低清除率药物的代谢稳定性预测模型研究进展
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摘要
药物代谢的稳定性测试是新药发现阶段的关键环节,实现药物的低清除率通常是药物代谢稳定性设计中的重要目标。如何准确评估低清除率药物的代谢稳定性参数,并用体外代谢数据预测人体药动学已经成为新药研发阶段的挑战。传统的肝微粒体模型和悬浮肝细胞模型的孵育时间短,低清除率药物无法产生足够的代谢转化,因此进一步模拟体内环境和延长肝细胞培养时间的新型模型逐渐发展起来。本文重点介绍了新型的低清除率药物代谢稳定性预测模型的原理和优缺点等,包括肝细胞传递式培养模型、单层贴壁肝细胞培养模型、共培养模型和微灌流模型等,同时对模型的发展趋势进行展望,以期为早期先导化合物的代谢稳定性检测提供借鉴和优化。
The metabolic stability test of drugs is a key step in drug discovery and achieving low clearance is frequently the goal in the design of drug.Increased drug metabolism stability can reduce drug dosage,enhance drug exposure and prolong drug half-life.Accurately assessing the metabolic stability parameters of low clearance drugs and predicting human pharmacokinetics has become a challenge.Traditional tools in vitro including microsomes and suspended primary hepatocytes are limited by incubation time,which is not long enough to make sufficient metabolic conversion.Determination of intrinsic clearance or metabolic pathways and mechanisms of drug are implicated.Novel models tend to further mimic the in vivo environment in order to prolong lifetime of hepatocytes and achieve sufficient metabolic turnover of drugs for monitoring.In vitro-in vivo correlation of intrinsic clearance of methodologies has evaluated to support the reliability in predicting human pharmacokinetics.Application of these methodologies greatly decreases the forthputting of experimental animals and the release of expensive clinical trials during the acquisition of pharmacokinetic parameters.In this review,we summarized the principles,advantages and disadvantages of the novel in vitro methodologies for metabolic stability dealing with low-turnover drugs,including hepatocyte relay method,plated human hepatocytes,coculture system and microfluidic devices.Future prospect is proposed for in vitro metabolic models and it provides reference and optimization in metabolic stability for early lead compounds.
The metabolic stability test of drugs is a key step in drug discovery and achieving low clearance is frequently the goal in the design of drug.Increased drug metabolism stability can reduce drug dosage,enhance drug exposure and prolong drug half-life.Accurately assessing the metabolic stability parameters of low clearance drugs and predicting human pharmacokinetics has become a challenge.Traditional tools in vitro including microsomes and suspended primary hepatocytes are limited by incubation time,which is not long enough to make sufficient metabolic conversion.Determination of intrinsic clearance or metabolic pathways and mechanisms of drug are implicated.Novel models tend to further mimic the in vivo environment in order to prolong lifetime of hepatocytes and achieve sufficient metabolic turnover of drugs for monitoring.In vitro-in vivo correlation of intrinsic clearance of methodologies has evaluated to support the reliability in predicting human pharmacokinetics.Application of these methodologies greatly decreases the forthputting of experimental animals and the release of expensive clinical trials during the acquisition of pharmacokinetic parameters.In this review,we summarized the principles,advantages and disadvantages of the novel in vitro methodologies for metabolic stability dealing with low-turnover drugs,including hepatocyte relay method,plated human hepatocytes,coculture system and microfluidic devices.Future prospect is proposed for in vitro metabolic models and it provides reference and optimization in metabolic stability for early lead compounds.
引文
[1] Obach RS.Predicting clearance in humans from in vitro data[J].Curr Top Med Chem,2011,11(4):334-339.
[2] Di L,Obach RS.Addressing the challenges of low clearance in drug research[J].AAPS J,2015,17(2):352-357.
[3] Di L,Trapa P,Obach RS,et al.A novel relay method for determining low-clearance values[J].Drug Metab Dispos,2012,40(9):1860-1865.
