Inhibitors of Endocannabinoid-Metabolizing Enzymes Reduce Precipitated Withdrawal Responses in THC-Dependent Mice

Abstract

Abstinence symptoms in cannabis-dependent individuals are believed to contribute to the maintenance of regular marijuana use. However, there are currently no medications approved by the FDA to treat cannabis-related disorders. The only treatment currently shown consistently to alleviate cannabinoid withdrawal in both animals and humans is substitution therapy using the psychoactive constituent of marijuana, Δ9-tetrahydrocannabinol (THC). However, new genetic and pharmacological tools are available to increase endocannabinoid levels by targeting fatty acid amide hydrolase (FAAH) or monoacylglycerol lipase (MAGL), the enzymes responsible for the degradation of the endogenous cannabinoid ligands anandamide and 2-arachidonoylglycerol, respectively. In the present study, we investigated whether increasing endogenous cannabinoids levels, through the use of FAAH (−/−) mice as well as the FAAH inhibitor URB597 or the MAGL inhibitor JZL184, would reduce the intensity of withdrawal signs precipitated by the CB1 receptor antagonist rimonabant in THC-dependent mice. Strikingly, acute administration of either URB597 or JZL184 significantly attenuated rimonabant-precipitated withdrawal signs in THC-dependent mice. In contrast, FAAH (−/−) mice showed identical withdrawal responses as wild-type mice under a variety of conditions, suggesting that the absence of this enzyme across the development of dependence and during rimonabant challenge does not affect withdrawal responses. Of importance, subchronic administration of URB597 did not lead to cannabinoid dependence and neither URB597 nor JZL184 impaired rotarod motor coordination. These results support the concept of targeting endocannabinoid metabolizing enzymes as a promising treatment for cannabis withdrawal.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Abbreviations

FAAH:

Fatty acid amide hydrolase

Rimonabant:

N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide HCl

THC:

{Delta}9-tetrahydrocannabinol

URB597:

Cyclohexylcarbamic acid 3′-carbamoylbiphenyl-3-yl ester

C57:

C57BL/6J mouse strain

JZL184:

4-Nitrophenyl 4-(dibenzo[d][1,3]dioxol-5-yl(hydroxy)methyl)piperidine-1-carboxylate

MAGL:

Monoacylglycerol lipase

AEA:

Anandamide

2-AG:

2-Arachindonoylglycerol

References

  1. 1.

    Substance Abuse and Mental Health Services Administration: Office of Applied Studies. Results from the 2007 National Survey on Drug Use and Health: national findings. Rockville, MD: Dept. of Health and Human Services, Substance Abuse and Mental Health Services Administration, Office of Applied Studies; 2008.

  2. 2.

    Budney AJ, Moore BA, Vandrey RG, Hughes JR. The time course and significance of cannabis withdrawal. J Abnorm Psychol 2003;112(3):393–402.

    PubMed  Article  Google Scholar 

  3. 3.

    Haney M, Ward AS, Comer SD, Foltin RW, Fischman MW. Abstinence symptoms following smoked marijuana in humans. Psychopharmacology (Berl) 1999;141(4):395–404.

    Article  CAS  Google Scholar 

  4. 4.

    Budney AJ, Vandrey RG, Hughes JR, Thostenson JD, Bursac Z. Comparison of cannabis and tobacco withdrawal: severity and contribution to relapse. J Subst Abuse Treat 2008;35(4):362–8.

    PubMed  Article  Google Scholar 

  5. 5.

    Vandrey RG, Budney AJ, Hughes JR, Liguori A. A within-subject comparison of withdrawal symptoms during abstinence from cannabis, tobacco, and both substances. Drug Alcohol Depend 2008;92(1–3):48–54.

    PubMed  Article  CAS  Google Scholar 

  6. 6.

    Haney M, Ward AS, Comer SD, Foltin RW, Fischman MW. Abstinence symptoms following oral THC administration to humans. Psychopharmacology (Berl) 1999;141(4):385–94.

    Article  CAS  Google Scholar 

  7. 7.

    Lichtman AH, Fisher J, Martin BR. Precipitated cannabinoid withdrawal is reversed by Delta(9)-tetrahydrocannabinol or clonidine. Pharmacol Biochem Behav 2001;69(1–2):181–8.

