Skip to main content

Advertisement

SpringerLink
Log in

Diagnosing intake and rationalizing toxicities associated with 5F-MDMB-PINACA and 4F-MDMB-BINACA abuse

  • Toxicokinetics and Metabolism
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

5F-MDMB-PINACA and 4F-MDMB-BINACA are synthetic cannabinoids (SCs) that elicit cannabinoid psychoactive effects. Defining pharmacokinetic–pharmacodynamic (PK–PD) relationships governing SCs and their metabolites are paramount to investigating their in vivo toxicological outcomes. However, the disposition kinetics and cannabinoid receptor (CB) activities of the primary metabolites of SCs are largely unknown. Additionally, reasons underlying the selection of ester hydrolysis metabolites (EHMs) as urinary biomarkers are often unclear. Here, metabolic reaction phenotyping was performed to identify key metabolizing enzymes of the parent SCs. Hepatic clearances of parent SCs and their EHMs were estimated from microsomal metabolic stability studies. Renal clearances were simulated using a mechanistic kidney model incorporating in vitro permeability and organic anionic transporter 3 (OAT3)-mediated uptake data. Overall clearances were considered in tandem with estimated volumes of distribution for in vivo biological half-lives (t1/2) predictions. Interactions of the compounds with CB1 and CB2 were investigated using a G-protein coupled receptor activation assay. We demonstrated that similar enzymatic isoforms were implicated in the metabolism of 5F-MDMB-PINACA and 4F-MDMB-BINACA. Our in vivo t1/2 determinations verified the rapid elimination of parent SCs and suggest prolonged circulation of their EHMs. The pronounced attenuation of the potencies and efficacies of the metabolites against CB1 and CB2 further suggests how toxic manifestations of SC abuse are likely precipitated by augmented exposure to parent SCs. Notably, basolateral OAT3-mediated uptake of the EHMs substantiates their higher urinary abundance. These novel insights underscore the importance of mechanistic, quantitative and systematic characterization of PK–PD relationships in rationalizing the toxicities of SCs.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

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

Similar content being viewed by others

Abbreviations

AUCm/AUCp :

Relative area under the curve of metabolite to parent

BNPP:

Bis(4-nitrophenyl) phosphate

CB:

Cannabinoid receptor

C b/C :

Ratio of drug concentration in blood to plasma

CLbsl,scr :

In vivo basolateral secretion clearance

CLformation :

In vivo formation clearance of the ester hydrolysis metabolite

CLH :

In vivo hepatic clearance

CLint,active :

In vitro intrinsic transporter-mediated active uptake

CLint,passive :

In vitro intrinsic passive uptake

CLint,formation :

In vitro intrinsic formation clearance of the ester hydrolysis metabolite

CLint,metabolic :

In vitro intrinsic metabolic clearance of the ester hydrolysis metabolite

CLint,total :

In vitro total intrinsic clearance of the parent synthetic cannabinoid

CLR :

In vivo renal clearance

CYP450:

Cytochrome P450 enzyme

CES:

Carboxylesterase

DDI:

Drug–drug interaction

EC50 :

Concentration of agonist that provokes a response halfway between the baseline and maximum response

EHM:

Ester hydrolysis metabolite

E max :

The maximal limit of response to an agonist

f u :

Fraction unbound of drug in plasma

f ub :

Fraction unbound of drug in blood

4F-MDMB-BINACA:

Methyl 2‐[1-(4‐fluorobutyl)‐1H‐indazole‐3‐carboxamido]‐3,3-dimethylbutanoate

5F-MDMB-PINACA:

Methyl 2‐[1-(5‐fluoropentyl)‐1H‐indazole‐3‐carboxamido]‐3,3‐dimethylbutanoate

HEK:

Human embryonic kidney 293

HLM:

Human liver microsomes

ISEF:

Intersystem extrapolation factor

IVIVE:

In vitro in vivo extrapolation

k :

Elimination rate constant

MDCK II:

Madin–Darby Canine Kidney II

nsSNP:

Non-synonymous small nucleotide polymorphism

OAT:

Organic anion transporter

P app :

Apparent permeability

pK a :

Acid dissociation constant

rhCYP450:

Recombinant human cytochrome P450 enzyme

rhCES:

Recombinant human carboxylesterase

SC:

Synthetic cannabinoid

t 1/2 :

Half-life

UGT:

Uridine 5′-diphospho-glucuronosyltransferase

USFDA:

United States Food and Drug Administration

V ss :

