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High purity venom kininogenase prepartion method and its pharmaceutical formulation

A kininogenase and high-purity technology, applied in the field of high-purity snake venom kininogenase, can solve the problems of application scope, indications and its curative effect limitations, low purity, etc., and achieve precious treatment time, high purity, and reduce wastage. Effect

Active Publication Date: 2008-06-18
BEIJING SAISHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the snake venom kininogenase preparation, due to its low purity, can only be used for oral and intramuscular injection due to safety restrictions, which greatly limits its application range, indications and curative effect.

Method used

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  • High purity venom kininogenase prepartion method and its pharmaceutical formulation
  • High purity venom kininogenase prepartion method and its pharmaceutical formulation
  • High purity venom kininogenase prepartion method and its pharmaceutical formulation

Examples

Experimental program
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Effect test

Embodiment 1

[0057] [Example 1] Preparation of immune antibody affinity chromatography column

[0058] Purified and characterized by high performance liquid chromatography, 50μg / ml of snake venom kininogenase (active component 2) was mixed and emulsified with an equal volume of complete adjuvant as a stimulus. The 7-week-old BALB / c mice were subcutaneously injected and immunized. 0.2ml each time, once a week, a total of 6 immunizations. Three days before serum collection, 0.2ml of stimulant mice were injected into the tail vein for booster immunization. The antibody was purified using an antigen affinity chromatography column.

[0059] The 4B-Sepharose agarose gel was activated with cyanogen bromide, and the obtained snake venom kininogenase antibody was coupled to a polysaccharide matrix to form an immunoaffinity chromatography column.

Embodiment 2

[0060] [Example 2] Preparation of immunoantibody affinity chromatography column

[0061] Purified and characterized by high performance liquid chromatography, 250μg / ml of snake venom kininogenase (active component 2) was mixed and emulsified with an equal volume of complete adjuvant as a stimulant. The 7-week-old BALB / c mice were subcutaneously injected and immunized. 0.2ml each time, once a week, a total of 6 immunizations. Three days before serum collection, 0.2ml of stimulant mice were injected into the tail vein for booster immunization. Snake venom kininogenase antibody was purified by an antibody affinity chromatography column.

[0062] The diethylaminodextran gel is activated with cyanogen bromide, and the obtained snake venom kininogenase antibody is coupled to a polysaccharide matrix to form an immunoaffinity chromatography column.

Embodiment 3

[0063] [Example 3] Preparation of immune antibody affinity chromatography column

[0064] Using high performance liquid chromatography purified and qualitative snake venom kininogenase (active component 2) 500μg / ml mixed with an equal volume of complete adjuvant to emulsify as a stimulus, subcutaneous injection of 7-week-old BALB / c mice, 0.2ml each time, once a week, a total of 6 immunizations. Three days before serum collection, 0.2ml of stimulant mice were injected into the tail vein for booster immunization. Snake venom kininogenase antibody was purified by an antibody affinity chromatography column.

[0065] The carboxymethyl dextran gel is activated with cyanogen bromide, and the obtained snake venom kininogenase antibody is coupled to a polysaccharide matrix to prepare an immunoaffinity chromatography column.

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Abstract

The invention relates to a process for preparing high purity snake venom kininogenase, which comprises integrating antigen-antibody reaction with compatible chromatography technology, and isolating and purifying kininogenase from raw material snake venom. The invention also relates to the pharmaceutical preparation from the high purity snake venom kininogenase.

Description

Technical field [0001] The invention relates to a method for preparing high-purity snake venom kininogenase, and specifically relates to a high-purity snake venom kininogenase prepared by immunoaffinity chromatography. [0002] The invention also relates to the high-purity snake venom kininogenase and its preparation prepared by the method of the invention. Background technique [0003] Kininogenase is a proteolytic enzyme in animals, which exists in the tissues of many organs. It is the pancreatic kininogenase in snake venom that is the earliest known and most clearly studied. Pancreatic Kallidinogenase (Pancreatic Kallidinogenase), also known as Pancreatic Kallikrein, is a type of endoproteolytic enzyme, which exists in a variety of tissues in mammals, especially snake venom. Acts on kininogen in the body to release kinin, thereby exerting a series of pharmacological effects: dilate capillaries and improve peripheral blood circulation; promote prostaglandin secretion, expand ar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00C07K16/40A61K38/43
Inventor 马骉魏化伟吴丹
Owner BEIJING SAISHENG PHARMA