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Giant web bombinator proteinase inhibitor and its prepn. and pharmaceutical application

A technology of protease inhibitor, Bombina, applied in biomedicine

Inactive Publication Date: 2004-03-17
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The application of amphibian drugs has a long history in my country, but the research on their active ingredients and pharmacological properties mainly focuses on small organic molecules such as alkaloids, and there are few researches on their skin active protein peptides

Method used

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  • Giant web bombinator proteinase inhibitor and its prepn. and pharmaceutical application
  • Giant web bombinator proteinase inhibitor and its prepn. and pharmaceutical application
  • Giant web bombinator proteinase inhibitor and its prepn. and pharmaceutical application

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Experimental program
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Embodiment

[0030] 1. Preparation of Bombina grandis protease inhibitor: Bombina grandis, harvested from the mountainous area of ​​Yunnan Province, the living body was cleaned with water, put to death, peeled, the skin was cut into small pieces, soaked in normal saline, homogenized, centrifuged (10000rpm, 20 minutes), freeze-dried, and stored at low temperature.

[0031] The first step, DEAE Sephadex A-50 ion exchange: According to the above method, obtain the freeze-dried powder of the skin homogenate of the raw material Bombina grandis, and dissolve the freeze-dried powder in 0.05M Tris-HCl, containing 10mM EDTA, pH7.3 buffer, Packed in a 10KDa dialysis bag, dialyzed in the same buffer for 12 hours; DEAE SephadexA-50 (Pharmacia product) anion exchange column (500mm long, 26mm diameter) equilibrated with 0.05M Tris-HCl, containing 10mM EDTA, pH7.3 buffer, The dialyzed sample was put on the column, washed with two column volumes of the same buffer until the breakthrough peak was completel...

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Abstract

The present invnetion invention relates to large puling toad protease inhibitor, it is a single chain serine protease inhibitor, obtained from said toad skin, its apparent molecular weight under reduced condition is 67KDa, and under non reduced condition is 45KDa, isoelectric point is 4.8, saccharide content is 17-19%, has no enzyme activity, its N-terminal 31 amino acid sequence structure is: DSETHIRFLGEVYKKVDTIDFRGLVLITRAQ. Its preparation method is: toad skin is homogenized in physiological saline, centrifuge to remove sediment, collect supernatant, freeze dried, ion exchange, gel filtration and purification to obtain product. It possesses strong serine protease inhibiting activity and antitumour activity, can be used in preparation of medicine for treating tumour, gastricit, pancreatitis etc.

Description

Technical field: [0001] The invention provides a Bombina grandis protease inhibitor, a preparation method thereof and an application in pharmacy, which belong to the field of biomedicine. Background technique: [0002] Serine protease inhibitors (serine protease inhibitors, serpins) are a class of single-chain proteins, and their most basic function is to prevent proteolysis and regulate the hydrolysis balance of serine proteases. Through the regulation of serine proteases, serine protease inhibitors have an important impact on many important biochemical reactions and physiological functions in living organisms. The following 14 classes of biochemical reactions and physiological functions are regulated by serine protease inhibitors: blood coagulation, complement formation, fibrinolysis, protein folding, cell migration, cell differentiation, cell matrix remodeling, hormone formation, hormone transport, intracellular proteolysis, blood pressure Regulation, tumor suppression, ...

Claims

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Application Information

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IPC IPC(8): A61K38/57A61P1/04A61P1/18C07K1/14C07K14/435
Inventor 张云赖仞郑永唐唐绍宗李文辉
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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