Vol. 302, Issue 2, 710-716, August 2002

Cocaine Metabolism Accelerated by a Re-Engineered Human Butyrylcholinesterase

Hong Sun , Maryann L. Shen, Yuan-Ping Pang , Oksana Lockridge and Stephen Brimijoin

Molecular Neuroscience Program (H.S., S.B.), Department of Molecular Pharmacology and Experimental Therapeutics (H.S., Y.-P.P., S.B.), and Mayo Clinic Cancer Center (Y.-P.P.), Mayo Foundation for Medical Education and Research, Rochester, Minnesota; Eppley Institute and Department of Biochemistry and Molecular Neuroscience (O.L.), University of Nebraska Medical Center, Omaha, Nebraska; and Bruker Daltonics (M.L.S.), Billerica, Massachusetts

Plasma butyrylcholinesterase (BChE) is important in the metabolism of cocaine, but natural human BChE has limited therapeutic potential for detoxication because of low catalytic efficiency with cocaine. Here we report pharmacokinetics of cocaine in rats treated with A328W/Y332A BChE, an excellent cocaine hydrolase designed with the aid of molecular modeling. Compared with wild-type BChE, this enzyme hydrolyzes cocaine with 40-fold improved k_cat (154 min¹ versus 4.1 min¹) and only slightly increased K_M (18 µM versus 4.5 µM). In rats given this hydrolase (3 mg/kg i.v.) 10 min before cocaine challenge (6.8 mg/kg i.v.), cocaine half-life was reduced from 52 min to 18 min. Mirroring the reductions of plasma cocaine were large increases in benzoic acid, a product of BChE-mediated cocaine hydrolysis. All other pharmacokinetic parameters confirmed a large, dose-dependent acceleration of cocaine removal by the injected cocaine hydrolase. These results show that A328W/Y332A, an efficient cocaine hydrolase in vivo as well as in vitro, might promote cocaine detoxication in a clinical setting.