Development of ubiquitin-specific protease 8 (USP8) inhibitors

泛素特异性蛋白酶 8 (USP8) 抑制剂的开发

• 批准号: 10460124
• 负责人: Anthony Xavier Ayala
• 金额: $ 4.19万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10460124
• 关键词: Attention; Autophagocytosis; Binding; Biological Assay; Biological Models; Biology; Biomedical Research; Cancer Biology; Cancer cell line; Caspase; Cell Line; Cells; Cellular Assay; Crystallography; DNA Sequence Alteration; Dana-Farber Cancer Institute; Development; Down-Regulation; Drug Targeting; Drug resistance; ERBB2 gene; ERBB3 gene; Endocytosis; Enzymes; Epidermal Growth Factor Receptor; Excision; Future; Gefitinib; Generations; Genetic; Goals; Growth Factor Receptors; Human Pathology; Institutes; Lead; Libraries; Lysosomes; MAP Kinase Gene; Malignant neoplasm of lung; Mass Spectrum Analysis; Measures; Mediating; Mutation; Non-Small-Cell Lung Carcinoma; Oncogenic; Oncology; Patients; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Phenotype; Play; Positioning Attribute; Property; Proteins; Proteome; Proteomics; Proto-Oncogene Proteins c-akt; Public Health Applications Research; Receptor Protein-Tyrosine Kinases; Reporting; Research; Resistance; Resistance development; Role; Sampling; Signal Pathway; Signaling Molecule; Small Interfering RNA; Solubility; Specificity; Surveys; System; Therapeutic; Time; USP6 gene; Ubiquitin; Ubiquitin Specific Protease 8; Ubiquitination; Universities; Validation; analog; base; cancer therapy; cellular targeting; design; drug discovery; experimental study; high throughput screening; improved; in vivo Model; inhibitor; inhibitor therapy; knock-down; lead optimization; lung cancer cell; multidisciplinary; mutant; novel; novel strategies; programs; protein profiling; scaffold; structural biology; targeted treatment; therapeutic target; tool; transcriptomics; ubiquitin-specific protease

Project Summary Ubiquitin specific protease 8 (USP8) is a cysteine protease that can deubiquinate receptor tyrosine kinases. USP8 regulates endocytosis and through this activity modulates the degradation of diverse substrates by the autophagy- lysosome system. In oncology, USP8 has been shown to deubiquinate and stabilize mutant epidermal growth factor receptor (EGFR) in non-small cell lung cancer (NSCLC). Preliminary studies show that USP8 inhibition promotes ubiquitin-mediated degradation of mutant EGFR in NSCLC cell lines and primary patient samples both sensitive and resistant to EGFR kinase inhibition. These results suggest that UPS8 inhibition could provide a therapeutic strategy for overcoming gefitinib-resistance in NSCLC. However, the USP8 inhibitor used in these preliminary studies is an early generation inhibitor that lacks the requisite potency and selectivity of a high-quality probe compound. The goal of this research program is to develop the first reversible USP8 inhibitors with the requisite potency, selectivity, and cell permeability to interrogate the potential of USP8 as a therapeutic target in NSCLC. The first aim is to perform medicinal chemistry optimization of lead scaffolds identified from a high throughput screen. I will design analogs of each compound to broadly survey regions of the molecule that are required for activity and positions that can be modified to modulate properties such as solubility and stability. For these studies, one position in the molecule will be modified at a time then productive changes will be combined. In the second aim, these inhibitors will be characterized with respect to their cellular target engagement, anti-proliferative effects, and effects on the stability of EGFR in mutant and wild type NSCLC cell lines. In the third aim, I will use the newly developed USP8 inhibitors as probes in phenotypic, proteomic, and transcriptomic assays across a broad set of cancer cell lines to uncover new USP8 biology in an unbiased manner. This proposal therefore outlines the development of the first USP8 inhibitors with the required potency and selectivity to be used as cellular probes to study USP8 biology, and contributes to evidence for USP8 inhibition as a therapeutic strategy for NSCLC.

项目摘要 泛素特异性蛋白酶8（USP 8）是一种半胱氨酸蛋白酶，可以使受体酪氨酸激酶去泛素化。USP8 调节内吞作用，并通过这种活性调节自噬对不同底物的降解， 溶酶体系统在肿瘤学中，USP8已被证明可以去泛素化并稳定突变的表皮生长 因子受体（EGFR）在非小细胞肺癌（NSCLC）。初步研究表明，USP8抑制 促进NSCLC细胞系和原发性患者样本中泛素介导的突变型EGFR降解， 对EGFR激酶抑制敏感且耐药。这些结果表明，UPS8抑制可以提供一种新的治疗方法。 克服NSCLC吉非替尼耐药性的治疗策略。然而，这些中使用的USP8抑制剂 初步研究是一个早期的一代抑制剂，缺乏必要的效力和选择性的高质量的 探针化合物。 这项研究计划的目标是开发第一个具有必要效力的可逆USP8抑制剂， 选择性和细胞渗透性，以探讨USP 8作为NSCLC治疗靶点的潜力。第一 目的是对从高通量筛选中鉴定的先导支架进行药物化学优化。我 将设计每种化合物的类似物，以广泛调查活性所需的分子区域， 可以被修饰以调节诸如溶解度和稳定性的性质的位置。在这些研究中，1 分子中的位置将被修改，然后将组合产生的变化。第二个目标， 这些抑制剂的特征在于它们的细胞靶结合，抗增殖作用， 以及对突变型和野生型NSCLC细胞系中EGFR稳定性的影响。在第三个目标中，我将使用 新开发的USP8抑制剂在表型、蛋白质组学和转录组学检测中作为探针， 一组癌细胞系，以公正的方式揭示新的USP8生物学。因此，本提案概述了 开发第一个具有所需效力和选择性的USP8抑制剂，用作细胞探针 研究USP 8生物学，并为USP 8抑制作为NSCLC治疗策略提供证据。