Fc-receptor binding modified asymmetric antibodies and methods of use

Abstract

Herein is reported an IgG class Fc-region comprising a first variant Fc-region polypeptide and a second variant Fc-region polypeptide, wherein the first variant Fc-region polypeptide is derived from a first parent IgG class Fc-region polypeptide and the second variant Fc-region polypeptide is derived from a second parent IgG class Fc-region polypeptide, whereby the first parent IgG class Fc-region polypeptide is identical to or different from the second parent IgG class Fc-region polypeptide, and the first variant Fc-region polypeptide differs from the second variant Fc-region polypeptide in one or more amino acid residues other than those amino acid residues in which the first parent IgG class Fc-region polypeptide differs from the second parent IgG class Fc-region polypeptide, and the IgG class Fc-region comprising the first variant Fc-region polypeptide and the second variant Fc-region peptide has an affinity to a human Fc-receptor that is different than that of an IgG class Fc-region comprising the first parent IgG class Fc-region polypeptide of a) and the second parent IgG class Fc-region polypeptide of a).

1 - 43 . (canceled) 
     
     
         44 . An antibody, comprising a first Fc-region polypeptide and a second Fc-region polypeptide,
 wherein the first and a second Fc-region polypeptide are both of human IgG1 or human IgG4 subclass and comprise one of the mutations selected from the group H310A, H433A and Y436A in the first Fc-region polypeptide and two of the mutations selected from the group comprising the mutations H310A, H433A and Y436A in the second Fc-region polypeptide so that all of the mutations in the first and the second Fc-region polypeptide when taken together result in that the mutations H310A, H433A and Y436A are comprised in the Fc-region,   wherein the amino acid at position 2.51 is L in the first and the second Fc-region polypeptide,   wherein the amino acid at position 253 is I in the first and the second Fc-region polypeptide, and   wherein the amino acid at position 435 is H in the first and the second Fc-region polypeptide   (numbering according to Kabat EU index numbering system).   
     
     
         45 . The antibody according to  claim 44 , wherein the antibody is a monoclonal antibody. 
     
     
         46 . The antibody according to  claim 44 , wherein the antibody is a human, humanized or chimeric antibody. 
     
     
         47 . The antibody according to  claim 44 , wherein the antibody is a bispecific antibody. 
     
     
         48 . The antibody according to  claim 44 , wherein the antibody is a bivalent antibody. 
     
     
         49 . The antibody according to  claim 44 , wherein the antibody has the same binding to  Staphylococcus  protein A as an antibody with a wild-type human IgG1 or IgG4 class Fc-region. 
     
     
         50 . The antibody according to  claim 44 , wherein:
 i) the first Fc-region polypeptide is selected from the group consisting of
 human IgG1 Fc-region polypeptide, 
 human IgG4 Fc-region polypeptide, 
 human IgG1 Fc-region polypeptide with the mutations L234A, L235A, 
 human IgG1 Fc-region polypeptide with the mutations Y349C, T366S, L368A, Y407V, 
 human IgG1 Fc-region polypeptide with the mutations S354C, T366S, L368A, Y407V, 
 human IgG1 Fc-region polypeptide with the mutations L234A, L235A, Y349C, T366S, L368A, Y407V, 
 human IgG1, Fc-region polypeptide with the mutations L234A, L235A, S354C, T366S, L368A, Y407V, 
 human IgG1 Fc-region polypeptide with the mutations P329G, 
 human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G, 
 human IgG1 Fc-region polypeptide with the mutations P329G, Y349C, T366S, L368A, Y407V, 
 human IgG1 Fc-region polypeptide with the mutations P329G, S354C, T366S, L368A, Y407V, 
 human IgG Fc-region polypeptide with the mutations L234A, L235A, P329G, Y349C, T366S, L368A, Y407V, 
 human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G, S354C, T366S, L368A, Y40717, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, P329G, 
 human IgG4 Fc-region polypeptide with the mutations Y349C, T366S, L368A, Y407V, 
 human IgG4 Fc-region polypeptide with the mutations S354C, T366S, L368A, Y407V, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, Y349C, T366S, L368A, Y407V, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, S354C, T366S, L368A, Y407V, 
 human IgG4 Fc-region polypeptide with the mutations P329G, 
 human IgG4 Fc-region polypeptide with the mutations P329G, Y349C, T366S, L368A, Y407V, 
 human IgG4 Fc-region polypeptide with the mutations P329G, S354C, T366S, L368A, Y407V, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, P329G, Y349C, T366S, L368A, Y407V, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, P329G, S354C, T366S, L368A, Y407V, and 
 human IgG1 or IgG4⋅with the mutations K392D, 
   and   ii) the second Fc-region polypeptide is selected from the group consisting of
 human IgG1 Fc-region polypeptide, 
 human IgG4 Fc-region polypeptide, 
 human IgG1 Fc-region polypeptide with the mutations L234A, L235A, 
 human IgG1 Fc-region polypeptide with the mutations S354C, T366W, 
 human IgG1 Fc-region polypeptide with the mutations Y349C, T366W, 
 human IgG Fc-region polypeptide with the mutations L234A, L235A, S354C, T366W, 
 human IgG1 Fc-region polypeptide with the Mutations L234A, L235A, Y349C, T366W, 
 human IgG1 Fc-region polypeptide with the mutations P329G, 
 human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G, 
 human IgG Fc-region polypeptide with the mutations P329G, S354C, T366W, 
 human IgG1 Fc-region polypeptide with the mutations P329G, Y349C, T366W, 
 human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G, S354C, T366W, 
 human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G, Y349C, T366W, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, P329G, 
 human IgG4 Fc-region polypeptide with the mutations S354C, T366W, 
 human IgG4 Fc-region polypeptide with the mutations Y349C, T366W, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, S354C, T366W, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, Y349C, T366W, 
 human IgG4 Fc-region polypeptide with the mutations P329G, 
 human IgG4 Fc-region polypeptide with the mutations P329G, S354C, T366W, 
 human IgG4 Fc-region polypeptide with the mutations P329G, Y349C, T366W, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, P329G, S354C, T366W, 
 human IgG4 Fc-region polypeptide with the mutations S228P, L235E, P329G, Y349C, T366W, 
 human IgG1 with the mutations D399K, D356K, and/or E357K, and 
 human IgG4 with the mutations D399K, E356K, and/or E357K. 
   
