Cancer drug discovery using disordered protein targets

使用无序蛋白质靶标发现癌症药物

• 批准号: 6690150
• 负责人: ALAN KEITH DUNKER
• 金额: $ 10万
• 依托单位: MOLECULAR KINETICS, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-05 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6690150
• 关键词: BCL2 gene /protein; SDS polyacrylamide gel electrophoresis; binding sites; drug discovery /isolation; elastases; intermolecular interaction; method development; molecular biology information system; neoplasm /cancer pharmacology; paclitaxel; polymerase chain reaction; protein binding; protein degradation; protein structure function; proteolysis; proteomics; time resolved data; trypsin

DESCRIPTION (provided by applicant): A novel technique for cancer drug discovery, "Disordered Protein Phage Display Assay", is proposed. This new technique will inject new avenues to cancer drug discovery methods since disordered proteins are only recently proven to have promising drug-targetable functions. The basic principle is based on the fact that disordered regions of proteins are more sensitive to protease digestion than ordered regions, whether naturally ordered or induced to be ordered. In Phase I, a model system using paclitaxel and bcl-2 will be used to demonstrate feasibility. This system is chosen because paclitaxel is a known anti-cancer drug and binds to the disordered loop of bcl-2. First, the hypothesis that small molecule (paclitaxel) binding to disordered target (bcl-2) inhibits its degradation by protease will be confirmed. Then the disordered region of bcl-2 will be cloned into a phage display vector and the hybrid will be tested by protease in the presence or absence of paclitaxel. Growth of the phage after protease and paclitaxel exposure indicates protection of the disordered region by paclitaxel. The ultimate final product will be a new methodology and expert analysis to accelerate drug lead identification via interaction with disordered regions of cancer-associated proteins.

描述（由申请人提供）：提出了一种用于癌症药物发现的新技术，“无序蛋白噬菌体展示测定”。这项新技术将为癌症药物发现方法注入新的途径，因为无序蛋白质最近才被证明具有有前途的药物靶向功能。其基本原理是基于这样的事实，即蛋白质的无序区域比有序区域对蛋白酶消化更敏感，无论是天然有序还是诱导有序。在第一阶段，使用紫杉醇和bcl-2的模型系统将用于证明可行性。之所以选择这个系统，是因为紫杉醇是一种已知的抗癌药物，并与bcl-2的无序环结合。首先，将证实小分子（紫杉醇）与无序靶（bcl-2）结合抑制其被蛋白酶降解的假设。然后将bcl-2的无序区克隆到噬菌体展示载体中，并在紫杉醇存在或不存在的情况下通过蛋白酶检测杂交体。蛋白酶和紫杉醇暴露后噬菌体的生长表明紫杉醇对无序区域的保护。最终的最终产品将是一种新的方法和专家分析，通过与癌症相关蛋白的无序区域相互作用来加速药物先导物的鉴定。