hERG block potencies for 5 positive control drugs obtained per ICH E14/S7B Q&amp;As best practices: Impact of recording temperature and drug loss

Baron, Claudia Alvarez; Thiebaud, Nicolas; Ren, Ming; Viatchenko‐Karpinski, Serge; Indapurkar, Amruta; King, Tamara I.; Matta, Murali K.; Ismaiel, Omnia A.; Patel, Vikram; Mashaee, Manni; Vicente, José; Wu, Wendy

doi:10.1016/j.vascn.2022.107193

Cited by 7 publications

(6 citation statements)

References 24 publications

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“…Then, the superfusion of E‐4031 at 1 µM totally blocked the current. These effects are consistent with those previously described (Alexandrou et al., 2006; Alvarez Baron et al., 2022).…”

Section: Commentarysupporting

confidence: 93%

“…In order to limit premature drug discontinuation, several initiatives have been initiated, e.g., the Comprehensive In Vitro Proarrhythmia Assay (CiPA) paradigm and publication of the ICH guideline E14/S7B (ICH E14/S7B IWG, 2022). In addition, the variety of electrophysiological practices such as use of different medium compositions, stimulation protocols, and bath temperatures could lead to significant data variability and affect the results of the patch‐clamp assay (Alvarez Baron et al., 2022). The most recent ICH E14/S7B guidance (ICH E14/S7B IWG, 2022) is an important step in the evolutionary process of drug development, allowing harmonization of practices from one laboratory to another.…”

Section: Introductionmentioning

confidence: 99%

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Whole‐Cell Configuration of the Patch‐Clamp Technique in the hERG Channel Assay

Goineau,

Gallet,

Froget

2024

Current Protocols

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In vitro electrophysiological safety studies have become an integral part of the drug development process because, in many instances, compound‐induced QT prolongation has been associated with a direct block of human ether‐a‐go‐go‐related gene (hERG) potassium channels or their native current, the rapidly activating delayed rectifier potassium current (IKr). Therefore, according to the ICH S7B guideline, the in vitro hERG channel patch‐clamp assay is commonly used as an early screen to predict the ability of a compound to prolong the QT interval prior to first‐in‐human testing. The protocols described in this article are designed to assess the effects of acute or long‐term exposure to new chemical entities on the amplitude of IKr in HEK293 cells stably transfected with the hERG channel (whole‐cell configuration of the patch‐clamp technique). Examples of results obtained with moxifloxacin, terfenadine, arsenic, pentamidine, erythromycin, and sotalol are provided for illustrative purposes. © 2024 Wiley Periodicals LLC.Basic Protocol: Measurement of the acute effects of test items in the hERG channel testAlternate Protocol: Measurement of the long‐term effects of test items in the hERG channel test

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“…Then, the superfusion of E‐4031 at 1 µM totally blocked the current. These effects are consistent with those previously described (Alexandrou et al., 2006; Alvarez Baron et al., 2022).…”

Section: Commentarysupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Whole‐Cell Configuration of the Patch‐Clamp Technique in the hERG Channel Assay

Goineau,

Gallet,

Froget

2024

Current Protocols

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“…The main design component that many laboratories conducting the Good Laboratory Practice (GLP)‐compliant hERG assay will have to address is the incorporation of a concurrent positive control at 2 concentrations, above and below the anticipated IC 50 value. It is still common practice to use 1 high concentration of a positive control, often terfenadine or E4031, 42 thereby achieving 80% to 90% hERG inhibition. The new best practice will result in a more robust assessment of the IC 50 value for the positive control, which will then also make it easier to compare results with historical data, using the same experimental setting.…”

Section: Discussionmentioning

confidence: 99%

The New S7B/E14 Q&A Document Provides Additional Opportunities to Replace the Thorough QT Study

