バイオ医薬品の非侵襲性投与を可能とする次世代型TJ binderの開発

• Project/Area Number: 16K08370
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Medical pharmacy
• Research Institution: Osaka University
• Principal Investigator: 渡利 彰浩 大阪大学, 薬学研究科, 講師 (80452465)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Discontinued (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 吸収促進 / Tight junction / claudin / バイオ医薬品 / スクリーニング / TR-FRET / 薬学 / 医療・福祉 / 生体分子 / 細胞・組織

Outline of Annual Research Achievements

上皮細胞層は生体内外を隔てるバリアとして機能しており、恒常性を維持するなどの重要な役割を担っている。このバリア機能は細胞間結合の一つであるTight Junction（TJ）が関わっており、TJにより細胞間隙での物質透過が制御されている。一方、TJによる透過制限のため、細胞間隙での薬物吸収は困難となっている。TJでのバリア機能は主に膜タンパク質claudinが担っており、claudinを制御することによる薬物吸収促進法の開発が展開されている。本研究では、これまでに見出したウエルシュ菌下痢毒素の受容体結合領域（C-CPE）によるバイオ医薬品をも経粘膜吸収促進可能な上皮バリア制御活性を、実用性に優れた低分子化合物により実現するため、独自に構築したclaudin-4 binder検出系を利用し、高い上皮バリア機能制御活性を示す低分子claudin-4 binderの取得を目指す。
昨年度に取得した低分子claudin-4 binderのTJバリア制御活性に関して、腸管上皮モデルCaco-2細胞を用い、経上皮抵抗値（TEER）を指標としたバリア制御活性の評価を行った結果、それぞれの化合物単独ではTEERの変動は低かったが、特定の3化合物を組み合わせて評価したところC-CPEに匹敵する高いTEER低下作用が確認された。また、TEER低下時での細胞間隙透過試験を実施したところ、モデル薬物（分子量約4kDa）の透過性亢進作用が確認された。以上の結果より、複数の組み合わせでTJバリア機能制御活性を示す低分子化合物の取得に成功した。

