Griffin Aliesha, Anvar Mana, Hamling Kyla, Baraban Scott C
Epilepsy Research Laboratory and Weill Institute for Neuroscience, Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, United States.
Front Pharmacol. 2020 Apr 24;11:464. doi: 10.3389/fphar.2020.00464. eCollection 2020.
Dravet syndrome is a catastrophic epilepsy of childhood, characterized by cognitive impairment, severe seizures, and increased risk for sudden unexplained death in epilepsy (SUDEP). Although refractory to conventional antiepileptic drugs, emerging preclinical and clinical evidence suggests that modulation of the endocannabinoid system could be therapeutic in these patients. Preclinical research on this topic is limited as cannabis, delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD), are designated by United States Drug Enforcement Agency (DEA) as illegal substances. In this study, we used a validated zebrafish model of Dravet syndrome, scn1lab homozygous mutants, to screen for anti-seizure activity in a commercially available library containing 370 synthetic cannabinoid (SC) compounds. SCs are intended for experimental use and not restricted by DEA designations. Primary phenotype-based screening was performed using a locomotion-based assay in 96-well plates, and a secondary local field potential recording assay was then used to confirm suppression of electrographic epileptiform events. Identified SCs with anti-seizure activity, in both assays, included five SCs structurally classified as indole-based cannabinoids JWH 018 N-(5-chloropentyl) analog, JWH 018 N-(2-methylbutyl) isomer, 5-fluoro PB-22 5-hydroxyisoquinoline isomer, 5-fluoro ADBICA, and AB-FUBINACA 3-fluorobenzyl isomer. Our approach demonstrates that two-stage phenotype-based screening in a zebrafish model of Dravet syndrome successfully identifies SCs with anti-seizure activity.
德拉韦特综合征是一种儿童期灾难性癫痫，其特征为认知障碍、严重癫痫发作以及癫痫性猝死（SUDEP）风险增加。尽管对传统抗癫痫药物难治，但新出现的临床前和临床证据表明，调节内源性大麻素系统可能对这些患者具有治疗作用。由于大麻、Δ⁹-四氢大麻酚（THC）和大麻二酚（CBD）被美国药物管制局（DEA）指定为非法物质，关于这一主题的临床前研究有限。在本研究中，我们使用经过验证的德拉韦特综合征斑马鱼模型——scn1lab纯合突变体，在一个包含370种合成大麻素（SC）化合物的商业文库中筛选抗癫痫活性。SCs intended for experimental use and not restricted by DEA designations. 使用基于运动的96孔板检测法进行基于主要表型的筛选，然后使用二次局部场电位记录检测法来确认对脑电图癫痫样事件的抑制。在两种检测中均具有抗癫痫活性的已鉴定SCs包括五种结构上归类为吲哚类大麻素的SCs：JWH 018 N-（5-氯戊基）类似物、JWH 018 N-（2-甲基丁基）异构体、5-氟PB-22 5-羟基异喹啉异构体、5-氟ADBICA和AB-FUBINACA 3-氟苄基异构体。我们的方法表明，在德拉韦特综合征斑马鱼模型中基于表型的两阶段筛选成功鉴定出具有抗癫痫活性的SCs。
