TAT protein transduction modified NEP1-40 fusion protein and use thereof

A technology of protein transduction domain and fusion protein, which is applied in the fields of application, peptide/protein components, medical preparations containing active ingredients, etc., can solve problems affecting drug research and development, achieve high transduction efficiency and overcome limitations Effect

Inactive Publication Date: 2009-04-29
FOURTH MILITARY MEDICAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

The main regulatory substances of cells and intercellular physiological reactions in the central nervous system are peptides and proteins, so peptide or protein macromolecular drugs are of great significance in the treatment of CNS diseases, but because peptide or protein macromolecular drugs are difficult to Enter brain tissue and cells through the BBB, thus affecting the research and development of such drugs

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  • TAT protein transduction modified NEP1-40 fusion protein and use thereof
  • TAT protein transduction modified NEP1-40 fusion protein and use thereof
  • TAT protein transduction modified NEP1-40 fusion protein and use thereof

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Embodiment Construction

[0020] Below in conjunction with specific example, further set forth the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific condition in the following examples, generally according to conventional conditions such as people such as Sambrook, molecular cloning: the condition described in the laboratory handbook (New York: Cold Spring Harbor Laboratory Press, 1989); Another example such as David Human, conditions described in Cellular Experiment Manual (New York: Cold Spring Harbor Laboratory Press, 1998).

[0021] (1) Expression, purification and renaturation of TAT-NEP1-40 fusion protein

[0022] 1. Preparation of Escherichia coli competent cells (CaCl 2 Law):

[0023] (1) Pick a single colony of Escherichia coli DE3 in 5 ml of LB medium without Amp, and culture at 37°C overnight.

[0024] (2)...

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Abstract

The invention discloses a preparation method for TAT-NEP1-40 fusion protein and application in treating the injuries of a central nervous system, CNS. The TAT-NEP1-40 fusion protein is characterized in that the TAT-NEP1-40 fusion protein comprises the protein transduction domain (PTD) of a transactivating transduction protein TAT of the human immunodeficiency virus (HIV) and Nogo extracellular peptide residues 1-40, NEP1-40. The fusion protein retains the selective antagonistic activity of the NEP1-40 to a Nogo receptor, NgR, and has the advantages of high transduction efficiency and easy penetration of BBB and BCSB, thereby overcoming the limitation of the NEP1-40, avoiding the extra injuries on nervous tissues caused by the mode of injecting the NEP1-40 through microinjection or miniature pump implantation, and overcoming the problem that the NEP1-40 can hardly pass BBB and BCSB to reach corresponding biological concentration. The fusion protein can be prepared in a large quantity, has a low cost and high activity, and can be used for treating a plurality of CNS injuries including cerebral ischemia, cerebral anoxia, cerebral hemorrhage, cerebral trauma, spinal core injury, and the like, and promoting the regeneration and functional rehabilitation of nervous tissues.

Description

technical field [0001] The invention belongs to the field of protein transduction, and in particular relates to preparing TAT-NEP1-40 fusion protein by fusing TAT and NEP1-40. At the same time, the invention also relates to the TAT-NEP1-40 fusion protein passing through the cell membrane and the blood-brain barrier for the treatment of central nervous system damage. Background technique [0002] Trauma, stroke, cerebral hemorrhage, and spinal cord injury are characterized by high mortality and disability. How to reduce the degree of neurological damage and how to better improve the function of damaged neurons and improve the quality of life of patients is of great significance to patients, their families and society, and is a worldwide problem. Currently, there is no effective treatment for neurological dysfunction caused by CNS injury. Therefore, finding new and effective drugs for the treatment of brain CNS injury is a research hotspot. [0003] In the past, it was gene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/49C12N15/12C12N15/63A61K38/16A61P25/00
Inventor 王强熊利泽苟兴春陈绍洋
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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