Ghafoor Naeem Abdul, Yildiz Aysegul
Department of Molecular Biology and Genetics, Graduate School of Natural and Applied Sciences, Mugla Sitki Kocman University, 48000 Mugla, Turkey.
Department of Molecular Biology and Genetics, Faculty of Science, Mugla Sitki Kocman University, 48000 Mugla, Turkey.
ACS Omega. 2023 Sep 15;8(38):34583-34596. doi: 10.1021/acsomega.3c03471. eCollection 2023 Sep 26.
Cancer remains a major cause of morbidity and mortality worldwide, and while current therapies, such as chemotherapy, immunotherapy, and cell therapy, have been effective in many patients, the development of novel therapeutic options remains an urgent priority. Mouse double minute 2 (MDM2) is a key regulator of the tumor suppressor protein p53, which plays a critical role in regulating cellular growth, apoptosis, and DNA repair. Consequently, MDM2 has been the subject of extensive research aimed at developing novel cancer therapies. In this study, we employed a machine learning-based approach to establish a quantitative structure-activity relationship model capable of predicting the potential in vitro efficacy of small molecules as MDM2 inhibitors. Our model was used to screen 5883 FDA-approved drugs, resulting in the identification of promising hits that were subsequently evaluated using molecular docking and molecular dynamics simulations. Two antihistamine drugs, cetirizine (CZ) and rupatadine (RP), exhibited particularly favorable results in the initial in silico analyses. To further assess their potential use as the activators of the p53 pathway, we investigated the antiproliferative capability of the abovementioned drugs on human glioblastoma and neuroblastoma cell lines. Both the compounds exhibited significant antiproliferative effects on the abovementioned cell lines in a dose-dependent manner. The half-maximal inhibitory concentration (IC) of CZ was found to be 697.87 and 941.37 μM on U87 and SH-SY5Y cell lines, respectively, while the IC of RP was found to be 524.28 and 617.07 μM on the same cell lines, respectively. Further investigation by quantitative reverse transcriptase polymerase chain reaction analysis revealed that the CZ-treated cell lines upregulate the expression of the p53-regulated genes involved in cell cycle arrest, apoptosis, and DNA damage response compared to their respective vehicle controls. These findings suggest that CZ activates the p53 pathway by inhibiting MDM2. Our results provide compelling preclinical evidence supporting the potential use of CZ as a modulator of the MDM2-p53 axis and its plausible repurposing for cancer treatment.
癌症仍然是全球发病和死亡的主要原因，虽然目前的疗法，如化疗、免疫疗法和细胞疗法，在许多患者中都很有效，但开发新的治疗选择仍然是当务之急。小鼠双微体2（MDM2）是肿瘤抑制蛋白p53的关键调节因子，p53在调节细胞生长、凋亡和DNA修复中起关键作用。因此，MDM2一直是旨在开发新型癌症疗法的广泛研究的主题。在本研究中，我们采用基于机器学习的方法建立了一个定量构效关系模型，该模型能够预测小分子作为MDM2抑制剂的潜在体外疗效。我们的模型用于筛选5883种FDA批准的药物，从而确定了有前景的命中物，随后使用分子对接和分子动力学模拟对其进行评估。两种抗组胺药物，西替利嗪（CZ）和卢帕他定（RP），在最初的计算机模拟分析中表现出特别有利的结果。为了进一步评估它们作为p53通路激活剂的潜在用途，我们研究了上述药物对人胶质母细胞瘤和神经母细胞瘤细胞系的抗增殖能力。这两种化合物均以剂量依赖性方式对上述细胞系表现出显著的抗增殖作用。发现CZ在U87和SH-SY5Y细胞系上的半数最大抑制浓度（IC）分别为697.87和941.37μM，而RP在相同细胞系上的IC分别为524.28和617.07μM。通过定量逆转录聚合酶链反应分析进一步研究发现，与各自的溶剂对照相比，CZ处理的细胞系上调了参与细胞周期停滞、凋亡和DNA损伤反应的p53调节基因的表达。这些发现表明CZ通过抑制MDM2激活p53通路。我们的结果提供了令人信服的临床前证据，支持CZ作为MDM2-p53轴调节剂的潜在用途及其在癌症治疗中的合理重新利用。
