Combination of LIF inhibitors and platinum-based anticancer agents for use in the treatment of cancer - Patent Application 20070229633

Abstract

To provide methods for treating or preventing a cancer, tumors or other neoplasms in individuals.SOLUTION: A method of treating an individual with a cancer comprises administering to the individual with the cancer an effective amount of: a) a Leukemia Inhibitory Factor (LIF) binding polypeptide; and b) a platinum-based antineoplastic agent.SELECTED DRAWING: Figure 18

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of European Patent Application No. 18382432.5, filed June 18, 2018, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] Leukemia inhibitory factor (LIF) is an interleukin-6 (IL-6)-type cytokine involved in various biological activities, including the inhibition of cell differentiation. Human LIF is a 202-amino acid polypeptide that exerts its biological effects through binding to the cell surface LIF receptor (LIFR or CD118), which heterodimerizes with gp130. Human LIF leads to the activation of pro-growth signaling pathways, such as the mitogen-activated protein kinase (MAPK) and Janus-activated kinase (JAK / STAT) pathways. High expression levels and elevated serum levels of LIF have been demonstrated to be associated with poor prognosis for many types of cancer.

[0003] Platinum-based antineoplastic agents, also known as platins, are coordination complexes of platinum that have shown utility in the treatment of several cancers. Mechanistically, platinum-based antineoplastic agents cause cell death by crosslinking or intercalating DNA. Platinum-based antineoplastic agents inhibit DNA repair and synthesis in cells. Ultimately, these cellular processes are impaired, leading to the initiation of apoptosis. Common platinum-based antineoplastic agents can include cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenathriplatin, picoplatin, satraplatin, lobaplatin, or heptaplatin. Summary of the Invention [Means for solving the problem]

[0004] Described herein are methods for treating or preventing cancer, tumors, or other neoplasms in an individual. The methods and compositions in question involve the combination of a LIF-binding polypeptide with a platinum-based antineoplastic agent. These methods can utilize an anti-LIF antibody that antagonizes or blocks LIF activity and the platinum-based antineoplastic agent. Notably, these combinations exhibit surprising synergy when compared to either the anti-LIF antibody or the platinum-based antineoplastic agent alone.

[0005] In one aspect, described herein is the use of a leukemia inhibitory factor (LIF)-binding polypeptide in combination with a platinum-based antineoplastic agent to treat cancer in an individual. In certain embodiments, the LIF-binding polypeptide and the platinum-based antineoplastic agent are administered to the individual in separate formulations. In certain embodiments, the LIF-binding polypeptide and the platinum-based antineoplastic agent are administered to the individual in the same formulation. In certain embodiments, the LIF-binding polypeptide is administered to the individual before the platinum-based antineoplastic agent is administered to the individual. In certain embodiments, the platinum-based antineoplastic agent is administered to the individual before the LIF-binding polypeptide is administered to the individual. In certain embodiments, the platinum-based antineoplastic agent is administered to the individual simultaneously with the LIF-binding polypeptide. In certain embodiments, the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. In certain embodiments, the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. In certain embodiments, an antibody that specifically binds to LIF comprises at least one framework region derived from a human antibody framework region. In certain embodiments, an antibody that specifically binds to LIF is humanized. In certain embodiments, an antibody that specifically binds to LIF is deimmunized. In certain embodiments, an antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. In certain embodiments, an antibody that specifically binds to LIF is an IgG antibody. In certain embodiments, an antibody that specifically binds to LIF is a Fab, F(ab)2, single domain antibody, single chain variable fragment (scFv), or nanobody.In certain embodiments, an antibody that specifically binds to LIF comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; (d) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13. In certain embodiments, an antibody that specifically binds LIF comprises (a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and (b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO:42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO:46.In certain embodiments, an antibody that specifically binds to LIF comprises (a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60 or 67; and (b) an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. In certain embodiments, an antibody that specifically binds to LIF has a K of less than about 200 picomolar. D In certain embodiments, antibodies that specifically bind to LIF have a K of less than about 100 picomolar. D In certain embodiments, the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriplatin, picoplatin, satraplatin, or a combination thereof. In certain embodiments, the platinum-based anti-cancer agent is cisplatin. In certain embodiments, the cancer comprises advanced solid tumors, glioblastoma, gastric cancer, skin cancer, prostate cancer, pancreatic cancer, breast cancer, testicular cancer, thyroid cancer, head and neck cancer, liver cancer, kidney cancer, esophageal cancer, ovarian cancer, colon cancer, lung cancer, lymphoma, or soft tissue cancer. In certain embodiments, the cancer comprises non-small cell lung cancer, ovarian epithelial cancer, or pancreatic adenocarcinoma. In certain embodiments, the cancer is resistant to treatment with a therapeutic amount of the LIF-binding polypeptide or platinum-based anti-cancer agent administered as monotherapy.

