Developing a selective TRPC3 ion channel inhibitor for epilepsy treatment

项目摘要 癫痫是最常见的脑部疾病之一。目前的药物疗效有限。确定新的 可以作为更安全、更有效治疗方法的靶向药物正处于迫切的未得到满足的需求中。选择性抑制 作为一种阻止癫痫发作的新策略，TRPC3应运而生。 然而，目前最好的选择性TRPC3抑制剂，PYR3，代谢稳定性差，具有显著的 安全责任。我们最近公布了一种专利的Pyr3类似物JW-65的发现，它具有显著的 改善代谢稳定性和安全性。它具有良好的脑渗透性和滞留性，直接结合到 TRPC3，在癫痫的头对头比较中显示出比现有药物苯妥英更好的疗效 老鼠模型。Seak正在授权这种专利脚手架，并建议在第一阶段STTR中彻底降低风险 JW-65作为靶向癫痫治疗的潜在临床候选药物。 目标1.确定毒物学特征，针对一组生理指标确定潜在的脱靶 重要靶点，主要的CyP抑制和诱导，膜通透性，转运蛋白效应， 血浆结合和稳定性，以及药代动力学(PK)曲线，以进一步降低JW-65的风险。我们已经这么做了 证实了JW-65与TRPC3的直接结合和功能抑制，并展示了其优良的药物-- 就像房产一样。我们还表明JW-65与I.P.具有良好的PK性能。注射。为了实现这一目标，我们将 首先进行JW-65的大规模合成，以支持后续的生物学评价。到时候我们会的 综合评价其安全性、脱靶、ADME特性和行业标准RAT PK(i.v.和口头) 全面降低JW-65作为临床候选病例的风险。里程碑：(1)三克JW-65 合成并严格表征；(2)证明JW-65的安全性和类药物特性 作为一个可行的候选者；(3)测定JW-65的PK谱和口服生物利用度。 目的2.评价JW-65对多种模型急性癫痫发作的抑制作用 在老鼠和大鼠身上都是如此。我们将首先评估JW-65对匹罗卡品诱导的大鼠急性癫痫的疗效， 同时使用PYR3和苯二氮卓类药物(延长癫痫发作的一线药物)作为参考。为了进一步 增加严格性并避免任何型号或物种特定的发现，JW-65的抗癫痫效果将是 在小鼠红藻氨酸模型和氟乙基模型中得到验证。里程碑：(4)展示对急性呼吸道感染的疗效 匹罗卡品大鼠癫痫发作(行为癫痫评分或EEG尖峰降低约50%)；(5)显示 对氟乙基诱发的小鼠癫痫发作的疗效(首次肌阵挛发作的潜伏期增加~50%，并 全身性强直-阵挛发作)；(6)对6赫兹诱发的小鼠癫痫发作(~50%)有疗效 行为癫痫评分下降)；(7)表现出对自发性癫痫发作的疗效(~50%下降 每天的SRS总数)。 未来方向：在第二阶段，Seak将为JW-65进行全面的IND使能研究。

Project Summary Epilepsy is one of the most common brain disorders. Current drugs have limited efficacy. Identifying new targeted drugs that can be used as safer, more effective therapies is in urgent unmet need. Selective inhibition of the transient receptor potential canonical 3 (TRPC3) emerges as a novel strategy to impede epilepsy. However, the currently best selective TRPC3 inhibitor, Pyr3, has poor metabolic stability and has significant safety liabilities. We recently published the discovery of a patented Pyr3 analog, JW-65, which has significantly improved metabolic stability and safety profiles. It has good brain penetration and retention, directly binds to TRPC3, and shows better efficacy than an existing drug phenytoin in a head-to-head comparison in an epilepsy mouse model. SEAK is licensing this patented scaffold and proposes in this Phase I STTR to thoroughly de-risk JW-65 as a potentially viable clinical candidate for targeted epilepsy therapy. Aim 1. Determine toxicological profiles, potential off-targets against a panel of physiologically important targets, major CYP inhibitions and inductions, membrane permeability, transporter effects, plasma binding and stability, and pharmacokinetic (PK) profiles to further de-risk JW-65. We have already confirmed the direct binding to and functional inhibition of TRPC3 by JW-65 and demonstrated its excellent drug- like properties. We have also showed that JW-65 has good PK properties with i.p. injection. In this aim, we will first perform large scale synthesis of JW-65 to support subsequent biological evaluations. We will then comprehensively evaluate its safety, off-targets, ADME properties, and industry standard rat PK (i.v. and oral) profiles to comprehensively de-risk JW-65 as a clinical candidate. Milestones: (1) three grams of JW-65 synthesized and rigorously characterized; (2) demonstrate the safety profiles and drug like properties of JW-65 as a viable candidate; (3) determine PK profiles and oral bioavailability of JW-65. Aim 2. Evaluate in vivo efficacy of JW-65 for its ability to suppress acute seizures in multiple models in both mice and rats. We will first assess the efficacy of JW-65 on acute seizures in rats induced by pilocarpine, using both Pyr3 and benzodiazepines (the first-line drugs for prolonged seizures) as references. To further increase the rigor and avoid any model- or species-specific findings, the anti-seizure effects of JW-65 will be validated in the mouse kainic acid model and flurothyl model. Milestones: (4) demonstrate efficacy against acute pilocarpine seizures in rats (~50% reduction in behavioral seizure scores or EEG spikes); (5) demonstrate efficacy against flurothyl-induced seizures in mice (~50% increase in latencies to the first myoclonic jerk and generalized tonic-clonic seizure); (6) demonstrate efficacy against 6 Hz-induced seizures in mice (~50% decrease in behavioral seizure scores); (7) demonstrate efficacy against spontaneous seizures (~50% decrease in total number of SRSs per day). Future directions: In Phase 2, SEAK will perform comprehensive IND-enabling studies for JW-65.