Journal of pharmacological and toxicological methods最新文献

{"title":"Rodent models of anxiety and compulsion: When multiple endpoints add value","authors":"Justyna Glazar,&nbsp;Iain Porter,&nbsp;Victoria Ascough,&nbsp;Sharon Rowton","doi":"10.1016/j.vascn.2025.107842","DOIUrl":"10.1016/j.vascn.2025.107842","url":null,"abstract":"<div><div>Anxiety disorders are the most diagnosed mental illnesses and exist independently or as comorbidity with conditions such as autism spectrum disorder, major depressive disorder, and/or substance use disorder. The acute treatment of moderate to severe anxiety includes medications, such as benzodiazepines, whereas for long-term treatment and compulsive disorders, selective serotonin reuptake inhibitors (SSRIs) are often prescribed. The objective of this investigation was to demonstrate the advantages of conducting a battery of behavioral tests to characterize the anxiolytic, anxiogenic, and/or anti-compulsive properties of drugs. Animal anxiety models are based on the natural tendency of rodents to avoid a potentially dangerous situation (e.g., open, brightly lit, or novel environments). Animal models of compulsive-like behavior are based on natural, repetitive behaviors exhibited by rodents (e.g., digging). Within this investigation, assessments were conducted using the elevated plus maze (EPM), staircase, light/dark box, Nestlet shredding, and marble burying tests in male C57BL/6 J mice. Investigations were conducted following intraperitoneal administration of 10 mg/kg paroxetine, fluoxetine, or atropine; 6 mg/kg diazepam; 0.1 mg/kg WIN55,212–2; or 4 mg/kg yohimbine; these doses did not adversely affect locomotor activity. Results show how, by evaluating multiple endpoints, results can be interpreted in terms of compulsion, anxiety, and in some instances, impulsivity with a greater degree of confidence. For example, atropine decreased marble burying by 63 %, Nestlet shredding by 94 %, and time in the light zone by 63 % compared with controls, demonstrating that effects on marble burying and Nestlet shredding were not due to anti-compulsive effects or anxiolysis. These preliminary investigations support the requirement for conducting testing for multiple endpoints when characterizing the potential anxiety or compulsive effect of a novel drug. Multiple endpoints can be considered within the same animals and may be considered for inclusion within toxicology studies.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107842"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095380","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":"Standardizing high-throughput RNA extraction: Modifying commercial kits for improved purity, yield, and efficiency","authors":"Ruwini D. Rajapaksha,&nbsp;Catherine Brooks,&nbsp;Adriana Rascon,&nbsp;Adam Fadem,&nbsp;Ivy Nguyen,&nbsp;Philip J. Kuehl,&nbsp;John T. Farmer","doi":"10.1016/j.vascn.2025.108379","DOIUrl":"10.1016/j.vascn.2025.108379","url":null,"abstract":"<div><div>High-throughput extraction kits are widely use in clinical and preclinical settings to extract nucleic acids from biological samples due to their scalability, repeatability, and reduced labor. However, achieving high nucleic acid yield and purity while maintaining reliability and reproducibility remains challenging in molecular biology labs, partly due to lack of standardized guidelines for nucleic acid extractions and purification. Variations in extraction kits and extraction chemistries often lead to inconsistence results across experiments.</div><div>To address this variability, we modified manufacturers' extraction protocols by introducing additional chloroform and ethanol extraction steps. These modifications aimed to optimize RNA purity, yield, and extraction efficiency (EE) using Xeno Internal Positive Control (IPC) spiking as a benchmark. We evaluated the performance of three commercially available magnetic-bead-based RNA extraction kits across six naïve non-human primate (NHP) tissue types: brain, heart, kidney, liver, lung, and spleen, using the KingFisher™ Flex automated extraction platform.</div><div>The modified protocols demonstrated significant improvements in RNA purity, yield, and EE across the selected kits. These additional steps enhanced the suitability of extracted RNA for downstream applications, underscoring the importance of considering both kit performance and tissue characteristics in experimental design.