Journal of pharmacological and toxicological methods最新文献

{"title":"In silico predictions of drug-induced changes on cardiac contractility in human ventricular cardiomyocytes","authors":"Cristian Trovato ,&nbsp;Elisa Passini ,&nbsp;Maxx Holmes ,&nbsp;Stefano Longobardi ,&nbsp;Kylie A. Beattie ,&nbsp;Eric I. Rossman ,&nbsp;Blanca Rodriguez","doi":"10.1016/j.vascn.2025.107679","DOIUrl":"10.1016/j.vascn.2025.107679","url":null,"abstract":"<div><div>Drug-induced changes in cardiac contractility can lead to cardiotoxicity, a major cause of drug development discontinuation. Current routine <em>in vitro</em> strategies to assess changes in contractility for both small and large molecules are limited to low/medium throughput assays, not always correctly translating in clinical trials. The goal of this study is to present a computational framework for predicting drug-induced changes in human cardiac contractility and to compare <em>in silico</em> predictions with <em>in vitro</em> measurements of sarcomere shortening from adult human primary cardiomyocytes and clinical data available in the literature. In addition to simulating the behavior of pure specific ion channels blockers, a set of 28 reference compounds was simulated for this study. For each compound, half maximal inhibitory concentration (IC50) values were provided for the inhibition of four ionic currents: the fast Na + current, the L-type Ca2+ current (ICaL), the rapid and slow K+ currents. Wide ranges of drug concentrations, from 0.1× up to 100× of the free therapeutic plasma concentrations, were simulated in an experimentally-calibrated population of more than 300 human ventricular electromechanical <em>in silico</em> models constituting a healthy control population, with different ion channel expressions represented to account for cell-to-cell variability. Fifteen electrophysiological and contractility biomarkers were computed and analyzed. Active tension (AT) peak was identified as the most informative one and used to compute AT IC50. Pure ICaL blockers showed a strong reduction of AT peak and max/min AT velocity, whereas minor changes were observed for AT time-to-peak and AT relaxation. <em>In silico</em> predictions were qualitatively in agreement with <em>in vitro</em> recordings from adult human primary cardiomyocytes from literature for 27 out of 28 compounds. Cisapride was the only exception, showing no effects on simulated AT, whereas a negative inotropic effect was observed <em>in vitro</em>, despite there is no evidence of cisapride affecting contractility in clinical studies. Our study highlights the potential of <em>in silico</em> drug trials for early detection of drug-induced changes on cardiac contractility as a promising tool for use during drug development.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107679"},"PeriodicalIF":1.3,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927675","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":"State-of-the-art in secondary pharmacology and its impact on the safety of new medicines","authors":"Friedemann Schmidt ,&nbsp;Stephen Jenkinson ,&nbsp;Lyn Rosenbrier-Ribeiro ,&nbsp;Vicencia Toledo Sales ,&nbsp;Yoav Timsit ,&nbsp;Scott Mittelstadt ,&nbsp;Annie Delanois ,&nbsp;Jean-Pierre Valentin ,&nbsp;Richard Brennan","doi":"10.1016/j.vascn.2025.107680","DOIUrl":"10.1016/j.vascn.2025.107680","url":null,"abstract":"<div><div>Adverse effects of drugs often result from unexpected drug-off-target interactions triggering secondary pharmacology (2P). Consequently, 2P profiling of new chemical entities across a range of protein targets is a routine task in pharmaceutical R&amp;D. Particularly targets with known association to safety adverse events (<em>e.g.