Chemistry of Natural Compounds Department, National Research Centre, Dokki, 12622 Giza, Egypt.
Biochemistry Department, Faculty of Agriculture, Cairo University, 12613 Giza, Egypt.
Molecules. 2020 Mar 3;25(5):1124. doi: 10.3390/molecules25051124.
The present work aims to design and synthesize novel series of spiro pyrazole-3,3'-oxindoles analogues and investigate their bioactivity as antioxidant and antimicrobial agents, as well as antiproliferative potency against selected human cancerous cell lines (i.e., breast, MCF-7; colon, HCT-116 and liver, HepG-2) relative to healthy noncancerous control skin fibroblast cells (BJ-1). The mechanism of their cytotoxic activity has been also examined by immunoassaying the levels of key anti- and proapoptotic protein markers. The analytical and spectral data of the all synthesized target congeners were compatible with their structures. Synthesized compounds showed diverse moderate to powerful antimicrobial and antioxidant activities. Results of MTT assay revealed that seven synthesized compounds (i.e., 11a, 11b, 12a, 12b, 13b, 13c and 13h) particularly exhibited significant cytotoxicity against the three cancerous cell lines under investigation. Ranges of IC values obtained were 5.7-21.3 and 5.8-37.4 µg/mL against HCT-116 and MCF-7, respectively; which is 3.8 and 6.5-fold (based on the least IC values) more significant relative to the reference chemotherapeutic drug doxorubicin. In HepG-2 cells, the analogue 13h the highest cytotoxicity with IC value of 19.2µg/mL relative to doxorubicin (IC = 21.6µg/mL). The observed cytotoxicity was specific to cancerous cells, as evidenced by the minimal toxicity in the noncancerous control skin-fibroblast cells. ELISA results indicated that the observed antiproliferative effect against examined cancer cell lines is mediated engaging the activation of apoptosis as illustrated by the significant increase in proapoptotic protein markers (p53, bax and caspase-3) and reduction in the antiapoptotic marker bcl-2. Taken together, results of the present study emphasize the potential of spiro pyrazole-oxindole analogues as valuable candidate anticancer agents against human cancer cells.
本工作旨在设计和合成一系列新型螺吡唑-3,3'-氧吲哚类似物，并研究它们作为抗氧化剂和抗菌剂的生物活性，以及相对于健康的非癌细胞皮肤成纤维细胞 (BJ-1)，对选定的人类癌细胞系（即乳腺癌 MCF-7；结肠癌 HCT-116 和肝癌 HepG-2）的抗增殖活性。还通过免疫测定关键抗凋亡和促凋亡蛋白标志物的水平来检查它们细胞毒性活性的机制。所有合成目标同系物的分析和光谱数据与它们的结构一致。合成化合物表现出不同程度的中等至强的抗菌和抗氧化活性。MTT 测定结果表明，七种合成化合物（即 11a、11b、12a、12b、13b、13c 和 13h）特别对三种受研究的癌细胞系表现出显著的细胞毒性。获得的 IC 值范围分别为 5.7-21.3 和 5.8-37.4µg/mL，与参考化疗药物阿霉素相比，分别高出 3.8 和 6.5 倍（基于最低的 IC 值）。在 HepG-2 细胞中，类似物 13h 表现出最高的细胞毒性，IC 值为 19.2µg/mL，相对于阿霉素（IC = 21.6µg/mL）。观察到的细胞毒性是针对癌细胞的，这可以从非癌细胞皮肤成纤维细胞中的最小毒性得到证明。ELISA 结果表明，观察到的对受检癌细胞系的增殖抑制作用是通过激活凋亡来介导的，如图所示，促凋亡蛋白标志物（p53、bax 和 caspase-3）显著增加，抗凋亡标志物 bcl-2 减少。总之，本研究结果强调了螺吡唑-氧吲哚类似物作为有价值的候选抗癌剂对抗人类癌细胞的潜力。
