journal article·2026-07Open access

Discovery of a Potent Fluorescence Polarization Probe for Identifying USP1 Allosteric Inhibitors

Jiawei Cheng, Peipei Wang, Pengfei Wang, Jiamin Wang, Baiyang Li, Daqing Fang, Jian He, Xiaobei Hu, Weijuan Kan, Yubo Zhou, Chunpu Li, Jia Li, Hong Liu

Advanced Science· Volume 13· Issue 39

Abstract

ABSTRACT The deubiquitinating enzyme, ubiquitin‐specific protease 1 (USP1), is overexpressed in various tumor types, making it a promising target for cancer therapy. The development of USP1 allosteric inhibitors has progressed rapidly owing to their high selectivity and potency. However, the lack of appropriate chemical tools for developing a binding screen for this site has hampered the discovery of novel ligands. Herein, we developed the first allosteric fluoroprobe and fluorescence polarization (FP) assay for direct validation of USP1 allosteric inhibitors. The FP assay based on allosteric tracer 6‐2 enabled the differentiation of known allosteric and catalytic site inhibitors, providing a robust and scalable tool for high‐throughput screening. In addition, a novel class of potent tetrahydroisoquinoline USP1 inhibitors was identified, and the representative compound 14a possessed superior enzymatic and cellular activity compared with the clinical candidate KSQ‐4279, with potent in vivo anti‐DLBCL efficacy and good druggability. Collectively, this study provides a valuable fluoroprobe, FP assay platform, and lead compounds targeting USP1 for further structural optimization and antitumor mechanism studies.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: Ubiquitin and proteasome pathways · 14-3-3 protein interactions · Protein Degradation and Inhibitors

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Key Laboratory of Structure‐Based Drug Design and Discovery, Ministry of Education Shenyang Pharmaceutical University Shenyang China

State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai China

School of Chinese Materia Medica Nanjing University of Chinese Medicine Nanjing China

School of Pharmaceutical Science and Technology Hangzhou Institute for Advanced Study, UCAS Hangzhou China

University of Chinese Academy of Sciences Beijing China

Shandong Laboratory of Yantai Drug Discovery Bohai Rim Advanced Research Institute for Drug Discovery Yantai China

Air Force Hospital of Western Theater Command Chengdu China

Zhongshan Institute for Drug Discovery, Shanghai Institute of Materia Medica Chinese Academy of Sciences Zhongshan China

Funding

National Key Research and Development Program of China · National Natural Science Foundation of China · Shanghai Institute of Materia Medica, Chinese Academy of Sciences

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