THERAPEUTIC ANTIGEN-BINDING MOLECULE WITH A FcRn-BINDING DOMAIN THAT PROMOTES ANTIGEN CLEARANCE

Abstract

The present invention provides: a modified FcRn-binding domain having an enhanced affinity for the Fc Receptor neonatal (FcRn) at neutral pH; an antigen-binding molecule comprising said FcRn-binding domain, which has low immunogenicity, high stability and form only a few aggregates; a modified antigen-binding molecule having an increased FcRn-binding activity at neutral or acidic pH without an increased binding activity at neutral pH for a pre-existing anti-drug antibody; use of the antigen-binding molecules for improving antigen-binding molecule-mediated antigen uptake into cells; use of the antigen-binding molecules for reducing the plasma concentration of a specific antigen; use of the modified FcRn-binding domain for increasing the total number of antigens to which a single antigen-binding molecule can bind before its degradation; use of the modified FcRn-binding domain for improving pharmacokinetics of an antigen-binding molecule; methods for decreasing the binding activity for a pre-existing anti-drug antibody; and methods for producing said antigen-binding molecules.

1 - 61 . (canceled) 
     
     
         62 . An antigen-binding molecule comprising a modified FcRn-binding domain, the amino acid sequence of which differs from the sequence of a native human IgG FcRn-binding domain at one or more positions, including an amino acid substitution at at least one of the following positions (EU numbering): 238, 250, 252, 254, 255, 258, 286, 307, 308, 309, 311, 315, 428, 433, 434, and 436. 
     
     
         63 . The antigen-binding molecule of  claim 62 , wherein the at least one position that is substituted includes positions 252 and 434 and one or more of the following positions: 238, 250, 254, 255, 256, 258, 286, 387, 307, 308, 309, 311, 315, 428, 433, and 436 (all positions by EU numbering). 
     
     
         64 . The antigen-binding molecule of  claim 62 , wherein at least one of the following positions in the modified FcRn-binding domain is occupied by the indicated amino acid (all positions by EU numbering):
 aspartic acid at position 238;   valine at position 250;   tyrosine at position 252;   threonine at position 254;   leucine at position 255;   glutamic acid at position 256;   either aspartic acid or isoleucine at position 258;   glutamic acid at position 286;   glutamine at position 307;   proline at position 308;   glutamic acid at position 309;   either alanine or histidine at position 311;   aspartic acid at position 315;   isoleucine at position 428;   any one of alanine, lysine, proline, arginine, and serine at position 433;   either tyrosine or tryptophan at position 434;   any one of isoleucine, leucine, valine, threonine, and phenylalanine at position 436.   
     
     
         65 . The antigen-binding molecule of  claim 62 , wherein the at least one position that is substituted includes all members of a set of positions selected from any of sets (a)-(g):
 (a) positions 252, 434, and 436;   (b) positions 252, 307, 311 and 434;   (c) positions 252, 315, and 434;   (d) positions 252, 308, and 434;   (e) positions 238, 252, and 434;   (f) positions 252, 434, 307, 311, and 436;   (g) positions 252, 387, and 434 (all positions by EU numbering).   
     
     
         66 . The antigen-binding molecule of  claim 62 , wherein the modified FcRn-binding domain comprises one of the following combinations of amino acid residues at the indicated positions:
 (a) tyrosine at position 252, aspartic acid at position 315, and tyrosine at position 434;   (b) tyrosine at position 252, tyrosine at position 434, and isoleucine at position 436;   (c) tyrosine at position 252, tyrosine at position 434, and leucine at position 436;   (d) tyrosine at position 252, tyrosine at position 434, and valine at position 436; or   (e) tyrosine at position 252, threonine at position 254, tyrosine at position 434, and isoleucine at position 436 (all positions by EU numbering).   
     
     
         67 . The antigen-binding molecule of  claim 62 , wherein the at least one position that is substituted includes all members of a set of positions selected from any of sets (a)-(h):
 (a) positions 252, 434, 307, 311, and 286;   (b) positions 252, 434, 307, 311, 286, and 254;   (c) positions 252, 434, 307, 311, and 436;   (d) positions 252, 434, 307, 311, 436, and 254;   (e) positions 252, 434, 307, 311, 436, and 250;   (f) positions 252, 434, 308, and 250;   (g) positions 252, 434, 308, 250, and 436;   (h) positions 252, 434, 308, 250, 307, and 311 (all positions by EU numbering).   
     
