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Discovery and chemical optimisation of a Potent, Bi-cyclic (Bicycle(R)) Antimicrobial Inhibitor of Escherichia coli PBP3

Rowland, C. E.; Newman, H.; Martin, T. T.; Dods, R.; Bournakas, N.; Wagstaff, J. M.; Lewis, N.; Stanway, S. J.; Balmforth, M.; Kessler, C.; van Rietschoten, K.; Bellini, D.; Roper, D. I.; Lloyd, A. J.; Dowson, C. G.; Skynner, M. J.; Beswick, P.; Dawson, M. J.

2024-03-21 biochemistry
10.1101/2024.03.20.581580 bioRxiv
Show abstract

Penicillin binding proteins (PBPs) are well validated antimicrobial targets, but the prevalence of {beta}-lactamase driven resistance and, more rarely, target-based mutations, necessitates new classes of PBP-targeting drugs. Here we describe the discovery and optimisation of novel, bicyclic peptide (Bicycle(R)) inhibitors of E. coli PBP3 (EcPBP3) using a proprietary phage display platform, and their conjugation to linear antimicrobial peptides to confer outer membrane permeation. These molecules exhibited high-affinity binding to E. coli PBP3 and a viable spectrum of killing activity against clinically relevant species of the Enterobacterales. X-ray crystallography was used to explore the mode of binding to PBP3, enabling increased target affinity and improvement of in vitro stability. These compounds bind to the transpeptidase active site cleft of PBP3 and represent a novel non-{beta}-lactam chemical class of high affinity, non-covalent penicillin binding protein inhibitors.

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