</em>, GPCRs, ion channels, transporters, protein kinases and other enzymes) are considered. However, there is significant variability across pharmaceutical industry in the composition of target panels screened, assay technologies implemented, and in the timing and scheduling of 2P screening efforts. As part of the IQ DruSafe consortium, a 2P working group was established, comprising representatives from 18 companies, to conduct a review of 2P strategies and share experiences, to recommend best practices and future application for the industry. Member companies contributed proprietary data across parameters such as target panel composition, position in the R&amp;D process, sourcing of the panels, the assay types and technology used. Performance and outcome, including total number of compounds run, concentrations tested, and hit rates, were also recorded. A detailed database capturing an overview of the strategic, scientific, and operational aspects of 2P screening has been created accordingly. This database currently holds 3729 records detailing on assays covering 747 distinct biological targets that are assessed for 2P. Conclusions will be presented highlighting some key trends that should be considered for future evolution of 2P practices across the industry. The working group identified a strong influence of previous published work on 2P panel composition by Bowes et al. 2012 and Lynch et al., 2017 with high prevalence of the targets they propose applied across the industry, but also a set of uncommon targets has been identified that should be considered from a safety perspective. Furthermore, given the recent broadening in pharmaceutical targets by precision medicine approaches, extending the coverage of 2P previously under-represented target classes such as protein kinases, enzyme families and epigenetic targets appears to be important.</div><div>Bowes et al. 2012, Bowes, J., Brown, A. J., Hamon, J., Jarolimek, W., Sridhar, A., Waldron, G., &amp; Whitebread, S. (2012). Reducing safety-related drug attrition: The use of <em>in vitro</em> pharmacological profiling. Nature Reviews: Drug Discovery, 11(12), 909–922.</div><div>Lynch, J. J., Vleet, T. R. V., Mittelstadt, S. W., &amp; Blomme, A. G. (2017). Potential functional and pathological side effects related to off-target pharmacological activity. Journal of Pharmacological and Toxicological Methods, 87, 108–126.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107680"},"PeriodicalIF":1.3,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927676","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":"JET, but not ‘snapshot’, analysis is sensitive enough to detect effects at critical concentrations of ondansetron and moxifloxacin in dog","authors":"Derek D. Best ,&nbsp;Matthew M. Abernathy ,&nbsp;Ken Kearney ,&nbsp;Derek J. Leishman","doi":"10.1016/j.vascn.2025.107605","DOIUrl":"10.1016/j.vascn.2025.107605","url":null,"abstract":"<div><div>Jacketed external telemetry (JET) is an important technology allowing access to continuous ambulatory electrocardiogram data in settings in addition to standalone cardiovascular safety pharmacology studies such as GLP toxicology studies. In developing oncology therapeutics under ICH S9 it may also be the case that QTc data collected during toxicology studies is the only in vivo data available to support an integrated risk assessment under the new ICH E14 Q6.1. The current analyses compared JET in freely moving dogs with more conventional ‘snapshot’ collections in restrained, recumbent animals for the same group of 24 beagle dogs (12 M, 12F). The treatments tested were vehicle, ondansetron (3 and 10 mg/kg), and moxifloxacin (30 mg/kg). A parallel study design (3 animals per sex per group) was used in the comparison of JET and ‘snapshot’ collection. In a subsequent phase of the study the 12 male dogs were rotated through the other treatments to create a Latin square crossover analysis. The higher dose of ondansetron achieved Cmax exposures consistent with the clinical critical concentration associated with a 10 ms QTc prolongation. The tested dose of moxifloxacin was the same as that tested in previous studies and achieved a Cmax concentration ~ 3-fold the critical concentration associated with a 10 ms QTc prolongation and had sustained exposure throughout the 24 h telemetry collection period. No treatment related QTc effects were detected using ‘snapshot’ data collection. The small heart rate increase following moxifloxacin detected using JET was not detected in the ‘snapshot’ analysis. Statistically significant, treatment-related QTc prolongation was detected for both doses of ondansetron and moxifloxacin in the parallel and the cross-over JET data analyses. The study-specific least significant difference was 7.1 ms and 2.6 ms for these analyses, respectively. JET data collection and analysis in a parallel toxicology-like design proved sensitive enough to support ICH E14 Q6.1 integrated risk assessment. Conventional ‘snapshot’ analysis was not sufficiently sensitive to detect a treatment-related effect even for modest multiples of the human critical concentrations.