Journal of pharmacological and toxicological methods最新文献

{"title":"Ion channel trafficking studies using automated patch clamp technology","authors":"Laura Castelletti,&nbsp;Selena Nola,&nbsp;Manuela Medelin,&nbsp;Daniela Gavello,&nbsp;Valeria Guardiani,&nbsp;Caterina Virginio","doi":"10.1016/j.vascn.2026.108506","DOIUrl":"10.1016/j.vascn.2026.108506","url":null,"abstract":"<div><div>Throughout the drug development process, one of the most common adverse effects leading to drug failure is cardiac arrhythmia. This condition arises from dysfunctions in the activity or expression of various ion channels that shape the cardiac action potential. Moreover, ion channel trafficking is a contributing factor in both inherited and acquired arrhythmias. Unlike the rapid effects of acute ion channel inhibition, disruptions in trafficking typically emerge over longer periods, ranging from several hours to days.</div><div>Several pharmacological agents, such as arsenic trioxide and pentamidine, can induce hERG channel deficiency by interfering with its biogenesis and intracellular trafficking.</div><div>A high-throughput assay for detecting drug-induced inhibition of hERG channel trafficking has been established using the SyncroPatch platform. This assay enables the identification of trafficking defects in recombinant cell lines following overnight drug exposure. The development and validation of the assay involved testing known therapeutic compounds (such as arsenic trioxide, amitriptyline, fluoxetine, pentamidine, amiodarone), and these drugs have been confirmed in this assay to cause hERG channel trafficking defects.</div><div>Briefly, 48 h prior to the experiment, cells resuspended in culture medium were plated in 384-well plates. After 24 h, compounds to be assessed as well as vehicle were added in the 384-well plates. Cells were incubated with compounds for 24 h. The day of the experiment, cells were detached from each well in the incubated assay plate and resuspended in the experimental solution. Evaluation of pre-treatment effects on the hERG current was possible thanks to the SyncroPatch, an automated patch clamp instrument appropriate for studying ion channels. Impairment of hERG channel function due to prolonged exposure of cells to drugs was evaluated measuring the alteration in peak hERG current amplitude following drug treatment compared to vehicle.</div><div>Understanding the mechanisms of drug-induced cardiotoxicity and differences in cardiac safety among drugs is key to developing safer compounds and better preclinical safety tests. The described assay, already adapted in our laboratories to the hNav1.5 sodium channel, highlights its broader applicability, as trafficking defects have been observed in multiple ion channels beyond hERG.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108506"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730745","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":"From surplus to scientific value: Repurposing vascular tissues for organ bath standardization to de-risk vasoactive liabilities","authors":"Laura Coonen,&nbsp;Cristina Altrocchi,&nbsp;Jeroen Tolboom,&nbsp;Hans De Wolf,&nbsp;Mohamed Kreir,&nbsp;David J. Gallacher","doi":"10.1016/j.vascn.2026.108563","DOIUrl":"10.1016/j.vascn.2026.108563","url":null,"abstract":"<div><h3>Introduction</h3><div>The vascular organ bath assay is a well-established classical pharmacology model to characterize vasoactive properties of compounds, offering a platform to elucidate mechanisms of action, optimize dosing, and identify cardiovascular hazards early in the drug selection process and derisking. To address both the regulatory emphasis on detecting drug-induced blood pressure changes and the increasing importance of the 3Rs, repurposed vascular tissue was used to identify key procedural determinants of tissue responsiveness and to support standardization in organ bath studies</div></div><div><h3>Methods</h3><div>Using a linear mixed effects model, we evaluated the impact of procedural variables on quality control test outcomes and established an optimized and standardized protocol prioritizing efficiency, robustness, and maximal tissue use. Comprehensive tissue characterization was achieved by generating dose-response curves for relevant reference compounds including phenylephrine, acetylcholine, serotonin and gallopamil. Finally, the vasoactive properties of pimobendan were investigated and mechanistic experiments were performed to explore its mechanism of action</div></div><div><h3>Results</h3><div>Overall, sedation method was the most limiting factor identified, as isoflurane markedly attenuated acetylcholine induced relaxation after extended storage of the aortic rings. While the set-up did not capture centrally mediated, humoral or chronic blood pressure effects, we were able to effectively reveal direct vasoactive properties of reference compounds with physiological relevance. Successful mechanistic studies were performed, suggesting a potential role for the endothelium and PVAT in pimobendan induced relaxation.