The
incidence of fatal overdoses has increased worldwide due to the widespread
access to illicit fentanyl and its potent analogues. Vaccines offer
a promising strategy to reduce the prevalence of opioid use disorders
(OUDs) and to prevent toxicity from accidental and deliberate exposure
to fentanyl and its derivatives. This study describes the development
and characterization of vaccine formulations consisting of novel fentanyl-based
haptens conjugated to carrier proteins. Vaccine efficacy was tested
against opioid-induced behavior and toxicity in mice and rats challenged
with fentanyl and its analogues. Prophylactic vaccination reduced
fentanyl- and sufentanil-induced antinociception, respiratory depression,
and bradycardia in mice and rats. Therapeutic vaccination also reduced
fentanyl intravenous self-administration in rats. Because of their
selectivity, vaccines did not interfere with the pharmacological effects
of commonly used anesthetics nor with methadone, naloxone, oxycodone,
or heroin. These preclinical data support the translation of vaccines
as a viable strategy to counteract fentanyl use disorders and toxicity.
Innovative therapies to complement current treatments are needed to curb the growing incidence of fatal overdoses related to synthetic opioids. Murine and chimeric monoclonal antibodies (mAb) specific for fentanyl and its analogs have demonstrated pre-clinical efficacy in preventing and reversing drug-induced toxicity in rodent models. However, mAb-based therapeutics require extensive engineering as well as
in vitro
and
in vivo
characterization to advance to first-in-human clinical trials. Here, novel murine anti-fentanyl mAbs were selected for development based on affinity for fentanyl, and efficacy in counteracting the pharmacological effects of fentanyl in mice. Humanization and evaluation of mutations designed to eliminate predicted post-translational modifications resulted in two humanized mAbs that were effective at preventing fentanyl-induced pharmacological effects in rats. These humanized mAbs showed favorable biophysical properties with respect to aggregation and hydrophobicity by chromatography-based assays, and thermostability by dynamic scanning fluorimetry. These results collectively support that the humanized anti-fentanyl mAbs developed herein warrant further clinical development for treatment of fentanyl toxicity.
Vaccines and mAbs offer promising strategies to treat substance use disorders (SUDs) and prevent overdose. Despite vaccines and mAbs against SUDs demonstrating proof of efficacy, selectivity, and safety in animal models, it is unknown whether the mechanism of action of these immunotherapeutics relies exclusively on the formation of Ab/drug complexes, or also involves Ab-mediated effector functions. Hence, this study tested whether the efficacy of active and passive immunization against drugs of abuse requires phagocytosis, the intact Fc portion of the anti-drug Ab, FcγRs, or the neonatal FcR (FcRn). The efficacy of a lead vaccine against oxycodone was not diminished in mice after depletion of macrophages or granulocytes. Anti-oxycodone F(ab′)2 fragments resulted in lower serum levels of F(ab′)2 compared with intact mAbs, and F(ab′)2s were not as effective as the parent mAbs in reducing distribution of oxycodone to the brain. The efficacy of vaccines and mAbs against oxycodone was preserved in either FcγIII or FcγI–IV ablated mice, suggesting that FcγRs are not required for Ab efficacy. Finally, both active and passive immunization against oxycodone in FcRn−/− mice yielded reduced efficacy compared with wild-type control mice. These data identified a role for FcRn, but not for phagocytosis or Fc-dependent effector functions, in mediating the efficacy of vaccines and mAbs against SUD. This study supports rational design of vaccines and mAbs engineered for maximal neutralization activity and optimal FcRn binding.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.