Development of macrocylclic peptide drug that targets membrane proteins using cultivated human cell lines

• Project/Area Number: 24681047
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Chemical biology
• Research Institution: The University of Tokyo
• Principal Investigator: KATOH Takayuki 東京大学, 理学(系)研究科(研究院), 助教 (90567760)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥26,910,000 (Direct Cost: ¥20,700,000、Indirect Cost: ¥6,210,000); Fiscal Year 2014: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000); Fiscal Year 2013: ¥8,580,000 (Direct Cost: ¥6,600,000、Indirect Cost: ¥1,980,000); Fiscal Year 2012: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000)
• Keywords: 膜タンパク質 / ペプチド医薬 / RaPIDディスプレイ / 翻訳 / 遺伝暗号リプログラミング / アゴニスト / 特殊環状ペプチド / 阻害剤 / RaPID display / 医薬品探索 / 分子標的医薬 / 特殊ペプチド / GPCR

Outline of Final Research Achievements

In this research project, we developed a new method for screening non-standard macrocyclic peptide drugs that target cell-surface molecules such as membrane proteins. Normally, target molecules need to be immobilized on magnetic beads for applying in-vitro display method combined with random peptide library. However, immobilization of membrane proteins is generally difficult because isolation and solubilization of such proteins are impossible in many cases. Therefore, in this project, we developed a new in-vitro display method that utilizes cultured cells and/or baculovirus expressing membrane proteins on their surface instead of purified proteins immobilized on beads. By using this method, we could successfully obtain macrocyclic peptides that target CD20, IL28RA, Claudin-1 and 4.

Research Products

• [Journal Article] Selection-Based Discovery of Druglike Macrocyclic Peptides. (2014)
  • Author(s): Passioura T, Katoh T, Goto Y, Suga H.
  • Journal Title: Annu. Rev. Biochem. Volume: 未定
  • Peer Reviewed
• [Journal Article] A macrocyclic peptide that serves as a cocrystallization ligand and inhibits the function of a MATE family transporte (2013)
  • Author(s): C・J・ Hipolito, Y・Tanaka, T・ Katoh, O・ Nureki, H・ Suga
  • Journal Title: Molecules Volume: 18 Issue: 9 Pages: 10514-30
  • DOI: 10.3390/molecules180910514
  • Peer Reviewed
• [Journal Article] Structural basis for the drug extrusion mechanism by a MATE multidrug transporter (2013)
  • Author(s): Y・ Tanaka, C・J・ Hipolito, A・D・ Maturana, K・ Ito, T・ Kuroda, T・ Higuchi, T・ Katoh, H・E・ Kato, M・ Hattori, K・ Kumazaki, T・ Tsukazaki, R・ Ishitani, H・ Suga, O・ Nureki
  • Journal Title: Nature Volume: 496 Issue: 7444 Pages: 247-51
  • DOI: 10.1038/nature12014
  • Peer Reviewed
• [Journal Article] RNAワールドの実験的検証 (2013)
  • Author(s): 岩根由彦、高辻諒、加藤敬行、菅裕明
  • Journal Title: 進化分子工学 Volume: なし Pages: 49-60
• [Journal Article] Selective thioether macrocyclization of peptides having the N-terminal 2-chloroacetyl group and competing two or three cysteine residues in translation (2012)
  • Author(s): Kazuhiro Iwasaki, Yuki Goto, Takayuki Katoh, Hiroaki Suga
  • Journal Title: Org.Biomol.Chem. Volume: 10 Issue: 30 Pages: 5783-5786
  • DOI: 10.1039/c2ob25306b
  • Peer Reviewed
• [Presentation] 細胞を用いたRaPIDシステムによるIL28RA結合ペプチドの探索 (2014)
  • Author(s): 石田啓、平田雄一、加藤敬行、小原道法、菅裕明
  • Organizer: 第46回 若手ペプチド夏の勉強会
  • Place of Presentation: 京都府立青少年海洋センター（京都府宮津市）
  • Year and Date: 2014-08-03 – 2014-08-05
• [Presentation] 特殊環状ペプチド創薬 (2013)
  • Author(s): 加藤敬行
  • Organizer: 日本生化学会
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Invited
• [Book] 生命システム工学 (2012)
  • Author(s): 加藤敬行、後藤佑樹、菅裕明, 他
  • Total Pages: 213
  • Publisher: 化学同人