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Mucosal vaccine using cationic nanogel

a technology of cationic nanogels and mucosal vaccines, applied in the field of mucosal vaccines, can solve problems such as the safety of mucosal vaccines on organisms, and achieve the effects of indubiting systemic and mucosal immune responses, facilitating immune response, and facilitating immune respons

Inactive Publication Date: 2011-08-25
NAT UNIV CORP TOKYO MEDICAL & DENTAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"The present invention provides a mucosal vaccine that can be administered through the nasal or oral cavity without the need for a mucosal adjuvant like a toxin-related protein. The vaccine is made up of a nanogel that can be easily mixed with a vaccine antigen and is effective in inducing immune responses in organisms. The nanogel can be derived from a hydrophilic polysaccharide with hydrophobic cholesterol added thereto as a side chain. The vaccine can be used to prevent or treat infections caused by viruses, bacteria, protozoans, and fungi. The method for producing the vaccine involves mixing the nanogel with the vaccine antigen at room temperature for a few hours."

Problems solved by technology

However, such mucosal adjuvant may disadvantageously migrate to the brain, and the safety thereof on organisms has remained problematic.

Method used

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  • Mucosal vaccine using cationic nanogel
  • Mucosal vaccine using cationic nanogel
  • Mucosal vaccine using cationic nanogel

Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of Mucosal Vaccine

[0056]Cationic nanogels (cationic CHP) in which the degree of cholesterol substitution was 1.4 and the degree of ethylenediamine substitution was 18 per 100 monosaccharides were used (CHPNH2 nanogels). A CHP derivative or cationic Pullulan was dissolved in a 1 mg / ml phosphate buffer solution (PBS). The CHPNH2 nanogels were subjected to sonication for 15 minutes and then filtered through a 0.22-mm filter.

[0057]The C-terminal avirulent region of a heavy chain of botulinus toxin (Hc; molecular weight: 45,000), tetanus toxoid (TT; molecular weight: 150,000), or the AIDS virus membrane antigen molecule (gag p24; molecular weight: 24,000) expressed in E. coli and purified was mixed with the equimolar amount of cationic nanogels prepared in the manner described above, and the resulting mixture was subjected to reaction at 45° C. for 5 hours to prepare a composite. The obtained antigen / cationic nanogel composite was used as a mucosal vaccine using cationic nano...

example 2

Transnasal Immunization

[0058]The mucosal vaccine using cationic nanogels prepared in Example 1 or the antigen alone was administered to 6- to 8-week-old Balb / c mice (female) through the nasal cavity in an amount of 10 μg of Hc (88.9 μg of nanogel), 30 μg of TT (80.0 μg of nanogel), or 10 μg of gag p24 (166.7 μg of nanogel) per mouse once a week (3 times in total) to immunize mice transnasally. The amount of antigens administered (i.e., the amount of the solution) was adjusted to 15 μl in every experimental group, and 7.5 μl of the solution was administered to each nostril. PBS was administered as a control.

[0059]The blood was sampled before immunization and a week after immunization, and IgG antibody titers to botulinus toxin, TT, or gag p24 in the blood serum were measured to evaluate the systemic immune responses. The nasal cavity was washed with 200 μl of PBS a week after the final immunization, and the IgA antibody titer in the nasal wash solution was measured to evaluate immune...

example 3

Neutralization Effects after Transnasal Immunization using Mucosal Vaccine using Nanogels

[0065]The vaccine using cationic nanogels using a C-terminal avirulent region of the heavy chain of botulinus toxin (Hc; molecular weight: 45,000) as the antigen prepared in Example 1 or Hc alone was administered transnasally to 5 mice for immunization in the same manner as in Example 2. PBS was administered as a negative control. After the mice were subjected to immunization 3 times, botulinus toxin (obtained from Professor Shunji Kozaki, Division of Veterinary Science, School of Life and Environmental Sciences, Osaka Prefecture University) was administered intraperitoneally in an amount 25,000 times greater than the lethal dose thereof via intraperitoneal administration (i.g., 500 ng) to analyze the survival effects. For the purpose of analyzing the neutralization effects of Hc-specific IgA induced in the nasal tissue, 10 μg of botulinum progenitor toxins (obtained from Wako Pure Chemical Indu...

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Abstract

A mucosal vaccine for the prevention or treatment of microbial infections is described that is capable of inducing vaccine antigen-specific immune responses in an organism without the addition of a mucosal adjuvant. The mucosal vaccine comprises a composite of a nanogel comprising a hydrophilic polysaccharide having a cationic functional group and a hydrophobic cholesterol added thereto as a side chain and a vaccine antigen. The vaccine is administered via a mucosal route.

Description

TECHNICAL FIELD[0001]The present invention relates to a mucosal vaccine comprising a composite of a vaccine antigen and a cationic nanogel that is transnasally or orally administered.BACKGROUND ART[0002]Non-injection mucosal vaccines are safe and convenient to use, and thus they have drawn attention as the next-generation vaccines. It was necessary to administer a mucosal vaccine simultaneously with a mucosal adjuvant in order to induce effective antigen-specific immune responses with the use of a mucosal vaccine. As mucosal adjuvants, toxin-related proteins, such as cholera toxins (CT) or detoxicated cholera toxins (mCT), are known. Addition of such mucosal adjuvants to mucosal vaccines enables transnasal vaccines to induce mucosal IgA in addition to antigen-specific systemic IgG. However, such mucosal adjuvant may disadvantageously migrate to the brain, and the safety thereof on organisms has remained problematic.[0003]The present inventors developed nanogels comprising molecules ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/12A61K39/00A61K39/02A61K39/002A61P37/00
CPCA61K9/0043A61K9/006A61K9/06A61K9/5161A61K39/39A61K47/36A61K2039/6087A61K47/4823A61K47/48784A61K2039/543A61K2039/55583A61K39/08A61K2039/541A61K47/48123A61K47/554A61K47/61A61K47/6903A61P31/00A61P31/04A61P31/10A61P31/12A61P31/18A61P33/00A61P33/02A61P37/00A61P37/04
Inventor AKIYOSHI, KAZUNARIKIYONO, HIROSHIYUKI, YOSHIKAZUNOCHI, TOMONORI
Owner NAT UNIV CORP TOKYO MEDICAL & DENTAL UNIV
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