Antibodies against g-csfr and uses thereof

Abstract

The present disclosure provides proteins comprising antigen binding domains of antibodies that bind to human granulocyte-colony stimulating factor receptor.

1 .- 14 . (canceled) 
     
     
         15 . A protein comprising an antigen binding site of an antibody, wherein the antigen binding site binds to human granulocyte-colony stimulating factor receptor (hG-CSFR) and neutralizes granulocyte-colony stimulating factor (G-CSF) signaling, wherein the protein comprises a V H  and a V L , and wherein:
 (i) the V H  comprises:   a complementarity determining region (CDR) 1 comprising the sequence set forth in SEQ ID NO: 6, or comprising a sequence having fewer than 5 amino acid substitutions relative to the sequence of SEQ ID NO: 6;   a CDR2 comprising the sequence set forth in SEQ ID NO: 7, or comprising a sequence having fewer than 5 amino acid substitutions relative to the sequence of SEQ ID NO: 7; and   a CDR3 comprising the sequence a sequence set forth in SEQ ID NO: 8, or comprising a sequence having fewer than 6 amino acid substitutions relative to the sequence of SEQ ID NO: 8; and/or   (ii) the V L  comprises:   a CDR1 comprising the sequence set forth in SEQ ID NO: 9, or comprising a sequence having fewer than 5 amino acid substitutions relative to the sequence of SEQ ID NO: 9;   a CDR2 comprising the sequence set forth in SEQ ID NO: 10, or comprising a sequence having fewer than 5 amino acid substitutions relative to the sequence of SEQ ID NO: 10 and   a CDR3 comprising the sequence as set forth in SEQ ID NO: 11, or comprising a sequence having fewer than 6 amino acid substitutions relative to the sequence of SEQ ID NO:  11 .   
     
     
         16 . The protein of  claim 15 , wherein:
 (i) the V H  comprises:   
       a complementarity determining region (CDR) 1 comprising the sequence set forth in SEQ ID NO: 6;
 a CDR2 comprising the sequence set forth in SEQ ID NO: 7; and 
 a CDR3 comprising the sequence a sequence set forth in SEQ ID NO: 8, or comprising a sequence having fewer than 6 amino acid substitutions relative to the sequence of SEQ ID NO: 8; and/or 
 (ii) the V L  comprises: 
 a CDR1 comprising the sequence set forth in SEQ ID NO: 9; 
 a CDR2 comprising the sequence set forth in SEQ ID NO: 10; and 
 a CDR3 comprising the sequence as set forth in SEQ ID NO: 11, or comprising a sequence having fewer than 6 amino acid substitutions relative to the sequence of SEQ ID NO:  11 . 
 
     
     
         17 . The protein of  claim 15 , wherein:
 (i) the V H  comprises:   
       a complementarity determining region (CDR) 1 comprising the sequence set forth in SEQ ID NO: 6; a CDR2 comprising the sequence set forth in SEQ ID NO: 7; and a CDR3 comprising the sequence set forth in SEQ ID NO: 8, or a CDR3 comprising a sequence having fewer than 5 amino acid substitutions relative to the sequence of SEQ ID NO: 8; or
 a sequence set forth in SEQ ID NO: 2, SEQ ID NO: 4, or a sequence having fewer than 5 amino acid substitutions relative to the sequence of SEQ ID NO: 2 or SEQ ID NO: 4; and/or 
 (ii) the V L  comprises: 
 a CDR1 comprising the sequence set forth in SEQ ID NO: 9; a CDR2 comprising the sequence set forth in SEQ ID NO: 10; and a CDR3 comprising the sequence set forth in SEQ ID NO: 11 or a CDR3 comprising a sequence having fewer than 4 amino acid substitutions relative to the sequence of SEQ ID NO: 11; or 
 a sequence set forth in SEQ ID NO: 3, SEQ ID NO: 5, or a sequence having fewer than 5 amino acid substitutions relative to the sequence of SEQ ID NO: 3 or SEQ ID NO: 5. 
 
     
     
         18 . The protein of  claim 15 , wherein:
 (i) the V H  comprises:   a complementarity determining region (CDR) 1 comprising the sequence set forth in SEQ ID NO: 6; a CDR2 comprising the sequence set forth in SEQ ID NO: 7; and a CDR3 comprising the sequence set forth in SEQ ID NO: 8, or a CDR3 comprising a sequence having fewer than 5 amino acid substitutions relative to the sequence of SEQ ID NO: 8; or   a sequence set forth in SEQ ID NO: 2, SEQ ID NO: 4, or a sequence having fewer than 5 amino acid substitutions relative to the sequence of SEQ ID NO: 2 or SEQ ID NO: 4; and   (ii) the V L  comprises:   a CDR1 comprising the sequence set forth in SEQ ID NO: 9; a CDR2 comprising the sequence set forth in SEQ ID NO: 10; and a CDR3 comprising the sequence set forth in SEQ ID NO: 11 or a CDR3 comprising a sequence having fewer than 4 amino acid substitutions relative to the sequence of SEQ ID NO: 11; or   a sequence set forth in SEQ ID NO: 3, SEQ ID NO: 5, or a sequence having fewer than 5 amino acid substitutions relative to the sequence of SEQ ID NO: 3 or SEQ ID NO: 5.   
     
