Screening of Specific Inhibitors for Human Carboxylesterases or Arylacetamide

Deacetylase

Mai Shimizu, Tatsuki Fukami, Miki Nakajima, and Tsuyoshi Yokoi

Drug Metab Dispos

Supplemental Table 1. Chemicals used in this study. Alfa Aesar (Tokyo, Japan) 3-(4-Fluorobenzoyl) propiponic Acid, 3-Methylflavone-8-carboxylic acid

BD Gentest (Woburn, MA) a25-Hydroxycholesterol, b4'-Hydroxydiclofenac, b5-Hydroxydiclofenac

Biomol international (Plymouth Meeting, PA) Pizotifen malate

Merk (Darmstadt, Germany) Mevastatin, Resveratrol

Carbosynthe (Berkshire, UK) N-Acetyl-l-cysteine, Adefovir, 5,6-Dichlorobenzimidazole 1-β-d-Riboburanoside, Indiplon, Milnacipran hydrochloride, Pyridoxal 5'-phosphate

Cayman Chemical (Ann Arbor, MI) N-(4-Hydroxyphenyl)-arachidonoyl amide, JW480

Dojindo Laboratories (Kumamoto, Japan) N-Cyclohexyl-3-aminopropanesulfonic acid, 3,3'-Diaminobenzidine tetrahydrochloride, Ethylene glycol-bis (β-aminoethylether)-N,N,N',N'-tetraacetic acid

Enzo Life Science (Farmingdale, NY) Castanospermine, Pregnenolone-16α-carbonitrate

Fujicco (Kobe, Japan) Daidzein

Funakoshi (Tokyo, Japan) Blasticidin S hydrochloride, Chloroquine, Coumarin, Diethyldithiocarbamate, 7-Ethoxy-4-(trifluoromethyl) coumarin, Furafylline, G-418 sulfate, Ketoconazole, MelQ, l-α-Phosphatidyl choline dilauroyl, Potassium ferricyanide, SKF-525A, Tetrabutylammonium hydrogen sulfate, WST-1

Life Technologies (Catlsbad, CA) Cadmium Chloride

LKT Laboratories (St. Paul, MN) Bestatin hydrochloride, Capecitabine, Clotrimazole, Doxifluridine, Enalaprilat, Ezetimibe, Ftorafur, Indole-3-carbinol, Montelukast, Rebamipide, Rosiglitazone, Tegafur, bTelmisartan, Temocapril hydrochloride, Terazosin hydrochloride, Tolmetin sodium, Toremifene base

MP Biochemicals (Tokyo, Japan) Arachidonic acid

Nacalai Tesque (Kyoto, Japan) Sodium laurylsulfate

Research Organics (Cleveland, OH) N-Acetyl procainamide hydrochloride

Santa Cruz Biotechnology (Santa Cruz, CA) Benzyl-2-acetamido-2-deoxy-α-d-galactopyranoside, Camostat mesylate, (+)-Clopidogrel hydrogen sulfate, Dicloxacillin sodium salt monohydrate, d-Mannose-6-phosphate disodium salt, Metyrapon, 1-Naphthyl acetate, α-Naphthoflavone, Ondansetron hydrochloride, Perphenazine, Sparteine, Sulfaphenazole, Thiouric acid sodium salt dehydrate, Tryptamine hydrochloride, WY-14643

