title: PAINS in the assay: chemical mechanisms of assay interference and promiscuous enzymatic inhibition observed during a sulfhydryl-scavenging HTS.
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.
authors: Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic College of Medicine , Rochester, Minnesota 55905, United States.
authors: Jayme L
authors: JL
authors: Dahlin
authors: J Willem M
authors: JW
authors: Nissink
authors: Jessica M
authors: JM
authors: Strasser
authors: Subhashree
authors: S
authors: Francis
authors: LeeAnn
authors: L
authors: Higgins
authors: Hui
authors: H
authors: Zhou
authors: Zhiguo
authors: Z
authors: Zhang
authors: Michael A
authors: MA
authors: Walters
mesh_terms: D001681:Biological Assay / Q000592:standards*; D015195:Drug Design; D055808:Drug Discovery; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D005189:False Positive Reactions; D057166:High-Throughput Screening Assays; D051548:Histone Acetyltransferases / Q000037:antagonists & inhibitors*; D006801:Humans; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular; D013438:Sulfhydryl Compounds / Q000737:chemistry*
chemical_list: D004791:Enzyme Inhibitors; D013438:Sulfhydryl Compounds; D051548:Histone Acetyltransferases
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