Nanoformulation Uptake, Release, Toxicology and Tissue Delivery

项目2摘要 该项目将长效抗逆转录病毒治疗性纳米制剂(NanoArt)合成、病毒组织 在啮齿动物和大型动物身上进行的水库靶向性和药效学试验 必填项。这项计划有三个共同目标。第一，寻求确定靶向药物的有效性 人源化HIV-1慢性感染小鼠持续抗逆转录病毒和免疫应答的配方 用装饰过的或有针对性的纳米艺术处理。病毒载量的减少和恢复的CD4+T细胞数量将是 与本地药物、含有非靶向ART的制剂和 用无关的对照药物(例如，氟康唑)治疗的动物。潜力的并行测试 组织和细胞毒性以及“推定的”药物-药物相互作用--药物滥用中的一个特殊问题 人口，将被确定。第二项研究旨在进一步评估最有效的药物颗粒 定量确定免疫细胞亚群在其组织部位(胃肠道相关)的目标 淋巴组织、骨髓、淋巴结和脑)含有残留病毒。第三个，将使用 抗逆转录病毒和免疫调节剂的组合(例如，吲哚胺2，3-双加氧酶 抑制物(DITO，GSK3176181A)诱导HIV-1gag/pol特异性细胞毒淋巴细胞，其目标可能是 根除病毒的能力。这些目标中每一个最有效的测试组合将在 SIV/SIV感染恒河猴。一种可注射的纳米艺术组合的开发 每6个月服用一次是现实的。Cabotegravir是一种已经长效的整合酶抑制剂，目前在 第二阶段试验。单次注射后的半衰期为21-50天，具有 有可能每三个月服用一次。我们在这个项目中提供的初步数据显示， 在药物储备量有更大增加的情况下，PK比正在给药的人有所改善 瞄准目标。这与拉米夫定和阿巴卡韦的聚合物包裹前体药物的产生相结合 显著延长的药物半衰期使联合长效治疗变得现实而不需要我们的掌握。在……上面 平衡，我们也承认存在障碍。但平衡了之前的科学成就，广泛的内部技术 经验和资源，包括训练有素的人员、设备和基础设施，可以应对挑战。 进一步的支持包括在葛兰素史克的支持下获得开发的和新的抗逆转录病毒药物 药品图书馆。纳米艺术将包括以单核巨噬细胞和亚细胞细胞器为靶向的细胞 (迟发和再循环的内体)。我们假设配方在尺寸、成分、涂层、 电荷将增强纳米艺术细胞的摄取，并增强随后的药物输送和释放到病毒中 水库。其中包括易受感染的CD4+T淋巴细胞。内布拉斯加州纳米医学中心的成立 生产工厂，一个现在位于医疗中心的良好的制造生产设施，使 改进了纳米艺术的放大生产。

Project 2 Abstract This project bridges long-acting antiretroviral therapeutic nanoformulation (nanoART) synthesis, viral tissue reservoir targeting and pharmacodynamic tests in rodents and large animal confirmatory studies when required. Three aims joint the project. The first, seeks to determine the effectiveness of the targeted drug formulations for sustained antiretroviral and immune responses in chronically HIV-1 infected humanized mice treated with decorated or targeted nanoART. Reductions in viral load and restored CD4+ T cell numbers will be assessed when compared against results seen with native drug, formulations containing untargeted ART and those animals treated with an irrelevant control drug (for example, fluconazole). Parallel tests for potential tissue and cellular toxicities and “putative” drug-drug interactions, a special concern in the drug-abusing population, will be determined. The second seeks to evaluate further the most effective drug particles with the goal of quantitatively determining the subset of immunocytes in its tissue locale (gastrointestinal associated lymphoid tissue, bone marrow, lymph node and brain) that harbor residual virus. The, third will use combinations of antiretroviral and immune modulatory agents (for example, an indoleamine 2,3-dioxygenase inhibitor (IDIO, GSK3176181A) to induce HIV-1gag/pol-specific cytotoxic lymphocytes with the “putative” goal of viral eradication. The most effective tested combinations for each of these aims will be cross-validated in SIV/SHIV infected rhesus macaques. The development of an injectable combination nanoART that can be dosed once every 6 months is realistic. Cabotegravir, is an already long-acting integrase inhibitor, currently in phase II trials. With a demonstrated half-life of 21–50 days following a single parenteral dose, it has the potential for an every three-month dose. Our own preliminary data offered in this project shows an up to 5-fold improvement in PK from what is being administered to eople with an even larger increase in drug reservoir targeting. This combined with the generation of polymer-encased pro-drugs of lamivudine and abacavir with markedly extended drug half-lives makes combination long-acting therapy realistic and without our grasp. On balance, we also acknowledge the obstacles. But balanced prior scientific successes, broad in-house technical experience and resources including trained personnel, equipment and infrastructure can meet the challenges. Further support includes access to developed and new antiretrovirals with support from GlaxoSmithKline and drug libraries. NanoART will include cell targeting to monocyte-macrophages and to subcellular organelles (late and recycling endosomes). We posit that modifications made of formulations in size, composition, coating, and charge will enhance nanoART cell uptake and potentiate subsequent drug delivery and release into viral reservoirs. These include susceptible CD4+ T lymphocytes. The establishment of the Nebraska Nanomedicine Production Plant, a good manufacturing production facility now housed at the medical center has enabled improved scale-up production of nanoART.