Journal of pharmacological and toxicological methods最新文献

Corrigendum to ‘PolyCheck: A hybrid model for predicting polypharmacy-induced adverse drug reactions in tuberculosis treatment using heterogenous drug-target-ADR networks’ [Journal of pharmacological and toxicological methods 136 (2025) 108393] “PolyCheck：一种混合模型，用于预测结核治疗中使用异质药物-靶点-不良反应网络的多种药物引起的药物不良反应”的更正[Journal of pharmacyand toxicological methods 136(2025) 108393]。

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-12-01 DOI: 10.1016/j.vascn.2025.108402

Ahmad Tamim Ghafari , Yuslina Zakaria , Mizaton Hazizul Hasan , Abu Bakar Abdul Majeed , Qand Agha Nazari

引用次数: 0

Non-clinical cardiovascular safety testing: Current status, gaps and emerging trends 非临床心血管安全检测：现状、差距和新趋势

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-10-22 DOI: 10.1016/j.vascn.2025.108403

Peter Hoffmann , Michael K. Pugsley

Cardiovascular adverse drug reactions remain a leading cause of drug attrition. They may emerge during non-clinical or clinical development and often remain undetected until post-marketing, prompting increased regulatory focus. Historically, non-clinical cardiovascular safety testing has centered on assessing QT interval prolongation and torsades de pointes risk, primarily via inhibition of the delayed rectifier potassium current IKr. Such focus may overlook broader cardiovascular liabilities affecting other parameters of the cardiovascular system.

This review explores the status and limitations of current non-clinical cardiovascular safety assessments, in particular the over-reliance on the core battery studies defined in ICH S7A and S7B and the insufficient use of mechanistic follow-up assessments. We highlight the gaps between the results of non-clinical cardiovascular safety testing and the emergence of cardiovascular adverse drug reactions in later clinical phases or real-world use. We conclude that to narrow the gaps, there is a need to advance non-clinical methods to detect and adequately measure adverse effects on cardiac rhythm, myocardial contraction, blood pressure, and thrombogenicity. The review further discusses emerging trends and challenges for improving translational relevance, including advanced in vitro and in vivo models, and proposes a re-evaluation of outdated regulatory frameworks to better address diverse cardiovascular risks. Emphasis is placed on functional and mechanistic endpoints over structural pathology, aligning non-clinical safety methodologies with clinical outcomes.

心血管不良反应仍然是药物损耗的主要原因。它们可能在非临床或临床开发期间出现，并且通常直到上市后才被发现，这促使监管部门加强了关注。从历史上看，非临床心血管安全性测试主要是通过抑制延迟整流钾电流IKr来评估QT间期延长和针尖扭转的风险。这种关注可能会忽略影响心血管系统其他参数的更广泛的心血管责任。本综述探讨了目前非临床心血管安全性评估的现状和局限性，特别是对ICH S7A和S7B中定义的核心电池研究的过度依赖以及机械随访评估的不足。我们强调了非临床心血管安全性测试结果与临床后期或实际使用中出现的心血管药物不良反应之间的差距。我们的结论是，为了缩小差距，有必要推进非临床方法来检测和充分测量对心律、心肌收缩、血压和血栓形成的不利影响。这篇综述进一步讨论了改善转译相关性的新趋势和挑战，包括先进的体外和体内模型，并提出了对过时的监管框架的重新评估，以更好地应对各种心血管风险。重点放在功能性和机械性终点，而不是结构病理学，将非临床安全性方法与临床结果结合起来。

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引用次数: 0

A novel, flexible, and accessible method for the ex vivo induction and quantification of excitotoxicity 一种新颖的、灵活的、可接近的体外诱导和定量兴奋毒性的方法。

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-10-06 DOI: 10.1016/j.vascn.2025.108401

Samantha J. Carew , Christiana M. Kennedy , Meghan L. Greenland, Matthew P. Parsons

