Synthesis and characterization of well-defined amphiphilic block copolymers containing poly(2-ethyl-2-oxazoline) as hydrophilic block and poly(epsilon-caprolactone) or poly(L-lactide) as hydrophobic block is achieved by copper-catalyzed azide-alkyne cycloaddition (CuAAC) click chemistry. The clickable precursors, alpha-alkyne-functionalized poly(epsilon-caprolactone) and poly(L-lactide) and omega-azido-functionalized poly(2-ethyl-2-oxazoline) are simply prepared and joined using copper sulfate/ ascorbic acid catalyst system at room temperature. The structures of precursors and amphiphilic block copolymers are characterized by spectroscopic, chromatographic and thermal analyses. The cytotoxic activities of resulting amphiphilic block copolymers and their precursors are investigated in the prostate epithelial and cancer cells under in-vitro conditions. The treatment of the healthy prostate epithelial cell line PNT1A reveals that no significant cytotoxicity, whereas some significant toxic effects on the prostate cancer cell lines are observed.