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Polyethylene glycol monomethyl ether-poly 2-methyl-carboxyl propylene carbonate graft polyethyleneimine copolymer, preparation method thereof and application thereof

A technology of polyethyleneimine copolymer and polyethylene glycol monomethyl ether, which is applied to polyethyleneimine copolymer and polyethylene glycol monomethyl ether-poly2-methyl-2-carboxypropylene carbonate grafted with polyethyleneimine. The preparation method and application field can solve the problems of non-degradability and high cytotoxicity, and achieve the effects of high use value, high transfection efficiency and low cytotoxicity.

Inactive Publication Date: 2010-02-03
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the pure use of homopolymer PEI as a gene carrier is often limited by its non-degradability and high cytotoxicity.

Method used

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  • Polyethylene glycol monomethyl ether-poly 2-methyl-carboxyl propylene carbonate graft polyethyleneimine copolymer, preparation method thereof and application thereof
  • Polyethylene glycol monomethyl ether-poly 2-methyl-carboxyl propylene carbonate graft polyethyleneimine copolymer, preparation method thereof and application thereof
  • Polyethylene glycol monomethyl ether-poly 2-methyl-carboxyl propylene carbonate graft polyethyleneimine copolymer, preparation method thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Preparation of polyethylene glycol monomethyl ether-poly 2-methyl-2-carboxypropylene carbonate block copolymer

[0038] Respectively according to the charging amount listed in Table 1, take by weighing polyethylene glycol monomethyl ether (mPEG) with a number average molecular weight of 5000 5000 ) and 2-methyl-2-benzyloxycarbonyl propylene carbonate (MBC), the two are fed into different dry reactors, respectively add 300ml of anhydrous toluene at 130 ° C to azeotropically remove water for 5 hours, azeotropically After the water removal is completed, part of the toluene is evaporated at a temperature of 130°C, and the volume of the remaining toluene is 50ml as the end point of each distillation; the catalyst diethylzinc is dissolved in anhydrous toluene at a molar concentration of 0.2mmol / ml, respectively Add diethylzinc / toluene solution to the corresponding reactor according to the diethylzinc charging amount in table 1, and follow the polymerization temp...

Embodiment 2

[0041] Embodiment 2: Preparation of polyethylene glycol monomethyl ether-poly2-methyl-2-carboxypropylene carbonate grafted polyethyleneimine copolymer

[0042] Weigh the corresponding feeding amount according to the type of mPEG-b-PMCC listed in Table 2, feed into different reactors together with the dimethyl sulfoxide of the corresponding reaction volume described in Table 2, stir and dissolve, According to the type of polyethyleneimine (PEI) shown in Table 2 and the molar amount of feed, PEI was weighed respectively and added to the corresponding reactor, and then according to the molar amount of feed shown in Table 2, 1-(3 -Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC.HCl) is also added to the corresponding reactor, the temperature is controlled at 25°C, and the reaction is carried out with stirring according to the reaction time in Table 2. , after the reaction was completed, the reaction solution was respectively loaded into a dialysis bag with a molecular ...

Embodiment 3

[0045] Example 3: Polyethylene glycol monomethyl ether-poly 2-methyl-2-carboxypropylene carbonate grafted polyethyleneimine copolymer mediated luciferase plasmid to African green monkey kidney cells (Cos-7 cells) in vitro transfection

[0046] 1. Cos-7 cell culture

[0047] The Cos-7 cells were placed in a culture solution containing 10% fetal bovine serum by volume, and continuously cultured in an incubator containing 5% carbon dioxide by volume at 37°C.

[0048] 2. In vitro transfection

[0049] (1) No serum transfection

[0050] Within 24 hours before transfection, take Cos-7 cells in the logarithmic growth phase, digest with trypsin and dilute with DMEM 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, discard the culture medium in the cell culture plate added the day before, wash twice with phosphat...

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Abstract

The invention relates to polyethylene glycol monomethyl ether-poly 2-methyl-carboxyl propylene carbonate graft polyethyleneimine copolymer, a preparation method thereof and application thereof. The preparation method comprises the step of directly condensing carboxyl in the polyethylene glycol monomethyl ether-poly 2-methyl-carboxyl propylene carbonate graft polyethyleneimine segmented copolymer with amino in polyethyleneimine to form the graft copolymer. The copolymer is a polycation gene carrier, integrates the advantages of polyethylene glycol, Makrolan and polyethyleneimine, and has high transfection efficiency, wherein the highest transfection efficiency to the medium luciferase plasmid of african green monkey kidney cell is 14 times of that of American Invitrogen biological company transfection reagent Lipofetamine<TM>2000, can effectively antagonize the inhibiting effect of blood serum to the transfection, and has less cell toxicity, wherein consistency thereof is not higher than 200 microgramme / ml and cell survival rate is more than 80%.

Description

technical field [0001] The invention relates to polyethylene glycol monomethyl ether-poly2-methyl-2-carboxypropylene carbonate grafted polyethyleneimine copolymer, its preparation method and application. Background technique [0002] Gene therapy is expected by scientists to become an important medical method to solve difficult diseases in the future, but how to efficiently deliver therapeutic gene fragments into the lesion cells has been troubled. Viral vectors widely used in clinical practice currently have major safety hazards, which caused the first death of a patient in the history of gene therapy and the famous French "bubble baby" incident. Compared with the insecurity of viral gene carriers, artificially synthesized cationic polymers have gradually become a research hotspot in this field due to their non-immunogenicity, diverse molecular designs, controllable size and ability to connect targeting substances. [0003] Among the many polycationic carriers that have be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G81/00C08G64/18
Inventor 陈学思陈磊田华雨郭兆培陈杰林琳黄宇彬景遐斌
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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