Excitotoxicity is a key driver of neuronal death in diverse brain conditions, yet most toxicity assays rely on in vitro models that remove cells from their complex native environment within the brain parenchyma. Here, we present a novel ex vivo method to quantify N-methyl-d-aspartate (NMDA)-induced excitotoxicity using acute brain slices from male and female young adult c57bl/6 and aged B6129SF2J mice, similar to those used for conventional electrophysiological recordings. Acute hippocampal slices were recovered in an N-methyl-D-glucamine (NMDG)-based recovery solution, then treated with low-magnesium artificial cerebrospinal fluid (aCSF) containing the co-agonist glycine to promote receptor activation, with or without exogenous NMDA. Following treatment, slices were fixed, cryoprotected, and cryosectioned to 20 μm for immunohistochemistry. Apoptotic cell death was assessed by staining for cleaved caspase-3, and was combined with the percentage of dead space to calculate a toxicity index for overall excitotoxic cell death. Importantly, exposure to low-magnesium aCSF with glycine alone was sufficient to elevate active caspase-3 levels, an effect that was further enhanced by exogenous NMDA application and prevented by NMDAR antagonism. Our ex vivo method largely preserves the cytoarchitecture and local microenvironment of brain tissue, enabling the assessment of cell-specific vulnerabilities to excitotoxic damage in select brain regions at defined ages. It is particularly well-suited for use in neurodegenerative disease models, where excitotoxic susceptibility may evolve over time. In all, the approach described here provides a reliable and accessible alternative to dissociated cell cultures, bridging the gap between in vitro and in vivo systems for studying glutamate-induced cell death.

兴奋性毒性是多种脑条件下神经元死亡的关键驱动因素，然而大多数毒性试验依赖于体外模型，将细胞从脑实质内复杂的天然环境中移除。在这里，我们提出了一种新的离体方法来量化n -甲基-d-天冬氨酸（NMDA）诱导的兴奋毒性，使用雄性和雌性年轻成年c57bl/6和老年B6129SF2J小鼠的急性脑切片，类似于传统的电生理记录。急性海马切片在n -甲基- d -葡萄糖胺（NMDG）为基础的恢复液中恢复，然后用含有协同激动剂甘氨酸的低镁aCSF处理，以促进受体激活，有或没有外源性NMDA。处理后，固定切片，冷冻保护，冷冻切片至20 μm进行免疫组织化学。通过裂解caspase-3染色评估凋亡细胞死亡，并结合死亡空间百分比计算总体兴奋性毒性细胞死亡的毒性指数。重要的是，仅用甘氨酸暴露于低镁aCSF足以提高活性caspase-3水平，外源NMDA应用进一步增强了这一作用，并被NMDAR拮抗阻止。我们的离体方法在很大程度上保留了脑组织的细胞结构和局部微环境，从而能够评估特定年龄的特定大脑区域对兴奋毒性损伤的细胞特异性脆弱性。它特别适合用于神经退行性疾病模型，其中兴奋毒性易感性可能随着时间的推移而演变。总之，本文所描述的方法提供了一种可靠且可接近的解离细胞培养替代方法，弥合了体外和体内系统之间的差距，用于研究谷氨酸诱导的细胞死亡。

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引用次数: 0

Optimization strategy for modeling sick sinus syndrome in rats: Balancing effect and animal care 大鼠病窦综合征模型的优化策略：平衡效应与动物护理

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-25 DOI: 10.1016/j.vascn.2025.108399

Liming He , Xinxin Shi , Yucheng Wang , Shuwei Huang

Objective

This study aimed to validate the feasibility of establishing a sick sinus syndrome (SSS) rat model by injecting 3 % sodium hydroxide (NaOH) into the jugular vein at a rate of 0.01 mL/s, and to assess reductions in animal mortality and vascular injury.

Results

Compared with the conventional 10 % NaOH method, the modified 3 % NaOH approach yielded a significantly higher modeling success rate (78 % vs. 44 %), substantially lower mortality (15 % vs. 48 %), and reduced jugular vein injury. Electrophysiological evaluations demonstrated a time-dependent decrease in post-modeling heart rate (HR), characterized by widened P-waves and prolonged PR intervals. Additionally, heart rate variability (HRV) analysis revealed a notable increase in the standard deviation of normal-to-normal intervals (SDNN) 2 weeks post-modeling. Histopathological analysis (HE staining) indicated more pronounced necrosis and fibrosis in sinus node P-cells, accompanied by elevated levels of hydroxyproline (HYP) and transforming growth factor β1 (TGF-β1). Furthermore, assessments of pacing-related ion channels showed downregulated transcription and expression of hyperpolarization-activated cyclic nucleotide-gated channel 4 (HCN4) and reduced SCN5A transcription, aligning with observed electrophysiological abnormalities.