     
         68 . The antigen-binding molecule of  claim 62 , wherein the modified FcRn-binding domain comprises one of the following combinations of amino acid residues at the indicated positions:
 (a) tyrosine at position 252, glutamic acid at position 286, glutamine at position 307, alanine at position 311, and tyrosine at position 434;   (b) tyrosine at position 252, threonine at position 254, glutamic acid at position 286, glutamine at position 307, alanine at position 311, and tyrosine at position 434;   (c) tyrosine at position 252, glutamine at position 307, alanine at position 311, tyrosine at position 434, and isoleucine at position 436;   (d) tyrosine at position 252, threonine at position 254, glutamic acid at position 286, glutamine at position 307, alanine at position 311, tyrosine at position 434, and isoleucine at position 436;   (e) valine at position 250, tyrosine at position 252, threonine at position 254, proline at position 308, tyrosine at position 434, and valine at position 436;   (f) valine at position 250, tyrosine at position 252, glutamine at position 307, alanine at position 311, tyrosine at position 434, and valine at position 436;   (g) tyrosine at position 252, glutamine at position 307, alanine at position 311, tyrosine at position 434, and valine at position 436;   (h) valine at position 250, tyrosine at position 252, proline at position 308, and tyrosine at position 434; or   (i) valine at position 250, tyrosine at position 252, glutamine at position 307, proline at position 308, alanine at position 311, and tyrosine at position 434 (all positions by EU numbering).   
     
     
         69 . The antigen-binding molecule of  claim 62 , wherein the at least one position that is substituted includes all members of a set of positions selected from any of sets (a)-(d):
 (a) positions 252, 434, 307, 311, 436, and 286;   (b) positions 252, 434, 307, 311, 436, 250, and 308;   (c) positions 252, 434, 307, 311, 436, 250, 286, and 308;   (d) positions 252, 434, 307, 311, 436, 250, 286, 308, and 428 (all positions by EU numbering).   
     
     
         70 . The antigen-binding molecule of  claim 62 , wherein the modified FcRn-binding domain comprises one of the following combinations of amino acid residues at the indicated positions:
 (a) tyrosine at position 252, glutamic acid at position 286, glutamine at position 307, alanine at position 311, tyrosine at position 434, and valine at position 436;   (b) valine at position 250, tyrosine at position 252, glutamine at position 307, proline at position 308, alanine at position 311, tyrosine at position 434, and valine at position 436;   (c) valine at position 250, tyrosine at position 252, glutamic acid at position 286, glutamine at position 307, proline at position 308, alanine at position 311, tyrosine at position 434, and valine at position 436; or   (d) valine at position 250, tyrosine at position 252, glutamic acid at position 286, glutamine at position 307, proline at position 308, alanine at position 311, tyrosine at position 434, and valine at position 436 (all positions by EU numbering).   
     
     
         71 . The antigen-binding molecule of  claim 62 , wherein the at least one position that is substituted includes all members of a set of positions selected from any of sets (a)-(c):
 (a) positions 434, 307, and 311;   (b) positions 434, 307, 309, and 311;   (c) positions 434, 250, 252, and 436 (all positions by EU numbering).   
     
     
         72 . The antigen-binding molecule of  claim 62 , wherein the modified FcRn-binding domain comprises one of the following combinations of amino acid residues at the indicated positions:
 (a) glutamine at position 307, histidine at position 311, and tyrosine at position 434;   (b) glutamine at position 307, glutamic acid at position 309, alanine at position 311, and tyrosine at position 434;   (c) glutamine at position 307, glutamic acid at position 309, histidine at position 311, and tyrosine at position 434; or   (d) valine at position 250, tyrosine at position 252, tyrosine at position 434, and valine at position 436 (all positions by EU numbering).   
     
     
         73 . The antigen-binding molecule of  claim 62 , wherein the antigen-binding molecule comprises an antigen-binding domain whose antigen-binding activity is lower at pH 5.8 than at pH 7.4. 
     
     
         74 . The antigen-binding molecule of  claim 62 , wherein the antigen-binding molecule comprises an antigen-binding domain whose antigen-binding activity is lower at a first calcium concentration that is between 0.1 μM to 30 μM than at a second calcium concentration that is between 100 μM to 10 mM. 
     
     
         75 . The antigen-binding molecule of  claim 62 , wherein the antigen-binding molecule has an FcRn receptor-binding affinity of 50-150 nM at pH 7.0, a melting temperature (Tm) above 63.0° C., and an Epibase score lower than 250. 
     
     
         76 . The antigen-binding molecule of  claim 62 , wherein the antigen-binding molecule has an FcRn receptor binding affinity of 15-50 nM at pH 7.0, a Tm above 60° C., and an Epibase score lower than 500. 
     
