Li Rong, Hao Jon, Fujiwara Hideji, Xu Miao, Yang Shu, Dai Sheng, Long Yan, Swaroop Manju, Li Changhui, Vu Mylinh, Marugan Juan J, Ory Daniel S, Zheng Wei
1 National Center for Advancing Translational Sciences, National Institutes of Health , Bethesda, Maryland.
2 Poochon Scientific , Frederick, Maryland.
Assay Drug Dev Technol. 2017 May/Jun;15(4):154-166. doi: 10.1089/adt.2017.774.
Methyl-β-cyclodextrin (MβCD) reduces lysosomal cholesterol accumulation in Niemann-Pick disease type C1 (NPC1) patient fibroblasts. However, the pharmacological activity of MβCD reported by different laboratories varies. To determine the potential causes of this variation, we analyzed the mass spectrum characteristics, pharmacological activity of three preparations of MβCDs, and the protein expression profiles of NPC1 patient fibroblasts after treatment with different sources of MβCDs. Our data revealed varied mass spectrum profiles and pharmacological activities on the reduction of lysosomal cholesterol accumulation in NPC1 fibroblasts for these three preparations of MβCDs obtained from different batches and different sources. Furthermore, a proteomic analysis showed the differences of these three MβCD preparations on amelioration of dysregulated protein expression levels in NPC1 cells. The results demonstrate the importance of prescreening of different cyclodextrin preparations before use as a therapeutic agent. A combination of mass spectrum analysis, measurement of pharmacological activity, and proteomic profiling provides an effective analytical procedure for characterization of cyclodextrins for therapeutic applications.
甲基-β-环糊精（MβCD）可减少1型尼曼-皮克病（NPC1）患者成纤维细胞中的溶酶体胆固醇积累。然而，不同实验室报道的MβCD药理活性有所不同。为了确定这种差异的潜在原因，我们分析了三种MβCD制剂的质谱特征、药理活性，以及用不同来源的MβCD处理后NPC1患者成纤维细胞的蛋白质表达谱。我们的数据显示，这三种从不同批次和不同来源获得的MβCD制剂，在减少NPC1成纤维细胞溶酶体胆固醇积累方面，具有不同的质谱图谱和药理活性。此外，蛋白质组学分析表明，这三种MβCD制剂在改善NPC1细胞中失调的蛋白质表达水平方面存在差异。结果表明，在将不同环糊精制剂用作治疗剂之前进行预筛选很重要。质谱分析、药理活性测量和蛋白质组学分析相结合，为表征用于治疗应用的环糊精提供了一种有效的分析方法。
