CiPA - Comprehensive in vitro Proarrhythmia Assay
The Comprehensive In Vitro Proarrhythmia Assay (CiPA) initiative aims to replace the preclinical hERG current assay required under the ICH S7B safety pharmacology guidelines and clinical TQT study, which provides a surrogate marker of Proarrhythmia, with more translationally relevant assessments of proarrhythmic risk (Sager et al., 2014). CiPA intend to achieve this by evaluating proarrhythmic risk of evolving drug candidates based on an understanding of the electrophysiologic mechanisms responsible for proarrhythmia linked to Torsades de pointes (TdP) and QT prolongation.
The Q&As draft as endorsed August 2020 describe how nonclinical assays can be used as a part of an integrated risk assessment prior to first-in-human studies as they are used today, and in later stages of clinical development as a part of a combined nonclinical-clinical integrated risk assessment. If implemented, they can lead to a reduction in the number of ‘Thorough QT’ clinical studies and improved decision making at the time of a marketing application.
A webinar (Oct 15 and 16, 2020) will cover the background, motivation for, and overview of the new Q&As for ICH E14/S7B, followed by presentations on each of the main Q&A topics:
ICH E14 and S7B: Webinar on the recently released Draft Q&As
The CiPA workstreams
The Comprehensive in Vitro Proarrhythmia Assay (CiPA) paradigm was initiated in 2013. The Steering Team is comprised of partners from the US FDA, HESI, CSRC, SPS, Japan NIHS, PMDA, EMA and Health Canada. A number of participating organizations, amongst them Nanion Technologies, build the backbone of the initiative. The CiPA work streams include: In Silico, Myocyte, Ion Channel, and Clinical Translation working groups.
Nanion is a committee member of the Ion Channel HTS Team and a participant in the Myocyte Working Group. Nanion has a long-standing interest and extensive experience in automated patch clamp screening of cardiac ion channels. Label-free contractility and extracellular field potential recordings of stem cell-derived cardiomyocytes (iPSC-CMs) is also available in our portfolio. Our instruments are used for safety screening by major pharmaceutical companies and CROs worldwide and we are happy to assist you in setting up your CiPA assays.
Results of Phase I Studies
The Ion Channel Work Group finalized its phase I study in 2017. Besides further external sites, Nanion Technologies in Germany, USA and Japan participated with the Patchliner and the SyncroPatch 384PE in this study.
The image on the left hand side displays the results of the blocking effect of 12 compounds on hERG.
The results are in good agreement with manual patch clamp data (Crumb et al., 2016).
Nanion's products in CiPA workstreams
SyncroPatch 384i - Patch clamp meets HTS
The SyncroPatch 384i is a high throughput patch clamp instrument recording from up to 384 cells simultaneously. The SyncroPatch 384i is the highest throughput patch clamp instrument on the market with giga-seal data quality. The SyncroPatch 384/768PE and the new system SyncroPatch 384i are used for CiPA ion channel working group studies.
Request the CiPA assay protocol for the SyncroPatch 384i here.
Patchliner - Versatile and sophisticated
The Patchliner is a fully automated planar patch clamp instrument recording from up to 8 cells simultaneously. With its vast experimental freedom and gigaseal data quality, the Patchliner is one of the most versatile patch clamp instruments on the market.
The Patchliner is used for CiPA ion channel working group studies.
Request the CiPA assay protocol for the Patchliner here.
CardioExcyte 96 - Combined contractility, electrophysiology and cell viability
The CardioExcyte 96 is a hybrid system recording both contractility and electrophysiology of intact cardiomyocyte networks. In addition, the base impedance is continuously and automatically monitored as a measure of acute and chronic cell activity. No subtleties of cytotoxic responses are missed, this includes non-contractile cell types such as hepatocyte-like cells or cancer cells as well as contractile cardiac cells. Impedance and extracellular field potential measurements are performed at high resolution, are non-invasive and label-free. The CardioExcyte 96 is a fully automated device, recording from 96 wells at a time.
The CardioExcyte 96 is used for CiPA myocyte working group studies.
Request the CiPA assay protocol for the CardioExcyte 96 here.
Ion Channel Work Group (ICWG)
ICWG: Description and History
"The Ion Channel Working Group (ICWG), sponsored by the Safety Pharmacology Society (SPS), was established in December 2013. Its primary role is to work in close collaboration with the In Silico Working Group (ISWG) in providing ion channel support for the development of a computer model of the adult human ventricular myocyte, to be used as part of the CiPA initiative in predicting the clinical risk of drug-induced TdP."
"The original remit of the ICWG was: 1) to select key cardiac ion channels to include in the CiPA evaluation; 2) to develop robust, reliable and reproducible voltage clamp protocols required to generate data allowing the training and validation of the in silico model; 3) to define which biophysical and/or pharmacological properties of the channels to study for drug effects in order to optimize the predictivity of the model (e.g., potency (IC50), kinetic of block, rate/use/voltage dependence) and; 4) to define the requirements needed to transition the various ion channel protocols from manual to automated high throughput (HT) patch clamp platforms, in order to adapt to the screening environment present in most pharmaceutical companies."
Selected recombinant human channels: "IKr (hERG), ICa (L-type; Cav1.2), INa (Nav1.5 peak and late current); ITO (Kv4.3); IKs (KCNQ1 + KCNE1), and IK1 (Kir2.1). Following the selection, the ICWG set out to design standardized voltage clamp protocols for each of these channels."
