Alicyclic compound modified high-molecular material and preparation method and application thereof

A technology of polymer materials and alicyclic compounds, which is applied in the field of polymer materials modified by alicyclic compounds and its preparation, to achieve the effects of increasing gene transfection efficiency, easy synthesis, and low cytotoxicity

Active Publication Date: 2015-12-16
EAST CHINA NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although the covalent compound formed by such alkanes and cationic polymers has high gene transfection efficiency, it still has certain cytotoxicity that cannot be ignored.

Method used

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  • Alicyclic compound modified high-molecular material and preparation method and application thereof
  • Alicyclic compound modified high-molecular material and preparation method and application thereof
  • Alicyclic compound modified high-molecular material and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The present invention is based on the gene transfection material of polymer and alicyclic compound, namely the polymer material (G5-NCO-HC modified by alicyclic compound) 5 -x,G5-NCO-HC 7 -y, G5-NCO-AD-z, G5-NCO-HC 12 -m, PEI-NCO-HC 12 -m) preparation and characterization

[0060] (1) if figure 1 Shown is the gene transfection carrier based on the polymer skeleton and the functional group of the alicyclic compound, that is, the alicyclic compound-modified polymer material G5-NCO-HC 5 The synthetic route of -x and the structural diagram of the gene transfection vector, G5-NCO-HC 5 The structural formula of -x is as shown in formula (1A):

[0061]

[0062] Wherein, R=G5, that is, the 5th generation polyamide-amine dendrimer, which has a central core and the terminal group is a primary amine group;

[0063] S=HC 5 , namely cyclopentane;

[0064] x represents the amount of cyclopentane covalently attached to the surface of the fifth generation polyamidoamine dend...

Embodiment 2

[0115] Example 2: Gene transfection vector G5-NCO-HC 5 -29 Efficiency experiment of knocking out luciferin gene in Hela-luci cells

[0116] The gene transfection carrier G5-NCO-HC that makes in embodiment 1 5 -29 and luciferase luciferase siRNA form a complex at room temperature, then transfect in Hela-luci cells, and evaluate G5-NCO-HC by detecting the expression of luciferase 5 -29 gene transfection efficiency. The specific experimental method is as follows: Hela-luci cells were incubated in a 24-well plate for 24 hours, and 50 nM luciferase siRNA was mixed with a certain amount of G5-NCO-HC 5 -29 was mixed and added to the medium and mixed evenly; after 6 hours of incubation with the cells, 500 microliters of medium containing 10% serum was added, and the cells were treated after 18 hours. For the detection method of luciferase expression, refer to the instructions provided by the manufacturer (Promega). Lipofectamine2000 was used as a positive control.

[0117] Experi...

Embodiment 3

[0119] Example 3: Gene Transfection Vector G5-NCO-HC 7 -64 Efficiency experiment of knocking out luciferin gene in Hela-luci cells

[0120] The gene transfection carrier G5-NCO-HC that makes in embodiment 1 7 -64 and luciferase luciferase siRNA form a complex at room temperature, then transfect in Hela-luci cells, and evaluate G5-NCO-HC by detecting the expression of luciferase 7 -64 gene transfection efficiency. The specific experimental method is as follows: Hela-luci cells were incubated in a 24-well plate for 24 hours, and 50 nM luciferase siRNA was mixed with a certain amount of G5-NCO-HC 7 -64 was mixed and added to the culture medium and mixed evenly; after 6 hours of incubation with the cells, 500 microliters of medium containing 10% serum was added, and the cells were treated 18 hours later. For the detection method of luciferase expression, refer to the instructions provided by the manufacturer (Promega). Lipofectamine2000 was used as a positive control.

[0121...

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Abstract

The invention provides an alicyclic compound modified high-molecular material. The alicyclic compound modified high-molecular material comprises an alicyclic compound gene and a cationic high-molecular skeleton, wherein the alicyclic compound gene is covalently connected with the surface of the cationic high-molecular skeleton, the cationic high-molecular skeleton comprises polyamidoamine dendrimers, polypropyleneimine dendrimers, polylysine dendrimers, branched polyethyleneimine cationic polymers and linear polyethyleneimine cationic polymers. The invention further provides a preparation method of the high-molecular material and application as a biological macromolecular transfection vector. The synthesis cost of the high-molecular material provided by the invention is low, the cytotoxicity is low, the biocompatibility is high, gene molecules can be effectively and safely delivered into cells, the high-molecular material can be used as the gene transfection vector having the advantages of high efficiency, low toxicity, low cost and the like, and the application prospect is good.

Description

technical field [0001] The invention relates to the technical fields of polymer chemistry, organic synthesis and biomaterials, in particular to an alicyclic compound-modified polymer material and its preparation method and application. Background technique [0002] Gene transfection refers to the process of introducing foreign genes into cells to obtain new genetic traits. Gene transfection vector is the core of gene transfection technology. An ideal gene transfection vector should have the following characteristics: high transfection efficiency, low cytotoxicity, good biological safety, and low price. Currently used gene transfection vectors mainly include viral vectors and non-viral vectors. The main application research still uses viral vectors with high transfection efficiency, but viral vectors have problems such as limited ability to carry genes and potential safety hazards. Therefore, many non-viral gene vectors have become new research hotspots, including cationic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G83/00C08G73/04C08G69/48C12N15/87
Inventor 程义云沈湾湾刘红梅
Owner EAST CHINA NORMAL UNIV
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