Fig. 10: JNJ-67869386 and JNJ-799760 bind to the same pocket.

a Kinetics of current recovery from inhibition by 100 nM JNJ-67869386, 1 µM JNJ-799760 or 100 nM JNJ-67869386 + 1 µM JNJ-799760. The steady-state current in the presence of compound is subtracted before normalization. Data are fitted to a single or double exponential function (dashed lines) with time constants of 9.1 ± 0.4 s (n = 11; solid circles), 3.1 ± 0.2 s/14.9 ± 1.6 s (n = 5; half-filled circles) and 3.9 ± 0.2 s (n = 5; open circles), respectively, for JNJ-67869386, JNJ-67869386 + JNJ-799760, and JNJ-799760. The three groups of data are significantly different from one another (p < 0.001; Two-way ANOVA). The holding pH is 8.2. b Docking of JNJ-67869386 (salmon carbons) to the binding site of JNJ-799760 on ΔASIC1 (green carbons). Stick representation of JNJ-67869386 and channel residues with which it interacts. Specific interactions between JNJ-67869386 and ΔASIC1 are shown as yellow dotted lines labeled with distances in Å. c Concentration-dependent inhibition of pH 6.0-induced currents of rASIC1a WT, F98A, and Y340A channels by JNJ-67869386 (solid symbols) and JNJ-799760 (open symbols). Data are fitted to a logistic function (dashed and dotted lines). For JNJ-67869386, IC₅₀ = 28.9 ± 3.6 nM (n = 7), 626.3 ± 11.0 nM (n = 4) and 571.8 ± 176.9 nM (n = 5), respectively. For JNJ-799760, IC₅₀ = 24.9 ± 1.3 nM (n = 4), 725.9 ± 356.1 nM (n = 4) and 260.0 ± 89.8 nM (n = 4), respectively. All mutant channel data are significantly different from the corresponding WT (p < 0.001; two-way ANOVA).