Woods JH

References (10)

Title : Design, preparation, and characterization of high-activity mutants of human butyrylcholinesterase specific for detoxification of cocaine - Xue_2011_Mol.Pharmacol_79_290
Author(s) : Xue L , Ko MC , Tong M , Yang W , Hou S , Fang L , Liu J , Zheng F , Woods JH , Tai HH , Zhan CG
Ref : Molecular Pharmacology , 79 :290 , 2011
Abstract : 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.
ESTHER : Xue_2011_Mol.Pharmacol_79_290
PubMedSearch : Xue_2011_Mol.Pharmacol_79_290
PubMedID: 20971807

Title : Subunit stabilization and polyethylene glycolation of cocaine esterase improves in vivo residence time - Narasimhan_2011_Mol.Pharmacol_80_1056
Author(s) : Narasimhan D , Collins GT , Nance MR , Nichols J , Edwald E , Chan J , Ko MC , Woods JH , Tesmer JJ , Sunahara RK
Ref : Molecular Pharmacology , 80 :1056 , 2011
Abstract : No small-molecule therapeutic is available to treat cocaine addiction, but enzyme-based therapy to accelerate cocaine hydrolysis in serum has gained momentum. Bacterial cocaine esterase (CocE) is the fastest known native enzyme that hydrolyzes cocaine. However, its lability at 37 degrees C has limited its therapeutic potential. Cross-linking subunits through disulfide bridging is commonly used to stabilize multimeric enzymes. Herein we use structural methods to guide the introduction of two cysteine residues within dimer interface of CocE to facilitate intermolecular disulfide bond formation. The disulfide-crosslinked enzyme displays improved thermostability, particularly when combined with previously described mutations that enhance stability (T172R-G173Q). The newly modified enzyme yielded an extremely stable form of CocE (CCRQ-CocE) that retained greater than 90% of its activity after 41 days at 37 degrees C, representing an improvement of more than 4700-fold over the wild-type enzyme. CCRQ-CocE could also be modified by polyethylene glycol (PEG) polymers, which improved its in vivo residence time from 24 to 72 h, as measured by a cocaine lethality assay, by self-administration in rodents, and by measurement of inhibition of cocaine-induced cardiovascular effects in rhesus monkeys. PEG-CCRQ elicited negligible immune response in rodents. Subunit stabilization and PEGylation has thus produced a potential protein therapeutic with markedly higher stability both in vitro and in vivo.
ESTHER : Narasimhan_2011_Mol.Pharmacol_80_1056
PubMedSearch : Narasimhan_2011_Mol.Pharmacol_80_1056
PubMedID: 21890748
Gene_locus related to this paper: rhosm-cocE

Title : Structural analysis of thermostabilizing mutations of cocaine esterase - Narasimhan_2010_Protein.Eng.Des.Sel_23_537
Author(s) : Narasimhan D , Nance MR , Gao D , Ko MC , MacDonald J , Tamburi P , Yoon D , Landry DM , Woods JH , Zhan CG , Tesmer JJ , Sunahara RK
Ref : Protein Engineering Des Sel , 23 :537 , 2010
Abstract : Cocaine is considered to be the most addictive of all substances of abuse and mediates its effects by inhibiting monoamine transporters, primarily the dopamine transporters. There are currently no small molecules that can be used to combat its toxic and addictive properties, in part because of the difficulty of developing compounds that inhibit cocaine binding without having intrinsic effects on dopamine transport. Most of the effective cocaine inhibitors also display addictive properties. We have recently reported the use of cocaine esterase (CocE) to accelerate the removal of systemic cocaine and to prevent cocaine-induced lethality. However, wild-type CocE is relatively unstable at physiological temperatures (tau(1/2) approximately 13 min at 37 degrees C), presenting challenges for its development as a viable therapeutic agent. We applied computational approaches to predict mutations to stabilize CocE and showed that several of these have increased stability both in vitro and in vivo, with the most efficacious mutant (T172R/G173Q) extending half-life up to 370 min. Here we present novel X-ray crystallographic data on these mutants that provide a plausible model for the observed enhanced stability. We also more extensively characterize the previously reported variants and report on a new stabilizing mutant, L169K. The improved stability of these engineered CocE enzymes will have a profound influence on the use of this protein to combat cocaine-induced toxicity and addiction in humans.
ESTHER : Narasimhan_2010_Protein.Eng.Des.Sel_23_537
PubMedSearch : Narasimhan_2010_Protein.Eng.Des.Sel_23_537
PubMedID: 20436035
Gene_locus related to this paper: rhosm-cocE

