A novel series of carbohydrate-based 1,2-naphthoquinones 13a–c and their substituted phenylhydrazono derivatives 14a–l were synthesized and evaluated for cytotoxicity against HL-60, MDA-MB435, HCT-116 and SF-295 cancer cell lines. The compounds 9a–c showed the best cytotoxicity profile (IC₅₀ below 2 μM) against HL-60 and MDA-MB 435 human cells, while the hydrazone derivative 14i (IC₅₀ HL-60 1.65 μM) was selective for leukemia when compared to the reference drug doxorubicin. None of the compounds exhibited lytic effects against mouse erythrocytes. Characterization of all compounds was confirmed by one- and two-dimensional NMR techniques (¹H, ¹³C-APT, COSY-¹H × ¹H and HETCOR ¹J_CH) and by elemental analysis.