Surface-modified pharmaceutical nanocarriers for subcellular targeting

描述（由申请人提供）：许多药剂，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，已经进行了多次且仅部分成功的尝试，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，并保护药物免受溶酶体降解。由于各种药物纳米载体越来越多地用于提高药物的功效和安全性，因此本申请的最终目标是设计一组能够将各种药物靶向递送至特定细胞内细胞器的纳米颗粒药物载体系统。我们假设，应用载药的药物纳米载体，如脂质体和胶束，修饰有能够靶向细胞内细胞器，如细胞核，线粒体和溶酶体的特异性配体，将显着增加基因治疗，药物递送到溶酶体，和癌细胞凋亡的疗效。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（脂质体和基于聚合物聚乙二醇-磷脂酰乙醇胺（PEG-PE）缀合物的胶束）的制剂，用于优先结合所选择的细胞内细胞器。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细胞的相互作用，以及它们的细胞内分布和命运;（2）使用体外细胞培养系统，研究负载药物、模型化合物或DNA的特异性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、溶酶体和线粒体的货物递送;（3）使用体内肿瘤模型研究药物或DNA负载的特异性配体修饰的纳米载体（脂质体和胶束）的生物学和治疗性质。我们希望这一应用能够成功地开发出一种新的平台，用于靶向药物和基因在细胞内递送到特定的细胞内细胞器，从而增强治疗效果。 公共卫生相关性：该应用的主要目标是解决细胞内细胞器特异性药物递送的极其重要且仍然具有挑战性的问题，从而显著提高许多疗法（包括抗癌疗法）的功效。我们的目标是设计一个平台，用于创建能够细胞内渗透和特异性靶向细胞内区室和细胞器（如细胞核，溶酶体和线粒体）的药物递送系统，以改善药物和DNA的递送。创建这种系统的算法应该适用于各种疾病和治疗方案。我们将通过使用新的和有前途的促凋亡抗癌药物的靶向实验性癌症治疗，以及使用模型化合物和模型质粒的溶酶体和细胞核的实验靶向测试来确认该方法的有效性。

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.