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Preparation method for N-2-hydroxypropyldimethylethyl ammonium chloride chitosan/carboxymethyl chitosan Newcastle disease virus nanoparticles

A technology of hydroxypropyl dimethyl ethyl ammonium chloride and epoxypropyl dimethyl diethyl ammonium chloride, applied in N-2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan /In the field of preparation of carboxymethyl chitosan Newcastle disease virus nanoparticles, it can solve the problems of poor adsorption capacity on mucosal surface, low vaccine antigen encapsulation rate and drug loading capacity, poor solubility of chitosan, etc.

Active Publication Date: 2015-07-22
HEILONGJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The present invention aims to solve the problems of poor solubility and low degree of substitution of chitosan, resulting in low encapsulation efficiency and drug loading capacity of chitosan nanoparticles to vaccine antigens, and poor adsorption capacity on the surface of mucous membranes, and provides a kind of N- Preparation method of 2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan / carboxymethyl chitosan Newcastle disease virus nanoparticles

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  • Preparation method for N-2-hydroxypropyldimethylethyl ammonium chloride chitosan/carboxymethyl chitosan Newcastle disease virus nanoparticles
  • Preparation method for N-2-hydroxypropyldimethylethyl ammonium chloride chitosan/carboxymethyl chitosan Newcastle disease virus nanoparticles
  • Preparation method for N-2-hydroxypropyldimethylethyl ammonium chloride chitosan/carboxymethyl chitosan Newcastle disease virus nanoparticles

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specific Embodiment approach 1

[0024] Specific embodiment one: the preparation method of present embodiment N-2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan / carboxymethyl chitosan Newcastle disease virus nanoparticle, carry out according to the following steps:

[0025] One, the preparation of N-2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan (N-2-HFCC);

[0026] Two, the preparation of N-2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan / carboxymethyl chitosan Newcastle disease virus nanoparticles (NDV / LaSota-N-2-HFCC / CMC-NPs):

[0027] a. Take 0.5 ~ 2.0mg / mL N-2-hydroxypropyldimethylethylammonium chloride chitosan solution 5mL, drop 2mL of Newcastle disease L-series virus solution into it, stir magnetically for 3min and mix evenly to obtain a solution A;

[0028] b. Stir solution A magnetically at 900-1400 r / min for 10-50 min at room temperature under sterile conditions, then add 0.5-3.0 mg / mL N,O-carboxymethyl chitosan solution 1.25-7.5 mL, to obtain solution B;

[0029] c....

specific Embodiment approach 2

[0032] Specific embodiment two: the present embodiment is different from specific embodiment one: the preparation method of N-2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan in the step one is as follows:

[0033] One, the synthesis of epoxypropyl dimethyl ethyl ammonium chloride (EEA):

[0034]Add epichlorohydrin to a dry three-neck flask equipped with an electric stirrer, dropping funnel and condensing reflux device at 4-15°C, stir continuously and add N,N-dimethylethylamine dropwise, and finish adding within 1 hour, at the same time Heat to 10-35°C, react at constant temperature for 4-14 hours, stop the reaction, wash the product with ice acetone, and filter it with suction, put the filtered product in a watch glass, and dry it in a vacuum dryer for 6-24 hours to obtain ring Oxypropyldimethylethylammonium chloride; wherein the ratio of epichlorohydrin to N,N-dimethylethylamine is 2 to 6:1;

[0035] Two, the preparation of N-2-hydroxypropyl dimethyl ethyl ammonium...

specific Embodiment approach 3

[0044] Specific embodiment three: this embodiment is different from specific embodiment one or two: the virus content of the Newcastle disease L-line virus liquid in step 2a is 10 7.5 EID 50 . Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a preparation method for N-2-hydroxypropyldimethylethyl ammonium chloride chitosan (N-2-HFCC) / carboxymethyl chitosan Newcastle disease virus nanoparticles, relates to preparation methods for virus nanoparticles, and solves problems that chitosan nanoparticles have low encapsulation rate and drug-loading capacity on vaccine antigens and poor adsorption capability on the mucosal surface. The method comprises the following steps: 1, preparation of N-2-HFCC; 2, preparation of the N-2-HFCC / carboxymethyl chitosan Newcastle disease virus nanoparticles, wherein a, a Newcastle disease L-series virus solution is dropwise added into an N-2-HFCC solution, stirring is performed, and a solution A is obtained; b, the solution A is stirred, an N,O-carboxymethyl chitosan solution is dropwise added, and a solution B is obtained; c, the solution B is centrifuged, precipitated, washed with water and dried to obtain the N-2-HFCC / N,O-carboxymethyl chitosan nanoparticles loaded with Newcastle disease attenuated live vaccines. The nanoparticles prepared with the method are high in biosafety, good in bacteriostatic effect and high in encapsulation rate. The method is used for preparing the Newcastle disease virus nanoparticles.

Description

technical field [0001] The invention relates to a preparation method of N-2-hydroxypropyldimethylethylammonium chloride chitosan / N, O-carboxymethyl chitosan nanoparticles loaded with attenuated live Newcastle disease vaccine. Background technique [0002] Newcastle Disease (ND) is a highly contagious infectious disease caused by Newcastle Disease Virus (NDV). Be the first line of defense against NDV infection. Therefore, improving the defense ability of the mucosal immune system can effectively prevent the adsorption and invasion of NDV. At present, the commonly used vaccines for ND vaccination are traditional attenuated live vaccines and inactivated vaccines, but both of them have certain limitations in practical application. How to improve the immune effect of vaccines is an urgent subject to be overcome in vaccine immunization research. , The transformation of vaccine formulations and delivery methods is one of the methods to improve the immune effect of vaccines. Curr...

Claims

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Application Information

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IPC IPC(8): A61K39/17A61K47/36A61P31/14C08B37/08
Inventor 赵凯王晓华
Owner HEILONGJIANG UNIV
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