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4 protocols using sulfaphenazole

1

In Vitro Enzymatic Characterization

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Sepin-1 (2,2-dimethyl-5-nitro-2H-benzimidazole-1,3-dioxide) with >97% purity was purchased from ChemBridge (San Diego, CA, United States). Phenacetin, efavirenz, paclitaxel, (S)-mephenytoin, quercetin, α-naphthoflavone, ticlopidine, ketoconazole, sulfaphenazole, and dextromethorphan were purchased from Cayman Chemical (Ann Arbor, MI, United States). Reduced GSH, diclofenac sodium, quinidine, midazolam solution, formic acid, and NADPH were obtained from Sigma-Aldrich (St. Louis, MO, United States). HLM, MLM, and RLM, and the recombinant human CYP450s (EasyCYP Bactosomes) were purchased from XenoTech (Lenexa, KS, United States). All the solvents for LC and MS were of the highest grade commercially available.
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2

Cytochrome P450 Enzyme Inhibition Assay

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DAMGO, quinidine, ketoconazole, morphine sulfate, and alprazolam were purchased from MilliporeSigma (St Louis, MO). Mitragynine, 7-hydroxyMitragynine, midazolam, 1′-hydroxymidazolam, sulfaphenazole, and NADPH were obtained from Cayman Chemical Company (Ann Arbor, MI). Diclofenac, 4′-hydroxyDiclofenac, dextromethorphan, and dextrorphan were obtained from Toronto Research Chemicals, Inc. (Toronto, Canada). Potassium phosphate buffer salts were obtained from Fisher Scientific (Fair Lawn, NJ). Human liver microsomes were obtained from XenoTech, LLC (H0604, mixed gender, pool from 15 donors, lot no. 1010191; Kansas City, KS). All other chemicals and reagents used were analytical grade.
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3

In Vitro P450 Inhibition Assay

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2-Hydroxypropyl-β-cyclodextrin and ketoconazole were purchased from Sigma Aldrich (St. Louis, MO). 4-Sulfobutylether-β-cyclodextrin was a gift from Ligand Pharmaceuticals (La Jolla, CA). Pluronics L81 and F127 were purchased from BASF (Ludwigshafen, Germany). d7-Cholesterol was purchased from Avanti Polar Lipids (Alabaster, AL). Baculosome Plus P450 reagent kits and quinidine were purchased from ThermoFischer Scientific (Grace Island, NY). Isoforms chosen were 3A4, 2C9 and 2D6. All reagents were used as received without further purification or characterization. Sulfaphenazole was obtained from Cayman Chemical (Ann Arbor, MI).
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4

Lipid Metabolic Pathway Reagent Procurement

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Tung oil and standard CLA oil were obtained from the Nisshin OilliO Group, Ltd. NADPH, NADP + , NADH, and NAD + were purchased from Oriental Yeast Co., Ltd. (Tokyo, Japan) . Trimethylsilyldiazomethane (TMSN 2 CH 3 ) , trimethylolaminomethane acetate (Tris-actate) , fluvoxamine, ticlopidine, montelukast, fluconazole, chlormethiazole, and ketoconazole were purchased from Tokyo Chemical Industry (Tokyo, Japan) . Indomethacin, niflumic acid, 2-chloroethyl ethyl sulfide (CEES) , 17-octadecynoic acid (17-ODYA) , and HET0016 were purchased from Sigma-Aldrich (St. Louis, MO) . Lauric acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, arachidonic acid, prostaglandin A1, sulfaphenazole, and quinidine were obtained from Cayman Chemical Company (Ann Arbor, MI) . All other chemicals (sucrose, EDTA・2Na・2H 2 O, DTT, PMSF) were purchased from Wako Pure Chemical Industries, Ltd.
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