Testing OKN-007 as a potential intervention for ALS

测试 OKN-007 作为 ALS 的潜在干预措施

• 批准号: 10513312
• 负责人: HOLLY VAN REMMEN
• 金额: --
• 依托单位: OKLAHOMA CITY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-10-01 至 2023-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10513312
• 关键词: ALS patients; Address; Affect; Age; Aging; Amyotrophic Lateral Sclerosis; Anti-Inflammatory Agents; Antioxidants; Astrocytes; Atrophic; Brain; Cell Death; Clinic; Clinical Trials; Cochlea; Data; Denervation; Disease; Disease Progression; Distal; Dose; Effectiveness; Event; Gastrocnemius Muscle; Gene Expression; Generations; Glioblastoma; Goals; Hair Cells; Hand Strength; Health; Hindlimb; Human; Intervention; Ischemia; Laboratories; Maintenance; Malignant Neoplasms; Measures; Mediating; Microglia; Morphology; Motor Neurons; Mus; Muscle; Muscle function; Muscular Atrophy; Neuromuscular Diseases; Neuromuscular Junction; Neurons; Onset of illness; Outcome; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phase II Clinical Trials; Phenotype; Pilot Projects; Preventive; Preventive measure; Property; Reactive Oxygen Species; Skeletal Muscle; Spin Trapping; Spinal Cord; Stroke; Testing; Therapeutic Effect; Traumatic Brain Injury; Work; age related; amyotrophic lateral sclerosis therapy; axonopathy; cohort; drinking water; effective therapy; effectiveness testing; end stage disease; hearing impairment; improved; in vivo; inflammatory marker; interest; military veteran; mitochondrial dysfunction; mouse model; muscle form; mutant mouse model; nerve supply; neuromuscular; neuron loss; neuroprotection; nitrone; oxidative damage; phenyl-N-tert-butylnitrone; preservation; response; sarcopenia; small molecule; transcriptome sequencing; treatment effect

Amyotrophic lateral sclerosis (ALS) is a debilitating disease with currently no effective treatments. The goal of this proposal is to define the potential effectiveness of and mechanism of action of a promising new compound, OKN-007, that we have recently found to have neuroprotective effects in motor neurons in aging and in preliminary studies in an ALS mutant mouse model. Our recent studies in aging and sarcopenia have led us to focus on interventions targeted to preserving motor neuron health to reduce loss of innervation and NMJ disruption and therefore improve muscle outcomes in aging. To this end, we have exciting preliminary data using OKN-007, small molecule that reduces loss of motor neurons in aging mice. OKN-007 is a nitrone derivative of PBN (α-phenyl-N-tert-butyl-nitrone), a spin trap that has previously been shown to have beneficial effects in cancer and other pathologies. Nitrone compounds have known antioxidant and anti-inflammatory properties, and OKN-007 has neuroprotective effects in traumatic brain injury, in focal ischemia/stroke and in hearing loss. Our exciting preliminary data in aging mice show that OKN-007 can reduce motor neuron loss, reduce denervation in hindlimb muscle and reduce loss of gastrocnemius muscle mass in older mice. These findings led us to hypothesize that OKN-007 may also reduce motor neuron death and delay disease progression in ALS. Indeed, we are encouraged by our preliminary data that show a preservation of motor neuron number in the spinal cord and a delay in disease progression in the G93A ALS mouse model treated with OKN-007. The goal of this pilot project is to confirm these preliminary findings and test our hypothesis that OKN-007 can preserve motor neuron loss in ALS and delay disease progression. In Aim 1, we will determine the effect of OKN-007 treatment on motor neuron and skeletal muscle outcomes in disease progression and survival in the G93A mutant mouse model of ALS. Cohorts of control and G93A mice will be treated with OKN-007 in drinking water beginning at 30 days of age, at disease onset (95 days), or at 3 weeks post-onset (115 days) to determine preventative and therapeutic effects of the treatment. Mice will weighed and evaluated for disease score and progression beginning at 60 days of age. Using spinal cord from control and G93A mice, we will measure motor neuron number, inflammatory markers, oxidative damage and activation of cell death, known alterations in disease progression in ALS. Mice will be sacrificed at 145 days of age, prior to disease end stage. To assess known ALS mediated muscle phenotypes, we will measure grip strength starting 60 days of age and NMJ morphology and innervation status will be measured at 145 days, as well as markers of denervation in skeletal muscle and muscle atrophy. Parallel cohorts of mice will be set up to measure survival in response to drug treatment once we determine the most effective dose. In Aim 2, we will identify potential mechanisms of action for OKN-007 in motor neuron maintenance and survival by measuring the effect of OKN-007 on gene expression using RNA seq analysis in spinal cord motor neurons from mice in Aim 1. These studies will provide important information on the mechanism of action of OKN-007 and help to establish whether OKN-007 may be a good candidate for treatment to maintain motor neuron health in ALS patients.

