Journal of pharmacological and toxicological methods最新文献

{"title":"Temperature effect on hERG channel pharmacology measured using the qube automated patch clamp system","authors":"David Nagy ,&nbsp;Anders Lindqvist ,&nbsp;Mette Christensen ,&nbsp;Goran Mattson","doi":"10.1016/j.vascn.2025.107803","DOIUrl":"10.1016/j.vascn.2025.107803","url":null,"abstract":"<div><div>The human ether-à-go-go related gene (hERG) function is crucial for cardiac repolarization. Inhibition of the hERG channel can prolong cardiac action potentials, increasing the risk of ventricular arrhythmias, including torsade de pointes (TdP). Therefore, in vitro evaluations of compound effects on the hERG channel are routinely conducted in drug development to detect potential arrhythmic side-effects. Traditionally, these evaluations are performed at ambient temperatures. However, previous studies have shown that potency for certain compounds is underestimated when compared to tests at near-physiological temperatures. This study aims to highlight the importance of a temperature-controlled measuring environment for accurate evaluation of hERG channel blockers and to demonstrate the capabilities of the Qube automated patch clamp system in providing such an environment. We utilized the Qube 384 automated patch clamp system, equipped with a temperature control unit, to investigate the effects of temperature on concentration-response relationships for a panel of known hERG channel blockers. The Qube system allows for up to 384 parallel recordings at controlled temperatures ranging from 8 °C and above. Biophysical and pharmacological experiments were conducted to assess the impact of temperature on channel activation and inactivation kinetics, as well as compound potency. Our experiments showed that temperature control significantly influences hERG channel pharmacology. We observed an increased rate of activation, a leftward shift in steady-state activation, and a rightward shift in steady-state inactivation with rising temperatures. Pharmacological responses varied with different compounds; for instance, verapamil and quinidine potencies remained unchanged with temperature variations, while erythromycin, sotalol, E-4031, and cisapride exhibited pronounced leftward shifts in potency when temperature increased from 18 °C to 34 °C. The findings underscore the importance of temperature control in hERG channel evaluations. The Qube 384 automated patch clamp system, with its ability to regulate and standardize temperature at the measurement site, proves to be a reliable tool for routine compound testing under controlled temperature conditions. This study confirms that accounting for temperature is critical in accurately assessing the pharmacology of hERG channel blockers, thereby enhancing the predictive power of in vitro assays in drug development.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107803"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145094928","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 state-of-the-art method for high-throughput, automated locomotor activity testing in neuropharmacology, safety, and toxicology studies","authors":"Troy Velie ,&nbsp;Kathryn Nichols ,&nbsp;Kara Mendiola ,&nbsp;Kimberly White ,&nbsp;Kim Swearingen ,&nbsp;Francesco Mannara ,&nbsp;Anil Mehendale ,&nbsp;Guinevere Bell ,&nbsp;Mike Girand ,&nbsp;Chris Kolin ,&nbsp;Julio Alvarez ,&nbsp;Evelyne Cel ,&nbsp;Dilshan S. Harischandra ,&nbsp;Sarah A. Beck","doi":"10.1016/j.vascn.2025.107795","DOIUrl":"10.1016/j.vascn.2025.107795","url":null,"abstract":"<div><div>The quantification of spontaneous locomotor activity, stereotyped movements, anxiety-related behavior, and exploration parameters is pivotal in neuropharmacology, safety, and neurotoxicity studies to determine whether new chemical entities possess psychostimulant, sedative, or toxic effects. Such studies may involve large-scale assessments of locomotor activity in rodents under GLP compliance, which can be labor-intensive and time-consuming. To address these challenges, we have developed the VivaMARS system for automated, high-throughput (up to 30 subjects by session) locomotor activity testing in rodents. This study aims to validate the VivaMARS platform with two reference compounds: caffeine (CAF), which is known to increase rodent activity, and chlorpromazine (CPZ), which is known to decrease rodent activity. Experiments utilized healthy adult Sprague Dawley® male rats (7–8 weeks old, 250–300 g) and CD1 male mice (six weeks old, 25–30 g). Animals were divided in 5 groups (<em>N</em> = 4 each): saline, low-dose, or high-dose groups for two reference compounds. CAF (low 4 mg/kg and high 16 mg/kg for both mice and rats) and CPZ (low 3 mg/kg and high 10 mg/kg for both