Global challenges with anthelmintic failure and resistance development lends impetus to the development of new nematocides (anthelmintics) with novel mechanism(s) of action. The free-living nematode Caenorhabditis elegans is as an important model organism used for drug discovery and a powerful tool for anthelmintic screening, evaluation and target deconvolution. Previously, we conducted a whole-organism phenotypic screen of the ‘Pandemic Response Box’ (from Medicines for Malaria Venture, MMV) and identified a hit compound, called ABX464, with activity against C. elegans. Here, we explored this nematocidal pharmacophore on C. elegans, and then tested a series of 46 analogues for human hepatoma (HepG2) toxicity, revealing five compounds whose potency was similar or greater than that of ABX464. Subsequently, we employed thermal proteome profiling (TPP), protein structure prediction and an in silico docking algorithm to identify prime ABX464-target candidates. Taken together, the findings from this study contribute significantly to the early-stage drug discovery of a new nematocide based on ABX464. Future work is aimed at validating the ABX464-protein interactions identified here, and at assessing of ABX464 and associated analogues against a panel of parasitic nematodes, towards the development of a new anthelmintic with a novel mechanism of action.