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Vaccine adjuvant, vaccine composition, and use of sulfated chitooligosaccharide in preparation of vaccine adjuvant and vaccine composition

A vaccine composition and chitosan oligosaccharide technology, applied in the field of medicine, can solve the problems of serious adverse reactions of oil adjuvants, inflammation, ulcers and granulomas, and difficult metabolism of oil substances, and achieve good adjuvant effect and easy absorption , cost-effective effect

Inactive Publication Date: 2018-02-02
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

Oil adjuvant vaccines are superior to aluminum salt adjuvant vaccines in terms of antibody titer and immune persistence, but oil adjuvant has serious adverse reactions, and subcutaneous injection can cause inflammation, ulcers and granulomas; oily substances are not easy to remain in tissues for a long time. metabolism
[0003] With the deepening of immunology research and the rapid development of genetic engineering technology, various new genetic engineering vaccines have been developed for different diseases, and the purity of new technology vaccines such as epitope vaccines and genetic engineering vaccines is getting higher and higher. However, these vaccines generally have shortcomings such as small molecules, weak immunogenicity, and difficulty in inducing the body to produce an effective immune response. Therefore, immune adjuvants are needed to coordinate this effect, especially adjuvants that are safe and non-toxic, and can stimulate a strong cellular immune response. agents, and immune adjuvants suitable for mucosal vaccines, nucleic acid vaccines and tumor vaccines

Method used

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  • Vaccine adjuvant, vaccine composition, and use of sulfated chitooligosaccharide in preparation of vaccine adjuvant and vaccine composition
  • Vaccine adjuvant, vaccine composition, and use of sulfated chitooligosaccharide in preparation of vaccine adjuvant and vaccine composition
  • Vaccine adjuvant, vaccine composition, and use of sulfated chitooligosaccharide in preparation of vaccine adjuvant and vaccine composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: preparation and physical and chemical properties of chitosan oligosaccharide

[0041] 1. Preparation of chitosan oligosaccharides:

[0042] 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 sp...

Embodiment 2

[0053] Embodiment 2: Preparation and physicochemical properties of sulfated chitosan oligosaccharide

[0054] 1. Preparation of 6-O-sulfated chitosan oligosaccharide (COSS):

[0055] Concentrated sulfuric acid 20ml, HClSO3 10ml, chitosan oligosaccharide 500mg (504.3mg, 6.17mM), ice bath 30min, generate reddish-brown reaction solution, react at room temperature for 3h, absorb the reaction solution and add it to the Erlenmeyer flask equipped with pre-cooled ether solution, 4 Stand overnight at ℃, centrifuge at 10000rpm for 30min, wash the precipitate twice with 4℃ pre-cooled ether, centrifuge at 10000rpm for 30min, remove excess acid solution as much as possible, remove supernatant (add NaHCO3 powder to the supernatant solution to neutralize), and store in a fume hood Leave it to dry for 3 hours, dissolve the precipitate in 2ml of water, neutralize to neutral with 0.5M NaHCO3, and freeze-dry. The dried samples were desalted by Sephadex G15 column purification (Sephadex gel ch...

Embodiment 3

[0070] Example 3: Determination of adjuvant activity of sulfated oligochitosan to porcine reproductive and respiratory syndrome inactivated vaccine

[0071] The sulfated chitosan oligosaccharide in Example 2 was used as a vaccine adjuvant, and the inactivated porcine reproductive and respiratory syndrome vaccine (PRRSV) was used as an antigen, and the two were used in combination to immunize mice by intramuscular injection, and the antibody titer was determined. The specific method is as follows:

[0072] Experimental animals: BALb / c mice, 6-8 weeks old, 6 mice / group, female.

[0073] Drug concentration: oligochitosan (COS) prepared in the above-mentioned embodiment 1 and sulfated oligochitosan (COSS and SCOSS) prepared in embodiment 2: 4mg / ml; Porcine reproductive and respiratory syndrome inactivated vaccine (PRRSV , Guangdong Yongshun): 1.6mg / ml; ISA206 oil-emulsion adjuvant (French Seepic product): mixed with the vaccine at a volume ratio of 1:1.

[0074] Control solven...

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Abstract

The invention discloses a use of sulfated chitooligosaccharide in the preparation of a vaccine adjuvant, a vaccine composition containing the vaccine adjuvant and a vaccine preparation containing thevaccine adjuvant. The vaccine adjuvant comprises 6-O-sulfated chitooligosaccharide (COSS) and 2-N,6-O-sulfated chitooligosaccharide (SCOSS). Chitosan undergoes biological enzyme process hydrolysis, chemical process degradation or physical process cracking to generate chitooligosaccharide with the polymerization degree being 2-10 and the deacetylation degree being more than 90%, a chlorosulfonic acid-sulfuric acid method is used to carry out 6-O modification on the chitooligosaccharide to prepare the COSS, and a sulfonic acid-pyridine method is used to carry out sulfation modification on the amino group of the COSS to prepare the SCOSS. The molar weight range of the chitooligosaccharide with the polymerization degree being 2-10 in the sulfated chitooligosaccharide is 400-10000 Da, and the sulfur content (S%) of the chitooligosaccharide is more than 29%. The vaccine adjuvant containing the sulfated chitooligosaccharide can be used as an adjuvant for attenuated vaccines, inactivated vaccines, nucleic acid vaccines, polypeptide vaccines or protein vaccines.

Description

technical field [0001] The invention belongs to the technical field of medicine, in particular, the invention relates to the use of sulfated chitosan oligosaccharide in the preparation of 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 speci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39C08B37/08
CPCA61K39/39A61K2039/55511A61K2039/55583C08B37/003
Inventor 杜昱光贾培媛未金花张贵强程功焦思明冯翠
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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