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High-performance cationic gene vectors with PGMA (polyglycidyl methacrylate) serving as framework constructed by ATRP (atom transfer radical polymerization) method

A gene carrier and cation technology, applied in the field of cationic gene carriers, can solve the problems of different vector transfection efficiency, different protonation ability, etc., and achieve the effects of simple use method, easy regulation and good storage stability.

Inactive Publication Date: 2012-08-22
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] 2. When preparing high-performance cationic gene carriers, different monomers can be grafted to obtain cationic polymers with different properties, but the protonation ability of different monomers is different, resulting in different transfection efficiencies of the carriers. High-performance monomers are issues that need to be considered

Method used

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  • High-performance cationic gene vectors with PGMA (polyglycidyl methacrylate) serving as framework constructed by ATRP (atom transfer radical polymerization) method
  • High-performance cationic gene vectors with PGMA (polyglycidyl methacrylate) serving as framework constructed by ATRP (atom transfer radical polymerization) method
  • High-performance cationic gene vectors with PGMA (polyglycidyl methacrylate) serving as framework constructed by ATRP (atom transfer radical polymerization) method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1) Continuously react under nitrogen protection at 50℃, add 12g GMA (glycidyl methacrylate) into a small flask, then add 10g THF (tetrahydrofuran), 120mg pentamethyldiethylenetriamine, 213.6mg PMDETA, and finally Add 86.4mg CuBr to initiate active controllable free radical polymerization; after 3h, open the stopper and accelerate the stirring for 10min, fully contact with air to stop the reaction. The polymerized product is repeatedly precipitated with methanol until the morphology becomes solid, and then placed in a vacuum drying box to remove the methanol , The linear PGMA is obtained, the number average molecular weight (Mn) of the polymer is 580000 g / mol, and the PDI (Mw / Mn) is 1.23.

[0035] 2) Continuous reaction under nitrogen protection at 50℃, add 0.3g of linear PGMA obtained in step 1) to 5.6g of THF to dissolve, and then add 3g of ethanolamine (or use 2-amino-1-propanol, 3-amino-1- Propanol, NN-dimethylethylenediamine, or 0.15g ethanolamine and 0.15g NN-dimethyl...

Embodiment 2

[0037] 1) Continue the reaction under nitrogen protection at 37°C. Take 1.5 g of the linear PGMA obtained in step 1) of Example 1 in a flask and dissolve it in 10 g of THF, add 0.36 g of BIBA (2-Bromoisobutyric acid), and react for 24 hours. , Ether precipitation, vacuum drying to obtain PGMA-Br (PGMA / BIBA is 5:1, one of every 6 GMA segments on the molecular chain is connected with Br).

[0038] 2) Continuous reaction under nitrogen protection at 50℃, add 0.3g of PGMA-Br obtained in step 1) above to 5g THF to dissolve, then add 4gGMA, 82mg HMTETA, and finally 33.5mg CuBr to initiate active controllable free radicals Polymerization, after 4 hours of reaction, open the stopper and accelerate the stirring for 10 minutes, and fully contact with the air to stop the reaction; the polymerization product is repeatedly precipitated with methanol until the product becomes solid, and the comb-shaped PGMA is obtained after the methanol is removed in a vacuum drying cabinet.

[0039] 3) Continu...

Embodiment 3

[0041] 1) Continuous reaction at 37°C under nitrogen protection, take 2.5 g of the linear PGMA obtained in step 1) of Example 1 in a flask and dissolve it in 10 g of THF, add 0.37 g of BIBA (2-Bromoisobutyric acid) and react for 24 hours. Ether precipitation, vacuum drying to obtain PGMA-Br (PGMA / BIBA is 8:1, one of every 8 GMA segments on the molecular chain is connected with Br).

[0042] 2) Continuous reaction under nitrogen protection at 50℃, add 0.3g of PGMA-Br obtained in step 1) above to 5g THF to dissolve, then add 4gGMA, 82mg HMTETA, and finally 33.5mg CuBr to initiate active controllable free radicals Polymerization, after 4 hours of reaction, open the stopper and accelerate the stirring for 10 minutes, and fully contact with the air to stop the reaction; the polymerization product is repeatedly precipitated with methanol until the product becomes solid, and the comb-shaped PGMA is obtained after the methanol is removed in a vacuum drying cabinet.

[0043] 3) Continuous r...

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Abstract

The invention discloses a series of low-toxicity and efficient cationic gene vectors with PGMA (polyglycidyl methacrylate) serving as a framework constructed by an ATRP (atom transfer radical polymerization) method, which belong to the technical field of nonviral gene vectors. The ATRP method is stable in polymerization reaction and easy in regulation and control, and can prepare various high-performance cationic gene vectors different in molecular weight and narrow molecular weight distribution according to needs. The prepared cationic gene vectors are high in storage stability, higher than gold mark PEI (polyethyleneimine) in transfection efficiency in cells such as Hepg2, C6, Cos7, HEK293 and the like and simple in use method, and have commercial potential.

Description

Technical field [0001] The invention belongs to the technical field of non-viral gene carriers, and specifically relates to the ATRP method to construct a series of cationic gene carriers with PGMA (polyglycidyl methacrylate) as the backbone and high gene transfection efficiency and low toxicity. Background technique [0002] Gene therapy methods provide a promising new approach for the treatment of congenital genetic diseases and serious acquired diseases. Gene therapy is a treatment method that introduces foreign genes into target cells and effectively expresses them to achieve the purpose of curing diseases. DNA molecules usually exist in a loose, negatively charged state and are large in size. There is a repulsive effect between the full negative charge and the cell surface, which makes it difficult to enter the cell. In addition, there are a large number of hydrolytic enzymes, especially nucleases, in the blood and cells, which can remove the naked body. DNA molecules are d...

Claims

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Application Information

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IPC IPC(8): C08F120/32C08F8/32C08F293/00C08F220/32C08F220/34C08F220/54C08F220/28C08F299/02C12N15/63
Inventor 徐福建柴明英杨鑫超王增辉朱韵修可茂胡杨
Owner BEIJING UNIV OF CHEM TECH
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