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ATRP method for constructing cationic gene vector with PGMA as skeleton

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

Inactive Publication Date: 2012-10-03
萨恩化学技术(上海)有限公司
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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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  • ATRP method for constructing cationic gene vector with PGMA as skeleton
  • ATRP method for constructing cationic gene vector with PGMA as skeleton
  • ATRP method for constructing cationic gene vector with PGMA as skeleton

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1) Continuous reaction under nitrogen protection conditions at 50°C, add 12g of GMA (glycidyl methacrylate) into a small flask, then add 10g of THF (tetrahydrofuran), 120mg of pentamethyldiethylenetriamine, 213.6mg of PMDETA, and finally Add 86.4mg of CuBr to initiate active controllable free radical polymerization; after 3h, open the bottle stopper to accelerate stirring for 10min, fully contact with air to stop the reaction, and the polymerization product is repeatedly precipitated with methanol until the shape becomes solid, then put it in a vacuum drying oven to remove methanol , that is, linear PGMA is obtained, the number average molecular weight (Mn) of the polymer is 580000g / mol, and the PDI (Mw / Mn) is 1.23.

[0036] 2) Continuous reaction under nitrogen protection at 50°C, add 0.3g of linear PGMA obtained in step 1) to 5.6g of THF to dissolve, then add 3g of ethanolamine (or use 2-amino-1-propanol, 3-amino-1- Propanol, N-N-dimethylethylenediamine, or 0.15g etha...

Embodiment 2

[0038] 1) Continuous 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, and vacuum drying to obtain PGMA-Br (PGMA / BIBA is 5:1, and one of every six GMA segments on the molecular chain is connected to Br).

[0039] 2) Continuous reaction under nitrogen protection at 50°C, add 0.3g of PGMA-Br obtained in the above step 1) into 5g of THF to dissolve, then add 4gGMA, 82mg of HMTETA in sequence, and finally add 33.5mg of CuBr to trigger active controllable free radicals Polymerization, after 4 hours of reaction, open the bottle stopper to accelerate stirring for 10 minutes, and fully contact with air to stop the reaction; the polymerization product is repeatedly precipitated with methanol until the product becomes solid, and then placed in a vacuum drying oven to remove methanol to obtain comb-sh...

Embodiment 3

[0042] 1) Continuous reaction under nitrogen protection at 37°C. 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) to react for 24 hours. Precipitate with ether and dry in vacuum to get PGMA-Br (PGMA / BIBA ratio is 8:1, one of every 8 GMA segments on the molecular chain is connected with Br).

[0043] 2) Continuous reaction under nitrogen protection at 50°C, add 0.3g of PGMA-Br obtained in the above step 1) into 5g of THF to dissolve, then add 4gGMA, 82mg of HMTETA in sequence, and finally add 33.5mg of CuBr to trigger active controllable free radicals Polymerization, after 4 hours of reaction, open the bottle stopper to accelerate stirring for 10 minutes, and fully contact with air to stop the reaction; the polymerization product is repeatedly precipitated with methanol until the product becomes solid, and then placed in a vacuum drying oven to remove methanol to obtain comb-shap...

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Abstract

The invention discloses an ATRP method for constructing a series of low-toxicity and high-efficiency cationic gene vectors with PGMA (polyglyceryl methacrylate) as a skeleton in the technical field of the non-viral gene vectors. The ATRP method is smooth in polymerization reaction and easy to control; according to the ATRP method, a variety of high-performance cationic gene vectors with different molecular weights and narrow molecular weight distribution can be prepared as required; and the prepared cationic gene vectors are good in storage stability, have a transfection efficiency higher than the gold standard PEI in Hepg2, C6, Cos7, HEK293 and other cells, are easy to use and have a 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 skeleton, high gene transfection efficiency and low toxicity. [Background technique] [0002] Gene therapy approaches offer a promising new avenue for the treatment of congenital genetic diseases and severe acquired diseases. Gene therapy is a treatment method that introduces foreign genes into target cells and expresses them effectively, so as to achieve the purpose of curing diseases. DNA molecules usually exist in a negatively charged loose state, with a large volume, and there is a repulsion between the full negative charge and the cell surface, making 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 The DNA molecule is damaged, so transfect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F120/32C08F8/32C08F265/04C12N15/87
Inventor 李展孙文海罗波
Owner 萨恩化学技术(上海)有限公司
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