Abstract
Rationale
Alcohol use disorder (AUD) is a debilitating physiological and psychiatric disorder which affects individuals globally. The current pharmacological interventions to treat AUD are limited, and hence there is an urgent need for a novel pharmacological therapy which can be effective and safe across the population.
Objective
We aimed to investigate a novel neutral cannabinoid receptor-1 (CB1R) antagonist, AM6527, in several preclinical models of ethanol consumption using male and female C57BL6/J mice.
Methods
Independent groups of male and female mice were subjected to repeated cycles of drinking in the dark (DID), or intermittent access to alcohol (IAA) procedures. Twenty minutes prior to ethanol access in each procedure, animals were treated with intraperitoneal injections of either 1, 3, and 10 mg/kg of AM6527 or its respective vehicle. Acamprosate (100, 200, 300, and 400 mg/kg) or its respective vehicle was used as a positive control. Separate groups of male mice were subjected to a chain schedule of ethanol reinforcement to gain access to ethanol wherein completion of a fixed interval (FI; 5 min) schedule (link 1: “Seeking”) was reinforced with continuous access to ethanol (fixed ratio; FR1) for up to 1.8 g/kg (link 2: “consumption”). All the animals were treated with 1, 3, and 10 mg/kg of AM6527 or its respective vehicle 20 mins prior to the start of the FI chain of the procedure. Separately, AM6527 was also evaluated in male and female mice undergoing acute ethanol withdrawal following 8 weeks of intermittent or continuous access to 20% ethanol drinking.
Results
In both DID and IAA procedures, AM6527 reduced ethanol consumption in a dose-related manner in both male and female mice. AM6527 produced no tolerance in the DID procedure; mice treated with 3 mg/kg of AM6527 for 3 weeks continuously drank significantly smaller amounts of ethanol as compared to vehicle-treated mice over a period of three DID cycles. Moreover, in the IAA procedure, AM6527 caused an increase in water intake over the 24-h period. Acamprosate transiently reduced ethanol intake in male mice in both the DID and the IAA procedures but failed to produce any significant effect in female mice. AM6527 also produced a decrease in the FI responding (“ethanol seeking”) in animals trained to self-administer ethanol. Lastly, AM6527 mitigated neurological withdrawal signs, i.e., handling induced convulsions (HIC) in mice undergoing acute ethanol withdrawal.
Conclusions
Current findings support previous studies with CB1R neutral antagonist in reducing voluntary ethanol intake and seeking behavior. Based on results shown in this work, AM6527 can be developed as a first in class CB1R neutral antagonist to treat AUD in both males and females.
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References
Arnone M, Maruani J, Chaperon F, Thiebot MH, Poncelet M, Soubrie P, Le Fur G (1997) Selective inhibition of sucrose and ethanol intake by SR 141716, an antagonist of central cannabinoid (CB1) receptors. Psychopharmacology 132(1):104–106. https://doi.org/10.1007/s002130050326
Balla A, Dong B, Shilpa BM, Vemuri K, Makriyannis A, Pandey SC, Sershen H, Suckow RF, Vinod KY (2018) Cannabinoid-1 receptor neutral antagonist reduces binge-like alcohol consumption and alcohol-induced accumbal dopaminergic signaling. Neuropharmacology 131:200–208. https://doi.org/10.1016/j.neuropharm.2017.10.040
Basavarajappa BS, Joshi V, Shivakumar M, Subbanna S (2019) Distinct functions of endogenous cannabinoid system in alcohol abuse disorders. Br J Pharmacol 176(17):3085–3109. https://doi.org/10.1111/bph.14780
