A modified polyhydroxyethylacrylamide transgenic carrier and its preparation method and application

A technology of polyhydroxyethylacrylamide and transgenic vectors, which is applied in the direction of using vectors to introduce foreign genetic material, recombinant DNA technology, etc., and can solve the problems of low biodegradability, low transfection efficiency, toxicity, etc.

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

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Problems solved by technology

Therefore, cationic polymers have become an important part of non-viral gene carriers, but when cationic polymers are used as gene carriers, there are still some problems, such as poor biocompatibility, toxicity, low biodegradability and transfection low efficiency

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  • A modified polyhydroxyethylacrylamide transgenic carrier and its preparation method and application
  • A modified polyhydroxyethylacrylamide transgenic carrier and its preparation method and application
  • A modified polyhydroxyethylacrylamide transgenic carrier and its preparation method and application

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Experimental program
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Effect test

Embodiment 1

[0031] Monomer HEAA (5 mmol, 575.5 mg) and catalyst CuCl (9.9 mg, 0.1 mmol) were added to a Schlenk tube and dissolved with 5 mL of ethanol / water. Freezing with liquid nitrogen, degassing, then dissolving and blowing nitrogen gas, repeated freezing and thawing three times to remove oxygen in the reaction system. Initiator PMDETA (21 ml, 0.1 mmol) and ligand EBMP (15 ml, 0.1 mmol) were dissolved in 1 ml ethanol / water solution and transferred to a Schlenk tube with a syringe under nitrogen protection. After the reaction mixture was repeatedly frozen and thawed three times, it was vacuum-sealed, stirred and reacted in a water bath at 60°C for 10 hours, the reaction product was diluted with deionized water, and then dialyzed in deionized water with a dialysis bag (molecular weight cut-off 3500), and the water was changed every 8 hours , lyophilized after 5 days of dialysis. Polyhydroxyethylacrylamide (PHEAA, theoretical degree of polymerization is 50) can be obtained;

[0032] W...

Embodiment 2

[0037] Monomer HEAA (10 mmol, 1.151 g) and catalyst CuCl (9.9 mg, 0.1 mmol) were added to a Schlenk tube and dissolved with 8 mL of ethanol / water. Freezing with liquid nitrogen, degassing, then dissolving and blowing nitrogen gas, repeated freezing and thawing three times to remove oxygen in the reaction system. Ligand PMDETA (21 ml, 0.1 mmol) and initiator EBMP (15 ml, 0.1 mmol) were dissolved in 1 ml ethanol / water solution, and transferred to a Schlenk tube with a syringe under nitrogen protection. After the reaction mixture was repeatedly frozen and thawed three times, it was vacuum-sealed, stirred and reacted in a water bath at 60°C for 10 hours, the reaction product was diluted with deionized water, and then dialyzed in deionized water with a dialysis bag (molecular weight cut-off 3500), and the water was changed every 8 hours , lyophilized after 5 days of dialysis. Polyhydroxyethylacrylamide (PHEAA, theoretical degree of polymerization is 100) can be obtained;

[0038]...

Embodiment 3

[0043] Monomer HEAA (15 mmol, 1.7265 g) and catalyst CuCl (9.9 mg, 0.1 mmol) were added to a Schlenk tube and dissolved with 11 mL of ethanol / water. Freezing with liquid nitrogen, degassing, then dissolving and blowing nitrogen gas, repeated freezing and thawing three times to remove oxygen in the reaction system. Ligand PMDETA (21 ml, 0.1 mmol) and initiator EBMP (15 ml, 0.1 mmol) were dissolved in 1 ml ethanol / water solution, and transferred to a Schlenk tube with a syringe under nitrogen protection. After the reaction mixture was repeatedly frozen and thawed three times, it was vacuum-sealed, stirred and reacted in a water bath at 60°C for 10 hours, the reaction product was diluted with deionized water, and then dialyzed in deionized water with a dialysis bag (molecular weight cut-off 3500), and the water was changed every 8 hours , lyophilized after 5 days of dialysis. Polyhydroxyethylacrylamide (PHEAA, theoretical degree of polymerization is 150) can be obtained;

[004...

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Abstract

The invention discloses a modified polyhydroxyethyl acrylamide transgenic vector. Polyhydroxyethyl acrylamide is taken as a framework structure; and a side-chain substituent R of the polyhydroxyethyl acrylamide is NH2(CH2)n1N(CH2)n2NH2, wherein n1 is 1 to 5 and n2 is 1 to 5. A preparation method comprises the following steps of: preparing the polyhydroxyethyl acrylamide through atom transfer radical polymerization, protecting primary amine of a small molecular modifier by using trifluoroacetic acid ethyl ester, grafting the primary amine onto the polyhydroxyethyl acrylamide, and finally performing deprotection to obtain the transgenic vector. Cationized polyhydroxyethyl acrylamide is convenient for cell internalization and can protect deoxyribonucleic acid (DNA) from being degraded by DNase, so that the vector can enter cells, and then the DNA is released.

Description

technical field [0001] The invention relates to a transgene carrier and a preparation method thereof, more specifically, to a modified polyhydroxyethylacrylamide transgene carrier and a preparation method thereof. Background technique [0002] Gene therapy refers to the introduction of human normal genes or therapeutic genes into human target cells in a certain way to correct gene defects or exert therapeutic effects. The vectors used in gene therapy mainly include viral vectors (viral vector) and nonviral vectors (nonviral vector). Viral vectors have made some breakthroughs in gene therapy, but in the process of clinical application, their immunogenicity and potential carcinogenicity are still major hidden dangers that are difficult to overcome. As another way of gene delivery, non-viral vectors have been widely studied, including naked DNA, liposomes, and cationic polymers. [0003] As a kind of non-viral vector, cationic polymer not only has the characteristics of non-v...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C08F120/58C08F8/38C08F8/32
Inventor 刘文广王宏博杨建海王玮
Owner TIANJIN UNIV
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