DNA Topoisomerases and their poisoning

Topoisomerases are ubiquitous enzymes involved in maintaining genomic stability of the cell by modulating the level of supercoiling of DNA strands. Besides their customary functions, topoisomerases are important cellular targets of widely used anticancer drugs. In particular, topoisomerase IIα (Top2α) has been postulated as the primary molecular target of anthracycline’s anticancer activity (Mordente et al. 2017). Despite their wide use in chemotherapy, amthracyclines have a limiting side effect. In particular the cardiotoxicity is the most frequent adverse effect of both conventional and modern anticancer-targeted therapy, representing the leading noncancer-related cause of morbidity and mortality in long-term survivors (Hamo et al. 2015).

Coupling the drug to peptide carrier could improve solubility, decrease side effects and can offer the possibility of cell or tissue specific introduction (Hudecz et al. 1992; Duncan et al. 1987). Another approach for intracellular delivery is using cell-penetrating peptides like oligoarginines with cationic character; this method is less specific but could be highly efficient (Nakase et al. 2008;

Futaki et al. 2005).

In my work, I analyzed the biochemical properties of a new oligoarginine-daunomycin conjugate previously described by Miklàn et al. (2009). It’s known that anticancer activity of Daunomycin is based on both DNA binding properties and inhibition of human topoisomerase IIα. So I investigated the inhibition effect of Arg-derivative on both human topoisomerase IIα and topoisomerase I and I

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found that it is able to inhibit both topoisomerases relaxation activity efficiently as Daunomycin.

Then I analyzed the DNA binding properties of Daunomycin conjugate and I demonstrated that it is able to interact with DNA mainly by intercalative mode as already known for the free drug.

Furthermore Arg-derivative is also able to stabilize the double helical structure of DNA as demonstrated by melting experiment. These results suggest that oligoarginine-daunomycin conjugate could be a promising alternative to Daunomycin in antitumor therapy.

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