Anti-asgr1 antibody conjugates and uses thereof

Abstract

The disclosure provides conjugates of anti-ASGR1 antibodies or antigen binding fragments thereof to a myeloid cell agonist, compositions comprising the conjugates, and methods of treating liver viral infections with the conjugates. The disclosure also provides for anti-ASGR1 antibodies or antigen binding fragments thereof and methods for using the antibodies or antigen binding fragments thereof in treating liver viral infections.

1 . A method of treating a subject having a liver viral infection, comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising:
 a myeloid cell agonist conjugate or a salt thereof, wherein the conjugate is represented by Formula (I):   
       
         
           
           
               
               
           
         
       
       wherein:
 A is an anti-ASGR1 antibody or antigen-binding fragment thereof, comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein: 
 the VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of any one of SEQ ID NOS:1, 2, 252-255, and 266-269, a VH-CDR2 comprising the amino acid sequence selected from any one of SEQ ID NOS:6-9, 256-259, and 270-273, a VH-CDR3 comprising the amino acid sequence of any one of SEQ ID NOS:13, 14, 260, 261, 274, and 275; and the VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of any one of SEQ ID NOS:18, 19, 262, 263, and 276, a VL-CDR2 comprising the amino acid sequence selected from any one of SEQ ID NOS:23-27, 264, 277, and “RA”, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO:33, 34, 265, 278, and 279; 
 L is a linker; 
 D x  comprises a TLR8 agonist; 
 n is selected from 1 to 20; and 
 z is selected from 1 to 20; 
 and a pharmaceutically acceptable excipient. 
 
     
     
         2 . The method of  claim 1 , wherein the anti-ASGR1 antibody or antigen-binding fragment thereof comprises:
 (a) a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:240, and a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:247; or   (b) a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:239, and a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:247; or   (c) a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:239, and a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:248; or   (d) a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:240, and a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:248; or   (e) a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:43, and a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:131; or   (f) a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:89, and a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:134; or   (g) a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:89, and a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:136; or   (h) a VH comprising the amino acid sequence of SEQ ID NO:239, and a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:247; or   (i) a VH comprising the amino acid sequence of SEQ ID NO:240, and a VL comprising the amino acid sequence of SEQ ID NO:247; or   (j) a VH comprising the amino acid sequence of SEQ ID NO:239, and a VL comprising the amino acid sequence of SEQ ID NO:247; or   (k) a VH comprising the amino acid sequence of SEQ ID NO:239, and a VL comprising the amino acid sequence of SEQ ID NO:248; or   (l) a VH comprising the amino acid sequence of SEQ ID NO:240, and a VL comprising the amino acid sequence of SEQ ID NO:248; or   (m) a VH comprising the amino acid sequence of SEQ ID NO:89, and a VL comprising the amino acid sequence of SEQ ID NO:134; or   (n) a VH comprising the amino acid sequence of SEQ ID NO:89, and a VL comprising the amino acid sequence of SEQ ID NO:136.   
     
     
         3 . The method of  claim 1 , wherein A is an anti-ASGR1 antibody and the antibody comprises:
 (a) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:240 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (b) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:239 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (c) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:239 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:248 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (d) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:240 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:248 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (e) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:89 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:134 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (f) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:89 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:136 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (g) a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:240 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232;   (h) a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:239 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232; or   (i) a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:239 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:248 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232; or   (j) a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:240 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:248 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232; or   (k) a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:89 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:134 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232; or   (l) a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:89 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:136 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232.   
     
     
         4 . The method of  claim 1 , wherein the antibody comprises a human IgG1 constant region. 
     
     
         5 . The method of  claim 1 , wherein the TLR8 agonist is selected from compounds 1.1-1.69 or a salt thereof. 
     
     
         6 . The method of  claim 1 , wherein the TLR8 agonist is compound 1.36, 1.50, 1.57, 1.60, or 1.64 or a salt thereof. 
     
     
         7 . The method of  claim 1 , wherein the TLR8 agonist is 
       
         
           
           
               
               
           
         
       
     
     
         8 . The method of  claim 1 , wherein the linker is represented by formula (V): 
       
         
           
           
               
               
           
         
         wherein L 4  represents the C-terminal of the peptide and L 5  is selected from a bond, alkylene and heteroalkylene, wherein L 5  is optionally substituted with one or more groups independently selected from R 32 ; RX* comprises a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of the antibody, wherein   on RX* represents the point of attachment to the residue of the antibody and the other   represents the point of attachment to the TLR8 agonist; and R 32  is independently selected at each occurrence from halogen, —OH, —CN, —O— C 1-10  alkyl, —SH, ═O, ═S, —NH 2 , —NO 2 ; C 1-10  alkyl, C 2-10  alkenyl, and C 2-10  alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen, —OH, —CN, —O—C 1-10  alkyl, —SH, ═O, ═S, —NH 2 , and —NO 2 . 
       
