The wide range of secondary metabolites in medicinal plant extracts are used to treat and manage various diseases worldwide but more recently plant-mediated bio-nanotechnology is rapidly generating great research interest. Herein, the in-vitro study of the antidiabetic, anti-inflammatory, antiglycation, and antioxidant potentials of silver nanoparticles (AgNPs) synthesized by Justicia secunda aqueous extracts was evaluated. The synthesized AgNPs was characterized by UV-visible spectrophotometer, Fourier transform infrared spectroscopy (FT-IR) and Scanning electron microscopy (SEM). The antioxidant activity was evaluated via reducing power, 2,2-diphenyl-1-picrylhydrazyl (DPPH), total antioxidant capacity (TAC) and nitric oxide scavenging assays; antidiabetic activity was evaluated with α-glucosidase and α-amylase inhibitory assays; antiglycation activity was evaluated using fructosamine inhibitory assay, and anti-inflammatory activity was performed using trypsin inhibitory and albumin denaturation assays. The AgNPs from all the plant parts showed good biological potentials investigated compared to the plant extracts however, the synthesized AgNPs using the leaf aqueous extract (AgNPs-JsL) showed a better antioxidant, antidiabetic, antiglycation, and anti-inflammatory potentials with an essential quality absorption band in the ultraviolet region of 410 nm and average size distribution of 50.47nm. In conclusion, all the parts of J. secunda showed substantial antidiabetic, antioxidant, antiglycation and anti-inflammatory potentials with high yield of various bioactive metabolites, however, the AgNPs-JsL was more potent followed by the flower, stem and roots extracts, respectively.