Skip to main content
Log in

Inhibitory effect of intrathecally administered AM404, an endocannabinoid reuptake inhibitor, on neuropathic pain in a rat chronic constriction injury model

  • Article
  • Published:
Pharmacological Reports Aims and scope Submit manuscript

Abstract

Background

The endocannabinoid system modulates a wide variety of pain conditions. Systemically administered AM404, an endocannabinoid reuptake inhibitor, exerts antinociceptive effects via activation of the endocannabinoid system. However, the mechanism and site of AM404 action are not fully understood. Here, we explored the effect of AM404 on neuropathic pain at the site of the spinal cord.

Methods

Male Sprague–Dawley rats were subjected to chronic constriction injury (CCI) of the sciatic nerve. The effects of intrathecal administration of AM404 on mechanical and cold hyperalgesia were examined using the electronic von Frey test and cold plate test, respectively. Motor coordination was assessed using the rotarod test. To understand the mechanisms underlying the action of AM404, we tested the effects of pretreatment with the cannabinoid type 1 (CB1) receptor antagonist AM251, CB2 receptor antagonist AM630, and transient receptor potential vanilloid type 1 (TRPV1) antagonist capsazepine.

Results

AM404 attenuated mechanical and cold hyperalgesia with minimal effects on motor coordination. AM251 significantly inhibited the antihyperalgesic action of AM404, whereas capsazepine showed a potentiating effect.

Conclusions

These results indicate that AM404 exerts antihyperalgesic effects primarily via CB1, but not CB2, receptor activation at the site of the spinal cord. TRPV1 receptors appear to play a pronociceptive role in CCI rats. The endocannabinoid reuptake inhibitor may be a promising candidate treatment for neuropathic pain.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

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

Similar content being viewed by others

References

  1. Hill KP, Palastro MD, Johnson B, Ditre JW. Cannabis and pain: a clinical review. Cannabis Cannabinoid Res. 2017;2:96–104.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Guindon J, Hohmann AG. The endocannabinoid system and pain. CNS Neurol Disord Drug Targets. 2009;8:403–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Beltramo M, Stella N, Calignano A, Lin SY, Makriyannis A, Piomelli D. Functional role of high-affinity anandamide transport, as revealed by selective inhibition. Science. 1997;277:1094–7.

    Article  CAS  PubMed  Google Scholar 

  4. Calignano A, La Rana G, Beltramo M, Makriyannis A, Piomelli D. Potentiation of anandamide hypotension by the transport inhibitor, AM404. Eur J Pharmacol. 1997;337:R1-2.

    Article  CAS  PubMed  Google Scholar 

  5. Wiskerke J, Irimia C, Cravatt BF, De Vries TJ, Schoffelmeer ANM, Pattij T, et al. Characterization of the effects of reuptake and hydrolysis inhibition on interstitial endocannabinoid levels in the brain: an in vivo microdialysis study. ACS Chem Neurosci. 2012;3:407–17.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Giuffrida A, Rodríguez de Fonseca F, Nava F, Loubet-Lescoulié P, Piomelli D. Elevated circulating levels of anandamide after administration of the transport inhibitor, AM404. Eur J Pharmacol. 2000;408:161–8.

  7. Barrière DA, Boumezbeur F, Dalmann R, Cadeddu R, Richard D, Pinguet J, et al. Paracetamol is a centrally acting analgesic using mechanisms located in the periaqueductal grey. Br J Pharmacol. 2020;177:1773–92.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Klinger-Gratz PP, Ralvenius WT, Neumann E, Kato A, Nyilas R, Lele Z, et al. Acetaminophen relieves inflammatory pain through CB1 cannabinoid receptors in the rostral ventromedial medulla. J Neurosci. 2018;38:322–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Gühring H, Hamza M, Sergejeva M, Ates M, Kotalla CE, Ledent C, et al. A role for endocannabinoids in indomethacin-induced spinal antinociception. Eur J Pharmacol. 2002;454:153–63.

    Article  PubMed  Google Scholar 

  10. La Rana G, Russo R, Campolongo P, Bortolato M, Mangieri RA, Cuomo V, et al. Modulation of neuropathic and inflammatory pain by the endocannabinoid transport inhibitor AM404 [N-(4-hydroxyphenyl)-eicosa-5,8,11,14-tetraenamide]. J Pharmacol Exp Ther. 2006;317:1365–71.

    Article  PubMed  Google Scholar 

  11. Mitchell VA, Greenwood R, Jayamanne A, Vaughan CW. Actions of the endocannabinoid transport inhibitor AM404 in neuropathic and inflammatory pain models. Clin Exp Pharmacol Physiol. 2007;34:1186–90.

    CAS  PubMed  Google Scholar 

  12. Costa B, Siniscalco D, Trovato AE, Comelli F, Sotgiu ML, Colleoni M, et al. AM404, an inhibitor of anandamide uptake, prevents pain behavior and modulates cytokine and apoptotic pathways in a rat model of neuropathic pain. Br J Pharmacol. 2009;148:1022–32.

