Metabolomics-guided network pharmacology and molecular modeling suggest bioactive components of Chiliadenus montanus...

Chiliadenus montanus (Vahl.) Brullo (Asteraceae) is a traditional medicinal plant with reported anti-inflammatory properties; however, its potential anticancer activity and underlying molecular basis remain insufficiently characterized. In this study, a bioactivity-guided integrative approach combining metabolomics, biological evaluation, network pharmacology, and computational modeling was applied to explore the antiproliferative potential of the acetone extract of C. montanus (CME). CME exhibited cytotoxic activity against a panel of 14 human cancer cell lines (IC₅₀ = 1.41-24.78 µg/mL), with HepG2, Panc-1, and HCT-116 cells showing the highest sensitivity relative to a normal fibroblast control (selectivity index > 13). Flow cytometry analysis revealed substantial apoptosis induction and cell-cycle perturbation at the G₀/G₁ and S phases. Consistently, protein expression analysis showed upregulation of p53, Casp7, and BAX together with downregulation of BCL-2, resulting in an increased BAX/BCL-2 ratio across the responsive cell lines, supporting activation of mitochondrial apoptotic signaling. UPLC-TOF-MS/MS profiling tentatively identified 162 metabolites, predominantly flavonoids and organic acids, including luteolin, naringenin, kaempferol derivatives, and γ-linolenic acid. Network pharmacology identified 87 overlapping targets associated with liver cancer-related pathways, highlighting COX-1 (PTGS1), 5-LOX (ALOX5), and oxidative-stress-related proteins as central nodes. Consistent with these predictions, CME inhibited COX-1 and 5-LOX activity (in vitro) with IC₅₀ values of 7.25 µg/mL and 3.21 µg/mL, respectively, and displayed antioxidant activity in DPPH, ABTS, and FRAP assays. Moreover, CME reduced cellular levels of PGE₂, IL-6, and TNF-α in HepG2 cells, indicating suppression of inflammatory signaling. Molecular docking further suggested favorable interactions between several identified metabolites and inflammation-related targets. Overall, CME activity is associated with modulation of apoptotic and inflammatory signaling pathways, supported by biochemical, cellular, metabolomic, and computational analyses. While these findings provide mechanistic insights into the biological activity of C. montanus, further studies, including compound isolation and in vivo validation, are required to confirm these effects and their therapeutic relevance.