107 results on '"Pravetoni, M."'
Search Results
2. Pharmacological mechanisms underlying the efficacy of antibodies generated by a vaccine to treat oxycodone use disorder
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Raleigh, M.D., primary, King, S.J., additional, Baruffaldi, F., additional, Saykao, A., additional, Hamid, F.A., additional, Winston, S., additional, LeSage, M.G., additional, Pentel, P.R., additional, and Pravetoni, M., additional
- Published
- 2021
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3. Structurally distinct nicotine immunogens elicit antibodies with non-overlapping specificities
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Pravetoni, M., Keyler, D. E., Pidaparthi, R. R., Carroll, F. I., Runyon, S. P., Murtaugh, M. P., Earley, C. A., and Pentel, P. R.
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- 2012
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4. Vaccination against nicotine alters the distribution of nicotine delivered via cigarette smoke inhalation to rats
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Pravetoni, M., Keyler, D. E., Raleigh, M. D., Harris, A. C., LeSage, M. G., Mattson, C. K., Pettersson, S., and Pentel, P. R.
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- 2011
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5. Behavioral characterization of mice lacking GIRK/Kir3 channel subunits
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Pravetoni, M. and Wickman, K.
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- 2008
6. Formulation and Characterization of Conjugate Vaccines to Reduce Opioid Use Disorders Suitable for Pharmaceutical Manufacturing and Clinical Evaluation
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Baruffaldi, F., primary, Raleigh, M. D., additional, King, S. J., additional, Roslawski, M. J., additional, Birnbaum, A. K., additional, Hassler, C., additional, Carroll, F. I., additional, Runyon, S. P., additional, Winston, S., additional, Pentel, P. R., additional, and Pravetoni, M., additional
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- 2019
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7. Safety and efficacy of an oxycodone vaccine: Addressing some of the unique considerations posed by opioid abuse
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Raleigh, M. D., primary, Peterson, S. J., additional, Laudenbach, M., additional, Baruffaldi, F., additional, Carroll, F. I., additional, Comer, S. D., additional, Navarro, H. A., additional, Langston, T. L., additional, Runyon, S. P., additional, Winston, S., additional, Pravetoni, M., additional, and Pentel, P. R., additional
- Published
- 2017
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8. Hapten-specific naïve B cells are biomarkers of vaccine efficacy against drugs of abuse
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Taylor, J.J., primary, Laudenbach, M., additional, Tucker, A.M., additional, Jenkins, M.K., additional, and Pravetoni, M., additional
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- 2014
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9. Selective Effects of a Morphine Conjugate Vaccine on Heroin and Metabolite Distribution and Heroin-Induced Behaviors in Rats
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Raleigh, M.D., primary, Pravetoni, M., additional, Harris, A.C., additional, Birnbaum, A.K., additional, and Pentel, P.R., additional
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- 2012
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10. Co-administration of morphine and oxycodone vaccines reduces the distribution of 6-monoacetylmorphine and oxycodone to brain in rats
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Pravetoni, M., primary, Raleigh, M.D., additional, Le Naour, M., additional, Tucker, A.M., additional, Harmon, T.M., additional, Jones, J.M., additional, Birnbaum, A.K., additional, Portoghese, P.S., additional, and Pentel, P.R., additional
- Published
- 2012
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11. An Oxycodone Conjugate Vaccine Elicits Drug-Specific Antibodies that Reduce Oxycodone Distribution to Brain and Hot-Plate Analgesia
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Pravetoni, M., primary, Le Naour, M., additional, Harmon, T. M., additional, Tucker, A. M., additional, Portoghese, P. S., additional, and Pentel, P. R., additional
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- 2012
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12. Absence and Rescue of Morphine Withdrawal in GIRK/Kir3 Knock-out Mice
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Cruz, H. G., primary, Berton, F., additional, Sollini, M., additional, Blanchet, C., additional, Pravetoni, M., additional, Wickman, K., additional, and Luscher, C., additional
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- 2008
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13. Expression and Localization of RGS9-2/G 5/R7BP Complex In Vivo Is Set by Dynamic Control of Its Constitutive Degradation by Cellular Cysteine Proteases
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Anderson, G. R., primary, Lujan, R., additional, Semenov, A., additional, Pravetoni, M., additional, Posokhova, E. N., additional, Song, J. H., additional, Uversky, V., additional, Chen, C.-K., additional, Wickman, K., additional, and Martemyanov, K. A., additional
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- 2007
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14. Selective effects of a morphine conjugate vaccine on heroin and metabolite distribution and heroin-induced behaviors in rats.
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Raleigh, M D, Pravetoni, M, Harris, A C, Birnbaum, A K, and Pentel, P R
- Abstract
Morphine conjugate vaccines have effectively reduced behavioral effects of heroin in rodents and primates. To better understand how these effects are mediated, heroin and metabolite distribution studies were performed in rats in the presence and absence of vaccination. In non-vaccinated rats 6-monoacetylmorphine (6-MAM) was the predominant opioid in plasma and brain as early as 1 minute after i.v. administration of heroin and for up to 14 minutes. Vaccination with morphine conjugated to keyhole limpet hemocyanin (M-KLH) elicited high titers and concentrations of antibodies with high affinity for heroin, 6-MAM, and morphine. Four minutes after heroin administration vaccinated rats showed substantial retention of all three opioids in plasma compared to controls and reduced 6-MAM and morphine, but not heroin, distribution to brain. Administration of 6-MAM rather than heroin in M-KLH vaccinated rats showed a similar drug distribution pattern. Vaccination reduced heroin-induced analgesia and blocked heroin-induced locomotor activity throughout 2 weeks of repeated testing. Higher serum opioid-specific antibody concentrations were associated with higher plasma opioid concentrations, lower brain 6-MAM and morphine concentrations, and lower heroin-induced locomotor activity. Serum antibody concentrations over 0.2 mg/ml were associated with substantial effects on these measures. These data support a critical role for 6-MAM in mediating the early effects of i.v. heroin and suggest that reducing 6-MAM concentration in brain is essential to the efficacy of morphine conjugate vaccines.
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- 2013
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15. Safety and efficacy of an oxycodone vaccine: Addressing some of the unique considerations posed by opioid abuse
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Raleigh, M. D., Peterson, S. J., Laudenbach, M., Baruffaldi, F., Carroll, F. I., Comer, Sandra D., Navarro, H. A., Langston, T. L., Runyon, S. P., Winston, S., Pravetoni, M., and Pentel, P. R.
- Subjects
Psychiatry ,Vaccines ,Opioid abuse ,Medicine ,Oxycodone ,3. Good health - Abstract
Among vaccines aimed at treating substance use disorders, those targeting opioids present several unique medication development challenges. 1) Opioid overdose is a common complication of abuse, so it is desirable for an opioid vaccine to block the toxic as well as the addictive effects of opioids. 2) It is important that an opioid vaccine not interfere with the action of opioid antagonists used to reverse opioid overdose or treat addiction. 3) Some opioids are immunosuppressive and chronic ongoing opioid use could interfere with vaccine immunogenicity. 4) Although antibody-bound oxycodone is unable to enter the brain because of its size, it might still be able to activate peripheral opioid receptors. To assess vaccine impact on opioid toxicity, rats vaccinated with oxycodone conjugated to keyhole limpet hemocyanin subunit dimer (OXY-dKLH) adsorbed to alum or controls vaccinated with dKLH were compared with regard to oxycodone-induced hotplate analgesia and oxycodone-induced respiratory depression and bradycardia. Vaccination shifted the dose-response curves to the right, representing protection, for each of these endpoints. Naloxone was equally effective in both OXY-dKLH and control groups, providing complete and rapid reversal of respiratory depression. The administration of a long-acting naltrexone formulation during vaccination did not impair vaccine immunogenicity in mice. Similarly, serum anti-oxycodone antibody titers were not altered by continuous morphine infusion during vaccination compared to opioid-naïve controls. Competitive ELISA assay showed negligible or low affinity of immune antiserum for endogenous opioids or opioid antagonists. In vitro receptor binding assays showed that antibody-bound oxycodone does not activate mu opioid receptors. These data support further study of OXY-dKLH as a potential treatment for oxycodone abuse and suggest that vaccination might also reduce the severity of oxycodone overdose.
16. Monoclonal Antibodies Engineered with Fc Region Mutations to Extend Protection against Fentanyl Toxicity.
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Khaimraj A, Baehr CA, Hicks D, Raleigh MD, and Pravetoni M
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- Animals, Mice, Humans, Analgesics, Opioid, Half-Life, Protein Engineering, Immunoglobulin G immunology, Fentanyl immunology, Receptors, Fc genetics, Receptors, Fc metabolism, Antibodies, Monoclonal, Histocompatibility Antigens Class I genetics, Histocompatibility Antigens Class I immunology, Mutation, Immunoglobulin Fc Fragments genetics, Immunoglobulin Fc Fragments immunology
- Abstract
Fentanyl and other synthetic opioids are the leading cause of drug-related deaths in the United States. mAbs that selectively target fentanyl and fentanyl analogues offer a promising strategy for treating both opioid-related overdoses and opioid use disorders. To increase the duration of efficacy of a candidate mAb against fentanyl, we selected three sets of mutations in the Fc region of an IgG1 anti-fentanyl mAb (HY6-F9DF215, HY6-F9DHS, HY6-F9YTE) to increase binding to the neonatal Fc receptor (FcRn). The mAb mutants were compared against unmodified (wild-type [WT], HY6-F9WT) anti-fentanyl mAb for fentanyl binding, thermal stability, and FcRn affinity in vitro, and for efficacy against fentanyl and mAb half-life in vivo in mice. Biolayer interferometry showed a >10-fold increase in the affinity for recombinant FcRn of the three mutant mAbs compared with HY6-F9WT. During an acute fentanyl challenge in mice, all FcRn-mutated mAbs provided equal protection against fentanyl-induced effects, and all mAbs reduced brain fentanyl levels compared with the saline group. Serum persistence of the mutant mAbs was tested in Tg276 transgenic mice expressing human FcRn. After administration of 40 mg/kg HY6-F9WT, HY6-F9DF215, HY6-F9DHS, and HY6-F9YTE, the mAbs showed half-lives of 6.3, 26.4, 14.7, and 6.9 d, respectively. These data suggest that modification of mAbs against fentanyl to bind to FcRn with higher affinity can increase their half-life relative to WT mAbs while maintaining efficacy against the toxic effects of fentanyl, further supporting their potential role as a therapeutic treatment option for opioid use disorder and overdose., (Copyright © 2024 by The American Association of Immunologists, Inc.)
