膦酰化肟对乙酰胆碱酯酶的抑制作用及中毒酶重活化特性研究——基于效应标志物质谱定量分析技术

将对氧磷(Paraoxon)与肟类重活化剂双复磷(Obidoxime)反应制得膦酰化肟(DEP-obidoxime);利用基于效应标志物的质谱定量技术研究了膦酰化肟对乙酰胆碱酯酶(AChE)的抑制作用及中毒酶的重活化特性.结果表明,膦酰化肟具有极强的AChE抑制毒性,但因膦酰化肟中毒酶与原有机磷毒物中毒酶结构相同,故其膦酰化AChE仍可被经典重活化剂如氯解磷定(Pralidoxime)、双复磷(Obidoxime)及酰胺磷定(HI-6)]重活化,根据EC_(50)表征结果,这3种重活化剂的重活化效果强弱顺序依次为氯解磷定HI-6双复磷.

Studies on Poisoning Characteristics of Phosphylated Oxime on AChE and Efficacy of Reactivators Based on Quantitative Determination of Effect Biomarkers by Liquid Chromatograph-mass Spectrometry

Organophosphorus compounds(OPs) have been extensively utilized throughout the world for many years, however, the security problems are always concerned due to the high toxicity.OPs primarily inhibit acetylcholinesterase(AChE) in nerve system and will cause cholinergic crisis in the synaptic cleft.Enzyme reactivatorsd representing oxime derivatives of quarternary pyridinium compounds are considered as specific medications.Recently, the exact clinical effectiveness of oxime reactivators is a matter of controversy.Although oxime reactivators can recover the activity of AChE and improve neuromuscular transmission, some study reports found oximes could also react with OPs and formmore potent inhibitors of AChE during the process of reactivation, which may intoxicate the reactivated AChE once again, even worsen the poisoning of enzyme due to the high reactivity.In this paper, paraoxon as an example of OPs and obidoxime representing a typical oxime were chosen to demonstrate the reactivation efficacy of oximes in the treatment of OPs poisoning.The reaction product diethylphosphoryl obidoxime(DEP-obidoxime) was first prepared.Then, the poisoning characteristics of phosphylated oxime on AChE and efficacy of reactivators were studied via quantitative determination of the effect biomarkers formed between paraoxon and AChE.The experimental results confirmed that DEP-obidoxime can intoxicate AChE in extremely short time, fortunately, the poisoned AChE by DEP-obidoxime can still be reactivated by typical oximespralidoxime, pyramidoxime monohydrate(HI-6) and obidoxime].EC50 values of each oxime were determined and indicated that the reactivation effectiveness of the above oximes as the following ranking: pralidoxime>HI-6>obidoxime.