Antagonists of chemokine receptors

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 .- 27 . (canceled) 
     
     
         28 . A method of assaying a compound for CCR1 antagonistic activity, said method comprising
 (a) contacting the compound 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 polyethyleneimine solution;   (c) measuring the amount radioactivity remaining on the GF/B glass filter,
 wherein said method comprises performing steps (a)-(c) with a positive control sample having a formula represented by the structure 
   
       
         
           
           
               
               
           
         
       
       wherein
 the subscript n is an integer of from 0 to 3; 
 each R 1a  and R b  is a member independently selected from the group consisting of H, C 1-8  alkyl, C 1-8  haloalkyl, C 3-6  cycloalkyl, —COR a , —CO 2 R a , —CONR a R b , —NR a R b , —NR a COR b , —OR a , —X 1 COR a , —X 1 CO 2 R a , —X 1 CONR a R b , —X 1 NR a COR b , —X 1 NR a R b , and —X 1 OR a , wherein X 1  is a member selected from the group consisting of C 1-4  alkylene, and each R a  and R b  is independently selected from the group consisting of hydrogen, C 1-8  alkyl, C 1-8  haloalkyl, and C 3-6  cycloalkyl, and optionally two R 1a  groups on adjacent carbon atoms are joined to form a 5-, 6- or 7-membered carbocyclic or heterocyclic ring; 
 each of R 2a  and R 2b  is a member independently selected from the group consisting of H, hydroxyl, C 1-8  alkyl, C 1-8  haloalkyl, C 1-8  alkoxy, C 1-4  alkoxy-C 1-4  alkyl, C 1-8  hydroxyalkyl, C 1-4  alkoxy-C 1-4  alkoxy, C 3-6  cycloalkyl, C 3-6  cycloalkyl-C 1-4  alkyl, 3- to 7-membered heterocycloalkyl, 3- to 7-membered heterocycloalkyl-C 1-4  alkyl, —X 1 CO 2 R a , —X 1 CONR a R b , —X 1 NR a COR b , —X 1 NR a R b , wherein X 1 , R a  and R b  are defined above; 
 Ar 1  is a member selected from the group consisting of phenyl, naphtyl and pyridyl, each of which is substituted with from one to five substituents, R 3 , R 3a , R 3b , R 4  and R 4a  which are independently selected from the group consisting of H, halogen, —OR c , —OC(O)R c , —NR c R d , —SR c , —R c , —CN, —NO 2 , —CO 2 R c , —CONR c R d , —C(O)R c , —OC(O)NR c R d , —NR d C(O)R c , —NR d C(O) 2 R, —NR c —C(O)NR c R d , —NH—C(NH 2 )═NH, —NR e C(NH 2 )═NH, —NH—C(NH 2 )═NR e , —NH—C(NHR e )═NH, —S(O)R, —S(O) 2 R e , —NR c S(O) 2 R e , —S(O) 2 NR c R d , —N 3 , —X 2 OR c , —O—X 2 OR c , —X 2 OC(O)R c , —X 2 NR c R d , —O—X 2 NR c R d , —X 2 SR c , —X 2 CN, —X 2 NO 2 , —X 2 CO 2 R c , —O—X 2 CO 2 R c , —X 2 CONR c R d , —O—X 2 CONR c R d , —X 2 C(O)R c , —X 2 OC(O)NR c R d , —X 2 NR d C(O)R c , —X 2 NR d C(O) 2 R e , —X 2 NR c C(O)NR c R d , —X 2 NH—C(NH 2 )═NH, —X 2 NR e C(NH 2 )═NH, —X 2 NH—C(NH 2 )═NR e , —X 2 NH—C(NHR e )═NH, —X 2 S(O)R, —X 2 S(O) 2 R, —X 2 NR c S(O) 2 R e , —X 2 S(O) 2 NR c R d , —X 2 N 3 , —NR d —X 2 OR, —NR d —X 2 NR c R d , —NR d —X 2 CO 2 R c , and —NR d —X 2 CONR c R d , wherein each X 2  is a member independently selected from the group consisting of C 1-4  alkylene, and each R c  and R d  is independently selected from hydrogen, C 1-8  alkyl, C 1-8  hydroxyalkyl, C 1-8  haloalkyl and C 3-6  cycloalkyl, or optionally R and R d  when attached to the same nitrogen atom can be combined with the nitrogen atom to form a five or six-membered ring having from 0 to 2 additional heteroatoms as ring members; and each R is independently selected from the group consisting of C 1-8  is alkyl, C 1-8  hydroxyalkyl, C 1-8  haloalkyl and C 3-6  cycloalkyl; 
