Metabolism of synthetic cannabinoids PB-22 and its 5-fluoro analog, 5F-PB-22, by human hepatocyte incubation and high-resolution mass spectrometry
- 1.3k Downloads
PB-22 (1-pentyl-8-quinolinyl ester-1H-indole-3-carboxylic acid) and 5F-PB-22 (1-(5-fluoropentyl)-8-quinolinyl ester-1H-indole-3-carboxylic acid) are new synthetic cannabinoids with a quinoline substructure and the first marketed substances with an ester bond linkage. No human metabolism data are currently available, making it difficult to document PB-22 and 5F-PB-22 intake from urine analysis, and complicating assessment of the drugs’ pharmacodynamic and toxicological properties.
We incubated 10 μmol/l PB-22 and 5F-PB-22 with pooled cryopreserved human hepatocytes up to 3 h and analyzed samples on a TripleTOF 5600+ high-resolution mass spectrometer. Data were acquired via TOF scan, followed by information-dependent acquisition triggered product ion scans with mass defect filtering (MDF). The accurate mass full scan MS and MS/MS metabolite datasets were analyzed with multiple data processing techniques, including MDF, neutral loss and product ion filtering.
The predominant metabolic pathway for PB-22 and 5F-PB-22 was ester hydrolysis yielding a wide variety of (5-fluoro)pentylindole-3-carboxylic acid metabolites. Twenty metabolites for PB-22 and 22 metabolites for 5F-PB-22 were identified, with the majority generated by oxidation with or without glucuronidation. For 5F-PB-22, oxidative defluorination occurred forming PB-22 metabolites. Both compounds underwent epoxide formation followed by internal hydrolysis and also produced a cysteine conjugate.
Human hepatic metabolic profiles were generated for PB-22 and 5F-PB-22. Pentylindole-3-carboxylic acid, hydroxypentyl-PB-22 and PB-22 pentanoic acid for PB-22, and 5′-fluoropentylindole-3-carboxylic acid, PB-22 pentanoic acid and the hydroxy-5F-PB-22 metabolite with oxidation at the quinoline system for 5F-PB-22 are likely the best targets to incorporate into analytical methods for urine to document PB-22 and 5F-PB-22 intake.
KeywordsSynthetic cannabinoids PB-22 5F-PB-22 High resolution mass spectrometry Metabolism
This research was funded by the Intramural Research Program of the National Institute on Drug Abuse, National Institutes of Health and AB SCIEX.
- 1.Sedefov R, Gallegos A, Kind L, Lopez D, Auwarter V, Hughes B. EMCCDA 2009 Thematic paper - Understanding the ‘Spice’ phenomenon. Office for Official Publications of the European Communities, 2009Google Scholar
- 5.Perrone D, Helgesen RD, Fischer RG (2013) United States drug prohibition and legal highs: How drug testing may lead cannabis users to spice. Drugs: Education Prevention Policy 20:216–24Google Scholar
- 6.European Monitoring Centre for Drugs and Drug Abuse. European Database on New Drugs. https://ednd-cma.emcdda.europa.eu/ (Accessed Sept. 2013).
- 7.NMS Labs. Designer drug trends report. NMS Labs, 2013Google Scholar
- 8.US Government. Synthetic Drug Abuse Prevention Act of 2012 (S.3187), 2012Google Scholar
- 9.Uchiyama N, Kawamura M, Kikura-Hanajiri R, Goda Y (2012) Identification of two new-type synthetic cannabinoids, N-(1-adamantyl)-1-pentyl-1H-indole-3-carboxamide (APICA) and N-(1-adamantyl)-1-pentyl-1H-indazole-3-carboxamide (APINACA), and detection of five synthetic cannabinoids, AM-1220, AM-2233, AM-1241, CB-13 (CRA-13), and AM-1248, as designer drugs in illegal products. Forensic Toxicol 30:114–25CrossRefGoogle Scholar
- 11.Uchiyama N, Matsuda S, Kawamura M, Kikura-Hanajiri R, Goda Y (2013) Two new-type cannabimimetic quinolinyl carboxylates, QUPIC and QUCHIC, two new cannabimimetic carboxamide derivatives, ADB-FUBINACA and ADBICA, and five synthetic cannabinoids detected with a thiophene derivative α-PVT and an opioid receptor agonist AH-7921 identified in illegal products. Forensic Toxicol 31:223–40CrossRefGoogle Scholar
- 12.Department of Justice, Drug Enforcement Administration. Schedules of controlled substances: Temporary placement of three synthetic cannabinoids into schedule I. Federal Register, Vol. 78, 2013.Google Scholar
- 13.Uchiyama N, Matsuda S, Kawamura M, Kikura-Hanajiri R, Goda Y Identification of two new-type designer drugs, piperazine derivative MT-45 (I-C6) and synthetic peptide noopept (GVS-111), with synthetic cannabinoid A-834735, cathinone derivative 4-methoxy-α-PVP, and phenethylamine derivative 4-methylbuphedrine from illegal products. Forensic Toxicol. doi: 10.1007/s11419-013-0194-5
- 14.Martin T. Report about the number of exhibits containing synthetic cannabinoids from October 2011 till May 2013 analyzed by the Drug Chemistry Branch, USACIL. Personal communication, Castaneto M, 2013.Google Scholar
- 15.Berrier A (2013) Classes and structures of emerging cannabimimetics and cathinones. Emerging Trends in Synthetic Drugs, Workshop, Gaithersburg, MD, USAGoogle Scholar
- 17.Reddit Drug User Forum. My PB-22 experience. http://www.reddit.com/r/Drugs/comments/1b4mb8/my_pb22_experience/ (Accessed August 27, 2013).
