Whole Cell Target Engagement Identifies Novel Inhibitors of Mycobacterium tuberculosis Decaprenylphosphoryl-β-d-ribose Oxidase

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• 作者：Sarah M. Batt ; 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 ; Gurdyal S. Besra ; Argyrides Argyrou
• 刊名：ACS Infectious Diseases
• 出版年：2015
• 出版时间：December 11, 2015
• 年：2015
• 卷：1
• 期：12
• 页码：615-626
• 全文大小：697K
• ISSN：2373-8227

文摘

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.