Hyaluronic acid (HA)-polyethyleneimine (PEI) bonded copolymer as well as preparation method and application thereof

A technology of polyethyleneimine and hyaluronic acid, which is applied in the field of biomedical materials, can solve problems such as limited application, and achieve the effects of prolonging the action time, improving safety, and enhancing adhesion

Inactive Publication Date: 2012-04-11
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Furthermore, the inherent cytotoxicity of PEI limits its application in in vivo gene therapy [16]

Method used

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  • Hyaluronic acid (HA)-polyethyleneimine (PEI) bonded copolymer as well as preparation method and application thereof
  • Hyaluronic acid (HA)-polyethyleneimine (PEI) bonded copolymer as well as preparation method and application thereof
  • Hyaluronic acid (HA)-polyethyleneimine (PEI) bonded copolymer as well as preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1b

[0053] The synthesis of embodiment 1bPEI-HA complex

[0054] a. Dissolve 10 mL of HA with a concentration of 2 mg / mL in 10 mL of bPEI with a concentration of 40 mg / mL, and adjust the pH value of the reactant to 6.5 with 1.0 mol / L HCl solution.

[0055] b. Dissolve 40.4 mg EDC·HCl and 28.5 mg HOBt in 1 ml of 50% by volume DMSO solution.

[0056] c. Add the solution obtained in step b to the solution obtained in step a. After stirring and mixing at room temperature for 24 hours, adjust the pH to 7.0 with NaOH with a concentration of 1.0 mol / L.

[0057] d. Pour the solution obtained in step c into a cleaned 10kDa dialysis bag, dialyze in 100mmol / L NaCl solution for 2 days, dialyze in 25wt% ethanol for one day, and dialyze in purified water for one day to obtain 1 μg / μL of bPEI-HA complex The solution was freeze-dried to obtain a white powdery solid. reuse 1 H-NMR analysis detected the degree of bPEI modification of HA.

[0058] use 1 H-NMR analysis to detect the degree of bP...

Embodiment 2b

[0059] The effect detection experiment of embodiment 2bPEI-HA complex

[0060] (1) Preparation of siRNA / bPEI-HA complex

[0061] Take the anti-PGL3-Luc siRNA with a concentration of 0.75 μg / μL and the bPEI-HA complex with a concentration of 0.75 μg / μL and mix them in volume ratios of 1:1, 1:3 and 1:5, respectively. As shown in Table 1, incubate at room temperature for 15 minutes to make siRNA / bPEI-HA complex. Finally, 3 μL of 5 mol / L NaCl solution was added to make the final concentration of siRNA / bPEI-HA complex reach 150 mmol / L.

[0062] Mix 0.75 μg / μL anti-PGL3-Luc siRNA with 0.75 μg / μL bPEI at a volume ratio of 1:1, 1:3, and 1:5 respectively. The mixing method is shown in Table 2, and incubated at room temperature 15 minutes to make siRNA / bPEI complexes. Finally, 3 μL of 5 mol / L NaCl solution was added to make the final concentration of the siRNA / bPEI complex reach 150 mmol / L.

[0063] Table 1. Mixing ratio of siRNA and bPEI-HA complex

[0064]

[0065] Table 2. Mi...

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Abstract

The invention relates to a hyaluronic acid (HA)-polyethyleneimine (PEI) bonded polymeric carrier material as well as a preparation method and application thereof. The polymeric carrier material is an HA-PEI polymeric carrier material prepared through grafting HA used as a frame onto PEI by utilizing 1-Ethyl-3-(3-dimethyllaminopropyl)carbodiimide hydrochloride (EDC.HCl) as a cross-linking agent. The polymeric carrier material can be used for improving the targetting performance of gene/anion proteins in a cell, reducing the toxicity to the cell and improving the conversion efficiency of targetting genes, and has a wide application prospect.

Description

technical field [0001] The invention relates to a polymeric carrier material, which belongs to the field of biomedical materials, in particular to a hyaluronic acid-bonded polyethylenimine copolymer, its preparation method and application. Background technique [0002] The discovery and research of the RNA interference (RNA interference, RNAi) phenomenon has brought new opportunities for gene therapy, and it is rapidly developing into the main method of gene therapy. Double-stranded small interference RNA (small interference RNA, siRNA) consisting of 21-23 base pairs [1，2] , can specifically degrade the messenger RNA (mRNA) complementary to its base pair in the cytoplasm of animals, thereby blocking the expression of mRNA-encoded protein and realizing gene silencing [3，4] , that is, RNA interference. However, siRNA will be rapidly digested in the body, and the uptake of siRNA by cells is very small, so the gene silencing efficiency of siRNA in vivo is very low [5-7] . In...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08C08G73/04A61K47/48A61K47/36A61K48/00A61P9/00A61P11/06A61P35/00C12N15/87
Inventor 蒋革冯晓娜董爽王慧国
Owner DALIAN UNIV
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