(Source: Journal of Pharmacological and Toxicological Methods "The Comprehensive in Vitro Proarrhythmia Assay (CiPA) initiative — Update on progress. Colatzky et al. (2016) ")
Progress of the Ion Channel Work Group
"Protocols were developed and manual experimental work was completed. The High-Throughput Systems (HTS) studies to evaluate and characterize the automated patch clamp systems is underway. The training compounds have been completed for both manual and automated systems and data analysis initiated. The second phase of the HTS study with the validation compounds is underway. Additional ion channel studies will likely be planned beyond 2017 to further validate the model."
(Source: CiPAproject webpage "CiPA Workstream Timelines")
Data, Applications and Publications on the six Cardiac Ion Channels investigated by the "CiPA Ion Channel Work Group"
- NaV1.5 - Sodium Voltage-gated Channel Alpha Subunit 5
- CaV1.2 - voltage-dependent, L type, alpha 1C subunit calcium channel
- KV4.3 - Shal Related Potassium Channel Member 3
- KV7.1 - KVLQT1 - KQT Related Potassium Channel Member 1
- Kir2.1 - Potassium Voltage-Gated Channel Subfamily J Member 2
- hERG - KV11.1 - Erg Related Potassium Channel Member 2
Myocyte Work Group (MWG)
MWG: Description and History
The role of human cardiac stem cell derived cardiomyocytes (hSC-CMs) within the CiPA paradigm is to confirm in silico reconstructions of the electrophysiologic effects of drugs. This will be accomplished by evaluating the acute effects of drugs on the electrical activity of hSC-CM's using higher throughput approaches that enable more comprehensive and robust assessments earlier in the drug discovery process. Typical techniques involved include assessing changes in the extracellular field potentials of spontaneously active hSC-CMs using multielectrode array (MEA) platforms, or changes in the action potential configuration recorded optically using voltage-sensing dyes (VSD)."
"A pilot study was conducted by the Health and Environmental Sciences Institute (HESI) Cardiac Safety Committee Myocyte Subteam to evaluate reproducibility and variability of electrophysiologic responses across cells, platforms and volunteer study sites for 8 compounds, Mexiletine, Nifedipine, E-4031, JNJ-303, Flecanide, Moxifloxacin, Quinidine and Ranolazine. Overall, most sites detected changes in repolarization consistent with clinical findings, with the greatest source of variability attributed to study site."
"More recently, the HESI Myocyte Subteam was awarded a Broad Area Announcement Grant from the FDA to extend the pilot study to a larger set of 28 compounds categorized according to high, intermediate, and low/no risk of proarrhythmia based on clinical findings. The effects of these CiPA reference standards will be evaluated by a core group in this Phase II Validation study across four MEA and two VSO technology platforms using two commercially viable hSC-CM preparations." Results will be compared with prospective in silico reconstructions (based on voltage clamp studies) and published studies employing hSC-CMs from non-core laboratories and consortia for the same 28 compounds."
(Source: Journal of Pharmacological and Toxicological Methods "The Comprehensive in Vitro Proarrhythmia Assay (CiPA) initiative — Update on progress. Colatzky et al. (2016) ")
Progress of the Myocyte Work Group
The publication detailing the results of both the Pilot and Validation Studies are available online. The review on best practices is available here.
(Source: CiPAproject webpage "CiPA Workstream Timelines")
Nanion for CiPA:
Data and Applications
NaV1.5 - Late Current Analysis using the CiPA Protocol
SyncroPatch 384/768 PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing NaV1.5 current traces in response to the CiPA voltage step protocol, measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using whole cell patch clamp methodology and single-hole chips. The NaV1.5 late current was activated by the application of 60 nM ATX-II. The IC50 value of Ranolazine of the late sodium current current was determined as 40.4 µM.
hERG - Pharmacology at Physiological Temperature using the CiPA Protocol
SyncroPatch 384/768 PE (a predecessor model of the SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing hERG current traces in response to the CiPA voltage step protocol at 35 degree Celsius. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using perforated patch clamp methodology (Escin) and multi-hole chips (4 holes per well). The IC50 value of Erythromycin of the peak current was determined as 60.5 µM.
KV4.3 - Pharmacology of Metropolol Tartrate, using the CiPA Protocol
SyncroPatch 384/768 PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing KV4.3 current traces in response to the CiPA voltage step protocol, measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using the whole cell patch methodology and single-hole chips. The IC50 value of Metropolol Tartrate was determined as 128 µM.
CaV1.2 - Pharmacology of Nifedipine, using the CiPA protocol
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing hCaV1.2/β2/α2δ1 current traces in response to the CiPA voltage step protocol and the corresponing current-voltage relationship plot. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using perforated patch methodology (Escin) and multi-hole chips (4 holes per well), the success rate of valuable data for the analysis was 94%. The IC50 value of Nifedipine was determined as 106 nM.
hERG - Pharmacology of Cisapride, using the CiPA protocol
Patchliner data and applications:
Cells were kindly provided by Charles River.
The effect of cisapride on hERG currents was investigated, using the CiPA voltage step protocol. Measured on the Patchliner the perforated patch methodology (Escin) and multi-hole chips (4 holes per well) were used. The IC50 value of Cisapride was determined as 112 nM.
hERG - Pharmacology using the CiPA Protocol
SyncroPatch 384/768 PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing hERG current traces in response to the CiPA voltage step protocol. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using whole cell patch clamp methodology and multi-hole chips (4 holes per well). The IC50 value of the following compounds of the peak current was determined as 4.18 µM for Diltiazem, 37.4 nM for Terfenadine, 971 nM for Quinidine, 63 µM for Mexiletine, 431 nM for Verapamil and 4.54 µM for Ranolazine.
hERG - recordings with great stability using the CiPA step ramp protocol
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing hERG current traces in response to the CiPA voltage step protocol. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using perforated patch clamp methodology (Escin) and multi-hole chips (4 holes per well).