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- 2024
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17. Identification and biophysical characterization of a novel domain-swapped camelid antibody specific for fentanyl.
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Gallant JP, Hicks D, Shi K, Moeller NH, Hoppe B, Lake EW, Baehr C, Pravetoni M, Aihara H, and LeBeau AM
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- Animals, Humans, Camelidae immunology, Camelids, New World, Analgesics, Opioid chemistry, Analgesics, Opioid pharmacology, Analgesics, Opioid immunology, Fentanyl chemistry, Fentanyl immunology, Single-Domain Antibodies chemistry, Single-Domain Antibodies immunology
- Abstract
Opioid use disorders (OUD) and overdoses are ever-evolving public health threats that continue to grow in incidence and prevalence in the United States and abroad. Current treatments consist of opioid receptor agonists and antagonists, which are safe and effective but still suffer from some limitations. Murine and humanized monoclonal antibodies (mAb) have emerged as an alternative and complementary strategy to reverse and prevent opioid-induced respiratory depression. To explore antibody applications beyond traditional heavy-light chain mAbs, we identified and biophysically characterized a novel single-domain antibody specific for fentanyl from a camelid variable-heavy-heavy (VHH) domain phage display library. Structural data suggested that VHH binding to fentanyl was facilitated by a unique domain-swapped dimerization mechanism, which accompanied a rearrangement of complementarity-determining region loops leading to the formation of a fentanyl-binding pocket. Structure-guided mutagenesis further identified an amino acid substitution that improved the affinity and relaxed the requirement for dimerization of the VHH in fentanyl binding. Our studies demonstrate VHH engagement of an opioid and inform on how to further engineer a VHH for enhanced stability and efficacy, laying the groundwork for exploring the in vivo applications of VHH-based biologics against OUD and overdose., Competing Interests: Conflict of interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: A. M. L, J. P. G, M. P., C. B., and D. H. are co-inventors on a provisional patent application USPTO--231220--09824.412., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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18. ACE2 decoy Fc-fusions and bi-specific killer engager (BiKEs) require Fc engagement for in vivo efficacy against SARS-CoV-2.
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Dick JK, Hicks D, Krishna VD, Sangala JA, Zandstra BT, Baehr C, Verbeek JS, Cragg MS, Cheeran MC, Pravetoni M, and Hart GT
- Abstract
SARS-CoV-2 virus has continued to evolve over time necessitating the adaptation of vaccines to maintain efficacy. Monoclonal antibodies (mAbs) against SARS-CoV-2 were a key line of defense for unvaccinated or immunocompromised individuals. However, these mAbs are now ineffective against current SARS-CoV-2 variants. Here, we tested three aspects of αSARS-CoV-2 therapeutics. First, we tested whether Fc engagement is necessary for in vivo clearance of SARS-CoV-2. Secondly, we tested bi-specific killer engagers (BiKEs) that simultaneously engage SARS-CoV-2 and a specific Fc receptor. Benefits of these engagers include the ease of manufacturing, stability, more cell-specific targeting, and high affinity binding to Fc receptors. Using both mAbs and BiKEs, we found that both neutralization and Fc receptor engagement were necessary for effective SARS-CoV-2 clearance. Thirdly, due to ACE2 being necessary for viral entry, ACE2 will maintain binding to SARS-CoV-2 despite viral evolution. Therefore, we used an ACE2 decoy Fc-fusion or BiKE, instead of an anti-SARS-CoV-2 antibody sequence, as a potential therapeutic that would withstand viral evolution. We found that the ACE2 decoy approach also required Fc receptor engagement and, unlike traditional neutralizing antibodies against specific variants, enabled the clearance of two distinct SARS-CoV-2 variants. These data show the importance of Fc engagement for mAbs, the utility of BiKEs as therapies for infectious disease, and the in vivo effectiveness of the ACE2 decoy approach. With further studies, we predict combining neutralization, the cellular response, and this ACE2 decoy approach will benefit individuals with ineffective antibody levels., Abbreviations: ACE2, scFv, mAb, BiKE, COVID-19, Fc, CD16, CD32b, CD64, d.p.i., Key Points: With equal dosing, both neutralization and Fc engagement are necessary for the optimal efficacy of in vivo antibodies and bi-specific killer engagers (BiKEs) against SARS-CoV-2. BiKEs can clear SARS-CoV-2 virus and protect against severe infection in the hACE2-K18 mouse model. ACE2 decoys as part of Fc-fusions or BiKEs provide in vivo clearance of two disparate SARS-CoV-2 variants.
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- 2024
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19. The fentanyl-specific antibody FenAb024 can shield against carfentanil effects.
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Urban K, Gkeka A, Chandra M, Greiner D, Pollich S, Ruf S, Kelemen Y, Sundermann T, Pravetoni M, Baehr C, Stebbins CE, Papavasiliou FN, and Verdi JP
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- Animals, Mice, Humans, Fentanyl analogs & derivatives, Fentanyl immunology, Analgesics, Opioid, Antibodies, Monoclonal
- Abstract
The surge in opioid-related deaths, driven predominantly by fentanyl and its synthetic derivatives, has become a critical public health concern, which is particularly evident in the United States. While the situation is less severe in Europe, the European Monitoring Centre for Drugs and Drug Addiction reports a rise in drug overdose deaths, with emerging concerns about the impact of fentanyl-related molecules. Synthetic opioids, initially designed for medical use, have infiltrated illicit markets due to their low production costs and high potency, with carfentanil posing additional threats, including potential chemical weaponization. Existing overdose mitigation heavily relies on naloxone, requiring timely intervention and caregiver presence, while therapeutic prevention strategies face many access challenges. To provide an additional treatment option, we propose the use of a fentanyl-specific monoclonal antibody (mAb), as a non-opioid method of prophylaxis against fentanyl and carfentanil. This mAb shows protection from opioid effects in a pre-clinical murine model. mAbs could emerge as a versatile countermeasure in civilian and biodefense settings, offering a novel approach to combat opioid-associated mortality., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Joseph Verdi reports financial support was provided by National Institute on Drug Abuse. Nina Papavasiliou reports financial support was provided by Helmholtz Foundation. Erec Stebbins reports financial support was provided by Helmholtz Foundation. Joseph Verdi reports a relationship with Hepione Therapeutics Inc. that includes: employment and equity or stocks. Nina Papavasiliou reports a relationship with Hepione Therapeutics Inc. that includes: equity or stocks. Erec Stebbins reports a relationship with Hepione Therapeutics Inc. that includes: board membership and equity or stocks. Yosip Kelemen reports a relationship with Hepione Therapeutics Inc. that includes: employment. Joseph Verdi reports a relationship with Panosome GmbH that includes: employment and equity or stocks. Nina Papavasiliou reports a relationship with Panosome GmbH that includes: board membership and equity or stocks. Erec Stebbins reports a relationship with Panosome GmbH that includes: board membership and equity or stocks. Anastasia Gkeka reports a relationship with Panosome GmbH that includes: employment and equity or stocks. Katharina Urban reports a relationship with Panosome GmbH that includes: employment and equity or stocks. Monica Chandra reports a relationship with Panosome GmbH that includes: employment and equity or stocks. Dennis Greiner reports a relationship with Panosome GmbH that includes: employment. Sandra Ruf reports a relationship with Panosome GmbH that includes: equity or stocks. Selina Pollich reports a relationship with Panosome GmbH that includes: employment. Joseph Verdi has patent pending to Hepione Therapeutics Inc. Nina Papavasiliou has patent pending to Hepione Therapeutics Inc. Erec Stebbins has patent pending to Hepione Therapeutics Inc. Yosip Kelemen has patent pending to Hepione Therapeutics Inc. Marco Pravetoni has patent pending to University of Minnesota. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
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- 2024
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20. Antibody-mediated cellular responses are dysregulated in Multisystem Inflammatory Syndrome in Children (MIS-C).