    Article  Google Scholar 

  13. Bennett GJ, Xie YK. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain. 1988;33:87–107.

    Article  PubMed  Google Scholar 

  14. Zajaczkowska R, Kwiatkowski K, Pawlik K, Piotrowska A, Rojewska E, Makuch W, et al. Metamizole relieves pain by influencing cytokine levels in dorsal root ganglia in a rat model of neuropathic pain. Pharmacol Rep. 2020;72:1310–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Zhong Y, Chen J, Chen J, Chen Y, Li L, Xie Y. Crosstalk between Cdk5/p35 and ERK1/2 signalling mediates spinal astrocyte activity via the PPARγ pathway in a rat model of chronic constriction injury. J Neurochem. 2019;151:166–84.

    Article  CAS  PubMed  Google Scholar 

  16. Hara K, Haranishi Y, Terada T. Intrathecally administered perampanel alleviates neuropathic and inflammatory pain in rats. Eur J Pharmacol. 2020;872:172949.

    Article  CAS  PubMed  Google Scholar 

  17. Haranishi Y, Hara K, Terada T. Antihyperalgesic effects of intrathecal perospirone in a rat model of neuropathic pain. Pharmacol Biochem Behav. 2020;195:172964.

    Article  CAS  PubMed  Google Scholar 

  18. Yang F, Xu Q, Shu B, Tiwari V, He SQ, Vera-Portocarrero LP, et al. Activation of cannabinoid CB1 receptor contributes to suppression of spinal nociceptive transmission and inhibition of mechanical hypersensitivity by Aβ-fiber stimulation. Pain. 2016;157:2582–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Landry RP, Martinez E, DeLeo JA, Romero-Sandoval EA. Spinal cannabinoid receptor type 2 agonist reduces mechanical allodynia and induces mitogen-activated protein kinase phosphatases in a rat model of neuropathic pain. J Pain. 2012;13:836–48.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Horvath G, Kekesi G, Nagy E, Benedek G. The role of TRPV1 receptors in the antinociceptive effect of anandamide at spinal level. Pain. 2008;134:277–84.

    Article  CAS  PubMed  Google Scholar 

  21. Woodhams SG, Chapman V, Finn DP, Hohmann AG, Neugebauer V. The cannabinoid system and pain. Neuropharmacology. 2017;124:105–20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Starowicz K, Przewlocka B. Modulation of neuropathic-pain-related behaviour by the spinal endocannabinoid/endovanilloid system. Philos Trans R Soc Lond B Biol Sci. 2012;367:3286–99.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Donvito G, Nass SR, Wilkerson JL, Curry ZA, Schurman LD, Kinsey SG, et al. The endogenous cannabinoid system: a budding source of targets for treating inflammatory and neuropathic pain. Neuropsychopharmacology. 2018;43:52–79.

    Article  CAS  PubMed  Google Scholar 

  24. Zhang J, Hoffert C, Vu HK, Groblewski T, Ahmad S, O’Donnell D. Induction of CB2 receptor expression in the rat spinal cord of neuropathic but not inflammatory chronic pain models. Eur J Neurosci. 2003;17:2750–4.

    Article  PubMed  Google Scholar 

  25. Wotherspoon G, Fox A, McIntyre P, Colley S, Bevan S, Winter J. Peripheral nerve injury induces cannabinoid receptor 2 protein expression in rat sensory neurons. Neuroscience. 2005;135:235–45.

    Article  CAS  PubMed  Google Scholar 

  26. Wilkerson JL, Gentry KR, Dengler EC, Wallace JA, Kerwin AA, Armijo LM, et al. Intrathecal cannabilactone CB(2)R agonist, AM1710, controls pathological pain and restores basal cytokine levels. Pain. 2012;153:1091–106.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Liu C, Walker JM. Effects of a cannabinoid agonist on spinal nociceptive neurons in a rodent model of neuropathic pain. J Neurophysiol. 2006;96:2984–94.

    Article  CAS  PubMed  Google Scholar 

  28. Brownjohn PW, Ashton JC. Spinal cannabinoid CB2 receptors as a target for neuropathic pain: an investigation using chronic constriction injury. Neuroscience. 2012;20:180–93.

    Article  Google Scholar 

  29. Rani Sagar D, Burston JJ, Woodhams SG, Chapman V. Dynamic changes to the endocannabinoid system in models of chronic pain. Philos Trans R Soc Lond B Biol Sci. 2012;367:3300–11.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Starowicz K, Finn DP. Cannabinoids and pain: sites and mechanisms of action. Adv Pharmacol. 2017;80:437–75.

    Article  CAS  PubMed  Google Scholar 

  31. Chen Y, Willcockson HH, Valtschanoff JG. Influence of the vanilloid receptor TRPV1 on the activation of spinal cord glia in mouse models of pain. Exp Neurol. 2009;220:383–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Talbot S, Dias JP, Lahjouji K, Bogo MR, Campos MM, Gaudreau P, et al. Activation of TRPV1 by capsaicin induces functional kinin B(1) receptor in rat spinal cord microglia. J Neuroinflammation. 2012;9:16.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Gunthorpe MJ, Chizh BA. Clinical development of TRPV1 antagonists: targeting a pivotal point in the pain pathway. Drug Discov Today. 2009;14:56–67.