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107605"},"PeriodicalIF":1.3,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927792","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":"Ciprofloxacin does not accelerate aortic stiffness or aortic dilation in healthy, hypertensive and diseased mouse models","authors":"Callan Dwaine Wesley,&nbsp;Cedric H.G. Neutel,&nbsp;Dustin Kruger,&nbsp;Celine Civati,&nbsp;Guido R.Y. De Meyer,&nbsp;Wim Martinet,&nbsp;Pieter-Jan Guns","doi":"10.1016/j.vascn.2025.107670","DOIUrl":"10.1016/j.vascn.2025.107670","url":null,"abstract":"<div><div>Fluoroquinolones such as ciprofloxacin have been associated with increased risk of aneurysms and aortic dissections. This study investigated whether ciprofloxacin induced vascular remodelling leads to arterial stiffness in mice. Additionally, ciprofloxacin was evaluated in disease models, <em>i.e.</em> L-NAME-induced hypertensive mice with arterial stiffness and Fbn1C1039G+/− mice mimicking Marfan syndrome with elastin breaks and high arterial stiffness. 12-week-old C57BL/6 J and Fbn1C1039G+/− male mice were administered ciprofloxacin (100 mg/kg/day <em>via</em> drinking water) or vehicle for two periods of two weeks. Arterial stiffness and aortic diameter were measured both <em>in vivo</em> (pulse wave velocity (PWV) by ultrasound imaging, week 0,2,6 and 14) as well as <em>ex vivo</em> (Peterson's Modulus (Ep), week 14). Clinically relevant ciprofloxacin plasma concentrations (2.3 ± 0.4 μg/ml) were confirmed. However, ciprofloxacin treatment did not exacerbate arterial stiffness <em>in vivo</em> in wild type (WT) (PWV: 2.4 ± 0.1 <em>vs</em> 2.4 ± 0.2 mm/s), hypertensive (PWV: 3.5 ± 0.4 <em>vs</em> 3.5 ± 0.2 mm/s) or Marfan mice (PWV: 4.2 ± 0.3 <em>vs</em> 4.1 ± 0.2 mm/s). Furthermore, this was confirmed <em>ex vivo</em> when ciprofloxacin treatment did not affect arterial stiffness in WT (Ep: 296 ± 7 <em>vs</em> 295 ± 10 mmHg), hypertensive (Ep: 375 ± 11 <em>vs</em> 389 ± 9 mmHg) or Marfan mice (Ep: 595 ± 77 <em>vs</em> 569 ± 63 mmHg) when represented as control <em>vs</em> ciprofloxacin respectively. Differences in <em>in vivo</em> diameters were not reported after ciprofloxacin treatment within WT (D: 1.4 ± 0.03 <em>vs</em> 1.4 ± 0.03 mm), hypertensive (D: 1.3 ± 0.03 <em>vs</em> 1.2 ± 0.04 mm), or Marfan mice (D: 1,9 ± 0.20 <em>vs</em> 1,8 ± 0.03 mm) when represented as control <em>vs</em> ciprofloxacin respectively. The findings indicate that no adverse effects on arterial stiffness and aortic dilation were observed after ciprofloxacin treatment, which contrasts previous reports in patients and mice.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107670"},"PeriodicalIF":1.3,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927799","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":"Building a robust CNS screening strategy: Integrating in vivo and in vitro assays to better predict CNS risk","authors":"David A. Connor,&nbsp;Matthew Cato,&nbsp;XueJun Wu,&nbsp;Shannon Tringola,&nbsp;Eric I. Rossman,&nbsp;Ajeesh K. Cherian","doi":"10.1016/j.vascn.2025.107662","DOIUrl":"10.1016/j.vascn.2025.107662","url":null,"abstract":"<div><div>Attrition within clinical development due to central nervous system (CNS)-related adverse events continues to highlight the need to refine CNS screening strategies. The primary preclinical method to assess CNS risk is the Irwin/Functional Observation Battery (FOB). However, these tests are significantly limited by subjective assessment and temporal resolution. Automated neurobehavioral assessment, such as the Home Cage Analyzer (HCA), can overcome these issues. In addition, in vitro methods, such as microelectrode array (MEA), provide greater throughput and improved translation when using human iPSCs. Ideally, the combination of neurobehavioral and electrophysiological endpoints would converge to provide a clearer picture of CNS perturbance. Critically, a robust CNS screening strategy hinges upon understanding the limitations of such methods and identifying potential