Cardiac Ion Channels - Pharmacology of Vandetanib
CardioExcyte 96 and Patchliner data and applications:
Cells were kindly provided by Charles River and Cellular Dynamics.
The image on the left hand side displays the results of the blocking effect of Vandetanib on hERG, NaV1.5, CaV1.2 and KV4.3. The compound induced arrhythmia when iPSC-CM were exposed to a minimum concentration of 1 µM. Arrhytmic events were both detected in field potential recordings as well as in the impedance based contractility.
Cardiac Ion Channels - Pharmacology of Sotalol
CardioExcyte 96 and SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River and Cellular Dynamics.
The image on the left hand side displays the results of the blocking effect of Sotalol on hERG. The result is in good agreement with manual patch clamp data (Crumb et al., 2016). The compound induced arrhythmia when iPSC-CM were exposed to a minimum concentration of 10 µM. Arrhytmic events were both detected in field potential recordings as well as in the impedance based contractility measurements.
Kir2.1 - Pharmacology of Barium
SyncroPatch 384/768 PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing Kir2.1 current traces in response to a voltage step protocol. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using the whole cell patch methodology and multi-hole chips (4 holes per well), the success rate of valuable data for the analysis was 93%. The IC50 value of Barium was determined as 6.38 µM (Literature: 16.2 µM, Schram et al. Cardiovasc Res. 2003).
KV4.3 - Pharmacology of Quinidine
SyncroPatch 384/768 PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing KV4.3 current traces in response to the CiPA voltage step protocol. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using the whole cell patch methodology and multi-hole chips (4 holes per well), the success rate of completed experiments was 95.3%. The IC50 value of Quinidine was determined as 21.2 µM (Literature: 79.3 µM, Crumb et al., J Pharmacol Toxicol Methods. 2016).
hERG - Application of "Sticky Compounds"
Patchliner data and applications:
hERG expressing HEK293 cells were kindly provided by Cytomyx/Millipore.
Even sticky compounds pose no problem for the Patchliner. IC50 measurements of well known sticky substances were determined on the Patchliner: Terfenadine IC50 = 11.0 ± 3 nM, Flunarizine IC50 163.7 ± 19 nM and Cisapride IC50 8.9 ± 3 nM.
NaV1.5 - Stable Access Resistance
Patchliner data and applications:
Cells were kindly provided by Millipore.
The I/V-characteristics of NaV1.5 currents (HEK293) are shown together with the repeated dose dependent block by TTX (lower panel). Five concentrations of TTX (0.3, 1, 3, 10, 30 μM) were applied, followed by washout with antagonist-free buffer and re-application of the same TTX concentrations.
hERG - Efficient Screening
Patchliner data and applications:
Cells were kindly provided by Cytomyx/Millipore.
The effects of six different blockers (terfenadine, cisapride, E4031, astemizole, propafenone, quinidine) on hERG currents (HEK293 cells) were investigated. Expected IC50 values for the different compounds were obtained. In two days, 119 full dose response curves were collected by a single person. Data was analyzed using Nanion’s Data Analysis Package, a very efficient and convenient data analysis tool!
hERG - Simple Data Analysis
Patchliner data and applications:
With our analysis tools, especially programmed routines in Igor make dose response curves, raw data and current time courses easily accessible. Also, creating average dose response curves over multiple experiments - even conducted on different days - remains easy.
hERG - Block at Physiological Temperature
Patchliner data and applications:
Cells were kindly provided by Cytomyx/Millipore, UK.
The effects of erythromycin on hERG currents were tested at different temperatures. Erythromycin has been shown to block hERG channels at physiological temperature with an IC50 of approx. 40 µM. However, at RT erythromycin is much less potent. For more details on these experiments please refer to the Application Note.
hERG - Stable Recordings with Accurate Pharmacology
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River Laboratories.
Current-voltage relationship of hERG (Kv11.1) expressed in HEK293 is shown along with pharmacology of 4 hERG-active compounds. The current-voltage relationships for all 384 wells (top) using perforated patch (Escin) and multi-hole chips (4 holes per well) are shown. In all 384 wells, a hERG-mediated current was observed with peak amplitude >700 pA at -20 mV. Using a pharmacology voltage protocol, experiments were stable lasting over 20 minutes. Concentration response curves for astemizole, pimozide, cisapride and terfenadine revealed IC50 values consistent with those found in the literature.
hERG - Stable Recordings
Patchliner data and applications:
Cells were kindly provided by Cytomyx/Millipore, UK.
A series of drug concentrations can be applied to each cell. The top figures show the original traces and the corresponding average dose-response curve. Five concentrations of Quinidine (0.1, 0.3, 1, 3 and 10 μM) have been applied.
The lower figure shows the corresponding Imax (-40 mV) including a wash out step and an additional application of the blocker to demonstrate the stability of whole cell recordings.
NaV1.5 - Lidocaine Block
Patchliner data and applications:
Cells were kindly provided by Cytomyx/Millipore.
Full dose response curves at different holding potentials were recorded for each cell (hNav1.5 in HEK293). Currents were elicited by a 10 ms voltage step to 0 mV. Plotted are average peak currents as a function of holding potential and lidocaine concentration.
Cardiomyocytes - Data from different cell providers
CardioExcyte data and applications:
Cells were kindly provided by FUJIFILM Cellular Dynamics, Nexel, Ncardia, Axiogenesis, Pluriomic, GE Healthcare, ReproCell, Takara Bio Cellartis Clonetech.