Title : A thermally stable form of bacterial cocaine esterase: a potential therapeutic agent for treatment of cocaine abuse - Brim_2010_Mol.Pharmacol_77_593
Author(s) : Brim RL , Nance MR , Youngstrom DW , Narasimhan D , Zhan CG , Tesmer JJ , Sunahara RK , Woods JH
Ref : Molecular Pharmacology , 77 :593 , 2010
Abstract : Rhodococcal cocaine esterase (CocE) is an attractive potential treatment for both cocaine overdose and cocaine addiction. CocE directly degrades cocaine into inactive products, whereas traditional small-molecule approaches require blockade of the inhibitory action of cocaine on a diverse array of monoamine transporters and ion channels. The usefulness of wild-type (wt) cocaine esterase is hampered by its inactivation at 37 degrees C. Herein, we characterize the most thermostable form of this enzyme to date, CocE-L169K/G173Q. In vitro kinetic analyses reveal that CocE-L169K/G173Q displays a half-life of 2.9 days at 37 degrees C, which represents a 340-fold improvement over wt and is 15-fold greater than previously reported mutants. Crystallographic analyses of CocE-L169K/G173Q, determined at 1.6-A resolution, suggest that stabilization involves enhanced domain-domain interactions involving van der Waals interactions and hydrogen bonding. In vivo rodent studies reveal that intravenous pretreatment with CocE-L169K/G173Q in mice provides protection from cocaine-induced lethality for longer time periods before cocaine administration than wt CocE. Furthermore, intravenous administration (pretreatment) of CocE-L169K/G173Q prevents self-administration of cocaine in a time-dependent manner. Termination of the in vivo effects of CoCE seems to be dependent on, but not proportional to, its clearance from plasma as its half-life is approximately 2.3 h and similar to that of wt CocE (2.2 h). Taken together these data suggest that CocE-L169K/G173Q possesses many of the properties of a biological therapeutic for treating cocaine abuse but requires additional development to improve its serum half-life.
ESTHER : Brim_2010_Mol.Pharmacol_77_593
PubMedSearch : Brim_2010_Mol.Pharmacol_77_593
PubMedID: 20086035
Gene_locus related to this paper: rhosm-cocE

Title : Thermostable variants of cocaine esterase for long-time protection against cocaine toxicity - Gao_2009_Mol.Pharmacol_75_318
Author(s) : Gao D , Narasimhan DL , MacDonald J , Brim R , Ko MC , Landry DW , Woods JH , Sunahara RK , Zhan CG
Ref : Molecular Pharmacology , 75 :318 , 2009
Abstract : Enhancing cocaine metabolism by administration of cocaine esterase (CocE) has been recognized as a promising treatment strategy for cocaine overdose and addiction, because CocE is the most efficient native enzyme for metabolizing the naturally occurring cocaine yet identified. A major obstacle to the clinical application of CocE is the thermoinstability of native CocE with a half-life of only a few minutes at physiological temperature (37 degrees C). Here we report thermostable variants of CocE developed through rational design using a novel computational approach followed by in vitro and in vivo studies. This integrated computational-experimental effort has yielded a CocE variant with a approximately 30-fold increase in plasma half-life both in vitro and in vivo. The novel design strategy can be used to develop thermostable mutants of any protein.
ESTHER : Gao_2009_Mol.Pharmacol_75_318
PubMedSearch : Gao_2009_Mol.Pharmacol_75_318
PubMedID: 18987161

