Anti-cxcr3 antibodies and methods of use thereof

Abstract

The present disclosure provides anti-CXCR3 antibodies and methods of using the antibodies to diagnose and/or treat CXCR3-associated disorders such as diabetes mellitus type I (T1D), particularly new-onset T1D. In certain embodiments, disclosed herein are CXCR3 neutralizing antibodies.

What is claimed is: 
     
         1 . An isolated nucleic acid encoding the amino acid sequence of an antibody or antigen binding fragment capable of binding to Chemokine (C-X-C motif) receptor 3 (CXCR3), said antibody or antigen binding fragment comprising six complementarity determining regions (CDRs): heavy chain variable domain (VH) CDR1, VH CDR2, VH CDR3, light chain variable domain (VL) CDR1, VL CDR2, and VL CDR3,
 wherein:   VH CDR1 is selected from the group consisting of:   
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 116) 
                 
                     
                   GISFNDAA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 172) 
                 
                     
                   GFTFTSYA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 228) 
                 
                     
                   GFTFSNYA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 368) 
                 
                     
                   GFTFTSYA, 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 543) 
                 
                     
                   GYTFTDYA; 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         VH CDR2 is selected from the group consisting of: 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 118) 
                 
                     
                   IRSKINDYGT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 174) 
                 
                     
                   ISHGGSYT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 230) 
                 
                     
                   ISNGGSYT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 370) 
                 
                     
                   ISHGGTYT, 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 545) 
                 
                     
                   ISTYNGNT, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         VH CDR3 is selected from the group consisting of: 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 120) 
                 
                     
                   VIDGYGSLAY, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 176) 
                 
                     
                   ARHPFYSGNYQGYFDY, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 232) 
                 
                     
                   SRPSERSHYYATSQFAY, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 372) 
                 
                     
                   ARHPIYSGNYQGYFDY, 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 547) 
                 
                     
                   ARFLSLRYFDV, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         VL CDR1 is selected from the group consisting of: 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 123) 
                 
                     
                   SSVISSY, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 179) 
                 
                     
                   SGVNY, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 235) 
                 
                     
                   SSVSY, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 375) 
                 
                     
                   SGVNY, 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 550) 
                 
                     
                   SSVIY, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         VL CDR2 is selected from the group consisting of: 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 125) 
                 
                     
                   STS, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 181) 
                 
                     
                   FTS, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 237) 
                 
                     
                   DTS, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 377) 
                 
                     
                   FTS, 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 552) 
                 
                     
                   ATS, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         and 
         VL CDR3 is selected from the group consisting of: 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 127) 
                 
                     
                   QQYSGYPLT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 183) 
                 
                     
                   QQFTSSPYT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 239) 
                 
                     
                   QQWSSSPLT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 379) 
                 
                     
                   QQFTSSPYT, 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 554) 
                 
                     
                   QQWSSEPLT. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         2 . The isolated nucleic acid of  claim 1 , wherein the antibody or fragment is chimeric, CDR grafted, mutated, mutated to remove one or more deamidation site, human, humanized, humanized and back-mutated, synthetic, or recombinant. 
     
     
         3 . The isolated nucleic acid of  claim 1 , wherein the antibody or fragment is capable of binding to a polypeptide comprising a peptide selected from the group consisting of:
 a) a peptide comprising the amino acid sequence SDHQVLNDAE (SEQ ID NO:71);   b) a peptide comprising the amino acid sequence SDHQVLND (SEQ ID NO:72);   c) a peptide comprising the amino acid sequence DHQVLND (SEQ ID NO:73);   d) a peptide comprising the amino acid sequence VLNDAE (SEQ ID NO:74);   e) a peptide comprising the amino acid sequence VLND (SEQ ID NO:75);   f) a peptide comprising the amino acid sequence XDXXVXNDXX (SEQ ID NO:76);   g) a peptide comprising the amino acid sequence XDXXVXND (SEQ ID NO:77);   h) a peptide comprising the amino acid sequence DXXVXND (SEQ ID NO:78);   i) a peptide comprising the amino acid sequence VXNDXX (SEQ ID NO:79);   j) a peptide comprising the amino acid sequence VXND (SEQ ID NO:80);   k) a peptide comprising residues 1-58 of SEQ ID NO:1;   l) a peptide comprising residues 1-16 of SEQ ID NO:1; and   m) a peptide comprising residues 1-37 of SEQ ID NO:1,   wherein X indicates any amino acid.   
     
