Integrated In Silico-to-In Vitro Validation of a Dual-Oil Phytochemical- Formulation Against Malignant Melanoma Cell Line; Multi-Target Docking, Polypharmacology, and Neutral Red Uptake (NRU)-Based Cytotoxicity Profiling Versus Cisplatin

This study integrates a multi-target in silico screening campaign with in vitro experimental validation to assess a dual-oil phytochemical formulation (cold-pressed Prunus dulcis oil combined with Pinus sylvestris essential oil enriched in α-pinene; commercially referred to as “Naevus Support”) as a candidate adju-vant/alternative strategy against malignant melanoma. First, a comparative molecular docking workflow was applied across a melanoma-relevant target panel spanning the MAPK axis (BRAF, MEK1, ERK2), cell-cycle control (CDK4/6), DNA damage signaling (PARP1), inflammatory lipid signaling (COX-2), and melanogenesis-associated enzymes (tyrosinase), benchmarking major oil constituents and derived chemo-types against standard-of-care inhibitors. Docking energetics and pose-level interaction forensics supported a polypharmacology profile consistent with concurrent suppression of oncogenic signaling nodes and mi-croenvironmental permissive pathways. Second, the same formulation was tested in a Neutral Red Uptake (NRU) viability assay on B16F10 malignant melanoma cells and MRC-5 human fibroblasts, using cisplatin as a reference cytotoxic agent. Across a concentration range of 3–0.045% (v/v) for oils and 20–0.18 mM for cisplatin, the dual-oil formulation induced a dose-dependent reduction of melanoma viability while main-taining comparatively lower toxicity on fibroblasts, indicating a therapeutically relevant selectivity window. Individual-oil profiling suggested that the combined formulation’s anticancer activity cannot be explained by single-oil effects alone, supporting a true inter-oil synergistic enhancement that aligns with the mul-ti-node in silico predictions. Collectively, these data provide a coherent in silico-to-in vitro rationale for further mechanistic follow-up (target deconvolution, pathway readouts, and lipidomic/ROS endpoints) and in vivo translation.