Borik Rita M, Hussein Mohammed A
Department of Physical Sciences, Chemistry Division, College of Science, Jazan University, P.O. Box. 114, Jazan 45142, Kingdom of Saudi Arabia.
Biotechnology Department, Faculty of Applied Health Science Technology, October 6 University, Giza 28125, Egypt.
Curr Pharm Des. 2025;31(12):957-980. doi: 10.2174/0113816128337881241016064641.
PGK1 and PKM2 are glycolytic enzymes, and their expression is upregulated in cancer cells. STAT3 is a transcription factor implicated in breast cancer progression and chemoresistance. Researchers worldwide continue to explore how targeting genes might lead to more effective anti-breast cancer therapies. The present study aims to synthesize quinazolines containing caffeoyl moiety for developing innovative anticancer agents against the human breast cancer cell line (MCF-7).
A new quinazoline 2 was synthesized by reacting caffeic acid with 5-amino-phenylpyrazole carboxylate 1 in the presence of PCl3. Compound 2 reacted with NHNH.HO to produce compound 3 through cyclo-condensation. Apoptosis and necrosis as well as inhibition activity compounds 2 and 3 against PGK1, and PKM2 were evaluated. The effect of compounds 2 and 3 on the levels of GSH, GR, SOD, GPx, CAT, MDA, Bax, Bcl-2, caspase-3, P53 and VEGF levels as well as PGK1, PKM2 and STAT3 gene expression were estimated in MCF-7 tumor cells.
The viability of MCF-7 cells was reduced to 22.42% and 45.86% after incubation with compounds 2 and 3 for 48 hours, respectively. The IC values for compounds 2 and 3 are 62.05 μg/mL and 16.73 μg/mL. Furthermore, compound 3 exhibited more significant apoptosis and necrosis than compound 2. IC values of compound 2 against PGK1, and PKM2 at interval concentration equals 1.04, and 0.32 μM/mL, respectively, after 210 minutes of incubation. Moreover, compound 3 were revealed strong inhibition of PGK1, and PKM2 with IC values equals 0.55 and 0.21 μg/mL, respectively after 210 minutes of incubation. Our results proved that the incubation of compounds 2 and 3 with MCF-7 cells increased the levels of apoptotic proteins, elevated MDA, Bax, caspase-3 and P53 levels, depleted GSH, GR, SOD, GPx, CAT, Bcl-2 levels and downregulated the levels of STAT3, PGK1, and PKM2 gene expression significantly. Our results proved that compound 2 showed a stronger estimated binding affinity with a ΔG of -7.2, -7.5, and -7.9 kcal/mol., respectively towards PGK1, PKM2 and STAT3 proteins. Also, compound 3 exhibits a strong binding affinity with ΔG of -7.9, -8.5, and - 8.7 kcal/mol., towards PGK1, PKM2 and STAT3 proteins.
The results show that compounds 2 and 3 induce apoptotic activity by blocking the PGK1- PKM2-STAT3 signaling pathway. The present investigation opens exciting possibilities for developing innovative new anticancer quinazolines bearing caffeoyl moiety.
PGK1和PKM2是糖酵解酶，它们在癌细胞中的表达上调。STAT3是一种与乳腺癌进展和化疗耐药性相关的转录因子。世界各地的研究人员继续探索靶向基因如何能带来更有效的抗乳腺癌治疗方法。本研究旨在合成含有咖啡酰部分的喹唑啉，以开发针对人乳腺癌细胞系（MCF-7）的创新抗癌药物。
在三氯化磷存在下，咖啡酸与5-氨基-苯基吡唑羧酸酯1反应合成了一种新的喹唑啉2。化合物2与NHNH.HO反应通过环缩合生成化合物3。评估了化合物2和3的凋亡和坏死情况以及对PGK1和PKM2的抑制活性。估计了化合物2和3对MCF-7肿瘤细胞中谷胱甘肽（GSH）、谷胱甘肽还原酶（GR）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPx）、过氧化氢酶（CAT）、丙二醛（MDA）、Bax、Bcl-2、半胱天冬酶-3、P53和血管内皮生长因子（VEGF）水平以及PGK1、PKM2和STAT3基因表达的影响。
与化合物2和3孵育48小时后，MCF-7细胞的活力分别降至22.42%和45.86%。化合物2和3的半数抑制浓度（IC）值分别为62.05μg/mL和16.73μg/mL。此外，化合物3比化合物2表现出更显著的凋亡和坏死。孵育210分钟后，化合物2对PGK1和PKM2的间隔浓度IC值分别等于1.04和0.32μM/mL。此外，孵育210分钟后，化合物3对PGK1和PKM2表现出强烈抑制作用，IC值分别为0.55和0.21μg/mL。我们的结果证明，化合物2和3与MCF-7细胞孵育会增加凋亡蛋白水平，升高MDA、Bax、半胱天冬酶-3和P53水平，消耗GSH、GR、SOD、GPx、CAT、Bcl-2水平，并显著下调STAT3、PGK1和PKM2基因表达水平。我们的结果证明，化合物2对PGK1、PKM2和STAT3蛋白分别显示出更强的估计结合亲和力，其自由能变化（ΔG）分别为-7.2、-7.5和-7.9kcal/mol。此外，化合物3对PGK,1、PKM2和STAT3蛋白表现出强烈的结合亲和力，ΔG分别为-7.9、-8.5和-8.7kcal/mol。
结果表明，化合物2和3通过阻断PGK1-PKM2-STAT3信号通路诱导凋亡活性。本研究为开发带有咖啡酰部分的创新新型抗癌喹唑啉开辟了令人兴奋的可能性。