     
         4 . The isolated nucleic acid of  claim 1 , wherein the antibody or fragment comprises a heavy chain variable region and a light chain variable region,
 wherein the heavy chain variable region comprises a sequence at least about 85%, 90%, 95%, or 99% identical to a sequence selected from the group consisting of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26-33, 38, 40, 42, 44, 46-48, 63-66, 55, 57, 59, and 61;   and   wherein the light chain variable region comprises a sequence at least about 85%, 90%, 95%, or 99% identical to a sequence selected from the group consisting of SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 34-37, 39, 41, 43, 45, 49-54, 67-70, 56, 58, 60, and 62.   
     
     
         5 . The isolated nucleic acid of  claim 4 , wherein the heavy chain variable region comprises a sequence selected from the group consisting of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26-33, 38, 40, 42, 44, 46-48, 63-66, 55, 57, 59, and 61;
 and   wherein the light chain variable region comprises a sequence selected from the group consisting of SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 34-37, 39, 41, 43, 45, 49-54, 67-70, 56, 58, 60, and 62.   
     
     
         6 . The isolated nucleic acid of  claim 1 , wherein said antibody or fragment comprises 3 CDRs selected from the group of variable domain CDR sets consisting of:
 Clone 12 VH CDR set: SEQ ID NO: 116, SEQ ID NO: 118, and SEQ ID NO: 120;   Clone 12 VL CDR set: SEQ ID NO: 123, SEQ ID NO: 125, and SEQ ID NO: 127;   Clone 135 VH CDR set: SEQ ID NO: 172, SEQ ID NO: 174, and SEQ ID NO: 176;   Clone 135 VL CDR set: SEQ ID NO: 179, SEQ ID NO: 181, and SEQ ID NO: 183;   Clone 4 VH CDR set: SEQ ID NO: 228, SEQ ID NO: 230, and SEQ ID NO: 232;   Clone 4 VL CDR set: SEQ ID NO: 235, SEQ ID NO: 237, and SEQ ID NO: 239;   Clone 53 VH CDR set: SEQ ID NO: 368, SEQ ID NO: 370, and SEQ ID NO: 372;   Clone 53 VL CDR set: SEQ ID NO: 375, SEQ ID NO: 377, and SEQ ID NO: 379;   Clone 82 VH CDR set: SEQ ID NO: 543, SEQ ID NO: 545, and SEQ ID NO: 547;   and   Clone 82 VL CDR set: SEQ ID NO: 550, SEQ ID NO: 552, and SEQ ID NO: 554.   
     
     
         7 . The isolated nucleic acid of  claim 1 , wherein said antibody or fragment comprises
 two variable domain CDR sets selected from a group consisting of:   Clone 12 VH CDR set and Clone 12 VL CDR set;   Clone 135 VH CDR set and Clone 135 VL CDR set;   Clone 4 VH CDR set and Clone 4 VL CDR set;   Clone 53 VH CDR set and Clone 53 VL CDR set;   and   Clone 82 VH CDR set and Clone 82 VL CDR set.   
     
