Developing Novel Arginine Biased Arrays for Epigenetic Research

Period of Performance: 08/01/2017 - 07/31/2018


Phase 2 SBIR

Recipient Firm

Epicypher, Inc.
Durham, NC 27713
Principal Investigator


PROJECT SUMMARY Epigenetic regulators represent a new frontier in drug discovery and are rapidly attracting pharmaceutical interests due to their remarkable ?druggability? and strong connection with human diseases (e.g., metabolic syndromes, neurodegenerative disorders, and cancers). To date, the epigenome consists of over 300 known proteins. However, very few epigenetic modifiers are being pursued in drug development programs due to our limited understanding of what proteins these enzymes regulate. This narrow saturation of the epigenetic ?drug arena? is fueled by a dearth of tools to systematically identify and characterize potential targets. To aid in the discovery of novel relationships between enzymes and the proteins that they target, we will commercialize high complexity oriented peptide array libraries, (OPALs). In Phase I studies, we synthesized OPAL libraries, comprising a core arginine (R- OPAL) or lysine (K- OPAL) residue with flanking degenerate sequences (+/- 3 or 4 amino acids in both N- and C- terminal directions; Figure 1) and demonstrated that OPALs are powerful tools to systematically characterize target motifs of writer enzymes. Our focus here is to develop OPALs to examine ?writer? enzymes that modify lysine or arginine residues. In Phase II, EpiCypher will scale up synthesis of the unmodified K- OPAL and R- OPAL peptide libraries and optimize a high- throughput 96- well assay format for validation using methyltransferase assays developed in the Phase I studies (Aim 1). We will also vastly expand the commercial utility of our discovery platform on two fronts: 1) developing a non- radioactive detection system and a user- friendly data analysis pipeline establishing (Aim 2)? and 2) expanding our K- OPAL platform to analyze lysine- acylation writer enzymes (Aim 3). Seven additional lysine- modified acyl group post- translational modifications (of which acetylation is a founding member) have been recently identified (e.g., crotonylation, butyrylation, or propionylation), which function together with lysine acetylation to regulate gene activation. Given the well- described role of histone acetylation in disease, these new acylation PTMs represent an exciting new field of epigenetic study that has great drug development potential. The innovative discovery platform described herein provides the first comprehensive screening tool to study epigenetic ?writer? enzymes. EpiCypher has assembled a strong scientific team consisting of pioneers in the field of epigenetic regulation (Drs. Mark Bedford and Scott Rothbart) and detection chemistry (Dr. Marcey Waters) to develop a robust, versatile, and user- friendly discovery platform that will have a powerful translational impact on epigenetic research and therapeutic development.