Apoferritin-based nanomedicine platform for drug delivery: equilibrium binding study of daunomycin with DNA

Abstract

This paper describes the preparation and optimization of the analytical properties of the protein based drug delivery platform apoferritin. In biological systems, the protein cage ferritin is used to store iron and to keep it from building to toxic levels in cells. When the iron atoms are removed from ferritin, apoferritin is formed. In this study, daunomycin, an anthracycline antibiotic drug that has been used for specific types of cancer treatment such as acute myeloid leukemia and acute lymphocytic leukemia, was encapsulated within the protein cage for drug delivery. Daunomycin slows or stops the growth of cancer cells by binding with the cell's DNA. The model for daunomycin-DNA complex binding mechanism is intercalation, where daunomycin binds with approximately every 3 base pairs causing a local unwinding, but a negligible distortion of the helical conformation. The binding affinity for free DNA is higher than that of structured DNA in cells. Upon binding with DNA the fluorescence intensity of daunomycin decreases. We used apoferritin's ability to disassemble and reassemble under pH control to load the therapeutic compound daunomycin. The combination of a modifiable interior and exterior surface and the passable hydrophobic and hydrophilic channels through the cage allows the containment or attachment of both insoluble and soluble drugs for delivery. At experimental pH 5 conditions the interaction between the apoferritin interior cage and daunomycin is weak making it difficult to encapsulate the drug effectively within the protein cage. The incorporation of poly-L-aspartic acid (PLAA), a polypeptide and biodegradable material that does not increase the toxicity of the drug delivery system and is negatively charged at pH 5.0, into the drug delivery system resulted in a substantial improvement in the drug encapsulation. The binding properties of free daunomycin with DNA were compared to the newly synthesized apoferritin protein based drug delivery system. Encapsulation of the daunomycin within the apoferritin protein cage had little effect upon the intrinsic binding constant, K_i, or the exclusion parameter n as compared to the free daunomycin model. The study resulted in the design and optimization of a unique protein based drug delivery platform using the protein cage apoferritin for potential therapeutic administration of the anti-cancer agent daunomycin.