Analogues with fluorescent leaving groups for screening and selection of enzymes that efficiently hydrolyze organophosphorus nerve agents

被引:73
作者
Briseño-Roa, L
Hill, J
Notman, S
Sellers, D
Smith, AP
Timperley, CM [1 ]
Wetherell, J
Williams, NH
Williams, GR
Fersht, AR
Griffiths, AD
机构
[1] Def Sci & Technol Lab, Salisbury SP4 0JQ, Wilts, England
[2] MRC, Ctr Prot Engn, Cambridge CB2 2QH, England
[3] MRC, Ctr Mol Biol, Cambridge CB2 2QH, England
基金
英国医学研究理事会;
关键词
D O I
10.1021/jm050518j
中图分类号
R914 [药物化学];
学科分类号
100701 ;
摘要
Enzymes that efficiently hydrolyze highly toxic organophosphorus nerve agents could potentially be used as medical countermeasures. As sufficiently active enzymes are currently unknown, we synthesized twelve fluorogenic analogues of organophosphorus nerve agents with the 3-chloro-7-oxy-4-methylcoumarin leaving group as probes for high-throughput enzyme screening. This set included analogues of the pesticides paraoxon, parathion, and dimefox, and the nerve agents DFP, tabun, sarin, cyclosarin, soman, VX, and Russian-VX. Data from inhibition of acetylcholinesterase, in vivo toxicity tests of a representative analogue (cyclosarin), and kinetic studies with phosphotriesterase (PTE) from Pseudomonas diminuta, and a mammalian serum paraoxonase (PON1), confirmed that the analogues mimic the parent nerve agents effectively. They are suitable tools for high-throughput screens for the directed evolution of efficient nerve agent organophosphatases.
引用
收藏
页码:246 / 255
页数:10
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