Cellular uptake of glycoside-based transporters

细胞摄取糖苷转运蛋白

• 批准号: 7919905
• 负责人: YITZHAK TOR
• 金额: $ 5.36万
• 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 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7919905
• 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）：该计划的长期目标是提高我们对配体-细胞表面相互作用的认识，并将其应用于新细胞递送剂的设计、合成和实施。为此，我们已经合成了一系列的胍基糖苷转运衍生氨基糖苷类，并已表明，各种细胞系的结合，并采取有效地通过负电荷的细胞表面蛋白聚糖含有聚糖，硫酸乙酰肝素的化合物。胍基糖苷类作为转运蛋白，增强细胞对其他不可渗透分子的摄取。具体的目标提供了一个全面的方法设计和合成新的胍基糖苷类，评估其细胞摄取和定位在不同的细胞系，并评估其能力，提供货物的治疗潜力：AIM 1。合成系统修饰的胍基糖苷并评估其细胞摄取。为了全面了解整体电荷的影响，以及胍基团的分布和三维投影，将合成一系列衍生物。将使用基于细胞的结合和摄取测定开发结构-活性-关系，目的是鉴定最有效的递送载体。AIM 2.评价胍基糖苷类在不同细胞类型中的结合和摄取。将研究胍基糖苷类在人和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。AIM 3.合成并评估胍基糖苷类药物偶联物的细胞摄取。最有前途的含胍衍生物将与分子货物（包括小分子和高分子量蛋白质）结合，并评价其细胞摄取。我们将研究是否可以利用毒素的传递来阻断肿瘤生长，以及是否可以通过转运蛋白介导的酶替代来纠正溶酶体储存缺陷。基于机制的开发转导支架将确定铅结构，并促进有用的药物输送车辆的发展。新的分子传递载体将扩大可用于控制治疗剂的定位和释放的工具库。因此，拟议的工作旨在获得有关新转运蛋白的信息，并提供治疗疾病的平台。