Integrative network pharmacology and experimental evaluation reveal TNBC-related biological activity and toxicity of Euphorbia umbellata crude latex.

Abstract

Euphorbia umbellata latex is used in Brazilian traditional medicine, including empirically for cancer treatment. However, its chemical composition, molecular targets, pharmacological properties, and biological effects in triple-negative breast cancer (TNBC) remain insufficiently characterized. This study investigated the TNBC-related biological activity and toxicity of E. umbellata crude latex by integrating computational analyses with experimental evaluation in a 4T1 cell model. Eighteen compounds previously identified in E. umbellata crude latex were curated from phytochemical studies and evaluated for predicted pharmacokinetic and toxicological properties. Compound-associated human targets were integrated with TNBC-related genes, followed by protein-protein interaction network construction, functional enrichment analysis, hub-gene identification, and molecular docking. The effects of crude latex on cell viability, post-treatment recovery, and wound closure were evaluated in 4T1 murine TNBC cells, while acute toxicity was assessed using the Artemia salina lethality assay. The latex-derived compounds exhibited heterogeneous ADMET profiles, with generally low predicted mutagenic, carcinogenic, and hepatotoxic potential. A total of 357 compound-associated targets were identified, of which 204 overlapped with TNBC-related genes. Enrichment analyses highlighted pathways associated with apoptosis, inflammatory responses, oxidative stress, and immune checkpoint regulation. Molecular docking indicated favorable predicted interactions with EGFR, AKT1, and PARP1, whereas PTGS2 showed a more restricted interaction profile. Experimentally, crude latex markedly reduced 4T1 cell viability, with an estimated IC50 interval of 2.15-2.76 mg/mL, and impaired wound closure during acute exposure. However, both effects were partially reversed after treatment removal. The latex also induced concentration-dependent lethality in A. salina, with an estimated LC50 of 3.25 mg/mL. Together, these findings indicate that E. umbellata latex represents a chemically diverse source of bioactive compounds capable of interacting with TNBC-associated molecular networks, while also showing biological activity in a 4T1 murine TNBC model. Nevertheless, the partial recovery of 4T1 cells after treatment removal and the toxicity observed in A. salina indicate that crude latex should not be considered directly suitable for therapeutic use and that further fractionation, chemical standardization, mechanistic validation, and safety studies are required.