Development of cold - chain free oral protein-based drug

• Project/Area Number: 21200073
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research a proposed research project)
• Allocation Type: Single-year Grants
• Research Field: Applied molecular and cellular biology; Applied pharmacology
• Research Institution: The University of Tokyo
• Principal Investigator: YOSHIKAZU Yuki 東京大学, 医科学研究所, 助教 (60345030)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥30,940,000 (Direct Cost: ¥23,800,000、Indirect Cost: ¥7,140,000); Fiscal Year 2011: ¥10,010,000 (Direct Cost: ¥7,700,000、Indirect Cost: ¥2,310,000); Fiscal Year 2010: ¥10,010,000 (Direct Cost: ¥7,700,000、Indirect Cost: ¥2,310,000); Fiscal Year 2009: ¥10,920,000 (Direct Cost: ¥8,400,000、Indirect Cost: ¥2,520,000)
• Keywords: オルガネラ工学 / ドラッグデリバリー / 経口蛋白医薬 / コメ / RNAi / ドラックデリバリー / 医療福祉 / バイオテクノロジー / 免疫学 / 薬学

Research Abstract

We have developed a novel system for the production of protein drug such as IL-10 and rotavirus-specific llama heavy-chain antibody (VHH) using rice-based expression system (MucoRice). Orally administered MucoRice-VHH markedly decreased the viral load in immunocompetent as well as immunodeficient mice. MucoRice-ARP1 retained in vitro neutralizing activity after long-term storage at RT and boiling. High-yield, water-soluble and purification-free MucoRice-VHH thus forms the basis for orally administered therapy against rotavirus infections.

Research Products

• [Journal Article] Nanogel-based PspA intranasal vaccine prevents invasive disease and nasal colonization by Pneumococcus (2013)
  • Author(s): IG. Kong, A. Sato, Y. Yuki, T. Nochi, H. Takahashi, S. Sawada, M. Mejima, S. Kurokawa, K. Okada, S. Sato, D. Briles, J. Kunisawa, Y. Inoue, M. Yamamoto, K. Akiyoshi, and H. Kiyono
  • Journal Title: Infect. & Immun
  • Peer Reviewed
• [Journal Article] Induction of Toxin-specific Neutralizing Immunity by Molecularly Uniform Rice-based Oral Cholera Toxin B Subunit Vaccine without Plant-associated Sugar Modification (2013)
  • Author(s): Y. Yuki, M. Mejima, S. Kurokawa, T. Hiroiwa, Y. Takahashi, D. Tokuhara, T. Nochi, Y. Katakai, M. Kuroda, N. Takeyama, K. Kashima, M. Abe, Y. Chen, U. Nakanishi, T. Masumura, Y. Takeuchi, H. Kozuka-Hata, H. Shibata, M. Oyam|, K. Tanaka, H. Kiyono
  • Journal Title: Plant Biotechnol
  • Peer Reviewed
• [Journal Article] Figihashi. Potential roles of CCR5+CCR6+ dendric cells induced by nasal ovalbumin plus Flt3 ligand expressing adnovirus for mucosal IgA responses (2013)
  • Author(s): Y. Fukiyama, D. Tokuhara, S. Sekine, K. Aso, K. Kataoka, J. Davydova, M. Yamamoto, RS Gilbert, Y. Tokuhara, K. Fujihashi, J. Kunisawa, Y. Yuki, H. Kiyono, JR McGhee, K
  • Journal Title: PloS One Volume: 8
  • Peer Reviewed
• [Journal Article] RNAi suppression of rice endogenous storage proteins enhances the production of rice-based Botulinum neutrotoxin type A vaccine (2012)
  • Author(s): Y. Yuki, M. Mejima, S. Kurokawa, T. Hiroiwa, IG. Kong, M. Kuroda, Y. Takahashi, T. Nochi, D. Tokuhara, T. Kohda, S. Kozaki, H. Kiyono
  • Journal Title: Vaccine Volume: 30 Pages: 4160-4166
  • Peer Reviewed
• [Journal Article] Transcription factor Spi-B-dependent and -independentpathways for the development of Peyer's patch M cells (2012)
  • Author(s): S. Sato, S. Kaneto, N. Shibata, Y. Takahashi, H. Okura, Y. Yuki, J. Kunisawaand H. Kiyono
  • Journal Title: Mucosal Immunol
  • Peer Reviewed
• [Journal Article] Novel vaccine development strategies for inducing mucosal immunity (2012)
  • Author(s): Fujkuyama Y, Tokuhara D, Kataoka K, Gilbert RS, McGhee JR, Yuki Y, Kiyono H, Fujihashi K
  • Journal Title: Expert Rev. Vaccines Volume: 11 Pages: 376-379
  • Peer Reviewed
• [Journal Article] Transcription factor Spi-B-dependent and -independentpathways for the development of Peyer' s patch M cells. (2012)
  • Author(s): S. Sato, S. Kaneto, N. Shibata, Y. Takahashi, H. Okura, Y. Yuki., et. al.
