Development of cell-permeable oligonucleotides bearing bio-cleavable protecting groups

• Project/Area Number: 17K01966
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Chemical biology
• Research Institution: Shiga University of Medical Science (2019); Kanagawa University (2017-2018)
• Principal Investigator: Hisao Saneyoshi 滋賀医科大学, 医学部, 助教 (10564784)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2017: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 保護基 / 細胞膜透過性 / 細胞内化学反応 / 活性化 / 合成核酸 / プロドラッグ / 徐放性 / 発蛍光色素 / 細胞内導入 / 核酸医薬 / グルタチオン / 核酸合成化学 / 細胞内導入法 / 細胞内環境

Outline of Final Research Achievements

Objective　of　this　project　is　to　develop　synthetic　oligonucleotides　with higｈ　cell-membrane　permeability and activatable property. In research period, glutathione-labile protecting groups for phosphodiester moiety in oligonucleotides were developed. Oligonucleotides modified with glutathione-labile protecting groups showed cell-membrane permeability without use of transfection reagent. The cell-membrane permeability increased depending on the number of protecting groups. Moreover the location of protecting groups was also influenced on the cellular uptake. Internalized oligonucleoides were activated and transformed into biologically active oligonucleotides.

Academic Significance and Societal Importance of the Research Achievements

本研究成果の学術的意義は、チオールによって速やかに除去されるリン酸ジエステル部位保護基を開発した点である。保護基を結合した合成核酸は、細胞膜透過性を示しただけではなく細胞内部での脱保護反応により活性化され機能を示した。これらの成果は、核酸を基盤構造とする細胞内部で機能するプローブ分子や医薬品などへの応用が期待される。