Title : Most efficient cocaine hydrolase designed by virtual screening of transition states - Zheng_2008_J.Am.Chem.Soc_130_12148
Author(s) : Zheng F , Yang W , Ko MC , Liu J , Cho H , Gao D , Tong M , Tai HH , Woods JH , Zhan CG
Ref : Journal of the American Chemical Society , 130 :12148 , 2008
Abstract : Cocaine is recognized as the most reinforcing of all drugs of abuse. There is no anticocaine medication available. The disastrous medical and social consequences of cocaine addiction have made the development of an anticocaine medication a high priority. It has been recognized that an ideal anticocaine medication is one that accelerates cocaine metabolism producing biologically inactive metabolites via a route similar to the primary cocaine-metabolizing pathway, i.e., cocaine hydrolysis catalyzed by plasma enzyme butyrylcholinesterase (BChE). However, wild-type BChE has a low catalytic efficiency against the abused cocaine. Design of a high-activity enzyme mutant is extremely challenging, particularly when the chemical reaction process is rate-determining for the enzymatic reaction. Here we report the design and discovery of a high-activity mutant of human BChE by using a novel, systematic computational design approach based on transition-state simulations and activation energy calculations. The novel computational design approach has led to discovery of the most efficient cocaine hydrolase, i.e., a human BChE mutant with an approximately 2000-fold improved catalytic efficiency, promising for therapeutic treatment of cocaine overdose and addiction as an exogenous enzyme in human. The encouraging discovery resulted from the computational design not only provides a promising anticocaine medication but also demonstrates that the novel, generally applicable computational design approach is promising for rational enzyme redesign and drug discovery.
ESTHER : Zheng_2008_J.Am.Chem.Soc_130_12148
PubMedSearch : Zheng_2008_J.Am.Chem.Soc_130_12148
PubMedID: 18710224