[4] Grime KH,Barton P,McGinnity DF.Application of in silico,in vitro and preclinical pharmacokinetic data for the effective and efficient prediction of human pharmacokinetics[J].Mol Pharm,2013,10(4):1191-1206.
[5] Sohlenius-Sternbeck AK,Jones C,Ferguson D,et al.Practical use of the regression offset approach for the prediction of in vivo intrinsic clearance from hepatocytes[J].Xenobiotica,2012,42(9):841-853.
[6] Chao P,Uss AS,Cheng KC.Use of intrinsic clearance for prediction of human hepatic clearance[J].Exp Opin Drug Metab Toxicol,2010,6(2):189-198.
[7] Sun LN,Ding L,Yan ZY,et al.Determination the inhibitory potency of bencycloquidium bromide on rat liver cytochrome P450 by LC-MS/MS[J].J China Pharm Univ(中国药科大学学报),2013,44(2):134-140.
[8] Lin C,Ballinger KR,Khetani SR.The application of engineered liver tissues for novel drug discovery[J].Exp Opin Drug Discov,2015,10(5):519-540.
[9] Rodriguez-Antona C,Donato MT,Boobis A,et al.Cytochrome P450 expression in human hepatocytes and hepatoma cell lines:molecular mechanisms that determine lower expression in cultured cells[J].Xenobiotica,2002,32(6):505-520.
[10] Jones HM,Houston JB.Substrate depletion approach for determining in vitro metabolic clearance:time dependencies in hepatocyte and microsomal incubations[J].Drug Metab Dispos,2004,32(9):973-982.
[11] Di L,Atkinson K,Orozco CC,et al.In vitro-in vivo correlation for low-clearance compounds using hepatocyte relay method[J].Drug Metab Dispos,2013,41(12):2018-2023.
[12] Ballard TE,Orozco CC,Obach RS.Generation of major human excretory and circulating drug metabolites using a hepatocyte relay method[J].Drug Metab Dispos,2014,42(5):899-902.
[13] Yang X,Atkinson K,Di L.Novel cytochrome P450 reaction phenotyping for low-clearance compounds using the hepatocyte relay method[J].Drug Metab Dispos,2016,44(3):460-465.
[14] Peng CC,Doshi U,Prakash C,et al.A novel plated hepatocyte relay assay (PHRA) for in vitro evaluation of hepatic metabolic clearance of slowly metabolized compounds[J].Drug Metab Lett,2016,10(1):3-15.
[15] Bonn B,Svanberg P,Janefeldt A,et al.Determination of human hepatocyte intrinsic clearance for slowly metabolized compounds:comparison of a primary hepatocyte/stromal cell co-culture with plated primary hepatocytes and HepaRG[J].Drug Metab Dispos,2016,44(4):527-533.
[16] Griffin SJ,Houston JB.Prediction of in vitro intrinsic clearance from hepatocytes:comparison of suspensions and monolayer cultures[J].Drug Metab Dispos,2005,33(1):115-120.
[17] Blanchard N,Alexandre E,Abadie C,et al.Comparison of clearance predictions using primary cultures and suspensions of human hepatocytes[J].Xenobiotica,2005,35(1):1-15.
[18] Smith CM,Nolan CK,Edwards MA,et al.A comprehensive evaluation of metabolic activity and intrinsic clearance in suspensions and monolayer cultures of cryopreserved primary human hepatocytes[J].J Pharm Sci,2012,101(10):3989-4002.
[19] Hoffmaster KA,Turncliff RZ,LeCluyse EL,et al.P-glycoprotein expression,localization,and function in sandwich-cultured primary rat and human hepatocytes:relevance to the hepatobiliary disposition of a model opioid peptide[J].Pharm Res,2004,21(7):1294-1302.
[20] Kanda K,Takahashi R,Yoshikado T,et al.Total hepatocellular disposition profiling of rosuvastatin and pitavastatin in sandwich-cultured human hepatocytes[J].Drug Metab Pharmacokinet,2018,33(3):164-172.
