Antigen Binding Constructs

Abstract

The present invention is directed to antigen binding constructs comprising one or two epitope binding domains separated by a single chain Fc region of an antibody, wherein each epitope binding domain in capable of binding to VEGF, to dimers comprising two antigen binding constructs of the invention, pharmaceutical compositions comprising said dimers and their use in the treatment of diseases associated with VEGF signalling, such as diabetic macular edema (DME), wet age-related macular degeneration (Wet AMD), diabetic retinopathy, retinal vein occlusion (RVO), and corneal neovascularisation, and polynucleotide sequences encoding said antigen binding constructs.

1 .- 25 . (canceled) 
     
     
         26 . An immunoglobulin single variable domain capable of binding to VEGF, wherein the immunoglobulin single variable domain comprises CDR sequences selected from the group consisting of: SEQ ID NOs: 117-119; SEQ ID NOs: 120-122; SEQ ID NOs: 123-125; SEQ ID NOs: 126-128; and SEQ ID NOs: 129-131. 
     
     
         27 . An immunoglobulin single variable domain according to  claim 26 , wherein the immunoglobulin single variable domain comprises a sequence selected from the group consisting of: SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, and SEQ ID NO: 109. 
     
     
         28 . An antigen binding construct comprising an immunoglobulin single variable domain attached to the N-terminus or the C-terminus of a single heavy chain constant region of an IgG or IgA antibody comprising one or more CH1, CH2, and CH3 constant region antibody domains, wherein the immunoglobulin single variable domain is capable of binding to VEGF and comprises CDR sequences selected from the group consisting of: SEQ ID NOs: 117-119; SEQ ID NOs: 120-122; SEQ ID NOs: 123-125; SEQ ID NOs: 126-128; and SEQ ID NOs: 129-131. 
     
     
         29 . An antigen binding construct according to  claim 28 , wherein the immunoglobulin single variable domain comprises a sequence selected from the group consisting of: SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, and SEQ ID NO: 109. 
     
     
         30 . An antigen binding construct according to  claim 29 , wherein the immunoglobulin single variable domains comprises:
 a C-terminal sequence consisting of the sequence VTVS(S)nX as shown in SEQ ID NO: 219 for a VH immunoglobulin single variable domain or VEI(K)p(R)qX as shown in SEQ ID NO: 220 for a VL immunoglobulin single variable domain; wherein:   n represents an integer independently selected from 0 or 1;   p and q each represent 0 or 1 such that when p represents 1 q may be 0 or 1 and such that when p represents 0, q also represents 0;   X may be present or absent, and if present represents an amino acid extension of 1 to 8 amino acid residues.   
     
     
         31 . An antigen binding construct according to  claim 30 , wherein for a VL immunoglobulin single variable domain the C-terminal sequence is VEI(K)p(R)qX as shown in SEQ ID NO: 220 and p is 1, q is 1 and X is A, AAA or T. 
     
     
         32 . An antigen binding construct according to  claim 28 , wherein the single heavy chain constant region comprises a sequence selected from the group consisting of SEQ ID NOs: 83, 84, and 110. 
     
     
         33 . An antigen binding construct according to  claim 28 , wherein the immunoglobulin single variable domain is attached to the single heavy chain constant region through a linker. 
     
     
         34 . An antigen binding construct according to  claim 33 , wherein the linker through which the immunoglobulin single variable domain attaches to the N-terminus of the single heavy chain constant region comprises a sequence selected from the group consisting of SEQ ID NOs: 76-82. 
     
     
         35 . An antigen binding construct according to  claim 33 , wherein the linker through which the immunoglobulin single variable domain attaches to the C-terminus of the single heavy chain constant region comprises a sequence selected from the group consisting of SEQ ID NOs: 85-94. 
     
     
         36 . An antigen binding construct according to  claim 28 , wherein the immunoglobulin single variable domain comprises the amino acid sequence shown in SEQ ID NO: 105. 
     
     
         37 . An antigen binding construct according to  claim 28 , wherein the antigen binding construct comprises a sequence selected from the group consisting of: SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174. SEQ ID NO: 175, and SEQ ID NO: 176. 
     
     
         38 . A dimer comprising two antigen binding constructs according to  claim 28 , wherein said dimer is a heterodimer or homodimer. 
     
