Cell-specific and multifunctional drug formulations for in vivo delivery

项目摘要 本项目的目标是：1）开发细胞特异性给药系统; 2）构建 多功能药物递送系统;和3）证明这些系统在动物中的治疗功效 用于治疗遗传疾病、传染病以及癌症的模型。体内细胞特异性递送是 药物输送领域最具挑战性的问题之一。为了解决这个问题，我们建议两个 策略：（1）模拟受体介导的病毒进入。病毒的简单结构和组成 使它们能够特异性感染人类的多种细胞类型。我们的研究计划将适用于 病毒感染的知识，以指导细胞特异性递送系统的设计。比如说， 核仁素被鉴定为人呼吸道合胞病毒在上皮细胞上的细胞受体 (RSV)在肺部。我们将利用抗核仁素抗体或核仁素配体来靶向肺上皮细胞， 细胞(2)靶向肿瘤过度表达的受体。然而，以前的研究已经确定了一些 肿瘤过度表达的受体，其中许多是相对未开发的，我们对它们的理解是 非常有限。例如，Chlorotoxin是一种短肽，能够穿过血脑屏障（BBB）， 特异性结合在脑肿瘤细胞中过度表达的基质金属蛋白酶-2（MMP-2）。 我们建议利用这些配体来特异性靶向肿瘤细胞。我们会评估他们的肿瘤 成像探针的特异性。为了开发多功能药物递送系统，我们将整合 从上述研究中鉴定的配体与不同的递送系统如纳米颗粒和缀合物。 有效负载可以是mRNA、siRNA和小分子药物，以产生多功能递送系统。的 递送系统能够利用mRNA来表达功能蛋白，利用siRNA来进行基因沉默， 以及用于癌症治疗的小分子药物。通过递送编码Cas9的mRNA和特异性引导RNA， （sgRNA），我们期望在体内实现基因工程，以找到治疗遗传性疾病的方法， 用于治疗病毒感染如B型肝炎。另外我们有 设计的氯毒素-花菁-药物（CCD）作为多功能缀合物，以特异性地递送近- 红外分子探针和细胞毒性剂用于脑转移性乳腺癌（BMBC）肿瘤细胞。后 当施用时，CCD缀合物将穿过BBB并与BMBC细胞上的MMP-2结合，触发细胞凋亡。 《防治荒漠化公约》的执行情况。花青探针将有助于成像引导的肿瘤切除。同时，细胞毒性药物 将诱导肿瘤细胞的凋亡，其低于成像的检测极限， 驱逐是不可能的。我们的研究目标是将这一研究策略的创新转化为 开发更好的药物输送工具来治疗各种疾病。

PROJECT SUMMARY The objectives of this project are 1) to develop cell-specific drug delivery systems; 2) to construct multifunctional drug delivery systems; and 3) to demonstrate therapeutic efficacy of these systems in animal models for treating genetic disorders, infectious diseases, as well as cancers. Cell-specific delivery in vivo is one of the most challenging issues in the field of drug delivery. To address this issue, we propose two strategies: (1) to mimic the receptor-mediated virus entry. The simple structure and composition of viruses make them capable of specifically infecting many cell types in humans. Our research program will apply knowledge of viral infections in order to guide the design of cell-specific delivery systems. For example, nucleolin has been identified as a cellular receptor on epithelial cells for human respiratory syncytial virus (RSV) in the lungs. We will utilize anti-nucleolin antibody or nucleolin ligands in order to target lung epithelial cells. (2) to target tumor over-expressed receptors. Previous studies, however, have identified a number of tumor over-expressed receptors, of which many are relatively unexplored and our understanding of them is very limited. For example, Chlorotoxin, a short peptide, is capable of crossing the blood brain barrier (BBB) and binding specifically to the matrix metalloproteinase-2 (MMP-2), which is over-expressed in brain tumor cells. We propose to utilize these ligands in order to specifically target tumor cells. We will evaluate their tumor specificity with imaging probes. In order to develop multifunctional drug delivery systems, we will integrate the ligands identified from the above studies with different delivery systems such as nanoparticles and conjugates. The payloads can be mRNA, siRNA, and small-molecule drugs to yield multifunctional delivery systems. The delivery systems are capable of utilizing mRNA for expressing functional proteins, siRNA for gene silencing, and small-molecule drugs for cancer therapy. By delivering mRNA encoding Cas9 and specific guide RNA (sgRNA), we anticipate achieving gene engineering in vivo in order to find a cure for genetic disorders such as hemophilia and to deplete viral genome for treating viral infections such as hepatitis B. In addition, we have designed Chlorotoxin-Cyanine-Drug (CCD) as a multifunctional conjugate in order to specifically deliver near- infrared molecular probes and cytotoxic agents to brain metastatic breast cancer (BMBC) tumor cells. After administration, CCD conjugates will cross the BBB and bind with MMP-2 on BMBC cells, triggering the cellular uptake of the CCD. Cyanine probes will facilitate imaging-guided tumor resection. Meanwhile, cytotoxic agents will induce the apoptosis of tumor cells, which are below detection limits of imaging and for which surgical removal is not a possibility. Our research goal is to translate the innovation of this research strategy in order to develop better drug delivery tools to treat diverse diseases.