Discussion

Authors: Afrah Sattikar, Keith Ansell, Lisa Hale, Peter Coombs, Tim Chapman, Claire Wallace, Kevin Gillen, Kristian Birchall, Craig Southern, Andy Merritt

Elevated levels of the cytokine Macrophage migration inhibitory factor (MIF) have been implicated in the pathogenesis of numerous autoimmune, metabolic and inflammatory diseases as well as cancer, making it an attractive drug target. Interestingly, MIF is a pleiotropic cytokine in that it also possesses enzymatic functions, namely tautomerase and oxidoreductase activities, although the physiologic function of these activities is unclear. Currently multiple small-molecule MIF inhibitors are in development, however the level of detailed published target validation using these inhibitors is limited. Therefore our goal has been to generate novel chemical starting points suitable for optimisation and characterisation in relevant cellular and in vivo disease models. Following an HTS of 80k compounds, a number of high priority hits were identified and assigned into structural clusters, which were subsequently were prioritised based on structure, biochemical activity and novelty. Here we describe the optimisation and application of the biochemical and biophysical assays that have been developed for hit validation and for support of the hit-to-lead chemistry program.