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

项目总结-在此直接进入II期SBIR申请中，Cogentis Therapeutics建议推进 CT 526的开发，CT 526是一种致病性CDK 5-p25的合成的细胞渗透性肽抑制剂， 用于肌萎缩侧索硬化症（ALS）的神经变性。Cogentis已经表明，CT 526使 在ALS小鼠模型中的CDK 5的病理活性，同时保留基本的CDK 5功能，但 需要进一步的工作来表征其在表型多样的小鼠和临床相关的小鼠中的性能。 人类iPSC模型。此外，还需要伴随生物标志物来跟踪患者对CT 526的早期反应性。 临床前/临床研究，并最终支持临床决策，包括药物滴定。鉴于 FDA批准的两种用于ALS的药物仅提供适度的益处， CT 526及其伴随的生物标志物可能大大促进ALS的药物治疗， 第一种减缓、停止或逆转神经变性及其对患者质量的破坏性影响的治疗方法 生命和生存。目标1.证明CT 526在表型多样的ALS小鼠模型中的疗效。 糖尿病：在治疗组中，1）显示疾病发作显著延迟，寿命延长，和/或 改善步态和脊柱后凸评分进展; 2）证明CDK 5过度活跃表型正常化 和p25产生在野生型对照水平的10%内; 3）证明在野生型对照中P-TDP 43和NF-L的减少。 脑脊液;和4）证实脊髓中P-TDP 43和炎性标志物的减少。（P<0.05 所有的差异）。目标2.证明CT 526在来自患有前列腺癌的患者的人iPSC中的保护作用。 ALS最常见的分子亚型步骤：1）通过以下方法在iPSC细胞系中抑制CDK 5表型： 与对照组相比，ALS中CDK 5过度活跃增加≥ 20%或p25/p35比值增加≥ 20%或更多。 健康供体iPSC; 2）在谷氨酸毒性测定中证明iPSC的剂量响应性保护， 用CT 526治疗。目标3。表征基于神经源性外泌体（NDE）的生物标志物的反应 在CT 526。Mild：1）在任何亚组中，任何CDK 5活性读数显示增加≥ 20%。 ALS死后的大脑与非ALS年龄匹配的对照相比。2)ALS患者亚型分类 CDK 5活性亢进和生物标志物的变化程度。3)将死后大脑的生物标志物 来自NDE分析，rho > 0.65，疾病和对照亚组之间的差异≥ 20%，AUC ≥ 80%。 4)检测到CT 526治疗后CDK 5高活性相关NDE生物标志物降低≥ 20% 小鼠/iPSC模型中。影响-该项目预计将证实CT 526在以下方面的疗效： 表型多样性和临床相关模型，并确定可以使用的基于NDE的生物标志物 在临床前和临床研究中跟踪ALS治疗剂的药物反应性。在未来的工作中，Cogentis 将a）利用生物标志物作为优化CT 526给药策略的工具，和B）进行IND使能 CT 526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.