Engineering 'Enzymelink' for screening lead compounds to inhibit mPGES-1 while maintaining prostacyclin synthase activity.

This study investigated our Enzymelinks, COX-2-10aa-mPGES-1 and COX-2-10aa-PGIS, as cellular cross-screening targets for quick identification of lead compounds to inhibit inflammatory PGE biosynthesis while maintaining prostacyclin synthesis. We integrated virtual and wet cross-screening using Enzymelinks to rapidly identify lead compounds from a large compound library. From 380,000 compounds virtually cross-screened with the Enzymelinks, 1576 compounds were identified and used for wet cross-screening using HEK293 cells that overexpressed individual Enzymelinks as targets. The top 15 lead compounds that inhibited mPGES-1 activity were identified. The top compound that specifically inhibited inflammatory PGE biosynthesis alone without affecting COX-2 coupled to PGI synthase (PGIS) for PGI biosynthesis was obtained. Enzymelink technology could advance cyclooxygenase pathway-targeted drug discovery to a significant degree.