Virtual Screening of Some Active Human Macrophage Migration Inhibitory Factor Antagonists

Abstract

Macrophage migration inhibitory factor (MIF) is an autocrine- and paracrine-acting cytokine that is involved in several inflammatory, autoimmune, infectious, and oncogenic diseases. Clinical data have shown that inhibition of MIF, especially its tautomerase activity, with small compounds has been beneficial in some disease models. A virtual screening (VS) experiment is conducted for searching some active compounds to inhibit the tautomerase activity of MIF from the ZINC database. By using an x-ray-determined structure OXIM-11 as the query and an in-house developed two-dimensional scaffold comparing method designated as Sfilter, we have screened out some 1500 compounds for ranking by our previously published docking method ADDock. After further ranking by ADDock on 119 compounds screened, we have decided to choose 17 of them for measuring their inhibitory activity IC50 against the MIF tautomerase experimentally. The IC50's are measured using both human monocytic THP-1 cell lysate and purified recombinant human MIF protein. We have found that the IC50's measured for three searched compounds (namely, ZINC02693801, ZINC00141102, and ZINC12368346) are better than that determined for ISO-1, a known MIF tautomerase inhibitor and standard used throughout our VS experiment. Moreover, the scaffolds of most of our active compounds searched are also quite different from those searched and published by others previously.

Novel half‐life extended anti‐MIF nanobodies protect against endotoxic shock

“…Whereas several small‐molecule inhibitors of the biologic activity of MIF have been documented (65, 69, 70), to our knowledge, this is the first report to establish that an MIF antagonist, when appropriately engineered, can prevent lethal endotoxemia when administered as a single injection. Although monoclonal/polyclonal anti‐MIF Abs were also found to exert protective effects (34, 35, 71), Nbs have an advantage in that they can be tailored into multifunctional formats as demonstrated herein.…”

A Fluorescence Polarization Assay for Binding to Macrophage Migration Inhibitory Factor and Crystal Structures for Complexes of Two Potent Inhibitors

“…Most studies to identify MIF inhibitors have screened compound libraries using the DOPA or HPP tautomerase assays. 4 , 9 , 11 − 14 IC 50 or K i values are reported for inhibition of the tautomerization of these substrates. As discussed previously, 15 execution of these assays is complicated by multiple factors including the light sensitivity of DOPA, the slow rate of tautomerization of HPP, spectral interference of inhibitors and products, choice of protein concentration, and short times for the linear range of product formation in both cases.…”

Design, Synthesis, and Protein Crystallography of Biaryltriazoles as Potent Tautomerase Inhibitors of Macrophage Migration Inhibitory Factor

“…7 For example, this has been demonstrated through assay results for tautomerase activity, MIF/CD74 binding, and MIF-induced phosphorylation of ERK1/2 in inflamed cells, production of interleukins, glucocorticoid overriding ability, and macrophage chemotactic migration. 7–12 Nevertheless, the discovery of potent MIF tautomerase inhibitors is not well advanced as most inhibitors have arisen from screening exercises with no lead optimization. 8,11,12 In our efforts, 8–10 lead optimization has been limited by the lack of crystal structures for our tautomerase inhibitors bound to MIF, associated inconsistency between modeling results and activity data, and sensitivity of assay results to the substrate and protein source.…”

“…7–12 Nevertheless, the discovery of potent MIF tautomerase inhibitors is not well advanced as most inhibitors have arisen from screening exercises with no lead optimization. 8,11,12 In our efforts, 8–10 lead optimization has been limited by the lack of crystal structures for our tautomerase inhibitors bound to MIF, associated inconsistency between modeling results and activity data, and sensitivity of assay results to the substrate and protein source. As described here, these issues have been overcome for the series of biaryltriazoles, 1–3 .…”