Inhibition of polymorphic MexXY-OprM efflux system in Pseudomonas aeruginosa clinical isolates by Berberine derivatives

The MexXY-OprM multidrug efflux pump (EP) in aminoglycosides resistant Pseudomonas aeruginosa is one of the major resistance mechanisms, which is often overexpressed in strains isolated from pulmonary chronic disease such as cystic fibrosis.[1-3] In this research, we focused on the design of potential efflux pumps inhibitors, targeting MexY, the inner membrane component, in an allosteric site. Berberine[4] has been considered as lead molecule since we previously demonstrated its effectiveness in targeting MexY in laboratory reference strains.[5,6] Since this protein is often present in polymorphic variants in clinical strains, we sequenced and modeled all the mutated forms and we synthesized and evaluated by computational techniques, some berberine derivatives carrying an aromatic functionalization in its 13-C ring position. These compounds were tested in vitro against clinical P. aeruginosa strains for antimicrobial and antibiofilm activity. In conclusion, the results demonstrated the importance of the aromatic moiety functionalization in exerting the EP inhibitory activity in synergy with the aminoglycoside tobramycin. More, we found that aminoacidic composition of MexY in different strains must be considered for predicting potential binding site and affects the different activity of berberine derivatives. Finally, the antibiofilm effect of these new EPIs is promising, particularly for o-CH3-berberine derivative.Unveiling Strategies to Combat Drug Resistance: Our study delves into the battle against drug resistance in aminoglycosides resistant Pseudomonas aeruginosa, focusing on the MexXY-OprM multidrug efflux pump system. We explore the design of novel efflux pump inhibitors, targeting MexY at an identified allosteric site (ALP). Lead compound Berberine and its derivatives show promise in inhibiting the pump and reducing biofilm production, offering new avenues in the fight against chronic pulmonary diseases