Conclusion

The modified 3 % NaOH method effectively establishes an SSS rat model, offering advantages in simplicity, cost-efficiency, and animal welfare. This model provides a practical platform for fundamental SSS research, especially in resource-limited laboratories.

目的探讨3%氢氧化钠（NaOH）以0.01 mL/s的速度注入颈静脉建立病窦综合征（SSS）大鼠模型的可行性，并观察其对动物死亡率和血管损伤的影响。结果与传统的10% NaOH方法相比，改良的3% NaOH方法的建模成功率显著提高（78%对44%），死亡率显著降低（15%对48%），并减少了颈静脉损伤。电生理评估显示建模后心率（HR）的时间依赖性下降，其特征是p波变宽和PR间隔延长。此外，心率变异性（HRV）分析显示，在建模后2周，正常到正常间隔（SDNN）的标准偏差显著增加。组织病理学分析（HE染色）显示窦结p细胞坏死和纤维化更为明显，并伴有羟脯氨酸（HYP）和转化生长因子β1 （TGF-β1）水平升高。此外，对起搏相关离子通道的评估显示，超极化激活的环核苷酸门控通道4 （HCN4）的转录和表达下调，SCN5A转录减少，这与观察到的电生理异常一致。结论改良的3%氢氧化钠法能有效地建立SSS大鼠模型，具有简便、经济、动物福利等优点。该模型为基础SSS研究提供了一个实用的平台，特别是在资源有限的实验室中。

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引用次数: 0

LC–MS/MS-based simultaneous quantification of chlorthalidone and cilnidipine in rat plasma: Pharmacokinetic evaluation, green analytical assessment, and DoE-driven optimization LC-MS/ ms同时定量大鼠血浆中氯噻酮和西尼地平：药代动力学评价、绿色分析评价和doe驱动优化。

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-25 DOI: 10.1016/j.vascn.2025.108400

Sravanthi Gandu, Kumaraswamy Gandla, Lalitha Repudi

A highly sensitive and reproducible liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was systematically developed and optimized using a Quality by Design (QbD) framework for the simultaneous quantification of chlorthalidone and cilnidipine in rat plasma. Critical method variables—organic phase composition, flow rate, and mobile phase pH—were identified through risk assessment and subsequently optimized via Box–Behnken Design to ensure analytical robustness. Optimal chromatographic conditions comprised 20 % organic content, a flow rate of 1.0 mL/min, and a mobile phase pH of 3.0, facilitating efficient resolution of both analytes. Detection was achieved in positive electrospray ionization mode using multiple reaction monitoring, with transitions of m/z 339.8909 → 85.0951 for chlorthalidone, m/z 493.5237 → 300.1587 for cilnidipine, and m/z 515.6423 → 342.6158 for telmisartan, employed as the internal standard. Method validation, performed in accordance with European Medicines Agency (EMA) guidelines, demonstrated excellent linearity (r² > 0.998), accuracy, and precision, with coefficient of variation consistently <10 %. The method exhibited strong analyte stability and was successfully applied to the pharmacokinetic evaluation of both drugs in Wistar rats. This DoE-optimized LC–MS/MS platform offers a selective, reliable, and environmentally conscious analytical solution for the preclinical assessment of chlorthalidone and cilnidipine.

