Cellular pharmacology of multi- and duplex drugsconsisting of ethynylcytidine and 5-fluoro-2′-deoxyuridine
Cellular pharmacology of multi- and duplex drugsconsisting of ethynylcytidine and 5-fluoro-2′-deoxyuridine
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DOI：
10.1007/s10637-009-9353-2
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发表时间：
2011-04-01
影响因子：
3.4
通讯作者：
Peters, Godefridus J.
中科院分区：
文献类型：
--
作者：
Bijnsdorp, Irene V.;Schwendener, Reto A.;Peters, Godefridus J.
前药在增加激活和细胞摄取方面比母体药物有优势。多药ETC- l -FdUrd和双药ETC-FdUrd由两种不同的单磷酸核苷组成，5-氟-2'脱氧尿苷（FdUrd）和乙基胞苷（ETC），分别通过甘油脂或磷酸二酯偶联。该研究的目的是确定细胞毒性水平和药物切割模式。此外，我们确定了ETC-L-FdUrd的脂质体制剂是否会改善细胞毒活性和/或切割。采用标准放射性测定法、高效液相色谱法和LC-MS/MS对FM3A/0乳腺癌细胞及其FdUrd耐药变体FM3A/TK-进行了药物作用/裂解研究。ETC-FdUrd在FM3A/0和TK-细胞中分别有活性（IC50分别为2.2 nM和79 nM）。ETC-L-FdUrd活性较低（IC50: FM3A/0为7 nM， FM3A/TK-为4500 nM）。虽然脂质体制剂在FM3A/0细胞中的活性低于ETC-L-FdUrd (IC50:19.3 nM)，但由于胸苷激酶（TK）缺乏而产生的耐药性大大降低。前药对FM3A/0细胞胸苷酸合成酶（TS）的抑制作用为80-90%，对FM3A/TK-细胞的抑制作用为10-50%。FM3A/TK-细胞中几乎检测不到FdUMP。转运体和核苷酸酶/磷酸酶的抑制导致ETC-FdUrd的细胞毒性降低，表明该药物在细胞外裂解为单磷酸盐，培养基中存在FdUrd和ETC证实了这一点。ETC-L-FdUrd和脂质体制剂不受转运蛋白和核苷酸酶/磷酸酶抑制的影响，表明规避了活性转运蛋白。在体内，ETC-FdUrd和ETC-L-FdURd具有口服活性。ETC核苷酸在肿瘤和肝组织中均有积累。当亲脂连接剂与脂质体制剂结合使用时，这些制剂似乎是有效的。
Prodrugs can have the advantage over parent drugs in increased activation and cellular uptake. The multidrug ETC-L-FdUrd and the duplex drug ETC-FdUrd are composed of two different monophosphate-nucleosides, 5-fluoro-2'deoxyuridine (FdUrd) and ethynylcytidine (ETC), coupled via a glycerolipid or phosphodiester, respectively. The aim of the study was to determine cytotoxicity levels and mode of drug cleavage. Moreover, we determined whether a liposomal formulation of ETC-L-FdUrd would improve cytotoxic activity and/or cleavage. Drug effects/cleavage were studied with standard radioactivity assays, HPLC and LC-MS/MS in FM3A/0 mammary cancer cells and their FdUrd resistant variants FM3A/TK-. ETC-FdUrd was active (IC50 of 2.2 and 79 nM) in FM3A/0 and TK- cells, respectively. ETC-L-FdUrd was less active (IC50: 7 nM in FM3A/0 vs 4500 nM in FM3A/TK-). Although the liposomal formulation was less active than ETC-L-FdUrd in FM3A/0 cells (IC50:19.3 nM), resistance due to thymidine kinase (TK) deficiency was greatly reduced. The prodrugs inhibited thymidylate synthase (TS) in FM3A/0 cells (80-90%), but to a lower extent in FM3A/TK- (10-50%). FdUMP was hardly detected in FM3A/TK- cells. Inhibition of the transporters and nucleotidases/phosphatases resulted in a reduction of cytotoxicity of ETC-FdUrd, indicating that this drug was cleaved outside the cells to the monophosphates, which was verified by the presence of FdUrd and ETC in the medium. ETC-L-FdUrd and the liposomal formulation were neither affected by transporter nor nucleotidase/phosphatase inhibition, indicating circumvention of active transporters. In vivo, ETC-FdUrd and ETC-L-FdURd were orally active. ETC nucleotides accumulated in both tumor and liver tissues. These formulations seem to be effective when a lipophilic linker is used combined with a liposomal formulation.