Mps1–Targeted Molecular Design of Melatonin for Broad-Spectrum Antifungal Agent Discovery

Melatonin, a multifunctional class of natural products, has demonstrated antifungal activity, making it a promising candidate for developing antifungal agents. The mitogen-activated protein kinase (Mps1) within fungal pathogens has a target inhibitory effect of melatonin in fungi. We use a virtual screening strategy to design melatonin derivatives based on the melatonin-Mps1 targeting model. Of these, a multiflorane-substitution compound M-12 emerges as a potent antifungal agent, exhibiting broad-spectrum efficacy against eight phytopathogenic fungal species, and effectively reduces the severity of tomato gray mold, Fusarium head blight in wheat, Sclerotinia stem rot in rape, and peach brown rot. M-12 half-maximal effective concentration values (5.50 μM against Botrytis cinerea, 5.21 μM against Fusarium graminearum, 10.6 μM against Rhizoctonia solani, and 9.02 μM against Sclerotinia sclerotiorum) are better than those of commercial broad-spectrum fungicide azoxystrobin (55.0, 23.2, 46.5, and 17.7 μM, respectively). Antifungal activity of enantiomer (S)-M-12 (5.02 μM) is significantly greater than its (R)-enantiomer (23.6 μM) against B. cinerea. Molecular docking and transcriptome analysis reveal that M-12 achieves its antifungal effects by inhibiting Mps1 kinase, thereby suppressing fungal growth and virulence.