Evaluation of a High-Throughput Fluorescence Assay Method for hERG Potassium Channel Inhibition

The number of projects in drug development that fail in late phases because of cardiac side effects such as QT prolongation can impede drug discovery and development of projects. The molecular target responsible for QT prolongation by a wide range of pharmaceutical agents is the myocardial hERG potassium channel. It is therefore desirable to screen for compound interactions with the hERG channel at an early stage of drug development. Here, the authors report a cell-based fluorescence assay using membrane potential-sensitive fluorescent dyes and stably transfected hERG channels from CHO cells. The assay allows semiautomated screening of compounds for hERG activity on 384-well plates and is sufficiently rapid for testing a large number of compounds. The assay is robust as indicated by a Z′ factor larger than 0.6. The throughput is in the range of 10,000 data points per day, which is significantly higher than any other method presently available for hERG. The data obtained with the fluorescence assay were in qualitative agreement with those from patch-clamp electrophysiological analysis. There were no false-positive hits, and the rate of false-negative compounds is currently 12% but might be further reduced by testing compounds at higher concentration. Quantitative differences between fluorescence and electrophysiological methods may be due to the use- or voltage-dependentactivity of the antagonists.

• Publication year

  2005

• Venue

  Journal of biomolecular screening

• Publication date

  2005-06-01

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1177/1087057104272045PMID 15964935
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• INTERNATIONAL CONFERENCE ON HARMONISATION OF TECHNICAL REQUIREMENTS FOR REGISTRATION OF PHARMACEUTICALS FOR HUMAN USE

  2006cited by this paper
• Patch clamping by numbers.

  2004cited by this paper
• Functional and pharmacological properties of canine ERG potassium channels.

  2003cited by this paper
• Troubleshooting problems with in vitro screening of drugs for QT interval prolongation using HERG K+ channels expressed in mammalian cell lines and Xenopus oocytes.

  2002cited by this paper
• High-Throughput Screening for Ion Channel Modulators

  2002cited by this paper
• A Novel Membrane Potential-Sensitive Fluorescent Dye Improves Cell-Based Assays for Ion Channels

  2002cited by this paper
• A High-Throughput HERG Potassium Channel Function Assay: An Old Assay with a New Look

  2002cited by this paper
• The Canine Purkinje Fiber: An In Vitro Model System for Acquired Long QT Syndrome and Drug-Induced Arrhythmogenesis

  2001cited by this paper
• Acute canine model for drug-induced Torsades de Pointes in drug safety evaluation-influences of anesthesia and validation with quinidine and astemizole.

  2001cited by this paper
• Molecular determinants of inactivation and dofetilide block in ether a-go-go (EAG) channels and EAG-related K(+) channels.

  2001cited by this paper
• Molecular biology of K(+) channels and their role in cardiac arrhythmias.

  2001cited by this paper
• Development and Evaluation of High Throughput Functional Assay Methods for hERG Potassium Channel

  2001cited by this paper
• Screening lead compounds for QT interval prolongation.

  2001cited by this paper
• Gastrointestinal prokinetic drugs have different affinity for the human cardiac human ether-à-gogo K(+) channel.

  2001cited by this paper
• Validation of FLIPR Membrane Potential Dye for High Throughput Screening of Potassium Channel Modulators

  2001cited by this paper
• [3H]dofetilide binding to HERG transfected membranes: a potential high throughput preclinical screen.

  2001cited by this paper
• QT-interval prolongation by non-cardiac drugs: lessons to be learned from recent experience

  2000cited by this paper
• High affinity blockade of the HERG cardiac K(+) channel by the neuroleptic pimozide.

  2000cited by this paper
• Effects of fluoroquinolones on HERG currents.

  2000cited by this paper
• Mechanism of block and identification of the verapamil binding domain to HERG potassium channels.

  1999cited by this paper
• A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays

  1999cited by this paper
• Effects of H1 Antihistamines on Animal Models of QTc Prolongation

  1999cited by this paper
• Block of HERG Potassium Channels by the Antihistamine Astemizole and its Metabolites Desmethylastemizole and Norastemizole

  1999cited by this paper
• Cell-based assays and instrumentation for screening ion-channel targets.

  1999cited by this paper
• Repolarizing K+ currents in rabbit heart Purkinje cells

  1998cited by this paper
• Blockage of the HERG human cardiac K+ channel by the gastrointestinal prokinetic agent cisapride.

