Discovery of ML358, a Selective Small Molecule Inhibitor of the SKN‑1 Pathway Involved in Drug Detoxification and Resistance in Nematodes

Nematodes parasitize ∼1/3 of humans worldwide, and effective treatment via administration of anthelmintics is threatened by growing resistance to current therapies. The nematode transcription factor SKN-1 is essential for development of embryos and upregulates the expression of genes that result in modification, conjugation, and export of xenobiotics, which can promote resistance. Distinct differences in regulation and DNA binding relative to mammalian Nrf2 make SKN-1 a promising and selective target for the development of anthelmintics with a novel mode of action that targets stress resistance and drug detoxification. We report <b>17</b> (<b>ML358</b>), a first in class small molecule inhibitor of the SKN-1 pathway. Compound <b>17</b> resulted from a vanillamine-derived hit identified by high throughput screening that was advanced through analog synthesis and structure–activity studies. Compound <b>17</b> is a potent (IC₅₀ = 0.24 μM, <i>E</i>_max = 100%) and selective inhibitor of the SKN-1 pathway and sensitizes the model nematode <i>C. elegans</i> to oxidants and anthelmintics. Compound <b>17</b> is inactive against Nrf2, the homologous mammalian detoxification pathway, and is not toxic to <i>C. elegans</i> (LC₅₀ > 64 μM) and Fa2N-4 immortalized human hepatocytes (LC₅₀ > 5.0 μM). In addition, <b>17</b> exhibits good solubility, permeability, and chemical and metabolic stability in human and mouse liver microsomes. Therefore, <b>17</b> is a valuable probe to study regulation and function of SKN-1 <i>in vivo</i>. By selective targeting of the SKN-1 pathway, <b>17</b> could potentially lead to drug candidates that may be used as adjuvants to increase the efficacy and useful life of current anthelmintics