The application of multiple anticancer drugs with a diverse mode of actions that target different cell components might lead to a desirable outcome to treat cancer types more effectively. A synergy of the drugs could lead to 1) increment in the effectiveness of therapy; 2) reduction in the dosage to decrease side effects while improving or maintaining efficacy; 3) mitigate or delay the development of resistance to the drugs. In this matter it is questionable what strategies and approaches are best for choosing candidates to be combined and tested even at the pre-clinical state of research. For anticancer drugs like Cisplatin (Cis-Pt) and Daunorubicin (Dauno), the main drug target is chromatin and both influence DNA topology; thus, the possibility arises that they may mutually influence each other’s binding to their chromatin targets. Therefore, we set out to determine how the Dauno evoked changes in chromatin structure and DNA topology affect the formation of Cis-Pt-DNA adducts, and vice versa how Cis-Pt influences Dauno binding. In this regard, we showed that Dauno and Cis-Pt antagonize one another through a decrease of ICLs in the presence of Dauno (at its low and high cc. alike), and also via the diminished Dauno uptake in the presence of Cis-Pt. They could synergize with each other through enhanced histone eviction by Dauno in the presence of co-treatment with Cis-Pt, accompanied by an increment of DNA bound anthracycline. These drug interactions have not been described before to the best of our knowledge and may impact cytotoxicity reached by combination treatment regimens in vivo. In the presence of Cis-Pt, Dauno concentration decreases in the cytoplasm (due to decreased uptake into the cell and/or the endosomal compartment). In the nucleus, its total amount is not affected by Dauno co-treatment, but the number of ICLs is decreased.