The activation mechanisms and reactivity of ruthenium metallo-prodrug lead structures were investigated in detail using capillary zone electrophoresis mass spectrometry (CZE–MS) in a time-dependent manner and by exposing to a protein/oligonucleotide mixture. The competitive assays were performed with sodium trans-[RuCl₄(HInd)₂] where Hind = indazole (NKP-1339), [(η⁶-p-cymene)RuCl₂(pta)], where pta = 1,3,5-triaza-7-phosphaadamantane (RAPTA-C) and [(η⁶-biphenyl)RuCl(1,2-ethylenediamine)]PF₆ (RM175). Molecular and quantitative information on binding preferences was obtained by coupling CZE to electrospray ionization MS (ESI-MS) and inductively coupled plasma MS (ICP-MS), respectively. A score system is presented that ranks the binding preferences of Ru complexes with nucleotides and demonstrated the following trend of decreasing selectivity after 24 h: RM175 (0.89) > RAPTA-C (0.78) > NKP-1339 (0.40). As expected, the organometallic drug candidates RM175 and RAPTA-C underwent a halido/aqua ligand exchange reaction at the metal center and showed distinct reactivity towards the biomolecules. In particular, the protein/DNA binding sites of RAPTA-C in a mixture of protein (ubiquitin) and oligonucleotide (5′-dATTGGCAC-3′) were located at single-amino acid and single-nucleotide resolution, respectively. Activated RAPTA-C bound selectively to Met1, adenine and cytosine in this setting, which contrasts with the selectivity of RM175 for guanine. Finally, activation products of NKP-1339 were detected corresponding to Ru^II(Hind)₂ fragments coordinated to the oligonucleotide, which represents one of the few examples of a directly observed Ru^II adduct.

使用毛细管区带电泳质谱法（CZE-MS）以时间依赖的方式并暴露于蛋白质/寡核苷酸混合物中，详细研究了钌金属前药铅结构的活化机理和反应性。在竞争性测定用钠进行反式-将[RuCl ₄（后）₂ ]，其中后腿=吲唑（NKP-1339），[（η ⁶ - p -cymene）的RuCl ₂（PTA）]，其中PTA = 1,3， 5-三氮杂-7-磷（RAPTA-C）和[（η ⁶ -联苯）的RuCl（1,2-乙二胺）] PF ₆（RM175）。通过分别将CZE耦合到电喷雾电离MS（ESI-MS）和电感耦合等离子体MS（ICP-MS），可以获得有关结合偏好的分子和定量信息。提出了评分系统，该评分系统对Ru复合物与核苷酸的结合偏好进行排名，并证明了24小时后选择性降低的以下趋势：RM175（0.89）> RAPTA-C（0.78）> NKP-1339（0.40）。如预期的那样，候选有机金属药物RM175和RAPTA-C在金属中心进行了卤代/水配体交换反应，并显示出对生物分子的独特反应性。特别地，在蛋白质（遍在蛋白）和寡核苷酸（5'-dATTGGCAC-3'）的混合物中，RAPTA-C的蛋白质/ DNA结合位点分别位于单氨基酸和单核苷酸分辨率。在这种情况下，活化的RAPTA-C与Met1，腺嘌呤和胞嘧啶选择性结合，这与RM175对鸟嘌呤的选择性形成对比。最后，检测到与Ru相对应的NKP-1339激活产物^II（Hind）₂片段与寡核苷酸配位，代表直接观察到的Ru ^II加合物的几个例子之一。