Prediction of anticancer effect of 1-p-D-arabinofuranosylcytostee by sensitive monitoring of its intracellular active metabolite in leukemic cells

通过灵敏监测白血病细胞内的活性代谢物来预测 1-p-D-arabinofuranosylcytostee 的抗癌作用

基本信息

批准号：
10670938
负责人：
UEDA Takanori
金额：
$ 1.02万
依托单位：
Fukui Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2001
项目状态：
已结题

项目摘要

1. Pharmacokinetic study of ara-CTP, an intracellular active metabolite of ara-C.ara-CTP was measured in leukemic cells by the newly established sensitive method in 25 leukemic patients receiving ara-C or log-acting ara-C, BHAC at low or conventional doses. The ara-CTP concentrations differed by the administration methods, doses, and patients, and were not predicted from the plasma ara-C concentrations. As the maintenance of the plasma ara-C was important for the retention of ara-CTP in the cell, continuous infusion of ara-C and BHAC produced ara-CTP efficiently. In BHAC therapy, patients with complete remission achieved greater ara-CTP amounts than those without remission, suggesting that ara-CTP would 6e a crucial parameter for the therapeutic efficacy of BHAC. Myelosuppression was correlated to the plasma ara-C level, suggesting that normal hematopoietic stem cells have similar sensitivity to ara-C among individuals.2. Detection of ara-C incorporated into DNA.The detection method fo … More r ara-C incorporated into DNA was established. DNA was separated from acid insoluble fraction of leukemic cells after treatment with ara-C. The DNA was digested enzymatically to nucleosides that included ara-C. ara-C was isolated by high performance liquid chromatography, followed by liophilization. The recovery of each step was over 90 %. Radioimmunoassay will be applied to the isolated sample using anti-ara-C serum to confirm its ara-C concentration.3. Cytotpxic effects evaluated by a new computer-controlled in vitro pharmacokinetic simulation system.Cytotoxicity was compared between a pharmacokinetically simulated condition and a conventional culture system condition. The survival rates of the cell line K562 incubated with the simulated ara-C infusions for 2, 4, 8, and 16 h demonstrated that the cytotoxicity of ara-C was time-dependent. In contrast, under a conventional culture system, no time-dependent inhibition was observed. Similarly, the simulations of the infusion of daunorubicin for 0.5, 2, 4, and 8 h revealed that the cytotoxic effect of daunorubicin was concentration-dependent Less

1. ara-CTP 的药代动力学研究，ara-CTP 是 ara-C 的细胞内活性代谢物。通过新建立的敏感方法，在 25 名接受 ara-C 或对数作用 ara-C、BHAC 的白血病患者中测量了 ara-CTP 在白血病细胞中的药代动力学研究。低剂量或常规剂量的 ara-CTP 浓度因给药方法、剂量和患者而异，并且不能根据血浆 ara-C 浓度进行预测。 ara-C 对于 ara-CTP 在细胞中的保留很重要，连续输注 ara-C 和 BHAC 可以有效地产生 ara-CTP。在 BHAC 治疗中，完全缓解的患者比未缓解的患者获得了更高的 ara-CTP 量，这表明。 ara-CTP 6e 是 BHAC 骨髓抑制治疗效果的关键参数，与血浆 ara-C 水平相关，表明正常造血干细胞对 BHAC 具有相似的敏感性。 2. ara-C 掺入 DNA 的检测建立了用 ara-C 处理后的白血病细胞中 ara-C 掺入 DNA 的检测方法。通过高效液相色谱将DNA酶消化为包含ara-C的核苷，然后冻干。每个步骤的回收率超过90%。使用抗ara-C血清对分离的样品进行放射免疫测定，以确认其ara-C浓度，并通过新的计算机控制的体外药代动力学模拟系统评估细胞毒性。3．细胞系 K562 与模拟 ara-C 输液孵育 2、4、8 和 16 小时的存活率表明，细胞毒性相反，在常规培养系统下，没有观察到时间依赖性抑制，同样，柔红霉素输注0.5、2、4和8小时的模拟表明，ara-C的细胞毒性作用。柔红霉素具有浓度依赖性更少