Although a plethora of targeted anticancer small molecule drugs became available, the low response rate and drug resistance imply the continuous need for expanding the anticancer chemical space. In this study, a novel series of nicotinonitrile derivatives was designed, synthesized and evaluated for cytotoxic activities in HepG2 and MCF-7 cells. All derivatives showed high to moderate cytotoxic activity against both cell lines, with cell-type and chemotype-dependent cytotoxic potential. The normal HEK-293 T cells were ca. 50-fold less susceptible to the cytotoxic effect of the inhibitors. The in vitro enzyme inhibitory activity of selected active cytotoxic derivatives 8c, 8e, 9a, 9e and 12 showed that they have sub- to one digit micromolar 50 % inhibitory concentration (IC₅₀) against the three Pim kinase isoforms, with 8e being the most potent (IC₅₀ ≤ 0.28 μM against three Pim kinases), comparable to the pan kinase inhibitor, Staurosporine. In HepG2, 8e induced cell cycle arrest at the G2/M phase. Apoptotic mechanistic studies with 8c and 8e in HepG2 cells, indicated a significant upregulation in both P53 and caspase-3 relative gene expression, as well as increased Bax/Bcl-2 protein expression level. Further, docking studies combined with molecular dynamic simulation showed a stable complex with high binding affinity of 8e to Pim-1 kinase; exploiting a negative electrostatic potential surface interaction with the added dimethyl amino group in the new compounds. Moreover, in silico ADME profile prediction indicated that all compounds are orally bioavailable and most of them can penetrate the blood–brain barrier. This study presents novel nicotinonitrile derivatives as auspicious hits for further optimization as antiproliferative agents against liver cancer cells and promising pan Pim kinase inhibitors at submicromolar concentrations.

尽管出现了大量靶向抗癌小分子药物，但低反应率和耐药性意味着需要不断扩大抗癌化学空间。在本研究中，设计、合成了一系列新型烟腈衍生物，并评估了其在 HepG2 和 MCF-7 细胞中的细胞毒活性。所有衍生物对两种细胞系都显示出高到中度的细胞毒活性，具有细胞类型和化学类型依赖性细胞毒潜力。正常的 HEK-293 T 细胞是 ca。对抑制剂的细胞毒性作用的敏感性降低 50 倍。选定活性细胞毒衍生物8c , 8e , 9a , 9e和12的体外酶抑制活性表明它们对三种 Pim 激酶亚型具有亚至一位数微摩尔的 50% 抑制浓度 (IC _{50 )，其中}8e是最有效的（针对三种 Pim 激酶的 IC ₅₀  ≤ 0.28 μM），与泛激酶抑制剂相当，星形孢菌素。在 HepG2 中，8e诱导细胞周期停滞在 G2/M 期。在 HepG2 细胞中使用8c和8e进行的细胞凋亡机制研究表明 P53 和 caspase-3 相关基因表达显着上调，并且 Bax/Bcl-2 蛋白表达水平增加。此外，对接研究与分子动力学模拟相结合显示出具有8e高结合亲和力的稳定复合物Pim-1 激酶；利用与新化合物中添加的二甲基氨基的负静电势表面相互作用。此外，计算机ADME 概况预测表明所有化合物都具有口服生物利用度，并且其中大部分可以穿透血脑屏障。本研究将新型烟腈衍生物作为进一步优化的吉祥物，作为抗肝癌细胞的抗增殖剂和亚微摩尔浓度的有前途的泛 Pim 激酶抑制剂。