Novel insight into the reaction of nitro, nitroso and hydroxylamino benzothiazinones and of benzoxacinones with Mycobacterium tuberculosis DprE1

Research output: Contribution to journalArticlepeer-review


  • Adrian Richter
  • Ines Rudolph
  • Ute Möllmann
  • Kerstin Voigt
  • Chun-Wa Chung
  • Onkar M P Singh
  • Michael Rees
  • Alfonso Mendoza-Losana
  • Robert Bates
  • Lluís Ballell
  • Peter Imming
  • Argyrides Argyrou

Colleges, School and Institutes

External organisations

  • Institut für Pharmazie, Martin-Luther-Universität Halle-Wittenberg, Wolfgang-Langenbeck-Str. 4, 06120, Halle (Saale), Germany.
  • Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie - Hans-Knöll-Institut, Beutenbergstrasse 11a, 07745, Jena, Germany.
  • GlaxoSmithKline
  • Diseases of the Developing World, Tres Cantos Medicines Development Campus, GlaxoSmithKline, Severo Ochoa 2, 28760 Tres Cantos, Madrid, Spain.
  • Institut für Pharmazie, Martin-Luther-Universität Halle-Wittenberg, Wolfgang-Langenbeck-Str. 4, 06120, Halle (Saale), Germany.
  • Discovery Biology, Discovery Sciences, IMED Biotech unit, AstraZeneca, Cambridge, United Kingdom.


Nitro-substituted 1,3-benzothiazinones (nitro-BTZs) are mechanism-based covalent inhibitors of Mycobacterium tuberculosis decaprenylphosphoryl-β-D-ribose-2'-oxidase (DprE1) with strong antimycobacterial properties. We prepared a number of oxidized and reduced forms of nitro-BTZs to probe the mechanism of inactivation of the enzyme and to identify opportunities for further chemistry. The kinetics of inactivation of DprE1 was examined using an enzymatic assay that monitored reaction progress up to 100 min, permitting compound ranking according to kinact/Ki values. The side-chain at the 2-position and heteroatom identity at the 1-position of the BTZs were found to be important for inhibitory activity. We obtained crystal structures with several compounds covalently bound. The data suggest that steps upstream from the covalent end-points are likely the key determinants of potency and reactivity. The results of protein mass spectrometry using a 7-chloro-nitro-BTZ suggest that nucleophilic reactions at the 7-position do not operate and support a previously proposed mechanism in which BTZ activation by a reduced flavin intermediate is required. Unexpectedly, a hydroxylamino-BTZ showed time-dependent inhibition and mass spectrometry corroborated that this hydroxylamino-BTZ is a mechanism-based suicide inhibitor of DprE1. With this BTZ derivative, we propose a new covalent mechanism of inhibition of DprE1 that takes advantage of the oxidation cycle of the enzyme.


Original languageEnglish
Article number13473
Number of pages12
JournalScientific Reports
Issue number1
Early online date7 Sep 2018
Publication statusPublished - 7 Sep 2018