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，在白血病细胞中通过新建立的敏感方法在白血病细胞中测量了ARA-C.ARA-CTP的细胞内活性代谢产物，其中25例患有ARA-C或Log-Cacting Ara-C，低或常规剂量的白血病患者BHAC，BHAC。 ARA-CTP浓度因给药方法，剂量和患者而有所不同，并且不能从血浆ARA-C浓度中预测。由于血浆ARA-C的维持对于保留ARA-CTP在细胞中非常重要，因此有效地产生了ARA-C和BHAC的连续输注ARA-CTP。在BHAC治疗中，完全缓解的患者的ARA-CTP量比没有缓解的患者更大，这表明ARA-C将成为BHAC治疗效率的关键参数。骨髓抑制与等离子体ARA-C水平相关，这表明正常的造血干细胞对个体之间对ARA-C的敏感性相似。2。掺入DNA中的ARA-C检测。检测方法fo…建立了更多的r ara-c融合到DNA中。用ARA-C处理后，将DNA与白血病细胞的酸不溶部分分离。将DNA酶促为包括ARA-C在内的核外侧消化。通过高性能液相色谱分离ARA-C，然后进行液化。每个步骤的恢复超过90％。放射免疫分子将使用抗-ARA-C血清确认其ARA-C浓度3。通过新的计算机控制的体外药代动力学模拟系统评估的Cytotp毒素效应。在药代动力学模拟状况与常规培养系统状况之间比较了隔毒性。与模拟的ARA-C输注孵育2、4、8和16小时的细胞系K562的存活率表明ARA-C的细胞毒性是时间依赖性的。相反，在常规培养系统下，未观察到时间依赖性抑制。同样，对二绿色菜素输注0.5、2、4和8小时的模拟表明，多诺比霉素的细胞毒性作用降低了浓度依赖性。