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Polypeptide-modified brain-targeted nanometer gene delivery system and preparation method thereof

A peptide modification, brain targeting technology, applied in gene therapy, pharmaceutical formulations, non-active components of polymer compounds, etc., to achieve the effects of increasing accumulation, prolonging circulation time, and increasing expression.

Inactive Publication Date: 2012-12-26
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are various limitations in gene delivery in the brain at present, and there is an urgent need for a new type of brain-targeted nano-gene delivery system, which can increase the uptake and crossing efficiency of the nano-gene delivery system in vitro blood-brain barrier model , to increase the accumulation of the nano-gene delivery system in the brain of animals, thereby increasing the expression of reporter genes in the brain, and realizing the release of genes across the blood-brain barrier into the brain

Method used

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  • Polypeptide-modified brain-targeted nanometer gene delivery system and preparation method thereof
  • Polypeptide-modified brain-targeted nanometer gene delivery system and preparation method thereof
  • Polypeptide-modified brain-targeted nanometer gene delivery system and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0083] Take 2mg of polyamide-amine (PAMAM, 77.35mg / ml methanol solution) and dry it in a vial, add 0.48mg polyethylene glycol (MAL-PEG3500-NHS, 1mg / ml 0.035M pH 8.0 phosphate solution), two The molar ratio is 1:2, stirred and reacted at room temperature for 2 hours, the NHS group reacts specifically with the amino group on the surface of PAMAM to prepare a polyamide-amine-polyethylene glycol (PAMAM-PEG) complex solution, MWCO 10000 ultra Unreacted polyethylene glycol (PEG3500) was removed by ultrafiltration at 12,000 rpm for 30 minutes in a centrifuge tube, and the synthesis of the carrier was characterized by proton nuclear magnetic resonance spectroscopy.

Embodiment 2

[0085] Take 2mg of polyamide-amine (PAMAM, 77.35mg / ml methanol solution) and dry it in a vial, add 0.48mg polyethylene glycol (MAL-PEG3500-NHS, 1mg / ml 0.035M pH 8.0 phosphate solution), two The molar ratio is 1:2, stirred and reacted at room temperature for 2 hours, the NHS group reacts specifically with the amino group on the surface of PAMAM to prepare a polyamide-amine-polyethylene glycol (PAMAM-PEG) complex solution, MWCO 10000 ultra Remove unreacted polyethylene glycol (PEG3500) by ultrafiltration at 12000rpm for 30min in a centrifuge tube, redissolve in pH 7.0 phosphate buffer, add 0.23mg RVG29 polypeptide (2mg / ml pH 7.0 phosphate solution), and the molar ratio of PAMAM 1:1, stirred at room temperature for 24 hours, the MAL group reacted specifically with the sulfhydryl group on the cysteine ​​residue of the RVG29 polypeptide to produce polyamide-amine-polyethylene glycol-polypeptide (PAMAM-PEG-RVG29) Complex, MWCO 10000 ultrafiltration centrifuge tube with 12000rpm ultr...

Embodiment 3

[0087] Take 2 mg of polyamide-amine (PAMAM, 77.35 mg / ml methanol solution) and dry it in a vial, and redissolve it in phosphate buffer with a pH value of 7.4 to prepare a carrier solution. Dissolve the plasmid DNA in 50mM sodium sulfate solution, dilute it to 0.1mg / ml, make the molar ratio of polyamide-amine to 1:10, vortex at room temperature for 30s, and make non-targeting PAMAM / DNA loaded with genes nanoparticles.

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Abstract

The invention belongs to the field of biotechnology and relates to a street virus glycoprotein derivative polypeptide-modified brain-targeted nanometer gene delivery system and a preparation method thereof. The preparation method comprises the following steps of: connecting a hydrophilic polymer with an RVG29 polypeptide to construct a brain-target carrier by adopting a street virus glycoprotein derivative peptide RVG29 as a brain-targeted head group and cationic dendritic polymers as carrier materials; and compounding the brain-target carrier and a plasmid deoxyribonucleic acid (DNA) for coding a specific gene through an electrostatic compound effect to prepare the brain-targeted nanometer gene delivery system. The intake and penetration efficiency of the nanometer gene delivery system on an in-vitro blood brain barrier model can be increased; the accumulation of the nanometer gene delivery system in an animal brain is increased; the expression quantity of a reporter gene in the brain is increased; the gene is delivered into the brain when crossing a blood brain barrier; and a high-efficiency safe strategy is provided for the application of diagnosing and treating gene in the brain.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a polypeptide-modified brain-targeted nano-gene delivery system, in particular to a rabies virus glycoprotein-derived polypeptide-modified brain-targeted nano-gene delivery system and a preparation method thereof. Background technique [0002] Studies have shown that the existence of the blood-brain barrier plays a vital role in maintaining the homeostasis of the central nervous system, but at the same time, it also hinders the effective delivery of diagnostic or therapeutic drugs into the brain. Modern medical research has confirmed that the main component of the blood-brain barrier is brain capillary endothelial cells, which are characterized by dense tight junctions between cells, and it is difficult for most substances to pass through; It can cross the blood-brain barrier and reach the brain by binding to specific receptors and using receptor-mediated penetrating effects; i...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K47/42A61K47/34
Inventor 蒋晨刘洋
Owner FUDAN UNIV
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