A direct C-H trifluoromethylation of 2-amino-1,4-naphthoquinone analogues is described. This reaction proceeds under mild conditions at open atmosphere, providing a range of CF3-containing naphthoquinones with good yield and functional group compatibility. All synthetic compounds were screened for antiproliferative activity against three human cancer cell lines. Notably, some of those trifluoromethyl analogs, such as 3a, 3g, 3j, and 3t, showed good antiproliferative profiles.