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Application of super oxygen dehydrogenises-trans-activator transcription (SOD-TAT) fusion protein to radiation injury prevention and cure medicine preparation

A 1. SOD-TAT, fusion protein technology, applied in the field of fusion protein, can solve the problems of inability to cure radiation damage, failure to achieve radiation protection effect, combination and other problems

Inactive Publication Date: 2014-03-26
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the selective permeability of tissues and cells, the drug cannot pass through the cell membrane to reach the site of action in the cell and combine with the target molecule, thus losing its curative effect
[0008] Since free radicals are produced by the ionizing radiation of water molecules and the respiratory burst of macrophages, they not only exist outside the cells, but also in a large amount inside the cells. Only removing free radicals outside the cells naturally cannot cure radiation damage
Therefore, although SOD is a well-known free radical scavenger, it is strictly limited by its size and other characteristics and cannot penetrate the cell membrane. However, no matter how much it is modified and how much it is used, its radiation protection effect will not meet people's expectations.

Method used

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  • Application of super oxygen dehydrogenises-trans-activator transcription (SOD-TAT) fusion protein to radiation injury prevention and cure medicine preparation
  • Application of super oxygen dehydrogenises-trans-activator transcription (SOD-TAT) fusion protein to radiation injury prevention and cure medicine preparation
  • Application of super oxygen dehydrogenises-trans-activator transcription (SOD-TAT) fusion protein to radiation injury prevention and cure medicine preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Radioprotective effect on normal cells

[0029] (1) Protective effect of SOD-TAT on the clonogenic ability of L-02 cells

[0030] 1. Cell lines

[0031] Human normal liver cell line L-02 was purchased from Shanghai Institute of Cells, Chinese Academy of Sciences.

[0032] 2. Operation

[0033]Take cells in the logarithmic growth phase, digest, prepare cell suspension, and disperse the cells sufficiently so that the percentage of single cells is greater than 90%; after accurate counting, mix well with cell culture medium containing 10% serum, and distribute to culture medium with the same specification The normal group was not administered and irradiated, and the other groups were irradiated with 2 Gy X-rays. The radiation control group was not administered before irradiation. high-efficiency expression, and finally extracted high-purity SOD-TAT fusion protein from the fermentation broth of Pichia pastoris by ion exchange) SOD-TAT was added 3 h before irr...

Embodiment 2

[0038] Example 2: Protective effect on radiation-induced lung injury in thorax-irradiated mice

[0039] 1 experimental animal

[0040] Male C57BL / 6 mice were bred by Shanghai Slack Experimental Animal Co., Ltd., clean grade. The weight is 18-21 g. Drinking water and eating freely, the feed is special irradiated material. Exhaust regularly. There was no abnormality in feeding for three days before irradiation. The experimental animal quality certificate number is SCXK (Shanghai) 2007-0005, and the animal experiment condition certificate number is SYXK (Army) 2007-031.

[0041] 2. Irradiation conditions and experimental grouping

[0042] The C57BL / 6 mice used in the experiment were randomly divided into 4 groups: normal control group (no administration and no radiation), radiation control group (only radiation without administration), amifostine group (150 mg / kg) (before radiation) 30 min injection), SOD-TAT group (6000 U / ml) (injection 3 h before radiation). X-ray 6 Me...

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Abstract

The invention relates to the field of fusion protein, in particular to an application of super oxygen dehydrogenises-trans-activator transcription (SOD-TAT) fusion protein to radiation injury prevention and cure medicine preparation. A preparation method of the SOD-TAT fusion protein has the steps that: the genetic recombination technology is adopted, TAT is fused to the C end of SOD according to gene sequences of human CuZn super oxygen dehydrogenises in a genbank, a new protein SOD-TAT is formed, expression plasmids fused with the SOD and the TAT are built, the efficient expression in pichia pastoris is obtained, and finally, high-purity SOD-TAT fusion protein is extracted from fermentation liquor of the pichia pastoris through ion exchange; and the SOD-TAT fusion protein is used for preparing radiation injury prevention and cure medicine. The SOD-TAT can eliminate free radicals outside cells and can also eliminate free radicals in the cells, in addition, the SOD-TAT can also overcome hemato encephalic barrier, and the depth and the range of the radiological protection effect are far higher than those of radiation protection agents clinically used in the prior art.

Description

technical field [0001] The invention relates to the field of fusion proteins, and more specifically relates to the application of a SOD-TAT fusion protein in the preparation of medicines for preventing and treating radiation damage. Background technique [0002] Malignant tumors are currently one of the major diseases that seriously affect human health and threaten human life. Together with cardiovascular and cerebrovascular diseases and accidents, cancer constitutes the three major causes of death in all countries in the world today. According to the statistics of the World Health Organization (WHO) and the American Society of Clinical Oncology (ASCO), more than 10 million people worldwide suffer from cancer every year, and about 7.6 million people die of various types of cancer. [0003] Radiation therapy is an effective means of killing tumors, and it undertakes the treatment of 2 / 3 tumor patients. However, the side effects of radiation damage caused by free radicals gen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/44A61P39/06A61P17/16C12N15/81C12R1/84
Inventor 潘剑茹刘树滔饶平凡郑光进
Owner FUZHOU UNIV
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