Prunus lusitanicaL. is a shrub belonging to the genusPrunusL. (Rosaceae family) that produces small fruits with none known application. Thus, the aim of this study was to determine the phenolic profile and some health-promoting activities of hydroethanolic (HE) extracts obtained fromP. lusitanicafruits, harvested from three different locations. Qualitative and quantitative analysis of extracts was performed using HPLC/DAD-ESI-MS and antioxidant activity was assessed by in vitro methods. Antiproliferative/cytotoxic activity was determined on Caco-2, HepG2, and RAW 264.7 cells, anti-inflammatory activity was assessed using lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, and the antidiabetic, antiaging, and neurobiological action of extracts was determined in vitro by assessing their inhibitory effect against the activity of α-amylase, α-glucosidase, elastase, tyrosinase, and acetylcholinesterase (AChE). Results showed thatP. lusitanicafruit HE extracts from the three different locations showed identical phytochemical profile and bioactivities, although small differences were observed regarding the quantities of some compounds. Extracts ofP. lusitanicafruits contain high levels in total phenolic compounds, namely, hydroxycinnamic acids, as well as flavan-3-ols and anthocyanins, primarily cyanidin-3-(6-trans-p-coumaroyl)glucoside.P. lusitanicafruit extracts have a low cytotoxic/antiproliferative effect, with the lowest IC50value obtained in HepG2 cells (352.6 ± 10.0 μg/mL, at 48 h exposure), but high anti-inflammatory activity (50–60% NO release inhibition, at 100 μg/mL extract) and neuroprotective potential (35–39% AChE inhibition, at 1 mg/mL), and moderate antiaging (9–15% tyrosinase inhibition, at 1 mg/mL) and antidiabetic (9–15% α-glucosidase inhibition, at 1 mg/mL) effects. The bioactive molecules present in the fruits ofP. lusitanicadeserve to be further explored for the development of new drugs of interest to the pharmaceutical and cosmetic industry.