This study reports for first time the antioxidant capacity and cholinesterase inhibitory activity of the methanol extract of the lichen Vulpicida pinastri (Scop.) J.E. Mattsson & M.J. Lai and its chemical composition. This lichen specie with a greenish yellow foliose thallus was collected in Puerto Alto del Peñon, Zamora (Spain). Antioxidant capacity was assessed by in vitro tests (DPPH, ORAC and FRAP), total phenolic content by Folin-Ciocalteu method, cholinesterase inhibitory activity by Ellman’s colorimetric method and chemical composition by HPLC-UV method. The results showed that the values for antioxidant capacity were IC₅₀ 283.7 ± 31.7 microg/ml for DPPH, 1.5 ± 0.1 micromol TE/mg dry extract for ORAC and 25.4 ± 2.3 micromol of Fe²⁺ eq/g sample for FRAP. Moreover, total phenolic content had a value of 48, 9 ±4.8 microg GA/mg. Furthermore, IC₅₀ values were 0.19 ± 0.003 mg/mL for acetylcholinesterase inhibitory activity and 0.89 ± 0.018 mg/mL for butyrylcholinesterase inhibitory activity. Finally, the analysis of chemical composition revealed that the major secondary metabolites were vulpinic acid (9.3 ± 0.8), pinastric acid (41.9 ± 1.07) and usnic acid (36.5 ± 3.62). In conclusion, Vulpicida pinastri is a promising agent to further study for the prevention and treatment of Alzheimer’s disease based on its antioxidant and cholinesterase inhibitory activities.