Dried lumbrukinase powder preparation method

A technology of lumbrokinase dry powder and earthworm, which is applied in the directions of biochemical equipment and methods, enzymes, hydrolase, etc., can solve the problems of low quality standard of lumbrokinase, frequent regeneration, high pressure on environmental protection, etc. Scale, the effect of improving work capacity

Active Publication Date: 2013-12-18
BEIJING BAIAO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the production of lumbrokinase in China has the characteristics of simple and extensive, low degree of automation, high pressure on environmental protection, and relatively low quality standards of lumbrokinase.
These characteristics limit the large-scale production of lumbrokinase, which is not conducive to the improvement of the quality of lumbrokinase products, and cannot well meet the needs of my country's major disease prevention and development of the pharmaceutical industry.
[0003] For example, the patent (ZL03146692.3) discloses the use of separation methods including anion exchange chromatography, ultrafiltration, gel filtration and desalination to obtain electrophoretic-grade protease, which has high production costs and complicated quality control.
The patent (ZL200410039186.8) discloses a method to increase the content of effective components of lumbrokinase, but the total activity is not high and the yield varies greatly
Due to the lack of effective pretreatment methods in the above methods, the purity of the intermediates before chromatography is low, the chromatography load is low, the medium is seriously polluted, the regeneration is frequent, and the production scale is small, the cost is high, and the total activity yield of the product is low.

Method used

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  • Dried lumbrukinase powder preparation method
  • Dried lumbrukinase powder preparation method
  • Dried lumbrukinase powder preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1) Cleaning and thawing of earthworms: wash fresh earthworms with water, wash away sludge and sticky matter, and then store them in a cold storage for later use; place 800kg of frozen earthworms at room temperature to thaw naturally;

[0035] 2) Pulp making: add water 1.0 times the volume of the earthworms in the homogenization tank, pour the completely thawed earthworms, crush the earthworms with a homogenizer, and homogenize for 2.0 hours to form a homogenate;

[0036] 3) Biological enzyme treatment: add serrapeptase in an amount of 0.1% of the weight of earthworms to the homogenate in step 2), control the temperature at 35°C, and keep the temperature for 6 hours;

[0037] 4) Centrifugation: the homogenate in step 3) is quantitatively diluted with water, and the final volume is set to 2.5 tons, and the coarse slag is removed by a butterfly centrifuge;

[0038] 5) clarification: after step 4) the centrifugate after slag removal is input into the membrane filtration sys...

Embodiment 2

[0047]1) Cleaning and thawing of earthworms: wash fresh earthworms with water, wash away sludge and sticky matter, and then store them in a cold storage for later use; place 800kg of frozen earthworms at room temperature to thaw naturally;

[0048] 2) Pulp making: add water 1.0 times the volume of the earthworms in the homogenization tank, pour the completely thawed earthworms, crush the earthworms with a homogenizer, and homogenize for 2.0 hours to form a homogenate;

[0049] 3) Biological enzyme treatment: in the homogenate in step 2), add bromelain in an amount of 0.1% of the weight of the earthworm, control it at 35° C., and keep it for 6 hours;

[0050] 4) Centrifugation: the homogenate in step 3) is quantitatively diluted with water, and the final volume is set to 2.5 tons, and the coarse slag is removed by a butterfly centrifuge;

[0051] 5) clarification: after step 4) the centrifugate after slag removal is input into the membrane filtration system, add water to wash, ...

Embodiment 3

[0060] 1) Cleaning and thawing of earthworms: wash fresh earthworms with water, wash away sludge and sticky matter, and then store them in a cold storage for later use; place 800kg of frozen earthworms at room temperature to thaw naturally;

[0061] 2) Pulp making: add water 1.0 times the volume of the earthworms in the homogenization tank, pour the completely thawed earthworms, crush the earthworms with a homogenizer, and homogenize for 2.0 hours to form a homogenate;

[0062] 3) Biological enzyme treatment: in the homogenate in step 2), add papain in an amount of 0.1% of the weight of earthworms, control at 35° C., and keep the temperature for 6 hours;

[0063] 4) Centrifugation: the homogenate in step 3) is quantitatively diluted with water, and the final volume is set to 2.5 tons, and the coarse slag is removed by a butterfly centrifuge;

[0064] 5) clarification: after step 4) the centrifugate after slag removal is input into the membrane filtration system, add water to w...

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Abstract

The invention relates to a dried lumbrukinase powder preparation method. The dried lumbrukinase powder preparation method includes the following steps of unfreezing frozen fresh earthworms at room temperature, pulping, treating by biological enzyme, centrifuging, clarifying by a filter film, removing micromolecule impurity proteins by ultrafiltration, clarifying by the filter film secondarily, adjusting the pH (potential of hydrogen) value, exchanging and layering by negative ions, obtaining clarified liquid to subject to concentration and ultrafiltration, clarifying and filtering, freezing and drying to obtain dried lumbrukinase powder. The dried lumbrukinase powder preparation method has the advantages that quality controllability and stability are high, targets are collected sectionally controllably, general active yield of lumbrukinase is increased, yield is stable, preparation cost is reduced, pollution of layering media is reduced, and large-scale industrial production is easy to realize and the like.

Description

technical field [0001] The invention relates to a preparation method of lumbrokinase dry powder, which is a method for large-scale production of lumbrokinase dry powder by using biological enzymolysis technology, and belongs to the field of biopharmaceuticals. Background technique [0002] Lumbrokinase powder is a bio-extracted medicine made from fresh earthworms, which is light yellow to yellowish brown powder. This drug is a group of proteolytic enzymes, including plasmin and plasminogen activator components, which can reduce the excessively high fibrinogen and platelet aggregation rate, improve symptoms and prevent the development of the disease. It is suitable for ischemia Treatment and prevention of cerebrovascular disease. Clinical trials have proved that the capsules of this drug have a remarkable recovery effect on ischemic cerebrovascular disease, stroke paralyzed limbs and language barriers, and are a safe oral fibrinolytic drug. After treatment, the concentratio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/68C12N9/64
Inventor 闫晓玲郭燕青张波吕学富王雪侯全民
Owner BEIJING BAIAO PHARMA
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