gaps in sensitivity between them. This study aimed to validate the Home Cage Analyzer (HCA) and test for convergence or difference in sensitivity between in vivo and in vitro screening using potent CNS active compounds from two distinct drug classes: psychostimulants and sedatives. For HCA experiments, eight male Wistar Han rats (Charles River) were implanted with temperature sensitive RFID transponders (Biomark USA) and pair housed in individually ventilated cages (Tecniplast). Rats were maintained under a 12-h light/dark cycle and habituated to the testing room and HCA rack for 1 week prior to initiation of dosing. Using a crossover Latin-square design, each rat received a single dose of d-amphetamine (0.25, 1, or 3 mg/kg), diazepam (0.5, 1.5, 5 mg/kg), or saline/vehicle. For MEA experiments, hiPSC-derived cortical neuron/astrocyte network (Fujifilm CDI) or rat cortex (E18.5; QBM Biosciences) was cultured in a Maestro Multi-electrode Array System (Axion BioSystems; 48-well plate). l-amphetamine in saline (0.12, 0.37, 1.11, 3.33, 10, 30 μM) or diazepam in DMSO (0.1, 0.3, 1, 3, 10 μM). As expected, we found a dose-dependent increase in activity after treatment with amphetamine in the HCA. Conversely, rats treated with diazepam showed decreased activity levels within the HCA. In line with neurobehavioral effects, diazepam caused a significant decrease in spiking and burst frequency in the MEA, resulting in a well-defined sedative phenotype. Surprisingly, amphetamine application in iPSCs exhibited no phenotypic response within the MEA, including no change in burst rate. Importantly, this lack of an effect in the MEA could be interpreted as a “clean” result, thus representing a missed hazard. In contrast, diazepam demonstrated consistent effects in the HCA and MEA. Together, these results indicate complementarity between in vitro MEA assay and in vivo automated HCA when screening sedatives while revealing a gap between these assays for amphetamine. Thus, a multi-modal approach is critical to a robust CNS screening strategy.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107662"},"PeriodicalIF":1.3,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927863","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 head twitch response in male C57BL/6J mice as a measure of 5-HT2a receptor activation","authors":"Ella Richardson,&nbsp;Shamila Griffiths,&nbsp;Sharon Rowton,&nbsp;Christopher Smith","doi":"10.1016/j.vascn.2025.107622","DOIUrl":"10.1016/j.vascn.2025.107622","url":null,"abstract":"<div><div>The head twitch response (HTR) is a rhythmic paroxysmal rotational head movement that occurs in mice following 5-HT_2A receptor activation. This test may be used to evaluate the potential for novel drugs to activate the 5-HT_2A receptor, to produce a synergistic effect when co-administered with medications which elevate serotonin levels or antagonize hallucinogenic-like behavior for the purpose of treating aspects of brain disorders, including positive symptoms of Schizophrenia. The HTR was characterized in male C57BL/6J mice using psilocybin (0.5 to 2 mg/kg, IP), psilocin (0.3 to 1.0 mg/kg, IP), LSD (0.05 to 0.2 mg/kg, IP) and DOI (0.25 to 1.0 mg/kg, IP). Immediately following administration, mice were placed into observation chambers and observed continuously for 60 min. The number of head twitches were recorded in 10-min bins. Head twitch was observed in all treatment groups and varied in terms of frequency and time of peak effect. DOI produced an inverted ‘<em>U</em>'-shaped dose response, peaking between 21 and 30 min postdose, with a group total of 85 head twitches (0.5 mg/kg). Psilocybin and psilocin (the pharmacologically active agent of psilocybin) produced a dose-related response, peaking between 11 and 20 and 0 and 10 min postdose, respectively, with a group total of 70 (2 mg/kg) and 55 (1 mg/kg) head twitches, respectively. LSD produced a dose-related response, peaking between 11 and 20 min postdose, with a group total of 63 head twitches (0.2 mg/kg). For the purpose of characterizing attenuation of the HTR, clozapine (atypical antipsychotic) at 1.25 to 5.0 mg/kg, IP or haloperidol (typical antipsychotic) at 0.15 to 0.6 mg/kg, IP, were administered to mice. Thirty minutes later, the hallucinogen DOI (0.5 mg/kg, IP) was administered to induce the HTR. Animals were observed continuously for 30 min from 10 min post-DOI administration. Clozapine (1.25 mg/kg) and haloperidol (0.6 mg/kg) attenuated HTR by 68 and 64 %, respectively, when compared to DOI alone. Clozapine (2.5 and 5 mg/kg) attenuated the HTR, however the animals were highly sedated. These findings validated the HTR test in male C57BL/6J mice.