Antibodies for epidermal growth factor receptor 3 (her3)

Abstract

The present invention relates to antibodies or fragments thereof that target a conformational epitope of a HER receptor. In particular, the invention relates to antibodies or fragments thereof that target a conformational epitope of HER3 receptor and compositions and methods of use thereof.

1 . An isolated antibody or fragment thereof that recognizes a conformational epitope of a HER3 receptor, wherein the conformational epitope comprises amino acid residues within domain 2 and domain 4 of the HER3 receptor, and wherein the antibody or fragment thereof blocks both ligand-dependent and ligand-independent signal transduction. 
     
     
         2 . The isolated antibody or fragment thereof of  claim 1 , wherein the antibody or fragment thereof binds to the inactive state of the HER3 receptor. 
     
     
         3 . The isolated antibody or fragment thereof of  claim 1 , wherein the antibody or fragment thereof stabilizes the HER3 receptor in the inactive state. 
     
     
         4 . The isolated antibody or fragment thereof of  claim 1 , wherein a HER3 ligand can concurrently bind to a ligand binding site on the HER3 receptor. 
     
     
         5 . The isolated antibody or fragment thereof of  claim 4 , wherein the HER3 ligand is selected from the group consisting of neuregulin 1 (NRG), neuregulin 2, betacellulin, heparin-binding epidermal growth factor, and epiregulin. 
     
     
         6 . The isolated antibody of  claim 1 , wherein the conformational epitope comprises amino acid residues 265-277, 315 (of domain 2), 571, 582-584, 596-597, 600-602, 609-615 (of domain 4), or a subset thereof. 
     
     
         7 . The isolated antibody of  claim 1 , wherein the VH of the antibody or fragment thereof binds to at least one of the following HER3 residues: Asn266, Lys267, Leu268, Thr269, Gln271, Glu273, Pro274, Asn275, Pro276, His277, Asn315, Asp571, Pro583, His584, Ala596, Lys597. 
     
     
         8 . The isolated antibody of  claim 1 , wherein the VL of the antibody or fragment thereof binds to at least one of the following HER3 residues: Tyr265, Lys267, Leu268, Phe270, Gly582, Pro583, Lys597, Ile600, Lys602, Glu609, Arg611, Pro612, Cys613, His614, Glu615. 
     
     
         9 . The isolated antibody or fragment thereof of  claim 1 , wherein the antibody is selected from the group consisting of a monoclonal antibody, a polyclonal antibody, a chimeric antibody, a humanized antibody, and a synthetic antibody. 
     
     
         10 . An isolated antibody or fragment thereof that recognizes a conformational epitope of a HER3 receptor, wherein the conformational epitope comprises amino acid residues within domain 2 and domain 4 of the HER3 receptor, wherein binding of the antibody or fragment thereof to the HER3 receptor in the absence of a HER3 ligand reduces ligand-independent formation of a HER2-HER3 protein complex in a cell which expresses HER2 and HER3. 
     
     
         11 . The isolated antibody or fragment thereof of  claim 10 , wherein antibody or fragment thereof stabilizes the HER3 receptor in an inactive state such that the HER3 receptor fails to dimerize with the HER2 receptor to form a HER2-HER3 protein complex. 
     
     
         12 . The isolated antibody or fragment thereof of  claim 11 , wherein the failure to form a HER2-HER3 protein complex prevents activation of signal transduction. 
     
     
         13 . An isolated antibody or fragment thereof that recognizes a conformational epitope of a HER3 receptor, wherein the conformational epitope comprises amino acid residues within domain 2 and domain 4 of the HER3 receptor, wherein binding of the antibody or fragment thereof to the HER3 receptor in the presence of a HER3 ligand reduces ligand-dependent formation of a HER2-HER3 protein complex in a cell which expresses HER2 and HER3. 
     
     
         14 . The isolated antibody or fragment thereof of  claim 13 , wherein antibody or fragment thereof stabilizes the HER3 receptor in an inactive state such that the HER3 receptor fails to dimerize with the HER2 receptor in the presence of a HER3 ligand to form a HER2-HER3 protein complex. 
     
     
         15 . The isolated antibody or fragment thereof of  claim 13 , wherein the failure to form a HER2-HER3 protein complex prevents activation of signal transduction. 
     
     
         16 . The isolated antibody or fragment thereof of  claim 13 , wherein ligand is selected from the group consisting of neuregulin 1 (NRG), neuregulin 2, betacellulin, heparin-binding epidermal growth factor, and epiregulin. 
     
     
         17 . The isolated antibody or fragment thereof of  claim 13 , wherein the antibody or fragment thereof inhibits phosphorylation of HER3 as assessed by a HER3 ligand-independent phosphorylation assay. 
     