mice and rats) were administered intraperitoneally. Data was recorded for 60 min immediately after CAF administration and 30 min after CPZ administration. The parameters quantified were total activity, subdivided into activity with displacement (ActiD) and without displacement (ActiND), distance travelled, vertical activity, immobility time, and speed characterization. Data were acquired and analyzed using the GLP-compliant Ponemah software. Table 1 details the effects of CAF and CPZ on measured parameters in mice and rats. In brief, a dose-dependent effect on ActiD was observed in both species after CAF (increase) and CPZ (decrease) administration. CAF-treated rodents travelled longer and CPZ-treated shorter distances compared to saline. As expected, CAF reduced, and CPZ increased immobility time in both mice and rats. CAF increased vertical activity in both species, while CPZ reduced vertical activity. The average speed was higher in CAF-treated animals and lower in CPZ-treated animals. Our data demonstrates that VivaMARS is a powerful platform for acute locomotor activity testing in rodents. Further experiments are needed for a more comprehensive validation of the instrument.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107795"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095100","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":"Variability of manual patch clamp hERG data generated using standardized protocols and following ICH S7B Q&A 2.1 best practices","authors":"Claudia P. Alvarez Baron ,&nbsp;Jun Zhao ,&nbsp;Huimei Yu ,&nbsp;Ming Ren ,&nbsp;Nicolas Thiebaud ,&nbsp;Donglin Guo ,&nbsp;Ryan DePalma ,&nbsp;Mistry Sabyasachy ,&nbsp;Isra Tariq ,&nbsp;Md Shadiqur Rashid Roni ,&nbsp;Omnia A. Ismaiel ,&nbsp;Murali K. Matta ,&nbsp;Manni Mashaee ,&nbsp;Jose Vicente ,&nbsp;Lars Johannesen ,&nbsp;Jiansong Sheng ,&nbsp;Simon Hebeisen ,&nbsp;James Kramer ,&nbsp;Andrew Bruening-Wright ,&nbsp;Koji Nakano ,&nbsp;Wendy W. Wu","doi":"10.1016/j.vascn.2025.107801","DOIUrl":"10.1016/j.vascn.2025.107801","url":null,"abstract":"<div><div>The most common mechanism of drug-induced QT_C prolongation and the potentially fatal arrhythmia <em>Torsade de Pointes</em> is block of hERG channels. Accordingly, the hERG assay is used to assess cardiac safety of new drugs in support of first-in-human studies. The recently updated ICH E14 Q&amp;As 5.1 and 6.1 describe regulatory pathways to use hERG results obtained following best practice recommendations (ICH S7B Q&amp;A 2.1) to complement clinical QT_C data that otherwise may not be adequate and inform labeling. However, the impact of protocol standardization on variability of hERG data has not been assessed. This is a critical data gap in implementation of the E14/S7B Q&amp;As. This hERG dataset was collected as part of a HESI-coordinated international effort designed to generate cardiac ion channel data using physiologically relevant protocols when feasible and practical, following best practice recommendations in ICH S7B Q&amp;As. Datasets for other cardiac ion channels are presented in a companion abstract (Yu et al.). Five laboratories established drug block potencies (IC₅₀s) for 28 clinical drugs in a two-phase study using manual patch clamp. Concentration verification was done to assess drug losses for all laboratories and drugs. Meta-analysis was used to estimate overall variability in IC₅₀s. Phase 1 study showed that hERG IC₅₀s were similar for four laboratories and systematically higher for one laboratory. The source of the systematic difference could not be identified and was not attributed to different extent of drug loss, drug delivery method, cell lines, or recording quality. The systematic difference disappeared during phase 2. Blind repeat testing of two phase 1 drugs was done by each laboratory to understand within the laboratory reproducibility. The overall variability (or reproducibility) of the hERG assay was estimated accounting for potency differences among different drugs and laboratory-specific tendencies. This variability indicates the resolution limit of the hERG assay under best practices to distinguish two IC₅₀s as different. Systematic differences in hERG IC₅₀s can occur following protocol standardization and ICH S7B best practice recommendations. The regulatory framework for identifying hERG-positive molecules should account for hERG data variability and may additionally need to account for laboratory-specific differences.