Validation of High-Throughput Screening of Candidate Antiepileptogenic Drugs

描述（由申请人提供）：了解慢性癫痫（即癫痫发生）演变的机制有助于开发有效的癫痫过程预防疗法。反过来，需要开发和验证临床相关的动物模型，这些模型可以用于基础和临床前研究。目前大多数可用的癫痫发生模型采用诱发性损伤（如癫痫持续状态，SE），导致进行性神经退行性和神经元可塑性改变，最终导致自发性复发性癫痫发作。虽然这些模型是基础癫痫研究不可或缺的一部分，但由于时间、劳动力和成本的原因，这些模型的固有特征（如进展缓慢、癫痫发作频率和严重程度在动物间和时间上的高度可变性、个体癫痫发作的不可预测性）对临床前抗癫痫药物（AED）评估构成了重大挑战。拟议项目的目的是验证一种高通量筛选试验，用于快速识别和评估抗癫痫和/或疾病改善药物。我们认为，癫痫发生的快速点燃模型可用于AED药物的开发。验证将包括分析15种具有既定药理学特征的化合物（布美他尼、地西泮、非氨基甲酸乙酯、加巴喷丁、加那索龙、拉莫三嗪、左乙拉西坦、MK-801、苯妥英、瑞加滨、托吡酯、丙戊酸钠、维加巴林、唑尼沙胺）对5个不同年龄（新生儿、新生儿后期、青春期前、青春期、成年）实验动物快速点燃癫痫发生的影响。每5分钟以超阈电流向海马体施加60次电刺激（总过程持续时间为5小时），可诱发快速点火。动物将在点火前接受一次AED注射；电图和行为癫痫的进展和模式将被监测和分析。选择aed的目的是，基于其作用机制和已知的癫痫发生机制，它们有望以不同的方式改变癫痫发作的进展，包括完全预防癫痫发作（抗癫痫作用），减轻癫痫过程（疾病改变），以及根本不影响癫痫发作。作为研究的结果，我们将产生一个扩大的药理学概况的快速点燃模型的癫痫发生。编译后的数据库可用于绘制新出现的抗癫痫药物与已知抗癫痫药的疗效对比图。癫痫的进行性（癫痫发生）决定了开发能够在早期阶段阻断癫痫过程的治疗方法的必要性。反过来，需要引入和验证实验模型，这些模型将(a)与人类状况非常相似，(b)允许大规模筛选前瞻性抗癫痫药物。目前的项目旨在引入和验证癫痫发生的动物模型，该模型将允许快速有效地大规模筛选新兴的抗癫痫药物。

DESCRIPTION (provided by applicant): Understanding mechanisms that underlie the evolution of chronic epilepsy (i.e. epileptogenesis) is instrumental for developing effective preventive therapies of the epileptic process. In turn, the development and validation of clinically relevant animal models are needed that could be used for both basic and preclinical studies. The majority of currently available models of epileptogenesis employ a precipitating insult (e.g. status epilepticus, SE), which leads to progressive neurodegenerative and neuronal plastic changes, and culminates in the occurrence of spontaneous recurrent seizures. While being indispensable for basic epilepsy research, the inherent features of these models (e.g. slow progression; high inter-animal and temporal variability of seizure frequency and severity; unpredictability of individual seizure occurrence) represent significant challenges for preclinical antiepileptic drug (AED) evaluation due to time, labor and costs involved. The objective of the proposed project is to validate a high-throughput screening assay for rapid identification and evaluation of antiepileptogenic and/or disease-modifying drugs. We propose that the rapid kindling model of epileptogenesis can be used for AED drug discovery. The validation will consist of analysis of the effects of fifteen compounds with established pharmacological profiles (bumetanide, diazepam, ethosuximide felbamate, gabapentin, ganaxolone, lamotrigine, levetiracetam, MK-801; phenytoin, retigabine, topiramate, valproate sodium, vigabatrin, zonisamide) on rapid kindling epileptogenesis in experimental animals of five different ages (neonatal; post-neonatal; pre-adolescent; adolescent; adult). Rapid kindling will be induced by 60 electrical stimulations delivered to the hippocampus every 5 minutes at a suprathreshold current (total procedure duration is 5 hours). The animals will receive a single injection of an AED prior to kindling; the progression and the pattern of electrographic and behavioral seizures will be monitored and analyzed. AEDs are chosen so that, based on the mechanisms of their action and known mechanisms of epileptogenesis, they are expected to modify seizure progression in different ways, including complete prevention of seizures (antiepileptogenic effect), mitigation of epileptic process (disease modification), and failure to affect seizures at all. As a result of the study, we will generate an expanded pharmacological profile of the rapid kindling model of epileptogenesis. The compiled database can then be used for mapping of emerging antiepileptogenic drugs against known AEDs in terms of their efficiency. The progressive nature of epilepsy (epileptogenesis) dictates the necessity for developing therapies that can block the epileptic process during its early stages. In turn, the introduction and validation of experimental models that would (a) closely resemble human condition, and (b) allow large scale screening of prospective antiepileptogenic drugs are required. The current project is designed to introduce and validate an animal model of epileptogenesis that would allow for the quick and effective screening of emerging antiepileptogenic drugs on a large scale.