Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University , Riyadh 11451, Saudi Arabia.
J Med Chem. 2014 Nov 13;57(21):9139-51. doi: 10.1021/jm501263m. Epub 2014 Nov 5.
Fluorescein hydrazones (3a-3l) were synthesized in three steps with 86-91% overall yields. Topo I- and IIα-mediated relaxation and cell viability assay were evaluated. 3d inhibited 47% Topo I (camptothecin, 34%) and 20% Topo II (etoposide 24%) at 20 μM. 3l inhibited 61% Topo II (etoposide 24%) at 20 μM. 3d and 3l were further evaluated to determine their mode of action with diverse methods of kDNA decatenation, DNA-Topo cleavage complex, comet, DNA intercalating/unwinding, and Topo IIα-mediated ATP hydrolysis assays. 3d functioned as a nonintercalative dual inhibitor against the catalytic activities of Topo I and Topo IIα. 3l acted as a Topo IIα specific nonintercalative catalytic inhibitor. 3d activated apoptotic proteins as it increased the level of cleaved capase-3 and cleaved PARP in a dose- and time-dependent manner. The dose- and time-dependent increase of G1 phase population was observed by treatment of 3d along with the increase of p27(kip1) and the decrease of cyclin D1 expression.
荧光素腙（3a-3l）通过三步合成，总收率为 86-91%。评估了拓扑异构酶 I 和 IIα 的松弛和细胞活力测定。3d 在 20μM 时抑制 47%的拓扑异构酶 I（喜树碱，34%）和 20%的拓扑异构酶 II（依托泊苷 24%）。3l 在 20μM 时抑制 61%的拓扑异构酶 II（依托泊苷 24%）。进一步评估 3d 和 3l，以确定其作用模式，包括不同的 kDNA 解连环、DNA-拓扑异构酶切割复合物、彗星、DNA 嵌入/解旋以及拓扑异构酶 IIα 介导的 ATP 水解测定。3d 作为一种非嵌入性双重抑制剂，针对拓扑异构酶 I 和拓扑异构酶 IIα 的催化活性。3l 作为拓扑异构酶 IIα 的特异性非嵌入性催化抑制剂。3d 通过增加切割的 caspase-3 和切割的 PARP 的水平，以剂量和时间依赖性方式激活凋亡蛋白。通过用 3d 处理观察到 G1 期细胞群的剂量和时间依赖性增加，同时 p27(kip1)增加和 cyclin D1 表达减少。
