University of Kragujevac, Faculty of Science, Department of Chemistry, Radoja Domanovića 12, 34000 Kragujevac, Serbia.
University of Kragujevac, Faculty of Medical Sciences, Centre for Molecular Medicine and Stem Cell Research, Svetozara Markovića 69, 34000 Kragujevac, Serbia.
Bioorg Chem. 2019 Jul;88:102954. doi: 10.1016/j.bioorg.2019.102954. Epub 2019 Apr 27.
In order to make a progress in discovering a new agents for chemotherapy with improved properties and bearing in mind the fact that substituted 3-hydroxy-3-pyrrolin-2-ones belong to a class of biologically active compounds, series of novel 1,5-diaryl-4-(2-thienylcarbonyl)-3-hydroxy-3-pyrrolin-2-ones were synthesized and characterized by spectral (UV-Vis, IR, NMR, ESI-MS), X-ray and elemental analysis. All compounds were examined for their cytotoxic effect on human cancer cell lines HeLa and MDA-MB 231 and normal fibroblasts (MRC-5). Four compounds, 3-hydroxy-1-(p-tolyl)-4-(2-thienylcarbonyl)-5-(4-chlorophenyl)-2,5-dihydro-1H-pyrrol-2-one (D10), 3-hydroxy-1-(3-nitrophenyl)-4-(2-thienylcarbonyl)-5-(4-(benzyloxy)phenyl)-2,5-dihydro-1H-pyrrol-2-one (D13), 3-hydroxy-1-(4-nitrophenyl)-4-(2-thienylcarbonyl)-5-(4-(benzyloxy)phenyl)-2,5-dihydro-1H-pyrrol-2-one (D14), and 3-hydroxy-1-(4-chlorophenyl)-4-(2-thienylcarbonyl)-5-(4-(benzyloxy)phenyl)-2,5-dihydro-1H-pyrrol-2-one (D15), that showed the highest cytotoxicity against malignant cells and the best selectivity towards normal cells were selected for further experiments. Results obtained by investigating mechanisms of cytotoxic activity suggest that selected 3-hydroxy-3-pyrrolin-2-one derivatives in HeLa cells induce apoptosis that is associated with S phase arrest (D13, D15, and D10) or unrelated to cell cycle distribution (D14). Additionally, to better understand their suitability for potential use as anticancer medicaments we studied the interactions between biomacromolecules (DNA or BSA) and D13 and D15. The results indicated that D13 and D15 have great affinity to displace EB from the EB-DNA complex through intercalation [K = (3.7 ± 0.1) and (3.4 ± 0.1) × 10 M, respectively], an intercalative mode also confirmed through viscosity measurements. K values, obtained as result of fluorescence titration of BSA with D13 and D15 [K = (4.2 ± 0.2) and (2.6 ± 0.2) × 10 M, respectively], support the fact that a significant amount of the tested compounds could be transported and distributed through the cells. In addition, by DNA and BSA molecular docking study for D13, D14 and D15 is determined and predicted the binding mode and the interaction region.
为了在具有改进性质的新化疗药物的发现方面取得进展，并考虑到取代的 3-羟基-3-吡咯啉-2-酮属于一类具有生物活性的化合物这一事实，我们合成了一系列新型的 1,5-二芳基-4-(2-噻吩羰基)-3-羟基-3-吡咯啉-2-酮，并通过光谱（UV-Vis、IR、NMR、ESI-MS）、X 射线和元素分析对其进行了表征。所有化合物均对人癌细胞系 HeLa 和 MDA-MB 231 以及正常成纤维细胞（MRC-5）进行了细胞毒性测试。四种化合物，3-羟基-1-（对甲苯基）-4-（2-噻吩羰基）-5-（4-氯苯基）-2,5-二氢-1H-吡咯-2-酮（D10）、3-羟基-1-（3-硝基苯基）-4-（2-噻吩羰基）-5-（4-（苯甲氧基）苯基）-2,5-二氢-1H-吡咯-2-酮（D13）、3-羟基-1-（4-硝基苯基）-4-（2-噻吩羰基）-5-（4-（苯甲氧基）苯基）-2,5-二氢-1H-吡咯-2-酮（D14）和 3-羟基-1-（4-氯苯基）-4-（2-噻吩羰基）-5-（4-（苯甲氧基）苯基）-2,5-二氢-1H-吡咯-2-酮（D15）对恶性细胞表现出最高的细胞毒性和对正常细胞的最佳选择性，因此被选为进一步实验的对象。通过研究细胞毒性作用的机制得出的结果表明，所选的 3-羟基-3-吡咯啉-2-酮衍生物在 HeLa 细胞中诱导的凋亡与 S 期阻滞（D13、D15 和 D10）有关，或与细胞周期分布无关（D14）。此外，为了更好地了解它们作为抗癌药物的适用性，我们研究了生物大分子（DNA 或 BSA）与 D13 和 D15 之间的相互作用。结果表明，D13 和 D15 通过嵌入从 EB-DNA 复合物中置换 EB 的能力非常强[K 值分别为（3.7±0.1）和（3.4±0.1）×10M]，这一嵌入模式也通过粘度测量得到了证实。通过 D13 和 D15 对 BSA 的荧光滴定获得的 K 值[K 值分别为（4.2±0.2）和（2.6±0.2）×10M]表明，大量测试化合物可以通过细胞运输和分布。此外，通过 D13、D14 和 D15 的 DNA 和 BSA 分子对接研究，确定并预测了它们的结合模式和相互作用区域。
