Preparation method of single teicoplanin components

A technology of teicoplanin and teicoplanin alone, which is applied in the field of separation and purification of teicoplanin single-component preparation, can solve the problems of low reusability of separation medium, strict requirements of separation system, and small preparation volume, and achieve The effect of controllable quality, simple and reasonable preparation process, and less solvent consumption

Active Publication Date: 2012-10-10
NCPC NEW DRUG RES & DEV
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  • Abstract
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AI Technical Summary

Problems solved by technology

In this method, TA can be obtained by using silanized silica gel column and preparative high performance liquid chromatography. 2-1 ~TA 2-5 Five single components, but the separation medium used has low reusability, and preparative chromatography has strict requirements on the separation system, resulting in high cost and small preparation volume of the method

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  • Preparation method of single teicoplanin components
  • Preparation method of single teicoplanin components
  • Preparation method of single teicoplanin components

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preparation example Construction

[0050] A kind of preparation method of teicoplanin single component of the present invention, comprises the following steps:

[0051] Step 1) Pretreatment: under normal temperature conditions, add an alkaline solution to the teicoplanin fermentation broth, and adjust the pH to 11.0-13.0; preferably 11.5-12.5. The alkaline solution is sodium hydroxide solution or potassium hydroxide solution, preferably sodium hydroxide solution. The pH-adjusted fermentation broth is separated from solid and liquid by means of vacuum suction filtration, plate-and-frame filter press or centrifugation, and the obtained filtrate is adjusted to pH 7.0-7.5 with an acid solution to obtain a teicoplanin filtrate. The acid solution is one of hydrochloric acid, sulfuric acid or organic acid solution, preferably hydrochloric acid.

[0052] Step 2) Decolorization: introduce the teicoplanin filtrate into a macroporous decolorization resin for decolorization. The macroporous decolorization resin is one of ...

Embodiment 1

[0062] Take 5 L of teicoplanin fermentation broth, and the fermentation unit is 2380 μg / mL. Adjust the pH of the fermentation broth to 11.5 with sodium hydroxide solution, and vacuum filter. The filtrate was adjusted to pH 7.0 with hydrochloric acid solution, and then passed through a macroporous resin LX-98 column (loading capacity: 1200 mL) at a flow rate of 2 BV / h for decolorization. The decolorization solution is introduced into the macroporous resin HZ801 column (capacity is 1200mL) at a flow rate of 2BV / h for adsorption and enrichment. After the adsorption is completed, it is first washed with 15% methanol aqueous solution, and then desorbed with 40% methanol aqueous solution. The desorption flow rate is controlled at 0.5BV / h, when HPLC detects that teicoplanin flows out, start to collect the desorption solution. After the desorption is completed, the desorbed solution is concentrated under reduced pressure at 50° C. until the concentration of teicoplanin is 80 g / L, and...

Embodiment 2

[0064] Take 10L of teicoplanin fermentation broth, and the fermentation unit is 2700 μg / mL. Adjust the pH of the fermented broth to 12.5 with potassium hydroxide solution, and filter by centrifugation. The filtrate was adjusted to pH 7.5 with sulfuric acid solution, and then passed through a macroporous resin LSA-700B column (3000 mL capacity) at a flow rate of 2 BV / h for decolorization. The decolorization solution is introduced into the macroporous resin HZ816 column (3000mL capacity) at a flow rate of 2BV / h for adsorption and enrichment. After the adsorption is completed, it is first washed with 20% ethanol aqueous solution, and then desorbed with 50% ethanol aqueous solution. The desorption flow rate is controlled at 0.5BV / h, when HPLC detects that teicoplanin flows out, start to collect the desorption solution. After the desorption is completed, the desorbed solution is concentrated under reduced pressure at 50° C. until the concentration of teicoplanin is 100 g / L, and ac...

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Abstract

The invention relates to a preparation method of single teicoplanin components, which belongs to the field of preparation of medicaments with microbial ingredients. The method comprises the following steps of: decolorizing teicoplanin fermentation liquor through macroporous decolorizing resin, and absorbing the teicoplanin fermentation liquor through macroporous adsorption resin, then desorbing the teicoplanin fermentation liquor, and concentrating the teicoplanin fermentation liquor to obtain a crude teicoplanin extract including the six components of TA2-1, TA2-2, TA2-3, TA2-4, TA2-5 and TA3-1; and dissolving the crude extract in water, and performing chromatographic separation on the dissolved crude extract through polymer microspheres to obtain the six single components, namely TA2-1, TA2-2, TA2-3, TA2-4, TA2-5 and TA3-1, of teicoplanin with high purity. According to the separation method of the single teicoplanin components, the novel chromatography media-polymer microspheres are adopted to simultaneously separate the six single components from the crude teicoplanin extract; and the method is simple in technology, can be used for disposing a large quantity of samples, consumes less solvent, can be used for preparing the single teicoplanin components in a large scale, and provides samples for intensive study on pathology, toxicology and the like of the single teicoplanin components.

Description

technical field [0001] The invention relates to a preparation method of traditional Chinese medicine raw materials in the technical field of industrial microorganisms, in particular to a preparation method of separating and purifying a single component of teicoplanin from a fermentation culture. Background technique [0002] Teicoplanin (Teicoplanin), for the genus Actinomycetes ( Actinoplanes teichomyceticus ) is a glycopeptide antibiotic produced by fermentation. It is composed of Teicoplanin A 2 (TA 2 ) five structurally similar glycopeptides (TA 2-1 ~TA 2-5 ) and deacylglucosamine Teicoplanin A 3 (TA 3 ) mixture. Teicoplanin is another glycopeptide antibiotic against drug-resistant bacteria developed after vancomycin. It mainly has strong antibacterial activity against Gram-positive aerobic and anaerobic bacteria, especially against MRSA-induced Infection has a good curative effect, and it is one of the few clinically active drugs against multi-drug resistant Sta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K9/00C07K1/36C07K1/22C07K1/16
Inventor 张雪霞任风芝段宝玲董爱华蒋沁张志江李晓露李宁王海燕朱秀良林毅陈书红成晓迅李丽红
Owner NCPC NEW DRUG RES & DEV
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