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Hyaluronic acid nanometer vesicle, and preparation method and application thereof

A technology of hyaluronic acid and nanovesicles, applied in the field of biomedicine, can solve the problems of unfavorable therapeutic gene delivery, no public report of hyaluronic acid nanovesicles, short half-life, etc., and achieves regular morphology, low toxicity and cost. low cost effect

Active Publication Date: 2017-05-31
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cationized gene delivery vectors also have their insurmountable disadvantages: due to the effect of positive charge, they quickly combine with plasma proteins and are quickly cleared by the reticuloendothelial system in the body, so the half-life in the body is very short, which is not conducive to treatment. Gene delivery (Cell Research, 2015, 25:237)
However, there has been no public report on hyaluronic acid nanovesicles and its use as a gene delivery carrier.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] The present invention can be made by following method in concrete implementation:

[0016] Add 150mg of hyaluronic acid to 15mL of N,N-dimethylformamide, add 138mg of activating reagent ethyl-(3-dimethylpropyl)carbodiimide hydrochloride and 84mg of N-hydroxybutanedi Imide, protected from light, stirred and reacted at room temperature for 4 hours under nitrogen protection, then added 250 mg of long-chain diamine, stirred and reacted at room temperature for 48 hours, added 95% ethanol for dialysis for 24 hours, then dialyzed with ultrapure water for 48 hours, and freeze-dried to obtain Functional long-chain modified hyaluronic acid; dissolve the above product in 15 mL of ultrapure water, add dropwise 1ml of hydrochloric acid with a mass concentration of 3.7%, and then add 500 mg of polyethyleneimine to react for 48 hours, and dialyze the product with ultrapure water for 3 days. Freeze-dry to obtain hyaluronic acid nanovesicles.

Embodiment 2

[0018] The present invention can be made by following method in concrete implementation:

[0019] Add 120 mg of hyaluronic acid to 12 mL of N,N-dimethylformamide, add 30 mg of activating reagent ethyl-(3-dimethylpropyl) carbodiimide hydrochloride and 20 mg of maleic anhydride, Protected from light, stirred at room temperature for 3 hours under nitrogen protection, then added 220 mg of amino-terminal long-chain alcohol, stirred at room temperature for 40 hours, added 95% ethanol for dialysis for 24 hours, then dialyzed with ultrapure water for 48 hours, and freeze-dried to obtain functional long-chain Modified hyaluronic acid: Dissolve the above product in 15 mL of ultrapure water, add 30 mg of activating reagent ethyl-(3-dimethylpropyl) carbodiimide hydrochloride and 20 mg maleic anhydride to activate for 4 h , adding 400 mg of polyethyleneimine to react for 36 hours, the product was dialyzed with ultrapure water for 3 days, and freeze-dried to obtain hyaluronic acid nanovesic...

Embodiment 3

[0021] The present invention can be made by following method in concrete implementation:

[0022] Add 170mg of hyaluronic acid to 17mL of N,N-dimethylformamide, add 150mg of activating reagent ethyl-(3-dimethylpropyl)carbodiimide hydrochloride and 100mg of N-hydroxybutyl Diimide, protected from light, stirred at room temperature for 5 hours under nitrogen protection, then added 270 mg of amino-terminal long-chain acid, stirred at room temperature for 55 hours, added 95% ethanol for dialysis for 24 hours, then dialyzed with ultrapure water for 48 hours, and freeze-dried , to obtain functional long-chain modified hyaluronic acid; the above product was dissolved in 15 mL of ultrapure water and activating reagent 150 mg of ethyl-(3-dimethylpropyl) carbodiimide hydrochloride and 100 mg of N After activating -hydroxysuccinimide for 4 hours, react with 600 mg of polyethyleneimine for 72 hours, dialyze the product with ultrapure water for 3 days, and freeze-dry to obtain hyaluronic ac...

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Abstract

The invention relates to a hyaluronic acid nanometer vesicle, and a preparation method and application thereof, which can effectively solve the preparation problem of the hyaluronic acid nanometer vesicle, and realizes the application in tumor gene therapy drugs. The hyaluronic acid nanometer vesicle is characterized in that hyaluronic acid is a functional long chain grafted through a chemical bond, polyethyleneimine is grafted to the tail end of the functional long chain, when being interacted with nucleic acid, the polyethyleneimine compresses the nucleic acid through electrostatic interaction, and the hyaluronic acid fully wraps the periphery of the polyethyleneimine / nucleic acid to form a vesicle structure. The raw material source is wide, the preparation method is simple, the cost is low, and the prepared hyaluronic acid nanometer vesicle is regular in morphology, has the particle size range ranging from 100 to 300nm, is stable in structure, uniform in distribution, good in biocompatibility and low in virulence, and can load a macromolecule mass nucleic acid antitumor drug, realize highly uptake and transfect to a tumor cell so as to realize the application in the tumor therapy drugs.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a hyaluronic acid nanovesicle and its preparation method and application. Background technique [0002] Studies have proved that many human diseases are closely related to changes in gene function or structure, and gene therapy has become an emerging medical treatment to improve human health. The key to gene therapy is to select the appropriate gene carrier, so that the therapeutic gene can be delivered to the target cells efficiently and quickly, and realize its biological function. Gene carriers are mainly divided into viral gene carriers and non-viral gene carriers (Current Drug Delivery, 2004, 1:165). Compared with viral gene vectors, non-viral gene vectors have the advantages of low toxicity, no immunity, easy preparation, and suitable for in vivo research, and are widely used. [0003] Due to the presence of phosphate groups, nucleic acid molecules such as DNA and RNA are usual...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/127A61K48/00A61K31/7088A61K47/36A61K47/34A61P35/00
CPCA61K9/1273A61K31/7088A61K47/34A61K47/36A61K48/00
Inventor 任雪玲张振中林静张红岭武园园刘晓张瑞孟二娟韩淼
Owner ZHENGZHOU UNIV