The CardioExcyte 96 allows for
• Non-invasive, label-free measurements of beating cardiomyocyte networks
• 96 recording wells in parallel with 1 ms time resolution
• Quick experiments or long-term compound effects on cardiotoxicity
• Real-time access to beating parameters
• Outstanding software for data analysis and export
• Cost effcient consumables - 96-well format
Cardiomyocytes - Tetracaine dose response curves as recorded with Cor.4U cells
CardioExcyte data and applications:
Cells were kindly provided by Axiogenesis.
Impedance amplitude is not changed by addition of increasing concentrations of Tetracaine (left panel), while beat rate of Cor.4U® cells is decreasing. For example, 29.6µM of Tetracaine decreased the beat rate by ~60% when compared to pre-addition values. Cumulative dose-response relationships indicate Tetracaine potency for same-well additions. Representative raw traces for impedance signals (middle panel) clearly indicate a decrease in cell monolayer beat rate with increasing concentrations of Tetracaine.
Extracellular Field Potential (EFP) spike amplitude is decreased by cumulative Tetracaine dose applications to the same monolayer of Cor.4U® cardiomyocytes (top right), in agreement with compound mechanism of action. Representative raw traces for EFP signals (bottom graph) clearly indicate a decrease in spike amplitude.
Cardiomyocytes - Nifedipine and its concentration dependent effect on Cor.4U cells
CardioExcyte data and applications:
Cells were kindly provided by Axiogenesis.
Nifedpinie is a dihydropyridine calcium channel blocker that primarily blocks L-type calcium channels. Impedance and EFP recordings on Cor.4U cells reveal a concentration dependent effect on impedance amplitude, beat rate and also a shortening of the FPD as expected.
Cardiomyocytes - Effect of E4031 on the impedance and EFP signals on iCell cardiomyocytes
CardioExcyte data and applications:
Cells were kindly provided by Cellular Dynamics.
Effect of the specific hERG blocker, E4031, on the impedance and EFP signals. A Impedance signal of 12 wells in control conditions (left) and the same 12 wells after 13 mins incubation in E4031 at the concentrations indicated (right). E4031 (300 nM, 3 mins incubation) induces EAD, shown in the inset, which can lead to potentially fatal ventricular arrhythmias. B EFP signal of 12 wells in control conditions (left) and the same 12 wells after 13 mins incubation in E4031 at the concentrations indicated (right). E4031(300 nM, 30 mins incubation) also causes arrhythmic effects in the EFP mode shown in the inset.
Cardiomyocytes - Effects of sotalol on impedance (left) and EFP (right) signals of Cellartis Cardiomyocytes
CardioExcyte data and applications:
Cells were kindly provided by Takara Bio Cellartis Clontech.
Impedance (left) and EFP signals (right) in control conditions (A) and 1 μM sotalol (B) are shown.
Irregular beating can be observed in the presence of sotalol.
Cardiomyocytes - Effects of verapamil on impedance (left) and EFP (right) signals of Cellartis Cardiomyocytes
CardioExcyte data and applications:
Cells were kindly provided by Takara Bio Cellartis Clontech.
(A) Impedance (left) and EFP signals (right) in control conditions
(B) Upon application of 10 nM verapamil
(C) Upon application of 100 nM verapamil.
(D) The mean beats for impedance (left) and EFP (right) are shown in D
Cardiomyocytes - Myocyte phase II study: CiPA conform analysis and arrhythmia detection
CardioExcyte data and applications:
Cells were kindly provided by Axiogenesis.
Nanion developed a CiPA conform analysis for the Myocyte phase II study. The feature comes along is included in our CiPA analysis routine. Automated arrhythmia detection is just one highlight out of many when it comes to the CardioExcyte 96 software.
CaV1.2 - Stable recording from frozen stock cells
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing hCaV1.2β2/α2δ1 current traces in response to a voltage step protocol and the corresponding current-voltage relationship plot. Measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384) using perforated patch methodology (Escin) and multi-hole chips (4 holes per well), the success rate of valuable data for the analysis was 100 %. The cells were used from a frozen cell stock (after induction) and recorded stably for more than 20 minutes. The IC50 value of Nifedipine was determined as 21 nM.
KV4.3/KChIP2 - Dose-response curve of Flecanaide
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing KV4.3/KChIP2 current traces in response to a voltage step protocol and the corresponding current-voltage relationship plot. Using whole cell mode in combination with multi-hole chips (4 holes per well), stably transfected cells were measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384). The IC50 value of flecainide was determined as 28.3 µM which is in accordance to literature. The success rate of completed experiments was 100%.
KV7.1 (KVLQT) - Dose-response curve
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing KV7.1/KCNE (KVLQT/minK) current traces in response to a voltage step protocol and the corresponding current-voltage relationship plot. Using the perforated patch methodology (Escin) in combination with multi-hole chips (4 holes per well), stably transfected cells were measured on the SyncroPatch 384PE (a predecessor model of SyncroPatch 384). The IC50 value of Chromanol 293B was determined as 3.82 µM. The success rate of completed exeriments was 100%.
hERG - Pharmacology of Bepridil, Dofetilide, Cisapride, Diltiazem (Results CiPA Phase I Study)
Patchliner data and applications:
Cells were kindly provided by Charles River.
The effect of four compounds on hERG currents were investigated, using the CiPA voltage step protocol. Measured on the Patchliner the perforated patch methodology (Escin) and multi-hole chips (4 holes per well) were used. The IC50 value of Cisapride was determined as 112 nM, Bepridil as 178 nM, Dofetilide as 33.9 nM and Diltiazem as 14.5 µM.
hERG - Pharmacology of Mexiletine, Quinidine, Ondansetron, Ranolazine (Results CiPA Phase I Study)
Patchliner data and applications:
Cells were kindly provided by Charles River.