[21] Swift B,Pfeifer ND,Brouwer KL.Sandwich-cultured hepatocytes:an in vitro model to evaluate hepatobiliary transporter-based drug interactions and hepatotoxicity[J].Drug Metab Rev,2010,42(3):446-471.
[22] Brown JH,Das P,DiVito MD,et al.Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro[J].Acta Biomater,2018,73(4):217-227.
[23] Oorts M,Keemink J,Deferm N,et al.Extra collagen overlay prolongs the differentiated phenotype in sandwich-cultured rat hepatocytes[J].J Pharmacol Toxicol Methods,2018,90(6):31-38.
[24] Keemink J,Oorts M,Annaert P.Primary hepatocytes in Sandwich culture[J].Methods Mol Biol,2015,1250(7):175-188.
[25] Lancett P,Williamson B,Barton P,et al.Development and characterization of a human hepatocyte low intrinsic clearance assay for use in drug discovery[J].Drug Metab Dispos,2018,46(8):1169-1178.
[26] Treijtel N,Barendregt A,Freidig AP,et al.Modeling the in vitro intrinsic clearence of the slowly metabolized compound tolbutamide determined in sandwich-cultures rat hepatocytes[J].Drug Metab Dispos,2004,32:884-891.
[27] Liu Y,Wei J,Lu J,et al.Micropatterned coculture of hepatocytes on electrospun fibers as a potential in vitro model for predictive drug metabolism[J].Mater Sci Eng C Mater Biol Appl,2016,63:475-484.
[28] Oda H,Yoshida Y,Kawamura A,et al.Cell shape,cell-cell contact,cell-extracellular matrix contact and cell polarity are all required for the maximum induction of CYP2B1 and CYP2B2 gene expression by phenobarbital in adult rat cultured hepatocytes[J].Biochem Pharmacol,2008,75(5):1209-1217.
[29] Xiao W,Perry G,Komori K,et al.New physiologically-relevant liver tissue model based on hierarchically cocultured primary rat hepatocytes with liver endothelial cells[J].Integr Biol (Camb),2015,7(11):1412-1422.
[30] Kostadinova R,Boess F,Applegate D,et al.A long-term three dimensional liver co-culture system for improved prediction of clinically relevant drug-induced hepatotoxicity[J].Toxicol Appl Pharmacol,2013,268(1):1-16.
[31] Khetani SR,Bhatia SN.Microscale culture of human liver cells for drug development[J].Nat Biotechnol,2008,26(1):120-126.
[32] March S,Ramanan V,Trehan K,et al.Micropatterned coculture of primary human hepatocytes and supportive cells for the study of hepatotropic pathogens[J].Nat Protoc,2015,10(12):2027-2053.
[33] Chan TS,Yu H,Moore A,et al.Meeting the challenge of predicting hepatic clearance of compounds slowly metabolized by cytochrome P450 using a novel hepatocyte model,HepatoPac[J].Drug Metab Dispos,2013,41(12):2024-2032.
[34] Liu YW,Li HN,Yan SL,et al.Hepatocyte cocultures with endothelial cells and fibroblasts on micropatterned fibrous mats to promote liver-specific functions and capillary formation capabilities[J].Biomacromolecules,2014,15(3):1044-1054.
[35] Ukairo O,Kanchagar C,Moore A,et al.Long-term stability of primary rat hepatocytes in micropatterned cocultures[J].J Biochem Mol Toxicol,2013,27(3):204-212.
[36] Shen C,Xu XM,Meng Q,et al.Studies on rifampicin and isoniazid-induced hepatotoxicity[J].J China Pharm Univ(中国药科大学学报),2005,36(3):250-230.
[37] Wang WW,Khetani SR,Krzyzewski S,et al.Assessment of a micropatterned hepatocyte coculture system to generate major human excretory and circulating drug metabolites[J].Drug Metab Dispos,2010,38(10):1900-1905.
[38] Ramsden D,Tweedie DJ,Chan TS,et al.Bridging in vitro and in vivo metabolism and transport of faldaprevir in human using a novel cocultured human hepatocyte system,HepatoPac[J].Drug Metab Dispos,2014,42(3):394-406.
