title: Structure-guided optimization of protein kinase inhibitors reverses aminoglycoside antibiotic resistance.
abstract: Activity of the aminoglycoside phosphotransferase APH(3')-Ia leads to resistance to aminoglycoside antibiotics in pathogenic Gram-negative bacteria, and contributes to the clinical obsolescence of this class of antibiotics. One strategy to rescue compromised antibiotics such as aminoglycosides is targeting the enzymes that confer resistance with small molecules. We demonstrated previously that ePK (eukaryotic protein kinase) inhibitors could inhibit APH enzymes, owing to the structural similarity between these two enzyme families. However, limited structural information of enzyme-inhibitor complexes hindered interpretation of the results. In addition, cross-reactivity of compounds between APHs and ePKs represents an obstacle to their use as aminoglycoside adjuvants to rescue aminoglycoside antibiotic activity. In the present study, we structurally and functionally characterize inhibition of APH(3')-Ia by three diverse chemical scaffolds, anthrapyrazolone, 4-anilinoquinazoline and PP (pyrazolopyrimidine), and reveal distinctions in the binding mode of anthrapyrazolone and PP compounds to APH(3')-Ia compared with ePKs. Using this observation, we identify PP derivatives that select against ePKs, attenuate APH(3')-Ia activity and rescue aminoglycoside antibiotic activity against a resistant Escherichia coli strain. The structures described in the present paper and the inhibition studies provide an important opportunity for structure-based design of compounds to target aminoglycoside phosphotransferases for inhibition, potentially overcoming this form of antibiotic resistance.
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, Ontario, Canada.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Stogios
authors.forename: Peter
authors.initials: P
authors.lastname: Spanogiannopoulos
authors.forename: Elena
authors.initials: E
authors.lastname: Evdokimova
authors.forename: Olga
authors.initials: O
authors.lastname: Egorova
authors.forename: Tushar
authors.initials: T
authors.lastname: Shakya
authors.forename: Nick
authors.initials: N
authors.lastname: Todorovic
authors.forename: Alfredo
authors.initials: A
authors.lastname: Capretta
authors.forename: Gerard D
authors.initials: GD
authors.lastname: Wright
authors.forename: Alexei
authors.initials: A
authors.lastname: Savchenko
mesh_terms: D040981:Acinetobacter baumannii / Q000201:enzymology; D000617:Aminoglycosides / Q000494:pharmacology*; D000873:Anthracenes / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D015195:Drug Design; D024881:Drug Resistance, Bacterial / Q000187:drug effects*; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007612:Kanamycin / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D019868:Kanamycin Kinase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D008968:Molecular Conformation; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011720:Pyrazoles / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011743:Pyrimidines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011799:Quinazolines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D000617:Aminoglycosides; D000873:Anthracenes; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D007527:Isoenzymes; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011743:Pyrimidines; D011799:Quinazolines; D011994:Recombinant Proteins; D007612:Kanamycin; D019868:Kanamycin Kinase