Ｃｌａｕｄｉｎを標的とした経皮吸収促進技術の開発

2014 Fiscal Year Annual Research Report

• Project/Area Number: 24390042
• Research Institution: Osaka University
• Principal Investigator: 近藤 昌夫 大阪大学, 薬学研究科(研究院), 准教授 (50309697)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: Claudin / Tight junction / 上皮バリア / 非侵襲性投与

Outline of Annual Research Achievements

近年､ペプチド･蛋白質･核酸医薬品等の画期的バイオ医薬シーズが創製されつつあるものの､バイオ医薬は生体膜透過性に乏しい上に消化酵素によって分解されやすく､多くのバイオ医薬が注射による投与を余儀なくされている｡周知のように、経皮投与は理想的な投与方法として注目されているものの､元来､皮膚は外部環境から生体内部環境を保護するバリアとして機能しており､ここに経皮投与薬創製の難しさがある｡本研究では､独自の生体バリア制御研究基盤を有効活用することで､皮膚バリア制御分子（Claudin-1 (CL-1) binder）を創製し､経皮投与基盤技術の開発を目指した｡
CL発現系の開発がCL binderスクリーニングの最重要課題となっていたが､現在までにバキュロウイルス（BV）上にCLが高密度提示されること､CL提示BVがCL binderスクリーニング系として機能することを見出している｡さらに､ファージ表面提示法を駆使することでCL-1 binderを創製できる可能性を見出してきた
これらの予備検討の結果を有効活用し、計画1年目はCL提示BVシステム、ファージ表面提示法、FRETシステムなどを活用したCL-1 binder創製システムの確立・最適化を実施した。計画2年目は、初年度に確立・最適化したCL-1 binder創製システムを有効活用することで、CL-1 binderスクリーニング系のシステムアップを図り、CL-1の細胞外領域認識抗体を創製した。3年目は、抗CL-1抗体の皮膚バリア制御活性解析に研究資源を集中し、抗CL-1抗体はヒト腸管バリアモデルには作用せず、ヒト皮膚バリアモデルにおいてのみバリア低下作用を示すことを見出し、CL-1を標的とした経皮吸収促進剤のコンセプトを検証した。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Causes of Carryover