</div><div>We further assessed the MagMAX™ <em>mir</em>Vana™ Total RNA Isolation Kit for its ability to remove interfering nucleic acids, such as plasmid DNA and single-stranded DNA (ssDNA), from adeno-associated viral (AAV) vector preparations. Specifically, we invesigated AAV serotype 8 (AAV8) due to its popularity in use for gene and cell therapies. The kit demonstrated high efficiency, removing ≥98 % of non-encapsidated genomes, plasmid DNA, and other impurities, yielding RNA suitable for downstream applications and removing contaminating DNA that would create a false-positive signal and result in over quantification.</div><div>These results emphasize the value of optimized, standardized protocols to improve reproducibility and reliability in RNA extraction workflows, with broad applications in molecular biology, gene therapy, and cell biology research.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 108379"},"PeriodicalIF":1.3,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144517504","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 a human wearable ECG device for continuous data collection in freely moving non-rodents","authors":"Christopher P. Regan,&nbsp;Alysia A. Chaves,&nbsp;Theodore Detwiler,&nbsp;Jude Ferraro,&nbsp;Shaun Gruver,&nbsp;Desiree Steve,&nbsp;David Lengel,&nbsp;Julia C. Hotek","doi":"10.1016/j.vascn.2025.108377","DOIUrl":"10.1016/j.vascn.2025.108377","url":null,"abstract":"<div><div>Wearable devices are commonly used in clinical diagnostic medicine/personal health, but their use to collect nonclinical endpoints is limited due species-specific design specifications, data accessibility limitations, and availability of nonclinical telemetry technologies. These create a challenge to adopt and deploy clinical innovation in nonclinical studies. To determine the feasibility of using a human ECG wearable device (WRB) in nonclinical studies, we compared heart rate and ECG intervals between the simultaneously collected ECG from the WRB and implanted telemetry (TEL) device in canine (<em>n</em> = 5) and non-human primate (NHP, <em>n</em> = 4). Continuous ECGs were collected simultaneously from both devices for 24 h prior and 24 h after oral administration of vehicle or Dofetilide (canine <em>n</em> = 5; 0.003, 0.010, 0.030 mg/kg and NHP <em>n</em> = 4; 0.03, 0.06, 0.12 mg/kg). Individual animal 15-min means were reviewed descriptively and via Bland-Altman (BA) analysis to determine bias and limits of agreement (LOA) between devices. Further, Dofetilide dependent QT changes were evaluated using time-matched QT interval comparisons and the results of QTci exposure-response modeling. Data were qualitatively consistent in magnitude and profile between the two measurement platforms. BA demonstrated that bias between devices was &lt;2 ms for HR, PR, and QRS for both species, 2 ms for canine QT, and 6.6 ms for NHP QT, with acceptable LOA for all endpoints. Mean Dofetilide-dependent QT changes over time were not different between devices and calculated Δ∆ QTci EC_+10ms were &lt; 2× between devices within species. Overall, these studies demonstrated the feasibility of using alternative devices and increasing options to collect in-cage ECG to collect nonclinical safety pharmacology and toxicology endpoints.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 108377"},"PeriodicalIF":1.3,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144340715","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 cellular waste to biomarkers; insights into past, present, and future methods to detect immune cell-derived extracellular vesicles using flow cytometry","authors":"Jennifer L. Zagrodnik ,&nbsp;Craig S. Moore","doi":"10.1016/j.vascn.2025.108376","DOIUrl":"10.1016/j.vascn.2025.108376","url":null,"abstract":"<div><div>Extracellular vesicles (EVs) often possess both ubiquitous and unique tetraspanin molecules that can help elucidate their cell-of-origin. Furthermore, the presence and/or absence of specific tetraspanins can be used to phenotype and identify specific subpopulations of EVs. In the context of immune-related disorders (<em>i.e.</em> multiple sclerosis, rheumatoid arthritis, and various cancers), specific immune cell-derived EVs are now being investigated in the context of biomarker exploration, identifying novel disease mechanisms, and monitoring therapeutic responses in patients. Flow cytometry (FCM) is a technique that uses the light scattering properties of cells and/or subcellular particles (<em>e.g.