Structure-guided discovery of 4-phenyl-5-p-tolyloxymethyl-4H-1,2,4-triazole thioethers as 15-lipoxygenase inhibitors using integrated in vitro, structure-activity relationship, in silico, cytotoxicity towards MCF-7 cell line, DNA docking and MD simulation methods.

15-Lipoxygenase (15-LOX) has emerged as a strategic target for therapeutic intervention in inflammatory diseases and the advancement of malignancy owing to its involvement in arachidonic acid metabolism. In this regard, in the pursuit of new 15-LOX inhibitors, a new series of alkyl/aralkyl derivatives (6a-h) of 4-phenyl-5-p-tolyloxymethyl-4H-1,2,4-triazole was designed and synthesized and then screened against the target enzyme. The synthesis started by the conversion of 2-p-tolyloxyacetic acid (a) into its corresponding ester (1), hydrazide (2), thiosemicarbazide (3) and 4-phenyl-5-p-tolyloxymethyl-4H-1,2,4-triazole-3-thiol (4). The target compounds 6a-h were achieved by the reaction of alkyl/aralkyl halides (5a-h) with 4 in the presence of alcoholic KOH. The intermediates and the products were characterized using advanced spectroscopic and spectrometric techniques. The screening data for compounds 6a-h showed them as potent inhibitors (IC from 0.53±0.11 to 3.67±0.28 μM). Structure-activity relationship (SAR) studies revealed that the compounds with greater lipophilic character demonstrated greater activity than those with less lipophilic assembly. The compounds were not toxic towards blood mononuclear cells. Compounds 6g displayed an IC value of 5.32 μM when tested against the MCF-7 breast cancer cell line, followed by 6f, 6d and 6b compared with that of quercetin. Pharmacodynamic studies revealed good druggability profiles. In silico studies supported the in vitro data. DNA docking studies revealed major groove binding. Docking studies of the analogues with the components of the NF-κB signalling pathway (ID:4KIK and 1VKX) exhibited excellent binding affinities, predicting a potential role in inhibiting the canonical pathway of gene regulation. Overall, this multifaceted study foresees 1,2,4-triazole thioethers as potential dual-action molecules and predicts targeting the NF-κB pathway components for interventions linked to chronic inflammation and cancer.