Highthrouput system for detrmination of drug metabolism in cats

• Project/Area Number: 24580462
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Clinical veterinary science
• Research Institution: Rakuno Gakuen University
• Principal Investigator: TERAOKA Hiroki 酪農学園大学, 獣医学群, 教授 (50222146)
• Co-Investigator(Kenkyū-buntansha): UCHIDE Tsuyoshi 酪農学園大学, 獣医学群, 教授 (20327456) ; INOUE Hiroki 酪農学園大学, 農食環境学群, 准教授 (90305904)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 薬物代謝酵素 / シトクロームP450 / コンパニオンアニマル / ネコ / 薬物相互作用 / 中毒

Outline of Final Research Achievements

Cytochrome P450s (CYPs) are most important metabolizing enzymes for pharmaceutical drugs in mammals. Although domestic cat is the most common companion animal, the properties of the CYPs are largely unknown. In this study, we developed heterologously expressed feline CYPs in Escherichia coli to generate high-throughput CYPs activity assay systems. CYP2E2 transcripts were most abundant followed by 2A13 in the liver, while CYP3A131 is the major CYP in the small intestine. Suitable fluorescent substrate for high-throuput screening was selected for each CYP subtype. Some antifungal agents and sadatives showed strong inhibition in CYP subtype-specific manner. We could not find any drug that inhibits CYP2E2 in lower concentration. Many drugs inhibited CYP3A131 in relatively higher concentrations than human CYP3A4 and canine CYP3A12. The results suggest the usefulness of this system.