</div></div><div><h3>Conclusion</h3><div>These findings validate the use of repurposed tissues for evaluating pressor effects, support assay harmonization, and demonstrate how thoughtful refinement can reconcile regulatory safety needs with the 3Rs principles in mind.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108563"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148458243","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":"Two new approach methodologies (NAMs) predict no clinical QTc prolongation by sisunatovir - Confirmed by TQT study","authors":"Peter Kilfoil,&nbsp;Eric Watt,&nbsp;Todd A. Wisialowski,&nbsp;Ryan Franke","doi":"10.1016/j.vascn.2026.108504","DOIUrl":"10.1016/j.vascn.2026.108504","url":null,"abstract":"<div><div>Sisunatovir, an orally administered inhibitor of the Respiratory Syncytial Virus F-protein, inhibits the hERG channel in vitro (IC50 = 1.8 μM), providing a ~ 15× safety margin over the high clinical exposure (hCmax; 118 nM free). A hERG margin &lt;30× generally indicates increased risk for clinical QT prolongation and the arrhythmia Torsades de Pointes (TdP). Sisunatovir also inhibits the cardiac sodium (Nav1.5) and calcium (Cav1.2) channels at 12.1 μM and 19.8 μM, respectively, which may offset the delayed repolarization imparted by hERG block, limiting QT prolongation.</div><div>We have previously developed and published two NAMs that provide an integrated assessment of multi-ion channel block. Our human induced pluripotent stem cell cardiomyocyte (hIPSC-CM) model, uses rate-corrected action potential duration (cAPD) changes to estimate the concentration associated with clinical 10 ms QTc prolongation. Our in silico model, trained on a library of clinical compounds with known TdP risk, uses ion channel inputs (hERG binding Ki, Nav1.5 and Cav1.2 functional patch IC50's) to simulate voltage and calcium waveforms based on the O'Hara-Rudy ventricular cardiomyocyte model to predict TdP risk. Here we employed these models to assess the repolarization and TdP risks of a novel compound prior to its examination in a Thorough QT (TQT) study.</div><div>The hiPSC-CM model predicted that 30-min sisunatovir exposure would cause a 10 ms increase in clinical QTc at 1.8 μM (&gt;15× hCmax) and that 24-h exposure would cause no clinical QTc prolongation up to 76 μM (&gt;640× hCmax). The in silico model predicted a low risk of TdP at free plasma concentrations ≤1.4 μM (&gt;10× hCmax). These results with preclinical NAMs suggest that sisunatovir is unlikely to result in QTc prolongation or elevated TdP risk at clinically relevant exposures. A TQT study (NCT05878522) confirmed no QTc signal of potential clinical concern. These results demonstrate the utility of NAMs for integrated risk assessment of QTc and TdP.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108504"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730089","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":"Enhancing respiratory safety pharmacology and supporting 3Rs via visualisation of breathing patterns using symmetric projection attractor reconstruction","authors":"Miquel Serna Pascual,&nbsp;Carolyn Lam,&nbsp;Aileen Milne,&nbsp;Mary McElroy,&nbsp;Manasi Nandi","doi":"10.1016/j.vascn.2026.108453","DOIUrl":"10.1016/j.vascn.2026.108453","url":null,"abstract":"<div><div>Recent cross-sector investigation highlighted limited clinical translatability of ICHS7a respiratory core battery. This presents ethical and scientific challenges, as studies may yield limited actionable insight while consuming significant resources—contravening 3Rs principles. Key aspects of lung physiology are conserved between rats and humans, justifying core battery assessment. However, inadequate acclimatisation to plethysmography chambers can introduce stress or exploratory artefacts. Moreover, despite generating high-fidelity time-series data, plethysmography systems utilize