[0006] In another aspect, described herein is the use of an antibody that specifically binds leukemia inhibitory factor (LIF) in combination with a platinum-based anti-cancer agent to treat cancer in an individual, wherein the LIF-binding antibody comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1-3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR4) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6-8; (d) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13.

[0007] In another aspect, described herein are methods of treating an individual with cancer, comprising administering to the individual with cancer effective amounts of: (a) a leukemia inhibitory factor (LIF)-binding polypeptide; and (b) a platinum-based anti-cancer agent. In certain embodiments, the method comprises administering to the individual with cancer an effective amount of the LIF-binding polypeptide. In certain embodiments, the method comprises administering to the individual with cancer an effective amount of the platinum-based anti-cancer agent. In certain embodiments, the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. In certain embodiments, the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. In certain embodiments, the antibody that specifically binds to LIF comprises at least one framework region derived from a human antibody framework region. In certain embodiments, the antibody that specifically binds to LIF is humanized. In certain embodiments, the antibody that specifically binds to LIF is deimmunized. In certain embodiments, the antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. In certain embodiments, the antibody that specifically binds to LIF is an IgG antibody. In certain embodiments, the antibody that specifically binds to LIF is a Fab, F(ab)2, single domain antibody, single chain variable fragment (scFv), or nanobody. In certain embodiments, an antibody that specifically binds to LIF comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; (d) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13.In certain embodiments, an antibody that specifically binds LIF comprises (a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and (b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, an antibody that specifically binds to LIF comprises (a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60 or 67; and an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. In certain embodiments, an antibody that specifically binds to LIF has a K of less than about 200 picomolar. D In certain embodiments, antibodies that specifically bind to LIF have a K of less than about 100 picomolar. DIn certain embodiments, the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriplatin, picoplatin, satraplatin, or a combination thereof. In certain embodiments, the platinum-based anti-cancer agent is cisplatin. In certain embodiments, the cancer comprises advanced solid tumors, glioblastoma, gastric cancer, skin cancer, prostate cancer, pancreatic cancer, breast cancer, testicular cancer, thyroid cancer, head and neck cancer, liver cancer, kidney cancer, esophageal cancer, ovarian cancer, colon cancer, lung cancer, lymphoma, or soft tissue cancer. In certain embodiments, the cancer comprises non-small cell lung cancer, ovarian epithelial cancer, or pancreatic adenocarcinoma. In certain embodiments, the cancer is resistant to treatment with a therapeutic amount of an inhibitor of the LIF-binding polypeptide. In certain embodiments, the cancer is resistant to treatment with a therapeutic amount of a platinum-based anti-cancer agent. In certain embodiments, the leukemia inhibitory factor (LIF)-binding polypeptide and the platinum-based anti-neoplastic agent are administered separately. In certain embodiments, the LIF-binding polypeptide and the platinum-based anti-neoplastic agent are administered simultaneously. In certain embodiments, the LIF-binding polypeptide and the platinum-based anti-neoplastic agent are administered in a single composition.

[0008] In another aspect, described herein are methods of treating an individual with cancer, comprising administering an effective amount of a leukemia inhibitory factor (LIF)-binding polypeptide to the individual with cancer, the individual receiving a therapeutic amount of a platinum-based anti-cancer agent. In certain embodiments, the method inhibits cancer growth or metastasis. In certain embodiments, the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. In certain embodiments, the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. In certain embodiments, the LIF-binding polypeptide comprises at least one framework region derived from a human immunoglobulin framework region. In certain embodiments, the antibody that specifically binds to LIF is humanized. In certain embodiments, the antibody that specifically binds to LIF is deimmunized. In certain embodiments, the antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. In certain embodiments, the antibody that specifically binds to LIF is an IgG antibody. In certain embodiments, the antibody that specifically binds to LIF is a Fab, F(ab)2, single domain antibody, single chain variable fragment (scFv), or nanobody. In certain embodiments, an antibody that specifically binds to LIF comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; (d) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13.In certain embodiments, an antibody that specifically binds LIF comprises (a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and (b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, an antibody that specifically binds to LIF comprises (a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60 or 67; and (b) an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. In certain embodiments, an antibody that specifically binds to LIF has a K of less than about 200 picomolar. D In certain embodiments, antibodies that specifically bind to LIF have a K of less than about 100 picomolar. DIn certain embodiments, the platinum-based anti-cancer agent binds at the following structure: In certain embodiments, the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriptin, picoplatin, satraplatin, or a combination thereof. In certain embodiments, the platinum-based anti-cancer agent is cisplatin. In certain embodiments, the cancer comprises advanced solid tumors, glioblastoma, gastric cancer, skin cancer, prostate cancer, pancreatic cancer, breast cancer, testicular cancer, thyroid cancer, head and neck cancer, liver cancer, kidney cancer, esophageal cancer, ovarian cancer, colon cancer, lung cancer, lymphoma, or soft tissue cancer. In certain embodiments, the cancer comprises non-small cell lung cancer, ovarian epithelial cancer, or pancreatic adenocarcinoma. In certain embodiments, the cancer is resistant to treatment with a therapeutic amount of a platinum-based anti-cancer agent.

