Antiviral Efficacy of the Novel Compound BIT225 against HIV-1 Release from Human Macrophages
Antiviral Efficacy of the Novel Compound BIT225 against HIV-1 Release from Human Macrophages
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DOI：
10.1128/aac.01308-09
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发表时间：
2010-02-01
影响因子：
4.9
通讯作者：
Wilkinson, John
中科院分区：
文献类型：
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作者：
Khoury, Gabriela;Ewart, Gary;Wilkinson, John
基于先前发现的阿米洛利类似物抑制单核细胞源性巨噬细胞（MDM）中HIV-1的复制，百特龙有限公司（Biotron Limited）建立了一个包含300多种小分子化合物的文库，这些化合物在抗HIV-1活性方面有显著提高。我们的先导化合物BIT225阻断Vpu离子通道活性，也显示出抗hiv -1活性，50%有效浓度为2.25 +/- 0.23 μ M（平均+/-标准误差），在感染MDM中的体外毒性最小（50%毒性浓度，284 μ M），选择性指数为126。在这项研究中，我们确定了BIT225活性在巨噬细胞中的抗逆转录病毒疗效，巨噬细胞是重要的药物靶点，因为单核细胞系的细胞是HIV-1的关键储存库，在它们分化成巨噬细胞时将病毒传播到外周组织。在急性和慢性HIV-1（Ba-L）感染MDM的实验中，通过实时PCR和逆转录酶（RT）活性测定，BIT225在单核细胞向巨噬细胞分化的各个阶段显著降低了病毒整合和病毒释放。此外，TZM-bl试验表明，在存在BIT225的情况下产生的低水平新生病毒的传染性低于不存在该化合物的情况下产生的病毒。在缺乏Vpu的慢性感染HIV-2的MDM中没有观察到抗病毒活性，证实了我们最初针对该病毒蛋白的靶向和筛选。BIT225的活性是病毒后整合，对HIV-1酶RT和蛋白酶没有直接影响。本研究结果表明，BIT225是一种针对Vpu的病毒生命周期晚期抑制剂，是一种能够显著抑制急性和慢性感染巨噬细胞中HIV-1释放的药物。
Building on previous findings that amiloride analogues inhibit HIV-1 replication in monocyte-derived macrophages (MDM), Biotron Limited has generated a library of over 300 small-molecule compounds with significant improvements in anti-HIV-1 activity. Our lead compound, BIT225, blocks Vpu ion channel activity and also shows anti-HIV-1 activity, with a 50% effective concentration of 2.25 +/- 0.23 mu M (mean +/- the standard error) and minimal in vitro toxicity (50% toxic concentration, 284 mu M) in infected MDM, resulting in a selectivity index of 126. In this study, we define the antiretroviral efficacy of BIT225 activity in macrophages, which are important drug targets because cells of the monocyte lineage are key reservoirs of HIV-1, disseminating virus to the peripheral tissues as they differentiate into macrophages. In assays with acutely and chronically HIV-1(Ba-L)-infected MDM, BIT225 resulted in significant reductions in viral integration and virus release as measured by real-time PCR and a reverse transcriptase (RT) activity assay at various stages of monocyte-to-macrophage differentiation. Further, the TZM-bl assay showed that the de novo virus produced at low levels in the presence of BIT225 was less infectious than virus produced in the absence of the compound. No antiviral activity was observed in MDM chronically infected with HIV-2, which lacks Vpu, confirming our initial targeting of and screening against this viral protein. The activity of BIT225 is post-virus integration, with no direct effects on the HIV-1 enzymes RT and protease. The findings of this study suggest that BIT225 is a late-phase inhibitor of the viral life cycle, targeting Vpu, and is a drug capable of significantly inhibiting HIV-1 release from both acute and chronically infected macrophages.