DNA crosslinking and biological activity of a hairpin polyamide-chlorambucil conjugate
DNA crosslinking and biological activity of a hairpin polyamide-chlorambucil conjugate
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DOI：
10.1093/nar/gkg215
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发表时间：
2003-02-15
影响因子：
14.9
通讯作者：
Beerman, TA
中科院分区：
文献类型：
--
作者：
Wang, YD;Dziegielewski, J;Beerman, TA
一类新型 DNA 烷化剂的原型将 DNA 交联部分苯丁酸氮芥 (Chl) 与序列选择性发夹吡咯-咪唑聚酰胺 ImPy-beta-ImPy-gamma-ImPy-beta-Dp（聚酰胺 1）相结合，对其损伤 DNA 和诱导生物反应的能力进行了评估。聚酰胺 1-Chl 缀合物 (1-Chl) 在无细胞系统中烷基化和链间交联 DNA。烷基化主要发生在 5'-AGCTGCA-3' 序列处，该序列代表聚酰胺结合位点。首先在用 0.1 muM 1-Chl 处理 1 小时的 DNA 上检测到缀合物诱导的损伤，表明缀合物的效力比 Chl 至少强 100 倍。即使在 0.01 muM 浓度下，延长孵育时间也能检测 DNA 损伤。在猿猴病毒 40 (SV40) 体外复制测定中，用 1-Chl 处理可降低 DNA 模板活性。 1-Chl 抑制哺乳动物细胞生长、基因组 DNA 复制和细胞周期进程，并将细胞阻滞在 G(2)/M 期。此外，在 1-Chl 浓度下观察到的细胞效应与无细胞系统中 DNA 损伤所需的浓度相似。母体化合物（未缀合的 Chl 或聚酰胺 1）在相同浓度范围内均未表现出任何细胞活性。缀合物分子 1-Chl 具有聚酰胺的序列选择性和 Chl 增强的 DNA 反应性。
A prototype of a novel class of DNA alkylating agents, which combines the DNA crosslinking moiety chlorambucil (Chl) with a sequence-selective hairpin pyrrole-imidazole polyamide ImPy-beta-ImPy-gamma-ImPy-beta-Dp (polyamide 1), was evaluated for its ability to damage DNA and induce biological responses. Polyamide 1-Chl conjugate (1-Chl) alkylates and interstrand crosslinks DNA in cell-free systems. The alkylation occurs predominantly at 5'-AGCTGCA-3' sequence, which represents the polyamide binding site. Conjugate-induced lesions were first detected on DNA treated for 1 h with 0.1 muM 1-Chl, indicating that the conjugate is at least 100-fold more potent than Chl. Prolonged incubation allowed for DNA damage detection even at 0.01 muM concentration. Treatment with 1-Chl decreased DNA template activity in simian virus 40 (SV40) in vitro replication assays. 1-Chl inhibited mammalian cell growth, genomic DNA replication and cell cycle progression, and arrested cells in the G(2)/M phase. Moreover, cellular effects were observed at 1-Chl concentrations similar to those needed for DNA damage in cell-free systems. Neither of the parent compounds, unconjugated Chl or polyamide 1, demonstrated any cellular activity in the same concentration range. The conjugate molecule 1-Chl possesses the sequence-selectivity of a polyamide and the enhanced DNA reactivity of Chl.