Optimization of the adult zebrafish ECG method for assessment of drug-induced QTc prolongation
Optimization of the adult zebrafish ECG method for assessment of drug-induced QTc prolongation
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DOI：
10.1016/j.vascn.2013.01.007
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发表时间：
2013-03-01
影响因子：
1.9
通讯作者：
Kulkarni, Pushkar
中科院分区：
文献类型：
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作者：
Chaudhari, Girish Hari;Chennubhotla, Keerthana Sarma;Kulkarni, Pushkar
简介：最近的研究表明，成年斑马鱼心电图（心电图）在评估药物引起的 QTc 延长方面很有用。虽然该方法比当前的心电图动物模型具有显着优势，包括伦理问题、低化合物需求和费用，但该方法在药物发现项目中的采用受到了特定限制的阻碍。局限性包括无法确定所施用的测试化合物的确切剂量，以及由于口腔灌注流速和固定方法等变量造成的潜在影响。我们描述了一种可重复记录成年斑马鱼心电图的改进方法，并说明了其在使用组胺受体拮抗剂特非那定作为测试药物研究药物诱导的 QTc 延长中的应用。方法：以成年斑马鱼每公斤体重的毫克数为基础，选择肠外给药而非灌注。优化适应和固定方法，以避免由于突然的环境变化而导致心电图伪影。我们进一步修改了 QT 校正的公式，并确保稳定心电图的可重复记录。使用不同浓度的特非那丁，并与基线和未治疗的对照相比，分析所产生的致心律失常效应。结果：所有斑马鱼均记录到正常、稳定且可重复的心电图。发现0.1 mg/kg体重的特非那定为NOAEL。我们发现，在所有浓度下，已知的人类 QTc 效应与在成年斑马鱼中观察到的 QTc 效应之间存在极好的相关性。在斑马鱼中观察到的所有特非那定诱导的致心律失常作用均具有剂量和时间依赖性。讨论：我们报告了一种可重复记录稳定斑马鱼心电图的改进方法，以促进其在延长 QTc 药物的临床前研究中的常规应用，并可靠估计 NOAEL。我们的研究与药物发现中替代动物模型的开发和使用相关。 (C) 2013 Elsevier Inc. 保留所有权利。
Introduction: Recent studies have shown the utility of adult zebrafish ECG (electrocardiogram) in assessing drug-induced QTc prolongation. While the method has significant advantages over current ECG animal models including ethical issues, low compound requirement and expense, adoption of the method into drug discovery programs has been hampered by specific limitations. The limitations include the inability to determine the exact dose of test compound administered, and potential effects due to variables such as flow rate of oral perfusion and immobilization method. We describe a refined method for the reproducible recording of the adult zebrafish ECG and illustrate its application in investigating drug-induced QTc prolongation using the histamine receptor antagonist Terfenadine as a test drug. Method: We chose to perform parenteral administration of test drug instead of perfusion on the basis of mg per kg body weight of adult zebrafish. Acclimatization and immobilization methods were optimized to avoid ECG artifacts due to sudden environmental changes. We further modified the formula for QT correction and ensured reproducible recording of stable ECGs. Various concentrations of Terfenadine were used and the resultant proarrhythmic effects were analyzed as compared to the baseline and untreated controls. Results: Normal, stable and reproducible ECGs were recorded in all zebrafish. Terfenadine at the rate of 0.1 mg/kg body weight was found to be the NOAEL. We found an excellent correlation between known QTc effects in humans and those observed in adult zebrafish at all concentrations. All Terfenadine-induced proarrhythmic effects observed in zebrafish were dose and time dependent. Discussion: We report a refined method for reproducible recording of stable zebrafish ECGs to facilitate its routine application in preclinical investigation of QTc-prolonging drugs with reliable estimation of NOAEL. Our study is of relevance to the development and use of alternate animal models in drug discovery. (C) 2013 Elsevier Inc. All rights reserved.