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：突触核蛋白病的新药物发现范例细胞蛋白质稳态功能障碍导致错误折叠蛋白质的异常积累，这些蛋白质与 α-突触核蛋白 (aS) 是多种神经退行性疾病的主要成分。称为路易体的细胞内包涵体——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) 的常见病理学可能与 aS 和 tau 病理学有关。由于这两种蛋白质可能发生核内和聚集，因此在 AD 中存在重叠。突触核蛋白病是弥合多种中枢神经系统蛋白质错误折叠疾病的高优先级目标。是一种 140 个氨基酸的细胞内蛋白质，本质上是无序单体，但可以采用多个 α-。与脂质囊泡结合时的螺旋构象是从无序到α-螺旋构象的转变。据认为会导致形成错误折叠的富含片层的结构，从而形成可溶性低聚物和我们的假设是膜磷脂诱导的 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，并且药物抑制；这一步将具有治疗价值。我们开发了一种离体分裂荧光素酶蛋白互补。我们的建议是使用此方法来检测 aS 寡聚和聚集时的初始二聚化。在两种创新的高通量筛选 (HTS) 范例中进行测定，以识别新型 aS 聚集一种方法将采取标准的漫长道路来识别新型小分子。测定缓冲液中的 100,000 种化合物经过磷脂调节，可增强 aS 二聚化。二次测定将用于表征已确认的 HTS 命中的特异性、效力、作用方式和预期的目标 1 里程碑是鉴定出适合提名为细胞活性的新先导化合物。临床前药物开发候选药物由于小尺寸之间的不匹配。分子与参与蛋白质-蛋白质相互作用 (PPI) 的广阔表面积相比，HTS 为了加速将新疗法推向临床，我们面临着挑战。提出一种更非常规的方法，我们将尝试确定两种 FDA 的组合批准通过变构协同作用发挥超添加剂作用，以防止自缔合和聚集我们将使用我们开发的多重 HTS 方法来筛选 FDA 批准的药物混合物发现稳定单体 aS 结构的组合，防止自缔合预期目标 2。里程碑是确定了两种先前批准的药物的组合，有可能推进到比单个新化学实体更快地治疗突触核蛋白病的概念验证临床测试。如果成功的话，我们开发的发现新型 PPI 调节剂的新方法可能具有更广泛的用途增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了解。