From the deep‐sea‐derived Fusarium sp. ZEN‐48, four known compounds were obtained. Their structures were established by extensive analyses of the NMR, HR‐ESI‐MS, and the X‐ray crystallographic data as brefeldin A (BFA, 1), brevianamide F (2), N‐acetyltryptamine (3), and (+)‐diaporthin (4). Although BFA was extensively investigated for its potent bioactivities, its role on TNFα‐induced necroptosis was incompletely understood. In this study, BFA showed significant inhibition on TNFα‐induced necroptosis by disrupting the necrosome formation and suppressing the phosphorylation of RIPK3 and MLKL (IC₅₀=0.5 μM). While, it had no effect on TNFα‐induced NF‐κB/MAPKs activation and apoptosis. The finding raised significant implications of BFA for necroptosis‐related inflammatory disease therapy and new drug development from marine fungi.