Euphrasia officinalis L., commonly known as eyebright, is a medicinal plant used in folk medicine for eye disorders and memory loss. Due to its abundance of compounds with proven neuroprotective properties, there has been growing interest in exploring eyebright’s potential health benefits, particularly for preventing or treating neurodegenerative diseases like Alzheimer’s disease. Here, seven distinct extracts were generated using solvents of different polarities, consecutively, from plants grown in Greece. The extracts were chemically characterized and assessed for their antioxidant, anticholinesterase, and anti-neurotoxic potentials. Our findings demonstrated eyebright’s notable antioxidant capacity with five extracts exhibiting significant anti-neurotoxic properties by enhancing cell viability by 17.5 to 22.6% in human neuroblastoma cells exposed to neurotoxic amyloid-beta peptides. The ethyl acetate and butanolic extracts were the most effective across all assays, likely due to their high concentrations of active compounds. Therefore, eyebright may be harnessed for developing functional foods, supplements, and pharmaceuticals with potential benefits against Alzheimer’s disease. This study marks the first identification of neuroprotective properties in a Euphrasia species, highlighting its broader therapeutic potential and paving the way for further research.