Validation of High-Throughput Screening of Candidate Antiepileptogenic Drugs

描述（由申请人提供）：了解慢性癫痫演变（即癫痫发生）的机制有助于开发癫痫过程的有效预防性治疗。反过来，需要开发和验证可用于基础和临床前研究的临床相关动物模型。目前可用的大多数癫痫发生模型采用了一种诱发性损伤（例如癫痫持续状态，SE），导致进行性神经退行性和神经元可塑性变化，并最终导致自发性复发性癫痫发作的发生。虽然这些模型对于基础癫痫研究是不可或缺的，但其固有特征（例如进展缓慢;癫痫发作频率和严重程度的动物间和时间变异性高;个体癫痫发作发生的不可预测性）代表了临床前抗癫痫药物（AED）评价的重大挑战，这是由于所涉及的时间，劳动力和成本。该项目的目的是验证一种用于快速鉴定和评价抗癫痫和/或疾病修饰药物的高通量筛选方法。我们建议癫痫发生的快速点燃模型可用于AED药物的发现。验证将包括分析具有已确定药理学特征的15种化合物（布美他尼、地西泮、乙琥胺、非氨酯、加巴喷丁、加奈索酮、拉莫三嗪、左乙拉西坦、MK-801;苯妥英、瑞替加滨、托吡酯、丙戊酸钠、氨己烯酸、唑尼沙胺）对5种不同年龄（新生儿、新生儿后、青春期前、青春期、成年）实验动物快速点燃癫痫发生的影响。通过以阈上电流每5分钟向海马体输送60次电刺激诱导快速点燃（总手术持续时间为5小时）。动物将在点燃前接受AED单次注射;将监测和分析电图和行为癫痫发作的进展和模式。选择AED，以便基于其作用机制和已知的癫痫发生机制，预期其以不同方式改变癫痫发作进展，包括完全预防癫痫发作（抗癫痫作用）、缓解癫痫过程（疾病改善）和根本不影响癫痫发作。作为这项研究的结果，我们将产生一个扩大的药理学概况的快速点燃模型癫痫。编译的数据库，然后可以用于映射新兴的抗癫痫药物对已知的抗癫痫药物的效率。癫痫的进行性（癫痫发生）决定了开发可以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）非常类似于人类状况，并且（B）允许大规模筛选有前景的抗癫痫药物。目前的项目旨在介绍和验证癫痫发生的动物模型，这将允许快速有效地筛选新兴的抗癫痫药物的大规模。

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.