Diacetylene moiety, DIOL was functionalized to chemotherapeutic drug, chlorambucil and water-soluble moiety poly(ethylene glycol) to get our macromonomer MPDC. MPDC was further cyclopolymerized using Hoveyda Grubbs’ second generation catalyst, results to form polymer PMPDC. Amphiphilic nature of PMPDC leads to self-assembly in water which was characterized by dynamic light scattering and transmission electron microscope. Cytotoxicity study was performed with HeLa and MCF 7 cancer cell lines which showed good cell growth inhibition. As PMPDC having non-fluorescent property, another polymer PMPDP was synthesized as control using 1-pyrenebutyric acid which was further used for drug release at different pH and cell internalization study. Increase in internalization of polymer with increase in concentration was observed in both HeLa and MCF 7 cells. Formation of all the compounds were characterized carefully by different analytical techniques.