Steiner Ivana, Stojanovic Nikolina, Bolje Aljosa, Brozovic Anamaria, Polancec Denis, Ambriovic-Ristov Andreja, Stojkovic Marijana Radic, Piantanida Ivo, Eljuga Domagoj, Kosmrlj Janez, Osmak Maja
Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia.
Faculty of Chemistry and Chemical Technology, University of Ljubljana, Slovenia.
Radiol Oncol. 2016 Jul 19;50(3):280-8. doi: 10.1515/raon-2016-0027. eCollection 2016 Sep 1.
In order to increase the effectiveness of cancer treatment, new compounds with potential anticancer activities are synthesized and screened. Here we present the screening of a new class of compounds, 1-(2-picolyl)-, 4-(2-picolyl)-, 1-(2-pyridyl)-, and 4-(2-pyridyl)-3-methyl-1,2,3-triazolium salts and 'parent' 1,2,3-triazole precursors.
Cytotoxic activity of new compounds was determined by spectrophotometric MTT assay on several tumour and one normal cell line. Effect of the selected compound to bind double stranded DNA (ds DNA) was examined by testing its influence on thermal stability of calf thymus DNA while its influence on cell cycle was determined by flow cytometric analysis. Generation of reactive oxygen species (ROS) was determined by addition of specific substrate 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA).
Parent triazoles were largely inactive, while some of the triazolium salts were highly cytotoxic for HeLa cells. Triazolium salts exhibited high cell-type dependent cytotoxicity against different tumour cells. Selected compound (4-(4-methoxyphenyl)-3-methyl-1-(2-picolyl)-1H-1,2,3-triazolium hexafluorophosphate(V) (2b) was significantly more cytotoxic against tumour cells than to normal cells, with very high therapeutic index 7.69 for large cell lung carcinoma H460 cells. Additionally, this compound was similarly cytotoxic against parent laryngeal carcinoma HEp-2 cells and their drug resistant 7T subline, suggesting the potential of this compound in treatment of drug resistant cancers. Compound 2b arrested cells in the G1 phase of the cell cycle. It did not bind ds DNA, but induced ROS in treated cells, which further triggered cell death.
Our results suggest that the 'click' triazolium salts are worthy of further investigation as anti-cancer agents.
为提高癌症治疗效果，合成并筛选了具有潜在抗癌活性的新化合物。在此，我们展示了一类新化合物的筛选，即1-(2-吡啶甲基)-、4-(2-吡啶甲基)-、1-(2-吡啶基)-和4-(2-吡啶基)-3-甲基-1,2,3-三唑鎓盐以及“母体”1,2,3-三唑前体。
通过分光光度法MTT试验测定新化合物对几种肿瘤细胞系和一种正常细胞系的细胞毒性活性。通过测试其对小牛胸腺DNA热稳定性的影响来检测所选化合物与双链DNA（ds DNA）结合的效果，同时通过流式细胞术分析确定其对细胞周期的影响。通过添加特异性底物5-(及-6)-氯甲基-2',7'-二氯二氢荧光素二乙酸酯、乙酰酯（CM-H2DCFDA）来测定活性氧（ROS）的产生。
母体三唑大多无活性，而一些三唑鎓盐对HeLa细胞具有高度细胞毒性。三唑鎓盐对不同肿瘤细胞表现出高度的细胞类型依赖性细胞毒性。所选化合物（4-(4-甲氧基苯基)-3-甲基-1-(2-吡啶甲基)-1H-1,2,3-三唑鎓六氟磷酸盐(V)（2b）对肿瘤细胞的细胞毒性明显高于对正常细胞的毒性，对大细胞肺癌H460细胞的治疗指数高达7.69。此外，该化合物对喉癌亲本HEp-2细胞及其耐药7T亚系具有相似的细胞毒性，表明该化合物在治疗耐药癌症方面具有潜力。化合物2b使细胞停滞于细胞周期的G1期。它不结合ds DNA，但在处理后的细胞中诱导ROS产生，进而引发细胞死亡。
我们的结果表明，“点击”三唑鎓盐作为抗癌剂值得进一步研究。