 Ar 2  is a member selected from the group consisting of a six- or ten-membered monocyclic or fused bicyclic aryl ring, and a five- to ten-membered monocyclic or fused bicyclic heteroaryl ring; each of which is substituted with from one to five substituents, R 5 , R 6 , R 7 , R 8  and R 9 , independently selected from the group consisting of H, halogen, —OR f , —OC(O)R f , —NR f R g , —SR, —R h , —CN, —NO 2 , —CO 2 R f , —CONR f R g , —C(O)R f , —OC(O)NR f R g , —NR g C(O)R f , —NR g C(O) 2 R h , —NR f —C(O)NR f R g , —NH—C(NH 2 )═NH, —NR h C(NH 2 )═NH, —NH—C(NH 2 )═NR h , —NH—C(NHR h )═NH, —S(O)R h , —S(O) 2 R h , —NR f S(O) 2 R h , —S(O) 2 NR f R g , —NR f S(O) 2 NR f R g , —N 3 , —X 3 OR, —X 3 OC(O)R f , —X 3 NR f R g , —X 3 SR f , —X 3 CN, —X 3 NO 2 , —X 3 CO 2 R, —X 3 CONR f R g , —X 3 C(O)R f , —X 3 OC(O)NR f R g , —X 3 NR g C(O)R f , —X 3 NR g C(O) 2 R h , —X 3 NR f —C(O)NR f R g , —X 3 NH—C(NH 2 )═NH, —X 3 NR h C(NH 2 )═NH, —X 3 NH—C(NH 2 )═NR h , —X 3 NH—C(NHR h )═NH, —X 3 S(O)R h , —X 3 S(O) 2 R h , —X 3 NR f S(O) 2 R h , —X 3 S(O) 2 NR f R g , —Y, —X 3 Y, —S(O) 2 Y, —C(O)Y, —X 3 N 3 , —O—X 3 OR f , —O—X 3 NR f R g , —O—X 3 CO 2 R, —O—X 3 CONR f R g , —NR g —X 3 OR f , —NR g —X 3 NR f R g , —NR g —X 3 CO 2 R f , and —NR 9 —X 3 CONR f R g , wherein Y is a five or six-membered aryl, heteroaryl or heterocyclic ring, optionally substituted with from one to three substitutents selected from the group consisting of halogen, —OR f , —OC(O)R, —NR f R g , —R h , —SR f , —CN, —NO 2 , —CO 2 R f , —CONR f R g , —C(O)R f , —NR g C(O)R f , —NR g C(O) 2 R h , —S(O)R h , —S(O) 2 R h , —NR f S(O) 2 R h , —S(O) 2 NR f R g , —X 3 OR f , —X 3 SR f , —X 3 CN, —X 3 NO 2 , —X 3 CO 2 R f , —X 3 CONR f R g , —X 3 C(O)R f , —X 3 OC(O)NR f R g , —X 3 NR g C(O)R f , —X 3 NR g C(O) 2 R h , —X 3 NR f —C(O)NR f R g , —X 3 OC(O)R f , —X 3 S(O)R h , —X 3 S(O) 2 R h , —X 3 NR f R g , —X 3 NR f S(O) 2 R h , —X 3 S(O) 2 NR f R g , —O—X 3 OR f , —O—X 3 NR f R g , —O—X 3 CO 2 R, —O—X 3 CONR f R g , —NR g —X 3 OR f , —NR g —X 3 NR f R g , —NR g —X 3 CO 2 R f , and —NR g —X 3 CONR f R g  and wherein each X 3  is independently selected from the group consisting of C 1-4  alkylene, and each R f  and R g  is independently selected from hydrogen, C 1-8  alkyl, C 1-8  hydroxyalkyl, C 1-8  haloalkyl and C 3-6  cycloalkyl, or when attached to the same nitrogen atom can be combined with the nitrogen atom to form a five or six-membered ring having from 0 to 2 additional heteroatoms as ring members, and each R h  is independently selected from the group consisting of C 1-8  alkyl, C 2-8  alkenyl, C 2-8  alkynyl, C 1-8  hydroxyalkyl, C 1-8  haloalkyl and C 3-6  cycloalkyl;
 or when two of R 5 , R 6 , R 7 , R 8  and R 9 , are attached to adjacent ring vertices of Ar 2 , are optionally combined to form a five or six membered ring having zero, one or two heteroatoms selected from O and N as ring members; 
 or a pharmaceutically acceptable salt, or rotamer thereof. 
 
