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A novel series of <em>N</em>-methylpicolinamide-4-thiol derivatives were synthesized and evaluated on human cancer cell lines. Among them, compound <strong>6p</strong> displayed potent and<strong> </strong>broad-spectrum anti-proliferative activities<em> in</em><em> vitro</em> on some human cancer cell lines, even better than sorafenib. The advanced kinase inhibitory assays showed that compound <strong>6p</strong> could selectively inhibit Aurora-B kinase. The biological results were rationalized by the molecular docking study, which indicated the stable interactions of <strong>6p</strong> with the Aurora-B kinase.