Chlorotoxin-modified glioma targeting gene delivery compound and preparation method thereof

A chlorinated toxin and brain glioma technology, applied in gene therapy, medical preparations of non-active ingredients, non-active ingredients of polymer compounds, etc., can solve problems such as strong hemolysis, increase expression and increase transfection Efficiency and expression level, effect of increasing transfection efficiency and expression level

Inactive Publication Date: 2012-05-23
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the use of PAMAM alone has certain shortcomings, such as strong hemolys...

Method used

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  • Chlorotoxin-modified glioma targeting gene delivery compound and preparation method thereof
  • Chlorotoxin-modified glioma targeting gene delivery compound and preparation method thereof
  • Chlorotoxin-modified glioma targeting gene delivery compound and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] PAMAM and PEG containing bifunctional groups were dissolved in phosphate buffer saline (PBS for short) at pH 8.0 according to the molecular ratio of 1:2, stirred and reacted at room temperature for 120 minutes to generate PAMAM-PEG, unreacted PEG was removed by ultrafiltration and replaced The buffer was PBS pH 7.0. At the same time, chlorinated toxins were reacted with Traut's thiol reagent to introduce thiol groups, and after purification, they were mixed with PAMAM-PEG at a molecular ratio of 1:1, stirred and reacted at room temperature for 24 hours to generate PAMAM-PEG-CTX, and molecular exclusion chromatography PAMAM-PEG-CTX can be obtained by removing the unreacted chlorinated toxin.

Embodiment 2

[0077] PAMAM and PEG containing bifunctional groups were dissolved in PBS at pH 8.0 according to the molecular ratio of 1:6, stirred at room temperature for 120 minutes to generate PAMAM-PEG, unreacted PEG was removed by ultrafiltration and the buffer was replaced with PBS at pH 7.0 . At the same time, the chlorinated toxin was reacted with Traut's thiol reagent to introduce sulfhydryl groups, and after purification, it was mixed with PAMAM-PEG at a molecular ratio of 3:1, stirred and reacted at room temperature for 24 hours to generate PAMAM-PEG-CTX, and molecular exclusion chromatography PAMAM-PEG-CTX can be obtained by removing the unreacted chlorinated toxin.

Embodiment 3

[0079] The PAMAM-PEG-CTX gene delivery carrier prepared in Example 1 was dissolved in an appropriate amount of PBS with a pH of 7.4 to form a solution of 300 mg / ml (calculated based on the mass of PAMAM), and the pEGFP-N2 green fluorescent protein therapeutic gene plasmid DNA It was dissolved in an appropriate amount of 50 mM sodium sulfate solution to prepare a 100 mg / ml solution, and the two were vortexed in equal volume for 30 s at room temperature to prepare the PAMAM-PEG-CTX / DNA gene delivery complex.

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Abstract

The invention belongs to the technical field of biology, and relates to a chlorotoxin-modified glioma targeting gene delivery compound and a preparation method thereof. In the invention, chlorotoxin is modified onto a cationic high polymer material through a hydrophilic polymer, and is compounded with a plasmid DNA (Deoxyribonucleic Acid) under an electrostatic action, so that a gene delivery compound is formed. Due to the adoption of the gene delivery compound, the transfection efficiency and expression level of a gene in a glioma cell can be effectively increased; and in particular, after administration is performed in a non-invasive intravenous injection way, so that the gene expression of the gene delivery compound in a glioma lesion location is remarkably improved. The chlorotoxin-modified glioma targeting gene delivery compound in the invention can be used for transfecting endothelial cells, epithelial cells, various tissue parenchyma cells and/or nerve cells derived from human bodies and animals.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a brain glioma targeting gene delivery complex, in particular to a chlorotoxin-modified brain glioma targeting gene delivery complex and a preparation method thereof. Background technique [0002] Glioma is the brain tumor with the highest incidence rate and one of the top 10 types of tumors with the highest mortality rate. Due to the invasive growth characteristics of glioma itself and the existence of the blood-brain barrier, it is difficult to achieve a good therapeutic effect with the commonly used treatment methods including surgical resection, radiotherapy, and chemotherapy. Gene therapy is a treatment with great potential, but it is difficult for gene drugs to independently enrich glioma, and it is necessary to construct a gene delivery carrier with active targeting properties, so that exogenous gene drugs can be widely expressed in the lesion. [0003] Cationic polymer...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K47/48A61K47/34A61K47/42A61P35/00
Inventor 黄容琴蒋晨柯伟伦
Owner FUDAN UNIV
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