In the current investigation, total phenolics and flavonoids of the methanolic extract obtained from the trunk bark of Acacia cyanophylla Lindl. were quantified by LC-HRMS technique. DPPH and ABTS reagents were employed to assay the antioxidant potential. The anti-tyrosinase and anti-α-amylase potentials were also assayed. The findings revealed that thirteen polyphenolic compounds were detected in the methanolic extract with trans-taxifolin (23.2 g/kg), as the major constituent. A. cyanophylla extract displayed a higher activity with DPPH test (IC50=10.14±1.00 μg/mL) than with ABTS (IC50=15.27±2.09 μg/mL). The same extract also exhibited interesting α-amylase inhibitory action (IC50 value of 4.00±0.17 μg/mL). Moreover, methanolic trunk bark extract exerted strong anti-tyrosinase capacity with an IC50 of 5.12±0.41 μg/mL in comparison to kojic acid (IC50=10.22±0.85 μg/mL) used as positive control. The antioxidant, anti-tyrosinase and anti-α-amylase potentials of the methanolic extract of A. cyanophylla trunk bark were reinforced by in silico molecular docking analyses, which confirmed the results of the in vitro tests.