Cellular uptake of glycoside-based transporters

描述(由申请人提供)：该项目的长期目标是提高我们对配体-细胞表面相互作用的知识，并将其应用于新的细胞递送剂的设计、合成和实施。为此，我们通过对氨基糖苷类化合物的衍生化合成了一系列的胍基糖苷转运蛋白，并证明了不同的细胞系通过含有硫酸乙酰肝素的带负电荷的细胞表面蛋白多糖有效地结合和吸收这些化合物。胍基糖苷作为转运体，增强细胞对其他不透性分子的摄取。这些特定的目标为设计和合成新的胍糖苷提供了一种综合的方法，评估了它们在不同细胞系中的细胞摄取和定位，并评价了它们输送治疗潜力的能力：目的1.合成系统修饰的胍糖苷并评价它们的细胞摄取。为了全面了解总电荷的影响，以及胍基的分布和三维投影，我们合成了一系列的衍生物。将利用基于细胞的结合和摄取分析建立结构-活性-关系，目的是确定最有效的递送载体。目的2.评价不同细胞类型对鸟嘌呤糖苷的结合和摄取。将研究鸟嘌呤糖苷在人和小鼠肿瘤细胞以及人成纤维细胞中的结合和摄取，并与CHO细胞中的机制进行比较。目的3.合成并评价鸟嘌呤糖苷-药物结合物的细胞摄取。最有希望的含鸟苷衍生物将被连接到分子货物(包括小分子和高相对分子蛋白质)，并将评估它们的细胞摄取。我们将检查是否可以利用毒素的传递来阻止肿瘤的生长，以及是否可以通过转运蛋白介导的酶替代来纠正溶酶体存储缺陷。基于机制的转导支架的开发将识别铅结构，并促进有用的药物输送载体的开发。新的分子递送载体将扩大可用于控制治疗药物本地化和释放的工具的保留范围。因此，拟议的工作旨在获取有关新的转运体的信息，并为治疗疾病提供一个平台。

DESCRIPTION (provided by applicant): The long-term goal of this program is to advance our knowledge of ligand-cell surface interactions and apply it to the design, synthesis and implementation of new cellular delivery agents. Towards this end, we have synthesized a series of guanidinoglycoside transporters by derivatizing aminoglycosides and have shown that various cell lines bind and take up the compounds efficiently via negatively-charged cell-surface proteoglycans containing the glycan, heparan sulfate. The guanidinoglycosides act as transporters, enhancing the cellular uptake of otherwise impermeable molecules. The specific aims provide a comprehensive approach for the design and synthesis of new guanidinoglycosides, assessment of their cellular uptake and localization in different cell lines, and their evaluation of their ability to deliver cargos of therapeutic potential: AIM 1. Synthesize systematically modified guanidinoglycosides and evaluate their cellular uptake. To understand comprehensively the impact of the overall charge, as well as the distribution and three- dimensional projection of guanidinium groups, a series of derivatives will be synthesized. A structure-activity-relationship will be developed using cell-based assays for binding and uptake with the objective of identifying the most effective delivery vehicle. AIM 2. Evaluate binding and uptake of guanidinoglycosides in different cell types. The binding and uptake of guanidinoglycosides in human and murine tumor cells and in human fibroblasts will be investigated and compared to the mechanisms in CHO cells. AIM 3. Synthesize and evaluate cellular uptake of guanidinoglycoside-drug conjugates. The most promising guanidinium-containing derivatives will be conjugated to molecular cargos (including small molecules and high MW proteins) and their cellular uptake will be evaluated. We will examine if delivery of toxins can be exploited to block tumor growth and if lysosomal storage deficiency can be corrected by enzyme replacement mediated via transporters. A mechanism-based development of transduction scaffolds will identify lead structures and facilitate the development of useful drug delivery vehicles. New molecular delivery vehicles will expand the repertoire of tools available to control the localization and release of therapeutics. The proposed work thus aims to acquire information about new transporters and provide a platform for treating disease.