The effect of four compounds on hERG currents were investigated, using the CiPA voltage step protocol. Measured on the Patchliner the perforated patch methodology (Escin) and multi-hole chips (4 holes per well) were used. The IC50 value of Mexiletine was determined as 77.3 µM, Quinidine as 1.04 µM, Ondansetron as 1.13 µM and Ranolazine as 11.9 µM.
hERG - Pharmacology of Sotalol, Terfenadine, Verapamil (Results CiPA Phase I Study)
Patchliner data and applications:
Cells were kindly provided by Charles River.
The effect of four compounds on hERG currents were investigated, using the CiPA voltage step protocol. Measured on the Patchliner the perforated patch methodology (Escin) and multi-hole chips (4 holes per well) were used. The IC50 value of Sotalol was determined as 157 µM, Terfenadine as 82.8 nM and Verapamil as 485 nM.
Application Notes
Cardiac Ion Channels - "High Throughput Screening of Cardiac Ion Channels"
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) Patchliner CardioExcyte 96 application note 
(2.3 MB)
Cardiac Ion Channels - "Simultaneous Assessment of CiPA Stipulated Ion Channels on the SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) application note (1.3 MB)
Cells were kindly provided by Charles River.
Cardiomyocytes - "Combining automated patch clamp, impedance and EFP of hiPSC-CMs"
CardioExcyte 96 SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) Patchliner Application Note
Cells kindly provided by Takara-Clonetech.
Cardiomyocytes - "Impedance and EFP recordings of Cor.4U cells using Nanion’s CardioExcyte 96"
CardioExcyte 96 Application Note (1.3 MB)
Cells were kindly provided by Ncardia.
Cardiomyocytes - "Impedance and EFP recordings of iCell Cardiomyocytes² on the CardioExcyte 96"
CardioExcyte 96 Application Note (2.8 MB)
Cells were kindly provided by Cellular Dynamics.
Cardiomyocytes - "Impedance and EFP recordings of Pluricyte Cardiomyocytes on the CardioExcyte 96"
CardioExcyte 96 Application Note (1.3 MB)
Cells were kindly provided by Ncardia.
CaV1.2 - "High Throughput Pharmacology of CaV1.2 Channels on Nanion’s SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) application note: (2.7 MB)
Cells were kindly provided by SB Drug Discovery.
CaV1.2 - "Stability and Pharmacology of CaV1.2 Channels on Nanion’s SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) application note (5.3 MB)
Cells were kidly provided by Charles River.
hERG - "Effect of temperature on erythromycin action on hERG currents recorded on Nanion's Patchliner"
Patchliner application note: (0.9 MB)
Cells were kindly provided by Millipore.
hERG - "High Throughput Pharmacology of hERG Channels on Nanion’s SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) application note (0.6 MB)
hiPSC-CM - "Conduction velocity with the 2-electrode layout of the CardioExcyte 96"
CardioExcyte 96 Application Note (1.4 MB)
Cardiosight®-S hiPSC-CMs kindly provided by Nexel
NaV1.5 - "Characterization of CreaCell's hNaV1.5 (A-0822) on Nanion's Patchliner"
Patchliner application note: (0.9 MB)
Cells were kindly provided by CreaCell.
NaV1.5 - "High Throughput Pharmacology of NaV1.5 Channels on Nanion's SyncroPatch 384PE"
SyncroPatch 384i (a predecessor model of the SyncroPatch 384) application note (1.5 MB)
Cells were kindly provided by Millipore.
NaV1.5 - "NaV1.5-ΔKPQ late INa current properties and pharmacology on the SyncroPatch 384i"
SyncroPatch 384i (a predecessor model of the SyncroPatch 384) application note (1.5 MB)
Cells were kindly provided by Metrion Biosciences.
NaV1.5 - "Pharmacology of hNaV1.5 recorded on Nanion's Patchliner"
Patchliner application note: (0.3 MB)
Cells were kindly provided by Millipore.
NaV1.5-Late - "INa-Late recorded from CHO cells and hiPSC-CMs on Nanion´s Patchliner"
Patchliner application note: (0.5 MB)
Cells were kindly provided by Charles River.
iCell® Cardiomyocytes2 were kindly provided by Fujifilm Cellular Dynamics International.
Publications
2021 - Applying the CiPA Approach to Evaluate Cardiac Proarrhythmia Risk of some Antimalarials Used Off‐label in the First Wave of COVID‐19
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384 instrument) prepublication in Clinical and Translational Science (2021)
Authors:
Delaunois A., Abernathy M., Anderson W.D., Beattie K.A., Chaudhary K.W., Coulot J., Gryshkova V., Hebeisen S., Holbrook M., Kramer J., Kuryshev Y., Leishman D., Lushbough I., Passini E., Redfern W.S., Rodriguez B., Rossman E.I., Trovato C., Wu C., Valentin J-P.
2020 - Time for a Fully Integrated Nonclinical–Clinical Risk Assessment to Streamline QT Prolongation Liability Determinations: A Pharma Industry Perspective
Highlighted Publication in Clinical Pharmacology & Therapeutics (2020)
Authors:
Vargas H.M., Rolf M.G., Wisialowski T.A., Achanzar W., Bahinski A., Bass A., Benson C.T., Chaudhary K.W., Couvreur N., Dota C., Engwall M.J., Foley C.M., Gallacher D., Greiter-Wilke A., Guillon J-M., Guth B., Himmel H.M., Hegele-Hartung C., Ito M., Jenkinson S., Chiba K., Lagrutta A., Levesque P., Martel E., Okai Y., Peri R., Pointon A., Qu Y., Teisman A, Traebert M., Yoshinaga T., Gintant G.A., Leishman D.J., Valentin J-P.