Christensen R, Kristensen PK, Bartels EM, Bliddal H, Astrup A (2007) Efficacy and safety of the weight-loss drug rimonabant: a meta-analysis of randomised trials. Lancet 370(9600):1706–1713. https://doi.org/10.1016/s0140-6736(07)61721-8
Colombo G, Agabio R, Fà M, Guano L, Lobina C, Loche A, Reali R, Gessa GL (1998) Reduction of voluntary ethanol intake in ethanol-preferring sP rats by the cannabinoid antagonist SR-141716. Alcohol Alcohol 33(2):126–130. https://doi.org/10.1093/oxfordjournals.alcalc.a008368
Colombo G, Serra S, Brunetti G, Gomez R, Melis S, Vacca G, Carai MM, Gessa L (2002) Stimulation of voluntary ethanol intake by cannabinoid receptor agonists in ethanol-preferring sP rats. Psychopharmacology 159(2):181–187. https://doi.org/10.1007/s002130100887
Cota D, Marsicano G, Lutz B, Vicennati V, Stalla GK, Pasquali R, Pagotto U (2003) Endogenous cannabinoid system as a modulator of food intake. Int J Obes Relat Metab Disord 27(3):289–301. https://doi.org/10.1038/sj.ijo.0802250
Covington HE 3rd, Newman EL, Tran S, Walton L, Hayek W, Leonard MZ, DeBold JF, Miczek KA (2018) The urge to fight: persistent escalation by alcohol and role of NMDA receptors in mice. Front Behav Neurosci 12:206. https://doi.org/10.3389/fnbeh.2018.00206
Crabbe JC, Colville AM, Kruse LC, Cameron AJ, Spence SE, Schlumbohm JP, Huang LC, Metten P (2012) Ethanol tolerance and withdrawal severity in high drinking in the dark selectively bred mice. Alcohol Clin Exp Res 36(7):1152–1161. https://doi.org/10.1111/j.1530-0277.2011.01715.x
Crabbe JC, Ozburn AR, Metten P, Barkley-Levenson A, Schlumbohm JP, Spence SE, Hack WR, Huang LC (2017) High drinking in the dark (HDID) mice are sensitive to the effects of some clinically relevant drugs to reduce binge-like drinking. Pharmacol Biochem Behav 160:55–62. https://doi.org/10.1016/j.pbb.2017.08.002
Dulman RS, Zhang H, Banerjee R, Krishnan HR, Dong B, Hungund BL, Vinod KY, Pandey SC (2021) CB1 receptor neutral antagonist treatment epigenetically increases neuropeptide Y expression and decreases alcohol drinking. Neuropharmacology 195:108623. https://doi.org/10.1016/j.neuropharm.2021.108623
Economidou D, Mattioli L, Cifani C, Perfumi M, Massi M, Cuomo V, Trabace L, Ciccocioppo R (2006) Effect of the cannabinoid CB1 receptor antagonist SR-141716A on ethanol self-administration and ethanol-seeking behaviour in rats. Psychopharmacology 183(4):394–403. https://doi.org/10.1007/s00213-005-0199-9
Gallate JE, Saharov T, Mallet PE, McGregor IS (1999) Increased motivation for beer in rats following administration of a cannabinoid CB1 receptor agonist. Eur J Pharmacol 370(3):233–240. https://doi.org/10.1016/s0014-2999(99)00170-3
Giuliano C, Goodlett CR, Economidou D, García-Pardo MP, Belin D, Robbins TW, Bullmore ET, Everitt BJ (2015) The novel μ-opioid receptor antagonist GSK1521498 decreases both alcohol seeking and drinking: evidence from a new preclinical model of alcohol seeking. Neuropsychopharmacology 40(13):2981–2992. https://doi.org/10.1038/npp.2015.152
Goldstein DB (1972) An animal model for testing effects of drugs on alcohol withdrawal reactions. J Pharmacol Exp Ther 183(1):14–22
Goldstein DB, Pal N (1971) Alcohol dependence produced in mice by inhalation of ethanol: grading the withdrawal reaction. Science 172(3980):288–290. https://doi.org/10.1126/science.172.3980.288
Hwa LS, Chu A, Levinson SA, Kayyali TM, DeBold JF, Miczek KA (2011) Persistent escalation of alcohol drinking in C57BL/6J mice with intermittent access to 20% ethanol. Alcohol Clin Exp Res 35(11):1938–1947. https://doi.org/10.1111/j.1530-0277.2011.01545.x