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Dick JK, Sangala JA, Krishna VD, Khaimraj A, Hamel L, Erickson SM, Hicks D, Soigner Y, Covill LE, Johnson A, Ehrhardt MJ, Ernste K, Brodin P, Koup RA, Khaitan A, Baehr C, Thielen BK, Henzler CM, Skipper C, Miller JS, Bryceson YT, Wu J, John CC, Panoskaltsis-Mortari A, Orioles A, Steiner ME, Cheeran MC, Pravetoni M, and Hart GT
- Abstract
Multisystem Inflammatory Syndrome in Children (MIS-C) is a severe complication of SARS-CoV-2 infection characterized by multi-organ involvement and inflammation. Testing of cellular function ex vivo to understand the aberrant immune response in MIS-C is limited. Despite strong antibody production in MIS-C, SARS-CoV-2 nucleic acid testing can remain positive for 4-6 weeks after infection. Therefore, we hypothesized that dysfunctional cell-mediated antibody responses downstream of antibody production may be responsible for delayed clearance of viral products in MIS-C. In MIS-C, monocytes were hyperfunctional for phagocytosis and cytokine production, while natural killer (NK) cells were hypofunctional for both killing and cytokine production. The decreased NK cell cytotoxicity correlated with an NK exhaustion marker signature and systemic IL-6 levels. Potentially providing a therapeutic option, cellular engagers of CD16 and SARS-CoV-2 proteins were found to rescue NK cell function in vitro. Together, our results reveal dysregulation in antibody-mediated cellular responses unique to MIS-C that likely contribute to the immune pathology of this disease.
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- 2024
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21. Evaluating the rate of reversal of fentanyl-induced respiratory depression using a novel long-acting naloxone nanoparticle, cNLX-NP.
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Averick SE, Kassick AJ, Song D, Zhang B, Vigliaturo J, Luengas D, Silva-Ortiz P, Pravetoni M, and Raleigh MD
- Abstract
Introduction: Fentanyl and fentanyl analogs (F/FA) have become increasingly common adulterants in counterfeit prescription pills and illicit street drug mixtures due to their ease of synthesis and exceedingly high potency. The ongoing epidemic of fatal overdoses fueled by F/FA continues to highlight the need for longer-acting therapies than naloxone (NLX), the current gold-standard for reversing opioid overdoses, which shows limited efficacy to prevent renarcotization associated with F/FA toxicity. A novel opioid reversal agent based on covalent naloxone nanoparticles (cNLX-NP) has been shown to blunt fentanyl-induced respiratory depression out to 48 hr, demonstrating its potential therapeutic utility. The purpose of this study was to characterize how rapidly cNLX-NP reverses fentanyl-induced respiratory effects as well as the duration of its protective effects., Methods: Sprague Dawley male rats (n=6/group) were tested on an oximeter for baseline percent arterial oxygen saturation (%SaO
2 ) challenged with 0.1 mg/kg SC fentanyl and 15 min later given 10 mg/kg IM doses of NLX, nalmefene (NLMF), or cNLX-NP and continuously monitored via oximetry for 10 minutes. One week later the experiment was repeated using a 1:1 mixture of NLX:cNLX-NP as the reversal agent in the rats that previously received NLX alone., Results: While both NLX and NLMF rapidly reversed %SaO2 to baseline within 1 min, rats that received cNLX-NP did not return to >90% SaO2 levels until 9 min after administration. Similarly, heart and breath rates returned to baseline within 1 min of treatment with NLX and NLMF but did not return to baseline until 10 minutes after cNLX-NP administration. In contrast, NLX:cNLX-NP reversed all fentanyl-induced respiratory depressive effects within one minute., Discussion: While cNLX-NP alone may not sufficiently reverse F/FA overdose in a timely manner, mixing free NLX with cNLX-NP can provide a mechanism to both rapidly reverse fentanyl-related effects and maintain extended protection against synthetic opioid toxicity. These data support further development of cNLX-NP as a fast-acting and long-lasting antidote to treat F/FA-induced respiratory depression and overdose, and potentially prevent renarcotization in humans., Competing Interests: A patent application was filed by the Allegheny Health Network US-20220152017-A1 for the preparation of covalent naloxone nanoparticles and SA is an inventor. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Averick, Kassick, Song, Zhang, Vigliaturo, Luengas, Silva-Ortiz, Pravetoni and Raleigh.)- Published
- 2024
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22. Multivalent Vaccination Strategies Protect against Exposure to Polydrug Opioid and Stimulant Mixtures in Mice and Rats.
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Song D, Crouse B, Vigliaturo J, Wu MM, Heimisdottir D, Kassick AJ, Averick SE, Raleigh MD, and Pravetoni M
- Abstract
Illicit drug mixtures containing opioids and stimulants have been responsible for the majority of fatal drug overdoses among occasional users, and those with either opioid use disorder (OUD) or substance use disorder (SUD). As a complementary strategy to current pharmacotherapies, active immunization with conjugate vaccines has been proposed as a viable intervention to treat OUD as well as other SUD for which there are either limited or no treatment options. Vaccination against opioids and stimulants could help address the limitations of current medications (e.g., patient access, compliance, misuse liability, and safety) by providing an additional tool to prevent drug misuse and/or overdoses. However, more research is needed to fully understand the potential benefits and limitations of using vaccines to treat SUD and overdose and to inform us on how to deploy this strategy in the field. Previous reports have shown promise by combining two vaccines into bivalent vaccine formulations to concurrently target multiple drugs. Here, multiple individual candidate monovalent vaccines were incrementally combined in multivalent vaccine formulations to simultaneously target fentanyl, carfentanil, oxycodone, heroin, methamphetamine, and their analogs or metabolites. Bi-, tri-, and quadrivalent vaccine formulations induced the formation of independent serum antibody responses against their respective opioid targets and selectively attenuated the distribution of each individual drug to the brain in mice and rats. Results indicate that a single injection of an admixed multivalent vaccine formulation may be more effective than coinjecting multiple monovalent vaccines at multiple sites. Finally, adding a methamphetamine conjugate vaccine to an quadrivalent opioid vaccine in a pentavalent formulation did not interfere with the production of effective antiopioid IgG antibodies. Multivalent vaccines could provide multifaceted, yet selective, protection against polydrug use and exposure., Competing Interests: The authors declare the following competing financial interest(s): M.P. is the co-inventor of patents disclosing haptens, conjugates, and multivalent formulations. M.P. is the co-inventor of patents disclosing fentanyl hapten-conjugates and methods for making and using same. M.P. and S.A. are co-inventors of patents disclosing fentanyl haptens, fentanyl hapten conjugates, and methods for making and using the same. D.S., M.R., and M.P. are co-inventors of patents disclosing the multivalent formulation and the methamphetamine vaccine. All other authors declare no conflict of interest., (© 2023 American Chemical Society.)
- Published
- 2023
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23. The immunological and pharmacokinetic evaluation of Lipid-PLGA hybrid nanoparticle-based oxycodone vaccines.
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Walter DL, Bian Y, Hu H, Hamid FA, Rostamizadeh K, Vigliaturo JR, DeHority R, Ehrich M, Runyon S, Pravetoni M, and Zhang C
- Subjects
- Animals, Lipids chemistry, Mice, Female, Vaccines pharmacokinetics, Vaccines immunology, Vaccines administration & dosage, Mice, Inbred BALB C, Oxycodone pharmacokinetics, Oxycodone immunology, Oxycodone administration & dosage, Oxycodone chemistry, Nanoparticles chemistry, Polylactic Acid-Polyglycolic Acid Copolymer chemistry
- Abstract
The current opioid epidemic is one of the most profound public health crises facing the United States. Despite that it has been under the spotlight for years, available treatments for opioid use disorder (OUD) and overdose are limited to opioid receptor ligands such as the agonist methadone and the overdose reversing drugs such as naloxone. Vaccines are emerging as an alternative strategy to combat OUD and prevent relapse and overdose. Most vaccine candidates consist of a conjugate structure containing the target opioid attached to an immunogenic carrier protein. However, conjugate vaccines have demonstrated some intrinsic shortfalls, such as fast degradation and poor recognition by immune cells. To overcome these challenges, we proposed a lipid-PLGA hybrid nanoparticle (hNP)-based vaccine against oxycodone (OXY), which is one of the most frequently misused opioid analgesics. The hNP-based OXY vaccine exhibited superior immunogenicity and pharmacokinetic efficacy in comparison to its conjugate vaccine counterpart. Specifically, the hNP-based OXY vaccine formulated with subunit keyhole limpet hemocyanin (sKLH) as the carrier protein and aluminum hydroxide (Alum) as the adjuvant (OXY-sKLH-hNP(Alum)) elicited the most potent OXY-specific antibody response in mice. The induced antibodies efficiently bound with OXY molecules in blood and suppressed their entry into the brain. In a following dose-response study, OXY-sKLH-hNP(Alum) equivalent to 60 μg of sKLH was determined to be the most promising OXY vaccine candidate moving forward. This study provides evidence that hybrid nanoparticle-based vaccines may be superior vaccine candidates than conjugate vaccines and will be beneficial in treating those suffering from OUD., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Chenming Zhang, Marco Pravetoni reports financial support was provided by National Institutes of Health. Zhang, Walter, Hu, Pravetoni, and Hamid have filed patent #VTIP 23–084 pending to Virginia Tech. Other authors declare no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
- Published
- 2025
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24. Self-Adjuvanting TLR7/8 Agonist and Fentanyl Hapten Co-Conjugate Achieves Enhanced Protection against Fentanyl Challenge.
