Bioactive peptide with anti-proliferative activity is a novel prospect to develop cancer drugs with fewer or no side effects, as well as cost benefits. The current review provides a development strategy for anti-proliferative peptides derived from fish and its discards through fish protein hydrolysate. The study indicates that enzymatic hydrolysates act as a superior anti-proliferative agent through superior enzyme selection. In general, ion-exchange chromatography, RP-HPLC, ultra, and gel filtration methods are used to purify bioactive peptides. Characterization of the purified peptide can be done with the help of cytotoxicity assay, molecular mass, amino acid composition, and sequence. Most studies on anti-proliferative bioactive peptides have been carried out via in vitro analysis. However, very few studies have provided confirmatory results through in vivo analysis.