     
         19 . The protein of  claim 15 , wherein the protein competitively inhibits the binding of monoclonal antibody C1.2 or monoclonal antibody C1.2G to one or more of:
 (i) a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the lysine at position 167 of SEQ ID NO: 1; and/or   (ii) a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the histidine at position 168 of SEQ ID NO: 1,   wherein C1.2 comprises a V H  comprising a sequence set forth in SEQ ID NO: 2 and a V L  comprising a sequence set forth in SEQ ID NO: 3, and C1.2G comprises a V H  comprising a sequence set forth in SEQ ID NO: 4 and a V L  comprising a sequence set forth in SEQ ID NO: 5,   
     
     
         20 . The protein of  claim 15 , wherein the antigen binding site also binds to the polypeptide at (i) and/or (ii), and wherein the level of binding of the protein to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for any one of:
 (a) the arginine at position 287 of SEQ ID NO:1;   (b) the histidine at position 237 of SEQ ID NO:1;   (c) the methionine at position 198 of SEQ ID NO:1;   (d) the tyrosine at position 172 of SEQ ID NO:1;   (e) the leucine at position 171 of SEQ ID NO:1; or   (f) the leucine at position 111 of SEQ ID NO:1   is lower than the level of binding of the protein to a polypeptide of SEQ ID NO: 1.   
     
     
         21 . The protein of  claim 15 , wherein the antigen binding domain binds to an epitope comprising residues within at least one region selected from amino acids 111-115, 170-176, 218-234 and/or 286-300 of SEQ ID NO: 1. 
     
     
         22 . The protein of  claim 15 , wherein upon binding of the protein to a polypeptide of SEQ ID NO: 1 and cleavage using proteolytic enzymes, the protein remains bound to at least one peptide comprising amino acids 111-115 of SEQ ID NO: 1, amino acids 170-176 of SEQ ID NO: 1, amino acids 218-234 of SEQ ID NO: 1, or amino acids 286-300 of SEQ ID NO: 1. 
     
     
         23 . The protein of  claim 15  comprising:
 (i) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 2 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 3; 
 (ii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 5; 
 (iii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 15 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 14; 
 (iv) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 16; 
 (v) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 17; 
 (vi) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 18; 
 (vii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 20 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 19; 
 (viii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 21; 
 (ix) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 22; 
 (x) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 24 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 23; 
 (ix) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 25; 
 (x) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 26; 
 (xi) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 28 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 27; 
 (xii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 29; 
 (xiii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 31 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 30; 
 (xiv) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 33 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 32; 
 (xv) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 34; 
 (xvi) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 36 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 35; 
 (xvii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 38 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 37; 
 (xviii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 40 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 39; 
 (xix) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 42 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 41; 
 (xx) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 43; 
 (xxi) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 45 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 44; 
 (xxii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 46; 
 (xxiii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 47; 
 (xxiv) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 49 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 48; 
 (xxv) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 51 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 50; 
 (xxvi) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 53 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 52; 
 (xxvii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 54; 
 (xxviii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 55 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 5; 
 (xxix) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 57 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 56; 
 (xxx) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 59 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 58; 
 (xxxi) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 61 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 60; 
 (xxxii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 62; 
 (xxxiii) a V H  comprising an amino acid sequence set forth in SEQ ID NO: 4 and a V L  comprising an amino acid sequence set forth in SEQ ID NO: 63; and 
 (xxxix) a V H  comprising three CDRs of a V H  set forth in any one or more of (i) to (xxxiii) and a V L  comprising three CDRs of a V L  set forth in any one or more of (i) to (xxxiii). 
 
     
     
         24 . The protein of  claim 15  conjugated to another compound. 
     
     
         25 . A composition comprising the protein of  claim 15  and a pharmaceutically acceptable carrier. 
     
     
         26 . A method for treating a granulocyte-colony stimulating factor (G-CSF)-mediated condition in a subject, the method comprising administering the protein of  claim 15  to the subject. 
     
     
         27 . The method of  claim 26  comprising administering an amount of the protein sufficient to reduce the number of neutrophils in a subject without inducing neutropenia. 
     
     
         28 . The method of  claim 27 , wherein the amount of the protein is between 0.05 mg/kg and 30 mg/kg.