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.

一种新的共识是，细胞内非纤维状的Abeta聚集，而不是不可溶的纤维， 是最有害的Abeta物种，可能在阿尔茨海默病(AD)中发挥核心作用 发病机制。因此，治疗阿尔茨海默病的一个有吸引力的方法是选择性地降低血浆中的 潜在的突触毒性Abeta聚集体通过稳定细胞内单体Abeta或 破坏低聚物结构的稳定性。低分子药物是最具吸引力的治疗药物 用于抑制Abeta聚集，因为许多小分子能够渗透血脑屏障 (BBB)和跨细胞膜。然而，从历史上看，蛋白质聚集是一种极端的 合成类药物分子很难达到目标，部分原因是通常情况下较大的表面积 被两个相互作用的蛋白质覆盖，并延伸到蛋白质之间的大而平的结合面。另一个 挑战在于，尽管新类型的有机化合物在测试时可能非常有效 在实验室中孤立的靶标，它们可能与所需的细胞成分以外的其他细胞成分发生交叉反应 目标。在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 组件，如膜或DMA。此外，最近的研究表明，许多天然的 产品在抑制多种蛋白质-蛋白质相互作用方面相当有效。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 这有效地抑制了Abeta的聚集，同时也被活细胞耐受。长期的 本研究的目标是确定具有天然产物样的细胞内Abeta聚集抑制物 作为治疗阿尔茨海默病的潜在药物。为了这个目标，我们已经产生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。这一特殊申请旨在：(1) 配置我们的新型基于细胞的折叠实验以高通量筛选组合小分子 文库；以及(2)从面向多样性的合成文库中分离天然产物类化合物 能够对抗Abeta聚集。这些化合物将作为AD治疗和治疗的先导 阐明Abeta折叠在介导神经毒性中的生理作用的生物学研究。