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Modified polyethyleneimine, a gene vector composition, and a preparing method and applications of the gene vector composition

A technology of polyethylenimine and gene carrier, which is applied in the direction of introducing foreign genetic material by using carrier, other methods of inserting foreign genetic material, gene therapy, etc. It can solve the problems of low transfection efficiency, no active targeting of tumor cells, strong cytotoxicity

Inactive Publication Date: 2016-07-06
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the transfection efficiency of PEI is related to its molecular weight. Although PEI with a molecular weight above 20KDa has a strong transfection process, it also has strong cytotoxicity, while PEI with a small molecular weight (2000) has low cytotoxicity, but Its transfection efficiency is low
In the prior art, when the derivatives obtained by chemically modifying PEI are used as gene carriers, they still do not have active targeting to tumor cells, and their stability and biodegradability are poor.

Method used

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  • Modified polyethyleneimine, a gene vector composition, and a preparing method and applications of the gene vector composition
  • Modified polyethyleneimine, a gene vector composition, and a preparing method and applications of the gene vector composition
  • Modified polyethyleneimine, a gene vector composition, and a preparing method and applications of the gene vector composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 14

[0071] Preparation of polyethyleneimine (PEI-PBA) modified by embodiment 14-carboxyphenylboronic acid

[0072] (1) Weigh 100 mg of 4-carboxyphenylboronic acid (abbreviated as PBA), dissolve it in 10 mL of methanol solvent, add 100 mg of N-hydroxysuccinimide (abbreviated as NHS), stir magnetically at room temperature, and then add 280 mg 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (abbreviated as EDC), activated at room temperature for 4 hours to obtain activated 4-carboxyphenylboronic acid;

[0073] (2) Take a 100mL reaction bottle, add 300mg of polyethyleneimine (Mw=1800, abbreviated as PEI 1.8K ) and 10mL of methanol, then add the activated 4-carboxyphenylboronic acid into the reaction flask, and react at 25°C for 24 hours;

[0074] (3) After the reaction is over, add 200 mL of ether into the reaction flask described in step (2), repeat this twice to obtain a precipitate, collect the precipitate, separate the precipitate, and dry it in vacuum to obtain the c...

Embodiment 2

[0080] Preparation and characterization of embodiment 2PEI-PBA gene carrier complex

[0081] (1) Weigh 10 mg of the gene carrier and dissolve it in 10 mL of double-distilled water to obtain a gene carrier solution (concentration is 1 mg / mL). The gene carrier is the polyethyleneimine derivative PEI-PBA prepared in Example 1;

[0082] (2) Take 6.4 mL of the gene carrier solution described in step (1), add 1 μg RNA, vortex mix for 5 min, and incubate at room temperature for 20 min to obtain the PEI-PBA gene carrier complex.

[0083] figure 2 The particle size diagram of the gene-carrier complex formed after the 4-carboxyphenylboronic acid modified PEI-PBA loaded with the gene prepared in 2, the particle size is measured by dynamic light scattering on the Malvern nano particle size analyzer ,From figure 2 It can be seen that the particle size of the PEI-PBA gene carrier complex is relatively uniform, with an average particle size of 156 nm.

[0084] The value of Zeta potentia...

Embodiment 3

[0085] The preparation of embodiment 3PEI-PBA gene carrier complex

[0086]The PEI-PBA gene carrier complex was prepared according to the technical scheme in Example 2 of the present invention, and the PEI-PBA used was prepared in Example 1 of the present invention. The difference is that the genetic material carried by PEI-PBA in this example is Green fluorescent protein particle (pEGFP-N1), please refer to Table 1 for details.

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Abstract

The invention provides modified polyethyleneimine, and a preparing method and applications of the modified polyethyleneimine. The modified polyethyleneimine is prepared through modifying polyethyleneimine with different molecular weights with phenylboronic acid compounds. The modified polyethyleneimine has an active targeting property for tumor cell membranes, and can carry genes to form a gene vector composition used for gene therapy. The gene vector composition is uniform in particle size, proper in charge, stable in properties, low in cell toxicity, high in cell transfection efficiency, high in target power, and good in biocompatibility and biodegradability. The modified polyethyleneimine and the gene vector composition can be used for the gene therapy. Preparing processes of the modified polyethyleneimine and the gene vector composition are simple, high in reaction degree controllability and capable of industrial production.

Description

technical field [0001] The invention relates to the technical field of biological carriers, in particular to a modified polyethylenimine, a gene carrier complex and a preparation method and application thereof. Background technique [0002] Gene therapy achieves the purpose of treating diseases by introducing exogenous genes into the nucleus of target cells to repair defective genes that cause diseases or inhibit harmful genes that cause diseases. Successful gene therapy depends on effective gene carriers, and common carriers are divided into viral vectors and non-viral vectors. The development of viral vectors is limited due to the great safety hazards; non-viral vectors are mostly cationic polymers, which are easy to prepare and transport, safe and effective, and have no immunogenicity, etc., and have become a research hotspot for gene vectors one. [0003] Polyethyleneimine (referred to as PEI) is a type of non-viral carrier widely used at present. It is rich in cations...

Claims

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Application Information

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IPC IPC(8): C08G73/04C12N15/87C12N15/85A61K48/00A61K47/32
Inventor 马轶凡姬满祎李萍盛楠
Owner SHENZHEN INST OF ADVANCED TECH
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