just a minute fraction of this e.g. waveform maxima/minima for respiratory rate (RR) and tidal volume calculations. These factors may lead to compromised results, contributing to poor reproducibility and clinical translation. We hypothesised that Symmetric-Projection-Attractor-Reconstruction (SPAR), a waveform-based visualisation technology, could offer a more sensitive assessment of respiratory function. By representing entire high-fidelity breathing waveforms as SPAR-images, it enables objective pattern recognition and assessment of respiratory physiology. We applied this to a rat fibrosis model, with known changes in breathing. Adult male Sprague-Dawley rats (<em>n</em> ≥ 8/group) received bleomycin, bleomycin + nintedanib (standard-of-care), or vehicle. Respiratory data, collected via head-out plethysmography across multiple timepoints, were analyzed using conventional metrics and SPAR. Conventional metrics confirmed known bleomycin effects, including elevated RR and apnoea burden, peaking at days 5–14. In parallel, SPAR revealed distinctive changes (rounding) relating to bleomycin provoked increased breathing-effort, with nintedanib attenuating these alterations. More importantly, SPAR detected high baseline variability within control animals, with some deviating from the characteristic 3-armed images to rounded —suggesting inadequate acclimatisation or subclinical differences. SPAR uses all available respiratory waveform data. It provides a scalable, quantifiable visual representation of lung function that could support better experimental design and cross-study harmonisation. SPAR rounding, with increased breathing-effort observed in this study, is clinically translatable (doi: <span><span>10.1183/23120541.00164-2023</span><svg><path></path></svg></span>). SPAR could support wider feature extraction to discriminate sniffing/grooming, from signals associated with breathing effort for gas exchange. We propose SPAR as a method to justify animal/data inclusion/exclusion, fine-tune input data quality and reduce inter-subject variability, without inadvertent bias introduction. This approach would refine respiratory phenotyping, potentially reducing unnecessary animal use, and strengthen translational relevance—thus aligning with the ethical and scientific goals of ICHS7a and the 3Rs.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108453"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730511","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":"Toxicological profiling of Samrannitra: A traditional Thai Cannabis-based formulation evaluated following OECD guideline 408","authors":"Bhanubong Bongcheewin,&nbsp;Nutputsorn Chatsumpun,&nbsp;Natthinee Anantachoke,&nbsp;Anusak Kijtawornrat,&nbsp;Rungravi Temsiririrkkul,&nbsp;Nakkawee Saengklub","doi":"10.1016/j.vascn.2026.108438","DOIUrl":"10.1016/j.vascn.2026.108438","url":null,"abstract":"<div><div>Samrannitra is a conventional Thai herbal preparation including <em>Cannabis sativa</em> L., traditionally utilized to mitigate insomnia, especially among the elderly. Due to its ability to modulate the central nervous system (CNS), assessing its safety profile is essential for clinical development. This study sought to evaluate the subchronic oral toxicity of Samrannitra in accordance with OECD Guideline No. 408, emphasizing pharmacodynamic safety and systemic tolerance. Male and female Sprague-Dawley rats were randomly allocated into four groups, receiving 0 (control), 40, 200, or 400 mg/kg/day of Samrannitra via oral gavage for a duration of 90 days. A supplementary satellite cohort was monitored for 28 days following therapy to evaluate any delayed or reversible effects. Clinical symptoms, mortality rates, food and water consumption, and body weight were assessed on a weekly basis. Hematological and biochemical markers were evaluated at the conclusion of the trial. Principal organs, including central nervous system tissues, were weighed and subjected to histological examination. No adverse effects associated to therapy were found at any dosage. Growth trajectories, behavioral assessments, and physiological functions were consistently within standard parameters. The hematological and biochemical values were analogous to those of the control group. Histopathological assessment indicated no central nervous system or systemic organ damage resulting from the treatment. Samrannitra exhibited no signs of systemic or organ-specific toxicity, including central nervous system-related effects, at dosages of up to 400 mg/kg/day. The No Observed Adverse Effect Level (NOAEL) was determined to be 400 mg/kg/day. These findings offer critical safety pharmacology data to facilitate the clinical application of Samrannitra as a standardized herbal treatment for insomnia.