Preparation method and application of enzyme-supported chitosan nanoparticle

A chitosan nanometer and nanoparticle technology is applied in the directions of non-active ingredients medical preparations, medical preparations containing active ingredients, pharmaceutical formulas, etc. Low-level problems, to achieve the effect of non-toxic components, small damage to protein molecules, and avoiding degradation

Inactive Publication Date: 2015-04-29
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, horseradish peroxidase (HRP) has the problems of low delivery efficiency, p

Method used

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  • Preparation method and application of enzyme-supported chitosan nanoparticle
  • Preparation method and application of enzyme-supported chitosan nanoparticle
  • Preparation method and application of enzyme-supported chitosan nanoparticle

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Embodiment 1. Preparation of enzyme-loaded chitosan nanoparticles (HRP-CS-NPs)

[0066] First, 0.5 mg of horseradish peroxidase (HRP) was dissolved in 1 mL of ultrapure water, and 50 mg of the chemical cross-linker 1-ethyl-3-[3-dimethylaminopropyl]-carbonated Diimine (EDC) was magnetically stirred in the above mixture for 30 minutes to activate the carboxyl groups on the surface of horseradish peroxidase (HRP). Then, 10 mg of chitosan (Chitosan) was dissolved in 1 mL of 1% acetic acid solution, and the pH was adjusted to 5.0 after magnetic stirring at 60° C. for complete dissolution. Finally, mix the 10mg / mL chitosan solution with the above-mentioned activated horseradish peroxidase (HRP) mixture, add 40mg of N-hydroxysuccinimide (NHS), and stir magnetically at 4°C for 48h To obtain the coupling complex (HRP-CS) of chitosan molecules and horseradish peroxidase (HRP).

[0067] Add 5 mL of chitosan molecule and horseradish peroxidase (HRP) coupling complex (HRP-CS) with...

Embodiment 2

[0069] Embodiment 2. The influence of different acetic acid concentrations on the preparation of chitosan nanoparticles

[0070] The chitosan of 100mg is dissolved in the acetic acid solution of 100mL (0.5%, 0.8%, 1%, 1.2%, 1.5%, 2%) respectively, is mixed with the chitosan solution of 1mg / mL, and magnetic force stirs to dissolve overnight, And regulate chitosan solution pH to 5.0 with 20wt% sodium hydroxide, and remove the insoluble impurity in the solution with the microporous membrane of 0.22 μm, make chitosan stock solution; Prepare the sodium tripolyphosphate (TPP) of 1mg / mL Aqueous solution, through a 0.22 μm microporous membrane to remove insoluble impurities in the solution to prepare TPP stock solution.

[0071] Chitosan nanoparticles (CS-NPs) preparation: 5 mL of chitosan (CS) with a pH of 5.0 at 1 mg / mL was added to a 10 mL beaker and stirred, the stirring speed was controlled to be 500 rpm, and the stirring ambient temperature was about 10 ° C. Add 1 mL of 1 mg / mL...

Embodiment 3

[0073] Embodiment 3. The impact of different chitosan concentrations on the preparation of chitosan nanoparticles

[0074] Take a certain amount of chitosan nanoparticles and dissolve them in 1% acetic acid solution, and prepare the concentrations as (0.5mg / mL, 0.8mg / mL, 1mg / mL, 1.5mg / mL, 1.8mg / mL, 2mg / mL) of the chitosan solution, magnetically stirred overnight and dissolved, and adjusted the pH of the chitosan solution to 5.0 with 20wt% sodium hydroxide, and removed the insoluble impurities in the solution with a microporous membrane of 0.22 μm to prepare the chitosan stock solution; prepare a 1 mg / mL sodium tripolyphosphate (TPP) aqueous solution, and remove insoluble impurities in the solution through a 0.22 μm microporous membrane to obtain a TPP stock solution.

[0075] Chitosan nanoparticles (CS-NPs) preparation: the concentration of (0.5mg / mL, 0.8mg / mL, 1mg / mL, 1.5mg / mL, 1.8mg / mL, 2mg / mL) shell at pH 5.0 Add 5mL of polysaccharide (CS) solution into a 10mL beaker and ...

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Abstract

The invention discloses a preparation method of an enzyme-supported chitosan nanoparticle. The preparation method of the enzyme-supported chitosan nanoparticle comprises the following steps: (1) preparing a crosslinking complex of chitosan and enzyme; and (2) preparing nanoparticles by adopting an ionic gel method, stirring the crosslinking complex obtained in the step (1) by virtue of a magnetic stirrer, then dropwise adding sodium tripolyphosphate, obtaining a uniform opalescent suspension after reaction is finished, and then carrying out centrifugal washing for multiple times, so that pure enzyme-supported chitosan nanoparticles are obtained. The preparation method of the enzyme-supported chitosan nanoparticle has the advantages that firstly the enzyme-supported chitosan nanoparticle is good in stability, good in biocompatibility, high in drug loading rate and high in enzyme wrapping efficiency when used in an enzyme-precrusor mediation process; secondly, the preparation process is simple, reaction conditions are mild, convenience and rapidness are realized, ingredients are nontoxic, the drug encapsulation efficiency is high, and damage on protein molecules is small; and thirdly, chitosan nanoparticles wrap HRP (horse reddish peroxidase) inside, degradation of HRP in vivo is avoided, and a certain degree of controlled release is achieved.

Description

technical field [0001] The invention relates to the technology for preparing composite biomedical materials in the field of polymer material science, and more specifically relates to the preparation and application of enzyme-loaded chitosan nanoparticles. Background technique [0002] In recent decades, chemotherapy has made significant progress in the treatment of tumors, and has become a basic means of treatment for malignant tumors, thus significantly prolonging the survival time of patients. However, some existing chemotherapeutic drugs, in the process of administration, will greatly damage the normal cells of the human body while killing tumor cells, so that there will be obvious toxic and side effects in the treatment, which will affect the curative effect of the drugs, and bring harm to the patients. to great pain. Therefore, drug carriers have obvious advantages in tumor treatment, especially nano drug delivery system has become a research hotspot at home and abroad...

Claims

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Application Information

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IPC IPC(8): A61K9/14A61K47/48A61K38/44A61K38/46A61K38/47A61P35/00A61K31/405A61K47/61
Inventor 王平陈超曹晓丹余海军
Owner EAST CHINA UNIV OF SCI & TECH
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