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Echinocandin compound purifying method

A technique for echinocandin and purification method, which is applied in the fields of peptide preparation, chemical instruments and methods, organic chemistry, etc., to achieve the effect of simple method and high sample purity

Inactive Publication Date: 2014-01-01
ZHEJIANG HISUN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No reports have disclosed the isolation of WF11899A using this method

Method used

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  • Echinocandin compound purifying method
  • Echinocandin compound purifying method
  • Echinocandin compound purifying method

Examples

Experimental program
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Effect test

Embodiment 1

[0038] 30 liters of WF11899A fermented broth was added to filter with diatomaceous earth to obtain mycelia. The mycelium is soaked with 40 liters of ethanol, filtered to obtain 35 liters of soaking liquid, concentrated under reduced pressure to obtain 10 liters of concentrated liquid, which contains about 30 grams of WF11899A. The concentrated solution was filtered through a 0.45um microporous membrane to obtain a clear prepurified solution. The prepurified solution was adsorbed with 2.5 liters of HP20SS resin. After the adsorption is completed, rinse the resin with 5 liters of water, and then wash it with 8 liters of 30-40% (V / V) ethanol. Finally, it was eluted with 9 liters of 55% (V / V) ethanol, and the qualified eluate was collected. The obtained eluate contained about 23.3 grams of WF11899A, and the chromatographic purity was 75%.

Embodiment 2

[0040] 11 liters of clear liquid containing 27.3 grams of WF11899A was processed according to the method of Example 1, and the pH was adjusted to 7.5 with 0.5 mol / l sodium hydroxide solution to form a prepurified solution of WF11899A sodium salt. The pre-purified solution was adsorbed with 2 liters of HP20 resin, and after the adsorption was completed, the resin was washed with 4 liters of water, and then washed with 6 liters of 30% (V / V) ethanol. Finally, it was eluted with 6.5 liters of 60% (V / V) ethanol, and 2.4 liters of the eluate was collected. The resulting eluate contained about 19.3 grams of WF11899A. The resulting eluate had a chromatographic purity of 70%.

Embodiment 3

[0042] The obtained 9 liters of clarified liquid containing 28 grams of WF11899A were processed according to the method of Example 1, and the pH was adjusted to 7.5 with 5% (w / w) ammonia solution to form a pre-purified solution of WF11899A ammonium salt. The pre-purified solution was adsorbed with 2.5 liters of SP825 resin, and after the adsorption was completed, the resin was washed with 5 liters of water, and then washed with 7 liters of 30% (V / V) methanol. Finally, it was eluted with 10 liters of 50% (V / V) methanol, and 3.6 liters of the eluate was collected, which contained about 22.7 grams of WF11899A in the obtained eluate. The resulting eluate had a chromatographic purity of 72%.

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Abstract

The invention relates to an echinocandin compound purifying method, and especially relates to a WF11899A purifying method. The method comprises the processes of pre-purifying solution preparation, resin chromatography separation, crystallization and the like. The method has the advantages of simplicity, no use of a mixed solvent, high sample purity and the like.

Description

technical field [0001] The invention relates to a purification method of echinocandin compounds, in particular to a purification method of WF11899A. Background technique [0002] Echinocandin compounds have antifungal effects, including echinocandin B, WF11899A, echinocandin C, pulmonary colistin A0, pulmonary colistin B0, etc. Such compounds can be further prepared to obtain semi-synthetic compounds, such as caspofungin, micafungin, anidifungin and the like. Echinocandin compounds are obtained by microbial fermentation and contain a large number of by-products. These by-products are often difficult to remove in subsequent separation and purification. [0003] US5021403 discloses a chromatographic purification method for echinocandin compounds, which includes repeated resin adsorption and silica gel chromatography, and the process is complicated. WO2008048627 discloses a method for purifying echinocandins, including extraction, washing, precipitation and the like. During...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/56C07K1/16
Inventor 杨华剑梁陈宏周永健陈峰王玲萍郑玲辉白骅
Owner ZHEJIANG HISUN PHARMA CO LTD
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