Abstract：
A series of 16 novel spirooxindole analogs 8a-p were designed and constructed via cost-effective single-step multicomponent [3+2] cycloaddition reaction of azomethine ylide (AY) generated in situ from substituted isatin (6a-d) with suitable amino acids (7a-c) and ethylene-engrafted pyrazole derivatives (5a,b). The potency of all compounds was assayed against a human breast cancer cell line (MCF-7) and a human liver cell line (HepG2). Spiro compound 8c was the most active member among the synthesized candidates, with exceptional cytotoxicity against the MCF-7 and HepG2 cell lines, with IC50 values of 0.189 ± 0.01 and 1.04 ± 0.21 µM, respectively. The candidate 8c exhibited more potent activity (10.10- and 2.27-fold) than the standard drug roscovitine (IC50 = 1.91 ± 0.17 µM (MCF-7) and 2.36 ± 0.21 µM (HepG2)). Compound 8c was investigated for epidermal growth factor receptor (EGFR) inhibition; it exhibited promising IC50 values of 96.6 nM compared with 67.3 nM for erlotinib. The IC50 value of 8c (34.98 nM) exhibited cyclin-dependent kinase 2 (CDK-2) inhibition, being more active than roscovitine the (IC50 = 140 nM) in targeting the CDK-2 kinase enzyme. Additionally, for apoptosis induction of compound 8c in MCF-7, it upregulated the expression levels of proapoptotic genes for P53, Bax, caspases-3, 8, and 9 at up to 6.18, 4.8, 9.8, 4.6, 11.3 fold-change, respectively, and downregualted the level of the antiapoptotic gene for Bcl-2 by 0.14-fold. Finally, a molecular docking study of the most active compound 8c highlighted a good binding affinity with Lys89 as the key amino acid for CDK-2 inhibition.
摘要：
通过具有成本效益的单步多组分[32]环加成反应，设计并构建了一系列16种新型螺羟吲哚类似物8a-p，该反应是由取代的靛蓝（6a-d）与合适的氨基酸（7a-c）和乙烯接枝吡唑衍生物（5a，b).针对人乳腺癌细胞系(MCF-7)和人肝细胞系(HepG2)测定所有化合物的效力。螺环化合物8c是合成候选化合物中最活跃的成员，对MCF-7和HepG2细胞系具有特殊的细胞毒性，IC50值为0.189±0.01和1.04±0.21µM，分别。候选8c比标准药物roscovitine（IC50=1.91±0.17µM（MCF-7）和2.36±0.21µM（HepG2））表现出更有效的活性（10.10倍和2.27倍）。研究化合物8c的表皮生长因子受体(EGFR)抑制;与厄洛替尼的67.3nM相比,它表现出96.6nM的有希望的IC50值。8c的IC50值(34.98nM)显示细胞周期蛋白依赖性激酶2(CDK-2)抑制,在靶向CDK-2激酶方面比roscovitine更具活性（IC50=140nM）。此外,对于MCF-7中化合物8c的凋亡诱导，它上调了P53，Bax，caspases-3，8和9在高达6.18，4.8，9.8，4.6，11.3倍变化，分别,并将Bcl-2的抗凋亡基因的水平降低了0.14倍。最后,最具活性的化合物8c的分子对接研究强调了与作为CDK-2抑制的关键氨基酸的Lys89的良好结合亲和力。
