Efficacy of the rePON1 mutant IIG1 to prevent cyclosarin toxicity in vivo and to detoxify structurally different nerve agents in vitro.
The potent human toxicity of organophosphorus (OP) nerve agents calls for the development of effective antidotes. Standard treatment for nerve agent poisoning with atropine and an oxime has a limited efficacy. An alternative approach is the development of catalytic bioscavengers using OP-hydrolyzing enzymes such as paraoxonases (PON1). Recently, a chimeric PON1 mutant, IIG1, was engineered toward the hydrolysis of the toxic isomers of soman and cyclosarin with high in vitro catalytic efficiency. In order to investigate the suitability of IIG1 as a catalytic bioscavenger, an in vivo guinea pig model was established to determine the protective effect of IIG1 against the highly toxic nerve agent cyclosarin. Prophylactic i.v. injection of IIG1 (1 mg/kg) prevented systemic toxicity in cyclosarin (~2LD50)-poisoned guinea pigs, preserved brain acetylcholinesterase (AChE) activity, and protected erythrocyte AChE activity partially. A lower IIG1 dose (0.2 mg/kg) already prevented mortality and reduced systemic toxicity. IIG1 exhibited a high catalytic efficiency with a homologous series of alkylmethylfluorophosphonates but had low efficiency with the phosphoramidate tabun and was virtually ineffective with the nerve agent VX. This quantitative analysis validated the model for predicting in vivo protection by catalytic bioscavengers based on their catalytic efficiency, the level of circulating enzyme, and the dose of the intoxicating nerve agent. The in vitro and in vivo results indicate that IIG1 may be considered as a promising candidate bioscavenger to protect against the toxic effects of a range of highly toxic nerve agents.
Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, 80937, Munich, Germany, franzworek@bundeswehr.org.
Franz
F
Worek
Thomas
T
Seeger
Moshe
M
Goldsmith
Yacov
Y
Ashani
Haim
H
Leader
Joel S
JS
Sussman
Dan
D
Tawfik
Horst
H
Thiermann
Timo
T
Wille
D000110:Acetylcholinesterase / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D000931:Antidotes / Q000008:administration & dosage / Q000494:pharmacology*; D043303:Aryldialkylphosphatase / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology*; D001921:Brain / Q000187:drug effects / Q000378:metabolism; D002619:Chemical Warfare Agents / Q000633:toxicity*; D002800:Cholinesterase Inhibitors / Q000008:administration & dosage / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D006168:Guinea Pigs; D007275:Injections, Intravenous; D008297:Male; D010755:Organophosphates / Q000008:administration & dosage / Q000633:toxicity; D009943:Organophosphorus Compounds / Q000008:administration & dosage / Q000633:toxicity*; D009946:Organothiophosphorus Compounds / Q000008:administration & dosage / Q000633:toxicity
D000110:Acetylcholinesterase; D000931:Antidotes; D043303:Aryldialkylphosphatase; D002619:Chemical Warfare Agents; D002800:Cholinesterase Inhibitors; D010755:Organophosphates; D009943:Organophosphorus Compounds; D009946:Organothiophosphorus Compounds; C009680:VX-agent; C009374:tabun; C053914:cyclohexyl methylphosphonofluoridate