Compositions And Methods For Identifying Modulars Of TRPV2

Abstract

It has now been discovered that certain cannabinoids specifically activate TRPV2 channel activity. Based on the discovery, novel compositions and methods for screening, identifying and characterizing compounds that increase or decrease the biological activity of a TRPV2.

1 - 18 . (canceled) 
     
     
         19 . A method for identifying a compound that increases the biological activity of TRPV2, comprising the steps of
 a. obtaining atomic coordinates defining a three-dimensional structure of a complex comprising a TRPV2 interacting with a cannabinoid that is capable of activating the TRPV2;   b. elucidating a structural relationship between the TRPV2 and the interacting cannabinoid;   c. designing a structural analog of the cannabinoid based on the structural relationship;   d. synthesizing the structural analog;   e. determining the extent to which the structural analog alters the biological activity of the TRPV2, thereby identifying the compound that increases the biological activity of TRPV2.   
     
     
         20 . The method of  claim 19 , wherein the biological activity of the TRPV2 is determined as calcium-influx into a cell expressing the TRPV2. 
     
     
         21 . The method of  claim 19 , wherein the biological activity of the TRPV2 is measured by a method of patch clamp. 
     
     
         22 . The method of  claim 19 , wherein the biological activity of the TRPV2 is measured by a CA mobilization assay. 
     
     
         23 . The method of  claim 19 , wherein the biological activity of the TRPV2 is determined as its binding affinity to the cannabinoid that is capable of activating the TRPV2 activity. 
     
     
         24 . A method of increasing the biological activity of a TRPV2, comprising the step of contacting the TRPV2 with a cannabinoid that is capable of activating the TRPV2 activity. 
     
     
         25 . The method of  claim 24 , wherein the TRPV2 is associated with an isolated membrane. 
     
     
         26 . The method of  claim 24 , wherein the TRPV2 is present in a cell. 
     
     
         27 . The method of  claim 26 , wherein the cell is a neuron. 
     
     
         28 . The method of  claim 24 , wherein the cannabinoid is selected from the group consisting of Δ 9 -tetrahydrocannabinol, 11-hydroxy-Δ 9 -tetrahydrocannabinol, cannabinol, cannabidiol, O-1821, nabilone, CP55940, 2-AG, and HU210, HU211, HU308, and HU331. 
     
     
         29 . A method for stimulating noxious thermo-sensation in a subject, comprising administering to the subject a pharmaceutical composition comprising an effective amount of cannabinoid that is capable of activating the TRPV2 activity, thereby stimulating the noxious thermo-sensation in the subject. 
     
     
         30 . The method of  claim 29 , wherein the subject is a human. 
     
     
         31 . The method of  claim 29 , wherein the cannabinoid is selected from the group consisting of Δ 9 -tetrahydrocannabinol, 11-hydroxy-Δ 9 -tetrahydrocannabinol, cannabinol, cannabidiol, O-1821, nabilone, CP55940, 2-AG, and HU210, HU211, HU308, and HU331. 
     
     
         32 . An isolated polypeptide consisting essentially of an amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17. 
     
     
         33 . An isolated nucleic acid molecule that encodes a polypeptide consisting essentially of an amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17. 
     
     
         34 . The isolated nucleic acid molecule of  claim 33  consisting essentially of a nucleotide sequence of SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO:26, SEQ ID NO:27, or SEQ ID NO:28. 
     
     
         35 . An expression vector comprising nucleotide sequence that encodes a polypeptide consisting essentially of an amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17. 
     
     
         36 . A recombinant cell comprising an expression vector of  claim 35 . 
     
     
         37 . A method of producing a polypeptide consisting essentially of an amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17, comprising the step of growing a cell of  claim 36  under a condition whereby the polypeptide is produced by the cell.