Hwa LS, Kalinichev M, Haddouk H, Poli S, Miczek KA (2014) Reduction of excessive alcohol drinking by a novel GABAB receptor positive allosteric modulator ADX71441 in mice. Psychopharmacology 231(2):333–343. https://doi.org/10.1007/s00213-013-3245-z
Hwa LS, Nathanson AJ, Shimamoto A, Tayeh JK, Wilens AR, Holly EN, Newman EL, DeBold JF, Miczek KA (2015) Aggression and increased glutamate in the mPFC during withdrawal from intermittent alcohol in outbred mice. Psychopharmacology 232(16):2889–2902. https://doi.org/10.1007/s00213-015-3925-y
Janero DR, Makriyannis A (2009) Cannabinoid receptor antagonists: pharmacological opportunities, clinical experience, and translational prognosis. Expert Opin Emerg Drugs 14(1):43–65. https://doi.org/10.1517/14728210902736568
Kangas BD, Zakarian AS, Vemuri K, Alapafuja SO, Jiang S, Nikas SP, Makriyannis A, Bergman J (2020) Cannabinoid antagonist drug discrimination in nonhuman primates. J Pharmacol Exp Ther 372(1):119–127. https://doi.org/10.1124/jpet.119.261818
Koob GF (2015) The dark side of emotion: the addiction perspective. Eur J Pharmacol 753:73–87. https://doi.org/10.1016/j.ejphar.2014.11.044
Korpi ER, Linden AM, Hytönen HR, Paasikoski N, Vashchinkina E, Dudek M, Herr DR, Hyytiä P (2017) Continuous delivery of naltrexone and nalmefene leads to tolerance in reducing alcohol drinking and to supersensitivity of brain opioid receptors. Addict Biol 22(4):1022–1035. https://doi.org/10.1111/adb.12393
Kunos G (2020) Interactions between alcohol and the endocannabinoid system [10.1111/acer.14306]. Alcohol Clin Exp Res 44(4):790–805. https://doi.org/10.1111/acer.14306
Litten RZ, Wilford BB, Falk DE, Ryan ML, Fertig JB (2016) Potential medications for the treatment of alcohol use disorder: an evaluation of clinical efficacy and safety. Subst Abus 37(2):286–298. https://doi.org/10.1080/08897077.2015.1133472
Mason BJ, Heyser CJ (2010) Acamprosate: a prototypic neuromodulator in the treatment of alcohol dependence. CNS Neurol Disord Drug Targets 9(1):23–32. https://doi.org/10.2174/187152710790966641
Mason BJ, Lehert P (2012) Acamprosate for alcohol dependence: a sex-specific meta-analysis based on individual patient data. Alcohol Clin Exp Res 36(3):497–508. https://doi.org/10.1111/j.1530-0277.2011.01616.x
Melendez RI (2011) Intermittent (every-other-day) drinking induces rapid escalation of ethanol intake and preference in adolescent and adult C57BL/6J mice. Alcohol Clin Exp Res 35(4):652–658. https://doi.org/10.1111/j.1530-0277.2010.01383.x
Miczek KA, Akdilek N, Ferreira VMM, Kenneally E, Leonard MZ, Covington HE (2022a) Excessive alcohol consumption after exposure to two types of chronic social stress: intermittent episodes vs. continuous exposure in C57BL/6J mice with a history of drinking. Psychopharmacology 239(10):3287–3296. https://doi.org/10.1007/s00213-022-06211-8
Miczek KA, Akdilek N, Ferreira VMM, Leonard MZ, Marinelli LR, Covington HE 3rd. (2022b) To fight or not to fight: activation of the mPFC during decision to engage in aggressive behavior after ethanol consumption in a novel murine model. Psychopharmacology 239(10):3249–3261. https://doi.org/10.1007/s00213-022-06208-3
Parsons LH, Hurd YL (2015) Endocannabinoid signalling in reward and addiction. Nat Rev Neurosci 16(10):579–594. https://doi.org/10.1038/nrn4004
Rhodes JS, Best K, Belknap JK, Finn DA, Crabbe JC (2005) Evaluation of a simple model of ethanol drinking to intoxication in C57BL/6J mice. Physiol Behav 84(1):53–63. https://doi.org/10.1016/j.physbeh.2004.10.007