Volume of distribution at steady state

References

  • Antonides LH, Cannaert A, Norman C et al (2019) Enantiospecific synthesis, chiral separation, and biological activity of four indazole-3-carboxamide-type synthetic cannabinoid receptor agonists and their detection in seized drug samples. Front Chem 7:1–20

    Article  Google Scholar 

  • Cannaert A, Storme J, Franz F et al (2016) Detection and activity profiling of synthetic cannabinoids and metabolites with a newly developed bio-assay. Anal Chem 88:11476–11485

    Article  CAS  Google Scholar 

  • Cannaert A, Franz F, Auwärter V, Stove CP (2017) Activity-based detection of consumption of synthetic cannabinoids in authentic urine samples using a stable cannabinoid reporter system. Anal Chem 89:9527–9536

    Article  CAS  Google Scholar 

  • Castaneto MS, Wohlfarth A, Desrosiers NA et al (2015) Synthetic cannabinoids pharmacokinetics and detection methods in biological matrices. Drug Metab Rev 47:124–174

    Article  CAS  Google Scholar 

  • Chen Y, Liu L, Nguyen K, Fretland AJ (2011) Utility of intersystem extrapolation factors in early reaction phenotyping and the quantitative extrapolation of human liver microsomal intrinsic clearance using recombinant cytochromes P450. Drug Metab Dispos 39:373–382

    Article  CAS  Google Scholar 

  • Diao X, Huestis MA (2017) Approaches, challenges, and advances in metabolism of new synthetic cannabinoids and identification of optimal urinary marker metabolites. Clin Pharmacol Ther 101:239–253

    Article  CAS  Google Scholar 

  • Diao X, Huestis MA (2019) New synthetic cannabinoids metabolism and strategies to best identify optimal marker metabolites. Front Chem 7:109

    Article  CAS  Google Scholar 

  • European Monitoring Centre for Drugs and Drug Addiction (2017) 5F-MDMB-PINACA—Report on the risk assessment of methyl 2-{[1-(5-fluoropentyl)-1H-indazole-3-carbonyl]amino}-3,3-dimethylbutanoate in the framework of the Council Decision on new psychoactive substances

  • Gamage TF, Farquhar CE, McKinnie RJ et al (2019) Synthetic cannabinoid hydroxypentyl metabolites retain efficacy at human cannabinoid receptors. J Pharmacol Exp Ther 368:414–422

    Article  CAS  Google Scholar 

  • Gatch MB, Forster MJ (2019) Cannabinoid-like effects of five novel carboxamide synthetic cannabinoids. Neurotoxicology 70:72–79

    Article  CAS  Google Scholar 

  • Gurney SMR, Scott KS, Kacinko SL et al (2014) Pharmacology, toxicology, and adverse effects of synthetic cannabinoid drugs. Forensic Sci Rev 26:54–76

    Google Scholar 

  • Harris CR, Brown A (2013) Synthetic cannabinoid intoxication: a case series and review. J Emerg Med 44:360–366

    Article  Google Scholar 

  • Haschimi B, Mogler L, Halter S et al (2019) Detection of the recently emerged synthetic cannabinoid 4F-MDMB-BINACA in ‘legal high’ products and human urine specimens. Drug Test Anal 11:1377–1386

    Article  CAS  Google Scholar 

  • Holm NB, Nielsen LM, Linnet K (2015a) CYP3A4 mediates oxidative metabolism of the synthetic cannabinoid AKB-48. AAPS J 17:1237–1245

    Article  CAS  Google Scholar 

  • Holm NB, Pedersen AJ, Dalsgaard PW, Linnet K (2015b) Metabolites of 5F-AKB-48, a synthetic cannabinoid receptor agonist, identified in human urine and liver microsomal preparations using liquid chromatography high-resolution mass spectrometry. Drug Test Anal 7:199–206

    Article  CAS  Google Scholar 

  • Huang W, Isoherranen N (2018) Development of a dynamic physiologically based mechanistic kidney model to predict renal clearance. CPT Pharmacometrics Syst Pharmacol 7:593–602

    Article  CAS  Google Scholar 

  • Huestis MA, Gorelick DA, Heishman SJ et al (2001) Blockade of effects of smoked marijuana by the CB1-selective cannabinoid receptor antagonist SR141716. Arch Gen Psychiatry 58:322–328

    Article  CAS  Google Scholar 

  • Hussaarts KGAM, Veerman GDM, Jansman FGA et al (2019) Clinically relevant drug interactions with multikinase inhibitors: a review. Ther Adv Med Oncol 11:1–34

    Article  Google Scholar 

  • Izumi S, Nozaki Y, Kusuhara H et al (2018) Relative activity factor (raf)-based scaling of uptake clearance mediated by organic anion transporting polypeptide (OATP) 1B1 and OATP1B3 in human hepatocytes. Mol Pharm 15:2277–2288

