Fig. 4: Nemacol analogs demonstrate nematode-selective activity.
a Nemacol-1 (black) and (-)Vesamicol (red) competitive displacement of [3H] Vesamicol binding of rat VAChT. Shown is the binding curve for each of two biological replicates (N = 2) tested in technical triplicate (n = 3) reporting the SEM for each binding curve. The P value comparison of the two sets of curves was calculated using Fisher’s Method (P < 10−15 for both; see 'Methods'). The determined Ki and asymmetrical confidence interval for (−)Vesamicol was 19.5 nM (17.7 nM–21.6 nM) and 22.4 nM (17.2 nM−29.4 nM) for each biological replicate. The determined Ki and confidence interval for nemacol-1 was 875 nM (665 nM–5.99 µM) and 661 nM (503 nM–868 nM) for each biological replicate. b Comparison of Vesamicol and Nemacol analog in vivo potency and rat VAChT binding affinity. Details of the means and confidence intervals are in (c) and Supplementary Table 1. c Structure activity relationship summary of Nemacol analog activity across nematode species and rat VAChT inhibition constants (Ki). C. elegans and Pristionchus pacificus EC50s are derived from biological triplicates (N = 3) with technical duplicate (n = 2) scoring 18 animals per condition. See Supplementary Figs. 4 and 5 for the dose-response curves. Dirofilaria immitis microfilariae immobility EC50 data are from triplicate (N = 3) measurement of ~250 animals per condition after 72 h of drug exposure. D. immitis L3/L4 immobility EC50 data are derived from singlet observations from 20 freshly isolated L3s after 72 h of incubation (see 'Methods'). NA no activity. The inhibition constant (Ki) confidence intervals for interaction with rat VAChT are presented in Supplementary Table 1. The grey cells indicate that the experiment was not done.
