Method for modulating process mediated by farnesoid activated receptors
AssigneesThe Salk Institute for Biological Studies, Ligand Pharmaceuticals Incorporated, The Government of the United States of America
InventorsRonald M. Evans, Barry M. Forman, Cary A. Weinberger
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Abstract
Farnesyl pyrophosphate, the metabolically active form of farnesol, is a key precursor in the synthesis of cholesterol, carotenoids, steroid hormones, bile acids and other molecules involved in cellular growth and metabolism. A nuclear receptor has been identified that is transcriptionally activated by farnesol and related molecules. This novel signaling pathway can be modulated by the use of key metabolic intermediates (or analogs and/or derivatives thereof) as transcriptional regulatory signals.
Claims24
A method of testing a compound for its ability to activate a farnesoid-activated receptor (FAR) polypeptide,
- wherein said receptor polypeptide is a nuclear receptor and is responsive to the presence of farnesoid to regulate the transcription of genes having response elements organized as an IR1, said method comprising assaying for a reporter protein when cells containing said receptor polypeptide and a reporter construct are contacted with said compound,
- wherein said reporter construct comprises
- a) a promoter that is operable in said cell,
- b) a response element organized as an IR1,
- c) a DNA encoding a reporter protein,
- wherein said DNA is operatively linked to said promoter for transcription of said DNA and said promoter is operatively linked to said response element for activation thereof;
- and
- wherein said activation of said receptor polypeptide results in transcription and expression of the reporter protein.
A method of testing a compound for its ability to activate a famesoid-activated receptor (FAR) polypeptide,
- wherein said receptor polypeptide is a nuclear receptor and is responsive to the presence of farnesoid to regulate the transcription of genes having response elements organized as an IR1,
- said method comprising assaying for a reporter protein when cells containing a chimeric receptor polypeptide comprising the ligand binding domain of said receptor polypeptide and a heterologous DNA binding domain, a reporter construct, and optionally a retinoid X receptor, are contacted with said compound,
- wherein said reporter construct comprises
- a) a promoter that is operable in said cell,
- b) a response element recognizable by the DNA binding domain of said chimeric receptor polypeptide,
- c) a DNA encoding a reporter protein,
- wherein said DNA is operatively linked to said promoter for transcription of said DNA and said promoter is operatively linked to said response element for activation thereof;
- and
- wherein activation of said chimeric receptor polypeptide results in transcription and expression of the reporter protein, thereby identifying a compound that activates said receptor polypeptide.
A method of testing a compound for its ability to inhibit a farnesoid-activated receptor (FAR) polypeptide,
- wherein said receptor polypeptide is a nuclear receptor and is responsive to the presence of farnesoid to regulate the transcription of genes having response elements organized as an IR1, said method comprising contacting cells which contain said receptor polypeptide, a reporter construct, and optionally a retinoid X receptor (RXR), with at least one FAR agonist and optionally a RXR agonist, in the absence or presence of said compound,
- wherein said reporter construct comprises
- a) a promoter that is operable in said cell,
- b) a response element organized as an IR1,
- c) a DNA encoding a reporter protein,
- wherein said DNA is operatively linked to said promoter for transcription of said DNA and said promoter is operatively linked to said response element for activation thereof;
- and
- wherein said inhibition of said receptor polypeptide in the presence of said compound results in reduced transcription and expression of the reporter protein relative to that observed in the absence of said compound.
A method of testing a compound for its ability to inhibit a farnesoid-activated receptor (FAR) polypeptide,
- wherein said receptor polypeptide is a nuclear receptor and is responsive to the presence of farnesoid to regulate the transcription of genes having response elements organized as an IR1,
- said method comprising contacting cells containing a chimeric receptor polypeptide comprising the ligand binding domain of said receptor polypeptide and a heterologous DNA binding domain, a reporter construct, and optionally a retinoid X receptor (RXR), with at least one FAR agonist and optionally a RXR agonist, in the absence or presence of said compound,
- wherein said reporter construct comprises
- a) a promoter that is operable in said cell,
- b) a response element recognizable by the DNA binding domain of said chimeric receptor polypeptide,
- c) a DNA encoding a reporter protein,
- wherein said DNA is operatively linked to said promoter for transcription of said DNA and said promoter is operatively linked to said response element for activation thereof;
- and
- wherein inhibition of said chimeric receptor polypeptide in the presence of said compound results in reduced transcription and expression of the reporter protein relative to that observed in the absence of said compound, thereby identifying a compound that inhibits said receptor polypeptide.
Dependencies are inferred from the claim text.