Discovery of novel USP8 inhibitors via Ubiquitin-Rho-110 fluorometric assay based high throughput screening
Discovery of novel USP8 inhibitors via Ubiquitin-Rho-110 fluorometric assay based high throughput screening
复制标题
通过基于泛素-Rho-110 荧光测定的高通量筛选发现新型 USP8 抑制剂
DOI：
10.1016/j.bioorg.2020.103962
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发表时间：
2020
影响因子：
5.1
通讯作者：
Ding Hong
中科院分区：
文献类型：
--
作者：
Han Jie;Tian Yucheng;Yu Liang;Zhang Qilin;Xu Xi;Zhang Yichao;Wang Jubo;Ma Zengyi;Bian Jinlei;Luo Cheng;Jiang Hualiang;Chen Kaixian;Zhao Yao;Li Zhiyu;Ding Hong
USP 8是去泛素化酶（DUBs）家族的成员之一，维持EGFR的泛素化水平并调节下游信号通路。USP 8的失调与许多人类疾病，特别是癌症有关。因此，USP 8已被确定为药物设计的有希望的靶标。在此，通过基于Ubiquitin-Rho-110（Ubiquitin-Rho-110）荧光活性测定的高通量筛选，我们发现了一种新的受体DC-U43。通过结构优化，DC-U43- 10达到2.6 ± 1.1 μM的半数最大抑制浓度（IC 50）值，对USP 7表现出10倍的选择性。通过表面等离子体共振（SPR）分析验证了DC-U43 - 10与USP 8之间的结合，其Kd值为10.5 ± 3.7 μM。抑制H1975细胞的殖民地形成。因此，DC-U43- 10代表了一种具有新型支架结构的USP 8抑制剂，具有作为USP 8相关学术和临床研究探针的广阔前景。
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 inhibitorDC-U43. By structure optimization,DC-U43-10reached a half-maximal inhibitory concentration (IC50) value of 2.6 ± 1.1 μM and exhibited 10-fold selectivity against USP7. The binding betweenDC-U43-10and USP8 was validated by surface plasmon resonance (SPR) assay with aKDvalue of 10.5 ± 3.7 μM. It also inhibited the colony formation of H1975 cells. Hence,DC-U43-10represents a kind of USP8 inhibitors with novel scaffold and has broad prospects for being a probe for USP8-related academic and clinical research.