Preparation method of hirudin crude extract

A technology of crude extract and hirudin, applied in the direction of leech inhibitors, chemical instruments and methods, medical preparations containing active ingredients, etc., can solve the problems of low preparation efficiency, difficulty in guaranteeing, high product activity, etc., and achieve improvement disease, improve production efficiency, and ensure the effect of product quality

Inactive Publication Date: 2017-02-01
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims at the technical defects of the prior art, and provides a method for preparing the crude extract of hirudin, so as to solve the technical problem that the hirudin preparation method in the prior art is difficult to ensure high product activity
[0005] Another technical problem to be solved by the present invention is that the production efficiency of hirudin preparation method in the prior art is relatively low

Method used

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  • Preparation method of hirudin crude extract
  • Preparation method of hirudin crude extract

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A preparation method of hirudin crude extract, comprising the following steps:

[0024] (1) Selection and cleaning: select 1 kg of healthy wide-bodied leeches in the mid-growth stage (sub-adult stage), wash them twice with water for injection at 10°C, starve for 18 hours, and freeze them at -20°C after packaging;

[0025] (2) Crushing: place the frozen material at -15°C to balance the temperature, use a frozen meat grinder to coarsely grind it once, add 1L of 10°C injection water and mix well;

[0026] (3) Colloid grinding and twisting twice, adding 0.5L 10°C water for injection and mixing to obtain a homogenate;

[0027] (4) The homogenate was quickly frozen at -20°C, quickly thawed at 37°C, and repeated freezing and thawing 6 times;

[0028] (5) The freeze-thaw feed solution was centrifuged at 8000r / min at 10°C to remove the precipitate, and 1.6L of the centrifugate was collected;

[0029] (6) The centrifugate is ultrafiltered by an ultrafiltration device, washed an...

Embodiment 2

[0032] A preparation method of hirudin crude extract, comprising the following steps:

[0033] (1) Selection and cleaning: select 1 kg of healthy wide-bodied leeches in the mid-growth stage (sub-adult stage), wash them three times with 8°C water for injection, starve for 34 hours, and freeze them at -20°C after packaging;

[0034] (2) Crushing: place the frozen material at -15°C to balance the temperature, use a frozen meat grinder to coarsely grind it once, add 1L of 8°C injection water and mix well;

[0035] (3) Colloid milling and twisting 3 times, adding 1L of 10°C water for injection and mixing to obtain a homogenate;

[0036] (4) The homogenate was quickly frozen at -20°C, quickly thawed at 37°C, and repeatedly frozen and thawed 8 times;

[0037] (5) Centrifuge the freeze-thaw feed solution at 15°C at 10,000 r / min to remove the precipitate, and collect 2.3 L of the centrifugate;

[0038] (6) The centrifugate is ultrafiltered by an ultrafiltration device, washed and fil...

Embodiment 3

[0041] A preparation method of hirudin crude extract, comprising the following steps:

[0042] (1) Selection and cleaning: select 1 kg of healthy wide-bodied leeches in the mid-growth stage (sub-adult stage), wash them twice with water for injection at 10°C, starve for 48 hours, and freeze them at -20°C after packaging;

[0043] (2) Crushing: place the frozen material at -15°C to balance the temperature, use a frozen meat grinder to coarsely grind it once, add 2L of 5°C injection water and mix well;

[0044] (3) Colloid grinding and twisting twice, adding 1L of 5°C water for injection and mixing to obtain a homogenate;

[0045] (4) The homogenate was quickly frozen at -20°C, quickly thawed at 37°C, and repeatedly frozen and thawed 7 times;

[0046] (5) The freeze-thaw feed solution was centrifuged at 9000r / min at 15°C to remove the precipitate, and 2.5L of the centrifugate was collected;

[0047] (6) The centrifugate is subjected to ultrafiltration through an ultrafiltration...

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Abstract

The invention provides a preparation method of a hirudin crude extract. The technical scheme is as follows: by using Whitmania pigra as the raw material, selecting, cleaning, refrigerating, crushing, precipitating, centrifugating and ultrafiltering to prepare the polypeptide compound product hirudin. The hirudin has the anticoagulation action, can be used for improving the treatment effect on neoplastic or thrombotic diseases in clinical veterinary medicine, and is especially suitable for neoplastic or thrombotic diseases of dogs, cats and other animals. The refrigerating crushing and ultrafiltration techniques are combined to extract the hirudin crude product, thereby ensuring the product quality on the premise of controlling the temperature. The method is suitable for the demands of mass production, can easily implement automatic production, and enhances the production efficiency. The prepared hirudin product lays a foundation for further purification and clinical application. The product can be used for neoplastic or thrombotic diseases of cats, dogs and other pets, effectively improves the state of illness, and provides a novel biological product for the pet market.

Description

technical field [0001] The invention belongs to the field of biotechnology, and further relates to a preparation method of veterinary drug biological products, in particular to a preparation method of hirudin crude extract. Background technique [0002] Hirudo is a representative animal medicine for promoting blood circulation and removing blood stasis. It has a long history of clinical application and can significantly improve the symptoms of patients with blood stasis syndrome. Its active ingredient, hirudin, has been successfully extracted and widely used in clinical practice. Natural hirudin is produced in leeches In the saliva of the leech, it is secreted when the leech sucks blood. Natural hirudin is a single-chain polypeptide consisting of 65 or 66 amino acid residues, with a molecular weight of 7kD. Hirudin is a direct inhibitor of thrombin, which can affect blood coagulation and promote fibrinolysis from multiple links, and can be used for anti-inflammatory Coagula...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/815C07K1/34C07K1/14A61K38/58A61P7/02A61P35/00
CPCC07K14/815A61K38/58
Inventor 王尚尚苏建东乔玲付旭彬李守军
Owner TIANJIN RINGPU BIO TECH
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