The purpose of this research was to create a binary nanodrug-delivery platform functionalized with aptamers (APs) and transferrin (Tf), and encapsulating daunorubicin (Drn) and luteolin (Lut) for leukemia therapy. Oligonucleotide ligands containing APs and Tf were designed and synthesized independently. AP-functionalized nanoparticles loaded with Drn (AP-Drn NPs) and Tf-functionalized nanoparticles loaded with Lut (Tf-Lut NPs) were fabricated through self-assembly. A binary nanodrug-delivery system co-functionalized with APs and Tf and co-loaded with Drn and Lut (AP/Tf-Drn/Lut NPs) was constructed via self-assembly of AP-Drn NPs and Tf-Lut NPs. The in vitro and in vivo performance of this system was assessed using a leukemia cell line and a tumor-bearing mouse model, compared to formulations decorated with a single ligand, loaded with a single drug, or free-drug combinations. The AP/Tf-Drn/Lut NPs were spherical with a nanoscale size (187.3 ± 5.3 nm) and exhibited drug-loading efficiency of approximately 85%. In vitro, the cytotoxicity of AP/Tf-Drn/Lut NPs was substantially greater than that of single-ligand-functionalized nanoparticles. The dual-drug-loaded AP/Tf-Drn/Lut NPs demonstrated superior inhibition of tumor cells compared to single-drug-loaded versions, indicating a synergistic effect between the two drugs. In vivo, AP/Tf-Drn/Lut NPs displayed the highest antileukemic efficacy with no observable toxicity. This study demonstrated that AP/Tf-Drn/Lut NPs represent a promising targeted drug-delivery platform for leukemia treatment, attributable to the synergistic action of the co-encapsulated drugs. Limitations of the system include stability challenges during scale-up production and translation from laboratory to clinical application.