2020 - Repolarization studies using human stem cell-derived cardiomyocytes: Validation studies and best practice recommendations
CiPA review in Regulatory Toxicology and Pharmacology (2020)
Authors:
Gintant G., Kaushik E.P., Feaster T., Stoelzle-Feix S., Kanda Y., Osada T., Smith G., Czysz K., Kettenhofen R., Rong Lu H., Cai B., Shi H., Herron T.J., Dang Q., Burton F., Pang L., Traebert M., Abassi Y., Beck Pierson J., Blinova K.
2020 - Reliable identification of cardiac liability in drug discovery using automated patch clamp: Benchmarking best practices and calibration standards for improved proarrhythmic assessment
Patchliner and SyncroPatch 384i (SyncroPatch 384PEa predecessor model) publication in the Journal of Pharmacological and Toxicological Methods (2020)
Authors:
Brinkwirth N., Takasuna K., Masafumi D., Becker N., Obergrussberger A., Friis S., Furukawa Y., Hasegawa Y., Oka T., Ohtsuki A., Fertig N., Stoelzle-Feix S.
2020 - Integration of mechanical conditioning into a high throughput contractility assay for cardiac safety assessment
FLEXcyte 96 Publication in the Journal of Pharmacological and Toxicological Methods (2020)
Authors:
Gossmann M., Linder P., Thomas U., Juhasz K., Lemme M., George M., Fertig N., Dragicevic E., Stoelzle-Feix S.
2020 - Cross-site and cross-platform variability of automated patch clamp assessments of drug effects on human cardiac currents in recombinant cells
Patchliner and SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i instrument) publication in Nature Scientific Reports (2020)
Authors:
Kramer J., Himmel H.M., Lindqvist A., Stoelzle-Feix S., Chaudhary K.W., Li D., Bohme G.A., Bridgland-Taylor M., Hebeisen S., Fan J., Renganathan M., Imredy J., Humphries E.S.A, Brinkwirth N., Strassmaier T., Ohtsuki A., Danker T., Vanoye C., Polonchuk L., Fermini B., Pierson J.B. & Gintant G.
2020 - A systematic strategy for estimating hERG block potency and its implications in a new cardiac safety paradigm
Patchliner and SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i instrument) publication in Toxicology and Applied Pharmacology (2020)
Authors:
Ridder B.J., Leishman D.J., Bridgland-Taylor M., Samieegohar M., Han X, Wu W.W., Randolph A., Tran P., Sheng J., Danker T., Lindqvist A., Konrad D., Hebeisen S., Polonchuk L., Gissinger E., Renganathan M., Koci B., Wei H., Fan J., Levesque P., Kwagh J., Imredy J., Zhai J., Rogers M., Humphries E., Kirby R., Stoelzle-Feix S., Brinkwirth N., Rotordam M.G., Becker N., Friis S., Rapedius M., Goetze T.A., Strassmaier T., Okeyo G., Kramer J., Kuryshev Y., Wu C., Himmel H., Mirams G.R., Strauss D.G., Bardenet R., Li Z.
2020 - A general procedure to select calibration drugs for lab-specific validation and calibration of proarrhythmia risk prediction models: An illustrative example using the CiPA model
SyncroPatch 768PE (a predecessor model of the SyncroPatch 768i instrument) publication in the Journal of Pharmacological and Toxicological Methods (2020)
Authors:
Han X., Samieegohar M., Ridder B.J., Wu W.W., Randolph A., Tran P., Sheng J., Stoelzle-Feiz S., Brinkwirth N., Rotordam M.G., Becker N., Friis S., Rapedius M., Goteze T.A., Strassmaier T., Okeyo G., Kramer J., Kuryshev Y., Li Z.
2018 - Safety Pharmacology Methods and Models in an Evolving Regulatory Environment
CardioExcyte 96 & CiPA Editorial in Journal of Pharmacological and Toxicological Methods (2018)
Authors:
Pugsley M.K., Harter M.L., de Korte T., Connaughton C., Authier S., Curtis M.J.
2018 - Multifocal atrial and ventricular premature contractions with an increased risk of dilated cardiomyopathy caused by a Nav1.5 gain-of-function mutation (G213D)
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) -related publication in International Journal of Cardiology (2018)
Authors:
Calloe K., Broendberg A.K., Christensen A.H., Pedersen L.N., Olesen M.S., de los Angeles Tejada M., Friis S., Thomsen M.B., Bundgaard H., Jensen H.K.
2018 - Hypertrophic cardiomyopathy-linked mutation in troponin T causes myofibrillar disarray and pro-arrhythmic action potential changes in human iPSC cardiomyocytes
CardioExcyte 96 publication in Journal of Molecular and Cellular Cardiology
Authors:
Wang L., Kim K., Parikh S., Cadar A.G., Bersell K.R., He H., Pinto J.R., Kryshtal D.O., Knollmann B.C.
2018 - Dehydroevodiamine and hortiamine, alkaloids from the traditional Chinese herbal drug Evodia rutaecarpa, are IKr blockers with proarrhythmic effects in vitro and in vivo
Patchliner publication in Pharmacological Research (2018)
Authors:
Baburin I., Varkevisser R., Schramm A., Saxena P., Beyl S., Szkokan P., Linder T., Stary-Weinzinger A., van der Heyden M.A.G., Houtman M., Takanari H., Jonsson M., Beekman J.H.D., Hamburger M., Vos M.A., Hering S.
2018 - Cross-site comparison of excitation-contraction coupling using impedance and field potential recordings in hiPSC cardiomyocytes
CardioExcyte 96 publication in Journal of Pharmacological and Toxicological Methods (2018)
Authors:
Bot C.T., Juhasz K., Haeusermann F., Polonchuk L., Traebert M., Stölzle-Feix S.
