Discovery of 3-hydroxypyridin-4(1H)-ones ester of ciprofloxacin as prodrug to combat biofilm-associated Pseudomonas aeruginosa

Abstract

Chronic infections by Pseudomonas aeruginosa (P. aeruginosa) are frequently complicated due to its ability to form biofilm, which also effectively enhance its resistance to antibiotics. Bacteria-specific antibiotic delivery could locally increase drug concentration to break antimicrobial resistance and reduce the drug's peripheral side effects. The standard-of-care drug ciprofloxacin suffers from severe systemic side effects and was therefore chosen for this approach. It has been identified that 3-hydroxypyridin-4(1H)-one as siderophore mimics could be utilized by P. aeruginosa, and reduced bacterial biofilm formation. In this work, ciprofloxacin was conjugated to 3-hydroxypyridin-4(1H)-one by cleavable linkers to yield prodrugs, which were strategically designed and synthesized to function as dual antibacterial and antibiofilm agents against P. aeruginosa. Conjugate 5c was identified and has the best minimum inhibitory concentrations of 1.07 μM against P. aeruginosa PAO1, and reduced 61.7 % of biofilm formation. In addition, 5c destroyed 75.7 % of mature biofilms. Further studies on the uptake mechanisms showed that the bacterial siderophore-dependent iron transport system was involved in the uptake of the conjugates. Conjugate 5c interfered with iron uptake by bacteria, inhibited their motilities and reduced the production of virulence. Furthermore, prodrug 5c reduced toxicity in vivo and in vitro and showed a positive therapeutic effect in the treatment of Caenorhabditis elegans (C. elegans) infected by P. aeruginosa. These results demonstrate that 3-hydroxypyridin-4(1H)-ones-ciprofloxacin prodrugs are potent in the treatment of biofilm-associated drug-resistant P. aeruginosa infections.