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）：该项目的长期目标是增进我们对配体-细胞表面相互作用的了解，并将其应用于新细胞递送剂的设计、合成和实施。为此，我们通过氨基糖苷类衍生化合成了一系列胍基糖苷转运蛋白，并表明各种细胞系通过含有聚糖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。胍基糖苷充当转运蛋白，增强细胞对其他不可渗透分子的摄取。具体目标为设计和合成新的胍基糖苷、评估其在不同细胞系中的细胞摄取和定位以及评估其递送具有治疗潜力的货物的能力提供了综合方法：AIM 1. 合成系统修饰的胍基糖苷并评估他们的细胞摄取。为了全面了解总电荷的影响以及胍基的分布和三维投影，将合成一系列衍生物。将使用基于细胞的结合和摄取测定来开发结构-活性-关系，目的是确定最有效的递送载体。目标 2. 评估不同细胞类型中胍基糖苷的结合和摄取。将研究胍基糖苷在人类和小鼠肿瘤细胞以及人类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。目标 3. 合成并评估胍基糖苷-药物缀合物的细胞摄取。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，并评估它们的细胞摄取。我们将研究是否可以利用毒素的传递来阻止肿瘤生长，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。基于机制的转导支架的开发将识别先导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展可用于控制治疗药物的定位和释放的工具库。因此，拟议的工作旨在获取有关新转运蛋白的信息并提供治疗疾病的平台。