     
         18 . The isolated antibody or fragment thereof of  claim 17 , wherein the HER3 ligand-independent phosphorylation assay uses HER2 amplified cells, wherein the HER2 amplified cells are SK-Br-3 cells. 
     
     
         19 . An isolated antibody or fragment thereof that recognizes a conformational epitope of a HER3 receptor, wherein the conformational epitope comprises amino acid residues within domain 2 and domain 4 of the HER3 receptor, and wherein the antibody or fragment thereof inhibits phosphorylation of HER3 as assessed by HER3 ligand-dependent phosphorylation assay. 
     
     
         20 . The isolated antibody or fragment thereof of  claim 19 , wherein the HER3 ligand-dependent phosphorylation assay uses stimulated MCF7 cells in the presence of neuregulin (NRG). 
     
     
         21 . An isolated antibody or fragment thereof to a HER3 receptor, having a dissociation (K D ) of at least 1×10 7 M −1 , 10 8 M −1 , 10 9 M −1 , 10 10  M −1 , 10 11  M −1 , 10 12 M −1 , 10 13  M −1 . 
     
     
         22 . The isolated antibody or fragment thereof of  claim 21 , wherein the antibody or fragment thereof inhibits phosphorylation of HER3 as measured by an in vitro phosphorylation assay selected from the group consisting of phospho-HER3 and phospho-Akt. 
     
     
         23 . An isolated antibody or fragment thereof, binds to the same conformational epitope as an antibody described in Table 1. 
     
     
         24 . An isolated antibody or fragment thereof that cross-competes with an antibody described in Table 1. 
     
     
         25 . An isolated antibody or fragment thereof to HER3 receptor comprising a heavy chain CDR3 selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 10, SEQ ID NO: 22, SEQ ID NO: 28, SEQ ID NO: 40, SEQ ID NO: 46, SEQ ID NO: 58, SEQ ID NO: 64, SEQ ID NO: 76, SEQ ID NO: 82, SEQ ID NO: 94, SEQ ID NO: 100, SEQ ID NO: 112, SEQ ID NO: 118, SEQ ID NO: 130, SEQ ID NO: 136, SEQ ID NO: 148, SEQ ID NO: 166, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 220, SEQ ID NO: 238, SEQ ID NO: 256, SEQ ID NO: 274, SEQ ID NO: 292, SEQ ID NO: 310, SEQ ID NO: 328, SEQ ID NO: 346, and SEQ ID NO: 364. 
     
     
         26 . An isolated antibody or fragment thereof to HER3 receptor which antibody comprises a VH selected from the group consisting of SEQ ID NO: 15, SEQ ID NO: 33, SEQ ID NO: 51, SEQ ID NO: 69, SEQ ID NO: 87, SEQ ID NO: 105, SEQ ID NO: 123, SEQ ID NO: 141, SEQ ID NO: 159, SEQ ID NO: 177, SEQ ID NO: 195, SEQ ID NO: 213, SEQ ID NO: 231, SEQ ID NO: 249, SEQ ID NO: 267, SEQ ID NO: 285, SEQ ID NO: 303, SEQ ID NO: 321, SEQ ID NO: 339, SEQ ID NO: 357, and SEQ ID NO: 375; and a VL selected from the group consisting of SEQ ID NO: 14, SEQ ID NO: 32, SEQ ID NO: 50, SEQ ID NO: 68, SEQ ID NO: 86, SEQ ID NO: 104, SEQ ID NO: 122, SEQ ID NO: 140, SEQ ID NO: 158, SEQ ID NO: 176, SEQ ID NO: 194, SEQ ID NO: 212, SEQ ID NO: 230, SEQ ID NO: 248, SEQ ID NO: 266, SEQ ID NO: 284, SEQ ID NO: 302, SEQ ID NO: 320, SEQ ID NO: 338, SEQ ID NO: 356, and SEQ ID NO: 374 or an amino acid sequence with 97-99% identity thereof. 
     
     
         27 . An isolated antibody or fragment thereof comprising a variable heavy chain sequence having SEQ ID NO: 493. 
     
     
         28 . An isolated antibody or fragment thereof comprising a variable light chain sequence having SEQ ID NO: 494. 
     
     
         29 . An isolated antibody or fragment thereof comprising a variable heavy chain sequence having SEQ ID NO: 493 and a variable light chain sequence having SEQ ID NO: 494. 
     
     
         30 . An isolated antibody or fragment thereof to HER3 receptor with a variant heavy chain variable region comprising CDR1, CDR2, and CDR3, wherein the variant has at least one to four amino acid changes in one of CDR1, CDR2, or CDR3. 
     
