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How do large pharmaceutical companies mitigate concerns of drug-induced QT prolongation in drug discovery?
Abstracts
0032
AstraZeneca, Alderley Park, UK Johnson & Johnson, Beerse, Belgium c Abbvie, Abbott Park, IL, USA d Roche, Basel, Switzerland e Sanofi, Vitry Sur Seine, France f Bayer Pharma AG, Wuppertal, Germany g Astellas Pharma Inc., Tokyo, Japan h Eli Lilly & Co., Indianapolis, IN, USA i Bristol Myers Squibb, Princeton, NJ, USA j GlaxoSmithKline, Ware, UK k Pfizer, Groton, CT, USA l Merck & Co., West Point, PA, USA m Novartis Pharma AH, Basel, Switzerland n Amgen, Thousand Oaks, CA, USA o Takeda Pharmaceutical Company Limited, Osaka, Japan
OF
b
Several years after the implementation of ICH-S7A-B, we asked the question how do the top 15 pharmaceutical companies mitigate concerns of drug-induced QT prolongation in drug discovery?. Methods: A short anonymous survey was conducted, by invitation, to safety pharmacology representatives of the top 15 pharmaceutical companies defined by 2012 R&D (portfolio size). A series of multiple choice questions was designed to explore current practices in relation to utilization of in-silico, in-vitro and in-vivo approaches and deployment for hazard identification, risk assessment and management. Results: A 93% response rate was obtained. All responders strive to reduce “QT liability” during drug discovery by reducing absolute hERG potency alone (7%), increasing hERG safety margin alone (24%) or both (71%). Only 54% of responders use in-silico hERG models; these in-silico models are usually custom made or proprietary. Most responders (93%) do explore structure activity relationship to avoid hERG. During early drug discovery, most responders (79%) use additional in-vitro assays including i) ion channels other than hERG (91%), ii) other molecular targets (e.g. GPCRs, enzymes, transporters) (64%), iii) cell or tissue-based assays (82%), iv) whole organ based models (82%), and some others (including trafficking, in silico models, iPSC-derived cardiomyocytes or viable ex-vivo human organs or tissues) (27%). Finally, all responders try to reduce QT liability in-vivo by increasing the in-vivo QT safety margin. Conclusion: This survey highlighted similarity and differences in approaches employed by large pharmaceutical companies in mitigating QT prolongation while adding value to drug discovery.
TE
Predicting the pro-arrhythmic potential of compounds using human embryonic stem cell (hESC)-derived cardiomyocytes on multielectrode array Mike Clements, Jan Turner, Liz Roquemore
a
DP
doi:10.1016/j.vascn.2014.03.036
Chieko Kasaig, Derek Leishmanh, Paul Levesquei, James Louttitj, Sian Ratcliffek, Frederick Sannajustl, Willi Suterm, Hugo Vargasn, Keiji Yamamotoo
RO
decreases the potency of hERG blockers. This supports the hypothesis that block occurs via compound binding to the cytosolic-facing cavity of the channel. In contrast, the available data for hERG activators suggest an extracellular site of action. We tested this distinction by profiling hERG activator AZ1 (Bridgland-Taylor (2008), JPTM 58:156) in cell lines expressing WT, Y652A or F656T hERG. Concentration– effect (c–e) curves were generated using IonWorks to define IC50/ EC50 values for effects on pre-pulse current at + 40 mV and tail current at −30 mV (n = 4–20 cells per point on each c–e curve). To validate the hERG mutants, we first tested a hERG blocker (cisapride). As expected, there was a significant decrease in potency against hERG tail current relative to WT: 12-fold for Y652A; 9-fold for F656T. With Y652A, AZ1 blocked rather than enhanced both the pre-pulse and tail current (pre-pulse WT EC50 30.8 μM, Y652A IC50 14.8 μM; tail current WT EC50 12.8 μM, Y652A IC50 8.9 μM). In contrast, with the F656T mutant, its activating properties were enhanced (pre-pulse WT EC50 28.6 μM, F656T EC50 5.9 μM; tail current WT EC50 11.2 μM, F656T EC50 could not be determined owing to a “bell-shaped” curve but the ascending phase of the curve lies to the left of the WT data). These observations warrant further investigation but illustrate that AZ1's activator properties involve amino acids conventionally considered to be solely involved in channel block.