[39] Ramsden D,Tweedie DJ,St George R,et al.Generating an in vitro-in vivo correlation for metabolism and liver enrichment of a hepatitis C virus drug,faldaprevir,using a rat hepatocyte model (HepatoPac)[J].Drug Metab Dispos,2014,42(3):407-414.
[40] Hutzler JM,Ring BJ,Anderson SR.Low-turnover drug molecules:a current challenge for drug metabolism scientists[J].Drug Metab Dispos,2015,43(12):1917-1928.
[41] Chao P,Maguire T,Novik E,et al.Evaluation of a microfluidic based cell culture platform with primary human hepatocytes for the prediction of hepatic clearance in human[J].Biochem Pharmacol,2009,78(6):625-632.
[42] Novik E,Maguire TJ,Chao P,et al.A microfluidic hepatic coculture platform for cell-based drug metabolism studies[J].Biochem Pharmacol,2010,79(7):1036-1044.
[43] Hultman I,Vedin C,Abrahamsson A,et al.Use of HμREL human coculture system for prediction of intrinsic clearance and metabolite formation for slowly metabolized compounds[J].Mol Pharm,2016,13(8):2796-2807.
[44] Dash A,Inman W,Hoffmaster K,et al.Liver tissue engineering in the evaluation of drug safety[J].Exp Opin Drug Metab Toxicol,2009,5(10):1159-1174.
[45] Vivares A,Salle-Lefort S,Arabeyre-Fabre C,et al.Morphological behaviour and metabolic capacity of cryopreserved human primary hepatocytes cultivated in a perfused multiwell device[J].Xenobiotica,2015,45(1):29-44.
[46] Darnell M,Ulvestad M,Ellis E,et al.In vitro evaluation of major in vivo drug metabolic pathways using primary human hepatocytes and HepaRG cells in suspension and a dynamic three-dimensional bioreactor system[J].J Pharmacol Exp Ther,2012,343(1):134-144.
[47] Darnell M,Schreiter T,Zeilinger K,et al.Cytochrome P450-dependent metabolism in HepaRG cells cultured in a dynamic three-dimensional bioreactor[J].Drug Metab Dispos,2011,39(7):1131-1138.
[48] Antherieu S,Chesne C,Li R,et al.Stable expression,activity,and inducibility of cytochromes P450 in differentiated HepaRG cells[J].Drug Metab Dispos,2010,38(3):516-525.
[49] Davies B,Morris T.Physiological parameters in laboratory animals and humans[J].Pharm Res,1993,10(7):1093-1095.
[50] U.S.Food and Drug Administration.Guidance for industry:Safety Testing of Drug Metabolites[EB/OL].(2016-11-22)[2018-12-17].https://www.fda.gov/Drugs/Guidanc- eComplianceRegulatoryInfo-rmation/Guidances/ucm065014.htm.
[51] European Medicines Agency.Guideline on the investigation of drug interactions[EB/OL].(2012-06-21)[2018-12-17]https://www.ema.europa.eu/en/search/search?search_api_views_fulltext=Guideline%-20on%20the%20investigation%20of% 20drug%20interactions.
[52] Matthew Hutzler J,Linder CD,Melton RJ,et al.In vitro-in vivo correlation and translation to the clinical outcome for CJ-13,610,a novel inhibitor of 5-lipoxygenase[J].Drug Metab Dispos,2010,38(7):1113-1121.
[53] Zientek MA,Youdim K.Reaction phenotyping:advances in the experimental strategies used to characterize the contribution of drug-metabolizing enzymes[J].Drug Metab Dispos,2015,43(1):163-181.
[54] Oda S,Fukami T,Yokoi T,et al.A comprehensive review of UDP-glucuronosyltransferase and esterases for drug development[J].Drug Metab Pharmacokinet,2015,30(1):30-51.
[2] Di L,Obach RS.Addressing the challenges of low clearance in drug research[J].AAPS J,2015,17(2):352-357.
