The in vitro activity of ACHN-490, a novel aminoglycoside ("neoglycoside"), was evaluated against 102 multidrug-resistant (MDR) Klebsiella pneumoniae strains, including a subset of 25 strains producing the KPC carbapenemase. MIC 50 values for gentamicin, tobramycin, and amikacin were 8 g/ml, 32 g/ml, and 2 g/ml, respectively; MIC 90 values for the same antimicrobials were >64 g/ml, >64 g/ml, and 32 g/ml, respectively. ACHN-490 showed an MIC 50 of 0.5 g/ml and an MIC 90 of 1 g/ml, which are significantly lower than those of comparator aminoglycosides. ACHN-490 represents a promising aminoglycoside for the treatment of MDR K. pneumoniae isolates, including those producing KPC -lactamase.