AbstractThe antiviral activities of antileukemic drugs 1‐β‐D‐arabinofuranosylcytosine (Cytarabine; Ara‐C), 2,2′‐anhydro‐1‐β‐D‐arabinofuranosylcytosine (Ancitabine; Cyclo‐C), and N4‐behenoyl‐1‐β‐D‐arabinofuranosylcytosine (Enocitabine; BH‐AC) were evaluated in vitro against human cytomegalovirus (HCMV) in comparison with those of five other antiviral drugs. Both Ara‐C and Cyclo‐C showed the strongest inhibitory effect to HCMV. BH‐AC inhibited the replication of HCMV and depicted almost as the same dose‐response curve as Ganciclovir (DHPG). In the presence of Ara‐C, Cyclo‐C, or BH‐AC, triphosphate forms of the nucleoside analogs were detected in the HCMV‐infected cells, and synthesis of HCMV DNA was strongly suppressed. Thus, Ara‐C, Cyclo‐C, and BH‐AC were not only antileukemic, but also antiviral in vitro. However, Ara‐C and Cyclo‐C may not be suitable as anti‐HCMV agents, because they are cytotoxic or excreted rapidly in the urine in vivo [Van Voris, 1984; Hirayama et al., 1974]. Because of lower toxicity and longer retention in vivo, BH‐AC may be expected as an anti‐HCMV agent in patients with leukemia, in addition to serving as an antileukemic drug.