Constitutive androstane receptor
AssigneeSmithKline Beecham Corporation
InventorsJon L. Collins, Derek J. Parks
Life of this patent
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Abstract
The present invention relates, in general, to nuclear receptors and, in particular, to the Constitutive Androstane Receptor (CAR; NR 1I3) and to a method of identifying ligands therefor.
Claims18
A method of screening a selected compound for its ability to inhibit the binding of clotrimazole to a Constitutive Androstane Receptor (CAR) ligand binding domain (LBD)-containing polypeptide comprising:
- i) contacting said selected compound and clotrimazole with said CAR LBD-containing polypeptide under conditions such that clotrimazole can bind to said CAR LBD-containing polypeptide in the absence of said selected compound, and
- ii) determining the amount of clotrimazole bound to said CAR LBD-containing polypeptide and comparing that amount to an amount of clotrimazole bound to said CAR LBD-containing polypeptide in the absence of said selected compound,
- wherein a reduction in the amount of clotrimazole bound to said CAR LBD-containing polypeptide in the presence of said selected compound indicates that said selected compound inhibits the binding of clotrimazole to said CAR LBD-containing polypeptide.
A method of determining whether a ligand for a selected nuclear receptor is non-specific for the selected nuclear receptor, comprising
- i) contacting a ligand that binds the selected nuclear receptor with clotrimazole and a human Constitutive Androstane Receptor (hCAR) ligand binding domain (LBD)-containing polypeptide under conditions such that clotrimazole can bind to the hCAR LBD-containing polypeptide in the absence of the ligand, and
- ii) determining the amount of clotrimazole bound to the hCAR LBD-containing polypeptide,
- wherein an amount of clotrimazole bound to the hCAR LBD-containing polypeptide that is less than that bound to the hCAR LBD-containing polypeptide in the absence of ligand indicates that the ligand inhibits binding of clotrimazole to the hCAR LBD-containing polypeptide and is non-specific in its binding to the selected nuclear receptor.
A method of screening a selected compound for ability to bind a human Constitutive Androstane Receptor (hCAR) ligand binding domain (LBD) comprising
- i) contacting the selected compound and clotrimazole with the hCAR LBD-containing polypeptide under conditions such that clotrimazole can bind the hCAR LBD-containing polypeptide in the absence of the selected compound, and
- ii) determining the amount of clotrimazole bound to the hCAR LBD-containing polypeptide,
- wherein an amount of clotrimazole bound to the hCAR LBD-containing polypeptide that is less than that bound to the hCAR LBD-containing polypeptide in the absence of the selected compound indicates that the selected compound binds the hCAR LBD.
Dependencies are inferred from the claim text.