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107622"},"PeriodicalIF":1.3,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927504","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":"Assessment of abuse liability and respiratory effects of mitragynine and 7-hydroxymitragynine in rats","authors":"Julio D. Zuarth Gonzalez ,&nbsp;Ragsdale K. Alexandria ,&nbsp;Sushobhan Mukhopadyay ,&nbsp;Christopher R. McCurdy ,&nbsp;Lance R. McMahon ,&nbsp;Jenny L. Wilkerson","doi":"10.1016/j.vascn.2025.107624","DOIUrl":"10.1016/j.vascn.2025.107624","url":null,"abstract":"<div><div>Kratom (Mitragyna speciosa) is a natural product ubiquitously available in most US states with little, if any, regulation. Many use kratom to self-treat pain and opioid dependence, yet scientific evidence is lagging. Between 2011 and 2017 there were 1807 kratom exposures reported to poison control centers in the United States. Most of them occurred in the last two years (65 %) and resulted in a serious medical outcome (51.9 %). There is a pressing need to evaluate the safety of the active components found in the kratom plant, as the adverse effects reported to poison control centers could be attributed to any of the 40+ alkaloids identified within kratom. Among these alkaloids, mitragynine is the most prevalent in the plant, while 7-hydroxymitragynine, a minor alkaloid with a strong affinity for the mu-opioid receptor, has also been identified. This study aims to evaluate both the potential for abuse and the impact on respiratory parameters of mitragynine and 7-hydroxymitragynine.In jugular catheter-implanted male and female Sprague Dawley rats, intravenous (i.v.) self-administration for the opioid agonist remifentanil (1 μg/kg/infusion), was established and animals were trained on a multi-component paradigm in which different doses were available at each component. Once remifentanil response was stable, remifentanil was substituted for the test compounds (mitragynine, 7-hydroxymitragynine, and fentanyl). Respiratory experiments were conducted using a within-subjects design and included a 7-day drug washout period between sessions on a separate cohort of male and female rats. Respiratory frequency, tidal volume, and minute ventilation were measured, both pre and post-drug administration, using whole-body plethysmography in unrestrained animals. When mitragynine was substituted for remifentanil, mitragynine suppressed remifentanil-associated lever responses. However, either 7-hydroxymitragynine or fentanyl substitution maintained responding on the remifentanil-associated lever. This same profile was observed in respiration experiments in which mitragynine failed to produce respiratory depression, but 7-hydroxymitragynine resulted in respiratory depression similar to fentanyl. These results are in agreement with the in vitro and in vivo literature indicating mitragynine has low abuse potential and does not result in respiratory depression. However, 7-hydroxymitragynine may be subject to abuse liability and respiratory depression.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107624"},"PeriodicalIF":1.3,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927375","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":"Characterization of the electrocardiograms and hemodynamics of the HSD:Sprague-Dawley rat","authors":"Michael Stonerook,&nbsp;Bryce Visnick,&nbsp;Jeffrey Wallery","doi":"10.1016/j.vascn.2025.107612","DOIUrl":"10.1016/j.vascn.2025.107612","url":null,"abstract":"<div><div>Cardiovascular studies in rats are an invaluable aspect of the lead optimization and development process of pharmaceutical compounds. Electrocardiographic (ECG) and blood pressure (BP) data for male (<em>n</em> = 16) and female (<em>n</em> = 15) HSD:Sprague-Dawley rats were evaluated. Data was collected over eight 24-h periods over several months by means of implanted telemetric data collection (EMKA Technologies) and