ZN148 Is a Modular Synthetic Metallo-β-Lactamase Inhibitor That Reverses Carbapenem Resistance in Gram-Negative Pathogens In Vivo

  • Antimicrob Agents Chemother. 2020 May 21;64(6):e02415-19. doi: 10.1128/AAC.02415-19.
Ørjan Samuelsen  1  2 ,  Ove Alexander Høgmoen Åstrand  3 ,  Christopher Fröhlich  4  5 ,  Adam Heikal  6 ,  Susann Skagseth  5 ,  Trine Josefine Olsen Carlsen  5 ,  Hanna-Kirsti S Leiros  5 ,  Annette Bayer  7 ,  Christian Schnaars  3 ,  Geir Kildahl-Andersen  3 ,  Silje Lauksund  4  2 ,  Sarah Finke  6 ,  Sandra Huber  8 ,  Tor Gjøen  6 ,  Adriana Magalhaes Santos Andresen  6 ,  Ole Andreas Økstad  6 ,  Pål Rongved  9
Affiliations
  • 1. Norwegian National Advisory Unit on Detection of Antimicrobial Resistance, Department of Microbiology and Infection Control, University Hospital of North Norway, Tromsø, Norway [email protected] [email protected].
  • 2. Department of Pharmacy, UiT The Arctic University of Norway, Tromsø, Norway.
  • 3. Section for Pharmaceutical Chemistry, Department of Pharmacy, University of Oslo, Oslo, Norway.
  • 4. Norwegian National Advisory Unit on Detection of Antimicrobial Resistance, Department of Microbiology and Infection Control, University Hospital of North Norway, Tromsø, Norway.
  • 5. The Norwegian Structural Biology Centre (NorStruct), Department of Chemistry, UiT The Arctic University of Norway, Tromsø, Norway.
  • 6. Centre for Integrative Microbial Evolution and Section for Pharmacology and Pharmaceutical Biosciences, Department of Pharmacy, University of Oslo, Oslo, Norway.
  • 7. Department of Chemistry, UiT The Arctic University of Norway, Tromsø, Norway.
  • 8. Department of Laboratory Medicine, Division of Diagnostic Services, University Hospital of North Norway, Tromsø, Norway.
  • 9. Section for Pharmaceutical Chemistry, Department of Pharmacy, University of Oslo, Oslo, Norway [email protected] [email protected].
PMID: 32179522 DOI: 10.1128/AAC.02415-19
Abstract

Carbapenem-resistant Gram-negative pathogens are a critical public health threat and there is an urgent need for new treatments. Carbapenemases (β-lactamases able to inactivate carbapenems) have been identified in both serine β-lactamase (SBL) and Metallo-β-lactamase (MBL) families. The recent introduction of SBL carbapenemase inhibitors has provided alternative therapeutic options. Unfortunately, there are no approved inhibitors of MBL-mediated carbapenem-resistance and treatment options for infections caused by MBL-producing Gram-negatives are limited. Here, we present ZN148, a zinc-chelating MBL-inhibitor capable of restoring the bactericidal effect of meropenem and in vitro clinical susceptibility to carbapenems in >98% of a large international collection of MBL-producing clinical Enterobacterales strains (n = 234). Moreover, ZN148 was able to potentiate the effect of meropenem against NDM-1-producing Klebsiella pneumoniae in a murine neutropenic peritonitis model. ZN148 showed no inhibition of the human zinc-containing enzyme glyoxylase II at 500 μM, and no acute toxicity was observed in an in vivo mouse model with cumulative dosages up to 128 mg/kg. Biochemical analysis showed a time-dependent inhibition of MBLs by ZN148 and removal of zinc ions from the active site. Addition of exogenous zinc after ZN148 exposure only restored MBL activity by ∼30%, suggesting an irreversible mechanism of inhibition. Mass-spectrometry and molecular modeling indicated potential oxidation of the active site Cys221 residue. Overall, these results demonstrate the therapeutic potential of a ZN148-carbapenem combination against MBL-producing Gram-negative pathogens and that ZN148 is a highly promising MBL inhibitor that is capable of operating in a functional space not presently filled by any clinically approved compound.

Keywords
antibiotic resistance; carbapenem; metallo-β-lactamase; β-lactamase inhibitor; β-lactamases.
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