建立了高灵敏度、高重复性的液相色谱-串联质谱（LC-MS/MS）同时定量测定大鼠血浆中氯噻酮和西尼地平的方法，并采用质量设计（QbD）框架进行了系统优化。通过风险评估确定了关键的方法变量——有机相组成、流速和流动相ph，随后通过Box-Behnken Design进行优化，以确保分析的稳健性。最佳色谱条件为有机含量为20% %，流速为1.0 mL/min，流动相pH为3.0，有利于两种分析物的高效分离。采用多反应监测的正电喷雾电离方式进行检测，以m/z 339.8909 → 85.0951为氯噻酮，m/z 493.5237 → 300.1587为西尼地平，m/z 515.6423 → 342.6158为替米沙坦内标。方法验证，按照欧洲药品管理局（EMA）指南进行，证明了良好的线性（r2 > 0.998），准确度和精密度，变异系数一致

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引用次数: 0

Exploratory safety investigations in normal, freely moving Göttingen Minipigs using telemetry: Pharmacological validation 使用遥测技术对正常、自由移动Göttingen迷你猪进行探索性安全性调查：药理学验证。

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-09 DOI: 10.1016/j.vascn.2025.108395

Julie Jacobsen, Berit Ø. Christoffersen

Introduction

Adverse cardiovascular (CV) effects is a major cause of drug attrition. Early assessment of CV risk for new drug candidates may be warranted for early de-risking of the further development. Predictive animal models and a careful study design are needed for decision-making. The aim of this study was to characterise the CV effects of three cardiometabolic compounds with known CV effects in humans – the GLP-1 receptor agonist liraglutide, the melanocortin receptor 4 agonist (MC4-RA) LY2112688 and urocortin-2 (UCN2) - in Göttingen Minipigs to evaluate the predictability of this model.

Materials and methods

Female Göttingen Minipigs with telemetry implants (n = 6–8) were used in 3 consecutive cross-over studies looking at CV effects of liraglutide, LY2112688 and UCN2. Main endpoints were: Mean arterial blood pressure (MAP), systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR), in addition to activity and body temperature.

Results

Liraglutide at the highest dose level of 3 nmol/kg (Day 7) induced a significant increase in 24 h HR (p < 0.01) compared to vehicle. No significant differences in MAP, SBP or DBP were observed. The MC4-RA LY2112688 at a dose level of 0.1–0.15 mg/kg (Day 4) gave rise to significant increases in all of 24 h HR (p < 0.05), MAP (p < 0.01), SBP (p < 0.01) and DBP (p < 0.05) compared to vehicle. UCN2 infusion resulted in a significant increase in HR (p < 0.05) and a significant decrease in SBP (p < 0.05).

Conclusion

The study highlights different CV study designs in Göttingen Minipigs and show that this model qualitatively reproduced the CV effects observed in humans following treatment with the three test compounds. These data support the minipig as a translational preclinical model for exploratory safety evaluations, although the magnitude of the changes may not translate completely between species.

不良心血管（CV）效应是药物损耗的主要原因。早期评估新的候选药物的心血管风险可能是必要的，以早期降低进一步开发的风险。决策需要预测性动物模型和仔细的研究设计。本研究的目的是在Göttingen迷你猪中表征三种已知心血管效应的心脏代谢化合物——GLP-1受体激动剂利拉鲁肽、黑素皮质素受体4激动剂（MC4-RA） LY2112688和尿皮质素-2 （UCN2）的心血管效应，以评估该模型的可预测性。材料和方法：使用携带遥测植入物的雌性Göttingen迷你猪（n = 6-8）进行连续3项交叉研究，观察利拉鲁肽、LY2112688和UCN2的CV效应。主要终点是：平均动脉压（MAP）、收缩压（SBP）、舒张压（DBP）和心率（HR），以及活动和体温。结果：利拉鲁肽最高剂量3 nmol/kg（第7天）诱导24 h HR显著增加（p ）。结论：该研究突出了Göttingen迷你猪的不同CV研究设计，并表明该模型在三种试验化合物治疗后对人类观察到的CV效应进行了定量再现。这些数据支持迷你猪作为探索性安全性评估的转化临床前模型，尽管变化的幅度可能无法完全在物种之间转换。