2018 - A Hybrid Model for Safety Pharmacology on an Automated Patch Clamp Platform: Using Dynamic Clamp to Join iPSC-Derived Cardiomyocytes and Simulations of IK1 Ion Channels in Real-Time
Patchliner publication in Frontiers in Physiology
Authors:
Goversen B., Becker., N., Stölzle-Feix S., Obergrussberger A., Vos M.A., van Veen T.A.B., Fertig N., de Boer T.P.
2017 - L-Type Calcium Channel Inhibition Contributes to the Proarrhythmic Effects of Aconitine in Human Cardiomyocytes
Patchliner publication in PLoS ONE (2017)
Authors:
Wu J., Wang X., Chung Y.Y. Koh C.H. Liu Z., Guo H., Yuan Q., Wang C., Su S., Wei H.
2017 - Frequency-Dependent Multi-Well Cardiotoxicity Screening Enabled by Optogenetic Stimulation
CardioExcyte 96 publication in International Journal of Molecular Sciences
Authors:
Rehnelt S., Malan D., Juhasz K., Wolters B., Doerr L., Beckler M., Kettenhofen R., Bohlen H., Bruegmann T., Sasse P.
2017 - Correlation between human ether-a-go-go related gene channel inhibition and action potential prolongation
Patchliner publication in British Journal of Pharmacology (2017)
Authors:
Saxena P., Hortigon‐Vinagre M.P., Beyl S.,Baburin I., Andranovits S., Iqbal S.M., Costa A., IJzerman A.P., Kügler P., Timin E., Smith G.L., Hering S.
2017 - Combined Impedance and Extracellular Field Potential Recordings from Human Stem Cell-Derived Cardiomyocytes
CardioExcyte 96 book chapter in Stem Cell-Derived Models in Toxicology (2017)
Authors:
Obergrussberger A., Thomas U., Stölzle-Feix S., Becker N., Juhasz K, Doerr L., Beckler M., George M., Fertig N.
2017 - Automated Patch Clamp Recordings of Human Stem Cell- Derived Cardiomyocytes.
Patchliner and SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) book chapter in Stem Cell-Derived Models in Toxicology (2017)
Authors:
Obergrussberger A., Haarmann C., Stölzle-Feix S., Becker N., OhtsukiA., Brüggemann A., George M., Fertig N.
2017 - An impedance-based approach using human iPSC-derived cardiomyocytes significantly improves in vitro prediction of in vivo cardiotox liabilities
CardioExcyte 96 publication in Toxicology and Applied Pharmacology (2017)
Authors:
Koci B., Luerman G., Duenbostell A., Kettenhofen R., Bohlen H., Coyle L., Knight B., Ku W., Volberg W., Woska Jr. J.R., Brown M.P.
2016 - Use-dependent Block of Human Cardiac Sodium Channels by GS967
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) publication in Molecular Pharmacology (2016)
Authors:
Potet F., Vanoye C.G., George Jr. A.L.
2016 - Safety pharmacology studies using EFP and impedance
CardioExcyte 96 publication in Journal of Pharmacological and Toxicological Methods (2016)
Authors:
Obergrussberger A., Juhasz K., Thomas U., Stölzle-Feix S., Becker N., Dörr L., Beckler M., Bot C., George M., Fertig N.
2016 - Coupling Data Mining and Laboratory Experiments to Discover Drug Interactions Causing QT Prolongation
Patchliner publication in Journal of the American College of Cardiology (2016)
Authors:
Lorberbaum T., Sampson K.J., Chang J.B., Iyer V., Woosley R.L., Kass R.S., Tatonetti N.P.
2016 - Automated Patch Clamp Meets High-Throughput Screening: 384 Cells Recorded in Parallel on a Planar Patch Clamp Module
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) publication in Journal of Lab Automation (2016)
Authors:
Obergrussberger A., Brüggemann A., Goetze T.A., Rapedius M., Haarmann C., Rinke I., Becker N., Oka T., Ohtsuki A., Stengel T., Vogel M., Steindl J., Mueller M., Stiehler J., George M., Fertig N.
2016 - Automated Electrophysiological and Pharmacological Evaluation of Human Pluripotent Stem Cell-Derived Cardiomyocytes
Patchliner publication in Stem Cells and Development (2016)
Authors:
Rajamohan D., Kalra S., Hoang M.D., George V., Staniforth A., Russell H., Yang X., Denning C.
2015 - Novel screening techniques for ion channel targeting drugs
Patchliner, SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) and CardioExcyte 96 publication in Channels (2015)
Authors:
Obergrussberger A., Stölzle-Feix S., Becker N., Brüggemann A., Fertig N., Möller C.
2015 - New Easy-to-Use Hybrid System for Extracellular Potential and Impedance Recordings.
CardioExcyte 96 publication in Journal of Laboratory Automation (2015)
Authors:
Doerr L., Thomas U., Guinot D.R., Bot C.T., Stoelzle-Feix S., Beckler M., George M., Fertig N.
2015 - Electrophysiological analysis of mammalian cells expressing hERG using automated 384-well-patch-clamp
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) publication in BCM Pharmacology and Toxicology (2015)
Authors:
Haraguchi Y., Ohtsuki A., Oka T., Shimizu T.
2012 - Toward a new gold standard for early safety: automated temperature-controlled hERG test on the Patchliner
Patchliner publication in Frontiers in Pharmacology (2012)
Authors:
Polonchuk L.