</em> EVs), while combining fluorescent signals that can detect the presence, absence, or abundance of surface and/or intracellular molecules. To date, however, using FCM to accurately quantify EV populations has been challenging due to their relatively small size and weak light scattering and fluorescence properties compared to intact cells. Historically, the application of calibration beads of known sizes, refractory indices, a violet-side scatter, and standardized methodologies have made positive contributions towards the accurate detection and quantification of EVs while also permitting exploration into their biological properties. This review provides a summary and perspective of current FCM methodologies that are used to assess immune cell-derived EVs within biological fluids and cell supernatants. While acknowledging past and current limitations, as well as the recent successes, improvements, and efficiencies of assays used in EV-related research, the field will inevitably continue to advance through the implementation of standards and guidelines to enhance discovery and reproducibility.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 108376"},"PeriodicalIF":1.3,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144522949","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 cardiac safety evaluation: Assessing drug interactions with the late Nav1.5 current","authors":"Eva Vermersch ,&nbsp;Véronique Ballet ,&nbsp;Agnès Jacquet ,&nbsp;Irène Mahfouz ,&nbsp;Jean-Marie Chambard ,&nbsp;Françoise Chesney ,&nbsp;Desirae Martin ,&nbsp;Janette Rodriguez ,&nbsp;Ky Truong ,&nbsp;Najah Abi-Gerges ,&nbsp;Ambroise Garry","doi":"10.1016/j.vascn.2025.107806","DOIUrl":"10.1016/j.vascn.2025.107806","url":null,"abstract":"<div><div>Late Na + current (INa,L) contributes to cardiac action potential (AP) and helps maintain Na + homeostasis. Contrary to the effect on Nav1.5, hERG and Cav1.2 channels, the effect of novel drugs on INa,L is not routinely studied in safety studies. Yet, INa,L inhibitors can counterbalance hERG blockade and be associated with anti-arrhythmic potential. Hence, evaluating the effects of drugs against Nav1.5, hERG and Cav1.2 channels provides valuable cardiotoxic insights, but does not fully predict changes in the electrophysiological and contractile properties of cardiomyocytes. To address this deficiency, we evaluated the effects of selective and non-selective INa,L inhibitors on cardiomyocyte function. Compounds known to inhibit INa,L (GS-967 specific for INa,L; ranolazine specific for both hERG and INa,L; loperamide which is a hERG, Nav1.5 and Cav1.2 inhibitor) and four preclinical compounds were tested for their effects on cardiac ion channels (peak Nav1.5, hERG, Cav1.2 and INa,L) with automated patch-clamp and multi-electrode array (MEA) in hiPSC-derived cardiomyocytes for electrophysiological properties, and contractility in human primary cardiomyocytes from consented donor hearts with MyoBLAZER™. Each compound was tested separately at multiple concentrations in the presence of ATX-II, a selective enhancer of INa,L. GS-967 and ranolazine reversed ATX-II-induced increases in contractility and field potential duration (FPD) in a concentration-dependent manner providing evidence of a functional INa,L in both hiPSC-derived cardiomyocytes and adult cardiomyocytes. Next, we evaluated the effects of four preclinical compounds. Two out of the four compounds showed similar behavior to GS-967 and ranolazine. For example, compound A inhibited ion channels (hERG, Nav1.5, Cav1.2 and INa,L with IC50 values of 7.9 mM, 12.4 mM, 0.8 mM, respectively) and reversed ATX-II changes on contractility and FPD with IC50 values of 0.99 mM and 2.3 mM, respectively. Here, we developed a protocol for assessing drug interactions with INaL on cardiomyocytes. This assay enhances our ability to predict cardiotoxicity potential and its incorporation into the traditional compound derisking strategy strengthens confidence in advancing molecules into clinical development.