[0009] In another aspect, described herein are methods of treating an individual with cancer, comprising administering an effective amount of a platinum-based anti-neoplastic agent to the individual with cancer, the individual having cancer and having received a therapeutic amount of a leukemia inhibitory factor (LIF)-binding polypeptide. In certain embodiments, the method inhibits cancer growth or metastasis. In certain embodiments, the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. In certain embodiments, the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. In certain embodiments, the LIF-binding polypeptide comprises at least one framework region derived from a human immunoglobulin framework region. In certain embodiments, the antibody that specifically binds to LIF is humanized. In certain embodiments, the antibody that specifically binds to LIF is deimmunized. In certain embodiments, the antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. In certain embodiments, the antibody that specifically binds to LIF is an IgG antibody. In certain embodiments, the antibody that specifically binds to LIF is a Fab, F(ab)2, single domain antibody, single chain variable fragment (scFv), or nanobody. In certain embodiments, an antibody that specifically binds to LIF comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; (d) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13.In certain embodiments, an antibody that specifically binds LIF comprises (a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and (b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, an antibody that specifically binds to LIF comprises (a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60 or 67; and (b) an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. In certain embodiments, an antibody that specifically binds to LIF has a K of less than about 200 picomolar. D In certain embodiments, antibodies that specifically bind to LIF have a K of less than about 100 picomolar. DIn certain embodiments, the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriplatin, picoplatin, satraplatin, or a combination thereof. In certain embodiments, the platinum-based anti-cancer agent is cisplatin. In certain embodiments, the cancer comprises advanced solid tumors, glioblastoma, gastric cancer, skin cancer, prostate cancer, pancreatic cancer, breast cancer, testicular cancer, thyroid cancer, head and neck cancer, liver cancer, kidney cancer, esophageal cancer, ovarian cancer, colon cancer, lung cancer, lymphoma, or soft tissue cancer. In certain embodiments, the cancer comprises non-small cell lung cancer, ovarian epithelial cancer, or pancreatic adenocarcinoma. In certain embodiments, the cancer is resistant to treatment with a therapeutic amount of an inhibitor of the LIF-binding polypeptide.

[0010] In another aspect, described herein is a method of treating an individual with cancer, comprising administering to the individual an effective amount of: (a) (i) immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1-3; (ii) immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (iii) immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6-8; (iv) immunoglobulin heavy chain complementarity determining region 4 (VH-CDR5) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9-11; (v) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (vi) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13; and (b) a platinum-based anti-cancer agent to an individual with cancer.

[0011] In another aspect, described herein are kits comprising: (a) a leukemia inhibitory factor (LIF)-binding polypeptide; and (b) a platinum-based anti-neoplastic agent. In certain embodiments, the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. In certain embodiments, the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. In certain embodiments, the LIF-binding polypeptide comprises at least one framework region derived from a human immunoglobulin framework region. In certain embodiments, the antibody that specifically binds to LIF is humanized. In certain embodiments, the antibody that specifically binds to LIF is deimmunized. In certain embodiments, the antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. In certain embodiments, the antibody that specifically binds to LIF is an IgG antibody. In certain embodiments, the antibody that specifically binds to LIF is a Fab, F(ab)2, single-domain antibody, single-chain variable fragment (scFv), or nanobody. In certain embodiments, an antibody that specifically binds to LIF comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; (d) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13.In certain embodiments, an antibody that specifically binds LIF comprises (a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and (b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, an antibody that specifically binds to LIF comprises (a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60 or 67; and (b) an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. In certain embodiments, an antibody that specifically binds to LIF has a K of less than about 200 picomolar. D In certain embodiments, antibodies that specifically bind to LIF have a K of less than about 100 picomolar. D In certain embodiments, the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriplatin, picoplatin, satraplatin, or a combination thereof. In certain embodiments, the platinum-based anti-cancer agent is cisplatin. In certain embodiments, the kit further comprises a pharmaceutically acceptable excipient, carrier, or diluent.