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Powers N, Massena C, Crouse B, Smith M, Hicks L, Evans JT, Miller S, Pravetoni M, and Burkhart D
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- Animals, Mice, Toll-Like Receptor 7 agonists, Fentanyl therapeutic use, Adjuvants, Immunologic therapeutic use, Antigens therapeutic use, Haptens, Analgesics, Opioid therapeutic use, Vaccines, Opioid-Related Disorders prevention & control, Opioid-Related Disorders drug therapy
- Abstract
Currently approved pharmacotherapies for opioid use disorders (OUDs) and overdose reversal agents are insufficient to slow the spread of OUDs due to the proliferation of fentanyl. This is evident in the 31% rise in drug overdose deaths from 2019 to 2022, with rates increasing from 21.6 to 28.3 overdoses per 100,000 deaths. Vaccines are a potential alternative or adjunct therapy for the treatment of several substance use disorders (nicotine, cocaine) but have shown limited clinical success due to suboptimal antibody titers. In this study, we demonstrate that coconjugation of a Toll-like receptor 7/8 (TLR7/8) agonist (UM-3006) alongside a fentanyl-based hapten (F
1 ) on the surface of the carrier protein cross-reactive material 197 (CRM) significantly increased generation of high-affinity fentanyl-specific antibodies. This demonstrated enhanced protection against fentanyl challenges relative to an unconjugated (admix) adjuvant control in mice. Inclusion of aluminum hydroxide (alum) adjuvant further increased titers and enhanced protection, as determined by analysis of fentanyl concentration in serum and brain tissue. Collectively, our findings present a promising approach to enhance the efficacy of antiopioid vaccines, underscoring the need for extensive exploration of TLR7/8 agonist conjugates as a compelling strategy to combat opioid use disorders.- Published
- 2023
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25. A TLR7/8 agonist increases efficacy of anti-fentanyl vaccines in rodent and porcine models.
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Crouse B, Miller SM, Muelken P, Hicks L, Vigliaturo JR, Marker CL, Guedes AGP, Pentel PR, Evans JT, LeSage MG, and Pravetoni M
- Abstract
Opioid use disorders (OUD) and overdose are public health threats worldwide. Widespread access to highly potent illicit synthetic opioids such as fentanyl is driving the recent rise in fatal overdoses. Vaccines containing fentanyl-based haptens conjugated to immunogenic carrier proteins offer a long-lasting, safe, and cost-effective strategy to protect individuals from overdose upon accidental or deliberate exposure to fentanyl and its analogs. Prophylactic or therapeutic active immunization with an anti-fentanyl vaccine induces the production of fentanyl-specific antibodies that bind the drug in the blood and prevent its distribution to the brain, which reduces its reinforcing effects and attenuates respiratory depression and bradycardia. To increase the efficacy of a lead anti-fentanyl vaccine, this study tested whether the incorporation of synthetic toll-like receptor (TLR) 4 and TLR7/8 agonists as vaccine adjuvants would increase vaccine efficacy against fentanyl challenge, overdose, and self-administration in either rats or Hanford miniature pigs. Formulation of the vaccine with a nucleolipid TLR7/8 agonist enhanced its immunogenicity and efficacy in preventing fentanyl-induced respiratory depression, analgesia, bradycardia, and self-administration in either rats or mini-pigs. These studies support the use of TLR7/8 adjuvants in vaccine formulations to improve their clinical efficacy against OUD and potentially other substance use disorders (SUD)., (© 2023. The Author(s).)
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- 2023
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26. A lipidated TLR7/8 adjuvant enhances the efficacy of a vaccine against fentanyl in mice.
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Miller SM, Crouse B, Hicks L, Amin H, Cole S, Bazin HG, Burkhart DJ, Pravetoni M, and Evans JT
- Abstract
Opioid use disorders (OUD) and opioid-related fatal overdoses are a public health concern in the United States. Approximately 100,000 fatal opioid-related overdoses occurred annually from mid-2020 to the present, the majority of which involved fentanyl or fentanyl analogs. Vaccines have been proposed as a therapeutic and prophylactic strategy to offer selective and long-lasting protection against accidental or deliberate exposure to fentanyl and closely related analogs. To support the development of a clinically viable anti-opioid vaccine suitable for human use, the incorporation of adjuvants will be required to elicit high titers of high-affinity circulating antibodies specific to the target opioid. Here we demonstrate that the addition of a synthetic TLR7/8 agonist, INI-4001, but not a synthetic TLR4 agonist, INI-2002, to a candidate conjugate vaccine consisting of a fentanyl-based hapten, F
1 , conjugated to the diphtheria cross-reactive material (CRM), significantly increased generation of high-affinity F1 -specific antibody concentrations, and reduced drug distribution to the brain after fentanyl administration in mice., (© 2023. The Author(s).)- Published
- 2023
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27. IL-4 Predicts the Efficacy of a Candidate Antioxycodone Vaccine and Alters Vaccine-Specific Antibody-Secreting Cell Proliferation in Mice.
- Author
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Crouse B, Baehr C, Hicks D, and Pravetoni M
- Subjects
- Mice, Animals, Analgesics, Opioid, Germinal Center, Antibody-Producing Cells, Cell Proliferation, Interleukin-4, Vaccines
- Abstract
Opioid use disorders (OUDs) are a public health concern in the United States and worldwide. Current medications for OUDs may trigger side effects and are often heavily regulated. A novel treatment strategy to be used alone or in combination with existing medications is active immunization with antiopioid vaccines, which stimulate production of opioid-specific Abs that bind to the target drug and prevent its distribution to the brain. Although antiopioid vaccines have shown promising preclinical efficacy, prior clinical evaluations of vaccines targeting stimulants indicate that efficacy is limited to a subset of subjects who achieve optimal Ab responses. We have previously reported that depletion of IL-4 with a mAb increased opioid-specific IgG2a and total IgG, and it increased the number of germinal centers and germinal center T follicular helper cells in response to antiopioid vaccines via type I IL-4 signaling. The current study further investigates the mechanisms associated with IL-4-mediated increases in efficacy and whether IL-4 depletion affects specific processes involved in germinal center formation, including affinity maturation, class switching, and plasma cell differentiation in mice. Additionally, results demonstrate that preimmunization production of IL-4 after ex vivo whole blood stimulation predicted in vivo vaccine-induced Ab titers in outbred mice. Such mechanistic studies are critical for rational design of next-generation vaccine formulations, and they support the use of IL-4 as a predictive biomarker in ongoing OUD vaccine clinical studies., (Copyright © 2023 by The American Association of Immunologists, Inc.)
- Published
- 2023
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28. Anti-Strychnine Immunoconjugate Reduces the Effects of Strychnine-Induced Toxicity in Mice.
- Author
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Baehr C, Kassick AJ, Vigliaturo J, Luengas D, Khaimraj A, Pravetoni M, Averick SE, and Raleigh MD
- Subjects
- Mice, Animals, Anticonvulsants pharmacology, Seizures chemically induced, Seizures drug therapy, Seizures prevention & control, Brain, Strychnine adverse effects, Immunoconjugates adverse effects
- Abstract
Strychnine poisoning induces seizures that result in loss of control of airway muscles, leading to asphyxiation and subsequent death. Current treatment options are limited, requiring hands-on medical care and isolation to low-stimulus environments. Anticonvulsants and muscle relaxants have shown limited success in cases of severe toxicity. Furthermore, nonfatal strychnine poisoning is likely to result in long-term muscular and cognitive damage. Due to its potency, accessibility, and lack of effective antidotes, strychnine poses a unique threat for mass casualty incidents. As a first step toward developing an anti-strychnine immunotherapy to reduce or prevent strychnine-induced seizures, a strychnine vaccine was synthesized using subunit keyhole limpet hemocyanin. Mice were vaccinated with the strychnine immunoconjugate and then given a 0.75 mg/kg IP challenge of strychnine and observed for seizures for 30 min. Vaccination reduced strychnine-induced events, and serum strychnine levels were increased while brain strychnine levels were decreased in vaccinated animals compared to the control. These data demonstrate that strychnine-specific antibodies can block the seizure-inducing effects of strychnine and could be used to develop a therapeutic for strychnine poisoning.
- Published
- 2023
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29. The potential role of opioid vaccines and monoclonal antibodies in the opioid overdose crisis.
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Martinez S, Harris H, Chao T, Luba R, Pravetoni M, Comer SD, and Jones JD
- Subjects
- Humans, Analgesics, Opioid therapeutic use, Antibodies, Monoclonal adverse effects, Opiate Overdose, Drug Overdose prevention & control, Opioid-Related Disorders
- Published
- 2023
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30. A trypanosome-derived immunotherapeutics platform elicits potent high-affinity antibodies, negating the effects of the synthetic opioid fentanyl.
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Triller G, Vlachou EP, Hashemi H, van Straaten M, Zeelen JP, Kelemen Y, Baehr C, Marker CL, Ruf S, Svirina A, Chandra M, Urban K, Gkeka A, Kruse S, Baumann A, Miller AK, Bartel M, Pravetoni M, Stebbins CE, Papavasiliou FN, and Verdi JP
- Subjects
- Animals, Mice, Analgesics, Opioid, Fentanyl pharmacology, Fentanyl therapeutic use, Variant Surface Glycoproteins, Trypanosoma, Immunotherapy, Trypanosoma brucei brucei genetics, Trypanosomiasis, African drug therapy, Trypanosoma
- Abstract
Poorly immunogenic small molecules pose challenges for the production of clinically efficacious vaccines and antibodies. To address this, we generate an immunization platform derived from the immunogenic surface coat of the African trypanosome. Through sortase-based conjugation of the target molecules to the variant surface glycoprotein (VSG) of the trypanosome surface coat, we develop VSG-immunogen array by sortase tagging (VAST). VAST elicits antigen-specific memory B cells and antibodies in a murine model after deploying the poorly immunogenic molecule fentanyl as a proof of concept. We also develop a single-cell RNA sequencing (RNA-seq)-based computational method that synergizes with VAST to specifically identify memory B cell-encoded antibodies. All computationally selected antibodies bind to fentanyl with picomolar affinity. Moreover, these antibodies protect mice from fentanyl effects after passive immunization, demonstrating the ability of these two coupled technologies to elicit therapeutic antibodies to challenging immunogens., Competing Interests: Declaration of interests G.T., C.E.S., F.N.P., and J.P.V. report being shareholders of Panosome GmbH and Hepione Therapeutics and having patents planned, pending, or issued broadly relevant to the work. C.E.S. and F.N.P. report being the managing directors of Panosome GmbH. S.K. reports being a shareholder of Panosome GmbH and Hepione Therapeutics. S.R. and K.U. report being employees of Panosome GmbH. P.V., K.U., M.v.S., J.P.Z., Y.K., A.S., A.K.M, A.B., and H.H. report having patents planned, pending, or issued broadly relevant to the work. M.P. and C.B. have patents pending related to fentanyl haptens, conjugates, and mAbs against fentanyl-like compounds., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2023
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31. Structures of drug-specific monoclonal antibodies bound to opioids and nicotine reveal a common mode of binding.
