新規の肝臓ヒト化薬物性肝障害モデルマウスによるヒト特異的薬物肝毒性の発現機序解明

2022 Fiscal Year Annual Research Report

• Project/Area Number: 20K06463
• Research Institution: Central Institute for Experimental Animals
• Principal Investigator: 上原 正太郎 公益財団法人実験動物中央研究所, 実験動物応用研究部, 研究員 (10733123)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: ヒト化肝臓マウス / チトクロムP450 / チトクロムP450酸化還元酵素 / POR / TK-NOGマウス / S-ワルファリン / 薬物代謝 / ヒト型薬物代謝

Outline of Annual Research Achievements

最終年度はS-ワルファリンを用いたin vivo代謝実験によりPor cKO肝臓ヒト化マウスの薬物代謝能を検証した。S-ワルファリンを単回静脈内投与後のPor野生型マウス、Por cKO（null/flox）マウス、従来型ヒト化肝臓マウスおよびPor cKOヒト化肝臓マウスの血漿および尿中のS-ワルファリン代謝物（S-4'-/6-/7-/8-水酸化ワルファリン）を測定した。代謝物の血中濃度曲線下面積を比較すると、4'-水酸化体はPOR cKOヒト化肝臓マウスの方が従来型ヒト化肝臓マウスに比べて低く、6または7-水酸化体は従来型ヒト化肝臓マウスよりもPor cKOヒト化肝臓マウスの方が高かった。S-ワルファリン投与後のPor cKOヒト化肝臓マウスの尿中に最も多い代謝物はS-7-水酸化体であり、次いでS-6-水酸化体、S-4'-水酸化体および8-水酸化体であった。マウスのS-ワルファリン主要代謝物であるS-4'-水酸化体は、Por cKOヒト化肝臓マウス尿中の従来型ヒト化肝臓マウスに比べて著しく少なく、肝臓のPorが欠損した特徴を反映していることが推察された。以上、Por cKOヒト化肝臓マウスは、肝臓の残存マウスP450酵素活性が低減したことにより、従来型ヒト肝キメラマウスと比べてより鮮明にヒト型のS-ワルファリン代謝を示した。研究成果を統括すると、従来型ヒト化肝臓モデルは残存マウス肝P450活性が原因で医薬候補品の有効性および安全性の予測不良を招く可能性があった。本研究で作製したPor cKOヒト化肝臓マウスは肝臓特異的Por欠損に伴ってマウス肝P450酵素活性が低下しており、従来型のヒト化肝臓モデルに比べ、よりヒトに近い薬物代謝能を持つことが示唆された。今後、肝Por欠損ヒト化肝臓マウスは薬物動態・安全性研究のための新しいプラットフォームとしてその活用が期待される。

Research Products

• [Journal Article] The Unique Human N10-Glucuronidated Metabolite Formation from Olanzapine in Chimeric NOG-TKm30 Mice with Humanized Livers. (2023)
  • Author(s): Uehara S, Higuchi Y, Yoneda N, Kato H, Yamazaki H, Suemizu H.
  • Journal Title: Drug Metab Dispos. Volume: 51 Pages: 480-491
  • DOI: 10.1124/dmd.122.001102.
  • Peer Reviewed
• [Journal Article] Drug transporter expression and activity in cryopreserved human hepatocytes isolated from chimeric TK-NOG mice with humanized livers. (2023)
  • Author(s): Zerdoug A, Le Vee M, Uehara S, Jamin A, Higuchi Y, Yoneda N, Lopez B, Chesne C, Suemizu H, Fardel O.
  • Journal Title: Toxicol In Vitro. Volume: 6 Pages: 105592
  • DOI: 10.1016/j.tiv.2023.105592.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Roles of human cytochrome P450 3A4/5 in dexamethasone 6β-hydroxylation mediated by liver microsomes and humanized liver in chimeric mice metabolically suppressed with azamulin. (2023)
  • Author(s): Uehara S, Shimizu M, Suemizu H, Yamazaki H.
  • Journal Title: Drug Metab Pharmacokinet. Volume: 50 Pages: 100504
  • DOI: 10.1016/j.dmpk.2023.100504.
  • Peer Reviewed
• [Journal Article] Cytochrome P450-dependent drug oxidation activities and their expression levels in liver microsomes of chimeric TK-NOG mice with humanized livers. (2022)
  • Author(s): Uehara S, Yoneda N, Higuchi Y, Yamazaki H, Suemizu H.
  • Journal Title: Drug Metab Pharmacokinet. Volume: 44 Pages: 100454
  • DOI: 10.1016/j.dmpk.2022.100454.
  • Peer Reviewed
• [Journal Article] Probe drug T-1032 N-oxygenation mediated by cytochrome P450 3A5 in human hepatocytes in vitro and in humanized-liver mice in vivo. (2022)
  • Author(s): Uehara S, Shimizu M, Ple K, Routier S, Yoneda N, Higuchi Y, Suemizu H, Yamazaki H.
  • Journal Title: Drug Metab Pharmacokinet. Volume: 44 Pages: 100453
  • DOI: 10.1016/j.dmpk.2022.100453.
  • Peer Reviewed
• [Journal Article] Comparison of mouse and human cytochrome P450 mediated-drug metabolising activities in hepatic and extrahepatic microsomes. (2022)
  • Author(s): Uehara S, Murayama N, Higuchi Y, Yoneda N, Yamazaki H, Suemizu H.
  • Journal Title: Xenobiotica Volume: 52 Pages: 229-239
  • DOI: 10.1080/00498254.2022.2066581.
  • Peer Reviewed
• [Journal Article] Humanized liver TK-NOG mice with functional deletion of hepatic murine cytochrome P450s as a model for studying human drug metabolism. (2022)
  • Author(s): Uehara S, Iida Y, Ida-Tanaka M, Goto M, Kawai K, Yamamoto M, Higuchi Y, Ito S, Takahashi R, Kamimura H, Ito M, Yamazaki H, Oshimura M, Kazuki Y, Suemizu H.
  • Journal Title: Sci Rep. Volume: 12 Pages: 14907
  • DOI: 10.1038/s41598-022-19242-0.
  • Peer Reviewed / Open Access
• [Journal Article] Contribution of Humanized Liver Chimeric Mice to the Study of Human Hepatic Drug Transporters: State of the Art and Perspectives. (2022)
  • Author(s): Zerdoug A, Le Vee M, Uehara S, Lopez B, Chesne C, Suemizu H, Fardel O.
  • Journal Title: Eur J Drug Metab Pharmacokinet. Volume: 47 Pages: 621-637
  • DOI: 10.1007/s13318-022-00782-9.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Cytochrome P450s in chimeric mice with humanized liver. (2022)
  • Author(s): Uehara S, Suemizu H, Yamazaki H.
  • Journal Title: Adv Pharmacol. Volume: 95 Pages: 307-328
  • DOI: 10.1016/bs.apha.2022.05.004.
  • Peer Reviewed
• [Journal Article] Expression and functional activity of cytochrome P450 enzymes in human hepatocytes with sustainable reproducibility for in vitro phenotyping studies. (2022)
  • Author(s): Bachour-El Azzi P, Chesne C, Uehara S.
  • Journal Title: Adv Pharmacol. Volume: 95 Pages: 285-305
  • DOI: 10.1016/bs.apha.2022.05.009.
  • Peer Reviewed / Int'l Joint Research
• [Presentation] 創薬への応用を指向した新規ヒト肝細胞移植マウスの開発と薬物代謝能評価 (2023)
  • Author(s): 上原正太郎
  • Organizer: 第5回 日本獣医薬理学・毒性学会 春季研究会
  • Invited
• [Presentation] Conditional knockout of the cytochrome P450 oxidoreductase gene highlights human drug metabolism in humanized liver TK-NOG mice (2022)
  • Author(s): Suemizu H, Iida Y, Ida-Tanaka M, Kawai K, Higuchi Y, Ito S, Kamimura H, Ito M, Yamazaki H, Oshimura M, Kazuki Y, Uehara S.
  • Organizer: ISSX/MDO 2022 MEETING
  • Int'l Joint Research
• [Presentation] Characterization of drug-metabolizing enzymatic activities in the hepatocytes isolated from the chimeric TK-NOG mice with humanized livers (2022)
  • Author(s): Uehara S, Higuchi Y, Yoneda N, Murayama N, Yamazaki H, Suemizu H.
  • Organizer: ISSX/MDO 2022 MEETING
  • Int'l Joint Research
• [Presentation] Development of cytochrome P450 oxidoreductase (POR) knockout TK-NOG humanized liver chimeric mice for studying drug metabolism research in humans (2022)
  • Author(s): Suemizu H, Iida Y, Ida-Tanaka M, Kawai K, Higuchi Y, Ito S, Kamimura H, Ito M, Yamazaki H, Oshimura M, Kazuki Y, Uehara S.
  • Organizer: 日本薬物動態学会第37回年会
• [Presentation] Unique human 10N-glucuronidated metabolite formation from olanzapine in chimeric NOG-TKm30 mice transplanted with human hepatocytes (2022)
  • Author(s): Uehara S, Higuchi Y, Yoneda N, Yamazaki H, Suemizu H.
  • Organizer: 日本薬物動態学会第37回年会