  • Journal Title: Mucosal Immunol. Volume: Dec. 5(in press)
  • Peer Reviewed
• [Journal Article] Distinct fucosylation of M cells and epithelial cells by Fut1 and Fut2, respectively, in response to intestinal environmental stress (2011)
  • Author(s): K. Terahara, T. Nochi, M. Yoshida, Y. Takahashi, Y. Goto, H. Hatai, S. Kurokawa, M-H. Jang, M-N. Kweon, SE. Domino, T. Hiroi, Y. Yuki, Y. Tsunetsugu-Yokota, K. Kobayashi, H. Kiyono
  • Journal Title: Biochem. Biophys. Res. Commun Volume: 404 Pages: 822-828
  • Peer Reviewed
• [Journal Article] Indigenous opportunistic bacteria inhabit mammalian gut-associated lymphoid tissues and share a mucosal antibody-mediated symbiosis (2010)
  • Author(s): T. Obata, Y. Goto, J. Kunisawa, S. Sato, M. Sakamoto, H. Setoyama, T. Matsuki, K. Nonaka, N. Shibata, M. Gohda, Y. Kagiyama, T. Nochi, Y. Yuki, Y. Fukuyama, A. Mukai, S. Shinzaki, K. Fujihashi, C. Sasakawa, H. Iijima, M. Goto, Y. Umesaki, Y. Benno, H. Kiyono
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 107 Pages: 7419-7424
  • Peer Reviewed
• [Journal Article] Secretory IgA-mediated protection against V. cholerae and heat-labile enterotoxin-producing enterotoxigenic Escherichia coli by rice-based vaccine (2010)
  • Author(s): D. Tokuhara, Y. Yuki, T. Nochi, T. Kodama, M. Mejima, S. Kurokawa, Y. Takahashi, M. Nanno, U. Nakanishi, F. Takaiwa, T. Honda, H. Kiyono
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 107 Pages: 8794-8799
  • Peer Reviewed
• [Journal Article] Nanogel antigenic protein-delivery system for adjuvant-free intranasal vaccines (2010)
  • Author(s): T. Nochi, Y. Yuki, H. Takahashi, S. Sawada, M. Mejima, T. Kohda, N. Harada, IG. Kong, A. Sato, N. Kataoka, D. Tokuhara, S. Kurokawa, Y. Takahashi, H. Tsukada, S. Kozaki, K. Akiyoshi, H. Kiyono
  • Journal Title: Nat. Materials Volume: 9 Pages: 572-578
  • Peer Reviewed
• [Journal Article] In vivo molecular imaging analysis of a nasal vaccine that induces protective immunity against botulism in non-human primates (2010)
  • Author(s): Y. Yuki, T. Nochi, N. Harada, Y. Katakai, H. Shibata, M. Mejima, T. Kohda, D. Tokuhara, S. Kurokawa, Y. Takahashi, F. Ono, S. Kozaki, K. Terao, H. Tsukada, H. Kiyono
  • Journal Title: J. Immunol Volume: 185 Pages: 5436-5443
  • Peer Reviewed
• [Journal Article] Secretory IgA-mediated protection against V.cholerae and heat-labile enterotoxin-producing enterotoxigenic Escherichia coli by rice-based vaccine. (2010)
  • Author(s): D.Tokuhara, Y.Yuki, et al
  • Journal Title: Proc.Natl.Acad.Sci.USA. Volume: 107 Pages: 8794-8799
  • Peer Reviewed
• [Journal Article] Nanogel antigenic protein-delivery system for adjuvant-free intranasal vaccines. (2010)
  • Author(s): Nochi T, Yuki Y, et al.
