Abstract：Objective:To solve these uncertain threat that carrier materials and a new drug which is synthesized by drug-drug conjugate through covalent bond bring, insteading by more easy drug delivery system through non-covalent bond. Methods: A novel composite drug nanoparticles (DOX-Cb nanoparticles) have been prepared by reprecipitation after hydrophobic anticancer drug Doxorubicin (DOX) and hydrophobic anticancer drug chlorambucil (Cb) being mixed. Then using 1H nuclear magnetism(1H NMR), DLS,SEM,AFM ,TEM, Hoechst staining,western and confocal experiment confirm that DOX-Cb nanoparticles,which have antitumor activity in vitro, are prepared. Results: 1H NMR confirms that DOX and Cb compound is formed via electrostatic force between amino group and carboxyl group. DLS,SEM,AFM and TEM confirm that Composite drug nanoparticles have regular morphology and a narrow size distribution. Cytotoxicity test by using three tumor models（MCF-7，A549，HepG2） shows Composite drug nanoparticles have better effect of killing tumor cells than free DOX and Cb. Hoechst staining and western’result suggest that Composite drug nanoparticles kill tumor by inducing tumor cells apoptosis. confocal experiment verifys DOX-Cb nanoparticles are easier endocytosed by tumor cells than DOX. Conclusion: A novel composite drug nanoparticles (DOX-Cb nanoparticles) have been prepared by reprecipitation through electrostatic force between DOX and Cb, and composite drug nanoparticles were prepared by this strategy having better antitumor effect.
