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Lumbricus kinase of electrophoretical purity and its prepn and use

A lumbrokinase and electrophoresis technology, which is applied in the field of lumbrokinase and its preparation, can solve the problems of complex separation and purification process, difficulty in industrialized expansion of production and the like, and achieves the effects of high fibrinolytic activity, easy amplification and simple preparation method.

Inactive Publication Date: 2005-09-07
BEIJING BAIAO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its separation and purification process is complicated, and it is difficult to industrialize and expand production

Method used

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  • Lumbricus kinase of electrophoretical purity and its prepn and use
  • Lumbricus kinase of electrophoretical purity and its prepn and use
  • Lumbricus kinase of electrophoretical purity and its prepn and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Take 5 grams of oral lumbrokinase raw material produced by Beijing Bio Pharmaceutical Co., Ltd., batch number 20010913-3, with a specific activity of 12090.8 units / mg, fully dissolve it with 0.02M phosphate buffer with pH 7.5 and set the volume to 100 mL, filter through a 0.45 μm microporous membrane to get the filtrate. A 20ml DEAESepharose Fast Flow anion exchange chromatography column was equilibrated with 0.02M phosphate buffer at pH 7.5, loaded with 20ml of sample, washed for one column volume, and eluted with a gradient of 0-0.5M NaCl phosphate buffer. The time is 30 minutes, and about 15 milliliters of protein peaks with fibrinolytic activity are collected. The samples obtained from 5 times DEAESepharose Fast Flow anion exchange chromatography were combined, desalted and concentrated with a 3K ultrafiltration cup, and replaced with 0.02M barbiturate monosodium chloride buffer solution with pH 7.8 to a volume of 10 ml. Use 20 milliliters of Q Sepharose HP anion c...

Embodiment 2

[0034] Get 10 grams of oral lumbrokinase crude drug produced by Beijing Baiao Pharmaceutical Co., Ltd., batch number 20010913-3, and the specific activity is 12090.8 units / mg, fully dissolved with 0.02M barbiturate-sodium chloride buffer solution of pH7.8 and The volume was adjusted to 100 ml, and the filtrate was obtained by filtration through a 0.45 μm microporous membrane. A 20ml Q Sepharose XL anion exchange chromatography column was equilibrated with 0.02M barbiturate-sodium chloride buffer at pH 7.8, loaded with 25ml of sample and washed for one column volume with 0-1M NaCl barbiturate-chloride buffer Gradient elution with sodium chloride buffer, the elution time is 50 minutes, and about 20 ml of protein peaks with fibrinolytic activity are collected. The samples obtained from the 4 times of Q Sepharose XL anion exchange chromatography column chromatography were combined, desalted and concentrated with a 3K ultrafiltration cup, and replaced with 0.02M Tris-HCl buffer sol...

Embodiment 3

[0036]Take 50 grams of oral lumbrokinase raw material produced by Beijing Baiao Pharmaceutical Co., Ltd., batch number 2002120, with a specific activity of 15507.6 units / mg, fully dissolve it with 0.02M phosphate buffer solution of pH 7.5 and make it to 1000 ml, 0.45 Filter the filtrate through a μm microporous membrane. A 300ml QSepharose XL anion exchange chromatography column was equilibrated with 0.02M phosphate buffer at pH 7.5, loaded with 500ml of sample, washed one column volume, and eluted with a gradient of 0-0.5M NaCl phosphate buffer, the elution time For 75 minutes, about 300 ml of protein peak with fibrinolytic activity was collected. The samples obtained from the two Q Sepharose XL anion-exchange chromatography column chromatography were combined and concentrated with a 5K ultrafiltration system, and replaced with 0.02M barbiturate-sodium chloride buffer solution with pH 7.8 to a volume of 200 ml. Use 300 milliliters of QSepharose HP anion-exchange chromatograp...

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Abstract

The present invention relates to one kind of lumbricus kinase of electrophoretical purity and the lumbricus kinase is extracted from Chiziaisheng lumbricus and may be used in treating ischemic cerebral vascular diseases. The lumbricus kinase as one kind of proteinase is prepared via ion exchange chromatography, ultrafiltering and gel filtering to desalt. It has electrophoretical purity and may be used as the material of freeze dried powder for injection and medicine liquid for intravenous injection. The lumbricus kinase of the present invention has low toxicity and high fibrinolytic activity, and is safe and effective.

Description

technical field [0001] The invention relates to a lumbrokinase extracted from Eisenia foelide and used for treating ischemic cerebrovascular diseases, a preparation method and application thereof. Background technique [0002] Cardiovascular and cerebrovascular diseases are major diseases that seriously endanger human health and life. The coagulation factor fibrinogen (Fg, coagulation factor I) with the highest content in plasma is an independent risk factor for cardiovascular and cerebrovascular diseases. Reducing fibrinogen is the main clinical method for treating thrombotic diseases such as cerebral infarction, so the development of drugs for reducing fibrinogen has significant economic and social benefits. [0003] At present, there are many drugs used clinically for the treatment of ischemic cerebrovascular diseases by injection, including streptokinase, urokinase, batroxobin, tissue plasminogen activator and so on. Some of these drugs have strong antigenicity, no obvi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/48A61P7/02A61P9/10C12N9/48C12N9/64
Inventor 刘建蓉梅运红李树东侯全民
Owner BEIJING BAIAO PHARMA
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