     
     
         9 . The method of  claim 8 , wherein the linker is a cleavable linker. 
     
     
         10 . The method of  claim 8 , wherein the peptide of the linker is Val-Cit or Val-Ala. 
     
     
         11 . The method of  claim 1 , wherein L-D x  has a structure selected from any one of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
       and a salt of any one thereof,
 wherein the RX* is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of the antibody, and 
 wherein   on RX* represents the point of attachment to the residue of the antibody. 
 
     
     
         12 . The method of  claim 11 , wherein
 (a) n is an integer from 1 to about 10, or from 1 to about 5, or is 1 or 2, or is 1; and   (b) z ranges from 1 to about 10, or from 1 to about 9, or from 1 to about 8, or from 2 to about 6, or from 3 to about 5, or is about 4.   
     
     
         13 . The method of  claim 1 , wherein the average TLR8 agonist-to-antibody ratio of the myeloid cell agonist conjugate ranges from about 2 to about 8, from about 3 to about 8, from about 3 to about 7, or from about 3 to about 5. 
     
     
         14 . The method of  claim 1 , wherein the average TLR8 agonist-to-antibody ratio of the myeloid cell agonist conjugate is from 2 to 4. 
     
     
         15 . The method of  claim 1 , wherein:
 A is an anti-ASGR1 antibody comprising:   (a) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:240 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (b) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:239 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (c) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:239 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:248 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (d) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:240 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:248 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (e) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:89 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:134 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232; or   (f) a heavy chain comprising a VH comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:89 and a human IgG1 constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:136 and a human kappa constant region comprising the amino acid sequence that has at least 90% identity with the amino acid sequence of SEQ ID NO:232;   L-D x  has structure selected from any one of:   
       
         
           
           
               
               
           
         
       
       wherein RX* is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety bound to a residue of the antibody, wherein   RX* represents the point of attachment to a cysteine residue of the antibody;
 n is 1; and 
 z ranges from 2 to about 6. 
 
     
     
         16 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:239 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         17 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:239 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         18 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:239 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         19 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:239 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         20 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:240 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         21 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:240 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         22 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:240 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         23 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:240 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:247 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         24 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:89 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:134 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         25 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:89 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:134 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         26 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:89 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO: 134 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         27 . The method of  claim 15 , wherein A is an anti-ASGR1 antibody comprising comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO:89 and a human IgG1 constant region comprising the amino acid sequence of SEQ ID NO:230, and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO:134 and a human kappa constant region comprising the amino acid sequence of SEQ ID NO:232, and L-D x  has a structure of: 
       
         
           
           
               
               
           
         
       
     
     
         28 . The method of  claim 14 , wherein the liver viral infection is Hepatitis B or Hepatitis C. 
     
     
         29 . The method of  claim 14 , wherein the subject does not have cancer. 
     
     
         30 . The method of  claim 14 , wherein the pharmaceutical composition is administered systemically. 
     
     
         31 . The method of  claim 14 , wherein the pharmaceutical composition is administered intravenously, cutaneously, subcutaneously, or injected at a site of the liver viral infection. 
     
     
         32 . The method of  claim 14 , wherein the pharmaceutical composition is administered intravenously. 
     
     
         33 . The method of  claim 14 , wherein the pharmaceutical composition is administered subcutaneously. 
     
     
         34 . The method of  claim 14 , wherein the method comprises administering an additional therapeutic agent. 
     
     
         35 . The method of  claim 34 , wherein the additional therapeutic agent comprises an anti-viral drug, a vaccine, an immunomodulatory drug, or any combination thereof. 
     
     
         36 . The method of  claim 35 , wherein the immunomodulatory drug comprises a TLR agonist, a RIG-I agonist, an interferon therapy, an anti-PD1 therapy, an anti-PD-L1 therapy, a PD-L1 inhibitor, or any combination thereof. 
     
     
         37 . The method of  claim 35 , wherein the immunomodulatory drug comprises an anti-PD1 therapy, an anti-PD-L1 therapy, a PD-L1 inhibitor, or any combination thereof. 
     
     
         38 . The method of  claim 35 , wherein the immunomodulatory drug comprises Interferon α-2b (Intron A), peginterferon α-2a (Pegasys), thymalfasin (Zadaxin), Inarigivir, nivolumab, pembrolizumab, pidilizumab, toripalimab, cemiplimab, dostarlimab, spartalizumab, camrelizumab, sintilimab, tislelizumab, atezolizumab, GS-4224, RG6084 (RO7191863), avelumab, durvalumab, vesatolimod, AZD8848, MEDI9197, ISA204, RG7854 (RO7020531), JNJ-4964 (AL-034), aurigenel, BMSpep-57, BMS-103, BMS-142, BMS-1166, ASC22, selgantolimod, motolimod, a TLR7 agonist, a TLR8 agonist, or any combination thereof.