ANTI-TNF/IL-17 DUAL VARIABLE DOMAIN IMMUNOGLOBULIN AND USES THEREOF
The present invention relates to engineered multivalent and multispecific binding proteins, methods of making, and specifically to their uses in the prevention and/or treatment of acute and chronic inflammatory and other diseases.
1 . A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first variable domain, VD2 is a second variable domain, C is a constant domain, X1 represents an amino acid or polypeptide, X2 represents an Fc region and n is 0 or 1, wherein the binding protein is capable of binding TNF and IL-17.
2 . The binding protein of claim 1 , wherein the VD1 and the VD2 are heavy chain variable domains.
3 - 5 . (canceled)
6 . The binding protein of claim 2 , wherein C is a heavy chain constant domain.
7 . The binding protein of claim 6 , wherein X1 is a linker with the proviso that X1 is not CH1.
8 . The binding protein of claim 7 , wherein the linker is AKTTPKLEEGEFSEAR; AKTTPKLEEGEFSEARV; AKTTPKLGG; SAKTTPKLGG; AKTTPKLEEGEFSEARV; SAKTTP; SAKTTPKLGG; RADAAP; RADAAPTVS; RADAAAAGGPGS; RADAAAA(G 4 S) 4 , SAKTTP; SAKTTPKLGG; SAKTTPKLEEGEFSEARV; ADAAP; ADAAPTVSIFPP; TVAAP; TVAAPSVFIFPP; QPKAAP; QPKAAPSVTLFPP; AKTTPP; AKTTPPSVTPLAP; AKTTAP; AKTTAPSVYPLAP; ASTKGP; ASTKGPSVFPLAP; GGGGSGGGGSGGGGS; GENKVEYAPALMALS; GPAKELTPLKEAKVS; or GHEAAAVMQVQYPAS.
9 - 10 . (canceled)
11 . A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, 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 with the proviso that it is not CH1, and X2 is an Fc region, wherein the binding protein is capable of binding TNF and IL-17.
12 . The binding protein of claim 11 , wherein the VD1 and the VD2 are light chain variable domains.
13 - 15 . (canceled)
16 . The binding protein of claim 12 , wherein C is a light chain constant domain.
17 . The binding protein of claim 16 , wherein X1 is a linker with the proviso that X1 is not CL1.
18 . The binding protein of claim 17 , wherein the linker is AKTTPKLEEGEFSEAR; AKTTPKLEEGEFSEARV; AKTTPKLGG; SAKTTPKLGG; AKTTPKLEEGEFSEARV; SAKTTP; SAKTTPKLGG; RADAAP; RADAAPTVS; RADAAAAGGPGS; RADAAAA(G 4 S) 4 , SAKTTP; SAKTTPKLGG; SAKTTPKLEEGEFSEARV; ADAAP; ADAAPTVSIFPP; TVAAP; TVAAPSVFIFPP; QPKAAP; QPKAAPSVTLFPP; AKTTPP; AKTTPPSVTPLAP; AKTTAP; AKTTAPSVYPLAP; ASTKGP; ASTKGPSVFPLAP; GGGGSGGGGSGGGGS; GENKVEYAPALMALS; GPAKELTPLKEAKVS; or GHEAAAVMQVQYPAS.
19 . The binding protein of claim 17 , wherein the binding protein does not comprise X2.
20 . A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, 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 with the proviso that it is not CH1, and X2 does not comprise an Fc region, wherein the binding protein is capable of binding TNF and IL-17.
21 . A binding protein comprising four polypeptide chains, wherein:
a first polypeptide chain and a second polypeptide chain each comprises VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first variable domain, VD2 is a second variable domain, C is a constant domain, X1 is a linker with the proviso that it is not a constant domain, X2 is an Fc region, and n is 0 or 1; and
a third polypeptide chain and a fourth polypeptide chain each comprises VD1-(X1)n-VD2-C, wherein VD1 is a first variable domain, VD2 is a second variable domain, C is a constant domain, and X1 is a linker with the proviso that it is not a constant domain, and n is 0 or 1;
wherein either or each of VD1 and VD2 of the polypeptide chains is a non-immunoglobulin variable domain; and
wherein the four polypeptide chains of the binding protein form four functional binding domains, wherein said binding protein is capable of binding TNF and IL-17.
22 - 28 . (canceled)
29 . The binding protein of claim 21 , wherein X1 is AKTTPKLEEGEFSEAR; AKTTPKLEEGEFSEARV; AKTTPKLGG; RADAAP; RADAAPTVS; RADAAAAGGPGS; RADAAAA(G 4 S) 4 , SAKTTP; SAKTTPKLGG; SAKTTPKLEEGEFSEARV; ADAAP; ADAAPTVSIFPP; TVAAP; TVAAPSVFIFPP; QPKAAP; QPKAAPSVTLFPP; AKTTPP; AKTTPPSVTPLAP; AKTTAP; AKTTAPSVYPLAP; ASTKGP; ASTKGPSVFPLAP; GGGGSGGGGSGGGGS; GENKVEYAPALMALS; GPAKELTPLKEAKVS; or GHEAAAVMQVQYPAS.
30 . The binding protein of claim 21 , wherein, for (X2)n, n is 0.
31 - 41 . (canceled)
42 . An isolated nucleic acid comprising a coding sequence for a polypeptide chain comprising the structure VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a variable domain, VD2 is a second variable domain, C is a constant domain, X1 is a linker with the proviso that it is not a constant domain, X2 is an Fc region, and n is 0 or 1, wherein the binding protein is capable of binding TNF and IL-17.
43 . (canceled)
44 . (canceled)
45 . A vector comprising the nucleic acid of claim 42 .
46 - 47 . (canceled)
48 . A host cell comprising a vector of claim 45 .
49 - 68 . (canceled)
69 . A method of producing a four-polypeptide chain binding protein having four functional antigen binding sites comprising:
a) transfecting a host cell with a first expression vector and a second expression vector, wherein:
the first expression vector comprises a nucleic acid comprising a coding sequence for a first polypeptide chain comprising VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first immunoglobulin heavy chain variable domain, VD2 is a second immunoglobulin heavy chain variable domain, C is a constant domain, X1 is a linker with the proviso that it is not a constant domain, X2 is an Fc region, and n is 0 or 1; and
the second expression vector comprises a nucleic acid comprising a coding sequence for a second polypeptide chain comprising VD1-(X1)n-VD2-C, wherein VD1 is a first immunoglobulin light chain variable domain, VD2 is a second immunoglobulin light chain variable domain, C is a constant domain, X1 is a linker with the proviso that it is not a constant domain, and n is 0 or 1; and
b) culturing the transfected host cell in (a) in culture medium under conditions sufficient to produce a four polypeptide chain binding protein comprising two first polypeptide chains in association with two second polypeptide chains, wherein the four polypeptide chains of the binding protein form four functional antigen binding domains, and wherein the binding protein is capable of binding TNF and IL-17.
70 - 72 . (canceled)
73 . A method for treating a subject for a disease or a disorder by administering to the subject a binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first variable domain, VD2 is a second variable domain, C is a constant domain, X1 represents an amino acid or polypeptide, X2 represents an Fc region and n is 0 or 1, wherein the binding protein is capable of binding TNF and IL-17, such that treatment is achieved.
74 . (canceled)
75 . (canceled)