Cardiosafety Testing Based on CiPA at Scale with SyncroPatch 384 and Genedata Screener
1Sonja Stoelzle-Feix, 2 Ada X. Yee, 1Maria Giustina Rotordam, 2 Martin Ginkel, 1Nina Brinkwirth, 2 Gabriela Nass Kovacs,
1Nadine Becker, 2Guillaume Lemercier, 2 Stephan Heyse, 2 Stephan Steigele
1Nanion Technologies GmbH, Munich, Germany | sonja@nanion.de
2Genedata AG, Basel, Switzerland |screener@genedata.com

As part of the Comprehensive In Vitro Proarrhythmia Assay (CiPA) initiative, the FDA and other medical agencies have collaborated with pharma companies, academics, CROs,
and device companies to develop a more comprehensive, robust, mechanism-based in vitro assessment of arrhythmia risk for new drugs. The CiPA protocols consist of panelsof standardized in-vitro assays, incorporating defined tests against major cardiac ion channels, and thus enable large-scale, early discovery screens for liability prediction. Here
we present a tailored solution for upscaled and traceable cardiac safety studies performed according to the CiPA guidelines. A combination of automated patch clamp assays
run on Nanion SyncroPatch 384 instruments with automatic processing of the resulting data in the Genedata Screener software enables comprehensive cardiac risk
assessment on potentially thousands of compounds per week, following CiPA experimental and analysis guidelines. This integrated, efficient workflow enables collection ofhigh-quality electrophysiological data, their rigorous analysis, and their collation with non-APC assays (e.g. impedance data) for final panel assessment. We demonstrate thisusing hERG and Nav1.5 assays conducted in compliance with CiPA recommendations on experimental procedures. Data processing, use of control data, data normalization,
visualization of recording quality over time and other quality control metrics, assay end results and cross-assay safety scores are all combined in a result package that provides
a clear path for putative submission to regulatory authorities. The automation technology presented enables large-scale cardiac safety assessment of candidate molecules,
comprehensive in quality and dimensionality and authoritative by virtue of data integrity and accessibility, to eliminate risks earlier in the process of therapeutic development.

SyncroPatch 384 and Screener Integration

CiPA - the future of Cardiosafety testing

Four components

Comprehensive In Vitro Proarrhythmia Assay
Drug Effects on
Multiple Human
Cardiac Currents

In Silico Reconstruction of Human
Ventricular Cellular
Electrophysiology

In Vitro Effects on
Human Stem-Cell
Derived Ventricular
Myocytes

SyncroPatch 384, 384i, 768

Clinical Evaluation:
Unanticipated EPEffects

JSON• Raw channel recordings• Compound mapping• Operational parameters

High throughput APC

Automated analysis

CiPA Voltage Protocols
A current goal of CiPA is to determine requirements and protocols to deliverrobust, reliable and reproducible ion channel data in a high throughput screening(HTS) environment using high throughput automated patch clamp (APC) systems.

Nav1.5hERG

Reference Compounds Designated by the CiPA Initiative (“CiPA Compounds, May 10, 2016“)• Bepridil

Chlorpromazine•• Ondansetron •Quinidine •Cisapride•

Ranolazine ••Diltiazem

Sotalol ••Mexiletine

Terfenadine • Verapamil

Workflow for Cardiosafety Analysis
Normalized, Quality Results
per Sweep

Compound Results per Assay

Import Recordings &Metadata, Normalize

Individual well information and final compound results1 Compounds

Normalization to
CiPA-definedreferencecompounds andcontrol wells

Traces with cursors

2 Traces

3 Wells

Curves and peak amplitude
over time in thumbnail

Automated
quality control
according toCiPA
guidelines

Collation of the CiPA assays on multiple ionchannels and final safety assessment
Results from hERG
electrophysiology assay

Results from Nav1.5
electrophysiology assay

Results from
Impedance assay

Sweep & well information

3

1

Define criteria for automated QC

4Titration
Response

Make Decisions – And Share and Report Them4

2

Automated flaggingof compounds activein multiple assays

Create further summary visualizations(e.g. scatter plots)

Creation of detailed reports; export of recordings, finalresults, and assessment decisions to data warehouses…

Individual trace

Interactively mask ofoutlier replicates

For internal orexternal sharing

PNG

The direct interaction between the Nanion SyncroPatch 384 instrument and Genedata Screener enables seamless, rigorous analysis of hERG, Nav1.5, and other
CiPA-mandated ion channel assays in an automated workflow. It makes it easier to consistently apply CiPA guidelines to large-scale, early discovery screens andincorporate them as part of standard operating procedure, including full data traceability and reporting. Thus, it helps organizations pave the way towards scalable,
early safety assessment to increase discovery success and prepares them for a CiPA-mandated process including in silico models and exports into open, eCTDcompliant data formats currently in development by the FDA and industry.