Abstract
The availability of high-affinity agonists for peroxisome proliferator-activated receptor-β/δ (PPARβ/δ) has led to significant advances in our understanding of the functional role of PPARβ/δ. In this study, a new PPARβ/δ antagonist, 4-chloro-N-(2-{[5-trifluoromethyl)-2-pyridyl]sulfonyl}ethyl)benzamide (GSK3787), was characterized using in vivo and in vitro models. Orally administered GSK3787 caused antagonism of 4-[2-(3-fluoro-4-trifluoromethyl-phenyl)-4-methyl-thiazol-5-ylmethylsulfanyl]-2-methyl-phenoxy}-acetic acid (GW0742)-induced up-regulation of Angptl4 and Adrp mRNA expression in wild-type mouse colon but not in Pparβ/δ-null mouse colon. Chromatin immunoprecipitation (ChIP) analysis indicates that this correlated with reduced promoter occupancy of PPARβ/δ on the Angptl4 and Adrp genes. Reporter assays demonstrated antagonism of PPARβ/δ activity and weak antagonism and agonism of PPARγ activity but no effect on PPARα activity. Time-resolved fluorescence resonance energy transfer assays confirmed the ability of GSK3787 to modulate the association of both PPARβ/δ and PPARγ coregulator peptides in response to ligand activation, consistent with reporter assays. In vivo and in vitro analysis indicates that the efficacy of GSK3787 to modulate PPARγ activity is markedly lower than the efficacy of GSK3787 to act as a PPARβ/δ antagonist. GSK3787 antagonized GW0742-induced expression of Angptl4 in mouse fibroblasts, mouse keratinocytes, and human cancer cell lines. Cell proliferation was unchanged in response to either GW0742 or GSK3787 in human cancer cell lines. Results from these studies demonstrate that GSK3787 can antagonize PPARβ/δ in vivo, thus providing a new strategy to delineate the functional role of a receptor with great potential as a therapeutic target for the treatment and prevention of disease.
Footnotes
↵
The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.This work was supported by the National Institutes of Health National Cancer Institute [Grants CA124533, CA126826, CA141029]; the Deutsche Forschungsgemeinschaft [Grant SFB-TR17/A3]; and in part by the Intramural Research Program of the National Institutes of Health National Cancer Institute.
Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.
doi:10.1124/mol.110.065508.
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ABBREVIATIONS:
- PPAR
- peroxisome proliferator-activated receptor
- Adrp
- adipocyte differentiation-related protein
- ANOVA
- analysis of variance
- Angptl4
- angiopoietin-like protein 4
- ChIP
- chromatin immunoprecipitation
- DMSO
- dimethyl sulfoxide
- DMEM
- Dulbecco's modified Eagle's medium
- FBS
- fetal bovine serum
- GAPDH
- glyceraldehyde 3-phosphate dehydrogenase
- PPRE
- peroxisome proliferator response element
- PCR
- polymerase chain reaction
- TR-FRET
- time-resolved fluorescence resonance energy transfer
- TRAP220/DRIP-1
- thyroid hormone receptor-associated protein 220/vitamin D receptor interacting protein-1
- SMRT-ID2
- silencing mediator for retinoid and thyroid hormone receptors interaction domain 2
- NCoR-ID2
- nuclear receptor corepressor interaction domain 2
- AcH4
- acetylated histone H4
- LBD
- ligand binding domain
- qPCR
- quantitative real-time polymerase chain reaction
- PDK4
- pyruvate dehydrogenase kinase 4
- CPT1a
- carnitine palmitoyl transferase 1a
- GST
- glutathione transferase
- GSK3787
- 4-chloro-N-(2-{[5-trifluoromethyl)-2-pyridyl]sulfonyl}ethyl)benzamide
- GW0742
- 4-[2-(3-fluoro-4-trifluoromethyl-phenyl)-4-methyl-thiazol-5-ylmethylsulfanyl]-2-methyl-phenoxy}-acetic acid
- GW501516
- (2-methyl-4-(((4-methyl-2-(4-(trifluoromethyl) phenyl)-5-thiazolyl)methyl)thio)phenoxy)-acetic acid
- GSK0660
- 3-[[[2-methoxy-4-(phenylamino)phenyl]amino]sulfonyl]-2-thiophenecarboxylic acid methyl ester
- GW1929
- N-(2-benzoylphenyl)-O-[2-(methyl-2-pyridinylamino)ethyl]-l-tyrosine hydrochloride
- GW7647
- 2-[[4-[2-[[(cyclohexylamino)carbonyl](4-cyclohexylbutyl)amino]ethyl]phenyl]thio]-2-methylpropanoic acid.
- Received April 9, 2010.
- Accepted June 1, 2010.
- U.S. Government work not protected by U.S. copyright
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Cellular and Pharmacological Selectivity of the Peroxisome Proliferator-Activated Receptor-β/δ Antagonist GSK3787