[0012] In another aspect, described herein are compositions comprising (a) a leukemia inhibitory factor (LIF)-binding polypeptide; and (b) a platinum-based anti-neoplastic agent. In certain embodiments, the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. In certain embodiments, the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. In certain embodiments, the LIF-binding polypeptide comprises at least one framework region derived from a human immunoglobulin framework region. In certain embodiments, the antibody that specifically binds to LIF is humanized. In certain embodiments, the antibody that specifically binds to LIF is deimmunized. In certain embodiments, the antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. In certain embodiments, the antibody that specifically binds to LIF is an IgG antibody. In certain embodiments, the antibody that specifically binds to LIF is a Fab, F(ab)2, single-domain antibody, single-chain variable fragment (scFv), or nanobody. In certain embodiments, an antibody that specifically binds to LIF comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; (d) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13.In certain embodiments, an antibody that specifically binds LIF comprises (a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and (b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, an antibody that specifically binds to LIF comprises (a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60 or 67; and (b) an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. In certain embodiments, an antibody that specifically binds to LIF has a K of less than about 200 picomolar. D In certain embodiments, antibodies that specifically bind to LIF have a K of less than about 100 picomolar. D In certain embodiments, the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriplatin, picoplatin, satraplatin, or a combination thereof. In certain embodiments, the platinum-based anti-cancer agent is cisplatin. In certain embodiments, the composition further comprises a pharmaceutically acceptable excipient, carrier, or diluent. [Brief explanation of the drawings]

