Towards automation of a valuable preclinical cardiac safety pharmacology assay: Evaluation of the effects of cardiac ion channel blockers on cardiac repolarisation in vitro

• Published in: Journal of pharmacological and toxicological methods 2007-09, Vol.56 (2), p.194-202
• Main Authors: Lightbown, Ian D., Johnson, Mark, Davis, Anne, Leaney, Joanne, Leishman, Derek J.
• Format: Article
• Language: English
• Subjects: (ICH) S7B guidelines; Action potential duration; Action Potentials - drug effects; Action Potentials - physiology; Animals; Anti-Arrhythmia Agents - pharmacology; Arrhythmias; Automation; Butylamines - pharmacology; Calcium Channel Blockers - pharmacology; Canine; Cardiac repolarisation; Cisapride - pharmacology; Dogs; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical - instrumentation; Drug Evaluation, Preclinical - methods; Electric Stimulation; Electrophysiologic Techniques, Cardiac - instrumentation; Electrophysiologic Techniques, Cardiac - methods; Female; Heart - drug effects; Heart - physiology; In Vitro Techniques; Ion Channels - antagonists & inhibitors; Ion Channels - physiology; Long QT Syndrome - diagnosis; Long QT Syndrome - physiopathology; Male; Microelectrodes; Phenethylamines - pharmacology; Potassium channels; Purkinje Fibers - drug effects; Purkinje Fibers - physiology; Purkinje fibre; QT interval; Reproducibility of Results; Safety pharmacology; Sulfonamides - pharmacology; Verapamil - pharmacology
• ISSN: 1056-8719; 1873-488X
• DOI: 10.1016/j.vascn.2007.03.008

Introduction: Purkinje fibre repolarisation assays are valuable tools for identifying compounds which affect cardiac ion channels. The throughput of compound testing in this assay is low therefore we designed a novel recording system to improve screening and animal tissue usage efficiencies. Methods: The system was used to evaluate compounds using standard sharp microelectrode techniques. Animal tissue usage efficiencies were quantified by adding up the total number of Purkinje fibres from which recordings were attempted and dividing this by the number of experimental data sets generated, to arrive at a ‘fibres per data set’ ratio. Test compounds were dofetilide (3 × 10 − 10 to 10 − 8  M), cisapride (10 − 8 to 3 × 10 − 7  M), terodiline (10 − 6 to 3 × 10 − 5  M) and verapamil (3 × 10 − 7 to 10 − 5  M). Results: Using the novel modified system, 21 data sets were generated from 29 fibres, compared to 24 data sets from 41 fibres using the conventional manual recording system, demonstrating a 24% improvement in the efficiency of animal tissue usage. Comparing data from the manual and modified systems revealed differences in absolute values for all parameters including APD 90 (308.73 ± 9.97 ms, n = 24, compared to 275.27 ± 8.25 ms, n = 21, respectively; P < 0.05). Differences in the magnitude of changes in action potential parameters between the systems were also evident for all compounds including terodiline (1 × 10 − 5  M) which caused a − 27.1 ± 16.5% reduction in APD 50 in the manual system, compared to a − 55.2 ± 2.2% reduction in the modified system. Discussion: Although the value of the present study is limited by the small sample sizes, it has demonstrated utility of the modified system in improving efficiency of animal tissue usage. It offers potential utility in a higher throughput screening environment for examining the electrophysiological properties of novel compounds in native cardiac tissues, particularly where functional patch clamp data are limited.