Leaves of Mouriri guianensis Aubl. (Melastomataceae), collected in the municipalities of Parnaíba and Campo Maior, Piauí, Brazil, were macerated with ethanol, resulting in ethanolic extracts of the leaves collected in Parnaíba (EEFP) and Campo Maior (EEFCM). Liquid-liquid extraction of these extracts produced hexane fractions from Parnaíba (FHP) and Campo Maior (FHCM). Phytochemical characterization of the fractions was performed using gas chromatography-mass spectrometry (GC-MS), allowing the recording of 39 compounds, with a predominance of isoprenoids, esters and fatty acids. The major compounds were β-sitosterol and phytol, with relative abundances of 26.24% and 16.14% in FHP, and 34.56% and 14.51% in FHCM, respectively. The fractions were evaluated against human tumor cell lines, including colorectal adenocarcinoma (HCT-116) and breast cancer (MDA-MB-231), as well as a non-tumoral murine fibroblast cell line (L929). The inhibition percentages were below 50%, indicating low toxicity. EEFP and EEFCM extracts exhibited inhibitory activity against acetylcholinesterase, with IC₅₀ values of 1.030 and 1.067 mg/mL, while the FHP and FHCM fractions showed IC₅₀ values of 26.17 mg/mL and 45.21 mg/mL, respectively. This study contributes to the phytochemical and biological knowledge of M. guianensis, highlighting its anticholinesterase e potential and low toxicity profile.