Assays for fprl-1 ligands
AssigneeAstraZeneca A B
InventorsDuncan Henderson, Sylvia Salter, Aram Elagoz, Eric Grazzni, Paola Lembo
Life of this patent
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Abstract
Assays for screening for compounds that act as agonists, antagonists, or inverse agonists of CKβ8-1 at the FPRL-1 receptor.
Claims33
A method for identifying compounds that modulate binding of CKβ8-1 to FPRL-1 receptor, said method comprising:
- providing cells expressing FPRL-1 receptor or functional fragment or variant thereof;
- contacting said cells with CKβ8-1 or a functional fragment or variant thereof, in the presence or absence of a compound;
- and
- measuring a signal indicative of receptor activation;
- where an alteration to said signal in the presence of a compound identifies said compound as a compound that modulates binding of CKβ8-1 to FPRL-1 receptor.
A method for identifying compounds that modulate the binding of CKβ8-1 to the FPRL-1 receptor, said method comprising:
- providing the FPRL-1 receptor or functional fragment or variant thereof;
- contacting the FPRL-1 receptor or functional fragment or variant thereof, with CKβ8-1 or functional fragment or variant thereof in the presence or absence of a compound;
- and
- measuring the amount of CKβ8-1 or functional fragment or variant thereof that forms a complex with the FPRL-1 receptor or functional fragment or variant thereof;
- where an alteration to the amount of said complex formed in the presence of said compound identifies said compound as a compound that modulates binding of CKβ8-1 to the FPRL-1 receptor.
A method of screening for a FPRL-1 receptor agonist or antagonist comprising measuring a cell stimulating activity through a FPRL-1 receptor determined from the following steps a) and b);
- a) contacting a compound with a cell expressing a FPRL-1 receptor or functional fragment or variant thereof (test screen), and comparing the results to a control screen wherein the cell does not express the FPRL-1 receptor or functional fragment or variant thereof, wherein said compound having cell stimulating activity in the test screen but not the control screen indicates that the test compound is a FPRL-1 receptor agonist,
- b) contacting CKβ8-1 or functional fragment or variant thereof and a test compound with a cell expressing a FPRL-1 receptor or functional fragment or variant thereof (test screen), and comparing the results to a control screen wherein the cell does not express the FPRL-1 receptor or functional fragment or variant thereof, where a decrease in cell stimulating activity by CKβ8-1 or functional fragment or variant thereof in the test screen but not the control screen indicates that the test compound is a FPRL-1 receptor antagonist.
A method of screening for compounds that modulate binding of CKβ8-1 to FPRL-1 receptor, comprising comparing the amount of CKβ8-1 or functional fragment or variant thereof bound to FPRL-1 receptor or functional fragment or variant thereof in steps a) and b):
- a) contacting CKβ8-1 or functional fragment or variant thereof with the FPRL-1 receptor or functional fragment or variant thereof;
- b) contacting CKβ8-1 or functional fragment or variant thereof and a test compound with the FPRL-1 receptor or functional fragment or variant thereof;
- where an alteration in the amount of CKβ8-1 or functional fragment or variant thereof bound to FPRL-1 receptor or functional fragment or variant thereof in step b) indicates that the test compound modulates binding of CKβ8-1 to the FPRL-1 receptor.
A method of screening for compounds that inhibit binding of CKβ8-1 to FPRL-1 receptor, comprising comparing the amount of CKβ8-1 or functional fragment or variant thereof bound to FPRL-1 receptor or functional fragment or variant thereof in steps a) and b):
- a) contacting CKβ8-1 or functional fragment or variant thereof with the FHRL-1 receptor or functional fragment or variant thereof;
- b) contacting CKβ8-1 or functional fragment or variant thereof and a test compound with the FPRL-1 receptor or functional fragment or variant thereof;
- where a decrease in CKβ8-1 or functional fragment or variant thereof binding in step b) indicates that the test compound inhibits binding of CKβ8-1 to the FPRL-1 receptor.
A method of identifying a compound that modulates binding of CKβ8-1 to FPRL-1 receptor, comprising contacting FPRL-1 receptor or functional fragment or variant thereof with CKβ8-1 or functional fragment or variant thereof in the presence or absence of a test compound, and comparing the amount of binding between CKβ8-1 or functional fragment or variant thereof and the FPRL-1 receptor or functional fragment or variant thereof in the presence or absence of the test compound, where an alteration in the amount of binding between CKβ8-1 or functional fragment or variant thereof and the FPRL-1 receptor or functional fragment or variant thereof in the presence of the test compound indicates that the test compound modulates binding between CKβ8-1 and the FPR-1 receptor.
A method of identifying a compound that binds FPRL-1 receptor, comprising incubating a cell expressing FPRL-1 receptor or functional fragment or variant thereof with CKβ8-1 or functional fragment or variant thereof in the presence or absence of a compound, and detecting displacement of CKβ8-1 or functional fragment or variant thereof binding to the FPRL-1 receptor or functional fragment or variant thereof in the presence of the compound, where displacement of said binding is indicative of a compound that binds the FPRL-1 receptor.
A method of determining if a test compound is an agonist, antagonist or inverse agonist of CKβ8-1 comprising
- a) incubating a cell expressing FPRL-1 or functional fragment or variant thereof with the test compound;
- b) measuring a signal indicative of receptor activation;
- and
- c) comparing the measurement in b) with a second measurement of a signal indicative of receptor activation obtained from incubations performed in the absence of the test compound, where the test compound is an agonist of CKβ8-1 if the signal indicative of receptor activation is higher in the presence of the test compound than in its absence, and wherein the test compound is an antagonist of CKβ8-1 if the signal indicative of receptor activation is lower in the presence of the test compound than in its absence.
Dependencies are inferred from the claim text.