Pharmacology of Antiretroviral Nanoparticle Micelles

描述(由申请人提供)：抗逆转录病毒药物(ART)的使用，即高效抗逆转录病毒疗法(HAART)的使用，大大降低了与人类免疫缺陷病毒1型(HIV-1)疾病相关的死亡率。然而，治疗失败、给药的复杂性和不良反应都限制了感染者的长期治疗。单核巨噬细胞不仅是HIV-1的早期宿主，也是传播HIV-1的载体。基于以前对吲哚那韦纳米粒(NP)的研究，我们希望将我们的工作扩展到包括联合抗逆转录病毒药物，并利用药物的组合来制造纳米粒。纳米制剂可以支持将药物持续释放到被视为避难所的HIV-1组织，并提供远远超过抑制商的药物浓度。我们假设，组合ARV纳米配方药物传递系统可以优化，以从身体的受限区域根除HIV-1。使用可生物降解的聚合物(聚(DL-丙交酯-5-己内酯))，复合抗逆转录病毒制剂(利托那韦RTV、洛匹那韦LPV和EFV)将被包埋到纳米颗粒中，并将考察其效率。课程内容包括：高效液相色谱、扫描电子显微镜和共聚焦显微镜，以及巨噬细胞作为药物输送系统的功能研究。体外中空纤维模型将通过p24抗原检测和高效液相色谱检测ART NP药物水平来探讨抑制HIV-1的药效学。对巨噬细胞的渗透性将用液相色谱、共聚焦和透射电子显微镜进行评估。这些纳米制剂将被用作药物输送系统，以提高ART对小鼠网状内皮系统(RES；脾、肝、淋巴结)的组织渗透率。复合抗逆转录病毒纳米粒的药效学和药代动力学将在更长的时间内使用体外和体内系统进行测定，与游离药物和空白纳米粒相比。这项提议的目的是更好地了解如何优化使用NP药物输送技术，以抑制艾滋病毒-1藏身的人体受限区域产生病毒。这些方法为艾滋病毒-1的治疗提供了直接的患者利益。 公共卫生相关性：这项应用将研究在细胞培养系统和动物中被制成纳米粒的联合抗逆转录病毒药物(利托那韦、洛匹那韦和依法韦仑)的药理学。这些纳米颗粒将被运送到HIV-1避难所(脾、肝、淋巴结)。由于纳米颗粒尺寸较小，颗粒在组织中停留的时间更长，并允许较少的频率给药。这种纳米颗粒传递系统可能会影响HIV-1治疗的未来。

DESCRIPTION (provided by applicant): The use of anti-retroviral agents (ART), namely highly active anti-retroviral therapy (HAART) has significantly diminished the mortality associated with human immunodeficiency virus type-1 (HIV-1) disease. However, treatment failures, dosing complexities, and adverse effects all limit the long-term therapy in infected peoples. Mononuclear phagocytes serve as not only as early reservoirs but also vehicles for dissemination for HIV-1. Based on previous work with indinavir nanoparticles (NP) we want to extend our work to include combination ARV agents and fabricate NP with combinations of drugs. Nanoformulations can support sustained drug release to HIV-1 tissues considered sanctuaries and deliver drug concentrations that far exceed the inhibitory quotient. We hypothesize that combination ARV nanoformulated drug delivery system can be optimized to eradicate HIV-1 from restricted areas of the body. Using a biodegradable polymer (poly (DL-lactide-co 5-caprolactone)), combined ARV agents (ritonavir RTV, lopinavir LPV, and efavirenz EFV) will be entrapped into a nanoparticle and efficiency will be investigated. Liquid chromatography, scanning electron and confocal microscopy, and functional studies in macrophages as a drug delivery system will be gathered. A hollow fiber in vitro model will explore the pharmacodynamics of HIV- 1 inhibition as measured by p24 antigen assay and ART NP drug levels measured by liquid chromatography. Penetration into macrophages will be assessed by liquid chromatography and confocal and transmission electron microscopy. The nanoformulations will be used as a drug delivery system to improve tissue penetrance of ART into the reticuloendothelial system (RES; spleen, liver, lymph nodes) in mice. The pharmacodynamics and pharmacokinetics of combination ARV NP will be determined using in vitro and in vivo systems over extended time periods as compared to free soluble drugs as well as blank nanoparticles. The aims of this proposal are a better understanding of how to optimize the use of NP drug delivery technology to inhibit the production of virus from restricted areas of the body where HIV-1 hides. These approaches offer direct patient benefit for the treatment of HIV-1. PUBLIC HELATH RELEVANCE: This application will investigate the pharmacology of combination antiretrovirals (ritonavir, lopinavir, and efavirenz) fabricated into nanoparticles both in cell culture systems and in animals. The nanoparticles will be delivered to HIV-1 sanctuaries (spleen, liver, lymph nodes). Due to the nanoparticle small size, the particles will stay in tissue longer and allow for less frequent dosing. This nanoparticle delivery system could affect the future of HIV- 1 treatment.