Cellular uptake of glycoside-based transporters

细胞摄取糖苷转运蛋白

• 批准号: 7266070
• 负责人: YITZHAK TOR
• 金额: $ 26万
• 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/7266070
• 关键词: Aminoglycosides; Ammonium; Binding; Biological Assay; Carbamates; Cell Line; Cell membrane; Cell surface; Cells; Characteristics; Charge; Chemicals; Chinese Hamster Ovary Cell; Classification; Clinic; Development; Diffusion; Disease; Drug Delivery Systems; Drug Kinetics; 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; guanidinium; in vivo; interdisciplinary approach; mutant; neoplastic cell; novel; novel therapeutics; programs; receptor; scaffold; size; 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）：这个项目的长期目标是提高我们对配体-细胞表面相互作用的认识，并将其应用于新的细胞递送剂的设计、合成和实现。为此，我们通过衍生氨基糖苷合成了一系列胍基糖苷转运体，并表明各种细胞系通过含有聚糖硫酸肝素的带负电荷的细胞表面蛋白聚糖有效地结合和吸收这些化合物。胍苷类充当转运体，增强细胞对其他不可渗透分子的吸收。这些特定的目标为设计和合成新的胍苷类苷、评估它们在不同细胞系中的细胞摄取和定位以及评估它们递送具有治疗潜力的物质的能力提供了一个全面的方法。合成系统修饰的胍苷类并评价其细胞摄取。为了全面了解总体电荷的影响，以及胍基的分布和三维投影，将合成一系列衍生物。为了确定最有效的递送载体，将利用基于细胞的结合和摄取测定建立结构-活性关系。目标2。评估不同细胞类型中胍苷类的结合和摄取。将研究胍苷苷在人、鼠肿瘤细胞和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。目标3。胍苷糖苷类药物偶联物的合成及细胞摄取评价。最有前途的含胍衍生物将被偶联到分子货物（包括小分子和高分子量蛋白质）上，并将评估它们的细胞摄取情况。我们将研究毒素的传递是否可以用来阻止肿瘤生长，以及溶酶体储存缺陷是否可以通过转运体介导的酶替代来纠正。基于机制的转导支架的开发将识别先导结构并促进有用药物递送载体的开发。新的分子递送载体将扩大可用的工具库，以控制治疗药物的定位和释放。因此，拟议的工作旨在获取有关新转运蛋白的信息，并为治疗疾病提供一个平台。