ANTIBODIES THAT BIND IL-4 AND/OR IL-13 AND THEIR USES
The present invention relates to novel humanized anti-IL-4 and IL-13 antibodies and fragments thereof and novel bispecific antibodies and fragments thereof that specifically bind to IL-4 and IL-13. The invention also includes uses of the antibodies to treat or prevent IL-4 and/or IL-13 mediated diseases or disorders, including allergic asthma and dermatitis.
1 - 49 . (canceled)
50 . A bispecific antibody or bispecific antibody fragment thereof that specifically binds to IL-13 and IL-4;
wherein the bispecific antibody or bispecific antibody fragment thereof comprises two light chains having the structure V L1 -L 1 -V L2 -C L , and two heavy chains having the structure V H1 -L 2 -V H2 -C H1 ;
wherein C L is a constant light chain domain and C H1 is a constant heavy chain domain;
wherein L 1 and L 2 are amino acid linkers; and
wherein:
(a) V L1 comprises the amino acid sequence of SEQ ID NO: 1, V L2 comprises the amino acid sequence of SEQ ID NO: 3, V H1 comprises the amino acid sequence of SEQ ID NO: 2, and V H2 comprises the amino acid sequence of SEQ ID NO: 4;
(b) V L1 comprises the amino acid sequence of SEQ ID NO: 1, V L2 comprises the amino acid sequence of SEQ ID NO: 3, V H1 comprises the amino acid sequence of SEQ ID NO: 2, and V H2 comprises the amino acid sequence of SEQ ID NO: 5;
(c) V L1 comprises the amino acid sequence of SEQ ID NO: 3, V L2 comprises the amino acid sequence of SEQ ID NO: 1, V H1 comprises the amino acid sequence of SEQ ID NO: 4, and V H2 comprises the amino acid sequence of SEQ ID NO: 2;
(d) V L1 comprises the amino acid sequence of SEQ ID NO: 3, V L2 comprises the amino acid sequence of SEQ ID NO: 1, V H1 comprises the amino acid sequence of SEQ ID NO: 5, and V H2 comprises the amino acid sequence of SEQ ID NO: 2;
(e) V L1 comprises the amino acid sequences RASESVDSYGQSYMH (SEQ ID NO: 8), LASNLES (SEQ ID NO: 9), and QQNAEDSRT (SEQ ID NO: 10), V L2 comprises the amino acid sequences HASQNIDVWLS (SEQ ID NO: 14), KASNLHTG (SEQ ID NO: 15), and QQAHSYPFT (SEQ ID NO: 16), V H1 comprises the amino acid sequences GFSLTDSSIN (SEQ ID NO: 11), DGRID (SEQ ID NO: 12), and DGYFPYAMDF (SEQ ID NO: 13), and V H2 comprises the amino acid sequences GYSFTSYWIH (SEQ ID NO: 17), IDPSDGETR (SEQ ID NO: 18), and LKEYGNYDSFYFDV (SEQ ID NO: 19);
(f) V L1 comprises the amino acid sequences RASESVDSYGQSYMH (SEQ ID NO: 8), LASNLES (SEQ ID NO: 9), and QQNAEDSRT (SEQ ID NO: 10), V L2 comprises the amino acid sequences HASQNIDVWLS (SEQ ID NO: 14), KASNLHTG (SEQ ID NO: 15), and QQAHSYPFT (SEQ ID NO: 16), V H1 comprises the amino acid sequences GFSLTDSSIN (SEQ ID NO: 11), DGRID (SEQ ID NO: 12), and DGYFPYAMDF (SEQ ID NO: 13), and V H2 comprises the amino acid sequences GYSFTSYWIH (SEQ ID NO: 20), IDASDGETR (SEQ ID NO: 21), and LKEYGNYDSFYFDV (SEQ ID NO: 22);
(g) V L1 comprises the amino acid sequences HASQNIDVWLS (SEQ ID NO: 14), KASNLHTG (SEQ ID NO: 15), and QQAHSYPFT (SEQ ID NO: 16), comprises the amino acid sequences RASESVDSYGQSYMH (SEQ ID NO: 8), LASNLES (SEQ ID NO: 9), and QQNAEDSRT (SEQ ID NO: 10), V H1 comprises the amino acid sequences GYSFTSYWIH (SEQ ID NO: 17), IDPSDGETR (SEQ ID NO: 18), and LKEYGNYDSFYFDV (SEQ ID NO: 19), and V H2 comprises the amino acid sequences GFSLTDSSIN (SEQ ID NO: 11), DGRID (SEQ ID NO: 12), and DGYFPYAMDF (SEQ ID NO: 13); or
(h) V L1 comprises the amino acid sequences HASQNIDVWLS (SEQ ID NO: 14), KASNLHTG (SEQ ID NO: 15), and QQAHSYPFT (SEQ ID NO: 16), comprises the amino acid sequences RASESVDSYGQSYMH (SEQ ID NO: 8), LASNLES (SEQ ID NO: 9), and QQNAEDSRT (SEQ ID NO: 10), V H1 comprises the amino acid sequences GYSFTSYWIH (SEQ ID NO: 20), IDASDGETR (SEQ ID NO: 21), and LKEYGNYDSFYFDV (SEQ ID NO: 22), and V H2 comprises the amino acid sequences GFSLTDSSIN (SEQ ID NO: 11), DGRID (SEQ ID NO: 12), and DGYFPYAMDF (SEQ ID NO: 13).
