Small Molecules to Promote Regeneration and Recovery Following Spinal Cord Injury

小分子促进脊髓损伤后的再生和恢复

• 批准号: 10016825
• 负责人: Samy Meroueh
• 金额: --
• 依托单位: RLR VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-10-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10016825
• 关键词: Affinity Chromatography; Animal Model; Animals; Axon; Binding; Biological Assay; Biology; Blood; Blood - brain barrier anatomy; Brain; Cell Culture Techniques; Clinical Trials; Contusions; Data; Development; Docking; Dorsal; Dose; Drug Industry; Drug Kinetics; Event; Exhibits; Fiber; Forelimb; Goals; Grant; Growth; Guanosine Triphosphate Phosphohydrolases; In Vitro; Injury; Investigational Drugs; Laboratories; Lead; Length; Lesion; Libraries; Lower Extremity; Mass Spectrum Analysis; Maximum Tolerated Dose; Mediating; Methods; Modeling; Monomeric GTP-Binding Proteins; Motor; Mus; Natural regeneration; Neurites; Neurons; Outcome; Paraplegia; Patients; Permeability; Pharmaceutical Chemistry; Positioning Attribute; Property; Proteins; Recovery; Recovery of Function; Sensory; Signal Transduction; Site; Small Molecule Chemical Library; Spinal Cord Contusions; Spinal cord injury; Spinal cord injury patients; Structural Protein; Structure; Testing; Therapeutic Agents; United States; aggrecan; axon regeneration; base; blood-brain barrier crossing; chemical standard; clinically relevant; design; druggable target; economic cost; effective therapy; in vitro Assay; in vivo; in vivo Model; in vivo regeneration; innovation; insight; neuroprotection; protein structure; repaired; screening; small molecule; small molecule libraries; small molecule therapeutics; success; transcriptome sequencing

A. ABSTRACT A significant impediment towards improvement of outcome after spinal cord injury (SCI) is the limited axonal regeneration due to insufficient regrowth of the axonal fibers at the lesion site. Signaling mediated by RhoA GTPases presents a significant impediment to growth. To date, there exists no small-molecule RhoA antagonists that have shown in vivo efficacy. This is attributed to the fact that small GTPases like RhoA do not have druggable pockets, making the development of small molecules that directly bind to them extremely difficult. However, over the years, several proteins with druggable pockets have been discovered to interact with RhoA. We stipulated that small molecules that bind to these druggable targets will promote axonal regeneration and recovery to a similar or greater extent than direct inhibition of RhoA activity. To identify such compounds we pursued a unique approach that consisted of structure-based computational docking of large chemical libraries to druggable targets that interact with RhoA. We tested the top 50-100 compounds for their effect on neurite outgrowth and axonal regeneration. One compound, RLA-47, inhibited in vitro neurite outgrowth and axonal regeneration. The compound also showed substantial axon crossings in an animal model of spinal cord injury. Our central hypothesis is that RLA-47 and its derivatives will promote axonal regeneration and recovery following spinal cord injury. Our preliminary data puts us in a strong position to test our hypothesis. In our first aim, we propose to design and synthesize derivatives of RLA-47 that will be tested in our neurite outgrowth and axonal regeneration in vitro assays. The most promising candidates will be explored in our in vivo hemisection model of spinal cord injury. In our second aim, we propose to develop RLA-47 derivatives with suitable pharmacokinetics and blood- brain permeability that promote axonal regeneration in vivo. We also test the most promising compounds using a more clinically relevant contusion model of spinal cord injury. We will use a standard chemical biology strategy of affinity purification and mass spectrometry to uncover targets of RLA-47 and its derivatives. Along with RNA sequencing, these studies will provide deeper insight into the mechanism of action of compounds. We expect 1-2 derivatives of RLA-47 to exhibit superior pharmacokinetics, brain permeability and efficacy. These compounds will lead to investigational new drug filing that we expect will eventually lead to candidates that can be explored in clinical trials either alone, or in combination with other therapies to promote regeneration and recovery in spinal cord injury patients.

A.摘要 有限的轴突有限 由于病变部位的轴突纤维再生不足而导致的再生。由Rhoa介导的信号传导 GTPASE对生长产生了重大障碍。迄今为止，没有小分子rhoa拮抗剂 在体内表现出的。这归因于以下事实：像Rhoa这样的小gtpase无法吸毒 口袋，使直接与它们结合的小分子的发展极为困难。但是，结束了 多年来，已经发现了几种带有可药袋的蛋白质与Rhoa相互作用。我们规定了 与这些可毒靶标结合的小分子将促进轴突再生并恢复到 与直接抑制RhoA活性相似或更大的程度。为了识别这样的化合物，我们追求了独特的 大型化学库的基于结构的计算对接的方法 与Rhoa相互作用。我们测试了前50-100种化合物对神经突生长和轴突的影响 再生。一种化合物RLA-47抑制了体外神经突生长和轴突再生。这 化合物在脊髓损伤的动物模型中还显示出大量的轴突横梁。我们的中心 假设是RLA-47及其衍生物将促进脊髓后轴突再生和恢复 受伤。我们的初步数据使我们处于检验假设的强烈立场。在我们的第一个目标中，我们建议 RLA-47的设计和合成衍生物将在我们的神经突生长和轴突再生中进行测试 体外测定。最有希望的候选者将在我们的体内脊髓半镜模型中探讨 受伤。在我们的第二个目标中，我们建议开发具有合适药代动力学和血液的RLA-47衍生物 促进体内轴突再生的脑渗透性。我们还使用 脊髓损伤的更临床相关的挫伤模型。我们将使用标准化学生物学策略 与发现RLA-47及其衍生物靶标的亲和力纯化和质谱。与RNA一起 测序，这些研究将为化合物的作用机理提供更深入的了解。我们期望 RLA-47的1-2衍生物表现出优异的药代动力学，脑部渗透性和功效。这些 化合物将导致我们期望的调查新药物提交，最终将导致候选人可以 可以在临床试验中进行探讨，或与其他疗法结合使用以促进再生和 脊髓损伤患者的恢复。