Betulinic acid (BA), a triterpene in nature shows significant antitumor activity. But, its low solubility in aqueous phase leads to its poor bioavailability. The utilization of liposomal nanoformulation is a promising way to deliver BA with enhanced solubility profile and bioavailability. In present work, BA nanoliposomes were prepared and their physicochemical properties were optimized and characterized. BA nanoliposomes displayed optimum properties, with a particle size of 60-125 nm, zeta potential of -42.5 mV, and percent entrapment efficiency of 65.2% for drug delivery applications. The cytotoxic activity of BA loaded liposomal nanoformulation against A-549, MCF-7 and Hela cell lines confirmed its potential to hinder cancer cell proliferation. Hence, the present research work demonstrated the applicability of BA nanoliposomes as promising nanoformulation for cancer therapy.