26年度が最終年度であるため、記入しない。

Expenditure Plan for Carryover Budget

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Homoharringtonine increases intestinal epithelial permeability by modulating specific claudin isoforms in Caco-2 cell monolayers (2015)
  • Author(s): Akihiro Watari, Maki Hasegawa, Kiyohito Yagi, Masuo Kondoh
  • Journal Title: Eur J Pharm Biopharm Volume: 89 Pages: 232-238
  • DOI: 10.1016/j.ejpb.2014.12.012
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Discovery of anti-claudin-1 antibodies as candidate therapeutics against hepatitis C virus (2015)
  • Author(s): Mayo Yamashita, Manami Iida, Minoru Tada, Yohitaka Shirasago, Masayoshi Fukasawa, Shotaro Nagase, Akihiro Watari, Akiko Ishii-Watabe, Kiyohito Yagi, Masuo Kondoh
  • Journal Title: J Pharmacol Exp Ther Volume: 353 Pages: 112-118
  • DOI: 10.1124/jpet.114.217653
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Monoclonal antibodies against extracellular domains of claudin-1 block hepatitis C virus infection in a mouse model (2015)
  • Author(s): Masayoshi Fukasawa, Shotaro Nagase, Yoshitaka Shirasago, Manami Iida, Mayo Yamashita, Kohki Endo, Kiyohito Yagi, Tetsuro Suzuki, Takaji Wakita, Kentaro Hanada, Hiroki Kuniyasu, Masuo Kondoh
  • Journal Title: J Virol Volume: 89 Pages: 4866-4879
  • DOI: 10.1128/JVI.03676-14
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Tissue distribution and safety evaluation of a claudin-targeting molecule, the C-terminal fragment of Clostridium perfringens enterotoxin (2014)
  • Author(s): Xiangru Li, Rie Saeki, Akihiro Watari, Kiyohito Yagi, Masuo Kondoh
  • Journal Title: Tissue distribution and safety evaluation of a claudin-targeting molecule, the C-terminal fragment of Clostridium perfringens enterotoxin Volume: 52 Pages: 132-137
  • DOI: 10.1016/j.ejps.2013.10.018
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Development of an anti-claudin-3 and -4 bispecific monoclonal antibody for cancer diagnosis and therapy (2014)
  • Author(s): Xiangru Li, Manami Iida, Minoru Tada, Akihiro Watari, Yumi Kawahigashi, Yuka Kimura, Taku Yamashita, Akiko Ishii-Watabe, Tadayuki Uno, Masayoshi Fukasawa, Hiroki Kuniyasu, Kiyohito Yagi, Masuo Kondoh
  • Journal Title: J Pharmacol Exp Ther Volume: 351 Pages: 206-213
  • DOI: 10.1124/jpet.114.216911
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Anti-human claudin-1 monoclonal antbodies as anti-Hepatitis C Virus agents (2014)
  • Author(s): M. Iida, M. Yamashita, S. Nagase, M. Tada, Y. Shirasago, M. Fukasawa, A.Watari, A.Ishii-Watabe, K. Yagi, M. Kondoh
  • Organizer: Antibody Engineering & Therapeutics 2014
  • Place of Presentation: Huntington Beach, CA, U.S.A.
  • Year and Date: 2014-12-07 – 2014-12-11
  • Invited
• [Presentation] Epitope mapping of an anti-claudin-1 monoclonal antibody and its application to preclinical drug discovery (2014)
  • Author(s): T. Hata, S. Nagase, Y. Shirasago, M. Fukasawa, A. Watari, K. Yagi, M. Kondoh
  • Organizer: Antibody Engineering & Therapeutics 2014
  • Place of Presentation: Huntington Beach, CA, U.S.A.
  • Year and Date: 2014-12-07 – 2014-12-11
  • Invited
• [Presentation] Development of a Novel Bispecific Anti-Claudin Antibody and its Anti-Tumor Activity (2014)
  • Author(s): Y. Kimura, X. Li, M. Iida, M. Tada, A. Watabe-Ishii, M. Fukasawa, Y. Kawahigashi, A. Watari, K. Yagi, M. Kondoh
  • Organizer: Antibody Engineering & Therapeutics 2014
  • Place of Presentation: Huntington Beach, CA, U.S.A.
  • Year and Date: 2014-12-07 – 2014-12-11
  • Invited
• [Presentation] Paracellular or transcellular? Promising approach for oral delivery of macromolecules (2014)
  • Author(s): Masuo Kondoh, Kiyohito Yagi
  • Organizer: 2014 AAPS Annual Meeting and Exposition
  • Place of Presentation: SanDiego, U.S.A.
  • Year and Date: 2014-11-05
  • Invited
• [Presentation] In vivo inhibition of hepatitis C virus infection by anti-claudin-1 monoclonal antibodies (2014)
  • Author(s): M.Iida, S.Nagase, M.Yamashita, Y.Shirasago, M.Fukasawa, M.Tada, A.Ishii, A.Watari, K.Yagi, and M.Kondoh
  • Organizer: The 41st Annual Meeting & Exposition of the controlled Release Society
  • Place of Presentation: Chicago, U.S.A.
  • Year and Date: 2014-07-13 – 2014-07-16
  • Invited
• [Presentation] In vivo inhibition of hepatitis C virus infection by anti-Claudin-1 monoconal antibodies (2014)
  • Author(s): M.Iida, S.Nagase, M.Yamashita, Y.Shirasago, M.Fukasawa, M.Tada, A.Ishii, A.Watari,
  • Organizer: Experimental Biology 2014
  • Place of Presentation: SanDiego,U.S.A.
  • Year and Date: 2014-04-26 – 2014-04-30
  • Invited
• [Presentation] Modulation of the epithelial bariier by a fragment of Clostridium perfringens iota-toxin (2014)
  • Author(s): D.Iguchi, T.Hayaishi, S.M. Krug, A.Watari, M.Fromm, K.Yagi and M.Kondoh
  • Organizer: Experimental Biology 2014
  • Place of Presentation: SanDiego,U.S.A.
  • Year and Date: 2014-04-26 – 2014-04-30
  • Invited
• [Presentation] Development and anti-tumor activities of claudin-4-specific monoclonal antibodies (2014)
  • Author(s): Y.Kawahigashi, X.Li, H.Kuniyasu, M.Fukasawa, M.Tada, A.Ishii, A.Watari, K.Yagi and
  • Organizer: Experimental Biology 2014
  • Place of Presentation: SanDiego,U.S.A.
  • Year and Date: 2014-04-26 – 2014-04-30
  • Invited
• [Presentation] Tumor-targeting and anti-tumor activity of anti-claucin-3/-4 antibody (2014)
  • Author(s): Y.Kimura, X.Li, M.Iida, M.Tada, A.Ishii, A.Watari, M.Fukasawa, K.Yagi and M.Kondoh
  • Organizer: Experimental Biology 2014
  • Place of Presentation: SanDiego,U.S.A.
  • Year and Date: 2014-04-26 – 2014-04-30
  • Invited
• [Presentation] Claudin-1 as a target for modulation of epidermal barriers (2014)
  • Author(s): M.Nakajima, M.Yamashita, A.Watari, K.Yagi, M.Fukasawa, and M.Kondoh
  • Organizer: Experimental Biology 2014
  • Place of Presentation: SanDiego,U.S.A.
  • Year and Date: 2014-04-26 – 2014-04-30
  • Invited