Abstract
Two dedicated receptors for bile acids (BAs) have been identified, the nuclear hormone receptor farnesoid X receptor (FXR) and the G protein-coupled receptor TGR5, which represent attractive targets for the treatment of metabolic and chronic liver diseases. Previous work characterized 6α-ethyl-3α,7α-dihydroxy-5β-cholan-24-oic acid (INT-747), a potent and selective FXR agonist, as well as 6α-ethyl-23(S)-methyl-3α,7α,12α-trihydroxy-5β-cholan-24-oic acid (INT-777), a potent and selective TGR5 agonist. Here we characterize 6α-ethyl-3α,7α,23-trihydroxy-24-nor-5β-cholan-23-sulfate sodium salt (INT-767), a novel semisynthetic 23-sulfate derivative of INT-747. INT-767 is a potent agonist for both FXR (mean EC50, 30 nM by PerkinElmer AlphaScreen assay) and TGR5 (mean EC50, 630 nM by time resolved-fluorescence resonance energy transfer), the first compound described so far to potently and selectively activate both BA receptors. INT-767 does not show cytotoxic effects in HepG2 cells, does not inhibit cytochrome P450 enzymes, is highly stable to phase I and II enzymatic modifications, and does not inhibit the human ether-a-go-go-related gene potassium channel. In line with its dual activity, INT-767 induces FXR-dependent lipid uptake by adipocytes, with the beneficial effect of shuttling lipids from central hepatic to peripheral fat storage, and promotes TGR5-dependent glucagon-like peptide-1 secretion by enteroendocrine cells, a validated target in the treatment of type 2 diabetes. Moreover, INT-767 treatment markedly decreases cholesterol and triglyceride levels in diabetic db/db mice and in mice rendered diabetic by streptozotocin administration. Collectively, these preclinical results indicate that INT-767 is a safe and effective modulator of FXR and TGR5-dependent pathways, suggesting potential clinical applications in the treatment of liver and metabolic diseases.
Footnotes
↵ The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.
This work was funded in part by the National Institutes of Health National Institute of Diabetes and Digestive and Kidney Diseases [Grant U01-DK076134] and the National Institutes of Health National Institute on Aging [Grant R01-AG026529] (both to M.L., X.W., T.J.).
Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.
doi:10.1124/mol.110.064501.
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ABBREVIATIONS:
- BA
- bile acid
- FXR
- farnesoid X receptor
- GPCR
- G protein-coupled receptor
- LCA
- lithocholic acid
- CRE
- cAMP response element
- GLP-1
- glucagon-like peptide-1
- INT-747
- 6α-ethyl-3α,7α-dihydroxy-5β-cholan-24-oic acid
- CDCA
- chenodeoxycholic acid
- INT-777
- 6α-ethyl-23(S)-methyl-3α,7α,12α-trihydroxy-5β-cholan-24-oic acid
- INT-767
- 6α-ethyl-3α,7α,23-trihydroxy-24-nor-5β-cholan-23-sulfate sodium salt
- DMSO
- dimethyl sulfoxide
- HEK
- human embryonic kidney
- FCS
- fetal calf serum
- DMEM
- Dulbecco's modified Eagle's medium high-glucose
- FXRE
- FXR-responsive element
- PCR
- polymerase chain reaction
- SR-12813
- [[3,5-bis(1,1-dimethylethyl)-4-hydroxyphenyl]ethenylidene]bis-phosphonic acid tetraethyl ester
- T0901317
- N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]-benzenesulfonamide
- PPAR
- peroxisome proliferator-activated receptor
- GW7647
- 2-methyl-2-[[4-[2-[[(cyclohexylamino)carbonyl](4-cyclohexylbutyl)amino]ethyl] phenyl]thio]-propanoic acid
- GE1929
- N-(2-benzoylphenyl)-O-[2-(methyl-2-pyridinylamino)ethyl]-l-tyrosine hydrochloride
- CAR
- constitutive androstane receptor
- D2
- type 2 iodothyronine deiodinase
- LDH
- lactate dehydrogenase
- BrdU
- bromodeoxyuridine
- CYP450
- cytochrome P450
- MS
- mass spectrometry
- hERG
- ether-a-go-go related gene
- E-4031
- N-[4-[[1-[2-(6-methyl-2-pyridinyl)ethyl]-4-piperidinyl]carbonyl]phenyl]methanesulfonamide dihydrochloride
- mP
- millipolarization
- ORO
- Oil Red O
- siRNA
- small interfering RNA
- WT
- wild-type
- STZ
- streptozotocin
- WD
- Western diet
- LDL
- low-density lipoprotein
- HDL
- high-density lipoprotein
- h
- human
- TR-FRET
- time resolved-fluorescence resonance energy transfer
- NT
- no treatment
- SR-BI
- scavenger receptor type I B1.
- Received March 6, 2010.
- Accepted July 14, 2010.
- Copyright © 2010 The American Society for Pharmacology and Experimental Therapeutics
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