[0013] [Figure 1] FIG. 1 depicts a Western blot showing the inhibition of LIF-induced STAT3 phosphorylation by different anti-LIF humanized antibodies. [Figure 2] 2A-2B depict Western blots showing the inhibition of LIF-induced STAT3 phosphorylation by the humanized parental 5D8 antibody. [Figure 3] Figure 3A shows the IC50 for LIF inhibition in U-251 cells using the h5D8 antibody. Figure 3B shows representative IC50 dose-response curves for r5D8 and h5D8 inhibition of pSTAT3 under endogenous LIF stimulation conditions. Shown are representative curves (n=1 h5D8, n=2 r5D8). [Figure 4] FIG. 4 depicts a Western blot showing the inhibition of LIF-induced STAT3 phosphorylation by different monoclonal antibodies described in this disclosure. [Figure 5] Figure 5 shows immunohistochemical staining and quantification of LIF expression in glioblastoma multiforme (GBM), NSCLC (non-small cell lung cancer), ovarian cancer, colorectal cancer, and pancreatic tumors from human patients. Bars represent the mean + / - standard error of the mean (SEM). [Figure 6] Figure 6A is a graph showing an experiment performed in a mouse model of non-small cell lung cancer using humanized 5D8 antibody, and Figure 6B is a graph showing an experiment performed in a mouse model of non-small cell lung cancer using r5D8 antibody. [Figure 7]Figure 7A shows the effect of h5D8 on the inhibition of U251 cell growth in an orthotopic mouse model of GBM. Quantification at day 26 is shown. Figure 7B shows data from mice inoculated with luciferase-expressing human U251 GBM cells and subsequently treated twice weekly with 100, 200, or 300 μg of h5D8 or vehicle. Tumor size was determined by bioluminescence (Xenogen IVIS Spectrum) on day 7. The graph shows individual tumor measurements with horizontal bars indicating the mean ± SEM. Statistical significance was calculated using the unpaired, nonparametric Mann-Whitney U test. [Figure 8] Figure 8A shows the effect of r5D8 on inhibiting ovarian cancer cell growth in a syngeneic mouse model. Figure 8B shows individual tumor measurements at day 25. Figure 8C illustrates that h5D8 significantly reduced tumor growth (p<0.05) when administered at 200 μg / mouse twice weekly. Symbols are mean ± SEM, and statistical significance was compared to vehicle (using an unpaired, nonparametric Mann-Whitney U test). [Figure 9] Figure 9A shows the effect of r5D8 on inhibiting the growth of colorectal cancer cells in a syngeneic mouse model, and Figure 9B shows individual tumor measurements at day 17. [Figure 10-1]Figure 10A shows reduced macrophage infiltration into tumor sites in an orthotopic mouse model of GBM, with representative images and quantification of CCL22+ cells. Figure 10B shows reduced macrophage infiltration in a human organotypic explant culture model. Figure 10C shows reduced macrophage infiltration into tumor sites in a syngeneic mouse model of ovarian cancer, with representative images and quantification of CCL22+ cells. Figure 10D shows reduced macrophage infiltration into tumor sites in a syngeneic mouse model of colorectal cancer, with representative images and quantification of CCL22+ cells. Figure 10E shows the inflammatory phenotype of tumor-associated macrophages (TAMs) harvested from tumors treated with h5D8 (15 mg / kg, 2QW) on day 25 (endpoint). TAMs in treated tumors were polarized to the M1 pro-inflammatory phenotype. Statistical significance was determined by unpaired t-test. FIG. 10F shows gene expression data for monocytes cultured with conditioned medium from LIF knockdown cells. [Figure 10-2] Same as above. [Figure 10-3] Same as above. [Figure 10-4] Same as above. [Figure 10-5] Same as above. [Figure 11-1] Figure 11A shows an increase in non-myeloid effector cells in a syngeneic mouse model of ovarian cancer after treatment with r5D8. Figure 11B shows an increase in non-myeloid effector cells in a syngeneic mouse model of colorectal cancer after treatment with r5D8. Figure 11C shows a decrease in the percentage of CD4+ Treg cells in a mouse model of NSCLC cancer after treatment with r5D8. [Figure 11-2] Same as above. [Figure 11-3] Same as above. [Figure 12]Figure 12 shows data from CT26 tumor-bearing mice treated twice weekly with PBS (control) or r5D8 administered intraperitoneally in the presence or absence of anti-CD4 and anti-CD8 depleting antibodies. The graph shows individual tumor measurements at day 13, expressed as mean tumor volume + SEM. Statistical significance between groups was determined by unpaired, nonparametric Mann-Whitney U test. R5D8 inhibited CT26 tumor growth (*p<0.05). Tumor growth inhibition by r5D8 was significantly reduced in the presence of anti-CD4 and anti-CD8 depleting antibodies (****p<0.0001). [Figure 13] Figure 13A illustrates the overall co-crystal structure of h5D8 Fab in complex with LIF. The location of the gp130 interaction site is indicated on the surface of LIF (highlighted). Figure 13B illustrates the detailed interactions between LIF and h5D8, showing residues that form salt bridges and h5D8 residues with a buried surface area of ​​over 100 Å. [Figure 14] Figure 14A illustrates a superposition of five h5D8 Fab crystal structures, demonstrating a high degree of similarity despite being crystallized under different chemical conditions. Figure 14B illustrates the extensive network of van der Waals interactions mediated by unpaired Cys100. This residue is well-ordered and participates in shaping the HCDR1 and HCDR3 conformations, without participating in undesired disulfide scrambling. Distances between residues are shown as dashed lines and labeled. [Figure 15] Figure 15A illustrates the binding of h5D8 C100 mutants to human LIF by ELISA. Figure 15B illustrates the binding of h5D8 C100 mutants to mouse LIF by ELISA. [Figure 16]Figure 16A shows that h5D8 does not block the binding between LIF and LIFR by Octet. Binding of h5D8 to LIF is followed by subsequent binding to LIFR. Figures 16B-16C illustrate ELISA analysis of LIF / mAb complex binding to immobilized LIFR or gp130. Signals from species-specific peroxidase-conjugated anti-IgG antibodies (anti-human for (-) and h5D8, anti-rat for r5d8 and B09) detect the antibody moiety of the mAb / LIF complex that binds to immobilized LIFR (Figure 16B) or gp130 (Figure 16C) coated plates. [Figure 17-1] 17A-17B illustrate the mRNA expression of LIF (FIG. 17A) or LIFR (FIG. 18B) in 72 different human tissues. [Figure 17-2] Same as above. [Figure 18] Figure 18 shows data from mice implanted with subcutaneous CT26 tumors treated with h5D8 and cisplatin. The figure shows the time course of tumor growth by mean tumor volume for vehicle, control IgG1, h5D8 alone, cisplatin alone, and the h5D8 + cisplatin combination groups. [Figure 19] Figures 19A-19B show tumor volume data over time from mice implanted subcutaneously with CT26 tumors and treated with h5D8 and doxorubicin (Figure 19A) or h5D8 and paclitaxel (Figure 19B). [Figure 20] Figure 20 shows data from mice treated with h5D8 and cisplatin and implanted intradermally with CT26 tumors. The figure shows the time course of tumor growth by mean tumor volume for vehicle, control IgG1, h5D8 alone, cisplatin alone, and the h5D8 + cisplatin combination groups. DETAILED DESCRIPTION OF THE INVENTION

[0014] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Any reference to "or" herein is intended to include "and / or" unless specified otherwise.

