Mechanistic Study of Allosteric Inhibition of PPARgamma Serine Phosphorylation

• Project/Area Number: 25860090
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Drug development chemistry
• Research Institution: Showa Pharmaceutical University
• Principal Investigator: ITOH Toshimasa 昭和薬科大学, 薬学部, 准教授 (80536110)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2015: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: PPARgamma / 構造生物学 / メディシナルケミストリー / 高度不飽和脂肪酸 / PPAR / X線結晶構造解析 / タンパク質NMR / インスリン抵抗性 / インスリン抵抗性改善作用 / 脂肪酸合成 / PPARγ / X-線結晶構造解析

Outline of Final Research Achievements

To understand how PPARgamma ligands inhibit Ser245 phosphorylation, we have done several experiments. 1H-15N HSQC NMR experiment showed that some residues were significantly shifted by PPARgamma ligand. A signaling pathway from ligand to the shifted residues was revealed by using PPARgamma mutants. This is supported by circular dichroism spectroscopic experiments. To integrate our results, we used modeling software Sybyl-X and we proposed the main signaling pathway and mechanism of inhibition of Ser245 phspholylation by ligand.
Based on the mechanism, we planed to design and synthesis of partial agonist for PPARgamma. We established facile synthesis of 17-(S)HDHA then synthesized 17-oxoDHA, which showed PPARgamma partial agonistic activity, as we predicted.

Research Products

• [Int'l Joint Research] Italian National Research Council(イタリア)
• [Journal Article] Gram scale synthesis of specialized pro-resolving mediator 17(S)-HDHA using lipoxygenase enhanced by water-soluble reducing agent TCEP (2016)
  • Author(s): Toshimasa Itoh, Tomoko Saito, Yoshinori Yamamoto, Hiroaki Ishida, Keiko Yamamoto
  • Journal Title: Bioorganic & Medicinal Chemistry Letters Volume: 26 Issue: 2 Pages: 343345-343345
  • DOI: 10.1016/j.bmcl.2015.12.011
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Characterization of Covalent Bond Formation between PPARγ and Oxo-Fatty Acids (2015)
  • Author(s): Daichi Egawa, Toshimasa Itoh, and Keiko Yamamoto
  • Journal Title: Bioconjugate Chemistry Volume: 26 Issue: 4 Pages: 690698-690698
  • DOI: 10.1021/acs.bioconjchem.5b00021
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] リガンドによるPPARγリン酸化阻害機構における ヘリックス３とβシート間の相互作用に関する研究 (2014)
  • Author(s): 西方貴美奈, 伊藤俊将, 江川大地, 山本恵子
  • Organizer: 第58回日本薬学会関東支部大会
  • Place of Presentation: 昭和薬科大学，東京
  • Year and Date: 2014-10-04
• [Presentation] 核内受容体PPARγセリンリン酸化におけるアロステリック阻害機構解明に関する研究 (2014)
  • Author(s): 西方貴美奈, 伊藤俊将, 江川大地, 粟飯島広乃, Roberta Montanari, Louise Fairall, John Schwabe, Fulvio Loiodice, Giorgio Pochetti, 山本恵子
  • Organizer: PFシンポジウム
  • Place of Presentation: つくば国際会議場 (つくば市）
• [Presentation] Mechanistic Study of Allosteric Inhibition of PPARγ Serine Phosphorylation (2013)
  • Author(s): Toshimasa Itoh, Hirono Awaishima, Daichi Egawa, Roberta Montanari, Fulvio Loiodice , Giorgio Pochetti, and Keiko Yamamoto
  • Organizer: 14th Tetrahedron Symposium
  • Place of Presentation: Hilton Vienna (Vienna, Austria)
• [Presentation] Toshimasa Itoh, Daichi Egawa, Hirono Awaishima, Kimina Nishikata, Roberta Montanari, Louise Fairall, John Schwabe, Fulvio Loiodice, Giorgio Pochetti, Keiko Yamamoto (2013)
  • Author(s): Mechanistic Study of Allosteric Inhibition of PPARγ Serine Phosphorylation
  • Organizer: EMBO Conference
  • Place of Presentation: Hilton Sorrento Palace (Sorrento, Italy)
• [Remarks] 総説・総合論文・著書
  • URL: http://www.shoyaku.ac.jp/labosite/iyaku/2015/03/post_3.html
• [Remarks] 原著論文 (2003-)
  • URL: http://www.shoyaku.ac.jp/labosite/iyaku/2015/03/_2003_1.html