The epidermal growth factor receptor (EGFR) enzyme plays a critical role in governing the cell cycle, positioning it as a promising target for the development of anticancer drugs. In this study, we endeavored to design and synthesize innovative EGFR inhibitors with potential applications in anticancer therapy. A novel series of compounds, namely 3-(4-(4-(1,3,4-oxadiazol-2-yl)-1H-imidazol-2-yl)phenyl)-1,2,4-oxadiazoles (30a–j), were meticulously designed using FBDD efforts and synthesized. The synthesized compounds underwent thorough characterization using ¹HNMR, ¹³CNMR, HRMS, and mass spectrum analyses. The in vitro anticancer activities of the newly developed compounds (30a–j) were evaluated against four human cancer cell lines such as prostate cancer (PC3 & DU-145), lung cancer (A549), and liver cancer (HEPG2) using the MTT method. The results, expressed as IC₅₀ values, demonstrated significant anticancer activity for several compounds, with five compounds (30a, 30b, 30c, 30i, and 30j) exhibiting superior potency compared to the established anticancer drug etoposide. Notably, compound 30a emerged as the most promising compound, displaying potent cytotoxicity. We also conducted a screening of the compounds on the normal Vero cell line, revealing a pronounced selectivity of the compounds against cancer cell lines, with no observable impact on the normal cell lines. Moreover, the synthesized compounds were investigated for their impact on enzyme EGFR activity. The findings revealed a robust inhibitory effect against the EGFR wild-type enzyme and a 10-fold inferior potency against the mutant form of EGFR. This observation underscores the potential of the new derivatives as effective EGFR^WT inhibitors with substantial anticancer efficacy. Further studies, including cell cycle analysis and apoptosis assays in HEPG2 cell lines, revealed cell cycle arrest at G1/G0 and G2 phases. We also evaluated the potential influence of compound 30a on the EGFR pathway using western blot analysis, revealing a significant inhibition of EGFR autophosphorylation in HEPG2 cells. In conclusion, our findings highlight the promise of these novel compounds as potent EGFR inhibitors, encouraging further investigation and development for the creation of novel and effective anticancer therapeutics.

中文翻译：

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3-(4-(4-(1,3,4-恶二唑-2-基)-1H-咪唑-2-基)苯基)-1,2,4-恶二唑衍生物的计算设计、合成和评估有效的表皮生长因子受体抑制剂：抗癌治疗的前瞻性策略

表皮生长因子受体（EGFR）酶在控制细胞周期中发挥着关键作用，使其成为抗癌药物开发的一个有前景的靶点。在这项研究中，我们致力于设计和合成具有抗癌治疗潜在应用的创新EGFR抑制剂。一系列新型化合物，即 3-(4-(4-(1,3,4-恶二唑-2-基)-1 H-咪唑-2-基)苯基)-1,2,4-恶二唑 ( 30a –j )，使用 FBDD 精心设计并综合。使用¹ HNMR、¹³ CNMR、HRMS 和质谱分析对合成的化合物进行了彻底的表征。使用 MTT 评估新开发的化合物 ( 30a-j ) 针对四种人类癌细胞系的体外抗癌活性，例如前列腺癌 (PC3 和 DU-145)、肺癌 (A549) 和肝癌 (HEPG2)方法。结果以 IC ₅₀值表示，表明几种化合物具有显着的抗癌活性，其中 5 种化合物（30a、30b、30c、30i和30j）与已建立的抗癌药物依托泊苷相比表现出更优异的效力。值得注意的是，化合物30a成为最有前途的化合物，显示出有效的细胞毒性。我们还在正常 Vero 细胞系上对这些化合物进行了筛选，结果表明这些化合物对癌细胞系具有显着的选择性，而对正常细胞系没有明显的影响。此外，还研究了合成的化合物对 EGFR 酶活性的影响。研究结果显示，它对 EGFR 野生型酶具有强大的抑制作用，而对 EGFR 突变型酶的抑制作用则低 10 倍。这一观察结果强调了新衍生物作为有效 EGFR ^WT抑制剂的潜力，具有显着的抗癌功效。进一步的研究，包括 HEPG2 细胞系的细胞周期分析和凋亡测定，显示细胞周期停滞在 G1/G0 和 G2 期。我们还使用蛋白质印迹分析评估了化合物30a对 EGFR 通路的潜在影响，揭示了对 HEPG2 细胞中 EGFR 自磷酸化的显着抑制。总之，我们的研究结果强调了这些新型化合物作为有效 EGFR 抑制剂的前景，鼓励进一步研究和开发，以创造新型有效的抗癌疗法。