Preventing Tau uptake by novel inhibitors of tau binding to LRP1

通过与 LRP1 结合的新型 tau 抑制剂阻止 Tau 摄取

基本信息

批准号：
10343473
负责人：
Michael Jackson
金额：
$ 88.57万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA BARBARA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10343473
关键词：
Affect Affinity Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease pathology American Animal Model Antibodies Antisense Oligonucleotides Automation Binding Biochemical Biological Assay Biology Biophysics Brain Brain region Cells Cellular biology Chemicals Critical Pathways DNA Dementia Disease Disease Progression Dose Event Fluorescence Resonance Energy Transfer Future Goals Healthcare Systems Human Impaired cognition In Vitro Intervention LDL-Receptor Related Protein 1 Lead Learning Libraries Ligand Binding Domain Measures Mediating Mediator of activation protein Medical Morphologic artifacts Mus Neurodegenerative Disorders Neurofibrillary Tangles Neurons Pathogenesis Pathologic Pathology Pathway interactions Patients Pattern Penetration Performance Pharmaceutical Preparations Phenotype Powder dose form Probability Procedures Production Property Publishing Reagent Recombinant Proteins Series Site Societies Surface System Tauopathies Testing Validation Work analog base cheminformatics effective therapy high throughput screening in vivo Model in vivo evaluation induced pluripotent stem cell inhibitor insight knock-down member neurofibrillary tangle formation neurotoxicity new therapeutic target novel novel strategies novel therapeutic intervention novel therapeutics prevent protein aggregation prototype receptor response scaffold scale up screening small molecule small molecule libraries tau Proteins tau aggregation tau interaction tau mutation treatment strategy uptake

项目摘要

PROJECT SUMMARY Several neurodegenerative diseases, such as Alzheimer's disease (AD), are characterized by the spread and aggregation of the protein tau. Tau aggregates or neurofibrillary tangles (NFTs) accumulate throughout the brain of patients and lead to dementia. No effective treatments currently exist for tauopathies. Those approaches that have been considered, such as tau antibodies and antisense oligonucleotides, directly target tau. The complexity of tau cell biology, however, makes this approach challenging. A completely novel approach is to take advantage of the tau spreading pathway. The spread of NFTs correlate with disease progression and is a likely mediator for the observed neurotoxicity. Recently, we identified a cellular receptor, LRP1 (Low-density lipoprotein Receptor-related Protein 1), that regulates the tau spread pathway. Knockdown of LRP1 prevents tau spread in human iPS neurons and the mouse brain, suggesting that the tau-LRP1 interaction could be an important entry point for disease intervention. Therefore, the main objective of this project is to identify small-molecule chemical probes that prevent the binding of tau to LRP1, with the hypothesis that these molecules would serve as key starting points for novel therapeutics. In preliminary work, we have identified the primary interaction surface for tau on LRP1 and have developed a TR-FRET high-throughput screening (HTS) assay to identify compounds that can disrupt this interaction. We have optimized the assay to 1536-well format and conducted a pilot screen of 5,000 compounds with excellent performance, Z'~0.7 and a hit rate of ~0.4%. Analysis of the 20 hits from this screen in primary, artifact, and orthogonal assays identified several compounds with modest potency in dose response. To fully develop this work, we propose three aims. In Aim 1, we will use the TR-FRET assay to screen a 420,000 chemical library and, in parallel, conduct an affinity screen of a 4.4-billion- member DNA encoded library leveraging the DELopen platform (WuXi AppTec). In Aim 2, we will narrow our hit selection using orthogonal and novel biochemical profiling assays to determine mechanism of action. Finally, in Aim 3 we will validate hits with advanced biophysical and cell-based assays. We expect our multi-pronged approach will identify multiple chemical series with different mechanisms of action. Subsequent hit expansion efforts will produce chemical probes with properties suitable to test our hypothesis that small molecule LRP1-tau inhibitors can prevent tau uptake and spread. In future studies we intend to develop these probes into drugs that prevent tau spreading in tauopathies such as AD. As the critical path testing funnel is in place, we anticipate we can rapidly obtain and evaluate selective in vitro hits, explore their activity, and ultimately their suitability as starting points for hit-to-lead studies and for future in vivo evaluation in animal models and eventually patients.

项目概要一些神经退行性疾病，例如阿尔茨海默氏病 (AD)，其特点是传播和传播 tau 蛋白的聚集。 Tau 蛋白聚集体或神经原纤维缠结 (NFT) 在整个大脑中积聚并导致患者痴呆。目前尚无针对 tau 蛋白病的有效治疗方法。那些方法已经考虑过，例如 tau 抗体和反义寡核苷酸，直接靶向 tau。复杂性然而，tau 细胞生物学的复杂性使得这种方法具有挑战性。一种全新的方法是利用 tau 扩散途径。 NFT 的传播与疾病进展相关，并且可能是一个中介因素对于观察到的神经毒性。最近，我们鉴定了一种细胞受体，LRP1（低密度脂蛋白）受体相关蛋白 1)，调节 tau 扩散途径。敲低 LRP1 可防止 tau 扩散在人类 iPS 神经元和小鼠大脑中，表明 tau-LRP1 相互作用可能是一个重要的疾病干预的切入点。因此，该项目的主要目标是识别小分子阻止 tau 与 LRP1 结合的化学探针，假设这些分子可以发挥作用作为新疗法的关键起点。在前期工作中，我们已经确定了主要的相互作用 LRP1 上的 tau 表面，并开发了 TR-FRET 高通量筛选 (HTS) 测定法来识别可以破坏这种相互作用的化合物。我们已将测定优化为 1536 孔格式并进行了中试筛选了 5,000 种性能优异的化合物，Z'~0.7，命中率~0.4%。 20 分析在初级、伪影和正交测定中，从该屏幕中命中的结果鉴定出几种具有适度的化合物剂量反应的效力。为了充分开展这项工作，我们提出三个目标。在目标 1 中，我们将使用 TR-FRET 分析筛选 420,000 个化学库，同时对 44 亿个化学物质进行亲和力筛选利用DELopen平台（WuXi AppTec）的成员DNA编码库。在目标 2 中，我们将缩小目标范围使用正交和新颖的生化分析分析进行选择以确定作用机制。最后，在目标 3 我们将通过先进的生物物理和基于细胞的检测来验证命中。我们期待多管齐下方法将识别具有不同作用机制的多种化学系列。后续点击扩展我们的努力将产生具有适合检验我们假设的化学探针，即小分子 LRP1-tau 抑制剂可以阻止 tau 蛋白的摄取和扩散。在未来的研究中，我们打算将这些探针开发成药物防止 tau 蛋白在 AD 等 tau 病中扩散。随着关键路径测试漏斗的到位，我们预计我们可以快速获得和评估选择性体外命中，探索它们的活性，并最终确定它们的适用性命中先导研究以及未来动物模型和最终患者体内评估的起点。