A kind of non-viral gene carrier material and its preparation method and application

A gene carrier, non-viral technology, applied in the field of non-viral gene carrier materials and its preparation, can solve the problems of low transfection efficiency of gene carrier, inability to prepare dendritic cell vaccines, etc., and achieve good controllability and applicability , Improving biocompatibility and DNA release ability, and the effect of mild reaction conditions

Inactive Publication Date: 2011-12-14
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the biggest obstacle in the process of dendritic cell transfection to prepare vaccines is the low transfection efficiency of various existing gene vectors, so that it is impossible to prepare highly efficient and specific dendritic cell vaccines, and there are technical problems

Method used

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  • A kind of non-viral gene carrier material and its preparation method and application
  • A kind of non-viral gene carrier material and its preparation method and application
  • A kind of non-viral gene carrier material and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034](1) 4mmol of 1,4-butanediol diacrylate and 3.3mmol of 5-amino-1-pentanol were mixed and dissolved in 4ml of chloroform, stirred and reacted at 90°C for 24 hours to obtain formula I (n=7 ~8) Unmodified polymers of structure;

[0035] 1000mg unmodified polymer and 52mg 1,3-propanediamine were mixed and dissolved in 4ml chloroform, stirred and reacted at room temperature for 12 hours to obtain a solution containing about 1050mg formula II (n=7~8) non-viral gene carrier material ;

[0036]

[0037] (2) Replace the solvent in the solution containing the non-viral gene carrier material in step (1) with dimethyl sulfoxide to prepare a 100 mg / mL solution, which is diluted to a concentration of 1 mg / ml, 2 mg / mL through pH=5.2 sodium acetate buffer solution. ml, 3mg / ml, 4mg / ml, 5mg / ml solutions, mixed with equal volumes of DNA to be transfected (target gene) diluted to 0.1mg / ml solution with pH = 7.4 phosphate buffer, incubated at room temperature for 15 minutes to prepare Fo...

Embodiment 2

[0041] (1) 3.3mmol of 1,5-pentanediol diacrylate and 4mmol of 5-amino-1-pentanol in chloroform were mixed and dissolved in 4ml, stirred and reacted at 90°C for 24 hours to obtain formula III (n= 7-8) Unmodified polymers with structure;

[0042] 1000mg of unmodified polymer and 65mg of 2,2-dimethyl-1,3-propanediamine were mixed and dissolved in 4ml of chloroform solution, stirred and reacted at room temperature for 12 hours to obtain about 1060mg of formula IV (n=7~ 8) the solution of non-viral gene carrier material;

[0043]

[0044] (2) Replace the solvent in the solution containing the non-viral gene carrier material in step (1) with dimethyl sulfoxide to prepare a 100 mg / mL solution, which is diluted to a concentration of 1 mg / ml, 2 mg / mL through pH=5.2 sodium acetate buffer solution. ml, 3mg / ml, 4mg / ml, 5mg / ml solutions, mixed with equal volumes of DNA to be transfected (target gene) diluted to 0.1mg / ml solution with pH = 7.4 phosphate buffer, incubated at room tempera...

Embodiment 3

[0047] (1) 4mmol of 1,4-butanediol diacrylate and 3.3mmol of 5-amino-1-pentanol were mixed and dissolved in 4ml of chloroform, stirred and reacted at 90°C for 24 hours to obtain the formula V (n=7 ~8) Unmodified polymers of structure;

[0048] 1000mg of unmodified polymer and 52mg of 1,3-propanediamine were mixed and dissolved in 4ml of chloroform, stirred and reacted at room temperature for 12 hours to obtain a solution containing about 1050mg of formula VI (n=7~8) non-viral gene carrier material ;

[0049]

[0050] (2) Replace the solvent in the solution containing the non-viral gene carrier material in step (1) with dimethyl sulfoxide to prepare a 100mg / mL solution, and dilute to a concentration of 1mg / ml, 2mg / ml through pH=5.2 sodium acetate buffer solution , 3mg / ml, 4mg / ml, 5mg / ml solutions, mixed with equal volumes of DNA to be transfected (target gene) diluted to 0.1mg / ml solution with phosphate buffer solution of pH = 7.4, and incubated at room temperature for 15 m...

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Abstract

The invention discloses a preparation method of a non-viral gene vector material. The method comprises the following steps of: performing michael reaction on polyol acrylate and amino alcohol to prepare an unmodified polymer; and reacting the unmodified polymer with a diamino group containing compound to prepare an ester bond-containing non-viral gene vector material. The method has the characteristics of easiness in preparation and high controllability; and the prepared non-viral gene vector material has the characteristics of high efficiency and low toxicity. The invention discloses application of the non-viral gene vector material to transfection of cells; and the application comprises the following steps of: mixing the non-viral gene vector material and genes; incubating to prepare a compound of a non-viral gene vector and a gene; and adding the prepared compound of the non-viral gene vector and the gene into receptor cells to finish the transfection of the genes in the cells. Thecompound of the non-viral gene vector and the gene has high gene transfection efficiency and superior biological safety for the transfection of the cells.

Description

technical field [0001] The present invention relates to the field of gene carrier, the preparation of gene carrier and the application field of gene carrier transfection, in particular to a kind of non-viral gene carrier material and its preparation method and the preparation of the compound of non-viral carrier and gene to tumor cells, bone marrow Application of gene transfection in cells such as mesenchymal stem cells and dendritic cells. Background technique [0002] Gene therapy refers to the introduction of exogenous normal genes into target cells to correct or compensate diseases caused by gene defects and abnormalities, so as to achieve therapeutic purposes. That is, the exogenous gene is inserted into the recipient cell through gene transfer technology, so that the product produced by the exogenous gene can treat a certain disease. A key task in gene therapy is to find a safe and efficient gene transfer technology, that is, the construction of gene transfer vectors....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G73/00C07C229/16C07C227/08C12N15/87
Inventor 高建青陈煜哲姚醒蕾
Owner ZHEJIANG UNIV
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