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Preparation of effective expressed human DNA enzyme repression factors and their uses in medicines against oxidation and its use against generation of blood vessels

A ribonuclease and inhibitor technology, applied in the biological field, can solve problems such as short half-life, and achieve the effect of inhibiting the formation of abnormal tissue blood vessels and inhibiting the formation of tumors

Inactive Publication Date: 2005-02-23
崔秀云 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the short half-life of the inhibitory factors of neovascularization, daily administration is required, and the dose usually required is also relatively large, such as endostatin (endostatin) and angiopoietin (angiostatin) at a therapeutic dose of 10-20 mg / kg / day

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Application of human ribonuclease inhibitor in the preparation of anti-tumor angiogenesis drugs.

[0027]RI preparations were extracted from prokaryotic (E. coli) and eukaryotic cells transformed with the RI gene. The process flow is as follows: After collecting the amplified and cultured Escherichia coli or yeast, the cells are disrupted by ultrasonic waves, proteins are separated, and RI is purified by affinity chromatography. Determination of the content of RI and its activity (inhibition of RNase A activity). Freeze-dried, subpackaged and stored to make RI preparations.

[0028] Experimental studies of the use of human ribonuclease inhibitors in anti-tumor angiogenesis drugs. Inhibitory effect of ribonuclease inhibitor on the growth of S-180 sarcoma in mice

[0029] The RI purified by affinity chromatography was injected subcutaneously into pure line L-615 tumor-bearing mice (10000U / kg). Administration was given once a day for 5 consecutive days. The control gro...

Embodiment 2

[0031] Application of human ribonuclease inhibitor in the preparation of anti-angiogenic drugs.

[0032] RI preparations were extracted from prokaryotic E. coli and eukaryotic cells transformed with the RI gene. The process flow is as follows: after the amplified cultured Escherichia coli or yeast are collected, the cells are disrupted by biochemical techniques, proteins are separated, and RI is purified by affinity chromatography. Determination of the content of RI and its activity (inhibition of RNase A activity). Freeze-dried, compressed into tablets, packaged and preserved.

[0033] Under normal conditions, blood vessels in the mammalian eye stop growing, partly because of the presence of angiostatic factors, which prevent blood vessels from eroding into the cornea and lens (vitreous). The growth of new blood vessels is tightly regulated in most healthy tissues, and when this suppression is out of balance, it causes many diseases, from joints to tumors. In retinal ische...

Embodiment 3

[0038] Application of human ribonuclease inhibitor in the preparation of antioxidant drugs.

[0039] RI preparations were extracted from prokaryotic (E. coli) and eukaryotic cells transformed with the RI gene. The process flow is as follows: after the amplified cultured Escherichia coli or yeast are collected, the cells are broken by biochemical techniques, proteins are separated, and RI is purified by ion exchange chromatography. Determination of the content of RI and its activity (inhibition of RNase A activity). Pack and save.

[0040] RI is a protein rich in cysteine, which contains 30 free sulfhydryl groups in the RI molecule. According to its molecular structure characteristics, our research has confirmed that the RI molecule has an antioxidant effect.

[0041] The experimental results are as follows:

[0042] a. Using the gene transfection method, the RI gene was transferred into the glioma C6 cell line (C'6). with different concentrations of H 2 o 2 cause C6 cell...

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Abstract

An efficient expressing human ribonuclease inhibiting factor preparing method, use for antivessel formation and antioxidant medicine are disclosed. It relates to efficient expression of recombinant ribonuclease inhibiting factor in prokaryotic and eucaryotic cell, recombinant ribonuclease inhibiting factor for inhibiting tumor vessel formation, formation and transition of tumor. It can be used for treating glaucoma and cataract, antioxidation and antisenescence.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to the high expression of recombinant ribonuclease inhibitors in prokaryotic and eukaryotic cells and the application of highly expressed ribonuclease inhibitors in anti-angiogenesis and antioxidant drugs. Background technique [0002] Ribonuclease Inhibitor (Ribonuclease Inhibitor, RI) is a protein that can bind tightly to angiogenesis factor (angiogenin) and inhibit its activity. During the growth of malignant solid tumors, the formation of new blood vessels is of great significance. The growth and metastasis of tumors must rely on continuous and extensive angiogenesis. It can not only provide nutrients, oxygen and necessary growth factors for tumor tissues, but also excrete metabolites. Once there are new blood vessels, the tumor grows rapidly and metastasizes. Metastasis is the greatest threat to cancer patients. The whole process from the occurrence of metastasis to the final format...

Claims

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Application Information

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IPC IPC(8): A61K38/55A61P39/06C12N15/15C12N15/79C12P21/00
Inventor 崔秀云赵宝昌
Owner 崔秀云
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