Nano-NRTIs: efficient inhibitors of HIV type-1 in macrophages with a reduced mitochondrial toxicity.
Nano-NRTIs: efficient inhibitors of HIV type-1 in macrophages with a reduced mitochondrial toxicity.
复制标题
DOI：
10.3851/imp1680
复制
发表时间：
2010-10-28
影响因子：
--
通讯作者：
Senanayake MT
中科院分区：
文献类型：
--
作者：
Vinogradov SV;Poluektova LY;Makarov E;Gerson T;Senanayake MT
巨噬细胞作为HIV-1在中枢神经系统（CNS）中的储存库。为了有效地靶向巨噬细胞，我们开发了用于活化核苷逆转录酶抑制剂（Nano-NRTI）的潜在脑递送的纳米载体。合成并评价了由PEG-或Pluronic-PEI可生物降解网络、星星PEG-PEI或PAMAM-PEI-PEG树枝状网络组成的纳米凝胶载体，以及用多个ApoE肽分子修饰的纳米凝胶，其特异性结合载脂蛋白E受体。通过混合三磷酸盐（AZTTP或ddITP）和纳米载体的水溶液，然后冷冻干燥来获得纳米NRTI。在单核细胞衍生的巨噬细胞（MDM）中监测Nano-NRTI的细胞内蓄积、细胞毒性和抗病毒活性。在用Nano-NRTI处理后，通过微量RT测定法测量感染的MDM中的HIV-1ADA病毒活性。通过定量PCR（qPCR）评估MDM和人HepG 2细胞中的线粒体DNA（mtDNA）耗竭。纳米凝胶被MDM有效地捕获，并表现出低细胞毒性，在没有药物的情况下不影响病毒活性。所有Nano-NRTI在低至1 μmol/L的药物水平下均表现出高的HIV-1抑制效力，与NRTI相比，有效药物浓度（EC 90）降低了4.9至14倍，而细胞毒性效应（IC 50）开始于高200倍的浓度。具有核-壳结构并用载体肽（例如脑靶向ApoE肽）修饰的纳米载体显示出最高的抗病毒功效。在应用选定的Nano-NRTI时，与NRTI相比，mtDNA耗竭（NRTI神经毒性的主要原因）降低了3倍。纳米NRTI在MDM中表现出有希望的抗病毒功效，并显示出作为纳米载体将抗病毒药物递送至脑内巨噬细胞的强大潜力。
Macrophages serve as depot for HIV-1 in the central nervous system (CNS). To efficiently target macrophages, we developed nanocarriers for potential brain delivery of activated nucleoside reverse transcriptase inhibitors (Nano-NRTI). Nanogel carriers consisting of PEG- or Pluronic-PEI biodegradable networks, star PEG-PEI, or PAMAM-PEI-PEG dendritic networks, as well as nanogels decorated with multiple ApoE peptide molecules, specifically binding to the apolipoprotein E receptor, were synthesized and evaluated. Nano-NRTIs were obtained by mixing aqueous solutions of triphosphates (AZTTP or ddITP) and nanocarriers followed by freeze-drying. Intracellular accumulation, cytotoxicity, and antiviral activity of Nano-NRTIs were monitored in monocyte-derived macrophages (MDMs). HIV-1ADA viral activity in infected MDMs was measured by micro-RT assay following the treatment with Nano-NRTIs. Mitochondrial DNA (mtDNA) depletion in MDMs and human HepG2 cells was assessed by quantitative PCR (qPCR). Nanogels were efficiently captured by MDMs and demonstrated low cytotoxicity, not affecting viral activity without drugs. All Nano-NRTIs demonstrated high efficacy of HIV-1 inhibition at drug levels as low as 1 μmol/L, representing from 4.9 to 14-fold decrease in effective drug concentrations (EC90) as compared to NRTIs, while cytotoxicity effects (IC50) started at 200 times higher concentrations. Nanocarriers with core-shell structure and decorated with vector peptides (e.g. brain-targeting ApoE peptide) displayed the highest antiviral efficacy. The mtDNA depletion, a major cause of NRTI neurotoxicity, was reduced 3-fold compared to NRTIs at application of selected Nano-NRTIs. Nano-NRTIs demonstrated a promising antiviral efficacy in MDMs and showed strong potential as nanocarriers for delivery of antiviral drugs to brain-harboring macrophages.