In vitro biological evaluation of novel broad-spectrum isothiazolone inhibitors of bacterial type II topoisomerases

Cédric Charrier, Anne-Marie Salisbury, Victoria J. Savage, Emmanuel Moyo, Henry Forward, Nicola Ooi, Jonathan Cheung, Richard Metzger, David McGarry, Rolf Walker, Ian R. Cooper, Andrew J. Ratcliffe, Neil R. Stokes

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Abstract

OBJECTIVES: To evaluate the in vitro biological properties of a novel class of isothiazolone inhibitors of the bacterial type II topoisomerases.

METHODS: Inhibition of DNA gyrase and topoisomerase IV activity was assessed using DNA supercoiling and decatenation assays. MIC and MBC were determined according to CLSI guidelines. Antibacterial combinations were assessed using a two-dimensional chequerboard MIC method. Spontaneous frequency of resistance was measured at various multiples of the MIC. Resistant mutants were generated by serial passage at subinhibitory concentrations of antibacterials and genetic mutations were determined through whole genome sequencing. Mammalian cytotoxicity was evaluated using the HepG2 cell line.

RESULTS: Representative isothiazolone compound REDX04957 and its enantiomers (REDX05967 and REDX05990) showed broad-spectrum bactericidal activity against the ESKAPE organisms, with the exception of Enterococcus spp., as well as against a variety of other human bacterial pathogens. Compounds retained activity against quinolone-resistant strains harbouring GyrA S83L and D87G mutations (MIC ≤4 mg/L). Compounds inhibited the supercoiling activity of wild-type DNA gyrase and the decatenation function of topoisomerase IV. Frequency of resistance of REDX04957 at 4× MIC was <9.1 × 10(-9). Against a panel of recent MDR isolates, REDX05967 demonstrated activity against Acinetobacter baumannii with MIC50 and MIC90 of 16 and 64 mg/L, respectively. Compounds showed a lack of cytotoxicity against HepG2 cells at 128 mg/L.

CONCLUSIONS: Isothiazolone compounds show potent activity against Gram-positive and -negative pathogens with a dual targeting mechanism-of-action and a low potential for resistance development, meriting their continued investigation as broad-spectrum antibacterial agents.

Original languageEnglish
JournalJournal of Antimicrobial Chemotherapy
Early online date26 Jun 2016
DOIs
Publication statusE-pub ahead of print - 26 Jun 2016

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Keywords

  • in vitro
  • isothiazolone inhibitors
  • topoisomerases
  • DNA gyrase
  • mammalian cytotoxicity
  • pathogens
  • antibacterial
  • ESKAPE pathogens
  • anti-infectives
  • isothiazolone

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