The effects of a repeated dose of a recombinant humanized anti-cocaine monoclonal antibody on cocaine self-administration in rats

Wetzel, Hanna N.; Tsibulsky, Vladimir L.; Norman, Andrew B.

doi:10.1016/j.drugalcdep.2016.09.024

Cited by 19 publications

(18 citation statements)

References 29 publications

(40 reference statements)

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“…As potential immunotherapeutics for the treatment of cocaine abuse, the murine-derived chimeric mAb GNC92H2 (Carrera et al, 2005), the humanized mAb h2E2 (Norman et al, 2014) and the fully human anti-cocaine mAb GNCgzk (Treweek & Janda, 2012) have been generated and characterized with regard to their affinities and specificities for cocaine and its metabolites. Additionally, in rodent models the in vivo ability of anti-cocaine mAbs to (i) decrease brain cocaine concentrations by restricting its distribution from plasma, (ii) increase the drug levels needed to reinstate cocaine-dependent self-administration behavior in trained rats and (iii) substantially reduce cocaine toxicity/lethality have been well documented (Treweek & Janda, 2012;Norman et al, 2007Norman et al, , 2009Norman & Ball, 2012;Wetzel et al, 2016). Based on such preliminary studies, the mAb h2E2 is entering phase 1 clinical trials for cocaine abuse therapy.…”

Section: Introductionmentioning

confidence: 99%

Structural analysis of free and liganded forms of the Fab fragment of a high-affinity anti-cocaine antibody, h2E2

et al. 2019

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A high-affinity anti-cocaine monoclonal antibody, designated h2E2, is entering phase 1 clinical trials for cocaine abuse therapy. To gain insight into the molecular details of its structure that are important for binding cocaine and cocaine metabolites, the Fab fragment was generated and crystallized with and without ligand. Structures of the unliganded Fab and the Fab fragment bound to benzoylecgonine were determined, and were compared with each other and with other crystallized anti-cocaine antibodies. The affinity of the h2E2 antibody for cocaine is 4 nM, while that of the cocaine metabolite benzoylecgonine is 20 nM. Both are higher than the reported affinity for cocaine of the two previously crystallized anti-cocaine antibodies. Consistent with cocaine fluorescent quenching binding studies for the h2E2 mAb, four aromatic residues in the CDR regions of the Fab (TyrL32, TyrL96, TrpL91 and TrpH33) were found to be involved in ligand binding. The aromatic side chains surround and trap the tropane moiety of the ligand in the complex structure, forming significant van der Waals interactions which may account for the higher affinity observed for the h2E2 antibody. A water molecule mediates hydrogen bonding between the antibody and the carbonyl group of the benzoyl ester. The affinity of binding to h2E2 of benzoylecgonine differs only by a factor of five compared with that of cocaine; therefore, it is suggested that h2E2 would bind cocaine in the same way as observed in the Fab–benzoylecgonine complex, with minor rearrangements of some hypervariable segments of the antibody.

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Section: Introductionmentioning

confidence: 99%

Structural analysis of free and liganded forms of the Fab fragment of a high-affinity anti-cocaine antibody, h2E2

et al. 2019

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“…It has a long half-life in both rats and mice [5,7]. This mAb also increased the amount of cocaine needed to reinstate self-administration behavior in a rat model of relapse by 3-fold [8], which should translate into a decrease in the probability of cocaine induced relapse. On the basis of these findings h2E2 is a lead candidate as an immunotherapeutic for cocaine abuse.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a recombinant humanized anti-cocaine monoclonal antibody produced from multiple clones for the selection of a master cell bank candidate

