Expression and characterization of a humanized cocaine‐binding antibody

Redwan, Elrashdy M.; Larsen, Nicholas; Zhou, Bin; Wirsching, Peter; Janda, Kim D.; Wilson, Ian A.

doi:10.1002/bit.10598

Cited by 22 publications

(15 citation statements)

References 27 publications

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“…Passive immunization with murine anti-cocaine mAbs has also been shown in rats to attenuate the behavioral effects of cocaine (Carrera et al, 1995;Fox et al, 1996;Mets et al, 1998;Carrera et al, 2000) and therefore represents a potential adjunct to active immunization (Kosten and Owens, 2005), or an emergency rescue treatment in instances of cocaine overdose. However, for optimal safety and efficacy in clinical use, anti-cocaine mAbs should have a human sequence and structure (Redwan et al, 2003;Norman and Ball, 2012).…”

Section: Introductionmentioning

confidence: 99%

A Recombinant Humanized Anti-Cocaine Monoclonal Antibody Inhibits the Distribution of Cocaine to the Brain in Rats

Norman¹,

Gooden²,

Tabet³

et al. 2014

Drug Metab Dispos

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The monoclonal antibody (mAb), h2E2, is a humanized version of the chimeric human/murine anti-cocaine mAb 2E2. The recombinant h2E2 protein was produced in vitro from a transfected mammalian cell line and retained high affinity (4 nM K d ) and specificity for cocaine over its inactive metabolites benzoylecgonine (BE) and ecgonine methyl ester. In rats, pharmacokinetic studies of h2E2 (120 mg/kg i.v.) showed a long terminal elimination half-life of 9.0 days and a low volume of distribution at steady state (V dss ) of 0.3 l/kg. Pretreatment with h2E2 produced a dramatic 8.8-fold increase in the area under the plasma cocaine concentration-time curve (AUC) and in brain a concomitant decrease of 68% of cocaine's AUC following an i.v. injection of an equimolar cocaine dose. Sequestration of cocaine in plasma by h2E2, shown via reduction of cocaine's V dss , indicates potential clinical efficacy. Although the binding of cocaine to h2E2 in plasma should inhibit distribution and metabolism, the elimination of cocaine remained multicompartmental and was still rapidly eliminated from plasma despite the presence of h2E2. BE was the major cocaine metabolite, and brain BE concentrations were sixfold higher than in plasma, indicating that cocaine is normally metabolized in the brain. In the presence of h2E2, brain BE concentrations were decreased and plasma BE was increased, consistent with the observed h2E2-induced changes in cocaine disposition. The inhibition of cocaine distribution to the brain confirms the humanized mAb, h2E2, as a lead candidate for development as an immunotherapy for cocaine abuse.

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Section: Introductionmentioning

confidence: 99%

A Recombinant Humanized Anti-Cocaine Monoclonal Antibody Inhibits the Distribution of Cocaine to the Brain in Rats

Norman¹,

Gooden²,

Tabet³

et al. 2014

Drug Metab Dispos

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“…The expression cassette consisted of cytomegalovirus (CMV)-enhancer chicken b-actin (CAG) promoter, the anti-cocaine IgG1 monoclonal antibody heavy chain with secretion signal and light chain sequence derived from the GNC92H2 Fab separated by a furin 2A self-cleavage site and the rabbit a-globin polyadenylation signal (Fig. 1A) (Niwa et al, 1991;Carrera et al, 2000;Redwan et al, 2003;Fang et al, 2005;De et al, 2006;Mao et al, 2011). The assembled full-length cDNA was sequenced by overlapping PCR to validate that the sequence was without mutations.…”

Section: Aavrh10anticocmabmentioning

confidence: 99%

AAVrh.10-Mediated Expression of an Anti-Cocaine Antibody Mediates Persistent Passive Immunization That Suppresses Cocaine-Induced Behavior

