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Chitosan nanoparticle composite as well as preparation method and application thereof

A technology of chitosan nanoparticles and complexes, which is applied in drug combinations, pharmaceutical formulations, gene therapy, etc., can solve problems such as no gene therapy drugs, achieve improved bioavailability, high biocompatibility, and solve precipitation problems Effect

Inactive Publication Date: 2012-09-19
杨建安 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In recent years, the understanding of PDGF-B has been deepening, and its mechanism and effect in the excessive proliferation of vascular smooth muscle cells have attracted much attention, but there are no specific and effective gene therapy drugs related to it.

Method used

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  • Chitosan nanoparticle composite as well as preparation method and application thereof
  • Chitosan nanoparticle composite as well as preparation method and application thereof
  • Chitosan nanoparticle composite as well as preparation method and application thereof

Examples

Experimental program
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Embodiment 1

[0036] The preparation of embodiment 1 chitosan nanoparticle complex of the present invention

[0037] 1.1 Construction of PDGF-B siRNA expression vector: two PDGF-B siRNA primers were designed online according to BD Biosciences software:

[0038] 5'—GATCCGCGAGGAGCTTTATGAGATGTCAAGAGACATCTCATAAAGCTCCTCGTTTTTTACGCGTG--3';

[0039] 5'—AATTCACGCGTAAAAAACGAGGAGCTTTATGAGATGTCTCTTGAACATCTCATAAAGCTCCTCGCG--3';

[0040] It was synthesized as double-stranded DNA (dsDNA) and ligated with the linear plasmid PSIREN-Shuttle Vectors. The expression vector was amplified, the plasmid was extracted, the enzyme digestion identification and sequencing results all met the design requirements.

[0041] 1.2 Preparation and characterization analysis of chitosan nano PDGF-B siRNA expression vector

[0042] Prepare chitosan acetic acid solution and slowly drop into acetone under the condition of magnetic stirring; homogenize under the condition of 13500rpm to obtain chitosan nanoparticle suspension;...

Embodiment 2

[0043] Embodiment 2 is to the characterization analysis and effect analysis of the chitosan nanoparticle complex that embodiment 1 makes;

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Abstract

The invention relates to the field of gene therapy biotechnologies, and in particular relates to a chitosan nanoparticle composite capable of treating vascular injury restenosis and vascular smooth muscle proliferation as well as a preparation method, a drug and application thereof. The technical scheme adopted by the invention aims to provide the chitosan nanoparticle composite comprising small interfering (si) RNA (Ribose Nucleic Acid) aiming at PDGF (Platelet Derived Growth Factor)-B mRNA (messenger RNA) and a chitosan nanoparticle carrier thereof. According to the invention, the si RNA with specific inhibition of the PDGF-B mRNA expression is designed and synthetized on the base of a c-sis gene and the nanometer carrier is prepared by using chitosan as a raw material to ensure that the cell proliferation and the intercellular substance forming are inhibited through the local transfection of the smooth muscle cell or other relevant target cells. The chitosan nanoparticle composite gene of the PDGF-B si RNA is better in transfection effect and capable of inhibiting the occurrence and the development of relevant diseases of smooth muscle proliferation so as to achieve the treatment purpose.

Description

technical field [0001] The invention relates to the field of gene therapy biotechnology, in particular to a chitosan nanoparticle complex and a preparation method and application thereof. Background technique [0002] Growth factors are proteinaceous substances that have biological effects such as inducing and stimulating cell proliferation and maintaining cell survival. They are of great significance in promoting cell proliferation, repair and regeneration of tissues or organs, and play an important role in tissue engineering. However, all kinds of growth factors are polypeptide protein substances, which are easy to lose their biological activity in the presence of water and at room temperature. If it is directly injected into the body, the growth factor will be rapidly diffused and degraded within a day, and cannot play a long-term role in promoting the recovery of defective tissue. Therefore, how to maintain and prolong the biological activity of growth factors in the bo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K47/36A61P9/00
Inventor 杨建安季军何霞
Owner 杨建安
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