binned as 15-min interval means over each 24-h period. There were differences between the sexes for several ECG parameters; additionally, differences were seen within each sex group when compared to the grand means comparisons light and dark cycles. Heart rates (HR) were higher for females (HRDay = 431 ± 18 bpm, HRNight = 444 ± 11 bpm) as compared to males (HRDay = 401 ± 13 bpm, HRNight = 424 ± 12 bpm). There was a shorter PR interval in the females (PRDay 40 ± 1 ms, PRNight 39 ± 1 ms) compared to males (PRDay 44 ± 1 ms, PRNight 42 ± 1 ms) that were considered secondary to heart rate differences. All other ECG parameters had minimal differences between sexes and light and dark cycles. BP values (systolic, mean, diastolic) were always lower in females (mean BPDay = 110 ± 2 mmHg, mean BPNight = 109 ± 2 mmHg) compared to males (mean BPDay = 120 ± 3 mmHg, mean BPNight = 122 ± 2 mmHg) at all timepoints. Using power analysis, we determined that the highest sensitivity to detect changes in ECG or BP could be found if sexes were analyzed separately, as well as separate analysis of the light and dark cycles (conditions were <em>n</em> = 4/sex, power = 0.8, α = 0.05, and fixed effects were 5 % and 10 % difference of means). A review of QT interval in the rat found it was highly linearly related to RR interval, unlike large animal species, and that use of Bazett's correction formula was the most appropriate for normalization. In conclusion, it was determined that rat cardiovascular data required a different statistical approach compared to those of large species and that these approaches of separate sex and light cycle analyses could increase the power of the model to detect cardiovascular changes.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107612"},"PeriodicalIF":1.3,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927607","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":"Combining a systems immunology approach with in vitro data to evaluate drug-induced changes in the immune response after Influenza A Virus infection – A blueprint for the preclinical assessment of infection risks for biologics","authors":"Marcel Mohr ,&nbsp;Rami Lissilaa ,&nbsp;Andreas Hohlbaum ,&nbsp;Friedemann Schmidt","doi":"10.1016/j.vascn.2025.107682","DOIUrl":"10.1016/j.vascn.2025.107682","url":null,"abstract":"&lt;div&gt;&lt;div&gt;Biological modalities, such as antibody-based biologics, have revolutionized the treatment of many neoplastic and inflammatory diseases over the last years. Although being considered as less promiscuous to off-target interactions than synthetic immunosuppressive molecules, infections remain a key safety consideration. The immunosuppressive modulation of targeted immune system components allows infectious organisms a greater chance to spread or reactivate and cause disease in treated patients. Understanding how the modulated immune system combats infection is crucial to assess and predict potential complications of therapeutic biologics. The immune response to infections is a complex biological process, involving multiple cell types, pathways, and anatomical compartments, and is difficult to interpret solely at the level of &lt;em&gt;in vitro&lt;/em&gt; assays. Mathematical models and computer simulations have been widely used to study immune responses to infections, &lt;em&gt;e.g.&lt;/em&gt;, in case of Influenza A Virus (IAV). We used a mathematical model &lt;em&gt;[Lee et al. (2009), Journal of Virology]&lt;/em&gt; which represented published experimental findings on IAV infection to study the effect of a preclinical molecule on an IAV-specific immune response as compared to clinical or marketed competitor drugs with overlapping mechanism of action. The drugs are intended to inhibit autoreactive T cells in autoimmune diseases &lt;em&gt;via&lt;/em&gt; inhibition of co-stimulatory T cell receptors leading to a decreased proliferation of effector CD4+ and CD8+ T cells. For each drug, &lt;em&gt;in vitro&lt;/em&gt; data derived from mixed lymphocyte reaction assays were used to quantify the concentration-dependent inhibition of T cell proliferation and were integrated into the model. The drug-specific immune responses to IAV infection were simulated for two scenarios, &lt;em&gt;i.e.