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

摘要: Here we present a platform for discovery of protease-activated prodrugs and apply it to antibiotics that target Gram-negative bacteria. Because cleavable linkers had not been developed bacterial proteases, used substrate phage discover substrates proteases found in the periplasm. Rather than focusing on single protease, periplasmic extract E. coli find sequences with greatest susceptibility endogenous mixture proteases. Using fluorescence assay, candidate were evaluated identify release native amine-containing payloads. We next designed conjugates consisting (1) an N-terminal siderophore facilitate uptake, (2) protease-cleavable linker, (3) antibiotic. this strategy, converted daptomycin-which by itself is active only against Gram-positive bacteria-into antibiotic capable targeting Acinetobacter species. similarly demonstrated siderophore-facilitated delivery oxazolidinone macrolide into number These results illustrate platform's utility development prodrugs, including Trojan horse antibiotics.