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: 8236550
• 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 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8236550
• 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. PUBLIC HEALTH RELEVANCE: Multivalent interactions, which occur when two or more molecular recognition events take place simultaneously between two partners, play a critical role in many biological processes. Diseases with multivalent aspects include AIDS, influenza, Alzheimer's disease, diabetes, and cancer. The proposed research describes the development of glycodendrimers as tools to advance the understanding of the requirements for multivalent frameworks that are designed to mediate multivalent cancer cellular recognition processes. The hypothesis that will be tested in the proposed research is that glycodendrimers will form clusters of galectins (carbohydrate binding proteins) and that these galectin/glycodendrimer clusters will effectively arbitrate intercellular recognition events.

描述（由申请人提供）：该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重要的生物识别过程。多价在许多生物相互作用中起着至关重要的作用，开发合成多价系统来系统地探测这些过程对于促进多价治疗的合理发展至关重要。由于生物学中典型的个体受体/配体相互作用通常很弱，因此使用多价增强这些相互作用可以增强功能亲和力。此外，多价相互作用可以创建配体模式，可用于以新颖的方式调节过程，而这些结构体系无法通过传统的小分子疗法形成。 拟议的研究描述了树枝状聚合物的开发作为工具，以促进对旨在介导多价癌症细胞识别过程的多价框架的要求的理解。我们的假设是碳水化合物功能化的树枝状聚合物可用于形成半乳糖凝集素簇，并且这些半乳糖凝集素/糖树枝状聚合物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拟议研究的多价框架，因为它们的大小（生成）和端基功能化易于操作。我们选择半乳糖素作为我们的靶蛋白，因为它们在癌症过程中具有已知的功能。 这三个具体目标如下。 1）合成新的碳水化合物功能化树枝状聚合物。 2) 表征糖树枝状聚合物/半乳糖凝集素结合相互作用。 3) 使用糖树枝状聚合物进行基于细胞和动物模型（线虫）的检测。 对于具体目标 1，提出了化学酶合成法以形成 N-乙酰基乳糖胺和 Thomsen-Friedenriech 抗原功能化树枝状聚合物。还提出了带有基质金属蛋白酶肽底物的树枝状聚合物的合成。对于具体目标 2，建议使用荧光寿命波形、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 实验来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以及在糖树枝状聚合物存在下向细胞展示的半乳糖凝集素模式。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测定、趋化性和迁移测定、机械弹性研究、前药模拟物释放的动力学研究以及线虫研究。 公共卫生相关性：多价相互作用是当两个或多个分子识别事件在两个伙伴之间同时发生时发生的，在许多生物过程中发挥着关键作用。具有多价方面的疾病包括艾滋病、流感、阿尔茨海默病、糖尿病和癌症。拟议的研究描述了糖树枝状聚合物的开发作为工具，以促进对旨在介导多价癌症细胞识别过程的多价框架的要求的理解。在拟议的研究中将测试的假设是，糖树枝状聚合物将形成半乳糖凝集素（碳水化合物结合蛋白）簇，并且这些半乳糖凝集素/糖树枝状聚合物簇将有效地仲裁细胞间识别事件。