title: The fate of bacterial cocaine esterase (CocE): an in vivo study of CocE-mediated cocaine hydrolysis, CocE pharmacokinetics, and CocE elimination.
abstract: Cocaine abuse and toxicity remain widespread problems in the United States. Currently cocaine toxicity is treated only symptomatically, because there is no Food and Drug Administration-approved pharmacotherapy for this indication. To address the unmet need, a stabilized mutant of bacterial cocaine esterase [T172R/G173Q-CocE (DM-CocE)], which hydrolyzes cocaine into inactive metabolites and has low immunogenic potential, has been developed and previously tested in animal models of cocaine toxicity. Here, we document the rapid cocaine hydrolysis by low doses of DM-CocE in vitro and in vivo, as well as the pharmacokinetics and distribution of the DM-CocE protein in rats. DM-CocE at 50.5 μg/kg effectively eliminated 4 mg/kg cocaine within 2 min in both male and female rats as measured by mass spectrometry. We expanded on these findings by using a pharmacologically relevant dose of DM-CocE (0.32 mg/kg) in rats and monkeys to hydrolyze convulsant doses of cocaine. DM-CocE reduced cocaine to below detection limits rapidly after injection; however, elimination of DM-CocE resulted in peripheral cocaine redistribution by 30 to 60 min. Elimination of DM-CocE was quantified by using [³⁵S] labeling of the enzyme and was found to have a half-life of 2.1 h in rats. Minor urinary output of DM-CocE was also observed. Immunohistochemistry, Western blotting, and radiography all were used to elucidate the mechanism of DM-CocE elimination, rapid proteolysis, and recycling of amino acids into all tissues. This rapid elimination of DM-CocE is a desirable property of a therapeutic for cocaine toxicity and should reduce the likelihood of immunogenic or adverse reactions as DM-CocE moves toward clinical use.
authors: Department of Pharmacology, University of Michigan, Ann Arbor, Michigan, USA.
authors: Remy L
authors: RL
authors: Brim
authors: Kathleen R
authors: KR
authors: Noon
authors: Gregory T
authors: GT
authors: Collins
authors: Aron
authors: A
authors: Stein
authors: Joseph
authors: J
authors: Nichols
authors: Diwa
authors: D
authors: Narasimhan
authors: Mei-Chuan
authors: MC
authors: Ko
authors: James H
authors: JH
authors: Woods
authors: Roger K
authors: RK
authors: Sunahara
mesh_terms: D000818:Animals; D019540:Area Under Curve; D001345:Autoradiography; D001419:Bacteria / Q000201:enzymology*; D015153:Blotting, Western; D002138:Calibration; D002265:Carboxylic Ester Hydrolases / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D003042:Cocaine / Q000097:blood / Q000378:metabolism* / Q000493:pharmacokinetics; D004305:Dose-Response Relationship, Drug; D005260:Female; D005919:Glomerular Filtration Rate; D006868:Hydrolysis; D007124:Immunoenzyme Techniques; D007553:Isotope Labeling; D007700:Kinetics; D008253:Macaca mulatta; D008297:Male; D013058:Mass Spectrometry; D009392:Nephrectomy; D051381:Rats; D017207:Rats, Sprague-Dawley; D013045:Species Specificity; D013050:Spectrometry, Fluorescence; D013462:Sulfur Radioisotopes
chemical_list: D013462:Sulfur Radioisotopes; D002265:Carboxylic Ester Hydrolases; C019921:cocaine esterase; D003042:Cocaine