[3] Di L,Trapa P,Obach RS,et al.A novel relay method for determining low-clearance values[J].Drug Metab Dispos,2012,40(9):1860-1865.
[4] Grime KH,Barton P,McGinnity DF.Application of in silico,in vitro and preclinical pharmacokinetic data for the effective and efficient prediction of human pharmacokinetics[J].Mol Pharm,2013,10(4):1191-1206.
[5] Sohlenius-Sternbeck AK,Jones C,Ferguson D,et al.Practical use of the regression offset approach for the prediction of in vivo intrinsic clearance from hepatocytes[J].Xenobiotica,2012,42(9):841-853.
[6] Chao P,Uss AS,Cheng KC.Use of intrinsic clearance for prediction of human hepatic clearance[J].Exp Opin Drug Metab Toxicol,2010,6(2):189-198.
[7] Sun LN,Ding L,Yan ZY,et al.Determination the inhibitory potency of bencycloquidium bromide on rat liver cytochrome P450 by LC-MS/MS[J].J China Pharm Univ(中国药科大学学报),2013,44(2):134-140.
[8] Lin C,Ballinger KR,Khetani SR.The application of engineered liver tissues for novel drug discovery[J].Exp Opin Drug Discov,2015,10(5):519-540.
[9] Rodriguez-Antona C,Donato MT,Boobis A,et al.Cytochrome P450 expression in human hepatocytes and hepatoma cell lines:molecular mechanisms that determine lower expression in cultured cells[J].Xenobiotica,2002,32(6):505-520.
[10] Jones HM,Houston JB.Substrate depletion approach for determining in vitro metabolic clearance:time dependencies in hepatocyte and microsomal incubations[J].Drug Metab Dispos,2004,32(9):973-982.
[11] Di L,Atkinson K,Orozco CC,et al.In vitro-in vivo correlation for low-clearance compounds using hepatocyte relay method[J].Drug Metab Dispos,2013,41(12):2018-2023.
[12] Ballard TE,Orozco CC,Obach RS.Generation of major human excretory and circulating drug metabolites using a hepatocyte relay method[J].Drug Metab Dispos,2014,42(5):899-902.
[13] Yang X,Atkinson K,Di L.Novel cytochrome P450 reaction phenotyping for low-clearance compounds using the hepatocyte relay method[J].Drug Metab Dispos,2016,44(3):460-465.
[14] Peng CC,Doshi U,Prakash C,et al.A novel plated hepatocyte relay assay (PHRA) for in vitro evaluation of hepatic metabolic clearance of slowly metabolized compounds[J].Drug Metab Lett,2016,10(1):3-15.
[15] Bonn B,Svanberg P,Janefeldt A,et al.Determination of human hepatocyte intrinsic clearance for slowly metabolized compounds:comparison of a primary hepatocyte/stromal cell co-culture with plated primary hepatocytes and HepaRG[J].Drug Metab Dispos,2016,44(4):527-533.
[16] Griffin SJ,Houston JB.Prediction of in vitro intrinsic clearance from hepatocytes:comparison of suspensions and monolayer cultures[J].Drug Metab Dispos,2005,33(1):115-120.
[17] Blanchard N,Alexandre E,Abadie C,et al.Comparison of clearance predictions using primary cultures and suspensions of human hepatocytes[J].Xenobiotica,2005,35(1):1-15.
[18] Smith CM,Nolan CK,Edwards MA,et al.A comprehensive evaluation of metabolic activity and intrinsic clearance in suspensions and monolayer cultures of cryopreserved primary human hepatocytes[J].J Pharm Sci,2012,101(10):3989-4002.
[19] Hoffmaster KA,Turncliff RZ,LeCluyse EL,et al.P-glycoprotein expression,localization,and function in sandwich-cultured primary rat and human hepatocytes:relevance to the hepatobiliary disposition of a model opioid peptide[J].Pharm Res,2004,21(7):1294-1302.
[20] Kanda K,Takahashi R,Yoshikado T,et al.Total hepatocellular disposition profiling of rosuvastatin and pitavastatin in sandwich-cultured human hepatocytes[J].Drug Metab Pharmacokinet,2018,33(3):164-172.
