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Purification and preparation method of high-purity Daptomycin

A daptomycin, high-purity technology, applied in the field of purification and preparation of high-purity daptomycin, can solve the problems of complicated operation, large product loss, unreported preparation scale and yield, etc., to improve the purification effect, The effect of improving the purification preparation yield and simplifying the production operation

Active Publication Date: 2010-09-15
CHENGDU YATU BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The patented method repeatedly uses anion exchange chromatography and hydrophobic interaction chromatography alternately, the operation is extremely complicated, the product loss is large, and it is difficult to achieve large-scale preparation
In addition, in all current literature reports on the preparation of high-purity daptomycin, its preparation scale and yield have not been reported.

Method used

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  • Purification and preparation method of high-purity Daptomycin

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Buffer: 5% ammonium acetate solution, pH6.0

[0019] 100 g of crude daptomycin (purity 89.22%) was prepared into a solution with a concentration of 10% by buffer, and filtered to obtain a sample solution.

[0020] Install YT-01 reverse-phase silica gel column 8L (particle size: 150-200 mesh), put the sample solution on the column, first wash it with 3 column volumes of buffer solution, and then add 10% acetonitrile to the same buffer solution for analysis. 2 column volumes, then 2 column volumes with 20% acetonitrile, 2 column volumes with 30% acetonitrile, 1 column volume with 40% acetonitrile, about 40% acetonitrile buffer, Purified daptomycin was eluted from reverse phase silica gel. As determined by HPLC, the purity of daptomycin was 98.63%, and the yield was 54.8%.

Embodiment 2

[0022] Buffer: 8% ammonium acetate solution, adjusted to pH 7.0 with acetic acid

[0023] 100 g of crude daptomycin (purity 89.22%) was prepared into a sample solution with a concentration of 8% using buffer.

[0024] YT-01 reverse phase silica gel (particle size 150-200 mesh): 10L

[0025] Elution condition: 10-40% acetonitrile gradient elution

[0026] The HPLC purity of the purified daptomycin was 98.91%, and the yield was 53.2%.

Embodiment 3

[0028] Buffer: 10% ammonium acetate solution, adjusted to pH 8.0 with acetic acid

[0029] 100 g of crude daptomycin (purity 89.22%) was prepared into a sample solution with a concentration of 5% using a buffer.

[0030] YT-01 reverse phase silica gel (particle size 150-200 mesh): 12L

[0031] Elution condition: 10-40% acetonitrile gradient elution

[0032] The HPLC purity of the purified daptomycin was 99.21%, and the yield was 52.9%.

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Abstract

The invention discloses a purification and preparation method of high-purity Daptomycin, which comprises the following steps of compounding a buffer solution and coarse Daptomycin into a sample solution, absorbing by using composite YT-01reverse phase silica gel columns and carrying out gradient elution or constant elution with the water solution of a strong polar solvent, which is used as a resolution agent. The chromatographic purity is higher than 98%.

Description

technical field [0001] The invention relates to a method for purifying and preparing high-purity daptomycin. Background technique [0002] Daptomycin is a cyclic ester peptide extracted from the fermentation broth of Streptomyces roseosporus. The chemical formula of Daptomycin is C 12 h 10 1N 17 o 25 , the molecular weight is 1620.67, and its structural formula is as follows: [0003] [0004] Compared with the currently clinically used drugs that can treat life-threatening serious infections caused by multidrug-resistant Gram-positive pathogens—vancomycin, linezolid, cobactid, etc., daptomycin has the following advantages: Features: First of all, its mechanism of action is different from all kinds of antibacterial drugs currently on the market. It has the function of destroying bacterial cell membranes and cell walls in many ways, thereby quickly killing pathogenic bacteria and greatly reducing the probability of drug-resistant bacteria. Since its first listing in 2...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61P31/04C07K1/22
Inventor 朱辉韩晓彤邹敬源王元桦张翠英范雪涛黄运昌唐静朱春燕
Owner CHENGDU YATU BIOLOGICAL TECH
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