Modeling Therapeutic Antibody-Small Molecule Drug-Drug Interactions Using a Three-Dimensional Perfusable Human Liver Coculture Platform.
Traditional in vitro human liver cell culture models lose key hepatic functions such as metabolic activity during short-term culture. Advanced three-dimensional (3D) liver coculture platforms offer the potential for extended hepatocyte functionality and allow for the study of more complex biologic interactions, which can improve and refine human drug safety evaluations. Here, we use a perfusion flow 3D microreactor platform for the coculture of cryopreserved primary human hepatocytes and Kupffer cells to study the regulation of cytochrome P450 3A4 isoform (CYP3A4) activity by chronic interleukin 6 (IL-6)-mediated inflammation over 2 weeks. Hepatocyte cultures remained stable over 2 weeks, with consistent albumin production and basal IL-6 levels. Direct IL-6 stimulation that mimics an inflammatory state induced a dose-dependent suppression of CYP3A4 activity, an increase in C-reactive protein (CRP) secretion, and a decrease in shed soluble interleukin-6 receptor (IL-6R) levels, indicating expected hepatic IL-6 bioactivity. Tocilizumab, an anti-IL-6R monoclonal antibody used to treat rheumatoid arthritis, has been demonstrated clinically to impact small molecule drug pharmacokinetics by modulating cytochrome P450 enzyme activities, an effect not observed in traditional hepatic cultures. We have now recapitulated the clinical observation in a 3D bioreactor system. Tocilizumab was shown to desuppress CYP3A4 activity while reducing the CRP concentration after 72 hours in the continued presence of IL-6. This change in CYP3A4 activity decreased the half-life and area under the curve up to the last measurable concentration (AUClast) of the small molecule CYP3A4 substrate simvastatin hydroxy acid, measured before and after tocilizumab treatment. We conclude that next-generation in vitro liver culture platforms are well suited for these types of long-term treatment studies and show promise for improved drug safety assessment.
Comparative Biology and Safety Science Laboratory, Amgen, Inc., Cambridge, Massachusetts (T.J.L.); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts (T.J.L., L.G.G.); Comparative Biology and Safety Science Laboratory, Amgen, Inc., Thousand Oaks, California (P.A.C., R.T.D., H.H., H.M., C.A.); Center for Gynepathology Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (L.G.G.); Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.); Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.).
Thomas J
TJ
Long
Comparative Biology and Safety Science Laboratory, Amgen, Inc., Cambridge, Massachusetts (T.J.L.); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts (T.J.L., L.G.G.); Comparative Biology and Safety Science Laboratory, Amgen, Inc., Thousand Oaks, California (P.A.C., R.T.D., H.H., H.M., C.A.); Center for Gynepathology Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (L.G.G.); Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.); Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.).
Patrick A
PA
Cosgrove
Comparative Biology and Safety Science Laboratory, Amgen, Inc., Cambridge, Massachusetts (T.J.L.); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts (T.J.L., L.G.G.); Comparative Biology and Safety Science Laboratory, Amgen, Inc., Thousand Oaks, California (P.A.C., R.T.D., H.H., H.M., C.A.); Center for Gynepathology Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (L.G.G.); Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.); Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.).
Robert T
RT
Dunn
Comparative Biology and Safety Science Laboratory, Amgen, Inc., Cambridge, Massachusetts (T.J.L.); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts (T.J.L., L.G.G.); Comparative Biology and Safety Science Laboratory, Amgen, Inc., Thousand Oaks, California (P.A.C., R.T.D., H.H., H.M., C.A.); Center for Gynepathology Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (L.G.G.); Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.); Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.).
Donna B
DB
Stolz
Comparative Biology and Safety Science Laboratory, Amgen, Inc., Cambridge, Massachusetts (T.J.L.); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts (T.J.L., L.G.G.); Comparative Biology and Safety Science Laboratory, Amgen, Inc., Thousand Oaks, California (P.A.C., R.T.D., H.H., H.M., C.A.); Center for Gynepathology Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (L.G.G.); Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.); Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.).
Hisham
H
Hamadeh
Comparative Biology and Safety Science Laboratory, Amgen, Inc., Cambridge, Massachusetts (T.J.L.); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts (T.J.L., L.G.G.); Comparative Biology and Safety Science Laboratory, Amgen, Inc., Thousand Oaks, California (P.A.C., R.T.D., H.H., H.M., C.A.); Center for Gynepathology Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (L.G.G.); Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.); Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.).
Cynthia
C
Afshari
Comparative Biology and Safety Science Laboratory, Amgen, Inc., Cambridge, Massachusetts (T.J.L.); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts (T.J.L., L.G.G.); Comparative Biology and Safety Science Laboratory, Amgen, Inc., Thousand Oaks, California (P.A.C., R.T.D., H.H., H.M., C.A.); Center for Gynepathology Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (L.G.G.); Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.); Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.).
Helen
H
McBride
Comparative Biology and Safety Science Laboratory, Amgen, Inc., Cambridge, Massachusetts (T.J.L.); Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts (T.J.L., L.G.G.); Comparative Biology and Safety Science Laboratory, Amgen, Inc., Thousand Oaks, California (P.A.C., R.T.D., H.H., H.M., C.A.); Center for Gynepathology Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (L.G.G.); Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.); Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (D.B.S.) griff@mit.edu.
Linda G
LG
Griffith
D000911:Antibodies, Monoclonal / Q000378:metabolism*; D061067:Antibodies, Monoclonal, Humanized / Q000378:metabolism; D002097:C-Reactive Protein / Q000378:metabolism; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D004347:Drug Interactions / Q000502:physiology*; D006207:Half-Life; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D007728:Kupffer Cells / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D015850:Interleukin-6; D054852:Small Molecule Libraries; D002097:C-Reactive Protein; D051544:Cytochrome P-450 CYP3A; C502936:tocilizumab