We observed a synergistic antiproliferation effect with combined use of a DNA minor groove binder and a histone deacetylase (HDAC) inhibitor. Inspired by this result, a new series of benzimidazole<b>-</b>hydroxamate hybrids were designed and synthesized to target both DNA minor groove and HDAC. The most active compounds <b>9k</b> and <b>9l</b> not only exhibited improved HDAC inhibitory activities compared to SAHA but also possessed potent antiproliferation activities against tumor cells. Importantly, compounds <b>9k</b> and <b>9l</b> showed good in vivo antitumor efficacies in both HEL xenograft model and murine melanoma model. We also found that <b>9k</b> and <b>9l</b> promote the antigen presentation and activate T cells, thereby triggering antitumor immunity. Moreover, these inhibitors reshaped the tumor immune microenvironment by inhibiting the recruitment of Treg cells and promoting the polarization of tumor-infiltrating macrophages to M2 type with antitumor activity. Our study validated the effectiveness of incorporating a DNA-binding fragment in HDAC inhibitors as novel multitargeting antitumor agents.