The aim of this work was to investigate the  phytochemical and anti-proliferative activity in vitro of the aerial parts of lotus peregrinus L. The preliminary phytochemical analysis of the aerial parts of lotus peregrinus L. (Family; Fabacea) revealed the presence of phenols, flavonoids, terpenoids, steroids, tannins and carbohydrates. Protein amino acids analysis showed the presence of 17 amino acids as protein amino acids. Aspartic acid (8.526 mg/g) and Glutamic acid (6.069 mg/g) represented as the major components of protein amino acids, respectively. Combined and free sugars analysis showed the presence of 12 free sugars. L-Rhaminose (21.9136) and Glucuronic (21.656 %) represented as the major free sugars, meanwhile the Xylose (29.425%); followed by Glucuronic (27.899%) represented as the major combined sugars. Twelve fatty acids were estimated; the highest percentage was that of Linoleic acid (18.489). Fourteen known hydrocarbon were found, the highest percentage was that of Eicosane (8.027%) in addition to three sterols; the highest percentage was that of Stigmasterol. two triterpene: alpha-Amyrin and lupeol in addition to one acyclic diterpene; Neophytadiene were detected. Anti-proliferative activity was carried out on the successive extracts (diethyl-ether, chloroform, ethyl acetate and ethanol 70% extracts) of lotus peregrinus L aerial parts to evaluate the Anti-proliferative properties. Diethyl ether was found to be potential anti-proliferative extract and promising natural agent. The chemical composition of the diethyl ethyl extracts can be useful in the chemosystematics of this species. Further studies will be needed to clarify the exact mechanism of Lotus peregrinus L. most active extracts as anti-cancer agent.