Methods for identification of modulators of OSGPR116 activity, and their use in the treatment of disease

Abstract

This invention relates to the identification of fatty acid or lipid amides that decrease food intake in mammals, including fatty acid ethanolamides, as ligands for the G-protein coupled receptor OSGPR116, and describes the first demonstration of a specific G-protein coupled receptor that is activated by fatty acid ethanolamides that inhibit feeding. The invention is directed to new methods for screening candidate drugs for their ability to modulate the activity of OSGPR116, and new pharmaceutical agents identified by these methods. It is also directed to the use of such agents in the manufacture of medicaments for the treatment of OSGPR116 mediated diseases, and methods of treating diseases such as obesity and diabetes by administering to an individual a therapeutic amount of a modulator of OSGPR116 identified by these methods.

1 - 64 . (canceled)  
     
     
         65 . A method involving competitive binding for identifying a chemical compound which specifically binds to a mammalian OSGPR116 receptor which comprises separately contacting cells expressing on their cell surface the mammalian OSGPR116 receptor, wherein such cells do not normally express the mammalian OSGPR116 receptor, with both the chemical compound and a lipid amide or fatty acid amide ligand, known to bind to the receptor, and with only the ligand, under conditions suitable for binding of such a ligand to the receptor, and detecting specific binding of the chemical compound to the mammalian OSGPR116 receptor, a decrease in the binding of the ligand to the mammalian OSGPR116 receptor in the presence of the chemical compound being tested indicating that such chemical compound binds to the mammalian OSGPR116 receptor.  
     
     
         66 . The method of  claim 65 , wherein the ligand for OSGPR116 is a fatty acid amide selected from oleoyl ethanolamide, palmitoyl ethanolamide, stearoyl ethanolamide, linoleoyl ethanolamide, linolenoyl ethanolamide and oleamide.  
     
     
         67 . The method of  claim 65 , wherein the ligand for OSGPR116 is oleoyl ethanolamide.  
     
     
         68 . The method of  claim 65 , wherein the mammalian OSGPR116 is a human OSGPR116 receptor.  
     
     
         69 . The method of  claim 65 , wherein the cell is an insect cell.  
     
     
         70 . The method of  claim 65 , wherein the cell is a mammalian cell.  
     
     
         71 . The method of  claim 65 , wherein the cell is a human cell.  
     
     
         72 . The method of  claim 65 , wherein the cell is a yeast cell.  
     
     
         73 . The method of  claim 70 , wherein the cell is a COS-7 cell, 293 human embryonic kidney cell, a CHO cell, a NIH-3T3 cell, a mouse Y1 cell, an adipose cell, or a pancreatic cell.  
     
     
         74 . A method involving competitive binding for identifying a chemical compound which specifically binds to a mammalian OSGPR116 receptor which comprises separately contacting a membrane preparation from cells expressing on their cell surface the mammalian OSGPR116 receptor, wherein such cells do not normally express the mammalian OSGPR116 receptor, with both the chemical compound and a lipid amide or fatty acid amide ligand, known to bind to the receptor, and with only the ligand, under conditions suitable for binding of such a ligand to the receptor, and detecting specific binding of the chemical compound to the mammalian OSGPR116 receptor, a decrease in the binding of the ligand to the mammalian OSGPR116 receptor in the presence of the chemical compound being tested indicating that such chemical compound binds to the mammalian OSGPR116 receptor.  
     
     
         75 . The method of  claim 74 , wherein the ligand for OSGPR116 is a fatty acid amide selected from oleoyl ethanolamide, palmitoyl ethanolamide, stearoyl ethanolamide, linoleoyl ethanolamide, linolenoyl ethanolamide and oleamide.  
     
     
         76 . The method of  claim 74 , wherein the ligand for OSGPR116 is oleoyl ethanolamide.  
     
     
         77 . The method of  claim 74 , wherein the mammalian OSGPR116 is a human OSGPR116 receptor.  
     
     
         78 . The method of  claim 74 , wherein the cell is an insect cell.  
     
     
         79 . The method of  claim 74 , wherein the cell is a mammalian cell.  
     
     
         80 . The method of  claim 74 , wherein the cell is a human cell.  
     
     
         81 . The method of  claim 74 , wherein the cell is a yeast cell.  
     