    PubMed  Article  CAS  Google Scholar 

  8. 8.

    Wilson DM, Varvel SA, Harloe JP, Martin BR, Lichtman AH. SR 141716 (Rimonabant) precipitates withdrawal in marijuana-dependent mice. Pharmacol Biochem Behav 2006;85(1):105–13.

    PubMed  Article  CAS  Google Scholar 

  9. 9.

    Beardsley PM, Balster RL, Harris LS. Dependence on tetrahydrocannabinol in rhesus monkeys. J Pharmacol Exp Ther 1986;239(2):311–9.

    PubMed  CAS  Google Scholar 

  10. 10.

    Haney M, Hart CL, Vosburg SK, Nasser J, Bennett A, Zubaran C, et al. Marijuana withdrawal in humans: effects of oral THC or divalproex. Neuropsychopharmacology 2004;29(1):158–70.

    PubMed  Article  CAS  Google Scholar 

  11. 11.

    Budney AJ, Vandrey RG, Hughes JR, Moore BA, Bahrenburg B. Oral delta-9-tetrahydrocannabinol suppresses cannabis withdrawal symptoms. Drug Alcohol Depend 2007;86(1):22–9.

    PubMed  Article  CAS  Google Scholar 

  12. 12.

    Haney M, Ward AS, Comer SD, Hart CL, Foltin RW, Fischman MW. Bupropion SR worsens mood during marijuana withdrawal in humans. Psychopharmacology (Berl) 2001;155(2):171–9.

    Article  CAS  Google Scholar 

  13. 13.

    Aceto MD, Scates SM, Lowe JA, Martin BR. Cannabinoid precipitated withdrawal by the selective cannabinoid receptor antagonist, SR 141716A. Eur J Pharmacol 1995;282(1–3):R1–2.

    PubMed  Article  CAS  Google Scholar 

  14. 14.

    Tsou K, Patrick SL, Walker JM. Physical withdrawal in rats tolerant to delta 9-tetrahydrocannabinol precipitated by a cannabinoid receptor antagonist. Eur J Pharmacol 1995;280(3):R13–5.

    PubMed  Article  CAS  Google Scholar 

  15. 15.

    Cook SA, Lowe JA, Martin BR. CB1 receptor antagonist precipitates withdrawal in mice exposed to Delta9-tetrahydrocannabinol. J Pharmacol Exp Ther 1998;285(3):1150–6.

    PubMed  CAS  Google Scholar 

  16. 16.

    Hutcheson DM, Tzavara ET, Smadja C, Valjent E, Roques BP, Hanoune J, et al. Behavioural and biochemical evidence for signs of abstinence in mice chronically treated with delta-9-tetrahydrocannabinol. Br J Pharmacol 1998;125(7):1567–77.

    PubMed  Article  CAS  Google Scholar 

  17. 17.

    Tzavara ET, Valjent E, Firmo C, Mas M, Beslot F, Defer N, et al. Cannabinoid withdrawal is dependent upon PKA activation in the cerebellum. Eur J Neurosci 2000;12(3):1038–46.

    PubMed  Article  CAS  Google Scholar 

  18. 18.

    Howlett AC, Qualy JM, Khachatrian LL. Involvement of Gi in the inhibition of adenylate cyclase by cannabimimetic drugs. Mol Pharmacol 1986;29(3):307–13.

    PubMed  CAS  Google Scholar 

  19. 19.

    Sim-Selley LJ, Martin BR. Effect of chronic administration of R-(+)-[2,3-Dihydro-5-methyl-3-[(morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-b enzoxazinyl]-(1-naphthalenyl)methanone mesylate (WIN55,212–2) or delta(9)-tetrahydrocannabinol on cannabinoid receptor adaptation in mice. J Pharmacol Exp Ther 2002;303(1):36–44.

    PubMed  Article  CAS  Google Scholar 

  20. 20.

    Justinova Z, Tanda G, Redhi GH, Goldberg SR. Self-administration of delta9-tetrahydrocannabinol (THC) by drug naive squirrel monkeys. Psychopharmacology (Berl) 2003;169(2):135–40.

    Article  CAS  Google Scholar 

  21. 21.

    Jarbe TU, Henriksson BG. Discriminative response control produced with hashish, tetrahydrocannabinols (delta 8-THC and delta 9-THC), and other drugs. Psychopharmacologia 1974;40(1):1–16.

