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

基本信息

批准号：
8018614
负责人：
Geoffrey Myles Wahl
金额：
$ 4.71万
依托单位：
SALK INSTITUTE FOR BIOLOGICAL STUDIES
依托单位国家：
美国
项目类别：
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/8018614
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 Future 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 follow-up genetic profiling 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

项目摘要

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 例归因于该疾病。肿瘤样本的遗传和分子分析已鉴定出可被小分子靶向以获得治疗益处的蛋白质。 p53 肿瘤抑制途径就是这样的一个例子。 p53 基因仍然是野生型，但该蛋白质在大约 50% 的人类癌症中功能失活。在这些情况下，经常观察到 p53 的主要负调节因子 mdm2 和 mdmx 的过度表达。 Mdm2 是一种 E3 泛素连接酶，催化泛素从 E2 泛素结合酶转移至 p53。这反过来又导致 p53 的蛋白酶体依赖性降解。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 mdm2 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 通过所谓的 RING 结构域相互作用，产生最有效的 E3 连接酶活性。因此，鉴定能够抑制 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方法。该提案的具体目标描述了基于细胞的 MLCPN 小分子库筛选，以发现 mdm2/mdmx RING-RING 相互作用的小分子抑制剂。其他后续研究将验证“命中”的特异性及其在表达野生型 p53 的肿瘤细胞中的功效。这些研究的成功完成将证明小分子可以抑制 mdm2/mdmx 相互作用，从而稳定和激活 p53。该筛选中主要命中的化学修饰将为基础科学研究产生有价值的化学探针，以及可用于药物开发的潜在先导化合物。公共卫生相关性：在美国，癌症在死亡原因中仅次于心血管疾病，位居第二，并且是全世界的头号杀手。 p53 肿瘤抑制因子是野生型，但在约 50% 的所有癌症中功能失活，并且是小分子抗癌药物重新激活的有吸引力的靶标。该提案的目的描述了一种鉴定一类新型 p53 激活化合物的方法，其成功完成将对未来携带野生型 p53 的癌症的治疗产生影响。