Background: Gymnema sylvestre also known as __ampersandsignlsquo;GURMAR__ampersandsignrsquo; is one of the important herbs in the Ayurvedic system of medicine and Indian proprietary medicines. Objective: In the present investigation, phytochemical screening, free radical scavenging, anti-inflammatory and anti-cancer activities were evaluated in vitro in the aqueous extract from the leaves of Gymnema sylvestre (AGS). Methods: Leaf samples were subjected with four different solvents (ethanol, acetone, ethyl acetate and distilled water) and the four extracts were screened for phytochemicals using different biochemical tests. AGS was subjected to fourier-transform infrared (FT-IR) spectroscopy to analyze its functional groups. Antioxidant activity of AGS was determined by measuring nitric oxide (NO) radical scavenging activity. The anti-inflammatory activity was studied using the inhibition of protein denaturation assay and anti-cancer activity was evaluated using MTT assay against MG63 cell lines. Results: Phytochemical screening confirmed the presence of phytoconstituents like alkaloids, carbohydrates, triterpenoids, proteins, phenols and flavonoids. The FT-IR spectrum with different peak values revealed the presence of various functional compounds in AGS and it showed significant NO radical scavenging activity with the EC50 value of 401.66 __ampersandsignmu;g/mL. AGS displayed strong anti-inflammatory activity (IC50 = 17.5 __ampersandsignmu;g/mL). Moreover, AGS also exhibited significant dose-dependent anti-cancer activity against MG63 cell line and the IC50 value was 19.5 __ampersandsignmu;g/mL. Conclusion: The results indicate that Gymnema sylvestre can act as a promising free radical scavenger, anti-inflammatory and anticancer agent against MG63 human osteosarcoma cell lines. Hence, G. sylvestre can be used as natural antioxidants for preventing the oxidative stress related degenerative diseases..