Salamone JD, McLaughlin PJ, Sink K, Makriyannis A, Parker LA (2007) Cannabinoid CB1 receptor inverse agonists and neutral antagonists: effects on food intake, food-reinforced behavior and food aversions. Physiol Behav 91(4):383–388. https://doi.org/10.1016/j.physbeh.2007.04.013
Serrano A, Natividad LA (2022) Alcohol-endocannabinoid interactions: implications for addiction-related behavioral processes. Alcohol Res 42(1):09. https://doi.org/10.35946/arcr.v42.1.09
Sink KS, McLaughlin PJ, Wood JA, Brown C, Fan P, Vemuri VK, Peng Y, Olszewska T, Thakur GA, Makriyannis A, Parker LA, Salamone JD (2008) The novel cannabinoid CB1 receptor neutral antagonist AM4113 suppresses food intake and food-reinforced behavior but does not induce signs of nausea in rats. Neuropsychopharmacology 33(4):946–955. https://doi.org/10.1038/sj.npp.1301476
Sink KS, Segovia KN, Sink J, Randall PA, Collins LE, Correa M, Markus EJ, Vemuri VK, Makriyannis A, Salamone JD (2010) Potential anxiogenic effects of cannabinoid CB1 receptor antagonists/inverse agonists in rats: comparisons between AM4113, AM251, and the benzodiazepine inverse agonist FG-7142. Eur Neuropsychopharmacol 20(2):112–122. https://doi.org/10.1016/j.euroneuro.2009.11.002
Sink KS, Vemuri VK, Wood J, Makriyannis A, Salamone JD (2009) Oral bioavailability of the novel cannabinoid CB1 antagonist AM6527: effects on food-reinforced behavior and comparisons with AM4113. Pharmacol Biochem Behav 91(3):303–306. https://doi.org/10.1016/j.pbb.2008.07.013
Sneddon EA, White RD, Radke AK (2019) Sex differences in binge-like and aversion-resistant alcohol drinking in C57BL/6J mice. Alcohol Clin Exp Res 43(2):243–249. https://doi.org/10.1111/acer.13923
Soyka M, Koller G, Schmidt P, Lesch OM, Leweke M, Fehr C, Gann H, Mann KF (2008) Cannabinoid receptor 1 blocker rimonabant (SR 141716) for treatment of alcohol dependence: results from a placebo-controlled, double-blind trial. J Clin Psychopharmacol 28(3):317–324. https://doi.org/10.1097/JCP.0b013e318172b8bc
Valenstein ES, Cox VC, Kakolewski JW (1967) Polydipsia elicited by the synergistic action of a saccharin and glucose solution. Science 157(3788):552–554. https://doi.org/10.1126/science.157.3788.552
Vemuri VK, Janero DR, Makriyannis A (2008) Pharmacotherapeutic targeting of the endocannabinoid signaling system: drugs for obesity and the metabolic syndrome. Physiol Behav 93(4-5):671–686. https://doi.org/10.1016/j.physbeh.2007.11.012
Wiley JL, Matthew Walentiny D, Vann RE, Baskfield CY (2011) Dissimilar cannabinoid substitution patterns in mice trained to discriminate Δ(9)-tetrahydrocannabinol or methanandamide from vehicle. Behav Pharmacol 22(5-6):480–488. https://doi.org/10.1097/FBP.0b013e328348eced
Witkiewitz K, Litten RZ, Leggio L (2019) Advances in the science and treatment of alcohol use disorder 5(9):eaax4043. https://doi.org/10.1126/sciadv.aax4043
Acknowledgements
The authors would like to thank J. Thomas Sopko for his excellent technical assistance.
Funding
This work was funded by NIH grants U01AA028963 (AM, PI), R01AA013983 (KAM) and R01DA031734 (KAM, PI), and R37DA023142 (AM).
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Raghav, J.G., Kumar, H., Ji, L. et al. The neutral CB1 antagonist AM6527 reduces ethanol seeking, binge-like consumption, reinforcing, and withdrawal effects in male and female mice. Psychopharmacology 241, 427–443 (2024). https://doi.org/10.1007/s00213-023-06500-w
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DOI: https://doi.org/10.1007/s00213-023-06500-w