Oligosaccharide vaccine for preventing invasive fungal infection

A technology for invasive fungi and vaccines, applied in the direction of antifungal agents, medical preparations with non-active ingredients, carrier-bound antigen/hapten components, etc., can solve the problem of lack of vaccines to prevent invasive fungal infections, and achieve broad-spectrum prevention Effects of fungal infection

Active Publication Date: 2019-03-08
SHANDONG ACADEMY OF PHARMACEUTICAL SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the current lack of vaccines for preventing invasive fungal infections, the present invention provid...

Method used

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  • Oligosaccharide vaccine for preventing invasive fungal infection
  • Oligosaccharide vaccine for preventing invasive fungal infection
  • Oligosaccharide vaccine for preventing invasive fungal infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Preparation of Chitooligosaccharides

[0033] 1.1

[0034] Dissolve 15g of chitosan with a degree of deacetylation of 1% in 300mL of 0.3M acetic acid, adjust the pH to 5.0 after the dissolution is complete, and add water to make the volume up to 500mL. Add 1.5g of chitosanase (130U / g) and stir in a water bath at 25-35°C for 8h. The chitosan enzymatic hydrolysate was filtered by plate and frame, and then ultrafiltration and nanofiltration with different pore size (400-5000Da) membranes were used to intercept the chitosan oligosaccharides ( 420 <0.1; 2% aqueous solution is centrifuged at 13000rpm for 2min without precipitation.

[0035] 1.2

[0036] Dissolve 18 g of chitosan with a degree of deacetylation of 95% in 300 mL of 0.2 M hydrochloric acid. After the dissolution is complete, adjust the pH to 6.5, and add water to make the volume to 500 mL. Add 1.5g of chitosanase (200U / g) and stir in a water bath at 25-35°C for 16h. The chitosan enzymatic hydrolysate is filt...

Embodiment 2

[0049] Example 2 Preparation of oligosaccharide vaccine

[0050] Take 50mg KLH dissolved in 1mL 0.1M sodium phosphate buffer pH7.2, containing 0.15M NaCl. Add 1 mg of chitosan with a degree of deacetylation of 10% and a molecular weight of 1000-3000Da, dissolve and mix. Take glutaraldehyde (CHO(CH 2 ) 3 CHO) 2μL of 50% aqueous solution, add the substrate solution, mix well, and keep it at 25°C with slight shaking for 60 minutes. Add 50mg of glycine solid and mix well, keep at 4℃ for 30min. Using 0.1M sodium phosphate buffer and 0.15M NaCl as the mobile phase, low molecular impurities were removed through Thermo Scientific™ Zeba™ SpinDesalting Columns to obtain the carrier protein solution coupled with chitosan oligosaccharide, which was placed in a 3000Da dialysis bag and dialyzed in pure water for 24h , Oligosaccharide vaccine is obtained after lyophilization.

Embodiment 3

[0051] Example 3 Preparation of oligosaccharide vaccine

[0052] Take 50mg KLH dissolved in 1mL 0.1M sodium phosphate buffer pH7.2, containing 0.15M NaCl. Add 1 mg of chitosan with a degree of deacetylation of 95% and a molecular weight of 1000-3000Da, dissolve and mix. Take glutaraldehyde (CHO(CH 2 ) 3 CHO) 11μL of 50% aqueous solution, add the substrate solution, mix well, and keep it at 25°C and shake it slightly for 60 minutes. Add 250mg of glycine solid and mix well, keep at 4℃ for 30min. Using 0.1M sodium phosphate buffer, containing 0.15M NaCl as the mobile phase, remove low-molecular impurities through Thermo Scientific™ Zeba™ SpinDesalting Columns to obtain the carrier protein solution coupled with chitosan oligosaccharide, and place it in a 2000Da dialysis bag with pure water for 24h dialysis , Oligosaccharide vaccine is obtained after lyophilization.

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Abstract

The invention provides an oligosaccharide vaccine for preventing invasive fungal infection. The oligosaccharide vaccine comprises chitosan oligosaccharide coupled with an immunogenic carrier protein;the mass ratio of the immunogenic carrier protein to the chitosan oligosaccharide is 0.7-210: 1; the chitosan oligosaccharide has a deacetylation degree of 0.05-98% and an average relative molecular weight smaller than 5000 Da; and the immunogenic carrier protein is a non-human protein. The oligosaccharide vaccine provided by the invention is capable of effectively activating Th17 and Th1 cellularimmunity through a proper acetylation degree so as to prevent broad-spectrum fungal infections. The oligosaccharide vaccine is capable of treating or preventing invasive fungi such as candida albicans, aspergillus, cryptococcus and pneumocystis jeroveci.

Description

Technical field [0001] The invention belongs to the field of vaccine biomedicine, and specifically relates to an oligosaccharide vaccine for preventing invasive fungal infection. Background technique [0002] With the increase in the incidence of new immunosuppressive treatments, organ transplantation and AIDS, the incidence and mortality of invasive drug-resistant fungal infections have increased significantly worldwide. According to international medical standards, fungal infections are divided into three types: superficial, subcutaneous tissue, and invasive (or systemic) fungal infections. The latter two are also called deep fungal infections. The common pathogenic strains are Candida, Aspergillus, Cryptococcus and Pneumocystis jiroviz, which account for about 70% of fungal infections. The infection involves internal organs or forms fungalemia. Clinical deep fungal infections have become an important cause of death from hospital infections, and 1.5 million people die from fun...

Claims

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Application Information

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IPC IPC(8): A61K39/385A61K47/64A61P31/10
CPCA61K39/385A61K47/64A61P31/10
Inventor 陈勉刘飞凌沛学孙康陈磊袁丹丹张小刚张天娇张秀华张金华袁超张林军刘霞刘英梅
Owner SHANDONG ACADEMY OF PHARMACEUTICAL SCIENCES
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