Discovery of novel USP8 inhibitors via Ubiquitin-Rho-110 fluorometric assay based high throughput screening

Jie Han; Yucheng Tian; Liang Yu; Qilin Zhang; Xi Xu; Yichao Zhang; Jubo Wang; Zengyi Ma; Jinlei Bian; Cheng Luo; Hualiang Jiang; Kaixian Chen; Yao Zhao; Zhiyu Li; Hong Ding

doi:10.1016/j.bioorg.2020.103962

Discovery of novel USP8 inhibitors via Ubiquitin-Rho-110 fluorometric assay based high throughput screening

Bioorg Chem. 2020 Aug:101:103962. doi: 10.1016/j.bioorg.2020.103962. Epub 2020 May 22.

Authors

Jie Han¹, Yucheng Tian², Liang Yu³, Qilin Zhang⁴, Xi Xu², Yichao Zhang⁴, Jubo Wang², Zengyi Ma⁴, Jinlei Bian², Cheng Luo³, Hualiang Jiang¹, Kaixian Chen³, Yao Zhao⁵, Zhiyu Li⁶, Hong Ding⁷

Affiliations

¹ 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.
² Jiangsu Key Laboratory of Drug Design and Optimization, Department of Medicinal Chemistry, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China.
³ State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, China.
⁴ Department of Neurosurgery, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
⁵ Department of Neurosurgery, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China. Electronic address: zhaoyaohs@vip.sina.com.
⁶ Jiangsu Key Laboratory of Drug Design and Optimization, Department of Medicinal Chemistry, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China. Electronic address: zhiyuli@cpu.edu.cn.
⁷ State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, China; Jiangsu Key Laboratory of Drug Design and Optimization, Department of Medicinal Chemistry, School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China. Electronic address: hding@simm.ac.cn.

PMID: 32480171
DOI: 10.1016/j.bioorg.2020.103962

Abstract

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 (IC₅₀) 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_D 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.

Keywords: Drug discovery; High throughput screening; USP8; Ubiquitination.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Endopeptidases
Endosomal Sorting Complexes Required for Transport / antagonists & inhibitors*
High-Throughput Screening Assays / methods*
Humans
Molecular Docking Simulation
Signal Transduction
Spectrometry, Fluorescence / methods
Surface Plasmon Resonance
Ubiquitin Thiolesterase / antagonists & inhibitors*
Ubiquitination

Substances

Endosomal Sorting Complexes Required for Transport
Endopeptidases
USP8 protein, human
Ubiquitin Thiolesterase