Integrated network pharmacology and in vitro/in silico evaluation of the bioactive potential of Gundelia komagenensis fırat extracts.

Abstract

Gundelia komagenensis Fırat, an endemic member of the Asteraceae family, was investigated for its phytochemical composition, antioxidant, enzyme inhibitory, cytotoxic, and computationally predicted pharmacological properties. Methanol (MeOH) and dichloromethane (DCM) extracts prepared from the aerial parts of Gundelia komagenensis were evaluated using ABTS and FRAP antioxidant assays, enzyme inhibition tests against acetylcholinesterase (AChE), α-glucosidase, and α-amylase, LC-MS/MS profiling, MTT-based cytotoxicity assays, and integrated in silico analyses. Both extracts exhibited antioxidant activity; among the extracts, DCM had a lower ABTS IC₅₀ value than MeOH (13.7 vs.14.5 µg/mL). DCM also had lower IC₅₀ values for AChE, α-glucosidase, and α-amylase than MeOH (1.132-1.372 µg/mL). LC-MS/MS revealed solvent-dependent profiles, with chlorogenic acid, vanillin, trans-cinnamic acid, and gentisic acid predominating in MeOH, and vanillin, trans-cinnamic acid, and luteolin in DCM. The MeOH extract of G. komagenensis did not exhibit significant cytotoxicity against either MIA PaCa-2 or HUVEC cells. In contrast, the DCM extract reduced MIA PaCa-2 cell viability in a concentration-dependent manner while exerting relatively limited effects on HUVEC cells, suggesting a selective cytotoxicity tendency under the tested conditions. Computational analyses identified chlorogenic acid as the most promising bioactive constituent. Molecular docking showed the highest binding affinities toward AChE, α-glucosidase, and α-amylase, while molecular dynamics simulations and MM-GBSA calculations supported the stability of the corresponding protein-ligand complexes. Network pharmacology suggested that the identified phenolics may regulate disease-related pathways through AP-1/NF-κB-associated targets. Chlorogenic acid emerged as a key multi-target candidate warranting further experimental validation.