Use of HμREL Human Coculture System for Prediction of Intrinsic Clearance and Metabolite Formation for Slowly Metabolized Compounds

Hultman, Ia; Vedin, Charlotta; Abrahamsson, Anna; Winiwarter, Susanne; Darnell, Malin

doi:10.1021/acs.molpharmaceut.6b00396

Cited by 51 publications

(33 citation statements)

References 34 publications

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“…All infections were carried out between 7–10 days following plating at a point previously determined as optimal for restoring hepatic functions in the cultures 29, 32, 33 . To determine whether the time point of initial HBV infection could possibly be extended further, we challenged mixed donor SACC-PHHs with HBVcc 10, 15, or 20 days post-plating (schematic Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

Long-term hepatitis B infection in a scalable hepatic co-culture system

et al. 2017

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Hepatitis B virus causes chronic infections in 250 million people worldwide. Chronic hepatitis B virus carriers are at risk of developing fibrosis, cirrhosis, and hepatocellular carcinoma. A prophylactic vaccine exists and currently available antivirals can suppress but rarely cure chronic infections. The study of hepatitis B virus and development of curative antivirals are hampered by a scarcity of models that mimic infection in a physiologically relevant, cellular context. Here, we show that cell-culture and patient-derived hepatitis B virus can establish persistent infection for over 30 days in a self-assembling, primary hepatocyte co-culture system. Importantly, infection can be established without antiviral immune suppression, and susceptibility is not donor dependent. The platform is scalable to microwell formats, and we provide proof-of-concept for its use in testing entry inhibitors and antiviral compounds.

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Section: Resultsmentioning

confidence: 99%

Long-term hepatitis B infection in a scalable hepatic co-culture system

et al. 2017

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“…They reported that CL int, in vitro estimated using the relay method showed excellent predictions of human CL int, in vivo . Further, novel in vitro systems such as HepatoPac 44,45) and HμREL, 46,47) which are micropatterned hepatocyte coculture systems, and cell systems such as upcyte, 48) which is derived from human hepatocytes, have been used to estimate low CL int, in vitro and predict in vivo CL in humans.…”

Section: Prediction Of Human Pk Using Ivivementioning

confidence: 99%

Utility of Chimeric Mice with Humanized Liver for Predicting Human Pharmacokinetics in Drug Discovery: Comparison with in Vitro–in Vivo Extrapolation and Allometric Scaling

Naritomi

Sanoh

Ohta

2019

Biological & Pharmaceutical Bulletin

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Predicting human pharmacokinetics (PK) such as clearance (CL) and volume of distribution (Vd) is a critical component of drug discovery. These predictions are mainly performed by in vitro-in vivo extrapolation (IVIVE) using human biological samples, such as hepatic microsomes and hepatocytes. However, some issues with this process have arisen, such as inconsistencies between in vitro and in vivo findings; the integration of predicted CYP, non-CYP and transporter-mediated human PK; and the difficulty of evaluating very metabolically stable compounds. Various approaches to solving these issues have been reported. Allometric scaling using experimental animals has also often been used. However, this method has also shown many problems due to interspecies differences, albeit that various correction methods have been proposed. Another approach involves the production of chimeric mice with humanized liver via the transplantation of human hepatocytes into mice. The livers of these mice are repopulated mostly with human hepatocytes and express human drug-metabolizing enzymes and drug transporters, suggesting that these mice are useful for solving the issues of IVIVE and allometric scaling, and more reliably predicting human PK. In this review, we summarize human PK prediction methods using IVIVE, allometric scaling and chimeric mice with humanized liver, and discuss the utility of predicting human PK in drug discovery by comparing these chimeric mice with IVIVE and allometric scaling.

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“…The physiological human scaling factors were 120 million cells/g liver, 25.7 g liver/kg body weight and 21 mL/min/kg of liver blood flow (Q h ). Fraction unbound in plasma ( f u,p ) and the blood to plasma ratio (R b ) were obtained from the literature, 4,[15][16][17][18] and each fraction unbound in blood ( f b ) was calculated by dividing the value of f u,p by that of R b . The protein binding rate in metabolic culture medium or adherent cells ( f u,inc ) was assumed to be 1.…”

Section: Chemicals and Reagentsmentioning

confidence: 99%

“…However, when hepatocytes are used in suspension assay, metabolic activity or hepatic function are drastically reduced within several hours. Further, the number of slowly metabolized drug candidates, which are difficult to evaluate in conventional assay systems, are increasing in the field of drug discovery, thus, evaluation methods for low CL drug candidates such as the Relay method 3) and co-culture system 4,5) have been developed. In addition, human PK prediction using chimeric mice has attracted attention, and their use in drug discovery has increased.…”

Section: Introductionmentioning

confidence: 99%

Prediction of Human Hepatic Clearance for Cytochrome P450 Substrates via a New Culture Method Using the Collagen Vitrigel Membrane Chamber and Fresh Hepatocytes Isolated from Liver Humanized Mice

Watari

Kakiki

Yamasaki

et al. 2019

Biological & Pharmaceutical Bulletin

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In drug discovery, hepatocytes have been widely utilized as in vitro tools for predicting the in vivo hepatic clearance (CL) of drug candidates. However, conventional hepatocyte models do not always reproduce in vivo physiological function, and CYP activities in particular decrease quite rapidly during culture. Furthermore, conventional in vitro assays have limitations in their ability to predict hepatic CL of metabolically stable drug candidates. In order to accurately predict hepatic CL of candidate drugs, a new method of culturing hepatocytes that activates their functional properties, including CYP activities, is in high demand.In the previous study, we established a novel long-term culture method for PXB-cells ® using a collagen vitrigel membrane (CVM) chamber, which can maintain CYP activity and liver specific functions at high levels for several weeks. In this study, the vitrigel culture method was applied to predictions of hepatic CL for 22 CYP typical substrates with low to middle CL, and the prediction accuracy by this method was assessed by comparing CL data between predicted (in vitro intrinsic CL using the dispersion model) and observed (in vivo clinical data) values. The results of this study showed that in vitro CL values for approximately 60% (13/22) and 80% (18/22) of the compounds were predicted within a 2-and 3-fold difference with in vivo CL, respectively. These results suggest that the new culture method using the CVM chamber and PXB-cells is a promising in vitro system for predicting human hepatic CL with high accuracy for CYP substrates, including metabolically stable drug candidates. Key words collagen vitrigel membrane (CVM); hepatocyte; PXB-cell; CYP; in vitro in vivo correlation (IVIVC)

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Use of HμREL Human Coculture System for Prediction of Intrinsic Clearance and Metabolite Formation for Slowly Metabolized Compounds

Cited by 51 publications

References 34 publications

Long-term hepatitis B infection in a scalable hepatic co-culture system

Long-term hepatitis B infection in a scalable hepatic co-culture system

Utility of Chimeric Mice with Humanized Liver for Predicting Human Pharmacokinetics in Drug Discovery: Comparison with <i>in Vitro</i>–<i>in Vivo</i> Extrapolation and Allometric Scaling

Prediction of Human Hepatic Clearance for Cytochrome P450 Substrates <i>via</i> a New Culture Method Using the Collagen Vitrigel Membrane Chamber and Fresh Hepatocytes Isolated from Liver Humanized Mice

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