Surface-modified pharmaceutical nanocarriers for subcellular targeting

描述(申请人提供)：许多药物，包括各种大分子(蛋白质、酶、抗体)，甚至载药的药物纳米载体，需要在细胞质或特定的细胞器(如细胞核、溶酶体或线粒体)内发挥治疗作用。到目前为止，人们已经进行了多次尝试，但只有部分成功地将各种低分子和大分子药物和载药载体直接带入细胞质或单个细胞器，绕过内吞途径，保护药物免受溶酶体降解。由于各种药物纳米载体被越来越多地用于提高药物的有效性和安全性，这一应用的最终目标是设计一套能够将各种药物靶向输送到特定细胞内细胞器的纳米颗粒药物载体系统。我们推测，用能够靶向细胞内细胞器的特定配体修饰的载药纳米载体，如脂质体和胶束，将显著提高基因治疗、药物进入溶酶体和癌细胞凋亡的效率。为了验证这一假设，我们将追求以下具体目标：(1)制备具有不同配体的药物纳米载体(脂质体和聚乙二醇磷脂酰乙醇胺(PEGPE)共轭胶束)，以优先与选定的细胞内细胞器结合。然后，我们将研究它们与纯化的单个细胞器以及与正常和癌症小鼠和人类细胞的相互作用，以及它们在细胞内的分布和命运；(2)利用体外细胞培养系统，研究载药、模型化合物或DNA修饰的特定配体修饰的脂质体和胶束的生物学性质以及它们向细胞核、溶酶体和线粒体的输送；(3)使用体内肿瘤模型，研究药物或DNA修饰的特定配体修饰的纳米载体(脂质体和胶束)的生物学和治疗特性。我们期望这一应用能够成功地开发出一种新的平台，在细胞内将药物和基因定向输送到特定的细胞内细胞器，以提高治疗效果。 公共卫生相关性：这一应用的主要目标是解决细胞内细胞器特异性药物输送这一极其重要且仍然具有挑战性的问题，从而极大地提高包括抗癌治疗在内的许多疗法的疗效。我们的目标是设计一个平台，以创建一个能够在细胞内穿透并针对细胞内隔室和细胞器(如细胞核、溶酶体和线粒体)进行特定靶向的药物输送系统，以改善药物和DNA的输送。创建这种系统的算法应该适用于各种疾病和治疗方案。我们将通过使用新型和有前景的促凋亡抗癌药物的线粒体靶向实验癌症治疗，以及模型化合物和模型质粒的溶酶体和核的实验靶向来验证该方法的有效性。

DESCRIPTION (provided by applicant): Many pharmaceutical agents, including various large molecules (proteins, enzymes, antibodies) and even drug-loaded pharmaceutical nanocarriers, need to be delivered intracellularly to exert their therapeutic action within the cytoplasm or specific organelles, such as nuclei, lysosomes, or mitochondria. So far, multiple and only partially successful attempts have been made to brings various low-molecular-weight and macromolecular drugs and drug-loaded pharmaceutical carriers directly into the cell cytoplasm or individual organelles bypassing the endocytic pathway and protecting drugs from lysosomal degradation. Since, various pharmaceutical nanocarriers are increasingly used to increase the efficacy and safety of drugs, the final goal of this application is to engineer a set of nanoparticulate drug carrier systems capable of targeted delivery of various pharmaceuticals to specific intracellular organelles. We hypothesize that the application of drug-loaded pharmaceutical nanocarriers, such as liposomes and micelles, modified with the specific ligands capable of targeting intracellular organelles, such as nuclei, mitochondria, and lysosomes, will significantly increase the efficacy of gene therapy, drug delivery into lysosomes, and apoptosis in cancer cells. To check this hypothesis, we will pursue the following specific aims: (1) Prepare formulations of pharmaceutical nanocarriers (liposomes and polymeric polyethylene glycol-phosphatidylethanolamine (PEG- PE) conjugate-based micelles) with various ligands for preferential binding with selected intracellular organelles. We will then study their interaction with both purified individual cell organelles and with normal and cancer murine and human cells as well as their intracellular distribution and fate; (2) Investigate, using in vitro cell culture systems, biological properties of drug-, model compound-, or DNA-loaded specific ligand-modified liposomes and micelles and their cargo delivery into nuclei, lysosomes, and mitochondria; (3) Investigate, using in vivo tumor models, biological and therapeutical properties of drug-, or DNA-loaded specific ligand- modified nanocarriers (liposomes and micelles). We expect this application to successfully develop a novel platform for targeted drug and gene delivery within cells to specific intracellular organelles enhanced therapeutic outcome. PUBLIC HEALTH RELEVANCE: The main goal of this application is to solve the extremely important and still challenging problem of intracellular organelle-specific drug delivery, thus dramatically increasing the efficacy of many therapies, including anti-cancer therapies. We aim to engineer a platform for creating drug delivery systems capable of intracellular penetration and specific targeting of intracellular compartments and organelles, such as nuclei, lysosomes and mitochondria, for improved delivery of drugs and DNA. The algorithm for creating such systems should be applicable to a variety of diseases and treatment protocols. We will confirm the validity of the approach by testing with the mitochondria-targeted experimental cancer therapy using novel and promising proapoptotic anti-cancer drugs, and with experimental targeting of lysosomes and nuclei with model compounds and model plasmid.