Development of daunorubicin-loaded bovine serum albumin nanoparticles – preparation, characterisation, optimization of production process, investigation of in vitro and in vivo toxicity and activity

Albumin-based nanoparticles (NPs) represent promising drug delivery systems (DDSs). By incorporating cytostatic agents into bovine serum albumin nanoparticles (BSA-NPs), their intrinsic toxicity can be reduced, while the pharmacokinetic profile of drugs can be optimised. Daunorubicin (DAU) is an anthracycline derivative with potent antitumor activity through inhibition of topoisomerase II, is widely used to treat various types of leukemia. However, its clinical application is limited by severe cardiotoxic side effects, which may be significantly reduced through nanoparticle-based delivery. In the present study, DAU-containing BSA NPs were prepared using the desolvation method. The parameters of the desolvation process were optimised, and their effects on NP characteristics were systematically investigated. The dissolution profile, cellular uptake, and stability of the nanoparticles under different temperature, storage medium, and time conditions were evaluated. Under optimal preparation conditions, the resulting NPs exhibited and average particle size of 180-200nm and an encapsulation efficiency of 65-70 %. The cytotoxic effects of the prepared NPs were tested on four different cancer cell lines. The murine lymphoma P388 cell line demonstrated efficient uptake of both free DAU and DAU-containing BSA NPs after overnight incubation. The nanoparticles displayed sustained drug release at 37 °C and remained stable at 4 °C for 8 weeks in various storage solutions. In vivo toxicity revealed that the DAU-BSA nanoparticles exhibited reduced toxicity compared with free DAU. Based on these results, a standardized protocol was established for the preparation of daunorubicin-loaded BSA nanoparticles, enabling their use in future anticancer studies.