Intracellular metabolism of the antiherpes agent (S)-1-[3-hydroxy-2-(phosphonylmethoxy)propyl]cytosine.
Intracellular metabolism of the antiherpes agent (S)-1-[3-hydroxy-2-(phosphonylmethoxy)propyl]cytosine.
复制标题
抗疱疹药物 (S)-1-[3-羟基-2-(膦酰甲氧基)丙基]胞嘧啶的细胞内代谢。
DOI：
10.1002/9780470114735.hawley08755
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发表时间：
1992
影响因子：
3.6
通讯作者：
M. Hitchcock
中科院分区：
文献类型：
--
作者：
H. Ho;K. Woods;J. Bronson;H. Boeck;John C. Martin;M. Hitchcock
（S）-1-[3-羟基-2-（膦酰基甲氧基）丙基]胞嘧啶（HPMPC）是一种抗病毒的膦酸酯核苷酸类似物，其显示出对一系列疱疹病毒的活性。[14 C] HPMPC处理细胞的60%甲醇提取物的阴离子交换高效液相色谱分析显示形成了三种主要代谢产物。其中两种被鉴定为磷酸化形式的HPMPC，HPMPC磷酸盐和HPMPC二磷酸盐，通过酸消化后释放HPMPC并在高效液相色谱上与合成标准品共洗脱。第三种代谢产物对碱性磷酸酶裂解具有抗性，但对磷酸二酯酶敏感，拟定为HPMPC磷酸盐加合物。在单纯疱疹病毒-1感染的细胞中检测到相同的三种代谢物，其浓度与未感染细胞中的浓度相当。当从培养基中去除HPMPC时，细胞中代谢物的浓度缓慢降低，HPMPC磷酸盐、HPMPC二磷酸盐和HPMPC磷酸盐加合物的半衰期分别约为6、17和48小时。HPMPC二磷酸抑制单纯疱疹病毒-1和-2 DNA聚合酶，Ki低于DNA聚合酶α，在每种情况下酶抑制是竞争性的。HPMPC磷酸盐在细胞中的形成和持久性以及HPMPC二磷酸盐对病毒DNA聚合酶的选择性抑制可以解释为什么用HPMPC预处理的细胞即使在HPMPC从培养基中除去后很长时间仍对病毒感染具有抗性。
(S)-1-[3-Hydroxy-2-(phosphonylmethoxy)propyl]cytosine (HPMPC) is an antiviral phosphonate nucleotide analogue that displays activity against a range of herpesviruses. Anion exchange high performance liquid chromatography analysis of the 60% methanol extract from [14C]HPMPC-treated cells reveals the formation of three major metabolites. Two of these were identified as phosphorylated forms of HPMPC, HPMPC phosphate, and HPMPC diphosphate, by liberation of HPMPC upon acid digestion and coelution with synthetic standards on high performance liquid chromatography. The third metabolite, which is resistant to alkaline phosphatase cleavage but sensitive to phosphodiesterase, is proposed to be an HPMPC phosphate adduct. In herpes simplex virus-1-infected cells the same three metabolites are detected, at concentrations comparable to those in uninfected cells. When HPMPC is removed from the medium, the concentrations of the metabolites in cells decrease slowly, with half-lives of approximately 6, 17, and 48 hr for HPMPC phosphate, HPMPC diphosphate, and the HPMPC phosphate adduct, respectively. HPMPC diphosphate inhibits herpes simplex virus-1 and -2 DNA polymerases with a lower Ki than that for DNA polymerase alpha, and enzyme inhibition is competitive in each case. The formation and the persistence of HPMPC phosphates in cells and the selective inhibition of viral DNA polymerases by HPMPC diphosphate can explain why cells pretreated with HPMPC remain refractory to viral infection even long after HPMPC is removed from the medium.