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A kind of autofluorescence degradable polycitrate non-viral gene carrier and its preparation method

A polycitrate and autofluorescence technology, which can be applied to other methods of inserting foreign genetic materials, recombinant DNA technology, etc., can solve problems such as poor water solubility, and achieve increased stability, simple preparation method, and reduced toxicity and immunogenicity. Effect

Active Publication Date: 2020-03-17
XI AN JIAOTONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

POC is formed by the condensation reaction of citric acid and 1,8-octanediol. It has the advantages of non-toxic synthetic monomers, good biocompatibility, easy reaction conditions and no impurities introduced during the reaction process, easy post-molding, abundant supply, The price is cheap, and the monomeric citric acid of POC is also a metabolite of tricarboxylic acid in the human body, and the three carboxylic acid groups on it can be modified many times, but the surface of POC has no positive charge, and its water solubility is very poor, making it a gene The application of the carrier in the body is greatly limited. If the surface charge and water solubility of the POC can be improved by further modifying the POC, the application in the body will be greatly improved.

Method used

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  • A kind of autofluorescence degradable polycitrate non-viral gene carrier and its preparation method
  • A kind of autofluorescence degradable polycitrate non-viral gene carrier and its preparation method
  • A kind of autofluorescence degradable polycitrate non-viral gene carrier and its preparation method

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Embodiment 1

[0036] 1) Preparation of POCG prepolymer: Add 6 g of citric acid and 1,8-octanediol in a molar ratio of 1:1 into a 50 mL round-bottomed flask, stir in a nitrogen atmosphere and put it in an oil bath at 160 °C Melting; after the reaction monomers citric acid and 1,8-octanediol are completely melted, the temperature is immediately lowered to 140°C, and the reaction is carried out under nitrogen atmosphere for 5 hours. The reaction product was dialyzed and purified in deionized water for 2 days, and freeze-dried for later use;

[0037] 2) Preparation of POCG-PEI 600 polymer: weighed 0.5mmol polycitrate (POCG) was stirred in DMSO to dissolve it completely, then added 1.5mmol EDC, stirred at room temperature for 30min, then added 1.5mmol NHS, stirred at room temperature 12h, then add polyethyleneimine with a molecular weight of 600Da, stir at room temperature for 12h, and dialyze and purify the product in deionized water for 2 days to remove unreacted monomers, DMSO solvent, and ca...

Embodiment 2

[0040] 1) Preparation of POCG prepolymer: Add 6 g of citric acid, 1,8-octanediol and polyethylene glycol in a molar ratio of 1:0.4:0.6 into a 50 mL round-bottomed flask, stir and put into Melt in an oil bath at 160°C; after the reaction monomers citric acid, 1,8-octanediol and polyethylene glycol are all melted, immediately lower the temperature to 140°C, and react for 5 hours under a nitrogen atmosphere. The reaction product was dialyzed and purified in deionized water for 2 days, and freeze-dried for later use;

[0041] 2) Preparation of POCG-PEI 600 polymer: Weigh 0.5mmol water-soluble polycitrate (POCG) in 15mL, 50mM, pH 4-6 MES buffer and stir to dissolve completely, then add 1.5mmol EDC , stirred at room temperature for 30min, then added 1.5mmol NHS, stirred at room temperature for 12h, then added polyethyleneimine with a molecular weight of 600Da, stirred at room temperature for 12h, and purified the product by dialysis in deionized water for 2 days to remove unreacted ...

Embodiment 3

[0044] 1) Preparation of POCG prepolymer: Add 6 g of citric acid, 1,8-octanediol and polyethylene glycol in a molar ratio of 1:0.7:0.3 into a 50 mL round-bottomed flask, stir and put into Melt in an oil bath at 160°C; after the reaction monomers citric acid, 1,8-octanediol and polyethylene glycol are all melted, immediately lower the temperature to 140°C, and react for 5 hours under a nitrogen atmosphere. The reaction product was dialyzed and purified in deionized water for 2 days, and freeze-dried for later use;

[0045] 2) Preparation of POCG-PEI 600 polymer: Weigh 0.5mmol water-soluble polycitrate (POCG) in 15mL, 50mM, pH 4-6 MES buffer and stir to dissolve completely, then add 1.5mmol EDC , stirred at room temperature for 30min, then added 1.5mmol NHS, stirred at room temperature for 12h, then added polyethyleneimine with a molecular weight of 600Da, stirred at room temperature for 12h, and purified the product by dialysis in deionized water for 2 days to remove unreacted ...

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Abstract

The invention discloses a non-viral gene carrier of autofluorescence degradable poly-citrate and a preparation method thereof. Citric acid, 1,8-octylene glycol and polyethylene glycol are subjected to thermal polymerization to obtain a POCG prepolymer; then, the POCG and PEI (polyethyleneimine) are subjected to catalytic polymerization to obtain a POCG-PEI polymer; the POCG-PEI polymer and various genes (DNA / siRNA / miRNA) can form a stable nanocomposite in HEPES buffer liquid. The used thermal polymerization method has the advantages that the environment is protected; the operation is convenient; the raw material cost is low. The prepared POCG-PEI has good biocompatibility and high gene transfection efficiency; meanwhile, the POCG-PEI also shows certain fluorescence characteristics and optical stability, so that the polymer has good application prospects in the gene treatment process.

Description

【Technical field】 [0001] The invention belongs to the technical field of degradable biomedical materials, and in particular relates to a non-viral gene carrier of autofluorescent degradable polycitrate and a preparation method thereof. 【Background technique】 [0002] In recent years, with the continuous deepening of gene therapy research, the key to the success of gene therapy is to develop vectors with targeted, controllable and effective expression of the target gene. Although about 80% of the vectors currently used in gene therapy are viral vectors, there are many safety issues in their use due to their high immunogenicity and potential carcinogenicity, but non-viral vectors can be well To solve these problems, and the packaging size is small and the cost is low, so non-viral vectors are attracting more and more attention. [0003] At present, for the use of synthetic biodegradable polymer materials, polyester biomaterials have become the most promising type of biomateri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/87C08G81/00
CPCC08G81/00C12N15/87
Inventor 雷波王敏郭旖
Owner XI AN JIAOTONG UNIV
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