HTS of Microglia for Clearance of Amyloid-Beta in Alzheimer's Disease

描述（由申请人提供）：阿尔茨海默病（AD）的淀粉样蛋白假说提出，与疾病进展和认知下降相关的AD发病机制中的主要事件是淀粉样蛋白肽（A？）淀粉样前体蛋白的异常表达片段。因此，减缓AD进展的方法被设计为通过药理学抑制其合成或通过开发增加小胶质细胞对A？的清除的A？疫苗和抗体来防止A？在中枢神经系统（CNS）的积累。此外，疾病病理学中的继发性事件是由小胶质细胞响应于A？的激活而释放促炎细胞因子、趋化因子和神经毒性活性氧（ROS）和活性氮（RNS）引起的。虽然几种药物发现方法显示出抑制A？？由于药物累积，从这些方法中的许多方法衍生的候选药物具有仍然成问题的副作用。在这项SBIR研究中，我们将发现靶向未开发途径的药物，以促进AD脑中A？的减少并抑制相关的神经炎症。具体而言，我们将：1）开发使用人类小胶质细胞系的高通量测定以测量A？吞噬作用，并筛选已知药物、生物活性物质和药物样化合物的小分子文库以发现促进A？吞噬作用的化合物，2）开发测定以测量神经炎症介质的产生，并测试发现增强A？吞噬作用的化合物是否可以抑制A？的产生。在人小胶质细胞系和原代人小胶质细胞中诱导的神经炎症介质，和3）在细胞培养物中发现增强A β诱导的吞噬作用和抑制A β诱导的神经炎症介质的化合物，测试它们诱导原代人小胶质细胞清除A β的能力从AD患者获得的临床相关的死后脑组织。在该I期项目完成后，必要的AD测定、小分子干预的概念证明以及额外研究所需的专业知识将到位，以用于II期的先导候选药物的鉴定和优化。在美国，有450万人患有阿尔茨海默病（AD），每年的直接和间接护理费用估计为1000亿美元。这种疾病目前无法治愈，是住院的主要原因。家庭的情感代价是相当大的。我们的SBIR研究旨在探索新的途径，以达到有效的药物治疗AD。公共

DESCRIPTION (provided by applicant): The amyloid hypothesis of Alzheimer's disease (AD) proposes that the primary event in AD pathogenesis correlating with disease progression and cognitive decline is the accumulation of amyloid-¿ peptide (A¿?) an abnormally expressed fragment of the amyloid precursor protein. Accordingly, approaches to slow AD progression are designed to prevent central nervous system (CNS) accumulation of A¿, either by pharmacological inhibition of its synthesis or by development of A¿ vaccines and antibodies that augment clearance of A¿ by microglia. Moreover, a secondary event in disease pathology results from release of proinflammatory cytokines, chemokines, and neurotoxic reactive oxygen species (ROS) and reactive nitrogen species (RNS) by microglia in response to activation by A¿. Although several drug discovery approaches show promise in suppressing A¿ ??accumulation, drug candidates derived from many of these approaches have side effects that remain problematic. In this SBIR study, we will discover agents which target unexploited pathways to both promote the reduction of A¿ in AD brain and suppress the associated neuroinflammation. Specifically, we will: 1) Develop high throughput assays using human microglial cell lines to measure A¿ phagocytosis and screen small molecule libraries of known drugs, bioactives and drug-like compounds to discover compounds that promote A¿ phagocytosis, 2) Develop assays to measure production of mediators of neuroinflammation and test whether compounds found to augment A¿ phagocytosis can suppress production of A¿ induced mediators of neuroinflammation in human microglial cell lines and primary human microglia, and 3) Compounds found to augment A¿ induced phagocytosis and suppress A¿ induced mediators of neuroinflammation in cell culture are tested for their ablility to induce primary human microglia to clear A¿ from clinically relevant post-mortem brain tissues obtained from AD patients. After completion of this Phase I project, the necessary AD assays, proof of concept for small molecule intervention, and required expertise for additional studies will be in place for identification and optimization of lead drug candidates in Phase II. Project In the United States, 4.5 million people have Alzheimer's Disease (AD) where direct and indirect costs of care are estimated at $100 billion annually. The disease is currently not curable and is the leading cause of institutionalization. Emotional costs to families are considerable. Our SBIR studies are designed to probe new avenues to arrive at effective drug treatments for AD. Public