肌萎缩侧索硬化症（ALS）是一种使人衰弱的疾病，目前尚无有效的治疗方法。目标是 该提案旨在定义一种有前途的新化合物的潜在有效性和作用机制， OKN-007，我们最近发现它对衰老和衰老过程中的运动神经元具有神经保护作用 ALS 突变小鼠模型的初步研究。我们最近对衰老和肌肉减少症的研究使我们发现 重点关注旨在保护运动神经元健康的干预措施，以减少神经支配和 NMJ 的丧失 破坏，从而改善衰老过程中的肌肉结果。为此，我们有令人兴奋的初步数据 使用 OKN-007，一种可以减少衰老小鼠运动神经元损失的小分子。 OKN-007是一种硝酮 PBN（α-苯基-N-叔丁基硝酮）的衍生物，一种自旋陷阱，之前已被证明具有有益的作用 对癌症和其他病理的影响。硝酮化合物具有抗氧化和抗炎作用 OKN-007 对创伤性脑损伤、局灶性缺血/中风和 听力损失。我们在衰老小鼠中的令人兴奋的初步数据表明 OKN-007 可以减少运动神经元损失， 减少老年小鼠后肢肌肉的去神经支配并减少腓肠肌质量的损失。这些 研究结果使我们推测 OKN-007 也可能减少运动神经元死亡并延缓疾病 ALS 的进展。事实上，我们对我们的初步数据感到鼓舞，这些数据显示了运动能力的保存 G93A ALS 小鼠模型中脊髓神经元数量和疾病进展延迟 用 OKN-007 处理。该试点项目的目标是确认这些初步发现并测试我们的 假设 OKN-007 可以保留 ALS 中运动神经元的损失并延缓疾病进展。在 目标 1，我们将确定 OKN-007 治疗对运动神经元和骨骼肌结果的影响 ALS G93A 突变小鼠模型的疾病进展和生存。对照组小鼠和 G93A 小鼠组 将在 30 天龄、疾病发作时（95 天）或 3 岁开始在饮用水中接受 OKN-007 处理。 发病后几周（115 天）以确定治疗的预防和治疗效果。老鼠会 从 60 日龄开始称重并评估疾病评分和进展情况。使用脊髓 对照小鼠和 G93A 小鼠，我们将测量运动神经元数量、炎症标记物、氧化损伤和 细胞死亡的激活，ALS 疾病进展的已知改变。小鼠将在145天处死 疾病末期之前的年龄。为了评估已知的 ALS 介导的肌肉表型，我们将测量握力 力量从 60 天开始测量，NMJ 形态和神经支配状态将在 145 天测量，如下 以及骨骼肌去神经支配和肌肉萎缩的标志物。将建立平行的小鼠队列 一旦我们确定了最有效的剂量，就可以衡量对药物治疗的生存率。在目标 2 中，我们将 通过测量确定 OKN-007 在运动神经元维持和存活中的潜在作用机制 使用 RNA seq 分析确定 OKN-007 对小鼠脊髓运动神经元基因表达的影响 目标 1. 这些研究将提供有关 OKN-007 作用机制的重要信息，并有助于 确定 OKN-007 是否是维持 ALS 运动神经元健康的良好治疗候选者 患者。