3 research outputs found
Structure-activity relationships for binding of 4-substituted triazole-phenols to macrophage migration inhibitory factor (MIF)
Macrophage migration inhibitory factor (MIF) is a versatile protein that plays a role in inflammation, autoimmune diseases and cancers. Development of novel inhibitors will enable further exploration of MIF as a drug target. In this study, we investigated structure-activity relationships of MIF inhibitors using a MIF tautomerase activity assay to measure binding. Importantly, we notified that transition metals such as copper (II) and zinc (II) interfere with the MIF tautomerase activity under the assay conditions applied. EDTA was added to the assay buffer to avoid interference of residual heavy metals with tautomerase activity measurements. Using these assay conditions the structure-activity relationships for MIF binding of a series of triazole-phenols was explored. The most potent inhibitors in this series provided activities in the low micromolar range. Enzyme kinetic analysis indicates competitive binding that proved reversible. Binding to the enzyme was confirmed using a microscale thermophoresis (MST) assay. Molecular modelling was used to rationalize the observed structure-activity relationships. The most potent inhibitor 2d inhibited proliferation of A549 cells in a clonogenic assay. In addition, 2d attenuated MIF induced ERK phosphorylation in A549 cells. Altogether, this study provides insights in the structure-activity relationships for MIF binding of triazole-phenols and further validates this class of compounds as MIF binding agents in cell-based studies
Structure-activity relationships for binding of 4-substituted
Macrophage migration inhibitory factor (MIF) is a versatile protein that plays a role in inflammation,
autoimmune diseases and cancers. Development of novel inhibitors will enable further exploration of
MIF as a drug target. In this study, we investigated structure-activity relationships of MIF inhibitors using
a MIF tautomerase activity assay to measure binding. Importantly, we notified that transition metals such
as copper (II) and zinc (II) interfere with the MIF tautomerase activity under the assay conditions applied.
EDTA was added to the assay buffer to avoid interference of residual heavy metals with tautomerase
activity measurements. Using these assay conditions the structure-activity relationships for MIF binding
of a series of triazole-phenols was explored. The most potent inhibitors in this series provided activities
in the low micromolar range. Enzyme kinetic analysis indicates competitive binding that proved
reversible. Binding to the enzyme was confirmed using a microscale thermophoresis (MST) assay. Mo�lecular modelling was used to rationalize the observed structure-activity relationships. The most potent
inhibitor 2d inhibited proliferation of A549 cells in a clonogenic assay. In addition, 2d attenuated MIF
induced ERK phosphorylation in A549 cells. Altogether, this study provides insights in the structure�activity relationships for MIF binding of triazole-phenols and further validates this class of compounds
as MIF binding agents in cell-based studies
Structure–Activity Relationships and Anti-inflammatory Activities of <i>N</i>‑Carbamothioylformamide Analogues as MIF Tautomerase Inhibitors
Macrophage migration inhibitory factor
(MIF), a proinflammatory
cytokine, is an attractive therapeutic target for the treatment of
inflammatory diseases. In our previous study, 3-[(biphenyl-4-ylcarbonyl)Âcarbamothioyl]Âamino
benzoic acid (compound <b>1</b>) was discovered as a potent
inhibitor of MIF by docking-based virtual screening and bioassays.
Here, a series of analogues of compound <b>1</b> derived from
similarity search and chemical synthesis were evaluated for their
MIF tautomerase activities, and their structure–activity relationships
were then analyzed. The most potent inhibitor (compound <b>5</b>) with an IC<sub>50</sub> of 370 nM strongly suppressed lipopolysaccharide
(LPS)-induced production of TNF-α and IL-6 in a dose-dependent
manner and significantly enhanced the survival rate of mice with LPS-induced
endotoxic shock from 0 to 35% at 0.5 mg/kg and to 45% at 1 mg/kg,
highlighting the therapeutic potential of the MIF tautomerase inhibition
in inflammatory diseases