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108438"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730682","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":"Simultaneous monitoring of heart rate, body temperature, and activity during postoperative recovery in group-housed telemetry-instrumented mice","authors":"Stephan Martin,&nbsp;Troy Velie,&nbsp;Justin Van Hee,&nbsp;Kimberly White,&nbsp;Julio Alvarez,&nbsp;Mike Girand","doi":"10.1016/j.vascn.2026.108480","DOIUrl":"10.1016/j.vascn.2026.108480","url":null,"abstract":"<div><div>Studying physiological parameters during post-surgical recovery is critical to gain insights into animal health. However, technological restrictions can pose practical challenges when conducting experiments with socially housed animals. To address this, we developed a sophisticated telemetry system, SoHo™, capable of simultaneously collecting electrocardiogram (ECG), body temperature, and locomotor activity in group-housed small animals. This study evaluates the efficacy of the SoHo™ telemetry platform to wirelessly monitor ECG, body temperature, and locomotor activity in group-housed mice. C57 female mice (22 weeks old; <em>N</em> = 5) were surgically instrumented (intraperitoneally) with the SoHo™ telemetry transmitters (model X01). The animals were group-housed under a standardized 12-h dark/light cycle. Synchronized ECG, core body temperature, gross locomotor activity, and video data were continuously collected during the post-operative recovery (12 days). Recorded heart rates (HR) following the surgical implantation were 607 ± 49 bpm (day) and 609 ± 42 bpm (night). The time course of the HR during the recovery period shows a decrease in HR of 13% (day) and 7% (night) on day 12. Telemetric core body temperature readings after surgery were 36.2 ± 0.18 °C (day) and 36.3 ± 0.22 °C (night) and increased by 0.6% (day) and 1.2% (night) at the end of the recovery time. Average gross locomotor activity (calculated as counts per minute; cpm) was 2659 ± 913 cpm (day) and 2232 ± 646 cpm (night) post-surgery. Daily activity decreased by 14% (day) and nightly activity increased by 72% (night) on day 12. The data demonstrates that SoHo™ telemetry is a powerful platform for evaluating ECG, temperature, and activity variations in socially housed mice. Further experiments, such as different pain treatments and group housing approaches, are needed to gain more details into post-operative recovery in socially housed mice.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108480"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730646","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":"Temperature-induced alterations in red blood cell osmotic resistance and aquaporin activity","authors":"Ipek S. Firat,&nbsp;Gerhard M. Artmann,&nbsp;Oliver H. Weiergräber,&nbsp;Samar Damiati,&nbsp;Özgür Alacayir,&nbsp;Aysegül Temiz Artmann","doi":"10.1016/j.vascn.2026.108490","DOIUrl":"10.1016/j.vascn.2026.108490","url":null,"abstract":"<div><div>Aquaporins (AQPs) are key players in regulating water movement across cell membranes and are increasingly recognized as potential drug targets, especially in conditions involving fluid imbalance. In this study, we explored whether AQPs and red blood cell (RBC) membranes undergo sudden changes at specific temperatures, a concept inspired by the Interfacial Water Quantum Transition (IWQ) model, which proposes that proteins switch functionally at critical temperatures. RBC samples of human and chicken RBCs were assessed in osmotic fragility (OF) studies using the classical method and scanned over a physiologically relevant temperature range of 24–50 °C. Mean corpuscular fragility (MCF₅₀) was determined applying a logistic curve fitting, and temperature effects were evaluated via Spearman's rank correlation. Human RBCs showed strong inverse correlations between temperature and MCF₅₀ in whole blood (ρ = −0.983), washed cells (ρ = −0.867), and AQP-inhibited cells (ρ = −0.983) (inhibition with 40 μM HgCl₂). Plasma removal increased the osmotic fragility, highlighting a protective role of extracellular plasma proteins. A distinct temperature transition at 36.0 ± 0.4 °C eliminated group