Antibody conjugates comprising toll-like receptor agonist

Abstract

Provided herein are antibody conjugates comprising toll-like receptor agonists and the use of such conjugates for the treatment of cancer. In some embodiments, the conjugates comprise anti-HER2 antibodies.

1 . A conjugate of Formula (II), or pharmaceutically acceptable salt thereof: 
       
         
           
           
               
               
           
         
         wherein: 
         R 50  is 
       
       
         
           
           
               
               
           
         
       
       where the * indicates the point of attachment to Ab;
 Ab is an antibody or antigen binding fragment thereof; 
 R 1  is —NHR 2  or —NHCHR 2 R 3 ; 
 R 2  is —C 3 -C 6 alkyl or —C 4 -C 6 alkyl; 
 R 3  is L 1 OH; 
 L 1  is —(CH 2 ) m —; 
 L 2  is —(CH 2 ) n —, —((CH 2 ) n O) t (CH 2 ) n —, —(CH 2 ) n X 1 (CH 2 ) n —, —(CH 2 ) n NHC(═O)(CH 2 ) n —, —(CH 2 ) n NHC(═O)(CH 2 ) n C(═O)NH(CH 2 ) n —, —((CH 2 ) n O) t (CH 2 ) n NHC(═O)(CH 2 ) n , —C(═O)(CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n X 1 (CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n NHC(═O)(CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n C(═O)N H(CH 2 ) n —, —C(═O)NH((CH 2 ) n O) t (CH 2 ) n X 1 (CH 2 ) n —, —C(═O)X 2 X 3 C(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)X 2 X 3 C(═O)(CH 2 ) n —, —C(═O)X 2 C(═O)(CH 2 ) n NHC(═O)(CH 2 ) n —, —C(═O)X 2 C(═O)(CH 2 ) n NHC(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)(CH 2 ) n C(R 7 ) 2 —, —C(═O)(CH 2 ) n C(R 7 ) 2 SS(CH 2 ) n NHC(═O)(CH 2 ) n —, —(CH 2 ) n X 2 C(═O)(CH 2 ) n NHC(═O)((CH 2 ) n O) t (CH 2 ) n — or —C(═O)(CH 2 ) n C(═O)NH(CH 2 ) n ; 
 R 40  is 
 
       
         
           
           
               
               
           
         
       
       —S—, —NHC(═O)CH 2 —, —S(═O) 2 CH 2 CH 2 —, —(CH 2 ) 2 S(═O) 2 CH 2 CH 2 —, —NHS(═O) 2 CH 2 CH 2 , —NHC(═O)CH 2 CH 2 —, —CH 2 NHCH 2 CH 2 —, —NHCH 2 CH 2 —, 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         X 1  is 
       
       
         
           
           
               
               
           
         
         X 2  is 
       
       
         
           
           
               
               
           
         
         X 3  is 
       
       
         
           
           
               
               
           
         
         each R 7  is independently selected from H and C 1 -C 6 alkyl; 
         each R 8  is independently selected from H, C 1 -C 6 alkyl, F, Cl, and —OH; 
         each R 9  is independently selected from H, C 1 -C 6 alkyl, F, Cl, —NH 2 , —OCH 3 , —OCH 2 CH 3 , —N(CH 3 ) 2 , —CN, —NO 2  and —OH; 
         each R 10  is independently selected from H, C 1-6 alkyl, fluoro, benzyloxy substituted with —C(═O)OH, benzyl substituted with —C(═O)OH, C 1-4 alkoxy substituted with —C(═O)OH and C 1-4 alkyl substituted with —C(═O)OH; 
         R 12  is H, methyl or phenyl; 
         each m is independently selected from 1, 2, 3, and 4; 
         each n is independently selected from 1, 2, 3, and 4; 
         each t is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 and 18, and 
         y is an integer from 1 to 16. 
       
     
     
         2 - 3 . (canceled) 
     
     
         4 . The conjugate of  claim 1 , wherein Ab is a human or humanized antibody. 
     
     
         5 . The conjugate of  claim 1 , wherein Ab comprises a modified Fc region. 
     