Sigma-Aldrich Acebutolol hydrochloride, Acemetacin, p-Acetamidobenzoic acid, 9-Acetylanthracene, Acetyl-dl-carnitine hydrochloride, Amantadine hydrochloride, p-Aminobenzoic acid, 2-Aminobenzylalcohol, 6-Aminochrysene, 2-Aminofluorene, Amiodarone hydrochloride, Ammonium pyrrolidinedithiocarbamate, 5α-Androstane-3α,17β-diol, 2-Anthramine, Astaxanthin, 5-Azacytidine, Barbital, Bezafibrate, Benzbromarone, 6-Benzyl-2-thiouracil, 5-Bromouracil, Bupropion, Busulfan, l-Buthionine-[S,R]-sulfoximine, Caffeine, Cefamandole, Cefoperazone, 2-Chloro-3',4'-dimethoxybenzil, 5-Chlorouracil, Chlorpropamide, Chlorzoxazone, Chrysin, Clofibric acid, Cloxacillin sodium salt monohydrate, (-)-Cotinine, Cyclobenzaprine hydrochloride, Dapson, Deferoxamine mesylate salt, Dehydroisoandrosterone-3-sulfate, Deoxycorticosterone, Diazepam, Diclofenac sodium salt, (-)-3-(3,4-Dihydroxyphenyl)-2-methyl-l-alanine sesquihydrate, 2,6-Dihydroxypyridine hydrochloride, Dimethyl pimelimidate dihydrochloride, Diosmin, Doxepin hydrochloride, 17-Epiestriol, (-)-Epinephrine (+)-bitartratee, β-Estradiol, Estriol, Estrone, Estrone 3-sulfate potassium salt, dl-Ethionine, 7-Ethoxycoumarin, Ethoxyresorufin, 5-Ethyl-5-p-tolybarbituric acid, Fenbendazole, Fenofibrate, Ficoll 400, Flunitrazepam, 5-Fluoro-2'-deoxyuridine, 5-Fluorouridine, Fluoxetine, Flurbiprofen, Gadolinium (III) chloride hexahydrate, Hecogenin acetate, Hexobarbital, Hydralazine hydrochloride, 3-Hydroxybenzylhydrazine dihydrochloride, 7-Hydroxycoumarin, 12-Hydroxydodecanoic acid, 11β-Hydroxytestosterone, 3-Hydroxybenzylhydrazine dihydrochloride 2-Hydroxy-1,4-naphthoquinone, 5-(3-Hydroxyphenyl)-5-phenylhydantoin, 5-(4-Hydroxyphenyl)-5-phenylhydantoin, 17α-Hydroxyprogesterone, 7-(β-Hydroxypropyl) theophylline, dl-Isocitric acid, Isoniazid, Kaempferol, Labetalol hydrochloride, Lauric acid sodium salt, Z-Leu-Leu-Leu-al, Leupeptin, Levofloxacin, Loxapine succinate salt, Maprotiline hydrochloride, Methapyrilene hydrochloride, Methimazole, (S)-(+)-6-Methoxy-α-methyl-2-naphthaleneacetic acid, 3-Methylindole, 4-Methylumbelliferone, 5-(4-Methylphenyl)-5-phenyl-hydantoin, 1-Methyl-4-phenyl-1,2,3,6-tetrahydro-pyridine hydrochloride, Nalidixic acid, Naloxone hydrochloride, β-Naphthoflavone, 1,2-Naphthoquinone, Nialamide, (-)-Nicotine, Nicotinic acid N-oxide, Nimesulide, Nimodipine, Nordihydroguaiaretic acid, l-(-)-Norepinephrine, Norfloxacin, Nortriptyline hydrochloride, Omeprazole, Oxibendazole, Oxybutiynin chloride, Paclitaxel, Pepstatin A, 1,7-Phenanthroline, Phenazine ethosulfate, p-Phenetidine, (R)-1-Phenyl-1,2-ethanediol, (S)-1-Phenyl-1,2-ethanediol, Pilocarpine hydrochloride, Polymyxin B sulfate salt, Praziquantel, Prednisolone, 5α-Pregnan-3,20-dione, 5-Pregnen-3β-ol-20-one-16α-carbonitrile, Prilocaine hydrochloride, Procainamide hydrochloride, Procaine hydrochloride, Prochlorperazine dimaleate salt, Progesterone, Proglumide, Promazine hydrochloride, (R)-(+) Propranolol hydrochloride, 6-Propyl-2-thiouracil, Protriptyline hydrochloride, Pyrazole, Quercetin, d-Saccharic acid 1,4-lactone, Sodium orthovanadate, Sulfamethazine, Tetrabutylammonium hydroxide, Thiobenzamide, l-Thyroxine, (±)-α-Tocopherol nicotinate, Triacetyloleandomycin, Trichostatin A, Trimipramine maleate, l-Tryptophan, Uracil, Uridine, Uridine 5'-diphosphate sodium salt, R,S-Warfarin, Zolpidem, Zomepirac sodium salt

Steraloids (Newport, RI) 5β-Pregnan-3,20-dione

Tokyo Chemical Industry (Tokyo, Japan) 1-Aminobenzotriazole, 2-Amino-m-cresol, 4-Amino-3,5-xylenol, Benzo[a]anthracene, Captopril, Dehydroepiandrosterone, Delapril hydrochloride, 2,2'-Dihydroxy-4-methoxybenzophenone, Dimethyl pimelimidate dihydrochrolide, 4-Hydroxybenzoic acid n-hexyl ester, Methotrexate, Methylviologen, Mifepristone, 1-(1-Naphthyl)-2-thiourea, Oxymetholine, Spironolactone, 1,2,3,6-Tetrahydro-4-phenylpyridine hydrochloride

Toronto Research Chemicals N-Acetyl dapsone, Albendazole sulfoxide, Azasetron, Bufuralol hydrochloride, Butenafin hydrochloride, Candesartan, Cefmetazole, 5-Chloro-2,4-dihydroxypyridine, Digoxin, Docetaxel, Eprosartan, Fenofibric acid, GW4064, 6-Hydroxychlorzoxazone, 3-Hydroxy desloratadine, (+/-)-4'-Hydroxy mephenytoin, Imidapril hydrochloride, Irbesartan, Irinotecan, Isocarboxazid, Losartan carboxaldehyde, Losartan carboxylic acid, Mephenytoin, Nateglinide, Olmesartan, Oxonic acid potassium salt, Pitavastatin lactone, Prasugrel hydrochloride, Quetiapine hemifumarate, Raloxifene hydrochloride, Sertraline hydrochloride, Simvastatin, SN-38, Tolcapone, 3,3',5-Triiodo-l-thyronine, Valsartan, Vanillylamine hydrochloride

Ultrafine Chemicals and Minerals (Tamilnadu, India) Debrisoquine sulphate, Dextromethorphan hydrobromide, Oxydized nifedipine, Terfenadine