    PubMed  Article  CAS  Google Scholar 

  22. 22.

    Clapper JR, Mangieri R, Piomelli D. The endocannabinoid system as a target for the treatment of cannabis dependence. Neuropharmacology 2009;56(S1):235–43.

    PubMed  Article  CAS  Google Scholar 

  23. 23.

    Cravatt BF, Demarest K, Patricelli MP, Bracey MH, Giang DK, Martin BR, et al. Supersensitivity to anandamide and enhanced endogenous cannabinoid signaling in mice lacking fatty acid amide hydrolase. Proc Natl Acad Sci U S A 2001;98(16):9371–6.

    PubMed  Article  CAS  Google Scholar 

  24. 24.

    Fegley D, Gaetani S, Duranti A, Tontini A, Mor M, Tarzia G, et al. Characterization of the fatty acid amide hydrolase inhibitor cyclohexyl carbamic acid 3′-carbamoyl-biphenyl-3-yl ester (URB597): effects on anandamide and oleoylethanolamide deactivation. J Pharmacol Exp Ther 2005;313(1):352–8.

    PubMed  Article  CAS  Google Scholar 

  25. 25.

    Lichtman AH, Hawkins EG, Griffin G, Cravatt BF. Pharmacological activity of fatty acid amides is regulated, but not mediated, by fatty acid amide hydrolase in vivo. J Pharmacol Exp Ther 2002;302(1):73–9.

    PubMed  Article  CAS  Google Scholar 

  26. 26.

    Lichtman AH, Shelton CC, Advani T, Cravatt BF. Mice lacking fatty acid amide hydrolase exhibit a cannabinoid receptor-mediated phenotypic hypoalgesia. Pain 2004;109(3):319–27.

    PubMed  Article  CAS  Google Scholar 

  27. 27.

    Boger DL, Miyauchi H, Du W, Hardouin C, Fecik RA, Cheng H, et al. Discovery of a potent, selective, and efficacious class of reversible alpha-ketoheterocycle inhibitors of fatty acid amide hydrolase effective as analgesics. J Med Chem 2005;48(6):1849–56.

    PubMed  Article  CAS  Google Scholar 

  28. 28.

    Piomelli D, Tarzia G, Duranti A, Tontini A, Mor M, Compton TR, et al. Pharmacological profile of the selective FAAH inhibitor KDS-4103 (URB597). CNS Drug Rev 2006;12(1):21–38.

    PubMed  Article  CAS  Google Scholar 

  29. 29.

    Justinova Z, Mangieri RA, Bortolato M, Chefer SI, Mukhin AG, Clapper JR, et al. Fatty acid amide hydrolase inhibition heightens anandamide signaling without producing reinforcing effects in primates. Biol Psychiatry 2008;64(11):930–7.

    PubMed  Article  CAS  Google Scholar 

  30. 30.

    Solinas M, Justinova Z, Goldberg SR, Tanda G. Anandamide administration alone and after inhibition of fatty acid amide hydrolase (FAAH) increases dopamine levels in the nucleus accumbens shell in rats. J Neurochem 2006;98(2):408–19.

    PubMed  Article  CAS  Google Scholar 

  31. 31.

    Solinas M, Tanda G, Justinova Z, Wertheim CE, Yasar S, Piomelli D, et al. The endogenous cannabinoid anandamide produces delta-9-tetrahydrocannabinol-like discriminative and neurochemical effects that are enhanced by inhibition of fatty acid amide hydrolase but not by inhibition of anandamide transport. J Pharmacol Exp Ther 2007;321(1):370–80.

    PubMed  Article  CAS  Google Scholar 

  32. 32.

    Blankman JL, Simon GM, Cravatt BF. A comprehensive profile of brain enzymes that hydrolyze the endocannabinoid 2-arachidonoylglycerol. Chem Biol 2007;14(12):1347–56.

    PubMed  Article  CAS  Google Scholar 

  33. 33.

    Long JZ, Li W, Booker L, Burston JJ, Kinsey SG, Schlosburg JE, et al. Selective blockade of 2-arachidonoylglycerol hydrolysis produces cannabinoid behavioral effects. Nat Chem Biol 2009;5(1):37–44.

    PubMed  Article  CAS  Google Scholar 

  34. 34.

