Combination therapy of kinases inhibitors and chemotherapeutics targeting tubulin dynamics is an important strategy to improve therapeutic efficacy and overcome the resistance to single-target drug therapies. Inspired by this, we report herein the rational design of 3,5-disubsituted-1H-pyrazolo[3,4-b]pyridines as multiacting molecules that are capable of inhibiting tubulin and kinases simultaneously. Among them, 8g showed excellent antiproliferative activities toward a panel of cancer cell lines. 8g strongly inhibited tubulin assembly and demonstrated a potent inhibition toward FLT3 and Abl1 in both enzymatic and cellular assays. 8g caused a cell cycle arrest at G2/M phase, and significantly disrupted HUVEC tube formation. In vivo efficacy studies showed that 8g significantly inhibited tumor growth on the K562 leukemia xenograft model at 10 mg/kg. Collectively our studies suggest that the excellent antiproliferative potency of 8g may be attributed to its potent inhibitory activity against both microtubule and kinases.

激酶抑制剂和针对微管蛋白动力学的化疗药物的联合治疗是提高治疗效果和克服单靶点药物治疗耐药性的重要策略。受此启发，我们在此报告了 3,5-二取代-1H-吡唑并[3,4-b]吡啶作为能够同时抑制微管蛋白和激酶的多作用分子的合理设计。其中，8g对一组癌细胞系表现出优异的抗增殖活性。8g强烈抑制微管蛋白组装，并在酶和细胞测定中显示出对 FLT3 和 Abl1 的有效抑制。8g导致细胞周期停滞在G2/M期，并显着破坏HUVEC管形成。体内功效研究表明，8g 10 mg/kg 可显着抑制 K562 白血病异种移植模型上的肿瘤生长。总的来说，我们的研究表明，8g优异的抗增殖效力可能归因于其对微管和激酶的有效抑制活性。