  • Journal Title: Nat.Mater. Volume: 9 Pages: 572-578
  • Peer Reviewed
• [Journal Article] In vivo molecular imaging analysis of a nasal vaccine that induces protective immunity against botulism in non-human primates. (2010)
  • Author(s): Yuki Y, Nochi T, et al.
  • Journal Title: J.Immunol. Volume: 185 Pages: 5436-5443
  • Peer Reviewed
• [Journal Article] A rice-based oral cholera vaccine induces macaque-specific systemic neutralizing Abs but does not influence pre-existing intestinal immunity (2009)
  • Author(s): Nochi, T., Yuki, Y., et.al.
  • Journal Title: J.Immunol. 183 Pages: 6538-6544
  • Peer Reviewed
• [Journal Article] Oral MucoRice expressing double-mutant cholera toxin A and B subunits induces toxin-specific neutralizing immunity (2009)
  • Author(s): Yuki, Y., Tokuhara, D., et.al.
  • Journal Title: Vaccine 27 Pages: 5982-5988
  • Peer Reviewed
• [Presentation] Heat tolerance evaluation of the rice type oral cholera vaccine, MucoRice-CTB, for a clinical study (2012)
  • Author(s): K. Kashima, H. Hiroiwa, Y. Yuki, H. Kiyono.
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 東京
  • Year and Date: 2012-11-17
• [Presentation] MucoRice-CTB as an oral vaccine for the prevention of entroxigenic E. coli-mediated diarrhea in pigs. (2012)
  • Author(s): Takeyama, K. Oroku, D. Tokuhara, S. Nagai, H. Kiyono, Y. Yuki
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 東京
  • Year and Date: 2012-11-17
• [Presentation] Characterization of effector/memory CD4 T cells expanded in the mice vaccinated with MucoRice-CTB (2012)
  • Author(s): EJ. Park, S. Joo, S. Kurokawa, H. Kiyono, Y. Yuki
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 東京
  • Year and Date: 2012-11-17
• [Presentation] ブタ大腸菌性下痢症予防食べるワクチンの開発研究 (2012)
  • Author(s): 幸義和
  • Organizer: 動物ワクチン-バイオ医薬研究会
  • Place of Presentation: 盛岡(招待講演)
  • Year and Date: 2012-09-04
• [Presentation] コメ型経口ワクチンMucoRice-CTBの研究開発 (2012)
  • Author(s): 幸義和
  • Organizer: 日本薬学会
  • Place of Presentation: 札幌(招待講演)
  • Year and Date: 2012-03-29
• [Presentation] In vivo molecular imaging analysis of a nasal vaccine. (2011)
  • Author(s): Y.Yuki, et.al.
