Journal of pharmacological and toxicological methods最新文献_第5页

The translation of QTc across species - Impact of subject number and exposure multiple tested on discriminatory sensitivity QTc的跨物种翻译——受试者数量和暴露倍数对区分敏感性的影响

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-01 DOI: 10.1016/j.vascn.2025.107774

Derek J. Leishman

Small, relatively insensitive studies can be useful in safety assessment when multiples of the therapeutic clinical concentration are tested. This is a fundamental principle in safety testing in animals which is equally valid for early clinical evaluations in healthy volunteers. It is often less practical to increase the number of test subjects than to increase the exposure tested. Both, when combined with the analysis method, can have an impact on the sensitivity to detect an effect. The objective is that the relationship between statistical power, analysis method, number of animals and exposure multiple explored can be illustrated using the example of QTc assessment in animals. The statistical power to detect an effect on the electrocardiogram QTc interval in nonhuman primates (NHP) for different analyses methods was known. The concentration-QTc relationship was also known for reference agents in NHP. Lastly, the critical concentration associated with a 10 ms QTc interval change in man was known for these same reference agents. This information was combined to illustrate how doubling the number of NHP used or increasing the exposure tested would support a conclusion concerning the presence or absence of an effect on the QTc interval for a test agent. In NHP, the most sensitive analysis methods have >80 % power (at p < 0.05) to detect an effect of the reference agent at the critical concentration using only 4 animals. Less sensitive techniques can detect an effect with the same power when either more animals are used or where higher multiples of the critical concentration are tested. This illustrates the principle that even with only 4 animals and an insensitive technique an effect can be detected provided higher exposures are tested. Conversely, a study using more animals, or a more sensitive analysis needn't require a higher exposure in animals to exclude an effect in man. Rather than focusing on a fixed QTc threshold sensitivity regardless of experimental design these analyses demonstrate that investigators have the flexibility to use the simplest available combination of exposures, animal numbers and analysis to achieve an effective QTc assessment.

小规模的、相对不敏感的研究可以用于安全性评估，当测试治疗性临床浓度的倍数时。这是动物安全试验的基本原则，同样适用于健康志愿者的早期临床评估。增加测试对象的数量往往比增加被测试的暴露程度更不实际。当两者与分析方法相结合时，会对检测效果的灵敏度产生影响。目的是通过动物QTc评估的例子来说明统计功效、分析方法、动物数量和所探讨的暴露倍数之间的关系。已知不同分析方法对非人灵长类动物（NHP）心电图QTc间期影响的统计能力。浓度- qtc关系在NHP的参考药物中也是已知的。最后，已知这些相同的参考药物与人类10 ms QTc间隔变化相关的临界浓度。将这些信息结合起来说明，将使用的NHP的数量加倍或增加所测试的暴露量将如何支持有关对测试剂的QTc间隔是否存在影响的结论。在NHP中，仅使用4只动物，最敏感的分析方法在检测临界浓度下对照剂的影响时具有>；80 %的功率（p <； 0.05）。当使用更多的动物或在测试临界浓度的更高倍数时，灵敏度较低的技术可以以相同的功率检测到效果。这说明了即使只有4只动物和一种不敏感的技术，只要测试更高的暴露量，也可以检测到效果的原理。相反，一项使用更多动物的研究，或者一项更敏感的分析，不需要在动物中使用更高的剂量来排除对人类的影响。这些分析表明，研究人员可以灵活地使用暴露、动物数量和分析的最简单的可用组合来实现有效的QTc评估，而不是将重点放在固定的QTc阈值灵敏度上。

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引用次数: 0

Comprehensive protocol for culturing and functionally characterizing primary mixed neural cells from the neonatal rat cortex 从新生大鼠皮层中培养和功能表征初级混合神经细胞的综合方案。

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-01 DOI: 10.1016/j.vascn.2025.108390

Thaynan Silva Ramos , Matheus Heidemann Tempone , Hercules Rezende Freitas , Monique Fonseca-Teixeira , Patricia Fernanda Schuck , Ricardo Augusto de Melo Reis , Gustavo Costa Ferreira

In vitro models using purified neurons or glial cells are crucial for studying neurological functions but often overlook intercellular interactions. Mixed neural cell cultures offer a more physiologically relevant system by preserving cell-to-cell communication and providing deeper insights into neural behavior. Here, we present a protocol for culturing mixed primary cells from the neonatal rat cerebral cortex and functionally characterizing them via calcium imaging. This method enables cell phenotyping, spatial distribution analysis, and activity monitoring in response to stimuli. Our model maintained a cellular composition resembling the native rat cortex, with 35.4 % neurons, 44.3 % astrocytes, and 20.3 % other cell types. Calcium imaging showed that ATP (100 μM) and BzATP (100 μM) evoked stronger calcium transients than KCl (50 mM). BzATP induced a sustained response mediated by P2X7 receptor activation, while ATP activated a broader range of P2 receptors. Unlike purified or enriched cultures, this mixed-cell system better replicates the cellular environment of the brain, ensuring reproducibility and biological relevance. This protocol provides a straightforward platform for investigating neuron-glia interactions and neural signaling, bridging the gap between simplified in vitro models and the complexity of neural networks. Its applications may advance research into neurobiological disease mechanisms and therapeutic development.

