Alzheimer’s disease (AD) is a primary degenerative disease of the central nervous system. Increased level of the enzyme acetylcholinesterase (AChE) plays a key role in the hydrolysis of the neurotransmitter Acetylcholine which worsens the condition of cognitive dysfunction. Among the pathologic hypotheses of Alzheimer’s disease, cholinergic deficit and oxidative stress have been implicated as two major hallmarks. Hence, inhibition of cholinesterase and oxidation are the two important strategies in the development of a drug for AD. The in vitro antioxidant and anticholinesterase activities of ethanolic extracts of Nyctanthes arbortristis Linn. were evaluated using enzymatic and chemical methods. Antioxidant potentials were evaluated by the DPPH assay and lipid peroxidation method. Anticholinesterase activity was measured by the modified Ellman method. The molecular docking study was carried out using the molecular docking tool UCSF Chimera tool to explain the interaction of the major chemical constituents with the enzymes. In antioxidant in vitro evaluation, IC50 values of the ethanolic extract of Nyctanthes arbortristis were found to be 64.96 μg/ml and 79.27 μg/ml for DPPH and lipid peroxidation respectively; The ethanolic extract of Nyctanthes arbortristis showed potent anticholinesterase effects with an IC50 value of 28.54 μg/ml. The docking experiments showed that Quercetin has the highest binding score among the six major identified compounds. Quercetin also mimics the standard drug galantamine that shows the same H-bond interaction with THR62 amino acid residue resulting in more potent inhibitory activity than galantamine. The ethanolic extract of flowers of Nyctanthes arbortristis showed promising antioxidant and anticholinesterase activity and can be used to treat Alzheimer’s disease