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Daptomycin extraction method

A technology of daptomycin and its extraction method, which is applied to the preparation method of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problems of difficult removal of pigments and impurities in fermentation liquid, complicated operation steps, and deep color of products, and achieve The effect of reduced use of acid-base adjustment reagents, high purity, and excellent raw materials

Active Publication Date: 2016-04-13
LIVZON GROUP FUZHOU FUXING PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extraction and purification method of daptomycin disclosed in this document has complex operation steps, difficult removal of pigments and impurities in the fermentation liquid, dark color and low purity of the final product, and the different fermentation liquids produced by fermentation of different strains may not be able to with the same extraction method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Step (1): 30L of daptomycin fermentation broth, the content of daptomycin is 2000mg / L, filter with ceramic membrane, concentrate to half the volume, add drinking water to top wash while filtering, keep the amount of water added and the flow rate of filtration Consistent, until the total filtrate volume is 5 times the fermentation broth volume, stop filtering. 160 L of filtrate was obtained, the content of daptomycin was 340 mg / L, and the yield was 95.5%.

[0039] Step (2): The filtrate obtained in step (1) is adsorbed through a macroporous adsorption resin HZ801 column (macroporous adsorption resin HZ801 column was purchased from Shanghai Huazhen Technology Co., Ltd.) at a flow rate of 0.8BV / h. After adsorption, wash with water until the leaked liquid Nearly colorless, pre-washed with 20% ethanol, eluted with 60% ethanol resin to collect the eluate with daptomycin units greater than 100 mg / L. 9 L of the eluent was obtained, the content of daptomycin was 5700 mg / L, and ...

Embodiment 2

[0045] The fermented liquid is 30L, the content of daptomycin is 2000mg / L, filter with ceramic membrane, concentrate to half of the volume, add drinking water to top wash while filtering, keep the amount of water added and the flow rate of filtration consistent, and continue until the total volume of the filtrate is the fermentation When the amount is 2 times of the liquid volume, stop filtering.

[0046] The filtrate is adsorbed through a macroporous adsorption resin HZ801 column, with a flow rate of 0.8BV / h. After the adsorption, it is washed with water until the leakage liquid is nearly colorless, pre-washed with 10% ethanol, and eluted with 80% ethanol to collect the daptomycin unit greater than 100mg / L. eluent. Get the eluate.

[0047] After the eluate is ultrafiltered with a 5,000 Dalton ultrafiltration membrane, it is concentrated by nanofiltration, and the temperature during ultrafiltration and filtration is controlled below 20 degrees. Concentrate to about 5,000 uni...

Embodiment 3

[0052] The fermented liquid is 30L, the content of daptomycin is 2000mg / L, filter with ceramic membrane, concentrate to half of the volume, add drinking water to top wash while filtering, keep the amount of water added and the flow rate of filtration consistent, and continue until the total volume of the filtrate is the fermentation When the amount is 2 times of the liquid volume, stop filtering.

[0053] The filtrate is adsorbed through the macroporous adsorption resin HZ801 column, the flow rate is 0.8BV / h, after the adsorption, it is washed with water until the leakage liquid is almost colorless, pre-washed with 30% ethanol, and eluted with 50% ethanol to collect the daptomycin unit greater than 100mg / L eluent. Get the eluate.

[0054] After the eluate is ultrafiltered with a 50,000 Dalton ultrafiltration membrane, it is concentrated by nanofiltration, and the temperature during ultrafiltration and filtration is controlled below 20 degrees. Concentrate to about 20,000 uni...

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Abstract

The invention relates to the technical field of antibiotic production, in particular to a daptomycin extraction method. The method includes the following steps of firstly, filtering daptomycin fermentation liquor through a ceramic film; secondly, making filtered liquor pass through large-pore adsorption resin so that eluant can be collected; thirdly, mixing NM-Q anion exchange resin with the obtained eluant, and collecting eluant; fourthly, making the eluant obtained in the third step pass through a reverse phase silica gel MNsil-C18 column, conducting gradient elution on the column through an ammonium acetate ethanol solution, and collecting eluant; fifthly, concentrating the daptomycin eluant obtained in the fourth step, adding calcium acetate and isopropyl alcohol, and conducting standing, crystallization, suction filtration and drying to obtain daptomycin crystallization powder. The method has the advantages that the virulent acetonitrile solution is not used as the eluant, pressure on the environment and health of production personnel is reduced, the use of acid-base regulator is reduced, and the production cost is reduced.

Description

technical field [0001] The invention relates to the technical field of antibiotic production, in particular to a method for extracting daptomycin. Background technique [0002] The resistance of pathogenic bacteria to antibiotics is one of the severe challenges facing the world today, and finding antibiotics that can effectively inhibit drug-resistant bacteria is the fundamental way to solve this problem. Vancomycin was once considered the last line of defense against Gram-positive bacteria, but more and more bacteria resistant to this drug have been found clinically all over the world. [0003] Daptomycin is the first product of a new family of antibiotics called cyclic lipopeptides. It is a novel cyclolipopeptide antibiotic composed of thirteen amino acids extracted from the fermentation broth of Streptomyces parvus, among which ten amino acids form a ring structure, and esterification occurs in the outer ring of decanoic acid and tryptophan . Not only does it have a no...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/36C07K1/34C07K1/30C07K1/22C07K1/20C07K1/18
CPCC07K7/08
Inventor 黄志强黄娟赵正国林东态蒋永飞巫秋萍
Owner LIVZON GROUP FUZHOU FUXING PHARMACEUTICAL CO LTD
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