Evaluation of 309 molecules as inducers of CYP3A4, CYP2B6, CYP1A2, OATP1B1, OCT1, MDR1, MRP2, MRP3 and BCRP in cryopreserved human hepatocytes in sandwich culture

• Published in: Xenobiotica Vol. 45; no. 2; pp. 177 - 187
• Main Authors: Badolo, Lassina, Jensen, Bente, Säll, Carolina, Norinder, Ulf, Kallunki, Pekka, Montanari, Dino
• Format: Journal Article
• Language: English
• Published: England Informa UK Ltd 01-02-2015; Informa Healthcare
• Subjects: ATP Binding Cassette Transporter, Sub-Family G, Member 2; ATP-Binding Cassette Transporters - chemistry; ATP-Binding Cassette Transporters - metabolism; ATP-Binding Cassette, Sub-Family B, Member 1 - chemistry; ATP-Binding Cassette, Sub-Family B, Member 1 - metabolism; Cell Culture Techniques; cryopreserved human hepatocytes; Cytochrome P-450 CYP1A2 Inducers - chemistry; Cytochrome P-450 CYP1A2 Inducers - pharmacology; Cytochrome P-450 CYP2B6 Inducers - chemistry; Cytochrome P-450 CYP2B6 Inducers - pharmacology; Cytochrome P-450 CYP3A - metabolism; Cytochrome P-450 CYP3A Inducers - chemistry; Cytochrome P-450 CYP3A Inducers - pharmacology; Cytochrome P450; Drug Discovery - methods; drug-drug interactions; hepatic transporters; Hepatocytes - drug effects; Hepatocytes - enzymology; Humans; induction; Multidrug Resistance-Associated Proteins - chemistry; Multidrug Resistance-Associated Proteins - metabolism; Neoplasm Proteins - chemistry; Neoplasm Proteins - metabolism; Organic Anion Transporters - chemistry; Organic Anion Transporters - metabolism; Organic Cation Transporter 1 - chemistry; Organic Cation Transporter 1 - metabolism; sandwich culture; Solute Carrier Organic Anion Transporter Family Member 1b1; Structure-Activity Relationship; cryopreserved human hepatocytes; sandwich culture; hepatic transporters; induction; Cytochrome P450; drug–drug interactions
• ISSN: 0049-8254; 1366-5928
• DOI: 10.3109/00498254.2014.955831

Abstract
1. Regulation of hepatic metabolism or transport may lead to increase in drug clearance and compromise efficacy or safety. In this study, cryopreserved human hepatocytes were used to assess the effect of 309 compounds on the activity and mRNA expression (using qPCR techniques) of CYP1A2, CYP2B6 and CYP3A4, as well as mRNA expression of six hepatic transport proteins: OATP1B1 (SCLO1B1), OCT1 (SLC22A1), MDR1 (ABCB1), MRP2 (ABCC2), MRP3 (ABCC3) and BCRP (ABCG2).
2. The results showed that 6% of compounds induced CYP1A2 activity (1.5-fold increase); 30% induced CYP2B6 while 23% induced CYP3A4. qPCR data identified 16, 33 or 32% inducers of CYP1A2, CYP2B6 or CYP3A4, respectively. MRP2 was induced by 27 compounds followed by MDR1 (16) > BCRP (9) > OCT1 (8) > OATP1B1 (5) > MRP3 (2).
3. CYP3A4 appeared to be down-regulated (≥2-fold decrease in mRNA expression) by 53 compounds, 10 for CYP2B6, 6 for OCT1, 4 for BCRP, 2 for CYP1A2 and OATP1B1 and 1 for MDR1 and MRP2.
4. Structure-activity relationship analysis showed that CYP2B6 and CYP3A4 inducers are bulky lipophilic molecules with a higher number of heavy atoms and a lower number of hydrogen bond donors. Finally, a strategy for testing CYP inducers in drug discovery is proposed.