ABSTRACT
To construct a non-clinical database for drug-induced QT interval prolongation, the electrophysiological effects of 11 positive and 10 negative compounds on action potentials (AP) in guinea-pig papillary muscles were investigated in a multi-site study according to a standard protocol. Compounds with a selective inhibitory effect on the rapidly activated delayed rectifier potassium current (IKr) prolonged action potential duration at 90% repolarization (APD90) in a concentration-dependent manner, those showing Ca2+ current (ICa) inhibition shortened APD30, and those showing Na+ current (INa) inhibition decreased action potential amplitude (APA) and Vmax. Some of the mixed ion-channel blockers showed a bell-shaped concentration-response curve for APD90, probably due to their blockade of INa and/or ICa, sometimes leading to a false-negative result in the assay. In contrast, all positive compounds except for terfenadine and all negative compounds with IKr-blocking activity prolonged APD30-90 regardless of their INa- and/or ICa-blocking activities, suggesting that APD30-90 is a useful parameter for evaluating the IKr-blocking activity of test compounds. Furthermore, the assay is highly informative regarding the modulation of cardiac ion channels by test compounds. Therefore, when APD90 and APD30-90 are both measured, the action potential assay can be considered a useful method for assessing the risk of QT interval prolongation in humans in non-clinical safety pharmacology studies.
Subject(s)
Action Potentials/drug effects , Biological Assay , Long QT Syndrome/chemically induced , Papillary Muscles/drug effects , Animals , Databases, Factual , Guinea Pigs , In Vitro Techniques , Male , Papillary Muscles/physiology , Pharmaceutical PreparationsABSTRACT
Certain compounds that prolong QT interval in humans have little or no effect on action-potential (AP) duration used traditionally, but they inhibit rapidly-activated-delayed-rectifier potassium currents (IKr) and/or human ether-a-go-go-related gene (hERG) currents. In this study using isolated guinea-pig papillary muscles, we investigated whether new parameters in AP assays can detect the inhibitory effects of various compounds on IKr and/or hERG currents with high sensitivity. The difference in AP duration between 60% and 30% repolarization, 90% and 60% repolarization, and 90% and 30% repolarization (APD30-60, APD60-90, and APD30-90, respectively) were calculated as the new parameters. All the 15 IKr and/or hERG current inhibitors that have been reported (9 compounds) or not reported (6 compounds) to inhibit calcium currents prolonged APD30-60, APD60-90, and/or APD30-90; and 8 of the 15 inhibitors prolonged APD30-60, APD60-90, and/or APD30-90 more potently than APD90. The APD30-60, APD60-90, and APD30-90 measurements revealed no difference in sensitivity when evaluating the effects of the IKr and/or hERG current inhibitors on the three parameters. On the other hand, compounds with little or no effect on hERG currents had no effect on APD30-60, APD60-90, or APD30-90. Therefore, it is concluded that in AP assays using isolated guinea-pig papillary muscles, APD30-60, APD60-90, and APD30-90 are useful indexes for evaluating the inhibitory effects of compounds including mixed ion-channel blockers on IKr and/or hERG currents.
Subject(s)
Action Potentials/drug effects , Calcium Channel Blockers/pharmacology , Long QT Syndrome/chemically induced , Papillary Muscles/drug effects , Potassium Channel Blockers/pharmacology , Animals , Databases, Factual , Delayed Rectifier Potassium Channels/antagonists & inhibitors , Delayed Rectifier Potassium Channels/physiology , Ether-A-Go-Go Potassium Channels/antagonists & inhibitors , Ether-A-Go-Go Potassium Channels/physiology , Guinea Pigs , In Vitro Techniques , Male , Papillary Muscles/physiologyABSTRACT
The aims of this study were to develop a suitable model to study proarrhythmic potential using isolated rabbit hearts with atrioventricular block and to examine the proarrhythmic potential of several drugs using this model. With a normal K/Mg solution (K+=5.7 mM and Mg2+=1 mM), d,l-sotalol (10 and 30 microM), a class III antiarrhythmic drug, prolonged ventricular repolarization, such as QT intervals and monophasic action potential duration, and induced early after-depolarization and polymorphic ventricular tachyarrhythmia. Cisapride (0.1 and 0.3 microM), a 5-HT4 receptor agonist, also prolonged the ventricular repolarization, and induced early after-depolarization. With a low K/Mg solution (K+=1.5 mM and Mg2+=0.35 mM), d,l-sotalol at 30 microM and cisapride at 0.3 microM more potently prolonged the ventricular repolarization than with a normal K/Mg solution. Furthermore, the incidence of polymorphic ventricular tachyarrhythmia caused by cisapride at 0.3 microM with a low K/Mg solution was higher than that with a normal K/Mg solution. Mosapride citrate, another 5-HT4 receptor agonist, at 10 microM prolonged the ventricular repolarization and induced early after-depolarization with a low K/Mg solution, whereas the drug at 1 and 3 microM did not affect any of the parameters examined. Des-4-fluorobenzyl-mosapride, a metabolite of mosapride citrate, at 10 microM slightly prolonged the ventricular repolarization without inducing early after-depolarization or ventricular tachyarrhythmia. These results suggest that mosapride citrate and des-4-fluorobenzyl-mosapride have much less proarrhythmic potential than cisapride and that isolated rabbit heart with atrioventricular block, perfused with a low K/Mg solution, is a suitable model for predicting the proarrhythmic potential of drugs.
