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Assessment of the concordance between nonclinical repolarization assays and clinical measure of QT interval prolongation
Abstracts
Two novel approaches: Shifting the drug development paradigm Matthew S. Coffee, Javelyn R. Evans, David T. Merriman, Brian M. Roche
179
A retrospective analysis of gastrointestinal adverse events impacting on drug development Jean-Pierre Valentin, A. Al-Saffar, L. Ewart, J. Glab, L. Marks, W.S. Redfern, S. Roberts, T.G. Hammond
Battelle, Columbus, OH, United States AstraZeneca, Safety Assessment, Macclesfield, United Kingdom Over the last 10 years, there has been a significant decline in the number of new drugs being submitted to regulators, while over the same period, the number of regulatory approvals has also declined. This is seen as a result of the ever-increasing cost of drug development. A new drug-development model needs to emerge in order for drug-discovery programs to remain viable into the future. This new model's priorities must include better nonclinical testing, more collaborations or partnerships, and earlier involvement from the research and development provider during the drug discovery phase. This will allow the pharmaceutical industry to make more informed decisions regarding safety and efficacy sooner in the drugdevelopment process. Two approaches to meet this need are being evaluated to develop biobetters and to repurpose failed biologic drugs. Both approaches will ultimately increase the number of medicines available to consumers at a lower cost, thereby shifting the drug-development paradigm in an attempt to revitalize the drug industry. doi:10.1016/j.vascn.2012.08.070
The development of an in vitro safety assay utilising human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) with Multi/micro-electrode array (MEA) technology Amy M. Nicks, Yi Cui, Nick McMahon, Arun Sridhar
We conducted a literature and AZ portfolio review to assess the incidence, prevalence and impact of gastrointestinal (GI) adverse events (AEs) on drug development. In the last few decades, only a small proportion of drugs have been discontinued from development or withdrawn from the market due to GI-AEs (2–6%). However, data from the 2010 ‘DIA Daily Alert’ showed that GI-AEs accounted for 11% of delayed or halted clinical trials; an increase from previous years and a level identical to hepatotoxicity. Furthermore, GI-AEs featured significantly in serious Adverse Drug Reactions (ADRs; 14–23%) and label ADRs (67%). A PharmaPendiumTM search showed that 13% of clinical observations reported were GI disorders, the highest ranking for all organ systems. AEs, reported with 104 marketed drugs in Japan, showed a high incidence of GI-AEs (nausea and vomiting N diarrhoea N anorexia N stomach ache N constipation). Some plausible associations with preclinical GI findings were reported (e.g. intestinal transport, gastric emptying time, orgastric juice secretion vs. anorexia). An AstraZeneca portfolio review of the Phase I single ascending dose clinical trial AE profiles showed a high incidence of high stringency GI-AEs (38% of 95 drugs); an evaluation of the predictive value of nonclinical models is on-going. Ultimately, GI-AEs can be dose limiting preclinically and/or clinically. This affects patient compliance, and can alter the PK profile of drugs. This review suggests that the evaluation of GI liability should be carefully considered prior to human administration.
doi:10.1016/j.vascn.2012.08.072 GlaxoSmithKline, Ware, United Kingdom Adverse cardiac electrophysiological effects are common causes of preclinical drug failure. Multi-micro electrode array (MEA) technology with human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) has the potential to offer a high throughput, more humanlike predictive cardiac electrophysiological model. These experiments identify some of the optimal conditions for this novel assay. The hiPSCCMs were plated at 8000 cells/μl in 10 μl onto MEAs and the extracellular field potential of the electrically connected monolayer that beats spontaneously was recorded. The field potential duration (FPD) and beating rate of hiPSC-CMs were measured from an environmental chamber and compared to a dry incubator. These parameters were also measured in the presence of 0.1% DMSO or E4031 in the dry incubator. The rate of hiPSC-CMs was higher and more variable in the environmental chamber than in the dry incubator. 0.1% DMSO (n = 4) decreased rate by 8%, increased FPD by 6% and FPDc by 8% from baseline values of 0.6 Hz, 761 ± 31 ms and 895 ± 37 ms, respectively, these were not statistically significant (p N 0.05). Preliminary results showed that E-4031 prolonged the FPD at 30 nM and 100 nM by 23% and 34% respectively. To achieve a stable beating rate over time a dry incubator was used; this highlighted the sensitivity of hiPSC-CMs to temperature and CO2. Time dependent changes were seen in 0.1% DMSO; however, they were not significant. E-4031 showed concentration dependent changes confirming its effect of QT prolongation in the clinic (Okada Y et al, 1996). MEA technology and hiPSC-CMs demand further pharmacological validation to define their suitability for predicting clinical effects. doi:10.1016/j.vascn.2012.08.071
Assessment of the concordance between nonclinical repolarization assays and clinical measure of QT interval prolongation John Koernera, B. Darpob, M. Skinnerc, H.M. Vargasd, J. Willarda, S. Pettite, J.P. Valentinc a
U.S. FDA, Silver Spring, MD, United States Pharmaceutical Consultant, Sweden c AstraZeneca, Macclesfield, United Kingdom d Amgen, Inc, Thousand Oaks, CA, United States e HESI on behalf of the Pro-Arrhythmia Model Committee, Washington, DC, United States b
Drug-induced QT interval prolongation and Torsades de Pointes (TdP) remain serious public health issues in bringing safe new pharmaceuticals to the market place. Under the auspices of ILSI Health and Environmental Sciences Institute (HESI), a consortium including representatives from pharmaceutical companies, regulatory agencies and opinion leaders from the scientific and medical research communities has been initiated. The objectives are: to assess the concordance between nonclinical repolarization assays and clinical measures of QT interval prolongation; establish the sensitivity, specificity and predictivity of nonclinical assays towards the clinical outcome; and ultimately to identify compounds that do not show evidence of concordance. Four approaches are pursued and will be summarised: 1. Analysis of the concordance of data extracted from thorough clinical QT studies and nonclinical studies (i.e., in vitro hERG assay, action potential duration in isolated cardiac tissue, and in vivo QTc studies) submitted to the U.S. Food and Drug Administration to support marketing approval of new pharmaceuticals (N 70 drugs in the database). 2. Attempts to include Phase I clinical QTc data to the above analysis.
180
Abstracts
3. Review of the literature using bioinformatics and text mining tools (N120 drugs considered). 4. Retrospective quantitative analysis of the 2001 ILSI-HESI sponsored programme of work (Hanson et al., 2006) involving 12 drugs with clear or no associations with QTc prolongation and TdP. doi:10.1016/j.vascn.2012.08.073
Which possible applications for bioluminescence and fluorescence in preclinical safety studies? Anne Maurin, Eric Martel, Emmanuel Bracq, Laura Brulle, Pascal Champeroux, Serge Richard CERB, Baugy, France Bioluminescence and fluorescence are more and more used to assess the pharmacological activities of NCE in vivo. Their main advantages are they are not invasive, harmlessness and allow establishing kinetics on the same animal as illustrated with our results of orthotopic, colorectal, pancreatic and bones cancers in mice. In preclinical development, Fluorescence and Bioluminescence Molecular Tomography are already used to run quantitative whole body biodistribution and become more useful with the numerous ATMP developments like stem cells, recombinant proteins. Other applications could be envisaged: the imagery technologies could be useful to validate the right injection site and the injected amount when the administration has to be done in the targeted organs (liver, lung by inhalation). As bioluminescence is dependent on the oxygen and ATP amounts in the tissues, it could be used to detect hypoxia and oxidative stress and evaluate changes in metabolic and neuronal activities. The major limit is that these technologies are adapted to small species rodents, because of the light signal absorption by the tissues, whereas they could have an interest in large animals. For instance, the assessment of gastrointestinal function in dogs or monkeys is not easy, and fluorescent or bioluminescent substances used as gastrointestinal function markers could be interesting to run these kinds of investigations. In conclusion, bioluminescence and fluorescence are tools with several potential applications in rodents' preclinical safety studies. doi:10.1016/j.vascn.2012.08.074
How to address metabolite toxicity and species or organ specific toxicity or in vitro? Annie E. Otto-Bruc, David Piwnica, Chantal Rein, Fabienne Sabatier CEREP, Celle L'Evescault, France A xenobiotic that is relatively non-toxic may be metabolized by the liver into toxic metabolites (metabolic activation), which can travel to distant organs (e.g. peripheral neurons, brain), where they cause toxic effects. The opposite is also true: a metabolite can be less toxic than the parent compound. Aflatoxin B1 and Terfenadine are respectively examples of both scenarios. Using IdMOC plates*, we cultured primary cells representing different tissues (kidney, endothelium, immune, lung) in the absence or presence of human hepatocytes to assess respectively drug and related metabolites cytotoxicity. We used Aflatoxin B1 and Terfenadine to validate the technology. Cytotoxicity was measured for each primary cell by the ATP production as an indicator of cell metabolism. The experiments with cocultures of human hepatocytes (metabolically-competent cells)
and CHO cells (metabolically-non-competent cells) confirmed that Aflatoxin B1 requires metabolic activation to be cytotoxic and Terfenadine is metabolically inactivated. Cerep has validated the IdMOC technology to assess metabolismmediated cytotoxicity on primary cells that represent multiple organs. Compound toxicity is also not equivalent in all tissue or all species; we have reproduced the model presented by the NTP program** and applied it to a few drugs. *Integrated discrete Multiple Organ Culture (IdMOC), developed by Dr. Albert Li, Ph.D. of Advanced Pharmaceutical. Sciences, Inc., is a novel technology allowing the culturing of cells from multiple organs in the same culture dish. **Compound Cytotoxicity Profiling Using Quantitative HighThroughput Screening. Menghang Xia et al. Environ Health Perspect 116:284–291 (2008). doi:10.1016/j.vascn.2012.08.075
Direct effects of arsenic trioxide on action potentials in isolated cardiac tissues: Importance of the choice of species, type of cardiac tissues and perfusion time Hua Rong Lu, Eddy Vlaminckx, Jutta Rohrbacher, Frank Cools, Gallacher J. Gallacher Janssen, Pharmaceutical Companies of Johnson and Johnson, Beerse, Belgium Introduction: The aim of the present study was to evaluate direct effects of arsenic trioxide-induced changes in action potentials (AP) in isolated cardiac tissues and investigate if species- and tissuespecific difference and perfusion-time could play an important role in arsenic-induced acute long APD/QT. Methods and results: Acute electrophysiological effects of arsenic were measured in isolated cardiac tissues from four different species using a micro-electrode technique and on the hERG current from HEK293 cells. Arsenic (perfusion at 10 μM after 30 to 95 min) significantly prolonged duration of APD90, increased triangulation of AP and elicited early afterdepolarizations (EADs) in only isolated guineapig and dog Purkinje fibers but not from the rabbit or pig Purkinje. Arsenic-induced long APD90, increases in triangulation and EADs were not observed in papillary muscles of guinea-pigs and rabbits. Arsenic from 0.1 μM to 10 μM with the standard perfusion-time of 15 min interval (per dose) did not induce acute effects on APD90, triangulation and EADs in isolated guinea-pig Purkinje fibers. Furthermore, arsenic up to 10 μM did not directly block hERG currents. Conclusion: The present study demonstrates that species, cardiac tissue-type and perfusion time all have an important role in acute electrophysiological effects induced by arsenic on APD90 and induction of EADs in vitro. Key words: APD (Action potential duration), Triangulation, Long QT syndrome (LQT), Species, Tissue-specific difference, Perfusion time. doi:10.1016/j.vascn.2012.08.076
Does terfenadine-induced VT/VF directly relate to its QT prolongation and Torsades de Pointes? An N. Hermans, Hua Rong Lu, David J. Gallacher Janssen Research & Development, Beerse, Belgium Background: We investigated the proarrhythmic effects of terfenadine in isolated rabbit hearts, arterially-perfused left ventricular
Two novel approaches: Shifting the drug development paradigm Matthew S. Coffee, Javelyn R. Evans, David T. Merriman, Brian M. Roche
179
A retrospective analysis of gastrointestinal adverse events impacting on drug development Jean-Pierre Valentin, A. Al-Saffar, L. Ewart, J. Glab, L. Marks, W.S. Redfern, S. Roberts, T.G. Hammond
Battelle, Columbus, OH, United States AstraZeneca, Safety Assessment, Macclesfield, United Kingdom Over the last 10 years, there has been a significant decline in the number of new drugs being submitted to regulators, while over the same period, the number of regulatory approvals has also declined. This is seen as a result of the ever-increasing cost of drug development. A new drug-development model needs to emerge in order for drug-discovery programs to remain viable into the future. This new model's priorities must include better nonclinical testing, more collaborations or partnerships, and earlier involvement from the research and development provider during the drug discovery phase. This will allow the pharmaceutical industry to make more informed decisions regarding safety and efficacy sooner in the drugdevelopment process. Two approaches to meet this need are being evaluated to develop biobetters and to repurpose failed biologic drugs. Both approaches will ultimately increase the number of medicines available to consumers at a lower cost, thereby shifting the drug-development paradigm in an attempt to revitalize the drug industry. doi:10.1016/j.vascn.2012.08.070
The development of an in vitro safety assay utilising human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) with Multi/micro-electrode array (MEA) technology Amy M. Nicks, Yi Cui, Nick McMahon, Arun Sridhar
We conducted a literature and AZ portfolio review to assess the incidence, prevalence and impact of gastrointestinal (GI) adverse events (AEs) on drug development. In the last few decades, only a small proportion of drugs have been discontinued from development or withdrawn from the market due to GI-AEs (2–6%). However, data from the 2010 ‘DIA Daily Alert’ showed that GI-AEs accounted for 11% of delayed or halted clinical trials; an increase from previous years and a level identical to hepatotoxicity. Furthermore, GI-AEs featured significantly in serious Adverse Drug Reactions (ADRs; 14–23%) and label ADRs (67%). A PharmaPendiumTM search showed that 13% of clinical observations reported were GI disorders, the highest ranking for all organ systems. AEs, reported with 104 marketed drugs in Japan, showed a high incidence of GI-AEs (nausea and vomiting N diarrhoea N anorexia N stomach ache N constipation). Some plausible associations with preclinical GI findings were reported (e.g. intestinal transport, gastric emptying time, orgastric juice secretion vs. anorexia). An AstraZeneca portfolio review of the Phase I single ascending dose clinical trial AE profiles showed a high incidence of high stringency GI-AEs (38% of 95 drugs); an evaluation of the predictive value of nonclinical models is on-going. Ultimately, GI-AEs can be dose limiting preclinically and/or clinically. This affects patient compliance, and can alter the PK profile of drugs. This review suggests that the evaluation of GI liability should be carefully considered prior to human administration.
doi:10.1016/j.vascn.2012.08.072 GlaxoSmithKline, Ware, United Kingdom Adverse cardiac electrophysiological effects are common causes of preclinical drug failure. Multi-micro electrode array (MEA) technology with human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) has the potential to offer a high throughput, more humanlike predictive cardiac electrophysiological model. These experiments identify some of the optimal conditions for this novel assay. The hiPSCCMs were plated at 8000 cells/μl in 10 μl onto MEAs and the extracellular field potential of the electrically connected monolayer that beats spontaneously was recorded. The field potential duration (FPD) and beating rate of hiPSC-CMs were measured from an environmental chamber and compared to a dry incubator. These parameters were also measured in the presence of 0.1% DMSO or E4031 in the dry incubator. The rate of hiPSC-CMs was higher and more variable in the environmental chamber than in the dry incubator. 0.1% DMSO (n = 4) decreased rate by 8%, increased FPD by 6% and FPDc by 8% from baseline values of 0.6 Hz, 761 ± 31 ms and 895 ± 37 ms, respectively, these were not statistically significant (p N 0.05). Preliminary results showed that E-4031 prolonged the FPD at 30 nM and 100 nM by 23% and 34% respectively. To achieve a stable beating rate over time a dry incubator was used; this highlighted the sensitivity of hiPSC-CMs to temperature and CO2. Time dependent changes were seen in 0.1% DMSO; however, they were not significant. E-4031 showed concentration dependent changes confirming its effect of QT prolongation in the clinic (Okada Y et al, 1996). MEA technology and hiPSC-CMs demand further pharmacological validation to define their suitability for predicting clinical effects. doi:10.1016/j.vascn.2012.08.071
Assessment of the concordance between nonclinical repolarization assays and clinical measure of QT interval prolongation John Koernera, B. Darpob, M. Skinnerc, H.M. Vargasd, J. Willarda, S. Pettite, J.P. Valentinc a
U.S. FDA, Silver Spring, MD, United States Pharmaceutical Consultant, Sweden c AstraZeneca, Macclesfield, United Kingdom d Amgen, Inc, Thousand Oaks, CA, United States e HESI on behalf of the Pro-Arrhythmia Model Committee, Washington, DC, United States b
Drug-induced QT interval prolongation and Torsades de Pointes (TdP) remain serious public health issues in bringing safe new pharmaceuticals to the market place. Under the auspices of ILSI Health and Environmental Sciences Institute (HESI), a consortium including representatives from pharmaceutical companies, regulatory agencies and opinion leaders from the scientific and medical research communities has been initiated. The objectives are: to assess the concordance between nonclinical repolarization assays and clinical measures of QT interval prolongation; establish the sensitivity, specificity and predictivity of nonclinical assays towards the clinical outcome; and ultimately to identify compounds that do not show evidence of concordance. Four approaches are pursued and will be summarised: 1. Analysis of the concordance of data extracted from thorough clinical QT studies and nonclinical studies (i.e., in vitro hERG assay, action potential duration in isolated cardiac tissue, and in vivo QTc studies) submitted to the U.S. Food and Drug Administration to support marketing approval of new pharmaceuticals (N 70 drugs in the database). 2. Attempts to include Phase I clinical QTc data to the above analysis.
180
Abstracts
3. Review of the literature using bioinformatics and text mining tools (N120 drugs considered). 4. Retrospective quantitative analysis of the 2001 ILSI-HESI sponsored programme of work (Hanson et al., 2006) involving 12 drugs with clear or no associations with QTc prolongation and TdP. doi:10.1016/j.vascn.2012.08.073
Which possible applications for bioluminescence and fluorescence in preclinical safety studies? Anne Maurin, Eric Martel, Emmanuel Bracq, Laura Brulle, Pascal Champeroux, Serge Richard CERB, Baugy, France Bioluminescence and fluorescence are more and more used to assess the pharmacological activities of NCE in vivo. Their main advantages are they are not invasive, harmlessness and allow establishing kinetics on the same animal as illustrated with our results of orthotopic, colorectal, pancreatic and bones cancers in mice. In preclinical development, Fluorescence and Bioluminescence Molecular Tomography are already used to run quantitative whole body biodistribution and become more useful with the numerous ATMP developments like stem cells, recombinant proteins. Other applications could be envisaged: the imagery technologies could be useful to validate the right injection site and the injected amount when the administration has to be done in the targeted organs (liver, lung by inhalation). As bioluminescence is dependent on the oxygen and ATP amounts in the tissues, it could be used to detect hypoxia and oxidative stress and evaluate changes in metabolic and neuronal activities. The major limit is that these technologies are adapted to small species rodents, because of the light signal absorption by the tissues, whereas they could have an interest in large animals. For instance, the assessment of gastrointestinal function in dogs or monkeys is not easy, and fluorescent or bioluminescent substances used as gastrointestinal function markers could be interesting to run these kinds of investigations. In conclusion, bioluminescence and fluorescence are tools with several potential applications in rodents' preclinical safety studies. doi:10.1016/j.vascn.2012.08.074
How to address metabolite toxicity and species or organ specific toxicity or in vitro? Annie E. Otto-Bruc, David Piwnica, Chantal Rein, Fabienne Sabatier CEREP, Celle L'Evescault, France A xenobiotic that is relatively non-toxic may be metabolized by the liver into toxic metabolites (metabolic activation), which can travel to distant organs (e.g. peripheral neurons, brain), where they cause toxic effects. The opposite is also true: a metabolite can be less toxic than the parent compound. Aflatoxin B1 and Terfenadine are respectively examples of both scenarios. Using IdMOC plates*, we cultured primary cells representing different tissues (kidney, endothelium, immune, lung) in the absence or presence of human hepatocytes to assess respectively drug and related metabolites cytotoxicity. We used Aflatoxin B1 and Terfenadine to validate the technology. Cytotoxicity was measured for each primary cell by the ATP production as an indicator of cell metabolism. The experiments with cocultures of human hepatocytes (metabolically-competent cells)
and CHO cells (metabolically-non-competent cells) confirmed that Aflatoxin B1 requires metabolic activation to be cytotoxic and Terfenadine is metabolically inactivated. Cerep has validated the IdMOC technology to assess metabolismmediated cytotoxicity on primary cells that represent multiple organs. Compound toxicity is also not equivalent in all tissue or all species; we have reproduced the model presented by the NTP program** and applied it to a few drugs. *Integrated discrete Multiple Organ Culture (IdMOC), developed by Dr. Albert Li, Ph.D. of Advanced Pharmaceutical. Sciences, Inc., is a novel technology allowing the culturing of cells from multiple organs in the same culture dish. **Compound Cytotoxicity Profiling Using Quantitative HighThroughput Screening. Menghang Xia et al. Environ Health Perspect 116:284–291 (2008). doi:10.1016/j.vascn.2012.08.075
Direct effects of arsenic trioxide on action potentials in isolated cardiac tissues: Importance of the choice of species, type of cardiac tissues and perfusion time Hua Rong Lu, Eddy Vlaminckx, Jutta Rohrbacher, Frank Cools, Gallacher J. Gallacher Janssen, Pharmaceutical Companies of Johnson and Johnson, Beerse, Belgium Introduction: The aim of the present study was to evaluate direct effects of arsenic trioxide-induced changes in action potentials (AP) in isolated cardiac tissues and investigate if species- and tissuespecific difference and perfusion-time could play an important role in arsenic-induced acute long APD/QT. Methods and results: Acute electrophysiological effects of arsenic were measured in isolated cardiac tissues from four different species using a micro-electrode technique and on the hERG current from HEK293 cells. Arsenic (perfusion at 10 μM after 30 to 95 min) significantly prolonged duration of APD90, increased triangulation of AP and elicited early afterdepolarizations (EADs) in only isolated guineapig and dog Purkinje fibers but not from the rabbit or pig Purkinje. Arsenic-induced long APD90, increases in triangulation and EADs were not observed in papillary muscles of guinea-pigs and rabbits. Arsenic from 0.1 μM to 10 μM with the standard perfusion-time of 15 min interval (per dose) did not induce acute effects on APD90, triangulation and EADs in isolated guinea-pig Purkinje fibers. Furthermore, arsenic up to 10 μM did not directly block hERG currents. Conclusion: The present study demonstrates that species, cardiac tissue-type and perfusion time all have an important role in acute electrophysiological effects induced by arsenic on APD90 and induction of EADs in vitro. Key words: APD (Action potential duration), Triangulation, Long QT syndrome (LQT), Species, Tissue-specific difference, Perfusion time. doi:10.1016/j.vascn.2012.08.076
Does terfenadine-induced VT/VF directly relate to its QT prolongation and Torsades de Pointes? An N. Hermans, Hua Rong Lu, David J. Gallacher Janssen Research & Development, Beerse, Belgium Background: We investigated the proarrhythmic effects of terfenadine in isolated rabbit hearts, arterially-perfused left ventricular