The aim of the present study was to determine the antimicrobial, anti-inflammatory, antioxidant and cytotoxic activities of Hibiscus rosa sinensis and Vitex negundo. Hibiscus rosa-sinensis Linn. (Family Malvaceae) leaves had been used in ethnomedicine to treat a variety of human diseases including aphrodisiac, hypertension, wound healing, diabetes mellitus and cancer. In traditional medicine, the leaves of Hibiscus rosa-sinensis were used to treat dysentery and diarrhoea, to promote the draining of abscesses and as an analgesic. Natural products contained high levels of bioactive compounds that targeted a complex network of proteins involved in a variety of diseases. Vitex negundo (V. negundo), also known as the "chaste tree," was an ethnobotanical significant plant with powerful medicinal properties. In addition to its anti-diabetic properties, V. negundo leaf extract had a beneficial effect on glycoprotein metabolism. Antimicrobial activity was assessed using the agar well diffusion method and MIC assay. Antioxidant activity was evaluated H2O2 assay, FRAP assay and DPPH assay. Anti-inflammatory activity was tested using egg albumin denaturation and membrane stabilisation methods. All activities demonstrated very good results for the herbal formulation. Natural products had a high propensity to target specific proteins while causing fewer side effects and lower toxicity and greater effectiveness. Aqueous extract of V. negundo and Hibiscus rosa sinensis were used as treatment for a variety of diseases and their products have become commercialised, increasing their demand. Overall, the formulation contributed to health protection and showed potential in the management of autoimmune diseases.