DataSheet1_Zebrafish EEG predicts the efficacy of antiepileptic drugs.pdf

Background: Pharmacological evaluation of antiepileptic drugs (AEDs) using mammalian animals takes long time and is expensive. The zebrafish is a species commonly used to study brain functions, neurological diseases, and drug toxicity, and attracts more attention as an alternative animal model to substitute or supplement mammalian animals in drug development. Electroencephalogram (EEG) is a key indicator for diagnosing brain diseases such as epilepsy, by directly measuring the brain activity. We propose a novel method for pharmacological evaluation of AEDs based on EEG from adult zebrafish, which allows researchers to select more clinically valuable drugs at the early stage of AED screening.
Methods: To evaluate the efficacy of AEDs, zebrafish EEG signals were measured after administering six AEDs (valproate acid, gabapentin, ethosuximide, oxcarbazepine, tiagabine, and topiramate) at various doses to pentylenetetrazol (PTZ)-induced seizure models. The change in seizure activity was investigated according to doses. The antiepileptic effect was determined by observing a significant decrease in at least one out of three indicators of the number, total duration, and mean duration of ictal events.
Results: Using EEG signals from adult zebrafish, antiepileptic effects were observed with all six AEDs. Among them, antiepileptic effects depending on dose were confirmed with valproate acid, gabapentin, ethosuximide, and tiagabine. Moreover, the 50% effective doses (ED50) of valproate acid and tiagabine were determined based on zebrafish EEG for the first time, indicating that the quantitative inter-species comparison of the AED efficacy is possible between zebrafish and mammals such as rodents.
Significance: The results show that zebrafish can be used to effectively and quantitatively evaluate the efficacy of AEDs based on EEG, the same method to evaluate antiepileptic effects in mammals, suggesting that the proposed method can contribute in reducing the cost and duration of search for AEDs and thus accelerate the drug development cycles.

研究背景：采用哺乳动物开展抗癫痫药物（antiepileptic drugs, AEDs）的药理学评价，不仅耗时冗长，且成本高昂。斑马鱼是一类常用于脑功能、神经系统疾病及药物毒性研究的模式生物，作为药物研发中替代或补充哺乳动物的动物模型，其关注度与日俱增。脑电图（electroencephalogram, EEG）通过直接检测脑电活动，是癫痫等脑部疾病诊断的关键指标。本研究提出一种基于成年斑马鱼脑电图的抗癫痫药物药理学评价新方法，可助力研究者在抗癫痫药物筛选的早期阶段，遴选更具临床价值的候选药物。
研究方法：为评估抗癫痫药物的疗效，本研究对戊四氮（pentylenetetrazol, PTZ）诱导的癫痫模型斑马鱼，以不同剂量给药6种抗癫痫药物（丙戊酸、加巴喷丁、乙琥胺、奥卡西平、替加宾、托吡酯），随后检测其脑电信号。研究人员根据给药剂量探究癫痫发作活动的变化规律，并以发作事件的数量、总持续时长及平均持续时长三项指标中至少一项出现显著降低，作为抗癫痫疗效的判定标准。
研究结果：基于成年斑马鱼脑电信号的实验显示，6种受试抗癫痫药物均表现出明确的抗癫痫疗效。其中，丙戊酸、加巴喷丁、乙琥胺及替加宾的抗癫痫疗效呈现显著的剂量依赖性。此外，本研究首次基于斑马鱼脑电信号测定了丙戊酸与替加宾的半数有效剂量（50% effective doses, ED50），证实可在斑马鱼与啮齿类等哺乳动物之间，实现抗癫痫药物疗效的跨物种定量比较。
研究意义：本研究结果表明，斑马鱼可依托脑电图技术，采用与哺乳动物体内抗癫痫疗效评价一致的逻辑，实现抗癫痫药物疗效的有效定量评价。该方法可有效降低抗癫痫药物研发的成本与周期，进而加速药物研发进程。