Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad, India.
J Biomol Struct Dyn. 2020 Mar;38(5):1539-1550. doi: 10.1080/07391102.2019.1614481. Epub 2019 May 14.
In this study, forskolin-loaded human serum albumin nanoparticles (FR-HSANPs) were successfully prepared by incorporation and affinity-binding methods. FR-HSANPs were characterized by transmission electron microscope that most of them are circular in shape and size is around 340 nm. The drug loading was more than 88% and further sustained release profiles were observed as it is 77.5% in 24 h time. Additionally, the cytotoxicity results with HepG2 cells indicated that FR-HSANPs showed significantly higher cytotoxicity and lower cell viability as compared to free forskolin (FR). Furthermore, to understand the binding mechanism of human serum albumin (HSA) with forskolin resulted from fluorescence quenching as a static mechanism and the binding constant is 6.26 ± 0.1 × 10 M, indicating a strong binding affinity. Further, association and dissociation kinetics of forskolin-HSA was calculated from surface plasmon resonance spectroscopy and the binding constant found to be = 3.4 ± 0.24 × 10 M and also fast dissociation was observed. Further, we used circular dichroism and molecular dynamics simulations to elucidate the possible structural changes including local conformational changes and rigidity of the residues of both HSA and HSA-forskolin complexes.Communicated by Ramaswamy H. Sarma.
在这项研究中，通过包合和亲和结合的方法成功制备了福司可林负载的人血清白蛋白纳米粒（FR-HSANPs）。通过透射电子显微镜对 FR-HSANPs 进行了表征，发现它们大多数呈圆形，粒径约为 340nm。载药量超过 88%，并观察到进一步的持续释放曲线，24 小时时达到 77.5%。此外，用 HepG2 细胞进行的细胞毒性结果表明，与游离福司可林（FR）相比，FR-HSANPs 表现出更高的细胞毒性和更低的细胞活力。此外，通过荧光猝灭作为静态机制，理解人血清白蛋白（HSA）与福司可林结合的机制，得出结合常数为 6.26±0.1×10M，表明具有很强的结合亲和力。进一步，从表面等离子体共振光谱计算了福司可林-HSA 的缔合和解离动力学，发现结合常数为 = 3.4±0.24×10M，并且观察到快速解离。此外，我们使用圆二色性和分子动力学模拟来阐明可能的结构变化，包括 HSA 和 HSA-福司可林复合物中残基的局部构象变化和刚性。由 Ramaswamy H. Sarma 交流。
