AM-2201 Inhibits Multiple Cytochrome P450 and Uridine 5′-Diphospho-Glucuronosyltransferase Enzyme Activities in Human Liver Microsomes

Abstract

AM-2201 is a synthetic cannabinoid that acts as a potent agonist at cannabinoid receptors and its abuse has increased. However, there are no reports of the inhibitory effect of AM-2201 on human cytochrome P450 (CYP) or uridine 5′-diphospho-glucuronosyltransferase (UGT) enzymes. We evaluated the inhibitory effect of AM-2201 on the activities of eight major human CYPs (1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, and 3A4) and six major human UGTs (1A1, 1A3, 1A4, 1A6, 1A9, and 2B7) enzymes in pooled human liver microsomes using liquid chromatography–tandem mass spectrometry to investigate drug interaction potentials of AM-2201. AM-2201 potently inhibited CYP2C9-catalyzed diclofenac 4′-hydroxylation, CYP3A4-catalyzed midazolam 1′-hydroxylation, UGT1A3-catalyzed chenodeoxycholic acid 24-acyl-glucuronidation, and UGT2B7-catalyzed naloxone 3-glucuronidation with IC50 values of 3.9, 4.0, 4.3, and 10.0 µM, respectively, and showed mechanism-based inhibition of CYP2C8-catalyzed amodiaquine N-deethylation with a Ki value of 2.1 µM. It negligibly inhibited CYP1A2, CYP2A6, CYP2B6, CYP2C19, CYP2D6, UGT1A1, UGT1A4, UGT1A6, and UGT1A9 activities at 50 μM in human liver microsomes. These in vitro results indicate that AM-2201 needs to be examined for potential pharmacokinetic drug interactions in vivo due to its potent inhibition of CYP2C8, CYP2C9, CYP3A4, UGT1A3, and UGT2B7 enzyme activities.

In Vitro Interaction of AB-FUBINACA with Human Cytochrome P450, UDP-Glucuronosyltransferase Enzymes and Drug Transporters

“…AB-FUBINACA competitively inhibited CYP2C8-catalyzed amodiaquine N -de-ethylation and CYP2C9-mediated diclofenac 4′-hydroxylation with K i values of 19.9 and 13.1 μM, respectively: other synthetic cannabinoids including AM-2201, EAM-2201, and MAM-2201 potently inhibited CYP2C8-catalyzed amodiaquine N -de-ethylation ( K i , 0.54–2.1 µM) and CYP2C9-mediated diclofenac 4′-hydroxylation ( K i , 3.0–5.6 µM) by ultrapooled human liver microsomes [ 27 , 28 , 29 ]. AB-FUBINACA exhibited mixed inhibition of CYP2B6-mediated bupropion hydroxylation with a K i of 15.0 μM.…”

“…AM-2201 ( K i , 4.0 µM), MAM-2201 ( K i , 5.4 µM), EAM-2201 ( K i , 4.1 μM and k inact , 0.0250 min −1 ), and APINACA ( K i , 4.5 µM; k inact , 0.04686 min −1 ) inhibited CYP3A4-catalyzed midazolam 1′-hydroxylation in human liver microsomes [ 27 , 28 , 29 , 30 ], but AB-FUBINACA did not inhibit the CYP3A4, CYP1A2, and CYP2A6 activities of such microsomes ( Figure 2 ).…”

“…AB-FUBINACA negligibly inhibited the UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A9, and UGT2B7 activities of human liver microsomes, suggesting that the risk of clinical DDIs between AB-FUBINACA and the UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A9, and UGT2B7 metabolizing enzymes is remote. However, EAM-2201, MAM-2201, and AM-2201 competitively inhibited UGT1A3-mediated chenodeoxycholic acid 24-acyl-β-glucuronidation with K i values of 2.7, 4.3, and 5.0 μM, respectively, and APINACA noncompetitively inhibited UGT1A9-mediated mycophenolic acid glucuronidation ( K i , 5.9 µM) by human liver microsomes [ 27 , 28 , 29 , 30 ].…”

“…The inhibitory potentials (IC 50 values) of AB-FUBINACA on CYP 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, and 3A4 activities in pooled human liver microsomes were evaluated using a slight modification of our previously described method that employed a cocktail of CYP substrates, followed by LC-MS/MS [ 27 , 28 , 29 , 30 ]. The incubation mixtures were prepared in total volumes of 100 μL as follows: 50 mM potassium phosphate buffer (pH 7.4), 1.0 mM NADPH, 10 mM MgCl 2 , ultrapooled human liver microsomes (0.2 mg/mL), various concentrations of AB-FUBINACA (final concentrations of 0.1–100 μM), and the CYP enzyme-specific substrates of the cocktail sets A (2.0 μM amodiaquine, 5 μM bufuralol, 2.5 μM coumarin, 10 μM diclofenac, 100 μM [ S ]-mephenytoin, 2.5 μM midazolam, and 50 μM phenacetin) and B (50 μM bupropion).…”

“…Drug abusers frequently take several drugs concomitantly [ 13 , 14 , 15 ]; it is thus necessary to investigate the inhibitory effects of drugs on major drug-metabolizing enzymes such as cytochrome P450s (CYPs) and 5′-diphospho-glucuronosyltransferases (UGTs); and on clinically important drug transporters such as solute carrier transporters including the organic cation transporters (OCT)1, and OCT2, the organic anion transporters (OAT)1 and OAT3, the organic anion transporting polypeptides (OATP)1B1 and OATP1B3, and efflux transporters such as P-glycoprotein (P-gp) and breast cancer-resistance protein (BCRP) [ 16 , 17 , 18 ]. The inhibitory effects of phytocannabinoids including THC, cannabinol, cannabidiol [ 19 , 20 , 21 , 22 , 23 , 24 , 25 ], and synthetic cannabinoids (JWH-019, STS-135, UR-144, AM-2201, MAM-2201, EAM-2201, and APINACA) [ 4 , 26 , 27 , 28 , 29 , 30 ], on the major CYP and UGT enzymes or human liver microsomes, or recombinant CYP and UGT enzymes, have been reported [ 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 ].…”