Summary of studies to define the therapeutic index (TI) of Org 214007-0.

A) THP-1: Microarray (m.a.) analysis of mRNA isolated from THP-1 cells incubated for 6 hours with either 1 μM prednisolone or 1 μM Org 214007-0 without (ind) or with IFNγ (220 ng/ml)/TNFα (374 ng/ml) ( = I/T), (rep) The mean percentage repression or induction of genes compared to that by prednisolone (set at 100%) is indicated. B) THP-1 – rep: Repression of gene expression in THP-1 cells. I/T – MCP-1, IL-6, IL-8 = TNFα (60 ng/ml)/IFNγ (40 ng/ml) induced MCP-1, IL-6 or IL-8 release. C) THP-1 – ind: Induction of FK506 binding protein 51 (FKBP51), glucocorticoid induced leucine zipper (GILZ) and dual specificity phosphatase 1 (DUSP1) in THP-1 cells. D) hWB – rep: Inhibition of LPS-induced (LPS) TNFα release or PMA/anti-CD28 (P/28) induced IL-5 release by primary human whole blood cells and hWB – ind: enhancement of PMA/anti-CD28/compound (P/28) induced G-CSF release by primary human whole blood cells. E) CASM3C – rep: Inhibition of MCP-1 release of coronary artery smooth muscle cells ( = CASM3C) stimulated with a cytokine mixture of IL-1β (1 ng/ml), IFNγ (100 ng/ml) and TFNα (5 ng/ml) ( = 1/I/T) by 1 μM prednisolone or 1 μM Org 214007-0. CASM3C – ind: Induction of serum amyloid A (SAA) of the cells mentioned above by 1 μM prednisolone or 1 M Org 214007-0. F) HDF3CGF – rep: inhibition of matrix metalloproteinase (MMP-1) release of human neonatal foreskin fibroblasts stimulated with the cytokine mixture mentioned above plus required growth factors ( = HDF3CGF). HDF3CGF – ind: Activation of plasminogen activator inhibitor-1 (PAI-1) by cells mentioned above by 1 μM prednisolone or 1 μM Org 214007-0. G) CIA mus.: Microarray (m.a.) analysis of mRNA isolated from muscle cells isolated at day 21, 2.5 hours after the final oral administration of either 1.5 mg/kg prednisolone or 0.3 mg/kg Org 214007-0 in the mouse CIA experiment. The mean percentage repression (rep) or induction (ind) of genes compared to that by prednisolone (set at 100%) is indicated.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%). A relative therapeutic index (TIrel) is calculated by the ratio of (the mean) % maximal efficacy in repression and (the mean) % maximal efficacy in induction of genes by Org 214007, where that of prednisolone is set at 1 (100%/100%). All assays (except for the microarray experiments) are performed at least two times.