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Identification of Novel Activators of Constitutive Androstane Receptor from FDA-Approved Drugs by Integrated Computational and Biological Approaches

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Abstract

Purpose

The constitutive androstane receptor (CAR, NR1I3) is a xenobiotic sensor governing the transcription of numerous hepatic genes associated with drug metabolism and clearance. Recent evidence suggests that CAR also modulates energy homeostasis and cancer development. Thus, identification of novel human (h) CAR activators is of both clinical importance and scientific interest.

Methods

Docking and ligand-based structure-activity models were used for virtual screening of a database containing over 2000 FDA-approved drugs. Identified lead compounds were evaluated in cell-based reporter assays to determine hCAR activation. Potential activators were further tested in human primary hepatocytes (HPHs) for the expression of the prototypical hCAR target gene CYP2B6.

Results

Nineteen lead compounds with optimal modeling parameters were selected for biological evaluation. Seven of the 19 leads exhibited moderate to potent activation of hCAR. Five out of the seven compounds translocated hCAR from the cytoplasm to the nucleus of HPHs in a concentration-dependent manner. These compounds also induce the expression of CYP2B6 in HPHs with rank-order of efficacies closely resembling that of hCAR activation.

Conclusion

These results indicate that our strategically integrated approaches are effective in the identification of novel hCAR modulators, which may function as valuable research tools or potential therapeutic molecules.

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Abbreviations

Ad/EYFP-hCAR:

adenovirus expressing enhanced yellow fluorescent protein-tagged human CAR

AMI:

amiodarone

CAL:

carvedilol

CAR:

constitutive androstane receptor

CITCO:

6-(4-chlorophenyl) imidazo[2,1-b][1,3]-thiazole-5-carbaldehyde-O-(3,4-dichlorobenzyl)oxime

CYP:

cytochrome P450

DMSO:

dimethyl sulfoxide

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

HPH:

human primary hepatocytes

LBD:

ligand-binding domain

NEF:

nefazodone

NIC:

nicardipine

OCT:

octicizer

PB:

phenobarbital

PXR:

pregnane X receptor

RT-PCR:

reverse transcription-polymerase chain reaction

TEL:

telmisartan

TOL:

tolnaftate

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Acknowledgments and Disclosures

The authors thank Dr. James Polli (The University of Maryland School of Pharmacy) for kindly offering multiple compounds and Dr. Alex MacKerell (The University of Maryland School of Pharmacy) for making the Discovery Studio available for this study. We also thank Dr. Sean Ekins (Collaborations in Chemistry, Jenkintown, PA) for offering initial CDD database and Dr. Taiji Oashi, a previous lab member from Dr. MacKerell’s lab for database optimization. The authors appreciatively acknowledge The University of Maryland Medical Center and Life Technologies (Durham, NC) for providing the human hepatocytes used in this study. The research is supported in part by the National Institutes of Health Grants DK061652 (H.W) and DK061425 (P.S).

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 Penn. Street, Baltimore, Maryland, 21201, USA

    Caitlin Lynch, Yongmei Pan, Linhao Li, Peter W. Swaan & Hongbing Wang

  2. Life Technologies Corporations, Durham, North Carolina, 27707, USA

    Stephen S. Ferguson

  3. NIH Chemical Genomics Center, National Institutes of Health, Bethesda, Maryland, 20892, USA

    Menghang Xia

Authors
  1. Caitlin Lynch
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  2. Yongmei Pan
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  3. Linhao Li
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  4. Stephen S. Ferguson
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  5. Menghang Xia
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  6. Peter W. Swaan
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  7. Hongbing Wang
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Corresponding author

Correspondence to Hongbing Wang.

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ESM 1

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Figure S1

Chemical structures of training compounds. Seventeen known human CAR ligands extracted from literature were used as training compounds to generate the common features of pharmacophore model in this study. The chemical structures of these compounds were drawn using ChemDraw Ultra 10. (JPEG 115 kb)

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Figure S2

Chemical structures of tested lead compounds. Nineteen lead compounds with optimal pharmacophore parameters from virtual screening of the FDA-approved drug data base were selected for biological assessment of CAR activation. The chemical structures of these compounds were drawn using ChemDraw Ultra 10. (JPEG 151 kb)

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Lynch, C., Pan, Y., Li, L. et al. Identification of Novel Activators of Constitutive Androstane Receptor from FDA-Approved Drugs by Integrated Computational and Biological Approaches. Pharm Res 30, 489–501 (2013). https://doi.org/10.1007/s11095-012-0895-1

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  • DOI: https://doi.org/10.1007/s11095-012-0895-1

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