Siegert Markus, Kranawetvogl Andreas, Thiermann Horst, John Harald
Department of Chemistry, Humboldt-Universität zu Berlin, Berlin, Germany.
Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany.
Drug Test Anal. 2018 Feb;10(2):243-253. doi: 10.1002/dta.2299. Epub 2017 Nov 23.
The vesicant sulfur mustard (SM) is a banned chemical warfare agent. Although, SM has been used in combat since WWI, there is no causal therapy currently available. Accordingly, development and investigation of antidotes and scavengers targeting SM are of high clinical relevance. N-acetylcysteine (NAC) was shown to mitigate symptoms of SM intoxications in vitro and in vivo. However, it is still unclear whether the beneficial effects of NAC are only due to physiological processes or also due to chemical scavenging of SM. Therefore, in this study, we examined the scavenging potential of NAC toward SM. Co-incubations of SM and different NAC concentrations in human serum were performed to monitor diverse adducts (covalent reaction products) of human serum albumin (HSA), NAC, and SM. After proteolytic cleavage of HSA with proteinase K the alkylated tripeptide hydroxyethylthioethyl-CysProPhe (HETE-CPF) and the disulfide bridged tripeptide NAC-CPF were detected. Samples were analyzed by microbore liquid chromatography-electrospray ionization-high-resolution tandem-mass spectrometry (μLC-ESI MS/HR MS). Furthermore, degradation kinetics of SM in phosphate buffered saline were measured in the presence and absence of NAC. Although NAC-CPF was identified and characterized for the first time by mass spectrometry and reaction products of NAC and SM were detected and identified by MS/HR MS, analyses clearly documented minor reactivity not significantly contributing to reduction of SM concentrations. Therefore, we conclude that chemical scavenging of SM by NAC does not play the key role in NAC therapy of SM poisoning.
糜烂性毒剂硫芥是一种被禁用的化学战剂。尽管自第一次世界大战以来硫芥就被用于战争，但目前尚无特效疗法。因此，开发和研究针对硫芥的解毒剂和清除剂具有很高的临床意义。N-乙酰半胱氨酸（NAC）已被证明在体外和体内均可减轻硫芥中毒的症状。然而，NAC的有益作用是仅归因于生理过程还是也归因于对硫芥的化学清除作用仍不清楚。因此，在本研究中，我们检测了NAC对硫芥的清除潜力。将硫芥与人血清中不同浓度的NAC共同孵育，以监测人血清白蛋白（HSA）、NAC和硫芥的各种加合物（共价反应产物）。用蛋白酶K对HSA进行蛋白水解裂解后，检测到烷基化三肽羟乙基硫代乙基-半胱氨酸-脯氨酸-苯丙氨酸（HETE-CPF）和二硫键桥连三肽NAC-CPF。样品通过微径液相色谱-电喷雾电离-高分辨串联质谱（μLC-ESI MS/HR MS）进行分析。此外，在有无NAC存在的情况下，测定了硫芥在磷酸盐缓冲盐水中的降解动力学。尽管通过质谱首次鉴定并表征了NAC-CPF，且通过MS/HR MS检测并鉴定了NAC与硫芥的反应产物，但分析清楚地表明反应活性较小，对硫芥浓度的降低没有显著贡献。因此，我们得出结论，NAC对硫芥的化学清除在硫芥中毒的NAC治疗中不发挥关键作用。