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107801"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095110","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 of a perforated patch clamp assay for studying IPSC-derived cardiomyocytes","authors":"Daniel R.P. Sauter,&nbsp;Arianna Toppi","doi":"10.1016/j.vascn.2025.107802","DOIUrl":"10.1016/j.vascn.2025.107802","url":null,"abstract":"<div><div>Ion channel profiling of drug candidates using heterologous cells is a robust de-risking strategy. However, this approach often neglects the intricate interactions of intracellular components, leading to a growing interest in more physiologically relevant models. The quality of iPSC-derived cardiomyocytes has steadily improved, making these cells widely used in drug discovery due to their close resemblance to native cardiomyocytes. Patch clamp recordings, particularly action potential investigations, are the preferred method for evaluating the electrophysiological properties of cardiomyocytes. In automated patch clamp (APC) recordings, the whole-cell (WC) configuration is typically employed, but this can hinder action potential measurements as cytoplasmic components are ‘washed out,’ altering channel activities and disrupting Ca2+ buffering systems. Consequently, recorded action potentials are very short, especially during early repolarization. This effect is further exacerbated when fluoride is used as a seal enhancer, as Ca2+ and F- readily precipitate as CaF2. In this study, we developed an assay utilizing perforated patch clamp on a high-throughput automated patch-clamp (APC) platform to record action potentials from human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) using nystatin as a pore-forming agent. The application of the intracellular voltage-gated sodium channel (VGSC) blocker QX314 via internal solution exchange resulted in complete inhibition of VGSC currents in cells recorded in WC configuration, but had minimal effect on cells recorded in the perforated configuration, confirming that cells did not spontaneously transition to the WC configuration. Action potentials were recorded from cells matured between 8 and 21 days <em>in vitro</em>. Depending on the maturation time, action potential duration at 30 % repolarization (APD30) values were between 3.5 and 1.6 times longer in perforated patch clamp recordings compared to cells recorded in the WC configuration. Voltage clamp recordings revealed that this change in APD was attributable to larger voltage-gated calcium channel (VGCC) currents. These findings These findings underscore the advantage of using perforated patch clamp, as it allows for more accurate electrophysiological measurements by preserving the cell's native intracellular environment.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107802"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095111","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 of an in vitro screening assay to predict acute CNS toxicity induced by antisense oligonucleotides and mechanistic exploration of its toxicity","authors":"Keisuke Yoshikawa ,&nbsp;Yosuke Kato ,&nbsp;Mai Takizawa ,&nbsp;Chinami Aruga ,&nbsp;Toshiki Kagawa ,&nbsp;Naoto Uchibayashi ,&nbsp;Nami Nagafuku ,&nbsp;Yuto Ishibashi ,&nbsp;Naoki Matsuda ,&nbsp;Ikuro Suzuki ,&nbsp;Naoki Inamura","doi":"10.1016/j.vascn.2025.107844","DOIUrl":"10.1016/j.vascn.2025.107844","url":null,"abstract":"<div><div>In the development of antisense oligonucleotides (ASOs) indicated for central neurological diseases, local administration to central nervous system (CNS) (e.g., intrathecal injection) is selected as a clinical route of administration. However, some ASOs injected to CNS induce acute CNS toxicity such as ataxic gait, decreased locomotor activity, and convulsions in <em>in vivo</em> non-clinical studies. Therefore, it is desirable to screen the potential toxicities in the early stage of drug discovery. In this study, we tried to develop an <em>in vitro</em> screening tool to predict the risk of acute CNS toxicity induced by ASOs. We also explored the mechanism of the toxicity using <em>in vitro</em> methods. First, we investigated the effects of ASOs on neuronal activity by Ca²⁺ oscillation (CaO) assays using rat primary cerebral cortical neurons. A total of 27 gapmer-type ASOs, which had been evaluated in mouse intracerebroventricular (ICV) single dose toxicity studies, were evaluated as test articles. The results showed that P-rate (oscillation peak numbers per one minute) was inhibited in an ASO in concentration-proportional manner and that IC50 values generally correlated with the intensity of toxicity in the mouse ICV studies. Using “IC50 = 17 μM” as a cutoff value, intolerable acute toxicity observed in the mouse ICV studies could be predicted with a high accuracy (sensitivity: 83 %, specificity: 100 %). Furthermore, multielectrode array (MEA) assays using human iPS cell-derived neurons were conducted for some ASOs to investigate the potential mechanism of the acute CNS toxicity. Two types of similarity analysis (principal component analysis and AI analysis) based on the MEA data including reference small molecules suggested that the toxic ASOs were similar to AMPA glutamate receptor antagonists. Indeed, this mechanism was confirmed in patch clamp assay using HEK293 cells expressing human AMPA receptor subunits, GluR2. In summary, we developed a useful <em>in vitro</em> assay to screen the potential risk of acute CNS toxicity induced by ASOs and revealed that one of the mechanisms of the acute toxicity may be AMPA glutamate receptor antagonism.