    Article  CAS  PubMed  Google Scholar 

  34. Wong GY, Gavva NR. Therapeutic potential of vanilloid receptor TRPV1 agonists and antagonists as analgesics: recent advances and setbacks. Brain Res Rev. 2009;60:267–77.

    Article  CAS  PubMed  Google Scholar 

  35. Spicarova D, Palecek J. The role of the TRPV1 endogenous agonist N-Oleoyldopamine in modulation of nociceptive signaling at the spinal cord level. J Neurophysiol. 2009;102:234–43.

    Article  CAS  PubMed  Google Scholar 

  36. Kim YH, Back SK, Davies AJ, Jeong H, Jo HJ, Chung G, et al. TRPV1 in GABAergic interneurons mediates neuropathic mechanical allodynia and disinhibition of the nociceptive circuitry in the spinal cord. Neuron. 2012;74:640–7.

    Article  CAS  PubMed  Google Scholar 

  37. Watabiki T, Kiso T, Tsukamoto M, Aoki T, Matsuoka N. Intrathecal administration of AS1928370, a transient receptor potential vanilloid 1 antagonist, attenuates mechanical allodynia in a mouse model of neuropathic pain. Biol Pharm Bull. 2011;34:1105–8.

    Article  CAS  PubMed  Google Scholar 

  38. Maione S, De Petrocellis L, de Novellis V, Moriello AS, Petrosino S, Palazzo E, et al. Analgesic actions of N-arachidonoyl-serotonin, a fatty acid amide hydrolase inhibitor with antagonistic activity at vanilloid TRPV1 receptors. Br J Pharmacol. 2007;150:766–81.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Malek N, Starowicz K. Dual-acting compounds targeting endocannabinoid and endovanilloid systems-a novel treatment option for chronic pain management. Front Pharmacol. 2016;7:257.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Högestätt ED, Jönsson BA, Ermund A, Andersson DA, Björk H, Alexander JP, et al. Conversion of acetaminophen to the bioactive N-acylphenolamine AM404 via fatty acid amide hydrolase-dependent arachidonic acid conjugation in the nervous system. J Biol Chem. 2005;280:31405–12.

    Article  PubMed  Google Scholar 

  41. Saliba SW, Marcotegui AR, Fortwängler E, Ditrich J, Perazzo JC, Muñoz E, et al. AM404, paracetamol metabolite, prevents prostaglandin synthesis in activated microglia by inhibiting COX activity. J Neuroinflammation. 2017;14:246.

    Article  PubMed  PubMed Central  Google Scholar 

  42. Alptekin A, Galadari S, Shuba Y, Petroianu G, Oz M. The effects of anandamide transport inhibitor AM404 on voltage-dependent calcium channels. Eur J Pharmacol. 2010;634:10–5.

    Article  CAS  PubMed  Google Scholar 

  43. Nicholson RA, Liao C, Zheng J, David LS, Coyne L, Errington AC, et al. Sodium channel inhibition by anandamide and synthetic cannabimimetics in brain. Brain Res. 2008;978:194–204.

    Article  Google Scholar 

  44. Barann M, Molderings G, Brüss M, Bönisch H, Urban BW, Göthert M. Direct inhibition by cannabinoids of human 5-HT3A receptors: probable involvement of an allosteric modulatory site. Br J Pharmacol. 2002;137:589–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Hejazi N, Zhou C, Oz M, Sun H, Ye JH, Zhang L. Delta9-tetrahydrocannabinol and endogenous cannabinoid anandamide directly potentiate the function of glycine receptors. Mol Pharmacol. 2006;69:991–7.

    Article  CAS  PubMed  Google Scholar 

  46. Chen SR, Eisenach JC, McCaslin PP, Pan HL. Synergistic effect between intrathecal non-NMDA antagonist and gabapentin on allodynia induced by spinal nerve ligation in rats. Anesthesiology. 2000;92:500–6.

    Article  CAS  PubMed  Google Scholar 

  47. Yamama Y, Nishikawa K, Funao T, Mori T, Asada A. Intrathecal gabapentin and clonidine synergistically inhibit allodynia in spinal nerve-ligated rats. Life Sci. 2010;87:565–71.

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This work was supported by JSPS KAKENHI Grant Number JP18K08838 (KH).

Author information

Authors and Affiliations

Authors

Contributions

YH contributed to the conception of the study, the acquisition, analysis and interpretation of data for the study, and drafting the manuscript. KH contributed to the design of the study, the acquisition and interpretation of data for the study, and revising the manuscript critically for important intellectual content. TT contributed to the acquisition and interpretation of data for the study and approved final manuscript.

Corresponding author

Correspondence to Koji Hara.

Ethics declarations

Conflict of interest

Authors have nothing to disclose.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Haranishi, Y., Hara, K. & Terada, T. Inhibitory effect of intrathecally administered AM404, an endocannabinoid reuptake inhibitor, on neuropathic pain in a rat chronic constriction injury model. Pharmacol. Rep 73, 820–827 (2021). https://doi.org/10.1007/s43440-021-00250-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s43440-021-00250-2

Keywords

Navigation