2.1 Chemicals and reagents.
Ningetinib, M1, deuterated ningetinib (D6-ningetinib) and deuterated M1 (D6-M1) were kindly provided by Sunshine Lake Pharma Co., Ltd. (Guangdong, China). Digoxin, imatinib, gefitinib and vincristine were purchased from Dalian Meilun Biotechnology Co., Ltd. (Liaoning, China). Human recombinant CYP450 enzymes (CYP1A1, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP2J2, CYP3A4, CYP3A5, CYP4A11 and CYP4F2) and pooled human intestinal microsomes (HIMs), human kidney microsomes (HKMs) and human lung microsomes (HLUMs) were obtained from BD Gentest (Woburn, MA). Pooled human liver microsomes (HLMs) were supplied by Reid Liver Disease Research Co., Ltd. (Shanghai, China). Primary human hepatocytes were obtained from Sekisui XenoTech (Kansas City, Kansas). Nicotinamide adenine dinucleotide phosphate (NADPH), GF120918, KO143 and sodium carboxymethyl cellulose (CMC-Na) were purchased from Sigma-Aldrich (St. Louis, MO). TAK-875 was obtained from Shanghai Biochempartner Co., Ltd. (Shanghai, China). William’s Medium E (WME), DMEM medium, foetal bovine serum (FBS), L-glutamine, non-essential amino acids, 0.25% trypsin, penicillin G and streptomycin were purchased from Invitrogen (Carlsbad, CA). Deionised water was generated using a Millipore Milli-Q gradient water purification system (Molsheim, France). The Madin-Darby canine kidney (MDCKII)-MDR1 (P-gp transfected cells), MDCKII-BCRP and MDCKII-MRP2 cells were kindly provided by Professor Hongjian Zhang from the School of Pharmacy, Soochow University (Jiangsu, China).