Arginine modified polyethyleneimine as well as preparation method and application thereof

A technology of polyethyleneimine and arginine, which is applied in the field of chemical medicine, can solve the problems of high toxicity, PEI application limitations, and inability to meet gene therapy, etc., to achieve improved transfection efficiency, low cytotoxicity, and high gene transfection efficiency Effect

Active Publication Date: 2021-01-12
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High-molecular-weight PEI (>25000) has a certain transfection efficiency, but is more toxic; low-molecular-weight PEI (<10000) has low toxicity, but no transfection efficiency, which cannot meet the needs of gene therapy
The mutual restriction between PEI transfection efficiency and toxicity leads to certain restrictions on the application of PEI

Method used

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  • Arginine modified polyethyleneimine as well as preparation method and application thereof
  • Arginine modified polyethyleneimine as well as preparation method and application thereof
  • Arginine modified polyethyleneimine as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Embodiment 1, the preparation of the polyethylenimine of arginine modification of the present invention

[0040] Arginine and polyethyleneimine with a molecular weight of 1800 are prepared according to the molar ratio of arginine and primary amino groups in polyethyleneimine being 1:2. Firstly, 0.1 mol of arginine was dissolved in 9 mL of MES buffer, 0.6 mg of EDCI / NHS was added as a catalyst, and catalyzed at 25°C for 4 h, wherein the mass ratio of EDCI and NHS was 1:2; Ethyleneimine (1mol of polyethyleneimine with a molecular weight of 1800 contains 16mol of primary amino groups) and catalyzed arginine were dissolved in 12mL N-N dimethylformamide to obtain a mixed solution, and 0.5 mL of triethylamine was used as a catalyst, and then the above reaction system was placed at 25°C and stirred for 72h to carry out the substitution reaction. After the reaction, the reaction system was transferred into a dialysis bag with a molecular weight cut-off of 1000, and dialysis wa...

Embodiment 2

[0041] Embodiment 2, the preparation of the polyethylenimine of arginine modification of the present invention

[0042] Arginine and polyethyleneimine with a molecular weight of 1800 are prepared according to the molar ratio of arginine and primary amino groups in polyethyleneimine being 1:2.5. Firstly, 0.1 mol of arginine was dissolved in 9 mL of MES buffer, 0.6 mg of EDCI / NHS was added as a catalyst, and catalyzed at 25°C for 4 h, wherein the mass ratio of EDCI and NHS was 1:2; then 15.625 mmol of poly Ethyleneimine (1mol of polyethyleneimine with a molecular weight of 1800 contains 16mol of primary amino groups) and catalyzed arginine were dissolved in 12mL N-N dimethylformamide to obtain a mixed solution, and 0.5 mL of triethylamine was used as a catalyst, and then the above reaction system was placed at 25°C and stirred for 72h to carry out the substitution reaction. After the reaction, the reaction system was transferred into a dialysis bag with a molecular weight cut-o...

Embodiment 3

[0043] Embodiment 3, the preparation of the polyethylenimine of arginine modification of the present invention

[0044] Arginine and polyethyleneimine with a molecular weight of 1800 are prepared according to the molar ratio of arginine and primary amino groups in polyethyleneimine being 1:3. First, 0.1 mol of arginine was dissolved in 10 mL of MES buffer, 1.0 mg of EDCI / NHS was added as a catalyst, and catalyzed at 25°C for 4 h, wherein the mass ratio of EDCI and NHS was 1:1.5; then 18.75 mmol of poly Ethyleneimine (1mol of polyethyleneimine with a molecular weight of 1800 contains 16mol of primary amino groups) and catalyzed arginine were dissolved in 12mL N-N dimethylformamide to obtain a mixed solution, and 0.5 mL of triethylamine was used as a catalyst, and then the above reaction system was placed at 25°C and stirred for 72h to carry out the substitution reaction. After the reaction, the reaction system was transferred into a dialysis bag with a molecular weight cut-off o...

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Abstract

The invention provides arginine modified polyethyleneimine. The arginine modified polyethyleneimine is obtained by replacing primary amine hydrogen of primary amine groups in polyethyleneimine with arginine. The molecular weight of the polyethyleneimine is less than 10000. As a gene vector, the arginine modified polyethyleneimine prepared by the invention has good plasmid DNA loading capacity, very low cytotoxicity and very high gene transfection efficiency, the transfection efficiency is higher than that of the existing product in the presence or absence of serum, and the transfection efficiency of low-molecular-weight polyethyleneimine is remarkably improved. The arginine modified polyethyleneimine provided by the invention can be used as an excellent gene vector.

Description

technical field [0001] The invention belongs to the field of chemistry and medicine, and in particular relates to arginine-modified polyethyleneimine and its preparation method and application. Background technique [0002] Gene therapy refers to a new type of therapy that delivers therapeutic genes to target organs or tissues through carriers to repair damaged or missing genes and exert therapeutic effects. At present, the main strategies of gene therapy include the introduction of exogenous genes such as tumor suppressor genes, suicide genes, siRNA, drug resistance genes, immune genes, and CRISPR-Cas9 system. However, exogenous genes cannot be actively incorporated into target cells, and need to be carried into cells by gene carriers. Therefore, vectors with low toxicity and high efficiency are the key to gene therapy. [0003] Gene vectors widely used at present are mainly divided into viral vectors and non-viral vectors. Viral vectors include retroviruses, adenoviruse...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G73/04C12N15/87C12N15/85
CPCC08G73/02C12N15/87C12N15/85C12N2800/107
Inventor 巩长旸吴秦洁王宁
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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