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Rodarte JV, Baehr C, Hicks D, Liban TL, Weidle C, Rupert PB, Jahan R, Wall A, McGuire AT, Strong RK, Runyon S, Pravetoni M, and Pancera M
- Subjects
- Mice, Animals, Antibodies, Monoclonal chemistry, Oxycodone therapeutic use, Morphine therapeutic use, Analgesics, Opioid therapeutic use, Nicotine
- Abstract
Opioid-related fatal overdoses have reached epidemic proportions. Because existing treatments for opioid use disorders offer limited long-term protection, accelerating the development of newer approaches is critical. Monoclonal antibodies (mAbs) are an emerging treatment strategy that targets and sequesters selected opioids in the bloodstream, reducing drug distribution across the blood-brain barrier, thus preventing or reversing opioid toxicity. We previously identified a series of murine mAbs with high affinity and selectivity for oxycodone, morphine, fentanyl, and nicotine. To determine their binding mechanism, we used X-ray crystallography to solve the structures of mAbs bound to their respective targets, to 2.2 Å resolution or higher. Structural analysis showed a critical convergent hydrogen bonding mode that is dependent on a glutamic acid residue in the mAbs' heavy chain and a tertiary amine of the ligand. Characterizing drug-mAb complexes represents a significant step toward rational antibody engineering and future manufacturing activities to support clinical evaluation., Competing Interests: Declaration of interests M. Pravetoni, D.H., and C.B. are inventors of provisional and non-provisional patent applications covering the anti-fentanyl mAbs described herein., (Copyright © 2022 Elsevier Ltd. All rights reserved.)
- Published
- 2023
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32. Immunotherapeutic strategies for treating opioid use disorder and overdose.
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Luba R, Martinez S, Jones J, Pravetoni M, and Comer SD
- Subjects
- Humans, Analgesics, Opioid adverse effects, Immunotherapy, Opioid-Related Disorders drug therapy, Vaccines, Buprenorphine therapeutic use
- Abstract
Introduction: Development and implementation of effective treatments for opioid use disorder (OUD) and prevention of overdose are urgent public health needs. Though existing medications for OUD (MOUD) are effective, barriers to initiation and retention in treatment persist. Therefore, development of novel treatments, especially those may complement existing treatments, is needed., Areas Covered: This review provides an overview of vaccines for substance use disorders (SUD) and mechanisms underlying their function and efficacy. Next, we focus on existing preclinical and clinical trials of SUD vaccines. We focus briefly on related strategies before providing an expert opinion on prior, current, and future work on vaccines for OUD. We included published findings from preclinical and clinical trials found on PubMed and ScienceDirect as well as ongoing or initiated trials listed on ClinicalTrials.gov., Expert Opinion: The present opioid overdose and OUD crises necessitate urgent development and implementation of effective treatments, especially those that offer protection from overdose and can serve as adjuvants to existing medications. Promising preclinical trial results paired with careful efforts to develop vaccines that account for prior SUD vaccine shortcomings offer hope for current and future clinical trials of opioid vaccines. Clinical advantages of opioid vaccines appear to outnumber disadvantages, which may result in improved treatment options.
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- 2023
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33. Clinical Trial Design Challenges and Opportunities for Emerging Treatments for Opioid Use Disorder: A Review.
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Kiluk BD, Kleykamp BA, Comer SD, Griffiths RR, Huhn AS, Johnson MW, Kampman KM, Pravetoni M, Preston KL, Vandrey R, Bergeria CL, Bogenschutz MP, Brown RT, Dunn KE, Dworkin RH, Finan PH, Hendricks PS, Houtsmuller EJ, Kosten TR, Lee DC, Levin FR, McRae-Clark A, Raison CL, Rasmussen K, Turk DC, Weiss RD, and Strain EC
- Subjects
- Humans, Analgesics, Opioid adverse effects, Opiate Substitution Treatment, Quality of Life, Clinical Trials as Topic, Opioid-Related Disorders drug therapy, Buprenorphine therapeutic use
- Abstract
Importance: Novel treatments for opioid use disorder (OUD) are needed to address both the ongoing opioid epidemic and long-standing barriers to existing OUD treatments that target the endogenous μ-opioid receptor (MOR) system. The goal of this review is to highlight unique clinical trial design considerations for the study of emerging treatments for OUD that address targets beyond the MOR system. In November 2019, the Analgesic, Anesthetic, and Addiction Clinical Trial Translations, Innovations, Opportunities, and Networks (ACTTION) public-private partnership with the US Food and Drug Administration sponsored a meeting to discuss the current evidence regarding potential treatments for OUD, including cannabinoids, psychedelics, sedative-hypnotics, and immunotherapeutics, such as vaccines., Observations: Consensus recommendations are presented regarding the most critical elements of trial design for the evaluation of novel OUD treatments, such as: (1) stage of treatment that will be targeted (eg, seeking treatment, early abstinence/detoxification, long-term recovery); (2) role of treatment (adjunctive with or independent of existing OUD treatments); (3) primary outcomes informed by patient preferences that assess opioid use (including changes in patterns of use), treatment retention, and/or global functioning and quality of life; and (4) adverse events, including the potential for opioid-related relapse or overdose, especially if the patient is not simultaneously taking maintenance MOR agonist or antagonist medications., Conclusions and Relevance: Applying the recommendations provided here as well as considering input from people with lived experience in the design phase will accelerate the development, translation, and uptake of effective and safe therapeutics for individuals struggling with OUD.
- Published
- 2023
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34. Advancing humanized monoclonal antibody for counteracting fentanyl toxicity towards clinical development.
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Hicks D, Baehr C, Silva-Ortiz P, Khaimraj A, Luengas D, Hamid FA, and Pravetoni M
- Subjects
- Animals, Humans, Mice, Rats, Antibodies, Monoclonal therapeutic use, Complement System Proteins, Antibodies, Monoclonal, Humanized therapeutic use, Fentanyl immunology, Fentanyl toxicity, Narcotic Antagonists chemistry, Narcotic Antagonists immunology
- 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.
- Published
- 2022
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35. Pre-clinical safety and toxicology profile of a candidate vaccine to treat oxycodone use disorder.
- Author
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Hamid FA, Marker CL, Raleigh MD, Khaimraj A, Winston S, Pentel PR, and Pravetoni M
- Subjects
- Aluminum, Animals, Oxycodone therapeutic use, Rats, Rats, Sprague-Dawley, Vaccines, Conjugate, Drug Overdose, Opioid-Related Disorders drug therapy, Opioid-Related Disorders prevention & control
- Abstract
Opioid use disorders (OUD) and overdose represent a public health threat, resulting in thousands of deaths annually worldwide. Vaccines offer a promising treatment for OUD and potentially the prevention of fatal overdoses. The Oxy(Gly)
4 -sKLH Conjugate Vaccine, Adsorbed (Oxy(Gly)4 -sKLH) has shown promising pre-clinical efficacy at reducing the behavioral and pharmacological effects of oxycodone. To support its clinical evaluation, a GLP toxicology study was performed to address the safety of Oxy(Gly)4 -sKLH. Sprague Dawley rats were vaccinated with either aluminum adjuvant (alum) or vaccine adsorbed on alum. Low and high doses of Oxy(Gly)4 -sKLH, equivalent to a 1X or 47X human dose, respectively, were administered every two weeks for a total of four vaccinations. Both vaccine doses induced high antibody titers. Vaccine-related toxicity was assessed postmortem in one experimental group after receiving the fourth immunization of the vaccine's high dose. For the remaining experimental groups, rats were challenged with 1.5 mg/kg/day s.c. oxycodone for 7 days after the fourth vaccination to assess whether concurrent exposure to oxycodone in vaccinated animals resulted in toxicity. All rats, except a subset of the aluminum control and the high dose vaccine groups, were sacrificed following oxycodone exposure. These subsets were allowed a four weeks recovery period prior to euthanasia. In this study, no Oxy(Gly)4 -sKLH-related hematology, clinical chemistry, urinalysis, body weight, organ weight, or anatomic pathology toxicological findings were observed. These results demonstrate that the Oxy(Gly)4 -sKLH vaccine is well tolerated, is immunogenic even at low doses, and does not produce undesired side effects in rats., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Scott Winston reports a relationship with Winston Biopharmaceutical Consulting that includes: consulting or advising., (Copyright © 2022 Elsevier Ltd. All rights reserved.)- Published
- 2022
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36. Preclinical Efficacy and Selectivity of Vaccines Targeting Fentanyl, Alfentanil, Sufentanil, and Acetylfentanyl in Rats.
