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Herein, we report the discovery of a series of new P300/CBP-associated factor (PCAF) bromodomain (BRD) inhibitors, which were obtained through a hit discovery process and subsequent structure-based optimization and structure-activity relationship analyses toward a retrieved hit compound (12). Among these inhibitors, ( R, R)-36n is the most potent one with an IC50 of 7 nM in homogeneous time-resolved fluorescence assay and a KD of 78 nM in isothermal titration calorimetry assay. This compound also exhibited activity against GCN5 and FALZ, but weak or no activity against other 29 BRD proteins and 422 kinases, indicating considerable selectivity. X-ray cocrystal structure analysis revealed the molecular interaction mode and the precise stereochemistry required for bioactivity. Cellular activity, preliminary RNA-seq analysis, and pharmacokinetic properties were also examined for this compound. Collectively, this study provides a versatile tool molecule to explore molecular mechanisms of PCAF BRD regulation and also offers a new lead compound for drug discovery targeting PCAF.

Original publication

DOI

10.1021/acs.jmedchem.9b00096

Type

Journal article

Journal

J Med Chem

Publication Date

09/05/2019

Volume

62

Pages

4526 - 4542

Keywords

Animals, Drug Discovery, Enzyme Inhibitors, Gene Expression, HEK293 Cells, Humans, Male, Mice, Microsomes, Liver, Molecular Structure, Protein Domains, Pyrimidinones, Pyrroles, Rats, Sprague-Dawley, Stereoisomerism, Structure-Activity Relationship, p300-CBP Transcription Factors