Surface-modified pharmaceutical nanocarriers for subcellular targeting

描述（由申请人提供）：许多药物制剂，包括各种大分子（蛋白质、酶、抗体），甚至是载药的药物纳米载体，需要在细胞内递送，以在细胞质或特定细胞器（如细胞核、溶酶体或线粒体）内发挥其治疗作用。到目前为止，将各种低分子量和大分子药物以及载药药物载体绕过内吞途径直接进入细胞质或单个细胞器，保护药物不被溶酶体降解，已经进行了多次且仅部分成功的尝试。由于各种药物纳米载体越来越多地用于提高药物的有效性和安全性，因此该应用的最终目标是设计一套纳米颗粒药物载体系统，能够将各种药物靶向递送到特定的细胞内细胞器。我们假设，应用载药的药物纳米载体，如脂质体和胶束，通过能够靶向细胞内细胞器（如细胞核、线粒体和溶酶体）的特定配体修饰，将显著提高基因治疗的疗效，药物递送到溶酶体，以及癌细胞的凋亡。为了验证这一假设，我们将追求以下具体目标：(1)制备具有各种配体的药物纳米载体（脂质体和聚合物聚乙二醇-磷脂酰乙醇胺（PEG- PE）缀合物胶束）的配方，以优先结合选定的细胞内细胞器。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，研究药物、模型化合物或dna负载的特异性配体修饰脂质体和胶束的生物学特性，以及它们向细胞核、溶酶体和线粒体的运送；(3)利用体内肿瘤模型，研究药物或dna负载的特异性配体修饰纳米载体（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一个新的平台，将靶向药物和基因在细胞内传递到特定的细胞内细胞器，从而提高治疗效果。

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.