&lt;/em&gt;, primary IAV infection occurring during the treatment, and secondary IAV infection occurring during the treatment while IAV memory had already been established. Two &lt;em&gt;in silico&lt;/em&gt; biomarkers of the altered immune responses were generated and compared, &lt;em&gt;i.e.&lt;/em&gt;, concentration-dependent changes in the time to viral clearance and in the antibody levels up to 30 days. As a result, the drug-dependent modulation of immune responses to IAV infection were consistent for primary and secondary infection. Furthermore, modulation of the immune response by the preclinical candidate could be ranked between those of two already marketed drugs. This allowed us to exclude the burden of an overt risk associated with the preclinical candidate as compared to the competitors which have an established clinical safety profile. This &lt;em&gt;in silico&lt;/em&gt; study is generic since the systems immunology model is neither tailored to a specific modality, nor to the precise mode of action of the new biological or chemical entity. Instead, drug-specific actions on the immune system are represented by &lt;em&gt;in vitro&lt;/em&gt; data integrated into the mec","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107682"},"PeriodicalIF":1.3,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927611","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":"Assessment of myocyte ultrastructural changes in healthy, normal rats and in a rat model of mitral regurgitation: Implications for cardiovascular safety pharmacology","authors":"Pakit Boonpala,&nbsp;Anusak Kijtawornrat","doi":"10.1016/j.vascn.2025.107673","DOIUrl":"10.1016/j.vascn.2025.107673","url":null,"abstract":"<div><div>The ICH S7A and S7B are directed to assess blood pressure, heart rate, electrocardiogram, and arrhythmic risks. Recently, a new paradigm has shifted the focus to target the treatment of myocytes in heart failure patients. The ultrastructure of myocytes includes mitochondria, myofibrils, <em>etc.</em> Recent publications have shown that approximately 29 % of withdrawn drugs have been reported to exhibit mitochondrial toxicity. Until now, several animal models of human diseases have been applied to safety pharmacology studies which show a different safety profile from that observed in healthy animals. From literature, the mitral regurgitation (MR) is a volume overload model that shows evidence of ultrastructural changes. Therefore, this study aimed to define cardiac ultrastructural changes assessed by transmission electron microscope (TEM) in a rat model of MR compared with healthy, normal rat. MR was surgically induced in (<em>n</em> = 5) male, Sprague-Dawley (SD) rats, and sham procedures were performed on healthy male rats (n = 5). Echocardiography was performed to confirm the MR. Twelve weeks post-surgery, all rats were euthanized, and their hearts were used for TEM. This study revealed that the sham rats possessed a sarcomere that was well-structured and parallel. Their z-line boundary was obviously identified. In the MR rats, it was found that the z-line boundary was hazy, the sarcomere had destruction zones, and the sarcomere length was shortened (<em>P</em> &lt; 0.05). The morphology of the mitochondria was distinct in each of the three subpopulations of mitochondria. Each subpopulation (subsarcolemmal, interfibrillar, and perinuclear) mitochondrial morphology of the MR rats did not significantly differ from the sham rats. Dyadic junctional complex is situated at the 4 edges of the sarcomere. In the MR rats, the dyadic junction complex is disorganized and misplaced. In conclusion, TEM imaging provides strong evidence of the abnormalities that exist inside the cardiomyocytes. MR caused sarcomere shortening, mitochondrial morphology change, and sarcoplasmic reticulum remodelling that differed from those in healthy rats. These findings will provide a better understanding of the cardiac ultrastructural damages in a disease model and broaden the use of ultrastructure alteration to assess the potential drug cardiac adverse effects that might be observed in cardiovascular safety pharmacology studies.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107673"},"PeriodicalIF":1.3,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927669","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}