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107806"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145094936","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 sensitivity/specificity of statistical analysis strategies for detecting moxifloxacin-induced QTc prolongation","authors":"Lawrence M. Carey,&nbsp;David Holdsworth,&nbsp;Jared Slain,&nbsp;Jill Dalton","doi":"10.1016/j.vascn.2025.107782","DOIUrl":"10.1016/j.vascn.2025.107782","url":null,"abstract":"<div><div>In light of the implementation of the best practice guidelines as outlined in the ICH E14/S7B Q&amp;A's, several strategies for enhancing the sensitivity of statistical analyses to detect drug-induced QT/QTc prolongation have been proposed. This study aimed to assess the sensitivity and selectivity of 3 standard statistical approaches. Moxifloxacin was administered orally at doses of 30, 80 and 175 mg/kg to male cynomolgus monkeys (<em>n</em> = 4) according to a Latin square design. Telemetry endpoints were monitored from 2 h prior to dosing to 24 h postdose. Three analysis strategies were employed to aid in the detection of QTc prolongation. Data were grouped into 3 analysis segments (0–6, 6–18, 18–24 h postdose). Approaches 1 and 2 utilized pairwise comparisons at each hourly interval within these segments or the entire segment based on the significance of the interaction effect, while approach 3 analyzed data pooled within each super-interval. Approach 1 employed a repeated measures analysis of variance (RMANOVA) strategy. Statistical significance was considered to have been attained when the <em>p</em>-value of each comparison was &lt;0.05. Approach 2 utilized an RMANOVA analysis strategy examining the least squares mean differences between each treatment and control. Statistical significance was considered to have been obtained when these differences were greater than or equal to the least significant difference (LSD) for the study and the 95 % confidence intervals (CI) did not cross zero. Approach 3 utilized an analysis of variance approach with statistical significance considered attained when the mean difference between each treatment and control was greater than or equal to the LSD and the 95 % CI did not cross zero. In general, the 3 approaches performed similarly. Approach 3 demonstrated the highest sensitivity (most statistically significant differences identified) while approach 2 displayed the highest selectivity (fewest type I statistical errors). Approach 3 performed better than Approaches 1 and 2 in detecting QTc prolongation. Of note, only approach 3 identified every QTc change greater than 10 msec as significant. All 3 methods of analysis were adequate to detect moderate changes in QT/QTc, though differences in the sensitivity and selectivity were observed between analysis strategies.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107782"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095275","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":"Study to compare effects of a positive inotrope administered concurrently with a beta-blocker in ventricular Tachy-Paced dogs in mild heart failure","authors":"Sydney St Clair","doi":"10.1016/j.vascn.2025.107836","DOIUrl":"10.1016/j.vascn.2025.107836","url":null,"abstract":"<div><div>Dilated cardiomyopathy(DCM) is common in dogs and is characterized by dilation and impaired systolic function of the ventricles. Pimobendan, a positive inotrope, has been suggested to benefit DCM by reducing preload and afterload, providing positive inotropic support, reducing cardiac size and filling pressures. Beta-blockers are used to treat ventricular arrhythmias associated with DCM, although there is concern that decreased inotropy could be detrimental to ejection fraction (EF). Given need for multimodal treatments, combined use of pimobendan and beta-blockers could help to mediate the negative inotropy of beta-blockers. This pilot study aims to compare the echocardiographic EF of simultaneous administration of pimobendan and sotalol (beta-blocker) at veterinary dosages in a HF dog model. Seven beagle-dogs were induced into mild HF using a ventricular-tachy-pacing(VTP) model. Dogs received four different treatment regimens: control, pimobendan (0.2–0.3 mg/kg PO BID), sotalol (1-3 mg/kg PO BID), and a combination of pimobendan and sotalol for two-weeks each. Multiple methods of calculating EF via echocardiography were evaluated, including ejection fraction (Eft) and fractional shortening (FS) from M-mode and Simpson's method-of-discs (SimpsonsEF) measured from right parasternal long-axis view, and were recorded at baseline, day 3, 7, and 14. By day 14, echocardiographic data indicated administration of pimobendan resulted in