State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, China; University of Chinese Academy of Sciences, 19 Yuquan Road, Beijing 100049, China.
Jiangsu Key Laboratory of Drug Design and Optimization, Department of Medicinal Chemistry, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China.
Bioorg Chem. 2020 Aug;101:103962. doi: 10.1016/j.bioorg.2020.103962. Epub 2020 May 22.
USP8, one member of deubiquitinating enzymes (DUBs) families, maintains the ubiquitination level of EGFR and regulates the downstream signaling pathways. The deregulation of USP8 has been implicated in many human diseases, especially in cancer. Therefore, USP8 has been identified as a promising target for drug design. Herein, via high throughput screening based on Ubiquitin-rhodamine-110 (Ubiquitin-Rho-110) fluorometric activity assay, we discovered a novel inhibitor DC-U43. By structure optimization, DC-U43-10 reached a half-maximal inhibitory concentration (IC) value of 2.6 ± 1.1 μM and exhibited 10-fold selectivity against USP7. The binding between DC-U43-10 and USP8 was validated by surface plasmon resonance (SPR) assay with a K value of 10.5 ± 3.7 μM. It also inhibited the colony formation of H1975 cells. Hence, DC-U43-10 represents a kind of USP8 inhibitors with novel scaffold and has broad prospects for being a probe for USP8-related academic and clinical research.
USP8 是去泛素化酶（DUBs）家族的一员，它维持 EGFR 的泛素化水平并调节下游信号通路。USP8 的失调与许多人类疾病有关，尤其是癌症。因此，USP8 已被确定为药物设计的一个有前途的靶点。在此，我们通过基于泛素-罗丹明-110（Ubiquitin-Rho-110）荧光活性测定的高通量筛选，发现了一种新型抑制剂 DC-U43。通过结构优化，DC-U43-10 的半最大抑制浓度（IC）值达到 2.6±1.1μM，并对 USP7 表现出 10 倍的选择性。表面等离子体共振（SPR）测定验证了 DC-U43-10 与 USP8 之间的结合，其 K 值为 10.5±3.7μM。它还抑制了 H1975 细胞的集落形成。因此，DC-U43-10 代表了一种具有新型支架的 USP8 抑制剂，具有广泛的用于 USP8 相关学术和临床研究的探针的前景。
