Fig. 2: CT-179 specificity analysis and pharmacokinetics analysis.
From: Suppressing recurrence in Sonic Hedgehog subgroup medulloblastoma using the OLIG2 inhibitor CT-179
A RCA in replicate HEK-293 cells studied by FCCS. Negative control (NC) cells were co-transfected with eGFP and Tomato (n = 15). Cells transfected with OLIG2-eGFP and OLIG2-Tomato showed increased RCA. Treatment with 1 µM CT-179 for one hour significantly decreased RCA in these cells. (n = 22 untreated, 19 CT-179-treated). B Dose-response curve showing RCA measured as in A, plotted against CT-179 concentration. Graph shows means ± SD for replicate cells (biological replicates, n = 15–22 as shown). Best fit of dose-response curve (red solid line) determined the half maximal inhibitory concentration of CT-179, IC50 = 1250 nM, and the allosteric factor, nallo = −0.66. C OLIG2-eGFP brightness, in counts per second per molecule (CPM) in untreated cells (n = 17) and in cells treated with 1 μM (n = 20) or 10 μM CT-179 (n = 17). D Diffusion coefficient (DC) of DNA-binding component in indicated conditions n = 16 untreated cells, 20 cells with 1 μM CT-179, 16 cells with 10 μM CT-179. CT-179 increased diffusion coefficients, reflecting increased OLIG2 mobility. E Luciferase reporter scheme and graph showing luciferase activity (mean ± SD) under indicated conditions. (n = 4 per condition, p value by 1-way ANOVA). F Kinomic inhibition profile of CT-179. The protein kinases inhibited by CT-179 on the KINOMEscan array are marked as red nodes on the dendrogram of the human kinome. Node size indicates the levels inhibition of the kinases. G Phenotypic profiling of CT-179 with BioMAP Diversity PLUS panel. The profile plot shows the effects of CT-179 at 160 nM (orange), 630 nM (yellow) and 2.5 µM (green) on a panel of 12 primary cell systems (BioMAP 1-12) with 148 clinically-relevant biomarker readouts. H Plasma CT-179 concentrations over 24 hours after single CT-179 dose (left, PO right, IV). I Plasma and brain concentrations of CT-179 (left panel) and brain:plasma ratio (right) after 1 and 4 hours of single oral CT-179 administration (20 mg/kg). J Plasma CT-179 concentrations under indicated conditions. A, C, D Box plots show means ± SD, p value by 1-way ANOVA. E, I dots indicate individual biological replicates and columns indicate means. H–J n = 3 animals. Source data are provided as Supplementary Data 4 Source Data file.
