Acrocarpus is a genus of flowering plants in the legume family Fabaceae which considered as a large and economically important family. This study aimed to carry out the biological activity screening on the total ethanol and successive extracts of Acrocarpus fraxinofolius (A. fraxinofolius) bark, for the first time. The biological activity studied embraced, management of diabetes in alloxan induced diabetic rats, cytotoxic activity against four human tumor cell lines and hepatoprotective activity against CCl4-induced hepatotoxicity in rats and the activity was studied by assaying the serum marker enzymes like AST, ALT, and ALP. Concerning this, the petroleum ether extract (PEE) showed the most bioactive extract where, the anti-diabetic activity exhibited by 100mg of PEE extract was 74.38% relative to metformin. It also showed a significant anti-proliferative activity against MCF-7 (IC50=2.35µg), Hela (IC50=3.85µg) and HEPG-2 (IC50=9.54µg) compared with Doxorubicin as reference drug. The hepatoprotective activity of the PEE was evidenced by a significant decrease in the liver function enzymes, i.e. AST, ALT and ALP by 29.18%, 28.26%, and 34.11%, respectively, using silymarin as the reference drug, compared to their concentration levels in an untreated group with liver damage induced by CCl₄. Based on the above outcomes, further phytochemical investigation including GC/MS analysis of its fractions, GLC analysis of its sterol fraction, column chromatography and TLC fractionation of PEE to separate its bioactive compounds were conducted.