使用纯化的神经元或神经胶质细胞的体外模型对于研究神经功能至关重要，但往往忽略了细胞间的相互作用。混合神经细胞培养通过保持细胞间的通信和对神经行为的更深入的了解，提供了一个更生理相关的系统。在这里，我们提出了一种从新生大鼠大脑皮层培养混合原代细胞的方案，并通过钙成像对它们进行功能表征。这种方法使细胞表型，空间分布分析和活动监测响应刺激。我们的模型保持了与天然大鼠皮层相似的细胞组成，神经元占35.4% %，星形胶质细胞占44.3% %，其他细胞占20.3% %。钙成像显示ATP（100 μM）和BzATP（100 μM）比KCl（50 mM）诱发更强的钙瞬变。BzATP诱导了P2X7受体激活介导的持续反应，而ATP激活了更广泛的P2受体。与纯化或富集培养不同，这种混合细胞系统更好地复制了大脑的细胞环境，确保了可重复性和生物学相关性。该协议为研究神经元-胶质细胞相互作用和神经信号提供了一个简单的平台，弥合了简化的体外模型和神经网络复杂性之间的差距。它的应用可能会推动神经生物学疾病机制和治疗发展的研究。

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引用次数: 0

Elucidating the challenges with categorizing drugs that block cardiac sodium channels − The impact of temperature on drug-cardiac sodium channel interactions 阐明阻断心脏钠通道的药物分类的挑战-温度对药物-心脏钠通道相互作用的影响

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-01 DOI: 10.1016/j.vascn.2025.107805

Huimei Yu, Claudia Alvarez Baron, Jun Zhao, Jose Vicente, Lars Johannesen, Wendy W. Wu, Donglin Guo

Class I antiarrhythmic drugs (AADs) are categorized into Class IA, IB and IC subgroups based on their distinct electrophysiological consequences on the heart that are thought to arise from distinct interaction characteristics with cardiac sodium channels (Na_V). The original categorization centered on assessing electrophysiological features including drug effects on the maximal upstroke velocity of action potential (AP), AP duration, and effective refractory period using ventricular preparations from animals performed at physiological temperature (PT). Subsequently, categorization assessing drug binding and unbinding kinetics in patch clamp assays in Na_V overexpressing cells at room temperature (RT) was proposed. Temperature is known to impact Na_V gating, hence potentially affecting use- and state-dependence of drug interactions. Whether RT patch clamp data adequately capture drug-Na_V interaction characteristics that allows for the same subgroup categorization as in the original categorization using native tissues is unclear. Similarly, a systematic assessment of the effects on other cardiac ion channels following best practices is also lacking. This study characterized drug-Na_V (Na_V1.5) interaction characteristics of quinidine (IA), mexiletine (IB) and flecainide (IC) at near PT and RT using manual patch clamp. Additionally, drug's potencies on inhibiting the late Na_V1.5, hERG, and Ca_V1.2 currents were conducted at near PT following ICH S7B Q&A 2.1 best practices. At RT, use-dependent block of Na_V1.5 currents and unbinding kinetics were fastest for mexiletine, followed by quinidine, and flecainide. At near PT, use-dependent block and unbinding kinetics of quinidine remained faster than flecainide. Surprisingly, mexiletine showed no use-dependent block and no apparent unbinding at near PT. Preliminary assessment revealed that the three drugs have different effects on other cardiac ionic currents. The three Class I AADs showed distinct interaction characteristics with Na_V1.5 currents. Importantly, temperature-dependent binding and unbinding kinetics for mexiletine and different effects on other cardiac ionic currents for all three drugs were observed. These data provide mechanistic insights to drug-induced changes in myocyte APs that led to Class I AADs categorization, and lay a foundation for future research to identify the commonalities and differences of Class I AADs interacting with Na_V1.5 channels.