{"title":"Exploratory safety investigations in normal, freely moving Göttingen Minipigs using telemetry: Pharmacological validation","authors":"Julie Jacobsen, Berit Ø. Christoffersen","doi":"10.1016/j.vascn.2025.108395","DOIUrl":"10.1016/j.vascn.2025.108395","url":null,"abstract":"<div><h3>Introduction</h3><div>Adverse cardiovascular (CV) effects is a major cause of drug attrition. Early assessment of CV risk for new drug candidates may be warranted for early de-risking of the further development. Predictive animal models and a careful study design are needed for decision-making. The aim of this study was to characterise the CV effects of three cardiometabolic compounds with known CV effects in humans – the GLP-1 receptor agonist liraglutide, the melanocortin receptor 4 agonist (MC4-RA) LY2112688 and urocortin-2 (UCN2) - in Göttingen Minipigs to evaluate the predictability of this model.</div></div><div><h3>Materials and methods</h3><div>Female Göttingen Minipigs with telemetry implants (<em>n</em> = 6–8) were used in 3 consecutive cross-over studies looking at CV effects of liraglutide, LY2112688 and UCN2. Main endpoints were: Mean arterial blood pressure (MAP), systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR), in addition to activity and body temperature.</div></div><div><h3>Results</h3><div>Liraglutide at the highest dose level of 3 nmol/kg (Day 7) induced a significant increase in 24 h HR (<em>p</em> < 0.01) compared to vehicle. No significant differences in MAP, SBP or DBP were observed. The MC4-RA LY2112688 at a dose level of 0.1–0.15 mg/kg (Day 4) gave rise to significant increases in all of 24 h HR (<em>p</em> < 0.05), MAP (<em>p</em> < 0.01), SBP (p < 0.01) and DBP (p < 0.05) compared to vehicle. UCN2 infusion resulted in a significant increase in HR (<em>p</em> < 0.05) and a significant decrease in SBP (p < 0.05).</div></div><div><h3>Conclusion</h3><div>The study highlights different CV study designs in Göttingen Minipigs and show that this model qualitatively reproduced the CV effects observed in humans following treatment with the three test compounds. These data support the minipig as a translational preclinical model for exploratory safety evaluations, although the magnitude of the changes may not translate completely between species.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"136 ","pages":"Article 108395"},"PeriodicalIF":1.8,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145042691","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}

引用次数: 0

Introducing the Sinclair Nanopig™ model: Preliminary genomic, proteomic, and hepatic CYP450 characterization for (bio)pharmaceutical safety assessment 介绍Sinclair Nanopig™模型：用于（生物）药物安全性评估的初步基因组学，蛋白质组学和肝脏CYP450表征

IF 1.8 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods

Pub Date : 2025-09-06 DOI: 10.1016/j.vascn.2025.108394

Yafei Chen , Nathan Bivens , Hong An , Brian Mooney , Thao Nguyen , Lyndon Coghill , Jennifer Horkman , Lois Haupt , Melissa Evans , Rebecca Campbell , Wendell Davis

The Nanopig™ model is an emerging non-rodent platform for (bio)pharmaceutical safety assessment, with potential advantages for translational research. Here, we report initial characterization results using whole genome sequencing (WGS) and tissue-based proteomics, focusing on drug metabolism and immune system relevance. WGS produced a high-quality Nanopig™ genome assembly (2.8–2.9 Gb), with >98 % alignment to the Duroc pig reference genome, and identified key metabolic and immune-related genes, including 47 cytochrome P450 (CYP450) genes with high homology to human CYP450 families. Proteomic profiling of 15 pharmaceutically relevant tissues revealed human orthologous drug metabolism enzymes and transporters (DMETs), as well as immune-related proteins, indicating similarities to human CYP450 enzyme abundance and tissue distribution. Functional evaluation of hepatic CYP450 activity yielded kinetic parameters (K_m, V_max) in the range observed in humans and beagle dogs. These early findings represent a foundational multi-omics dataset for the Nanopig™, suggesting its future use as a translational model in preclinical safety assessment. This work provides an early framework for species selection strategies and model optimization, with the long-term goal of reducing reliance on traditional non-rodent species in drug development.

