Constitutive androstane receptor (CAR) and pregnane X receptor (PXR) are closely related nuclear receptors with overlapping regulatory functions in xenobiotic clearance but distinct roles in endobiotic metabolism. Car activation has been demonstrated to ameliorate hypercholesterolemia by regulating cholesterol metabolism and bile acid elimination, whereas PXR activation is associated with hypercholesterolemia and liver steatosis. Here we show a human CAR agonist/PXR antagonist, MI-883, which effectively regulates genes related to xenobiotic metabolism and cholesterol/bile acid homeostasis by leveraging CAR and PXR interactions in gene regulation. Through comprehensive analyses utilizing lipidomics, bile acid metabolomics, and transcriptomics in humanized PXR-CAR-CYP3A4/3A7 mice fed high-fat and high-cholesterol diets, we demonstrate that MI-883 significantly reduces plasma cholesterol levels and enhances fecal bile acid excretion. This work paves the way for the development of ligands targeting multiple xenobiotic nuclear receptors. Such ligands hold the potential for precise modulation of liver metabolism, offering new therapeutic strategies for metabolic disorders.

1st Faculty of Medicine Charles University Prague Czech Republic

4th Department of Internal Medicine General University Hospital Prague and 1st Faculty of Medicine Charles University Prague Czech Republic

Czech Centre for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences Prague Czech Republic

Department of Analytical Chemistry University of Pardubice Faculty of Chemical Technology Pardubice Czech Republic

Department of Biochemistry Faculty of Medicine in Hradec Králové Charles University Hradec Králové Czech Republic

Department of Pharmacology and Toxicology Faculty of Pharmacy in Hradec Králové Charles University Hradec Králové Czech Republic

Dr Margarete Fischer Bosch Institute of Clinical Pharmacology Stuttgart and University of Tuebingen Tuebingen Germany

Institute of Medical Biochemistry and Laboratory Diagnostics General University Hospital Prague and 1st Faculty of Medicine Charles University Prague Czech Republic

Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences Prague Czech Republic

Institute of Pharmacology Faculty of Medicine in Hradec Králové Charles University Hradec Králové Czech Republic

Institute of Pharmacy Pharmaceutical Medicinal Chemistry and Tübingen Center for Academic Drug Discovery Eberhard Karls University Tübingen Tübingen Germany

Military Faculty of Medicine University of Defence Hradec Králové Czech Republic

PamGene 's Hertogenbosch The Netherlands

School of Pharmacy Faculty of Health Sciences University of Eastern Finland Kuopio Finland

Section of Pharmacogenetics Department of Physiology and Pharmacology Karolinska Institutet Stockholm Sweden

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