Development of selective substrate based inhibitors of ubiquitin isopeptidases

泛素异肽酶选择性底物抑制剂的开发

• 批准号: 8707677
• 负责人: Kumar Suresh
• 金额: $ 22.44万
• 依托单位: PROGENRA, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-15 至 2016-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8707677
• 关键词: Address; Biological Assay; Bortezomib; C-terminal; Cell Proliferation; Cells; Cleaved cell; Clinic; Clinical Trials; Development; Disease; Drug Industry; Enzymes; Failure; Family; Goals; Hematologic Neoplasms; Homeostasis; Human Genome; In Situ; In Vitro; Industry; Inflammation; Investigation; Label; Link; MDM2 gene; Malignant Neoplasms; Mediating; Multiple Myeloma; Pathway interactions; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Physiological; Physiological Processes; Play; Pre-Clinical Model; Proteasome Inhibitor; Proteins; Reagent; Reporter; Reporting; Role; Source; System; Testing; Therapeutic; Therapeutic Index; Toxic effect; Ubiquitin; Ubiquitin C; Ubiquitin-Activating Enzymes; Variant; Work; base; cancer cell; cancer therapy; drug discovery; enzyme pathway; enzyme therapy; fluorophore; high throughput screening; improved; in vivo; inhibitor/antagonist; innovation; isopeptidase; link protein; luciferin; meetings; multicatalytic endopeptidase complex; new therapeutic target; novel; pre-clinical; public health relevance; research clinical testing; scale up; screening; success; therapy development; ubiquitin C-terminal 7-amido-4-methylcoumarin; ubiquitin isopeptidase; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): The FDA approval of the proteasome inhibitor bortezomib/VELCADE(r) for the treatment of multiple myeloma and other hematological malignancies demonstrates that the ubiquitin pathway is a promising source of new drugs for the treatment of cancer and perhaps other diseases. Bortezomib, a proteasome inhibitor, exerts a global effect and its use is thus limited by a serious toxicity profile. Efforts are underway to ind ubiquitin pathway enzyme targets acting more selectively than the proteasome with the aim of developing drugs with improved toxicity profiles and therapeutic indices. Among these enzymes are the ubiquitin E1 activating enzyme, E2 conjugating enzyme, E3 ligase and Deubiquitylases (DUBs). DUBs act on limited numbers of target proteins and are linked biochemically and genetically to various diseases, including cancer. These enzymes are proteases and are considered druggable, and various pharmaceutical and biotech companies, including Progenra have identified novel, selective DUB inhibitors. Despite this progress, no DUB inhibitors have progressed to clinical evaluation. Part of the reason for this situation may be that improved screening assays are needed to identify hits for preclinical development that inhibit the DUB acting upon its in situ substrate. The project described in this application is the development of physiologically relevant substrate- based assay that introduces a significant improvement over all of the other assays currently in use to screen for DUB inhibitors. The assay is novel, homogeneous, HTS-compliant, and utilizes specific polyubiquitylated DUB substrates. The therapeutically relevant DUB/ubiquitylated substrate pairs USP7/HDM2/p53 (involved in cancer cell proliferation) will be prepared and purified, the assay configured, and a pilot screen for inhibitors will be conducted. This novel assay platform, when validated, will be expanded to include additional DUBs. The ultimate commercial goal of the proposed work is a robust, homogenous assay format that is suitable for HTS and customizable to various DUB/substrate pairs, allowing rapid progress in drug discovery for known and emerging DUB-mediated diseases.

描述（由申请人提供）：FDA批准蛋白酶体抑制剂硼替佐米/万珂（r）用于治疗多发性骨髓瘤和其他血液恶性肿瘤，表明泛素途径是治疗癌症和其他疾病新药的一个有前景的来源。硼替佐米是一种蛋白酶体抑制剂，具有全球效应，因此其使用受到严重毒性的限制。目前正在努力寻找比蛋白酶体更具选择性的泛素途径酶靶点，以开发具有改善的毒性特征和治疗指数的药物。这些酶包括泛素E1激活酶、E2结合酶、E3连接酶和去泛素化酶（DUBs）。DUBs作用于有限数量的靶蛋白，并在生物化学和遗传学上与包括癌症在内的各种疾病相关。这些酶是蛋白酶，被认为是可药用的，包括Progenra在内的各种制药和生物技术公司已经确定了新型的选择性DUB抑制剂。尽管取得了这一进展，但还没有DUB抑制剂进入临床评估阶段。这种情况的部分原因可能是需要改进的筛选试验来鉴定抑制DUB作用于其原位底物的临床前开发的命中物。本申请中描述的项目是开发基于生理学相关底物的测定法，其相对于目前用于筛选DUB抑制剂的所有其他测定法进行了显著改进。该测定是新颖的，均质的，符合HTS的，并利用特定的聚泛素化DUB底物。将制备和纯化治疗相关的DUB/泛素化底物对USP 7/HDM 2/p53（参与癌细胞增殖），配置测定，并进行抑制剂的中试筛选。这种新的检测平台在经过验证后，将扩展到包括其他DUB。拟议工作的最终商业目标是一种适用于HTS并可定制为各种DUB/底物对的稳健的均质测定形式，从而允许在已知和新兴DUB介导的疾病的药物发现方面取得快速进展。