Development of a Synthetic, Cell-Permeable Peptide for the Inhibition of CDK5-p25 Hyperactivity in Amyotrophic Lateral Sclerosis

项目摘要-在这项直接到第二阶段的SBIR应用中，Cogentis Treateutics计划推进 治疗致病CDK5-p25的合成细胞通透性多肽抑制剂CT526的研究进展 用于肌萎缩侧索硬化症(ALS)的神经退行性变。Cogentis已经证明，CT526使 CDK5在ALS小鼠模型中的病理活性，同时保留了基本的CDK5功能，但 还需要更多的工作来表征其在表型多样化和临床相关的小鼠中的表现 人类IPSC模型。此外，还需要配套的生物标志物来跟踪对CT526的早期反应 临床前/临床研究，并最终支持临床决策，包括药物滴定。考虑到 FDA批准的两种治疗肌萎缩侧索硬化症的药物只提供了适度的益处，进一步的开发和商业化 CT526及其伴生生物标记物的研究可能大大推进ALS的药物治疗，潜在地提供 延缓、阻止或逆转神经退行性变及其对病人质量的破坏性影响的第一种疗法 生命和生存的关系。目的1.在表型多样的ALS小鼠模型中证明CT526的有效性。 里程碑：在治疗组中，1)显著延迟了疾病的发病，延长了寿命，和/或 改善步态和后凸评分进展；2)显示CDK5过度活动表型正常化 P25代在野生型对照水平的10%以内；3)P-TDP43和NF-L在 脑脊液；4)脊髓P-TDP43和炎性标志物减少。(P&lt；0.05 对于所有差异。)目的2.证实CT526对人IPSCs的保护作用 ALS最常见的分子亚型。里程碑：1)验证IPSC细胞系中CDK5的表型 与正常对照组相比，肌萎缩侧索硬化症患者≥的CDK5活性增加20%或≥的p25/p35比值增加20%或更多。 健康供体IPSCs；2)在谷氨酸毒性检测中显示IPSCs的剂量反应保护作用 CT526治疗。目的3.表征基于神经衍生外切体(NDE)的生物标志物的反应 去CT526。里程碑：1)显示≥在任何亚组的CDK5活性读数都增加了20% ALS的死后脑与非ALS年龄匹配的对照组进行比较。2)对肌萎缩侧索硬化症患者进行亚型分类 CDK5的过度活跃和生物标志物的变化程度。3)相关生物标志物来自死后脑和 RHO&GT；0.65，疾病亚组与对照亚组≥相差20%，AUC≥相差80%。 4)检测CT526治疗后≥高活性相关NDE生物标志物中CDK520%的减少 AIMS 1和AIMS 2中的鼠标/IPSC模型。Impact-该项目预计将在 表型多样化和临床相关的模型，并确定可使用的基于无损检测的生物标记物(S) 跟踪ALS治疗的临床前和临床研究中的药物反应性。在未来的工作中，Cogentis 将a)利用生物标记物(S)作为优化CT526给药策略的工具，b)实施IND CT526的安全性、有效性和毒性研究。

PROJECT SUMMARY—In this Direct-to-Phase II SBIR application, Cogentis Therapeutics proposes to advance the development of CT526, a synthetic, cell-permeable peptide inhibitor of pathogenic CDK5-p25 as a treatment for neurodegeneration in amyotrophic lateral sclerosis (ALS). Cogentis has shown that CT526 normalizes the pathological activity of CDK5 in a mouse model of ALS while preserving essential CDK5 functionality, but additional work is needed to characterize its performance in phenotypically diverse mouse and clinically relevant human iPSC models. In addition, companion biomarkers are needed to track early responsiveness to CT526 in preclinical / clinical studies and to eventually support clinical decision-making, including drug titration. Given that the two FDA-approved drugs for ALS only provide modest benefit, further development and commercialization of CT526 and its companion biomarkers may substantially advance drug therapy for ALS, potentially providing the first treatment that slows, stops, or reverses neurodegeneration and its devastating effects on patient quality of life and survival. Aim 1. Demonstrate CT526 efficacy in a phenotypically diverse mouse model of ALS. Milestones: In the treatment group, 1) Demonstrate significant delay in disease onset, prolonged lifespan, and/or improved gait and kyphosis score progression; 2) Demonstrate normalization of CDK5 hyperactivity phenotype and p25 generation within 10% of the wildtype control level; 3) Demonstrate reduction of P-TDP43 and NF-L in cerebrospinal fluid; and 4) Demonstrate reduction of P-TDP43 and inflammatory markers in spinal cord. (P<0.05 for all differences.) Aim 2. Demonstrate the protective effect of CT526 in human iPSCs from patients with the most common molecular subtype of ALS. Milestones: 1) Validate CDK5 phenotype in iPSC cell lines by demonstrating ≥ 20% increase in CDK5 hyperactivity or ≥ 20% or more increase in p25/p35 ratio in ALS vs. healthy donor iPSCs; 2) Demonstrate dose-responsive protection of iPSCs in glutamate toxicity assays when treated with CT526. Aim 3. Characterize the response of neural-derived exosome (NDE)-based biomarkers to CT526. Milestones: 1) Demonstrate ≥ 20% increase in any of the CDK5 activity readouts in any subgroup of ALS postmortem brains compared to non-ALS age-matched controls. 2) Categorize ALS patient subtypes by CDK5 hyperactivity and degree of change in biomarkers. 3) Correlate biomarkers from postmortem brain and from NDE analysis with rho > 0.65, ≥ 20% difference between disease and control subgroups, and AUC ≥ 80%. 4) Detect ≥ 20% reduction in CDK5 hyperactivity-related NDE biomarkers in response to CT526 treatment of mouse/iPSC models in Aims 1 and 2. Impact—This project is expected to confirm CT526 efficacy in phenotypically diverse and clinically relevant models and identify an NDE-based biomarker(s) that can be used to track drug responsiveness in preclinical and clinical studies for an ALS therapeutic. In future work, Cogentis will a) leverage the biomarker(s) as a tool for optimizing CT526 dosing strategies and b) conduct IND-enabling safety, efficacy, and toxicity studies of CT526.