Polyethyleneimine derivative and application thereof as gene delivery carrier

A technology of polyethylenimine and its derivatives, which is applied in the direction of introducing foreign genetic material using carriers and recombinant DNA technology. The method is simple and easy to perform, and the effect of high transfection efficiency

Active Publication Date: 2011-11-23
ANHUI FENGYUAN PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Studies have shown that compared with unmodified PEI, the derivatives whose amine groups are partially substituted generally exhibit relatively high transfection ability and low toxicity, but when the substitution ratio exceeds 25%, the transfectio

Method used

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  • Polyethyleneimine derivative and application thereof as gene delivery carrier
  • Polyethyleneimine derivative and application thereof as gene delivery carrier
  • Polyethyleneimine derivative and application thereof as gene delivery carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Synthesis of Embodiment 1 Cyclic Phosphate Ester Monomer

[0054] The cyclic phosphate monomer is prepared by reacting 2-chloro-2-oxo-1,3,2-dioxaphospholane with absolute ethanol. The specific steps are as follows: After mixing 4.6g of absolute ethanol with 150mL of THF, add 10.1g of triethylamine, cool at 0°C for 20min, then add 2-chloro-2-oxo-1,3,2-dioxo Phospholane (14.8 g) in THF was reacted overnight at this temperature. After filtering, the triethylamine hydrochloride precipitate was removed, and the filtrate was concentrated and then distilled under reduced pressure to obtain the final product.

Embodiment 2

[0055] Synthesis and characterization of embodiment 2 polyethyleneimine derivatives

[0056] The preparation of polyethyleneimine derivatives was carried out under anhydrous environment. Concrete steps are as follows: 2g polyethyleneimine (M w =25000g / mol) was dissolved in 4mL dry DMSO, stirred at 50°C to fully dissolve, then 0.48g cyclic phosphate monomer was added, and the reaction stopped after 2h at this temperature, and the viscosity of the system increased significantly. After adding a small amount of methanol for dilution, the system was precipitated into excess ether, and after stirring for 3 hours, the supernatant was discarded to remove the unreacted cyclic phosphate monomer, and the precipitate was collected and dried under reduced pressure. This polymer was redissolved in deionized water and then dialyzed for two days (MWCO = 2000 Da). After the solution was lyophilized, it was stored at -80°C, polyethyleneimine derivative P1.

[0057] It was determined that the...

Embodiment 3

[0059] Embodiment 3 polyethyleneimine derivatives

[0060] The preparation of polyethyleneimine derivatives was carried out under anhydrous environment. Concrete steps are as follows: 2.0g polyethyleneimine (M w =1800g / mol) was dissolved in 4mL of dry DMSO, stirred at 40°C to fully dissolve, then 1.0g of cyclic phosphate monomer was added, and the reaction stopped after 3h at this temperature, and the viscosity of the system increased significantly. After adding a small amount of methanol for dilution, the system was precipitated into excess ether, and after stirring for 2 hours, the supernatant was discarded to remove the unreacted cyclic phosphate monomer, and the precipitate was collected and dried under reduced pressure. This polymer was redissolved in deionized water and dialyzed for two days (MWCO = 1000 Da). After the solution was lyophilized, it was stored at -80°C, polyethyleneimine derivative P2.

[0061] It has been determined that the grafting ratio of polyethyl...

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PUM

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Abstract

The invention provides a polyethyleneimine derivative and a preparation method thereof. The polyethyleneimine derivative is a cationic polymer and can be used as a gene delivery carrier. The polyethyleneimine derivative provided by the invention can be effectively combined with DNA (Deoxyribonucleic Acid), and nanoparticles with particle diameter potential can be obtained. With the polymer, the plasmid DNA of the green fluorescent protein and firefly luciferase protein can be effectively transfected into HEK293 cells; extremely high transfection efficiency is shown; and even the transfection efficiency is higher than that of the commercial polyethyleneimine. The derivative is easy and practical in synthesis method and can be subjected to mass production.

Description

technical field [0001] The present invention relates to a cationic polymer, in particular to a polyethyleneimine derivative and a carrier for gene transmission. Background technique [0002] Gene therapy refers to the introduction of the target genetic material into specific tissue cells (target cells) for proper expression through an appropriate carrier, replacing or correcting the disorder of its own gene structure or function, killing diseased cells or enhancing the body's ability to eliminate diseased cells etc., so as to achieve the purpose of treatment. The completion of modern gene technology and the human genome engineering map provides broad prospects for the use of gene molecular biology methods to treat various diseases. The study of gene therapy has penetrated into various fields of medical research and has become one of the most active fields of biotechnology today. [0003] Since nucleic acid substances are easily hydrolyzed by nucleases and have a large amou...

Claims

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Application Information

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IPC IPC(8): C08G73/04C12N15/63
Inventor 唐凌燕
Owner ANHUI FENGYUAN PHARM CO LTD
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