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 可用作更安全、更有效疗法的靶向药物迫切需要得到满足。选择性抑制 瞬时受体电位典范3（TRPC 3）的出现作为一种新的策略，以阻止癫痫。 然而，目前最好的选择性TRPC 3抑制剂Pyr 3具有差的代谢稳定性，并且具有显著的生物学活性。 安全责任。我们最近发表了一项专利Pyr 3类似物JW-65的发现， 改善代谢稳定性和安全性。它具有良好的大脑渗透和保留，直接结合到 TRPC 3，在癫痫患者的头对头比较中显示出比现有药物苯妥英更好的疗效。 小鼠模型SEAK正在许可这种专利支架，并在第一阶段STTR中建议彻底消除风险 JW-65作为靶向癫痫治疗的潜在可行临床候选药物。 目标1。确定毒理学特征、一组生理学指标的潜在脱靶 重要的靶点，主要的抑制和诱导，膜通透性，转运蛋白的作用， 血浆结合和稳定性以及药代动力学（PK）特征，以进一步降低JW-65的风险。我们已经 证实了JW-65对TRPC 3的直接结合和功能性抑制，并证明了其优异的药物活性。 比如财产我们还表明JW-65在腹膜内注射时具有良好的PK特性。为此，我们会 首先进行JW-65的大规模合成，以支持后续的生物学评价。然后我们将 全面评价其安全性、脱靶、ADME特性和行业标准大鼠PK（i. v.和口服） 全面降低JW-65作为临床候选药物的风险。Milestone：（1）JW-65三克 合成并严格表征;（2）证明JW-65的安全性和药物样特性 作为可行的候选物;（3）确定JW-65的PK特征和口服生物利用度。 目标二。评价JW-65在多种模型中抑制急性癫痫发作的能力的体内功效 在小鼠和大鼠中。我们将首先评估JW-65对毛果芸香碱诱导的大鼠急性癫痫发作的疗效， 使用Pyr 3和苯二氮卓类药物（长期癫痫发作的一线药物）作为参考。进一步 增加严格性并避免任何模型或物种特异性结果，JW-65的抗癫痫作用将是 在小鼠红藻氨酸模型和氟代乙酸模型中得到验证。Mildrone：（4）证明对急性 大鼠匹鲁卡品癫痫发作（行为癫痫发作评分或EEG尖峰降低约50%）;（5）证明 在小鼠中对抗氟乙烯诱导的癫痫发作的功效（对于第一次肌阵挛性痉挛， 全身强直-阵挛性癫痫发作）;（6）证明对小鼠中6 Hz诱导的癫痫发作有效（约50% 行为性癫痫发作评分降低）;（7）证明对自发性癫痫发作有效（降低约50% 每天的SRS总数）。 未来方向：在第2阶段，SEAK将对JW-65进行全面的IND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.