Cellular uptake of glycoside-based transporters

细胞摄取糖苷转运蛋白

• 批准号: 7577371
• 负责人: YITZHAK TOR
• 金额: $ 25.9万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7577371
• 关键词: Aminoglycosides; Ammonium; Binding; Biological Assay; Carbamates; Cell Line; Cell membrane; Cell surface; Cells; Charge; Chemicals; Chinese Hamster Ovary Cell; Classification; Clinic; Development; Diffusion; Disease; Drug Delivery Systems; Enzymes; Evaluation; Family; Fibroblasts; Gene Silencing; Glycoside Hydrolases; Glycosides; Goals; Heparan Sulfate Proteoglycan; Heparitin Sulfate; Human; Knowledge; Lead; Life; Ligands; Lipid Bilayers; Mammalian Cell; Mediating; Membrane Transport Proteins; Molecular; Mus; Oligonucleotides; Organ; Organic Synthesis; Peptides; Peptoids; Pharmaceutical Preparations; Polysaccharides; Procedures; Proteins; Proteoglycan; Research Personnel; Series; Structure; Structure-Activity Relationship; Techniques; Therapeutic; Therapeutic Agents; Toxin; Vertebral column; Work; base; cell type; design; drug candidate; guanidinium; in vivo; interdisciplinary approach; mutant; neoplastic cell; novel; novel therapeutics; pharmacokinetic characteristic; programs; receptor; scaffold; small molecule; sulfation; tool; tumor growth; uptake

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.

描述（由申请人提供）：该程序的长期目标是提高我们对配体 - 细胞表面相互作用的了解，并将其应用于新的蜂窝递送剂的设计，综合和实施。为此，我们通过衍生化氨基糖苷的衍生化合物综合了一系列鸟苷二聚糖转运蛋白，并表明各种细胞系通过含有含有果糖的乙酰乙烷硫酸乙酰肝素硫酸盐的聚糖细胞表面蛋白聚糖有效地结合并有效地占据化合物。鸟苷聚糖充当转运蛋白，增强了原本不可渗透分子的细胞摄取。该特定目的为设计和合成新的鸟苷糖苷的设计和合成提供了一种全面的方法，评估了它们的细胞摄取和在不同细胞系中的定位，以及他们评估其提供治疗潜力的能力的能力：AIM 1。合成系统修改的鸟肾上腺素糖苷剂并评估其蜂窝状uptarlular Uptake。为了全面地了解鸟肾组的整体电荷以及分布和三维投影的影响，将合成一系列衍生物。将使用基于细胞的测定法进行结合和摄取，以识别最有效的递送工具，从而开发结构活性关系。 AIM 2。评估不同细胞类型中鸟苷二聚糖的结合和摄取。将研究人和鼠类肿瘤细胞中鸟苷糖苷以及人成纤维细胞中的结合和摄取，并将其与CHO细胞中的机制进行比较。 AIM 3。合成和评估鸟代属糖苷 - 药物结合物的细胞摄取。最有希望的含鸟嘌呤的衍生物将与分子嘉戈斯（包括小分子和高MW蛋白）结合，并评估其细胞摄取。我们将检查是否可以利用毒素的递送来阻断肿瘤的生长，以及是否可以通过通过转运蛋白介导的酶替代酶矫正溶酶体储存功能。基于机制的转导支架的发展将识别铅结构并促进有用的药物递送车的发展。新的分子输送车将扩大可用的工具库，以控制治疗剂的定位和释放。因此，拟议的工作旨在获取有关新转运蛋白的信息，并为治疗疾病提供平台。