Medicinal plants have therapeutic potential and are used worldwide to treat various diseases. Vitex negundo Linn.; family Lamiaceae has been used for the treatment of Internally alternative, aromatic, bitter and vermifuge anodyne, tonic, febrifuge, expectorant and diuretic, skin diseases, pruritus, helminthiasis, rheumatism and puerperal fever. This study was designed to investigate the phytochemical and anti-inflammatory activity of methanolic extract of Vitex negundo Linn. in parasitized and unparasitized form (VNMpar and VNMunpar). The anti-inflammatory activity of methanolic extract of Vitex negundo Linn. in parasitized and unparasitized form was evaluated by inflammatory models of carrageenan induced paw edema in rats and acetic acid induced vascular permeability in Mice. The result that preliminary phytochemical analysis showed presence of flavonoids, triterpenoids, tannins, phenolic compounds along with primary metabolites. Methanolic extract of Vitex negundo Linn. in parasitized form significantly decreased carrageenan-induced paw edema and acetic acid-induced capillary permeability. Paw edema is dramatically reduced at 3, 4, and 5 h after carrageenan injection by VNMpar at 400 mg/kg along with vascular permeability too. In conclusion, the findings suggested that methanolic extract of Vitex negundo Linn. in parasitized form produced potential anti-inflammatory activity than unparasitized form which support the claim for its traditional use in the treatment of various diseases.