一类抗心律失常药物（AADs）被分为IA类、IB类和IC类，基于它们对心脏的不同电生理后果，这些后果被认为是由与心脏钠通道（NaV）的不同相互作用特征引起的。最初的分类集中于评估电生理特征，包括药物对动作电位（AP）最大上冲程速度的影响，AP持续时间，以及在生理温度（PT）下使用动物心室制剂的有效不应期。随后，在室温（RT）下，提出了膜片钳法在NaV过表达细胞中评估药物结合和解除结合动力学的分类。已知温度会影响导航门控，因此可能影响药物相互作用的使用和状态依赖性。RT膜片钳数据是否充分捕获药物- nav相互作用特征，从而允许与使用原生组织的原始分类相同的亚组分类尚不清楚。同样，对其他心脏离子通道影响的系统评估也缺乏最佳实践。本研究采用手动膜片钳对奎尼丁（IA）、美西汀（IB）和氟卡奈（IC）在PT和RT前后的药物- nav （NaV1.5）相互作用特征进行了研究。此外，药物抑制晚期NaV1.5、hERG和CaV1.2电流的效力在接近PT时按照ICH S7B Q&；A 2.1最佳实践进行。在RT下，使用依赖性的NaV1.5电流阻滞和解结合动力学最快的是美西汀，其次是奎尼丁和氟屈奈。在接近PT时，奎尼丁的使用依赖阻滞和解结合动力学仍然比氟喹奈快。令人惊讶的是，美西汀在PT附近没有表现出使用依赖性阻滞和明显的解结合。初步评估显示，这三种药物对其他心脏离子电流有不同的影响。3种I类AADs与NaV1.5电流表现出明显的相互作用特征。重要的是，观察了美西汀的温度依赖性结合和解结合动力学以及这三种药物对其他心脏离子电流的不同影响。这些数据为药物诱导的肌细胞APs变化导致I类AADs分类提供了机制见解，并为进一步研究识别与NaV1.5通道相互作用的I类AADs的共性和差异奠定了基础。

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引用次数: 0

Application for drug evaluation of human-induced pluripotent stem cell-derived cardiomyocytes cultured by anisotropic orientation 各向异性培养人诱导多能干细胞来源心肌细胞的药物评价应用

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-01 DOI: 10.1016/j.vascn.2025.107807

Hisako Tokuno , Hidenori Hiranuma , Akari Hasegawa , Ayano Satsuka

Human-induced pluripotent stem cell-derived cells (hiPSC) are expected to have applications in the fields of regenerative medicine and drug discovery. Due to the recent trend of reducing or eliminating animal testing, hiPSC is also expanded to use as the alternative method for animal testing. Human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) are especially effective as an alternative assay of human cardiomyocytes which are not easily available. hiPS-CMs are, however, considered immature, hence the maturation of hiPS-CMs is an important issue for proper assessment of drug response. To induce maturation of hiPS-CMs, various methods have been developed, including long-term culturing, three-dimensional (3D) tissue engineering, mechanical loading, electrical stimulation, modulation of substrate stiffness, and treatment with neurohormonal factors. In these context, we attempted an anisotropic orientation culture, in which the cultured cells are aligned in one orientation resembling in vivo cardiomyocytes. Culture substrate for anisotropic orientation culture was developed by applying our original nanofabrication technology (CellArray-Heart™, Oji Holdings Corporation), and the alignment and maturation of hiPS-CMs were evaluated with the culture substrate. The hiPS-CMs cultured in anisotropic orientation on CellArray-Heart™ showed elongated morphology and unidirectional contraction using cell motion vector analysis and increase of mRNA related to maturation of hiPS-CMs including KCNJ2, ATP2A2, and GJA1. Furthermore, in Ca²⁺ transient assay with isoproterenol (0.1 μM, 1 μM), the hiPS-CMs cultured in anisotropic orientation showed 1.5- to 2.4-fold change in beating rate and 1.1- to 1.4-fold change in amplitude, the parameters of contraction force, compared to hiPS-CMs treated with DMSO. The hiPS-CMs cultured in anisotropic orientation might have induced the expression of positive inotropy by beta-receptor stimulation. These results indicate the hiPS-CMs cultured in anisotropic orientation on CellArray-Heart™ are induced maturation and are suitable for drug evaluation.

人类诱导的多能干细胞衍生细胞（hiPSC）有望在再生医学和药物发现领域得到应用。由于近年来减少或消除动物试验的趋势，hiPSC也被扩展为动物试验的替代方法。人类诱导的多能干细胞衍生的心肌细胞（hiPS-CMs）作为一种不容易获得的人类心肌细胞的替代测定方法特别有效。然而，hiPS-CMs被认为是不成熟的，因此hiPS-CMs的成熟是正确评估药物反应的重要问题。为了诱导hiPS-CMs成熟，已经开发了多种方法，包括长期培养，三维（3D）组织工程，机械加载，电刺激，调节底物刚度以及神经激素因子处理。在这种情况下，我们尝试了各向异性取向培养，其中培养的细胞在一个类似于体内心肌细胞的方向上排列。各向异性取向培养的培养底物采用我们独创的纳米制造技术（CellArray-Heart™，Oji Holdings Corporation）开发，并利用培养底物评估hiPS-CMs的对准性和成熟度。细胞运动矢量分析显示，在CellArray-Heart™上各向异性培养的hiPS-CMs形态延长，单向收缩，与KCNJ2、ATP2A2和GJA1等hiPS-CMs成熟相关的mRNA增加。此外，在异丙肾上腺素（0.1 μM, 1 μM）的Ca2+瞬态分析中，与DMSO处理的hiPS-CMs相比，各向异性培养的hiPS-CMs的跳动速率变化了1.5 ~ 2.4倍，收缩力参数振幅变化了1.1 ~ 1.4倍。各向异性培养的hiPS-CMs可能通过β受体刺激诱导正性肌力的表达。这些结果表明，在CellArray-Heart™上各向异性培养的hiPS-CMs是诱导成熟的，适合用于药物评估。

