Extraction solvent shapes the phytochemistry, multi-target bioactivity, and machine learning-predicted anticancer targets of Seseli libanotis

Seseli libanotis (L.) W. D. J. Koch (Apiaceae) is a wild vegetable traditionally consumed in eastern Anatolia, but its non-volatile phytochemistry and multi-target bioactivity remain insufficiently characterized. This study evaluated how extraction polarity shapes the chemical and biological profiles of four aerial-part extracts. The extracts were analyzed for total phenolic and flavonoid contents, chemical composition, antioxidant activity, inhibition of five enzymes, cytotoxicity against three tumor cell lines and human embryonic kidney (HEK-293) cells, and in silico molecular docking with enzyme targets. The ethanol/water extract showed the highest total phenolic content. Qualitative liquid chromatography-mass spectrometry (LC-MS) profiling showed broad coverage of caffeoylquinic acids and flavonoid glycosides in the polar extracts. Antioxidant activity was solvent- and assay-dependent: water was most active in the 2,2-diphenyl-1-picrylhydrazy (DPPH), cupric reducing antioxidant capacity (CUPRAC), and ferric reducing antioxidant power (FRAP) assays, ethanol/water in the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay, and ethyl acetate (EA) in the phosphomolybdenum assay. Ethanol and ethanol/water showed the highest acetylcholinesterase (AChE) and α-glucosidase inhibition, without significant differences between them; EA was most active against butyrylcholinesterase (BChE) and α-amylase. Molecular docking analyses suggested favorable binding modes of selected phytochemicals within the active sites of the investigated enzyme targets. Subsequent molecular dynamics simulations validated these interactions by demonstrating stable binding conformations sustained through persistent hydrogen-bonding networks. The ethanol/water extract was the most cytotoxic against three tumor cell lines (IC₅₀ 31.77–41.25 µg mL⁻¹). Annexin V and propidium iodide staining (Annexin V/PI) and acridine orange/ethidium bromide (AO/EB) staining indicated apoptosis-associated viability loss; however, similar activity against HEK-293 cells indicated limited tumor selectivity. Extraction solvent was the principal determinant of phytochemical composition and bioactivity, while the low selectivity highlights the need for fractionation and compound-level validation.