     
         39 . An antigen binding construct comprising two immunoglobulin single variable domains separated by a single heavy chain constant region of an IgG or IgA antibody comprising one or more CH1, CH2, and CH3 constant region antibody domains, wherein each immunoglobulin single variable domain is capable of binding to VEGF and further wherein the first immunoglobulin single variable domain is attached to the N-terminus of the single heavy chain constant region and comprises CDR sequences selected from the group consisting of: SEQ ID NOs: 111-113; SEQ ID NOs: 114-116; SEQ ID NOs: 117-119; SEQ ID NOs: 120-122; SEQ ID NOs: 123-125; SEQ ID NOs: 126-128; and SEQ ID NOs: 129-131, and the second immunoglobulin single variable domain is attached to the C-terminus of the single heavy chain constant region and comprises CDR sequences selected from the group consisting of: SEQ ID NOs: 111-113; SEQ ID NOs: 114-116; SEQ ID NOs: 117-119; SEQ ID NOs: 120-122; SEQ ID NOs: 123-125; SEQ ID NOs: 126-128; and SEQ ID NOs: 129-131. 
     
     
         40 . An antigen binding construct according to  claim 39 , wherein the first immunoglobulin single variable domain comprises a sequence selected from the group consisting of SEQ ID NOs: 103-109 and wherein the second immunoglobulin single variable domain comprises a sequence selected from the group consisting of SEQ ID NOs: 103-109. 
     
     
         41 . An antigen binding construct according to  claim 39 , wherein the single heavy chain constant region comprises a sequence selected from the group consisting of SEQ ID NOs: 83, 84, and 110. 
     
     
         42 . An antigen binding construct according to  claim 39 , wherein each of the first and second immunoglobulin single variable domains is attached to the single heavy chain constant region through a linker. 
     
     
         43 . An antigen binding construct according to  claim 42 , wherein the linker through which the first immunoglobulin single variable domain attaches to the N-terminus of the single heavy chain constant region comprises a sequence selected from the group consisting of SEQ ID NOs: 76-82 and wherein the linker through which the second immunoglobulin single variable domain attaches to the C-terminus of the single heavy chain constant region comprises a sequence selected from the group consisting of SEQ ID NOs: 85-94. 
     
     
         44 . An antigen binding construct according to  claim 39 , wherein one or more immunoglobulin single variable domains comprise:
 a C-terminal sequence consisting of the sequence VTVS(S)nX as shown in SEQ ID NO: 219 for a VH immunoglobulin single variable domain or VEI(K)p(R)qX as shown in SEQ ID NO: 220 for a VL immunoglobulin single variable domain; wherein:   n represents an integer independently selected from 0 or 1;   p and q each represent 0 or 1 such that when p represents 1 q may be 0 or 1 and such that when p represents 0, q also represents 0;   X may be present or absent, and if present represents an amino acid extension of 1 to 8 amino acids residues.   
     
     
         45 . An antigen binding construct according to  claim 44 , wherein for a VL immunoglobulin single variable domain the C-terminal sequence is VEI(K)p(R)qX as shown in SEQ ID NO: 220 and p is 1, q is 1 and X is A, AAA or T. 
     
     
         46 . A dimer comprising two antigen binding constructs according to  claim 39 , wherein said dimer is a heterodimer or homodimer. 
     
     
         47 . A pharmaceutical composition comprising a dimer according to  claim 46 , and one or more pharmaceutically acceptable carrier(s). 
     
     
         48 . A method of treating ocular disease or cancer in a human in need thereof, the method comprising administering to the human an effective amount of the dimer according to  claim 46 , thereby treating ocular disease or cancer in the human. 
     
     
         49 . The method of  claim 48 , wherein the ocular disease is diabetic macular edema (DME), wet age-related macular degeneration (AMD), diabetic retinopathy, retinal vein occlusion (RVO), or corneal neovascularisation. 
     
     
         50 . A polynucleotide sequence encoding an antigen binding construct according to claim  14 . 
     
     
         51 . A recombinant transformed or transfected host cell comprising one or more polynucleotide sequences encoding an antigen binding construct according to  claim 39 . 
     
     
         52 . A method for the production of an antigen binding construct which method comprises the step of culturing a host cell comprising one or more polynucleotide sequences encoding an antigen binding construct according to  claim 39  and isolating the antigen binding construct. 
     
     
         53 . An antigen binding construct produced by the method according to  claim 52 .