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引用次数: 0

Development of drug toxicity evaluation platform using hiPSC-derived cardiac organoids hipsc源性心脏类器官药物毒性评价平台的建立

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-01 DOI: 10.1016/j.vascn.2025.107809

Hyun-lee Lee , Ji-hye Park , Hyun-su Kang , Ki-suk Kim

Cardiotoxicity is a critical aspect of safety evaluation in drug development. Traditional cardiotoxicity assessment methods rely primarily on animal models and 2D cell culture systems, which fail to replicate the complex physiological characteristics of human cardiac tissue fully. This study aims to generate human Cardiac Organoids (hCOs) derived from human induced Pluripotent Stem Cells (hiPSCs) and to use them to enhance the sensitivity of drug testing. hCOs were successfully cultured for up to 12 weeks, with a stable increase in heart rate observed over the cultivation period. Differentiation conditions were optimized by confirming the expression of cardiac markers (TNNT2), smooth muscle cell markers (aSMA), fibroblast markers (VIM), and endothelial markers (PECAM). The differentiation rate into cardiomyocytes was higher than that of conventional 2D cell culture methods. In calcium imaging using the positive drug nifedipine, the intensity of the calcium signal response of hCOs was confirmed to change depending on the concentration (1, 5, 10uM). This can be inferred that hCOs well reflects changes in various ion channels. Utilizing optimized hCOs conditions, we measured the changes in BPM induced by positive (Quinidine, Moxifloxacin, Nifedipine, E-4031), false positive (Diltiazem), false negative (Bepridil), and negative (Levofloxacin) drugs, comparing these to existing iCM data. The results demonstrated that hCOs exhibited more sensitive changes, suggesting that cardiac organoids can more sensitively and accurately reflect drug-induced cardiotoxicity than traditional 2D cell culture systems. This study presents a cardiotoxicity assessment platform using human-derived cardiac organoids. This approach can enhance the accuracy of cardiotoxicity assessment in the early stages of drug development, ultimately contributing to the development of safe and effective new drugs. Future research will measure electrophysiological changes to further optimize the evaluation platform and present an advanced cardiotoxicity evaluation platform.

This research was supported by a grant (22213MFDS391) from Ministry of Food and Drug Safety in 2024.

心脏毒性是药物开发中安全性评价的一个重要方面。传统的心脏毒性评估方法主要依赖于动物模型和二维细胞培养系统，无法完全复制人类心脏组织的复杂生理特征。本研究旨在利用人诱导多能干细胞（hiPSCs）制备人心脏类器官（hCOs），并利用其提高药物检测的敏感性。hco成功培养了12 周，在培养期间观察到心率稳定增加。通过确认心肌标记物（TNNT2）、平滑肌细胞标记物（aSMA）、成纤维细胞标记物（VIM）和内皮标记物（PECAM）的表达，优化分化条件。与传统的二维细胞培养方法相比，其向心肌细胞的分化率更高。在使用阳性药物硝苯地平的钙显像中，证实hCOs的钙信号响应强度随浓度（1,5,10 um）而变化。由此可以推断，hCOs很好地反映了各种离子通道的变化。利用优化的hCOs条件，我们测量了阳性（奎尼丁、莫西沙星、硝苯地平、E-4031）、假阳性（地尔硫卓）、假阴性（贝普利地尔）和阴性（左氧氟沙星）药物引起的BPM变化，并将其与现有的iCM数据进行比较。结果表明，hCOs表现出更敏感的变化，表明心脏类器官比传统的二维细胞培养系统更敏感、更准确地反映药物诱导的心脏毒性。本研究提出了一个使用人类来源的心脏类器官的心脏毒性评估平台。这种方法可以提高药物开发早期心脏毒性评估的准确性，最终有助于开发安全有效的新药。未来的研究将测量电生理变化，进一步优化评估平台，提出一个先进的心脏毒性评估平台。本研究于2024年获得国家食品药品安全部资助（22213MFDS391）。

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引用次数: 0

Pair housing large adult male cynomolgus monkeys is possible!, the use of telemetry to assess the benefits 成对住房的大型成年雄性食蟹猴是可能的！，利用遥测技术来评估效益

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-01 DOI: 10.1016/j.vascn.2025.107769

Peter Harris , Rosemary Santos , Thi Dong-Binh Tran , Dingzhou Li , Jan Bernal , Steven Kreuser , Erin Ricciardi , Siri Skowronek , Kiran Palyada , John Capitanio