[21] Swift B,Pfeifer ND,Brouwer KL.Sandwich-cultured hepatocytes:an in vitro model to evaluate hepatobiliary transporter-based drug interactions and hepatotoxicity[J].Drug Metab Rev,2010,42(3):446-471.
[22] Brown JH,Das P,DiVito MD,et al.Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro[J].Acta Biomater,2018,73(4):217-227.
[23] Oorts M,Keemink J,Deferm N,et al.Extra collagen overlay prolongs the differentiated phenotype in sandwich-cultured rat hepatocytes[J].J Pharmacol Toxicol Methods,2018,90(6):31-38.
[24] Keemink J,Oorts M,Annaert P.Primary hepatocytes in Sandwich culture[J].Methods Mol Biol,2015,1250(7):175-188.
[25] Lancett P,Williamson B,Barton P,et al.Development and characterization of a human hepatocyte low intrinsic clearance assay for use in drug discovery[J].Drug Metab Dispos,2018,46(8):1169-1178.
[26] Treijtel N,Barendregt A,Freidig AP,et al.Modeling the in vitro intrinsic clearence of the slowly metabolized compound tolbutamide determined in sandwich-cultures rat hepatocytes[J].Drug Metab Dispos,2004,32:884-891.
[27] Liu Y,Wei J,Lu J,et al.Micropatterned coculture of hepatocytes on electrospun fibers as a potential in vitro model for predictive drug metabolism[J].Mater Sci Eng C Mater Biol Appl,2016,63:475-484.
[28] Oda H,Yoshida Y,Kawamura A,et al.Cell shape,cell-cell contact,cell-extracellular matrix contact and cell polarity are all required for the maximum induction of CYP2B1 and CYP2B2 gene expression by phenobarbital in adult rat cultured hepatocytes[J].Biochem Pharmacol,2008,75(5):1209-1217.
[29] Xiao W,Perry G,Komori K,et al.New physiologically-relevant liver tissue model based on hierarchically cocultured primary rat hepatocytes with liver endothelial cells[J].Integr Biol (Camb),2015,7(11):1412-1422.
[30] Kostadinova R,Boess F,Applegate D,et al.A long-term three dimensional liver co-culture system for improved prediction of clinically relevant drug-induced hepatotoxicity[J].Toxicol Appl Pharmacol,2013,268(1):1-16.
[31] Khetani SR,Bhatia SN.Microscale culture of human liver cells for drug development[J].Nat Biotechnol,2008,26(1):120-126.
[32] March S,Ramanan V,Trehan K,et al.Micropatterned coculture of primary human hepatocytes and supportive cells for the study of hepatotropic pathogens[J].Nat Protoc,2015,10(12):2027-2053.
[33] Chan TS,Yu H,Moore A,et al.Meeting the challenge of predicting hepatic clearance of compounds slowly metabolized by cytochrome P450 using a novel hepatocyte model,HepatoPac[J].Drug Metab Dispos,2013,41(12):2024-2032.
[34] Liu YW,Li HN,Yan SL,et al.Hepatocyte cocultures with endothelial cells and fibroblasts on micropatterned fibrous mats to promote liver-specific functions and capillary formation capabilities[J].Biomacromolecules,2014,15(3):1044-1054.
[35] Ukairo O,Kanchagar C,Moore A,et al.Long-term stability of primary rat hepatocytes in micropatterned cocultures[J].J Biochem Mol Toxicol,2013,27(3):204-212.
[36] Shen C,Xu XM,Meng Q,et al.Studies on rifampicin and isoniazid-induced hepatotoxicity[J].J China Pharm Univ(中国药科大学学报),2005,36(3):250-230.
[37] Wang WW,Khetani SR,Krzyzewski S,et al.Assessment of a micropatterned hepatocyte coculture system to generate major human excretory and circulating drug metabolites[J].Drug Metab Dispos,2010,38(10):1900-1905.