Wako Pure Chemical Industries Acarbose, Aceclofenac, Acetanilide, Acetoaminophen, Acetohexamide, 7-Acetoxy-4-methylcoumarin, Acetylsalicylic acid, Albiflorin, Allopurinol, (p-Amidinophenyl) methanesulfonyl fluoride hydrochloride, dl-2-Aminobutyric acid, Aminoguanidine hydrochloride, 6-Aminohexanoic acid, 5-Aminolevulinic acid hydrochloride, 1-Amino-2-methylindoline, 2-Amino-5-nitrobenzophenone, 5-Amino-2-nitrobenztrifluoride, 3-Aminophenol, 3-Amino-1H-1,2,4-triazole, 4-Amino-1,2,4-triazole, 4-Amino-2-(trifluoromethyl)benzonitrile, Amitriptyline hydrochloride, Amoxapine, Ampicillin sodium, (+)-4-Androstene-3,17-dione, l(+)-Arabinose, 5-Aza-2'-deoxycitidine, 3'-Azido-3'-deoxythymidine, Baicalein, Baicalin, Barium hydroxide octahydrate, p-Benzoyl phenol, Benzhydrol, Benzophenone, Benzylhydrazine dihydrochloride, Bicalutamide, Bilirubin, Borneol, Buformine hydrochloride, t-Butylhydroquinone, S-Butyrylthiocholine iodide, Calcitriol, Camptothecin, Capsaicin, Carbamazepine, Carbomethoxymethylene, Carvedilol, Cefazolin sodium salt, Chenodeoxycholic acid, 3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate, Chloramphenicol, p-Chlorobenzoic acid, 4-Chloro-1-naphthol, Chlorpromazine hydrochloride, Ciclosporine, Ciprofloxacin hydrochloride monohydrate, Clarithromycin, Clomipramine hydrochloride, Clopidogrel carboxylic acid, Clozapine, Colchicine, Corticosterone, Coumarin-3-carboxylic acid, o-Cresol, p-Cresol, Cyclizine hydrochrolide, Cycloheximide, 2-Cyclohexyl-2-phenylglycolic Acid, Cyclophosphamide monohydrate, l-Cysteine, Cytidine, Cytosine-1-β-d(+)-arabinofuranoside, Dantrolene sodium, 1-Decanesulfonic acid, Desipramine hydrochloride, Dexamethasone, cis-Diammine dichloroplatinum (II), Dibucaine hydrochloride, 2,6-Di-tert-butyl-4-methylphenol, N6,2'-O-Dibutyryladenosine 3',5'-(cyclic)monophosphate sodium salt, 2,6-Dichloro-4-nitrophenol, 3,4-Dichloroaniline, 3,4-Dichloronitrobenzene, 2,6-Dichlorophenol, Digitonin, 1,2-Dihydroxy-4-nitrobenzene, dl-β-(3,4-Dihydroxyphenyl) alanine, Diltiazem hydrochloride, (±)-Dithiothreitol, p-Dimethylaminobenzaldehyde, 1,3-Dimethyluric acid, Diphenhydramine hydrochloride, Disodium molybdate (VI) dehydrate, Disulfiram, Dopamine hydrochloride, Enalapril maleate, Erythromycin, Ethacrynic acid, Ethylcarbamate, 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, N-Ethylmaleimide, Ethynylestradiol, Etizolam, Etodolac, Etoposide, Famotidine, Felbamate, Flavoxate hydrochloride, Fluconazole, Fluoranthene, p-Fluorophenyl acetic acid, 5-Fluorouracil, Flutamide, Fluvastatin sodium, o-Formylbenzoic acid, Fulvestrant, 2-Furancarboxylic, Furosemide, d-Galactosamine hydrochloride, Gemfibrozil, Genistein, Gentamicin sulfate, Glibenclamide, Gliclazide, l-Glutamic acid, Glycitein, Glycyrrhizic acid dipotassium salt, Guanidine hydrochloride, Haloperidol, Harmin hydrochloride, 1-Heptanesulfonic acid sodium salt, Hexaammonium heptamolybdate tetrahydrate, 1-Hexanesulfonic acid sodium salt, Histamine dihydrochloride, l-Histidine, Hydroquinone, o-Hydroxybenzamide, 16α-Hydroxyprogesterone, Ibuprofen, Imidazole, Imipramine hydrochloride, Indapamide, Iodoacetamide, 3-Indoleacetic acid, 3-Indoleethanol, Indomethacin, Iproniazid phosphate, Isopropyl-β-d(-)-thiogalactopyranoside, Ketoprofen, Ketotifen fumarate, Leflunomide, Lithocholic acid, Loperamide hydrochloride, Lorazepam, Losartan potassium, Lovastatin, Medroxyprogesterone acetate, Melatonin, l(-)-Menthol, Metformin, 5-Methoxy-2-methylindole-3-acetic acid, 5-Methoxytryptamine hydrochloride, 3-Methylxanthine, Metoprolol tartrate, Monobromobimane, Monomethylol dimethylhydantoin, Mycophenolic acid, 1-Naphthol, 2-Naphthol, 1,4-Naphthoquinone, 1-Naphthyl isothiocyanate, Nefazodone hydrochloride, Nickel (II) sulfate hexahydrate, Niflumic acid, Nilutamide, 2-Nitrobenzoic acid, 1-Octanesulfonic acid sodium salt, Oxazepam, Oxcarbazepine, Paeoniflorin, Palmitic acid, Papaverine, 1-Pentanesulfonic acid sodium salt, Phenacetin, l-(-)-Phenylalanine, trans-2-Phenylcyclopropylamine hydrochloride, Phenylbutazone, Phenylmethylsulfonyl fluoride, Phenytoin, PhIP hydrochloride, Phthalaldehyde, Pioglitazone, Potassium 4-nitrophenyl sulfate, Pravastatin sodium salt, Prazosin hydrochloride, Probenecid, Probucol, Propanil, dl-Propranolol hydrochloride, Pyrazinamide, Pyrazinecarboxylic acid, Pyrene, Pyrimethamine, Quinidine sulfate dehydrate, Quinine, Quinine sulfate dehydrate, 8-Quinolinol, Ranitidin hydrochloride, Rosuvastatin, Roxithromycin, Rubidium chloride, Semicarbazide hydrochloride, Sennidine A, Sennoside A, Sodium hydrogen sulfite, Sodium N-lauroyl salcocinate, Sodium taurocholate, Sodium tungstate (VI) dehydrate, Spermine, Strontium chloride hexahydrate, Succinic acid, Sulfamethoxazole, Sulfasalazine, Sulfobromophthalein sodium salt, 5-Sulfosalicylic acid dehydrate, Sulindac, (±)-Sulpiride, Tacrine hydrochloride, Tamoxifen citrate, Teprenone, Terbinafine, Terbutaline, Testosterone, Tetracycline, Tetramethyl ammonium chloride, Theobromine, Theophylline, Thiamine hydrochloride, Thioacetamide, Thiourea, Thymidine, Thymine, Ticlopidine hydrochloride, Tolbutamide, Tolperisone hydrochloride, Tranilast, Trazodone hydrochloride, Tricine, Troglitazone, Trp-P-1 acetate, Tryptamine hydrochloride, l-Tyrosine, U0126, Valproic acid sodium salt, Verapamil hydrochloride, Vinblastine sulfate, Wogonin standard, Zafirlukast a The compound was dissolved in ethanol. b The compounds were dissolved in methanol. The other compounds were dissolved in DMSO.