Research Products

• [Journal Article] Thiol-responsive pro-fluorophore labeling: Synthesis of a pro-fluorescent labeled oligonucleotide for monitoring cellular uptake (2020)
  • Author(s): Saneyoshi Hisao、Yamamoto Yuta、Ohta Takayuki、Akai Shoji、Ono Akira
  • Journal Title: Bioorganic & Medicinal Chemistry Letters Volume: 30 Issue: 13 Pages: 127222-127222
  • DOI: 10.1016/j.bmcl.2020.127222
  • Peer Reviewed
• [Journal Article] 1.Design, Synthesis and Cellular Uptake of Oligonucleotides Bearing Glutathione-Labile Protecting Groups (2019)
  • Author(s): Hisao Saneyoshi, Takayuki Ohta, Yuki Hiyoshi, Takeo Saneyoshi, Akira Ono.
  • Journal Title: Organic Letters Volume: 21 Issue: 4 Pages: 862-866
  • DOI: 10.1021/acs.orglett.8b03501
  • Peer Reviewed / Open Access
• [Journal Article] Prodrug型核酸医薬に用いる保護基の開発 (2019)
  • Author(s): 實吉尚郎、小野晶
  • Journal Title: Medical Science Digest Volume: 45 Pages: 4-7
• [Journal Article] Development of Protecting Groups for Prodrug-Type Oligonucleotide Medicines (2018)
  • Author(s): Hisao Saneyoshi, Akira Ono
  • Journal Title: Chemical and Pharmaceutical Bulletin Volume: 66 Issue: 2 Pages: 147-154
  • DOI: 10.1248/cpb.c17-00696
  • NAID: 130006328065
  • ISSN: 0009-2363, 1347-5223
  • Peer Reviewed / Open Access
• [Presentation] 細胞内で除去される保護基を結合したプロドラッグ型核酸医薬の合成手法の開発研究 (2020)
  • Author(s): 小野 晶、三上 智紀、鈴木 海斗、堀川 匡嗣、寺澤 一馬、太田 貴之、中村 康大、實吉 尚郎
  • Organizer: 日本薬学会第140年会
• [Presentation] 細胞内で活性化されて機能するオリゴヌクレオチドの創製 (2019)
  • Author(s): 實吉尚郎
  • Organizer: 第17回有機合成化学協会関西支部賞受賞講演会
  • Invited
• [Presentation] Development of a reduction cleavable linker for solid-phase synthesis (2019)
  • Author(s): Kazuma Terasawa, Akira Ono, Hisao Saneyoshi
  • Organizer: 第46回国際核酸化学シンポジウム
  • Int'l Joint Research
• [Presentation] Synthesis of cell-permeable and GSH-activatable oligonucleotides (2019)
  • Author(s): Takayuki Ohta, Zhaoma Shu, Hiroshi Abe, Akira Ono, Hisao Saneyoshi
  • Organizer: 第46回国際核酸化学シンポジウム
  • Int'l Joint Research
• [Presentation] 細胞内還元条件で除去されるRNA糖部保護基の開発 (2019)
  • Author(s): 寺澤一馬、中村康大、小野晶、實吉尚郎
  • Organizer: 日本核酸医薬学会第5回年会
• [Presentation] 細胞内で蛍光を発する発光標識核酸の合成 (2019)
  • Author(s): 太田貴之、山本祐太、小野晶、實吉尚郎
  • Organizer: 日本核酸医薬学会第5回年会
• [Presentation] 発蛍光色素が結合したオリゴヌクレオチドの合成 (2019)
  • Author(s): 實吉尚郎、山本祐太、太田貴之、小野晶
  • Organizer: 日本化学会第99春季年会
• [Presentation] 2' 位に還元環境で脱離する保護基を結合した RNA の合成：プロドラッグ型核酸医薬開発に向けて (2019)
  • Author(s): 小野晶、中村康大、實吉尚郎
  • Organizer: 日本薬学会第139年会
• [Presentation] Synthesis of cell-membrane permeable oligonucleotides bearing GSH-activated protecting groups on the internucleotide linkages (2018)
  • Author(s): Hisao Saneyoshi, Takayuki Ohta, Yuki Hiyoshi, Akira Ono
  • Organizer: 第45回国際核酸化学シンポジウム
  • Int'l Joint Research
• [Presentation] 實吉尚郎、太田貴之、日吉祐貴、小野晶 (2018)
  • Author(s): プロドラッグ型核酸の合成と細胞内取り込み
  • Organizer: 第12回バイオ関連化学シンポジウム
• [Presentation] Development of reduction-activated protecting groups for siRNA prodrugs (2018)
  • Author(s): Kodai Nakamura, Akira Ono, Hisao Saneyoshi
  • Organizer: 23th International Round Table on Nucleosides, Nucleotides and Nucleic Acids (XXIII IRT)
  • Int'l Joint Research
• [Presentation] Development of cell-permeable oligonucleotides bearing GSH-activated protecting groups (2018)
  • Author(s): Takayuki Ohta, Akira Ono, Hisao Saneyoshi
  • Organizer: 23th International Round Table on Nucleosides, Nucleotides and Nucleic Acids (XXIII IRT)
  • Int'l Joint Research
• [Presentation] Synthetic study of bio-reduction cleavable linker for oligonucleotides (2018)
  • Author(s): Kazuma Terasawa, Akira Ono, Hisao Saneyoshi
  • Organizer: 23th International Round Table on Nucleosides, Nucleotides and Nucleic Acids (XXIII IRT)
  • Int'l Joint Research
• [Presentation] 蛍光発生型色素が結合したオリゴヌクレオチドの合成 (2018)
  • Author(s): 實吉尚郎、山本祐太、太田貴之、小野晶
  • Organizer: 日本核酸医薬学会第4回年会
• [Presentation] 癌細胞内で開裂するリンカーの開発研究 (2018)
  • Author(s): 寺澤一馬、小野晶、實吉尚郎
  • Organizer: 日本核酸医薬学会第4回年会
• [Presentation] 細胞内還元条件で除去されるRNA糖部保護基の開発 (2018)
  • Author(s): 中村康大、小野晶、實吉尚郎
  • Organizer: 日本核酸医薬学会第4回年会
• [Presentation] 細胞膜透過性を有するグルタチオン応答性プロドラッグ型核酸の合成 (2018)
  • Author(s): 太田貴之、小野晶、實吉尚郎
  • Organizer: 日本核酸医薬学会第4回年会
• [Presentation] Design and synthesis of bio-labile protecting groups for oligonucleotide prodrugs (2018)
  • Author(s): Hisao Saneyoshi, Takayuki Ohta, Kazuma Terasawa, Yuta Yamamoto, Akira Ono.
  • Organizer: 第44回国際核酸化学シンポジウム
  • Int'l Joint Research
• [Presentation] Glutathione-labile protecting groups for phosphodiester moieties (2018)
  • Author(s): Takayuki Ohta, Yuta Yamamoto, Akira Ono, Hisao Saneyoshi
  • Organizer: 第44回国際核酸化学シンポジウム
  • Int'l Joint Research
• [Presentation] Synthesis of cell-permeable fluorogenic oligonucleotides (2018)
  • Author(s): Yuta Yamamoto, Akira Ono, Hisao Saneyoshi
  • Organizer: 第44回国際核酸化学シンポジウム
  • Int'l Joint Research
• [Patent(Industrial Property Rights)] 2.ヌクレオシド又はそのヌクレオチドの誘導体、それを構成単位として含むRNA誘導体、核酸医薬、及びRNA誘導体若しくはRNAの製造方法. (2018)
  • Inventor(s): 小野晶、實吉尚郎
  • Industrial Property Rights Holder: 神奈川大学
  • Industrial Property Rights Type: 特許
  • Patent Publication Number: 2018-184351
  • Filing Date: 2018
• [Patent(Industrial Property Rights)] 化合物、蛍光標識導入剤、及び蛍光標識導入方法 (2018)
  • Inventor(s): 2018
  • Industrial Property Rights Holder: 2018
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2018-089292
  • Filing Date: 2018