Sztanke Krzysztof, Pasternak Kazimierz, Rzymowska Jolanta, Sztanke Małgorzata, Kandefer-Szerszeń Martyna
Chair and Department of Medical Chemistry, Professor Feliks Skubiszewski Medical University, 4 Staszica Street, 20-081 Lublin, Poland.
Eur J Med Chem. 2008 May;43(5):1085-94. doi: 10.1016/j.ejmech.2007.07.009. Epub 2007 Jul 29.
Synthesis, structure elucidation and anticancer activities of novel fused 1,2,4-triazine aryl derivatives containing the ethoxycarbonyl (6-10) and carbohydrazide formations (11-15) are presented. Molecular structures of the synthesized compounds were confirmed by IR, (1)H NMR, (13)C NMR, EI-MS spectra and elemental analyses. Antitumour activities in vitro for heterobicyclic hydrazides of the type 11-14 were evaluated by BrdU method for human LS180, SiHa and T47D carcinoma cells. Amongst them, hydrazide 14 has exhibited remarkable inhibitory effect against SiHa and LS180 tumour cells, and simultaneously was found to be non-toxic towards the human normal cell line-HSF cells. Furthermore, the pulse field gel electrophoresis experiment was performed for characterizing DNA-cleaving activity of heterobicycle 14. The DNA fragments of 2500, 2000 and 500 kilobase pairs (kbp) were commonly detected in the cancer cell lines (SiHa, LS180 and T47D) treated with compound 14. DNA fragmentation pattern, since three types of fragments induced by the tested hydrazide of the type 14 were detected, suggesting a way of interaction with DNA. It is worth pointing out, that DNA strand breaks were also produced in human breast cancer (T47D) cells, a cell line where the induction of DNA fragmentation is very difficult. Moreover, the statistically significant apoptotic activity in T47D human breast cancer cells for the tested heterobicycle 14 was proved using the annexin V-binding assay. The antiproliferative properties in vitro for compounds 6-14 were evaluated by MTT method for human leukaemic Jurkat cells. Significant viability decreases in Jurkat cells treated with different concentrations of compounds 10 and 11 were observed, suggesting that these derivatives have antiproliferative activities. Their acute toxicities were established. For these compounds the influence on the central nervous system of mice in behavioural tests was examined. Molecular structure for free base of the intermediate 4 was confirmed by (1)H-(1)H COSY, HMBC and HMQC correlations.
本文介绍了含乙氧羰基（6 - 10）和碳酰肼结构（11 - 15）的新型稠合1,2,4 - 三嗪芳基衍生物的合成、结构解析及抗癌活性。通过红外光谱、（1）H核磁共振、（13）C核磁共振、电子轰击质谱（EI - MS）光谱和元素分析确定了合成化合物的分子结构。采用BrdU法对人LS180、SiHa和T47D癌细胞评估了11 - 14型杂环酰肼的体外抗肿瘤活性。其中，酰肼14对SiHa和LS180肿瘤细胞表现出显著的抑制作用，同时发现对人正常细胞系 - HSF细胞无毒。此外，进行了脉冲场凝胶电泳实验以表征杂环化合物14的DNA切割活性。在用化合物14处理的癌细胞系（SiHa、LS180和T47D）中通常检测到2500、2000和500千碱基对（kbp）的DNA片段。由于检测到由14型受试酰肼诱导的三种类型的片段，DNA片段化模式表明了其与DNA的相互作用方式。值得指出的是，在人乳腺癌（T47D）细胞中也产生了DNA链断裂，该细胞系中诱导DNA片段化非常困难。此外，使用膜联蛋白V结合试验证明了受试杂环化合物14在T47D人乳腺癌细胞中具有统计学上显著的凋亡活性。采用MTT法对人白血病Jurkat细胞评估了化合物6 - 14的体外抗增殖特性。观察到用不同浓度的化合物10和11处理的Jurkat细胞的活力显著降低，表明这些衍生物具有抗增殖活性。确定了它们的急性毒性。在行为测试中检查了这些化合物对小鼠中枢神经系统的影响。通过（1）H - （1）H化学位移相关谱（COSY）、异核多键相关谱（HMBC）和异核单量子相关谱（HMQC）相关性确定了中间体4游离碱的分子结构。
