A Semi-Physiologically Based Pharmacokinetic Model Describing the Altered Metabolism of Midazolam Due to Inflammation in Mice
To investigate influence of inflammation on metabolism and pharmacokinetics (PK) of midazolam (MDZ) and construct a semi-physiologically based pharmacokinetic (PBPK) model to predict PK in mice with inflammatory disease.
Glucose-6-phosphate isomerase (GPI)-mediated inflammation was used as a preclinical model of arthritis in DBA/1 mice. CYP3A substrate MDZ was selected to study changes in metabolism and PK during the inflammation. The semi-PBPK model was constructed using mouse physiological parameters, liver microsome metabolism, and healthy animal PK data. In addition, serum cytokine, and liver-CYP (cytochrome P450 enzymes) mRNA levels were examined.
The in vitro metabolite formation rate was suppressed in liver microsomes prepared from the GPI-treated mice as compared to the healthy mice. Further, clearance of MDZ was reduced during inflammation as compared to the healthy group. Finally, the semi-PBPK model was used to predict PK of MDZ after GPI-mediated inflammation. IL-6 and TNF-α levels were elevated and liver-cyp3a11 mRNA was reduced after GPI treatment.
The semi-PBPK model successfully predicted PK parameters of MDZ in the disease state. The model may be applied to predict PK of other drugs under disease conditions using healthy animal PK and liver microsomal data as inputs.
Key wordscytokines drug metabolism glucose-6-phosphate isomerase inflammation physiologically based pharmacokinetic model
Constitutive androstane receptor
Dried blood spot
Hepatic nuclear factor-4α
Human liver microsomes
Liquid Chromatography-Mass Spectrometry
Mouse liver microsomes
New chemical entity
Physiologically based pharmacokinetic
Pregnane X receptor
Quantitative polymerase chain reaction
Severe combined immune deficient
The authors acknowledge Michael Mohutsky for help with the in vitro metabolism work, Tom Kern (Covance Inc.) for conducting in vivo pharmacokinetic experiments, George Searfoss for CYP mRNA measurements, Bridget Morse for suggestions regarding the semi-PBPK model, and Daniel Mudra for critically reading the manuscript and providing suggestions. Eli Lilly provided support for an internship by NV and funded laboratory and animal studies. NV and MLF were partially supported by a grant from NIH (R01CA173292, PI: Forrest) during analysis and development of the model. NV was partially supported by a Higuchi Fellowship and the Department of Pharmaceutical Chemistry, The University of Kansas.
Compliance with Ethical Standards
Conflict of Interest
None to declare.
- 6.Xu Y, Hijazi Y, Wolf A, Wu B, Sun YN, Zhu M. Physiologically based pharmacokinetic model to assess the influence of Blinatumomab-mediated cytokine elevations on cytochrome P450 enzyme activity. CPT: pharmacometrics & systems pharmacology. 2015;4(9):507–15.Google Scholar
- 8.Matsumoto I, Zhang H, Yasukochi T, Iwanami K, Tanaka Y, Inoue A, et al. Therapeutic effects of antibodies to tumor necrosis factor-alpha, interleukin-6 and cytotoxic T-lymphocyte antigen 4 immunoglobulin in mice with glucose-6-phosphate isomerase induced arthritis. Arthritis Res Ther. 2008;10(3):R66.CrossRefPubMedPubMedCentralGoogle Scholar
- 21.Kuze J, Mutoh T, Takenaka T, Morisaki K, Nakura H, Hanioka N, et al. Separate evaluation of intestinal and hepatic metabolism of three benzodiazepines in rats with cannulated portal and jugular veins: comparison with the profile in non-cannulated mice. Xenobiotica. 2009;39(11):871–80.CrossRefPubMedGoogle Scholar
- 27.Kuze J, Mutoh T, Takenaka T, Oda N, Hanioka N, Narimatsu S. Evaluation of animal models for intestinal first-pass metabolism of drug candidates to be metabolized by CYP3A enzymes via in vivo and in vitro oxidation of midazolam and triazolam. Xenobiotica. 2013;43(7):598–606.CrossRefPubMedGoogle Scholar
- 28.Barter ZE, Bayliss MK, Beaune PH, Boobis AR, Carlile DJ, Edwards RJ, et al. Scaling factors for the extrapolation of in vivo metabolic drug clearance from in vitro data: reaching a consensus on values of human micro-somal protein and hepatocellularity per gram of liver. Curr Drug Metab. 2007;8(1):33–45.CrossRefPubMedGoogle Scholar
- 33.Wang J, Xia S, Xue W, Wang D, Sai Y, Liu L, et al. A semi-physiologically-based pharmacokinetic model characterizing mechanism-based auto-inhibition to predict stereoselective pharmacokinetics of verapamil and its metabolite norverapamil in human. Eur J Pharm Sci. 2013;50(3):290–302.CrossRefPubMedGoogle Scholar