differences, suggesting a temperature-induced switch in AQP functionality or membrane structure. At a transition temperature of T_C = 36.0 ± 0.4 °C, any differences between the groups were no longer identifiable, indicating a switch mechanism in AQP functionality or membrane structure at T_C. The transition was peak-like with a temperature range of ±0.4 °C around the critical value T_C. Chicken RBCs showed a similar, apparently species-specific transition at TC = 41.0 ± 0.5 °C. Interestingly, the Tc values of these two species corresponded to their respective body core temperature. These results suggest sharp, temperature-dependent transitions in membrane function that may be related to a switching of the AQP activity and water-protein interactions, as suggested by the IWQ model. The investigation of these transitions could contribute to the development of drugs targeting membrane proteins (AQPs), improve treatment strategies for fever or hypothermia, and long-term storage strategies in cryopreservation for maintaining the quality of red blood cells (RBCs) used for clinical purposes.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108490"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730651","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":"Cage divider as an alternative to single housing in studies","authors":"Hanne Daems,&nbsp;Renske Coopmans","doi":"10.1016/j.vascn.2026.108513","DOIUrl":"10.1016/j.vascn.2026.108513","url":null,"abstract":"<div><div>The EU Directive 2010/63/EU emphasizes that social species, such as rats, should be socially housed in stable groups of compatible individuals unless there's a strong justification otherwise. Any decision to individually house rats must be justified, considering the impact on the animal's welfare and potential effects on scientific results. The EU directive mandates that single housing should be limited to the shortest possible period and visual, auditory and olfactory contact should be maintained.</div><div>To assess the impact of an in-house modified transparent divider with large openings for interaction, the outcomes of removing the cagemate with inserting the divider were compared, relative to group housed conditions. In the second part of the study, the approach was reversed by reintroducing the cagemate into the original cage after 24 h of isolation or by removing the divider from the cage.</div><div>In this study, 4 male rats were included equipped with a telemetric device, which were group housed with a non-implanted cagemate. The telemetry apparatus facilitates continuous monitoring of cardiac hemodynamic parameters, including heart rate (HR), mean blood pressure (MBP), body temperature (BT), and locomotor activity.</div><div>When the cagemates were removed, an increase in HR, MBP and activity was noted. In contrast, when the divider was placed, minimal changes in HR, MBP and activity were observed. Only a slight decrease in BT was noted, likely due to less body heat of their cagemate.</div><div>After placing the cagemates back, aggression was immediately observed in 2 out of 4 cages, accompanied by clear increases in HR, MBP, activity and BT for 180 min, and again during the night period. Conversely, when the divider was removed, there were no signs of aggression, nor any changes in HR, MBP, or BT. The only notable difference was a slight increase in activity, as all animals began exploring the part of the cage they hadn't accessed for 24 h.</div><div>These findings suggest that group housing with a divider results in fewer stress indicators compared to single housing, during cardiovascular safety studies in which group housing is not applicable.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108513"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730716","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":"In vitro vascular ion channel screening for drug-induced vascular liability","authors":"Hannah G. Jennings Masters,&nbsp;Magali Ann Maizieres,&nbsp;Kim Rockley,&nbsp;Ruth Roberts,&nbsp;Michael Morton","doi":"10.1016/j.vascn.2026.108508","DOIUrl":"10.1016/j.vascn.2026.108508","url":null,"abstract":"<div><div>The control of blood pressure is complex, but vascular ion channels play a key role (Tykocki et al., 2017). Estimates suggest 20–30% of commonly prescribed medications can influence blood pressure; here we investigate the role of vascular ion channels in drug-induced blood pressure changes, offering the potential to establish a screen early in drug discovery.