In vivo Evaluation of USP30 Inhibitors in Models Relevant to Parkinson's Disease

摘要 影响全球1000万人的帕金森病(PD)的现有治疗方法主要是增强 多巴胺能神经传递对症状有好处。目前没有一种治疗方法可以延缓或阻止 帕金森病的进展。我们提出了一个药物发现平台来开发针对Parkin-USP30的小分子 有丝分裂途径，它代表了线粒体动态平衡的关键调节因素。会聚的线条 来自散发性帕金森病患者和动物的人类遗传学、组织病理学和生化研究的证据 模型研究表明，吞丝分裂的缺陷是帕金森病发病的一个可修改的因素。具体来说， 抑制或敲除去泛素化酶USP30可以增强多种细胞的有丝分裂能力 并推断了两种果蝇模型(PINK1缺乏和百草枯中毒)的神经保护作用。然而，研究表明 研究啮齿动物等大型动物的神经退行性变是知识上的一个空白，本应 在开发USP30抑制剂作为帕金森病的治疗药物之前。在提议的目标中，我们将测试 假设USP30抑制剂小分子将保护由A诱导的神经变性 线粒体毒素MPTP，或a-突触核蛋白预制纤维-这两种模型都与帕金森病和 有丝分裂。我们的初步数据表明，我们已经产生了一系列有效的小分子 并在体外选择性地抑制USP30的活性，并在分化的细胞中表现出无细胞毒性的细胞活性 神经细胞系。TOP化合物具有良好的体外ADME性能，而先导和后备化合物具有 通过出色的大鼠和小鼠血浆和脑PK图谱进行鉴定，这使我们处于独特的位置 检验提出的假设。如果成功，本申请中提出的研究将进一步证明 通过启用研究性新药备案(IND)投资于我们的小分子开发 研究，获得许可，或与大型制药公司或生物技术公司合作进行进一步的临床开发。

ABSTRACT Existing treatments for Parkinson’s disease (PD), which affects 10 million people worldwide, primarily augment dopaminergic neurotransmission to provide symptomatic benefit. No current therapy can slow or stop the progression of PD. We propose a drug discovery platform to develop small molecules targeting the parkin-USP30 mitophagy pathway, which represents a key regulator of mitochondrial homeostasis. Converging lines of evidence from human genetics, tissue pathology and biochemical studies from sporadic PD patients, and animal model studies indicate that deficits in Mitophagy are a modifiable contributor to PD pathogenesis. Specifically, inhibition or knockdown of the deubiquitinating enzyme USP30 can enhance mitophagy in a variety of cell types and infers neuroprotection in two fruit fly models (pink1 deficiency and paraquat toxicity). However, studies examining neurodegeneration in larger animals such as rodents are an unfilled gap in knowledge that should be addressed prior to development of USP30 inhibitors as therapeutics for PD. In the proposed Aims, we will test the hypothesis that USP30 inhibitor small molecules will protect against neurodegeneration induced by a mitochondrial toxin MPTP, or a-synuclein preformed fibrils – both models with links to Parkinson’s disease and mitophagy. Our preliminary data demonstrate that we have generated a series of small molecules that potently and selectively inhibit USP30 activity in vitro, and demonstrate cellular activity without cytotoxicity in differentiated neuronal cell lines. Top compounds have good in vitro ADME properties and a lead and back up compound have been identified with excellent rat and mouse plasma and brain PK profiles which place us in a unique position to examine the proposed hypothesis. If successful, the studies proposed in this application will justify further investment toward development of our small molecules through Investigational New Drug filing (IND)-enabling studies, out-license, or partnership with big pharma or biotech for further clinical development.