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107844"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095262","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":"Refining cardiovascular liability - Bayesian analysis of safety pharmacology in vivo studies","authors":"Richard Virgen-Slane ,&nbsp;Dingzhou Li ,&nbsp;Todd A. Wisialowski","doi":"10.1016/j.vascn.2025.107784","DOIUrl":"10.1016/j.vascn.2025.107784","url":null,"abstract":"<div><div>The ICH E14/S7B Q&amp;As includes Best Practice Considerations for the In Vivo QT Studies. In particular, the nonclinical studies for QTc prolongation should demonstrate adequate sensitivity and power to detect a QTc prolongation effect with a similar magnitude observable by dedicated clinical QT studies i.e., ΔQTc +10 msec. This is in accordance with the 3R (reduce/refine/replace) principle which aims to minimize the number of animals used on studies. In order to use the nonclinical and clinical data for an integrated assessment as a substitute for a thorough QT/QTc (TQT) study, unequivocal evidence of low QTc risk needs to be established using nonclinical in vitro and in vivo studies as defined for a nonclinical double-negative. To this end, we have developed a species-specific Bayesian paradigm that makes use of data collected from historical safety pharmacology studies in dog or monkey to construct the prior of all CV/telemetry endpoints, including QTc, with respect to both the circadian rhythm and dose response. This model was inspired by a publication co-authored by FDA on the subject, and yet considers features of our internal cardiovascular telemetry studies. We have built this model into an R package containing a shiny app to facilitate the analysis by Safety Pharmacology study directors and scientists. For most of the case studies examined, the Bayesian analysis improved the precision of the treatment effect assessment (5 % to 20 % reduction in the least significant difference (LSD)). By randomly removing animals from the study, we demonstrated the Bayesian method's ability to restore the accuracy even with incomplete data. Also, the Bayesian method provides a natural probabilistic statement of the treatment effect to facilitate decision making. All procedures performed on animals were in accordance with regulations and established guidelines and were reviewed and approved by an Institutional Animal Care and Use Committee or through an ethical review process.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107784"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095279","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":"Safety Pharmacology: Achieving a quarter century milestone as a scientific discipline","authors":"Michael K. Pugsley ,&nbsp;Brett R. Winters ,&nbsp;Stephen D. Tichenor ,&nbsp;Simon Authier ,&nbsp;Yevgeniya E. Koshman ,&nbsp;Krystle G. Correll ,&nbsp;Michael J. Curtis","doi":"10.1016/j.vascn.2025.108391","DOIUrl":"10.1016/j.vascn.2025.108391","url":null,"abstract":"<div><div>This editorial prefaces the annual themed issue on those methods with application to safety pharmacology (SP) in the <em>Journal of Pharmacological and Toxicological Methods</em> (JPTM). Highlighted content is derived from the 2024 Safety Pharmacology Society (SPS) meeting held in San Diego, CA, USA. The meeting showcased 122 posters, many of which are reproduced as abstracts published in JPTM. The manuscripts predominantly reflect updates to core battery safety evaluation and data analysis methods and include areas of novel investigation within SP. The results from several surveys including an updated salary survey by the SPS, current industry practices on neurotoxicity by the ACT, SPS and STP and results on the need to revisit the ICH S7A guidance on safety and secondary pharmacology by EFPIA, JPMA, and PhRMA provide timely updates. Other manuscripts include in vitro assessment methods for sodium channel block using MEA arrays, and a comparison of the in vitro effects of positive control drugs on the hERG channel current using different testing procedures. In vivo cardiovascular manuscripts include an overview of surgical telemetry implantation methods, utility of automated blood sampling methods in SP studies, evaluation of the performance characteristics for common QTc data collection methods, and an evaluation of a clinically used wearable ECG device for use in SP studies. There is also an overview of the impact of respiratory SP and a comprehensive review and comparison of current practices regarding methodological approaches used to acquire CV data in repeat-dose non-rodent toxicology studies. The 21 years of consecutive themed issues on SP methods attends to the importance of methods evaluation and the contribution of SP to the process of methods evaluation.