Blacklow, Stephen C.Bradner, JamesShaw, Katharin2019-08-092017-052017-05-162017Shaw, Katharin. 2017. A Chemical Biology Platform to Develop Small Molecule Probes for Bromodomains. Doctoral dissertation, Harvard University, Graduate School of Arts & Sciences.http://nrs.harvard.edu/urn-3:HUL.InstRepos:41142078Bromodomain epigenetic reader proteins play crucial roles in the pathogenesis of diseases ranging from inflammation to cancer. Small molecule inihibitors have engendered a nuanced understanding of the role of BET subfamily bromodomains in multiple diseases. Here, platforms were established to facilitate chemical probe development for the remaining ~50 bromodomains with a concentrated focus on establishing a pipeline for the discovery of CBP bromodomain inhibitors. CBP is a large bromodomain-containing transcriptional co-activator involved in cell proliferation and growth, cellular differentiation and development, cognitive function, and cell adhesion. It is involved in the pathogenesis of cancer, in particular hematologic diseases. A high-throughput biochemical assay, cellular viability assay, and cellular target engagement assay were established in order to screen and validate small molecule inhibitors of the CBP bromodomain. A family-wide bromodomain profiling platform (BROMOscan) was established and fully validated to provide broad quantitative binding affinity and selectivity information. Using this platform, novel small molecule inhibitors for non-BET domains were discovered. Moreover, a LRRK2 kinase inhibitor with potent bromodomain cross-reactivity was identified and optimized. Using BROMOscan and the CBP bromodomain assays deeloped here, a dually functioning kinase-bromodomain inhibitor was discovered that was potent and selective for CBP.application/pdfenchemical biologyassay developmentbromodomainsepigeneticsinhibitionsmall moleculesThesis or Dissertation2019-08-09