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Gene vector modified by bifunctional peptide and preparation method thereof

A bifunctional peptide and gene carrier technology, applied in the field of gene carrier, to achieve the effect of enhancing targeting, reducing cell toxicity, and reducing cytotoxicity

Inactive Publication Date: 2012-04-04
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still two outstanding problems in the use of PEI as a gene carrier: first, there is a contradiction between transfection efficiency and cytotoxicity
Although small molecule PEI has low cytotoxicity, it is easy to dissociate with DNA under physiological ion concentration, resulting in poor transfection effect; although PEI with a molecular weight above 20 kDa has a relatively ideal transfection efficiency, because the surface of PEI is rich in Positive charge and non-degradability in vivo, resulting in high molecular weight PEI showing strong cytotoxicity
Second, polyethyleneimine has poor targeting
But there is no report about the Pluronic P123-polyethyleneimine modified by bifunctional peptides and its preparation method and application

Method used

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  • Gene vector modified by bifunctional peptide and preparation method thereof
  • Gene vector modified by bifunctional peptide and preparation method thereof
  • Gene vector modified by bifunctional peptide and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] 1. Preparation of P123-PEI

[0049] Weigh 0.6 mmol of P123 after water removal, dissolve it in 40 ml, and add 1.2 mmol of bis(tri Chloromethyl) carbonate, 180rpm magnetic stirring reaction at room temperature overnight. Remove the solvent by rotary evaporation in vacuo, then dissolve with 30ml of anhydrous toluene and anhydrous dichloromethane mixed solution with a volume ratio of anhydrous toluene and anhydrous dichloromethane of 2:1, and then add 2.0mmol N-hydroxysuccinimide , under 180rpm magnetic stirring, 2.0mmol anhydrous triethylamine was added dropwise into the reaction solution, and the stirring reaction was continued for about 4h. After the reaction is complete, the reaction solution is filtered and the solvent is removed by vacuum rotary evaporation again. The obtained residue is dissolved in 50ml of ethyl acetate, centrifuged at 8000rpm for 15min, and the supernatant is taken, rotary evaporated, and the ethyl acetate is evaporated, and the reactant is coole...

Embodiment 2

[0059] 1. Preparation of P123-PEI

[0060] Weigh 0.6mmol of P123 after water removal, dissolve it in 40ml, and add 0.6mmol of bis(tri Chloromethyl) carbonate, 180rpm magnetic stirring reaction at room temperature overnight. Remove the solvent by rotary evaporation in vacuo, then dissolve with 30ml of anhydrous toluene and anhydrous dichloromethane mixed solution with a volume ratio of anhydrous toluene and anhydrous dichloromethane of 2:1, and then add 2.0mmol N-hydroxysuccinimide , under 180rpm magnetic stirring, 2.0mmol anhydrous triethylamine was added dropwise into the reaction solution, and the stirring reaction was continued for about 4h. After the reaction is complete, the reaction solution is filtered and the solvent is removed by vacuum rotary evaporation again. The obtained residue is dissolved in 50ml of ethyl acetate, centrifuged at 8000rpm for 15min, and the supernatant is taken, rotary evaporated, and the ethyl acetate is evaporated, and the reactant is cooled a...

Embodiment 3

[0075] 1. Preparation of P123-PEI

[0076] Weigh 0.6mmol of P123 after water removal, dissolve it in 40ml, and add 12mmol of bis(trichloromethane Methyl) carbonate, 180rpm magnetic stirring reaction at room temperature overnight. Remove the solvent by rotary evaporation in vacuo, then dissolve with 30ml of anhydrous toluene and anhydrous dichloromethane mixed solution with a volume ratio of anhydrous toluene and anhydrous dichloromethane of 2:1, and then add 2.0mmol N-hydroxysuccinimide , under 180rpm magnetic stirring, 2.0mmol anhydrous triethylamine was added dropwise into the reaction solution, and the stirring reaction was continued for about 4h. After the reaction is complete, the reaction solution is filtered and the solvent is removed by vacuum rotary evaporation again. The obtained residue is dissolved in 50ml of ethyl acetate, centrifuged at 8000rpm for 15min, and the supernatant is taken, rotary evaporated, and the ethyl acetate is evaporated, and the reactant is co...

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Abstract

The invention relates to a gene vector modified by a bifunctional peptide and a preparation method and application thereof. An amino acid sequence of the bifunctional peptide is shown as SEQIDNO.1. The gene vector is formed in a mode that the bifunctional peptide is coupled with polyethyleneimine derivatives. The polyethyleneimine derivatives are polyethyleneimine modified by planck P123. The invention also relates to a preparation method of the gene vector and application of the bifunctional peptide and the gene vector to the gene therapy. The bifunctional peptide provided by the invention has strong targeting and membrane penetrating capability. The gene vector provided by the invention has high cell transfection efficiency and low cytotoxicity. An effective means is provided for the gene therapy of diseases.

Description

technical field [0001] The invention relates to the technical field of gene carriers, in particular to a gene carrier modified with bifunctional peptides and a preparation method thereof. Background technique [0002] Gene therapy is a new treatment method based on genetic engineering technology and molecular genetics in recent years. Because the biological basis of tumor occurrence and development is gene mutation, gene therapy has become the most promising means to overcome tumors. the most active areas of research. There are three important links in gene therapy, namely target gene, transgenic carrier and target cells, and its core technology is the establishment of gene transfer system. The biggest problem facing gene therapy at this stage is that the ideal gene carrier has not yet been found. Currently applied vectors can be divided into two categories: viral vectors and non-viral vectors. Viral vectors have high transfection efficiency but have disadvantages such a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C12N15/63C08G81/00
Inventor 刘克海朱青高申王晓宇
Owner SHANGHAI OCEAN UNIV
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