Omar Mai H, Emam Soha H, Mikhail Demiana S, Elmeligie Salwa
Department of Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Cairo University, Cairo 11562, Egypt..
Department of Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Cairo University, Cairo 11562, Egypt.
Bioorg Chem. 2025 Jun 16;163:108691. doi: 10.1016/j.bioorg.2025.108691.
The design of pyrazole-based combretastatin A-4 analogues 4a-g depended on preservation of the cis olefinic configuration of combretastatin A-4 (CA-4) by replacing it with a pyrazole ring, aiming to enhance the cytotoxic activity. Furthermore, the pyrazoline moiety of 4a-g was included to add a polar center for binding interactions and to contribute to apoptosis induction. Synthesis of pyrazoline derivatives 4a-g was achieved through a new chalcone analogues 3a-g and were confirmed by spectral data and high-resolution mass spectroscopy. The cytotoxic activity was the intended biological target of the new compounds, and upon preliminary in vitro investigation against the NCI-60 panel at one dose, and it was found that the pyrazoline derivative 4a showed remarkable growth inhibition against fifteen cancer cell lines. Also, the cytotoxic activity of compounds 4a-g was screened in vitro against non-small cell lung cancer cell line A549 and ovarian cancer cell line OVCAR-4, along with the normal lung cell line WI-38, and their half-maximal inhibitory concentration (IC) values were calculated. Compound 4a was found to be the most potent cytotoxic agent (IC = 3.46 and 5.93 μM against OVCAR-4 and A549 cells, respectively) and the most selective derivative towards A549 cells with a selectivity index equal to 7.2. The inhibition of tubulin polymerization by the most active compound, 4a, was comparable to CA-4 and was visualized by means of immunofluorescent staining of A549 cells. Prediction of reactive oxygen species (ROS) production by pyrazole-pyrazoline hybrid 4a was confirmed as a rise of ROS level by compound 4a was observed to be almost 243 %- in respect to control- and ROS scavenger enzyme catalase inhibited apoptotic cell death induced by 4a by 10.5 %. Moreover, compound 4a caused a decline of the mitochondrial membrane potential by approximately 56 % less than the control, which confirms the presence of ROS. The apoptotic cell death by 4a was further studied, and it showed a 3.7-folds elevation of caspase-3 expression compared to the control. Additionally, a molecular docking study of compound 4a into the colchicine binding site of tubulin confirmed the anti-tubulin activity. Indeed, pyrazoline derivative 4a is a promising new druggable molecule that acts as a tubulin polymerization inhibitor and ROS-mediated apoptosis-inducing agent.
基于吡唑的康普他汀A - 4类似物4a - g的设计，是通过用吡唑环取代康普他汀A - 4（CA - 4）的顺式烯烃构型来实现的，目的是增强细胞毒性活性。此外，4a - g中引入了吡唑啉部分，以增加一个用于结合相互作用的极性中心，并有助于诱导细胞凋亡。吡唑啉衍生物4a - g是通过新的查尔酮类似物3a - g合成的，并通过光谱数据和高分辨率质谱进行了确认。细胞毒性活性是这些新化合物预期的生物学靶点，在对NCI - 60细胞系进行单剂量的初步体外研究中发现，吡唑啉衍生物4a对15种癌细胞系表现出显著的生长抑制作用。此外，还在体外对化合物4a - g针对非小细胞肺癌细胞系A549、卵巢癌细胞系OVCAR - 4以及正常肺细胞系WI - 38进行了细胞毒性活性筛选，并计算了它们的半数抑制浓度（IC）值。发现化合物4a是最有效的细胞毒性剂（对OVCAR - 4和A549细胞的IC分别为3.46和5.93 μM），并且是对A549细胞最具选择性的衍生物，选择性指数等于7.2。最具活性的化合物4a对微管蛋白聚合的抑制作用与CA - 4相当，并通过A549细胞的免疫荧光染色进行了可视化观察。通过观察到化合物4a使活性氧（ROS）水平相对于对照几乎升高243%，证实了吡唑 - 吡唑啉杂化物4a对ROS产生的预测，并且ROS清除酶过氧化氢酶抑制了4a诱导的凋亡细胞死亡的10.5%。此外，化合物4a使线粒体膜电位比对照下降了约56%，这证实了ROS的存在。进一步研究了4a诱导的凋亡细胞死亡，结果显示与对照相比，caspase - 3表达升高了3.7倍。此外，对化合物4a与微管蛋白的秋水仙碱结合位点进行的分子对接研究证实了其抗微管蛋白活性。事实上，吡唑啉衍生物4a是一种有前景的新型可成药分子，它作为微管蛋白聚合抑制剂和ROS介导的凋亡诱导剂发挥作用。
