PEGylated Fe3+/ PEI genetic vector based on functional peptide R9 modification and preparation method and application of PEGylated Fe3+/ PEI genetic vector

A gene carrier and functional peptide technology, applied in the medical field, can solve the problems of poor targeting of polyethyleneimine, strong cytotoxicity, and easy dissociation, so as to improve nuclear delivery capacity, reduce cytotoxicity, and improve transfection efficiency. Effect

Active Publication Date: 2017-03-08
SHANGHAI OCEAN UNIV
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are two outstanding problems in the use of polyethyleneimine 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 at physiological ion concentration, resulting in poor transfection effect; although PEI with a molecular weight above 20kd has a relatively ideal transfection efficiency, but because th

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • PEGylated Fe3+/ PEI genetic vector based on functional peptide R9 modification and preparation method and application of PEGylated Fe3+/ PEI genetic vector
  • PEGylated Fe3+/ PEI genetic vector based on functional peptide R9 modification and preparation method and application of PEGylated Fe3+/ PEI genetic vector
  • PEGylated Fe3+/ PEI genetic vector based on functional peptide R9 modification and preparation method and application of PEGylated Fe3+/ PEI genetic vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 PEG-Fe modified by polypeptide R9 3+ Preparation and functional verification of / PEI gene carrier (1)

[0045] 1. PEG-Fe 3+ Synthesis of / PEI

[0046] 20 μL FeCl 3 ·6H 2 O (250mg / mL), 30μL bPEI (250mg / mL), 10μL vinyl sulfone (10mg / mL), 15μL PEG-NHS (Mw=5000, 10mg / mL) were sequentially added to 1mL Tween 80 cyclohexane solution (250mg / mL mL), room temperature, fully stirred for 12h. After the reaction, add excess ethanol to the emulsion, centrifuge at 10,000r / min for 1 hour, remove the supernatant carefully, and disperse the precipitate in ethanol again by sonication. Pure water dissolves the precipitate, and the result is PEG-Fe 3+ / PEI polymer solution.

[0047] 2. Synthesis of functional peptide R9

[0048] The sequence of the functional peptide R9 is Arg-Gly-Asp-Cys-Lys-Lys-Lys-Arg-Lys, which was synthesized by Shanghai Gil Biochemical Co., Ltd. by solid-phase method.

[0049] 3. PEG-Fe modified by polypeptide R9 3+ Preparation of / PEI

[0050] S...

Embodiment 2

[0055] Example 2 PEG-Fe modified by polypeptide R9 3+ Preparation and functional verification of / PEI gene carrier (2)

[0056] PEG-Fe 3+ The preparation, cytotoxicity test, and in vitro transfection experiment of / PEI-R9 are the same as in Example 1, except that the PEG-Fe prepared in this example 3+ / PEI-R9, where PEI and Fe 3+ The molar ratio of PEI is 1:6, the molecular weight of PEI is 5KDa, and the molar ratio of PEGylated FE3+ / PEI to polypeptide R9 is 1:5.

[0057] Cytotoxicity test results showed that: PEG-Fe 3+ / PEI-R9 is almost non-toxic, the PEG-Fe of the present embodiment 3+ / PEI-R9 has low cytotoxicity, ensuring that PEG-Fe 3+ / The feasibility of PEI-R9 as a gene carrier. The results of in vitro transfection experiments are shown in Table 1.

Embodiment 3

[0058] Example 3 PEG-Fe modified by polypeptide R9 3+ Preparation and functional verification of / PEI gene carrier (3)

[0059] PEG-Fe 3+ The preparation, cytotoxicity test, and in vitro transfection experiment of / PEI-R9 are the same as in Example 1, except that the PEG-Fe prepared in this example 3+ / PEI-R9, where PEI and Fe 3+ The molar ratio of the PEGylated FE3+ / PEI to the polypeptide R9 is 1:10, the molecular weight of PEI is 25KDa, and the molar ratio of the PEGylated FE3+ / PEI to the polypeptide R9 is 1:10.

[0060] Cytotoxicity test results showed that: PEG-Fe 3+ / PEI-R9 is almost non-toxic, the PEG-Fe of the present embodiment 3+ / PEI-R9 has low cytotoxicity, ensuring that PEG-Fe 3+ / The feasibility of PEI-R9 as a gene carrier. The results of in vitro transfection experiments are shown in Table 1.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a PEGylated Fe3+/ PEI genetic vector based on functional peptide R9 modification and a preparation method and application of the PEGylated Fe3+/ PEI genetic vector. The genetic vector contains polyethylene glycol modified Fe3+/ polyethyleneimine and a conjugate formed by functional peptides R9. The PEGylated Fe3+/ PEI genetic vector based on functional peptide R9 modification has the advantages that metal ions Fe3 react with PEI at first, PEG modification is carried out to form PEGylated Fe3+/ PEI, then the functional peptide R9 which has ability of improving nuclear delivery and is targeted at alpha v beta 3 is synthesized, R9 is coupled with a PEG-Fe3+/ PEI by a crosslinking technology, and therefore, a novel virogene-free vector system PEG-Fe3 +/ PEI is established. On the basis of reducing PEI cytotoxicity, in-vivo targeting property and nuclear delivery ability are improved, then the transfection efficiency of the vector in vivo is improved, and an effective way is searched for final application of gene therapy on clinical treatment.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to a PEGylated Fe modified based on functional peptide R9 3+ / PEI gene carrier and its preparation method and application. Background technique [0002] Gene therapy is a new treatment method based on genetic engineering technology and molecular genetics in recent years. Since the biological basis of tumor occurrence and development is gene mutation, gene therapy has become the most promising field for overcoming tumors. There are three important links in gene therapy, namely the target gene, transgenic carrier and target cells. The gene transfer system is the core technology of gene therapy. The biggest problem at this stage is that the ideal gene carrier has not been found yet. Currently applied vectors include two categories: viral vectors and non-viral vectors. Viral vectors have high transfection efficiency but have problems such as low carrying capacity and potential safet...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/85C12N15/64C07K7/06A61K47/42A61K48/00A61K31/713A61P35/00
Inventor 刘克海张敏邬军文余倩张庭瑞吴文惠
Owner SHANGHAI OCEAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap