Development of irreversible inhibitors forβ-lactam inactived enzyme, metallo-β-lactamase

2011 Fiscal Year Final Research Report

• Project/Area Number: 21590116
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Drug development chemistry
• Research Institution: Kumamoto University
• Principal Investigator: KUROSAKI Hiromasa 熊本大学, 大学院・生命科学研究部, 准教授 (70234599)
• Co-Investigator(Kenkyū-buntansha): YAMAGUCHI Yoshihiro 熊本大学, 環境安全センター, 准教授 (10363524)
• Co-Investigator(Renkei-kenkyūsha): ARAKAWA Yoshichika 名古屋大学, 大学院・医学系研究科, 教授 (10212622)
• Project Period (FY): 2009 – 2011
• Keywords: 医薬分子設計 / 感染症 / β-ラクタム剤 / 薬剤耐性菌 / 阻害剤 / 金属酵素

Research Abstract

IMP-1 metallo-β-lactamase is a dinuclear Zn(II) enzyme that catalyzes the hydrolysis and inactivation of mostβ-lactams including carbapenems, and is involved in one of the mechanisms for generating clinical resistance to antibiotics in pathogenic bacteria. We investigated the metal preferences of Zn(II) and Co(II) for the apo-enzyme of IMP-1 metallo-β-lactamase, apo-IMP-1, which contains a dinuclear metal binding site(the Zn1 and Zn2 sites), by UV-visible spectroscopy. The UV-visible spectrum of apo-IMP-1 containing 1 equiv. of Co(II) and 1 equiv. of Zn(II) showed a high preference of Zn(II) for the Zn1 site compared to Co(II). The interaction of IMP-1 metallo-β-lactamase with mercaptoacetic acid was also investigated using Co(II)-substituted IMP-1 and UV-visible spectroscopy. Possible metal binding modes of Co(II) or Zn(II) to the dinucelar metal binding site in apo-IMP-1 and of mercaptoacetic acid to Co(II)-substituted IMP-1 are proposed.

Research Products

• [Journal Article] A demetallation method for IMP-1 metallo-β-lactamase that restored enzymatic activity upon addition of metal ion(s) (2011)
  • Author(s): Yoshihiro Yamaguchi, Shijia Ding, Emi Murakami, Kayo Imamura, Sachiko Fuchigami, Ryo Hashiguchi, Katsuhide Yutani, Hiromasa Mori, Shinnichiro Suzuki, Yoshichika Arakawa, Hiromasa Kurosaki
  • Journal Title: ChemBioChem Volume: 12(13) Pages: 1979-1983
  • Peer Reviewed
• [Journal Article] Metal preference of Zn(II) and Co(II) for the dinuclear metal binding site of IMP-1 metallo-β-lactamase and spectroscopic properties of Co(II)-substituted IMP-1 with mercaptoacetic acid (2011)
  • Author(s): Yoshihiro Yamaguchi, Kayo Imamura, Ako Sasao, Emi Murakami, Yoshichika Arakawa, Hiromasa Kurosaki
  • Journal Title: Med. Chem. Commun Volume: 2(8) Pages: 720-725
  • Peer Reviewed
• [Presentation] Detection of metallo-β-lactamase producing bacteria by fluoresent probe having dansyl and thiol groups (2010)
  • Author(s): Kouki Matsuda, Yoshihiro Yamaguchi, Junzo Hirose, Yoshichika Arakawa, Hiromasa Kurosaki
  • Organizer: 第20回金属の関与する生体関連反応シンポジウム
  • Place of Presentation: 徳島文理大学(徳島)
  • Year and Date: 2010-06-25
• [Presentation] 薬剤耐性菌が産生するメタロ-β-ラクタマーゼの蛍光検出法の開発:ダンシルとチオール基を持つ蛍光剤のメタロ-β-ラクタマーゼIMP-1, VIM-2, IND-7に対する蛍光特性と速度論解析 (2009)
  • Author(s): 松田幸喜、山口佳宏、廣瀬順三、荒川宜親、黒崎博雅
  • Organizer: 第38回薬剤耐性菌研究会
  • Place of Presentation: 森秋旅館(群馬)
  • Year and Date: 2009-11-20