Lipopolysaccharide amine cationic polymer and preparation method and application thereof

A cationic polymer and lipopolysaccharide technology, applied in the field of biomedicine, can solve the problems of low gene transfection efficiency, virus transfection level, low transfection efficiency, etc., and achieve excellent biocompatibility, reduce cytotoxicity, and low toxicity Effect

Inactive Publication Date: 2014-06-18
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the gene transfection efficiency of this vector is still much lower than that of virus
When using this carrier to carry out in vitro serum-free transfection to mouse Jurkat cells, the highest gene transfection efficiency of its green fluorescent protein (GFP) can only reach about 18%; and when there is serum transfection, its transfection efficiency is even higher less than 1%

Method used

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  • Lipopolysaccharide amine cationic polymer and preparation method and application thereof
  • Lipopolysaccharide amine cationic polymer and preparation method and application thereof
  • Lipopolysaccharide amine cationic polymer and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1 Synthesis of reduced lipopolysaccharide amine cationic polymer rMASA-PEI-Cho

[0044] 1.1 Synthesis of cholesterol-terminated polyethyleneimine (PEI-Cho)

[0045] First, the methylene chloride and PEI with a molecular weight of less than 2k are dehydrated, and then 3g PEI (1.67×10 -3 mol), 10mL dichloromethane, 100μL triethylamine (7.17×10 -3 mol) fully stirred and mixed on ice for 30min to obtain a clear and transparent solution A; take 0.75g cholesterol chloroformate (1.67×10 -3 mol, where the molar feed ratio of PEI to cholesterol chloroformate is 1:1. ) Dissolve in 5 mL of ice-cold anhydrous dichloromethane to obtain a clear and clear solution B. Under stirring, solution 2 was slowly added dropwise to solution A within 30 minutes, and then the reaction mixture was allowed to continue stirring and reacting for 12 hours on ice, and the solvent was removed by rotary evaporation to obtain a white extremely viscous semi-solid. The semi-solid was dissolved in 50 mL of 0.1 m...

Embodiment 2

[0054] 1 Synthesis of lipopolysaccharide amine cationic graft copolymer MASA-PEI-Cho

[0055] 1.1 Synthesis of cholesterol-terminated polyethyleneimine (PEI-Cho)

[0056] First, the methylene chloride and PEI are dehydrated, and then 3g PEI (1.67×10 -3 mol), 10mL dichloromethane, 200μL triethylamine (1.43×10 -2 mol) Stir and mix well on ice for 30 minutes to obtain a clear and transparent solution A; take 1.5 g of cholesterol chloroformate (its molar ratio to PEI is 2:1) and dissolve it in 10 mL of ice-cold anhydrous dichloromethane to obtain a clear and clear solution B. Under stirring, solution B was slowly added dropwise to solution A within 30 minutes, and then the reaction mixture was allowed to continue stirring and reacting for 12 hours on ice, and the solvent was removed by rotary evaporation to obtain a white extremely viscous semi-solid. The semi-solid was dissolved in 50 mL 0.5 mol / L HCl, filtered, and the filtrate was extracted 3 times with 100 mL dichloromethane to re...

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Abstract

The invention discloses a lipopolysaccharide amine cationic polymer, which is synthesized with hydrophilic polyaldehyde-based sodium alginate is adopted as a main chain, hydrophobic cholesterol capped polyethylene imine of low molecular weight as a side chain. The physical and chemical structure, buffering capacity, transfection efficiency and cytotoxicity of the lipopolysaccharide amine cationic polymer are characterized. According to exploration of the feasibility of use as a gene carrier and the effects on improvement of the gene transfection efficiency and reduction of the cytotoxicity, the polymer has advantages of good biocompatibility, controllable degradation performance, low cytotoxicity, high safety and practicality and low cost, and the transfection efficiency of GFP (green fluorescent protein) in MSCs (mesenchymal stem cells) is higher than 95 percent regardless of the presence or absence of serum. Moreover, the polymer can be used as a gene carrier and other drug carriers such as anticancer drugs, RNA (ribose nucleic acid) carriers and the like. The invention also discloses a preparation method and application of the polymer.

Description

Technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a lipopolysaccharide amine cationic polymer and a preparation method and application thereof. Background technique [0002] Gene therapy will provide unlimited possibilities for the treatment of all gene-related diseases, so it has become a research hotspot in the past two decades. The bottleneck of gene therapy in clinical application is its safety and effectiveness. Since viral vectors have many unpredictable and unsolvable safety hazards such as immunogenicity, toxicity, carcinogenicity, host DNA insertion and integration, non-viral vectors have become a hope for the clinical application of gene therapy. In the past ten years, people have begun to devote themselves to designing various new non-viral vectors, studying ways to improve their transfection efficiency while reducing cytotoxicity, and have made great progress, but there is still a certain gap between the t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G81/00C08G73/04C08B37/04A61K48/00A61K47/36C12N15/87
Inventor 王琴梅滕伟
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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