The central goal of this R01 is to focus on explicitly defining novel antagonist binding pharmacophore on Pregnane X Receptor (PXR). In doing so, additional goals include development of non-toxic azole antagonists that would serve to chemically probe PXR activity and phenotype(s) in different tissues. In silico modeling parameters will continuously be improved as we obtain potent and specific PXR inhibitors. These models could then guide the development of novel small molecule antagonists originating from different chemical entities. The long-term goal is to eventually develop non-toxic antagonists of PXR that can be used as clinical modulators of cancer cell proliferation and drug resistance (e.g., PXR activation induces cancer cell proliferation and drug resistance). It is also hoped that these antagonists will enhance the activity, and minimize the toxicity, of select antineoplastic agents (e.g., tamoxifen, paclitaxel are PXR agonist at concentrations observed at steady-state in humans). Towards this end, we have identified and characterized two novel PXR antagonists, ketoconazole and coumestrol, that specifically disrupt the function of activated (ligand-bound) PXR. In subsequent studies, we have shown that ketoconazole: (i) binds to receptor and disrupts coregulator-receptor interactions in activated PXR;(2) does not displace activating drugs from the ligand-binding pocket of PXR;(iii) retained antagonism of mutant forms of PXR containing ligand-binding pocket filling mutants;and (iv) is unable to antagonize mutant forms of PXR containing alterations in the surface coregulator AF-2 binding site. Thus, we have formulated a model for PXR antagonism in which disruption of function is mediated either by allosteric modification of the receptor or by competition with coregulator binding. We now propose to evaluate this model using structural, molecular, biochemical, and genetic systems to characterize the mechanism by which PXR-directed antagonist ketoconazole and related compounds inhibit receptor activation.

Public Health Relevance

Adopted orphan nuclear receptor, Pregnane X Receptor (PXR), plays a mechanistic role in drug-drug interactions and cancer drug resistance. These functions of the receptor are newly discovered and since this receptor has organ-specific functions, many of its roles in other tissues (e.g., gut) are unknown. PXR is activated by agonists (ligands) and inactivated by antagonists. Our laboratory and co-investigators have taken a lead role in defining novel antagonists of PXR that bind to a region outside the ligand binding pocket. We believe that these antagonists will be necessary to probe PXR function and define all the phenotypes of PXR in mice and humans. In this proposal, we will evaluate our model of antagonism using structural, molecular, biochemical, and genetic systems. These studies will be vital for the generation on non-toxic compounds that antagonize PXR function - a tool necessary to probe PXR function in cells.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
Project #
Application #
Study Section
Drug Discovery and Molecular Pharmacology Study Section (DMP)
Program Officer
Lees, Robert G
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Albert Einstein College of Medicine
Internal Medicine/Medicine
Schools of Medicine
United States
Zip Code
Stepankova, Martina; Bartonkova, Iveta; Jiskrova, Eva et al. (2018) Methylindoles and Methoxyindoles are Agonists and Antagonists of Human Aryl Hydrocarbon Receptor. Mol Pharmacol 93:631-644
Hudson, Grace M; Flannigan, Kyle L; Erickson, Sarah L et al. (2017) Constitutive androstane receptor regulates the intestinal mucosal response to injury. Br J Pharmacol 174:1857-1871
Garg, Aditya; Zhao, Angela; Erickson, Sarah L et al. (2016) Pregnane X Receptor Activation Attenuates Inflammation-Associated Intestinal Epithelial Barrier Dysfunction by Inhibiting Cytokine-Induced Myosin Light-Chain Kinase Expression and c-Jun N-Terminal Kinase 1/2 Activation. J Pharmacol Exp Ther 359:91-101
Pasquel, Danielle; Doricakova, Aneta; Li, Hao et al. (2016) Acetylation of lysine 109 modulates pregnane X receptor DNA binding and transcriptional activity. Biochim Biophys Acta 1859:1155-1169
Ranhotra, Harmit S; Flannigan, Kyle L; Brave, Martina et al. (2016) Xenobiotic Receptor-Mediated Regulation of Intestinal Barrier Function and Innate Immunity. Nucl Receptor Res 3:
Pondugula, Satyanarayana R; Pavek, Petr; Mani, Sridhar (2016) Pregnane X Receptor and Cancer: Context-Specificity is Key. Nucl Receptor Res 3:
Zhang, Jingjing; Ding, Lili; Wang, Baocan et al. (2015) Notoginsenoside R1 attenuates experimental inflammatory bowel disease via pregnane X receptor activation. J Pharmacol Exp Ther 352:315-24
Pondugula, Satyanarayana R; Flannery, Patrick C; Abbott, Kodye L et al. (2015) Diindolylmethane, a naturally occurring compound, induces CYP3A4 and MDR1 gene expression by activating human PXR. Toxicol Lett 232:580-9
Zhang, Jingjing; Dou, Wei; Zhang, Eryun et al. (2014) Paeoniflorin abrogates DSS-induced colitis via a TLR4-dependent pathway. Am J Physiol Gastrointest Liver Physiol 306:G27-36
Venkatesh, Madhukumar; Mukherjee, Subhajit; Wang, Hongwei et al. (2014) Symbiotic bacterial metabolites regulate gastrointestinal barrier function via the xenobiotic sensor PXR and Toll-like receptor 4. Immunity 41:296-310

Showing the most recent 10 out of 32 publications