Long term pair housing of sexually mature, cynomolgus monkeys in male-male pairs is a common practice based on social needs of the species and research design. This housing paradigm does include risks of aggression, trauma, and impact to studies. Some facilities may forgo attempting to socially house males due to these risks. We sought to leverage the often-easier task of male-female pairing to create longer term, stable pairs as an option to address the social housing needs of sexually mature males. To understand the risk/benefit of this practice, seven vasectomized male monkeys previously implanted with telemetry devices were housed with female counterparts. These animals were selected based on good health, and a history of single housing in a male only study room. The pairing process was gradual with increasing levels of interactions between the male and female occurring over 45 days. At various stages of the pairing process, telemetry data was collected to characterize heart rate, blood pressure, activity, and temperature with data averaged into 2 timebins, 12 am–4 am and 2 pm–6 pm. Initial telemetry was collected when female monkeys were not present in the room and was used for baseline values. A second and third study with different male monkeys were conducted to assess male-male pairing and the influence of cage size on the parameters collected previously. When the females were paired with the males, decreases in blood pressure (−9 mmHg), and increases in body temperature (+0.7 °C) were observed. No significant pairing effects were found for heart rate or physical activity. During the second study, when male animals were re-paired with their male counterparts, blood pressure decreased (−5 mmHg) similar to the male – female pairing study. In the third study, no significant cage size effect was noted on physiologic parameters. These reductions in blood pressure indicate a lower stress in paired animals unrelated to sex or cage size. This further supports the benefits of pair housing monkeys in a research environment. In conclusion, pairing vasectomized male monkeys with female monkeys is a viable option for long term telemetry colonies using single housed male animals.

性成熟的食蟹猴以雄性-雄性配对长期居住是基于物种的社会需求和研究设计的一种常见做法。这种住房模式确实包含了攻击、创伤和对研究的影响的风险。由于这些风险，一些机构可能会放弃为男性提供社交场所。我们试图利用通常更容易的男女配对来创造长期稳定的配对，作为解决性成熟男性社会住房需求的一种选择。为了了解这种做法的风险/收益，7只输精管切除的雄猴被植入了遥测装置，并与雌猴一起饲养。这些动物的选择是基于健康状况良好，并且有单独居住在男性自习室的历史。配对过程是渐进的，雄性和雌性之间的互动水平不断提高，持续了45 天。在配对过程的各个阶段，收集遥测数据来表征心率、血压、活动和温度，并将数据平均为2个时间区间，分别为12 am - 4 am和2 pm - 6 pm。最初的遥测数据是在雌性猴子不在房间时收集的，并用于基线值。在第二次和第三次研究中，我们用不同的雄猴来评估雄猴配对以及笼子尺寸对先前收集的参数的影响。当雌性与雄性配对时，观察到血压下降（- 9 mmHg），体温升高（+0.7 °C）。在心率和体力活动方面没有发现显著的配对效应。在第二项研究中，当雄性动物与雄性动物重新配对时，血压下降（- 5 mmHg），与雌雄配对研究相似。在第三项研究中，笼型尺寸对生理参数没有显著影响。这些血压的降低表明，与性别或笼子大小无关的成对动物的压力较低。这进一步支持了在研究环境中成对居住猴子的好处。综上所述，结扎后的雄猴与雌猴配对是一种可行的选择，用于长期遥测群体使用单房雄猴。

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引用次数: 0

QTc least significant difference in stand-alone implanted telemetry safety pharmacology and jacketed external telemetry regulatory toxicology studies in dogs QTc在独立植入式遥测安全药理学和外套遥测调节毒理学研究中差异最小

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-01 DOI: 10.1016/j.vascn.2025.107770

Abdel-Ilah El Amrani , Ana Gueorguieva-Apostolova , Camille Lagard , Anson Phillips , Francine El Amrani Callens

The use of least significant difference (LSD) as the metric for variability of specific study, especially in the context of ICH E14/S7B Q&As studies, is considered as part of best practice. This parameter represents not only the threshold of statistical significance for a given parameter, but also reflects the intrinsic study variability. LSD is commonly used to evaluate the variability of QTc changes in standalone 4 × 4 cross-over (XO) Q&As studies. To our knowledge, LSD evaluations are not commonly performed when jacketed external telemetry (JET) in parallel groups is used during safety pharmacology studies integrated in regulatory toxicology studies, despite the complementarity between this approach and standalone XO safety pharmacology studies. The use of one and/or the other approach could prove necessary depending on the nature and/or the PK/PD of the drug candidate. Therefore, we calculated LSDs for QT and QTc in 7 randomly selected telemetry investigations, including 3 XO design studies and 4 JET studies in a parallel design. Four male dogs and 32 dogs (16 males and 16 females) were used in each XO and JET investigation, respectively. Animals received vehicle with no impact on ECG parameters and/or 3 doses of test items. LSD is calculated using repeated measure ANOVA over 24 h in several intervals. LSDs for QT interval per study were 5 ± 2, 12 ± 5 and 8 ± 3 msec in XO and 11 ± 6, 8 ± 3, 13 ± 6 and 8 ± 4 msec in JET. LSDs for QTc (with Miyazaki correction) per study were 4 ± 2, 5 ± 2 and 5 ± 2 msec in XO and 3 ± 1, 6 ± 3, 12 ± 6 and 2 ± 1 msec in JET. Mathematical sensitivity comparisons between both approaches might be difficult. However, the threshold for detecting statistically significant increases in QT/QTc from JET cardiovascular studies was close to that shown in XO studies. This investigation shows that jacketed ECG can be as adequate as cross-over implanted cardiovascular studies to support an integrative risk assessment of QT prolongation in the dog. However, this first set of LSD values will need to be completed with the same analysis, based on a larger cohort of studies.

