Figure 10.

Dose-dependent CCR5 antagonism by the bifunctional chimera and the parent small-molecule inhibitors. (Top) Dose−response curves of the extent of the inhibition of RANTES (10 nM) stimulation of calcium mobilization measured for LJC240-containing compounds. Maraviroc was included as a control. (Bottom) Chemotaxis inhibition of CEM CCR5 cells to RANTES by LJC240−L4−UM15. One million CEM CCR5 cells were seeded in the apical chambers of 8 μm transwell inserts, and chemoattraction was induced using RANTES (10 ng/mL) in the presence or absence (control, No Chemokine) of the inhibitors maraviroc (MRVC); LJC240; LJC240−L4; or the peptide triazole chimera, LJC240−L4−UM15, at the IC₅₀ value for each compound. Data are presented as cells recovered in the basolateral chamber postincubation normalized to cells migrating to RANTES in the absence of inhibitor. Two-way ANOVA was used to compare the inhibitor-treated groups to the RANTES-only control group, and Student’s t-test was used to compare individual groups. Error bars represent means ± SEM; *p ≤ 0.05, **p ≤ 0.01. These data are representative of two independent experiments.