Background: The study of free radical chemistry has received a considerable lot of attention recently. Our bodies produce reactive oxygen and nitrogen species and free radicals as a result of a variety of endogenous processes, exposure to various physiochemical situations, or pathological conditions. For optimum physiological function, free radicals and antioxidants must coexist in balance. Oxidative stress results when the body's defences against free radicals are overpowered. As a result, free radicals damage lipids, proteins, and DNA and cause a variety of human disorders. Therefore, using antioxidants from an external source can help to manage this oxidative damage. In contrast to synthetic antioxidants, which are either added to food to increase its shelf-life or are synthesised by plants and found in the foods we consume, natural antioxidants are produced by plants (for example, vitamins and other naturally-occurring substances in our food) (e.g. BHT). Annona squamosa. (Annonaceae, Family). Hindi-speaking locals refer to it as "Sitaphal." Insecticidal, purgative, laxative, astringent, anti-inflammatory, antidiabetic, anti-ulcer, anti-oxidant, antimalarial, and antibacterial are only a few of the pharmacological effects of the plant. Method: In current study HO-1 protein selected as target protein. The Auto Dock software used a grid-based docking algorithm to determine the bond. Using the Merck Molecular Force Field, 2D structures of compounds were created, transformed to 3D, and then energetically decreased up to an arms gradient of 0.01. (MMFF). Result: Flavonoids of A.squamosa found to be effective antioxidant component and effectively binds to be target protein HO-1 with binding energy -6.18 & -5.26 kcalmol-1for quercetin and rutin respectively. Conclusion: The finding of the in-silico molecular docking showed that both lead compound is effective binds & inhibitory action on target protein. The molecular docking of ligands like quercetin and rutin with human HO-1 receptor revealed that it has exhibited the chemical interaction with the amino acids in the active pockets.