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Application of chitosan oligosaccharide in preparing vaccine adjuvant and vaccine composition

A technology of vaccine composition and vaccine adjuvant, which is applied in the field of preparation of vaccine adjuvant and vaccine composition and vaccine preparation containing the vaccine adjuvant, can solve the problems of high price and application limitation, and achieve reasonable cost and controllable quality , easy absorption effect

Active Publication Date: 2019-04-02
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The very limited oligosaccharides that can be prepared at present are expensive, and their applications are greatly limited

Method used

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  • Application of chitosan oligosaccharide in preparing vaccine adjuvant and vaccine composition
  • Application of chitosan oligosaccharide in preparing vaccine adjuvant and vaccine composition
  • Application of chitosan oligosaccharide in preparing vaccine adjuvant and vaccine composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: preparation and physical and chemical properties of chitosan oligosaccharide

[0034] 1. Preparation of chitosan oligosaccharide:

[0035] Add deionized water into the batching tank, dissolve chitosan in 1% acetic acid solution, stir and heat up to 38±2.0°C. Add 5% (to the substrate) enzyme preparation, stir, and keep the temperature at 38°C. After the viscosity of the enzymolysis solution drops to 60%, start ultrafiltration (cut molecular weight at 6000). The part smaller than the cut-off molecular weight permeates the membrane, and the part larger than the cut-off molecular weight returns to the reactor to continue to degrade. The ultrafiltration permeate is passed through a nanomembrane filter (cutting molecular weight 300), and the permeated part is water, acetic acid and small molecule sugar (the permeated part can be recycled for preparing chitosan solution), and the non-permeated part is Concentrate into chitosan oligosaccharide solution and spr...

Embodiment 2

[0045] Embodiment 2: the adjuvant activity determination of chitosan oligosaccharide to influenza vaccine

[0046] The chitosan oligosaccharide in Example 1 was used as a vaccine adjuvant, and the H1N1 influenza vaccine (H1N1 influenza virus lysate, 30 μg / ml) was used as an antigen, and the two were injected intramuscularly to immunize mice, and the antibody titer was determined. The specific method is as follows:

[0047] Experimental animals: BALb / c mice, 6-8 weeks old, 5 mice / group, female.

[0048] Drug concentration: Chitooligosaccharide (COS) prepared in Example 1 above: 4 mg / ml and 20 mg / ml; H1N1 influenza virus lysate: 30 μg / ml; aluminum adjuvant (Al): 4 mg / ml.

[0049] Control solvent: physiological saline (Saline).

[0050] Drug concentration after mixing: chitosan oligosaccharide: 2 mg / ml and 10 mg / ml; H1N1 influenza virus lysate: 15 μg / ml; aluminum adjuvant (Al): 2 mg / ml.

[0051] Dosage: Oligochitosan: 200 μg / mouse (mouse) and 1 mg / mouse; H1N1: 1.5 μg / mouse; ...

Embodiment 3

[0066] Embodiment 3: the adjuvant activity of chitosan oligosaccharide to hepatitis B vaccine

[0067] The chitosan oligosaccharide in Example 1 was used as a vaccine adjuvant, and the recombinant hepatitis B subunit vaccine (HBsAg expressed by Hansenula yeast, produced by Dalian Hanxin Biomedical Co., Ltd.) was used as an antigen, and the two were used to immunize mice by intramuscular injection , Determination of antibody titers. Concrete experiment is with embodiment 2.

[0068] Dosage: Oligochitosan: 200 μg / mouse (mouse) and 50 μg / mouse; HBsAg: 2 μg / mouse; Aluminum adjuvant (Al): 200 μg / mouse.

[0069] Experimental grouping: (1) normal saline group (Saline); (2) aluminum adjuvant group (Al+HBsAg); (3) hepatitis B subunit group (Saline+HBsAg); (4) COS low-dose group: COS (200μg / mouse)+HBsAg; (5) COS high dose group: COS (1mg / mouse)+HBsAg.

[0070] Equal volumes were mixed before injection, 100 μl / mouse, and intramuscularly injected into the right hind limb.

[0071] ...

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Abstract

The invention discloses an application of chitosan oligosaccharide in preparing a vaccine adjuvant as well as a vaccine composition and a vaccine preparation containing the vaccine adjuvant. The vaccine adjuvant comprises chitosan oligosaccharide. The method for preparing the chitosan oligosaccharide of which the polymerization degree is 2-20 comprises the following steps: performing biological enzymatic hydrolysis, chemical decomposition or physical method decomposition on chitosan, wherein the deacetylation degree of the chitosan oligosaccharide of which the polymerization degree is 2-20 is greater than 90%, the molecular weight is 400-5000Da, and the sum of components of which the polymerization degrees are 3-10 is not less than 50%. The vaccine adjuvant containing chitosan can be used as an adjuvant of an attenuated vaccine, a nucleic acid vaccine, a polypeptide vaccine or protein vaccine.

Description

technical field [0001] The invention belongs to the technical field of medicine, and particularly relates to the use of chitosan oligosaccharide in preparing vaccine adjuvants and vaccine compositions and vaccine preparations containing the vaccine adjuvants. Background technique [0002] Adjuvant is a kind of substance that can enhance the immune effect of antigen when applied in advance of antigen or at the same time with antigen. Its main function is to enhance the specific immune response of the body's immune system to antigen. Since the British doctor Jenner vaccinated against smallpox more than 200 years ago and achieved success in human trials, humans can prevent and control certain infectious diseases through vaccination. In 1925, French immunologist and veterinary scientist Gaston Ramon discovered that adding tapioca starch, agar, pyogenic bacteria and other substances unrelated to antigens to diphtheria and tetanus vaccines could specifically enhance the body’s res...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K39/00A61K9/00
Inventor 杜昱光贾培媛单俊杰王玉霞许青松刘洪涛程功未金花
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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