Title : Dopamine agonist-induced yawning in rats: a dopamine D3 receptor-mediated behavior - Collins_2005_J.Pharmacol.Exp.Ther_314_310
Author(s) : Collins GT , Witkin JM , Newman AH , Svensson KA , Grundt P , Cao J , Woods JH
Ref : Journal of Pharmacology & Experimental Therapeutics , 314 :310 , 2005
Abstract : A specific role for the dopamine D3 receptor in behavior has yet to be elucidated. We now report that dopamine D2/D3 agonists elicit dose-dependent yawning behavior in rats, resulting in an inverted U-shaped dose-response curve. A series of experiments was directed toward the hypothesis that the induction of yawning is a D3 receptor-mediated effect, whereas the inhibition of the yawning observed at higher doses is due to competing D2 receptor activity. We compared several dopaminergic agonists with a range of in vitro D3 selectivity, including PD-128,907 [(S)-(+)-(4aR, 10bR)-3,4,4a,10b-tetrahydro-4-propyl-2H,5H-[1]benzopyrano-[4,3-b]-1,4-oxazin-9-ol HCl], PD-128,908 [(R)-(-)-(4aS,10bS)-3,4,4a,10b-tetrahydro-4-propyl-2H,5H-[1]benzopyrano-[4,3-b]-1 ,4-oxazin-9-ol HCl], quinelorane [(5aR-trans)-5,5a,6,7,8, 9,9a,10-octahydro-6-propylpyrido[2,3-g]quinazolin-2-amine dihydrochloride], pramipexole (N'-propyl-4,5,6,7-tetrahydrobenzothiazole-2,6-diamine), 7-OH-DPAT [(+/-)-7-hydroxy-2-dipropylaminotetralin HBr], quinpirole [trans-(-)-(4aR)-4,4a,5,6,7,8, 8a,9-octahydro-5-propyl-1H-pyrazolo[3,4-g]quinoline HCl], bromocriptine [(+)-2-bromo-12'-hydroxy-2'-(1-methylethyl)-5'-(2-methylpropyl) ergotaman-3',6'-18-trione methanesulfonate], and apomorphine [(R)-(-)-5,6,6a,7-tetrahydro-6-methyl-4H-dibenzo-[de,g]quinoline-10,11-diol HCl] with respect to their ability to induce yawning in rats. A series of D2/D3 antagonists differing in selectivity for D3 over D2 receptors were evaluated for their ability to alter the effects of the dopamine agonists. The antagonists L-741,626 (3-[4-(4-chlorophenyl)-4-hydroxypiperidin-l-yl]methyl-1H-indole), haloperidol (4-[4-(4-chlorophenyl)-4-hydroxy-1-piperidinyl]-1-(4-fluorophenyl)-1-butanone HCl), nafadotride (N-[(1-butyl-2-pyrrolidinyl)methyl]-4-cyano-1-methoxy-2-naphtha-lenecarboxamide), U99194 (2,3-dihydro-5,6-dimethoxy-N,N-dipropyl-1H-inden-2-amine maleate), SB-277011A (trans-N-[4-[2-(6-cyano-1,2,3,4-tetrahydroisoquinolin-2-yl)ethyl]cyclohexyl]-4-qu inolinecarboxamide), and PG01037 (N-{4-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-trans-but-2-enyl}-4-pyridine-2-yl-b enzamide HCl) were used to determine effects on dose-response curves for D2/D3 agonist-induced yawning. In addition, the potential contribution of cholinergic and/or serotonergic mechanisms to the yawning response was investigated using a series of pharmacological tools including scopolamine [(a,S)-a-(hydroxymethyl)benzeneacetic acid (1a,2b,4b,5a,7b)-9-methyl-3-oxa-9-azatricyclo[3.3.1.02,4]-non7-yl ester hydrobromide], mianserin (1,2,3,4,10,14b-hexahydro-2-methyldibenzo[c,f]pyrazino[1,2-a]azepine HCl), and the D3-preferring antagonists nafadotride, U99194, SB-277011A, and PG01037 to differentially modulate yawning induced by PD-128,907, physostigmine [(3aS)-cis-1,2,3,3a,8,8a-hexahydro-1,3a,8-trimethylpyrrolo[2,3-b]indol-5-ol methylcarbamate hemisulfate], and N-[3-(trifluoromethyl)phenyl]piperazine HCl. The results of these experiments provide convergent evidence that dopamine D2/D3 agonist-induced yawning is a D3 agonist-mediated behavior, with subsequent inhibition of yawning being driven by competing D2 agonist activity. Thus, dopamine agonist-induced yawning may represent an in vivo method for selectively identifying D3 and D2 receptor-mediated activities.
ESTHER : Collins_2005_J.Pharmacol.Exp.Ther_314_310
PubMedSearch : Collins_2005_J.Pharmacol.Exp.Ther_314_310
PubMedID: 15833897

