Fc-region variants with modified fcrn- and protein a-binding properties

Abstract

Herein is reported a heterodimeric polypeptide comprising a first polypeptide comprising in N-terminal to C-terminal direction at least a portion of an immunoglobulin hinge region, which comprises one or more cysteine residues, an immunoglobulin CH2-domain and an immunoglobulin CH3-domain, and a second polypeptide comprising in N-terminal to C-terminal direction at least a portion of an immunoglobulin hinge region, which comprises one or more cysteine residues, an immunoglobulin CH2-domain and an immunoglobulin CH3-domain, wherein the first polypeptide comprises the mutations Y349C, T366S, L368A and Y407V (hole-chain) and the second polypeptide comprises the mutations S354C and T366W (knob-chain), and wherein the first polypeptide (hole-chain) comprises the mutations i) I253A or I253G, and ii) L314A or L314G or L314D, and wherein the first polypeptide and the second polypeptide are connected by one or more disulfide bridges, and wherein the CH3-domain of the first polypeptide and the CH3-domain of the second polypeptide both bind or both do not bind to protein A (numbering according to the Kabat EU index).

1 - 15 . (canceled) 
     
     
         16 . A method for treating an ocular vascular disease, the method comprising administering to a subject in need thereof a composition comprising an effective amount of a heterodimeric polypeptide to transport a soluble receptor ligand from the eye to the blood, wherein the heterodimeric polypeptide specifically binds the ligand and wherein the heterodimeric polypeptide comprises:
 a first polypeptide comprising in N-terminal to C-terminal direction at least a portion of an immunoglobulin hinge region, which comprises one or more cysteine residues, an immunoglobulin CH2-domain and an immunoglobulin CH3-domain, and a second polypeptide comprising in N-terminal to C-terminal direction at least a portion of an immunoglobulin hinge region, which comprises one or more cysteine residues, an immunoglobulin CH2-domain and an immunoglobulin CH3-domain,   wherein the first polypeptide comprises the mutations Y349C, T366S, L368A and Y407V (hole-chain) and the second polypeptide comprises the mutations S354C and T366W (knob-chain), and   wherein the first polypeptide (hole-chain) comprises one of the mutations selected from each of i) and ii), wherein i) and ii) are:
 (i) I253A or I253G, and 
 (ii) L314A or L314G or L314D, 
   and   wherein the first polypeptide and the second polypeptide are connected by one or more disulfide bridges, and   wherein the CH3-domain of the first polypeptide and the CH3-domain of the second polypeptide both bind or both do not bind to protein A, and   wherein the knob-chain does not comprise I253A, I253G, L314A, L314G or L314D mutations (numbering according to the Kabat EU index).   
     
     
         17 . The method according to  claim 16 , wherein the heterodimeric polypeptide further comprises:
 a) the mutation T250Q, or   b) the mutations T250Q and one of the mutations T256E and T256A, or c) one of the mutations T256E and T256A.   
     
     
         18 . The method according to  claim 16 , wherein the heterodimeric polypeptide further comprises:
 a) one of the mutations L251A, L251G, and L251D, or   b) one of the mutations H310A and H310G.   
     
     
         19 . The method according to  claim 16 , wherein the heterodimeric polypeptide further comprises at least one mutation selected from one or more of i) to vi), wherein i) to vi) are:
 (i) T250Q,   (ii) M252Y,   (iii) S254T,   (iv) T256E or T256A,   (v) T307A or T307H or T307Q or T307P, and   (vi) Q311H.   
     
     
         20 . The method according to  claim 16 , wherein the immunoglobulin hinge regions, the immunoglobulin CH2-domains, and the immunoglobulin CH3-domains of the first and second polypeptides are of the human IgG1 subclass. 
     
     
         21 . The method according to  claim 16 , wherein the first polypeptide and the second polypeptide further comprise the mutations L234A and L235A. 
     
     
         22 . The method according to  claim 16 , wherein the first polypeptide and the second polypeptide further comprise the mutation P329G. 
     
     
         23 . The method according to  claim 16 , wherein the immunoglobulin hinge regions, the immunoglobulin CH2-domains, and the immunoglobulin CH3-domains of the first and second polypeptides are of the human IgG4 subclass. 
     
     
         24 . The method according to  claim 16 , wherein the first polypeptide and the second polypeptide further comprise the mutations S228P and L235E. 
     
     
         25 . The method according to  claim 16 , wherein the first polypeptide and the second polypeptide further comprise the mutation P329G. 
     
     
         26 . The method according to  claim 16 , wherein the heterodimeric polypeptide is a full-length bispecific antibody. 
     
     
         27 . The method according to  claim 18 , wherein the heterodimeric polypeptide further comprises:
 a) the mutation T250Q, or   b) the mutation T250Q and one of the mutations T256E and T256A, or   c) one of the mutations T256E and T256A.   
     
     
         28 . The method according to  claim 18 , wherein the heterodimeric polypeptide further comprises:
 a) one of the mutations L251A, L251G, and L251D, and   b) one of the mutations H310A and H310G.   
     
     
         29 . The method according to  claim 27 , wherein the heterodimeric polypeptide further comprises:
 a) one of the mutations L251A, L251G, and L251D, and   b) one of the mutations H310A and H310G.   
     
     
         30 . The method according to  claim 27 , wherein the heterodimeric polypeptide further comprises:
 (i) a mutation selected from T307A, T307H, T307Q, and T307P, or   (ii) one or more of the mutations selected from the group consisting of Q311H, M252Y, and S254T.   
     
     
         31 . The method according to  claim 30 , wherein the heterodimeric polypeptide further comprises:
 (i) a mutation selected from T307A, T307H, T307Q, and T307P, and   (ii) one or more of the mutations selected from the group consisting of Q311H, M252Y, and S254T.   
     
     
         32 . The method according to  claim 16 , wherein the composition is administered by topical application to the cornea or by intravitreal application. 
     
     
         33 . The method according to  claim 16 , wherein the ocular vascular disease is selected from the group consisting of wet age-related macular degeneration (wet AMD), dry age-related macular degeneration (dry AMD), diabetic macular edema (DME), cystoid macular edema (CME), non-proliferative diabetic retinopathy (NPDR), proliferative diabetic retinopathy (PDR), cystoid macular edema, vasculitis (e.g. central retinal vein occlusion), papilloedema, retinitis, conjunctivitis, uveitis, choroiditis, multifocal choroiditis, ocular histoplasmosis, blepharitis, and dry eye (Sjogren's disease). 
     
     
         34 . The method according to  claim 16 , further comprising administering one or more additional therapeutic agents for a vascular eye disease. 
     
     
         35 . The method according to  claim 16 , wherein the heterodimeric polypeptide acts to inhibit angiogenesis in the eye of said subject.