Supplemental Table 2. Inhibitory effects of 100 µM various compounds on PNPA hydrolase activities. % of control Compounds CES1 CES2 AADAC Acarbose 85 95 101 Acebutolol hydrochloride 80 86 101 Aceclofenac 17 43 37 Acemetacin 0 0 0 p-Acetamidobenzoic acid 86 90 100 Acetanilide 79 60 64 Acethexamide 84 52 105 Acetoaminophen 90 100 90 7-Acetoxy-4-methylcoumarin 76 45 26 9-Acetylanthracene 29 73 55 Acetyl-dl-carnitine hydrochloride 78 103 93

N-Acetyl-L-cysteine 87 86 85 N-Acetyl dapsone 81 48 97 N-Acetyl procainamide hydrochloride 93 89 100 Acetylsalicylic acid 66 86 92 Adefovir 36 9 52 Albendazole sulfoxide 98 89 102 Albiflorin 91 96 97 Allopurinol 74 120 105 Amantadine hydrochloride 74 97 142 (p-Amidinophenyl) methanesulfonyl fluoride hydrochloride 64 79 82 p-Aminobenzoic acid 73 94 95 1-Aminobenzotriazole 86 114 93 2-Aminobenzylalcohol 81 107 108

DL-2-Aminobutyric acid 89 96 65 6-Aminochrysene 43 11 77 2-Amino-m-cresol 93 90 86 2-Aminofluorene 52 68 100 Aminoguanidine hydrochloride 89 98 103 6-Aminohexanoic acid 105 99 65 5-Aminolevulinic acid hydrochloride 97 106 103 1-Amino-2-methylindoline 67 24 58 2-Amino-5-nitrobenzophenone 97 90 87 5-Amino-2-nitrobenztrifluoride 63 108 117 3-Aminophenol 85 113 92 3-Amino-1H-1,2,4-triazole 70 98 94 4-Amino-1,2,4-triazole 104 100 54 4-Amino-2-(trifluoromethyl)benzonitrile 34 65 92 4-Amino-3,5-xylenol 96 87 87 Amiodarone hydrochloride 65 30 93 Amitriptyline hydrochloride 42 28 89 Ammonium pyrrolidinedithiocarbamate 74 92 87 Amoxapine 34 45 75 Ampicillin sodium 95 98 95 5α-Androstane-3α,17β-diol 9 45 87 (+)-4-Androstene-3,17-dione 57 111 112 2-Anthramine 82 26 106

L(+)-Arabinose 89 96 99 Arachidonic acid 19 23 41 Astaxanthin 76 47 65 5-Azacytidine 69 81 79 5-Aza-2'-deoxycitidine 77 70 84 Azasetron 99 90 88 3'-Azido-3'-deoxythymidine 66 100 83 Baicalein 19 42 22 Baicalin 106 95 91 Barbital 83 106 98 Barium hydroxide octahydrate 84 70 89 Benzbromarone 6 4 3 Benzhydrol 27 35 96 Benzo[a]anthracene 80 72 105 Benzophenone 17 46 141 p-Benzoyl phenol 5 29 80

Benzyl-2-acetamido-2-deoxy-α-D-galactopyranoside 104 88 105 Benzylhydrazine dihydrochloride 83 52 98 6-Benzyl-2-thiouracil 131 88 100 Bestatin hydrochloride 111 103 102 Bezafibrate 107 70 78 Bicalutamide 43 58 42 Bilirubin 350 145 135 Blasticidin S hydrochloride 92 82 100 Borneol 35 48 75 5-Bromouracil 77 94 82 Buformine hydrochloride 87 85 92 Bufuralol hydrochloride 79 53 105 Bupropion 9 45 40 Busulfan 76 80 82 Butenafin hydrochloride 65 13 100

