Small-Molecule Pharmacological Chaperones to Prevent Synuclein Transmission

防止突触核蛋白传播的小分子药理学伴侣

• 批准号: 9114683
• 负责人: LISA C MCCONLOGUE
• 金额: $ 47.33万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9114683
• 关键词: Affect; Agreement; Alzheimer' s Disease; Animal Disease Models; Animal Model; Binding; Biochemical; Biological; Biological Assay; Brain; Cell model; Cells; Cellular Assay; Characteristics; Collaborations; Data; Development; Diffuse; Diffuse Lewy Body Disease; Disease; Disease Progression; Educational process of instructing; Event; Fluorescence Resonance Energy Transfer; Foundations; Functional disorder; Future; Goals; Grant; Health; Human; Legal; Lewy Bodies; Light; Location; Measures; Mediating; Microscopic; Modeling; Molecular Chaperones; Molecular Conformation; Morphologic artifacts; Nerve Degeneration; Neurodegenerative Disorders; Parkinson Disease; Pathology; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Process; Proteins; Research; Research Project Grants; Resources; Staging; Structure-Activity Relationship; Techniques; Testing; Therapeutic; Toxic effect; Universities; aggregation pathway; alpha synuclein; analog; base; extracellular; in vivo; in vivo Model; innovation; novel; novel strategies; novel therapeutics; overexpression; prevent; prion-like; screening; small molecule; synuclein; synucleinopathy; therapeutic development; trafficking; translational study; transmission process; uptake

 DESCRIPTION (provided by applicant): Our overall goal is to discover small molecule chaperones targeting -synuclein (Syn) that prevent the prion-like transmission of misfolded Syn and can be developed into drugs to treat neurodegenerative diseases such as Parkinson's (PD), Diffuse Lewy Body, and Alzheimer's diseases. These drugs will act via a novel mechanism of action by directly interacting with Syn to prevent spreading of Syn in the brain, with the ultimate goal of reducing disease progression. We envision an innovative strategy by which small molecules bind to and stabilize native or other non-pathogenic states of Syn and thereby block pathological events. Most small molecules affecting Syn aggregation were found in aggregation assays and interact with aggregation pathway forms of Syn. By contrast, we will test compounds that bind to Syn native states and affect Syn dysfunction more broadly. From a biophysical screen we previously identified 61 compounds that bind to native Syn (Syn-PCs) some of which prevented Syn-mediated dysfunctions such as Syn aggregation and Syn-induced vesicular dysfunction. Based on these promising results, we propose to expand the development of Syn-PCs as novel Syn chaperone drugs. In this application, we will determine if Syn-PCs are a viable strategy to limit Syn cell-to-cell transmission and explore the mechanism of action of such active compounds. We will first identify Syn-PCs capable of preventing the cell-to-cell transmission of Syn and in so doing, identify molecules having potential to be developed into therapeutic drugs. In Aim 1 we will screen the Syn-PCs in cellular assays of Syn transmission including a FRET-based assay in which transmission is measured as a direct association of donor Syn with acceptor Syn expressed in different cells. Based on preliminary data we expect to find active compounds. Structure activity relationships will be explored by testing activity of commercially available analogues of active compounds. In Aim 2 we will further identify which steps of transmission are modulated by different Syn-PCs. Different compounds will likely affect Syn transmission by different mechanisms (e.g., altered cellular levels, location, expulsion, uptake, misfolding) and thus novel mechanisms are likely to be identified using Syn-PCs. It will be greatly informative for further therapeutic optimization to explore these novel mechanisms. We will analyze the active Syn-PCs to identify which stage in transmission is blocked. Biochemical, microscopic and cell biological techniques will be applied to explore the cellular and extracellular location and state of Syn in media and donor and recipient cells. These analyses will elucidate how active Syn-PCs modulate the transmission of Syn and will shed light on possible assays that would be useful for future development of these molecules into drugs to treat patients. These studies will provide the first evidence for efficacy of Syn chaperones in blocking the transmission of a critical protein involved in PD progression and pathology and will provide the foundation for the development of a novel class of drugs for treating Syn related neurodegenerative diseases.

 描述（由申请人提供）：我们的总体目标是发现针对α-突触核蛋白（αSyn）的小分子伴侣，可以防止错误折叠的αSyn的类似朊病毒的传播，并可以开发成治疗神经退行性疾病的药物，例如帕金森病（PD）、弥漫性路易体病和阿尔茨海默病。这些药物将通过一种新颖的作用机制发挥作用，直接与 αSyn 相互作用，以防止 αSyn 在体内扩散。 大脑，最终目标是减少疾病进展。我们设想了一种创新策略，通过该策略，小分子结合并稳定 αSyn 的天然或其他非致病状态，从而阻止病理事件。大多数影响 αSyn 聚集的小分子是在聚集测定中发现的，并与 αSyn 的聚集途径形式相互作用。相比之下，我们将测试与 αSyn 天然状态结合并更广泛地影响 αSyn 功能障碍的化合物。通过生物物理筛选，我们之前鉴定了 61 种与天然 αSyn (αSyn-PC) 结合的化合物，其中一些可以预防 αSyn 介导的功能障碍，例如 αSyn 聚集和 αSyn 诱导的囊泡功能障碍。基于这些有希望的结果，我们建议扩大 αSyn-PC 作为新型 αSyn 伴侣药物的开发。在此应用中，我们将确定 αSyn-PC 是否是限制 αSyn 细胞间传播的可行策略，并探索此类活性化合物的作用机制。 我们将首先鉴定能够阻止 αSyn 细胞间传播的 αSyn-PC，并在此过程中鉴定有潜力开发成治疗药物的分子。在目标 1 中，我们将在 αSyn 传递的细胞测定中筛选 αSyn-PC，包括基于 FRET 的测定，其中传递被测量为供体 αSyn 与在不同细胞中表达的受体 αSyn 的直接关联。根据初步数据，我们期望找到活性化合物。将通过测试活性化合物的市售类似物的活性来探索结构活性关系。在目标 2 中，我们将进一步确定哪些传输步骤是由不同的 Syn-PC 调制的。不同的化合物可能会通过不同的机制（例如，改变细胞水平、位置、排出、摄取、错误折叠）影响 αSyn 传递，因此可能使用 αSyn-PC 来识别新的机制。探索这些新机制将为进一步治疗优化提供大量信息。我们将分析活动的 Syn-PC，以确定传输中的哪个阶段被阻止。将应用生化、显微镜和细胞生物学技术来探索 Syn 在培养基以及供体和受体细胞中的细胞和细胞外位置和状态。这些分析将阐明活性 αSyn-PC 如何调节 αSyn 的传递，并将揭示可能的分析方法，这些分析方法将有助于未来将这些分子开发成治疗患者的药物。 这些研究将为αSyn伴侣在阻断PD进展和病理学中涉及的关键蛋白质的传递方面的功效提供第一个证据，并将为开发一类用于治疗αSyn相关神经退行性疾病的新型药物奠定基础。