Avocado is an oil-rich fruit pleasant organoleptic characteristic, high nutritional value and health benefits due to its content of bioactive molecules. The objective of the current study was assessment of the antioxidant, anti-cancer and anti-arthritic activities of acetogenin-rich extract of avocado pulp. Total phenolic, total flavonoids, fatty acids profile, phytosterols and hydrocarbons of the extract were determined. Acetogenin was detected using GC-MS. The antioxidant activity of the extract was determined using DPPH, ABTS and FRAP assay. The cytotoxicity of the extract was assessed in different cancer cell lines (liver, colorectal and breast carcinoma cells). The anti-arthritic activity of the extract was evaluated in adjuvant induced arthritis (AIA) model in rats. The results revealed that acetogenin-rich extract of avocado contains phenolic compound (58.6±0.579 mg GAE/g extract) and total flavonoids (10.8±0.449 mg QE/g extract). GC-MS analysis revealed the presence of persediene as one of acetogenin compounds by 1.1% in the extract. The extract possessed antioxidant activity in all the studied methods and the maximum activity was against ABTS free radical. The IC50 of the extract against hepatocellular carcinoma cells (HEPG2), breast adenocarcinoma cells (MCF7) and colorectal cancer cells (HT29) was 8.1, 52.1 and 11.3 µg/ml, respectively. The extract showed significant anti-arthritic activity in rats through reduction of inflammation volume, improvement in the antioxidant status of arthritic rats as observed by reduction of plasma levels of malondiadehyde and elevation of the antioxidant enzymes (SOD, catalase and glutathione peroxidase) in association with reduction of the inflammatory marker TNF-α. The extract showed significant improvement in the histopatological studies of the joint tissue of arthritic rats. In conclusion: Acetogenin-rich extract of avocado pulp can be used as alternative treatment for prevention or treatment of cancer and rheumatoid arthritis due to its antioxidant and anti-inflammatory activities.