Method for screening compounds for alpha1B adrenergic receptor antagonist and analgesic activity

Abstract

A method for identifying compounds that can be useful for producing analgesia is disclosed. In particular, the method involves providing a preparation of a cell, that expresses an alpha 1B adrenergic receptor, combining a test compound with the cell preparation, measuring the effect of the test compound on the alpha 1B receptor activity, and evaluating the compound thus identified for analgesic activity.

What is claimed is:  
     
         1 . A method for identifying a compound that modulates alpha 1B  adrenergic receptor activity, comprising: 
 (a) measuring the activity of a test compound in a first binding assay, comprising 
 (i) providing a preparation of a cell that expresses an alpha 1B  adrenergic receptor,  
 (ii) combining the test compound with the cell preparation,  
 (iii) measuring binding of the test compound to the cell preparation or the receptor;  
   (b) measuring activity of the test compound in at least one pain model; and    (c) measuring the activity of the test compound in a second binding assay, comprising 
 (i) providing a preparation of a cell that expresses an alpha 1A  adrenergic receptor, a preparation of a cell that expresses an alpha 1D  adrenergic receptor, or a preparation of a cell that expresses an alpha 1A  adrenergic receptor and a preparation of a cell that expresses an alpha 1D  adrenergic receptor,  
 (ii) combining the test compound with the cell preparation, and  
 (iii) measuring the binding of the test compound to the cell or the receptor;  
   wherein (a), (b) and (c) are done concurrently or consecutively in any order.    
     
     
         2 . The method of  claim 1 , wherein the preparation of the cell that expresses the alpha 1B  adrenergic receptor is a membrane preparation or an intact cell preparation.  
     
     
         3 . The method of  claim 1 , wherein the pain model is an animal pain model.  
     
     
         4 . The method of  claim 3 , wherein the animal pain model is a tail flick model, a rat tail immersion model, a carrageenan-induced paw hyperalgesia model, a formalin behavior response model, a von Frey filament test, a radiant heat model, or a cold allodynia model.  
     
     
         5 . The method of  claim 1 , further comprising measuring the effect of the test compound on general locomotor activity.  
     
     
         6 . The method of  claim 1 , further comprising measuring the effect of the test compound on sedation.  
     
     
         7 . The method of  claim 1 , further comprising measuring a functional alpha 1 -adrenergic receptor activity of the test compound.  
     
     
         8 . The method of  claim 7 , wherein the functional alpha 1 -receptor activity of the test compound is measured in an in vitro test.  
     
     
         9 . The method of  claim 7  wherein the functional alpha 1 -receptor activity of the test compound is measured in an in vivo test.  
     
     
         10 . A method of treating a subject to produce analgesia comprising administering to a subject in need of analgesia a therapeutically effective amount of a compound identified by the method of  claim 1 , or a pharmaceutically acceptable salt or hydrate thereof.  
     
     
         11 . A method of treating a subject to produce analgesia, comprising administering to a subject in need of analgesia a therapeutically effective amount of a composition comprising a compound identified by the method of  claim 1 , or a pharmaceutically acceptable salt or hydrate thereof and a pharmaceutically acceptable carrier.  
     
     
         12 . A method of treating a subject to produce analgesia, comprising administering to a subject in need of analgesia a therapeutically effective amount of a compound identified by the method of  claim 7 , or a pharmaceutically acceptable salt or hydrate thereof.  
     
     
         13 . A method of treating a subject to produce analgesia, comprising administering to a subject in need of analgesia a therapeutically effective amount of a composition comprising a compound identified by the method of  claim 7 , or a pharmaceutically acceptable salt or hydrate thereof and a pharmaceutically acceptable carrier.  
     
     
         14 . The method of  claim 11 , wherein the composition further comprises one or more compounds having at least one of anti-inflammatory activity, analgesic activity, or anticonvulsant activity.  
     
     
         15 . The method of  claim 13 , wherein the composition further comprises one or more compounds having at least one of anti-inflammatory activity, analgesic activity, or anticonvulsant activity.  
     
     
         16 . A pharmaceutical composition, comprising a therapeutically effective amount of a compound identified by the method of  claim 1 , or a pharmaceutically acceptable salt or hydrate thereof, and one or more pharmaceutically acceptable additives, diluents, or carriers.  
     
     
         17 . A pharmaceutical composition, comprising a therapeutically effective amount of a compound identified by the method of  claim 7 , or a pharmaceutically acceptable salt or hydrate thereof, and one or more pharmaceutically acceptable additives, diluents, or carriers.  
     
     
         18 . A method of treating a subject suffering from a CNS disorder comprising administering to the, subject a therapeutically effective amount of a compound identified by the method of  claim 1  or a pharmaceutically acceptable salt or hydrate thereof.  
     
     
         19 . A method of treating a subject suffering from a CNS disorder comprising administering to the subject a therapeutically effective amount of a composition comprising a compound identified by the method of  claim 1  or a pharmaceutically acceptable salt or hydrate thereof and a pharmaceutically acceptable carrier.