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Cobratide extraction method, cobratide extracted thereby and formulation containing cobratide

An extraction method and technology of cobotide, applied in the field of cobotide extraction, can solve the problems of undisclosed formula and preparation method of cobotide enteric-coated capsules, achieve good analgesic effect, good bioavailability, and simple method Effect

Active Publication Date: 2009-03-11
BEIJING SAISHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the specific formula and preparation method of Kebotide enteric-coated capsules are not disclosed in this article.

Method used

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  • Cobratide extraction method, cobratide extracted thereby and formulation containing cobratide
  • Cobratide extraction method, cobratide extracted thereby and formulation containing cobratide
  • Cobratide extraction method, cobratide extracted thereby and formulation containing cobratide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] 1) Dissolve 20 g of Hunan Cobra Venom Powder (purchased from Hunan Yiyang Snake Farm) in 500 mL of 0.02M pH 6.0 phosphate buffer, centrifuge at 4000 rpm for 10-20 min, take the supernatant, add ammonium sulfate to a saturation of 60 %, precipitate overnight at 4°C, centrifuge at 4000rpm for 15-20min, discard the precipitate, and obtain the supernatant.

[0051] 2) Take the obtained supernatant and use a hollow fiber ultrafilter with a molecular weight cut-off of 6K (6K, Beijing Xubang Membrane Ultrafiltration Equipment Co., Ltd.) to desalinate and concentrate.

[0052] 3) A CM cation exchange column (column material: CMSpharose™ Fast Flow, Amersham Pharmacia BiothchAB) was equilibrated with 0.02M phosphate with a pH value of 6.0. After the concentrated solution was loaded, it was eluted with a linear gradient of 0-0.6M NaCl.

[0053] 4) Take the peak with Cobotide active ingredient, desalinate and concentrate as in step 2), and then freeze-dry to obtain Cobotide sample ...

Embodiment 2

[0065] In step 1), add ammonium sulfate to 40% of saturation, others are the same as embodiment 1. The yield is 67.3%, the molecular weight is about 7000 Daltons, and the sample purity is 98.9%.

Embodiment 3

[0067] In step 1), add ammonium sulfate to 80% of saturation, others are the same as embodiment 1. The yield is 73.4%, the molecular weight is about 7000 Daltons, and the sample purity is 97.2%.

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Abstract

The invention relates to a method for extracting cobratide, a medicine used to treat chronic pain, the cobratide extracted by the method and a preparation containing the cobratide. The method for extracting the cobratide comprises the following steps: firstly, a stock solution of snake poison of a cobra is pretreated with ammonium sulphate to remove partial other compositions in the snake poison,is subjected to desalination and concentration through ultrafiltration, dialysis or gel chromatography, and after the concentration, is purified by a cation column, and finally is subject to freeze-drying preservation after the concentration. The cobratide preparation comprises an enteric preparation, a rectal administration preparation, a colon-specific administration preparation and so on. Withcobratide extraction and purification processes provided by the method, the purity of the extracted cobratide is more than 95 percent, and the yield reaches 70 percent; and the method is simple and reliable, and saves cost and time.

Description

technical field [0001] The invention relates to a method for extracting kobotide, kobotide extracted by the method and a preparation containing the kobotide. Background technique [0002] Cobotide is a snake venom neurotoxin extracted from cobra snake venom. Snake venom neurotoxins can be divided into two types according to different physiological activities: presynaptic neurotoxins and postsynaptic neurotoxins. Cobotide belongs to the post-synaptic neurotoxin, which is a small single-chain protein composed of 68 amino acids. It does not contain non-protein components such as sugar, contains four disulfide bonds, and has a molecular weight of about 7000 Daltons. [0003] Cobra venom neurotoxin has a high affinity with N-type acetylcholine receptors, which can prevent the transmission of nerve impulse signals at neuromuscular junctions. It affects the metabolism of acetylcholine in the brain of experimental animals, and can increase the content of enkephalin in the brain of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/46A61K38/17A61K9/00A61P25/04
Inventor 马骉孔双泉宋梦薇宋法宝戴杰森孙爽
Owner BEIJING SAISHENG PHARMA
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