The aim of this paper was to provide the physico-chemical characterization of a key process leading to amplification of the antitumor effect of antibiotic Doxorubicin (Dox) in vivo and in vitro and occurring at the molecular level through complexation with C60 fullerene. A wide range of physico-chemical tools was used such as UV/Vis and NMR spectroscopies, atomic force microscopy, isothermal titration calorimetry and zeta-potential methods. The unusual thermodynamic behavior of the complexation process was reported, featuring unexpected and, to a certain extent, contradictory experimental observations. The explanation of the obtained results was proposed resulting in creation of a general view on aromatic drug binding with C60 fullerene. Based on these results some important practical outcomes for anticancer therapy were formulated.
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