With the development of tyrosine kinase inhibitor (TKI) resistance, finding the novel effective chemotherapeutic agent is of seminal importance for chronic myelogenous leukemia (CML) treatment. This study aims to find the effective anti-leukemic candidates and investigate the possible underlying mechanism. We synthesized the novel coumarin derivatives and evaluated their anti-leukemic activity. Cell viability assay revealed that compound DBH2 exhibited the potent inhibitory activity on the proliferation of CML K562 cells and TKI resistant K562 cells. Morphological observation and flow cytometry confirmed that DBH2 could selectively induce cell apoptosis and cell cycle arrest at G2/M phase of the K562 cells, which was further confirmed on the bone marrow cells from CML transgenic model mice and CD34⁺ bone marrow leukemic cells from CML patients. Treatments of DBH2 in combination with imatinib could prolong the survival rate of SCL-tTA-BCR/ABL transgenic model mice significantly. Quantitative RT-PCR revealed that DBH2 inhibited the expression of STAT3 and STAT5 in K562 cells, and caspase-3 knockout alleviated the DBH2 induced apoptosis. Furthermore, DBH2 could induce the expression of PARP1 and ROCK1 in K562 cells, which may play the important role in caspase-dependent apoptosis. Our results concluded that coumarin derivative DBH2 serves as a promising candidate for the CML treatment, especially in the combination with imatinib for the TKI resistant CML, and STAT/caspase-3 pathway was involved in the molecular mechanism of anti-leukemic activity of DBH2.

随着酪氨酸激酶抑制剂 (TKI) 耐药性的发展，寻找新型有效的化疗药物对慢性粒细胞白血病 (CML) 的治疗具有重要意义。本研究旨在寻找有效的抗白血病候选物并研究可能的潜在机制。我们合成了新型香豆素衍生物并评估了它们的抗白血病活性。细胞活力测定显示化合物 DBH2 对 CML K562 细胞和 TKI 耐药 K562 细胞的增殖表现出有效的抑制活性。形态学观察和流式细胞术证实DBH2可选择性诱导K562细胞凋亡和细胞周期停滞在G2/M期，这在CML转基因模型小鼠和CD34 + 骨髓细胞中得到进一步证实^。来自 CML 患者的骨髓白血病细胞。DBH2联合伊马替尼治疗可显着延长SCL-tTA-BCR/ABL转基因模型小鼠的存活率。定量 RT-PCR 显示 DBH2 抑制 K562 细胞中 STAT3 和 STAT5 的表达，caspase-3敲除减轻 DBH2 诱导的细胞凋亡。此外，DBH2 可诱导 K562 细胞中 PARP1 和 ROCK1 的表达，这可能在 caspase 依赖性细胞凋亡中发挥重要作用。我们的结果得出结论，香豆素衍生物 DBH2 作为 CML 治疗的有希望的候选者，特别是在与伊马替尼联合治疗 TKI 耐药 CML 时，STAT/caspase-3 通路参与 DBH2 抗白血病活性的分子机制。