- Author
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Baehr C, Robinson C, Kassick A, Jahan R, Gradinati V, Averick SE, Runyon SP, and Pravetoni M
- Abstract
The ongoing public health emergency of opioid use disorders (OUD) and overdose in the United States is largely driven by fentanyl and its related analogues and has resulted in over 75 673 deaths in 2021. Immunotherapeutics such as vaccines have been investigated as a potential interventional strategy complementary to current pharmacotherapies to reduce the incidence of OUD and opioid-related overdose. Given the importance of targeting structurally distinct fentanyl analogues, this study compared a previously established lead conjugate vaccine (F
1 -CRM) to a series of novel vaccines incorporating haptens derived from alfentanil and acetylfentanyl (F8, 9a, 9b, 10 ), and evaluated their efficacy against drug-induced pharmacological effects in rats. While no vaccine tested provided significant protection against alfentanil, lead formulations were effective in reducing antinociception, respiratory depression, and bradycardia elicited by fentanyl, sufentanil, and acetylfentanyl. Compared with control, vaccination with F1 -CRM also reduced drug levels in the brain of rats challenged with lethal doses of fentanyl. These data further support investigation of F1 -CRM as a candidate vaccine against fentanyl and selected analogues., Competing Interests: The authors declare the following competing financial interest(s): Pravetoni, Averick, and Runyon are inventors of provisional application No. 62/989,41, "Fentanyl haptens, fentanyl hapten conjugates, and methods for making and using." The other authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)- Published
- 2022
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37. Pharmacological Profiling of Antifentanyl Monoclonal Antibodies in Combination with Naloxone in Pre- and Postexposure Models of Fentanyl Toxicity.
- Author
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Baehr CA, Wu MM, Pandit SG, Arias-Umana J, AuCoin D, and Pravetoni M
- Subjects
- Analgesics, Opioid therapeutic use, Animals, Antibodies, Monoclonal pharmacology, Antibodies, Monoclonal therapeutic use, Fentanyl, Humans, Naloxone pharmacology, Naloxone therapeutic use, Narcotic Antagonists pharmacology, Pandemics, Rats, COVID-19, Drug Overdose drug therapy, Respiratory Insufficiency drug therapy
- Abstract
The incidence of fatal drug overdoses in the United States is an alarming public health threat that has been exacerbated by the COVID-19 pandemic, resulting in over 100,000 deaths between April 2020 and April 2021. A significant portion of this is attributable to widespread access to fentanyl and other synthetic opioids, alone or in combination with heroin or psychostimulants, such as cocaine or methamphetamine. Monoclonal antibodies (mAb) offer prophylactic and therapeutic interventions against opioid overdose by binding opioids in serum, reducing distribution of drug to the brain and other organs. Here, we investigated the efficacy of a leading antifentanyl mAb, clone HY6-F9, in reversal and prevention of fentanyl-induced toxicity compared with the opioid receptor antagonist naloxone (NLX) in rats. In postexposure models, rats were challenged with fentanyl, followed by HY6-F9, NLX, or both. HY6-F9 reversed fentanyl-induced antinociception, respiratory depression, and bradycardia, and rats retained protection against additional challenges for at least 1 week. Although intravenous NLX reversed fentanyl-induced respiratory depression more rapidly than mAb alone, kinetics of reversal by intravenous mAb were similar to subcutaneous NLX. Coadministration of mAb and NLX provided greater protection than individual treatments against high doses of fentanyl. Prophylactic administration of mAb reduced the ED
50 of NLX approximately twofold against 2.25 mg/kg of fentanyl. Finally, mAb sequestered fentanyl and its metabolite norfentanyl in serum and reduced brain concentrations of fentanyl. These results support the translation of mAb as medical interventions alone or in combination with NLX to prevent and reverse fentanyl-related overdose. SIGNIFICANCE STATEMENT: Fentanyl-related overdoses have increased dramatically in the US and worldwide. Currently, approved pharmacotherapies for treatment of opioid use disorder and reversal of overdose are not sufficient to curb the incidence of opioid-related deaths. Additionally, fentanyl and its potent analogs present a potential risk from use in deliberate poisoning or chemical attacks. This study demonstrates the use of monoclonal antibodies as a countermeasure to fentanyl-induced toxicity in pre- and postexposure scenarios, supporting their use in combination with the opioid antagonist naloxone., (Copyright © 2022 by The American Society for Pharmacology and Experimental Therapeutics.)- Published
- 2022
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38. Efficacy and Selectivity of Monovalent and Bivalent Vaccination Strategies to Protect against Exposure to Carfentanil, Fentanyl, and Their Mixtures in Rats.
- Author
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Crouse B, Wu MM, Gradinati V, Kassick AJ, Song D, Jahan R, Averick S, Runyon S, Comer SD, and Pravetoni M
- Abstract
Drug-related fatal overdoses have significantly increased in the past decade due to the widespread availability of illicit fentanyl and other potent synthetic opioids such as carfentanil. Deliberate or accidental consumption or exposure to carfentanil, fentanyl, and their mixture induces respiratory depression and bradycardia that can be difficult to reverse with the opioid receptor antagonist naloxone. Vaccines offer a promising strategy to reduce the incidence of fatalities associated with fentanyl-related substances, as well as treatment for opioid use disorder (OUD). This study reports monovalent and bivalent vaccination strategies that elicit polyclonal antibody responses effective in protecting against the pharmacological actions of carfentanil, fentanyl, or carfentanil/fentanyl mixtures. Rats were prophylactically immunized with individual conjugate vaccines containing either carfentanil- or fentanyl-based haptens, or their combination in bivalent vaccine formulations, and then challenged with carfentanil, fentanyl, or their mixture. First, these studies identified a lead vaccine protective against carfentanil-induced antinociception, respiratory depression, and bradycardia. Then, efficacy against both carfentanil and fentanyl was achieved through bivalent vaccination strategies that combined lead anti-carfentanil and anti-fentanyl vaccines via either heterologous prime/boost or co-administration immunization regimens. These preclinical data support the development of vaccines as a viable strategy to prevent toxicity from exposure to excessive doses of carfentanil, fentanyl, or their mixtures., Competing Interests: The authors declare the following competing financial interest(s): Pravetoni is the inventor of Fentanyl hapten-conjugates and methods for making and using same, Application No. 62/932,757. Pravetoni, Averick, and Runyon are inventors of Fentanyl haptens, fentanyl hapten conjugates, and methods for making and using the same, Application No. 62/989,417. The other authors declare no conflict of interest., (© 2022 American Chemical Society.)
- Published
- 2022
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39. A Membrane-Modulated Centrifugal Microdevice for Enzyme-Linked Immunosorbent Assay-Based Detection of Illicit and Misused Drugs.
- Author
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Dignan LM, Woolf MS, Ross JA, Baehr C, Holstege CP, Pravetoni M, and Landers JP
- Subjects
- Colorimetry, Enzyme-Linked Immunosorbent Assay, Humans, Microfluidics, Pharmaceutical Preparations, Substance Abuse Detection
- Abstract
Increased opioid use and misuse have imposed large analytical demands across clinical and forensic sectors. Due to the absence of affordable, accurate, and simple on-site tests (e.g., point of interdiction and bedside), analysis is primarily conducted in centralized laboratories via time-consuming, labor-intensive methods. Many healthcare facilities do not have such analytical capabilities and must send samples to commercial laboratories, increasing turnaround time and care costs, as well as delaying public health warnings regarding the emergence of specific substances. Enzyme-linked immunosorbent assays (ELISAs) are used ubiquitously, despite lengthy workflows that require substantial manual intervention. Faster, reliable analytics are desperately needed to mitigate the mortality and morbidity associated with the current substance use epidemic. We describe one such alternative─a portable centrifugal microfluidic ELISA system that supplants repetitive pipetting with rotationally controlled fluidics. Embedded cellulosic membranes act as microvalves, permitting flow only when centrifugally generated hydraulic pressure exceeds their liquid entry pressure. These features enable stepwise reagent introduction, incubation, and removal simply by tuning rotational frequency. We demonstrate the success of this platform through sensitive, specific colorimetric detection of opiates, a subclass of opioids naturally derived from the opium poppy. Objective image analysis eliminated subjectivity in human color perception and permitted reliable detection of opiates in buffer and artificial urine at the ng/μL range. Opiates were clearly differentiated from other drug classes without interference from common adulterants known to cause false positive results in current colorimetric field tests. Eight samples were simultaneously analyzed in under 1 h, a marked reduction from the traditional multiday timeline. This approach could permit rapid, automatable ELISA-based drug detection outside of traditional laboratories by nontechnical personnel.
- Published
- 2021
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40. Housing conditions and microbial environment do not affect the efficacy of vaccines for treatment of opioid use disorders in mice and rats.
- Author
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Crouse B, Zhang L, Robinson C, Ban Y, Vigliaturo JR, Roy S, and Pravetoni M
- Subjects
- Animals, Housing Quality, Humans, Male, Mice, RNA, Ribosomal, 16S, Rats, Vaccine Efficacy, Opioid-Related Disorders, Vaccines
- Abstract
Vaccines offer a promising prophylactic and therapeutic intervention to counteract opioid use disorders (OUD) and fatal overdoses. Vaccines generate opioid-specific antibodies that bind the target opioid, reducing drug distribution to the brain and preventing drug-induced behavioral and pharmacological effects. Due to their selectivity, anti-opioid vaccines can be administered in combination with FDA-approved medications. Because patients with OUD or other substance use disorders may be affected by other multifactorial co-morbidities, such as infection or depression, it is important to test whether vaccine efficacy is modified by factors that may impact individual innate or adaptive immunity. To that end, this study tested whether housing conditions would affect the efficacy of two lead vaccine formulations targeting oxycodone and fentanyl in male mice and rats, and further analyzed whether differences in the gastrointestinal (GI) microbiome would be correlated with either vaccine efficacy or housing conditions. Results showed that housing mice and rats in either conventional (non-controlled) or specific pathogen-free (SPF, sterile barrier maintained) environment did not affect vaccine-induced antibody responses against oxycodone and fentanyl, nor their efficacy against oxycodone- and fentanyl-induced antinociception, respiratory depression, and bradycardia. Differences in the GI microbiome detected via 16S rRNA gene sequencing were related to the housing environment. This study supports use of anti-opioid vaccines in clinical populations that may display deficits in microbiome function.
