High throughput screen for inhibitors of the mdm2/mdmx interaction

mdm2/mdmx 相互作用抑制剂的高通量筛选

• 批准号: 7844780
• 负责人: Geoffrey Myles Wahl
• 金额: $ 4.76万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7844780
• 关键词: Amino Acid Sequence; Antineoplastic Agents; Behavior; Binding; C-terminal; Cardiovascular Diseases; Cause of Death; Cells; Cessation of life; Chemicals; Complex; Data; Development; Disease; Drug Delivery Systems; Exhibits; Follow-Up Studies; Future; Genetic; Growth; Homologous Protein; Human; In Vitro; Kinetics; Lead; Malignant Neoplasms; Mediating; Modification; Molecular Bank; Molecular Profiling; N-terminal; Pathway interactions; Pharmaceutical Preparations; Protein p53; Proteins; Research; Sampling; Screening procedure; Specificity; TP53 gene; Therapeutic; Ubiquitin; Ubiquitin-Conjugating Enzymes; United States; United States National Institutes of Health; base; cancer therapy; chemotherapy; drug development; drug discovery; high throughput screening; inhibitor/antagonist; multicatalytic endopeptidase complex; neoplastic cell; novel; novel strategies; overexpression; public health relevance; repository; small molecule; small molecule libraries; tumor; ubiquitin ligase; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): Cancer is the second leading cause of death in the United States: 1 in every 4 deaths is attributable to the disease. Genetic and molecular profiling of tumor samples has lead to the identification of proteins that may be targeted by small molecules for therapeutic benefit. One such example is the p53 tumor suppressor pathway. The p53 gene remains wild type, yet the protein is functionally inactivated in approximately 50% of all human cancers. In these cases, overexpression of mdm2 and mdmx, the major negative regulators of p53, is frequently observed. Mdm2 is an E3 ubiquitin ligase, and catalyzes the transfer of ubiquitin from E2 ubiquitin conjugating enzyme(s) to p53. This in turn leads to proteasome-dependent degradation of p53. Many drug discovery efforts have focused on disruption of the mdm2/p53 interaction in order to increase p53 activity in tumors. However, current mdm2-specific antagonists cannot disrupt the interaction of mdmx with p53. Furthermore, recent data indicate that the interaction of mdm2 with mdmx, via so-called RING domains, engenders the most effective E3 ligase activity. Therefore, the identification of small molecules that can inhibit the mdm2/mdmx interaction may provide a new approach to p53 stabilization and activation in cancer. The Specific Aims of this proposal describe a cell-based screen of the MLCPN small molecule library to discover small molecule inhibitors of the mdm2/mdmx RING-RING interaction. Additional follow-up studies are presented that will validate the specificity of 'hits', and their efficacy in tumor cells that express wild type p53. The successful completion of these studies would provide proof-of-principle that small molecules can inhibit the mdm2/mdmx interaction, leading to stabilization and activation of p53. The chemical modification of primary hits in this screen will generate valuable chemical probes for basic scientific research, as well as potential lead compounds that may be taken forward for drug development. PUBLIC HEALTH RELEVANCE: Cancer ranks second to cardiovascular diseases in causes of death in the United States, and is a leading killer worldwide. The p53 tumor suppressor is wild type but functionally inactivated in ~50% of all cancers, and is an attractive target for reactivation by small molecule anticancer drugs. The Aims of this proposal describe an approach to identify a novel class of p53 activating compounds, and their successful completion will have implications for future treatment of cancers harboring wild type p53.

描述（由申请人提供）：癌症是美国第二大死亡原因：每4例死亡中就有1例可归因于该疾病。肿瘤样本的遗传和分子分析已经导致鉴定出可能被小分子靶向治疗的蛋白质。其中一个例子就是p53肿瘤抑制通路。p53基因仍然是野生型，但在大约50%的人类癌症中，这种蛋白质在功能上失活。在这些情况下，经常观察到p53的主要负调节因子mdm2和mdmx的过表达。Mdm2是E3泛素连接酶，并催化泛素从E2泛素偶联酶转移到p53。这反过来导致蛋白酶体依赖性的p53降解。许多药物发现工作都集中在破坏mdm2/p53相互作用，以增加肿瘤中p53的活性。然而，目前的mdmx特异性拮抗剂不能破坏mdmx与p53的相互作用。此外，最近的数据表明，mdm2与mdmx的相互作用，通过所谓的环结构域，产生最有效的E3连接酶活性。因此，发现能够抑制mdm2/mdmx相互作用的小分子可能为癌症中p53的稳定和激活提供新的途径。本提案的具体目的描述了基于细胞的MLCPN小分子文库筛选，以发现mdm2/mdmx RING-RING相互作用的小分子抑制剂。进一步的后续研究将验证“命中”的特异性，以及它们对表达野生型p53的肿瘤细胞的有效性。这些研究的成功完成将提供小分子可以抑制mdm2/mdmx相互作用，从而稳定和激活p53的原理证明。对该筛选中的初级靶点进行化学修饰，将为基础科学研究产生有价值的化学探针，以及可能用于药物开发的潜在先导化合物。