  • Organizer: 3rd World Congress of Vaccine
  • Place of Presentation: Beijing, China
  • Year and Date: 2011-03-24
• [Presentation] Rice-based oral vaccine of cholera toxin B subunit for the protection of enteroxigenic Escherichia coli infection in pigs (2011)
  • Author(s): N. Takayama, D. Tokuhara, T. Nochi, K. Oroku, S. Nagai, A. Chubachi, K. Tuchiya, Y. Yuki & H. Kiyono
  • Organizer: 3rd World Congress of Vaccine
  • Place of Presentation: Beijing, China
• [Presentation] In vivo molecular imaging for a nasal botulism vaccine in mice and nonhuman primates (2011)
  • Author(s): Y. Yuki, T. Nochi, N. Harada, Y. Katakai, M. Mejima, D. Tokuhara, S. Kurokawa, Y. Takahashi, H. Shibata, T. Kohda, S. Kozaki, H. Tsukada, & H. Kiyono
  • Organizer: The American Association of Immunologists (AAI)
  • Place of Presentation: San Francisco, USA
• [Presentation] Passive oral rice-based antibody prophylaxis and therapy against rotavirus infection (2011)
  • Author(s): D. Tokuhara, B. Gonzalez, M. Mejima, Y. Takahashi, S. Kurokawa, T. Hiroiwa, M. Kuroda, M. Oyama, H. Kozuka-Hata, T. Nochi, F. Aladin, H. Marcotte, L. Frenken, M. Iturriza, H. Kiyono, L. Hammarstrom, & Y. Yuki
  • Organizer: The American Association of Immunologists (AAI)
  • Place of Presentation: San Francisco, USA
• [Presentation] Nanogel based adjuvant-free Pneumococcal nasal vaccine induces protective immunity against Pneumococcus (2011)
  • Author(s): IG. Kong, Y. Yuki, A. Sato-Kimura, T. Nochi, H. Takahashi, S. Sawada, M. Mejima, S. Kurokawa, D. Tokuhara, K. Okada, S. Sato, D. Briles, K. Akiyoshi & H. Kiyono
  • Organizer: International congressof Mucosal Immunology (ICMI)
  • Place of Presentation: Paris, France
• [Presentation] Indispensableness of vaccine -delivery to Peyer's patch and villous M cells for the induction of antigen-specific immune response (2011)
  • Author(s): GS. Pontes, T. Nochi, H. Nakamura-Adachi, Y. Kurashima, J. Kunisawa, Y. Yuki & H. Kiyono
  • Organizer: International congressof Mucosal Immunology (ICMI)
  • Place of Presentation: Paris, France
• [Presentation] In vivo molecular imaging for a nasal botulism vaccine in mice and nonhuman primates (2011)
  • Author(s): Y. Yuki, T. Nochi, N. Harada, Y. Katakai, H. Tsukada, & H. Kiyono
  • Organizer: Vaccine and ISV Annual Global Congress
  • Place of Presentation: Seattle, USA
• [Presentation] L. Hammarstrom. Rice-based oral antibody prophylaxis and therapy against rotavirus infection (2011)
  • Author(s): Y. Yuki, D. Tokuhara, B. A. Gonzalez, H. Marcotte, L. Frenken, M. Iturriza, H. Kiyono
  • Organizer: 日本免疫学会
  • Place of Presentation: 千葉
• [Presentation] 経鼻ワクチンの安全性:マウスおよびサルに経鼻投与されたワクチンのリアルタイム分子イメージング (2011)
  • Author(s): 幸 義和、清野 宏
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 東京
• [Presentation] アジュバント不要ナノゲル型経鼻ワクチンの開発 (2010)
  • Author(s): 幸義和, ら
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 東京 九段会館
  • Year and Date: 2010-12-11
• [Presentation] Exploit of Nanogel-based Delivery System for Adjuvant-free Intranasal Vaccines. 8th Annual Vaccines (2010)
  • Author(s): Y.Yuki
  • Organizer: 8th Annual Vaccines : All Things Considered Conference
  • Place of Presentation: Washington, DC USA(招待講演)
  • Year and Date: 2010-11-09
• [Presentation] Exploit of Novel Delivery System with Nanogel for Development of Adjuvant-Free Nasal Vaccine. (2010)
  • Author(s): Y.Yuki, et.al.