     
         77 . The antigen-binding molecule of  claim 62 , wherein the antigen-binding molecule has an FcRn receptor binding affinity stronger than 15 nM at pH 7.0, a Tm higher than 57.5° C., and an Epibase score lower than 500. 
     
     
         78 . The antigen-binding molecule of  claim 62 , wherein the at least one position that is substituted includes (a) one or more of positions 238, 255, and 258; and (b) three or more additional positions, wherein the three or more additional positions are a combination of positions selected from any of the combinations listed in any of Tables 4 to 7 (all positions by EU numbering). 
     
     
         79 . The antigen-binding molecule of  claim 62 , wherein the amino acid at position 257 (EU numbering) of the modified FcRn-binding domain is not alanine, valine, isoleucine, leucine, or threonine. 
     
     
         80 . The antigen-binding molecule of  claim 62 , wherein the amino acid at position 252 (EU numbering) of the modified FcRn-binding domain is not tryptophan. 
     
     
         81 . The antigen-binding molecule of  claim 62 , wherein a rheumatoid factor (RF) antibody's binding affinity for the modified FcRn-binding domain is the same or decreased compared to the binding affinity of the RF antibody for a native human IgG FcRn-binding domain of a control IgG antibody. 
     
     
         82 . The antigen-binding molecule of  claim 81 , wherein the difference between the sequence of the modified FcRn-binding domain and the sequence of the native human IgG FcRn-binding domain also includes a substitution at one or more of the following positions (by EU numbering): 387, 422, 424, 426, 433, 436, 438 and 440. 
     
     
         83 . The antigen-binding molecule of  claim 82 , wherein one or more of the following positions (EU numbering) in the modified FcRn-binding domain is occupied by the indicated amino acid:
 arginine at position 387,   any one of glutamic acid, arginine, serine, aspartic acid, lysine, threonine and glutamine at position 422;   any one of glutamic acid, arginine, lysine, and asparagine at position 424;   any one of aspartic acid, glutamine, alanine, and tyrosine at position 426;   aspartic acid at position 433;   threonine at position 436;   any one of glutamic acid, arginine, serine, and lysine at position 438; and   any one of glutamic acid, aspartic acid and glutamine at position 440.   
     
     
         84 . The antigen-binding molecule of  claim 62 , wherein the modified FcRn-binding domain differs from the native human IgG FcRn-binding domain by substitutions comprising a combination of substitutions selected from the combinations of three or more substitutions listed in Tables 12 and 13. 
     
     
         85 . The antigen-binding molecule of  claim 62 , wherein the modified FcRn-binding domain differs from the native human IgG FcRn-binding domain by substitutions comprising a combination of substitutions selected from the combinations listed in Tables 14 and 15. 
     
     
         86 . The antigen-binding molecule of  claim 82 , wherein the modified FcRn-binding domain comprises any one of the following sets of amino acids at the indicated positions (all positions by EU numbering):
 (a) tyrosine at position 252, arginine at position 387, tyrosine at position 434, and valine at position 436; or   (b) tyrosine at position 252, glutamic acid at position 422, tyrosine at position 434, and valine at position 436; or   (c) tyrosine at position 252, arginine at position 422, tyrosine at position 434, and valine at position 436; or   (d) tyrosine at position 252, serine at position 422, tyrosine at position 434, and valine at position 436; or   (e) tyrosine at position 252, glutamic acid at position 424, tyrosine at position 434, and valine at position 436; or   (f) tyrosine at position 252, arginine at position 424, tyrosine at position 434, and valine at position 436; or   (g) tyrosine at position 252, tyrosine at position 434, valine at position 436, and glutamic acid at position 438; or   (h) tyrosine at position 252, tyrosine at position 434, valine at position 436, and arginine at position 438; or   (i) tyrosine at position 252, tyrosine at position 434, valine at position 436, and serine at position 438; or   (j) tyrosine at position 252, tyrosine at position 434, valine at position 436, and glutamic acid at position 440.   
     
     
         87 . The antigen-binding molecule of  claim 62 , wherein the antigen-binding molecule is an antibody. 
     
     
         88 . A composition comprising the antigen-binding molecule of  claim 87 , wherein at least 98% of the antigen-binding molecule in the composition is in the form of antibody monomers containing two heavy chains and two light chains. 
     
     
         89 . A method for reducing plasma concentration of an antigen in a subject, the method comprising administering to the subject the antigen-binding molecule of  claim 62 , wherein the antigen-binding molecule binds to the antigen.