Division of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug Administration, Silver Spring, MD, USA.
Division of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug Administration, Silver Spring, MD, USA; Vertex Pharmaceuticals (Europe) Ltd, Abingdon, Oxfordshire, UK.
J Pharmacol Toxicol Methods. 2022 Sep-Oct;117:107193. doi: 10.1016/j.vascn.2022.107193. Epub 2022 Jul 2.
According to the ICH S7B guideline, drug candidates are screened for hERG block prior to first-in-human testing to predict the likelihood of delayed repolarization associated with a rare, but life-threatening, ventricular tachyarrhythmia. The new ICH E14 Q&As guideline allows hERG results to be used in later clinical development for decision-making (Q&As 5.1 and 6.1). To pursue this path, the hERG assay should be conducted following the new ICH S7B Q&A 2.1 guideline, which calls for best practice considerations of the recording temperature, voltage protocol, stimulation frequency, recording/data quality, and concentration verification. This study investigated hERG block by cisapride, dofetilide, terfenadine, sotalol, and E-4031 - positive controls commonly used to demonstrate assay sensitivity - using the manual whole cell patch clamp method and an action potential-like voltage protocol presented at 0.2 Hz. Recordings were conducted at room and near physiological temperature. Drug concentrations were measured using samples collected during real patch clamp experiments and satellite experiments. Results showed temperature effects for E-4031, terfenadine, and sotalol, but not cisapride and dofetilide. Cisapride and terfenadine showed substantial concentration losses, largely due to nonspecific binding to the perfusion apparatus. Using concentrations measured from the real and satellite experiments to assess block potencies yielded comparable results, indicating that satellite sample collection may be viable for drugs with nonspecific binding concerns only. In summary, this study provides block potencies for 5 hERG positive controls, and serves as a case study for hERG assays conducted, and results illustrated in accordance with the new ICH E14/S7B Q&As.
根据 ICH S7B 指南,在进行首次人体测试之前,药物候选物会被筛选是否会阻断 hERG,以预测与罕见但危及生命的室性心动过速相关的延迟复极的可能性。新的 ICH E14 Q&A 指南允许将 hERG 结果用于后期临床开发中的决策(问答 5.1 和 6.1)。为了追求这条道路,hERG 测定应该遵循新的 ICH S7B Q&A 2.1 指南进行,该指南要求考虑记录温度、电压方案、刺激频率、记录/数据质量和浓度验证等最佳实践。本研究使用手动全细胞膜片钳法和 0.2 Hz 呈现的动作电位样电压方案,用顺式阿曲库铵、多非利特、特非那定、索他洛尔和 E-4031(通常用于证明测定敏感性的阳性对照药物)研究了 hERG 阻断作用。记录是在室温下和接近生理温度下进行的。使用在实际膜片钳实验和卫星实验过程中收集的样品来测量药物浓度。结果表明 E-4031、特非那定和索他洛尔的温度效应,但顺式阿曲库铵和多非利特则没有。顺式阿曲库铵和特非那定显示出明显的浓度损失,这主要是由于它们与灌注装置的非特异性结合。使用从实际和卫星实验中测量的浓度来评估阻断效力得出了可比的结果,这表明卫星样品采集对于具有非特异性结合问题的药物可能是可行的。总之,本研究提供了 5 种 hERG 阳性对照药物的阻断效力,并且是根据新的 ICH E14/S7B Q&A 进行 hERG 测定和结果说明的案例研究。