Development of cell-penetrating peptides and their application as DDS carriers

• Project/Area Number: 25713008
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Drug development chemistry
• Research Institution: Nagasaki University
• Principal Investigator: OBA Makoto 長崎大学, 医歯薬学総合研究科(薬学系), 准教授 (20396716)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥24,050,000 (Direct Cost: ¥18,500,000、Indirect Cost: ¥5,550,000); Fiscal Year 2016: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2015: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2014: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2013: ¥12,740,000 (Direct Cost: ¥9,800,000、Indirect Cost: ¥2,940,000)
• Keywords: ペプチド / ドラッグデリバリーシステム / 非天然型アミノ酸 / コンフォメーション / 膜透過性ペプチド / 有機化学 / 薬学 / バイオテクノロジー

Outline of Final Research Achievements

I developed cell-penetrating peptides (CPPs) with alpha,alpha-disubstituted alpha-amino acids (dAAs), which are one of the unnatural amino acids. Introduction of dAAs into the peptides stabilized their secondary structures as a helix even in aqueous solution. The dAAs-containing peptides showed high cell-penetrating ability compared to conventional CPPs and delivered plsmid DNA into the cultured cells efficiently.

Research Products

• [Int'l Joint Research] IECB(フランス)
• [Journal Article] Conformational studies on peptides having dipropylglycine (Dpg) or 1-aminocycloheptanecarboxylic acid (Ac7c) within the sequence of L-leucine (Leu) residues (2016)
  • Author(s): M. Oba, H. Nonaka, M. Doi, M. Tanaka
  • Journal Title: Biopolymers (Pept.Sci.) Volume: 106 Issue: 2 Pages: 210-218
  • DOI: 10.1002/bip.22810
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Cyclic a,a-disubstituted a-amino acids with menthone in their side chains through an acetal moiety and helical structures of their peptides (2016)
  • Author(s): Kaori Furukawa, Makoto Oba, George Ouma Opiyo, Mitsunobu Doi, Masakazu Tanaka
  • Journal Title: European Journal of Organic Chemistry Volume: 16 Issue: 17 Pages: 2988-2998
  • DOI: 10.1002/ejoc.201600241
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Plasmid DNA delivery by arginine-rich cell-penetrating peptides containing unnatural amino acids (2016)
  • Author(s): T. Kato, H. Yamashita, T. Misawa, K. Nishida, M. Kurihara, M. Tanaka, Y. Demizu, M. Oba
  • Journal Title: Bioorganic & Medicinal Chemistry Volume: 24 Issue: 12 Pages: 2681-2687
  • DOI: 10.1016/j.bmc.2016.04.031
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Development of a Cell-penetrating Peptide that Exhibits Responsive Changes in its Secondary Structure in the Cellular Environment (2016)
  • Author(s): H. Yamashita, T. Kato, M. Oba, T. Misawa, T. Hattori, N. Ohoka, M. Tanaka, M. Naito, M. Kurihara, Y. Demizu
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 33003-33003
  • DOI: 10.1038/srep33003
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] A Cell-Penetrating Peptide with a Guanidinylethyl Amine Structure Directed to Gene Delivery (2016)
  • Author(s): M. Oba, T. Kato, K. Furukawa, M. Tanaka,
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 19913-19913
  • DOI: 10.1038/srep19913
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Plasmid DNA delivery using fluorescein-labeled arginine-rich peptides (2015)
