Department of Safety Engineering, Dongguk University Wise, 123 Dongdae-ro, Gyeongju-si 780714, Gyeongbuk, Republic of Korea.
Molecules. 2023 Aug 8;28(16):5942. doi: 10.3390/molecules28165942.
The present study describes the synthesis, characterization, and in vitro molecular interactions of a steroid 3β,6β-diacetoxy-5α-cholestan-5-ol. Through conventional and solid-state methods, a cholestane derivative was successfully synthesized, and a variety of analytical techniques were employed to confirm its identity, including high-resolution mass spectrometry (HRMS), Fourier transforms infrared (FT-IR), nuclear magnetic resonance (NMR), elemental analysis, and X-ray single-crystal diffraction. Optimizing the geometry of the steroid was undertaken using density functional theory (DFT), and the results showed great concordance with the data from the experiments. Fluorescence spectral methods and ultraviolet-vis absorption titration were employed to study the in vitro molecular interaction of the steroid regarding human serum albumin (HSA). The Stern-Volmer, modified Stern-Volmer, and thermodynamic parameters' findings showed that steroids had a significant binding affinity to HSA and were further investigated by molecular docking studies to understand the participation of active amino acids in forming non-bonding interactions with steroids. Fluorescence studies have shown that compound interacts with human serum albumin (HSA) through a static quenching mechanism. The binding affinity of compound for HSA was found to be 3.18 × 10 mol, and the Gibbs free energy change (ΔG) for the binding reaction was -9.86 kcal mol at 298 K. This indicates that the binding of compound to HSA is thermodynamically favorable. The thermodynamic parameters as well as the binding score obtained from molecular docking at various Sudlow's sites was -8.2, -8.5, and -8.6 kcal/mol for Sites I, II, and III, respectively, supporting the system's spontaneity. Aside from its structural properties, the steroid demonstrated noteworthy antioxidant activity, as evidenced by its IC value of 58.5 μM, which is comparable to that of ascorbic acid. The findings presented here contribute to a better understanding of the pharmacodynamics of steroids.
本研究描述了一种甾体 3β,6β-二乙酰氧基-5α-胆甾烷-5-醇的合成、表征和体外分子相互作用。通过常规和固态方法，成功合成了一种胆甾烷衍生物，并采用高分辨率质谱 (HRMS)、傅里叶变换红外 (FT-IR)、核磁共振 (NMR)、元素分析和 X 射线单晶衍射等多种分析技术来确认其结构。通过密度泛函理论 (DFT) 对甾体的几何形状进行了优化，结果与实验数据非常吻合。荧光光谱法和紫外可见吸收滴定法用于研究甾体与人血清白蛋白 (HSA) 的体外分子相互作用。Stern-Volmer、修正 Stern-Volmer 和热力学参数的结果表明，甾体对 HSA 具有显著的结合亲和力，并通过分子对接研究进一步研究了它们与 HSA 形成非键相互作用的活性氨基酸的参与。荧光研究表明，化合物 通过静态猝灭机制与人血清白蛋白 (HSA) 相互作用。发现化合物 与 HSA 的结合亲和力为 3.18×10 mol，在 298 K 时，结合反应的吉布斯自由能变化 (ΔG) 为-9.86 kcal mol。这表明化合物 与 HSA 的结合在热力学上是有利的。从分子对接在不同 Sudlow 部位获得的热力学参数和结合评分分别为-8.2、-8.5 和-8.6 kcal/mol，支持系统的自发性。除了其结构特性外，该甾体还表现出显著的抗氧化活性，其 IC 值为 58.5 μM，与抗坏血酸相当。本研究结果有助于更好地了解甾体的药效动力学。
