Transplantation Virology, Institute for Medical Microbiology, Department of Biomedicine, University of Basel, and Infectious Diseases & Hospital Epidemiology, University Hospital Basel, Petersplatz 10, CH-4003 Basel, Switzerland.
Antimicrob Agents Chemother. 2011 May;55(5):2129-36. doi: 10.1128/AAC.00046-11. Epub 2011 Mar 14.
Polyomavirus JC (JCV) replication causes progressive multifocal leukoencephalopathy (PML), a frequently fatal brain disease in immunodeficient patients, yet antiviral drugs are lacking. We characterized the lipid conjugate 1-O-hexadecyloxypropyl-cidofovir (CMX001) regarding JCV (Mad-4) replication in human brain progenitor-derived astrocytes (PDA) and the simian virus 40 (SV40) large-T-antigen-expressing COS-7 cells up to 7 days postinfection (dpi). We examined JCV loads by PCR, the infection rate by immunofluorescence, and host cell toxicity by WST-1 and BrdU incorporation assays. Supernatants from CMX001-treated PDA demonstrated a drug concentration-dependent decrease in JCV loads and infectivity. CMX001 had only a modest effect on host cell metabolism but reduced overall BrdU incorporation. In PDA at 7 dpi, the CMX001 50% effective concentration (EC50) was 5.55 nM, the 50% cytotoxic concentration (CC50) was 184.6 nM, and the 50% selectivity index (SI50) was 33.3. The EC90 was 19.7 nM, the CC90 was 5,054 nM, and the SI90 was 256.1. In COS-7 cells, JCV replication was faster and the EC50 and EC90 were 18- and 37-fold higher than those in PDA, i.e., 0.1 μM and 0.74 μM (CC50, 0.67 μM; SI50, 6.7; CC90, 12.2 μM; SI90, 16.5) at 5 dpi. We conclude that CMX001 inhibits JCV replication at concentrations in vitro that can be attained by oral administration without significant side effects in clinical studies.
多瘤病毒 JC（JCV）复制会导致进行性多灶性白质脑病（PML），这是一种免疫缺陷患者中经常致命的脑部疾病，但目前缺乏抗病毒药物。我们对脂质缀合物 1-O-十六烷基氧基丙基-西多福韦（CMX001）进行了表征，研究了其在人脑祖细胞衍生的星形胶质细胞（PDA）和表达猿猴病毒 40（SV40）大 T 抗原的 COS-7 细胞中对 JCV（Mad-4）复制的作用，感染后时间长达 7 天（dpi）。我们通过 PCR 检测 JCV 负荷，通过免疫荧光检测感染率，通过 WST-1 和 BrdU 掺入测定法检测宿主细胞毒性。CMX001 处理的 PDA 上清液显示出药物浓度依赖性的 JCV 负荷和感染性降低。CMX001 对宿主细胞代谢仅有适度影响，但总体上降低了 BrdU 掺入。在 7dpi 的 PDA 中，CMX001 的 50%有效浓度（EC50）为 5.55 nM，50%细胞毒性浓度（CC50）为 184.6 nM，50%选择性指数（SI50）为 33.3。EC90 为 19.7 nM，CC90 为 5054 nM，SI90 为 256.1。在 COS-7 细胞中，JCV 复制速度更快，EC50 和 EC90 分别比 PDA 高 18 倍和 37 倍，即 0.1μM 和 0.74μM（CC50，0.67μM；SI50，6.7；CC90，12.2μM；SI90，16.5），在 5dpi。我们得出结论，CMX001 以体外可达到的浓度抑制 JCV 复制，而在临床研究中口服给药不会产生明显的副作用。
