Cyp2a6

The methanolic extract of E. longifolia roots was provided by the Institute of Marine Biochemistry, Vietnam Academy of Science and Technology (Hanoi, Vietnam). Pooled human liver microsomes and recombinant CYP1A2, CYP2A6, and CYP2C19 isozymes were purchased from BD Gentest (Woburn, MA, USA). Glucose-6-phosphate, β-NADP⁺, Glucose-6-phosphate dehydrogenase, phenacetin, coumarin, diclofenac, mephenytoin, dextromethorphan, midazolam, ketoconazole, and terfenadine were obtained from Sigma Chemical Co. (Saint Louis, MO, USA). All other solvents used were of HPLC grade and were purchased from J. T. Baker (Phillipsburg, NJ, USA). Distilled water was prepared using a Milli-Q purification system (Millipore, Billerica, MA, USA).

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).