Vitex altissima L, belonging to the Verbanaceae family, is available in India's Eastern Ghats and Deccan plateau. They were traditionally used to treat rheumatism. Six Iridoid glycosides and two Iridoids agunoside and negunoside with potent antioxidant potential were isolated, and Lignan altisonin, flavonoid 2-o-hydroxybenzoylorientin, were found in leaves. The compounds showed high binding energy with reverse transcriptor protein and exhibited antiviral action comparable with zidovudine, and stavudine. GC-MS analysis revealed the presence of n hexadecanoic acid, 12-octadecadienoic acid, and squalene. HPTLC fingerprinting analysis was developed to quantify and identify the marker compounds. Leaf methanolic extracts exhibited antibacterial activity against five strains of Gram-positive and Gram-negative bacteria. The sesquiterpenoid compounds were found to exhibit anticancerous and antioxidant action. The V.altissima was proved to have promising acts so could be further explored for their pharmacological actions and formulated.