This study investigates the pharmaceutical potential both in vitro and in silico of ethanolic propolis extract from three Algerian regions namely Tahir-Jijel (TAH), Oued Athmania-Mila (ATH) and Oued Zhor-Skikda (OZ). Twenty-three compounds were identified via a high-performance liquid chromatography-diode array detector, with key constituents including caffeic acid, cynarin, chrysin, naringin and hesperetin. Moreover, antioxidant and anti-Alzheimer activities were assessed by multiple assays. Antimicrobial activity was tested against four pathogens and one yeast. The TAH extract showed the highest antioxidant activity, and ATH exhibited the strongest anticholinesterase activity. Extracts demonstrated potent inhibition against Staphylococcus aureus and Micrococcus luteus ATCC 4698. Docking studies revealed that cynarin was the most potent acetylcholinesterase inhibitor, and quercetin was the most potent butyrylcholinesterase inhibitor. These compounds were found to bind to both the catalytic site and the peripheral site, showing greater inhibitory potency than galantamine.