Steroidal compounds from Daedaleopsis confragosa suppress breast cancer via Src inhibition.

Daedaleopsis confragosa, a medicinal mushroom, produces endoperoxide-containing steroids with anticancer activity, but their mechanisms remain unclear. This study investigated the anticancer potential of steroidal compounds from D. confragosa as food-derived bioactive compounds. Network pharmacology identified Src and PI3K-Akt pathway as key targets. Among seven isolated compounds, ergosterol peroxide (compound 1), 9,11-dehydroergosterol peroxide (compound 2), 5α,8α-epidioxyergosta-6-en-3-ol (compound 3), and (3β,5α,8α)-5,8-epidioxyergost-6-en-3-ol (compound 5) significantly inhibited MDA-MB-231 breast cancer cell viability, with compound 2 exhibiting the highest potency. These compounds induced cell death and reduced phosphorylated Src levels, with similar effects in 3D spheroid models. Molecular docking and dynamics simulations revealed that compound 2 and its metabolites (2-1,2-2) interact with Src near the ATP-binding site. Notably, metabolite 2-2 exhibited stronger binding affinity (-56.93 kcal/mol) than ATP (-40.29 kcal/mol). These findings suggest that steroidal compounds from D. confragosa inhibit Src kinase by blocking ATP binding, demonstrating significant therapeutic potential.