Assay for and uses of peptide hormone receptor agonists

Abstract

The invention features a method for determining whether a candidate compound is a non-peptide agonist of a peptide hormone receptor. In this method, a candidate compound is exposed to a form of the peptide hormone receptor, or to a protein that interacts with a peptide hormone receptor, which has an enhanced ability to amplify the intrinsic activity of a non-peptide agonist. The second messenger signaling activity of the enhanced receptor is measured in the presence of the candidate compound, and compared to the second messenger signaling activity of the wildtype receptor measured in the absence of the candidate compound. A change in second messenger signaling activity indicates that the candidate compound is an agonist. An increase in second messenger signaling activity indicates that the compound is either a full or partial positive agonist; a decrease in second messenger signaling activity indicates that the compound is an inverse (also termed a ‘negative’) agonist. The invention further embraces a method of using a peptide hormone receptor agonist for the treatment or prevention of a physiological disease, as well as particular enhanced receptors and the nucleic acid sequences which code for them.

What is claimed is:  
     
         1 . A method for treating or preventing a physiological disorder involving a CCK or gastrin peptide hormone receptor, said method comprising administering to a mammal an agonist-effective amount of a compound of formula (I):  
       
         
           
           
               
               
           
         
         wherein: 
 R 1  represents H, C 1-6  alkyl optionally substituted by one or more halo groups, C 3-7  cycloalkyl, cyclopropylmethyl, (CH 2 ) r imidazolyl, (CH 2 ) r triazolyl, (CH 2 ) r tetrazolyl, where r is 1, 2 or 3, CH 2 CO 2 R 11 , where R 11  is C 1-4 alkyl, or CH 2 CONR 6 R 7 , where R 6  and R 7  each independently represent H or C 1-4 alkyl, or R 6  and R 7  together form a chain CH 2(p)  where p is 4 or 5;  
 R 2  represents NHR 12  or (CH 2 ) s R 13 , where s is 0, 1, 2, or 3;  
 R 3  represents C 1-6 alkyl, halo, or NR 6 R 7 , where R 6  and R 7  are as previously defined;  
 R 4  and R 5  each independently represent H, C 1-12 alkyl optionally substituted by NR 9 R 9 ′, where R 9  and R 9 , are each independently H or C 1-4 alkyl, an azacyclic or azabicyclic group, C 4-9 cycloalkyl optionally substituted by one or more C 1-4 alkyl groups, C 4-9 cycloalkyl, C 1-4 alkyl optionally substituted in the cycloalkyl ring by one or more C 1-4 alkyl groups, optionally substituted aryl, optionally substituted arylC 1-6 alkyl or azacyclic or azabicyclic groups, or R 4  and R 5  together form the residue of an optionally substituted azacyclic or azabicyclic ring system;  
 x is 0, 1, 2, or 3;  
 R 12  represents a phenyl or pyridyl group optionally substituted by one or more substituents selected form C 1-6 alkyl, halo, hydroxy, C 1-4 alkoxy, (CH 2 ) q -tetrazolyl optionally substituted in the tetrazole ring by C 1-4 alkyl, (CH 2 ) q -imidazolyl, (CH 2 ) q -triazolyl, where q is 0, 1, 2, or 3, 5-hydroxy-4-pyrone, NR 6 R 7 , NR 9 COR 11 , NR 9 CONR 9 ′R 11 , where R 6 , R 7 , R 9 , R 9 ′, and R 11  are each as previously defined, SO(C 1-6 alkyl), SO 2 (C 1-6 alkyl), trifluoromethyl, CONHSO 2 R 8  or SO 2 NHCOR 8 , where R 8  is C 1-6 alkyl, optionally substituted aryl, 2,2-difluorocyclopropane or trifluoromethyl, SO 2 NHR 10 , where R 10  is a nitrogen containing heterocycle, B(OH) 2 , (CH 2 ) q CO 2 H, where q is as previously defined; or  
 R 12  represents a group:  
                     
 where X 1  represents CH or N; W represents CH 2  or NR 9 , where R 9  is as previously defined, and W 1  represents CH 2 , or W and W 1  each represent O; or  
 R 12  represents phenyl substituted by a group:  
                     
 wherein X 2  is O, S, or NR 9 , where R 9  is as previously defined; z is a bond, O, or S; m is 1, 2 or 3; n is 1, 2, or 3; and y is 0, 1, 2, or 3;  
 
         R 13  represents a group:  
         
           
             
             
                 
                 
             
           
         
         where R 14  represents H or C 1-6 alkyl; R 15  represents H, C 1-6 alkyl, halo or NR 6 R 7 , where R 6  and R 7  are as previously defined; and the dotted line represents an optional covalent bond; and pharmaceutically acceptable salts or prodrugs thereof,  
         with the provision that, when NR 4 R 5  represents an unsubstituted azacyclic ring system, R 2  does not represent NHR 12  where R 12  is optionally substituted phenyl or:  
         
           
             
             
                 
                 
             
           
         
         where W and W 1  are as previously defined.  
       
     
     
         2 . The method of  claim 1 , wherein said compound is 3(R,S)-Amino-1,3-dihydro-5-((1S,4S)-5-methyl-2,5-diazabicyclo[2,2,1]heptan-2-vl)-2H-1-propyl-1,4-benzodiazepin-2-one.  
     