- 18.DrugsForum. 5 F-PB-22 drug info. http://www.drugs-forum.com/forum/showthread.php?t=202998 (Accessed August 27, 2013).
- 20.Wohlfarth A, Pang S, Zhu M, Gandhi AS, Scheidweiler KB (2013) Liu H.-F., Huestis MA. First metabolic profile of XLR-11, a novel synthetic cannabinoid, obtained by using human hepatocytes and high-resolution mass spectrometry. Clin Chem 59:1638–48Google Scholar
- 21.Grigoryev A, Savchuk S, Melnik A, Moskaleva N, Dzhurko J, Ershov M et al (2011) Chromatography-mass spectrometry studies on the metabolism of synthetic cannabinoids JWH-018 and JWH-073, psychoactive components of smoking mixtures. J Chromatogr B Analyt Technol Biomed Life Sci 879:1126–36CrossRefGoogle Scholar
- 24.Gandhi A, Zhu M, Pang S, Wohlfarth A, Scheidweiler K, Liu H-f, Huestis M. First characterization of AKB-48 metabolism, a novel synthetic cannabinoid, using human hepatocytes and high-resolution mass spectrometry. AAPS J 2013:1-8.Google Scholar
- 25.Horng H, Benet LZ. The non-enzymatic reactivity of the acyl-linked metabolites of mefenamic acid towards amino and thiol functional group biomolecules. Drug Metabolism and Disposition 2013Google Scholar
- 26.Andersen. Cosmetic ingredient review: Final amended report on the safety assessment of oxyquinoline and oxyquinoline sulfate as used in cosmetics. Int J Toxicol 2006;25:1-9.Google Scholar
- 27.Gershon H, Parmegiani R (1963) Antimicrobial activity of 8-quinolinol, its salts with salicylic acid and 3-hydroxy-2-naphthoic acid, and the respective copper (II) chelates in liquid culture. Appl Microbiol 11:62–5Google Scholar
- 28.Youatt J (1982) Oxine, ferric oxine and copper oxine as inhibitors of growth and differentiation of allomyces macrogynus. Aust J Biol Sci 35:565–72Google Scholar
- 29.National Toxicology Program. NTP technical report on the toxicology and carcinogenesis studies of 8-hydroxyquinoline in F344/N rats and B6C3F1 mice (feed studies). NTP TR 276, NIH Publication No 85-2532, 1974Google Scholar
- 30.National Library of Medicine Database. Indium-11-oxyquinoline solution. National Library of Medicine, 2013Google Scholar
- 34.Grillo MP, Knutson CG, Sanders PE, Waldon DJ, Hua F, Ware JA (2003) Studies on the chemical reactivity of diclofenac acyl glucuronide with glutathione: Identification of diclofena-s-acyl-glutathione in rat bile. Drug Metab Dispos 31:1327–36Google Scholar
- 35.Shore LJ, Fenselau C, King AR, Dickinson RG (1995) Characterization and formation of the glutathione conjugate of clofibric acid. Drug Metab Dispos 23:119–23Google Scholar
- 36.Cayman Chemical. www.caymanchem.com (Accessed November 2013).