Multiple Ion Channel Effects Research Articles

Heart Rate Correction of the J-to-Tpeak Interval

Drug-induced changes of the J to T peak (JTp) and J to the median of area under the T wave (JT50) were reported to differentiate QT prolonging drugs that are predominant blockers of the delayed potassium rectifier current from those with multiple ion channel effects. Studies of drug-induced JTp/JT50 interval changes might therefore facilitate cardiac safety evaluation of new pharmaceuticals. It is not known whether formulas for QT heart rate correction are applicable to JTp and JT50 intervals. QT/RR, JTp/RR, and JT50/RR profiles were studied in 523 healthy subjects aged 33.5 ± 8.4 years (254 females). In individual subjects, 1,256 ± 220 electrocardiographic measurements of QT, JTp, and JT50 intervals were available including a 5-minute history of RR intervals preceding each measurement. Curvilinear, linear and log-linear regression models were used to characterize individual QT/RR, JTp/RR, and JT50/RR profiles both without and with correction for heart rate hysteresis. JTp/RR and JT50/RR hysteresis correction needs to be included but the generic universal correction for QT/RR hysteresis is also applicable to JTp/RR and JT50/RR profiles. Once this is incorporated, median regression coefficients of the investigated population suggest linear correction formulas JTpc = JTp + 0.150(1-RR) and JT50c = JT50 + 0.117(1-RR) where RR intervals of the underlying heart rate are hysteresis-corrected, and all measurements expressed in seconds. The established correction formulas can be proposed for future clinical pharmacology studies that show drug-induced heart rate changes of up to approximately 10 beats per minute.

Correlation between human ether-a-go-go-related gene channel inhibition and action potential prolongation.

Background and PurposeHuman ether‐a‐go‐go‐related gene (hERG; Kv11.1) channel inhibition is a widely accepted predictor of cardiac arrhythmia. hERG channel inhibition alone is often insufficient to predict pro‐arrhythmic drug effects. This study used a library of dofetilide derivatives to investigate the relationship between standard measures of hERG current block in an expression system and changes in action potential duration (APD) in human‐induced pluripotent stem cell‐derived cardiomyocytes (hiPSC‐CMs). The interference from accompanying block of Cav1.2 and Nav1.5 channels was investigated along with an in silico AP model.Experimental ApproachDrug‐induced changes in APD were assessed in hiPSC‐CMs using voltage‐sensitive dyes. The IC50 values for dofetilide and 13 derivatives on hERG current were estimated in an HEK293 expression system. The relative potency of each drug on APD was estimated by calculating the dose (D150) required to prolong the APD at 90% (APD90) repolarization by 50%.Key ResultsThe D150 in hiPSC‐CMs was linearly correlated with IC50 of hERG current. In silico simulations supported this finding. Three derivatives inhibited hERG without prolonging APD, and these compounds also inhibited Cav1.2 and/or Nav1.5 in a channel state‐dependent manner. Adding Cav1.2 and Nav1.2 block to the in silico model recapitulated the direction but not the extent of the APD change.Conclusions and ImplicationsPotency of hERG current inhibition correlates linearly with an index of APD in hiPSC‐CMs. The compounds that do not correlate have additional effects including concomitant block of Cav1.2 and/or Nav1.5 channels. In silico simulations of hiPSC‐CMs APs confirm the principle of the multiple ion channel effects.

CSAHi study: Detection of drug-induced ion channel/receptor responses, QT prolongation, and arrhythmia using multi-electrode arrays in combination with human induced pluripotent stem cell-derived cardiomyocytes

IntroductionThe use of multi-electrode arrays (MEA) in combination with human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) provides a promising method to predict comprehensive cardiotoxicity, including drug-induced QT prolongation and arrhythmia. We previously demonstrated that MEA in combination with hiPSC-CMs could provide a generalizable platform by using 7 reference drugs at 10 testing facilities. Using this approach, we evaluated responses to reference drugs that modulate a range of cardiac ion currents and have a range of arrhythmogenic effects. MethodsWe used the MEA system (MED64) and commercially available hiPSC-CMs (iCell cardiomyocytes) to evaluate drug effects on the beat rate, field potential duration (FPD), FPD corrected by Fridericia's formula (FPDc), and the incidence of arrhythmia-like waveforms. ResultsThis assay detected the repolarization effects of Bay K8644, mibefradil, NS1643, levcromakalim, and ouabain; and the chronotropic effects of isoproterenol, ZD7288, and BaCl2. Chronotropy was also affected by K+ and Ca2+ current modulation. This system detected repolarization delays and the arrhythmogenic effects of quinidine, cisapride, thioridazine, astemizole, bepridil, and pimozide more sensitively than the established guinea pig papillary muscle action potential assay. It also predicted clinical QT prolongation by drugs with multiple ion channel effects (fluoxetine, amiodarone, tolterodine, vanoxerine, alfuzosin, and ranolazine). DiscussionMEA in combination with hiPSC-CMs may provide a powerful method to detect various cardiac electrophysiological effects, QT prolongation, and arrhythmia during drug discovery. However, the data require careful interpretation to predict chronotropic effects and arrhythmogenic effects of candidate drugs with multiple ion channel effects.

