Azaindazole compounds and methods of use

Abstract

Compounds are provided that act as potent antagonists of the CCR1 receptor, and have in vivo anti-inflammatory activity. The compounds are generally aryl piperazine derivatives and are useful in pharmaceutical compositions, methods for the treatment of CCR1-mediated diseases, and as controls in assays for the identification of competitive CCR1 antagonists.

1 .- 75 . (canceled) 
     
     
         76 . A method of assaying a small organic molecule for CCR1 antagonistic activity, said method comprising
 (a) contacting the small organic molecule with cells expressing CCR1 and a radioactive CCR1 ligand to form a reaction mixture;   (b) aspirating the reaction mixture onto a GF/B glass filter pre-soaked in a 0.3% polyethyleneimine solution;   (c) measuring the amount radioactivity remaining on the GF/B glass filter,
 wherein said method comprises performing steps (a)-(d) with a positive control sample having a formula selected from the group consisting of 
   
       
         
           
           
               
               
           
         
         
           or a pharmaceutically acceptable salt, hydrate or N-oxide thereof, wherein the subscript m is an integer of from 0 to 2; 
         
         each R 1  is independently selected from the group consisting of —CO 2 H and C 1-4  alkyl, optionally substituted with —OH, —OR m , —S(O) 2 R m , —CO 2 H or —CO 2 R m  wherein R m  is an unsubstituted C 1-6  alkyl; 
         R 2a , R 2c  and R 2d  are each independently selected from the group consisting of hydrogen, halogen, cyano, oxazolyl, —NO 2 , —CO 2 R c , —CONR c R d , —C(O)R c , —S(O) 2 R e , —R e , —C(NOR c )R d , —OR c , —SR c , —NR d C(O)R c , —X 2 OR c , —O—X 2 OR c , —X 2 NR c R d , —NR c S(O) 2 R e , —OC(O)R c , and —NR c R d ; wherein 
         within each of R 2a , R 2c  and R 2d , X 2  is C 1-4  alkylene and each R c  and R d  is independently selected from hydrogen, C 1-8  alkyl, C 1-8  haloalkyl, and C 3-6  cycloalkyl and each R e  is independently selected from the group consisting of C 1-8  alkyl, C 1-8  haloalkyl, and C 3-6  cycloalkyl; 
         each of ring vertices a, b, c and d in formulae Ia and Ib is independently selected from N and CH, and one of said ring vertices is N; and 
         R 3a  is selected from the group consisting of hydrogen, halogen, —NR f R g , —R h , —S(O) 2 R h , and —Y, wherein Y is selected from the group consisting of homopiperidinyl, morpholinyl, thiomorpholinyl, pyrrolidinyl, piperidinyl, azetidinyl, pyranyl, tetrahydrofuranyl, piperazinyl, phenyl, thienyl, furanyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, isothiazolyl, triazolyl, tetrazolyl and oxadiazolyl, optionally substituted with from one to three substituents selected from the group consisting of halogen, —OR f , —NR f R g , —R h , —CN, —NO 2 , —CO 2 R f , —CONR f R g , and —C(O)R f , wherein each R f  and R g  is independently selected from hydrogen, C 1-8  alkyl, C 1-8  haloalkyl, and C 3-6  cycloalkyl, and each R h  is independently selected from the group consisting of C 1-8  alkyl, C 1-8  haloalkyl, and C 3-6  cycloalkyl. 
       
     
     
         77 . The method of  claim 76 , wherein said cells expressing CCR1 are THP-1 cells or isolated human monocytes. 
     
     
         78 . The method of  claim 76 , wherein said radioactive CCR1 ligand is MIP-1α, MPIF-1, Leukotactin, or a combination thereof. 
     
     
         79 . The method of  claim 76 , wherein said wash buffer comprises 25 mM Hepes, 500 mM NaCl, 1 mM CaCl 2 , 5 mM MgCl 2 , pH 7.1. 
     
     
         80 . The method of  claim 76 , wherein said measuring comprises adding scintillation fluid to the aspirated and washed GF/C glass filter. 
     
     
         81 . The method of  claim 76 , wherein R 3a  is a member selected from the group consisting of homopiperidinyl, morpholinyl, thiomorpholinyl, pyrrolidinyl, piperidinyl, azetidinyl, pyranyl, tetrahydrofuranyl and piperazinyl. 
     
     
         82 . The method of  claim 76 , wherein R 3a  is a member selected from the group consisting of oxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, triazolyl and thiazolyl. 
     
     
         83 . The method of  claim 76 , wherein m is 0. 
     
     
         84 . The method of  claim 76 , wherein m is 1. 
     
     
         85 . The method of  claim 83 , having formula Ia. 
     
     
         86 . The method of  claim 83 , having formula Ib. 
     
     
         87 . The method of  claim 76 , wherein R 3a  is selected from the group consisting of phenyl, pyridyl, pyrimidinyl, and pyrazinyl. 
     
     
         88 . The method of  claim 76 , wherein m is 0 or 1; and R 2a  is hydrogen. 
     
     
         89 . The method of  claim 76 , wherein R 2a  is selected from the group consisting of hydrogen, F, Cl, Br and I.