A cell-based screen for inhibitors of intracellular Abeta aggregation

基于细胞的细胞内 Abeta 聚集抑制剂筛选

• 批准号: 7680747
• 负责人: MATTHEW P DELISA
• 金额: $ 3.98万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7680747
• 关键词: Address; Alzheimer' s Disease; American; Amino Acid Sequence; Amyloid; Amyloid beta-Protein; Area; Arginine; Binding; Biological Assay; Biological Factors; Biological Response Modifier Therapy; Blood - brain barrier anatomy; Cell membrane; Cells; Cellular Assay; Consensus; DNA; Data; Development; Engineering; Environment; Equilibrium; Evolution; Genetic; Goals; In Vitro; Knowledge; Laboratories; Lead; Libraries; Life; Light; Mediating; Membrane; Methods; Molecular; Molecular Weight; Monitor; Nature; Nerve Degeneration; Neurodegenerative Disorders; Pathogenesis; Peptide Sequence Determination; Pharmaceutical Preparations; Physiological; Play; Proteins; Research; Role; Screening procedure; Solutions; Staging; Structure; Surface; Symptoms; System; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Thinking; Time; Twin Multiple Birth; Work; abeta accumulation; base; combinatorial; desire; genetic selection; high throughput screening; improved; in vivo; inhibitor/antagonist; innovation; neurotoxicity; novel; peptide P; protein aggregation; protein protein interaction; small molecule; small molecule libraries

DESCRIPTION (provided by applicant): There is an emerging consensus that non-fibrillar intracellular Abeta aggregates, rather than insoluble fibrils, are the most deleterious Abeta species and may play a central role in Alzheimer's disease (AD) pathogenesis. Thus, an attractive therapeutic approach to AD would be to seletively reduce the levels of potentially synaptotoxic Abeta aggregates by either stabilizing intracellular Abeta in its monomeric form or destabilizing the oligomeric structure. Low molecular weight drugs represent the most attractive therapeutics for inhibiting Abeta aggregation as many small molecules are capable of permeating the blood-brain barrier (BBB) and crossing cell membranes. Historically, however, protein aggregation has been an extremely difficult target to address with synthetic drug-like molecules, owing in part to the large surface area generally covered by two interacting proteins and to the large, flat binding surfaces between the proteins. Another challenge is that while new types of organic compounds may be extremely potent when tested against isolated targets in the laboratory, they may cross-react with cellular components other than the desired target. Small molecules found in nature, often called 'natural products', typically have spent time inside of a cell during the course of evolution and are less likely to interact in a manner that damages cellular components such as membranes or DMA. In addition, it has been shown recently that many natural products are quite effective at inhibiting a diverse array of protein-protein interactions. Thus, an important question that we are exploring is whether natural products or natural product-like molecules can be isolated that effectively inhibit Abeta aggregation and, at the same time, be tolerated by living cells. The long-term goal of this research is to identify natural product-like inhibitors of intracellular Abeta aggregation that have potential as therapeutic agents for treating AD. Towards this goal, we have generated a cell-based assay for directly monitoring Abeta folding in the intracellular environment. This particular application seeks to: (1) configure our novel cell-based folding assay for high-throughput screening of combinatorial small-molecule libraries; and (2) isolate natural product-like compounds from diversity-oriented synthesis libraries that are capable of antagonizing Abeta aggregation. Such compounds will serve as leads for AD therapy and for biological studies that illuminate the physiological role of Abeta folding in mediating neurotoxicity.

描述（由申请人提供）：有一个新兴的共识，即非细胞内Abeta骨料而不是不溶的原纤维是最有害的Abeta物种，并且可能在阿尔茨海默氏病（AD）发病机理中起核心作用。因此，一种有吸引力的AD治疗方法是通过稳定细胞内ABETA以其单体形式稳定或破坏寡聚结构的稳定性，将潜在的突触毒素ABETA聚集体的水平降低。低分子量药物是抑制ABETA聚集的最具吸引力的治疗药，因为许多小分子能够渗透到血脑屏障（BBB）和穿越细胞膜的情况下。然而，从历史上看，蛋白质聚集一直是与合成药物样分子解决的极其困难的靶标，部分原因是通常由两种相互作用的蛋白质覆盖的大表面积以及蛋白质之间的较大，平坦的结合表面。另一个挑战是，尽管针对实验室中的孤立靶标进行测试时，新型有机化合物可能会非常有效，但它们可能与所需靶标以外的其他细胞成分进行反应。在自然界中发现的小分子，通常称为“天然产物”，通常在进化过程中花费了时间在细胞内，并且以损害细胞成分（例如膜或DMA）的方式相互作用。此外，最近已经显示，许多天然产物在抑制各种蛋白质蛋白质相互作用方面非常有效。因此，我们正在探索的一个重要问题是，是否可以分离天然产物或天然产物样的分子，从而有效抑制Abeta聚集，同时可以通过活细胞耐受。这项研究的长期目标是鉴定具有细胞内Abeta聚集的天然产品样抑制剂，这些抑制剂具有治疗AD的治疗剂的潜力。为了实现这一目标，我们生成了一个基于细胞的测定法，用于直接监测细胞内环境中的Abeta折叠。该特定应用程序寻求：（1）配置我们的新型基于细胞的折叠测定法，以用于组合小分子库的高通量筛选； （2）能够拮抗Abeta聚集的多样性合成库中分离出天然产物样的化合物。这种化合物将用作AD疗法的铅和生物学研究，以阐明Abeta折叠在介导神经毒性中的生理作用。