51 . (canceled)
52 . The bispecific antibody or bispecific antibody fragment thereof of either claim 50 or 69 that further comprises additional constant region domains.
53 . The bispecific antibody or bispecific antibody fragment thereof of claim 52 , wherein the additional constant region domains consist of C H2 and C H3 .
54 . The bispecific antibody or bispecific antibody fragment thereof of claim 50 , wherein L 1 and L 2 consist of the amino acid sequence of SEQ ID NO: 6.
55 . The bispecific antibody or bispecific antibody fragment thereof of claim 50 , wherein either L 1 and/or L 2 are absent.
56 . The bispecific antibody or bispecific antibody fragment thereof of either claim 50 or 69 that further comprises a label, wherein the label is a radiolabel, fluorophore, chromophore, imaging agent, or metal ion.
57 . The bispecific antibody or bispecific antibody fragment thereof of either claim 50 or 69 that is further conjugated to an effector molecule.
58 . The bispecific antibody or bispecific antibody fragment thereof of claim 57 , wherein the effector molecule is a heterologous polypeptide, drug, radionucleotide, or toxin.
59 . The bispecific antibody or bispecific antibody fragment thereof of either claim 50 or 69 , wherein the bispecific antibody or bispecific antibody fragment thereof neutralizes IL-4 and/or IL-13 activity and preferably neutralizes IL-4 or IL-13 mediated cell proliferation in TF-1 cells.
60 . A nucleic acid molecule encoding the bispecific antibody or bispecific antibody fragment thereof of either claim 50 or 69 .
61 . A vector comprising the nucleic acid molecule of claim 60 .
62 . A host cell comprising the vector of claim 61 .
63 . A pharmaceutical composition comprising the bispecific antibody or bispecific antibody fragment thereof of either claim 50 or 69 and a pharmaceutically acceptable carrier.
64 - 67 . (canceled)
68 . A kit comprising the bispecific antibody or bispecific antibody fragment thereof of either claim 50 or 69 and instructions for use.
69 . A bispecific antibody or bispecific antibody fragment thereof that specifically binds to IL-13 and IL-4,
wherein the bispecific antibody or bispecific antibody fragment thereof comprises two light chains and two heavy chains;
wherein the light chains comprise an outer variable light chain domain linked to an inner variable light chain domain which is linked to a constant light chain domain (CL), and the heavy chains comprise an outer variable heavy chain domain linked to an inner variable heavy chain domain which is linked to a constant heavy chain domain (CH1); and
wherein:
(a) the outer variable light chain domain comprises the amino acid sequence of SEQ ID NO: 3, the inner variable light chain domain comprises the amino acid sequence of SEQ ID NO: 1, the outer variable heavy chain domain comprises the amino acid sequence of SEQ ID NO: 4, and the inner variable heavy chain domain comprises the amino acid sequence of SEQ ID NO: 2;
(b) the outer variable light chain domain comprises the amino acid sequence of SEQ ID NO: 3, the inner variable light chain domain comprises the amino acid sequence of SEQ ID NO: 1, the outer variable heavy chain domain comprises the amino acid sequence of SEQ ID NO: 5, and the inner variable heavy chain domain comprises the amino acid sequence of SEQ ID NO: 2;
(c) the outer variable light chain domain comprises the amino acid sequences HASQNIDVWLS (SEQ ID NO: 14), KASNLHTG (SEQ ID NO: 15), and QQAHSYPFT (SEQ ID NO: 16), the inner variable light chain domain comprises the amino acid sequences RASESVDSYGQSYMH (SEQ ID NO: 8), LASNLES (SEQ ID NO: 9), and QQNAEDSRT (SEQ ID NO: 10), the outer variable heavy chain domain comprises the amino acid sequences GYSFTSYWIH (SEQ ID NO: 17), IDPSDGETR (SEQ ID NO: 18), and LKEYGNYDSFYFDV (SEQ ID NO: 19), and the inner variable heavy chain domain comprises the amino acid sequences GFSLTDSSIN (SEQ ID NO: 11), DGRID (SEQ ID NO: 12), and DGYFPYAMDF (SEQ ID NO: 13); or
(d) the outer variable light chain domain comprises the amino acid sequences HASQNIDVWLS (SEQ ID NO: 14), KASNLHTG (SEQ ID NO: 15), and QQAHSYPFT (SEQ ID NO: 16), the inner variable light chain domain comprises the amino acid sequences RASESVDSYGQSYMH (SEQ ID NO: 8), LASNLES (SEQ ID NO: 9), and QQNAEDSRT (SEQ ID NO: 10), the outer variable heavy chain domain comprises the amino acid sequences GYSFTSYWIH (SEQ ID NO: 20), IDASDGETR (SEQ ID NO: 21), and LKEYGNYDSFYFDV (SEQ ID NO: 22), and the inner variable heavy chain domain comprises the amino acid sequences GFSLTDSSIN (SEQ ID NO: 11), DGRID (SEQ ID NO: 12), and DGYFPYAMDF (SEQ ID NO: 13).