- Published
- 2021
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41. Covalently Loaded Naloxone Nanoparticles as a Long-Acting Medical Countermeasure to Opioid Poisoning.
- Author
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Kassick AJ, Wu M, Luengas D, Ebqa'ai M, Tharika Nirmani LP, Tomycz N, Nelson TL, Pravetoni M, Raleigh MD, and Averick S
- Abstract
The mu opioid receptor antagonist naloxone has been a vital, long-standing countermeasure in the ongoing battle against opioid use disorders (OUD) and toxicity. However, due to its distinctive short elimination half-life, naloxone has shown diminished efficacy in cases of synthetic opioid poisoning as larger or repeated doses of the antidote have been required to achieve adequate reversal of severe respiratory depression and prevent episodes of renarcotization. This report describes the synthesis, characterization, and in vivo evaluation of a novel, nanoparticle-based naloxone formulation that provides extended protection against the toxic effects of the powerful synthetic opioid fentanyl. The strategy was predicated on a modified two-step protocol involving the synthesis and subsequent nanoprecipitation of a poly(lactic- co -glycolic acid) polymer scaffold bearing a covalently linked naloxone chain end (drug loading ∼7% w/w). Pharmacokinetic evaluation of the resulting covalently loaded naloxone nanoparticles ( c NLX-NP) revealed an elimination half-life that was 34 times longer than high dose free naloxone (10 mg/kg) in male Sprague-Dawley rats. This enhancement was further demonstrated by c NLX-NP in subsequent in vivo studies affording protection against fentanyl-induced respiratory depression and antinociception for up to 48 h following a single intramuscular injection. These discoveries support further investigation of c NLX-NP as a potential therapeutic to reverse overdose and prevent renarcotization from fentanyl and its potent analogs., Competing Interests: The authors declare no competing financial interest., (© 2021 American Chemical Society.)
- Published
- 2021
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42. Contribution of Antibody-Mediated Effector Functions to the Mechanism of Efficacy of Vaccines for Opioid Use Disorders.
- Author
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Huseby Kelcher AM, Baehr CA, Hamid FA, Hart GT, and Pravetoni M
- Subjects
- Animals, Antibodies, Monoclonal, Mice, Oxycodone, Opioid-Related Disorders, Vaccines
- Abstract
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')2 s 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., (Copyright © 2021 by The American Association of Immunologists, Inc.)- Published
- 2021
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43. Fentanyl conjugate vaccine by injected or mucosal delivery with dmLT or LTA1 adjuvants implicates IgA in protection from drug challenge.
- Author
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Stone AE, Scheuermann SE, Haile CN, Cuny GD, Velasquez ML, Linhuber JP, Duddupudi AL, Vigliaturo JR, Pravetoni M, Kosten TA, Kosten TR, and Norton EB
- Abstract
Fentanyl is a major contributor to the devastating increase in overdose deaths from substance use disorders (SUD). A vaccine targeting fentanyl could be a powerful immunotherapeutic. Here, we evaluated adjuvant and delivery strategies for conjugate antigen vaccination with fentanyl-based haptens. We tested adjuvants derived from the heat-labile toxin of E. coli including dmLT and LTA1 by intramuscular, sublingual or intranasal delivery. Our results show anti-fentanyl serum antibodies and antibody secreting cells in the bone-marrow after vaccination with highest levels observed with an adjuvant (alum, dmLT, or LTA1). Vaccine adjuvanted with LTA1 or dmLT elicited the highest levels of anti-fentanyl antibodies, whereas alum achieved highest levels against the carrier protein. Vaccination with sublingual dmLT or intranasal LTA1 provided the most robust blockade of fentanyl-induced analgesia and CNS penetration correlating strongly to anti-FEN IgA. In conclusion, this study demonstrates dmLT or LTA1 adjuvant as well as mucosal delivery may be attractive strategies for improving the efficacy of vaccines against SUD.
- Published
- 2021
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44. Countermeasures for Preventing and Treating Opioid Overdose.
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France CP, Ahern GP, Averick S, Disney A, Enright HA, Esmaeli-Azad B, Federico A, Gerak LR, Husbands SM, Kolber B, Lau EY, Lao V, Maguire DR, Malfatti MA, Martinez G, Mayer BP, Pravetoni M, Sahibzada N, Skolnick P, Snyder EY, Tomycz N, Valdez CA, and Zapf J
- Subjects
- Animals, Congresses as Topic, Drug Overdose etiology, Drug Overdose mortality, Humans, Naloxone adverse effects, Narcotic Antagonists adverse effects, Opioid-Related Disorders complications, Opioid-Related Disorders mortality, Prognosis, Risk Assessment, Risk Factors, Analgesics, Opioid adverse effects, Drug Overdose therapy, Medical Countermeasures, Naloxone therapeutic use, Narcotic Antagonists therapeutic use, Opioid Epidemic mortality, Opioid-Related Disorders therapy
- Abstract
The only medication available currently to prevent and treat opioid overdose (naloxone) was approved by the US Food and Drug Administration (FDA) nearly 50 years ago. Because of its pharmacokinetic and pharmacodynamic properties, naloxone has limited utility under some conditions and would not be effective to counteract mass casualties involving large-scale deployment of weaponized synthetic opioids. To address shortcomings of current medical countermeasures for opioid toxicity, a trans-agency scientific meeting was convened by the US National Institute of Allergy and Infectious Diseases/National Institutes of Health (NIAID/NIH) on August 6 and 7, 2019, to explore emerging alternative approaches for treating opioid overdose in the event of weaponization of synthetic opioids. The meeting was initiated by the Chemical Countermeasures Research Program (CCRP), was organized by NIAID, and was a collaboration with the National Institute on Drug Abuse/NIH (NIDA/NIH), the FDA, the Defense Threat Reduction Agency (DTRA), and the Biomedical Advanced Research and Development Authority (BARDA). This paper provides an overview of several presentations at that meeting that discussed emerging new approaches for treating opioid overdose, including the following: (1) intranasal nalmefene, a competitive, reversible opioid receptor antagonist with a longer duration of action than naloxone; (2) methocinnamox, a novel opioid receptor antagonist; (3) covalent naloxone nanoparticles; (4) serotonin (5-HT)
1A receptor agonists; (5) fentanyl-binding cyclodextrin scaffolds; (6) detoxifying biomimetic "nanosponge" decoy receptors; and (7) antibody-based strategies. These approaches could also be applied to treat opioid use disorder., (© 2020 The Authors Clinical Pharmacology & Therapeutics © 2020 American Society for Clinical Pharmacology and Therapeutics.)- Published
- 2021
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45. Therapeutic and Prophylactic Vaccines to Counteract Fentanyl Use Disorders and Toxicity.
- Author
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Robinson C, Gradinati V, Hamid F, Baehr C, Crouse B, Averick S, Kovaliov M, Harris D, Runyon S, Baruffaldi F, LeSage M, Comer S, and Pravetoni M
- Subjects
- Animals, Bacterial Proteins chemistry, Bacterial Proteins immunology, Cattle, Diphtheria Toxin chemistry, Diphtheria Toxin immunology, Female, Haptens chemistry, Haptens immunology, Hemocyanins chemistry, Hemocyanins immunology, Male, Mice, Inbred BALB C, Piperidines chemical synthesis, Piperidines immunology, Proof of Concept Study, Rats, Sprague-Dawley, Serum Albumin, Bovine chemistry, Serum Albumin, Bovine immunology, Sufentanil immunology, Fentanyl immunology, Opioid-Related Disorders prevention & control, Opioid-Related Disorders therapy, Vaccines immunology
- Abstract
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.
- Published
- 2020
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46. Monoclonal Antibodies Counteract Opioid-Induced Behavioral and Toxic Effects in Mice and Rats.
- Author
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Baehr C, Kelcher AH, Khaimraj A, Reed DE, Pandit SG, AuCoin D, Averick S, and Pravetoni M
- Subjects
- Animals, Female, Immunization, Male, Mice, Rats, Analgesics, Opioid immunology, Analgesics, Opioid toxicity, Antibodies, Monoclonal immunology, Behavior, Animal
- Abstract
Monoclonal antibodies (mAbs) and vaccines have been proposed as medical countermeasures to treat opioid use disorder (OUD) and prevent opioid overdose. In contrast to current pharmacotherapies (e.g., methadone, buprenorphine, naltrexone, and naloxone) for OUD and overdose, which target brain opioid receptors, mAbs and vaccine-generated polyclonal antibodies sequester the target opioid in the serum and reduce drug distribution to the brain. Furthermore, mAbs offer several potential clinical benefits over approved medications, such as longer serum half-life, higher selectivity, reduced side effects, and no abuse liability. Using magnetic enrichment to isolate opioid-specific B cell lymphocytes prior to fusion with myeloma partners, this study identified a series of murine hybridoma cell lines expressing mAbs with high affinity for opioids of clinical interest, including oxycodone, heroin and its active metabolites, and fentanyl. In mice, passive immunization with lead mAbs against oxycodone, heroin, and fentanyl reduced drug-induced antinociception and the distribution of the target opioid to the brain. In mice and rats, mAb pretreatment reduced fentanyl-induced respiratory depression and bradycardia, two risk factors for opioid-related overdose fatality. Overall, these results support use of mAbs to counteract toxic effects of opioids and other chemical threats. SIGNIFICANCE STATEMENT: The incidence of fatal overdoses due to the widespread access to heroin, prescription opioids, and fentanyl suggests that current Food and Drug Administration-approved countermeasures are not sufficient to mitigate the opioid epidemic. Monoclonal antibodies (mAbs) may provide acute protection from overdose by binding to circulating opioids in serum. Use of mAbs prophylactically, or after exposure in combination with naloxone, may reduce hospitalization and increase survival., (Copyright © 2020 by The American Society for Pharmacology and Experimental Therapeutics.)