[0015] In one aspect, described herein is the use of a leukemia inhibitory factor (LIF) binding polypeptide in combination with a platinum-based anti-neoplastic agent to treat cancer in an individual.

[0016] In another aspect, described herein is the use of a leukemia inhibitory factor (LIF)-binding antibody in combination with a platinum-based anti-cancer agent to treat cancer in an individual, wherein the LIF-binding antibody comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOS: 1-3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOS: 4 or 5; or (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR4) comprising the amino acid sequence set forth in any one of SEQ ID NOS: 6-8. (d) immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13.

[0017] In another aspect, described herein is a method of treating an individual with cancer, comprising administering to the individual with cancer effective amounts of: (a) a leukemia inhibitory factor (LIF)-binding polypeptide; and (b) a platinum-based anti-cancer agent.

[0018] In another aspect, described herein is a method of treating an individual with cancer, the method comprising administering an effective amount of a leukemia inhibitory factor (LIF)-binding polypeptide to an individual with cancer who is receiving a therapeutic amount of a platinum-based anti-cancer agent.

[0019] In another aspect, described herein is a method of treating an individual with cancer, the method comprising administering an effective amount of a platinum-based anti-neoplastic agent to an individual with cancer who has been administered a therapeutic amount of a leukemia inhibitory factor (LIF)-binding polypeptide.

[0020] In another aspect, described herein is a method of treating an individual with cancer, comprising administering to an individual an effective amount of: (a) (i) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1-3; (ii) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (iii) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6-8; (iv) The method includes administering to an individual having cancer a leukemia inhibitory factor (LIF)-binding antibody comprising: (a) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (b) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (c) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13; and (c) a platinum-based anti-cancer agent.

[0021] In another aspect, described herein is a kit that includes: (a) a leukemia inhibitory factor (LIF) binding polypeptide; and (b) a platinum-based anti-neoplastic agent.

[0022] In another aspect, described herein is a composition comprising: (a) a leukemia inhibitory factor (LIF)-binding polypeptide; and (b) a platinum-based anti-neoplastic agent.

[0023] As used herein, the terms "individual," "subject," and "patient" are used interchangeably and include a human who has been diagnosed with or is suspected of having a tumor, cancer, or other neoplasm.

[0024] The terms "cancer" and "tumor" refer to a physiological condition in mammals characterized by unregulated cell growth. Cancer is a class of diseases in which a group of cells exhibit uncontrolled or unwanted growth. Cancer cells can also spread to other locations, which can result in the formation of metastases. The spread of cancer cells within the body can occur, for example, via lymph or blood. Uncontrolled growth, invasion, and metastasis formation are also referred to as malignant characteristics of cancer. These malignant characteristics distinguish cancer from benign tumors, which generally do not invade or metastasize.

[0025] As used herein, the term "effective amount" refers to the amount of a therapeutic agent that produces a biological effect when administered to a mammal. Biological effects include, but are not limited to, inhibiting or blocking receptor-ligand interactions (e.g., LIF-LIFR), inhibiting signal transduction pathways (e.g., STAT3 phosphorylation), reducing tumor growth, reducing tumor metastasis, or prolonging survival of tumor-bearing animals. A "therapeutic amount" is the concentration of a drug calculated to produce a therapeutic effect. A therapeutic amount encompasses a range of dosages capable of eliciting a therapeutic response in a population of individuals. The mammal may be a human individual. The human individual may be suffering from or suspected of suffering from a tumor.

