Star-shaped multi-targeting functional gene vector based on POSS and application

A gene carrier and functional technology, applied in the field of star-shaped multi-targeting functional gene carriers, can solve the problems of lack of nuclear localization ability, affecting the delivery effect, etc., so as to facilitate the uptake of cells, improve the effect of gene delivery, and enhance the load gene the effect of the ability

Active Publication Date: 2018-09-14
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported in the literature that linking cell-targeting functional peptides to polymers improves the specificity of gene delivery systems, but they lack nuclear localization, which affects its delivery efficacy

Method used

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  • Star-shaped multi-targeting functional gene vector based on POSS and application
  • Star-shaped multi-targeting functional gene vector based on POSS and application
  • Star-shaped multi-targeting functional gene vector based on POSS and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The preparation method of the star-shaped multi-targeting functional gene carrier based on POSS comprises the steps:

[0048] (1) octaamino POSS is dissolved in high-purity water, and NaOH aqueous solution is added for the first time to obtain solution one, and the mol ratio of NaOH in the described octaamino POSS and the NaOH aqueous solution added for the first time is 1:4; Dissolve isobutyryl bromide in dichloromethane to obtain 2-bromoisobutyryl bromide solution; while adding 2-bromoisobutyryl bromide solution to solution 1, add NaOH aqueous solution for the second time, and stir at 0°C Down reaction 10 hours, obtain macromolecular initiator POSS-(Br) 4 The crude product is refined, and the refining steps are: successively extract with HCl aqueous solution and NaOH aqueous solution, separate the organic layer, and then dry it with anhydrous sodium sulfate for 18 hours; obtain a concentrated solution by rotary evaporation after filtration, and dissolve the concentrat...

Embodiment 2

[0064] The preparation method of the star-shaped multi-targeting functional gene carrier based on POSS comprises the steps:

[0065] (1) octaamino POSS is dissolved in high-purity water, and NaOH aqueous solution is added for the first time to obtain solution one, and the mol ratio of NaOH in the described octaamino POSS and the NaOH aqueous solution added for the first time is 1:9; Dissolve isobutyryl bromide in chloroform to obtain 2-bromoisobutyryl bromide solution; while adding 2-bromoisobutyryl bromide solution to solution 1, add NaOH aqueous solution for the second time, and stir at 5°C Down reaction 18 hours, obtain macromolecular initiator POSS-(Br) 8 The crude product, refining, the method for refining is with embodiment 1, obtains macromolecular initiator POSS-(Br) 8 ; The mol ratio of the octaamino POSS, 2-bromoisobutyryl bromide and the NaOH that adds the NaOH aqueous solution for the second time is 1:24:48;

[0066] The octaamino POSS is the abbreviation of octa...

Embodiment 3

[0081] The control is polyethyleneimine / ZNF580 gene complex (PEI 25kDa / DNA complex), wherein the mass ratio of PEI 25kDa to ZNF580 gene is 1:2;

[0082] Polyethyleneimine (PEI 25kDa) was purchased from Sigma-Aldrich Company, commodity;

[0083]The cytotoxicity of PEI 25kDa / DNA complex and the PPP-PEP / DNA gene delivery system (ZNF580 gene) prepared in embodiment 1 and embodiment 2 is determined by MTT(3-(4,5-dimethylthiazole-2)-2 , 5-diphenyltetrazolium bromide) colorimetric method for evaluation.

[0084] The steps are as follows: the EA.hy926 cells were mixed with 1×10 4 The density of cells / well was seeded in a 96-well plate and cultured overnight with complete medium (10W% FBS DMEM). When the cells reached 80% confluency, the complete medium was replaced with serum-free medium for 12 hours. Add the PEI 25kDa / DNA complex and the different concentrations of PPP-PEP / DNA gene delivery systems prepared in Example 1 and Example 2 to the serum-free medium, and after the cells we...

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Abstract

The invention discloses a star-shaped multi-targeting functional gene vector based on POSS and application. A preparation method of the vector includes: allowing octaamino POSS and 2-bromoisobutyryl bromide to have reaction to generate macroinitiator POSS-(Br)n, performing two-step atom transfer polymerization reaction to graft hydrophilic dimethylaminoethyl methacrylate and poly(ethylene glycol)methacrylate to the POSS, introducing double bonds to the tail end of polymer, and modifying targeting functional polypeptide to the polymer to obtain the star-shaped multi-targeting functional gene vector based on POSS. A gene delivery system can be prepared by using the star-shaped multi-targeting functional gene vector based on POSS. The star-shaped multi-targeting functional gene vector based on POSS has the advantages that the star-shaped gene vector allows positive charges to be concentrated, so that the gene carrying ability of the polymer is enhanced; stable micelles small in particle size can be formed easily, and cell uptake is benefited; the PDMAEMA chain segment is used as the main chain segment for carrying the compressed gene, so that endosome escaping ability can be enhanced.

Description

technical field [0001] The invention belongs to the technical field of biological materials, and relates to a POSS-based star-shaped multi-targeting functional gene carrier and its application. Background technique [0002] In recent years, the potential application of gene therapy in multiple diseases such as malignant tumors and cardiovascular diseases has attracted more and more attention. This technology requires the introduction of exogenous normal genes and therapeutic genes into target cells to compensate for the disease caused by gene defects, thereby treating the disease. However, naked DNA cannot pass through the cell membrane, and the complex in vivo environment makes the efficiency of gene delivery very low. Therefore, people have been exploring and researching safe and efficient gene delivery systems. [0003] As a non-viral gene carrier, the star-shaped cationic polymer has a compact molecular structure and can have multiple functional groups at the end, showi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/87C12N15/62C08F283/06C08F283/12C08F220/34C08F285/00C08F220/06
CPCC12N15/87C08F283/065C08F283/122C08F285/00C07K2319/01C07K2319/10C07K2319/09C08F220/34C08F220/06
Inventor 冯亚凯王君朵兴红任相魁郭锦棠
Owner TIANJIN UNIV
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