ANTIVIRAL ACTIVITY OF A PHOSPHOROTHIOATE OLIGONUCLEOTIDE COMPLEMENTARY TO RNA OF THE HUMAN CYTOMEGALOVIRUS MAJOR IMMEDIATE-EARLY REGION
ANTIVIRAL ACTIVITY OF A PHOSPHOROTHIOATE OLIGONUCLEOTIDE COMPLEMENTARY TO RNA OF THE HUMAN CYTOMEGALOVIRUS MAJOR IMMEDIATE-EARLY REGION
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DOI：
10.1128/aac.37.9.1945
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发表时间：
1993-09-01
影响因子：
4.9
通讯作者：
ANDERSON, KP
中科院分区：
文献类型：
--
作者：
AZAD, RF;DRIVER, VB;ANDERSON, KP
在以原代人真皮成纤维细胞为宿主细胞的 96 孔免疫测定中，评估了与人巨细胞病毒 (HCMV) DNA 聚合酶基因的 mRNA 或 HCMV 主要立即早期区域 1 和 2（IE1 和 IE2）的 RNA 转录物互补的硫代磷酸寡核苷酸的抗病毒活性。与 IE2 区域 RNA 互补的寡核苷酸表现出最有效的抗病毒活性。其中一种寡核苷酸 ISIS 2922 的效力比核苷类似物更昔洛韦至少强 30 倍，在 96 孔免疫测定中的 50% 有效浓度为 0.37 μM。在感染性病毒产量减少测定中，ISIS 2922 和更昔洛韦在浓度为 2.2 和 36 muM 时分别将感染性病毒的产量减少了 2 个对数单位。对照寡核苷酸在浓度高达 3 μM 时未表现出对病毒产生的抑制作用。 ISIS 2922 以剂量依赖性方式减少 HCMV 感染细胞中的 IE 蛋白合成，这与抗病毒活性相关。 ISIS 2922 的抗病毒活性并非由寡核苷酸诱导的细胞毒性所致，因为仅在远远超过有效抗病毒浓度的浓度下才观察到对细胞活力或增殖的影响。 ISIS 2922 的特异性和效力表明它可能有助于治疗人类巨细胞病毒疾病。
Phosphorothioate oligonucleotides complementary to mRNA of the human cytomegalovirus (HCMV) DNA polymerase gene or to RNA transcripts of the major immediate-early regions 1 and 2 (IE1 and IE2) of HCMV were evaluated for antiviral activity in a 96-well immunoassay with primary human dermal fibroblasts as host cells. Oligonucleotides complementary to RNA of the IE2 region exhibited the most potent antiviral activity. One of these oligonucleotides, ISIS 2922, was at least 30-fold more potent than the nucleoside analog, ganciclovir, with a 50% effective concentration of 0.37 muM in the 96-well immunoassay. In an infectious virus yield reduction assay, ISIS 2922 and ganciclovir reduced production of infectious virus by 2 log units at concentrations of 2.2 and 36 muM, respectively. A control oligonucleotide showed no inhibition of virus production at concentrations as high as 3 muM. ISIS 2922 reduced IE protein synthesis in HCMV-infected cells in a dose-dependent manner which correlated with antiviral activity. The antiviral activity of ISIS 2922 was not due to oligonucleotide-induced cytotoxicity since effects on cell viability or proliferation were observed only at concentrations well in excess of effective antiviral concentrations. The specificity and potency of ISIS 2922 suggest that it may be useful for the treatment of cytomegalovirus disease in humans.