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Non-virogene transfection carrier, complex particles of the same and plasmid DNA, preparing method and using method

A gene transfection carrier and non-viral technology, which is applied in the field of non-viral gene transfection carrier, its and plasmid DNA complex particles and preparation, can solve the problems of low transfection efficiency and high cytotoxicity of PEI, and improve the efficiency of transfection. Infection efficiency, reduced cytotoxicity, and low cytotoxicity

Inactive Publication Date: 2007-12-12
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are still two outstanding problems with PEI as a gene carrier
(1) Compared with viral vectors, its transfection efficiency is not high; (2) PEI cytotoxicity is relatively high

Method used

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  • Non-virogene transfection carrier, complex particles of the same and plasmid DNA, preparing method and using method
  • Non-virogene transfection carrier, complex particles of the same and plasmid DNA, preparing method and using method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Preparation of polyethyleneimine-γ-benzyl ester-L-glutamic acid copolymer

[0035] Take 1g of hyperbranched polyethyleneimine with molecular weights of 1.8K, 5K, 10K, 25K, and add 100 mL of chloroform to dissolve it.

[0036] The N carboxylic anhydride cyclic monomer of γ-benster-L-glutamic acid is dissolved in chloroform to prepare a chloroform solution with a concentration of 1% by weight. Under magnetic stirring, according to the mass ratio of the N carboxylic anhydride cyclic monomer of polyethyleneimine and γ-benster-L-glutamic acid of 1:9 to 4:6, the γ-benster-L- The chloroform solution of the N carboxylic anhydride cyclic monomer of glutamic acid was added dropwise to the chloroform solution of polyethyleneimine, and the addition was completed within 3 hours, and the reaction was carried out at room temperature for 72 hours. After the reaction is complete, the reactant is settled out with ether and dried in vacuum. The product was then dissolved in seconda...

Embodiment 2

[0039] Example 2: Preparation of polyethyleneimine-ε-benzyloxycarbonyl lysine copolymer

[0040] Take 1 g of hyperbranched polyethyleneimine with molecular weights of 1.8K, 5K, 10K, 25K, and add 100 mL of chloroform to dissolve it.

[0041] The N carboxylic anhydride cyclic monomer of ε-benzyloxycarbonyl lysine was dissolved in chloroform to prepare a chloroform solution with a concentration of 1% by weight. Under magnetic stirring, according to the mass ratio of the N carboxylic anhydride cyclic monomer of polyethyleneimine and ε-benzyloxycarbonyl lysine of 1:9 to 4:6, the N carboxylic acid of ε-benzyloxycarbonyl lysine The chloroform solution of the acid anhydride cyclic monomer was added dropwise to the chloroform solution of polyethyleneimine, and the addition was completed within 3 hours, and the reaction was carried out at room temperature for 72 hours. After the reaction was completed, the reactant was settled out with ether and dried in vacuum. The product was then dissolv...

Embodiment 3

[0044] Example 3: Preparation of polyethyleneimine-phenylalanine copolymer

[0045] Take 1 g of hyperbranched polyethyleneimine with molecular weights of 1.8K, 5K, 10K, 25K, and add 100 mL of chloroform to dissolve it. The N carboxylic anhydride cyclic monomer of phenylalanine was dissolved in chloroform to prepare a chloroform solution with a concentration of 1% by weight. Under magnetic stirring, according to the mass ratio of the N carboxylic anhydride cyclic monomer of polyethyleneimine and γ-benster-L-glutamic acid of 1:9 to 4:6, the γ-benster-L- The chloroform solution of the N carboxylic anhydride cyclic monomer of glutamic acid was added dropwise to the chloroform solution of polyethyleneimine, and the addition was completed within 3 hours, and the reaction was carried out at room temperature for 72 hours. After the reaction was completed, the reactant was settled out with ether and dried in vacuum. The product was then dissolved in secondary water, and the secondary aqueo...

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Abstract

The invention belongs to technology field of carrier material for non-viral gene transfection, and relates to a carrier material for non-viral gene transfection, plasmid DNA compound particle and preparation method and administration. The weight ratio of polyethylene imine and hydrophobic unit containing benzene ring is 1:9-4:6.The polyethylene imine is superbranched polyethylene imine (PEI) with average molecular weight of 1.8K-25K.The hydrophobic unit containing benzene ring is one or more of gamma -benzyl-L-glutamic acid, epsi -carbobenzoxy lysine and phenylalanine at any ratio. The water soluble polyethylene imine modified substance of hydrophobic unit containing benzene ring can compound with genic material to form a particle with positive charge on surface and particle diameter of 50-300nm.The compound particle can transfer loaded genic material into cell to realize expression of genic material and complete transfection process. The invention prepared non-gene transfection carrier material has the advantages of low toxicity and high up transfection efficiency; and can be used as transfection agent in vivo and in vitro.

Description

Technical field [0001] The invention belongs to the technical field of non-viral gene transfection vectors, and relates to a non-viral gene transfection vector, its and plasmid DNA complex particles, and a preparation method and usage. Background technique [0002] Gene therapy refers to a treatment method that uses DNA transfer to treat or even prevent human diseases. In recent years, the development of gene therapy has made it possible to treat genetic diseases or cancer. Successful gene therapy relies on effective gene vectors. Using viral vectors to mediate gene transfer is the most widely used method in gene therapy, including retrovirus, adenovirus (AV), adeno-associated virus (AAV), herpes simplex virus (HSV), vaccinia virus (VV) and other vectors . However, viral vectors have great safety risks in clinical applications. Non-viral vectors have become the most promising alternatives to viral vectors due to their advantages such as safety, effectiveness, and non-immunogenici...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/34A61K48/00A61K9/16C12N15/63C08F126/02C08F8/46C08F8/30A61P35/00A61P43/00
Inventor 陈学思田华雨陈磊王宇夏加亮陈杰景遐斌
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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