Uridine diphosphate glucuronic acid

Briefly, 1,25(OH)₂D₃, ACET, acetaminophen glucuronide (AG), BUP, hydroxybupropion (HBUP), carbamazepine (CARB), diltiazem (DIL), theophylline (THEO), TOL, 4-hydroxytolbutamide (HTOL), and uridine diphosphate glucuronic acid (UDPGA) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Dihydronicotinamide adenine dinucleotide phosphate (NADPH) was purchased from Corning, Inc. (New York, NY, USA). All solvents used were of HPLC grade. The PrimeScriptTM 1st strand cDNA Synthesis Kit and SYBR^® Premix Ex TaqTM II ROX plus were purchased from Takara Bio, Inc. (Shiga, Japan). Forward and reverse primers for quantitative polymerase chain reaction (qPCR) experiment were synthesized by Bioneer Co. (Daejeon, Korea).

α-MG was purified (> 96.0% purity) in the College of Pharmacy, Dongguk University (Seoul, South Korea) according to the previously reported protocol [34 (link)]. Docetaxel [internal standard (IS) for HPLC-MS/MS; > 99% purity] was supplied from Shin Poong Pharmaceutical Company, Ltd (Ansan, South Korea). Polyethylene glycol 400 (PEG 400) was obtained from the Showa Chemical Company (Tokyo, Japan). Dextran, the reduced form of β-nicotinamide adenine dinucleotide phosphate (NADPH; as a tetrasodium salt), uridine diphosphate glucuronic acid (UDPGA; as a trisodium salt) and Tris-buffer were purchased from Sigma-Aldrich (St. Louis, MO, USA). Methanol, acetonitrile, formic acid and water were purchased from Fisher Scientific (Seoul, South Korea). All other chemicals and reagents used were of analytical grade.

Saxagliptin, trelagliptin, and gemigliptin (Figure 1) were purchased from AdooQ Bioscience (>98%, Irvine, CA, USA); linagliptin and 5-hydroxySaxagliptin from Toronto Research Chemicals (Toronto, Canada); ammonium acetate (99.99%), β-nicotinamide adenine dinucleotide phosphate sodium salt hydrate (NADP⁺), dimethyl sulfoxide (DMSO, >99.7%), glucose-6-phosphate (G6P), glucose-6-phosphate dehydrogenase (G6PD), L-cysteine, magnesium chloride, reduced glutathione (GSH, ≥99%), 3′-phosphoadenosine-5′-phosphosulfate (PAPS), potassium phosphate, uridine diphosphate glucuronic acid (UDPGA, >98%), and vildagliptin from Sigma-Aldrich (St. Louis, MO, USA); and rat liver microsomes (RLM, R1000) from XenoTech (Lenexa, KS, USA). All solvents were LC-MS grade from Fisher Scientific (Pittsburgh, PA, USA).

Human UGT1A9-overexpressing HeLa cells (HeLa1A9 cells) were established as described previously (Jiang et al., 2011 (link)), and were provided by Prof. Baojian Wu who works in Jinan University in Guangzhou of China. Alamethicin, D-saccharic-1,4-lactone, magnesium chloride (MgCl₂), MK571, Ko143, propofol, and uridine diphosphate glucuronic acid (UDPGA) were purchased from Sigma-Aldrich (St Louis, MO, United States), whereas propofol-O-glucuronide from Toronto Research Chemicals (North York, ON, Canada). Fraxetin was provided by Guangzhou Fans Biotechnology Co., Ltd., (Guangzhou, China), and human UGT1A9 was purchased from Corning Biosciences (New York, NY, United States). UGT1A9 antibody was purchased from BD Biosciences (Woburn, MA, United States). The anti-BCRP, anti-MRP1, anti-MRP2, anti-MRP3, and anti-MRP4 antibodies were purchased from OriGene Technologies (Rockville, MD, United States). All other chemicals and reagents (analytical grade or better) were commercially available.

Uridine diphosphate glucuronic acid (UDPGA), magnesium chloride (MgCl₂), alamethicin and d-saccharic-1,4-lactone were provided by Sigma-Aldrich (St. Louis, MO, USA). Pooled human liver microsomes (HLM), human intestinal microsomes (HIM), recombinant expressed human UGT Supersomes™ (UGT1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 2B4, 2B7, 2B10, 2B15 and 2B17), dog liver microsomes (DLM), rat liver microsomes (RLM), monkey liver microsomes (MkLM), rabbit liver microsomes (RaLM) and guinea pig liver microsomes (GpLM) were all obtained from Corning Biosciences (Corning, Corning, NY, USA). Wushanicaritin (purity > 98%) was purchased from Jingzhu Medical Technology Co., Ltd. (Nanjing, China). Wushanicaritin 3-O-glucuronidation (G1) and 7-O-glucuronidation (G2) were prepared according to the previous study [12 (link)]. The detailed NMR data of G1 and G2 are shown in Table S1, and their NMR spectra were displayed in Figures S1 and S2. β-estradiol, chenodeoxycholic acid (CDCA), propofol and zidovudine (AZT) were purchased from Aladdin Chemicals (Shanghai, China). All other chemicals and reagents were of analytical grade or the highest grade commercially available.