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Purification method of daptomycin

A technology of daptomycin and a purification method, which is applied in the field of daptomycin purification, can solve the problems of high price, and achieve the effect of easy-to-obtain raw materials and easy industrialization and application

Active Publication Date: 2011-12-14
SHANGHAI LAIYI BIOMEDICAL RES & DEV CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But, used in this method Poros P150 and Poros D50 two kinds of resins, produced by Applied Biosystem Company, its price is very expensive, and although the separation effect is better, because its particle is very fine, it needs a lot of time in the separation and purification process. Large pressure to achieve a certain flow rate

Method used

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  • Purification method of daptomycin
  • Purification method of daptomycin

Examples

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Effect test

Embodiment 1

[0028] The semi-purified daptomycin of embodiment 1

[0029] With reference to the Chinese invention patent application (application number 01805212.6) and its divisional application (application number 200810087401.X) and the method disclosed in U.S. Patent US6696412B1, prepare semi-purified daptomycin, as follows:

[0030] Streptomyces roseospora was fermented under controlled conditions to obtain a daptomycin fermentation liquid. The fermentation broth is centrifuged or filtered to obtain a clear solution. The clarified fermentation broth was applied to anion exchange resin Mitsubishi FP-DA13, then washed with 0.03 mol / L NaCl at pH 6.5, and eluted with 0.3 mol / L NaCl at pH 6.5. Load the eluent onto the reverse phase resin HP-20ss, wash the HP-20ss column with 45% isopropanol, and elute with 55-60% isopropanol, and the purity of the liquid phase is about 90% by HPLC detection semi-purified daptomycin solution.

Embodiment 2

[0031] The daptomycin of embodiment 2 purification

[0032] According to the method of Example 1, a semi-purified daptomycin solution with a liquid phase purity of 90% was obtained, which was loaded onto Sephadex A25, which was pre-balanced with pH 7.0 acetate buffer. After daptomycin is bound to the resin, wash with 3 times column volume of pH 7.0 acetate buffer, and then use 10 times column volume of pH 7.0 acetate buffer containing 0.5mol / L NaCl to elute to obtain purified Daptomycin totomycin. Adopt HPLC to detect the purity of daptomycin, the result is 98.6%, as figure 2 shown.

Embodiment 3

[0033] The daptomycin of embodiment 3 purification

[0034] The semi-purified daptomycin solution with a liquid phase purity of 91% obtained according to the method in Example 1 was loaded onto Sephadex A25, which had previously used a pH 6.0 acetate buffer containing 2 mol / L urea balance. After daptomycin is bound to the resin, wash with 3 times column volume of pH 6.0 acetate buffer containing 2mol / L urea, and then use 10 times column volume of pH 6.0 buffer solution containing 0.2mol / L NaCl and 2mol / L urea Elute with acetic acid buffer, and detect by HPLC to obtain daptomycin with a purity of 98.5%.

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Abstract

The invention discloses a method for purifying daptomuycin. According to the method, half-purified daptomuycin is loaded on a gel type weak anion resin for combination at first and is eluted the next. The purification method provided in the invention enables obtained daptomuycin to have purity higher than 98%; the raw materials used in the method are easily available and cheap; therefore, it is easy to popularize and apply the method.

Description

technical field [0001] The invention relates to the field of medicine purification, in particular to a method for purifying daptomycin. Background technique [0002] Infections caused by gram-positive resistant bacteria are on the rise. Although vancomycin and other glycopeptide antibiotics have a certain therapeutic effect on these drug-resistant pathogens, such as methicillin-resistant Staphylococcus aureus, Staphylococcus epidermidis and Enterococcus resistant strains, The emergence of vancomycin-resistant Gram-resistant bacteria has made this trend more and more important, so there is an urgent need to find a class of antibiotics to combat drug-resistant bacteria. [0003] Daptomycin is an antibiotic produced by Streptomyces roseospora, as figure 1 As shown, it is composed of thirteen amino acids, ten of which form an amino acid peptide ring, and three other amino acids are arranged in a line. Among the three amino acids, the terminal L-tryptophan is followed by a deca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/20C07K1/18
Inventor 夏兴苏永槐罗敏玉孙新强杨志均李秋爽戈梅陈代杰阮林高杨天
Owner SHANGHAI LAIYI BIOMEDICAL RES & DEV CENT
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