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Preparation method of N-2-hydroxypropyl trimethyl ammonium chloride chitosan/N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle

A technology of hydroxypropyl trimethyl ammonium chloride and carboxymethyl chitosan, which is applied in the directions of antiviral agents, viral antigen components, powder transportation, etc., can solve the problems such as application limitations of chitosan, and achieve good biological adhesion. The effect of stability, uniform particle size distribution, and low drug toxicity and side effects

Active Publication Date: 2012-04-25
HEILONGJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] As the only natural polysaccharide with cations in nature, chitosan has special functions such as good biological activity, biocompatibility, degradability and antibacterial properties. However, chitosan can only be dissolved in acidic solution. Applications in this field are severely limited

Method used

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  • Preparation method of N-2-hydroxypropyl trimethyl ammonium chloride chitosan/N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle
  • Preparation method of N-2-hydroxypropyl trimethyl ammonium chloride chitosan/N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle
  • Preparation method of N-2-hydroxypropyl trimethyl ammonium chloride chitosan/N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle

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specific Embodiment approach 1

[0017] Specific embodiment 1: The preparation method of N-2-hydroxypropyltrimethylammonium chloride chitosan / N,O-carboxymethyl chitosan Newcastle disease live attenuated vaccine nanoparticles according to the following steps Realize:

[0018] 1. Take 5mL of N-2-hydroxypropyltrimethylammonium chloride chitosan solution with a concentration of 0.5~20mg / mL, and then add 1.25~7.5mL of Newcastle disease L-series virus solution dropwise to it, and magnetically stir for 3min , Obtain solution A;

[0019] 2. Solution A is magnetically stirred at 900~1300r / min for 10~40min under aseptic conditions at room temperature, and then 2mL N,O-carboxymethyl chitosan solution with a concentration of 0.5~3.0mg / mL is added dropwise to obtain the solution B;

[0020] 3. Centrifuge solution B at 4°C and 12000r / min for 2min, take the precipitate and wash it with deionized water three times, then freeze-dry it under vacuum for 24h to obtain N-2-hydroxypropyltrimethylammonium chloride chitosan / N, O-carboxy...

specific Embodiment approach 2

[0028] Specific embodiment two: this embodiment is different from specific embodiment one in that in step one, take 5mL of N-2-hydroxypropyltrimethylammonium chloride chitosan solution with a concentration of 1mg / mL, and then drop it into it 2mL Newcastle disease L-line virus solution. Other steps and parameters are the same as in the first embodiment.

specific Embodiment approach 3

[0029] Specific embodiment three: this embodiment is different from specific embodiment one in that in step two, solution A is magnetically stirred at 1200 r / min for 30 min under sterile conditions at room temperature, and then 2 mL of N, O- with a concentration of 1 mg / mL is added dropwise. Carboxymethyl chitosan solution. The other steps and parameters are the same as in the first embodiment.

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Abstract

The invention relates to a preparation method of a newcastle disease attenuated live vaccine nanoparticle, in particular to a preparation method of a N-2-hydroxypropyl trimethyl ammonium chloride chitosan / N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle, which comprises the following steps of: 1) dripping newcastle disease L-series virus fluid in N-2-hydroxypropyl trimethyl ammonium chloride chitosan solution to obtain solution A; 2) mixing the solution A, dripping N, O-carboxymethyl chitosan solution to obtain solution B; and 3) centrifuging the solution B, sedimenting, washing by deionized water, and carrying vacuum freezing and drying to obtain the product. According to the N-2-hydroxypropyl trimethyl ammonium chloride chitosan / N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle prepared by the invention, the particle size is easy to control, the particle size distribution is uniform, the maximum encapsulation rate can reach 98%, the medicine carrying quantity is large, and the compatibility and the biology adherence property are good. The preparation method is simple and easy to implement, the condition is mild, the toxic side effects of the medicine are fewer, the sustained-release time is long, the production cost is lower, and the large scale production is easy to implement.

Description

Technical field [0001] The invention relates to a preparation method of Newcastle disease live attenuated vaccine nanoparticles. Background technique [0002] Newcastle disease (ND) is an acute, highly contagious, and fatal poultry infectious disease caused by Newcastle disease virus (NDV). The disease occurs in chickens, turkeys, pigeons, cranes, quail and quail. Parrots and other birds can cause 100% morbidity and death. The main features of the disease are dyspnea, diarrhea, neurological disorders, mucosal and serous membrane hemorrhage, acute onset and high fatality rate. It is one of the most serious infectious diseases that endanger the poultry industry. At present, the most important and effective means of prevention and control of this disease is vaccination. However, the general disadvantage of existing vaccines is that they are weak in immunogenicity and require strong immune adjuvants or a suitable mucosal immune delivery system to effectively induce immune responses...

Claims

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Application Information

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IPC IPC(8): A61K9/19A61K39/17A61P31/14
Inventor 赵凯金政张旭陈刚吴昊
Owner HEILONGJIANG UNIV
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