Proteomic Studies of Dendrimer-based Nanomedicines

基于树枝状聚合物的纳米药物的蛋白质组学研究

• 批准号: 8022918
• 负责人: W. Andy Tao
• 金额: $ 18.31万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8022918
• 关键词: Apoptosis; Area; Biodistribution; Biological Markers; Biological Models; Biomedical Research; Categories; Cell Extracts; Cells; Charge; Data; Dendrimers; Development; Disease; Drug Carriers; Drug Delivery Systems; Drug Design; Event; Exposure to; Gene Delivery; General Population; Generations; Goals; Image; In Vitro; Isotope Labeling; Knowledge; Life; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Medicine; Methods; Molecular; Molecular Target; Organic Synthesis; Organism; Outcome; Pharmaceutical Preparations; Phase; Phosphoric Monoester Hydrolases; Preparation; Prodrugs; Property; Protein Tyrosine Phosphatase; Proteins; Proteome; Proteomics; Public Health; Reagent; Reporting; Research; Research Personnel; Role; Route; Sensitivity and Specificity; Series; Solid; Specificity; Staging; Surface; System; Techniques; Technology; Therapeutic; Therapeutic Agents; Tissues; Water; base; biomaterial compatibility; cancer cell; clinical Diagnosis; design; drug candidate; drug discovery; functional group; improved; in vivo; innovation; member; nano; nanomaterials; nanomedicine; nanoparticle; novel; novel therapeutics; phosphatase inhibitor; prototype; public health relevance; response; scaffold; small molecule; therapeutic target; tool

DESCRIPTION (provided by applicant): Despite numerous reports of the design and delivery of novel nanomedicines such as dendrimer-drug conjugates, only relatively few studies have examined their therapeutic potential at the molecular level. In particular, there are currently no convenient and quick approaches to identify specific subsets of intracellular targets of drug candidates. The long-term goals of this proposal are to apply proteomic approaches to identify intracellular drug targets and to develop novel therapeutic agents based on nanoparticles. The objective of this application is to use dendrimer-protein tyrosine phosphatase (PTP) inhibitor conjugates as a model system to establish a proteomic platform that efficiently and reproducibly characterizes the specificity and activity of these nanomedicines. Our rationale is that dendrimers functionalized with PTP inhibitors will not only efficiently deliver these small drug candidates intracellularly, but will also provide a powerful proteomic tool to probe their therapeutic targets in living cells. The proposed research will capitalize on our ability to create multi-functional groups on the surface of dendrimers such that they can achieve efficient intracellular delivery for specific targeting and then can be readily isolated for subsequent proteomic analyses. Guided by strong preliminary data, this much needed technology will be achieved by pursuing three specific aims: 1) to synthesize multi-functionalized dendrimers containing an immobilized PTP inhibitor; 2) to identify PTP targets in cancer cell homogenates; and 3) to identify PTP protein targets in intact cancer cells in culture. Under aim 1, an already proven synthetic route will be applied to the preparation of multifunctionalized dendrimer agents. Under aims 2 and 3, the activity and specificity of these dendrimer agents will be examined in vitro and in living cells, respectively. This project is innovative as the proposed studies will be among the first to combine the use of dendrimers as drug carriers (nanomedicines) and potential therapeutic agents with proteomics to identify therapeutic targets. It will capitalize on a powerful technique to directly identify therapeutic targets in living cells. Once the utility of this strategy is established, it is highly likely that it can be expanded to the characterization of a large number of nanomedicines based on dendrimers and other nanoparticles. The fully developed tool will provide detailed knowledge at the molecular level on precisely how nanomedicines react and interact with their intracellular targets. The research will be of significance because it expands the important role of proteomics and its contribution to a new field and provides a powerful tool needed for drug discovery that will allow us to evaluate the actions of nanomedicines at the molecular level. In addition, functionalized dendrimers and other related nanoparticles will provide unique materials for use as novel proteomics reagents. PUBLIC HEALTH RELEVANCE: This application is proposing noteworthy studies that have the potential applicability to identifying therapeutic targets in living systems in high throughput, as well as to developing new generation medicine, in particular nanomedicine. The proposed research has strong relevance to general public health, since the technology can be potentially expanded to any disease and drug discovery area.