2009 - High-throughput screening reveals a small-molecule inhibitor of the renal outer medullary potassium channel and Kir7.1
Patchliner publication in Molecular Pharmacology (2009)
Authors:
Lewis L.M., Bhave G., Chauder B.A., Banerjee S., Lornsen K.A., Redha R., Fallen K., Lindsley C.W., Weaver C.D., Denton J.S.
Posters
2020 - Unattended Screening Workflow in a 384-well Automated Patch Clamp System
SyncroPatch 384i (a predecessor model of SyncroPatch 384) poster, Safety Pharmacology Society Virtual Meeting 2020 (2.6 MB)
2020 - Reliable Identification of hERG Liability in Drug Discovery by Automated Patch Clamp
SyncroPatch 384i (a predecessor model of SyncroPatch 384) and Patchliner poster, 64th Annual Meeting of the Biophysical Society (1.3 MB)
2018 - High throughput automatic patch clamp: Applications for Safety Pharmacology
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384) poster, JSPS Meeting 2018 (2.3 MB)
2018 - Combining electrophysiology and contractility recordings for more complete assessment of hiPSC-CMs
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384), Patchliner and 
CardioExcyte 96 poster, Europhysiology Meeting 2018 (1.4 MB)
2018 - Assessment of Drug Effects on Cardiomyocyte Function: Comprehensive In Vitro Proarrhythmia Assay (CiPA) Results
CardioExcyte 96 poster, SOT Meeting 2018 (1.8 MB)
2015 - High Throughput Automated Patch Clamp of Ion Channels Important in Cardiac Safety and Drug Discovery
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) poster, Chantest Meeting 2015 (1.9 MB)
2015 - Complementary automated patch clamp, extracellular field potential and impedance recordings of iPSCs: safety screening tool box for the future
Patchliner and CardioExcyte 96 and SyncroPatch 384PE (a predecessor model of SyncroPatch 384) poster, SPS 2015 (2.7 MB)
Oral Presentations
2018 - HTS Phase I study: an update on progress of the CiPA Ion Channel Work Stream using the SyncroPatch 384PE and Patchliner
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i), Patchliner and CardioExcyte 96 Oral Presentation
Presenter:
Tim Strassmaier, Nanion Technologies Inc. USA
Source:
Webinar: "CiPA study: Bridging ion channel and myocyte data", September 12, 2018
2018 - CiPA myocyte phase II validation study results: cross-site comparison using the CardioExcyte 96
CardioExcyte 96 Oral Presentation
Presenter:
Dr. Sonja Stölzle-Feix, Nanion Technologies
Source:
Webinar: "CiPA study: Bridging ion channel and myocyte data", September 12, 2018
2017 - The CiPA initiative - Origin, Idea, Status, Issues, and Impact on Drug Development
Patchliner Oral Presentation
Presenter:
Dr. Herbert Himmel, Bayer Wuppertal, Germany
15.10.2020 | Webinar: ICH S7B best practices considerations - New Q&As and Benchmarking best practices
Patchliner and SyncroPatch 384i Webinar
Date: October 15. 2020
Speakers:
Dr. Sonja Stoelzle-Feix (Nanion Technologies)
This is an on-demand webinar from Nan]i[on and Friends 2020.
15.10.2020 | Webinar: Benchmarking best practices and calibration standards for HTS hERG recordings for improved proarrhythmic assessment
Patchliner and SyncroPatch 384i Webinar
Date: October 15. 2020
Speakers:
Dr. Alison Obergrussberger (Nanion Technologies)
This is an on-demand webinar from Nan]i[on and Friends 2020.
2020 - Validation of a impedance-based phenotypic screening assay able to detect multiple mechanisms of chronic cardiotoxicity in human stem cell-derived cardiomyocytes
CardioExcyte 96 presentation (slide deck) (4.5 MB)
2017 - HTS in Cardiac Safety
CardioExcyte 96 presentation (slide deck) (4.0 MB)
2017 - Cross-site comparison of myocyte phase II compounds on different iPS cardiomyocytes based on CardioExcyte 96 recordings
CardioExcyte 96 presentation (slide deck) (2.2 MB)
28.04.2015 | Webinar: Excited About Contraction – Combining Contractility and Excitability Measurements in Cardiotoxicity Screening
CardioExcyte 96
This Webinar covers the introduction of the CardioExcyte 96, a hybrid device for impedance- and MEA-type recordings from intact, beating networks of stem cell-derived cardiomyocytes.
15.10.2020 | Webinar: ICH S7B best practices considerations - New Q&As and Benchmarking best practices
Patchliner and SyncroPatch 384i Webinar
Date: October 15. 2020
Speakers:
Dr. Sonja Stoelzle-Feix (Nanion Technologies)
This is an on-demand webinar from Nan]i[on and Friends 2020.
15.10.2020 | Webinar: Benchmarking best practices and calibration standards for HTS hERG recordings for improved proarrhythmic assessment
Patchliner and SyncroPatch 384i Webinar
Date: October 15. 2020
Speakers:
Dr. Alison Obergrussberger (Nanion Technologies)
This is an on-demand webinar from Nan]i[on and Friends 2020.
12.09.2018 | Webinar: CiPA study: Bridging ion channel and myocyte data
CardioExcyte 96, Patchliner and SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) Webinar
Date: September 12, 4:00 PM CEST (10:00 AM EDT)
Get up-to-date with the CiPA progress of the Myocyte and Ion Channel Work Goups:
- CiPA myocyte phase II validation study results: cross-site comparison using the CardioExcyte 96
- HTS Phase I study: an update on progress of the CiPA Ion Channel Work Stream using the SyncroPatch 384PE and Patchliner
Instrument Product Sheets
FLEXcyte 96 product sheet 
CardioExcyte 96 - Product Sheet
CardioExcyte 96 product sheet (2.6 MB)
