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4 protocols using tienilic acid

1

Chemical Compound Acquisition for Research

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Erythromycin, clarithromycin, roxithromycin, diltiazem, ritonavir, ketoconazole, fluconazole, cimetidine, ticlopidine, omeprazole, fluvoxamine, paroxetine, fluoxetine, quinidine, aflatoxin B1, and sterigmatocystin were obtained from Wako, Japan. Gestodene and mifepristone were obtained from Tokyo Chemical Industry, Japan. G418 was obtained from Funakoshi, Japan. Ethinylestradiol, rifampicin, tienilic acid, terbinafine, hypoxanthine, aminopterin, and thymidine (HAT) and 4,6-diamidino-2-phenylindole (DAPI) were obtained from Sigma Aldrich, USA. All other chemicals were of the highest grade commercially available.
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2

Sauchinone Purity Characterization and Evaluation

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Sauchinone (over 98% purity) was provided by Professor Chin Y.-W. (Pharmacognosy Laboratory of Dongguk University, Seoul, Korea) according to the modification of previously reported method [48 (link)]. Hydroxybupropion, 4-hydroxymephenytoin, 6-hydroxychlorzoxazone, S-mephenytoin, and pooled human liver microsomes from a mixed pool of 50 donors (HLMs; BD Ultra Pool HLM 50, cat. 452156) were purchased from BD Gentest Co. (Woburn, MA, USA). Acetaminophen, chlorzoxazone, clopidogrel, coumarin, furafylline, 6α-hydroxytestosterone, 7-hydroxycoumarin, ketoconazole, 4-methylpyrazole, omeprazole, paroxetine, phenacetin, quinidine, sibutramine, sulfaphenazole, testosterone, tetraethylthiuram disulfide, ticlopidine, tienilic acid, tolbutamide, tranylcypromine, verapamil, and reduced form of NADPH (as a tetrasodium salt) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dextrorphan and 4-hydroxytolbutamide were purchased from Toronto Research Chemicals Inc. (North York, ON, Canada). Bupropion, carbamazepine (internal standard, IS), dextromethorphan, fluvoxamine, fluoxetine, and 8-methoxypsoralen were purchased Wako Co. (Tokyo, Japan). All other chemicals and reagents used were of analytical grade.
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3

Synthesis and Characterization of Brepocitinib Metabolites

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General Chemicals. Brepocitinib and metabolites M1, M2, M3 and M4 were synthesized at Pfizer Inc. (Groton, CT). All chemicals were of analytical grade or better and included the following: Girard's Regent P from TCI chemicals (Portland, OR), L-Ascorbic acid from Sigma Aldrich (St. Louis, MO). Citric Acid, Monohydrate from J.T. Baker (Avantor, Radnor, PA). MgCl 2 , NADPH, diclofenac, furafylline, tienilic acid, esomeprazole, quinidine, 1aminobenzotriazole, 1 M potassium phosphate dibasic solution, 1 M potassium phosphate monobasic solution, sodium bicarbonate, and dimethyl sulfoxide (DMSO) were purchased from Sigma Aldrich (St. Louis, MO). Troleandomycin was purchased from Fischer Scientific (Waltham, MA). HPLC solvents included ammonium acetate, ACN, MeOH, and formic acid from Biosolve (Valkenswaard, Netherlands); trifluoracetic acid from Sigma Aldrich (Amsterdam, Netherlands); isopropyl amine from iPAm, Merck (Amsterdam, Netherlands); 2 mM ammonium acetate/MeOH/ACN (95/2.5/2.5% v/v) from Brand-Nu Laboratories (Meriden, CT) and MeOH, ACN, and water from J.T. Baker (Avantor, Radnor, PA). Pooled HLM 20 .0 mg of protein per ml, 0.31 nmol of cytochrome P450 (CYP) per mg of protein] were prepared and characterized by Sekisui Xenotech (Kansas City, KS) from 50 individual human donors of both sexes. Human whole blood and plasma were purchased from BioIVT (Baltimore,
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4

Preparation of Stock Solutions for Cell Culture

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Acetaminophen, tert-butylhydroquinone, and tienilic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Aminopyrine, sulfamethoxazole, and forskolin were purchased from Fujifilm Wako Pure Chemical Co. (Osaka, Japan). Erythromycin, valsartan, and warfarin were purchased from LKT Laboratories, Inc. (St Paul, MN, USA). Nevirapine, pravastatin, and procainamide were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Stock solutions of all test drugs were prepared in dimethyl sulfoxide (DMSO) at a concentration of 200 mM. Each solution was adequately diluted with Dulbecco's modified Eagle's medium (DMEM) (Nissui Pharmaceutical Co., Ltd., Tokyo, Japan) so that the final concentration of DMSO in the culture medium was 0.5%.
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