A Place for High-Throughput Electrophysiology in Cardiac Safety: Screening hERG Cell Lines and Novel Compounds with the Ion Works HT™ System

Guthrie, Heather; Livingston, Frederick S.; Gubler, Ueli; Garippa, Ralph

doi:10.1177/1087057105280566

Cited by 32 publications

(15 citation statements)

References 18 publications

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“…PTA-6812) was generated and validated at Roche (Guthrie et al, 2005). Recombinant hERG K + channels originally cloned from human heart were stably expressed in CHO cells grown in sterile tissue flasks in DMEM/F-12 (1:1) medium (Invitrogen, USA) supplemented with 10% (v/v) heat-inactivated fetal calf serum (FCS) (Hyclone, USA) and 500 μg/ml gentamycin solution (Gibco, UK) at 35–37°C in 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Toward a New Gold Standard for Early Safety: Automated Temperature-Controlled hERG Test on the PatchLiner®

Polonchuk¹

2012

Front. Pharmacol.

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The Patchliner® temperature-controlled automated patch clamp system was evaluated for testing drug effects on potassium currents through human ether-à-go-go related gene (hERG) channels expressed in Chinese hamster ovary cells at 35–37°C. IC50 values for a set of reference drugs were compared with those obtained using the conventional voltage clamp technique. The results showed good correlation between the data obtained using automated and conventional electrophysiology. Based on these results, the Patchliner® represents an innovative automated electrophysiology platform for conducting the hERG assay that substantially increases throughput and has the advantage of operating at physiological temperature. It allows fast, accurate, and direct assessment of channel function to identify potential proarrhythmic side effects and sets a new standard in ion channel research for drug safety testing.

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Section: Methodsmentioning

confidence: 99%

Toward a New Gold Standard for Early Safety: Automated Temperature-Controlled hERG Test on the PatchLiner®

Polonchuk¹

2012

Front. Pharmacol.

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“…22 The assessment of compound activities on hERG channels is conducted with use of an automated, electrophysiological, ion flux method and so on. 7,23 The advantages of the hERG channel safety assay by use of IFM/Q3+Kir+hERG can be listed as follows: (1) The procedure and results are so simple that the assay including analyses can be performed without electrophysiological techniques. (2) The initial cost for equipment and running cost are low.…”

Section: Discussionmentioning

confidence: 99%

“…Accordingly, several methods in vitro are now available to evaluate the potential of drugs to suppress hERG or prolong the QT interval. The high-throughput screening (HTS) system for the hERG inhibition assay, including Rb + flux measurement, 5 potential sensitive fluorescence assay, 6 and improved automated patch clamp system, 7,8 have been also developed. New methods are, however, still emerging to evaluate the potential of test compounds to block the hERG K + channel as early as possible in the drug-development process with lower cost, smaller effort, and no exaggerated equipment.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2012

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To provide a high-throughput screening method for human ether-a-go-go-gene-related gene (hERG) K + channel inhibition, a new recombinant cell line, in which single action potential (AP)-induced cell death was produced by gene transfection. Mutated human cardiac Na + channel Nav1.5 (IFM/Q3), which shows extremely slow inactivation, and wild-type inward rectifier K + channel, Kir2.1, were stably co-expressed in HEK293 cells (IFM/Q3+Kir2.1). In IFM/Q3+Kir2.1, application of single electrical stimulation (ES) elicited a long AP lasting more than 30 s and led cells to die by more than 70%, whereas HEK293 co-transfected with wild-type Nav1.5 and Kir2.1 fully survived. The additional expression of hERG K + channels in IFM/Q3+Kir2.1 shortened the duration of evoked AP and thereby markedly reduced the cell death. The treatment of the cells with hERG channel inhibitors such as nifekalant, E-4031, cisapride, terfenadine, and verapamil, recovered the prolonged AP and dose-dependently facilitated cell death upon ES. The EC 50 values to induce the cell death were 3 µM, 19 nM, 17 nM, 74 nM, and 3 µM, respectively, whereas 10 µM nifedipine did not induce cell death. Results indicate the high utility of this cell system for hERG K + channel safety assay.

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“…Using CHO cells that stably express hERG channels, IonWorks allows for assessment of approximately 200 data points per Patch plate and about 80 concentration-response curves per day [21][22][23][24]. A good pharmacological correlation with conventional MPC data was observed in terms of rank-order of compounds with hERG activity although the concentration-response curves were shifted to the right (within 3-to 5-fold).…”

Section: Automated Planar Patch-clamp Methodsmentioning

confidence: 80%

Preclinical Drug Safety and Cardiac Ion Channel Screening

Gintant

2011

Heart Rate and Rhythm

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A new chemical entity must demonstrate efficacy and safety in order to be considered a successful therapeutic agent. Towards that goal, it is best to discover drugs demonstrating a wide concentration range between (lower) preclinical efficacious plasma concentrations and (higher) plasma concentrations eliciting preclinical off-target adverse effects. Drugs possessing such characteristics provide greater flexibility in clinical trials and explorations into alternative indications. Thus, preclinical studies related to both efficacy and safety must be considered equally important during drug discovery efforts. Cardiac safety plays a key role in defining the safety of novel therapeutics, and defining a drug's therapeutic window/safety margin.Much emphasis has been placed in the past decade on detecting (and avoiding) the ability of non-cardiovascular drugs to delay or alter ventricular repolarization (acquired long QT syndrome [1]). This focus arises in part from the association of delayed repolarization to the rare but potentially life-threatening arrhythmia torsades de pointes. As torsades is a rare event, surrogate markers are employed to detect and avoid this potential proarrhythmic risk. Preclinically, the most recognized in vitro marker for delayed repolarization is drug block of I Kr (an outward repolarizing current that initiates and defines terminal ventricular repolarization). In humans, the a-subunit of the I Kr channel is encoded by the human ether-a-go-go related gene (hERG) gene. Block of hERG/I Kr along with prolongation of the QT interval (an interval on the ECG that generally represents the onset of ventricular depolarization and the termination of ventricular repolarization) form the presentday cornerstone for cardiac electrophysiologic safety testing. Due to the importance of testing hERG current as a surrogate marker of proarrhythmia, great strides have been made in developing and streamlining testing protocols/procedures to evaluate

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A Place for High-Throughput Electrophysiology in Cardiac Safety: Screening hERG Cell Lines and Novel Compounds with the Ion Works HT™ System

Cited by 32 publications

References 18 publications

Toward a New Gold Standard for Early Safety: Automated Temperature-Controlled hERG Test on the PatchLiner®

Toward a New Gold Standard for Early Safety: Automated Temperature-Controlled hERG Test on the PatchLiner®

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

Preclinical Drug Safety and Cardiac Ion Channel Screening

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