Pharmacokinetic and pharmacodynamic assessment of peptide-based therapy DA1 for the treatment of Alzheimer's disease

项目摘要/摘要 JanusQ，LLC是一家初创生物技术公司，从凯斯西储大学剥离出来，正在开发 一种基于多肽的疗法治疗阿尔茨海默病(AD)中的线粒体功能障碍，这是 侵蚀了大约600万美国人的记忆和认知能力。淀粉样蛋白β(Aβ)在阿尔茨海默病中的作用 发病机制尚不清楚，针对A的治疗在临床上并不成功。这个 观察到，像许多其他神经退行性疾病一样，AD与线粒体功能障碍有关， 导致神经毒性提示了一种新的策略：ATAD3A是一种线粒体蛋白质，它跨越内部和 外膜是胆固醇运输和维持线粒体类核复合体的关键。 然而，在AD应激过程中，ATAD3A寡聚并与Drp1(线粒体裂变GTP酶)结合， 导致线粒体碎裂和类核不稳定。DA1，一种由JanusQ新发现的肽 共同创建者，防止ATAD3A寡聚和与Drp1的结合，减少线粒体碎裂， 改善类核的稳定性，并减少AD和AD啮齿动物模型的行为和神经缺陷 亨廷顿氏病。然而，对DA1在体内的分布和剂量依赖关系知之甚少。因此， 很难知道未经修饰的DA1是否已经适合作为治疗AD的临床候选药物， 或者是否需要进一步的工作来优化多肽以改善其药理特性 (效力、吸收、分配、消除)。这笔SBIR赠款的目的就是做出这一决定。 该项目有三个目标：(1)评估DA1在体内的药代动力学，以测试暴露和稳定性，通过 开发从血浆和脑组织中提取DA1的方法，在不同时间点采集样本 在皮下注射多肽(SQ)，并使用串联液体定量多肽 色质联用。(2)进行DA1的初步体外安全性/毒性评价，重点 关于发现阶段的分析，评估潜在的脱靶相互作用和免疫原性。(3) 使用体内AD小鼠模型评估DA1剂量-反应特性，治疗APP敲入AD小鼠 (3-9个月大)，最初服用三剂DA1。验证分析将评估ATAD3A寡聚， 线粒体生物能量学，AD病理测量(例如，Aβ积聚、突触完整性、神经元丢失)， 以及AD发展过程中的认知功能。这些结果将直接解决我们的核心假设，即 DA1具有进入临床所必需的药理特性(与未来的多肽相比 优化)。因此，拟议的工作是朝着开发多肽的长期目标迈出的合乎逻辑的下一步 改善AD患者的生活质量和生存的治疗。这样的治疗将减少对医疗的依赖 护理人员，从而降低总体医疗成本。在美国，仅直接医疗成本在2020年就超过了3050亿美元。 这笔SBIR拨款是JanusQ开发DA1本身(或首先优化DA1)作为突破的关键一步 治疗阿尔茨海默病。无论哪种方式，都会让JanusQ处于吸引商业化合作伙伴的地位。

PROJECT SUMMARY/ABSTRACT JanusQ, LLC is a startup biotech company, spun out of Case Western Reserve University, and developing a peptide-based therapy for treatment of mitochondrial dysfunction in Alzheimer’s Disease (AD), which is eroding the memory and cognitive abilities of ~6 million Americans. The role of amyloid-β (Aβ) in AD pathogenesis is poorly understood, and therapies addressing A have not been successful in the clinic. The observation that, like many other neurodegenerative diseases, AD is linked to mitochondrial dysfunction that leads to neurotoxicity suggests a novel strategy: ATAD3A is a mitochondrial protein that spans both inner and outer membranes and is critical in cholesterol trafficking and maintaining the mitochondrial nucleoid complex. However, during the stress of AD, ATAD3A oligomerizes and binds Drp1 (the mitochondrial-fission GTPase), causing mitochondrial fragmentation and nucleoid instability. DA1, a peptide newly discovered by a JanusQ cofounder, prevents ATAD3A oligomerization and association with Drp1, reduces mitochondrial fragmentation, improves nucleoid stability, and reduces behavioral and neurological deficits in rodent models of AD and Huntington’s disease. However, little is known about the disposition and dose-dependence of DA1 in vivo. Thus, it is difficult to know whether unmodified DA1 is already suitable as a clinical candidate for the treatment of AD, or whether future work will be required to optimize the peptide to improve its pharmacological characteristics (potency, absorption, distribution, elimination). The purpose of this SBIR grant is to make that determination. The project has three aims: (1) Assess the pharmacokinetics of DA1 in vivo to test exposure and stability by developing approaches to extract DA1 from plasma and brain tissue, collecting samples at various time points after administering the peptide subcutaneously (SQ), and quantitating the peptide using tandem liquid chromatography mass spectroscopy. (2) Perform preliminary in vitro safety/toxicity assessment of DA1, focusing on discovery-phase assays, with an assessment of potential off-target interactions and immunogenicity. (3) Assess DA1 dose-response characteristics using an in vivo AD mouse model, treating APP knock-in AD mice (aged 3–9 months) initially with three DA1 doses SQ. Validation assays will assess ATAD3A oligomerization, mitochondrial bioenergetics, measures of AD pathology(e.g., Aβ accumulation, synaptic integrity, neuronal loss), and cognitive function during AD development. These results will directly address our central hypothesis that DA1 has the pharmacological properties necessary for advancement to the clinic (vs. future peptide optimization). Thus, the proposed work is the next logical step toward the long-term goal of developing a peptide therapy to improve quality of life and survival of AD patients. Such a treatment would reduce reliance on medical caregivers and thus reduce overall medical costs. In the US, direct medical costs alone exceeded $305B in 2020. This SBIR grant is a crucial step for JanusQ to develop DA1 per se (or first to optimize DA1) as a breakthrough treatment for AD. Either approach would put JanusQ in a position to attract a commercialization partner.