Room-temperature fragment screening of soluble epoxide hydrolase by serial crystallography

Abstract

Room temperature serial crystallography offers advantages over conventional cryo-crystallography, such as simplified crystal handling and the possibility to avoid potential artefacts associated with cryo-trapping. However, to be considered as an alternative for drug discovery, where compound availability may be limited and speed of structure delivery is a key factor, it suffers from several limitations. To address these challenges, we have optimized a serial crystallography workflow for ligand soaking, data collection and data processing, significantly reducing time and reagent consumption to make it a viable option for drug discovery applications, herein exemplified by crystallographic fragment screening. Our approach incorporates the use of dried-in fragment cocktails on fixed target supports, compatible with 96-well plates for crystal soaking, and an efficient data processing pipeline tailored for serial crystallography. To validate our workflow, we conducted an in-crystal fragment screen at room temperature on the protein soluble epoxide hydrolase. The screen comprised 384 compounds and resulted in identification of 40 fragment binders corresponding to a hit rate of 10.4 %. The resulting room-temperature structures are of high quality and reveal opportunities for specific interaction within the highly hydrophobic active site of soluble epoxide hydrolase. Finally, we discuss potential avenues for further workflow optimization, highlighting the future potential of this approach for drug discovery. SynopsisWe have developed a workflow that allowed us to efficiently conduct a fragment screen at room temperature using serial crystallography, of interest for future drug discovery campaigns.