Assays For Detecting Inhibitors Of Binding Between COX-2 And PDZ Proteins

Abstract

The invention provides an assay for determining whether a test agent is a COX modulator. In general terms, the assay includes: determining whether a test agent modulates binding of a PDZ-containing polypeptide to a COX PL-containing polypeptide. The PDZ-containing polypeptide may contain the PDZ domain of PDZ domain of MAGI1, TIP-1, MAST2, PSD95, or SHANK. The assays may be done in a cell-free environment or in a cellular environment, particularly using a neuronal cell. The invention finds use in a variety of therapeutic applications, including for identifying agents for use in treating cancer, pain, inflammation and neuronal conditions caused by acute insult, e.g., stroke.

1 . An assay for detecting a COX modulator, comprising:
 determining whether a test agent modulates binding of a PDZ-containing polypeptide to a COX PDZ ligand-containing polypeptide.   
     
     
         2 . The assay of  claim 1 , wherein said COX PDZ ligand-containing polypeptide is a COX-2 PDZ ligand-containing polypeptide. 
     
     
         3 . The assay of  claim 1 , wherein said PDZ-containing polypeptide contains a PDZ domain of MAGI1, TIP-1, MAST2, PSD95, or SHANK. 
     
     
         4 . The assay of  claim 3 , wherein said SHANK PDZ-containing polypeptide comprises a PDZ domain of SHANK1, SHANK2 or SHANK-3. 
     
     
         5 - 6 . (canceled) 
     
     
         7 . The assay of  claim 1 , wherein said assay is a cell-free assay. 
     
     
         8 . The assay of  claim 1 , wherein said assay is a cellular assay. 
     
     
         9 - 10 . (canceled) 
     
     
         11 . The assay of  claim 8 , wherein said assay is performed using neuronal cells that contain said PDZ domain-containing polypeptide and said COX PDZ ligand-containing polypeptide. 
     
     
         12 . The assay of  claim 1 , wherein said assay further comprises testing said agent for COX-2 cycloxygenase inhibitory activity. 
     
     
         13 . The assay of  claim 1 , wherein said test agent is an inhibitor of a cycloxygenase activity of COX-2. 
     
     
         14 . The assay of  claim 1 , wherein said test agent is PDZ domain analog. 
     
     
         15 . The assay of  claim 1 , further comprises testing said compound in a neuronal cell. 
     
     
         16 . The assay of  claim 15 , further comprising subjecting said neuronal cell to insult. 
     
     
         17 . The assay of  claim 15 , wherein said insult is hypoxia or ischemia. 
     
     
         18 . A method of reducing binding between COX-2 and a PDZ-containing polypeptide in a cell, comprising:
 administering to said cell a PDZ domain analog or a compound which competes with the binding of COX-2 to said PDZ-containing polypeptide; and   maintaining said cell under conditions suitable for said PDZ domain analog or compound to reduce said binding.   
     
     
         19 - 23 . (canceled) 
     
     
         24 . The method of  claim 18 , wherein said cell is an insulted neuronal cell. 
     
     
         25 . The method of  claim 24 , wherein said neuronal cell is a hypoxic or ischemic neuronal cell. 
     
     
         26 . The method of  claim 24 , wherein said method results in reduced NMDA receptor activation. 
     
     
         27 . The method of  claim 18 , wherein said administration comprises administering a compound selected from the group consisting of: sulindac sulphide, fenoprofen, derivatives thereof, analogs thereof, and combinations thereof. 
     
     
         28 . The method of  claim 18 , wherein said reduction in binding further results in anti-tumor and/or anti-cellular proliferate properties when administered in vivo. 
     
     
         29 . The method of  claim 28 , wherein said PDZ domain analog or said compound which competes with the binding of COX-2 to said PDZ-containing polypeptide is administered to a subject suffer from cancer, and said anti-tumor and/or anti-cellular proliferate properties results in treatment of said cancer.