使用最小显著性差异（LSD）作为特定研究可变性的度量，特别是在ICH E14/S7B q&&； as研究的背景下，被认为是最佳实践的一部分。该参数不仅表示给定参数的统计显著性阈值，而且反映了研究的内在变异性。LSD通常用于评估独立4 × 4交叉（XO） Q&；As研究中QTc变化的可变性。据我们所知，尽管这种方法与独立的XO安全药理学研究具有互补性，但在纳入监管毒理学研究的安全药理学研究中，在平行组中使用夹带外部遥测（JET）时，通常不会进行LSD评估。根据候选药物的性质和/或PK/PD，可能需要使用一种和/或另一种方法。因此，我们在7个随机选择的遥测研究中计算了QT和QTc的lsd，包括3个XO设计研究和4个并行设计的JET研究。每次XO和JET调查分别使用4只公犬和32只犬（公16只、母16只）。动物接受对心电图参数无影响的载药和/或3个剂量的测试项目。LSD的计算使用重复测量方差分析超过24 h在几个间隔。每个研究lsd对QT间隔5 ± 2 12 ± 5  8 ± 3 msec XO和11 ± 6 8 ± 3,13 ± 6和8  ± 4 msec喷气机。(宫崎骏校正)/ lsd对高职院校学前教育专业研究4 ± 2 5 ±2  和 5 ± 2 msec XO和3 ± 1 6 ± 3、12 ± 6和2  ± 1 msec喷气机。两种方法之间的数学敏感性比较可能很困难。然而，JET心血管研究中QT/QTc统计学显著增加的阈值与XO研究中显示的阈值接近。这项研究表明，夹带心电图可以与交叉植入心血管研究一样充分，以支持犬QT间期延长的综合风险评估。然而，第一组LSD值将需要基于更大的研究队列进行相同的分析来完成。

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引用次数: 0

QTc data sensitivity: Is animal screening important to ensure reduced data variability? QTc数据敏感性：动物筛选对确保减少数据变异性重要吗？

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-01 DOI: 10.1016/j.vascn.2025.107787

Heather C. Heckt, Douglas E. Regalia, London E. Maberto, Stephen D. Tichenor

The ICH E14/S7B Q&As recommend a positive control study to demonstrate the assay sensitivity at a Test Site to detect changes in ECG parameters, namely QTc to enable translatability to the clinic. The objective of this study was to demonstrate the effectiveness of QT prolongation detection using telemetry implants (Physiotel: Data Sciences International) and manually restrained (snapshot) technology in non-human primates (NHP) dosed with moxifloxacin. Two Latin square positive control groups (cohorts 1 and 2) were simultaneously dosed (i.e., same room, drug formulation, technical staff, etc.) with moxifloxacin; 8 NHPs implanted with either the M-11 telemetry device or the L-21 telemetry device. Doses occurred every 7 days for a total of 4 doses and cardiovascular data were collected for 24 h on each occasion on unrestrained animals. Both groups showed measurable QT/QTc prolongation but a higher than anticipated least significant difference (LSD) of 22.4 msec for cohort 1 and 12.60 msec for cohort 2. It was determined that some of the animals had inherently longer QTc intervals than other animals leading to larger individual animal variability. Both cohorts had a maximum standard deviation (SD) of 25.68 and 14.62, respectively. To accurately determine sensitivity capabilities, subjects with similar QT intervals and the same dosing rotation were grouped together in a hybrid Latin square consisting of 2 animals from cohort 1 and 2 animals from cohort 2. The re-assigned animals with similar QT variability (maximum SD = 11.54) resulted in an LSD of 9.8 msec. Pre-screening subjects for inherent ECG waveform variability could allow for greater data sensitivity ensuring adherence to expected industry standard sensitivity. In addition to implanted telemetry, the same 8 subjects had manually restrained, snapshot data collected. All subjects demonstrated similar QT/QTc values from predose to postdose with no observation of QT variability (QTc of 281.4 to 272.7 msec respectively) and as such, could not detect measurable changes in QTc, compared to unrestrained techniques, which did provide measurable changes even with the highest calculated SD. Collectively, these data demonstrate diminished sensitivity of manually restrained, snapshot data as opposed to telemetry data from unrestrained, freely moving animals.

