New Drug Discovery Paradigms for Synucleinopathies

突触核蛋白病的新药物发现范例

基本信息

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
项目类别：
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&apos 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&apos 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

项目摘要

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：新药物发现典范范例细胞蛋白抑制性功能障碍导致与该蛋白的异常积累几种神经退行性疾病的病理学。 α-核蛋白（AS）是细胞内夹杂物称为路易尸体 - 路易身体痴呆的细胞病理学标志，帕金森氏病和阿尔茨海默氏病（AD）的常见病理。 AS和Tau病理可能由于这两种蛋白质的核间和聚集的潜力，AD重叠。那，肌醇是一个高优先级的靶标，它桥接了几种CNS蛋白质不满意的疾病。 α-核蛋白是140-氨基酸的细胞内蛋白。它是一种本质上无序的单体，但可以采用多个- 与脂质蔬菜结合的螺旋构象。从无序到螺旋构象的过渡是被认为会导致形成错误折叠的表富含结构的结构，这些结构可以形成固体的低聚物和聚合。我们的假设是，正如膜磷脂诱导的二聚化是早期的，速率 - 限制蛋白质不满意途径的步骤，最终导致突触核酸的途径；并抑制药物此步骤将具有治疗价值。我们已经开发了一个离体分裂葡萄酸酶蛋白质的完成测定以检测AS的初始二聚化，以寡聚和聚集体。我们的建议是使用它在两个创新的高吞吐量筛选（HTS）范式中进行测定，以识别新颖抑制剂。一种方法将采用标准的长途道路来识别新型的小分子。我们将筛选 100,000种测定缓冲液中的化合物，其磷脂的调节是二聚化的。一个面板次要测定将用于表征已确认的HTS命中，以表征特异性，效力，作用方式和细胞活性。预期的目标1里程碑是识别适合提名的新铅作为一个临床前药物开发候选人。由于类似药物的小尺寸之间的不匹配分子与蛋白质蛋白相互作用（PPI）的额外表面积相比，HTS 蛋白质不满意的抑制剂受到挑战。为了加速将新疗法加速到诊所，我们提案一种更非常规的方法，我们将尝试确定两个FDA的组合通过变构协同作用的批准的药物，以防止自我关联和汇总作为。我们将使用开发的多重HTS方法来筛选FDA批准的药物的混合发现将单体稳定为结构的组合，从而防止自我关联。预期目标2 里程碑是确定两种先前批准的药物的组合概念验证临床测试比单个新化学实体更快地治疗突触核虫病。如果成功，我们开发的新方法是发现新颖的PPI调节器可能具有更广泛的实用性促进对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。