[0026] As used herein, unless otherwise indicated, the term "antibody" includes antigen-binding fragments of antibodies, i.e., antibody fragments that retain the ability to specifically bind to the antigen bound by the full-length antibody, e.g., fragments that retain one or more CDR regions. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab'), and Fv fragments; bispecific antibodies; linear antibodies; heavy chain antibodies; single-chain antibody molecules, e.g., single-chain variable region fragments (scFvs), nanobodies, and multispecific antibodies formed from antibody fragments with different specificities, such as bispecific antibodies. In certain embodiments, antibodies are humanized to reduce an individual's immune response to the antibody. For example, antibodies can be chimeric, e.g., non-human variable regions with human constant regions, or CDR-grafted, e.g., non-human CDR regions with human constant and variable region framework sequences. In certain embodiments, antibodies are deimmunized after humanization. Deimmunization involves removing or mutating one or more T-cell epitopes within the constant region of the antibody. In certain embodiments, the antibodies described herein are monoclonal. As used herein, a "recombinant antibody" is an antibody that contains amino acid sequences from two different species or two different sources, and includes synthetic molecules, e.g., antibodies that contain non-human CDRs and human framework or constant regions. In certain embodiments, recombinant antibodies of the invention are produced from recombinant DNA molecules or are synthesized.

[0027] The percent (%) of sequence identity with respect to a reference polypeptide or antibody sequence is the percentage of amino acid residues in a candidate sequence that are identical to those in the reference polypeptide or antibody sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent of sequence identity, and without considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in a variety of known ways, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. It is possible to determine appropriate parameters for aligning sequences, including the algorithm required to achieve maximum alignment across the entire length of the sequences being compared. However, for purposes of this specification, percent amino acid sequence identity values ​​are generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was created by Genentech, Inc., and the source code, along with user documentation, is deposited in the U.S. Copyright Office, Washington, DC, 20559, where it is registered under U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, Calif., or can be compiled from the source code. The ALIGN-2 program should be compiled for use on UNIX operating systems, including digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary.

[0028] When ALIGN-2 is used for amino acid sequence comparison, the percent amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B (which can alternatively be expressed as a given amino acid sequence A having or containing a certain percent amino acid sequence identity to, with, or against a given amino acid sequence B) is calculated as follows: 100 × fraction X / Y (where X is the number of amino acid residues scored as identical matches by the sequence alignment program ALIGN-2 for the alignment of A and B, and Y is the total number of amino acid residues in B). It should be understood that if the length of amino acid sequence A is not equal to the length of amino acid sequence B, the percent amino acid sequence identity of A to B will not be equal to the percent amino acid sequence identity of B to A. Unless otherwise specifically specified, all percent amino acid sequence identity values ​​used herein are obtained as described in the immediately preceding paragraph using the ALIGN-2 computer program.

[0029] The term "epitope" includes any determinant capable of being bound by an antigen-binding protein such as an antibody. An epitope is a region of an antigen that is bound by an antigen-binding protein that targets that antigen, and, if the antigen is a protein, includes specific amino acids that make direct contact with the antigen-binding protein. Epitopes are most often found on proteins but can also be found on other types of molecules, such as sugars or lipids. Epitopes can include chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl, or sulfonyl groups, and can have specific three-dimensional structural characteristics and / or specific charge characteristics. In general, an antibody specific for a particular target antigen will preferentially recognize an epitope on the target antigen in a complex mixture of proteins and / or macromolecules.

[0030] The term "antineoplastic agent" refers to an agent capable of acting to prevent, inhibit, or stop the development of cancer, tumors, or neoplasms. Antineoplastic agents may also be known as chemotherapeutic agents. Antineoplastic agents can target and destroy cancer cells. Antineoplastic agents may include, but are not limited to, antimetabolites, biological response modifiers, bleomycin, DNA alkylating agents, DNA cross-linking agents, enzymes, hormones, monoclonal antibodies, platinum complexes, proteasome inhibitors, taxanes, vincas, topoisomerase inhibitors, tyrosine kinase inhibitors, nucleoside analogs, antifolates, anthracyclines, podophyllotoxins, alkylating agents, mTOR inhibitors, retinoids, histone deacetylase inhibitors, and immunomodulators.

