Summary of in vitro studies.

Prednisolone (Pred) and Org 214007-0 (Org) were tested in different in vitro studies. A) GR binding: binding to recombinant human glucocorticoid receptor (GR) assessed by a fluorescence polarization competitor binding assay. Ki = inhibition constant or concentration of compound in the competitive binding assay which would occupy 50% of GR if no ligand was present. B) CHO – ind: Induction of gene expression measured in CHO cells stably co-transfected with human GR and a MMTV promoter – luciferase construct. C) HepG2 – ind: Induction of gene expression measured by microarray analysis of mRNA isolated from HepG2 cells incubated with either 1 μM prednisolone or 1 μM Org 214007-0. D) U2OS – rep: Repression of gene expression in U2OS cells overexpressing human GR. INFγ/TNFα – MCP-1 = IFNγ (100 ng/ml)/TNFα (50 ng/ml) induced MCP-1 release.IC50 or EC50 values represent the mean concentration of compound (±SD) required to resp. inhibit or effect the response to 50%. Maximal efficacy (Max. eff.) is expressed as the mean relative maximal effect (±SD) compared to the maximal effect by prednisolone (set at 100%). All assays (except for the micro array experiments) are performed at least two times.