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 108391"},"PeriodicalIF":1.8,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144890531","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":"Evaluation of the Effects of cypermethrin Toxicity on Liver Mitochondria of Male Albino Rats","authors":"Venkataramanaiah Poli ,&nbsp;Srinivasulu Reddy Motireddy","doi":"10.1016/j.vascn.2025.108388","DOIUrl":"10.1016/j.vascn.2025.108388","url":null,"abstract":"<div><div>The purpose of this study is to look into the toxic effects of giving male albino rats cypermethrin (CPM). In order to achieve this, three groups of rats were formed. Every group received the same amount of saline, including the control group, while the experimental groups received 4.5 and 5.5 mg/kg body weight of cypermethrin. After that, the rats are sacrificed, and tissue samples are pooled. In all the treatments, rats were administered orally for an experimental period of 30 days. Every week, the body weight of every individual rat was recorded. In order to achieve this goal, indicators of toxicity such as body and organic weight, ATP and ATP/ADP ratio, antioxidant enzymes and oxidative stress, complex activity (complex I–IV), plasma biochemical parameters, and levels of pro-inflammatory cytokines are utilized. Rats injected with cypermethrin showed a substantial (<em>p</em> &lt; 0.05) drop in their ultimate weight and organ weight when compared to control rats. The administration of cypermethrin resulted in a significant (<em>p</em> &lt; 0.05) drop in adenosine triphosphate (ATP) and the ATP/ADP ratio, indicating the involvement of activity of liver mitochondria. The administration of cypermethrin oxidative stress biomarkers resulted in a substantial increase (<em>p</em> &lt; 0.05) in malondialdehyde (MDA), glutathione (GSH), reactive oxygen species (ROS), and protein carbonylated content (PCC). Nevertheless, there was a significant (<em>p</em> &lt; 0.05) drop in the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione S-transferase (GST) in the mitochondria of rat liver. In the present study, the complex activity (complex I–IV) and mitochondrial membrane potential (MMP) of rat liver mitochondria decreased significantly (<em>p</em> &lt; 0.05). The administration of Cypermethrin resulted in a significant (p &lt; 0.05) increase in plasma levels of triglycerides (TAG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin (BIL), total cholesterol (TC), high-density lipoprotein (HDL-C), low-density lipoprotein (LDL-C), and alkaline phosphatase (ALP). Pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-10) were observed to have higher plasma levels after cypermethrin injection (<em>p</em> &lt; 0.05) when compared to the control. The current study's findings demonstrated that rat liver tissue suffered mitochondrial damage as a result of cypermethrin intoxication.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 108388"},"PeriodicalIF":1.8,"publicationDate":"2025-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144739316","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":"Legacy hERG study data in contemporary integrated risk assessments – mitigating the risk of protocol and positive control article differences","authors":"Richard W. Daniels ,&nbsp;James W. Kramer ,&nbsp;Matthew M. Abernathy ,&nbsp;Derek D. Best ,&nbsp;Jessica C. Brimecombe ,&nbsp;Derek J. Leishman","doi":"10.1016/j.vascn.2025.108387","DOIUrl":"10.1016/j.vascn.2025.108387","url":null,"abstract":"<div><div>Testing new molecules against the hERG potassium channel has been a routine component of secondary pharmacology assessments for more than a quarter of a century. In 2005, the ICH S7B guidance required a GLP hERG study prior to first administration of new chemical entities to man. In 2022, the ICH S7B Q&amp;As introduced some ‘best practice’ recommendations for the hERG assessment. A required component of a QTc integrated risk assessment is an evaluation of how well the hERG assessment met the ‘best practice’ recommendations. For studies conducted under the 2005 ICH S7B guidance, it is unclear how to assess the differences between study protocols. Many sponsors had been conducting the GLP hERG assay in a manner which was largely compliant with these recommendations but differed subtly in the recording temperature, the voltage-clamp protocol employed and/or in the positive control used in the assay (‘Legacy Protocol’). This led many sponsors to consider repeating their earlier hERG assessment using the ‘best practice’ (‘Revised Protocol’) recommendations. The current analyses sought to examine whether the differences in practice warranted retesting the compounds or could dispel a concern that prior practice had been less sensitive.