title: Assessment of Bromodomain Target Engagement by a Series of BI2536 Analogues with Miniaturized BET-BRET.
abstract: Evaluating the engagement of a small molecule ligand with a protein target in cells provides useful information for chemical probe optimization and pharmaceutical development. While several techniques exist that can be performed in a low-throughput manner, systematic evaluation of large compound libraries remains a challenge. In-cell engagement measurements are especially useful when evaluating compound classes suspected to target multiple cellular factors. In this study we used a bioluminescent resonant energy transfer assay to assess bromodomain engagement by a compound series containing bromodomain- and kinase-biasing polypharmacophores based on the known dual BRD4 bromodomain/PLK1 kinase inhibitor BI2536. With this assay, we discovered several novel agents with bromodomain-selective specificity profiles and cellular activity. Thus, this platform aids in distinguishing molecules whose cellular activity is difficult to assess due to polypharmacologic effects.
authors.affiliation: Center for the Science of Therapeutics, Broad Institute, Cambridge, MA, 02142, USA.
authors.forename: Luke W
authors.initials: LW
authors.lastname: Koblan
authors.affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.|Department of Medicine, Harvard Medical School, Boston, MA, 02115, USA.
authors.forename: Dennis L
authors.initials: DL
authors.lastname: Buckley
authors.affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.|Department of Medicine, Harvard Medical School, Boston, MA, 02115, USA.|Center for the Science of Therapeutics, Broad Institute, Cambridge, MA, 02142, USA.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Ott
authors.affiliation: Center for the Science of Therapeutics, Broad Institute, Cambridge, MA, 02142, USA.|C4 Therapeutics, Cambridge, MA, 02142, USA.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Fitzgerald
authors.affiliation: Drug Discovery Department, Moffitt Cancer Center, Tampa, FL, 33612, USA.|Reaction Biology Corporation, Malvern, PA, 19355, USA.
authors.forename: Stuart W J
authors.initials: SW
authors.lastname: Ember
authors.affiliation: Drug Discovery Department, Moffitt Cancer Center, Tampa, FL, 33612, USA.
authors.forename: Jin-Yi
authors.initials: JY
authors.lastname: Zhu
authors.affiliation: Department of Chemistry, University of Massachusetts-Boston, Boston, MA, 02125, USA.
authors.forename: Shuai
authors.initials: S
authors.lastname: Liu
authors.affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.
authors.forename: Justin M
authors.initials: JM
authors.lastname: Roberts
authors.affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.
authors.forename: David
authors.initials: D
authors.lastname: Remillard
authors.affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.|Center for the Science of Therapeutics, Broad Institute, Cambridge, MA, 02142, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: Vittori
authors.affiliation: Department of Chemistry, University of Massachusetts-Boston, Boston, MA, 02125, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Drug Discovery Department, Moffitt Cancer Center, Tampa, FL, 33612, USA.
authors.forename: Ernst
authors.initials: E
authors.lastname: Schonbrunn
authors.affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.|Department of Medicine, Harvard Medical School, Boston, MA, 02115, USA.|Center for the Science of Therapeutics, Broad Institute, Cambridge, MA, 02142, USA.|Novartis Institutes for Biomedical Research, Cambridge, MA, 02139, USA.
authors.forename: James E
authors.identifier: http://orcid.org/0000-0002-2718-4415
authors.initials: JE
authors.lastname: Bradner
mesh_terms: D059447:Cell Cycle Checkpoints / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D015195:Drug Design; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D008163:Luminescent Measurements; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D011621:Pteridines / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000097869:Polo-Like Kinase 1; D000092468:Bromodomain Containing Proteins
chemical_list: D018797:Cell Cycle Proteins; D005456:Fluorescent Dyes; D009687:Nuclear Proteins; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D011621:Pteridines; D014157:Transcription Factors; D000097869:Polo-Like Kinase 1; C518477:BI 2536; C441289:BRD4 protein, human; D000092468:Bromodomain Containing Proteins
keywords: bromodomain inhibition; drug design; epigenetics; fluorescence; in-cell target engagement assays; luminescence