Received for publication July 5, 2007.
Revised September 13, 2007.
Accepted for publication September 14, 2007.

The molecular basis of high-affinity binding of the anti-arrhythmic compound, vernakalant (RSD1235), to Kv1.5 channels

Abstract

Vernakalant (RSD1235) is an investigational drug recently shown to convert atrial fibrillation rapidly and safely in patients (Roy et al., 2004). Here, the molecular mechanisms of interaction of vernakalant with the inner pore of the Kv1.5 channel are compared with the Class IC agent flecainide. Initial experiments showed that vernakalant blocks activated channels and vacates the inner vestibule as the channel closes, and thus mutations were made, targeting residues at the base of the selectivity filter and in S6, by drawing on studies of other Kv1.5-selective blocking agents. Block by vernakalant or flecainide of Kv1.5 wild-type (WT) and mutants was assessed by whole cell patch clamp experiments in transiently transfected HEK293 cells. The mutational scan identified several highly conserved amino acids, T479, T480, I502, V505 and V508, as important residues for affecting block by both compounds. In general, mutations in S6 increased the IC₅₀ for block by vernakalant, with I502A causing an extremely local 25-fold decrease in potency. Specific changes in the voltage-dependence of block with I502A supported the crucial role of this position. A homology model of the pore region of Kv1.5 predicted that, of these residues, only T479, T480, V505 and V508 are potentially accessible for direct interaction, and that mutation at additional sites studied may therefore affect block through allosteric mechanisms. For some of the mutations, the direction of changes in IC₅₀ were opposite for vernakalant and flecainide, highlighting differences in the forces that drive drug-channel interactions.

Key words: Antiarrhythmic drugs, Func. analysis receptor/ion channel mutants