Synthesis of double labeled ceramide fluorescent probes to analyze metabolism of sphingolipids

• Project/Area Number: 25670003
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Chemical pharmacy
• Research Institution: Chiba University
• Principal Investigator: Nishida Atsushi 千葉大学, 薬学研究科(研究院), 教授 (80130029)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: セラミド / 代謝 / 蛍光標識 / NBD / BODIPY / ゴルジ / 脂質代謝 / 分子プローブ / ゴルジマーカー / がん / アポトーシス / スフィンゴシン / 蛍光プローブ / 二重標識

Outline of Final Research Achievements

Doubly labeled fluorescent probe at main and side chain on ceramide has been developed to obtain information of ceramide metabolism in cell. Such information will be useful to evaluate role of ceramide in connection with many diseases such as cancers. We developed first several green fluorescent ceramide probes. One of the ceramide probe which has NBD at main chain and an acety group on a primary hydroxy group was less toxic compared to commercially available C6-NBD ceramide and stain golgi system for a long period without damage on cell. We also developed a red fluorescent ceramide probe which has a BODIPY type group at main chain of ceramide. We also observed energy transfer between two florescent group such as NBD and BODIPY in the same ceramide molecule. Those results will be useful to develop a new fluorescent probes which is useful to analyze ceramide metabolism in living cells.

Research Products

• [Journal Article] Trafficking of acetyl-C16-ceramide-NBD with long-term stability and no cytotoxicity into the Golgi. (2015)
  • Author(s): Makiyama T, Nakamura H, Nagasaka N, Yamashita H, Honda T, Yamaguchi N, Nishida A, Murayama T.
  • Journal Title: Traffic Volume: 16 Issue: 5 Pages: 476-492
  • DOI: 10.1111/tra.12265
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] 長時間安定的にゴルジ体を標識する新規蛍光標識セラミド誘導体 (2016)
  • Author(s): 中村浩之、牧山智彦、長坂伸夫、山下尚大、本田拓也、山口直人、西田篤司、村 山俊彦
  • Organizer: 日本薬学会第１３６年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-03-26
• [Presentation] 新規赤色蛍光標識セラミドプローブの合成研究 (2016)
  • Author(s): 王 亜賓、楊井理信、橋本直宏、 中村浩之、村山俊彦、西田 篤司
  • Organizer: 日本薬学会第１３６年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-03-26
• [Presentation] Development of Regio-and Stereoselective Hydrocyanation of Allene and Its Application to Hydrocyanative Cyclization under Nickel Catalysis (2014)
  • Author(s): Amako,Y., Arai,S., Nishida,A
  • Organizer: The 4th Junior International Conference on Cutting-Edge Organic Chemistry in Asia
  • Place of Presentation: Bangkok, Thailand
  • Year and Date: 2014-11-28 – 2014-11-30
  • Invited
• [Presentation] Enantioselective construction of multi-substituted hydrocarbazoles by chiral holmium/bisthiourea catalyst (2014)
  • Author(s): Morikawa, T., Harada, S., Nishida, A.
  • Organizer: 15th Tetrahedron Symposium
  • Place of Presentation: Singapore
  • Year and Date: 2014-11-03 – 2014-11-05
• [Presentation] 標識セラミドの合成研究
  • Author(s): 楊井理信、長坂伸夫、牧山智彦、中村浩之、村山俊彦、西田篤司
  • Organizer: 第66回有機合成化学協会関東支部シンポジウム
  • Place of Presentation: 東京
• [Remarks] 千葉大学大学院薬学研究院薬品合成化学研究室
  • URL: http://www.p.chiba-u.jp/lab/gousei/index.html
• [Patent(Industrial Property Rights)] Ceramide Derivative and Goligi Apparatus-Labelling Fluorescent Probe Using Same (2014)
  • Inventor(s): A Nishida, et al.
  • Industrial Property Rights Holder: A Nishida, et al.
  • Industrial Property Rights Type: 特許
  • Filing Date: 2014-06-13
  • Acquisition Date: 2014-11-06
  • Overseas