The isolated rabbit heart and Purkinje fibers as models for identifying proarrhythmic liability.
BACKGROUND
Delayed ventricular repolarization is associated with rare, but often fatal, polymorphic tachyarrhythmias named Torsades de Pointes. ICH S7B guideline recommends an integrated approach for cardiovascular preclinical evaluation of new drug candidates, including action potential assays (as a Purkinje fiber test) but also proarrhythmia models. The aim of this preliminary study was to compare the respective value of two preclinical in vitro rabbit cardiac preparations-the Purkinje fiber and the isolated perfused heart (Langendorff method)-based on effects of dofetilide, a selective IKr inhibitor.


METHODS
Transmembrane action potentials from rabbit Purkinje fibers were recorded using a conventional intracellular glass microelectrode. Electrocardiograms from rabbit isolated hearts were evaluated for QRS, QT and T wave durations (Tpeak-Tend). The pacing protocol was the same for both preparations (basal rate of 80 bpm and pacing of 40, 60 and 140 bpm). Dofetilide was tested in both systems at concentrations of 1, 3 and 10 nmol/L.


RESULTS
In Purkinje fibers dofetilide induced a concentration- and reverse use-dependent increase in action potential durations measured at 50 and 90% of repolarization. At 10 nmol/L, only 3/10 fibers showed early after depolarizations. In the isolated heart model, dofetilide also induced a similar concentration- and reverse use-dependent increase in QT-interval. From 3 nmol/L, major changes in T wave morphology, R-on-T extrasystoles and TdP were observed, mainly at low rate. Prior to arrhythmias, T wave shape and duration were markedly altered suggesting an increase in the heterogeneity of cardiac ventricular repolarization.


CONCLUSIONS
The effects of dofetilide were comparable in the two models for delayed repolarization but the isolated heart appears to be a better predictor for arrhythmias and a unique in vitro model to assess arrhythmogenic potential of QT prolonging compounds at least when associated with IKr/hERG inhibition.
Roche
Michel
M
Department of Drug Safety Evaluation, Safety Pharmacology Group, Sanofi-Aventis Recherche & Développement, 3 Digue d'Alfortville, 94140 Alfortville, France.
Renauleaud
Céline
C
Ballet
Véronique
V
Doubovetzky
Michel
M
Guillon
Jean-Michel
JM
D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000818:Animals; D001145:Arrhythmias, Cardiac / Q000188:drug therapy / Q000503:physiopathology*; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D006321:Heart / Q000187:drug effects / Q000502:physiology*; D066298:In Vitro Techniques; D008297:Male; D010627:Phenethylamines / Q000494:pharmacology / Q000627:therapeutic use; D011690:Purkinje Fibers / Q000187:drug effects / Q000502:physiology*; D011817:Rabbits; D013449:Sulfonamides / Q000494:pharmacology / Q000627:therapeutic use
D010627:Phenethylamines; D013449:Sulfonamides; C063533:dofetilide