L-Buthionine-[S,R]-sulfoximine 53 97 65 t-Butylhydroquinone 47 50 66 S-Butyrylthiocholine iodide 95 87 91 Cadmium chloride 104 103 101 Caffeine 92 93 84 Calcitriol 5 25 53 Camostat mesylate 81 70 80 Camptothecin 91 87 79 Candesartan 73 40 20 Capecitabine 91 105 103 Capsaicin 12 12 45 Captopril 100 119 107 Carbamazepine 68 112 68 Carbomethoxymethylene 73 156 96 Carvedilol 5 3 24 Castanospermine 104 98 79 Cefamandole 97 83 94 Cefazolin sodium salt 78 75 91 Cefmetazole 87 69 76 Cefoperazone 86 78 88 Chenodeoxycholic acid 9 67 13 Chloramphenicol 96 94 92 2-Chloro-3',4'-dimethoxybenzil 7 13 106 p-Chlorobenzoic acid 85 86 99 5-Chloro-2,4-dihydroxypyridine 104 140 122 4-Chloro-1-naphtol 7 6 15 Chloroquine 95 100 79 5-Chlorouracil 101 136 93 Chlorpromazine hydrochloride 15 16 45 Chlorpropamide 94 124 92 Chlorzoxazone 77 77 98 3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate 99 121 121 Chrysin 84 194 109 Ciclosporine 126 75 96 Ciprofloxacin hydrochloride monohydrate 114 95 99 Clarithromycin 77 101 62 Clofibric acid 87 78 91 Clomipramine hydrochloride 15 30 61 Clopidogrel carboxylic acid 70 69 87 (+)-Clopidogrel hydrogen sulfate 10 28 83 Clotrimazole 70 59 29 Cloxacillin sodium salt monohydrate 101 118 116 Clozapine 44 7 60 Colchicine 94 86 70 Corticosterone 32 166 55 (-)-Cotinine 115 111 104 Coumarin 93 98 78 Coumarin-3-carboxylic acid 92 115 117 o-Cresol 89 74 83 p-Cresol 77 86 82 Cyclizine hydrochrolide 58 68 77 Cyclobenzaprine hydrochloride 21 28 67 Cycloheximide 92 74 85 N-Cyclohexyl-3-aminopropanesulfonic acid 101 133 169 2-Cyclohexyl-2-phenylglycolic acid 79 83 94 Cyclophosphamide monohydrate 95 108 92 L-Cysteine 91 130 81 Cytidine 85 107 86

Cytosine-1-β-D(+)-arabinofuranoside 76 115 100 Daidzein 24 84 57 Dantrolene sodium 82 47 104 Dapson 32 80 97 Debrisoquine sulphate 91 95 105 1-Decanesulfonic acid 61 230 119 Deferoxamine mesylate salt 84 80 87 Dehydroepiandrosterone 23 75 57 Dehydroisoandrosterone-3-sulfate 42 43 118 Delapril hydrochloride 84 65 110 Deoxycorticosterone 37 87 90 Desipramine hydrochloride 24 60 72 Dexamethasone 78 71 83 Dextromethorphan hydrobromide 69 45 98 3,3'-Diaminobenzidine tetrahydrochloride 53 45 78 cis-Diammine dichloroplatinum (II) 49 29 72 Diazepam 28 12 91 Dibucaine hydrochloride 38 23 107 2,6-Di-tert-butyl-4-methylphenol 38 34 164 N6,2'-O-Dibutyryladenosine 3',5'-(cyclic)monophosphate sodium salt 104 89 91 3,4-Dichloroaniline 92 96 91 5,6-Dichlorobenzimidazole 1-β-D-riboburanoside 94 101 98 3,4-Dichloronitrobenzene 66 96 97 2,6-Dichloro-4-nitrophenol 0 0 0 2,6-Dichlorophenol 8 20 44 Diclofenac sodium salt 10 20 72 Dicloxacillin sodium salt monohydrate 69 88 85 Diethyldithiocarbamate 12 34 77 Digitonin 14 87 96 Digoxin 92 16 89 2,2'-Dihydroxy-4-methoxybenzophenone 6 7 71 1,2-Dihydroxy-4-nitrobenzene 66 121 56

DL-β-(3,4-Dihydroxyphenyl) alanine 92 106 90 (-)-3-(3,4-Dihydroxyphenyl)-2-methyl-L-alanine sesquihydrate 98 96 98 2,6-Dihydroxypyridine hydrochloride 82 82 98 Diltiazem hydrochloride 19 41 86 p-Dimethylaminobenzaldehyde 67 102 106 Dimethyl pimelimidate dihydrochrolide 82 93 92 1,3-Dimethyluric acid 92 101 115 Diosmin 79 59 74 Diphenhydramine hydrochloride 42 52 86 Disodium molybdate (VI) dihydrate 91 100 82 Disulfiram 10 35 108 (±)-Dithiothreitol 92 104 105 Docetaxel 70 44 60 Dopamine hydrochloride 67 50 83 Doxepin hydrochloride 37 31 86 Doxifluridine 48 10 93 Enalapril maleate 96 103 97 Enalaprilat 100 107 99 17-Epiestriol 11 24 31 (-)-Epinephrine (+)-bitartratee 124 102 96 Eprosartan 57 20 69 Erythromycin 55 78 94 β-Estradiol 17 65 50 Estriol 8 46 47 Estrone 56 64 75 Estrone 3-sulfate potassium salt 82 88 106 Ethacrynic acid 70 104 72

