Mechanistic Duality of Bacterial Efflux Substrates and Inhibitors: Example of Simple Substituted Cinnamoyl and Naphthyl Amides

D’Cunha, Napoleon; Moniruzzaman, Mohammad; Haynes, Keith; Malloci, Giuliano; Cooper, Connor J.; Margiotta, Enrico; Vargiu, Attilio V.; Uddin, Muhammad R.; Leus, Inga V.; Cao, Feng; Parks, Jerry M.; Rybenkov, Valentin V.; Ruggerone, Paolo; Zgurskaya, Helen I.; Walker, John K.

doi:10.1021/acsinfecdis.1c00100

Title: Mechanistic Duality of Bacterial Efflux Substrates and Inhibitors: Example of Simple Substituted Cinnamoyl and Naphthyl Amides

Journal Article · 11 August 2021 · ACS Infectious Diseases

DOI: https://doi.org/10.1021/acsinfecdis.1c00100 · OSTI ID:1820794

D’Cunha, Napoleon ^[1]; Moniruzzaman, Mohammad ^[2]; Haynes, Keith ^[1];

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Saint Louis University School of Medicine, St. Louis, MO (United States). Dept. of Pharmacology and Physiology
Univ. of Oklahoma, Norman, OK (United States). Dept. of Chemistry and Biochemistry
Univ. of Cagliari, Monserrato, Cagliari (Italy). Dept. of Physics
Univ. of Tennessee, Knoxville, TN (United States). Graduate School of Genome Science and Technology; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division
Dept. of Veteran Affairs Medical Center, St. Louis, MO (United States)

Antibiotic resistance poses an immediate and growing threat to human health. Multidrug efflux pumps are promising targets for overcoming antibiotic resistance with small-molecule therapeutics. Previously, we identified a diaminoquinoline acrylamide, NSC-33353, as a potent inhibitor of the AcrAB–TolC efflux pump in Escherichia coli. This inhibitor potentiates the antibacterial activities of novobiocin and erythromycin upon binding to the membrane fusion protein AcrA. It is also a substrate for efflux and lacks appreciable intrinsic antibacterial activity of its own in wild-type cells. Here, we have modified the substituents of the cinnamoyl group of NSC-33353, giving rise to analogs that retain the ability to inhibit efflux, lost the features of the efflux substrates, and gained antibacterial activity in wild-type cells. The replacement of the cinnamoyl group with naphthyl isosteres generated compounds that lack antibacterial activity but are both excellent efflux pump inhibitors and substrates. Surprisingly, these inhibitors potentiate the antibacterial activity of novobiocin but not erythromycin. Surface plasmon resonance experiments and molecular docking suggest that the replacement of the cinnamoyl group with naphthyl shifts the affinity of the compounds away from AcrA to the AcrB transporter, making them better efflux substrates and changing their mechanism of inhibition. These results provide new insights into the duality of efflux substrate/inhibitor features in chemical scaffolds that will facilitate the development of new efflux pump inhibitors.

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Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: National Institutes of Health (NIH); National Science Foundation (NSF); USDOE Office of Science (SC)

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1820794

Journal Information:: ACS Infectious Diseases, Journal Name: ACS Infectious Diseases Journal Issue: 9 Vol. 7; ISSN 2373-8227

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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Title: Mechanistic Duality of Bacterial Efflux Substrates and Inhibitors: Example of Simple Substituted Cinnamoyl and Naphthyl Amides

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