This research aims to study the biogenic synthesis and characterization of Hypecoum pendulum extract-induced silver nanoparticles (HP-AgNPs) and to investigate its antioxidant, anti-alpha-amylase, and anti-acetylcholinesterase (AChE) activities. UV spectrometry at 417 nm with an surface plasmon resonance (SPR) peak of 1.54 confirmed the biosynthesis of HP-AgNPs. X-ray diffraction (XRD) analysis demonstrated that HP-AgNPs are crystalline with a 36 nm size. The average size of 80 nm was confirmed by scanning electron microscopy (SEM) analysis. The functional moieties bearing compounds accountable for the encapsulation and stability of HP-AgNPs were assessed by the Fourier transform infrared (FTIR) technique. Energy dispersive X-ray (EDX) demonstrated that silver was the main element (49%) of HP-AgNPs. HP-AgNPs showed the promising antioxidant potential of ABTS (71.57%), diphenyl-1-picrylhydrazyl (DPPH, 69.46%), H 2 O 2 (66.94%), ferric chloride (73.71%), and ammonium molybdenum (65.7%). HP-AgNPs exhibited significant enzymatic inhibition potential with IC 50 of 48.8 ± 4.03 μg/mL (α-amylase) and 51.78 ± 3.16 μg/mL (AChE). Kinetics studies showed that HP-AgNPs produced mixed-type inhibition against α-amylase (K m increased from 40.65 to 131.44%, whereas V max decreased from 4.14 to 9.77%) and non-competitivetype inhibition (V max decreased from 17.05 to 62.03% and K m remain constant) against AChE. Finally, it may be concluded that HP-AgNPs may possess anti-Alzheimer, anti-diabetic, and antioxidant capabilities.