Effects of a cocaine hydrolase engineered from human butyrylcholinesterase on metabolic profile of cocaine in rats.
Accelerating cocaine metabolism through enzymatic hydrolysis at cocaine benzoyl ester is recognized as a promising therapeutic approach for cocaine abuse treatment. Our more recently designed A199S/F227A/S287G/A328W/Y332G mutant of human BChE, denoted as cocaine hydrolase-3 (CocH3), has a considerably improved catalytic efficiency against cocaine and has been proven active in blocking cocaine-induced toxicity and physiological effects. In the present study, we have further characterized the effects of CocH3 on the detailed metabolic profile of cocaine in rats administrated intravenously (IV) with 5 mg/kg cocaine, demonstrating that IV administration of 0.15 mg/kg CocH3 dramatically changed the metabolic profile of cocaine. Without CocH3 administration, the dominant cocaine-metabolizing pathway in rats was cocaine methyl ester hydrolysis to benzoylecgonine (BZE). With the CocH3 administration, the dominant cocaine-metabolizing pathway in rats became cocaine benzoyl ester hydrolysis to ecgonine methyl ester (EME), and the other two metabolic pathways (i.e. cocaine methyl ester hydrolysis to BZE and cocaine oxidation to norcocaine) became insignificant. The CocH3-catalyzed cocaine benzoyl ester hydrolysis to EME was so efficient such that the measured maximum blood cocaine concentration (∼38 ng/ml) was significantly lower than the threshold blood cocaine concentration (∼72 ng/ml) required to produce any measurable physiological effects.
['Chen|Xiabin|X|', 'Zheng|Xirong|X|', 'Zhou|Ziyuan|Z|', 'Zhan|Chang-Guo|CG|', 'Zheng|Fang|F|']
D000818:Animals D055162:Biocatalysis D002091:Butyrylcholinesterase D002851:Chromatography, High Pressure Liquid D003042:Cocaine D006801:Humans D006867:Hydrolases D008297:Male D010084:Oxidation-Reduction D015202:Protein Engineering D051381:Rats D017207:Rats, Sprague-Dawley D011994:Recombinant Proteins D053719:Tandem Mass Spectrometry
Cocaine hydrolase Drug abuse Drug metabolism Enzyme Metabolite