     
         3 . The method of  claim 1 , wherein said compound is (−)-N-[5-(3-azabicyclo[3,2,2]nonan-3-yl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[3-methylphenyl]-urea.  
     
     
         4 . The method of  claim 1 , wherein said compound is (+)-N-[5-(3-azabicyclo[3,2,2]nonan-3-yl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[3-methylphenyl]-urea.  
     
     
         5 . The method of  claim 1 , wherein said peptide hormone receptor is the CCK-B/gastrin receptor.  
     
     
         6 . The method of  claim 1 , wherein said peptide hormone receptor is the CCK-A receptor.  
     
     
         7 . The method of  claim 1 , wherein said physiological disorder is a neoplasm.  
     
     
         8 . The method of  claim 1 , wherein said neoplasm is a primary tumor.  
     
     
         9 . A compound of formula (I):  
       
         
           
           
               
               
           
         
         wherein: 
 R 1  represents H, C 1-6  alkyl optionally substituted by one or more halo groups, C 3-7  cycloalkyl, cyclopropylmethyl, (CH 2 ) r imidazolyl, (CH 2 ) r triazolyl, (CH 2 ) r tetrazolyl, where r is 1, 2 or 3, CH 2 CO 2 R 11 , where R 11  is C 1-4 alkyl, or CH 2 CONR 6 R 7 , where R 6  and R 7  each independently represent H or C 1-4 alkyl, or R 6  and R 7  together form a chain CH 2(p)  where p is 4 or 5;  
 R 2  represents NHR 12  or (CH 2 ) s R 13 , where s is 0, 1, 2, or 3;  
 R 3  represents C 1-6 alkyl, halo, or NR 6 R 7 , where R 6  and R 7  are as previously defined;  
 R 4  and R 5  each independently represent H, C 1-12 alkyl optionally substituted by NR 9 R 9 ′, where R 9  and R 9 , are each independently H or C 1-4 alkyl, an azacyclic or azabicyclic group, C 4-9 cycloalkyl optionally substituted by one or more C 1-4 alkyl groups, C 4-9 cycloalkyl, C 1-4 alkyl optionally substituted in the cycloalkyl ring by one or more C 1-4 alkyl groups, optionally substituted aryl, optionally substituted arylC 1-6 alkyl or azacyclic or azabicyclic groups, or R 4  and R 5  together form the residue of an optionally substituted azacyclic or azabicyclic ring system;  
 x is 0, 1, 2, or 3;  
 R 12  represents a phenyl or pyridyl group optionally substituted by one or more substituents selected form C 1-6 alkyl, halo, hydroxy, C 1-4 alkoxy, (CH 2 ) q -tetrazolyl optionally substituted in the tetrazole ring by C 1-4 alkyl, (CH 2 ) q -imidazolyl, (CH 2 ) q -triazolyl, where q is 0, 1, 2, or 3, 5-hydroxy-4-pyrone, NR 6 R 7 , NR 9 COR 11 , NR 9 CONR 9 ′R 11 , where R 6 , R 7 , R 9 , R 9 ′, and R 11  are each as previously defined, SO(C 1-6 alkyl), SO 2 (C 1-6 alkyl), trifluoromethyl, CONHSO 2 R 8  or SO 2 NHCOR 8 , where R 8  is C 1-6 alkyl, optionally substituted aryl, 2,2-difluorocyclopropane or trifluoromethyl, SO 2 NHR 10 , where R 10  is a nitrogen containing heterocycle, B(OH) 2 , (CH 2 ) q CO 2 H, where q is as previously defined; or  
 R 12  represents a group:  
                     
 where X 1  represents CH or N; W represents CH 2  or NR 9 , where R 9  is as previously defined, and W 1  represents CH 2 , or W and W 1  each represent O; or  
 
         R 12  represents phenyl substituted by a group:  
         
           
             
             
                 
                 
             
           
         
         wherein X 2  is O, S, or NR 9 , where R 9  is as previously defined; z is a bond, O, or S; m is 1, 2 or 3; n is 1, 2, or 3; and y is 0, 1, 2, or 3;  
         R 13  represents a group:  
         
           
             
             
                 
                 
             
           
         
         where R 14  represents H or C 1-6 alkyl; R 15  represents H, C 1-6 alkyl, halo or NR 6 R 7 , where R 6  and R 7  are as previously defined; and the dotted line represents an optional covalent bond; and pharmaceutically acceptable salts or prodrugs thereof,  
         with the provision that, when NR 4 R 5  represents an unsubstituted azacyclic ring system, R 2  does not represent NHR 12  where R 12  is optionally substituted phenyl or:  
         
           
             
             
                 
                 
             
           
         
         where W and W 1  are as previously defined.  
       
     
     
         10 . The method of  claim 9 , wherein said compound is 3(R,S)-Amino-1,3-dihydro-5-((1S,4S)-5-methyl-2,5-diazabicyclo[2,2,1]heptan-2-vl)-2H-1-propyl-1,4-benzodiazepin-2-one.  
     
     
         11 . The method of  claim 9 , wherein said compound is (−)-N-[5-(3-azabicyclo[3,2,2]nonan-3-yl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[3-methylphenyl]-urea.  
     
     
         12 . The method of  claim 9 , wherein said compound is (+)-N-[5-(3-azabicyclo[3,2,2]nonan-3-yl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[3-methylphenyl]-urea.