DUAL SPECIFIC BINDING PROTEINS DIRECTED AGAINST IL-1 and/or IL-17

Abstract

Engineered multivalent and multispecific binding proteins that bind IL-1β and/or IL-17 are provided, along with methods of making and uses in the prevention, diagnosis, and/or treatment of disease.

1 - 34 . (canceled) 
     
     
         35 . An isolated nucleic acid or group of nucleic acids encoding a binding protein, wherein the binding protein comprises first and second polypeptide chains, each independently comprising VD1-(X1)n-VD2-C-X2, wherein
 VD1 is a first variable domain;   VD2 is a second variable domain;   C is a constant domain;   X1 is a linker;   X2 is an Fc region that is either present or absent; and   n is 0 or 1;   
       wherein the VD1 domains on the first and second polypeptide chains form a first functional target binding site and the VD2 domains on the first and second polypeptide chains form a second functional target binding site, and wherein the binding protein is capable of binding IL-1β and IL-17, wherein
 (i) the variable domains that form a functional target binding site for IL-1β comprise:
 CDRs 1-3 from SEQ ID NO: 32 and CDRs 1-3 from SEQ ID NO: 33, 
 CDRs 1-3 from SEQ ID NO: 34 and CDRs 1-3 from SEQ ID NO: 35, 
 CDRs 1-3 from SEQ ID NO: 36 and CDRs 1-3 from SEQ ID NO: 37, 
 CDRs 1-3 from SEQ ID NO: 38 and CDRs 1-3 from SEQ ID NO: 39, or 
 CDRs 1-3 from SEQ ID NO: 40 and CDRs 1-3 from SEQ ID NO: 41; 
 
 and 
 (ii) the variable domains that form a functional target binding site for IL-17 comprise CDRs 1-3 from SEQ ID NO: 44 and CDRs 1-3 from SEQ ID NO: 45. 
 
     
     
         36 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein has a first polypeptide chain comprising SEQ ID NO: 104 and a second polypeptide chain comprising SEQ ID NO: 105. 
     
     
         37 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein comprises a first polypeptide chain comprising VD1-(X1)n-VD2-C-X2, wherein
 VD1 is a first heavy chain variable domain;   VD2 is a second heavy chain variable domain;   C is a heavy chain constant domain;   X1 is a linker;   X2 is an Fc region that is either present or absent;   n is 0 or 1, and   
       a second polypeptide chain comprising VD1-(X1)n-VD2-C, wherein
 VD1 is a first light chain variable domain; 
 VD2 is a second light chain variable domain; 
 C is a light chain constant domain; 
 X1 is a linker; 
 n is 0 or 1, 
 
       wherein the VD1 domains on the first and second polypeptide chains form a first functional target binding site and the VD2 domains on the first and second polypeptide chains form a second functional target binding site. 
     
     
         38 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein
 (i) the binding protein is capable of binding IL-1β with a K D  of about 5.1×10 −11  M, as measured by surface plasmon resonance, or capable of inhibiting IL-1β with an IC50 of about 2.563 nM, as measured in an IL-1β neutralization assay, and/or   (ii) the binding protein is capable of binding IL-17 with a K D  of about 4.8×10 −12  M, as measured by surface plasmon resonance, or capable of inhibiting IL-17 with an IC50 of about 1.7 nM, as measured in an IL-17 neutralization assay.   
     
     
         39 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein comprises
 (i) variable domains that form a functional target binding site for IL-1β comprising:
 (1) SEQ ID NO: 32 and SEQ ID NO: 33, 
 (2) SEQ ID NO: 34 and SEQ ID NO: 35, 
 (3) SEQ ID NO: 36 and SEQ ID NO: 37, 
 (4) SEQ ID NO: 38 and SEQ ID NO: 39, or 
 (5) SEQ ID NO: 40 and SEQ ID NO: 41; 
   and   (ii) variable domains that form a functional target binding site for IL-17 comprising:
 SEQ ID NO: 44 and SEQ ID NO: 45. 
   
     
     
         40 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein comprises two first polypeptide chains and two second polypeptide chains and four functional target binding sites. 
     
     
         41 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein X1 is any one of SEQ ID NOs: 1-31. 
     
