Methods and reagents for discovering and using mammalian melanocortin receptor agonists and antagonists to modulate feeding behavior in animals

Abstract

The present invention provides recombinant expression constructs comprising nucleic acid encoding mammalian melanocortin receptors, and mammalian cells into which said recombinant expression constructs have been introduced that express functional mammalian melanocortin receptors. The invention provides a panel of such transformed mammalian cells expressing melanocortin receptors for screening compounds for receptor agonist and antagonist activity. The invention also provides methods for using such panels of melanocortin receptor-expressing mammalian cells to specifically detect and identify agonists and antagonists for each melanocortin receptor, as well as patterns of agonist and antagonist activity of said compounds for the class of melanocortin receptors. Such screening methods provide a means for identifying compounds with patterns of melanocortin agonist and antagonist activity which is associated with the capacity to influence or modify metabolism and behavior, particularly feeding behavior.

We claim:  
     
         1 . A compound having the general formula:  
       A-B-C-D-E-F-G-amide  
       wherein 
 A is Leu, Ile, Met, or isosteres thereof;  
 B is Asp, Glu, or substituted analogues thereof;  
 C is His or isosteres thereof;  
 D is  d -Phe,  d -Tyr or isosteres thereof;  
 E is Arg, Lys, homoArg, homoLys, or isosteres thereof;  
 F is Trp or isosteres thereof;  
 G is Lys, homoLys or isosteres thereof;  
 and wherein the peptide is cyclized by the formation of an amide bond between the side chain carboxyl group of the Asp or Glu residue at position B in the peptide, and the side chain amino group of the Lys or homoLys residue at position G, and wherein the compound is a mammmalian melanocortin MC-3 or MC-4 receptor agonist.  
 
     
     
         2 . A compound having the general formula:  
       A-B-C-D-E-F-G-amide  
       wherein 
 A is Leu, Ile, Met, or isosteres thereof;  
 B is Asp, Glu, or isosteres thereof;  
 C is His, or isosteres thereof;  
 D is  d -Nal, or isosteres thereof;  
 E is Arg, Lys, homoArg, homoLys, or isosteres thereof;  
 F is Trp, or isosteres thereof;  
 G is Lys, homoLys, or isosteres thereof;  
 and wherein the peptide is cyclized by the formation of an amide bond between the side chain carboxyl group of the Asp or Glu residue at position B in the peptide, and the side chain amino group of the Lys or homoLys residue at position G, and wherein the compound is a mammalian melanocortin MC-3 or MC-4 receptor antagonist.