     
         31 . An isolated antibody or fragment thereof, comprising a heavy chain variable region CDR1 selected from the group consisting of SEQ ID NO: 2, 20, 38, 56, 74, 92, 110, 128, 146, 164, 182, 200, 218, 236, 254, 272, 290, 308, 326, 344, and 362; CDR2 selected from the group consisting of SEQ ID NO: 3, 21, 39, 57, 75, 93, 111, 129, 147, 165, 183, 201, 219, 237, 255, 273, 291, 309, 327, 345, and 363; CDR3 selected from the group consisting of SEQ ID NO: 4, 22, 40, 58, 76, 94, 112, 130, 148, 166, 184, 202, 220, 238, 256, 274, 292, 310, 328, 346, and 364; a light chain variable region CDR1 selected from the group consisting of SEQ ID NO: 5, 23, 41, 59, 77, 95, 113, 131, 149, 167, 185, 203, 221, 239, 257, 275, 293, 311, 329, 347, and 365; CDR2 selected from the group consisting of SEQ ID NO: 6, 24, 42, 60, 78, 96, 114, 132, 150, 166, 186, 204, 222, 240, 258, 276, 294, 312, 330, 348, and 366; and CDR3 selected from the group consisting of SEQ ID NO: 7, 25, 43, 61, 79, 97, 115, 133, 151, 169, 187, 205, 223, 241, 259, 277, 295, 313, 331, 349, and 367. 
     
     
         32 . An isolated antibody or fragment thereof, comprising a heavy chain variable region CDR1 of SEQ ID NO: 128; CDR2 of SEQ ID NO: 129; CDR3 of SEQ ID NO: 130; a light chain variable region CDR1 of SEQ ID NO: 131; CDR2 of SEQ ID NO: 132; and CDR3 of SEQ ID NO: 133. 
     
     
         33 . A fragment of an antibody that binds to HER3 selected from the group consisting of; Fab, F(ab 2 )′, F(ab) 2 ′, scFv, VHH, VH, VL, dAbs. 
     
     
         34 . A pharmaceutical composition comprising an antibody or fragment thereof and a pharmaceutically acceptable carrier. 
     
     
         35 . The pharmaceutical composition of  claim 34 , further comprising an additional therapeutic agent. 
     
     
         36 . The pharmaceutical composition of  claim 35 , wherein the additional therapeutic agent is selected from the group consisting of an HER1 inhibitor, a HER2 inhibitor, a HER3 inhibitor, a HER4 inhibitor, an mTOR inhibitor and a PI3 Kinase inhibitor. 
     
     
         37 . The pharmaceutical composition of  claim 35 , wherein the additional therapeutic agent is a HER1 inhibitor selected from the group consisting of Matuzumab (EMD72000), Erbitux®/Cetuximab, Vectibix®/Panitumumab, mAb 806, Nimotuzumab, Iressa®/Gefitinib, CI-1033 (PD183805), Lapatinib (GW-572016), Tykerb®/Lapatinib Ditosylate, Tarceva®/Erlotinib HCL (OSI-774), PKI-166, and Tovok®; a HER2 inhibitor selected from the group consisting of Pertuzumab, Trastuzumab, MM-111, neratinib, lapatinib or lapatinib ditosylate/Tykerb®; a HER3 inhibitor selected from the group consisting of, MM-121, MM-111, IB4C3, 2DID12 (U3 Pharma AG), AMG888 (Amgen), AV-203 (Aveo), MEHD7945A (Genentech) and small molecules that inhibit HER3; and a HER4 inhibitor. 
     
     
         38 . The pharmaceutical composition of  claim 35 , wherein the additional therapeutic agent is an mTOR inhibitor selected from the group consisting of Temsirolimus/Torisel®, ridaforolimus/Deforolimus, AP23573, MK8669, everolimus/Affinitor®. 
     
     
         39 . The pharmaceutical composition of  claim 35 , wherein the additional therapeutic agent is a PI3 Kinase inhibitor selected from the group consisting of GDC 0941, BEZ235, BMK120 and BYL719. 
     
     
         40 . A method of treating a cancer comprising selecting a subject having an HER3 expressing cancer, administering to the subject an effective amount of a composition comprising an antibody or fragment thereof disclosed in Table 1. 
     
     
         41 . The method of  claim 40 , wherein the subject is a human and the cancer is selected from the group consisting of breast cancer, colorectal cancer, lung cancer, multiple myeloma, ovarian cancer, liver cancer, gastric cancer, pancreatic cancer, prostate cancer, acute myeloid leukemia, chronic myeloid leukemia, osteosarcoma, squamous cell carcinoma, peripheral nerve sheath tumors, schwannoma, head and neck cancer, bladder cancer, esophageal cancer, glioblastoma, clear cell sarcoma of soft tissue, malignant mesothelioma, neurofibromatosis, renal cancer, and melanoma. 
     
     
         42 . The method of  claim 40 , wherein the cancer is breast cancer.