Development of a therapeutic Kv7 opener to restore motor excitability in ALS

开发治疗性 Kv7 开启剂以恢复 ALS 的运动兴奋性

• 批准号: 10010630
• 负责人: Sandy Hinckley
• 金额: $ 35.76万
• 依托单位: QURALIS CORPORATION
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10010630
• 关键词: ALS patients; Adverse event; Agreement; Amyotrophic Lateral Sclerosis; Antiepileptic Agents; Autophagocytosis; Axonal Neuropathy; Biological Assay; Biological Markers; Cells; Chemical Structure; Chemicals; Clinical; Clinical Trials; Cryopreservation; Data; Development; Disease; Dose; Drug Kinetics; Drug Targeting; Electrophysiology (science); Epilepsy; Epithelial Cells; Excretory function; Exhibits; Freezing; Frequencies; Functional disorder; Future; Genetic Transcription; Goals; Industrialization; Ion Channel; Ion Channel Gating; Ions; Lead; Length; Licensing; Link; Measures; Medical; Metabolism; Methods; Motor; Motor Cortex; Motor Neurons; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Optics; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Phase; Potassium; Potassium Channel; Primary Bone Osteosarcoma; Production; Property; Protocols documentation; Reagent; Seasons; Series; Services; Spinal; Structure; Structure-Activity Relationship; System; Testing; Therapeutic; Toxic effect; Toxicology; absorption; base; counterscreen; drug development; drug discovery; drug market; electrical property; experience; experimental study; genetic manipulation; human monoclonal antibodies; in vivo; induced pluripotent stem cell; innovation; meetings; member; neuronal excitability; neurophysiology; novel; novel therapeutics; receptor; response; scaffold; scale up; screening; side effect; stem cell differentiation; stem cell model; stem cells; voltage

PROJECT SUMMARY Amyotrophic lateral sclerosis (ALS) is one of the most devastating neurodegenerative diseases and no therapies significantly extend patient lives. Thus, new therapeutics to treat ALS are needed. QurAlis is approaching this need by correcting the hyperexcitability that leads to motor axon neuropathy and neurodegeneration. This hyperexcitability is linked to deficits in the delayed-rectifier potassium (K+) current conducted by Kv7.2/7.3 voltage-gated ion channels. Approximately 50% of ALS patients have significant hyperexcitability in their motor cortex and spinal motor neurons and can be identified using neurophysiological biomarkers. Retigabine, an anti- epileptic and positive modulator of Kv7.2/7.3, has recently been shown to reverse this hyperexcitability in a clinical trial of ALS patients but unfortunately, is associated with off-target effects and is no longer available. QurAlis’s goal is to produce a more potent and specific Kv7.2/7.3 activator. In Phase I, Aim 1, we will develop a protocol for transcriptional reprogramming-based production of iMNs. In Aim 2, we will validate the iMNs cellular properties compared to iMNs programmed using standard patterning factors and will validate the electrical properties of iMNs using the OptopatchTM platform. In Aim 3, we will produce a large-scale cryopreserved bank of iMN. At Phase I conclusion, we will have achieved an iMNs purity >80% and demonstrated that these cells are electrically active and have a concentration-dependent reduction in neuronal activity in response to retigabine with an EC50 between 1 and 5 µM. In addition, we will have iMN production at a sufficient scale to perform chemical structure-activity relationship screening (e.g., frozen batches of at least 150 million cells). In Phase II, Aim 4, we will develop new chemical matter and test compounds for their ability to modulate Kv7.2/7.3 in structure-activity assays, and in Aim 5 we will identify compounds that reduce excitability in patient-derived iMNs. At conclusion of Phase II, we will have identified 10-20 compounds that meet our target product profile for on- target and off-target potency (Aim 4 milestone) and we will have two or more compounds that shift Kv7.2/7.3 activity in patient-derived iMNs toward reduced excitability at an effect size greater or equal to retigabine at its clinically efficacious dose of 2 µM. Aim 5 milestone). We have preliminary data demonstrating that our approach and goals are feasible: we have 3 independent novel chemical scaffolds that are potent activators of Kv7.2/7.3 whose activity has been validated in both heteromeric ion channels and in patient-derived MNs (traditional protocol) using the Optopatch platform. At conclusion of this study, we will have identified several lead compounds that are ready for ADME, PK, and non-GLP toxicology studies, and we will have a scaled-up protocol for producing patient-derived iMNs that will facilitate future drug development that target other pathways in ALS patients (e.g., autophagy).

项目概要 肌萎缩侧索硬化症 (ALS) 是最具破坏性的神经退行性疾病之一，目前尚无治疗方法 显着延长患者的生命。因此，需要新的疗法来治疗 ALS。 QurAlis 正在接近这个目标 通过纠正导致运动轴突神经病和神经变性的过度兴奋来满足需要。这 过度兴奋与 Kv7.2/7.3 传导的延迟整流钾 (K+) 电流的缺陷有关 电压门控离子通道。大约 50% 的 ALS 患者运动功能明显过度兴奋 皮层和脊髓运动神经元，可以使用神经生理学生物标志物进行识别。瑞替加滨，一种抗 Kv7.2/7.3 的癫痫和正调节剂，最近被证明可以逆转这种过度兴奋性 但不幸的是，针对 ALS 患者的临床试验与脱靶效应有关，因此不再可用。 QurAlis 的目标是生产更有效、更特异的 Kv7.2/7.3 激活剂。在第一阶段的目标 1 中，我们将开发 基于转录重编程的 iMN 生产协议。在目标 2 中，我们将验证 iMNs 细胞 与使用标准图案因子编程的 iMN 相比，将验证电气性能 使用 OptopatchTM 平台的 iMN 的属性。在目标 3 中，我们将生产一个大型冷冻库 iMN 的。在第一阶段结束时，我们将实现 iMNs 纯度 >80%，并证明这些细胞 具有电活性，并且响应瑞替加滨，神经元活性呈浓度依赖性降低 EC50 介于 1 至 5 µM 之间。此外，我们将拥有足够规模的 iMN 生产来执行 化学结构-活性关系筛选（例如，至少 1.5 亿个细胞的冷冻批次）。在第二阶段， 目标 4，我们将开发新的化学物质并测试化合物调节 Kv7.2/7.3 的能力 结构-活性测定，在目标 5 中，我们将鉴定可降低患者来源 iMN 兴奋性的化合物。 在第二阶段结束时，我们将鉴定出 10-20 种化合物，这些化合物符合我们的目标产品特征： 目标和脱靶效力（Aim 4 里程碑），我们将有两种或多种可改变 Kv7.2/7.3 的化合物 患者来源的 iMN 的活性降低兴奋性，其效应大小大于或等于瑞替加滨 临床有效剂量为2 µM。目标 5 里程碑）。我们有初步数据表明我们的方法 并且目标是可行的：我们拥有 3 个独立的新型化学支架，它们是 Kv7.2/7.3 的有效激活剂 其活性已在异聚离子通道和患者来源的 MN（传统 协议）使用 Optopatch 平台。在这项研究结束时，我们将确定几个主要因素 已准备好进行 ADME、PK 和非 GLP 毒理学研究的化合物，我们将有一个扩大规模的方案 用于生产源自患者的 iMN，这将促进未来针对 ALS 其他途径的药物开发 患者（例如自噬）。