Nanomedicine and NeuroAIDS

描述(由申请人提供)：在其中枢神经系统(CNS)避难所内消除人类免疫缺陷病毒受到以下因素的影响：抗逆转录病毒疗法(ART)对血脑屏障(BBB)的可变渗透率、复杂的给药方案、成本、毒性以及生物分布和药代动力学药物模式的限制。尽管抗逆转录病毒疗法及其导致脑脊液病毒载量显著减少的能力取得了进展，但神经艾滋病的发病率仍在继续上升。实现最大临床反应的一个主要问题障碍是在受疾病影响的脑区维持较高的ART药物水平。为了解决这个问题，我们将开发常用抗逆转录病毒药物(洛比那韦、利托那韦和依法韦仑)的纳米制剂，并在循环中通过血液传播的单核细胞-巨噬细胞递送药物。改善BBB内ART传播的方法将需要分三步走。首先，将测试纳米粒(NP)药物制剂进入和分泌骨髓来源的巨噬细胞(BMM)和单核细胞来源的巨噬细胞(MDM)的比较措施。在这里，病毒蛋白酶和非核苷类逆转录酶抑制剂(S)将被包装成磷脂包裹的NP。将测量载药颗粒的细胞毒性、抗逆转录病毒疗效、流动性和巨噬细胞携带的功能后果。其次，这些制剂的药代动力学(摄取、释放、血浆和组织分布)将以BMM作为药物输送系统在小鼠体内进行研究。第三，将开发配体形式的NP并在体外进行测试，然后用于在小鼠体内直接静脉给药。或者，为了促进NP进入巨噬细胞，将用叶酸涂层制成配方，该涂层将专门针对巨噬细胞。将开展反映人类单核细胞和MDM免疫激活的实验室实验，以评估增强配体包衣NP制剂摄取的最佳方法。通过这种方式，药物绕过网状内皮系统并穿越血脑屏障的能力将被确定。高效液相色谱分析将被用来测量接受NP治疗的小鼠的脾、淋巴结、肝脏和大脑中的药物水平，并将提供药物组织渗透性的确认。这些测试将与组织学和成像分析一起使用。最后，开发的NP将在受影响的大脑中测试人类HIV-1中枢神经系统疾病的原始和人源化小鼠模型的抗逆转录病毒效果。总而言之，我们的目标是提高ART的治疗效果和血脑屏障迁移率，以便将它们转化为人类使用，以改善神经艾滋病的疾病结局。公共卫生意义尚未在其中枢神经系统庇护所内消灭人类免疫缺陷病毒，艾滋病毒-1痴呆症仍然是一个公共卫生问题。原因围绕着抗逆转录病毒疗法(ART)对血脑屏障的不同渗透率以及复杂的给药方案、成本和药物毒性。为了解决这个问题，我们计划开发具有不同脑进入曲线的常用ART的纳米制剂，并将它们直接输送到血源性巨噬细胞内的患病脑组织。

DESCRIPTION (provided by applicant): The elimination of the human immunodeficiency virus inside its central nervous system (CNS) sanctuary is affected by variable antiretroviral therapy (ART) penetrance across the blood-brain barrier (BBB), complex dosing regimens, costs, toxicities, and limitations in biodistribution and pharmacokinetic drug patterns. Despite advances in ART and its abilities to cause significant reductions in cerebrospinal fluid viral loads; NeuroAIDS morbidities continue on the rise. A principal issue obstacle in achieving maximal clinical responses is in maintaining high ART drug levels in disease affected brain subregions. To address this issue, we will develop nanformulations of commonly used anti-retroviral drugs (lopinavir, ritonavir, and efavirenz) and deliver the drugs inside circulating blood-borne monocyte-macrophages. The means to improve distribution of ART across the BBB will require a three-step approach. First, comparative measures of nanoparticle (NP) drug formulations will be tested for entry and secretion into and from bone marrow-derived macrophages (BMM) and monocyte-derived macrophages (MDM). Here, viral protease and nonnucleoside reverse transcriptase inhibitor(s) will be packaged into phospholipids coated NP. Cytotoxicity, anti-retroviral efficacy, mobility, and the functional consequences of macrophage carriage of the drug-laden particles will be measured. Second, pharmacokinetics (uptake, release, plasma and tissue distribution) of the formulations will be investigated using BMM as a drug delivery system in mice. Third, ligand-formulated NP will be developed and tested in vitro then used to test direct intravenous administration in mice. Alternatively and to enhance NP entry into macrophages, formulations will be made with folate coatings will be designed to specifically target macrophages. Laboratory experiments reflecting immune activation of human monocytes and MDM will be developed to assess the optimal ways to enhance uptake of ligand-coated NP formulations. In this way, the abilities of drug to bypass the reticuloendothelial system and cross the BBB will be determined. High performance liquid chromatography analyses will be used to measure drug levels in spleen, lymph nodes, liver and brain in NP-treated mice and will provide confirmation of drug tissue penetrance. These tests will be used in tandem with histology and imaging assays. Lastly, the NP developed will be tested for anti-retroviral efficacy in affected brains of a primary and humanized mouse models of human HIV-1 CNS disease. All together, the goals are to enhance therapeutic efficacy and BBB migration of ART so that they can be translated for human use to improve disease outcomes in NeuroAIDS. PUBLIC HEALTH RELEVANCE The elimination of the human immunodeficiency virus inside its central nervous system sanctuary has not been achieved and HIV-1 dementia remains a public health problem. The reasons revolve around variable antiretroviral therapy (ART) penetrance across the blood-brain barrier as well as complex dosing regimens, costs, and drug toxicities. To address this issue, we plan to develop nanoformulations of commonly used ART with variable brain entry profiles and deliver them directly to diseased brain tissue inside blood-borne macrophages.