Darpö¹,

Leishman

2023

The Journal of Clinical Pharma

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Since 2015, concentration–QTc (C–QTc) analysis has been used to exclude the possibility that a drug has a concerning effect on the QTc interval. This has enabled the replacement of the designated thorough QT (TQT) study with serial electrocardiograms (ECGs) in routine clinical pharmacology studies, such as the first‐in‐human (FIH) study. The E14 revision has led to an increased proportion of FIH studies with the added objective of QT evaluation, with the intention of replacing the TQT study. With the more recent revision of the S7B/E14 Q&A document in February 2022, nonclinical assays/studies can be brought into the process of regulatory decisions at the time of marketing application. If the hERG (human ether‐a‐go‐go‐related gene) and the non‐rodent in vivo study are conducted according to the described best practices and are negative, the previous requirement that a QTc effect of >10 milliseconds must be excluded in healthy subjects at plasma concentrations 2‐fold above what can be seen in patients can be reduced to covering the concentrations seen in patients. For drugs that cannot be safely given in high doses to healthy subjects, ECG evaluation is often performed at the therapeutic dose in patients. If a QTc effect of >10 milliseconds can be excluded, an argument can be made that the drug should be considered as having a low likelihood of proarrhythmic effects due to delayedrepolarization, if supported by negative best practices hERG and in vivo studies. In this article, we describe what clinicians involved in early clinical development need to understand in terms of the hERG and in vivo studies to determine whether these meet best practices and therefore can be used in an integrated clinical/nonclinical QT/QTc risk assessment.

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“…Drug Response: To measure the impact of cell culture time on hiPSC-CM drug responsiveness, cells were exposed to three different drugs at two different time points (day 4, day 10 post-defrost; Figure 1A ). A hERG channel blocker (50 nM E-4031, #15203 Cayman Chemical) was tested for its ability to prolong FPD and APD(28, 42, 43). E-4031 reportedly suppresses spontaneous activity at concentrations above 30 nM (44).…”

Section: Methodsmentioning

confidence: 99%

hiPSC-CM Electrophysiology: Impact of Temporal Changes and Study Parameters on Experimental Reproducibility

Guerrelli,

Pressman,

Posnack

2023

Preprint

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Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are frequently used for preclinical cardiotoxicity testing and remain an important tool for confirming model-based predictions of drug effects in accordance with the Comprehensivein VitroProarrhythmia Assay (CiPA) initiative. Despite the considerable benefits hiPSC-CMs provide, concerns surrounding experimental reproducibility have emerged. Our study aimed to investigate the effects of temporal changes and experimental parameters on hiPSC-CM electrophysiology. hiPSC-CMs (iCell cardiomyocyte2) were cultured for 14 days and biosignals were acquired using a microelectrode array (MEA) system. Continuous recordings revealed a 22.6% increase in the beating rate and 7.7% decrease in the field potential duration (FPD) during a 20-minute equilibration period. Location specific differences across a multiwell plate were also observed, with hiPSC-CMs in the outer rows beating 8.8 beats per minute (BPM) faster than the inner rows. Cardiac endpoints were also impacted by cell culture duration; from 2-14 days the beating rate decreased (–12.7 BPM), FPD lengthened (+257 ms), and spike amplitude increased (+3.3 mV). Cell culture duration (4-10 days) also impacted hiPSC-CM drug responsiveness (E-4031, nifedipine, isoproterenol). Our study highlights multiple sources of variability that should be considered and addressed when performing hiPSC-CM MEA studies. To improve reproducibility and data interpretation, MEA-based studies should establish a standardized protocol and report key experimental conditions (e.g., culture time, equilibration time, electrical stimulation settings, report raw data values).New & NoteworthyWe demonstrate that hiPSC-CM electrophysiology measurements are significantly impacted by slight deviations in experimental techniques including electrical stimulation protocols, equilibration time, well-to-well variability, and length of hiPSC-CM culture. Furthermore, our results indicate that hiPSC-CM drug responsiveness changes within the first two weeks following defrost.Graphical AbstractImage created with Biorender.com

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hERG block potencies for 5 positive control drugs obtained per ICH E14/S7B Q&As best practices: Impact of recording temperature and drug loss

Cited by 7 publications

References 24 publications

Whole‐Cell Configuration of the Patch‐Clamp Technique in the hERG Channel Assay

Whole‐Cell Configuration of the Patch‐Clamp Technique in the hERG Channel Assay

The New S7B/E14 Q&A Document Provides Additional Opportunities to Replace the Thorough QT Study

hiPSC-CM Electrophysiology: Impact of Temporal Changes and Study Parameters on Experimental Reproducibility

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