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Environment-responsive peptide-containing dendrimer lipid material and its preparation method and application

An environment-responsive, dendrimer technology, applied in the field of biomedical materials, can solve the problems of over-packaging of genetic material, high cytotoxicity and poor effect.

Active Publication Date: 2018-07-13
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This kind of high-density positive charge carrier and gene assembly is conducive to cell uptake, but it will bring side effects of high cytotoxicity, and at the same time, it will cause the genetic material to be too tightly packed and released incompletely in the target tissue or cell, resulting in positive Consequences of poor transfection of charge carriers in vitro and in vivo

Method used

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  • Environment-responsive peptide-containing dendrimer lipid material and its preparation method and application
  • Environment-responsive peptide-containing dendrimer lipid material and its preparation method and application
  • Environment-responsive peptide-containing dendrimer lipid material and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] (1) Use divergence method or convergence method or divergence-convergence method to synthesize more than two generations of dendrimers (refer to the method described in the patent literature with publication number CN 103554923A), or use commercially available dendrimers molecular;

[0071] (2) Carry out end group modification on the periphery of the dendritic molecule, and graft a large number of amino groups and / or guanidine groups on the periphery. When lysine or arginine or histidine is selected as the repeating unit of the dendrimer, the above-mentioned end group modification step is not required;

[0072] (3) Protect the peripheral functional groups of the resulting dendrimers containing amino groups and / or guanidino groups, and only expose one or two functional groups at one end, and then pass a reduction-sensitive disulfide bond or diselenide on the functional groups The bond connects the hydrophobic group;

[0073] (4) Removing the protecting groups of the am...

Embodiment 2

[0079] Gene carrier preparation scheme 1:

[0080] (1) Dissolve plasmid DNA or RNA in sterile HBG buffer solution (20 mmol of 4-hydroxyethylpiperazineethanesulfonic acid, 5% glucose) to prepare a 0.1 mg / mL gene solution (denoted as solution A); the environment-responsive lipid material prepared in Example 1 (preferably the lipid material shown in Formula 1) was added to the HBG buffer solution by ethanol injection, and prepared into a 0.1-10 mg / mL self-assembly solution , solution B;

[0081] (2) Mix the solution B obtained in the above steps with the gene solution, and incubate at room temperature for 20 minutes to obtain a binary complex, in which the gene is bound to the periphery of the self-assembly through electrostatic adsorption. The self-assembled body acts as a cationic carrier for compressing genes, and further, a shielding system can be introduced into the periphery of the above-mentioned binary complex through charge adsorption to form a ternary complex.

preparation approach 2

[0083] Dissolve the plasmid DNA in a sterile HBG buffer solution to prepare a 0.1 mg / mL DNA solution (solution A); use at least one environment-responsive peptide-containing dendrimer lipid material prepared in Example 1 The ethanol injection method or the thin-film ultrasonic method were respectively added into the solution A, and self-assembled to obtain a carrier system for efficiently assembling genes.

[0084] As an option, in the above two schemes, the pH-responsive groups containing type A and pH-responsive groups containing type B can be mixed for the preparation of gene carriers, and further, the ratio of the number of groups A to type B is 0.1 :1~10~1.

[0085] As an option, in the above two schemes, the mass ratio of the lipid material to the gene is 0.1:1-50:1.

[0086] As an option, drugs or magnetic nanoparticles can also be added in the above step (2) to prepare ternary or quaternary compounds.

[0087] Experiments have shown that the gene carriers obtained by...

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Abstract

The invention discloses an environmental response type peptide-containing dendrimer lipid material and a preparation method and application thereof, and belongs to the field of biological materials. A main body of the lipid material is dendrimer with amino and / or guanidine at the periphery, so that the hydrophilic end of the lipid material is formed; one end of the dendrimer is connected with a hydrophobic group, so that the hydrophobic end of the lipid material is formed; a connection molecule between the hydrophilic end and the hydrophobic end contains a reduction-sensitive disulfide bond or di-selenium bond; the amphipathic lipid material can be self-assembled into a lipid body or micelle or vesica with a nano size under the hydrophobic and hydrophobic effects; a medicament or gene carrier system can be formed by wrapping a medicament or a gene in the lipid body or micelle or vesica; when the carrier system enters a high-concentration reducing matter environment, the reduction-sensitive bond is broken, and disassembling is caused; the medicament or the gene can be released.

Description

technical field [0001] The invention relates to the field of biomedical materials, in particular to an assembly with environmental responsiveness and its preparation method and application. technical background [0002] More and more gene-related diseases are considered to be curable by gene therapy, which usually requires the delivery of nucleic acid drugs to target cells to treat some tumors, asthma and cardiovascular diseases. However, genes are negatively charged genetic material, and it is difficult to pass through the cell membrane by themselves. Therefore, the synthetic carriers that can effectively load them are usually high-density positively charged, and many cationic lipids and polymer systems have been developed. This kind of high-density positive charge carrier and gene assembly is conducive to cell uptake, but it will bring side effects of high cytotoxicity, and at the same time, it will cause the genetic material to be too tightly packed and released incomplet...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K47/42A61K45/00A61K9/107C12N15/87C08G83/00
Inventor 聂宇柯博文陈晓冰姜倩
Owner SICHUAN UNIV
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