Rosenberg

Hicks

et al. 2012

Human Gene Therapy

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“…It has now been established that mAb 2E2 has a human sequence ␥ 1 heavy and a murine light chain (see Materials and Methods). In addition to 2E2, Redwan et al (2003) have generated, characterized, and "humanized" the murine anticocaine mAb GNC92H2, the murine version of which has been shown to have in vivo efficacy in rat models of cocaine addiction (Carrera et al, 2001). In addition, two catalytic murine anticocaine mAb that are designed to reduce blood cocaine levels through its hydrolysis have been generated and characterized (Matsushita et al, 2001;Larsen et al, 2004).…”

mentioning

confidence: 99%

A Chimeric Human/Murine Anticocaine Monoclonal Antibody Inhibits the Distribution of Cocaine to the Brain in Mice

Norman¹,

Tabet²,

Norman³

et al. 2006

J Pharmacol Exp Ther

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The predominantly human sequence, high-affinity anticocaine monoclonal antibody (mAb) 2E2 was cleared slowly from mouse blood by a first-order process with an elimination t 1/2 of 8.1 days. Infused 2E2 also produced a dramatic dose-dependent increase in plasma cocaine concentrations and a concomitant decrease in the brain cocaine concentrations produced by an i.v. injection of cocaine HCl (0.56 mg/kg). At the highest dose of 2E2 tested (3:1, mAb/drug), cocaine was not detectable in the brain. Pharmacokinetic studies showed that the normal disappearance of cocaine from plasma was described by a two-compartment pharmacokinetic model with distribution t 1/2␣ and terminal elimination t 1/2␤ values of 1.9 and 26.1 min, respectively. In the presence of an equimolar dose of mAb 2E2, there was a 26-fold increase in the area under the plasma cocaine concentration-time curve (AUC) relative to the AUC in the absence of 2E2. Consequently, 2E2 decreased the volume of distribution of cocaine from 6.0 to 0.20 l/kg, which approximated that of 2E2 (0.28 l/kg). However, cocaine was still rapidly cleared from plasma, and its elimination was now described by a single-compartment model with an elimination t 1/2 of 17 min. Importantly, 2E2 also produced a 4.5-fold (78%) decrease in the cocaine AUC in the brain. Therefore, the effect of 2E2 on plasma and brain cocaine concentrations was predominantly caused by a change in the distribution of cocaine with negligible effects on its rate of clearance. These data support the concept of immunotherapy for drug abuse.Despite decades of basic and clinical research there is still no approved pharmacotherapy for the prevention of relapse in cocaine abusers (Vocci and Ling, 2005). The drug-induced reinstatement (priming) of drug self-administration behavior represents an animal model of relapse (DeWitt and Stewart, 1981) with the concentration of cocaine in the body a critical determinant of the probability of reinstating cocaine selfadministration (Norman et al., 1999(Norman et al., , 2002. Because the site of action of cocaine is presumably in the brain, decreasing the concentrations reaching the brain would be expected to decrease the probability of relapse. Antibodies with high affinity and specificity for cocaine would be expected to act as pharmacokinetic antagonists by sequestering cocaine in the peripheral circulation and preventing its entry to the brain. Indeed, active immunization of animals with hapten-carrier conjugates can elicit the production of polyclonal anticocaine antibodies with sufficient levels and affinity for cocaine that they can reduce the amount of cocaine entering the brain (Fox et al., 1996). Active immunization has also been shown to attenuate the behavioral effects (Carrera et al., 1995;Fox et al., 1996;Ettinger et al., 1997) and the priming effect (Carrera et al., 2000) of systemically administered cocaine in rats. Furthermore, the ability of active immunization to produce levels of polyclonal anticocaine antibodies in humans (Kosten et al., 2002) that we...

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Expression and characterization of a humanized cocaine‐binding antibody

Cited by 22 publications

References 27 publications

A Recombinant Humanized Anti-Cocaine Monoclonal Antibody Inhibits the Distribution of Cocaine to the Brain in Rats

A Recombinant Humanized Anti-Cocaine Monoclonal Antibody Inhibits the Distribution of Cocaine to the Brain in Rats

AAVrh.10-Mediated Expression of an Anti-Cocaine Antibody Mediates Persistent Passive Immunization That Suppresses Cocaine-Induced Behavior

A Chimeric Human/Murine Anticocaine Monoclonal Antibody Inhibits the Distribution of Cocaine to the Brain in Mice

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