Drug development of African sleeping sickness targeting energy metabolic pathway

• Project/Area Number: 26440027
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Kyoto Institute of Technology
• Principal Investigator: Shiba Tomoo 京都工芸繊維大学, 応用生物学系, 准教授 (80401206)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: トリパノソーマ / 顧みられない熱帯病 / エネルギー代謝 / 二核鉄タンパク質 / 膜表剤型タンパク質 / アスコフラノン / X線結晶構造解析 / 顧みられい熱帯病 / diironタンパク質 / 膜表在型タンパク質 / Ｘ線結晶構造解析

Outline of Final Research Achievements

The trypanosome cyanide-insensitive alternative oxidase (TAO) and trypanosome glycerol kinase (TGK) are drug targets for human African sleeping sickness. In this study, I determined the crystal structures of TAO and TGK in complex with its lead compounds to make more potent drug. The crystal structure of TAO in complex with its new inhibitor, ferulenol was determined at 2.7 Å resolution. I found GK-17b which is a novel inhibitor of TGK. In addition, this compound inhibits TAO enzymatic activity with comparable strength.

Research Products

• [Journal Article] Conjugates of 2,4-Dihydroxybenzoate and Salicylhydroxamate and Lipocations Display Potent Antiparasite Effects by Efficiently Targeting the Trypanosoma brucei and Trypanosoma congolense Mitochondrion. (2017)
  • Author(s): Francisco Jose Fueyo Gonzalez, Godwin U. Ebiloma, Carolina Izquierdo Garcia, Victor Bruggeman, Jose Maria Sanchez Villamanan, Anne Donachie, Emmanuel Oluwadare Balogun, Daniel Ken Inaoka, Tomoo Shiba, Shigeharu Harada, Kiyoshi Kita, Harry P. de Koning, Christophe Dardonville
  • Journal Title: Journal of Medicinal Chemistry Volume: 60(4) Issue: 4 Pages: 1509-1522
  • DOI: 10.1021/acs.jmedchem.6b01740
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The hyperthermophilic cystathionine γ-synthase from the aerobic crenarchaeon Sulfolobus tokodaii: expression, purification, crystallization and structural insights (2017)
  • Author(s): Sato D, Shiba T, Mizuno S, Kawamura A, Hanada S, Yamada T, Shinozaki M, Yanagitani M, Tamura T, Inagaki K, Harada S
  • Journal Title: Acta Cryst., F Volume: 73 Issue: 3 Pages: 152-158
  • DOI: 10.1107/s2053230x17002011
  • Peer Reviewed / Open Access
• [Journal Article] Structural and mechanistic insights into homocysteine degradation by a mutant of methionine γ-lyase based on substrate-assisted catalysis (2017)
  • Author(s): Sato D, Shiba T, Yunoto S, Furutani K, Fukumoto M, Kudou D, Tamura T, Inagaki K, Harada S.
  • Journal Title: Protein Science Volume: 26 Issue: 6 Pages: 152-158
  • DOI: 10.1002/pro.3158
  • NAID: 130007628790
  • Peer Reviewed / Open Access
• [Journal Article] 1.Structural Insights into the Molecular Design of Flutolanil Derivatives Targeted for Fumarate Respiration of Parasite Mitochondria (2015)
  • Author(s): Inaoka DK, Shiba T, Sato D, Balogun EO, Sasaki T, Nagahama M, Oda M, Matsuoka S, Ohmori J, Honma T, Inoue M, Kita K, Harada S
  • Journal Title: Int. J. Mol. Sci. Volume: 16 Issue: 7 Pages: 15287-15308
  • DOI: 10.3390/ijms160715287
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] 2.Pharmacophore Modeling for Anti-Chagas Drug Design Using the Fragment Molecular Orbital Method (2015)
  • Author(s): Yoshino R, Yasuo N, Inaoka DK, Hagiwara Y, Ohno K, Orita M, Inoue M, Shiba T, Harada S, Honma T, Balogun EO, da Rocha JR, Montanari CA, Kita K, Sekijima M
  • Journal Title: PLoS One Volume: 10 Issue: 5 Pages: 1-15
  • DOI: 10.1371/journal.pone.0125829
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Probing the ubiquinol-binding site of recombinant Sauromatum guttatum alternative oxidase expressed in E. coli membranes through site-directed mutagenesis (2014)
  • Author(s): Luke Young, Benjamin May, Alice Pendlebury-Watt, Julia Shearman, Catherine Elliott, Mary S. Albury, Tomoo Shiba, Daniel Ken Inaoka, Shigeharu Harada, Kiyoshi Kita, Anthony L. Moore
