SYNTHESIS & ASSEMBLY OF BIO-RESPONSIVE COPOLYMER VESICLES FOR PAYLOAD TRANSPORT

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 对治疗效果至关重要。不幸的是，许多药物输送方法往往是高度 由于血清稳定性不理想，转运跨越生物屏障，并在目标部位释放，效率低下。 为了解决这些问题，我们将创造生物响应性的，聚合物纳米容器为选择性 将治疗性化合物输送到预先确定的细胞位置。我们将结合解决方案自组装 具有选择性靶向和裂解多肽连接以允许囊泡的嵌段共聚物 对关键环境刺激的响应进化，最终导致特定部位释放 封装的有效载荷。 在第一个具体目标中，我们将创造新型的含肽嵌段共聚物和 表征它们在水溶液中的自组装。这些两亲性共聚物将被设计成 组装成泡状结构。旨在促进内吞摄取的多肽(1)和内吞 释放物(2)将顺序地结合到嵌段共聚物的亲水主链中 层次化的时尚。在第二个具体目标中，我们将评估蛋白酶的敏感性和靶向性。 各多肽层的效率。蛋白水解酶的可及性，每个肽序列，以及小泡 稳定性，将被评估为囊泡日冕中肽位置的函数。细胞-囊泡结合， 囊泡内化和囊泡的内溶活性也将被评估。在第三节 特定的目的，我们将验证多肽引导囊泡运输和破裂的能力。 在胞质内，我们将评估无有效载荷和细胞毒素掺入的细胞毒性 水泡。通过选择性地将多肽(2)放置在近端将导致囊泡破裂。 聚合物的疏水嵌段。 我们对这个项目的长期愿景是开发一种核酸输送系统，用于 选择性靶向肿瘤间质成纤维细胞和/或炎症细胞。我们设想了一个完整的 NOVO设计的模块化囊泡纳米胶囊，包含一系列位置特定的“可脱落”壳 在每个运输障碍处响应环境提示来指挥有效载荷运输。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. critically important for therapeutic efficacy. Unfortunately, many methods for drug delivery are often highly inefficient due to non-ideal serum-stability, transport across biological barriers, and release at the target site. To address these problems, we will create bio-responsive, polymeric nano-containers for the selective delivery of therapeutic compounds to pre-determined cellular locations. We will combine the solution selfassembly of block copolymers with selective targeting and cleaving peptide linkages to permit vesicle evolution in response to critical environmental stimuli, eventually leading to site-specific release of the encapsulated payload. In the first specific aim, we will create novel peptide-containing block copolymers and characterize their self-assembly in aqueous solution. These amphiphilic copolymers will be designed to assemble into vesicular structures. Peptides designed to promote endocytotic uptake (1) and endosomal release (2) will be sequentially incorporated into the hydrophilic backbone of the block copolymer in a layered fashion. In the second specific aim, we will evaluate the protease-sensitivity and targeting efficiency of each peptide layer. The protease accessibility of each peptide sequence, as well as vesicle stability, will be assessed as a function of peptide position in the vesicle's corona. The cell-vesicle binding, vesicle internalization, and endosomolytic activity of the vesicles also will be evaluated. In the third specific aim, we will validate the ability of the peptides to direct vesicle transport to and rupture within the cytosol, and we will evaluate the cytotoxicity of both payload-free and cytotoxinincorporating vesicles. Vesicle rupture will be induced by selective placement of peptide (2) proximal to the hydrophobic block of the polymer. Our long-term vision for this project is the development of a nucleic acid delivery system for the selective targeting of tumor stromal fibroblasts and/or inflammatory cells. We envision the complete de novo design of modular vesicular nano-capsules containing a series of location specific "sheddable" shells to direct payload transport in response to environmental cues at each transport barrier.