Tgf-beta binding antibodies

The invention relates to treatment of cell proliferative diseases, disorders, and conditions associated with TGF-β. In particular, the invention provides antibodies and antigen-binding fragments thereof that neutralize mature, human TGF-β1, -β2, and -β3.

The TGF-β protein family consists of three distinct isoforms (TGF-β1, -β2, and -β3) whose pathways activate and regulate multiple gene responses that influence disease states such as, e.g., cell proliferative, inflammatory, and cardiovascular conditions. TGF-β isoform expression in cancer is complex and variable with different combinations of TGF-β isoforms having different roles in particular cancers. For example, TGF-β1 and -β3 play a greater role in ovarian cancer and its progression than TGF-β2; while TGF-β1 and -β2 expression is greater in higher grade chondrosarcoma tumors than -β3. In human breast cancer, TGF-β1 and -β3 are highly expressed, with -β3 expression correlating with overall survival—patients with node metastasis and positive TGF-β3 expression have poor prognostic outcomes. However, in colon cancer, TGF-β1 and -β2 are more highly expressed than -β3 and are present at greater circulating levels than in cancer-free individuals. In glioma cancer, TGF-β2 is pivotal for cell migration. Consequently, there is a need to modulate multiple TGF-β isoform expression in cell proliferation conditions such as cancer.

U.S. Pat. No. 5,571,714 discloses the use of anti-TGF antibodies in treating malignancies and metastatic cancer, and in particular, discloses a murine antibody designated 1D11.16 (ATCC No. HB9849) that is said to bind both TGF-β1 and -β2. This document states that antibody 1D11.16 binds TGF-β2 with a Ka of merely 3.4×10⁸ L/mol (Kd=1/Ka).

The treatment of cell proliferative disorders, such as malignancies and cancers, may be improved by use of antibodies that neutralize mature, human, TGF-β1, -β2, and -β3 with improved binding kinetics and affinity. Substantial physical and chemical stability, adequate pharmacokinetics, and good solubility are also desirable for a pharmaceutical product. Consequently, an unmet need remains for antibodies having characteristics suitable for the pharmaceutical treatment of cell proliferative disorders.

The present invention provides monoclonal antibody compositions that neutralize mature human TGF-β1, -β2 and -β3 with a K_d less than 4 pM for mature human TGF-β1, and a K_d less than 8 pM for mature human TGF-β2, and a K_d less than 4 pM for mature human TGF-β3. The invention also provides antibodies having the following combinations of light chain-(LCVR) and heavy chain-variable region (HCVR) sequences: SEQ ID NO: 27 and 51, 27 and 59, 27 and 60, 27 and 61, 27 and 62, 27 and 63, 27 and 64, 28 and 52, 29 and 51, 30 and 53, 31 and 54, 32 and 55, 33 and 56, 34 and 51, 34 and 57, 35 and 58, 36 and 51, 36 and 69, 36 and 75, 37 and 51, 38 and 51, 39 and 51, 40 and 51, 41 and 64, 41 and 67, 42 and 66, 43 and 68, 44 and 66, 45 and 51, 45 and 69, 46 and 70, 46 and 71, 47 and 71, 48 and 72, 49 and 73, 50 and 65, or 50 and 74. The invention also includes antigen-binding fragments of such antibodies, as well as antibodies and fragments having human or humanized frameworks and constant regions. The antibodies and fragments of the invention are useful to treat cell proliferative diseases, disorders, and conditions.

In one embodiment, antibodies of the present invention neutralize mature human TGF-β1, mature human TGF-β2, and mature human TGF-β3, and have an IC₅₀ of less than or equal to about 100 pM for mature human TGF-β1, and an IC₅₀ of less than or equal to about 400 pM for mature human TGF-β2, and an IC₅₀ of less than or equal to about 200 pM for mature human TGF-β3 in an in vitro HT-2 Cell neutralization assay.

In one embodiment, antibodies of the present invention comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), wherein said HCVR comprises a peptide at CDRH1 with a sequence as shown in SEQ ID NO: 96 or SEQ ID NO: 100, a peptide at CDRH2 with a sequence as shown in SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, or SEQ ID NO: 102, and a peptide at CDRH3 with a sequence as shown in SEQ ID NO: 98, and wherein said LCVR comprises a peptide at CDRL1 with a sequence as shown in SEQ ID NO: 88, SEQ ID NO: 91, SEQ ID NO: 92, or SEQ ID NO: 93, a peptide at CDRL2 with a sequence as shown in SEQ ID NO: 89 or SEQ ID NO: 94, and a peptide at CDRL3 with a sequence as shown in SEQ ID NO: 90, or SEQ ID NO: 95.