  • Author(s): M. Oba,* Y. Demizu, H. Yamashita, M. Kurihara, M. Tanaka
  • Journal Title: Bioorg. Med. Chem. Volume: 23 Issue: 15 Pages: 4911-4918
  • DOI: 10.1016/j.bmc.2015.05.025
  • NAID: 120006986986
  • Peer Reviewed
• [Journal Article] Helical Peptide-Foldamers Having a Chiral Five-Membered Ring Amino Acid with Two Azido Functional Groups (2014)
  • Author(s): Makoto Oba, Hiroomi Takazaki, Naomi Kawabe, Mitsunobu Doi, Yosuke Demizu, Masaaki Kurihara, Hiromu Kawakubo, Masanobu Nagano, Hiroshi Suemune, and Masakazu Tanaka
  • Journal Title: The Journal of Organic Chemistry Volume: 79 Issue: 19 Pages: 9125-9140
  • DOI: 10.1021/jo501493x
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Cell-penetrating helical peptides having L-arginines and five-membered ring α,α-disubstituted α-amino acids. (2014)
  • Author(s): Takuma Kato, Makoto Oba, Koyo Nishida, Masakazu Tanaka
  • Journal Title: Bioconjugate Chemistry Volume: 25 Issue: 10 Pages: 1761-1768
  • DOI: 10.1021/bc5003949
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Conformational studies on peptides having chiral five-membered ring amino acid with two azido or triazole functional groups within the sequence of Aib residues (2014)
  • Author(s): Makoto Oba, Naomi Kawabe, Hiroomi Takazaki, Yosuke Demizu, Mitsunobu Doi, Masaaki Kurihara, Hiroshi Suemune, Masakazu Tanaka
  • Journal Title: Tetrahedron Volume: 70 Issue: 46 Pages: 8900-8907
  • DOI: 10.1016/j.tet.2014.09.086
  • NAID: 120006987171
  • Peer Reviewed
• [Journal Article] Amphipathic short helix-stabilized peptides with cell-membrane penetrating ability (2014)
  • Author(s): Hiroko Yamashita, Yosuke Demizu, Takuji Shoda, Yukiko Sato, Makoto Oba, Masakazu Tanaka, Masaaki Kurihara
  • Journal Title: Bioorganic & Medicinal Chemistry Volume: 22 Issue: 8 Pages: 2403-2408
  • DOI: 10.1016/j.bmc.2014.03.005
  • Peer Reviewed
• [Journal Article] Helical Oligomers with a Changeable Chiral Acetal Moiety (2013)
  • Author(s): M. Oba
  • Journal Title: Eur. J. Org. Chem. Volume: - Issue: 34 Pages: 7679-7682
  • DOI: 10.1002/ejoc.201301450
  • Peer Reviewed
• [Journal Article] Protein transfection study using multicellular tumor spheroids of human hepatoma Huh-7 cells (2013)
  • Author(s): Takuma Kato, Masakazu Tanaka, Makoto Oba
  • Journal Title: PLoS ONE Volume: 8 Issue: 12 Pages: e82876-e82876
  • DOI: 10.1371/journal.pone.0082876
  • NAID: 120006986750
  • Peer Reviewed
• [Presentation] Cell-penetrating peptide foldamers containing a,a-disubstituted a-amino acids directed to plasmid DNA delivery (2016)
  • Author(s): Makoto Oba, Takuma Kato, Masakazu Tanaka
  • Organizer: Symposium on Foldamers 2016
  • Place of Presentation: Bordeaux-Pessac, France
  • Year and Date: 2016-09-26
  • Int'l Joint Research
• [Presentation] Gene delivery using a cell-penetrating peptide with a guanidinylethyl amine structure (2016)
  • Author(s): Makoto Oba, Kaori Furukawa, Takuma Kato, Masakazu Tanaka
  • Organizer: The 8th Takeda Science Foundation Symposium on PharmaSciences, Biomolecule-Based Medicinal Science: Featuring Mid-Size Drugs
  • Place of Presentation: 武田薬品研修所（大阪府・吹田市）
  • Year and Date: 2016-01-21