DL-Ethionine 102 110 105 7-Ethoxycoumarin 57 57 75 Ethoxyresorufin 94 77 109 7-Ethoxy-4-(trifluoromethyl) coumarin 47 61 90 Ethylcarbamate 102 104 90 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride 94 98 84 Ethylene glycol-bis (β-aminoethylether)-N,N,N',N''-tetraacetic acid 71 81 72 N-Ethylmaleimide 94 88 93 5-Ethyl-5-p-tolybarbituric acid 70 101 106 Ethynylestradiol 0 18 20 Etizolam (50 µM) 85 71 113 Etodolac 66 78 69 Etoposide 110 79 95 Ezetimibe 16 6 41 Famotidine 103 97 79 Felbamate 93 102 88 Fenbendazole 50 52 94 Fenofibrate 58 5 71 Fenofibric acid 75 60 102 Ficoll 400 111 104 106 Flavoxate hydrochloride 85 10 107 Fluconazole (50 µM) 106 91 113 Flunitrazepam 52 58 113 Fluoranthene 57 56 100 3-(4-Fluorobenzoyl) propiponic acid 83 93 86 5-Fluoro-2'-deoxyuridine 76 82 83 p-Fluorophenyl acetic acid 85 82 87 5-Fluorouracil 103 88 93 5-Fluorouridine 92 96 97 Fluoxetine 21 37 68 Flurbiprofen 43 107 70 Flutamide 33 39 113 Fluvastatin sodium 69 55 64 Fluvestrant 51 8 66 o-Formylbenzoic acid 102 103 97 Ftorafur 76 85 88 Furafylline 92 109 97 2-Furancarboxylic 80 81 94 Furosemide 82 79 96 Gadolinium (III) chloride hexahydrate 81 87 88

D-Galactosamine hydrochloride 74 83 90 Gemfibrozil 66 102 120 Genistein 24 56 30 Gentamicin sulfate 106 113 88 Glibenclamide 20 4 67 Gliclazide 102 96 98

L-Glutamic acid 90 90 90 Glycitein 51 80 71 Glycyrrhizic acid dipotassium salt 93 99 110 G-418 sulfate 75 74 103 Guanidine hydrochloride 91 94 85 GW4064 4 13 52 Haloperidol 101 29 88 Harmin hydrochloride 51 54 91 Hecogenin acetate 119 92 89 1-Heptanesulfonic acid sodium salt 95 95 108 1-Hexanesulfonic acid sodium salt 103 103 92 Hexaammonium heptamolybdate tetrahydrate 95 94 97 Hexobarbital 82 83 84 Histamine dihydrochloride 106 102 85

L-Histidine 115 98 91 Hydralazine hydrochloride 97 75 93 Hydroquinone 103 93 92 o-Hydroxybenzamide 89 81 84 4-Hydroxybenzoic acid n-hexyl ester 10 11 12 3-Hydroxybenzylhydrazine dihydrochloride 80 61 40 6-Hydroxychlorzoxazone 94 59 104 25-Hydroxycholesterol 35 46 89 7-Hydroxycoumarin 73 56 63 3-Hydroxy desloratadine 45 41 51 4'-Hydroxydiclofenac 8 7 13 5-Hydroxydiclofenac 25 26 14 12-Hydroxydodecanoic acid 79 57 118 2-Hydroxy-1,4-naphthoquinone 96 70 109 (+/-)-4'-Hydroxy mephenytoin 86 66 99 N-(4-Hydroxyphenyl)-arachidonoyl amide 32 10 82 5-(3-Hydroxyphenyl)-5-phenylhydantoin 82 60 74 5-(4-Hydroxyphenyl)-5-phenylhydantoin 81 61 119 16α-Hydroxyprogesterone 83 76 105 17α-Hydroxyprogesterone 83 105 88 7-(β-Hydroxypropyl) theophylline 111 97 110 11β-Hydroxytestosterone 83 230 51 Ibuprofen 69 102 95 Imidapril hydrochloride 97 115 100 Imidazole 109 109 94 Imipramine hydrochloride 23 29 71 Indapamide 69 51 89 Indiplon 71 102 82 3-Indoleacetic acid 91 87 83 Indole-3-carbinol 59 14 73 3-Indoleethanol 83 35 79 Indomethacin 60 69 93 Iodoacetamide 84 113 100 Iproniazid phosphate 115 104 104 Irbesartan 72 81 33 Irinotecan 64 5 100 Isocarboxazid 75 71 94

DL-Isocitric acid 110 106 98 Isoniazid 118 119 100

Isopropyl-β-D(-)-thiogalactopyranoside 108 118 91 JW480 8 9 81 Kaempferol 32 37 6 Ketoconazole 32 26 56 Ketoprofen 88 123 94 Ketotifen fumarate 70 72 63 Labetalol hydrochloride 82 26 90 Lauric acid sodium salt 41 108 96 Leflunomide 63 34 118 Z-Leu-Leu-Leu-al 12 13 63 Leupeptin 90 73 105 Levofloxacin 106 81 109 Lithocholic acid 0 4 2 Loperamide hydrochloride 66 6 50 Lorazepam 18 35 70 Losartan carboxaldehyde 58 21 73 Losartan carboxylic acid 76 34 90 Losartan potassium 83 63 120 Lovastatin 6 6 17 Loxapine succinate salt 20 14 66