     
         8 . The isolated nucleic acid of  claim 1 , wherein the antibody or fragment comprises a combination of heavy chain and light chain variable regions selected from the group consisting of:
 SEQ ID NOs: 18 and 19; SEQ ID NOs: 20 and 21; SEQ ID NOs: 22 and 23; SEQ ID NOs: 24 and 25; SEQ ID NOs: 22 and 25; SEQ ID NOs: 24 and 23; SEQ ID NOs: 26 and 34; SEQ ID NOs: 26 and 37; SEQ ID NOs: 27 and 35; SEQ ID NOs: 27 and 36; SEQ ID NOs: 28 and 34; SEQ ID NOs: 22 and 21; SEQ ID NOs: 20 and 23; SEQ ID NOs: 24 and 21; SEQ ID NOs: 20 and 25; SEQ ID NOs: 29 and 23; SEQ ID NOs: 30 and 23; SEQ ID NOs: 31 and 23; SEQ ID NOs: 32 and 23; SEQ ID NOs: 33 and 23; SEQ ID NOs: 2 and 3; SEQ ID NOs: 4 and 5; SEQ ID NOs: 6 and 7; SEQ ID NOs: 8 and 9; SEQ ID NOs: 55 and 56; SEQ ID NOs: 57 and 58; SEQ ID NOs: 59 and 60; SEQ ID NOs: 61 and 62; SEQ ID NOs: 10 and 11; SEQ ID NOs: 12 and 13; SEQ ID NOs: 14 and 15; SEQ ID NOs: 16 and 17; SEQ ID NOs: 38 and 39; SEQ ID NOs: 40 and 41; SEQ ID NOs: 42 and 43 SEQ ID NOs: 44 and 45; SEQ ID NOs: 40 and 43; SEQ ID NOs: 42 and 41; SEQ ID NOs: 42 and 49; SEQ ID NOs: 42 and 50; SEQ ID NOs: 42 and 51; SEQ ID NOs: 42 and 52; SEQ ID NOs: 42 and 53; SEQ ID NOs: 42 and 54; SEQ ID NOs: 46 and 43; SEQ ID NOs: 47 and 43; SEQ ID NOs: 48 and 43; SEQ ID NOs: 40 and 49; SEQ ID NOs: 40 and 51; SEQ ID NOs: 48 and 49; SEQ ID NOs: 48 and 51; SEQ ID NOs: 63 and 67; SEQ ID NOs: 63 and 68.   
     
     
         9 . The isolated nucleic acid of  claim 1 , wherein the antibody or fragment is capable of preferentially binding to A-isoform of CXCR3. 
     
     
         10 . A vector comprising the isolated nucleic acid of  claim 1 . 
     
     
         11 . A vector comprising the isolated nucleic acid of  claim 7 . 
     
     
         12 . A vector comprising the isolated nucleic acid of  claim 8 . 
     
     
         13 . A host cell comprising the vector of  claim 10 . 
     
     
         14 . A host cell comprising the vector of  claim 11 . 
     
     
         15 . The host cell according to  claim 13 , wherein said host cell is a cell selected from the group consisting of an  E. coli  cell, a COS cell, and a Chinese hamster ovary (CHO) cell. 
     
     
         16 . The host cell according to  claim 14 , wherein said host cell is a cell selected from the group consisting of an  E. coli  cell, a COS cell, and a Chinese hamster ovary (CHO) cell. 
     
     
         17 . A method of producing the antibody or fragment encoded by the isolated nucleic acid of  claim 1 , comprising culturing the host cell of  claim 13  in a culture medium under conditions suitable to produce said antibody or fragment. 
     
     
         18 . A method of producing the antibody or fragment encoded by the isolated nucleic acid of  claim 1 , comprising culturing the host cell of  claim 14  in a culture medium under conditions suitable to produce said antibody or fragment. 
     
     
         19 . The method of  claim 17 , wherein said antibody or fragment is capable of preferentially binding to A-isoform of CXCR3. 
     
     
         20 . The method of  claim 17 , wherein said antibody or fragment is capable of preventing, treating or reducing the progression of new onset type 1 diabetes (T1D) when said antibody or fragment is administered to human.