</div><div>Sixteen marketed drugs known to affect blood pressure were tested against a panel of vascular ion channels: Kv1.1, Kv2.1, Kv7.2/7.3, Kv7.3/7.5, CaV1.2, CaV3.2, Kir2.1. Drugs were tested in vitro by automated patch clamp (QPatch II, Sophion Biosciences). Drugs known to reduce blood pressure, like nifedipine and verapamil, showed activity at several ion channels, most notable vascular calcium channels. Clozapine which also changes blood pressure, showed activity at multiple vascular ion channels including Kv2.1 and Kv7.3/7.5 with IC50s of 13.0 and 42.1 suggesting a possible mechanism of action for clozapine. The data suggest that activity at ion channels is a potential mechanism for the effect of drugs on blood pressure. Thus, when testing unknown compounds as part of a screening strategy, activities at any of these ion channels may implicate possible effect on blood pressure. However, a negative result at these vascular ion channels cannot rule out the potential for effects on blood pressure by non-ion channel mediated mechanisms. Future work will build the panel by examining other relevant ion channels such as the KATP and K2P channels.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108508"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730086","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":"Development and characterization of non-human primate iPSC-derived ventricular cardiomyocytes for high-throughput safety pharmacology screening","authors":"Ilse Veldkamp-Baak,&nbsp;Pieter van Loenen,&nbsp;Silke Schwengberg,&nbsp;Mariana Argenziano","doi":"10.1016/j.vascn.2026.108494","DOIUrl":"10.1016/j.vascn.2026.108494","url":null,"abstract":"<div><div>Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are widely used in nonclinical cardiac safety studies; however, interspecies translation remains a limitation in early-stage pharmacology and toxicology. Non-human primates (NHPs) are physiologically closer to humans and are often used as secondary species for in vivo studies, providing translational insights. However, there are limited in vitro models using NHP-derived cardiomyocytes. We aimed to establish and characterize an <em>in vitro</em> platform of NHP iPSC-derived ventricular cardiomyocytes (vCardiomyocytes) suitable for cardiac safety pharmacology applications, focusing on consistency, functional responsiveness, and pharmacodynamic sensitivity to known reference compounds. iPSCs from cynomolgus macaques were differentiated into vCardiomyocytes at a large scale in stirred tank bioreactors and characterized across multiple production batches. Marker expression (cTnT, α-actinin, MLC2V) was quantified via immunocytochemistry and flow cytometry. Electrophysiological activity was assessed using multiwell microelectrode arrays (MEA). Field potential duration (FPD) variability (intra-well and inter-well) was evaluated. Functional responses were tested using reference cardioactive compounds. Complementary calcium flux and impedance-based contractility assays are ongoing to provide a broader assessment of functional integrity. The NHP vCardiomyocyte cultures showed &gt;93% cTnT-positive cells post-thaw with consistent co-expression of α-actinin and MLC2V. Spontaneous contractile activity emerged by day 3 post-thaw and persisted for at least 10 days with beat rates ranging from 28 ± 4.8 bpm and FPD of 528 ± 57.8 ms. MEA analysis revealed intra-well FPD variability of 2.19% and inter-well variability of 2.98%, indicating strong functional consistency across batches. Upon compound exposure, cells exhibited expected safety pharmacology phenotypes: arrhythmias with 10 nM dofetilide, beat arrest with 300 nM nifedipine, and &gt;20% beat rate increase with 100 nM isoproterenol. Calcium handling dynamics and contractility measurements are underway to confirm pharmacological sensitivity and functionality. NHP iPSC-derived vCardiomyocytes differentiated at large-scale display high structural fidelity, reproducible electrophysiological behavior, and pharmacodynamic responses aligned with known cardiac safety profiles. These cells represent a translationally relevant model for integrated assessment of proarrhythmic risk and other cardiac liabilities, supporting their application in nonclinical safety pharmacology workflows.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108494"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730256","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}