Li Qingqing, Lai Ka Lun, Chan Pui Shan, Leung Sui Chu, Li Ho Yin, Fang Yuan, To Kenneth K W, Choi Chung Hang J, Gao Qian Ying, Lee Thomas W Y
School of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong SAR, China.
Division of Biomedical Engineering, Department of Electronic Engineering, Faculty of Engineering, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong SAR, China.
Colloids Surf B Biointerfaces. 2016 Apr 1;140:278-286. doi: 10.1016/j.colsurfb.2015.12.053. Epub 2016 Jan 2.
The objective of this study was to fabricate dasatinib-loaded nanoparticles and evaluate their efficacy in inhibiting cellular processes of the retinal pigment epithelium (RPE) related to proliferative vitreoretinopathy (PVR), for which there are no approved pharmacological approaches. We successfully encapsulated dasatinib, a poorly soluble multi-targeted tyrosine kinase inhibitor which has great potential for the treatment of PVR, into nanoparticles prepared from micellation of PEG-b-PCL. The size of the nanomicelles was approximately 55nm with a narrow distribution. They increased the solubility of dasatinib by 475× and provided a sustained drug release. ARPE-19, an immortal RPE cell line, was used to assess the in vitro efficacy of micellar dasatinib because the RPE is believed to play a key role in the pathogenesis of PVR. Three cell-based assays, namely, proliferation, adhesion and migration, which represent three important PVR-related cellular changes of the RPE, were conducted and the cytotoxicity of micelles was also evaluated. Both blank and dasatinib-loaded micelles were non-cytotoxic towards ARPE-19 cells. Micellar dasatinib significantly inhibited cell proliferation, adhesion and migration compared to the free drug; this might be attributable to enhanced solubility. PEG-b-PCL micelles were taken up into the ARPE-19 cells by an energy-dependent clatharin and caveolae-mediated endocytosis. Our results indicated that cellular uptake and the anti-proliferation effect of drugloaded micelles were linearly correlated. Drug loading appears to be a critical parameter for cellular uptake which in turn impacts the in vitro bioactivities of polymeric micelles. Our results clearly demonstrated that dasatinib-encapsulated micelles offer considerable promise in the management of PVR.
本研究的目的是制备载有达沙替尼的纳米颗粒，并评估其在抑制视网膜色素上皮（RPE）与增殖性玻璃体视网膜病变（PVR）相关的细胞过程中的功效，目前尚无批准的治疗PVR的药理学方法。我们成功地将达沙替尼（一种具有治疗PVR巨大潜力的难溶性多靶点酪氨酸激酶抑制剂）封装到由PEG-b-PCL胶束化制备的纳米颗粒中。纳米胶束的尺寸约为55nm，分布狭窄。它们使达沙替尼的溶解度提高了475倍，并实现了药物的持续释放。ARPE-19是一种永生的RPE细胞系，由于RPE被认为在PVR的发病机制中起关键作用，因此被用于评估胶束化达沙替尼的体外功效。进行了三种基于细胞的试验，即增殖、黏附和迁移试验，它们代表了RPE与PVR相关的三种重要细胞变化，并评估了胶束的细胞毒性。空白胶束和载有达沙替尼的胶束对ARPE-19细胞均无细胞毒性。与游离药物相比，胶束化达沙替尼显著抑制细胞增殖、黏附和迁移；这可能归因于溶解度的提高。PEG-b-PCL胶束通过能量依赖的网格蛋白和小窝介导的内吞作用被ARPE-19细胞摄取。我们的结果表明，载药胶束的细胞摄取与抗增殖作用呈线性相关。载药量似乎是细胞摄取的关键参数，进而影响聚合物胶束的体外生物活性。我们的结果清楚地表明，封装达沙替尼的胶束在PVR的治疗中具有很大的前景。