  • Organizer: International conference of Immunology
  • Place of Presentation: Kobe, Japan 神戸国際展示場
  • Year and Date: 2010-08-25
• [Presentation] A rice-based oral cholera vaccine induces macaque-specific systemic neutralizing Abs but does not influence natural intestinal immunity (2010)
  • Author(s): Y.Yuki, et.al.
  • Organizer: The American Association of Immunologists
  • Place of Presentation: Baltimore, USA
  • Year and Date: 2010-05-10
• [Presentation] A Novel Nanogel-based Delivery System for Adjuvant-free Intranasal Vaccines (2010)
  • Author(s): Y. Yuki
  • Organizer: 2nd World Congress of Vaccine
  • Place of Presentation: Beijing, China
• [Presentation] A rice-based oral cholera vaccine induces macaque-specific systemic neutralizing Abs but does not influence natural intestinal immunity (2010)
  • Author(s): Y. Yuki, T. Nochi, Y. Katakai, H. Shibata, D. Tokuhara, M. Mejima, S. Kurokawa, Y. Takahashi, H. Hatai, A. Chubachi, U. Nakanishi, K. Terao, H. Kiyono
  • Place of Presentation: Baltimore, USA
• [Presentation] Nanogel antigen delivery system for adjuvant-free intranasal vaccines (2010)
  • Author(s): T. Nochi, Y. Yuki, H. Takahashi, S. Sawada, M. Mejima, T. Kohda, N. Harada, N. Kataoka, IG. Kong, A. Sato, D. Tokuhara, S. Kurokawa, Y. Takahashi, H. Tsukada, S. Kozaki, K. Akiyoshi, H. Kiyono
  • Place of Presentation: Baltimore, USA
• [Presentation] Secretory IgA responses induced by rice-based oral cholera toxin B subunit vaccine are solely responsible for antibody-mediated long-standing cross-protection against Vibrio cholerae- and enterotoxigenic Escherichia coli- induced diarrhea (2010)
  • Author(s): D. Tokuhara, Y. Yuki, T. Nochi, T. Kodama, M. Mejima, S. Kurokawa, Y. Takahashi, M. Nanno, F. Takaiwa, T. Honda, H. Kiyono
  • Place of Presentation: Baltimore, USA
• [Presentation] Exploit of Novel Delivery System with Nanogel for Development of Adjuvant-Free Nasal Vaccine (2010)
  • Author(s): Y. Yuki, T. Nochi, H. Takahashi, S. Sawada, M. Mejima, T. Kohda, N. Harada, N. Kataoka, IG. Kong, A. Sato, D. Tokuhara, S. Kurokawa, Y. Takahashi, H. Tsukada, S. Kozaki, K. Akiyoshi, H. Kiyono
  • Organizer: International conference of Immunology
  • Place of Presentation: Kobe. Japan
• [Presentation] Exploit of Nanogel-based Delivery System for Adjuvant-free Intranasal Vaccines (2010)
  • Author(s): Y. Yuki
  • Organizer: 8th Annual Vaccines: All Things Considered Conference
  • Place of Presentation: Washington, DC USA
• [Presentation] 米型コレラ毒素B鎖ワクチンは分泌型IgAを誘導することでコレラ菌による下痢を長期間防御する (2010)
  • Author(s): 幸 義和、徳原大介、野地 智法、児玉年央、目島未央、黒河志保、高橋裕子、南野昌信、本田 武司、清野 宏
  • Organizer: 日本細菌学会
  • Place of Presentation: 横浜
• [Presentation] アジュバント不要ナノゲル型経鼻ワクチンの開発 (2010)
  • Author(s): 幸 義和、野地智法、秋吉一成、清野 宏
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 東京
• [Presentation] In vivo molecular imaging analysis of a nasal vaccine (2010)
