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A kind of metal coordination polycation gene carrier and its preparation method and application

A technology of polycation and gene carrier, which is applied in the field of metal coordination polycation gene carrier and its preparation, can solve the problem of low transfection efficiency of stem cells, achieve the effects of expanding application, improving endocytic efficiency, and enhancing DNA binding ability

Active Publication Date: 2019-02-01
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the transfection efficiency of these polycations, especially for stem cells, is very low

Method used

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  • A kind of metal coordination polycation gene carrier and its preparation method and application
  • A kind of metal coordination polycation gene carrier and its preparation method and application
  • A kind of metal coordination polycation gene carrier and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A metal-coordinated polycationic gene carrier is composed of a coordinating metal, a coordinating unit, a stimulus-responsive unit and a polycation, wherein the coordinating metal is zinc (Zn), and the coordinating unit is a lutidine amine group, The stimuli-responsive unit is N,N'-bis(acryloyl)cystamine (CBA), a monomer containing disulfide bonds; the polycation is polyethyleneimine (PEI) with a weight average molecular weight of 1800, and the zinc coordination monomer modifies the polycation The structural formula of Zn-PD obtained by PEI is shown below:

[0026]

[0027] In the structural formula: "M" represents the coordination metal Zn; "n" represents the number of metal ion ligands grafted to one polyethyleneimine (PEI) molecule, and n is 2 or 4.

[0028] A method for preparing the metal-coordinated polycationic gene carrier Zn-PD, comprising the steps of:

[0029] 1) Preparation of stimuli-responsive ligand DDAC

[0030] Dissolve 16mmol of 1,4-p-dichlorobenz...

Embodiment 2

[0035]A metal-coordinated polycationic gene carrier, comprising a coordinating metal, a coordinating unit, a stimulus response unit and a polycation, wherein the coordinating metal is magnesium (Mg), and the coordinating unit is a lutidine amine group, The stimuli-responsive unit is bis(2-propene)ethoxydisulfide (DSDA), a monomer containing disulfide bonds; the polycation is the first-generation polyamide-amine (PAMAM), and the magnesium coordination monomer modifies the polycationic PAMAM to obtain The structural formula of Mg-PAD is shown below:

[0036]

[0037] In the structural formula: "M" represents the coordination metal Mg; "n" represents the number of metal ion ligands grafted to one polyamide-amine (PAMAM) molecule, and n is 1 or 3.

[0038] A preparation method of the metal-coordinated polycationic gene carrier Mg-PAD, comprising the steps of:

[0039] 1) Preparation of stimuli-responsive ligand DDAD

[0040] Dissolve 16mmol of 1,4-p-dichlorobenzyl (Dx) in 20m...

Embodiment 3

[0044] The application of the prepared metal-coordinated polycationic gene carrier is used for non-viral gene carrier to deliver deoxyribonucleic acid (DNA) into cells, the method is as follows: the metal-coordination polycation Zn-PD and DNA are mixed in equal volume, and the room temperature Let stand for 10-30 minutes to obtain a Zn-PD / DNA complex with a mass ratio of 5-60:1; the prepared complex is not only used to transfect conventional immortal cell lines, but also limited proliferation functional Cell-stem cells and with very low cytotoxicity, as follows:

[0045] Cell transfection steps for Zn-PD2 and Zn-PD4

[0046] 1) Preparation of Zn-PD / DNA complexes:

[0047] The DNAs used were green fluorescent protein plasmid pCMV-GFP and luciferase plasmid pCMV-GLuc. Dissolve Zn-PD2 or Zn-PD4 and DNA in acetate buffer at pH 5.2, mix with equal volume, and let stand for 15 minutes at room temperature to obtain a complex with a mass ratio of Zn-PD / DNA of 5-20:1 ;

[0048] 2) ...

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Abstract

The invention discloses a metal coordination polycation gene vector. The metal coordination polycation gene vector consists of a coordinated metal, a coordinated unit, a stimuli-responsive unit and polycations, and the structural general formula of the metal coordination polycation gene vector is as shown in the specification, wherein M represents the coordinated metal; a dimethyl pyridylamine component represents the coordinated unit; a disulfide bond component represents the stimuli-responsive unit; a ball represents the polycations; n represents the quantity of metal ion ligands grafted by one polycation molecule, n is 1 to 200; and the metal coordination polycation gene vector is used for non-virus gene vectors. The metal coordination polycation gene vector has the advantage that the metal ion ligands containing stimuli-responsive radicals are modified on the polycations, so that the DNA binding capacity of the polycations and the acting force with cytomembranes are greatly improved; after the compound enters cells, the metal ion ligands and the polycations are disconnected, DNAs are released and expressed so as to show extremely high transfection efficiency, so that the application of the polycations in the aspect of the non-virus gene vectors is greatly expanded due to metal coordination monomers in the invention.

Description

technical field [0001] The invention belongs to the field of gene therapy and new materials, and particularly relates to a metal-coordinated polycationic gene carrier and a preparation method and application thereof. Background technique [0002] Gene therapy refers to the process of delivering functional nucleic acid molecules to specific tissue cells by certain means for disease treatment. It can be used for the treatment of many congenital genetic diseases and severe acquired diseases. However, the lack of efficient gene carriers has become the main obstacle to the development of gene therapy, especially the transfection efficiency of some functional cells (such as stem cells) is particularly low, which seriously hinders the development of gene regulation. Stem cells are a kind of cell population with self-renewal and differentiation ability, and have attracted much attention in the fields of tissue engineering and cell therapy. Through gene regulation, stem cells can di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G73/02C08G69/48C08G83/00C12N15/87
CPCC08G69/48C08G73/0213C08G83/004C12N15/87
Inventor 郭天瑛刘帅
Owner NANKAI UNIV
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