Brain cancer is the second leading cause of death after leukemia for children. In this study, brain cancer cells (U-87MG) were cultured with 10 % FBS. The Macrovipera lebetina venom was purified by two methods of chromatography. Anti-adhesion and cytotoxic effects were investigated by trypan blue and MTT assay. Six fractions were obtained from gel filtration chromatography. Fraction N.5 had anti-adhesion and cytotoxic effects, it was selected for ion exchange chromatography. The effect of four fractions obtained from ion exchange chromatography on the brain cells and control cell was studied and fraction N.3 had the highest level of cytotoxicity. Anti-adhesion and fatal effects on the brain and control cells were estimated to be 26 and 14 %, respectively. In this study, the active fraction had a cytotoxic effect on the brain cancer cell line.