a significant increase in EF compared to baseline (↑ Eft 19.3 ± 3 % ΔBL; ↑ FS 24.1 ± 3.9 %ΔBL; ↑ SimpsonsEF 28.9 %ΔBL). In contrast, sotalol showed a significant decrease in EF compared to baseline (↓ Eft 9.5 ± 6.8; ↓ FS 9.9 ± 8.2; ↓ Simpson's EF 19.1 ± 9.5). The combination therapy of pimobendan and sotalol showed a non-significant increase in EF from baseline (↑ Eft 5.1 ± 5.5; ↑ FS 6.5 ± 6.7; ↑ SimpsonsEF 1.53 ± 9.5), but the change was similar to that of the control group (↑ Eft 3.1 ± 4.4; ↑ FS 4.2 ± 5.3; ↑ SimpsonsEF 3.0 ± 7.2), indicating that the change could be due to day-to-day variation. The study demonstrates that concurrent administration of a positive inotrope with a beta-blocker does not decrease cardiac function, as evidenced by no significant decrease in EF. In dogs with DCM that may benefit from beta-blocker therapy, pimobendan may be added to mitigate negative inotropic effects. This combination of therapies may allow clinicians to optimize treatment in complicated cases, warranting further investigation.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107836"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095355","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 strategy to predict psychotic risks of drug candidates using auditory steady-state response","authors":"Daigo Homma,&nbsp;Yoshiaki Furuya,&nbsp;Takashi Yoshinaga","doi":"10.1016/j.vascn.2025.107839","DOIUrl":"10.1016/j.vascn.2025.107839","url":null,"abstract":"<div><div>It is difficult to predict the psychiatric adverse effects of drug candidates from preclinical observations especially when the mechanism of actions is novel. Recently, auditory steady-state response (ASSR) in gamma band frequency has been reported as a translational biomarker for psychotic disorders and is thought to reflect the cortical functionality for neuronal oscillations. To investigate its potential for preclinical risk assessment, we evaluated the effects of the psychotogenic drugs, a non-competitive NMDA antagonist MK-801, a dopamine transporter inhibitor methylphenidate and a 5HT2 agonist DOI, on high gamma ASSR from rat auditory cortex using a chirp-modulated tone whose modulation frequency change from 1 to 120 Hz or from 120 to 1 Hz. The effect of MK-801 was very robust that it attenuated the ASSR (83 % reduction in power (<em>p</em> &lt; 0.0001) and 47 % reduction in phase-locking factor (PLF. <em>p</em> &lt; 0.001)) at 1 mg/kg dose. The effect of methylphenidate on ASSR in power was relatively mild while it significantly attenuated PLF by 36 % at 50 mg/kg. The effect of DOI was somewhat different: The administration of 3 mg/kg did not attenuate the ASSR amplitude, but instead increased the chirp-frequency independent high gamma band (power 322 % of control, <em>p</em> &lt; 0.05). These observations resemble the attenuation of phase-locked response and augmentation of non-phase-locked response in schizophrenia patients. In baseline period, the administration of 1 mg/kg of MK-801 or 50 mg/kg of methylphenidate enhanced spontaneous high gamma power (181 % of control for MK-801, p &lt; 0.001, 183 % of control for methylphenidate, p &lt; 0.05), which is also observed in human psychosis. Together with the clinical reports examining the drug effects on ASSR and with pharmacokinetic models from the literatures, our results suggest that ASSR and the related EEG response in high gamma band is a promising translational biomarker for the drug-induced psychosis, and that the use of chirp tone enables the clear discrimination of modulation frequency-dependent and -independent effects.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107839"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095377","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":"Comparison of multiple concentration-QTc modeling approaches","authors":"Jill Dalton ,&nbsp;Steve Denham ,&nbsp;Matthew St Peter ,&nbsp;Kevin Norton","doi":"10.1016/j.vascn.2025.107775","DOIUrl":"10.1016/j.vascn.2025.107775","url":null,"abstract":"<div><div>The ICH E14/S7B Q&amp;A guidance for <em>in vivo</em> preclinical studies recommends exposure-response modeling to support the assertion that there were no changes in QTc, to better characterize observed QTc prolongation, and/or when QTc changes are anticipated based on hERG assay results. While the guidance indicates that there are multiple acceptable approaches for concentration-QTc