     
         42 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the Fc region of the binding protein is a variant sequence Fc region. 
     
     
         43 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the Fc region of the binding protein is an Fc region from an IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, or IgD. 
     
     
         44 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein comprises
 (a) a heavy chain constant region comprising a wild type human IgG1 heavy chain sequence; and   (b) a light chain constant region comprising a wild type human kappa light chain constant region sequence.   
     
     
         45 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein comprises
 (a) a heavy chain constant region comprising a human IgG1 heavy chain sequence modified by one or more amino acid changes, wherein the changes comprise substitution of leucines at positions 234 and 235 with alanines, wherein the amino acid positions are numbered using EU index numbering; and   (b) a light chain constant region comprising a wild type human kappa light chain constant region sequence.   
     
     
         46 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein comprises
 (i) variable domains that form a functional target binding site for IL-1β comprising CDRs 1-3 from SEQ ID NO: 32 and CDRs 1-3 from SEQ ID NO: 33, and   (ii) variable domains that form a functional target binding site for IL-17 comprising CDRs 1-3 from SEQ ID NO: 44 and CDRs 1-3 from SEQ ID NO: 45.   
     
     
         47 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein comprises
 (i) variable domains that form a functional target binding site for IL-1β comprising SEQ ID NO: 32 and SEQ ID NO: 33, and   (ii) variable domains that form a functional target binding site for IL-17 comprising SEQ ID NO: 44 and SEQ ID NO: 45.   
     
     
         48 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the first polypeptide chain of the binding protein comprises SEQ ID NO: 98 and the second polypeptide chain of the binding protein comprises SEQ ID NO: 99. 
     
     
         49 . The isolated nucleic acid or group of nucleic acids of  claim 48 , wherein the binding protein comprises
 (a) a heavy chain constant region on the first polypeptide chain comprising a human IgG1 heavy chain sequence modified by one or more amino acid changes, wherein the changes comprise substitution of leucines at positions 234 and 235 with alanines, wherein the amino acid positions are numbered using EU index numbering; and   (b) a light chain constant region on the second polypeptide chain comprising a wild type human kappa light chain constant region sequence.   
     
     
         50 . The isolated nucleic acid or group of nucleic acids of  claim 48 , wherein:
 (i) the binding protein is capable of binding IL-1β with a K D  of about 5.1×10 −11  M, as measured by surface plasmon resonance, or capable of inhibiting IL-1β with an IC50 of about 0.027 nM, as measured in an IL-1β neutralization assay, and/or   (ii) the binding protein is capable of binding IL-17 with a K D  of about 4.8×10 −12  M, as measured by surface plasmon resonance, or capable of inhibiting IL-17 with an IC50 of about 0.091 nM, as measured in an IL-17 neutralization assay.   
     
     
         51 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein comprises
 (i) variable domains that form a functional target binding site for IL-1β comprising CDRs 1-3 from SEQ ID NO: 34 and CDRs 1-3 from SEQ ID NO: 35, and   (ii) variable domains that form a functional target binding site for IL-17 comprising CDRs 1-3 from SEQ ID NO: 44 and CDRs 1-3 from SEQ ID NO: 45.   
     
     
         52 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein comprises
 (i) variable domains that form a functional target binding site for IL-1β comprising SEQ ID NO: 34 and SEQ ID NO: 35, and   (ii) variable domains that form a functional target binding site for IL-17 comprising SEQ ID NO: 44 and SEQ ID NO: 45.   
     
     
         53 . The isolated nucleic acid or group of nucleic acids of  claim 51 , wherein X1 on the first polypeptide chain comprises SEQ ID NO: 29 and X1 on the second polypeptide chain comprises SEQ ID NO: 30. 
     
     
         54 . The isolated nucleic acid or group of nucleic acids of  claim 36 , wherein the binding protein comprises
 (a) a heavy chain constant region on the first polypeptide chain comprising a human IgG1 heavy chain sequence modified by one or more amino acid changes, wherein the changes comprise substitution of leucines at positions 234 and 235 with alanines, wherein the amino acid positions are numbered using EU index numbering; and   (b) a light chain constant region on the second polypeptide chain comprising a wild type human kappa light chain constant region sequence.   
     