319
GE Healthcare, Cardiff, UK
UN
CO
RR
EC
Drug-induced delayed cardiac repolarization, a recognized risk factor for pro-arrhythmia, has become the single most common cause for the withdrawal of prescription drugs. The ability to identify detrimental off-target effects earlier has the potential to improve drug safety and reduce the cost of drug development. The vast majority of drugs known to prolong the repolarization of the cardiac membrane preferentially inhibit the delayed rectifier current (IKr) by binding to the hERG K+ channel. Consequently, functional in vitro assays for predicting a drug's potential to delay cardiac repolarization typically include evaluating hERG K+ channel block in transgenic cell lines, or action potential duration assays with primary canine or rabbit Purkinje fibers. The predictive value of these existing assays is limited, however, due to species differences and the lack of complex ion channel interactions in cell lines over-expressing hERG K+ channel. The introduction of assays utilizing human embryonic stem cell (hESC)-derived cardiomyocytes could potentially address the shortcomings of these existing models and form the basis of more predictive assays. Here, we describe the use of hESCderived cardiomyocytes on the multielectrode array platform (MEA) to assess the prospect of using the measured extracellular field potential as a pre-clinical cardiotoxicity screen. doi:10.1016/j.vascn.2014.03.037
doi:10.1016/j.vascn.2014.03.038
0034 Toward quasi-in vivo from in vitro assay (II): Importance of spatial arrangement of cardiomyocyte network for precise and stable in vitro drug screening measurement Fumimasa Nomura, Tomoyo Hamada, Hideyuki Terazono, Kenji Yasuda
0033
Tokyo Medical and Dental University, Tokyo, Japan
How do large pharmaceutical companies mitigate concerns of drug-induced QT prolongation in drug discovery? Jean-Pierre Valentina, David Bakera, David Gallacherb, Gary Gintantc, Andrea Greiter-Wilked, Jean-Michel Guillone, Herbert Himmelf,
We have investigated the community size and spatial arrangement dependence of cardiomyocyte network response against pharmaceutical compounds. Typical four types of cell networks of cardiomyocytes, small cluster, small closed loop, two-dimensional sheet, and a
0032
AstraZeneca, Alderley Park, UK Johnson & Johnson, Beerse, Belgium c Abbvie, Abbott Park, IL, USA d Roche, Basel, Switzerland e Sanofi, Vitry Sur Seine, France f Bayer Pharma AG, Wuppertal, Germany g Astellas Pharma Inc., Tokyo, Japan h Eli Lilly & Co., Indianapolis, IN, USA i Bristol Myers Squibb, Princeton, NJ, USA j GlaxoSmithKline, Ware, UK k Pfizer, Groton, CT, USA l Merck & Co., West Point, PA, USA m Novartis Pharma AH, Basel, Switzerland n Amgen, Thousand Oaks, CA, USA o Takeda Pharmaceutical Company Limited, Osaka, Japan
OF
b
Several years after the implementation of ICH-S7A-B, we asked the question how do the top 15 pharmaceutical companies mitigate concerns of drug-induced QT prolongation in drug discovery?. Methods: A short anonymous survey was conducted, by invitation, to safety pharmacology representatives of the top 15 pharmaceutical companies defined by 2012 R&D (portfolio size). A series of multiple choice questions was designed to explore current practices in relation to utilization of in-silico, in-vitro and in-vivo approaches and deployment for hazard identification, risk assessment and management. Results: A 93% response rate was obtained. All responders strive to reduce “QT liability” during drug discovery by reducing absolute hERG potency alone (7%), increasing hERG safety margin alone (24%) or both (71%). Only 54% of responders use in-silico hERG models; these in-silico models are usually custom made or proprietary. Most responders (93%) do explore structure activity relationship to avoid hERG. During early drug discovery, most responders (79%) use additional in-vitro assays including i) ion channels other than hERG (91%), ii) other molecular targets (e.g. GPCRs, enzymes, transporters) (64%), iii) cell or tissue-based assays (82%), iv) whole organ based models (82%), and some others (including trafficking, in silico models, iPSC-derived cardiomyocytes or viable ex-vivo human organs or tissues) (27%). Finally, all responders try to reduce QT liability in-vivo by increasing the in-vivo QT safety margin. Conclusion: This survey highlighted similarity and differences in approaches employed by large pharmaceutical companies in mitigating QT prolongation while adding value to drug discovery.
