DNA-linked dendrimer nanoparticle systems for diagnosis

用于诊断的 DNA 连接树枝状聚合物纳米颗粒系统

• 批准号: 7050850
• 负责人: JAMES R. BAKER
• 金额: $ 50万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-29 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7050850
• 关键词: SCID mouse; atomic force microscopy; calorimetry; chelating agents; chemical conjugate; chemical synthesis; circular dichroism; cisplatin; complementary DNA; cytotoxicity; diagnosis design /evaluation; doxorubicin; fluorescent dye /probe; folate; gadolinium; high performance liquid chromatography; mass spectrometry; methotrexate; nanotechnology; neoplasm /cancer chemotherapy; neoplasm /cancer diagnosis; nonhuman therapy evaluation; nuclear magnetic resonance spectroscopy; oligonucleotides; paclitaxel; pharmacokinetics; polymers; surface plasmon resonance; therapy design /development

DESCRIPTION (provided by applicant): Targeted dendrimer-based nanodevices have shown excellent promise in both in vitro cell culture and in vivo animal studies as cancer therapeutics. However, each device must be custom synthesized for a particular set of targeting molecules, imaging agents, and desired therapeutics. We propose a unique solution to this limitation by developing single function dendrimer modules linked by complementary oligonucleotides. This allows targeting, imaging, and therapeutic dendrimers to be combined into multifunctional therapeutics simply by heating mixtures of these agents above the annealing temperature of the oligonucleotide duplex. The project will consist of five specific aims. Specific Aim 1 will involve the design and synthesis of complementary oligonucleotides conjugated to poly(amido)amine dendrimers or dendrons to achieve the desired structural topologies. The ability to construct complex devices will be assessed using well-defined targeting molecules (folic acid, her2 antibodies and RGD peptides), drugs (methotrexate, Taxol, cis-platin and doxorubicin), imaging agents (Gadolinium chelators and fluorescent dyes). Specific Aim 2 will characterize the self-assembled nanodevices using techniques including PAGE, HPLC, CE, Mass Spectroscopy, NMR, AFM, and NSOM. Specific Aim 3 involves testing the DNA-linked nanodevices for binding and internalization in vitro; the avidity and specificity of binding will be examined using CD, differential calorimetry and Biacore analyses. Devices carrying therapeutics will be tested for effectiveness at inducing cell death, and all devices will also be tested for inherent cytotoxicity. Specific Aim 4 employs animal models to assess the effectiveness of the dendrimer linked therapeutics to treat tumors in vivo. In addition, the biodistribution of the therapeutics will be assessed using radiolabeled material and a novel fiber optic probe that uses two-photon excitation with femto-second pulses. Finally, under Specific Aim 5 we will work with the NCI nanoparticle characterization lab in Frederick to make the materials developed in this program available to other investigators. This platform has the potential to revolutionize cancer therapeutics and facilitate "personalized medicine." Lay description: We are designing a method and the tools for developing targeted cancer drugs that can be tailored to the needs of an individual patient. The physician can select various components and the components are then linked together like "tinker toys" to make a personalized medicine. This medicine would selectively target only the cancer, thereby avoiding the nausea, hair loss and illness caused by regular cancer chemotherapy.

描述（由申请人提供）： 基于树枝状大分子的靶向纳米器件在体外细胞培养和体内动物研究中作为癌症治疗剂显示出了良好的前景。然而，每个设备必须针对一组特定的靶向分子、显像剂和所需的治疗进行定制合成。我们通过开发由互补寡核苷酸连接的单功能树枝状聚合物模块，提出了针对这一限制的独特解决方案。这使得只需将这些试剂的混合物加热到寡核苷酸双链体的退火温度以上，就可以将靶向、成像和治疗性树枝状聚合物组合成多功能治疗剂。该项目将包括五个具体目标。具体目标 1 将涉及设计和合成与聚（酰胺）胺树枝状聚合物或树枝状分子缀合的互补寡核苷酸，以实现所需的结构拓扑。将使用明确的靶向分子（叶酸、her2 抗体和 RGD 肽）、药物（甲氨蝶呤、紫杉醇、顺铂和阿霉素）、显像剂（钆螯合剂和荧光染料）来评估构建复杂装置的能力。具体目标 2 将使用 PAGE、HPLC、CE、质谱、NMR、AFM 和 NSOM 等技术来表征自组装纳米器件。具体目标 3 涉及测试 DNA 连接纳米器件的体外结合和内化；将使用 CD、差示量热法和 Biacore 分析来检查结合的亲合力和特异性。携带治疗药物的设备将接受诱导细胞死亡的有效性测试，所有设备也将接受固有的细胞毒性测试。具体目标 4 采用动物模型来评估树枝状大分子连接疗法在体内治疗肿瘤的有效性。此外，将使用放射性标记材料和使用飞秒脉冲双光子激发的新型光纤探针来评估治疗药物的生物分布。最后，根据特定目标 5，我们将与弗雷德里克的 NCI 纳米颗粒表征实验室合作，将该计划中开发的材料提供给其他研究人员。该平台有可能彻底改变癌症治疗方法并促进“个性化医疗”。 简单描述：我们正在设计一种方法和工具，用于开发可根据个体患者的需求定制的靶向癌症药物。医生可以选择各种组件，然后将这些组件像“修补玩具”一样连接在一起，制成个性化药物。这种药物将选择性地仅针对癌症，从而避免常规癌症化疗引起的恶心、脱发和疾病。