    Kathuria S, Gaetani S, Fegley D, Valino F, Duranti A, Tontini A, et al. Modulation of anxiety through blockade of anandamide hydrolysis. Nat Med 2003;9(1):76–81.

    PubMed  Article  CAS  Google Scholar 

  35. 35.

    Gobbi G, Bambico FR, Mangieri R, Bortolato M, Campolongo P, Solinas M, et al. Antidepressant-like activity and modulation of brain monoaminergic transmission by blockade of anandamide hydrolysis. Proc Natl Acad Sci U S A 2005;102(51):18620–5.

    PubMed  Article  CAS  Google Scholar 

  36. 36.

    Gonzalez S, Fernandez-Ruiz J, Di Marzo V, Hernandez M, Arevalo C, Nicanor C, et al. Behavioral and molecular changes elicited by acute administration of SR141716 to Delta9-tetrahydrocannabinol-tolerant rats: an experimental model of cannabinoid abstinence. Drug Alcohol Depend 2004;74(2):159–70.

    PubMed  Article  CAS  Google Scholar 

  37. 37.

    Darmani NA, Pandya DK. Involvement of other neurotransmitters in behaviors induced by the cannabinoid CB1 receptor antagonist SR 141716A in naive mice. J Neural Transm 2000;107(8–9):931–45.

    PubMed  Article  CAS  Google Scholar 

  38. 38.

    Janoyan JJ, Crim JL, Darmani NA. Reversal of SR 141716A-induced head-twitch and ear-scratch responses in mice by delta 9-THC and other cannabinoids. Pharmacol Biochem Behav 2002;71(1–2):155–62.

    PubMed  Article  CAS  Google Scholar 

  39. 39.

    Schlosburg JE, Boger DL, Lichtman AH. Endocannabinoid modulation of scratching response in an acute allergenic model: a new prospective neural therapeutic target for pruritus. J Pharmacol Exp Ther 2009;329(1):314–23.

    PubMed  Article  CAS  Google Scholar 

  40. 40.

    Hohmann AG, Suplita RL, Bolton NM, Neely MH, Fegley D, Mangieri R, et al. An endocannabinoid mechanism for stress-induced analgesia. Nature 2005;435(7045):1108–12.

    PubMed  Article  CAS  Google Scholar 

  41. 41.

    Saghatelian A, McKinney MK, Bandell M, Patapoutian A, Cravatt BF. A FAAH-regulated class of N-acyl taurines that activates TRP ion channels. Biochemistry 2006;45(30):9007–15.

    PubMed  Article  CAS  Google Scholar 

  42. 42.

    Bisogno T, Berrendero F, Ambrosino G, Cebeira M, Ramos JA, Fernandez-Ruiz JJ, et al. Brain regional distribution of endocannabinoids: implications for their biosynthesis and biological function. Biochem Biophys Res Commun 1999;256(2):377–80.

    PubMed  Article  CAS  Google Scholar 

  43. 43.

    Crowley TJ. Adolescents and substance-related disorders: research agenda to guide decisions on Diagnostic and Statistical Manual of Mental Disorders, fifth edition (DSM-V). Addiction 2006;101(Suppl 1):115–24.

    PubMed  Article  Google Scholar 

Download references

Acknowledgements

The authors would like to acknowledge the technical assistance of Noor S. Shubar Ali, Deborah Karp, and Megan O’Brien with rotarod testing. The work was supported by National Institute on Drug Abuse grants P01DA017259, R01DA15197, R01DA03672, R01DA02396, R01DA015683, P50DA005274, P01DA009789 and T32DA007027. Additional support was provided by Scholar Rescue Funds of The Institute of International Education, New York.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Aron H. Lichtman.

Additional information

Guest Editor: Rao Rapaka

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Schlosburg, J.E., Carlson, B.L.A., Ramesh, D. et al. Inhibitors of Endocannabinoid-Metabolizing Enzymes Reduce Precipitated Withdrawal Responses in THC-Dependent Mice. AAPS J 11, 342–352 (2009). https://doi.org/10.1208/s12248-009-9110-7

Download citation

Key words

  • anandamide
  • cannabis dependence
  • fatty acid amide hydrolase (FAAH)
  • monoacylglycerol lipase (MAGL)
  • 2-arachindonoylglycerol (2-AG)