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The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions
08:06

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Published on: February 1, 2018

Assay platform for clinically relevant metallo-β-lactamases.

Sander S van Berkel1, Jürgen Brem, Anna M Rydzik

  • 1Chemistry Research Laboratory, University of Oxford , 12 Mansfield Road, Oxford OX1 3TA, United Kingdom.

Journal of Medicinal Chemistry
|August 1, 2013
PubMed
Summary

Metallo-β-lactamases (MBLs) pose a significant threat to antibiotics. This study developed new fluorogenic substrates and identified potential pan-MBL inhibitors, 4-chloroisoquinolinols, to combat MBL resistance.

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Area of Science:

  • Microbiology and Infectious Diseases
  • Drug Discovery and Development
  • Biochemistry

Background:

  • Metallo-β-lactamases (MBLs) are critical enzymes conferring resistance to β-lactam antibiotics.
  • The rise of MBLs necessitates the development of novel inhibitors and effective screening platforms.
  • Existing screening methods often lack the sensitivity and scope required for broad-spectrum MBL inhibitor discovery.

Purpose of the Study:

  • To establish a robust screening platform for identifying metallo-β-lactamase inhibitors.
  • To develop and validate novel fluorogenic substrates for MBL activity detection.
  • To identify potential pan-MBL inhibitors effective against clinically relevant MBL variants.

Main Methods:

  • Preparation of clinically relevant MBLs: NDM-1, IMP-1, SPM-1, and VIM-2.
  • Synthesis and characterization of umbelliferone-derived fluorogenic cephalosporin substrates.
  • Comparative analysis of fluorogenic substrates against chromogenic substrates (CENTA, nitrocefin, imipenem).
  • Inhibitor screening using the developed fluorogenic substrate platform.

Main Results:

  • Successfully prepared a panel of clinically significant MBLs.
  • Developed sensitive fluorogenic substrates with improved kinetic parameters compared to chromogenic alternatives.
  • Identified 4-chloroisoquinolinols as potent inhibitors with potential pan-MBL activity.
  • Demonstrated the utility of the fluorogenic assay for efficient inhibitor screening.

Conclusions:

  • The developed fluorogenic substrate assay provides a sensitive and efficient platform for MBL inhibitor discovery.
  • Umbelliferone-derived cephalosporins are effective substrates for screening MBLs.
  • 4-chloroisoquinolinols represent promising candidates for broad-spectrum MBL inhibitors, addressing a critical unmet medical need.