Farouk Faten, Sharaky Marwa, Elkady Ehab F, Sayed Rawda M
Pharmaceutical Chemistry Department, Faculty of Pharmacy, Ahram Canadian University, 6th of October, Giza 12451, Egypt.
Pharmacology Unit, Cancer Biology Department, National Cancer Institute Cairo University, Kasr El-Aini St., Cairo 11562, Egypt.
J Chromatogr B Analyt Technol Biomed Life Sci. 2025 Mar 1;1253:124466. doi: 10.1016/j.jchromb.2025.124466. Epub 2025 Jan 13.
Glucocorticoids (GCs) are hallmarks of anti-inflammatory activity. They are used as adjuvant therapy in oncology medications to alleviate some of the associated side effects. Although recent research has indicated that GCs have favorable anticancer potential, some scientific evidence suggests a pro-proliferation impact of GCs on cancer cells. This may create a scientific confusion on the utility of GCs and the choice of which GC enhances the anticancer potential and mitigates the negative effect. Accurate in-silico prediction and correct ranking of biological activity may be limited by the impact of the physicochemical interaction between small molecules and biological membranes. Chromatographic retention is inherently dependent on the physicochemical properties of the test molecule. It can scale the relative hydrophobicity and tentatively indicate the membrane permeability. In this study, the relationship between the in-silico binding affinity, the chromatographic retention and the in-vitro anticancer activity was investigated. Fifteen GCs were chromatographically separated on an Inertsil® C18 (4.6 * 250 mm; 5 μm) HPLC column. The binding affinity of the test GCs was determined on three receptors involved in cancer cell proliferation (topoisomerase II (TOPII), glucocorticoid receptor (GR) and ATP-binding cassette (ABCG2)). The antiproliferative potential of the test steroids was determined as per their IC. The correlation between chromatographic retention and the binding affinity to the observed IC was investigated by multiple linear regression (MLR). Results revealed that some GCs exhibited a remarkably favorable inhibitory potential against cancer cell lines over normal cell lines. Our data indicated a significant correlation between the retention times of different GCs and the determined binding affinity, especially to the GR (r = 0.677; p = 0.011) and the estrone sulphate (ESS) binding site of the ABCG2 (r = 0.643; p = 0.018). Concurrently, the retention times were well correlated to the observed IC on the colorectal cancer cell lines (r = 0.580; p = 0.038) and the hypopharyngeal cell lines (r = 0.638; p = 0.019). Significant MLR models (n = 4) correlating the retention times of the tested steroids and the binding affinity to the observed IC were created. The MLR model correlating the retention times and the ABCG2_ESS binding affinity to the IC on lung cancer was the most significant (p = 0.004). The accuracy of the model was 107.12 ± 29.18 % indicating good IC prediction abilities. In conclusion, chromatographic retention can be employed as a low-cost and simple auxiliary tool for improving the in-silico prediction of the in-vitro activities of small molecules.
糖皮质激素（GCs）是抗炎活性的标志。它们在肿瘤药物中用作辅助治疗，以减轻一些相关的副作用。尽管最近的研究表明GCs具有良好的抗癌潜力，但一些科学证据表明GCs对癌细胞有促增殖作用。这可能会在GCs的效用以及选择哪种GC增强抗癌潜力并减轻负面影响方面造成科学上的困惑。小分子与生物膜之间的物理化学相互作用的影响可能会限制生物活性的准确计算机模拟预测和正确排序。色谱保留本质上取决于测试分子的物理化学性质。它可以衡量相对疏水性并初步表明膜通透性。在本研究中，研究了计算机模拟结合亲和力、色谱保留与体外抗癌活性之间的关系。在Inertsil® C18（4.6 * 250 mm；5μm）HPLC柱上对15种GCs进行了色谱分离。在参与癌细胞增殖的三种受体（拓扑异构酶II（TOPII）、糖皮质激素受体（GR）和ATP结合盒（ABCG2））上测定了测试GCs的结合亲和力。根据其IC50测定了测试类固醇的抗增殖潜力。通过多元线性回归（MLR）研究了色谱保留与对观察到的IC50的结合亲和力之间的相关性。结果显示，一些GCs对癌细胞系的抑制潜力明显优于正常细胞系。我们的数据表明，不同GCs的保留时间与测定的结合亲和力之间存在显著相关性，尤其是与GR（r = 0.677；p = 0.011）和ABCG2的硫酸雌酮（ESS）结合位点（r = 0.643；p = 0.018）。同时，保留时间与在结肠癌细胞系（r = 0.580；p = 0.038）和下咽细胞系（r = 0.638；p = 0.019）上观察到的IC50密切相关。建立了将测试类固醇的保留时间与对观察到的IC50的结合亲和力相关联的显著MLR模型（n = 4）。将保留时间和ABCG2_ESS结合亲和力与肺癌IC50相关联的MLR模型最为显著（p = 0.004）。该模型的准确率为107.12±29.18%，表明具有良好的IC50预测能力。总之，色谱保留可作为一种低成本且简单的辅助工具，用于改进小分子体外活性的计算机模拟预测。