    Article  CAS  Google Scholar 

  • Kong TY, Kim JH, Kim DK, Lee HS (2018) Synthetic cannabinoids are substrates and inhibitors of multiple drug-metabolizing enzymes. Arch Pharm Res 41:691–710

    Article  CAS  Google Scholar 

  • Krotulski AJ, Mohr AL, Logan BK (2018) Trend Report: Q4 2018 Synthetic Cannabinoids in the United States

  • Krotulski AJ, Mohr AL, Kacinko SL et al (2019) 4F-MDMB-BINACA: a new synthetic cannabinoid widely implicated in forensic casework. J Forensic Sci 64:1451–1461

    Article  CAS  Google Scholar 

  • Mathialagan S, Piotrowski MA, Tess DA et al (2017) Quantitative prediction of human renal clearance and drug-drug interactions of organic anion transporter substrates using in vitro transport data: a relative activity factor approach. Drug Metab Dispos 45:409–417

    Article  CAS  Google Scholar 

  • Meyer MR, Schütz A, Maurer HH (2015) Contribution of human esterases to the metabolism of selected drugs of abuse. Toxicol Lett 232:159–166

    Article  CAS  Google Scholar 

  • Nguyen HQ, Lin J, Kimoto E et al (2017) Prediction of losartan-active carboxylic acid metabolite exposure following losartan administration using static and physiologically based pharmacokinetic models. J Pharm Sci 106:2758–2770

    Article  CAS  Google Scholar 

  • Noble C, Cannaert A, Linnet K, Stove CP (2019) Application of an activity-based receptor bioassay to investigate the in vitro activity of selected indole- and indazole-3-carboxamide-based synthetic cannabinoids at CB1 and CB2 receptors. Drug Test Anal 11:501–511

    Article  CAS  Google Scholar 

  • Obach RS (1999) Prediction of human clearance of twenty-nine drugs from hepatic microsomal intrinsic clearance data: an examination of in vitro half-life approach and nonspecific binding to microsomes. Drug Metab Dispos 27:1350–1359

    CAS  PubMed  Google Scholar 

  • Poulin P, Theil FP (2002) Prediction of pharmacokinetics prior to in vivo studies. 1. Mechanism-based prediction of volume of distribution. J Pharm Sci 91:129–156

    Article  CAS  Google Scholar 

  • Prakash C, Kamel A, Cui D et al (2000) Identification of the major human liver cytochrome P450 isoform(s) responsible for the formation of the primary metabolites of ziprasidone and prediction of possible drug interactions. Br J Clin Pharmacol 49:35–42

    Article  Google Scholar 

  • Presley BC, Jansen-Varnum SA, Logan BK (2013) Analysis of synthetic cannabinoids in botanical material: a review of analytical methods and findings. Forensic Sci Rev 25:27–46

    CAS  PubMed  Google Scholar 

  • Reidy L, Seither J, Boland D (2018) Identification of synthetic cannabinoid 5-fluoro-ADB in human performance and postmortem samples: a case series. J Forensic Toxicol Pharmacol 7:1–9

    Google Scholar 

  • Riley RJ, McGinnity DF, Austin RP (2005) A unified model for predicting human hepatic, metabolic clearance from in vitro intrinsic clearance data in hepatocytes and microsomes. Drug Metab Dispos 33:1304–1311

    Article  CAS  Google Scholar 

  • Rodgers T, Rowland M (2006) Physiologically based pharmacokinetic modelling 2: predicting the tissue distribution of acids, very weak bases, neutrals and zwitterions. J Pharm Sci 95:1238–1257. https://doi.org/10.1002/jps.20502

    Article  CAS  PubMed  Google Scholar 

  • Seither JZ, Reidy LJ, Boland DM (2020) Identification and quantification of 5-fluoro ADB and the 5-fluoro ADB ester hydrolysis metabolite in postmortem blood samples by LC-MS/MS. J Anal Toxicol 44:133–139

    Article  Google Scholar 

  • Tai S, Fantegrossi WE (2016) Pharmacological and toxicological effects of synthetic cannabinoids and their metabolites. In: Current topics in behavioral neurosciences. Springer Verlag, pp 249–262

  • Thomsen R, Nielsen LM, Holm NB et al (2015) Synthetic cannabimimetic agents metabolized by carboxylesterases. Drug Test Anal 7:565–576

    Article  CAS  Google Scholar 

  • Tran TT, Mittal A, Gales T et al (2004) Exact kinetic analysis of passive transport across a polarized confluent MDCK cell monolayer modeled as a single barrier. J Pharm Sci 93:2108–2123