 
     
     
         29 . The method of  claim 28 , wherein said cells expressing CCR1 are THP-1 cells or isolated human monocytes. 
     
     
         30 . The method of  claim 28 , wherein said radioactive CCR1 ligand is MIP-1α, MPIF-1, Leukotactin, or a combination thereof. 
     
     
         31 . The method of  claim 28 , wherein said reaction mixture comprises an assay buffer comprising 25 mM Hepes, 500 mM NaCl, 1 mM CaCl 2 , 5 mM MgCl 2 , pH 7.1. 
     
     
         32 . The method of  claim 28 , wherein said measuring comprises adding scintillation fluid to the aspirated GF/C glass filter. 
     
     
         33 . The method of  claim 28 , wherein Ar 2  is selected from the group consisting of phenyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, oxathiadiazolyl, pyrrolyl, thiazolyl, isothiazolyl, benzimidazolyl, benzoxazolyl, benzopyrazolyl, benzotriazolyl, pyrazolo[3,4-b]pyridine, pyrazolo[3,4-d]pyrimidine, imidazo[4,5-b]pyridine, imidazo[1,5-α]pyridine, and pyrrolo[2,3-b]pyridine, each of which is optionally substituted with R 5 , R 6  and R 7 . 
     
     
         34 . The method of  claim 28 , wherein Ar 2  is selected from the group consisting of pyrazolyl, imidazolyl and triazolyl, each of which is substituted with R 5 , R 6  and R 7 . 
     
     
         35 . The method of  claim 28 , Ar 1  is selected from the group consisting of phenyl, naphthyl and pyridyl, each of which is substituted with from one to five substituents, R 3 , R 3a , R 3b , R 4  and R 4a ; and Ar 2  is selected from the group consisting of pyrazolyl, imidazolyl and triazolyl, each of which is substituted with R 5 , R 6  and R 7 . 
     
     
         36 . The method of  claim 28 , wherein Ar 1  is phenyl, which is substituted with from one to five substituents, R 3 , R 3a , R 3b , R 4  and R 4a , and Ar 2  is selected from the group consisting of pyrazolyl, imidazolyl, benzimidazolyl, benzopyrazolyl, pyrazolo[3,4-b]pyridine, pyrazolo[3,4-d]pyrimidine, imidazo[4,5-b]pyridine, imidazo[1,5-α]pyridine, and pyrrolo[2,3-b]pyridine, each of which is optionally substituted with R 5 , R 6  and R 7 . 
     
     
         37 . The method of  claim 28 , wherein the positive control sample has a formula represented by the structure: 
       
         
           
           
               
               
           
         
         wherein R 3  and R 4  are independently selected from the group consisting of H, halogen, —R e , —CN, and —SO 2 R. 
       
     
     
         38 . The method of  claim 37 , wherein Ar 2  is a heteroaryl group. 
     
     
         39 . The method of  claim 37 , wherein Ar 2  is a heteroaryl group, optionally substituted and attached to the remainder of the molecule through a nitrogen atom ring vertex. 
     
     
         40 . The method of  claim 39 , wherein said Ar 2  has the formula: 
       
         
           
           
               
               
           
         
         wherein R 5 , R 6 , and R 7  are independently selected from the group consisting of H, halogen, —R h , —CN, —SO 2 R h , —CO 2 R f , —CONR f R g , and Y. 
       
     
     
         41 . The method of  claim 40 , the positive control sample has a formula represented by the structure: 
       
         
           
           
               
               
           
         
         wherein R 4  is selected from the group consisting of F and C 1 . 
       
     
     
         42 . The method of  claim 41 , wherein Y is selected from the group consisting of pyridyl, pyrimidinyl, imidazolyl, oxazolyl, oxadiazolyl, triazolyl, thiazolyl, imidazolinyl and pyrazolyl. 
     
     
         43 . The method of  claim 41 , the positive control sample has a formula represented by the structure: 
       
         
           
           
               
               
           
         
         wherein R 3  is selected from the group consisting of H, halogen, C 1-8  alkyl, C 1-8  haloalkyl and C 1-8  alkoxy; R 1a  and R 2  are independently selected from the group consisting of H, C 1-8  alkyl, C 1-8  haloalkyl, C 1-8  alkoxy and C 1-8  hydroxyalkyl. 
       
     
     
         44 . The method of  claim 28 , the positive control sample has a formula represented by the structure: 
       
         
           
           
               
               
           
         
       
       wherein R 1a  and R 2a  are independently selected from the group consisting of H, C 1-8  alkyl, C 1-8  haloalkyl, C 1-8  alkoxy and C 1-8  hydroxyalkyl; and R 5 , R 6 , and R 7  are independently selected from the group consisting of H, halogen, —R h , —CN, —SO 2 R h , —CO 2 R f —CONR f R g , and Y. 
     
     
         45 . The method of  claim 28 , the positive control sample has a formula represented by the structure: 
       
         
           
           
               
               
           
         
       
       Where R 1a  and R 2a  are independently selected from the group consisting of H, C 1-8  alkyl, C 1-8  haloalkyl, C 1-8  alkoxy and C 1-8  hydroxyalkyl; and R 5 , R 6 , and R 7  are independently selected from the group consisting of H, halogen, —R h , —CN, —SO 2 R h , —CO 2 R f —CONR f R g , and Y. 
     
     
         46 . The method of  claim 28 , the positive control sample has a formula represented by the structure: 
       
         
           
           
               
               
           
         
         wherein R 1a  and R 2a  are each independently selected from the group consisting of H, C 1-8  alkyl, C 1-8  haloalkyl, C 1-8  alkoxy, and C 1-8  hydroxyalkyl; and R 5  and R 7  are each independently selected from the group consisting of H, halogen, —R h , —CN, —SO 2 R h , —CO 2 R f , —CONR f R g , and Y. 
       
     
     
         47 . The method of  claim 28 , the positive control sample has a formula selected from the group consisting of