, mechanically stir for 1 hour, and then vacuum filter to collect the extract. The extract was decolorized through a D303 resin column with a diameter-to-height ratio of 1:6, the resin loading capacity was 1000 mL, and the flow rate was 2000 mL / h. Collect the decolorized solution, adjust the concentration of the solvent to 30%, and introduce the HZ830 resin column with a diameter-to-height ratio of 1:6 for adsorption. The resin loading is 1000 mL, and the flow rate is 2000 mL / h. The saturated resin was purified and washed with 40% ethanol, and then desorbed with 90% ethanol, and the desorption flow rate was 500mL / h. The desorption solution was concentrated, extracted with ethyl acetate, and crystall...

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Abstract

The invention discloses a method for preparing pneumocandin B0 by using a fermentation culture product of filamentous fungi glarea lozoyensis. According to the method, a filter aid is adopted; with steps such as leaching, discoloring, adsorption, crystallization, and the like, a pneumocandin B0 crude extract is obtained; and the crude extract is subjected to chromatographic separation by using polymer microspheres, such that the high-purity pneumocandin B0 product is obtained. The method provided by the invention has the advantages that: pneumocandin B0 is extracted and separated by using macroporous resin; the process is simple and feasible, and is suitable for industrialized productions; the polymer microspheres are used in chromatographic separation of the product for a first time, and high-purity pneumocandin B0 product can be prepared.

Description

technical field [0001] The invention belongs to the technical field of industrial microorganisms, and in particular relates to a high-purity Lung Sac Kangding B 0 method of preparation. Background technique [0002] Echinocandin is a new type of antifungal drug, which is effective against Candida and Aspergillus, and has high safety. Currently there are 3 varieties on the market, namely caspofungin from Merck, micafungin from Fujisawa Pharmaceutical Co., Ltd., and anifungin from Pfizer. Among them, caspofungin is an echinocandin approved by the U.S. FDA for the first time. It was developed by Merck and was first launched in the U.S. in February 2001 for the infection of Candida albicans and Aspergillus fumigatus. [0003] Caspofungin is the first echinocandin antifungal drug approved for the treatment of invasive aspergillosis due to intolerance or poor response to other antifungal drugs (Groll AH, Piscitelli SC, Walsh TJ. Adv Pharmacol, 1998, 44:343-500). Caspofungin ha...

Claims

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Application Information

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IPC IPC(8): C07K7/56C07K1/16
Inventor 李晓露张雪霞任风芝林毅林旸王海燕李宁王健陈书红李丽红成晓迅高月麒张金娟张丽蒋沁段宝玲
Owner NCPC NEW DRUG RES & DEV
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