  • Int'l Joint Research
• [Presentation] Cell-penetrating ability and plasmid DNA delivery using fluorescein-labeled arginine-rich peptides (2015)
  • Author(s): Makoto Oba, Kaori Furukawa, Takuma Kato, Yosuke Demizu, Hiroko Yamashita, Masaaki Kurihara, Masakazu Tanaka
  • Organizer: 11th Australian Peptide Conference
  • Place of Presentation: Kingscliff (Australia)
  • Year and Date: 2015-10-25
  • Int'l Joint Research
• [Presentation] ドラッグデリバリーペプチドの開発 (2015)
  • Author(s): 大庭 誠
  • Organizer: BioJapan 2015
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2015-10-14
• [Presentation] アルギニン含有ペプチドの細胞膜透過性とpDNAデリバリーに関する研究 (2015)
  • Author(s): 大庭 誠、古川かほり、加藤巧馬、山下博子、出水庸介、栗原正明、田中正一
  • Organizer: 第31回日本DDS学会学術集会
  • Place of Presentation: 京王プラザホテル（東京都・新宿区）
  • Year and Date: 2015-07-02
• [Presentation] グアニジニルエチルアミン構造を側鎖に有する非天然型アミノ酸を利用した膜透過性ペプチドの開発 (2015)
  • Author(s): 大庭 誠、古川かほり、加藤巧馬、田中正一
  • Organizer: 日本ケミカルバイオロジー学会第10回年会
  • Place of Presentation: 東北大学（宮城県・仙台市）
  • Year and Date: 2015-06-10
• [Presentation] 五員環状ジ置換アミノ酸を利用した新規膜透過性ペプチドの開発 (2015)
  • Author(s): 加藤巧馬、大庭 誠、西田孝洋、田中正一
  • Organizer: 日本薬学会第135年会
  • Place of Presentation: 神戸 神戸学院大学、他
  • Year and Date: 2015-03-25 – 2015-03-28
• [Presentation] 五員環状アミノ酸を含有する膜透過性ヘリカルペプチドの開発 (2014)
  • Author(s): 大庭 誠、加藤巧馬、田中正一
  • Organizer: 第32回メディシナルケミストリーシンポジウム
  • Place of Presentation: 神戸 神戸国際会議場
  • Year and Date: 2014-11-26 – 2014-11-28
• [Presentation] Evaluation of cell-penetrating ability of helical peptides having achiral five-membered α,α-disubstituted amino acids (2014)
  • Author(s): Takuma Kato, Makoto Oba, Masakazu Tanaka
  • Organizer: 33rd European Peptide Symposium
  • Place of Presentation: ブルガリア ソフィア
  • Year and Date: 2014-08-31 – 2014-09-05
• [Presentation] 五員環状アミノ酸を利用した細胞膜透過性ヘリカルペプチドの開発 (2014)
  • Author(s): 加藤巧馬、大庭 誠、田中正一
  • Organizer: 第30回日本DDS学会学術集会
  • Place of Presentation: 東京 慶応大学
  • Year and Date: 2014-07-30 – 2014-07-31
• [Presentation] アキラルな五員環状アミノ酸を含有する細胞膜透過性ヘリカルペプチドの評価 (2014)
  • Author(s): 加藤巧馬、大庭 誠、田中正一
  • Organizer: 日本薬学会第134年会
  • Place of Presentation: 熊本県熊本市熊本大学
• [Presentation] 細胞膜透過性ヘリカルペプチドの創製 (2014)
  • Author(s): 山下博子、出水庸介、佐藤由紀子、大庭 誠、田中正一、栗原正明
  • Organizer: 日本薬学会第134年会
  • Place of Presentation: 熊本県熊本市熊本大学
• [Presentation] 細胞膜透過性ヘリカルペプチドの開発 (2013)
  • Author(s): 大庭 誠、加藤巧馬、田中正一
  • Organizer: 第29回日本DDS学会学術集会
  • Place of Presentation: 京都府京都市京都テルサ
• [Presentation] 五員環状アミノ酸を含有するヘリカルペプチドの細胞膜透過性の評価 (2013)
  • Author(s): 加藤巧馬、大庭 誠、田中正一
  • Organizer: 第30回日本薬学会九州支部大会
  • Place of Presentation: 長崎県佐世保市長崎国際大学
• [Presentation] ジ置換アミノ酸を含有する細胞膜透過性ヘリカルペプチドの開発
  • Author(s): 加藤巧馬、楠富伊織、大庭 誠、田中正一
  • Organizer: 第50回化学関連支部合同九州大会
  • Place of Presentation: 福岡県北九州市北九州国際会議場
• [Remarks] 長崎大学大学院医歯薬学総合研究科生命薬科学専攻分子創薬科学講座薬化学分野
  • URL: http://www.ph.nagasaki-u.ac.jp/lab/biomimic/index-j.html
• [Remarks] 長崎大学研究者総覧データベース
  • URL: http://research.jimu.nagasaki-u.ac.jp/IST?ISTActId=FINDJPDetail&ISTKidoKbn=&ISTErrorChkKbn=&ISTFormSetKbn=&ISTTokenChkKbn=&userId=100000568
• [Patent(Industrial Property Rights)] 新規膜透過性ペプチド (2015)
  • Inventor(s): 大庭 誠、田中正一
  • Industrial Property Rights Holder: 大庭 誠、田中正一
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2015-072030
  • Filing Date: 2015-03-31