Herein, aminated mesoporous silica nanoparticles (AMPSNPs) were developed by using the co-condensation process and were further explored as a nanocarrier for disulfiram (DSF) delivery, which is classified as a “repurposed drug” for a variety of cancer therapies. The optimized DSF loading and encapsulation efficiency were found to be 18.05 ± 0.62% and 93.01 ± 0.83%, respectively. The scanning electron microscopy (SEM) showed a spherical and uniform formation of optimized AMPSNPs. Transmission electron microscopy (TEM) clearly displayed that the developed silica nanoparticles (NPs) contain mesostructures in them. For discovering a successful formation of an anticancer drug delivery system, various physicochemical characterization techniques were studied, such as PXRD, DSC, and FTIR. Hemolysis study revealed a better blood compatibility of the synthesized AMPSNPs. Furthermore, in vitro cellular cytotoxicity assay displayed an excellent cytotoxic profile of DSF-AMPSMPs compared to that of free DSF. Additionally, in vivo anti-tumor study conducted on A549 tumor-bearing nude mice verified a substantial tumor volume suppression and greater tumor growth inhibition with DSF-AMPSNP treatment group compared to using blank-AMPSNP and the free DSF group, with limited adverse-effects on mice health. Thus, we fervently have confidence that the AMPSNP could be utilized as a capable drug carrier for the delivery of DSF in cancer therapies.