Chemotherapeutics approaches used to cure cancer are frequently faced a problem of drug resistances. Secondary metabolites of the plants having anticancer potency can be effectively raised by formulating them at nanoscale. Acronine is a natural alkaloid with an acridine structure isolated from the bark of the plant Acronychia baueri (Australian scrub ash) with antineoplastic properties. Acronycine is an antitumor alkaloid with poor water solubility and low potency. Tamoxifen is a selective estrogen receptor modulator used to treat estrogen receptor positive breast cancer, reduce the risk of invasive breast cancer following surgery, or reduce the risk of breast cancer in high risk women. To improve the bioavailability and to generate synergetic effect, Acronine and TAM loaded Chitosan nanoparticles (ATCNPs) were synthesized by oil in oil (O/O) emulsion solvent evaporation techniques. The zeta potential value of ATCNPs was come out to be -45.9 mV representing relative stability of nanoparticles. The percentage encapsulation efficiency value was found to be 78.1% for Acronine and 78.4% for TAM. The ATCNPs possess particle size in the range of 55 – 78nm as revealed by TAM. SEM image analysis confirmed that the nanoparticles have spherical in shapes. The ATCNPs exhibited sustained release and their anti-oxidant and anti-cancer potency was much more pronounced than free Acronine and TAM particles alone. The in vitro studies demonstrated that Acronine and TAM encapsulated in dammar gum inhibited growth of A-549 cells, MCF-7 and Hela cell lines, respectively more effectively than Acronine and TAM alone which proved their robust anticancer potential.