  • Author(s): Y. Yuki, T. Nochi, N. Harada, Y. Katakai, H. Shibata, M. Mejima, T. Kohda, D. Tokuhara, S. Kurokawa, Y. Takahashi, F. Ono, S. kazaki, K. Terao, H. Tsukada, H. Kiyono
  • Organizer: 3rd World Congress of Vaccine
  • Place of Presentation: Beijing, China
• [Presentation] Oral rice-based vaccine induced active and passive immunity against E. coli-mediated diarrhea in pigs (2010)
  • Author(s): N. Takeyama, D. Tokuhara, K. Oroku, M. Mijima, S. Kurokawa, Y. Takahashi, M. Kuroda, T. Hiroiwa, YG, Chen, A. Chubachi, T. Nochi, T. Masumura, K. Tanaka, T. Kodama, S. Nagai, T. Nunoya, H. Kiyono & Y. Yuki
  • Organizer: International congressof Mucosal Immunology (ICMI)
  • Place of Presentation: Paris, France
• [Presentation] An efficient rice-based expression system producing nasally immunogenic non-toxic subunit fragment of Clostridium Botulinum type-A neutrotoxin (2010)
  • Author(s): Y. Yuki, M. Mejima, M. Kuroda, S. Kurokawa, T. Hiroiwa, IG kong, Y. Takahashi, YG, Chen, T. Nochi, D. Tokuhara, T. Kohda, S. Kozaki & H. Kiyono
  • Organizer: International congressof Mucosal Immunology (ICMI)
  • Place of Presentation: Paris, France
• [Presentation] Development of a cold-chain free Rice-based oral vaccine : MucoRice (2009)
  • Author(s): 幸義和
  • Organizer: International collabotarion in Biotechnology : Research and Industry
  • Place of Presentation: osaka
  • Year and Date: 2009-10-09
• [Presentation] コレラ毒素Bサブユニット(CTB)発現米はカニクイザルへの経口投与により血清中にCT特異的IgG抗体を誘導するが、自然抗体として腸管に存在するCT特異的SIgA抗体の免疫応答には影響を与えない (2009)
  • Author(s): 幸 義和、野地智法、徳原大介、清野 宏
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 札幌
• [Presentation] コレラ毒素Bサブユニット発現コメ型経口ワクチンによるコレラ菌および毒素原性大腸菌に対する下痢予防効果の検証 (2009)
  • Author(s): 徳原大介、幸 義和、野地智法、清野 宏
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 札幌
• [Book] ワクチンのDDS技術の開発動向-経鼻投与技術動向- 「ワクチンの市場動向と開発・製造実務集」 (2012)
  • Author(s): 幸 義和
  • Publisher: 技術情報協会
• [Book] ワクチンのDDS技術の開発動向-経口投与技術動向- 「ワクチンの市場動向と開発・製造実務集」 (2012)
  • Author(s): 幸 義和
  • Publisher: 技術情報協会
• [Book] ワクチンの市場動向と開発・製造実務集 (2012)
  • Author(s): 幸義和, ら多数
  • Publisher: 技術情報協会
• [Book] 臨床粘膜免疫学(経口ワクチン、p532-539) (2010)
  • Author(s): 幸義和(清野宏編)
  • Total Pages: 722
  • Publisher: (株)シナジー
• [Book] Allergy Frontiers : Future Perspectives(Transgenic Rice) (2009)
  • Author(s): Y.Yuki, F.Takaiwa, H.Kiyono
  • Publisher: Springer
• [Remarks]
  • URL: http://www.ims.u-tokyo.ac.jp/imsut/jp/research/papers/post_21.php
• [Patent(Industrial Property Rights)] ナノ抗体を発現するトランスジェニックイネ (2010)
  • Inventor(s): 幸 義和、徳原大介、清野 宏
  • Industrial Property Rights Holder: 東京大学TLO
  • Industrial Property Number: 2010-162630
  • Filing Date: 2010-07-20
• [Patent(Industrial Property Rights)] 特許 (2010)
  • Inventor(s): 幸義和、徳原大介、清野宏
  • Industrial Property Rights Holder: 東大TLO
  • Industrial Property Number: 2010-162630
  • Filing Date: 2010-07-20