  1997cited by this paper
• The inhibitory effect of the antipsychotic drug haloperidol on HERG potassium channels expressed in Xenopus oocytes

  1997cited by this paper
• A mechanism for the proarrhythmic effects of cisapride (Propulsid): high affinity blockade of the human cardiac potassium channel HERG

  1997influential reference
• Blockade of HERG channels expressed in Xenopus oocytes by the histamine receptor antagonists terfenadine and astemizole

  1996cited by this paper
• Multiple mechanisms in the long-QT syndrome. Current knowledge, gaps, and future directions. The SADS Foundation Task Force on LQTS.

  1996influential reference
• Ibutilide and the treatment of atrial arrhythmias. A new drug--almost unheralded--is now available to US physicians.

  1996cited by this paper
• Flow cytometric determination of absolute membrane potential of cells.

  1995cited by this paper
• A molecular basis for cardiac arrhythmia: HERG mutations cause long QT syndrome.

  1995cited by this paper
• Characterization of the steady-state and dynamic fluorescence properties of the potential-sensitive dye bis-(1,3-dibutylbarbituric acid)trimethine oxonol (Dibac4(3)) in model systems and cells.

  1994cited by this paper
• The Cardiac Arrhythmia Suppression Trial Investigators: Preliminary Report: Effect of Encainide and Flecainide on Mortality in a Randomized Trial of Arrhythmia Suppression After Myocardial Infarction.

  1990cited by this paper
• Preliminary report: effect of encainide and flecainide on mortality in a randomized trial of arrhythmia suppression after myocardial infarction.

  1989cited by this paper
• The long QT syndromes: a critical review, new clinical observations and a unifying hypothesis.

  1988cited by this paper
• ATP-Sensitive K+ Channels in Pancreatic β-Cells: Spare-Channel Hypothesis

  1988cited by this paper
• ATP-sensitive K+ channels in pancreatic beta-cells. Spare-channel hypothesis.

  1988cited by this paper
• Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches

  1981cited by this paper

• MMRCL: An interpretable multi-modal deep learning framework for predicting hERG blockers.

  2026cites this paper
• CardioGenAI: a machine learning-based framework for re-engineering drugs for reduced hERG liability

  2024cites this paper
• Utilizing Heteroatom Types and Numbers from Extensive Ligand Libraries to Develop Novel hERG Blocker QSAR Models Using Machine Learning-Based Classifiers

  2023cites this paper
• hERG-toxicity prediction using traditional machine learning and advanced deep learning techniques

  2023cites this paper
• Handbook of Ion Channels

  2023cites this paper
• hERG agonists pose challenges to web-based machine learning methods for prediction of drug-hERG channel interaction.

  2023cites this paper
• HergSPred: Accurate Classification of hERG Blockers/Nonblockers with Machine-Learning Models

  2022cites this paper
• Predicting hERG channel blockers with directed message passing neural networks

  2022cites this paper
• Virtual screening of DrugBank database for hERG blockers using topological Laplacian-assisted AI models

  2022cites this paper
• TSSF-hERG: A machine-learning-based hERG potassium channel-specific scoring function for chemical cardiotoxicity prediction.

  2021cites this paper
• Computational prediction of hERG blockers using homology modelling, molecular docking and QuaSAR studies

  2021cites this paper
• Drug discovery and development: An overview of modern methods and principles

  2021cites this paper
• Capsule Networks Showed Excellent Performance in the Classification of hERG Blockers/Nonblockers

  2020cites this paper
• hERG liability classification models using machine learning techniques

  2019cites this paper
• Intracellular Calcium Mobilization in Response to Ion Channel Regulators via a Calcium-Induced Calcium Release Mechanism

  2017cites this paper
• Natural products modulating the hERG channel: heartaches and hope† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c7np00014f

  2017cites this paper
• Modeling of the hERG K+ Channel Blockage Using Online Chemical Database and Modeling Environment (OCHEM)

  2017cites this paper
• Development of potent small-molecule inhibitors to drug the undruggable steroid receptor coactivator-3

  2016cites this paper
• Contribution of Automated Technologies to Ion Channel Drug Discovery.

  2016cites this paper
• Chapter 1 – Drug Discovery and Development: An Overview of Modern Methods and Principles

  2015cites this paper
• Cardiotoxicity screening: a review of rapid-throughput in vitro approaches

  2015cites this paper
• Novel intramolecular photoinduced electron transfer-based probe for the Human Ether-a-go-go-Related Gene (hERG) potassium channel.

  2015cites this paper
• Targeting cardiac potassium channels for state-of-the-art drug discovery

  2015cites this paper
• Fluorescence anisotropy (polarization): from drug screening to precision medicine

  2015cites this paper
• High-Throughput Methods for Ion Channels

  2015cites this paper
• Computer-based prediction of the drug proarrhythmic effect: problems, issues, known and suspected challenges.