Nanopig™模型是一种新兴的（生物）药物安全性评估的非啮齿动物平台，具有转化研究的潜在优势。在这里，我们报告了使用全基因组测序（WGS）和基于组织的蛋白质组学的初步表征结果，重点关注药物代谢和免疫系统相关性。WGS生产了高质量的Nanopig™基因组组装（2.8-2.9 Gb），与杜洛克猪参考基因组有98%的一致性，并鉴定了关键的代谢和免疫相关基因，包括47个与人类CYP450家族高度同源的细胞色素P450 （CYP450）基因。15个药学相关组织的蛋白质组学分析揭示了人类同源药物代谢酶和转运蛋白（DMETs）以及免疫相关蛋白，表明与人类CYP450酶丰度和组织分布相似。肝脏CYP450活性的功能评估得到的动力学参数（Km, Vmax）在人类和比格犬观察到的范围内。这些早期发现代表了Nanopig™的基础多组学数据集，表明其未来可作为临床前安全性评估的转化模型。这项工作为物种选择策略和模型优化提供了一个早期框架，其长期目标是在药物开发中减少对传统非啮齿动物物种的依赖。

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引用次数: 0

PolyCheck: A hybrid model for predicting polypharmacy-induced adverse drug reactions in tuberculosis treatment using heterogenous drug-target-ADR networks PolyCheck：一个混合模型，用于预测结核治疗中使用异质药物-靶点-不良反应网络的多种药物引起的药物不良反应。

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.2025.108393

Ahmad Tamim Ghafari , Yuslina Zakaria , Mizaton Hazizul Hasan , Abu Bakar Abdul Majeed , Qand Agha Nazari

Polypharmacy during tuberculosis (TB) treatment, particularly in patients with comorbidities such as diabetes mellitus (DM), significantly increases the risk of adverse drug reactions (ADRs) due to complex drug–drug interactions (DDIs). Existing computational methods primarily focus on pairwise drug interactions, often failing to capture the multifactorial nature of ADRs in polypharmacy contexts. To address this gap, we developed PolyCheck, a hybrid predictive model that integrates network-based and rule-based methods to identify potential ADRs arising from multi-drug regimens. We constructed a heterogeneous Drug–Target–ADR interaction network comprising first-line anti-TB and antidiabetic drugs, their targets, and associated ADRs. The Random Walk with Restart (RWR) algorithm was employed to rank ADR nodes, and a rule-based layer further refined predictions by incorporating the biological relevance of Drug–Target–ADR associations. Evaluation using cross-validation and case-based testing demonstrated strong predictive performance, with accuracy, precision, recall, F1-score, and AUPRC values of 0.70, 0.74, 0.92, 0.81, and 0.74, respectively. PolyCheck offers a scalable and interpretable approach for predicting ADRs in complex treatment regimens and can support safer, individualized TB therapy in patients with comorbid conditions.

在结核病（TB）治疗期间，特别是在患有糖尿病（DM）等合并症的患者中，由于复杂的药物相互作用（ddi），多种药物治疗显著增加了药物不良反应（adr）的风险。现有的计算方法主要集中在药物的成对相互作用上，往往无法捕捉到多药环境下adr的多因素性质。为了解决这一差距，我们开发了PolyCheck，这是一种混合预测模型，集成了基于网络和基于规则的方法，以识别多种药物方案引起的潜在不良反应。我们构建了一个异质性的药物-靶点- adr相互作用网络，包括一线抗结核和抗糖尿病药物、它们的靶点和相关的adr。采用随机行走与重启（RWR）算法对ADR节点进行排序，并通过纳入药物-靶点-ADR关联的生物学相关性，基于规则的层进一步改进了预测。交叉验证和基于案例的检验显示了较强的预测性能，准确率、精密度、召回率、f1评分和AUPRC值分别为0.70、0.74、0.92、0.81和0.74。PolyCheck为预测复杂治疗方案中的不良反应提供了一种可扩展和可解释的方法，并可支持对合并症患者进行更安全、个性化的结核病治疗。

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引用次数: 0

Investigation of in vitro IKr/hERG assays under physiological temperature conditions using the semi-automated patch-clamp system QPatch compact with temperature control system 采用带温度控制系统的半自动膜片钳系统QPatch研究生理温度条件下体外IKr/hERG的测定

