2′-C-Cyano-2′-deoxy-1-β-D-arabinofuranosylcytosine (CNDAC): A Mechanism-Based DNA-Strand-Breaking Antitumor Nucleoside¹

Abstract

The antitumor mechanism of action of 2′-C-cyano-2′-deoxy-1-β-d-arabinofuranosylcytosine (CNDAC) has been examined. CNDAC was designed as a potentially DNA-self-strand-breaking nucleoside. It had potent antitumor effects against various solid tumors in vitro as well as in vivo. Using a chain-extension method with Vent (exo⁻) DNA polymerase and a short primer/template system, we found that 5′-triphosphate of CNDAC (CNDACTP) was incorporated into the primer at a site opposite a guanine residue in the template. After further chain-extension reaction of the primer containing CNDAC at the 3′-terminus, chain elongation was not observed. Therefore, CNDACTP appeared to act as a chain-terminator. Analyses of the structure of the 3′-terminus in the primer revealed 2′-C-cyano-2′,3′-didehydro-2′,3′-dideoxycytidine (ddCNC) together with CNDAC and 2′-C-cyano-2′-deoxy-1-β-d-ribofuranosylcytosine (CNDC). The existence of ddCNC in the 3′-end of the primer would be due to the self-strand-break by the nucleotide incorporated next to CNDAC. We also found that CNDAC was epimerized to CNDC in near-neutral to alkaline media. Therefore, CNDC found in the primer was epimerized after incorporation of CNDACTP into the primer. We also described the metabolism of CNDAC.