[38] Ramsden D,Tweedie DJ,Chan TS,et al.Bridging in vitro and in vivo metabolism and transport of faldaprevir in human using a novel cocultured human hepatocyte system,HepatoPac[J].Drug Metab Dispos,2014,42(3):394-406.
[39] Ramsden D,Tweedie DJ,St George R,et al.Generating an in vitro-in vivo correlation for metabolism and liver enrichment of a hepatitis C virus drug,faldaprevir,using a rat hepatocyte model (HepatoPac)[J].Drug Metab Dispos,2014,42(3):407-414.
[40] Hutzler JM,Ring BJ,Anderson SR.Low-turnover drug molecules:a current challenge for drug metabolism scientists[J].Drug Metab Dispos,2015,43(12):1917-1928.
[41] Chao P,Maguire T,Novik E,et al.Evaluation of a microfluidic based cell culture platform with primary human hepatocytes for the prediction of hepatic clearance in human[J].Biochem Pharmacol,2009,78(6):625-632.
[42] Novik E,Maguire TJ,Chao P,et al.A microfluidic hepatic coculture platform for cell-based drug metabolism studies[J].Biochem Pharmacol,2010,79(7):1036-1044.
[43] Hultman I,Vedin C,Abrahamsson A,et al.Use of HμREL human coculture system for prediction of intrinsic clearance and metabolite formation for slowly metabolized compounds[J].Mol Pharm,2016,13(8):2796-2807.
[44] Dash A,Inman W,Hoffmaster K,et al.Liver tissue engineering in the evaluation of drug safety[J].Exp Opin Drug Metab Toxicol,2009,5(10):1159-1174.
[45] Vivares A,Salle-Lefort S,Arabeyre-Fabre C,et al.Morphological behaviour and metabolic capacity of cryopreserved human primary hepatocytes cultivated in a perfused multiwell device[J].Xenobiotica,2015,45(1):29-44.
[46] Darnell M,Ulvestad M,Ellis E,et al.In vitro evaluation of major in vivo drug metabolic pathways using primary human hepatocytes and HepaRG cells in suspension and a dynamic three-dimensional bioreactor system[J].J Pharmacol Exp Ther,2012,343(1):134-144.
[47] Darnell M,Schreiter T,Zeilinger K,et al.Cytochrome P450-dependent metabolism in HepaRG cells cultured in a dynamic three-dimensional bioreactor[J].Drug Metab Dispos,2011,39(7):1131-1138.
[48] Antherieu S,Chesne C,Li R,et al.Stable expression,activity,and inducibility of cytochromes P450 in differentiated HepaRG cells[J].Drug Metab Dispos,2010,38(3):516-525.
[49] Davies B,Morris T.Physiological parameters in laboratory animals and humans[J].Pharm Res,1993,10(7):1093-1095.
[50] U.S.Food and Drug Administration.Guidance for industry:Safety Testing of Drug Metabolites[EB/OL].(2016-11-22)[2018-12-17].https://www.fda.gov/Drugs/Guidanc- eComplianceRegulatoryInfo-rmation/Guidances/ucm065014.htm.
[51] European Medicines Agency.Guideline on the investigation of drug interactions[EB/OL].(2012-06-21)[2018-12-17]https://www.ema.europa.eu/en/search/search?search_api_views_fulltext=Guideline%-20on%20the%20investigation%20of% 20drug%20interactions.
[52] Matthew Hutzler J,Linder CD,Melton RJ,et al.In vitro-in vivo correlation and translation to the clinical outcome for CJ-13,610,a novel inhibitor of 5-lipoxygenase[J].Drug Metab Dispos,2010,38(7):1113-1121.
[53] Zientek MA,Youdim K.Reaction phenotyping:advances in the experimental strategies used to characterize the contribution of drug-metabolizing enzymes[J].Drug Metab Dispos,2015,43(1):163-181.
[54] Oda S,Fukami T,Yokoi T,et al.A comprehensive review of UDP-glucuronosyltransferase and esterases for drug development[J].Drug Metab Pharmacokinet,2015,30(1):30-51.