Separation and characterization of oxaliplatin dinucleotides from DNA using HPLC-ESI ion trap mass spectrometry
Separation and characterization of oxaliplatin dinucleotides from DNA using HPLC-ESI ion trap mass spectrometry
复制标题
DOI：
10.1007/s00216-008-2311-6
复制
发表时间：
2008-08
影响因子：
4.3
通讯作者：
S. Mowaka;M. Linscheid
中科院分区：
文献类型：
--
作者：
S. Mowaka;M. Linscheid
奥沙利铂是第三代铂配合物，具有广谱抗肿瘤活性。这种具有DACH载体配体的铂络合物最近受到越来越多的关注，因为它们显示出对顺铂抗性细胞系的功效。由于铂类药物抗肿瘤活性的最重要指标是与基因组DNA形成加合物，因此小牛胸腺DNA-奥沙利铂加合物是本研究的主要靶点。小牛胸腺DNA与奥沙利铂孵育，导致形成大量铂-DNA加合物。用酶的组合，即核酸酶、碱性磷酸酶和核酸酶S1，将处理过的DNA消化成二核苷酸。使用高效液相色谱法，我们进行了分离的个别铂-DNA加合物，同时确定使用电喷雾电离离子阱质谱（MS）。发现1，2-链内和1，2-链间交联加合物;然而，链内性质的加合物的丰度比链间交联的加合物的丰度高得多。其中，d（GpG）-奥沙利铂是最丰富的双功能加合物。在较小程度上，也检测到一些单官能加合物。图m/z902的d（GpG）-Pt（DACH）加合物（a）和m/z 886的d（ApG）-Pt（DACH）加合物（B）的三维视图。DACH 1，2-二氨基环己烷、绿磷、氧化还原氧、浅蓝碳、深蓝氮、灰铂
Oxaliplatin is a third-generation platinum complex, and has a broad spectrum of antitumor activity. Such platinum complexes with the DACH carrier ligand have recently received increasing attention since they show efficacy against cisplatin-resistant cell lines. As the foremost indication of antitumor activity of platinum drugs is the formation of adducts with genomic DNA, calf thymus DNA-oxaliplatin adducts were the major target in this study. Calf thymus DNA was incubated with oxaliplatin, resulting in the formation of a large number of platinum-DNA adducts. Treated DNA was digested into the dinucleotides with a combination of enzymes, namely, benzonase, alkaline phosphatase, and nuclease S1. Using a high-performance liquid chromatography, we carried out the separation of individual platinum-DNA adducts which were concurrently identified using electrospray ionization ion trap mass spectrometry (MS). Both 1,2-intrastrand and 1,2-interstrand cross-linked adducts were found; however, those of the intrastrand nature have a considerably higher abundance than those of the interstrand cross-links. Among them, d(GpG)-oxaliplatin was the most abundant bifuctional adduct. To a lesser extent, a few monofunctional adducts were detected as well. MSnexperiments served to ascertain the detailed structures of oxaliplatin adducts of dinucleoside monophosphates and of dinucleotides.FigureThree-dimensional view of the d(GpG)-Pt(DACH) adduct ofm/z902 (a) and of the d(ApG)-Pt(DACH) adduct ofm/z886 (b).DACH1,2-diaminocyclohexane,greenphosphorus,redoxygen,light bluecarbon,dark bluenitrogen,grayplatinum