Ayoup Mohammed Salah, Mamdouh Eman, Soliman Saied M, Elghamry Ibrahim, Nafie Mohamed S, Negm Amr, Sonousi Amr, Kassab Asmaa E, Awad Laila F
Department of Chemistry, College of Science, King Faisal University Al-Ahsa 31982 Saudi Arabia
Chemistry Department, Faculty of Science, Alexandria University P. O. Box 426 Alexandria 21321 Egypt
RSC Adv. 2025 Jun 10;15(24):19530-19545. doi: 10.1039/d5ra01428j. eCollection 2025 Jun 4.
A series of novel bis-amide Ugi adducts with a Y-shaped configuration imitating the thematic feature of fourth-generation EGFR inhibitors was designed and synthesized a one-step Ugi four-component reaction. All the synthesized Ugi adducts were evaluated for their anti-proliferative efficacy against MDA-MB-231 human breast cancer and A549 non-small cell lung cancer cell lines. Their selectivity was assessed using normal human lung epithelial cells (BEAS-2B). The Ugi adduct 5 stood up as the study hit concerning cytotoxic efficacy and selectivity. It was 1.42- and 1.20-fold more potent than 5-FU and cisplatin, respectively, against the MDA-MB-231 cell line, and 1.82- and 1.62-fold more potent than 5-FU and cisplatin, respectively, against the A549 cell line. Additionally, compound 5 demonstrated the most prominent selectivity against MDA-MB-231 and A549 cells (SI = 7 and 11.7, respectively); it was 14 to 20.9-fold safer than 5-FU and cisplatin. Accordingly, it was subjected to further enzymatic and mechanistic studies. The Ugi adduct 5 showed excellent sub-micromolar potency (IC = 0.19 μM) against human mutant EGFR enzyme equipotent with that of osimertinib (IC = 0.1 μM). It exhibited a promising IC value of 2.1 nM against the EGFR enzyme, comparable to erlotinib (IC = 1.3 nM). In comparison to the untreated control, the Ugi adduct 5 caused a decrease in the expression of the cancer initiation, angiogenic, and metastatic markers (c-Myc, CD-44, CD-133, VEGF, and TGF) to 0.36, 0.5, 0.7, 0.5, and 0.4-fold, respectively, in MDA-MB-231 cells. Regarding A549 cells, the exposure to compound 5 showed a 0.41, 0.64, 0.58, 0.71, and 0.69-fold reduction in the expression of c-Myc, CD-44, CD-133, VEGF, and TGF markers compared to the untreated control. Compound 5 markedly elevated miRNA-132 and miRNA-200c expressions in the MDA-MB-231 cell line by 3.8 and 3.1-fold, while in A549 cells, compound 5 demonstrated enhancement of miRNA-132 and miRNA-200c expression by 2.4 and 1.9-fold changes compared to that of the control. It promoted apoptosis induction caspase 3/9 activation (1.8 and 2.3-folds) in the A549 cell line. The molecular docking interpretations of the most potent Ugi adduct 5 in EGFR (PDB ID: 6LUB) and the wild-type EGFR (PDB ID: 1M17) enzymes are aligned with and explain its potential to dually inhibit EGFR and EGFR tyrosine kinases.
设计并通过一步法乌吉四组分反应合成了一系列具有Y形结构的新型双酰胺乌吉加合物，该结构模仿了第四代表皮生长因子受体（EGFR）抑制剂的主题特征。评估了所有合成的乌吉加合物对MDA-MB-231人乳腺癌细胞系和A549非小细胞肺癌细胞系的抗增殖功效。使用正常人肺上皮细胞（BEAS-2B）评估它们的选择性。就细胞毒性功效和选择性而言，乌吉加合物5表现突出。它对MDA-MB-231细胞系的效力分别比5-氟尿嘧啶（5-FU）和顺铂高1.42倍和1.20倍，对A549细胞系的效力分别比5-FU和顺铂高1.82倍和1.62倍。此外，化合物5对MDA-MB-231和A549细胞表现出最显著的选择性（选择性指数分别为7和11.7）；它比5-FU和顺铂安全14至20.9倍。因此，对其进行了进一步的酶学和作用机制研究。乌吉加合物对人突变型EGFR酶表现出优异的亚微摩尔效力（半数抑制浓度[IC]=0.19μM），与奥希替尼（IC=0.1μM）相当。它对EGFR酶的IC值为2.1 nM，与厄洛替尼（IC=1.3 nM）相当，颇具前景。与未处理的对照相比，乌吉加合物5使MDA-MB-231细胞中癌症起始、血管生成和转移标志物（c-Myc、CD-44、CD-133、血管内皮生长因子[VEGF]和转化生长因子[TGF]）的表达分别降至0.36倍、0.5倍、0.7倍、0.5倍和0.4倍。对于A549细胞，与未处理的对照相比，暴露于化合物5使c-Myc、CD-44、CD-133、VEGF和TGF标志物的表达分别降低0.41倍、0.64倍、0.58倍、0.71倍和0.69倍。化合物5使MDA-MB-231细胞系中的微小RNA-132（miRNA-132）和微小RNA-200c（miRNA-200c）表达显著升高3.8倍和3.1倍，而在A549细胞中，与对照相比，化合物5使miRNA-132和miRNA-200c表达分别提高2.4倍和1.9倍。它促进了A549细胞系中凋亡诱导——半胱天冬酶3/9激活（1.8倍和2.3倍）。最有效的乌吉加合物5在EGFR（蛋白质数据银行[PDB]编号：6LUB）和野生型EGFR（PDB编号：1M17）酶中的分子对接解释与其双重抑制EGFR和EGFR酪氨酸激酶的潜力相符，并解释了其潜力。
