PT - JOURNAL ARTICLE AU - Liu Xue AU - Mei-Chuan Ko AU - Min Tong AU - Wenchao Yang AU - Shurong Hou AU - Lei Fang AU - Junjun Liu AU - Fang Zheng AU - James H. Woods AU - Hsin-Hsiung Tai AU - Chang-Guo Zhan TI - Design, Preparation, and Characterization of High-Activity Mutants of Human Butyrylcholinesterase Specific for Detoxification of Cocaine AID - 10.1124/mol.110.068494 DP - 2011 Feb 01 TA - Molecular Pharmacology PG - 290--297 VI - 79 IP - 2 4099 - http://molpharm.aspetjournals.org/content/79/2/290.short 4100 - http://molpharm.aspetjournals.org/content/79/2/290.full SO - Mol Pharmacol2011 Feb 01; 79 AB - Cocaine is a widely abused drug without a U.S. Food and Drug Administration-approved medication. There is a recognized, promising anticocaine medication to accelerate cocaine metabolism, producing biologically inactive metabolites via a route similar to the primary cocaine-metabolizing pathway [i.e., cocaine hydrolysis catalyzed by butyrylcholinesterase (BChE) in plasma]. An ideal, therapeutically valuable mutant of human BChE should have not only a significantly improved catalytic activity against (−)-cocaine but also certain selectivity for (−)-cocaine over neurotransmitter acetylcholine (ACh), such that one would not expect systemic administration of the BChE mutant to interrupt cholinergic transmission. The present study accounting for the mutation-caused changes of the catalytic activities of BChE against both (−)-cocaine and ACh by means of molecular modeling and site-directed mutagenesis has led to identification of three BChE mutants that have not only a considerably improved catalytic efficiency against (−)-cocaine but also the desirable selectivity for (−)-cocaine over ACh. Two representative BChE mutants have been confirmed to be potent in actual protection of mice from acute toxicity (convulsion and lethality) of a lethal dose of cocaine (180 mg/kg). Pretreatment with the BChE mutant (i.e., 1 min before cocaine administration) dose-dependently protected mice against cocaine-induced convulsions and lethality. In particular, all mice pretreated with the mutant (e.g., 0.02 mg or more of A199S/F227A/S287G/A328W/E441D BChE) survived. The in vivo data reveal the primary factor (i.e., the relative catalytic efficiency), determining the efficacy in practical protection of mice from the acute cocaine toxicity and future direction for further improving the efficacy of the enzyme in the cocaine overdose treatment.