Advancing humanized monoclonal antibody for counteracting fentanyl toxicity towards clinical development

Abstract

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.

Antibody-mediated cellular responses are dysregulated in Multisystem Inflammatory Syndrome in Children (MIS-C)

“…It was expressed recombinantly on a human IgG1/IgKappa scaffold or as a bispecific killer engager (BiKE) containing the α-CD235a as a single-chain variable fragment (scFv) fused to a nanobody specific for human CD16 (Felices et al, 2020; Vallera et al, 2020). The anti-CD235a hIgG1 reagent was cloned, expressed, and purified following the previously described recombinant antibody production process (Hicks et al, 2022). VH and VL gene fragments for Gibson assembly were codon optimized and synthesized by Twist Bioscience.…”

Nanotherapeutics for Alleviating Anesthesia‐Associated Complications

“…[ 123 ] Murine, chimeric and humanized antibodies have been developed to capture or catalyze fentanyl and its analogs to counteract opioid toxicity. [ 124 , 125 , 126 ] For instance, a humanized monoclonal antibody, C10‐S66K displayed a high binding capacity to fentanyl and carfentanil (100‐fold more potent than fentanyl). Intraperitoneal injection of 30 mg kg −1 C10‐S66K antibody significantly alleviated respiratory depression caused by carfentanil overdose ( Figure 3 a ).…”

Mechanisms of Neurorespiratory Toxicity Induced by Fentanyl Analogs—Lessons from Animal Studies

“…The initial fentanyl-induced drop in ventilation could also rapidly (3 min) return to 50% of baseline ventilation if fentanyl was administered to a person pretreated 24 h before with a 500 mg dose of fentanyl antibodies. Novel murine-derived anti-fentanyl mAbs were selected thereafter based on their high affinity to fentanyl (binding association rate constant of >1 nmoL −1 ·s −1 and dissociation constant of <0.7 h −1 ), which could predict their efficacy to block fentanyl-induced effects in mice [ 80 ]. Currently, at least two humanized anti-fentanyl mAbs deserve clinical testing to assess their potential to treat fentanyl toxicity.…”