    Article  CAS  Google Scholar 

  • United Nations Office on Drugs and Crime (2019) Current NPS Threats, vol 1

  • U.S. Food and Drug Administration (2017) Drug Development and Drug Interactions: Table of Substrates, Inhibitors and Inducers. https://www.fda.gov/drugs/drug-interactions-labeling/drug-development-and-drug-interactions-table-substrates-inhibitors-and-inducers. Accessed 18 Aug 2019

  • US FDA (2017) Drug Development and Drug Interactions: Table of Substrates, Inhibitors and Inducers

  • Wang X, Rida N, Shi J et al (2017) A comprehensive functional assessment of carboxylesterase 1 nonsynonymous polymorphisms. Drug Metab Dispos 45:1149–1155

    Article  CAS  Google Scholar 

  • Wang D, Zou L, Jin Q et al (2018) Human carboxylesterases: a comprehensive review. Acta Pharm Sin B 8:699–712

    Article  Google Scholar 

  • Wouters E, Mogler L, Cannaert A et al (2019) Functional evaluation of carboxy metabolites of synthetic cannabinoid receptor agonists featuring scaffolds based on L-valine or L-tert-leucine. Drug Test Anal 11:1183–1191

    Article  CAS  Google Scholar 

  • Yanjiao X, Chengliang Z, Xiping L et al (2013) Evaluation of the inhibitory effects of antihypertensive drugs on human carboxylesterase in vitro. Drug Metab Pharmacokinet 28:468–474

    Article  Google Scholar 

  • Yeter O, Öztürk YE (2019) Detection and quantification of 5F-ADB and its methyl ester hydrolysis metabolite in fatal intoxication cases by liquid chromatography–high resolution mass spectrometry. Forensic Sci Int 302:109866

    Article  CAS  Google Scholar 

  • Yeter O, Ozturk YE (2019) Metabolic profiling of synthetic cannabinoid 5F-ADB by human liver microsome incubations and urine samples using high-resolution mass spectrometry. Drug Test Anal 11:847–858

    Article  CAS  Google Scholar 

  • Zhao A, Tan M, Maung A et al (2015) Rhabdomyolysis and acute kidney injury requiring dialysis as a result of concomitant use of atypical neuroleptics and synthetic cannabinoids. Case Reports Nephrol 2015:1–4

    Article  Google Scholar 

Download references

Acknowledgements

The authors thank Jye Ing Soah, Yen Li Tan and Ching Yee Fong for their assistance in the LC-MS/MS instrumentation. The authors also thank Chi Pang Lui for his feedback. The project is supported by the Singapore Health Sciences Authority (HSA) and the National University of Singapore (NUS) Department of Pharmacy’s Final Year Project (FYP) funding provided to E.C.Y.C. A. C. acknowledges funding as a postdoctoral research fellow from the Research Foundation-Flanders (FWO; 12Y9520N). C. S. acknowledges funding from the Ghent University—Special Research Fund (Grants no. 01N00814 and no. 01J15517).

Author information

Author notes

  1. Wen Lie and Eleanor Jing Yi Cheong have contributed equally as co-first authors.

Authors and Affiliations

  1. Department of Pharmacy, Faculty of Science, National University of Singapore, 18 Science Drive 4, Singapore, 117543, Singapore

    Wen Lie, Eleanor Jing Yi Cheong & Eric Chun Yong Chan

  2. Analytical Toxicology Laboratory, Applied Sciences Group, Health Sciences Authority, 11 Outram Road, Singapore, 169078, Singapore

    Evelyn Mei Ling Goh & Hooi Yan Moy

  3. Laboratory of Toxicology, Department of Bioanalysis, Faculty of Pharmaceutical Sciences, Ghent University, Ottergemsesteenweg 460, 9000, Ghent, Belgium

    Annelies Cannaert & Christophe P. Stove

Authors
  1. Wen Lie
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Eleanor Jing Yi Cheong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Evelyn Mei Ling Goh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hooi Yan Moy
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Annelies Cannaert
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Christophe P. Stove
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Eric Chun Yong Chan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Eric Chun Yong Chan.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher's Note

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

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 254 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lie, W., Cheong, E.J.Y., Goh, E.M.L. et al. Diagnosing intake and rationalizing toxicities associated with 5F-MDMB-PINACA and 4F-MDMB-BINACA abuse. Arch Toxicol 95, 489–508 (2021). https://doi.org/10.1007/s00204-020-02948-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-020-02948-3

Keywords

Search

Navigation