Cardiotoxicity screening: a review of rapid-throughput in vitro approaches.

Cardiac toxicity represents one of the leading causes of drug failure along different stages of drug development. Multiple very successful pharmaceuticals had to be pulled from the market or labeled with strict usage warnings due to adverse cardiac effects. In order to protect clinical trial participants and patients, the International Conference on Harmonization published guidelines to recommend that all new drugs to be tested preclinically for hERG (Kv11.1) channel sensitivity before submitting for regulatory reviews. However, extensive studies have demonstrated that measurement of hERG activity has limitations due to the multiple molecular targets of drug compound through which it may mitigate or abolish a potential arrhythmia, and therefore, a model measuring multiple ion channel effects is likely to be more predictive. Several phenotypic rapid-throughput methods have been developed to predict the potential cardiac toxic compounds in the early stages of drug development using embryonic stem cells- or human induced pluripotent stem cell-derived cardiomyocytes. These rapid-throughput methods include microelectrode array-based field potential assay, impedance-based or Ca(2+) dynamics-based cardiomyocytes contractility assays. This review aims to discuss advantages and limitations of these phenotypic assays for cardiac toxicity assessment.

Quantitative Profiling of the Effects of Vanoxerine on Human Cardiac Ion Channels and its Application to Cardiac Risk.

Vanoxerine has been in clinical trials for Parkinsonism, depression and cocaine addiction but lacked efficacy. Although a potent blocker of hERG, it produced no serious adverse events. We attributed the unexpected result to offsetting Multiple Ion Channel Effects (MICE). Vanoxerine’s effects were strongly frequency-dependent and we repositioned it for treatment of atrial fibrillation and flutter. Vanoxerine terminated AF/AFL in an animal model and a dose-ranging clinical trial. Reversion to normal rhythm was associated with QT prolongation yet absent proarrhythmia markers for Torsade de Pointes (TdP). To understand the QT/TdP discordance, we used quantitative profiling and compared vanoxerine with dofetilide, a selective hERG-blocking torsadogen used for intractable AF, verapamil, a non-torsadogenic MICE comparator and bepridil, a torsadogenic MICE comparator. At clinically relevant concentrations, verapamil blocked hCav1.2 and hERG, as did vanoxerine and bepridil both of which also blocked hNav1.5. In acute experiments and simulations, dofetilide produced early after depolarizations (EADs) and arrhythmias, whereas verapamil, vanoxerine and bepridil produced no proarrhythmia markers. Of the MICE drugs only bepridil inhibited hERG trafficking following overnight exposure. The results are consistent with the emphasis on MICE of the CiPA assay. Additionally we propose that trafficking inhibition of hERG be added to CiPA.

GBR-12909: Potent Blocker of Peak and Late Nav1.5 Currents

GBR-12909 (vanoxerine) is a potent highly selective dopamine transporter antagonist. It was initially developed for treatment of Parkinson's disease and while safe in man, lacked efficacy. Although a potent hERG blocker, the drug exhibits multiple ion channel effects including frequency-dependence. The drug effectively terminated induced, sustained atrial fibrillation (AF) and atrial flutter in a canine model of sterile pericarditis. Subsequently the drug terminated AF in a Phase 2B ascending dose trial. Previous in vitro measurements used protocols that did not address the issue of the late Nav1.5 current which is important in understanding safety of a potent hERG blocker. Peak and late sodium currents were measured in HEK-293 cells stably expressing the human Nav1.5 channel. Peak currents were measured using a step-ramp protocol consisting of a 40 ms step to −15 mV (where the peak current was measured), a 180 ms step to +30 mV and a 200 ms ramp to the holding potential of −80 mV (1 Hz stimulating frequency). The effect of the drug was parameterized with a single binding isotherm having an IC50 of 29 ± 3 nM (95% CI: 22-37) and a Hill coefficient of 0.91 ± 0.10 (95% CI: 0.68-1.18) (seven cells, three concentrations). We also evaluated the effect of GBR-19209 on currents increased using the sea-anemone toxin ATX-II as a surrogate of the late sodium current. The calculated IC50 and Hill coefficients from five cells and five concentrations were 93 ± 9 nM (95% CI: 7-117) and 1.57 ± 0.24 (95% CI: 1.05-2.67), respectively. Thus, GBR-12909 block is at least sixty times more potent than ranolazine's (IC50 = 6 µM).The combination of potent peak and late sodium block may explain the antiarrhythmic and safe profile of the drug.

