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: 8916768
• 负责人: Mary J Cloninger
• 金额: $ 30.21万
• 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 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8916768
• 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）：本研究计划的总体目标是开发合成多价框架，用于识别和操纵在细胞间相互作用中很重要的生物识别过程。多价在许多生物相互作用中起着关键作用，并且开发合成的多价系统以系统地探测这些过程对于推进多价治疗剂的合理开发至关重要。由于生物学中典型的个体受体/配体相互作用通常较弱，因此使用多价性增强这些相互作用可以增强功能性亲合力。此外，多价相互作用可以产生配体的模式，这些配体可以用于以新的方式调节过程，并且这些结构的架构不能由传统的小分子治疗剂形成。 拟议的研究描述了树枝状聚合物作为工具的发展，以促进对多价框架的要求的理解，这些框架旨在介导多价癌症细胞识别过程。我们的假设是，碳水化合物官能化的树枝状聚合物可用于形成半乳糖凝集素簇，这些半乳糖凝集素/glycodendrimer簇将有效地仲裁癌细胞的细胞间识别事件。我们选择糖树枝状聚合物作为多价框架的建议的研究，因为它们的大小（代）和它们的端基官能化的操作容易。我们选择半乳糖凝集素作为我们的靶蛋白，因为它们在癌症过程中的已知功能。 这三个具体目标如下。1)合成新的碳水化合物官能化的树枝状大分子。2)表征糖树枝状聚合物/半乳糖凝集素结合相互作用。3)进行基于细胞和动物模型（C. elegans）测定。 对于具体目标1，提出了形成N-乙酰基乳糖胺和B-F抗原官能化树枝状聚合物的化学酶促合成。还提出了合成树枝状大分子轴承基质金属蛋白酶的肽底物。对于具体目标2，提出使用荧光寿命波形、荧光各向异性、尺寸排阻色谱-多角度光散射、表面等离子体共振和ELISA实验表征糖树枝状聚合物/半乳糖凝集素聚集体。由于半乳糖凝集素-1和-3对癌细胞聚集过程的影响，我们的目标是表征糖树枝状聚合物/半乳糖凝集素相互作用的结合亲和力和在糖树枝状聚合物存在下向细胞展示的半乳糖凝集素的模式。对于具体目标3，我们将与韦恩州立大学Karmanos癌症研究所的Avraham Raz博士合作进行血管生成和细胞粘附测定。我们还建议进行同型细胞聚集试验，趋化性和迁移试验，机械弹性研究，前药模拟物释放的动力学研究，以及用C。优雅的