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Non-viral gene vector for gene transfer as well as preparation method and use thereof

A gene vector, non-viral technology, applied to the non-viral gene vector for gene transfer and the fields of its preparation and use, can solve the problems of unsatisfactory transfection efficiency, reduced cytotoxicity and the like

Active Publication Date: 2017-05-31
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the cytotoxicity of PEI-anionic polymer complexes is greatly reduced, the transfection efficiency is always unsatisfactory

Method used

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  • Non-viral gene vector for gene transfer as well as preparation method and use thereof
  • Non-viral gene vector for gene transfer as well as preparation method and use thereof
  • Non-viral gene vector for gene transfer as well as preparation method and use thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 The present invention is used for the preparation of the non-viral gene carrier (HPR) of gene delivery

[0029] 1. Preparation method

[0030] (1) Preparation of PEI-R8

[0031] Dissolve PEI 1.8K (1 g, 0.55 mmol, 1.0 eq) in 10 mL of chloroform in a 25 mL round bottom flask, then add 3-maleimido propionate hydroxysuccinimide ester (177 mg, 0.666 mmol, 1.2 eq). The solvent was removed after the reaction was stirred at room temperature for 3 hours. Then with 10mL THF / H 2 O (1:1, v / v) dissolved the above residue, then added cys-R8 (760mg, 0.55mmol, 1.0eq) and stirred at room temperature for 6 hours. After the reaction was complete, the above reaction solution was dialyzed in water for three days with a dialysis bag with a molecular weight cut off of 1000. Finally, the aqueous solution of the obtained product was lyophilized and stored at -20°C for future use.

[0032] (2) Preparation of HPR

[0033] Dissolve 50 mg of heparin sodium in 100 mL of MES buffe...

Embodiment 2

[0040] Embodiment 2 The present invention is used for the preparation of the non-viral gene carrier (HPR) of gene delivery

[0041] 1. Preparation method

[0042] (1) Preparation of PEI-R8

[0043] Dissolve PEI 1.8K (1 g, 0.55 mmol, 1.0 eq) in 10 mL of chloroform in a 25 mL round bottom flask, then add 3-maleimido propionate hydroxysuccinimide ester (177 mg, 0.666 mmol, 1.2 eq). The solvent was removed after the reaction was stirred at room temperature for 3 hours. Then with 10mL THF / H 2 O (1:1, v / v) dissolved the above residue, then added cys-R8 (760mg, 0.55mmol, 1.0eq) and stirred at room temperature for 6 hours. After the reaction was complete, the above reaction solution was dialyzed in water for three days with a dialysis bag with a molecular weight cut off of 1000. Finally, the aqueous solution of the obtained product was lyophilized and stored at minus 20 degrees for future use.

[0044] (2) Preparation of HPR

[0045] Dissolve 50 mg of heparin sodium in 100 mL o...

Embodiment 3

[0046] Embodiment 3 The present invention is used for the preparation of the non-viral gene carrier (HPR) of gene delivery

[0047] 1. Preparation method

[0048] (1) Preparation of PEI-R8

[0049] Dissolve PEI 1.8K (1 g, 0.55 mmol, 1.0 eq) in 10 mL of chloroform in a 25 mL round bottom flask, then add 3-maleimido propionate hydroxysuccinimide ester (177 mg, 0.666 mmol, 1.2 eq). The solvent was removed after the reaction was stirred at room temperature for 3 hours. Then with 10mL THF / H 2 O (1:1, v / v) dissolved the above residue, then added cys-R8 (760mg, 0.55mmol, 1.0eq) and stirred at room temperature for 6 hours. After the reaction was complete, the above reaction solution was dialyzed in water for three days with a dialysis bag with a molecular weight cut off of 1000. Finally, the aqueous solution of the obtained product was lyophilized and stored at minus 20 degrees for future use.

[0050] (2) Preparation of HPR

[0051] Dissolve 50 mg of heparin sodium in 100 mL o...

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Abstract

The invention discloses a non-viral gene vector for gene transfer. The non-viral gene vector for gene transfer is formed by connecting polyethyleneimine, heparin sodium and cell permeable peptides. The invention also provides a preparation method and use of the vector. The non-viral gene vector HPR is low in toxicity, high in transfection efficiency and good in clinical application prospect.

Description

technical field [0001] The invention relates to a non-viral gene vector for gene delivery and its preparation method and application. Background technique [0002] Gene therapy has attracted more and more people's attention. It can provide an efficient and potential treatment method for many diseases such as AIDS and cancer by delivering gene medicine to target cells or tissues. As we all know, since naked DNA is easily degraded and cleared by nucleases in blood or body fluids after entering the body, naked DNA is not suitable for direct use in in vivo treatment. Therefore, we urgently need a gene carrier that can not only protect the gene drug from being degraded, but also efficiently deliver the gene drug to the target cells or tissues. Although some viral vectors have high transfection efficiency, the inherent immunogenicity and biological safety of viral vectors limit their further clinical application. Recently, some non-viral gene carriers have attracted the attentio...

Claims

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Application Information

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IPC IPC(8): C12N15/63C08G73/02A61K48/00
CPCA61K48/0041C08G73/02C12N15/63
Inventor 巩长旸吴秦洁宋林江魏于全
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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