Anticonvulsant screening using chronic epilepsy models

使用慢性癫痫模型进行抗惊厥筛查

• 批准号: 9316238
• 负责人: YEVGENY BERDICHEVSKY
• 金额: $ 44.39万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9316238
• 关键词: Acute; Animal Model; Animals; Anticonvulsants; Antiepileptogenic; Biochemical; Biological Assay; Biological Markers; Blinded; Blood - brain barrier anatomy; Brain; Chronic; Clinical; Clinical Trials; Collection; Convulsants; Coxibs; Cross-Over Trials; Custom; Cyclooxygenase Inhibitors; Data; Detection; Development; Dose; Double-Blind Method; Epilepsy; Epileptogenesis; Frequencies; Funding Mechanisms; Generic Drugs; Goals; Hippocampus (Brain); In Vitro; Injury; Knock-out; Knockout Mice; Laboratories; Lead; Medical; Methodology; Modeling; Monitor; National Institute of Neurological Disorders and Stroke; Neurology; Oral; PTGS2 gene; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phase; Positioning Attribute; Preclinical Drug Evaluation; Preparation; Process; Property; Protocols documentation; Publications; Publishing; Random Allocation; Randomized; Recurrence; Reproducibility; Research Personnel; Resources; Seizures; Site; Slice; Supervision; System; Technology; Telemetry; Testing; Therapeutic; United States National Institutes of Health; Vertebral column; base; brain tissue; cardiovascular risk factor; celecoxib; computerized; improved; in vitro Assay; in vitro Model; in vivo; in vivo Model; kainate; novel; programs; response; screening; synaptogenesis

Abstract The NINDS Anticonvulsant Screening Program (ASP) has identified most of the anticonvulsants in clinical use today. However, one third of epileptic patients do not respond to these drugs. The ASP protocols are based on seizures induced by subjecting normal animals to acute convulsant conditions. We have developed a complimentary system of novel in vitro and in vivo assays of spontaneous seizures in chronically epileptic preparations. This two-stage screening system provides a unique focus on recurrent spontaneous seizures in chronic epilepsy models. The first stage is an in vitro assay comprised of the organotypic hippocampal slice culture, which develops electrographic seizure activity and corresponding biochemical biomarkers over the first week in vitro. The second stage is an in vivo assay comprised of the kainate model of epilepsy in which spontaneous seizures are monitored using continuous telemetry and supervised, blinded, computerized seizure detection. We used the rapid in vitro assay to screen over 400 compound-concentration combinations from the NINDS Custom Compound Collection. We found a lead compound, celecoxib, and then verified this lead by the second-stage testing in a randomized double blind in vivo crossover trial. Celecoxib had no effect on seizures induced by acute application of convulsants to normal brain tissue, suggesting that its anticonvulsant properties are unique to chronic epilepsy, and raising the possibility that its spectrum of action will be distinct from anticonvulsants discovered by the ASP protocols. The next step in development is medicinal chemistry to optimize celecoxib’s anticonvulsant efficacy. This is most feasibly accomplished through the UH2 / UH3 Blueprint Neurotherapeutics Network. As our discussions with BPN program officers clarified, to efficiently utilize the BPN medicinal chemistry program we must further develop the in vitro and in vivo assays and acquire additional data on our lead compound. The UH2/3 mechanism was considered the most appropriate funding mechanism by the NINDS program officer. In the R21 phase of this proposal, we will extend the in vitro assay’s concentration-response for celecoxib and 2,5 dimethyl celecoxib, a derivative that does not inhibit COX2 but has equal anticonvulsant efficacy in vitro. We will then characterize the assay’s reproducibility and Z factor. We will also establish the dose-response of the in vivo assays for celecoxib, and increase the number of in vitro and in vivo sites to two each in order to improve robustness and throughput, as well as engage outstanding younger investigators in this effort. In the R33 phase of the proposal, we will further characterize the lead compound by determining whether COX2 inhibition is necessary for anticonvulsant activity in the in vitro and in vivo assays.

抽象的 NINDS 抗惊厥药筛选计划 (ASP) 已鉴定出大多数临床使用的抗惊厥药 今天。然而，三分之一的癫痫患者对这些药物没有反应。 ASP 协议基于 对正常动物进行急性惊厥引起的癫痫发作。我们开发了一个 慢性癫痫患者自发性癫痫发作的新型体外和体内检测互补系统 准备工作。这种两阶段筛查系统特别关注复发性自发性癫痫发作 慢性癫痫模型。第一阶段是体外测定，包括器官型海马切片 培养，在第一代中形成电图癫痫发作活动和相应的生化生物标志物 体外一周。第二阶段是由红藻氨酸癫痫模型组成的体内测定，其中 使用连续遥测技术监测自发性癫痫发作，并进行监督、盲法、计算机化 癫痫发作检测。我们使用快速体外测定筛选了 400 多种化合物浓度组合 来自 NINDS 定制化合物集合。我们发现了一种先导化合物塞来昔布，然后验证了这一点 由随机双盲体内交叉试验的第二阶段测试领导。塞来昔布没有效果 对正常脑组织急性使用惊厥剂引起的癫痫发作的研究表明， 抗惊厥特性是慢性癫痫所独有的，并且增加了其作用范围的可能性 与 ASP 协议发现的抗惊厥药不同。 开发的下一步是药物化学，以优化塞来昔布的抗惊厥功效。这是 最可行的是通过 UH2 / UH3 蓝图神经治疗网络来完成。正如我们的讨论 BPN 项目官员澄清，为了有效利用 BPN 药物化学项目，我们必须进一步 开发体外和体内测定并获取有关我们的先导化合物的额外数据。 UH2/3 NINDS 项目官员认为该机制是最合适的筹资机制。在 该提案的 R21 阶段，我们将扩展塞来昔布和 2,5 的体外测定浓度响应 二甲基塞来昔布是一种衍生物，不抑制COX2，但在体外具有同等的抗惊厥功效。我们 然后将表征测定的重现性和 Z 因子。我们还将建立剂量反应 塞来昔布的体内测定，并将体外和体内位点的数量各增加到两个，以便 提高稳健性和吞吐量，并吸引优秀的年轻研究人员参与这项工作。在 在提案的 R33 阶段，我们将通过确定 COX2 是否存在来进一步表征先导化合物 抑制对于体外和体内试验中的抗惊厥活性是必要的。