Ahn Seunghyun, Lee Youngshim, Park Jihyun, Lee Junho, Shin Soon Y, Lee Young H, Koh Dongsoo, Lim Yoongho
Division of Bioscience and Biotechnology, BMIC, Konkuk University, Seoul 05029, Korea.
Department of Biological Sciences, Konkuk University, Seoul 05029, Korea.
Med Chem. 2018;14(8):851-862. doi: 10.2174/1573406414666180418143048.
The Hantzsch ester, diethyl 2,6-dimethyl-1,4-dihydropyridine-3,5- dicarboxylate, has been used as a hydride donor and its various biological effects have been reported. To identify chemotherapeutic agents with apoptotic effects, 21 diethyl 2,6-dimethyl-1,4- dihydropyridine-3,5-dicarboxylates were designed and synthesized; they have not been reported as apoptosis inducers thus far. Their structure-cytotoxicity relationships were investigated. Further biological experiments were performed on the title compound.
The cytotoxicities of the current synthetic compounds were measured using a clonogenic assay in HCT116 human colon cancer cells. An annexin V staining assay was used to confirm if the title compound induced apoptosis. To identify the synthetic compounds, Nuclear Magnetic Resonance (NMR) spectroscopy and high-resolution mass spectrometry (HR-MS) were conducted. As molecular symmetry was observed in the NMR spectroscopic data, the three dimensional structures were determined from ab initio calculations and X-ray crystallography.
The results obtained from NMR spectroscopy, ab initio calculations, and X-ray crystallography revealed that the diethyl 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylate derivatives synthesized in this research have symmetric structures. The cytotoxicities of the 21 derivatives were tested in the HCT116 human colon cancer cell lines, and their half-maximal cell growth inhibitory concentrations ranged between 16.29 and 68.88 µM. Structure-cytotoxicity relationships demonstrated that bulky substitutions were preferred, para-positioned substituents tended to have better cytotoxic values, and the polarity may have a function as well. The cytotoxicity of the title compound in HCT116 colon cancer cells was mediated through apoptotic cell death.
To obtain chemotherapeutic agents that induce apoptosis, 21 diethyl 2,6-dimethyl- 1,4-dihydropyridine-3,5-dicarboxylates were designed and synthesized. NMR spectroscopy, ab initio calculations, and X-ray crystallography demonstrated that the diethyl 2,6-dimethyl-1,4- dihydropyridine-3,5-dicarboxylate derivatives synthesized in this research had symmetric structures. Even if the half-maximal cell growth inhibitory concentrations of the 21 derivatives did not show dramatic inhibitory activity against HCT116 human colon cancer cells, small changes in the structure affected the anticancer activities. Treatment with diethyl 4-(4-chlorophenyl)-2,6- dimethyl-1,4-dihydropyridine-3,5-dicarboxylate substantially reduced the cell viability and the cytotoxicity against HCT116 colon cancer cells was mediated through apoptotic cell death. As the ability of diethyl 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylates to induce apoptosis has not been previously reported, we have now reported their design, synthesis, cytotoxicity, and structureactivity relationships.
汉茨希酯，即2,6 - 二甲基 - 1,4 - 二氢吡啶 - 3,5 - 二羧酸二乙酯，已被用作氢供体，并且其多种生物学效应已有报道。为了鉴定具有凋亡作用的化疗药物，设计并合成了21种2,6 - 二甲基 - 1,4 - 二氢吡啶 - 3,5 - 二羧酸二乙酯；迄今为止，它们尚未作为凋亡诱导剂被报道。研究了它们的结构 - 细胞毒性关系。对标题化合物进行了进一步的生物学实验。
使用克隆形成试验在HCT116人结肠癌细胞中测量当前合成化合物的细胞毒性。采用膜联蛋白V染色试验来确认标题化合物是否诱导凋亡。为了鉴定合成化合物，进行了核磁共振（NMR）光谱和高分辨率质谱（HR - MS）分析。由于在NMR光谱数据中观察到分子对称性，通过从头算和X射线晶体学确定了三维结构。
从NMR光谱、从头算和X射线晶体学获得的结果表明，本研究中合成的2,6 - 二甲基 - 1,4 - 二氢吡啶 - 3,5 - 二羧酸二乙酯衍生物具有对称结构。在HCT116人结肠癌细胞系中测试了21种衍生物的细胞毒性，其半数最大细胞生长抑制浓度在16.29至68.88 μM之间。结构 - 细胞毒性关系表明，较大的取代基更受青睐，对位取代基往往具有更好的细胞毒性值，并且极性可能也起作用。标题化合物在HCT116结肠癌细胞中的细胞毒性是通过凋亡性细胞死亡介导的。
为了获得诱导凋亡的化疗药物，设计并合成了21种2,6 - 二甲基 - 1,4 - 二氢吡啶 - 3,5 - 二羧酸二乙酯。NMR光谱、从头算和X射线晶体学表明，本研究中合成的2,6 - 二甲基 - 1,4 - 二氢吡啶 - 3,5 - 二羧酸二乙酯衍生物具有对称结构。即使21种衍生物的半数最大细胞生长抑制浓度对HCT116人结肠癌细胞没有显示出显著的抑制活性，但结构上的微小变化会影响抗癌活性。用4 -（4 - 氯苯基）- 2,6 - 二甲基 - 1,4 - 二氢吡啶 - 3,5 - 二羧酸二乙酯处理可显著降低细胞活力，并且对HCT116结肠癌细胞的细胞毒性是通过凋亡性细胞死亡介导的。由于2,6 - 二甲基 - 1,4 - 二氢吡啶 - 3,5 - 二羧酸二乙酯诱导凋亡的能力此前尚未见报道，我们现在报道了它们的设计、合成、细胞毒性和构效关系。
