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Prepn of active haparin

A technology of heparin and buffer, applied in the field of biopharmaceutical preparation, can solve the problems of low purity of arginine esterase, toxic substances, etc.

Inactive Publication Date: 2002-07-10
周肖林 +1
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Problems solved by technology

[0003] In order to solve the technical problem that the arginine esterase obtained by the existing method for extracting arginine esterase from Agkistrodon venom is not high in purity and contains a large amount of toxic substances, the invention provides a method for extracting and preparing from Agkistrodon halys venom in Jiangsu and Zhejiang Heparin (that is, arginine esterase) method, the purity can reach more than 90%, so as to ensure the safety of medication

Method used

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Embodiment Construction

[0012] Take 20 grams of Jiangsu-Zhejiang Agkistrodon venom freeze-dried powder, dissolve in 20ml of 0.05M Tris-HCl, pH7.8 buffer solution, centrifuge in a refrigerated centrifuge at 3000 rpm for 10 minutes, remove insoluble matter, and add the supernatant to DEAE cellulose DE-52 chromatographic column, 3×80cm, the chromatographic column needs to be equilibrated with Tris-HCl buffer first, the eluent is gradient eluted with a concentration of 0 to 0.5M containing NaCl, and the arginine esterase activity is collected Components (detected at a wavelength of 280nm by an ultraviolet spectrophotometer), the collected liquid was combined, concentrated by ultrafiltration with an ultrafiltration membrane with a molecular weight cut-off of more than 10,000, dialyzed to remove salt, and then separated by CM-Sepharose column chromatography. The size is 2×40cm; equilibrate with 0.05M sodium acetate buffer solution pH5.0, after adding the sample, carry out gradient elution with sodium acetat...

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Abstract

The present invention relates to the method of extracting liver activating essence from Jiangsu and Zhejiang pit viper venom. The liver activating essence has main component arginine esterase and has the pharmacological functions of reducing blood viscosity, improving microcirculation, inhibiting hepatitis virus infected cell and promoting the restore of liver function. The neurotoxin, hemorrhagic toxin and other harmful components in Jiangsu and Zhejiang pit viper are eliminated through three-column chromatographic plate separation and purification and the arginine esterase content is made to reach 90%. Animal test shows that the present invention has low toxicity and is safe for clinical application.

Description

Technical field [0001] The invention relates to a method for preparing biological medicines, especially a method for extracting arginine esterase from the venom of Agkistrodon halys in Jiangsu and Zhejiang as a medicine for treating liver diseases. Using this method, high-purity arginine esterase can be obtained, making it more effective It is suitable for being used in medicines for treating liver diseases. Background technique [0002] It is known that Agkistrodon halys venom contains pharmacologically active arginine esterase. Previously, a refined Agkistrodon halys antithrombotic enzyme has obtained the approval number of Liaoning Provincial Drug Administration. It is used for the treatment of heart, brain, and vascular diseases. Salvia works well. However, due to the low purity of antithrombotic enzyme from snake venom in my country, the production of this drug was disordered by the Ministry of Health. In 1994, it was rectified. In 1997, the original production license...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P1/16C12N9/00C12N9/18
Inventor 周肖林陈远聪
Owner 周肖林
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