Nematodes parasitize ∼1/3 of humans world-wide. Although current anthelmintics are effective against many nematode infections, there is growing concern that resistance could threaten this mode of treatment. Pharmacological compounds that target xenobiotic detoxification pathways, which lead to multidrug resistance can greatly increase the useful life of current and future anthelmintics. The transcription factor SKN-1 is essential for the development of nematode embryos and activates the expression of ∼65 genes predicted to promote modification, conjugation, or export of drugs [1]. We have identified a major pathway regulating SKN-1 that is distinct from the major pathway regulating xenobiotic detoxification in mammals. SKN-1 also binds to target DNA by a unique monomeric mechanism relative to all other known basic leucine zipper factors [2,3]. Hence, SKN-1 is a promising target for the development of inhibitors that disrupt embryonic development, decrease stress resistance, and inhibit xenobiotic detoxification in nematodes without affecting homologous pathways in humans. We have successfully carried out a HTS screening of the MLSMR library to identify inhibitors of the transcriptional activation of the SKN-1 pathway in C. elegans through use of promoters of the SKN-1 target gene gst-4 (Pgst-4::GFP), which can be detected through simple fluorescence measurements. We report for the first time a small molecule inhibitor of the SKN-1 pathway, ML358, which resulted from a vanillamine derived scaffold identified from the HTS program and advanced through analog synthesis and structure-activity studies. ML358 is a potent (IC50 = 0.24□μM, Emax = 100%) and selective inhibitor of the SKN-1 pathway in C. elegans. ML358 is non-toxic to both C. elegans (LC50 > 64 μM) and Fa2N-4 immortalized human hepatocytes (LC50 > 50μM). In addition, ML358, has good solubility, permeability, chemical and metabolic stability in human and mouse liver microsomes. ML358 is a valuable probe to study the function and relevance of the transcription factor SKN-1 in drug detoxification and resistance in parasitic nematodes. Currently available drugs target only a specific enzyme or enzyme class and/or affect homologous human targets. By selective targeting of the SKN-1 pathway present in diverse nematode clades, ML358 can potentially lead to drug candidates that could be used as adjuvants to increase the efficacy of current anthelmintics.