TE
Predicting the pro-arrhythmic potential of compounds using human embryonic stem cell (hESC)-derived cardiomyocytes on multielectrode array Mike Clements, Jan Turner, Liz Roquemore
a
DP
doi:10.1016/j.vascn.2014.03.036
Chieko Kasaig, Derek Leishmanh, Paul Levesquei, James Louttitj, Sian Ratcliffek, Frederick Sannajustl, Willi Suterm, Hugo Vargasn, Keiji Yamamotoo
RO
decreases the potency of hERG blockers. This supports the hypothesis that block occurs via compound binding to the cytosolic-facing cavity of the channel. In contrast, the available data for hERG activators suggest an extracellular site of action. We tested this distinction by profiling hERG activator AZ1 (Bridgland-Taylor (2008), JPTM 58:156) in cell lines expressing WT, Y652A or F656T hERG. Concentration– effect (c–e) curves were generated using IonWorks to define IC50/ EC50 values for effects on pre-pulse current at + 40 mV and tail current at −30 mV (n = 4–20 cells per point on each c–e curve). To validate the hERG mutants, we first tested a hERG blocker (cisapride). As expected, there was a significant decrease in potency against hERG tail current relative to WT: 12-fold for Y652A; 9-fold for F656T. With Y652A, AZ1 blocked rather than enhanced both the pre-pulse and tail current (pre-pulse WT EC50 30.8 μM, Y652A IC50 14.8 μM; tail current WT EC50 12.8 μM, Y652A IC50 8.9 μM). In contrast, with the F656T mutant, its activating properties were enhanced (pre-pulse WT EC50 28.6 μM, F656T EC50 5.9 μM; tail current WT EC50 11.2 μM, F656T EC50 could not be determined owing to a “bell-shaped” curve but the ascending phase of the curve lies to the left of the WT data). These observations warrant further investigation but illustrate that AZ1's activator properties involve amino acids conventionally considered to be solely involved in channel block.
319
GE Healthcare, Cardiff, UK
UN
CO
RR
EC
Drug-induced delayed cardiac repolarization, a recognized risk factor for pro-arrhythmia, has become the single most common cause for the withdrawal of prescription drugs. The ability to identify detrimental off-target effects earlier has the potential to improve drug safety and reduce the cost of drug development. The vast majority of drugs known to prolong the repolarization of the cardiac membrane preferentially inhibit the delayed rectifier current (IKr) by binding to the hERG K+ channel. Consequently, functional in vitro assays for predicting a drug's potential to delay cardiac repolarization typically include evaluating hERG K+ channel block in transgenic cell lines, or action potential duration assays with primary canine or rabbit Purkinje fibers. The predictive value of these existing assays is limited, however, due to species differences and the lack of complex ion channel interactions in cell lines over-expressing hERG K+ channel. The introduction of assays utilizing human embryonic stem cell (hESC)-derived cardiomyocytes could potentially address the shortcomings of these existing models and form the basis of more predictive assays. Here, we describe the use of hESCderived cardiomyocytes on the multielectrode array platform (MEA) to assess the prospect of using the measured extracellular field potential as a pre-clinical cardiotoxicity screen. doi:10.1016/j.vascn.2014.03.037
doi:10.1016/j.vascn.2014.03.038
0034 Toward quasi-in vivo from in vitro assay (II): Importance of spatial arrangement of cardiomyocyte network for precise and stable in vitro drug screening measurement Fumimasa Nomura, Tomoyo Hamada, Hideyuki Terazono, Kenji Yasuda
0033
Tokyo Medical and Dental University, Tokyo, Japan
How do large pharmaceutical companies mitigate concerns of drug-induced QT prolongation in drug discovery? Jean-Pierre Valentina, David Bakera, David Gallacherb, Gary Gintantc, Andrea Greiter-Wilked, Jean-Michel Guillone, Herbert Himmelf,
We have investigated the community size and spatial arrangement dependence of cardiomyocyte network response against pharmaceutical compounds. Typical four types of cell networks of cardiomyocytes, small cluster, small closed loop, two-dimensional sheet, and a