Whole cell target engagement identifies novel inhibitors of Mycobacterium tuberculosis decaprenylphosphoryl-β-D-ribose oxidase

Research output: Contribution to journalArticlepeer-review

Authors

  • Monica Cacho Izquierdo
  • Julia Castro Pichel
  • Christopher J Stubbs
  • Laura Vela-Glez Del Peral
  • Esther Pérez-Herrán
  • Neeraj Dhar
  • Bernadette Mouzon
  • Mike Rees
  • Jonathan P Hutchinson
  • Robert J Young
  • John D McKinney
  • David Barros Aguirre
  • Lluis Ballell
  • Argyrides Argyrou

Colleges, School and Institutes

Abstract

We have targeted the Mycobacterium tuberculosis decaprenylphosphoryl-β-D-ribose oxidase (Mt-DprE1) for potential chemotherapeutic intervention of tuberculosis. A multicopy suppression strategy that overexpressed Mt-DprE1 in M. bovis BCG was used to profile the publically available GlaxoSmithKline antimycobacterial compound set, and one compound (GSK710) was identified that showed an 8-fold higher minimum inhibitory concentration relative to the control strain. Analogues of GSK710 show a clear relationship between whole cell potency and in vitro activity using an enzymatic assay employing recombinant Mt-DprE1, with binding affinity measured by fluorescence quenching of the flavin cofactor of the enzyme. M. bovis BCG spontaneous resistant mutants to GSK710 and a closely related analogue were isolated and sequencing of ten such mutants revealed a single point mutation at two sites, E221Q or G248S within DprE1, providing further evidence that DprE1 is the main target of these compounds. Finally, time-lapse microscopy experiments showed that exposure of M. tuberculosis to a compound of this series arrests bacterial growth rapidly followed by a slower cytolysis phase.

Details

Original languageEnglish
Pages (from-to)615-626
Number of pages12
JournalACS Infectious Diseases
Volume1
Issue number12
Early online date25 Aug 2015
Publication statusPublished - 11 Dec 2015

Keywords

  • multicopy suppression, target overexpression, time-lapse microscopy, drug discovery

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