  • Journal Title: Biochimica et Biophysica Acta (BBA) - Bioenergetics Volume: 1837 Issue: 7 Pages: 1219-1225
  • DOI: 10.1016/j.bbabio.2014.01.027
  • Peer Reviewed
• [Journal Article] Molecular basis for the reverse reaction of African human trypanosomes glycerol kinase (2014)
  • Author(s): Emmanuel Oluwadare Balogun, Daniel Ken Inaoka, Tomoo Shiba, Yasutoshi Kido, Chiaki Tsuge, Takeshi Nara, Takashi Aoki, Teruki Honma, Akiko Tanaka, Masayuki Inoue, Shigeru Matsuoka, Paul A.M. Michels, Kiyoshi Kita, Shigeharu Harada
  • Journal Title: Molecular Microbiology Volume: 94 Issue: 6 Pages: 1315-1329
  • DOI: 10.1111/mmi.12831
  • Peer Reviewed
• [Journal Article] 微生物からの贈り物アスコフラノン - アフリカ睡眠病からがん細胞まで - (2014)
  • Author(s): 北潔, 志波智生, 斎本博之, 稲岡ダニエル健, 原田繁春
  • Journal Title: MEDCHEM NEWS Volume: 24-4 Pages: 44-50
  • NAID: 130007674627
  • Peer Reviewed
• [Presentation] Serendipitous discovery of trypanocidal compounds with dual-inhibition for African trypanosomes glycerol kinase and alternative oxidase (2016)
  • Author(s): Emmnuel O. Balogun, Daniel K. Inaoka, Tomoo Shiba, Chiaki Tsuge, Benjamin May, Tomohiro Sato, Yasutoshi Kido, Takeshi Nara, Takashi Aoki, Teruki Honma, Akiko Tanaka, Masayuki Inoue, Shigeru Matsuoka, Paul A.M. Michels, Anthony L Moore, Shigeharu Harada, Kiyoshi Kita
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台
  • Year and Date: 2016-09-25
• [Presentation] アフリカトリパノソーマのシアン耐性酸化酵素(TAO)とクマリン骨格をもつ新規阻害剤との複合体結晶構造 (2016)
  • Author(s): 西藤万智, 志波智生, 上田慧, 稲岡ダニエル健, 城戸康年, バログン エマニエル, アントニー ムーア, 斎本博之, 北潔, 原田繁春
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台
  • Year and Date: 2016-09-25
• [Presentation] トリパノソーマシアン耐性酸化酵素(TAO)とグリセロールキナーゼ(GK)の結晶構造 (2015)
  • Author(s): 志波智生、エマニュエルバログン、稲岡ダニエル健、原田繁春、北潔
  • Organizer: 第41回生体エネルギー研究会討論会
  • Place of Presentation: 東京
  • Year and Date: 2015-12-21
• [Presentation] The crystal structures of trypanosomal cyanide-insensitive alternative oxidase (TAO) and glycerol kinase (GK) (2015)
  • Author(s): Tomoo Shiba, Emmanuel Oluwadare Balogun, Daniel Ken Inaoka, Shigeharu Harada, Kiyoshi Kita
  • Organizer: 3rd International Picobiology Institute Symposium
  • Place of Presentation: 兵庫県立大学
  • Year and Date: 2015-12-08
  • Int'l Joint Research
• [Presentation] アフリカトリパノソーマのシアン耐性酸化酵素(TAO)と異なる骨格を持つ阻害剤、アスコフラノン及びフェルレノールとの複合体構造 (2015)
  • Author(s): 上田 慧、中井万智、志波 智生、高橋 元、 稲岡 ダニエル健、城戸 康年、坂元 君年、 柘植 千明、Emmanuel Oluwadare Balogun、奈良武司、青木孝、本間 光貴、田仲 昭子、井上将行、松岡茂、Moore L. Anthony、斎本 博之、 北 潔、原田 繁春
  • Organizer: 第88回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] Glycerol kinase from human African trypanosomes possess an intrinsic acid phosphatase activity (2014)
  • Author(s): Balogun Emmanuel O., Inaoka Daniel K., Shiba Tomoo, Sakamoto Kimitoshi, Kido Yasutoshi, Nara Takeshi, Aoki Takashi, Honma Teruki, Tanaka Akiko, Inoue Masayuki, Matsuoka Shigeru, Michels Paul AM., Harada Shigeharu, Kita Kiyoshi
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] アフリカトリパノソーマのシアン耐性酸化酵素 (TAO)と新規阻害剤: フェルレノールとの複合体の結晶構造 (2014)
  • Author(s): 上田慧、志波智生、高橋元、稲岡ダニエル健、城戸康年、坂元君年、柘植千明、Emmanuel Oluwadare Balogun、奈良武司、青木孝、本間光貴、田仲昭子、井上将行、松岡茂、Anthony L. Moore、斎本博之、原田繁春、北潔
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] アフリカトリパノソーマのシアン耐性酸化酵素（TAO）と新規阻害剤:フェルレノールとの複合体晶構造 (2014)
  • Author(s): 上田 慧、志波 智生、高橋 元、稲岡 ダニエル健、城戸 康年、坂元 君年、柘植 千明、本間 光貴、田仲 昭子、斎本 博之、北 潔、原田 繁春
  • Organizer: 第１４回日本蛋白質科学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-06-25 – 2014-06-27
• [Book] Cytochrome Complexes: Evolution, Structures, Energy Transduction, and Signaling, Chapter 19, Structure and Mechanism of Action of the Alternative Quinol Oxidases (2016)
  • Author(s): Young L, May B, Shiba T, Harada S, Inaoka DK, Kita K, Moore AL
  • Total Pages: 739
  • Publisher: Springer