Wetzel

Webster

Saeed

et al. 2017

Biochemical and Biophysical Research Communications

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We have generated a humanized anti-cocaine monoclonal antibody (mAb), which is at an advanced stage of pre-clinical development. We report here in vitro binding affinity studies, and in vivo pharmacokinetic and efficacy studies of the recombinant mAb. The overall aim was to characterize the recombinant antibody from each of the three highest producing transfected clones and to select one to establish a master cell bank. In mAb pharmacokinetic studies, after injection with h2E2 (120 mg/kg iv) blood was collected from the tail tip of mice over 28 days. Antibody concentrations were quantified using ELISA. The h2E2 concentration as a function of time was fit using a two-compartment pharmacokinetic model. To test in vivo efficacy, mice were injected with h2E2 (120 mg/kg iv), then one hour later injected with an equimolar dose of cocaine. Blood and brain were collected 5 minutes after cocaine administration. Cocaine concentrations were quantified using LC/MS. The affinity of the antibody for cocaine was determined using a [3H] cocaine binding assay. All three antibodies had long elimination half-lives, 2–5 nM Kds for cocaine, and prevented cocaine’s entry into the brain by sequestering it in the plasma. Pharmacokinetic and radio-ligand binding assays supported designation of the highest producing clone (85) as the master cell bank candidate. Overall, the recombinant h2E2 showed favorable binding properties, pharmacokinetics, and in vivo efficacy.

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“…The mAb h2E2 has high affinity and selectivity for cocaine over its inactive metabolites [4,5] and has a long plasma half-life in both rats and mice. In rats, the recombinant humanized mAb has been shown to decrease brain concentrations of cocaine [4] and prevent cocaine-induced priming of self-administration behavior [6]. In mice, h2E2 can antagonize the distribution of cocaine's active metabolite, cocaethylene, into the brain [7].…”

Section: Introductionmentioning

confidence: 99%

“…However, many of these assumptions have been called into question. For example, it is speculated that high selectivity of the humanized anti-cocaine mAb h2E2 over the major inactive metabolite benzoylecgonine may not be ideal, and may exacerbate the increase in cocaine consumption that is observed after the mAb has been surmounted [6]. Extensive study of anti-methamphetamine mAbs with varying pharmacological profiles has also revealed that in vitro affinities and in vivo pharmacokinetic half-life are not always predictive of the magnitude and duration of action of these antibodies [8].…”

Section: Introductionmentioning

confidence: 99%

A mathematical model of a recombinant humanized anti-cocaine monoclonal antibody's effects on cocaine pharmacokinetics in mice

2017

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Aims A recombinant humanized anti-cocaine monoclonal antibody (mAb), h2E2, is at an advanced stage of pre-clinical development as an immunotherapy for cocaine abuse. It is hypothesized that h2E2 binds to and sequesters cocaine in the blood. Main Methods A three-compartment model of the effects of h2E2 on cocaine's distribution was constructed. The model assumes that h2E2 binds to cocaine and that the h2E2-cocaine complex does not enter the brain but distributes between the central and peripheral compartments. Free cocaine is eliminated from both the central and peripheral compartments, and h2E2 and the h2E2-cocaine complex are eliminated from the central compartment only. This model was tested against a new dataset measuring cocaine concentrations in the brain and plasma over one hour in the presence and absence of h2E2. Key findings The mAb significantly increased plasma cocaine concentrations with a concomitant significant decrease in brain concentration. Plasma concentrations declined over the 1-hour sampling period in both groups. With a set of parameters within reasonable physiological ranges, the three-compartment model was able to qualitatively and quantitatively simulate the increased plasma concentration in the presence of the antibody and the decreased peak brain concentration in the presence of antibody. Importantly, the model explained the decline in plasma concentrations over time as distribution of the cocaine-h2E2 complex into a peripheral compartment. Significance This model will facilitate the targeting of ideal mAb PK/PD properties thus accelerating the identification of lead candidate anti-drug mAbs.

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The effects of a repeated dose of a recombinant humanized anti-cocaine monoclonal antibody on cocaine self-administration in rats

Cited by 19 publications

References 29 publications

Structural analysis of free and liganded forms of the Fab fragment of a high-affinity anti-cocaine antibody, h2E2

Structural analysis of free and liganded forms of the Fab fragment of a high-affinity anti-cocaine antibody, h2E2

Characterization of a recombinant humanized anti-cocaine monoclonal antibody produced from multiple clones for the selection of a master cell bank candidate

A mathematical model of a recombinant humanized anti-cocaine monoclonal antibody's effects on cocaine pharmacokinetics in mice

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