</div><div>The three key positive control articles, dofetilide, moxifloxacin and ondansetron were compared under the ‘Legacy Protocol’ and best practice ‘Revised Protocol’. Furthermore, two prior positive control articles, terfenadine and cisapride, were tested under both protocols. Terfenadine data under the ‘Legacy Protocol’ were available from separate studies (<em>n</em> = 6) dating from 2003 to 2025. Terfenadine was also tested in four studies using the ‘Revised Protocol’ in 2023. In addition, the hERG inhibition data for 3627 single concentration positive control article tests with 60 nM terfenadine from studies conducted from 2002 to 2018 were collated.</div><div>There was no difference for the 3 positive control articles tested under the two different protocols for pooled hERG safety margin. Similarly, the pooled hERG IC₅₀ values for terfenadine under the two protocols were not different, although the variability in hERG potency was greater under the ‘Revised Protocol’. The inhibition of hERG current for 60 nM terfenadine in the 3627 historical examples was also consistent with predictions based on the hERG IC₅₀ and Hill slope data under the two protocols.</div><div>Based on the similarity between results collected under ‘Legacy’ and ‘Revised’ protocols we conclude that if the ‘Legacy Protocol’ has been used and the positive control examined was 60 nM terfenadine, there appears to be no need to repeat the hERG assessment.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 108387"},"PeriodicalIF":1.3,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144669182","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 retrospective analysis of the concordance between cardiovascular effects in telemetry studies, toxicity studies, and early clinical trials","authors":"Andrea Greiter-Wilke ,&nbsp;Jan Attig ,&nbsp;Simon Bassett","doi":"10.1016/j.vascn.2025.108385","DOIUrl":"10.1016/j.vascn.2025.108385","url":null,"abstract":"<div><div>This work aimed to compare the ability of several ECG technologies used in toxicity studies to identify cardiovascular effects. A retrospective analysis of 41 drug development projects across the Roche portfolio from 2005 to 2023 was conducted to evaluate the concordance of snapshot ECG (SS-ECG) and jacket external telemetry (JET, with or without blood pressure assessment) to detect effects on heart rate, ECG, and blood pressure in toxicity studies, when compared to implanted telemetry as the gold standard. Overall, 132 non-clinical studies were investigated. For 24 projects, translation to phase 1 safety data under consideration of plasma exposures could be followed up. Company-internal strategic decisions on advancing certain molecules despite cardiovascular findings were subject to a risk-benefit assessment and not evaluated in this investigation.</div><div>SS-ECGs never detected changes in heart rate (HR) previously revealed in telemetry studies, whereas JET identified these in all telemetry-positive cases. JET (or M11 implants) detected 44 % and SS-ECGs identified 33 % of QTc increases in telemetry-positive molecules, indicating that stand-alone telemetry is still the most reliable in vivo study in identifying QTc-effects. No clear species differences between beagle dogs, Göttingen minipigs or cynomolgus monkeys were evident when comparing heart rate and QTc detection. HR increases were noted clinically in about 50 % of the non-clinical positive cases, whereas HR decreases were never detected in clinical trials. Non-clinical blood pressure decreases showed higher translatability to humans (at least 80 %) than blood pressure increases (29 %), with a high prevalence of dogs and minipigs showing a signal. Non-clinical QTc increases detected by JET and/or telemetry were confirmed in 3 clinical studies analyzed with concentration/QTc modeling, however in 7 clinical studies employing other methodologies to measure QT, the non-clinical QTc effects did not translate.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 108385"},"PeriodicalIF":1.3,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144669181","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}