ICH E14/S7B q&&b推荐了一项阳性对照研究，以证明在测试现场检测ECG参数变化的检测灵敏度，即QTc，以使临床可翻译。本研究的目的是证明使用遥测植入物（Physiotel: Data Sciences International）和手动限制（快照）技术在服用莫西沙星的非人灵长类动物（NHP）中检测QT间期延长的有效性。两个拉丁方阳性对照组（队列1和2）同时给药（即同一房间、药物配方、技术人员等）莫西沙星；8只NHPs分别植入M-11遥测装置或L-21遥测装置。每7 天给药一次，共给药4次，在不受约束的动物身上，每次24 小时收集心血管数据。两组均显示可测量的QT/QTc延长，但高于预期的最小显著差异（LSD），队列1为22.4 msec，队列2为12.60 msec。结果表明，一些动物固有的QTc间隔比其他动物更长，导致动物个体差异更大。两个队列的最大标准差（SD）分别为25.68和14.62。为了准确地确定敏感性，将QT间期相似且给药轮换相同的受试者分组在一个混合拉丁方阵中，由来自队列1的2只动物和来自队列2的2只动物组成。重新分配的QT变异性相似的动物（最大SD = 11.54）导致LSD为9.8 msec。预先筛选受试者固有的ECG波形变异性可以允许更高的数据灵敏度，确保遵守预期的行业标准灵敏度。除了植入式遥测外，同样的8名受试者还被人工限制，快照数据收集。从给药前到给药后，所有受试者都表现出相似的QT/QTc值，没有观察到QT变异性（QTc分别为281.4至272.7 msec），因此，与不受约束的技术相比，无法检测到QTc的可测量变化，即使计算的SD最高，也能提供可测量的变化。总的来说，这些数据表明，与不受约束、自由移动的动物遥测数据相比，人工约束的快照数据的灵敏度降低了。

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引用次数: 0

Enhanced detection sensitivity in nonclinical cardiovascular telemetry studies: Impact of a characterized facility 增强检测灵敏度在非临床心血管遥测研究：一个特征设施的影响

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-01 DOI: 10.1016/j.vascn.2025.107789

Ty Speece, Zac Hawkins

In the context of a cardiovascular telemetry study, test facility characterization refers to the comprehensive assessment and description of the laboratory environment, equipment, procedures, and other relevant factors that could influence the outcomes and interpretation of the study. The goal is to ensure the integrity, reproducibility, and validity of telemetry data collected from implanted animals. Model verification, particularly regarding sensitivity analysis, is fundamental for ensuring the validity of these studies. In this investigation, we assessed the influence of transitioning to a purpose-built telemetry facility on model sensitivity, focusing on dofetilide-induced QT prolongation. Beagle dogs previously implanted with Data Sciences International ™ implants were relocated into a new facility, purpose built for telemetry studies. Following ICH E14/S7B guidelines, animals were administered dofetilide to induce QT prolongation. Sensitivity analysis, including the minimum detectable difference (MDD) and least significant difference (LSD) for the corrected QT interval (QTcI), was performed at both the previous facility and the newly established purpose-built facility. During baseline data collection, persistent signal dropout was observed in all animals resulting in data loss of ~20 % of total data collected. Troubleshooting steps included internal hardware and software diagnostics, battery assessment, and environmental assessments. The Received Signal Strength Indicator (RSSI) feature within the telemetry software was leveraged to pinpoint the source of interference. Data analysis revealed a consistent source of interference with an average strength of 30 (RSSI relative strength), coinciding with the operation of the facility's wireless security system (900 MHz), the same frequency band used for transmission and reception of telemetry data. Replacement of the security system with hardware operating at 433 MHz resulted in a negligible interference signal, with RSSI values averaging below 8, similar to previous facility values. An improvement in sensitivity at the purpose-built facility was observed. The MDD for QTcI prolongation was reduced to 6.2 ms, surpassing findings at previous facility (10.8 ms). Additionally, the LSD demonstrated enhanced sensitivity at the new facility (QTcI, 4.5 ms). Transitioning to a purpose-built telemetry facility has demonstrated tangible benefits in enhancing sensitivity. The optimized and characterized environment minimizes extraneous variables, enhancing the detection of QTcI prolongation induced by dofetilide.

在心血管遥测研究的背景下，测试设施表征是指对实验室环境、设备、程序和其他可能影响研究结果和解释的相关因素的综合评估和描述。目的是确保从植入动物身上收集的遥测数据的完整性、可重复性和有效性。模型验证，特别是敏感性分析，是确保这些研究有效性的基础。在这项研究中，我们评估了过渡到专用遥测设备对模型敏感性的影响，重点是多非他特引起的QT延长。之前植入Data Sciences International™植入物的Beagle犬被重新安置到一个新的设施中，专门用于遥测研究。根据ICH E14/S7B指南，给动物注射多非利特以诱导QT间期延长。敏感性分析，包括校正QT间期（QTcI）的最小可检测差异（MDD）和最小显著差异（LSD），在以前的设施和新建立的专用设施中进行。在基线数据收集期间，所有动物都观察到持续的信号丢失，导致数据丢失约20% %。故障排除步骤包括内部硬件和软件诊断、电池评估和环境评估。遥测软件中的接收信号强度指示器（RSSI）功能被用来精确定位干扰源。数据分析显示，干扰源平均强度为30 （RSSI相对强度），与设施无线安全系统（900 MHz）的运行相一致，该频段用于传输和接收遥测数据。将安全系统更换为运行频率为433 MHz的硬件，导致干扰信号可以忽略不计，RSSI值平均低于8，与以前的设施值相似。观察到专门建造的设施的灵敏度有所提高。QTcI延长的MDD降至6.2 ms，超过了之前设施的发现（10.8 ms）。此外，LSD在新设施中显示出更高的灵敏度（QTcI, 4.5 ms）。过渡到专门建造的遥测设施在提高灵敏度方面已经证明了切实的好处。优化和表征的环境最大限度地减少了外来变量，增强了对多非利特引起的QTcI延长的检测。

