Ugt1a8

Ib, Ib glucuronide and irbesartan (internal standard, IS) were supplied by the Department of Medicinal Chemistry of China Pharmaceutical University, Nanjing, China. UPDGA (63700-19-6), d-saccharic acid 1,4-lactone (61278-30-6), alamethicin (27061-78-5), BSA (9048-46-8), NADP (53-59-8), G-6-P (3671-99-6), and PDH (9001-40-5) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Tris-Base were purchased from Biosharp (Seoul, South Korea).
Mixed Gender Pooled Human Liver Microsomes (HLM, from 25 subjects of 25–89 years age), Mixed Gender Pooled Cynomolgus monkey Liver Microsomes (MKLM, from 14 subjects of 4–5 years age), Mixed Gender Pooled Beagle Dog Liver Microsomes (DLM, from 7 subjects of 11 weeks age), Mixed Gender Pooled Sprague-Dawley Rat Liver Microsomes (RLM, from 100 subjects of 2–3 months age) and Mixed Gender Pooled CD1 Mice Liver Microsomes (MLM, from 500 subjects of 5–8 weeks age) were purchased from the Research Institute for Liver Diseases (Shanghai) Co. Ltd. (Shanghai, China). Supersome preparations containing recombinant UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, UGT1A10, UGT2B4, UGT2B7, UGT2B15 or UGT2B17 (expressed in baculovirus-infected insect cells) and UGT Inset Cell Control Supersomes^TM were purchased from BD Biosciences (San Jose, CA, USA). All other chemicals and solvents used were of the highest quality and analytical grade or higher.

Bavachalcone and corylin were purchased from Shifeng Corp. (Shanghai, China), and their purities were all above 98%. 4-Methylumbelliferone(4-MU), 4-methylumbelliferone-β-D-glucuronide(4-MUG), Tris-HCl, alamethicin, 7-hydroxycoumarin, and uridine 5′-diphosphoglucuronic acid (UDPGA) (trisodium salt) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Recombinant human UGT supersomes (UGT1A1, UGT1A3, UGT1A7, UGT1A8, UGT1A10, and UGT2B4) expressed in baculovirus-infected insect cells were obtained from BD Gentest Corp. (Woburn, MA, USA). Solvents and other reagents were of HPLC grade.

IMM-H004 (purity >99%), M1, and M2 (purity >90%) (Li et al., 2010 (link)) were synthesized by Laboratory of Chemical Synthesis, and IMM-H004G (M3) (purity >99%) for animal study was provided by Prof. Dai (Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, China). Propranolol (internal standard, IS), β-glucuronidase, sulfatase, 3′-phosphoadenosine-5′-phosphosulfate (PAPS), uridine 5′-diphosphoglucuronic acid (UDPGA), and alamethicin were obtained from Sigma-Aldrich (St. Louis, MO). Rat liver microsomes (RLMs) and cytosols were prepared by differential ultracentrifugation, and the protein concentration was determined by bicinchoninic acid assay (Beyotime Institute of Biotechnology, Jiangsu, China). Pooled mixed-gender human liver microsomes (HLMs), human liver cytosols, recombinant human cytochrome P450 enzymes (CYP1A1, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP2J2, CYP3A4, CYP4A11, CYP4F2, and CYP4F3), recombinant human UDP-glucuronosyltransferases (UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, UGT1A10, UGT12B4, UGT12B7, UGT12B10, UGT12B15, and UGT12B17), and recombinant human sulfotransferase (SULT1A1, SULT1A2, SULT1A3, SULT1B1, SULT1C2, SULT1C4, SULT1E1, and SULT2A1) were purchased from BD Gentest (Woburn, MA).