Combatting Synthetic Designer Opioids: A Conjugate Vaccine Ablates Lethal Doses of Fentanyl Class Drugs

Abstract

Fentanyl is an addictive prescription opioid that is over 80 times more potent than morphine.The synthetic nature of fentanyl has enabled the creation of dangerous "designer drug" analogues that escape toxicology screening,y et display comparable potency to the parent drug. Alarmingly,alarge number of fatalities have been linked to overdose of fentanyl derivatives.H erein, we report an effective immunotherapyfor reducing the psychoactive effects of fentanyl class drugs.A single conjugate vaccine was created that elicited high levels of antibodies with cross-reactivity for aw ide panel of fentanyl analogues.M oreover,v accinated mice gained significant protection from lethal fentanyl doses.Lastly,asurface plasmon resonance (SPR)-based technique was established enabling drug-specificity profiling of antibodies derived directly from serum. Our newly developed fentanyl vaccine and analytical methods mayassist in the battle against synthetic opioid abuse. Figure 5. Antiserum opioid binding curves and SPR sensorgrams. a) Diluted mouse serum from week-6 was incubated with serial dilutions of the listed opioids and injected into aBiacore 3000 containing aF ent-BSA-loaded sensor chip. Signal produced by antibody binding to the SPR chip without drug present was used as areference for 100 %b inding. Fentanyls used were racemic and 3-Me was cis.b )Overlaid plots of sensorgrams obtained for the interaction between fentanyl (1000, 500, 250, 125, 62.5, 31.25, 15.63, 7.81, 3.9, 1.95, and 0nm)a nd immobilized anti-fentanyl antibodiesa t258 8ConaBiOptix 404pi. Original experimental sensorgrams are shown in black and fitted curves are traced in white.

Vaccine blunts fentanyl potency in male rhesus monkeys

“…Fentanyl vaccines have been developed and evaluated in both mice and rats (Bremer et al, 2016;Raleigh et al, 2019;Torten et al, 1975). For example, the fentanyltetanus toxoid (TT) conjugate vaccine used in the present study produced a 33-fold antinociceptive potency shift for fentanyl and a 9-fold shift antinociceptive potency for amethylfentanyl in mice (Bremer et al, 2016). In addition, a fentanyl-keyhole limpet hemocyanin (KLH) conjugate vaccine produced a 5.4-fold antinociceptive potency shift for fentanyl in rats (Raleigh et al, 2019).…”

“…The fentanyl vaccine was composed of a fentanyl hapten conjugated to tetanus toxoid (TT) as described previously (Bremer et al, 2016) and solubilized in 50% glycerol and 50% phosphate-buffered saline. Fentanyl copies were 23 per protein, based on conjugation with surrogate bovine serum albumin.…”

“…Preclinical research is a critical component in the development of novel therapeutics to address the opioid crisis. Fentanyl vaccines have been developed and evaluated in both mice and rats (Bremer et al, 2016;Raleigh et al, 2019;Torten et al, 1975). For example, the fentanyltetanus toxoid (TT) conjugate vaccine used in the present study produced a 33-fold antinociceptive potency shift for fentanyl and a 9-fold shift antinociceptive potency for amethylfentanyl in mice (Bremer et al, 2016).…”

Vaccines to combat the opioid crisis

“…Fentanyl and its analogs are commonly used as adulterants in heroin and account for a recent spike in overdose deaths. Our laboratory has developed the first vaccine against fentanyl, a TT conjugate with alum and CpG ODN adjuvants, which generates antibodies with single-digit nanomolar affinities, shifted the fentanyl dose-response curve 30-fold in antinociception testing in mice, and protected not only against fentanyl, but commonly observed fentanyl ester analogs [8]. Most importantly, the vaccine was able to protect against lethal doses of 2.2 and 4.4 mg/kg; unvaccinated mice experienced 18 and 55% overdose fatalities at these doses.…”

Studies towards the improvement of an anti-cocaine monoclonal antibody for treatment of acute overdose

“…A cocaine-BSA conjugate was immobilized on the chip and samples from each time point were passed through the flow cell. 14 The GNCgzk scFv-Fc exhibited an adequate serum half-life of 3.9 days (Figure 2) in mice, an improvement over the GNC92H2 counterpart. 7 …”