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.2025.107814

Kazuya Tsurudome, Hironori Ohshiro, Taku Izumi

Cardiac ion channel activity is crucial for generating cardiac action potentials with proper timing and duration. Drug-induced impairment of these ion channels can cause abnormal cardiac activity, including QT interval prolongation, ventricular arrhythmia, and, in the most severe cases, sudden death. These adverse effects are among the leading reasons for drug withdrawal from the market or the denial of regulatory approval for new therapeutic candidates. The ICH E14/S7B Q&A released in August 2022 provided recommended conditions for best practices for in vitro assay of IKr/hERG to maintain reproducibility and consistency in evaluations. These recommendations include testing under physiological temperature conditions, as well as considering factors such as voltage protocols. In this study, we have investigated whole-cell patch-clamp measurements of hERG currents under physiological temperature conditions (36–37 °C) using the semi-automated patch-clamp system QPatch compact using the recommended best practices. Whole-cell patch-clamp recordings in hERG channel-expressing cells were performed using the QPatch Compact automated patch-clamp system with a temperature control system and the voltage protocol recommended by the CiPA project. Compared to room temperature conditions, the rise time of the hERG current was shorter and its amplitude larger under physiological temperature conditions. The tail current decay rate was also slower. The overall duration of the current was prolonged. These findings imply that temperature influences the dynamics of hERG channels, providing a more accurate reproduction of their physiological function. Furthermore, when testing the temperature-dependent effects of erythromycin, the current inhibition rate at the highest applied concentration of 1000 μM was around 50 % under room temperature conditions (25 °C). In contrast, under physiological temperature conditions, the IC50 was approximately 60 μM, and a nearly complete blockade of hERG currents was achieved at 1000 μM. This result confirms that the inhibitory effect of erythromycin is more pronounced under physiological temperature conditions. Additionally, we have tested other reference compounds, such as dofetilide, ondansetron, and moxifloxacin, to assess their temperature sensitivity. These insights are expected to improve our understanding of the influence of temperature on drug effects and enhance the reliability of testing protocols in accordance with ICH guidelines.

心脏离子通道的活性是产生适当的时间和持续时间的心脏动作电位的关键。药物引起的这些离子通道损伤可引起心脏异常活动，包括QT间期延长、室性心律失常，在最严重的情况下可导致猝死。这些不良反应是药物退出市场或监管部门拒绝批准新的候选治疗药物的主要原因之一。2022年8月发布的ICH E14/S7B Q&；A为IKr/hERG体外测定的最佳实践提供了推荐条件，以保持评估的可重复性和一致性。这些建议包括在生理温度条件下进行测试，以及考虑电压协议等因素。在这项研究中，我们研究了在生理温度条件下（36-37 °C）使用半自动膜片钳系统QPatch compact的全细胞膜片钳测量hERG电流，并采用推荐的最佳实践。在表达hERG通道的细胞中，使用带有温度控制系统和CiPA项目推荐的电压方案的QPatch Compact自动膜片钳系统进行全细胞膜片钳记录。与室温条件相比，生理温度条件下hERG电流的上升时间更短，幅度更大。尾电流衰减速率也较慢。电流的总持续时间延长了。这些发现表明，温度会影响hERG通道的动力学，从而更准确地再现其生理功能。此外，当测试红霉素的温度依赖性时，在室温条件下（25 °C），最高施加浓度为1000 μM时，当前的抑制率约为50 %。相比之下，在生理温度条件下，IC50约为60 μM，在1000 μM时几乎完全阻断了hERG电流。这一结果证实了红霉素在生理温度条件下的抑制作用更为明显。此外，我们还测试了其他参比化合物，如多非利特、昂丹司琼和莫西沙星，以评估它们的温度敏感性。这些见解有望提高我们对温度对药物效应影响的理解，并根据ICH指南提高检测方案的可靠性。

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引用次数: 0

Compound A, a novel dual mu and kappa opioid receptor agonist that exerts potent an analgesic effect comparable to oxycodone, showed no reinforcing effect in monkeys 化合物A是一种新型的双mu和kappa阿片受体激动剂，具有与羟考酮相当的镇痛作用，但在猴子身上没有增强作用