In one embodiment, antibodies of the present invention further comprise a human framework region.

In another embodiment, antibodies of the present invention comprise a HCVR with a sequence selected from the group consisting of SEQ ID NO: 64, 68, 69, 70, and 74, and a LCVR with a sequence selected from the group consisting of SEQ ID NO: 41, 43, 45, 46, and 50.

In another embodiment, antibodies of the present invention comprise a HCVR with a sequence as shown in SEQ ID NO: 69, and a LCVR with a sequence as shown in SEQ ID NO: 45.

In one embodiment, antibodies of the present invention comprise a heavy chain with a sequence selected from the group consisting of SEQ ID NO: 77, 79, 81, 83 and 85, and a light chain with a sequence selected from the group consisting of SEQ ID NO: 76, 78, 80, 82, and 84.

In one embodiment, antibodies of the present invention comprise a heavy chain with a sequence as shown in SEQ ID NO: 81 and a light chain with a sequence as shown in SEQ ID NO: 80.

The invention provides a method of treating cell proliferatvie disorders in a mammal, preferably a primate, and more preferably a human, comprising administering to a mammal in need of such treatment an effective amount of an antibody of the invention or a fragment thereof.

The invention provides a method of treating a disease or condition in which fibrogenesis and/or angiogenesis mediated by TGF-β are implicated in a mammal, preferably a primate, and more preferably a human, including myelodysplastic syndrome (MDS)/myeloproliferative disorder (MPD), breast cancer, prostate cancer, ovarian cancer, hepatocellular carcinoma, pancreatic cancer, multiple myeloma, colorectal cancer, other hematological malignancies (hairy cell leukemia, CML, AML, etc), comprising administering to a mammal in need of such treatment an effective amount of an antibody of the invention or a fragment thereof. This may further comprise administering to the mammal an effective amount of a therapeutic agent other than anti-TGF-β antibodies, such as a chemotherapeutic agent, anti-angiogenic agent, or cytotoxic chemotherapy.

In another embodiment, this invention provides a method of treating a patient with HER2-overexpressing metastatic breast cancer, comprising administering to the patent an effective amount of an antibody of the invention and an antibody that blocks signaling through the HER-2 receptor, i.e. trastuzumab.

The present invention also provides a pharmaceutical composition comprising an antibody of the invention in combination with a pharmaceutically acceptable carrier, diluent or excipient.

This invention also provides the use of an antibody of the invention for the manufacture of a medicament for treating cell proliferatvie disorders in mammals. Additionally, this invention provides a pharmaceutical composition adapted for treating cell proliferatvie disorders comprising an antibody of the invention in combination with one or more pharmaceutically acceptable excipients, carriers, or diluents thereof The invention also provides the use of an antibody of the invention for the manufacture of a medicament for treating a disease or condition capable of being improved or prevented by neutralizing TGF-β activities.

This invention also provides the use of an antibody of the invention for the manufacture of a medicament for the treatment of a disease or condition in which fibrogenesis and/or angiogenesis mediated by TGF-β are implicated in a mammal, preferably a primate, and more preferably a human, including MDS/MPD, breast cancer, prostate cancer, ovarian cancer, hepatocellular carcinoma, pancreatic cancer, multiple myeloma, colorectal cancer, other hematological malignancies (hairy cell leukemia, CML, AML, etc), comprising administering to a mammal in need of such treatment an effective amount of an antibody of the invention or a fragment thereof. This may further comprise administering to the mammal an effective amount of a therapeutic agent other than anti-TGF-β antibodies, such as a chemotherapeutic, anti-angiogenic, or cytotoxic agent, or a cytokine. Furthermore, this invention provides a pharmaceutical composition adapted for the treatment of conditions in which neutralization or reduction of TGF-β activities would be beneficial in a mammal, including MDS/MPD, breast cancer, prostate cancer, ovarian cancer, hepatocellular carcinoma, pancreatic cancer, multiple myeloma, colorectal cancer, other hematological malignancies (hairy cell leukemia, CML, AML, etc), comprising an antibody of the invention in combination with one or more pharmaceutically acceptable excipients, carriers, or diluents. The invention also provides the use of an antibody of the invention for the manufacture of a medicament for treating a disease or condition capable of being improved or prevented by neutralizing or reducing TGF-β activities.