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引用次数: 0

Characterization of vasoactive agents in a canine safety pharmacology PKPD model: A HESI initiative 血管活性药物在犬安全药理学PKPD模型中的表征：HESI倡议

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-01 DOI: 10.1016/j.vascn.2025.107790

Yevgeniya E. Koshman , C. Michael Foley , Jennifer Pierson , Siddhartha Bhatt , Todd Wisialowski , Hugo A. Vargas , Peter Hoffmann , Kevin Norton , Eric I. Rossman , Mark A. Osinski , Kelly Ashcroft-Hawley , Michael K. Pugsley

Increases in arterial blood pressure (BP) contribute to adverse cardiovascular (CV) outcomes in patients; preclinical effects of a drug on BP are routinely evaluated during the safety pharmacology assessments as outlined in the ICH S7A guidance. A Health and Environmental Sciences Institute (HESI) Consortium initiated a multi-site study with the objective to assess the ability of the standard conscious telemetry instrumented CV dog model to detect drug-induced changes in BP and evaluate translation to human data. The goal of these studies is also to determine the reproducibility and consistency of BP assessment when measured across different laboratories using the same study protocol and recording methodology to detect drug-induced changes in hemodynamics using drugs known to clinically elevate and reduce BP. Animals were chronically instrumented with a BP catheter and ECG electrodes for telemetric collection of hemodynamic and ECG endpoints, respectively. Study endpoints include systolic, diastolic, and mean BP, heart rate, electrocardiogram (ECG), body temperature, and locomotor activity. Drugs evaluated include midodrine (alpha-1 agonist), nifedipine (calcium channel blocker), hydralazine (direct-acting smooth muscle relaxant), prazosin (alpha-1 blocker) and milrinone (phosphodiesterase-3 inhibitor). Drugs were selected based on known pharmacological mechanisms of action, primary cardiovascular effects as well as availability of clinical effect and exposure data. Drugs were evaluated in beagle dogs using a double (8 × 4) Latin square design and administered orally at 3 doses selected to match clinical exposure data with a vehicle control. A full pharmacokinetic profile for each drug was conducted in dogs at doses selected using automated blood sampling (ABS). Initial analysis shows that all 5 positive control drugs show consistent hemodynamic profiles (e.g., BP elevation or reduction) in the dog as seen in humans. These data sets with additional testing at multiple sites will be amenable to further statistical analysis, super-interval analysis and follow-up study endpoint evaluation such as pressure waveform analysis. The results from this chronically instrumented conscious dog model will provide essential information about accuracy and consistency in blood pressure measurement across multiple sites and translation of preclinical BP data to clinical outcomes.

动脉血压（BP）升高有助于患者不良心血管（CV）结局；在ICH S7A指南中概述的安全性药理学评估期间，对药物对BP的临床前作用进行常规评估。健康与环境科学研究所（HESI）联盟发起了一项多地点研究，目的是评估标准有意识遥测仪器CV狗模型检测药物引起的血压变化的能力，并评估其对人类数据的转化。这些研究的目的还在于确定在不同实验室使用相同的研究方案和记录方法测量血压评估的可重复性和一致性，以检测药物引起的血流动力学变化，使用已知的临床升高和降低血压的药物。动物长期使用血压导管和ECG电极，分别遥测收集血流动力学和ECG终点。研究终点包括收缩压、舒张压和平均血压、心率、心电图（ECG）、体温和运动活动。评估的药物包括米多宁（α -1受体激动剂）、硝苯地平（钙通道阻滞剂）、肼嗪（直接作用平滑肌松弛剂）、吡唑嗪（α -1受体阻滞剂）和米立酮（磷酸二酯酶-3抑制剂）。药物的选择是基于已知的药理作用机制、主要的心血管效应以及临床效果和暴露数据。采用双（8 × 4）拉丁方设计对比格犬进行药物评估，并选择三种剂量口服给药，以匹配临床暴露数据与载体对照。使用自动血液采样（ABS）在狗体内以选定剂量对每种药物进行了完整的药代动力学分析。初步分析显示，所有5种阳性对照药物在狗身上表现出与人类相同的血流动力学特征（例如血压升高或降低）。 这些数据集在多个地点进行额外的测试，将适用于进一步的统计分析，超区间分析和后续研究终点评估，如压力波形分析。 这个长期仪器化的有意识狗模型的结果将提供关于跨多个部位血压测量的准确性和一致性以及将临床前血压数据转化为临床结果的重要信息。

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