Platform to Discover Protease-Activated Antibiotics and Application to Siderophore-Antibiotic Conjugates.

ABSTRACT: Here we present a platform for discovery of protease-activated prodrugs and apply it to antibiotics that target Gram-negative bacteria. Because cleavable linkers for prodrugs had not been developed for bacterial proteases, we used substrate phage to discover substrates for proteases found in the bacterial periplasm. Rather than focusing on a single protease, we used a periplasmic extract of E. coli to find sequences with the greatest susceptibility to the endogenous mixture of periplasmic proteases. Using a fluorescence assay, candidate sequences were evaluated to identify substrates that release native amine-containing payloads. We next designed conjugates consisting of (1) an N-terminal siderophore to facilitate uptake, (2) a protease-cleavable linker, and (3) an amine-containing

SUBMITTER: Boyce JH 

PROVIDER: S-EPMC7790458 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature