Journal of pharmacological and toxicological methods最新文献

{"title":"Evaluation of temperature and trafficking modulators on hERG current via high-throughput automated patch clamp","authors":"Maria Giustina Rotordam,&nbsp;Wyatt Hall,&nbsp;Nadine Becker,&nbsp;Soren Friis,&nbsp;Tim Strassmaier,&nbsp;Alison Obergrussberger,&nbsp;Michael George,&nbsp;Niels Fertig","doi":"10.1016/j.vascn.2026.108510","DOIUrl":"10.1016/j.vascn.2026.108510","url":null,"abstract":"<div><div>The human Ether-à-go-go-Related Gene (hERG) channel is a voltage-gated potassium channel crucial for cardiac repolarization. Mutations in the hERG channel can lead to long QT syndrome, a condition associated with potentially fatal arrhythmias. Additionally, a wide variety of drugs can block the hERG channel, which may also trigger life-threatening arrhythmias. Consequently, all new pharmaceuticals must be evaluated for their effects on hERG currents using the patch clamp technique.</div><div>Automated patch clamp (APC) systems are now widely adopted—if not standard—in laboratories conducting cardiac safety assessments on hERG and other cardiac ion channels. To ensure reliable APC data, cell culture conditions for cells overexpressing the hERG channel must be carefully optimized. In our study, we explored various culture conditions to improve experimental outcomes.</div><div>We systematically assessed different variables, including the use of trafficking and hERG channel blockers in the culture medium, and employed an automated cell harvesting protocol to compare how these conditions influenced hERG current measured by APC (SyncroPatch 384). Notably, culturing cells at 32 °C for 24 h before the experiment significantly increased current amplitude by around 200 pA (442 pA, <em>n</em> = 63 at 37 °C vs. 654 pA, <em>n</em> = 150 at 32 °C).</div><div>We also examined the effects of culturing cells at 37 °C in the presence of the trafficking inhibitor nocodazole and the hERG blocker bepridil. Cells were grown in 384-well plates under these different conditions and then harvested using the patch clamp robot for analysis. Our results showed that a 24-h incubation with nocodazole doubled the hERG tail current compared to vehicle control. In contrast, bepridil treatment caused a substantial reduction in hERG tail current. Moreover, pre-incubation with bepridil shifted its IC₅₀ in follow-up assays by nearly 20-fold compared to vehicle, while nocodazole had no impact on bepridil's IC₅₀.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108510"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effectively de-risking adverse central nervous system effects of novel therapeutics: A case study discussing the confidence provided by a combination of in vitro and in vivo screening assays with a novel antitumor agent","authors":"Kate M. Sanders,&nbsp;Julia Luginbuhl,&nbsp;Sowjanya Prayaga,&nbsp;Alex Nguyen,&nbsp;Dave Adams,&nbsp;Shelby Steele,&nbsp;Michael Flagella,&nbsp;Zhengping Wang","doi":"10.1016/j.vascn.2026.108439","DOIUrl":"10.1016/j.vascn.2026.108439","url":null,"abstract":"<div><div>Safety scientists are commonly tasked with developing screening cascades to derisk CNS safety liabilities of novel therapeutics early in the drug discovery process. As new approach methodologies and updates to the ICH S7A guidance gain traction, safety pharmacologists may consider further evaluating the advantages and limitations of screening assays. Here, we discuss one in vivo and three in vitro assays used to evaluate CNS effects of a RAS(ON) inhibitor as part of a case study examining the level of confidence their results provide for CNS safety. Functional observational battery (FOB) assessments in mice showed no CNS effects at brain C_max concentrations approximately 19-fold higher than that associated with antitumor activity in an intracranial (IC) CDX mouse model. Additionally, there were no effects on iPSC neurons uncovered during microelectrode array assessment, nor any significant EC/IC₅₀ activity with relevant CNS receptors during two <em>in vitro</em> binding assays relative to efficacious tumor C_max. The lack of activity in the <em>in vitro</em> and in vivo studies, in addition to what is already known about RAS(ON) inhibitors in the clinic, suggest that this new compound is unlikely to elicit acute CNS side-effects. Additional studies to further derisk a CNS-penetrative compound could include testing the compound in a nonrodent species and evaluating extended duration of exposure to the compound during in vitro and in vivo assays. These additional assessments could further derisk and increase confidence for CNS safety regardless of the intended target or indication.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108439"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730172","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chamber-specific hiPSC-CMs to investigate drug-induced atrial cardiotoxicity","authors":"Cristina Altrocchi,&nbsp;Anneleen Dierckx,&nbsp;Mohamed Kreir,&nbsp;David G. Gallacher","doi":"10.1016/j.vascn.2026.108500","DOIUrl":"10.1016/j.vascn.2026.108500","url":null,"abstract":"<div><div>Cardiotoxicity remains one of the primary causes of drug withdrawal from the market and poses significant challenges during preclinical drug discovery and development. Focusing on supraventricular rhythm changes, drug-induced atrial fibrillation (AF) and alterations in heart rate (HR) are critical concerns, especially in the oncological space, that warrant closer investigation during safety pharmacology/toxicology studies. AF itself is a complex condition marked by diverse structural and electrical remodelling within the atrial tissue, linked to several risk factors (age, hypertension, etc.) making it challenging to study and model, especially <em>in vitro</em>.</div><div>In this study, we explored the utility of human-induced pluripotent stem cell-derived atrial cardiomyocytes (hiPSC-CMs) as an <em>in vitro</em> platform to evaluate the potential cardiotoxic effects of atrial-specific compounds, particularly focusing on drug-induced heart rate (HR) changes and AF. To validate this approach, we tested multiple sources of atrial<em>-like</em> hiPSC-CMs and compared their baseline electrophysiological profiles, including calcium transient duration, beating rate and amplitude, against those of primarily ventricular<em>-like</em> hiPSC-CMs. We also compared the response of these cells to atrial-specific compounds involved in HR variations as well as AF in the clinic, via functional assays (calcium transients &amp; MultiElectrode Array). Finally, transcriptional (high-throughput RNA sequencing) and proteomics profiles were also compared.</div><div>Our findings highlighted differences in drug-response between atrial and ventricular hiPSC-CMs, such as their respective response to β1-adrenergic receptor stimulation by dobutamine, and the pro-arrhythmic effects of the hERG blocker dofetilide, reaffirming the relevance of chamber-specific models for studying atrial cardiotoxicity. This <em>in vitro</em> approach demonstrates promise as a tool to better predict atrial-specific cardiotoxic risks. Ultimately, leveraging <em>in vitro</em> models can improve preclinical screening pipelines and help mitigate safety concerns during the drug discovery process, supporting the reduction of <em>in vivo</em> animal studies.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108500"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730260","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preclinical evaluation of the efficacy and safety of temozolomide in a murine glioblastoma model","authors":"Sonia Goineau,&nbsp;Elise Esneault,&nbsp;Laurie Genest,&nbsp;Sabrina Djemmali,&nbsp;Philippe Guillaume","doi":"10.1016/j.vascn.2026.108450","DOIUrl":"10.1016/j.vascn.2026.108450","url":null,"abstract":"","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108450"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A CellDrum™ based human aortic smooth muscle - Endothelial cell co-culture for quantifying the efficacy of tensile cell force-inducing and relaxing agents","authors":"Karya Uysal,&nbsp;Ipek S. Firat,&nbsp;Till P. Creutz,&nbsp;Gerhard M. Artmann,&nbsp;Aysegül Temiz Artmann","doi":"10.1016/j.vascn.2026.108486","DOIUrl":"10.1016/j.vascn.2026.108486","url":null,"abstract":"<div><div>For drug development it is essential to test cellular tensile forces acting in cell layers and tissues. In the past, these tests were often carried out on animal vascular ring explants. We have developed a quantitative <em>in vitro</em> co-culture model for primary human aortic smooth muscle cells (hAOSMC) and endothelial cells (hAOEC), based on the CellDrum™ technology. This allows for quantifying the efficacy of substances inducing or relaxing cell tensile forces. Cell cultures on flexible CellDrum™ membranes together with the Cell Force Analyzer (CFA_ref) were used for quantifying tensile forces induced by Norepinephrine (NA) and the calcium ionophor Ionomycin. Primary hAOSMC were seeded on the CellDrumTM membranes and allowed to differentiate for eight days before primary hAOEC (ATCC, VA, USA) were added. Co-cultures were left to mature for an additional two days. The layered, confluent structure of the co-culture was verified via confocal imaging. These revealed the uniformity of the co-culture. Images were processed in ImageJ by overlaying the channels onto the antibody-stained images. NA and IO effects were tested for force induction or relaxation in both SMC monolayers and SMC-EC co-cultures. Norepinephrine induced a significant hAOSMC monolayer tensile tension increment by 16.9% from control (EC50: 100.70 nM). Co-cultures of hAOSMC and hAOECs in contrary led to a tensile tension relaxation of 12.6% compared to control (EC50: not applicable). The calcium ionophor Ionomycin, in comparison, showed a 10.9% tensile tension increment (EC50: 11.44 nM) with hAOSMC monolayers alone. With co-cultures, however, its contractile effect was up to 60% higher than control (EC50: &lt;1 nM). Both substances have the expected effect on hAOSMC monolayers. The interaction of the cells in the co-culture instead reveals an opposite effect in both cases. The ionomycin effect of the co-culture is considerably enhanced. Norepinephrine (NA), instead, leads to relaxation. The results are irrefutable: the effects of drugs vary significantly between cell co-cultures and monolayers. We attribute the relaxing effect of NA to the stimulation of α₂-adrenoceptors of the endothelial cells, which leads to the release of nitric oxide (NO), a vasodilator.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108486"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730634","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"New approach methodologies (NAMs) in safety pharmacology: Ready for prime time?","authors":"Will S. Redfern,&nbsp;Lorna S. Ewart,&nbsp;Chris E. Pollard,&nbsp;Kevin Snyder","doi":"10.1016/j.vascn.2026.108487","DOIUrl":"10.1016/j.vascn.2026.108487","url":null,"abstract":"<div><div>The FDA's ‘Roadmap to reducing animal testing in preclinical studies’ (Anon., 2025) sets out an ambitious and clear objective “to make animal studies the exception rather than the norm for pre-clinical safety/toxicity testing” within 3–5 years from now, starting with monoclonal antibodies. Animal studies will progressively be replaced by a combination of in vitro and in silico New Alternative Methods (NAMs). Until now, NAMs have largely been deployed for hazard detection prior to in vivo testing. To assess their current state of readiness for promotion to a role in pre-FTIH safety assessment, we undertook a preliminary, qualitative gap analysis of available NAMs across the five organ systems of interest to safety pharmacology. Each organ system was rated ‘High’, ‘Medium’ or ‘Low’ (relative terms) according to maturity of the technology, pharmacological validation, reproducibility/standardization initiatives, GLP/.</div><div>‘GLP-analogous’ status and current regulatory acceptance. The organ system with the highest rating was ‘CVS: QT’, scoring ‘High’ across all five measures. All the other organ systems (CVS-other, central nervous system, respiratory, gastrointestinal and renal) were rated ‘Low’ across ‘reproducibility/standardization initiatives’, ‘GLP status’ and ‘regulatory compliance’, with mixed ratings against ‘maturity of the technology’ and ‘pharmacological validation’. In considering deploying NAMs in place of in vivo studies, the following became apparent. 1. The distinction between safety pharmacology and toxicology assays becomes blurred, with some assays reporting both functional and pathological endpoints. 2. The spectrum of measurements will not completely overlap with current in vivo studies, leaving some in vivo measurements unaddressed and performing some assessments that are not currently done in vivo. 3. To maintain adequate safety assessment for FTIH trials, NAMs should demonstrate improved predictive value for human clinical outcomes rather than merely benchmarking to current <em>in vivo</em> models. In conclusion, although in vitro and in silico NAMs exist across the spectrum of in vivo measurements currently undertaken in safety pharmacology, their current context of use is not as pre-FTIH pivotal assays. The safety pharmacology community should work closely together to agree on how to address the FDA's roadmap.<ul><li><span>1.</span><span><div>Anon. (2025) Roadmap to reducing animal testing in preclinical safety studies. <span><span>https://www.fda.gov/media/186092/download</span><svg><path></path></svg></span>.</div></span></li></ul></div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108487"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730647","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The application of Bayesian detection of risk using inference on blinded safety (BDRIBS) to the Phase 3 GLISTEN study","authors":"Sadia Halim,&nbsp;Luca Grassano,&nbsp;Houssem Khanfir,&nbsp;Julie Rouette,&nbsp;Ivana Lazic,&nbsp;Sugato Das,&nbsp;Naomi Givens","doi":"10.1016/j.vascn.2026.108484","DOIUrl":"10.1016/j.vascn.2026.108484","url":null,"abstract":"<div><div>Early identification of adverse events (AEs) associated with investigational products is of paramount importance. The BDRIBS method can help detect potential safety signals in ongoing blinded trials, as recommended by the Food and Drug Administration, and support targeted AEs for unblinded review. Using background rates of AEs, the BDRIBS method evaluates accumulated evidence (blinded) to infer the relative risk (RR) of AEs in the investigational treatment arm versus control. Prior information is combined with collected data to form a posterior distribution from which inference is made; thus the choice of prior information is critical. Here we describe a blinded evaluation of the RR of AEs between