modeling, associated training documents and general industry views indicate that assessments should ideally occur in a separate phase of the study utilizing the same animals as those used for telemetry monitoring to generate a complete exposure profile. However, this “gold standard” approach is not necessarily ideal as it requires additional cost, time, and test material. Alternative options have been proposed including partial toxicokinetic profiles utilizing the same animals as those used for telemetry monitoring or by generating full profiles using data collected from a separate cohort of animals. However, it is unclear as to how concentration-QTc data estimates might differ with alternate approaches. The current study utilized moxifloxacin at 10, 30 and 90 mg/kg, PO in beagle dogs using a 4 × 4 cross-over design monitoring QTc via telemetry over 24 h. Plasma concentrations were determined at 0, 2, 4, 6, 8, 12, and 24 h postdose by using the same animals in a separate phase of the telemetry study or by using a separate cohort of animals. The 10 msec prolongation prediction values were estimated for the partial toxicokinetic profiles using data pairs to estimate plasma levels at each time point using the plasma samples collected during the telemetry phase. The actual exposure values were used for the independent groups. Additionally, simulations of possible toxicokinetic curves for the separate groups were used to estimate representative population values. The 10 msec prolongation prediction values varied minimally, with &lt;300 ng/ml difference among the various estimation methods. Therefore, comparison of these exposure-response modeling methods illustrates the relatively small degree of impact that the different approaches have on QTc prediction values as compared to the gold standard methodology, thus providing multiple viable study design options for researchers to consider in their preclinical testing strategies.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107775"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095376","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":"“All-inclusive” evaluation of the efficacy and safety of methotrexate in a murine breast cancer model integrating the 3Rs to enhance preclinical assessment","authors":"Tristan Rupp,&nbsp;Sonia Goineau,&nbsp;Guillaume Froget,&nbsp;Kendall Walker","doi":"10.1016/j.vascn.2025.107768","DOIUrl":"10.1016/j.vascn.2025.107768","url":null,"abstract":"<div><div>Safety pharmacology evaluation plays a crucial role in the preclinical assessment of anti-cancer drugs, ensuring their tolerability and minimizing potential adverse effects before clinical translation. While there is broad consensus around the importance of safety assessment in cancer drug evaluation at the clinical stage, this is poorly investigated at the preclinical level. This study aims to comprehensively evaluate the safety pharmacological properties of Methotrexate, a folate antagonist, in a preclinically relevant murine model of breast cancer and emphasizing the interest of such approach for 3Rs (Replacement, Reduction, and Refinement) in animal research. Female BALB/c mice were orthotopically implanted with 4 T1 mouse mammary carcinoma cells to establish breast cancer tumors. The mice were randomized into treatment or control groups. Methotrexate was injected at 25 and 1000 mg/kg (slow i.v. once a week for 3 weeks). Tumor growth kinetics, tumor volume, metastatic potential, hematological profile, and overall survival were assessed. Additionally, respiratory (whole body plethysmography) and behavioral (Irwin) functions were investigated longitudinally over four different timepoints to monitor the adverse effects associated with Methotrexate treatment. Interestingly, this approach aligns with the 3Rs by using an “all-inclusive” model that reduces the number of animals needed through the longitudinal assessment of multiple efficacy and safety parameters within the same study. This global approach minimizes potential risks prior to clinical development and provides valuable insights into the pharmacological properties of drugs for cancer therapy while adhering to ethical standards in animal research.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"135 ","pages":"Article 107768"},"PeriodicalIF":1.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145094211","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}