2-Chloro-3-alkyl-1,4-naphthoquinone derivatives were synthesized and tested as the anti-acute myeloid leukaemia agents. The compound 9b (2-chloro-3-ethyl-5,6,7-trimethoxy-1,4-naphthoquinone) was the most potent toward HL-60 leukaemia cells. In mechanistic study for 9b, the protein levels of mtDNA-specific DNA polymerase γ (poly-γ) and mtDNA transcription factor A (mt-TFA) were decreased after the 24 h treatment, showing the occurrence of mtDNA damage. And 9b triggered cell cycle arrest at S phase accompanied by a secondary block in G2/M phase which had a direct link to the process of mtDNA damage. The dissipations of mitochondrial membrane potential and ATP also proceeded. On the other hand, 9b promoted the generation of ROS and resulted in the oxidation of intracellular GSH to GSSG. This process was coupled to the formation of adduct between 9b and GSH, detected by the UV–Vis spectrum and HRMS analysis. Depletion of GSH by buthionine sulfoximine enhanced ROS level and produced higher cytotoxicity, suggesting GSH was involved in the anti-leukemic mechanism of 9b. Together, our results provide new insights on the molecular mechanism of the derivatives of 2-chloro-1,4-naphthoquinone and 9b might be useful for the further development into an anti-leukemia agent.

合成了2-氯-3-烷基-1,4-萘醌衍生物，并将其作为抗急性髓样白血病药物进行了测试。化合物9b（2-氯-3-乙基-5,6,7-三甲氧基-1,4-萘醌）对HL-60白血病细胞最有效。在9b的机理研究中，处理24小时后mtDNA特异性DNA聚合酶γ（poly-γ）和mtDNA转录因子A（mt-TFA）的蛋白质水平降低，表明发生了mtDNA损伤。和9b中触发细胞周期停滞在伴随G2 / M期的二次块其中有一个直接链接到mtDNA损伤的过程中S期。线粒体膜电位和ATP的耗散也进行了。另一方面，9b促进了ROS的生成，并导致细胞内GSH氧化为GSSG。此过程与9b和GSH之间加合物的形成相关联，通过UV-Vis光谱和HRMS分析检测到。丁硫氨酸亚砜肟对GSH的消耗会增加ROS的水平并产生更高的细胞毒性，这表明GSH参与了9b的抗白血病机制。总之，我们的结果为2-氯-1,4-萘醌和9b衍生物的分子机理提供了新的见解，可能对进一步发展为抗白血病药物有用。