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ATRP (Atom Transfer Radical Polymerization) method for constructing high-transfection niosomes cation gene carrier

A gene carrier and cationic technology, which is applied in the field of non-viral gene carrier preparation, can solve the problems of low effective toxicity, high toxicity, and inequalities, and achieve good transfection efficiency and increase transfection efficiency

Active Publication Date: 2015-10-07
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] There are still many problems in the research process of using activity-controlled polymerized ATRP to obtain a series of liposome-like cationic gene carriers. For example, this kind of cationic carrier has higher transfection efficiency with the increase of molecular weight, but the toxicity How to increase the transfection efficiency while ensuring moderate toxicity has become the focus of attention; different monomers have different characteristics, some have higher transfection efficiency and lower effective toxicity, how to screen out high-efficiency and high-performance Monomer is also a problem that people need to consider; of course, the same liposome gene carrier has different transfection efficiency for different cell lines, how to choose the most suitable gene carrier for different cells is also a problem that needs to be studied

Method used

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  • ATRP (Atom Transfer Radical Polymerization) method for constructing high-transfection niosomes cation gene carrier
  • ATRP (Atom Transfer Radical Polymerization) method for constructing high-transfection niosomes cation gene carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Take 0.2g of cholesterol initiator CHO-Br completely dissolved in 5mL of dimethyl sulfoxide (DMSO), then 1.73mL of glycidyl methacrylate (GMA) and 135μL of pentamethyldiethylenetriamine (PMDETA) Dissolve it in the solution, and quickly add 54 mg of cuprous bromide (CuBr) after bubbling nitrogen gas for 5 minutes, and immediately seal it with a rubber stopper. The system was reacted at 27-30°C under an oxygen-free nitrogen protection environment for 30 minutes, opened the cork and stood still for 2-3 minutes, then precipitated with methanol, and washed twice with methanol to remove PDMETA, CuBr, unreacted GMA and large Part of the solvent DMSO. After centrifugation, the supernatant was removed, and the residual methanol in the lower precipitate was dried, then dissolved with 1-2mL DMSO, precipitated with deionized water and washed twice. After centrifugation, the supernatant was removed, and a small amount of pure water was added to lyophilize. The freeze-dried product...

Embodiment 2

[0037] Take 0.2 g of cholesterol initiator, the reaction steps and post-treatment are as in Example 1, except that the volume of glycidyl methacrylate (GMA) added is 3.46 mL. The polymer obtained in this way is designated as CHO-PGMA2. The number average molecular weight (Mn) of the polymer (CHO-PGMA2) was 9365, and the molecular weight distribution index (Mw / Mn) was 1.23.

[0038] Take 0.15 g of the obtained CHO-PGMA2 and dissolve it in 4 mL of DMSO. The reaction steps and post-treatment methods are as in Example 1. The obtained flocculent polymer is the cationic gene carrier of the cholesterol matrix we need, which is recorded as CHO-PGEA2.

[0039] The number average molecular weight (Mn) of the polymer (CHO-PGEA2) was 15000, and the molecular weight distribution index (Mw / Mn) was 1.85.

Embodiment 3

[0041] Take 0.2 g of cholesterol initiator, the reaction steps and post-treatment are as in Example 1, except that the volume of glycidyl methacrylate (GMA) added is 5.19 mL. The polymer obtained in this way is designated as CHO-PGMA3. The number average molecular weight (Mn) of the polymer (CHO-PGMA3) was 13500, and the molecular weight distribution index (Mw / Mn) was 1.30.

[0042] Take 0.15 g of the obtained CHO-PGMA3 and dissolve it in 4 mL of DMSO. The reaction steps and post-treatment methods are as in Example 1. The obtained flocculent polymer is the cationic gene carrier of the cholesterol matrix we need, which is recorded as CHO-PGEA3.

[0043] The number average molecular weight (Mn) of the polymer (CHO-PGEA3) was 18000, and the molecular weight distribution index (Mw / Mn) was 1.80.

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Abstract

The invention discloses a cation gene carrier-constructed niosomes cation gene carrier with excellent performance such as high transfection and low toxicity, obtained by combining a commonly used activity polymerization method based on lipids (cholesterol, phosphatidylinositol and the like). A method is easy and feasible, a polymerization method ensures effective controllable molecular weight, compared with a common cation gene carrier, the gene carrier has a characteristic of being effectively compatible with a cytomembrane by a lipid matrix, or a characteristic of establishing into lipidosome per se to increase intake of cells, so the transfection efficiency of the cells is increased. The cation gene carrier of a series of niosomes constructed by the method has well transfection efficiency in cell lines of HepG2, COS7, C6, SL, H9C2 and the like, the transfection efficiency is higher than the transfection efficiency of the internationally commercially used lipidosome lipfect2000, and the niosomes cation gene carrier effectively solves the difficult problem of poorer in-vivo transfection efficiency of the lipidosome lipfect2000, and has higher commercial potential.

Description

technical field [0001] The invention belongs to the technical field of non-viral gene carrier preparation, and specifically relates to an ATRP method to construct a series of liposome-like cationic gene carriers with high gene transfection efficiency using lipids (cholesterol, phosphatidylinositol, etc.) as substrates. Background technique [0002] Gene therapy has been recognized as a new and effective treatment method in the past two decades. This method plays a pivotal role in future medical treatment and is widely used to treat a series of genetic diseases, human cancer and cardiovascular diseases, etc. Wait. In a broad sense, the whole gene therapy is to introduce the exogenous design gene or oligonucleotide fragment used for treatment into the diseased cells in the disease body through the gene carrier, so as to express and improve the defects of the diseased cells, so as to achieve the purpose of treatment. The entire process of gene therapy has three important steps...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F120/32C08F120/34C08F8/32C12N15/87
Inventor 徐福建许晨俞丙然
Owner BEIJING UNIV OF CHEM TECH
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