Author:
Eakin Ann E.,Green Oluyinka,Hales Neil,Walkup Grant K.,Bist Shanta,Singh Alok,Mullen George,Bryant Joanna,Embrey Kevin,Gao Ning,Breeze Alex,Timms Dave,Andrews Beth,Uria-Nickelsen Maria,Demeritt Julie,Loch James T.,Hull Ken,Blodgett April,Illingworth Ruth N.,Prince Bryan,Boriack-Sjodin P. Ann,Hauck Sheila,MacPherson Lawrence J.,Ni Haihong,Sherer Brian
Abstract
ABSTRACTDNA gyrase is an essential enzyme in bacteria, and its inhibition results in the disruption of DNA synthesis and, subsequently, cell death. The pyrrolamides are a novel class of antibacterial agents targeting DNA gyrase. These compounds were identified by a fragment-based lead generation (FBLG) approach using nuclear magnetic resonance (NMR) screening to identify low-molecular-weight compounds that bind to the ATP pocket of DNA gyrase. A pyrrole hit with a binding constant of 1 mM formed the basis of the design and synthesis of a focused library of compounds that resulted in the rapid identification of a lead compound that inhibited DNA gyrase with a 50% inhibitory concentration (IC50) of 3 μM. The potency of the lead compound was further optimized by utilizing iterative X-ray crystallography to yield DNA gyrase inhibitors that also displayed antibacterial activity. Spontaneous mutants were isolated inStaphylococcus aureusby plating on agar plates containing pyrrolamide 4 at the MIC. The resistant variants displayed 4- to 8-fold-increased MIC values relative to the parent strain. DNA sequencing revealed two independent point mutations in the pyrrolamide binding region of thegyrBgenes from these variants, supporting the hypothesis that the mode of action of these compounds was inhibition of DNA gyrase. Efficacy of a representative pyrrolamide was demonstrated againstStreptococcus pneumoniaein a mouse lung infection model. These data demonstrate that the pyrrolamides are a novel class of DNA gyrase inhibitors with the potential to deliver future antibacterial agents targeting multiple clinical indications.
Publisher
American Society for Microbiology
Subject
Infectious Diseases,Pharmacology (medical),Pharmacology
Cited by
95 articles.
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