Structural analysis and new drug development against multidrug efflux pumps

Abstract

Multidrug efflux pumps are important in the multidrug resistance of Gram-negative pathogens. However, despite efforts to develop efflux inhibitors, clinically useful inhibitors are not available at present. ABI-PP (a pryridopyrimidine derivative) is a MexB-specific inhibitor that does not inhibit MexY; MexB and MexY are principal pumps in Pseudomonas aeruginosa. We previously found that drugs were exported through tandem proximal and distal multisite drug-binding pockets. Here we describe the first inhibitor-bound structures of pumps. ABI-PP binds tightly to a narrow pit located in the distal pocket and sterically hinders the functional rotation. Phenylalanine is located at the edge of this pit in MexB and contributes to the tight binding of the inhibitor molecule. On the other hand, the voluminous side chain of tryptophan located at the corresponding position in MexY prevents inhibitor binding. For the development of universal inhibitors of MexB and MexY, it is important to avoid the steric hindrance of tryptophan in MexY. Now we are developing clinically useful inhibitors on the basis of the structural information obtained. Started from the ABI-PP structure, we designed many compounds that can bind to the inhibitor-binding pits of MexB and MexY. Some of designed compounds were actually synthesized and their inhibitory activity determined. Finally, we obtained some lead compounds that showed complete prevention of the growth of strains expressing MexB and MexY with low concentrations of antibiotics.