     
         82 . The method of  claim 79 , wherein the cell is a COS-7 cell, 293 human embryonic kidney cell, a CHO cell, a NIH-3T3 cell, a mouse Y1 cell, an adipose cell, or a pancreatic cell.  
     
     
         83 . A method of screening a plurality of chemical compounds not known to bind to a mammalian OSGPR116 receptor to identify a compound which specifically binds to the mammalian OSGPR116 receptor, which comprises 
 (a) contacting cells transfected with and expressing DNA encoding the mammalian OSGPR116 receptor with a lipid amide or fatty acid amide ligand known to bind specifically to the mammalian OSGPR116 receptor;    (b) contacting the cells of step (a) with the plurality of compounds not known to bind specifically to the mammalian OSGPR116 receptor, under conditions permitting binding of compounds known to bind to the mammalian OSGPR116 receptor;    (c) determining whether the binding of the lipid amide or fatty acid amide ligand known to bind to the mammalian OSGPR116 receptor is reduced in the presence of the plurality of compounds, relative to the binding of the lipid amide or fatty acid amide ligand in the absence of the plurality of compounds; and if so    (d) separately determining the binding to the mammalian OSGPR116 receptor of each compound included in the plurality of compounds, so as to thereby identify any compound included therein which specifically binds to the mammalian OSGPR116 receptor;    
     
     
         84 . The method of  claim 83  wherein the mammalian OSGPR116 receptor is a human OSGPR116 receptor.  
     
     
         85 . The method of  claim 83 , wherein the cell is a mammalian cell.  
     
     
         86 . The method of  claim 83 , wherein the cell is human cell.  
     
     
         87 . The method of  claim 83 , wherein the cell is yeast cell.  
     
     
         88 . The method of  claim 83 , wherein the cell is a COS-7 cell, a 293 human embryonic kidney cell, a CHO cell, a mouse Y1 cell, an adipose cell, a pancreatic cell, or an NIH-3T3 cell.  
     
     
         89 . A method for identification of an agent that inhibits feeding, which method comprises contacting a subject with a test agent identified by the method of any one of claims  65 ,  74 , and  83 , and monitoring feeding, thereby determining whether the test agent is an inhibitor of feeding.  
     
     
         90 . A method for identification of an agent that causes a reduction in obesity in a subject, which method comprises contacting a subject with a test agent identified by the method of any one of claims  65 ,  74 , and  83 , and monitoring obesity, thereby determining whether the test agent is an inhibitor of obesity.  
     
     
         91 . A method of preparing a composition comprising a chemical compound which specifically binds to a mammalian OSGPR116 receptor, which comprises separately contacting cells expressing on their cell surface the mammalian OSGPR116 receptor, wherein such cells do not normally express the mammalian OSGPR116 receptor, with both a test chemical compound and a lipid amide or fatty acid amide ligand, known to bind to the receptor, and with only the ligand, under conditions suitable for binding of such a ligand to the receptor, and detecting specific binding of the test chemical compound to the mammalian OSGPR116 receptor, a decrease in the binding of the ligand to the mammalian OSGPR116 receptor in the presence of the test chemical compound indicating that said test chemical compound binds specifically to the mammalian OSGPR116 receptor, and admixing the test chemical so identified, or a functional analog or homolog of said test chemical, with a carrier, thereby preparing said composition.  
     
     
         92 . The method of claim  95 , wherein the mammalian OSGPR116 receptor is a human OSGPR116 receptor.  
     
     
         93 . A method of preparing a composition comprising a chemical compound which specifically binds to a mammalian OSGPR116 receptor, which comprises separately contacting a membrane preparation from cells expressing on their cell surface the mammalian OSGPR116 receptor, wherein such cells do not normally express the mammalian OSGPR116 receptor, with both a test chemical compound and a lipid amide or fatty acid amide ligand, known to bind to the receptor, and with only the ligand, under conditions suitable for binding of such a ligand to the receptor, and detecting specific binding of the test chemical compound to the mammalian OSGPR116 receptor, a decrease in the binding of the ligand to the mammalian OSGPR116 receptor in the presence of the test chemical compound indicating that said test chemical compound binds specifically to the mammalian OSGPR116 receptor, and admixing the test chemical so identified, or a functional analog or homolog of said test chemical, with a carrier, thereby preparing said composition.  
     
     
         94 . The method of claim  97 , wherein the mammalian OSGPR116 receptor is a human OSGPR116 receptor.