Using Dendrimers to Design Multivalent Therapeutic Agents

使用树枝状聚合物设计多价治疗剂

• 批准号: 8720009
• 负责人: Mary J Cloninger
• 金额: $ 29.3万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8720009
• 关键词: Acetylgalactosamine; Acquired Immunodeficiency Syndrome; Adhesions; Affinity; Alzheimer' s Disease; Animal Model; Antigens; Arbitration; Architecture; Avidity; Binding; Biological; Biological Assay; Biological Models; Biological Process; Biology; Caenorhabditis elegans; Carbohydrates; Cell Adhesion; Cell physiology; Cells; Chemotaxis; Collaborations; Dendrimers; Development; Diabetes Mellitus; Disease; Environment; Enzyme-Linked Immunosorbent Assay; Event; Exhibits; Exposure to; Fluorescence; Fluorescence Anisotropy; Funding; Galactose; Galactose Binding Lectin; Galectin 1; Galectin 3; Generations; Goals; Individual; Influenza; Institutes; Kinetics; Lactose; Ligands; Malignant Neoplasms; Mediating; Mediator of activation protein; Metalloproteases; Migration Assay; Molecular Sieve Chromatography; N-acetyllactosamine; Neoplasm Metastasis; Pathway interactions; Pattern; Peptides; Play; Process; Prodrugs; Property; Proteins; Relative (related person); Research; Role; Structure; Surface Plasmon Resonance; System; Techniques; Testing; Therapeutic; Therapeutic Agents; Thompson-Friedenreich Antigen; Toxic effect; Universities; angiogenesis; base; cancer cell; carbohydrate binding protein; design; extracellular; in vivo; light scattering; molecular recognition; mouse model; novel; programs; receptor; research study; small molecule; tool

DESCRIPTION (provided by applicant): The overarching goal of this research program is to develop synthetic multivalent frameworks for the discernment and manipulation of biological recognition processes that are important in intercellular interactions. Multivalency plays a critical role in many biological interactions, and the development of synthetic multivalent systems to systematically probe these processes is critical for advancement of the rational development of multivalent therapeutics. Because typical individual receptor/ligand interactions in biology are often weak, augmentation of these interactions using multivalency can enhance functional avidity. Moreover, multivalent interactions can create patterns of ligands that can be used to modulate processes in novel ways, and these architectures of structure cannot be formed by traditional small-molecule therapeutics. The proposed research describes the development of dendrimers as tools to advance the understanding of the requirements for multivalent frameworks that are designed to mediate multivalent cancer cellular recognition processes. Our hypothesis is that carbohydrate functionalized dendrimers can be used to form clusters of galectins and that these galectin/glycodendrimer clusters will effectively arbitrate the intercellular recognition events of cancer cells. We have chosen glycodendrimers as the multivalent frameworks for the proposed studies because of the ease of manipulation of their size (generation) and of their end group functionalization. We have chosen galectins as our target proteins because of their known functions in cancer processes. The three specific aims are as follows. 1) Synthesize new carbohydrate-functionalized dendrimers. 2) Characterize the glycodendrimer/galectin binding interactions. 3) Perform cell based and animal model (C. elegans) assays with glycodendrimers. For specific aim 1, chemoenzymatic syntheses to form N-acetyllactosamine and Thomsen-Friedenriech antigen functionalized dendrimers are proposed. The synthesis of dendrimers bearing peptide substrates for matrix metalloproteases is also proposed. For specific aim 2, characterization of glycodendrimer/galectin aggregates is proposed using fluorescence lifetime waveform, fluorescence anisotropy, size exclusion chromatography-multiangle light scattering, surface plasmon resonance and ELISA experiments. Because of the effect of galectins -1 and -3 on cancer cellular aggregation processes, our goal is to characterize the binding affinity of the glycodendrimer/galectin interactions and the pattern of galectin that is displayed to the cells in the presence of glycodendrimers. For specific aim 3, we will perform angiogenesis and cellular adhesion assays in collaboration with Dr. Avraham Raz of the Karmanos Cancer Institute at Wayne State University. We also propose to perform homotypic cellular aggregation assays, chemotaxis and migration assays, mechanoelastic studies, kinetic studies on release of prodrug mimics, and studies with C. elegans.

描述(由申请人提供)：该研究计划的总体目标是开发合成的多价框架，用于识别和操纵在细胞间相互作用中重要的生物识别过程。多价性在许多生物相互作用中起着至关重要的作用，开发合成的多价系统来系统地研究这些过程对于促进多价疗法的合理发展至关重要。由于生物学中典型的单个受体/配体的相互作用通常很弱，使用多价态来增强这些相互作用可以增强功能亲和力。此外，多价相互作用可以产生配体的模式，可以用新的方式调节过程，而这些结构的结构不能通过传统的小分子疗法形成。这项拟议的研究描述了树枝状大分子作为工具的开发，以促进对多价框架的需求的理解，这些框架旨在中介多价癌细胞识别过程。我们的假设是，碳水化合物功能化的树状大分子可以用来形成Galectins簇，并且这些Galectin/糖树状大分子簇将有效地仲裁癌细胞的细胞间识别事件。我们选择糖树状大分子作为多价骨架，因为它们的大小(生成)和端基官能化很容易操纵。我们选择Galectins作为我们的目标蛋白，是因为它们在癌症过程中的已知功能。三个具体目标如下。1)合成新型碳水化合物功能化树枝状大分子。2)表征糖树状大分子与半乳糖凝集素的结合作用。3)用糖树枝状大分子进行细胞和动物模型(线虫)检测。针对具体目标1，提出了化学酶法合成N-乙酰乳糖胺和Thomsen-Friedenriech抗原功能化树枝状大分子。还提出了合成含基质金属蛋白酶多肽底物的树枝状大分子。针对具体目标2，提出了利用荧光寿命波形、荧光各向异性、尺寸排除层析-多角度光散射、表面等离子体共振和ELISA法来表征糖树枝状大分子/半乳糖凝集素聚集体。由于Galectins-1和Galectins-3对癌细胞聚集过程的影响，我们的目标是表征糖树状大分子/Galectin相互作用的结合亲和力，以及在糖树状大分子存在的情况下Galectin向细胞显示的模式。对于特定的目标3，我们将与韦恩州立大学卡马诺斯癌症研究所的Avraham Raz博士合作进行血管生成和细胞黏附分析。我们还建议进行同型细胞聚集分析、趋化和迁移分析、机械弹性研究、前药模拟物释放动力学研究以及对线虫的研究。