Ismail Magda M F, Shawer Taghreed Z, Ibrahim Rabab S, Elnagar Mohamed R, Ammar Yousry A
Department of Pharmaceutical Chemistry, Faculty of Pharmacy (Girls), Al-Azhar University, Cairo 11754 Egypt.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy (Girls), Al-Azhar University, Cairo 11754 Egypt.
Bioorg Chem. 2025 Mar;156:108182. doi: 10.1016/j.bioorg.2025.108182. Epub 2025 Jan 20.
The vascular endothelial growth factor receptor is essential for the angiogenesis of cancer. Tumor propagation was effectively suppressed by inhibiting VEGFR-2 activity. As a result, the target quinoxaline-pyrazole hybrids were created in a way that closely resembled the structural characteristics of VEGFR-2 inhibitors. The synthesized compounds were assessed in vitro using the MTT assay with doxorubicin serving as a reference standard for their cytotoxic properties against the HCT-116 and MCF-7 cell lines. Additionally, when tested on human normal fibroblasts (WI38), the promising cytotoxic compounds 8, 11, 13, and 15 were shown to be selective to cancer cells. Using ELISA, they showed mechanistically inhibitory activities against VEGFR-2; compound 13 was the most effective inhibitor, with an IC of 0.045 ± 6.24 uM, surpassing sorafenib (IC of 0.049 ± 5.24 μM). Notably, it was discovered that our target compound, 13, was 1.1 times more potent than sorafenib and 3.19 times more potent than sunitinib as a VGFRA2 inhibitor. Furthermore, Western blot analysis revealed that its VEGFR2 protein levels were noticeably higher than the control. Compound 13's selectivity towards VEGFR2 was further confirmed by testing it against other kinases, such as PDGFRA (IC 0.329 ± 0.014 μM) and EGFR (IC 0.6 ± 0.019 μM). Furthermore, 13 demonstrated a 50 % decrease in VEGF-A secretion in comparison to the control group, demonstrating its anti-angiogenic quality. A scratch closure percentage of 57.78 %, which was much lower than the 97.04 percent of untreated control cells, showed 13's anti-migratory capability. According to the cell cycle study, compound 13 induces apoptosis at the sub-G1 phase and terminates the cell cycle at the G1 phase. Consequently,flow cytometric analysis revealed that it caused apoptosis; compound 13 increases the BAX/Bcl-2 ratio from control to 13.66, and it also activates caspase 3 to 422.48 ± 43.82 and induces p53 to 366.79 ± 40.21. Docking simulations revealed potential binding modes and crucial structural elements of active drugs, and they were almost in agreement with enzymatic examination. For every hybrid, in silico physicochemical attributes, drug likeness metrics, and ligand efficiency were plausible. It's interesting to note that 13 and 15 are plausible medication candidates.
血管内皮生长因子受体对癌症血管生成至关重要。通过抑制VEGFR - 2活性可有效抑制肿瘤增殖。因此，以与VEGFR - 2抑制剂结构特征极为相似的方式合成了目标喹喔啉 - 吡唑杂化物。使用MTT法在体外评估合成的化合物，以阿霉素作为针对HCT - 116和MCF - 7细胞系细胞毒性特性的参考标准。此外，在人正常成纤维细胞（WI38）上进行测试时，有前景的细胞毒性化合物8、11、13和15对癌细胞具有选择性。通过ELISA检测，它们对VEGFR - 2显示出机制性抑制活性；化合物13是最有效的抑制剂，IC50为0.045±6.24 μM，超过了索拉非尼（IC50为0.049±5.24 μM）。值得注意的是，发现我们的目标化合物13作为VGFRA2抑制剂比索拉非尼强1.1倍，比舒尼替尼强3.19倍。此外，蛋白质印迹分析显示其VEGFR2蛋白水平明显高于对照组。通过针对其他激酶如PDGFRA（IC50 0.329±0.014 μM）和EGFR（IC50 0.6±0.019 μM）进行测试，进一步证实了化合物13对VEGFR2的选择性。此外，与对照组相比，13显示VEGF - A分泌减少了50%，表明其具有抗血管生成特性。划痕闭合率为57.78%，远低于未处理对照细胞的97.04%，显示出13的抗迁移能力。根据细胞周期研究，化合物13在亚G1期诱导细胞凋亡，并在G1期终止细胞周期。因此，流式细胞术分析显示其诱导了细胞凋亡；化合物13使BAX/Bcl - 2比值从对照增加到13.66，还将caspase 3激活至422.48±43.82并诱导p53至366.79±40.21。对接模拟揭示了活性药物的潜在结合模式和关键结构元件，并且它们与酶学检测结果几乎一致。对于每个杂化物，计算机模拟的物理化学属性、类药性质指标和配体效率都是合理的。有趣的是，13和15是合理的药物候选物。
