Identification of Small-Molecule Inhibitors of Protein-Protein Interactions

描述（由申请人提供）：最近鉴定蛋白质复合物和/或二元相互作用的高通量蛋白质组学研究不仅产生了丰富的信息，而且强调了蛋白质-蛋白质相互作用（PPIs）在细胞生物学中的普遍和基本作用。部分由于这些研究的结果，PPI作为具有巨大治疗潜力的有希望的药物靶点正在迅速获得认可，然而，对于PPI“可用药”的结构特征或构成PPI有效抑制剂的化学特性知之甚少。除了少数成功案例外，基于靶标的筛选策略在识别PPI抑制剂方面做得很差。本提案描述了一种开发PPIs小分子抑制剂的新方法。我们的方法将分子条形码技术与体内检测PPIs的两种既定方法相结合；酵母双杂交（Y2H）体系和蛋白质互补试验（PCA）。Y2H和PCA都是检测不同类型ppi的稳健方法，在这个意义上是互补的。它们在筛选和发现PPIs抑制剂方面的潜在效用已经被提出。通过利用分子条形码技术，斯坦福基因组技术中心（SGTC）在开发中发挥了先锋作用，以及SGTC目前广泛的化学筛选基础设施，我们建议筛选数百种与疾病相关的人类ppi，其中含有约20,000种不同的化合物。这些实验将产生一个庞大的数据集（代表数百万个单独的测试），这将是一个无与伦比的资源，用于解决关于PPI的可药物性的关键问题，以及定义PPI抑制剂的化学特征。通过这些筛选确定的特异性抑制剂也可能是精确解剖培养的人类细胞系中相关蛋白功能的有价值的生物学工具。此外，虽然它们不太可能具有治疗剂所需的药代动力学或药效学特性，但这些抑制剂可以作为新药的有希望的起点。

DESCRIPTION (provided by applicant): Recent high-throughput proteomic studies that identify protein complexes and/or binary interactions have not only produced a wealth of information, but have underscored the ubiquitous and fundamental role protein-protein interactions (PPIs) play in cell biology. In part as a consequence of these studies, PPIs are rapidly gaining acceptance as promising drug targets with tremendous therapeutic potential, however very little is known about the structural features that render a PPI "druggable", or the chemical characteristics that constitute an effective inhibitor of a PPI. With the exception of a small number of success stories, target-based screening strategies have done poorly at identifying PPI inhibitors. This proposal describes a novel approach to developing small-molecule inhibitors of PPIs. Our approach merges molecular bar-code technology with two established methodologies for detecting PPIs in vivo; the yeast two-hybrid (Y2H) system and the protein complementation assay (PCA). Both Y2H and PCA are robust methods that detect distinct types of PPIs, and in this sense are complementary. Their potential utility in screening for, and uncovering inhibitors of PPIs has been proposed previously. By leveraging the molecular bar-code technology, in which the Stanford Genome Technology Center (SGTC) has played a pioneering role in developing, as well as the extensive chemical screening infrastructure currently in place at the SGTC, we propose to screen hundreds of disease-associated human PPIs with ~20,000 diverse chemical compounds. These experiments will generate a massive dataset (representing millions of individual tests) that will be an unparalleled resource for addressing key questions regarding the druggability of PPIs, and the chemical features that define PPI inhibitors. Specific inhibitors identified by these screens could also be valuable biological tools for precisely dissecting the function of the associated proteins in cultured human cell lines. Furthermore, while they will not likely posses the pharmacokinetic or pharmacodynamic properties required of a therapeutic agent, these inhibitors could serve as promising starting points for new drugs. PUBLIC HEALTH RELEVANCE: Protein-protein interactions (PPIs) are a promising class of drug targets with wide-spread therapeutic potential. This proposal describes a unique merging of existing technologies with the goal of identifying novel chemical inhibitors of PPIs. These inhibitors could be valuable tools for studying biology and/or serve as starting points for developing new drugs.