Inhibition of aminoglycoside antibiotic resistance enzymes by protein kinase inhibitors.

Bacterial resistance to the aminoglycoside antibiotics is manifested primarily through the expression of enzymes which covalently modify these drugs. One important mechanism of aminoglycoside modification is through ATP-dependent O-phosphorylation, catalyzed by a family of aminoglycoside kinases. The structure of one of these kinases, APH(3')-IIIa has recently been determined by x-ray crystallography, and the general fold is strikingly similar to eukaryotic protein kinases (Hon, W. C., McKay, G. A., Thompson, P. R., Sweet, R. M., Yang, D. S. C., Wright, G. D., and Berghuis, A. M. (1997) Cell 89, 887-895). Based on this similarity, we have examined the effect of known inhibitors of eukaryotic protein kinases on two aminoglycoside kinases, APH(3')-IIIa and the enzyme AAC(6')-APH(2") which also exhibits acetyl-CoA-dependent aminoglycoside modification activity. We report that several known protein kinase inhibitors are also good inhibitors of aminoglycoside kinases. Compounds belonging to the isoquinolinesulfonamide group are especially effective in this regard, giving competitive inhibition in the micromolar range with respect to ATP and noncompetitive inhibition versus the aminoglycoside substrate. This study provides the basis for future aminoglycoside kinase inhibitor design and for the development of compounds which could reverse antibiotic resistance in the clinic.