D-Mannose-6-phosphate disodium salt 82 97 85 Maprotiline hydrochloride 54 74 71 Medroxyprogesterone acetate 56 50 61 Melatonin 84 63 81 MelQ 78 109 97

L(-)-Menthol 11 31 113 Mephenytoin 84 93 78 Metformin 120 107 107 Methapyrilene hydrochloride 91 100 105 Methimazole 99 86 93 Methotrexate 74 105 92 5-Methoxy-2-methylindole-3-acetic acid 86 83 96 (S)-(+)-6-Methoxy-α-methyl-2-naphthaleneacetic acid 71 87 105 5-Methoxytryptamine hydrochloride 92 108 105 3-Methylflavone-8-carboxylic acid 30 29 63 3-Methylindole 60 41 88

α-Methyl-L-dopa 116 93 108 5-(4-Methylphenyl)-5-phenyl-hydantoin 63 89 108 1-Methyl-4-phenyl-1,2,3,6-tetrahydro-pyridine hydrochloride 93 92 99 4-Methylumbelliferone 79 65 87 Methylviologen 104 102 105 3-Methylxanthine 103 90 112 Metoprolol tartrate 93 53 98 Metyrapon 92 93 80 Mevastatin 8 20 34 Mifepristone 54 45 27 Milnacipran hydrochloride 98 51 113 Monobromobimane 74 79 73 Monomethylol dimethylhydantoin 96 90 102 Montelukast 43 5 16 Mycophenolic acid 92 96 82 Nalidixic acid 99 90 101 Naloxone hydrochloride 57 123 74 α-Naphthoflavone 10 19 14 β-Naphthoflavone 67 79 86 1-Naphthol 12 28 82 2-Naphthol 35 47 74 1,2-Naphthoquinone 0 3 36 1,4-Naphthoquinone 13 17 52 1-Naphthyl acetate 10 7 35 1-Naphthyl isothiocyanate 26 36 102 1-(1-Naphthyl)-2-thiourea 80 52 89 Nateglinide 148 70 106 Nefazodone hydrochloride 63 18 62 Nialamide 92 67 83 Nickel (II) sulfate hexahydrate 97 69 81 (-)-Nicotine 109 93 101 Nicotinic acid N-oxide 102 101 80 Niflumic acid 42 42 95 Nilutamide 65 65 105 Nimesulide 36 93 69 Nimodipine 18 19 62 2-Nitrobenzoic acid 93 103 92 Nordihydroguaiaretic acid 0 0 0

L-(-)-Norepinephrine 94 74 89 Norfloxacin 100 92 71 Nortriptyline hydrochloride 30 52 83 1-Octanesulfonic acid sodium salt 96 77 93 Olmesartan 101 103 102 Omeprazole 90 79 86 Ondansetron hydrochloride 90 85 92 Oxazepam 44 24 101 Oxcarbazepine 57 73 97 Oxibendazole 97 98 91 Oxonic acid potassium salt 89 84 100 Oxybutynin chloride 14 22 68 Oxydized nifedipine 34 22 98 Oxymetholine 23 81 56 Paclitaxel 57 56 75 Paeoniflorin 95 94 91 Palmitic acid 25 41 97 Papaverine 64 51 41 1-Pentanesulfonic acid sodium salt 97 99 94 Pepstatin A 119 116 116 Perphenazine 15 13 36 Phenacetin 67 86 109 1,7-Phenanthroline 78 49 80 Phenazine ethosulfate 101 102 96 p-Phenetidine 69 114 101

L-(-)-Phenylalanine 93 88 110 Phenylbutazone 94 30 87 trans-2-Phenylcyclopropylamine hydrochloride 123 98 100 (R)-1-Phenyl-1,2-ethanediol 85 83 89 (S)-1-Phenyl-1,2-ethanediol 90 70 90 Phenylmethylsulfonyl fluoride 6 9 87 Phenytoin 106 98 87 PhIP hydrochloride 80 64 89

L-α-Phosphatidyl choline dilauroyl 98 146 127 Phthalaldehyde 28 36 70 Pilocarpine hydrochloride 98 89 85 Pioglitazone 36 47 75 Pitavastatin lactone 16 18 63 Pizotifen malate 48 43 70 Polymyxin B sulfate salt 97 86 75 Potassium 4-nitrophenyl sulfate 91 104 92 Potassium ferricyanide 97 83 83 Prasugrel hydrochloride 23 14 13 Pravastatin sodium salt 106 83 94 Praziquantel 71 61 131 Prazosin hydrochloride 77 51 78 Prednisolone 97 87 84 5α-Pregnan-3,20-dione 72 110 76 5β-Pregnan-3,20-dione 17 130 58 5-Pregnen-3β-ol-20-one-16α-carbonitrile 47 108 85 Pregnenolone-16α-carbonitrate 75 77 96 Prilocaine hydrochloride 89 97 82 Probenecid 91 105 97 Probucol 89 90 88 Procainamide hydrochloride 101 98 88 Procaine hydrochloride 81 82 98 Prochlorperazine dimaleate salt 14 19 14 Progesterone 11 108 74 Proglumide 102 105 95 Promazine hydrochloride 57 35 76 Propanil 57 82 107 (R)-(+) Propranolol hydrochloride 93 89 88

