Faculty, Staff and Student Publications

Publication Date

6-1-2025

Journal

SLAS Discovery

DOI

10.1016/j.slasd.2025.100233

PMID

40228580

PMCID

PMC12224672

PubMedCentral® Posted Date

7-3-2025

PubMedCentral® Full Text Version

Author MSS

Abstract

TRIP13, a promising target for cancer therapy, has been identified as a key regulator of the mitotic checkpoint. Overexpression of TRIP13 is associated with poor clinical outcomes in various cancers. Inhibition of TRIP13 has the potential to address therapeutic challenges in cancer, particularly in therapy-resistant and Rb-deficient cancers. Despite the potential therapeutic benefits of TRIP13 inhibition, the development of TRIP13 inhibitors has been hindered by the lack of a robust high-throughput screening (HTS) assay. We developed a luminescence-based biochemical assay for TRIP13 activity to address this challenge using the ADP-Glo detection system. This assay offers high sensitivity, low background signal, and ease of automation, making it ideal for HTS applications. A pilot screen of kinase-focused inhibitors library and a large-scale screen of 4000 additional compounds demonstrated the assay's robust performance with a z'-factor exceeding 0.85 and a signal-to-background (S/B) ratio near 6. From the 50 initial hits, rigorous validation identified anlotinib as the most potent TRIP13 inhibitor with an IC

Keywords

High-Throughput Screening Assays, Humans, Cell Cycle Proteins, ATPases Associated with Diverse Cellular Activities, Drug Discovery, Protein Kinase Inhibitors, Antineoplastic Agents, Neoplasms, Cell Line, Tumor, Drug Development, Quinolines

Published Open-Access

yes

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