DMD#43505
Supplemental material
Drug Metabolism and Disposition
Identifying a Selective Substrate and Inhibitor Pair for the Evaluation of CYP2J2
Activity
Caroline A. Lee, JP Jones III, Jonathan Katayama, Rüdiger Kaspera, Ying Jiang, Sascha
Freiwald, Evan Smith, Greg Walker, and Rheem A. Totah
Department of Medicinal Chemistry, University of Washington, 1959 Pacific Ave NE,
Seattle, WA 98195. (JPJ, JK, RK, RAT) and Department of Drug Metabolism, Pfizer
Global Research, La Jolla, CA. (CAL, YJ, SF, ES) and Groton, CT(GW)
1
DMD#43505
Tables Legend:
Table1: List of drugs used for to screen for CYP2J2 inhibition.
Figures Legend:
Figure 1: 1H NMR of 4-hydroxy amiodarone compared to amiodarone
Figure 2: 4-HydroxyAmiodarone 1H-1H COSY
Figure 3: 4-HydroxyAmiodarone1H-13C multiplicity edited HSQC
Figure 4: 1H NMR of 3-hydroxy amiodarone compared to amiodarone.
Figure 5: 3-Hydroxy amiodarone 1H-1H COSY
Figure 6: 3-HydroxyAmiodarone 1H-13C multiplicity edited HSQC
Figure 7: Michaelis-Menten Kinetics for 4-OH amiodarone formation with recombinant CYP2J2
2
DMD#43505
Table 1: List of drugs tested for CYP2J2 inhibition
Diphen- Hydrochloro-
Acebutolol Captopril hydramine thiazide Miconazole Probenecid
Diphenyl-
Acetaminophen Carbamazepine hydantoin Ibuprofen Milrinone Procainamide
Acetazolamide Cefazolin Disopyramide Imipramine Minocycline Progesterone
AcycloguanosineChloroquine Dobutamine Indapamide Nabumetone Promazine
Albendazole Chlorpheniramine Domperidone Indomethacin Nadolol Promethazine
Amantadine Chlorpromazine Doxepin Isoniazid Naproxen Propranolol
Amethopterin Chlorthalidone Droperidol Isradipine Nicardipine Quinapril
Amiloride Chlorzoxazone Enalapril Ketoconazole Norfloxacin Quinidine
Amiodarone Cimetidine Erythromycin Ketoprofen Nortriptyline Ramipril
Amitriptyline Cisapride Etodolac Ketorolac Ofloxacin Ranolazine
Amodiaquin Clomipramine Etoposide Labetalol Omeprazole Rifamycin
Amoxicillin Clotrimazole Famotidine Lansoprazole Ondansetron Roxithromycin
Amphotericin Clozapine Felbamate Levofloxacin OrphenadrineSotalol
Antipyrine Colchicine Flecainide Meloxicam Paclitaxel Sparfloxacin
Atenolol Cyclobenzaprine Fluconazole Mesoridazine Pentamidine Spironolactone
Azathioprine CyclophosphamideFluoxetine Methazolamide PentoxifyllineSulindac
Azithromycin Cyclosporin A Flurbiprofen Methimazole Perphenazine Tamoxifen
Benazepril Danazol Fluvoxamine Methotrexate Phenacetin Terfenadine
Bepridil Desipramine Furosemide MethoxypsoralenPimozide Testosterone
3
DMD#43505
Bisoprolol Dexamethasone Gemfibrozil MetoclopramidePiroxicam Thioridazine
Budesonide Diclofenac Glipizide Metolazone Pravastatin Trazodone
Bumetanide Dicloxacillin Griseofulvin Metoprolol Prazosin Vinblastine
Bupropion Digoxin Haloperidol Mevinolin Proadifen Warfarin
4
DMD#43505
Figure 1: 1H NMR of 4-hydroxy amiodarone compared to amiodarone
4‐Hydroxy amiodarone 4 Butyl 1 Butyl 4 Butyl OH 2 Butyl 3 Butyl
8 7 6 5 4 3 2 1 ppm
4 Butyl CH3 4 butyl 3 butyl
2 butyl O 2' I 1 butyl O 6' O 2 DEAE Amiodarone 4 1 DEAE I 1 Butyl 2 Butyl N 4 DEAE 3 Butyl 7 5 H3C 6 CH3 5 DEAE
8 7 6 5 4 3 2 1 ppm
5
DMD#43505
Figure 2: 4-HydroxyAmiodarone 1H-1H COSY
3 Butyl 4 Butyl
1 Butyl 2 Butyl 4 Butyl OH ppm
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 ppm
6
DMD#43505
Figure 3: 4-HydroxyAmiodarone1H-13C multiplicity edited HSQC
4 Butyl OH ppm
10
20 2 Butyl 1 Butyl 3 Butyl 30
40
50
4 Butyl 60
4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 ppm
7
DMD#43505
Figure 4: 1H NMR of 3-hydroxy amiodarone compared to amiodarone.
3‐HydroxyAmiodarone 4 Butyl
1 3 Butyl OH Butyl 3 Butyl 2 Butyl
8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 ppm CH3 4 butyl 3 butyl 4 Butyl 2 butyl O 2' I 1 butyl O 6' O 2 DEAE 4 I 1 DEAE N 4 DEAE 1 Butyl 7 5 H3C 2 Butyl 3 Butyl 6 CH3 5 DEAE
Amiodarone
8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 ppm
8
DMD#43505
Figure 5: 3-Hydroxy amiodarone 1H-1H COSY
1 Butyl 4 Butyl 3 Butyl
4 Butyl OH 2 Butyl ppm
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 ppm
9
DMD#43505
Figure 6: 3-HydroxyAmiodarone 1H-13C multiplicity edited HSQC
4 Butyl OH ppm
1 Butyl weak 10
4 Butyl 20
30
2 Butyl 40
50
60 3 Butyl
70
4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 ppm
10
DMD#43505
Figure 7: Michaelis-Menten Kinetics for 4-OH amiodarone formation with recombinant CYP2J2
4
3
Vmax = 4.6 pmol/min/pmol CYP2J2 2 Km = 5.0 M
1 4-Hydroxyamiodarone (pmol/min/pmol CYP2J2) (pmol/min/pmol 0 0 10 20 30 40 50 60 Amiodarone (M)
11