Vinogradov Serguei V, Poluektova Larisa Y, Makarov Edward, Gerson Trevor, Senanayake Madapathage T
Department of Pharmaceutical Sciences and Center for Drug Delivery and Nanomedicine, University of Nebraska Medical Center, Omaha, NE, USA.
Antivir Chem Chemother. 2010 Oct 28;21(1):1-14. doi: 10.3851/IMP1680.
Macrophages serve as a depot for HIV type-1 (HIV-1) in the central nervous system. To efficiently target macrophages, we developed nanocarriers for potential brain delivery of activated nucleoside reverse transcriptase inhibitors (NRTIs) called nano-NRTIs.
Nanogel carriers consisting of poly(ethylene glycol) (PEG)- or Pluronic-polyethylenimine (PEI) biodegradable networks, star PEG-PEI or poly(amidoamine) dendrimer-PEI-PEG dendritic networks, as well as nanogels decorated with brain-targeting peptide molecules, specifically binding to the apolipoprotein E receptor, were synthesized and evaluated. Nano-NRTIs were obtained by mixing aqueous solutions of zidovudine 5'-triphosphate or didanosine 5'-triphosphate and nanocarriers, followed by freeze-drying. Intracellular accumulation, cytotoxicity and antiviral activity of nano-NRTIs were monitored in monocyte-derived macrophages (MDMs). HIV-1 viral activity in infected MDMs was measured by a reverse transcriptase activity assay following treatment with nano-NRTIs. Mitochondrial DNA depletion in MDMs and human HepG2 cells was assessed by quantitative PCR.
Nanogels were efficiently captured by MDMs and demonstrated low cytotoxicity, and no antiviral activity without drugs. All nano-NRTIs demonstrated high efficacy of HIV-1 inhibition at drug levels as low as 1 μmol/l, representing a 4.9- to 14-fold decrease in 90% effective drug concentrations as compared with NRTIs, whereas 50% cytotoxicity effects started at 200× higher concentrations. Nano-NRTIs with a core-shell structure and decorated with brain-targeting peptides displayed the highest antiviral efficacy. Mitochondrial DNA depletion, a major cause of NRTI neurotoxicity, was reduced threefold compared with NRTIs at application of selected nano-NRTIs.
Nano-NRTIs demonstrated a promising antiviral efficacy against HIV-1 in MDMs and showed strong potential as nanocarriers for delivery of antiviral drugs to macrophages harbouring in the brain.
巨噬细胞是1型人类免疫缺陷病毒(HIV-1)在中枢神经系统中的储存库。为了有效靶向巨噬细胞,我们开发了用于潜在脑内递送活化核苷类逆转录酶抑制剂(NRTIs)的纳米载体,称为纳米-NRTIs。
合成并评估了由聚乙二醇(PEG)或普朗尼克-聚乙烯亚胺(PEI)可生物降解网络、星状PEG-PEI或聚(酰胺胺)树枝状聚合物-PEI-PEG树枝状网络组成的纳米凝胶载体,以及用与载脂蛋白E受体特异性结合的脑靶向肽分子修饰的纳米凝胶。纳米-NRTIs通过将齐多夫定5'-三磷酸或去羟肌苷5'-三磷酸的水溶液与纳米载体混合,然后冻干获得。在单核细胞衍生的巨噬细胞(MDMs)中监测纳米-NRTIs的细胞内积累、细胞毒性和抗病毒活性。在用纳米-NRTIs处理后,通过逆转录酶活性测定法测量感染MDMs中的HIV-1病毒活性。通过定量PCR评估MDMs和人HepG2细胞中的线粒体DNA耗竭情况。
纳米凝胶被MDMs有效摄取,显示出低细胞毒性,且在无药物时无抗病毒活性。所有纳米-NRTIs在低至1μmol/l的药物水平下均表现出对HIV-1的高效抑制作用,与NRTIs相比,90%有效药物浓度降低了4.9至14倍,而50%细胞毒性作用在高200倍的浓度时开始出现。具有核壳结构并修饰有脑靶向肽的纳米-NRTIs显示出最高的抗病毒功效。在应用选定的纳米-NRTIs时,线粒体DNA耗竭(NRTI神经毒性的主要原因)与NRTIs相比降低了三倍。
纳米-NRTIs在MDMs中对HIV-1显示出有前景的抗病毒功效,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。