A novel nonnucleoside inhibitor specifically targets cytomegalovirus DNA maturation via the UL89 and UL56 gene products
A novel nonnucleoside inhibitor specifically targets cytomegalovirus DNA maturation via the UL89 and UL56 gene products
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DOI：
10.1128/jvi.75.19.9077-9086.2001
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发表时间：
2001-10-01
影响因子：
5.4
通讯作者：
Hallenberger, S
中科院分区：
文献类型：
--
作者：
Buerger, I;Reefschlaeger, J;Hallenberger, S
3-Hydroxy-2,2-二甲基-N-[4({[5-(二甲氨基)-1-萘基]磺酰基}氨基)-苯基]丙酰胺 (BAY 38-4766) 是一种新型选择性非核苷巨细胞病毒 (CMV) 复制抑制剂，具有出色的安全性。该化合物和结构类似物既不抑制病毒 DNA 合成，也不抑制病毒转录和翻译。致密体和非感染性包膜颗粒的积累与基因组间转变时串联体加工和功能裂解的抑制同时发生，表明对病毒 DNA 成熟和单体基因组长度包装的干扰。在体外选择了耐药病毒群体，包括药物敏感性降低 566 倍的鼠巨细胞病毒 (MCMV) 分离株。对已知主要参与病毒 DNA 切割和包装的 6 个开放阅读框 (ORF) 进行测序，确定了 ORF UL56、UL89 和 UL104 中的突变。表达突变体 UL56、UL89 和 UL104 的鼠同源物的不同组合的 MCMV 重组体的构建以及药物敏感性分析清楚地表明，突变体 ORF UL89 外显子 II (M360I) 和 M56 (P202A I208N) 单独赋予对 BAY 38-4766 的抗性。两种突变蛋白的组合对耐药性表现出强大的协同作用，重建了体外突变体的高耐药表型。这些发现与 TCRB (2,5,6-三氯-1-β -D-呋喃核糖苯并咪唑)抗性的遗传图谱一致(P.M. Krosky et al., J. Virol. 72:4721-4728, 1998)，并提供了进一步的间接证据，证明由 UL89 和 UL56 编码的蛋白质作为 CMV 终止酶的两个亚基发挥作用。虽然这些研究还表明 BAY 38-4766 的分子机制与苯并咪唑核糖核苷不同，但它们也为 BAY 38-4766 出色的特异性和耐受性提供了解释，因为哺乳动物 DNA 不经历类似的成熟步骤。
3-Hydroxy-2,2-dimethyl-N-[4({[5-(dimethylamino)-1-naphthyl]sulfonyl}amino)-phenyl]propanamide (BAY 38-4766) is a novel selective nonnucleoside inhibitor of cytomegalovirus (CMV) replication with an excellent safety profile. This compound and structural analogues inhibit neither viral DNA synthesis nor viral transcription and translation. Accumulation of dense bodies and noninfectious enveloped particles coincides with inhibition of both concatemer processing and functional cleavage at intergenomic transitions, pointing to interference with viral DNA maturation and packaging of monomeric genome lengths. Resistant virus populations, including a murine CMV (MCMV) isolate with 566-fold-decreased drug sensitivity, were selected in vitro. Sequencing of the six open reading frames (ORFs) known to be essentially involved in viral DNA cleavage and packaging identified mutations in ORFs UL56, UL89, and UL104. Construction of MCMV recombinants expressing different combinations of murine homologues of mutant UL56, UL89, and UL104 and analysis of drug susceptibilities clearly demonstrated that mutant ORFs UL89 exon II (M360I) and M56 (P202A I208N) individually confer resistance to BAY 38-4766. A combination of both mutant proteins exhibited a strong synergistic effect on resistance, reconstituting the high-resistance phenotype of the in vitro mutant. These findings are consistent with genetic mapping of resistance to TCRB (2,5,6-trichloro-1-beta -D-ribofuranosyi benzimidazole) (P.M. Krosky et al., J. Virol. 72:4721-4728, 1998) and provide further indirect evidence that proteins encoded by UL89 and UL56 function as two subunits of the CMV terminase. While these studies also suggest that the molecular mechanism of BAY 38-4766 is distinct from that of benzimidazole ribonucleosides, they also offer an explanation for the excellent specificity and tolerability of BAY 38-4766, since mammalian DNA does not undergo comparable maturation steps.