PAINS in the assay: Chemical mechanisms of assay interference and promiscuous enzymatic inhibition observed during a sulfhydryl-scavenging HTS

Jayme L. Dahlin, J. Willem M. Nissink, Jessica M. Strasser, Subhashree Francis, Leeann Higgins, Hui Zhou, Zhiguo Zhang, Michael A. Walters

Research output: Contribution to journalArticle

140 Scopus citations

Abstract

Significant resources in early drug discovery are spent unknowingly pursuing artifacts and promiscuous bioactive compounds, while understanding the chemical basis for these adverse behaviors often goes unexplored in pursuit of lead compounds. Nearly all the hits from our recent sulfhydryl-scavenging high-throughput screen (HTS) targeting the histone acetyltransferase Rtt109 were such compounds. Herein, we characterize the chemical basis for assay interference and promiscuous enzymatic inhibition for several prominent chemotypes identified by this HTS, including some pan-assay interference compounds (PAINS). Protein mass spectrometry and ALARM NMR confirmed these compounds react covalently with cysteines on multiple proteins. Unfortunately, compounds containing these chemotypes have been published as screening actives in reputable journals and even touted as chemical probes or preclinical candidates. Our detailed characterization and identification of such thiol-reactive chemotypes should accelerate triage of nuisance compounds, guide screening library design, and prevent follow-up on undesirable chemical matter.

Original languageEnglish (US)
Pages (from-to)2091-2113
Number of pages23
JournalJournal of Medicinal Chemistry
Volume58
Issue number5
DOIs
StatePublished - Mar 12 2015

ASJC Scopus subject areas

  • Molecular Medicine
  • Drug Discovery

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    Dahlin, J. L., Nissink, J. W. M., Strasser, J. M., Francis, S., Higgins, L., Zhou, H., Zhang, Z., & Walters, M. A. (2015). PAINS in the assay: Chemical mechanisms of assay interference and promiscuous enzymatic inhibition observed during a sulfhydryl-scavenging HTS. Journal of Medicinal Chemistry, 58(5), 2091-2113. https://doi.org/10.1021/jm5019093