     
     
         51 . The antibody according to  claim 44 , wherein:
 i) the first Fc-region polypeptide is a human IgG1 Fc-region polypeptide and the second Fc-region polypeptide is a human IgG1 Fc-region polypeptide, or   ii) the first Fc-region polypeptide is a human IgG Fc-region polypeptide with the mutations L234A, L235A and the second Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, or   iii) the first Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G and the second Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G, or   iv) the first Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, S354C, T366W and the second Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, Y349C, T366S, L368A, Y407V, or   v) the first Fc-region polypeptide is a human IgG-1 Fc-region polypeptide with the mutations L234A, L235A, P329G, S354C, T366W and the second Fc-region polypeptide is a human IgG1 Fc-region polypeptide with the mutations L234A, L235A, P329G, Y349C, T366S, L368A, Y407V, or   vi) the first Fc-region polypeptide is a human IgG4 Fc-region polypeptide and the second Fc-region polypeptide is a human IgG4 Fc-region polypeptide, or   vii) the first Fc-region polypeptide is a human IgG4 Fc-region polypeptide with the mutations S228P, L235E and the second Fc-region polypeptide is a human IgG4 Fc-region polypeptide with the mutations S228P, L235E, or   viii) the first Fc-region polypeptide is a human IgG4 Fc-region polypeptide with the mutations S228P, L235E, P329G and the second Fc-region polypeptide is a human IgG4 Fc-region polypeptide with the mutations S228P, L235E, P329G, or   ix) the first Fc-region polypeptide is a human IgG4 Fc-region polypeptide with the mutations S228P, L235E, S354C, T366W and the second Fc-region polypeptide is a human IgG4 Fc-region polypeptide with the mutations S228P, L235E, Y349C, T366S, L368A, Y407V, or   the first Fc-region polypeptide is a human IgG4 Fc-region polypeptide with the mutations S228P, L235E, P329G, S354C, T366W and the second Fc-region polypeptide is a human IgG4 Fc-region polypeptide with the mutations S228P, L235E, P329G, Y349C, T366S, L368A, Y407V.   
     
     
         52 . A nucleic acid encoding the antibody according to  claim 44 . 
     
     
         53 . A cell comprising the nucleic acid according to  claim 52 . 
     
     
         54 . A method of producing the antibody according to  claim 44  comprising the steps of
 culturing the host cell of  claim 53 , and 
 recovering the antibody from the cell or the cultivation medium. 
 
     
     
         55 . A pharmaceutical formulation, comprising the antibody according to  claim 44 . 
     
     
         56 . A method for the treatment of an ocular vascular disease, comprising the step of administering the antibody according to  claim 44  to a patient in need thereof.