Research Products

• [Journal Article] Pancreatic adenocarcinoma upregulated factor has oncogenic functions in oral squamous cell carcinoma (2018)
  • Author(s): Sasahira T, Kurihara M, Nishiguchi Y, Nakashima C, Kirita T, Kuniyasu H.
  • Journal Title: Histopathol Volume: 70 Pages: 539-548
  • DOI: 10.1016/j.ejps.2018.02.016
  • Peer Reviewed
• [Journal Article] Lignosulfonic acid attenuates NF-κB activation and intestinal epithelial barrier dysfunction induced by TNF-α/IFN-γ in Caco-2 cells (2017)
  • Author(s): Matsuhisa Koji、Watari Akihiro、Iwamoto Kazuki、Kondoh Masuo、Yagi Kiyohito
  • Journal Title: J Nat Med Volume: 72 Issue: 2 Pages: 448-455
  • DOI: 10.1007/s11418-017-1167-5
  • NAID: 40021511333
  • Peer Reviewed
• [Journal Article] Identification of claudin-4 binder that attenuates tight junction barrier function by TR-FRET-based screening assay (2017)
  • Author(s): Watari Akihiro、Kodaka Miki、Matsuhisa Koji、Sakamoto Yuta、Hisaie Kota、Kawashita Norihito、Takagi Tatsuya、Yamagishi Yoshiaki、Suzuki Hidehiko、Tsujino Hirofumi、Yagi Kiyohito、Kondoh Masuo
  • Journal Title: Sci Rep Volume: 7 Issue: 1 Pages: 14514-14514
  • DOI: 10.1038/s41598-017-15108-y
  • Peer Reviewed / Open Access
• [Journal Article] Dodecylmaltoside Modulates Bicellular Tight Junction Contacts To Promote Enhanced Permeability (2017)
  • Author(s): Gradauer K.、Iida M.、Watari A.、Kataoka M.、Yamashita S.、Kondoh M.、Buckley S. T.
  • Journal Title: Mol Pharm Volume: 14 Issue: 12 Pages: 4734-4740
  • DOI: 10.1021/acs.molpharmaceut.7b00297
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Creation of a Claudin-2 Binder and Its Tight Junction?Modulating Activity in a Human Intestinal Model (2017)
  • Author(s): Takigawa Mutsumi、Iida Manami、Nagase Shotaro、Suzuki Hidehiko、Watari Akihiro、Tada Minoru、Okada Yoshiaki、Doi Takefumi、Fukasawa Masayoshi、Yagi Kiyohito、Kunisawa Jun、Kondoh Masuo
  • Journal Title: J Pharmacol Exp Ther. Volume: 363 Issue: 3 Pages: 444-451
  • DOI: 10.1124/jpet.117.242214
  • Peer Reviewed
• [Journal Article] Claudin-5-Binders Enhance Permeation of Solutes across the Blood-Brain Barrier in a Mammalian Model (2017)
  • Author(s): Hashimoto Yosuke、Shirakura Keisuke、Okada Yoshiaki、Takeda Hiroyuki、Endo Kohki、Tamura Maki、Watari Akihiro、Sadamura Yoshifusa、Sawasaki Tatsuya、Doi Takefumi、Yagi Kiyohito、Kondoh Masuo
  • Journal Title: J Pharmacol Exp Ther. Volume: 363 Issue: 2 Pages: 275-283
  • DOI: 10.1124/jpet.117.243014
  • Peer Reviewed
• [Journal Article] Angubindin-1, a novel paracellular absorption enhancer acting at the tricellular tight junction. (2017)
  • Author(s): Krug SM, Hayaishi T, Iguchi D, Watari A,Takahashi A, Fromm M, Nagahama M, Takeda H, Okada Y,Sawasaki T, Doi T, Yagi K, Kondoh M
  • Journal Title: J. Control. Release. Volume: 260 Pages: 1-11
  • DOI: 10.1016/j.jconrel.2017.05.024
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Rebeccamycin attenuates TNF-α-induced intestinal epithelial barrier dysfunction by inhibiting myosin light chain kinase production (2017)
  • Author(s): Watari A., Sakamoto Y., Hisaie K., Iwamoto K., Fueta M., Yagi K., Kondoh M.
  • Journal Title: Cellular Physiology and Biochemistry Volume: -
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Efficacy and safety evaluation of claudin-4-targeted antitumor therapy using a human and mouse cross-reactive monoclonal antibody (2016)
  • Author(s): Hashimoto Y., Kawahigashi Y., Hata T., Li X., Watari A., Tada M., Ishii-Watabe A., Okada Y., Doi T., Fukasawa M., Kuniyasu H., Yagi K., Kondoh M