     
         55 . The isolated nucleic acid or group of nucleic acids of  claim 36 , wherein:
 (i) the binding protein is capable of binding IL-1β with a K D  of about 3.4×10 −11  M, as measured by surface plasmon resonance, or capable of inhibiting IL-1β with an IC50 of about 0.018 nM, as measured in an IL-1β neutralization assay, and/or   (ii) the binding protein is capable of binding IL-17 with a K D  of about 4.8×10 −12  M, as measured by surface plasmon resonance, or capable of inhibiting IL-17 with an IC50 of about 0.068 nM, as measured in an IL-17 neutralization assay.   
     
     
         56 . The isolated nucleic acid or group of nucleic acids of  claim 35 , wherein the binding protein comprises any one of:
 DVD2423 (comprising SEQ ID NOs: 48 and 49);   DVD2424 (comprising SEQ ID NOs: 50 and 51);   DVD2425 (comprising SEQ ID NOs: 52 and 53);   DVD2426 (comprising SEQ ID NOs: 54 and 55);   DVD2427 (comprising SEQ ID NOs: 56 and 57);   DVD2428 (comprising SEQ ID NOs: 58 and 59);   DVD2429 (comprising SEQ ID NOs: 60 and 61);   DVD2430 (comprising SEQ ID NOs: 62 and 63);   DVD2431 (comprising SEQ ID NOs: 64 and 65);   DVD2432 (comprising SEQ ID NOs: 66 and 67);   DVD2433 (comprising SEQ ID NOs: 68 and 69);   DVD2434 (comprising SEQ ID NOs: 70 and 71);   DVD2435 (comprising SEQ ID NOs: 72 and 73);   DVD2436 (comprising SEQ ID NOs: 74 and 75);   DVD2437 (comprising SEQ ID NOs: 76 and 77);   DVD2438 (comprising SEQ ID NOs: 78 and 79);   DVD2439 (comprising SEQ ID NOs: 80 and 81);   DVD2440 (comprising SEQ ID NOs: 82 and 83);   DVD2441 (comprising SEQ ID NOs: 84 and 85);   DVD2442 (comprising SEQ ID NOs: 86 and 87);   DVD3415 (comprising SEQ ID NOs: 98 and 99); and   DVD3418 (comprising SEQ ID NOs: 104 and 105).   
     
     
         57 . A vector comprising the isolated nucleic acid or group of nucleic acids of  claim 35 . 
     
     
         58 . The vector of  claim 57 , wherein the isolated nucleic acid or group of nucleic acids encode a binding protein comprising a first polypeptide chain of SEQ ID NO: 104 and a second polypeptide chain of SEQ ID NO: 105. 
     
     
         59 . A host cell comprising the vector of  claim 57 . 
     
     
         60 . The host cell of  claim 59 , wherein the isolated nucleic acid or group of nucleic acids within the vector in the host cell encode a binding protein comprising a first polypeptide chain of SEQ ID NO: 104 and a second polypeptide chain of SEQ ID NO: 105. 
     
     
         61 . The host cell of  claim 59 , wherein the host cell is a prokaryotic cell,  Escherichia coli , a eukaryotic cell, a protist cell, an animal cell, a plant cell, a fungal cell, a yeast cell, an Sf9 cell, a mammalian cell, an avian cell, an insect cell, a CHO cell or a COS cell. 
     
     
         62 . A method of producing a binding protein, comprising culturing the host cell of  claim 59  in culture medium under conditions sufficient to produce the binding protein. 
     
     
         63 . The method of  claim 62 , wherein the binding protein produced by the method has a first polypeptide chain comprising SEQ ID NO: 104 and a second polypeptide chain comprising SEQ ID NO: 105. 
     
     
         64 . A method of detecting the presence, amount, or concentration of IL-1β and/or IL-17 in a test sample by an immunoassay,
 wherein the immunoassay comprises contacting the test sample with at least one binding protein and at least one detectable label, and wherein the at least one binding protein comprises the binding protein produced by the method of  claim 62 . 
 
     
     
         65 . A kit for assaying a test sample for the presence, amount, or concentration of IL-1β and/or IL-17 in the sample, the kit comprising (a) instructions for assaying the test sample for IL-1β and/or IL-17 and (b) at least one binding protein comprising the binding protein produced by the method of  claim 62 .