描述（由申请人提供）：尽管有许多关于新型纳米药物（如树枝状聚合物-药物缀合物）的设计和递送的报道，但只有相对较少的研究在分子水平上考察了它们的治疗潜力。特别地，目前没有方便和快速的方法来鉴定候选药物的细胞内靶标的特定子集。该提案的长期目标是应用蛋白质组学方法来鉴定细胞内药物靶点，并开发基于纳米颗粒的新型治疗剂。本申请的目的是使用树枝状聚合物-蛋白酪氨酸磷酸酶（PTP）抑制剂缀合物作为模型系统，以建立蛋白质组学平台，该平台有效且可再现地表征这些纳米药物的特异性和活性。我们的基本原理是，PTP抑制剂功能化的树枝状聚合物将不仅有效地提供这些小的候选药物细胞内，但也将提供一个强大的蛋白质组学工具，以探测其在活细胞中的治疗靶点。拟议的研究将利用我们在树枝状聚合物表面上创建多官能团的能力，使它们可以实现有效的细胞内递送，用于特异性靶向，然后可以很容易地分离用于随后的蛋白质组学分析。在强有力的初步数据的指导下，这项急需的技术将通过追求三个具体目标来实现：1）合成含有固定化PTP抑制剂的多功能树枝状聚合物; 2）鉴定癌细胞匀浆中的PTP靶标; 3）鉴定培养物中完整癌细胞中的PTP蛋白靶标。在目标1下，已经证明的合成路线将被应用于制备多官能化树枝状聚合物试剂。根据目标2和3，将分别在体外和活细胞中检查这些树枝状聚合物试剂的活性和特异性。这个项目是创新的，因为拟议的研究将是第一个联合收割机结合使用树枝状聚合物作为药物载体（纳米医学）和潜在的治疗剂与蛋白质组学，以确定治疗靶点。它将利用强大的技术直接识别活细胞中的治疗靶点。一旦这种策略的实用性建立，它是非常有可能的，它可以扩展到大量的基于树枝状聚合物和其他纳米粒子的纳米药物的表征。这个完全开发的工具将在分子水平上提供关于纳米药物如何与其细胞内靶点反应和相互作用的详细知识。这项研究将具有重要意义，因为它扩大了蛋白质组学的重要作用及其对新领域的贡献，并为药物发现提供了一个强大的工具，使我们能够在分子水平上评估纳米药物的作用。此外，官能化的树枝状聚合物和其他相关的纳米颗粒将提供独特的材料，用作新的蛋白质组学试剂。 公共卫生相关性：该申请提出了值得注意的研究，这些研究具有潜在的适用性，可用于以高通量识别生命系统中的治疗靶点，以及开发新一代医学，特别是纳米医学。这项研究与公众健康有很强的相关性，因为这项技术可以扩展到任何疾病和药物发现领域。

项目成果

Chemical enrichment of tyrosine phosphopeptides.

酪氨酸磷酸肽的化学富集。

• 发表时间: 2011
• 期刊: Se pu = Chinese journal of chromatography
• 作者: Hu,Lianghai;Tao,WAndy
• 通讯作者: Tao,WAndy

Phosphorylation assay based on multifunctionalized soluble nanopolymer.

• DOI: 10.1021/ac2000708
• 发表时间: 2011-04-01
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Iliuk, Anton;Martinez, Juan S.;Hall, Mark C.;Tao, W. Andy
• 通讯作者: Tao, W. Andy

Aptamer in bioanalytical applications.

• DOI: 10.1021/ac201057w
• 发表时间: 2011-06-15
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Iliuk, Anton B.;Hu, Lianghai;Tao, W. Andy
• 通讯作者: Tao, W. Andy