DL-Propranolol hydrochloride 93 84 78 6-Propyl-2-thiouracil 119 105 84 Protriptyline hydrochloride 43 65 74 Pyrazinamide 108 97 92 Pyrazinecarboxylic acid 105 95 85 Pyrazole 96 89 85 Pyrene 51 61 82 Pyridoxal 5'-phosphate 95 95 79 Pyrimethamine 98 71 92 Quercetin 19 8 33 Quetiapine hemifumarate 30 19 83 Quinine 109 76 90 Quinine sulfate dihydrate 94 78 86 Qunidine sulfate dihydrate 114 72 87 8-Quinolinol 75 81 86 Raloxifene hydrochloride 0 0 0 Ranitidine hydrochloride 79 91 84 Rebamipide 74 79 89 Resveratrol 40 43 26 Rosiglitazone 13 15 58 Rosuvastatin 78 65 93 Roxithromycin 65 91 72 Rubidium chloride 98 102 103

D-Saccharic acid 1,4-lactone 97 122 99 Semicarbazide hydrochloride 101 94 103 Sennidine A 57 73 94 Sennoside A 93 79 67 Sertraline hydrochloride 7 38 2 Simvastatin 0 0 0 SKF-525A 29 55 61 SN-38 98 84 99 Sodium hydrogen sulfite 92 93 98 Sodium laurylsulfate 21 50 112 Sodium N-lauroyl salcocinate 53 87 111 Sodium orthovanadate 95 96 105 Sodium taurocholate 81 93 107 Sodium tungstate (VI) dihydrate 102 99 103 Sparteine 89 91 81 Spermine 105 100 92 Spironolactone 59 77 69 Strontium chloride hexahydrate 75 95 104 Succinic acid 100 95 102 Sulfamethazine 121 75 75 Sulfamethoxazole 98 92 94 Sulfaphenazole 95 93 96 Sulfasalazine 115 53 96 Sulfobromophthalein sodium salt 53 32 93 5-Sulfosalicylic acid dihydrate 85 93 101 Sulindac 39 49 93 (±)-Sulpiride 107 99 84 Tacrine hydrochloride 83 84 100 Tamoxifen citrate 22 15 101 Tegafur 83 95 64 Telmisartan (50 µM) 86 10 110 Temocapril hydrochloride 81 72 115 Teprenone 35 16 72 Terazosin hydrochloride 95 65 97 Terbinafine 36 2 44 Terbutaline 104 93 97 Terfenadine 22 13 61 Testosterone 36 123 92 Tetrabutylammonium hydrogen sulfate 99 81 88 Tetrabutylammonium hydroxide 88 92 101 Tetracycline 106 95 106 1,2,3,6-Tetrahydro-4-phenylpyridine hydrochloride 96 98 103 Tetrahydroxy tryptamine hydrochloraide 106 113 97 Tetramethyl ammonium chloride 94 94 87 Theobromine 96 96 93 Theophylline 96 93 102 Thiamine hydrochloride 105 96 102 Thioacetamide 95 85 97 Thiobenzamide 105 93 103 Thiourea 78 84 94 Thiouric acid sodium salt dihydrate 89 93 106 Thymidine 91 104 90 Thymine 90 91 101

L-Tryptophan 90 86 94

L-Thyroxine 3 9 3 Ticlopidine hydrochloride 14 19 106 (±)-α-Tocopherol nicotinate 72 79 111 Tolbutamide 70 58 90 Tolcapone 38 38 96 Tolmetin sodium 86 92 100 Tolperisone hydrochloride 40 61 101 Toremifene base 31 7 85 Tranilast 89 17 70 Trazodone hydrochloride 58 28 89 Triacetyloleandomycin 93 52 76 Trichostatin A 23 44 68 Tricine 93 91 102

3,3',5-Triiodo-L-thyronine 4 8 5 Trimipramine maleate 30 63 77 Troglitazone 4 18 17 Trp-P-1 acetate 76 83 51 Tryptamine hydrochloride 96 85 100

L-Tyrosine 83 92 91 U0126 86 26 85 Uracil 93 90 91 Uridine 88 78 107 Uridine 5'-diphosphate sodium salt 99 96 104 Valproic acid sodium salt 93 83 98 Valsartan 86 101 106 Vanillylamine hydrochloride 87 95 99 Verapamil hydrochloride 13 11 36 Vinblastine sulfate 91 9 15 R,S-Warfarin 89 49 97 Wogonin 42 21 14 WST-1 110 99 93 WY-14643 91 27 63 Zafirlukast 15 7 45 Zolpidem 41 65 99 Zomepirac sodium salt 64 92 96

Supplemental Fig. 1. Inhibitory effects of digitonin on fenofibrate hydrolase activities in

HLM and recombinant enzymes. IC50 value was calculated by evaluating the hydrolase activity of fenofibrate (10 µM), which is used for a probe substrate of CES1. Fenofibrate hydrolase activities were determined according to our previous studies (Fukami et al., Drug Metab Dispos 38:2173-2178, 2010). The control fenofibrate hydrolase activities of recombinant CES1 and HLM were 55.4 and 81.9 nmol/min/mg protein, respectively. Each point represents the mean ± SD of triplicate determinations.