Novel indazole and benzimidazole analogues were designed and synthesized as tubulin inhibitors with potent antiproliferative activities. Among them, compound <b>12b</b> exhibited the strongest inhibitory effects on the growth of cancer cells with an average IC<sub>50</sub> value of 50 nM, slightly better than colchicine. <b>12b</b> exhibited nearly equal potency against both, a paclitaxel-resistant cancer cell line (A2780/T, IC<sub>50</sub> = 9.7 nM) and the corresponding parental cell line (A2780S, IC<sub>50</sub> = 6.2 nM), thus effectively overcoming paclitaxel resistance <i>in vitro</i>. The crystal structure of <b>12b</b> in complex with tubulin was solved to 2.45 Å resolution by X-ray crystallography, and its direct binding was confirmed to the colchicine site. Furthermore, <b>12b</b> displayed significant <i>in vivo</i> antitumor efficacy in a melanoma tumor model with tumor growth inhibition rates of 78.70% (15 mg/kg) and 84.32% (30 mg/kg). Collectively, this work shows that <b>12b</b> is a promising lead compound deserving further investigation as a potential anticancer agent.