A novel series of 2-arylcarbonylmethylthio-6-arylmethylpyrimidin-4(3H)-ones have been synthesized and evaluated for in vitro anti-HIV activities in MT-4 cells. Most of these new compounds showed moderate to potent activities against wild-type HIV-1 with an EC 50 range from 8.97μM to 0.010μM. Among them, the 6-(3,5-dimethylbenzyl) analogue 5p was identified as the most promising compound (EC 50 =0.010μM, SI>31,800) associated with moderate activity against the HIV-1 double mutant RT strain K103N+Y181C. The structure–activity relationships of these new congeners were further discussed.