New Drug Discovery Paradigms for Synucleinopathies

突触核蛋白病的新药物发现范例

• 批准号: 9392291
• 负责人: DAVID R BORCHELT
• 金额: $ 25.87万
• 依托单位: LANKENAU INSTITUTE FOR MEDICAL RESEARCH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9392291
• 关键词: Adopted; Algorithms; Alzheimer' s Disease; Amino Acids; Amyloid beta-Protein; Area; Autopsy; Binding; Biological Assay; Brain; Buffers; Chemicals; Clinic; Complement; Cytology; Diagnosis; Dimerization; Disease; Drug Combinations; Exhibits; FDA approved; Functional disorder; Genetic; Individual; Investigation; Lead; Lewy Bodies; Lewy Body Dementia; Ligand Binding; Lipids; Luciferases; Membrane; Methods; Molecular Conformation; Molecular Weight; Neurodegenerative Disorders; Neurofibrillary Tangles; Neurosciences Research; Outcomes Research; Parkinson Disease; Parkinson' s Dementia; Pathogenicity; Pathology; Pathway interactions; Peptides; Pharmaceutical Preparations; Phospholipids; Preclinical Drug Development; Proteins; Research; Senility; Specificity; Structure; Surface; Tauopathies; Testing; Therapeutic; Translating; Vascular Dementia; Vesicle; Vitronectin; alpha synuclein; combinatorial; conformer; data mining; dimer; drug candidate; drug discovery; gain of function; high throughput screening; inhibitor/antagonist; innovation; misfolded protein; monomer; novel; novel strategies; novel therapeutics; patient population; premature; prevent; protein aggregate; protein aggregation; protein misfolding; protein phosphatase inhibitor-2; protein protein interaction; proteostasis; reconstitution; research clinical testing; screening; small molecule; synergism; synucleinopathy; tau Proteins; translational neuroscience

Summary: New Drug Discovery Paradigms for Synucleinopathies Dysfunction in cellular proteostasis leads to abnormal accumulation of misfolded proteins implicated in the pathology of several neurodegenerative diseases. Alpha-synuclein (aS) is the primary component of intracellular inclusions known as Lewy bodies—the cytopathological hallmark of Lewy body dementias, Parkinson’s disease and a frequent pathology in Alzheimer’s disease (AD). Both aS and tau pathology may overlap in AD because of the potential for inter-nucleation and aggregation of these two proteins. Thus, synucleinopathy is a high-priority target that bridges several CNS protein-misfolding disorders. Alpha-synuclein is a 140-amino-acid intracellular protein. It is an intrinsically disordered monomer but can adopt multiple - helical conformations on binding to lipid vesicles. This transition from disordered to -helical conformations is thought to lead to the formation of misfolded -sheet-rich structures that can form soluble oligomers and aggregates. Our hypothesis is that aS dimerization induced by membrane phospholipids is an early, rate- limiting step in protein-misfolding pathways that ultimately leads to synucleinopathies; and that a drug inhibiting this step will have therapeutic value. We have developed an ex vivo split-luciferase protein complementation assay to detect the initial dimerization of aS as it oligomerizes and aggregates. Our proposal is to use this assay in two innovative, high throughput screening (HTS) paradigms to identify novel aS aggregation inhibitors. One approach will take the standard long road to identify novel small molecules. We will screen 100,000 compounds in assay buffer conditioned with phospholipids that enhance aS dimerization. A panel of secondary assays will be used to characterize confirmed HTS hits for specificity, potency, mode of action and cellular activity. The expected Aim 1 milestone is identification of a novel lead suitable for nomination as a preclinical drug development candidate. Because of the mismatch between the small sizes of drug-like molecules compared to the expansive surface area involved in protein-protein interactions (PPI), HTS for protein-misfolding inhibitors is challenging. To potentially accelerate getting new therapies to the clinic, we propose a more unconventional approach in which we will attempt to identify a combination of two FDA approved drugs acting supra-additively by allosteric synergy to prevent the self-association and aggregation of aS. We will use a multiplexed-HTS method we developed for screening mixtures of FDA-approved drugs to discover combinations that stabilize monomeric aS structure, preventing self-association. The expected Aim 2 milestone is identification of a combination of two previously approved drugs that can potentially advance to proof-of-concept clinical testing for treating synucleinopathies faster than an individual new chemical entity. If successful, the new approaches we develop to discover novel PPI modulators could have broader utility for advancing understanding of other neurodegenerative diseases caused by protein misfolding and aggregation.

摘要：突触核蛋白病的新药发现范式 细胞蛋白质稳态的功能障碍导致错误折叠的蛋白质的异常积累，这些蛋白质与细胞内的蛋白质代谢有关。 几种神经退行性疾病的病理学。α-突触核蛋白（aS）是 称为路易体的细胞内内含物-路易体痴呆的细胞病理学标志， 帕金森病和阿尔茨海默病（AD）中的常见病理。aS和tau病理学都可能 由于这两种蛋白质的成核和聚集的潜力，在AD中存在重叠。因此，在本发明中， 突触核蛋白病是连接几种CNS蛋白质错误折叠病症的高优先级靶标。阿尔法synuclein 是一种140个氨基酸的细胞内蛋白。它是一种本质上无序的单体，但可以采用多种结构， 螺旋构象对脂囊泡结合的影响。这种从无序到β-螺旋构象的转变是 被认为导致形成错误折叠的富含纸张的结构，该结构可以形成可溶性低聚物， 集料.我们的假设是，由膜磷脂诱导的aS二聚化是一种早期的，速率- 蛋白质错误折叠途径中的限制步骤，最终导致突触核蛋白病; 这一步骤将具有治疗价值。我们已经开发了一种离体分裂荧光素酶蛋白互补 测定以检测aS在寡聚和聚集时的初始二聚。我们的建议是利用这个 采用两种创新的高通量筛选（HTS）模式进行检测，以识别新型aS聚集 抑制剂的一种方法将采取标准的漫长道路来识别新的小分子。我们将筛选 在用磷脂调节的测定缓冲液中的100，000种化合物，其增强aS二聚化。一组 二次试验将用于表征已确认的HTS命中物的特异性、效价、作用模式和 细胞活动预期的目标1里程碑是确定适合提名为 临床前药物开发候选人。由于类药物的小尺寸之间的不匹配， 与蛋白质-蛋白质相互作用（PPI）中涉及的广阔表面积相比， 蛋白质错误折叠抑制剂具有挑战性。为了加速新疗法进入临床，我们 我提出了一个更非传统的方法，我们将尝试确定两个FDA的组合， 通过变构协同作用发挥超加和作用以防止自缔合和聚集的批准药物 如.我们将使用我们开发的用于筛选FDA批准药物混合物的多重HTS方法， 发现稳定单体aS结构的组合，防止自缔合。预期目标2 一个里程碑是确定了两种以前批准的药物的组合， 用于治疗突触核蛋白病的概念验证临床测试比单个新化学实体更快。如果 成功，我们开发的新方法，发现新的PPI调节剂可能有更广泛的用途， 推进对由蛋白质错误折叠和聚集引起的其他神经退行性疾病的理解。