Research Products

• [Journal Article] Identification and functional characterization of novel feline cytochrome P450 2A. (2015)
  • Author(s): Okamatsu G, Komatsu T, Kubota A, Onaga T, Uchide T, Endo D, Kirisawa R, Yin G, Inoue H, Kitazawa T, Uno Y, Teraoka H.
  • Journal Title: Xenobiotica Volume: 未定
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Involvement of COX2-thromboxane pathway in TCDD-induced precardiac edema in developing zebrafish (2014)
  • Author(s): Teraoka H, Okuno Y, Nijoukubo D, Yamakoshi A, Peterson R, Stegeman JJ, Kitazawa T, Hiraga T. and Kubota A.
  • Journal Title: Auat Toxicol. Volume: 154 Pages: 19-26
  • DOI: 10.1016/j.aquatox.2014.04.025
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Mass mortality of Eurasian tree sparrow (Passer montanus) due to emerging epidemic infection of Salmonella Typhimurium DT40 in Japan (2014)
  • Author(s): Fukui D. Takahashi K, Kudo M, Une Y, Katoh Y, Izumiya H, Teraoka H, Asakawa M, Yanagida K, Bando G
  • Journal Title: Journal of Wildlife Diseases Volume: 50巻(印刷中) Issue: 3 Pages: 484-495
  • DOI: 10.7589/2012-12-321
  • Peer Reviewed
• [Journal Article] 動物の毒性物質感受性に関する現在の知見；種差，系統差，個体差 (2014)
  • Author(s): 寺岡宏樹
  • Journal Title: 中毒研究 Volume: 27 Pages: 314-319
• [Journal Article] Vertebral Formula in Red-Crowned Crane (Grus japonensis) and Hooded Crane (Grus monacha) (2014)
  • Author(s): Hiraga T, Sakamoto H, Nishikawa S, Muneuchi I, Ueda H, Inoue M, Shimura R, Uebayashi A, Yasuda N, Momose K, Masatomi H, Teraoka H
  • Journal Title: J Vet Med Sci. Volume: 76 Pages: 503-508
  • NAID: 130003382404
  • Peer Reviewed
• [Journal Article] Involvement of COX2–thromboxane pathway in TCDD-induced precardiac edema in developing zebrafish (2014)
  • Author(s): Teraoka H, Okuno Y, Nijoukubo D, Yamakoshi Y, et al.
  • Journal Title: Aquat Toxicol Volume: in press
  • Peer Reviewed
• [Journal Article] Cyp1a reporter zebrafish reveals target tissues for dioxin (2013)
  • Author(s): Kim KH, Park HJ, Kim JH, Kim S, Williams DR, Kim MK, Jung YD, Teraoka H, Park HC, Choy HE, Shin BA, Choi SY.
  • Journal Title: Aquat Toxicol. 2013 Jun 15;134-135:57-65. Volume: 134-135 Pages: 57-65
  • DOI: 10.1016/j.aquatox.2013.03.010
  • Peer Reviewed
• [Journal Article] Age-dependent reduction of ghrelin- and motilin-induced contractile activity in the chicken gastrointestinal tract. (2013)
  • Author(s): Kitazawa, T. et al.
  • Journal Title: peptides Volume: 43 Pages: 88-95
  • DOI: 10.1016/j.peptides.2013.02.012
  • Peer Reviewed
• [Journal Article] Large-Scale Survey of Mitochondrial D-Loop of the Red-Crowned Crane <i>Grus japonensis</i> in Hokkaido, Japan by Convenient Genotyping Method (2013)
  • Author(s): Miura Y, Shiomi A, Shiraishi J, Makita K, Asakawa M, Kitazawa T, Hiraga T, Momose Y, Momose K, Masatomi H, Teraoka H.
  • Journal Title: Journal of Veterinary Medical Science Volume: 75 Issue: 1 Pages: 43-47
  • DOI: 10.1292/jvms.12-0193
  • NAID: 130001879690
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] Physical Body Parameters of Red-Crowned Cranes Grus japonensis by Sex and Life Stage in Eastern Hokkaido, Japan. (2013)
  • Author(s): Inoue M, Shimura R, Uebayashi A, Ikoma S, Iima H, Sumiyoshi T, Teraoka H, Makita K, Hiraga T, Momose K, Masatomi H.
  • Journal Title: Journal of Veterinary Medical Sciences Volume: 印刷中
  • NAID: 130003362179
  • Peer Reviewed
• [Journal Article] Origin of Three Red-Crowned Cranes Grus japonensis Found in Northeast Honshu and West Hokkaido, Japan, from 2008 to 2012. (2013)
  • Author(s): Miura Y, Shiraishi J, Shiomi A, Kitazawa T, Hiraga T, Matsumoto F, Teraoka H, Masatomi H.
  • Journal Title: Journal of Veterinary Medical Sciences Volume: 印刷中
  • NAID: 130003362199
  • Peer Reviewed
• [Presentation] ネコ シトクロームP450発現系の構築と阻害薬によるスクリーニング (2014)
  • Author(s): 岡松 岳、小松 徹也、宇野 泰広、打出毅、北澤 多喜雄、寺岡宏樹
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜、横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Chicken is a spcific and useful animal model to study the functional role of ghrelin and motilin in regulation of gastrointasitinal motility (2014)
  • Author(s): Kitazawa T , Teraoka H , Kaiya H
  • Organizer: 20th International Symposium on Regulatory Peptides
  • Place of Presentation: Kyoto Garden Palace, Kyoto, Japan
  • Year and Date: 2014-09-07 – 2014-09-10
• [Presentation] Interactions between thromboxane and prostacyclin in precardial edema formation in developing zebrafish exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin
  • Author(s): Hiroki Teraoka, Yuki Okuno, Ayumi Yamakoshi, Kengo Sasano, Richard E. Peterson, Takio Kitazawa, Takeo Hiraga and Akira Kubota
  • Organizer: The XIII International Congress of Toxicology
  • Place of Presentation: COEX, Seoul, Korea
• [Presentation] ネコ、イヌ、ヒトにおけるシトクローム P450 3A分子種の動物種差
  • Author(s): 岡松 岳、川上 圭、小松徹也、宇野泰広、北澤多喜雄、寺岡宏樹
  • Organizer: 第28回日本中毒学会東日本地方会
  • Place of Presentation: 北里大学薬学部、東京都
• [Presentation] Comparative Characterization of Feline Cytochrome P450
  • Author(s): Hiroki Teraoka, Gaku Okamatsu, Tetsuya Komatsu and Takio Kitazawa
  • Organizer: 54th Society of Toxicology, Annual Meeting
  • Place of Presentation: Phoenix Convention Center, Phoenix, AZ, USA
• [Presentation] 発達初期ゼブラフィッシュにおける循環障害の発現機構
  • Author(s): 寺岡宏樹
  • Organizer: 第39回 日本毒性学会学術年会
  • Place of Presentation: 仙台国際会議場
  • Invited
• [Presentation] 大腸菌発現系を用いたネコ シトクローム P450 3Aのハイスループット薬物相互作用スクリーニング系の構築
  • Author(s): 寺岡宏樹、川上 圭、浅倉弘幸、岡松 岳、井上博紀、宇野泰広、北澤多喜雄
  • Organizer: 第155回 日本獣医学会学術年会
  • Place of Presentation: 東京大学駒場キャンパス