     
         6 . The conjugate of  claim 1 , wherein Ab comprises cysteine at one or more of the following positions (all positions by EU numbering):
 (a) positions 152, 360 and 375 of the antibody heavy chain, and   (b) positions 107, 159, and 165 of the antibody light chain.   
     
     
         7 . The conjugate of  claim 1 , wherein Ab comprises cysteines at positions 152 and 375 of the antibody heavy chains (all positions by EU numbering). 
     
     
         8 . The conjugate of  claim 1 , wherein the antibody conjugate of Formula (II) comprises the structure of Formula (IIa) or Formula (IIb): 
       
         
           
           
               
               
           
         
       
       wherein:
 R 1  is —NHR 2 ; 
 R 2  is —C 4 -C 6 alkyl; 
 L 2  is —(CH 2 ) n —, —((CH 2 ) n O) t (CH 2 ) n —, —(CH 2 ) n X 1 (CH 2 ) n —, —C(═O)(CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n —, —C(═O)((CH 2 ) n O) t (CH 2 ) n X 1 (CH 2 ) n —, —C(═O)NH((CH 2 ) n O) t (CH 2 ) n X 1 (CH 2 ) n —, —C(═O)X 2 X 3 C(═O)((CH 2 ) n O) t (CH 2 ) n — or —C(═O)X 2 C(═O)(CH 2 ) n NHC(═O)(CH 2 ) n —; 
 R 40  is 
 
       
         
           
           
               
               
           
         
         X 1  is 
       
       
         
           
           
               
               
           
         
       
       X 2  is 
       
         
           
           
               
               
           
         
       
       X 3  is 
       
         
           
           
               
               
           
         
         each n is independently selected from 1, 2, 3, and 4; 
         each t is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 and 18, and 
         y is an integer from 1 to 16. 
       
     
     
         9 . The conjugate of  claim 1 , wherein
 R 1  is —NHR 2 ;   R 2  is —C 4 -C 6 alkyl;   L 2  is —(CH 2 ) n — or —C(═O)(CH 2 ) n ;   R 40  is   
       
         
           
           
               
               
           
         
         and 
         each n is independently selected from 1, 2, 3, and 4, and 
         y is an integer from 1 to 16. 
       
     
     
         10 . The conjugate of  claim 1 , wherein the conjugate has a hydrophobicity index of 0.8 or greater, as determined by hydrophobic interaction chromatography. 
     
     
         11 . A conjugate comprising any of the following formulas: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       wherein Ab is an antibody or antigen binding fragment thereof, and y is an integer from 1 to 4. 
     
     
         12 - 14 . (canceled) 
     
     
         15 . The conjugate of  claim 11 , wherein the compound is attached to cysteines at positions 152 and 375 of the antibody heavy chain (all positions by EU numbering). 
     
     
         16 . The conjugate of  claim 11 , wherein y is about 3 to 4. 
     
     
         17 . The conjugate of  claim 11 , wherein the conjugate has a hydrophobicity index of 0.8 or greater, as determined by hydrophobic interaction chromatography. 
     
     
         18 . A pharmaceutical composition comprising one or more conjugates of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         19 . A pharmaceutical composition comprising one or more conjugates of  claim 11  and a pharmaceutically acceptable carrier. 
     
     
         20 . A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the conjugate of  claim 1 . 
     
     
         21 . (canceled) 
     
     
         22 . The method of  claim 20 , wherein the cancer is selected from gastric cancer, esophageal cancer, gastroesophageal junction adenocarcinoma, colon cancer, rectal cancer, breast cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, bladder cancer, urinary tract cancer, pancreatic cancer, lung cancer, prostate cancer, osteosarcoma, neuroblastoma, glioblastoma, and head and neck cancer. 
     
     
         23 . The method of  claim 20 , wherein the conjugate is administered to the subject intravenously, intratumorally, or subcutaneously. 
     
     
         24 . The method of  claim 20 , wherein the conjugate is administered at a dose of about 0.01-20 mg per kg of body weight. 
     
     
         25 . The method of  claim 20  further comprising administering a second agent to the subject. 
     
     
         26 - 33 . (canceled)