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- 2020
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47. Sex differences in oral oxycodone self-administration and stress-primed reinstatement in rats.
- Author
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Fulenwider HD, Nennig SE, Hafeez H, Price ME, Baruffaldi F, Pravetoni M, Cheng K, Rice KC, Manvich DF, and Schank JR
- Subjects
- Analgesics, Opioid blood, Animals, Drug-Seeking Behavior drug effects, Estrous Cycle drug effects, Extinction, Psychological drug effects, Female, Male, Naloxone administration & dosage, Neurokinin-1 Receptor Antagonists pharmacology, Oxycodone blood, Rats, Rats, Wistar, Receptors, Opioid, mu antagonists & inhibitors, Analgesics, Opioid administration & dosage, Oxycodone administration & dosage, Prescription Drug Misuse, Receptors, Neurokinin-1 physiology, Self Administration
- Abstract
The opioid epidemic has become a severe public health problem, with approximately 130 opioid-induced deaths occurring each day in the United States. Prescription opioids are responsible for approximately 40% of these deaths. Oxycodone is one of the most commonly abused prescription opioids, but despite its prevalent misuse, the number of preclinical studies investigating oxycodone-seeking behaviors is relatively limited. Furthermore, preclinical oxycodone studies that include female subjects are even more scarce, and it is critical that future work includes both sexes. Additionally, the oral route of administration is one of the most common routes for recreational users, especially in the early stages of drug experimentation. However, currently, only two studies have been published investigating operant oral oxycodone self-administration in rodents. Therefore, the primary goal of the present study was to establish an oral oxycodone operant self-administration model in adult male and female rats, as well as to examine a potential mechanism of stress-primed reinstatement. We found that females consumed significantly more oral oxycodone than males in operant self-administration sessions. We also found that active oxycodone self-administration was reduced by mu opioid receptor antagonism and by substitution of water for oxycodone solution. Lastly, we induced stress-primed reinstatement and found that this behavior was significantly attenuated by antagonism of the neurokinin-1 receptor, consistent with our prior work examining stress-induced reinstatement of alcohol- and cocaine-seeking., (© 2019 Society for the Study of Addiction.)
- Published
- 2020
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48. Mechanisms of interleukin 4 mediated increase in efficacy of vaccines against opioid use disorders.
- Author
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Crouse B, Robinson C, Huseby Kelcher A, Laudenbach M, Abrahante JE, and Pravetoni M
- Abstract
Opioid use disorders (OUD) affect over 27 million people worldwide. Anti-opioid vaccines offer a promising strategy to treat OUD and prevent overdose. Using immunomodulation of cytokine signaling to increase vaccine efficacy, this study found that blocking IL-4 improved the efficacy of vaccines targeting oxycodone and fentanyl in male and female mice. Genetic deletion of the IL-4 receptor, STAT6, or antibody-based depletion of IL-13, did not increase vaccine efficacy against opioids, suggesting the involvement of type I IL-4 receptors. Enhancement of vaccine efficacy with blockade of IL-4 was associated with improved germinal center formation in secondary lymphoid organs and selective transcriptome signatures in the activated CD4
+ T cell population subset. These data suggest that IL-4 is both a pharmacological target and a potential biomarker of vaccine efficacy against OUD., Competing Interests: Competing interestsM.P. is the inventor of “Cytokine Signaling Immunomodulators and Methods”, and “fentanyl hapten-conjugates and methods for making and using same” patents. The anti-IL-13 mAb was obtained from Genentech under a standard Material Transfer Agreement. All authors declare that there are no other competing interests., (© The Author(s) 2020.)- Published
- 2020
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49. Interruption of continuous opioid exposure exacerbates drug-evoked adaptations in the mesolimbic dopamine system.
- Author
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Lefevre EM, Pisansky MT, Toddes C, Baruffaldi F, Pravetoni M, Tian L, Kono TJY, and Rothwell PE
- Subjects
- Animals, Brain, Dopamine, Female, Male, Mice, Morphine, Nucleus Accumbens, Analgesics, Opioid, Pharmaceutical Preparations
- Abstract
Drug-evoked adaptations in the mesolimbic dopamine system are postulated to drive opioid abuse and addiction. These adaptations vary in magnitude and direction following different patterns of opioid exposure, but few studies have systematically manipulated the pattern of opioid administration while measuring neurobiological and behavioral impact. We exposed male and female mice to morphine for one week, with administration patterns that were either intermittent (daily injections) or continuous (osmotic minipump infusion). We then interrupted continuous morphine exposure with either naloxone-precipitated or spontaneous withdrawal. Continuous morphine exposure caused tolerance to the psychomotor-activating effects of morphine, whereas both intermittent and interrupted morphine exposure caused long-lasting psychomotor sensitization. Given links between locomotor sensitization and mesolimbic dopamine signaling, we used fiber photometry and a genetically encoded dopamine sensor to conduct longitudinal measurements of dopamine dynamics in the nucleus accumbens. Locomotor sensitization caused by interrupted morphine exposure was accompanied by enhanced dopamine signaling in the nucleus accumbens. To further assess downstream consequences on striatal gene expression, we used next-generation RNA sequencing to perform genome-wide transcriptional profiling in the nucleus accumbens and dorsal striatum. The interruption of continuous morphine exposure exacerbated drug-evoked transcriptional changes in both nucleus accumbens and dorsal striatum, dramatically increasing differential gene expression and engaging unique signaling pathways. Our study indicates that opioid-evoked adaptations in brain function and behavior are critically dependent on the pattern of drug administration, and exacerbated by interruption of continuous exposure. Maintaining continuity of chronic opioid administration may, therefore, represent a strategy to minimize iatrogenic effects on brain reward circuits.
- Published
- 2020
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50. Combining a Candidate Vaccine for Opioid Use Disorders with Extended-Release Naltrexone Increases Protection against Oxycodone-Induced Behavioral Effects and Toxicity.
- Author
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Raleigh MD, Accetturo C, and Pravetoni M
- Subjects
- Analgesics, Opioid pharmacology, Animals, Brain drug effects, Brain metabolism, Hemocyanins pharmacology, Male, Narcotic Antagonists pharmacology, Rats, Delayed-Action Preparations pharmacology, Naltrexone pharmacology, Opioid-Related Disorders drug therapy, Oxycodone pharmacology, Vaccines administration & dosage
- Abstract
Opioid use disorders (OUDs) and opioid-related fatal overdoses are a significant public health concern in the United States and worldwide. To offer more effective medical interventions to treat or prevent OUD, antiopioid vaccines are in development that reduce the distribution of the targeted opioids to brain and subsequently reduce the associated behavioral and toxic effects. It is of critical importance that antiopioid vaccines do not interfere with medications that treat OUD. Hence, this study tested the preclinical proof of concept of combining a candidate oxycodone vaccine [oxycodone-keyhole limpet hemocyanin (OXY-KLH)] with an FDA-approved extended-release naltrexone (XR-NTX) depot formulation in rats. The effects of XR-NTX on oxycodone-induced motor activity and antinociception were first assessed in nonvaccinated naïve rats to establish a baseline for subsequent studies. Next, OXY-KLH and XR-NTX were coadministered to determine whether the combination would affect the efficacy of each individual treatment, and it was found that the combination of OXY-KLH and XR-NTX offered greater efficacy in reducing oxycodone-induced motor activity, thigmotaxis, antinociception, and respiratory depression over a range of repeated or escalating oxycodone doses in rats. These data support the feasibility of combining antibody-based therapies with opioid receptor antagonists to provide greater or prolonged protection against opioid-related toxicity or overdose. Combining antiopioid vaccines with XR-NTX may provide prophylactic measures to subjects at risk of relapse and accidental or deliberate exposure. Combination therapy may extend to other biologics (e.g., monoclonal antibodies) and medications against substance use disorders. SIGNIFICANCE STATEMENT: Opioid use disorders (OUDs) remain a major problem worldwide, and new therapies are needed. This study reports on the combination of an oxycodone vaccine [oxycodone-keyhole limpet hemocyanin (OXY-KLH)] with a currently approved OUD therapy, extended-release naltrexone (XR-NTX). Results demonstrated that XR-NTX did not interfere with OXY-KLH efficacy, and combination of low doses of XR-NTX with vaccine was more effective than each individual treatment alone to reduce behavioral and toxic effects of oxycodone, suggesting that combining OXY-KLH with XR-NTX may improve OUD outcomes., (Copyright © 2020 by The American Society for Pharmacology and Experimental Therapeutics.)
- Published
- 2020
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