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A kind of modified hyperbranched polyethyleneimine, its preparation method and application

A polyethyleneimine, modified technology, applied in other methods of inserting foreign genetic materials, pharmaceutical formulations, genetic material components, etc., can solve problems such as troublesome preparation, not much improvement in transfection efficiency, and reduce the charge density. , Excellent biocompatibility, overcoming high toxicity

Active Publication Date: 2019-03-12
长春金传科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These modifications reduce cytotoxicity and transfection efficiency to a certain extent, but the preparation of such polymers is relatively troublesome, and the transfection efficiency is not greatly improved

Method used

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  • A kind of modified hyperbranched polyethyleneimine, its preparation method and application
  • A kind of modified hyperbranched polyethyleneimine, its preparation method and application
  • A kind of modified hyperbranched polyethyleneimine, its preparation method and application

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preparation example Construction

[0040] The present invention also provides a kind of preparation method of above-mentioned modified polyethyleneimine, comprising:

[0041] When-R 1 for -OH or R 3 -NH-, and R 3 When the amino group is not included, the hyperbranched polyethylenimine and the compound shown in formula (I) are dissolved in an organic solvent, and react under the conditions of the presence of a condensing agent and an activator and anhydrous and oxygen-free conditions to obtain a modified hyperbranched polyethylenimine. Ethylene imine.

[0042] Wherein, the hyperbranched polyethyleneimine and the compound represented by formula (I) are the same as described above, and will not be repeated here.

[0043] Dissolving the compound shown in hyperbranched polyethyleneimine and formula (I) in an organic solvent, wherein the organic solvent is an organic solvent well known to those skilled in the art, without special restrictions, preferably in the present invention DMF (N,N-Dimethylformamide).

[0...

Embodiment 1

[0088] Dissolve hyperbranched polyethyleneimine, p-toluenesulfonyl (Tos) and tert-butoxycarbonyl (Boc) double-protected arginine ((Boc)Arg(Tos)-OH) in DMF, In the presence of agents EDC and NHS (the moles of the condensing agent and the activator are 1.5 times the moles of (Boc)Arg(Tos)-OH), react at 25°C for 24h under anhydrous and oxygen-free conditions to obtain hyperbranched polyethylene Amine grafted (Boc)Arg(Tos) polymer (PEI-(Boc)Arg(Tos)). Then remove the Boc group with trifluoroacetic acid to obtain the final product PEI-Arg (Tos), which is modified hyperbranched polyethyleneimine.

[0089] Utilize nuclear magnetic resonance to analyze the modified hyperbranched polyethyleneimine obtained in embodiment 1, obtain its nuclear magnetic resonance spectrum (hydrogen spectrum), as figure 1 shown.

[0090] The grafting ratio of p-toluenesulfonyl-protected arginine (Arg(Tos)) to hyperbranched polyethyleneimine (PEI) obtained in Example 1 is listed in Table 1.

[0091] Corr...

Embodiment 2

[0094] In Vitro Transfection of HeLa Cells Mediated by PEI-Arg(Tos) pGL3 (Luciferase Plasmid), pEGFPN1 (Green Fluorescent Protein Plasmid) and pCMV-lac Z (β-Galactosidase Plasmid)

[0095] 1) Culture of HeLa cells

[0096] Human cervical cancer cells (HeLa cells) were cultured continuously in a medium containing 10% calf serum (5% CO 2 , 37°C incubator).

[0097] 2) In vitro transfection

[0098] Within 24 hours before transfection, cells in the logarithmic growth phase were collected, digested with trypsin and diluted with Dulbecco’s modified Eagle’s medium at 1×10 per well. 4 Cells were seeded in 96-well culture plates at a density of 37°C in an incubator containing 5% carbon dioxide by volume and continued to culture until the confluence reached 80%-90%. During transfection, aspirate and discard the culture medium in the cell culture plate added the day before, wash twice with phosphate buffered saline, the aqueous solution of complex particles (PEI-Arg(Tos) and plasmid)...

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Abstract

The present invention provides a kind of modified hyperbranched polyethyleneimine, its preparation method and application, and this modified hyperbranched polyethyleneimine is obtained by the compound represented by hyperbranched polyethyleneimine graft formula (I); wherein , the -R1 is -OH or R3-NH-; the -R3 is a group formed by the amino acid losing its terminal amino group; the -R2 is methyl or p-tolyl. Compared with the prior art, the present invention introduces methanesulfonyl or p-toluenesulfonyl to hyperbranched polyethyleneimine, which can effectively reduce the charge density without reducing the overall charge, so that the modified hyperbranched poly Ethyleneimine has a branched structure and concentrated charge density, which overcomes the shortcomings of high toxicity of polyethyleneimine homopolymer and insufficient concentration of linear polymer-grafted polyethyleneimine; while modifying hyperbranched polyethyleneimine It has a strong ability to load genetic material and can freely adjust the charge density. It is an efficient gene delivery carrier with excellent biocompatibility.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a modified hyperbranched polyethyleneimine, a preparation method and an application. Background technique [0002] With the continuous advancement of bioscience and technology, gene therapy will play an important role in the process of overcoming chronic diseases such as cancer and genetic diseases, and will gradually become a common and effective method (see Jeong JH, Kim SW, ParkTG.Park. Molecular design of functional polymers for genetherapy. Prog. Polym. Sci. 2007; 32(11):1239-1274.). [0003] Gene therapy refers to a new biomedical technology that introduces human normal genes or genes with therapeutic effects into human target cells in a certain way to correct gene defects or exert therapeutic effects, so as to achieve the purpose of treating diseases. Successful gene therapy depends on effective gene carriers, and common carriers are divided into viral vectors and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G73/02C12N15/87A61K48/00
CPCA61K48/0041C08G73/0206C08G73/0213C12N15/87
Inventor 田华雨郭兆培陈学思林琳陈杰孙平杰
Owner 长春金传科技有限公司
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