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REDV polypeptide modified amphiphilic comb-type gene vector and preparation method thereof

A gene carrier and polypeptide modification technology, which is applied in the field of parental comb-shaped gene carrier and preparation, can solve toxicity and other problems, and achieve the effects of improved adhesion, good target recognition performance, and low biological toxicity

Inactive Publication Date: 2016-02-10
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, high-molecular-weight polyethyleneimine is highly toxic to organisms. Therefore, a multi-site gene carrier system is designed to effectively link small-molecular-weight polyethyleneimine, which can not only ensure the effective delivery of DNA, but also reduce the Systemic toxicity is the direction of the majority of gene therapy workers

Method used

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  • REDV polypeptide modified amphiphilic comb-type gene vector and preparation method thereof
  • REDV polypeptide modified amphiphilic comb-type gene vector and preparation method thereof
  • REDV polypeptide modified amphiphilic comb-type gene vector and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1: the preparation method of the amphipathic comb-shaped gene carrier modified with REDV polypeptide may further comprise the steps:

[0058] (1) Preparation of polyhydroxyethyl methacrylate comb macroinitiator (II-1) with phenyl ring group:

[0059] Under the protection of nitrogen atmosphere, the polymerization tube was baked at high temperature to remove water, and after natural cooling, 37.0 mmol of hydroxyethyl methacrylate reaction monomer, 0.058 mmol of cuprous bromide and 0.117 mmol of 2,2 Dissolve '-bipyridine catalyst and 0.073mmol 1-bromoethylbenzene initiator in anhydrous methyl ethyl ketone solvent, after sealing, under liquid nitrogen cooling, vacuumize repeatedly, operate nitrogen gas three times, and react at 60°C for 20 hours A mixed solution was obtained, which was diluted with dichloromethane and passed through a silica gel column to remove unreacted cuprous bromide, then precipitated with glacial ether and dried in vacuo to obtain the whit...

Embodiment 2

[0088] Embodiment 2: the preparation method of the amphipathic comb-shaped gene carrier modified with REDV polypeptide may further comprise the steps:

[0089] (1) Preparation of polyhydroxyethyl methacrylate comb macroinitiator (II-2) with phenyl ring group:

[0090] Under the protection of nitrogen atmosphere, the polymerization tube was baked at high temperature to remove water, and after natural cooling, while blowing nitrogen flow, 600 mmol of hydroxyethyl methacrylate reaction monomer, 1.2 mmol of cuprous bromide and 2.4 mmol of 2,2' were sequentially added -Bipyridine catalyst and 1mmol 1-bromoethylbenzene initiator are dissolved in anhydrous methyl ethyl ketone solvent, after sealing, under liquid nitrogen cooling, vacuuming is repeated, and nitrogen gas is operated 5 times, and the mixture is reacted at 80°C for 24 hours solution, the mixture was diluted with dichloromethane and then passed through a neutral alumina column to remove unreacted cuprous bromide, then pre...

Embodiment 3

[0109] Embodiment 3: the preparation method of the amphipathic comb-shaped gene carrier modified with REDV polypeptide may further comprise the steps:

[0110] (1) Preparation of polyhydroxyethyl methacrylate comb macroinitiator (II-3) with phenyl ring group:

[0111] Under the protection of nitrogen atmosphere, the polymerization tube was baked at high temperature to remove water, and after natural cooling, while blowing nitrogen flow, 20mmol hydroxyethyl methacrylate reaction monomer, 0.1mmol cuprous bromide and 0.2mmol2,2' were sequentially added -Bipyridine catalyst and 0.1mmol 1-bromoethylbenzene initiator are dissolved in anhydrous methyl ethyl ketone solvent, after sealing, under the cooling of liquid nitrogen, vacuumize repeatedly, operate with nitrogen gas 4 times, and react at 70°C for 16 hours to obtain The mixed solution was diluted with dichloromethane and passed through a neutral alumina column to remove unreacted cuprous bromide, then precipitated with glacial e...

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Abstract

The invention discloses a REDV polypeptide modified amphiphilic comb-type gene vector and a preparation method thereof. The preparation method includes: (1) preparation of hydroxyethyl methylacrylate comb-type macroinitiator (II) with a benzene ring group; (2) preparation of a hydrophobic polyester kernel (III); (3) preparation of a carboxyl terminal hydrophobic polyester kernel (IV); (4) preparation of a comb-type polymer (V) modified by two hydrophilic segments; (5) reaction of the comb-type polymer (V) and heterodoxy polyethylene glycol modified by a pyridine disulfide group and a succinimide ester group, and addition of CREDVW for a polypeptide reaction to generate the REDV polypeptide modified amphiphilic comb-type gene vector. The REDV polypeptide modified amphiphilic comb-type gene vector is capable of efficiently loading genes, and the REDV polypeptide can be effectively combined with proteins on the surfaces of endothelial cell membranes to improve adhesion of the polymer material on the surfaces of the endothelial cell membranes, and the content of polypeptides linked on the surface of the polymer can be detected by means of fluorescence.

Description

technical field [0001] The invention relates to a parental comb-shaped gene carrier modified with REDV polypeptide and a preparation method thereof, belonging to the technical field of gene carriers with biological target recognition function. Background technique [0002] Cardiovascular disease has become one of the most important diseases threatening human life. At present, the treatment of cardiovascular diseases mainly uses biological scaffold materials to construct artificial blood vessels. However, with the effects of elastic recoil, intimal hyperplasia, and vascular remodeling, stent materials often cause restenosis and thrombosis after surgery, endangering the lives of patients. It is an effective method to treat restenosis and thrombus to achieve good rapid in situ endothelialization on the surface of implant materials. Since vascular endothelial cells have cell membrane surface membrane proteins that are different from other types of cells, using these characteri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/87A61K48/00C08G81/00
Inventor 冯亚凯王海霞郭锦棠
Owner TIANJIN UNIV
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