Various families of mushrooms contain important phytochemicals with significant potential. This study aimed to investigate the phytochemical prospecting, presence of psilocybin, antioxidant activities, and acetylcholinesterase inhibition in aqueous extracts of Cyathus striatus, Laternea dringii, and Marasmius haematocephalus. Aqueous extracts were produced from mushrooms, and phytochemical groups were determined. The total phenolic and flavonoid content, DPPH reduction capacity, and FRAP were quantitatively determined. The acetylcholinesterase inhibition assay was performed, and the results were expressed as AChE inhibition percentages. Phytochemical groups such as flavonoids, phenolics, alkaloids, organic acids, and aliphatic compounds were positively detected. For phenolics, the extracts showed values of 208.44, 134.11, and 100.09 mg GAE g TPC-1; for flavonoids, values of 45.12, 56.06, and 39.71 mg QE g TFC-1. The FRAP reduction capacity showed values of 7.56, 14.43, and 4.15 µM TE g-1, while for DPPH, the values were 100.07, 88.12, and 133.65 µg mL-1. Low, medium, and strong AChE inhibition activity was observed with values of 43.11%, 68.53%, and 77.14%, respectively, for C. striatus, L. dringii, and M. haematocephalus. The aqueous extracts of Cyathus striatus, Laternea dringii, and Marasmius haematocephalus exhibited various phytomolecules groups with potential biological activities observed in this study.