Inspiration from Fluorination: Chemical Epigenetics Approaches to Probe Molecular Recognition Events in Transcription
Protein-protein interaction inhibitor discovery has proven difficult due to the large surface area and dynamic interfaces of proteins. To facilitate the early lead discovery rate, I will first describe a rapid protein-based 19F NMR method for detecting protein-ligand interactions by screening low complexity molecules (fragments), drug-like molecules, and peptidomimetics. We have tested the sensitivity, accuracy, and speed of this method through screening libraries of small molecule fragments. The advantages of using 3D-fragments for discovery of more selective hits for bromodomain-containing proteins will be specifically highlighted. In the second part of the talk, I will describe improvements in our method for the field of epigenetics targeting bromodomain and extra-terminal (BET) family proteins. These studies have led to a selective inhibitor for the first bromodomain of BRD4. Structure-based design has identified several new design rules for maintaining selectivity and potency. Cellular efficacy in cancer and inflammatory model systems using this novel BRD4 inhibitor will be briefly described. Finally, development of a new heterocyclic scaffold for the second bromodomain of BRD4 will be highlighted. The speed, ease of interpretation, and low concentration of protein needed for binding experiments affords a new method to discover and characterize both native and new ligands for bromodomains and may find utility in the study of additional epigenetic “reader” domains.