linerixibat, an ileal bile acid transporter inhibitor (IBATi), and placebo in the Phase 3 GLISTEN study. The analysis included GLISTEN participants who received ≥1 dose of treatment from baseline to Week 24. Targeted AEs were gallstones and cholecystitis (considered a theoretical risk with IBATi based on published evidence). As the incidence of these AEs was rare in GLISTEN (gallstones: <em>n</em> = 0, cholecystitis: <em>n</em> = 2), this abstract focuses on cholecystitis only. Background rates of cholecystitis AEs were identified in patients with primary biliary cholangitis using US claims data. The posterior probability (P) of RR &gt;1 for an AE for linerixibat versus placebo was assessed. This threshold was selected as any elevation in AE events would trigger unblinded safety review. To account for uncertainty in background event rate, sensitivity analysis evaluated P(RR &gt; 1) under alternative event rates (5th and 95th percentiles) versus GLISTEN data. Background rates of cholecystitis were 13.73/1000 person-years (PY). The P(RR &gt; 1) of cholecystitis in the linerixibat group versus placebo was 50.28%, suggesting no increased risk. In the sensitivity analysis, assuming a conservative background event rate of 10.51/1000 PY, with two observed events in GLISTEN, the P(RR &gt; 1) was 60.70%. For P(RR &gt; 1) to be ≥80% with the same background event rate, ≥4 cases would have to have been observed in GLISTEN versus the two reported.</div><div>The BDRIBS analysis of GLISTEN suggests no observed increase risk of cholecystitis with linerixibat versus placebo. Sensitivity analyses with more conservative background rates supported these findings.</div><div>Funding: GSK (212620).</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108484"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730669","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modified neurobehavioral test with telemetered hemodynamic and left ventricular pressure assessment to detect CNS biomarker after oral dose of chlorpromazine in rats","authors":"Jennifer Shaffer,&nbsp;Nick Adkins,&nbsp;Brittany Balser,&nbsp;Kevin Norton","doi":"10.1016/j.vascn.2026.108518","DOIUrl":"10.1016/j.vascn.2026.108518","url":null,"abstract":"<div><div>The ICH S7A guidelines outline nonclinical safety pharmacology assessment, including the central nervous system (CNS). Drugs targeting the CNS present potential investment risk as only 8.2% of these drugs progress to market in comparison to 15% of treatments targeting other indications. Currently, the Irwin assessment is a standard Tier I test determining acute effects of a compound on rodent CNS activity. While useful in predicting overt effects, it is subjective and proves difficult in determining changes in cognitive function. The addition of effective biomarkers to the Irwin may illuminate latent neurotoxicities and indicate a need for further supplemental testing.</div><div>The effects of chlorpromazine (a drug with known sedative effects targeting both CNS and cardiovascular systems) were evaluated by the Irwin assessment (5 timepoints) in 18 Sprague Dawley rats. To assess correlation between CNS liabilities and effects on the circulatory system, blood pressure and left ventricular pressure parameters, heart rate, body temperature, and activity were collected continuously for 22 h following oral dosing.</div><div>Administration of chlorpromazine resulted in motor activity changes, sedation and increased pain response in conjunction with hemodynamic changes (increases in heart rate, systolic, diastolic and mean arterial blood pressure), increased body temperature and activity, left ventricular pressure changes (increases in dP/dt[min] and decreases in dP/dt[max]).</div><div>The additional telemetered data were able to identify the drug onset and duration while the Irwin assessment was able to illustrate the behavioral changes associated with the drug pharmacodynamics. Information gathered from the additional endpoints allows more in depth insight to the behavioral changes in the Tier I CNS assessment as well as the potential benefit of supplemental testing.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108518"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730718","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of cardiovascular monitoring methodology on sensitivity and reliability in non-rodent toxicology studies","authors":"Jean Pierre Valentin,&nbsp;Todd Bourcier,&nbsp;Xuan Chi,&nbsp;Annie Delaunois,&nbsp;C. Michael Foley,&nbsp;Kim A. Henderson,&nbsp;Pierre Lainee,&nbsp;Derek J. Leishman,&nbsp;Dingzhou Li,&nbsp;Emma Pawluk,&nbsp;Michael K. Pugsley,&nbsp;Sridharan Rajamani,&nbsp;Christopher P. Regan,&nbsp;Michael G. Rolf,&nbsp;Rebecca Ross,&nbsp;Eric I. Rossman,&nbsp;Stephen D. Tichenor,&nbsp;Inmaculada C. Villar,&nbsp;Todd A. Wisialowski,&nbsp;Jean Wu,&nbsp;Hugo M. Vargas","doi":"10.1016/j.vascn.2026.108476","DOIUrl":"10.1016/j.vascn.2026.108476","url":null,"abstract":"<div><div>Cardiovascular (CV) parameters such as blood pressure (BP), electrocardiogram (ECG), and heart rate (HR) are routinely assessed in non-rodent toxicology studies, yet measurement quality varies by methodology—namely telemetry (implanted or jacketed) versus restraint-based techniques. This analysis examined how measurement methods affect baseline CV values and sensitivity to drug-induced effects. Data from three sources were analyzed: 495 toxicology studies from seven pharmaceutical companies (2015–2023), FDA recently approved drugs (47 NCEs, 26 NBEs in 2022–2023), and two major CROs (2020−2023). Studies involved dogs, minipigs, or non-human primates with dosing durations up to 52 weeks. Additional literature, proprietary data, and questionnaire informed baseline comparisons and statistical and pharmacological sensitivity. In our toxicology datasets, ECG was commonly collected, whereas BP was not. The selection of ECG and BP recording methods was mostly sponsor-dependent. Telemetry was predominantly used in short-duration studies; restraint-based snapshot methods were common in longer studies. Restraint consistently increased baseline BP and HR values and variability. QTc correction methods under restraint were inconsistent across sponsors/CROs. Statistical analysis was common with telemetry but infrequent under restraint. Case examples show that telemetry detected clinically-relevant BP or QTc changes; however, restraint methods usually did not. These findings demonstrate that restraint methods increase baseline values and variability, reducing pharmacological and statistical sensitivity to detect CV effects. Current practices appear driven by sponsor convention as well as scientific rationale. As ICH S7A/S7B evolve, re-evaluation of CV monitoring methods is warranted to enhance scientific rigor, regulatory alignment, and 3Rs compliance in nonclinical safety assessment.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108476"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148731188","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Examining moxifloxacin QTc sensitivity in the canine to support ICH E14/S7B Q&A compliant studies using a parallel ascending and an ascending dosing scheme","authors":"Ioannis Christodoulou,&nbsp;Ninette Andersen,&nbsp;Nacera Mellal,&nbsp;Jared Slain,&nbsp;Anson Philips,&nbsp;Steve Denham,&nbsp;Matthew St. Peter,&nbsp;Mohamed Farrag,&nbsp;Holly Madison,&nbsp;Tomas Mow,&nbsp;Louise Bang-Lauritsen,&nbsp;Sophie Strindberg,&nbsp;Anette Grossi,&nbsp;Michael Scott","doi":"10.1016/j.vascn.2026.108465","DOIUrl":"10.1016/j.vascn.2026.108465","url":null,"abstract":"<div><div>Following adoption of the ICH E14/S7B Q&amp;A best practice guidance, industry position papers and reports have predominantly described the sensitivity and power of the Latin square model to detect QT changes. Alternative dose designs may be needed for test items with prolonged half lives, where a cross-over design is not feasible. The current study objective was to determine QTc sensitivity with moxifloxacin in telemetry implanted canines using a parallel ascending or an ascending dose design. All animal data was collected on IACUC approved protocols; 6 control animals received vehicle on Days 1, 3, 10, and 17, while 5 or 6 animals got vehicle on Day 1 and moxifloxacin at 10, 30, and 90 mg/kg by oral gavage on Days 3, 10, and 17, respectively. Parallel ascending design analysis compared both groups (<em>n</em> = 5/6) on Days 3, 10 and 17. Ascending design analysis compared treatment group data on Days 3, 10, and 17 with control data on Day 1 in the same animals (n = 5). A full pharmacokinetic assessment was conducted for plasma concentration-QTc modeling, based on the ascending design. With both dose designs, QTc changes were observed with statistically significant maximal QTc increases of 10, 24 and 50 ms (parallel ascending design) or 10 (trend only), 22 and 46 ms (ascending design) at 10, 30 and 90 mg/kg, respectively. Thus, both study designs detected moxifloxacin-related QTc prolongation in a dose-dependent manner, with sensitivity (LSD) values of 9.7 ms (RMSE = 5.7) for the parallel ascending, and 12.1 ms (RMSE = 8.8) for the ascending design. Finally, based on the double-delta Linear Mixed Effect model using the ascending design, the plasma concentration of moxifloxacin to elicit a 10 ms QTc prolongation was calculated to be 4453 ng/mL (3162 ng/mL unbound). The unbound concentration of 3162 ng/mL was within 2.8-fold of the human critical concentration (1120 ng/mL unbound) known to produce a 10 ms effect in human. In conclusion, our data demonstrate the sensitivity of both the parallel ascending and ascending designs to detect QTc changes, in alignment with the most recent ICH E14/S7B Q&amp;A guidelines.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108465"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148731193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}