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: 7622287
• 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/7622287
• 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

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 attractivetherapeutics 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.

人们逐渐达成共识，认为非纤维状细胞内 Abeta 聚集，而不是不溶性原纤维， 是最有害的 Abeta 物种，可能在阿尔茨海默病 (AD) 中发挥重要作用 发病。因此，一种有吸引力的 AD 治疗方法是选择性降低 潜在的突触毒性 Abeta 通过稳定细胞内 Abeta 的单体形式或 使寡聚结构不稳定。低分子量药物代表最有吸引力的治疗方法 用于抑制 Abeta 聚集，因为许多小分子能够渗透血脑屏障 (BBB) 和穿过细胞膜。然而，从历史上看，蛋白质聚集一直是一种极其 合成类药物分子难以靶向，部分原因是其表面积通常较大 被两个相互作用的蛋白质覆盖，并且蛋白质之间有大而平坦的结合表面。其他 挑战在于，虽然新型有机化合物在测试中可能非常有效 实验室中的孤立目标，它们可能与所需细胞成分以外的细胞成分发生交叉反应 目标。自然界中发现的小分子，通常被称为“天然产物”，通常在 细胞在进化过程中并且不太可能以损害细胞的方式相互作用 膜或 DMA 等组件。此外，最近的研究表明，许多天然 产品在抑制多种蛋白质-蛋白质相互作用方面非常有效。因此，一个重要的 我们正在探索的问题是天然产物或类似天然产物的分子是否可以被分离出来 有效抑制 Abeta 聚集，同时被活细胞耐受。长期来看 本研究的目标是鉴定细胞内 Abeta 聚集的天然产物样抑制剂 作为治疗 AD 的治疗剂的潜力。为了实现这一目标，我们开发了一种基于细胞的检测方法 直接监测细胞内环境中的 Abeta 折叠。该特定应用旨在：(1) 配置我们新颖的基于细胞的折叠测定，用于组合小分子的高通量筛选 图书馆； (2) 从面向多样性的合成库中分离出类似天然产物的化合物，这些化合物是 能够拮抗Abeta聚集。此类化合物将作为 AD 治疗和 阐明 Abeta 折叠在介导神经毒性中的生理作用的生物学研究。