  • Journal Title: Pharmacology Research & Perspectives Volume: 4(5)
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Generation and characterization of a human-mouse chimeric antibody against the extracellular domain of claudin-1 for cancer therapy using a mouse model. (2016)
  • Author(s): Hashimoto Y, Tada M, Iida M, Nagase S, Hata T, Watari A, Okada Y, Doi T, Fukasawa M, Yagi K, Kondoh M.
  • Journal Title: Biochem Biophys Res Commun. Volume: 477(1) Issue: 1 Pages: 91-95
  • DOI: 10.1016/j.bbrc.2016.06.025
  • Peer Reviewed
• [Presentation] TR-FRETによる低分子 CLDN-4 binder スクリーニング系の構築 (2017)
  • Author(s): 坂本雄太、渡利彰浩、小高美樹、久家広大、八木清仁、近藤昌夫
  • Organizer: 日本薬学会 第137年会
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-25
• [Presentation] DNA damageが誘導する腸管上皮バリアの機能亢進 (2017)
  • Author(s): 渡利彰浩、長谷川真希、八木清仁、近藤昌夫
  • Organizer: 日本薬学会 第137年会
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-25
• [Presentation] Homoharringtonineによる核酸医薬の経皮投与基盤技術の開発 (2017)
  • Author(s): 藤原果奈、渡利彰浩、永田哲也、近藤昌夫、横田隆徳、八木清仁
  • Organizer: 日本薬学会 第137年会
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-25
• [Presentation] Use of a Time-Resolved Fluorescence Resonance Energy Transfer-based screening assay to identify a claudin-4 binder that attenuates tight junction barrier function. (2017)
  • Author(s): A. Watari, K. Yagi, M. Kondoh
  • Organizer: ASCB/EMBO 2017 meeting
  • Int'l Joint Research
• [Presentation] Homoharringtonine enhances transdermal absorption in the skin by regulating epidermal tight junction barriers. (2017)
  • Author(s): K. Yagi, A. Watari, K. Fujiwara, N. Suzuki, M. Kondoh
  • Organizer: ASCB/EMBO 2017 meeting
  • Int'l Joint Research
• [Presentation] Time-Resolved Fluorescence Resonance Energy Transfer-based Screen for Identification of Novel Claudin-4 binders that Modulate the Epithelial Tight Junction Barrier (2017)
  • Author(s): A. Watari, Y. Sakamoto, K. Hisaie, K. Yagi, M. Kondoh
  • Organizer: The 2017 AAPS Annual Meeting and Exposition programming
  • Int'l Joint Research
• [Presentation] Development of Homoharringtonine as a Penetration Enhancer for Transdermal Drug Administration (2017)
  • Author(s): K. Yagi, A. Watari, k. Fujiwara, T. kurachi, N. Suzuki, M. Kondoh
  • Organizer: The 2017 AAPS Annual Meeting and Exposition programming
  • Int'l Joint Research
• [Presentation] Use of a Time-Resolved Fluorescence Resonance Energy Transfer-based screen to identify a novel claudin-4 binding compound that attenuates the tight junction barrier (2017)
  • Author(s): A. Watari, K. Yagi, M. Kondoh
  • Organizer: 6th FIP Pharmaceutical Sciences World Congress 2017
  • Int'l Joint Research
• [Presentation] Time-resolved fluorescence resonance energy transfer法により同定した低分子claudin-4 binderの機能解析 (2017)
  • Author(s): 坂本雄太、渡利彰浩、小高美樹、久家広大、八木清仁、近藤昌夫
  • Organizer: 第40回日本分子生物学会
• [Presentation] タイトジャンクション構成因子を標的とした創薬基盤研究 (2017)
  • Author(s): 渡利彰浩、八木清仁、近藤昌夫
  • Organizer: 阪大薬―医薬基盤健栄研ランチミーティング
  • Invited
• [Presentation] Homoharringtonineによる経皮吸収促進効果の検討 (2017)
  • Author(s): 渡利彰浩、藤原果奈、倉智彩夢、近藤昌夫、八木清仁
  • Organizer: 第33回日本DDS学会学術集会
• [Book] Tight junction modulatorを用いたペプチド医薬品の経粘膜吸収促進法 (2017)
  • Author(s): 渡利彰浩, 八木清仁, 近藤昌夫
  • Total Pages: 9
  • Publisher: 技術情報協会
• [Book] タイトジャンクションバリア制御によるバイオ医薬品の吸収促進技術開発に向けて (2017)
  • Author(s): 渡利彰浩
  • Total Pages: 2
  • Publisher: Drug Delivery System
• [Patent(Industrial Property Rights)] 核酸を含む経腸投与用組成物 (2016)
  • Inventor(s): 八木清仁、近藤昌夫、渡利彰浩 等
  • Industrial Property Rights Holder: 八木清仁、近藤昌夫、渡利彰浩 等
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-11-30
  • Overseas