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Identification of 2‑Aryl-Quinolone Inhibitors of Cytochrome bd and Chemical Validation of Combination Strategies for Respiratory Inhibitors against Mycobacterium tuberculosis
journal contributionposted on 2023-01-06, 10:47 authored by Laura N. Jeffreys, Alison Ardrey, Taghreed A. Hafiz, Lauri-Anne Dyer, Ashley J. Warman, Nada Mosallam, Gemma L. Nixon, Nicholas E. Fisher, W. David Hong, Suet C. Leung, Ghaith Aljayyoussi, Jaclyn Bibby, Deepak V. Almeida, Paul J. Converse, Nader Fotouhi, Neil G. Berry, Eric L. Nuermberger, Anna M. Upton, Paul M. O’Neill, Stephen A. Ward, Giancarlo A. Biagini
Mycobacterium tuberculosis cytochrome bd quinol oxidase (cyt bd), the alternative terminal oxidase of the respiratory chain, has been identified as playing a key role during chronic infection and presents a putative target for the development of novel antitubercular agents. Here, we report confirmation of successful heterologous expression of M. tuberculosis cytochrome bd. The heterologous M. tuberculosis cytochrome bd expression system was used to identify a chemical series of inhibitors based on the 2-aryl-quinolone pharmacophore. Cytochrome bd inhibitors displayed modest efficacy in M. tuberculosis growth suppression assays together with a bacteriostatic phenotype in time-kill curve assays. Significantly, however, inhibitor combinations containing our front-runner cyt bd inhibitor CK-2-63 with either cyt bcc-aa3 inhibitors (e.g., Q203) and/or adenosine triphosphate (ATP) synthase inhibitors (e.g., bedaquiline) displayed enhanced efficacy with respect to the reduction of mycobacterium oxygen consumption, growth suppression, and in vitro sterilization kinetics. In vivo combinations of Q203 and CK-2-63 resulted in a modest lowering of lung burden compared to treatment with Q203 alone. The reduced efficacy in the in vivo experiments compared to in vitro experiments was shown to be a result of high plasma protein binding and a low unbound drug exposure at the target site. While further development is required to improve the tractability of cyt bd inhibitors for clinical evaluation, these data support the approach of using small-molecule inhibitors to target multiple components of the branched respiratory chain of M. tuberculosis as a combination strategy to improve therapeutic and pharmacokinetic/pharmacodynamic (PK/PD) indices related to efficacy.
novel antitubercular agentsmycobacterium oxygen consumptionkill curve assaysg ., bedaquilinelung burden comparedalternative terminal oxidasetarget multiple componentsg ., q203displayed enhanced efficacybranched respiratory chain3 subsuccessful heterologous expressioninhibitor combinations containing2 ‑ arylrespiratory chainquinol oxidaseexpression system>< subtarget siteputative targetrespiratory inhibitorsvivo vitro bd bcc aa using smallsterilization kineticsreport confirmationreduced efficacyquinolone pharmacophoreq203 alonemodest loweringkey roleinhibitor ckindices relatedgrowth suppressionexperiments compareddata supportcombination strategycombination strategiesclinical evaluationchronic infectionchemical validationchemical seriesbacteriostatic phenotypeadenosine triphosphate