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.2025.107800

Yukiko Orita , Atsushi Nakamura , Yuki Azuma , Kana Yasufuku , Erika Kasai , Tohko Arai

Opioids (opioid mu receptor [MOR] agonists) are one of the most powerful analgesics and are widely used around the world, however, the rapid increase in the number of deaths due to drug abuse and overdose have become a major social problem. One of the causes of opioid-mediated abuse is known to be an increase of dopamine release in nucleus accumbens (NAc) via MOR activation in brain, and this effect is also known to be counteracted by opioid kappa receptor (KOR) activation expressed in NAc. We, therefore, hypothesized that a MOR/KOR dual agonist would be able to avoid the risk of abuse and psychological dependence while maintaining a strong analgesic effect. In this study, we evaluated the analgesic effects and reinforcing effect of Compound A, a novel MOR/KOR dual agonist (non-morphinan structure) that we created and compared with those of oxycodone. To investigate in vitro functional activity of compound A, cAMP assay was conducted. Hot-plate test (55°C) in rats (n = 5/dose, p.o) was performed to assess the analgesic effect. The time-course of dopamine level in NAc in rats was determined using in vivo microdialysis method (n = 3–8/dose, i.v.). Abuse liability was assessed by rat conditioned place preference (CPP) test (n = 7/dose, i.v.) and monkey intravenous self-administration study under a fixed-ratio 30 schedule of reinforcement (n = 4/dose, i.v.). In vitro profile, Compound A exhibited MOR and KOR agonism. Compound A exhibited dose-dependent analgesic effect, with a maximum response comparable to or greater than that of oxycodone. Compound A introduced less level dopamine release in rat NAc compared to oxycodone and no increase of CPP score at the analgesic dose in rats. In monkeys, Compound A did not show any reinforcing properties over a wide dose range covering the expected clinical pharmacological exposure, whereas oxycodone showed a strong reinforcing effect. MOR/KOR dual agonists may be able to overcome the risk of abuse and psychological dependence of traditional opioids while maintaining a potent analgesic effect comparable to them, which would contribute to one solution of current opioid crisis.

阿片受体激动剂（opioid mu receptor [MOR] agonists）是最有效的镇痛药之一，在世界范围内被广泛使用，然而，由于药物滥用和过量而导致的死亡人数迅速增加已成为一个主要的社会问题。阿片介导的滥用的原因之一是通过大脑MOR激活增加伏隔核（NAc）的多巴胺释放，并且这种作用也被NAc中表达的阿片受体（KOR）激活所抵消。因此，我们假设MOR/KOR双重激动剂能够避免滥用风险和心理依赖，同时保持强大的镇痛效果。在这项研究中，我们评估了化合物A的镇痛作用和强化作用，化合物A是我们创造的一种新的MOR/KOR双激动剂（非吗啡肽结构），并与羟考酮进行了比较。为考察化合物A的体外功能活性，采用cAMP测定法。采用大鼠热板实验（55°C）（n = 5只/剂，p.o）评价其镇痛作用。采用体内微透析法测定大鼠NAc中多巴胺水平的时程（n = 3-8 /剂量，静脉注射）。采用大鼠条件位置偏好（CPP）试验（n = 7/剂量，i.v.）和猴静脉内自给药研究（n = 4/剂量，i.v.）评估滥用倾向。体外分析显示，化合物A具有MOR和KOR激动作用。 化合物A表现出剂量依赖性镇痛作用，最大反应可与羟考酮相当或更大。与羟考酮相比，化合物A在大鼠NAc中引起的多巴胺释放水平较低，且在镇痛剂量下大鼠CPP评分未增加。在猴子中，化合物A在覆盖预期临床药理学暴露的大剂量范围内没有表现出任何强化特性，而羟考酮则表现出很强的强化作用。MOR/KOR双激动剂可能能够克服传统阿片类药物滥用和心理依赖的风险，同时保持与之相当的强效镇痛效果，这将有助于解决当前阿片类药物危机。

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