Article date: April 2006
By: Ari J Alexandrou, Rona S Duncan, Anneli Sullivan, Jules C Hancox, Derek J Leishman, Harry J Witchel, Joanne L Leaney in Volume 147, Issue 8, pages 905-916
The fluoroquinolone antibiotic moxifloxacin has been associated with the acquired long QT syndrome and is used as a positive control in the evaluation of the QT‐interval prolonging potential of new drugs. In common with other QT‐prolonging agents, moxifloxacin is known to inhibit the hERG potassium K+ channel, but at present there is little mechanistic information available on this action. This study was conducted in order to characterise the inhibition of hERG current (IhERG) by moxifloxacin, and to determine the role in drug binding of the S6 aromatic amino‐acid residues Tyr652 and Phe656.
hERG currents were studied using whole‐cell patch clamp (at room temperature and at 35–37°C) in an HEK293 cell line stably expressing hERG channels.
Moxifloxacin reversibly inhibited currents in a dose‐dependent manner. We investigated the effects of different voltage commands to elicit hERG currents on moxifloxacin potency. Using a ‘step‐ramp’ protocol, the IC50 was 65 μM at room temperature and 29 μM at 35°C. When a ventricular action potential waveform was used to elicit currents, the IC50 was 114 μM.
Block of hERG by moxifloxacin was found to be voltage‐dependent, occurred rapidly and was independent of stimulation frequency.
Mutagenesis of the S6 helix residue Phe656 to Ala failed to eliminate or reduce the moxifloxacin‐mediated block whereas mutation of Tyr652 to Ala reduced moxifloxacin block by ∼66%.
Our data demonstrate that moxifloxacin blocks the hERG channel with a preference for the activated channel state. The Tyr652 but not Phe656 S6 residue is involved in moxifloxacin block of hERG, concordant with an interaction in the channel inner cavity.
The fluoroquinolone antibiotic moxifloxacin has been associated with the acquired long QT syndrome and is used as a positive control in the evaluation of the QT‐interval prolonging potential of new drugs. In common with other QT‐prolonging agents, moxifloxacin is known to inhibit the hERG potassium K+ channel, but at present there is little mechanistic information available on this action. This study was conducted in order to characterise the inhibition of hERG current (IhERG) by moxifloxacin, and to determine the role in drug binding of the S6 aromatic amino‐acid residues Tyr652 and Phe656.
hERG currents were studied using whole‐cell patch clamp (at room temperature and at 35–37°C) in an HEK293 cell line stably expressing hERG channels.
Moxifloxacin reversibly inhibited currents in a dose‐dependent manner. We investigated the effects of different voltage commands to elicit hERG currents on moxifloxacin potency. Using a ‘step‐ramp’ protocol, the IC50 was 65 μM at room temperature and 29 μM at 35°C. When a ventricular action potential waveform was used to elicit currents, the IC50 was 114 μM.
Block of hERG by moxifloxacin was found to be voltage‐dependent, occurred rapidly and was independent of stimulation frequency.
Mutagenesis of the S6 helix residue Phe656 to Ala failed to eliminate or reduce the moxifloxacin‐mediated block whereas mutation of Tyr652 to Ala reduced moxifloxacin block by ∼66%.
Our data demonstrate that moxifloxacin blocks the hERG channel with a preference for the activated channel state. The Tyr652 but not Phe656 S6 residue is involved in moxifloxacin block of hERG, concordant with an interaction in the channel inner cavity.
British Journal of Pharmacology (2006) 147, 905–916. doi:10.1038/sj.bjp.0706678
DOI: 10.1038/sj.bjp.0706678
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