Methods for detecting receptor modulator activity

• US 7,655,396 B1
• Filed: 09/14/2007
• Issued: 02/02/2010
• Est. Priority Date: 09/29/2006
• Status: Expired due to Fees

• Current Assignee: Allergan, Inc. (Canada) (Abbvie Incorporated)
• Original Assignee: Allergan, Inc. (Canada) (Abbvie Incorporated)
• Inventors: Kedzie, Karen M., Rao, Sandhya, Donello, John E., Gil, Daniel W.
• Primary Examiner(s): Li; Ruixiang
• Application Number:
  US11/855,858
• Time in Patent Office: 872 Days
• Field of Search: None
• US Class Current: 435/4
• CPC Class Codes: G01N 2333/726 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
• CPC Class Codes: G01N 2500/10 involving cells
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds

Abstract

Disclosed herein are methods of screening compounds that modulate G-protein coupled receptors. The assays are performed at temperatures of between about 25° C. and 40° C.

Claims

1. A high-throughput method for identifying partial agonists and partial antagonists of a G-protein coupled receptor, the method comprising the steps of

a) contacting, at a temperature of about 25°

C., a test compound with a cell expressing the receptor;

b) detecting a change in the level of activity of the receptor;

c) contacting, at a temperature of about 35°

C., a test compound with a cell expressing the receptor;

d) detecting a change in the level of activity of the receptor

e) comparing the result obtained in step b) with the result obtained in step d) wherein a partial agonist or antagonist of the G-protein coupled receptor is identified if a compound shows no activity at a temperature of about 25°

C. but does show activity at a temperature of about 37°

C.

2. The method of claim 1 in which both of steps b) and d) comprises detecting a change in intracellular ion flux in said cell.

3. The method of claim 1 in which both of steps b) and d) comprises detecting a change in the level of intracellular second messengers in said cell.

4. The method of claim 1 wherein the G-protein coupled receptor is selected from the group consisting of an alpha adrenergic receptor, a beta adrenergic receptor, a cannabinoid receptor, a chemokine receptor, a dopamine receptor, an endothelial differentiation sphingolipid receptor, a 5-hydroxytryptamine G-protein coupled receptor, a y-aminobutyric acid G-protein coupled receptor, a histamine receptor, a metabotropic glutamate receptor, an opioid receptor, a prostaglandin receptor, a somatostatin receptor, and a tachykinin receptor.