  2014cites this paper
• In Vitro Methods for Cardiotoxicity Testing

  2014cites this paper
• Predicting Drug‐Induced QT Prolongation and Torsades de Pointes: A Review of Preclinical Endpoint Measures

  2013cites this paper
• New screening system for selective blockers of voltage-gated K(+) channels using recombinant cell lines dying upon single action potential.

  2013cites this paper
• hERG K(+) channels: structure, function, and clinical significance.

  2012cites this paper
• Development of Recombinant Cell Line Co-expressing Mutated Nav1.5, Kir2.1, and hERG for the Safety Assay of Drug Candidates

  2012cites this paper
• Antidepressants cause bradycardia and heart block in GD 13 rat embryos in vitro.

  2012cites this paper
• Role of amino acids in the lyophilization of RBC

  2012cites this paper
• The PADMET platform - pharmacokinetic characterization of carbohydrate mimetics

  2012cites this paper
• Preclinical Drug Safety and Cardiac Ion Channel Screening

  2011cites this paper
• Context Mediates Antimicrobial Efficacy of Kinocidin Congener Peptide RP-1

  2011cites this paper
• Preclinical Models for Anticancer Drug Development

  2011cites this paper
• Prospective Validation of a Comprehensive In silico hERG Model and its Applications to Commercial Compound and Drug Databases

  2010cites this paper
• Screening technologies for ion channel drug discovery.

  2010cites this paper
• Insights into the molecular mechanism of hERG1 channel activation and blockade by drugs.

  2010cites this paper
• Expanding the small molecular toolbox to study big biomolecular machines.

  2010cites this paper
• Iatrogenic QT Prolongation

  2010cites this paper
• Simulation Based Verifi cation of the Bayesian Approach in Single-Pool Hemodialysis Modelling

  2010cites this paper
• Selective reciprocity in antimicrobial activity versus cytotoxicity of hBD-2 and crotamine

  2009cites this paper
• Classifier Design to Improve Pattern Classification and Knowledge Discovery for Imbalanced Datasets

  2009cites this paper
• Should Cardiosafety be Ruled by hERG Inhibition? Early Testing Scenarios and Integrated Risk Assessment

  2009cites this paper
• Collation, assessment and analysis of literature in vitro data on hERG receptor blocking potency for subsequent modeling of drugs' cardiotoxic properties

  2009cites this paper
• In vitro profiling against ion channels beyond hERG as an early indicator of cardiac risk.

  2009cites this paper
• A new homogeneous high-throughput screening assay for profiling compound activity on the human ether-a-go-go-related gene channel.

  2009cites this paper
• Drug Binding to the Inactivated State Is Necessary but Not Sufficient for High-Affinity Binding to Human Ether-à-go-go-Related Gene Channels

  2008cites this paper
• Chapter 34 – hERG Methods

  2008cites this paper
• Recording hERG potassium currents and assessing the effects of compounds using the whole-cell patch-clamp technique.

  2008cites this paper
• Chapter 16 – hERG Blocking

  2008cites this paper
• Drug-like Properties: Concepts, Structure Design and Methods: from ADME to Toxicity Optimization

  2008cites this paper
• Prediction of hERG Potassium Channel Blockade Using kNN-QSAR and Local Lazy Regression Methods

  2008cites this paper
• The hERG potassium channel and hERG screening for drug-induced torsades de pointes.

  2008cites this paper
• Cell-Based Screening Assays

  2007cites this paper
• Early evaluation of compound QT prolongation effects: a predictive 384-well fluorescence polarization binding assay for measuring hERG blockade.

  2007cites this paper
• New cell models and assays in cardiac safety profiling

  2007cites this paper
• Optically recording the cardiac action potential from isolated ventricular myocytes

  2007influential citation
• New cell models and assays in cardiac safety profiling.

  2007cites this paper
• Fluorescently labeled analogues of dofetilide as high-affinity fluorescence polarization ligands for the human ether-a-go-go-related gene (hERG) channel.

  2007cites this paper
• Assessing the data quality in predictive toxicology using a panel of cell lines and cytotoxicity assays.

  2007cites this paper
• Predictive value of cellular methods in toxicological evaluation of lead candidates: an insight to hepatotoxicity.

  2007cites this paper
• A novel hypothesis for the binding mode of HERG channel blockers.

  2006cites this paper
• A Medicinal‐Chemistry‐Guided Approach to Selective and Druglike Sigma 1 Ligands

  2006cites this paper