Title : Large scale purification of butyrylcholinesterase from human plasma suitable for injection into monkeys\; A potential new therapeutic for protection against cocaine end nerve agent toxicity - Lockridge_2005_J.Med.Chem.Biol.Radiol.Def_3_nihms5095
Author(s) : Lockridge O , Schopfer LM , Winger G , Woods JH
Ref : Journal of Medicinal Chemistryical Biology Radiol Def , 3 :nihms5095 , 2005
Abstract : Pretreatment of animals with butyrylcholinesterase (EC 3.1.1.8 BChE) provides complete protection from the acute effects of organophosphorus nerve agents. Butyrylcholinesterase has also been shown to protect from cocaine toxicity. Large amounts of highly purified butyrylcholinesterase are needed to test the effectiveness of this new therapeutic agent in monkeys. Only a minimum amount of endotoxin can be present in a therapeutic intended for injection into monkeys. Our goal was to develop a large scale purification method for human BChE from human plasma with precautions to minimize endotoxin content. A protocol was developed that processed up to 100 L of human plasma at a time. Dialysis in pH 4.0 buffer, ion exchange chromatography at pH 4, affinity chromatography on procainamide-Sepharose, and HPLC ion exchange at pH 7.4 yielded highly purified human BChE containing a low endotoxin level of about 800 EU/ml. The purified BChE produced by this method had a mean residence time of 56 h in mice and 93 h in monkeys, and caused no toxic effects. The absence of a toxic effect in monkeys demonstrates that the endotoxin level of 800 EU/ml was well tolerated by monkeys.
ESTHER : Lockridge_2005_J.Med.Chem.Biol.Radiol.Def_3_nihms5095
PubMedSearch : Lockridge_2005_J.Med.Chem.Biol.Radiol.Def_3_nihms5095
PubMedID: 16788731

Title : Characterization of butyrylcholinesterase antagonism of cocaine-induced hyperactivity - Koetzner_2002_Drug.Metab.Dispos_30_716
Author(s) : Koetzner L , Woods JH
Ref : Drug Metabolism & Disposition: The Biological Fate of Chemicals , 30 :716 , 2002
Abstract : Although there are several published demonstrations that exogenous butyrylcholinesterase (EC 3.1.1.8) works to antagonize cocaine in vivo, a systematic characterization of the enzyme-drug interaction is lacking as is confirmation of the mechanism of effect. This has been addressed using cocaine-induced locomotor activity in mice as a behavioral endpoint. The enzyme was effective, but the enzyme dose-antagonist effect relationship revealed an asymptotic partial maximum effect. This effect was not due to dose-dependent enzyme pharmacokinetics or to a stimulant effect of the cocaine metabolites but rather to partial metabolism of cocaine. Since neither metabolite of cocaine inhibited enzyme activity as potently as cocaine, partial metabolism is not likely due to end-product inhibition. The enzyme reduced the maximum effect of cocaine on locomotor activity. The mechanistic data are generally consistent: the enzyme was inactive against the nonester dopamine/norepinephrine uptake inhibitor, nomifensine, and a paraoxon-inactivated sample of enzyme was ineffective. However, the enzyme was effective against bupropion, a nonester dopamine uptake inhibitor.
ESTHER : Koetzner_2002_Drug.Metab.Dispos_30_716
PubMedSearch : Koetzner_2002_Drug.Metab.Dispos_30_716
PubMedID: 12019200

Title : Characterization of equine butyrylcholinesterase disposition in the mouse - Koetzner_2002_Drug.Metab.Dispos_30_724
Author(s) : Koetzner L , Woods JH
Ref : Drug Metabolism & Disposition: The Biological Fate of Chemicals , 30 :724 , 2002
Abstract : Butyrylcholinesterase administration has been shown to block the effects of cocaine. However, even in model systems, the pharmacokinetics of the enzyme are only partly understood. Measurements of plasma enzyme concentration, antibody titer determinations, and measurement of cocaine-induced locomotor activity in mice were used to describe the disposition of butyrylcholinesterase. Clearance of the enzyme showed biexponential kinetics; the first component was sensitive to asialofetuin, suggesting a role for the asialoglycoprotein receptor. Cocaine did not influence enzyme disposition. An antibody response to enzyme injection was seen; the role of this response is not clear. The antagonist effect of the enzyme was eliminated faster than the enzyme was eliminated from plasma; this may be due to a contribution of tissue esterases to cocaine metabolism. Intraperitoneal enzyme administration was not effective against cocaine, suggesting that the utility of the enzyme is route-dependent.
ESTHER : Koetzner_2002_Drug.Metab.Dispos_30_724
PubMedSearch : Koetzner_2002_Drug.Metab.Dispos_30_724
PubMedID: 12019201