Effects of L-type calcium channel and human ether-a-go-go related gene blockers on the electrical activity of the human heart: a simulation study.

AimsClass III and IV drugs affect cardiac human ether-a-go-go related gene (IKr) and L-type calcium (ICaL) channels, resulting in complex alterations in repolarization with both anti- and pro-arrhythmic consequences. Interpretation of their effects on cellular and electrocardiogram (ECG)-based biomarkers for risk stratification is challenging. As pharmaceutical compounds often exhibit multiple ion channel effects, our goal is to investigate the simultaneous effect of ICaL and IKr block on human ventricular electrophysiology from ionic to ECG level.Methods and resultsSimulations are conducted using a human body torso bidomain model, which includes realistic representation of human membrane kinetics, anatomy, and fibre orientation. A simple block pore model is incorporated to simulate drug-induced ICaL and IKr blocks, for drug dose = 0, IC50, 2× IC50, 10× IC50, and 30× IC50. Drug effects on human ventricular activity are quantified for different degrees and combinations of ICaL and IKr blocks from the ionic to the body surface ECG level. Electrocardiogram simulations show that ICaL block results in shortening of the QT interval, ST elevation, and reduced T-wave amplitude, caused by reduction in action potential duration and action potential amplitude during the plateau phase, and in repolarization times. In contrast, IKr block results in QT prolongation and reduced T-wave amplitude. When ICaL and IKr blocks are combined, the degree of ICaL block strongly determines QT interval whereas the effect of IKr block is more pronounced on the T-wave amplitude.ConclusionOur simulation study provides new insights into the combined effect of ICaL and IKr blocks on human ventricular activity using a multiscale computational human torso model.

Automated Action Potential Recording in Stem Cell-Derived Cardiomyocytes for Proarrhythmia Assessment

Development of in vitro toxicity assays based on the human stem cell-derived cardiomyocytes (SC-CMs) provides an opportunity to assess the specific cardiac safety end-points early in drug development. The cardiac action potential (AP) is an established biomarker for cardiac safety and efficacy in humans and a good detector of proarrhythmic events. However, its utility has been mainly limited by a low throughput of manual patch clamp and a restricted access to healthy human cardiomyocytes. The recent progress in automated electrophysiology combined with availability of human SC-CMs provides an exciting possibility to measure AP in human cells expressing endogenous cardiac ion channels.Electrophysiological properties of human SC-CMs were investigated with special focus on establishment of automated cell-based cardiac safety screens.For the first time assay protocols and procedures were established to measure and characterize both cardiac AP and the main underlying currents in human SC-CMs using automated electrophysiology platforms Patchliner(r) and Synchropatch(r)96 (Nanion Technologies GmbH). Employment of the higher throughput systems allowed capturing and evaluating AP and cardiac current profiles in cardiomyocyte cultures revealing specific heterogeneity. Outward potassium current mainly consisting of slow and rapid delayed rectifier and Ca2+-activated components as well as inward cardiac sodium current were recorded and characterized. Density of L-type Ca2+ current varied with the type and differentiation status of cardiomyocytes. The inward rectifier potassium current and a pacemaker current could also be measured. These results demonstrate that functional cardiac currents and appropriate AP present in human SC-CMs can be successfully recorded by automated electrophysiology systems.The automated cardiac AP measurements in human SC-CMs exemplify a novel screening method for early and sensitive detection of multiple ion channel effects in vitro completing the current core battery hERG assay and animal ECG measurements in pre-clinical safety assessment.

MICE Models: Superior to the HERG Model in Predicting Torsade de Pointes

Drug-induced block of the cardiac hERG (human Ether-à-go-go-Related Gene) potassium channel delays cardiac repolarization and increases the risk of Torsade de Pointes (TdP), a potentially lethal arrhythmia. A positive hERG assay has been embraced by regulators as a non-clinical predictor of TdP despite a discordance of about 30%. To test whether assaying concomitant block of multiple ion channels (Multiple Ion Channel Effects or MICE) improves predictivity we measured the concentration-responses of hERG, Nav1.5 and Cav1.2 currents for 32 torsadogenic and 23 non-torsadogenic drugs from multiple classes. We used automated gigaseal patch clamp instruments to provide higher throughput along with accuracy and reproducibility. Logistic regression models using the MICE assay showed a significant reduction in false positives (Type 1 errors) and false negatives (Type 2 errors) when compared to the hERG assay. The best MICE model only required a comparison of the blocking potencies between hERG and Cav1.2.

Models based on multiple ion channel effects more predictive of cardiac risk

Models based on multiple ion channel effects more predictive of cardiac risk