[0031] Structural Features of the Antibodies Described Herein Complementarity-determining regions ("CDRs") are parts of immunoglobulin (antibody) variable regions that are primarily responsible for the antigen-binding specificity of the antibody. CDR regions are highly variable from antibody to antibody, even when the antibodies specifically bind to the same target or epitope. The heavy chain variable region contains three CDR regions, abbreviated as VH-CDR1, VH-CDR2, and VH-CDR3; the light chain variable region contains three CDR regions, abbreviated as VL-CDR1, VL-CDR2, and VL-CDR3. These CDR regions are arranged contiguously within the variable region, with CDR1 being the most N-terminal and CDR3 being the most C-terminal. The CDRs are interposed between framework regions that contribute to the structure and exhibit much less variability than the CDR regions. The heavy chain variable region comprises four framework regions, abbreviated as VH-FR1, VH-FR2, VH-FR3, and VH-FR4; the light chain variable region comprises four framework regions, abbreviated as VL-FR1, VL-FR2, VL-FR3, and VL-FR4. A complete, full-size, bivalent antibody comprising two heavy and light chains will comprise 12 CDRs, including three unique heavy chain CDRs and three unique light chain CDRs; and 16 FR regions, including four unique heavy chain FR regions and four unique light chain FR regions. In certain embodiments, the antibodies described herein comprise a minimum of three heavy chain CDRs. In certain embodiments, the antibodies described herein comprise a minimum of three light chain CDRs. In certain embodiments, the antibodies described herein comprise a minimum of three heavy chain CDRs and three light chain CDRs.The exact amino acid sequence boundaries of a given CDR or FR can be determined using the Kabat numbering scheme described in Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme); Al-Lazikani et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme); MacCallum et al., J. Mol. Biol. 262:732-745 (1996), "Antibody-antigen interactions: Contact analysis and binding site topography," ("Contact" numbering scheme); Lefranc MP et al., "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains," Dev Comp Immunol, 2003 Jan;27(1):55-77 ("IMGT" numbering scheme); and Honegger A and Plueckthun A, "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool," J Mol Biol, 2001 Jun 8;309(3):657-70, ("Aho" numbering scheme). CDRs are identified herein from the variable sequences provided herein using different numbering systems, including the Kabat, IMGT, Chothia numbering systems, or any combination of the three. The boundaries of a given CDR or FR may vary depending on the scheme used for identification.For example, the Kabat scheme is based on structural alignment, while the Chothia scheme is based on structural information. The numbering for both the Kabat and Chothia schemes is based on the most common antibody region sequence lengths, along with insertions (addressed by an insertion letter, e.g., "30a") and deletions that appear in some antibodies. The two schemes place certain insertions and deletions ("indels") in different positions, resulting in different numbering. The Contact scheme is based on analysis of complex crystal structures and is similar in many ways to the Chothia numbering scheme.

[0032] The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to an antigen. The variable domains of the heavy and light chains (VH and VL, respectively) of natural antibodies generally have similar structures, with each domain containing four conserved framework regions (FR) and three CDRs (see, for example, Kindt et al., Kuby Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007)). Only one VH or VL domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind to a specific antigen can be isolated using a VH or VL domain from an antigen-binding antibody to screen a library of complementary VL or VH domains, respectively (see, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991)). In certain embodiments, the antibodies described herein are humanized. In certain embodiments, the antibodies described herein are chimeric. In certain embodiments, the antibodies described herein comprise variable regions of rat origin. In certain embodiments, the antibodies described herein comprise CDRs of rat origin. In certain embodiments, the antibodies described herein comprise variable regions of murine origin. In certain embodiments, the antibodies described herein comprise CDRs of murine origin.

[0033] For example, to improve antibody affinity, alterations (e.g., substitutions) can be made to CDRs. Such alterations can be made to CDR-encoding codons with high mutation rates during somatic maturation (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and the resulting variants can be tested for binding affinity. Affinity maturation (e.g., using error-prone PCR, chain shuffling, CDR randomization, or oligonucleotide-directed mutagenesis) can be used to improve antibody affinity (see, e.g., Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (2001)). CDR residues involved in antigen binding can be specifically identified, for example, using alanine scanning mutagenesis or modeling (see, e.g., Cunningham and Wells Science, 244:1081-1085 (1989)). Specifically, CDR-H3 and CDR-L3 are often targeted. Alternatively, or in addition, the crystal structure of the antigen-antibody complex is analyzed to identify contact points between the antibody and the antigen. These contact residues and adjacent residues can be targeted or eliminated as candidates for substitution. Variants can be screened to determine whether they contain the desired properties.

[0034] In certain embodiments, the antibodies described herein comprise a constant region in addition to the variable region. H ) is located at the C-terminus of the complete heavy chain polypeptide, C-terminal to the variable region, H 1. C H 2. C H 3, and C H It contains four domains, abbreviated as C4. The light chain constant region (C L ) is C Hand is located at the C-terminus of the complete light chain polypeptide, C-terminal to the variable region. The constant region is highly conserved, with different isotypes associated with slightly different functions and properties. In certain embodiments, the constant region is unnecessary for antibody binding to a target antigen. In certain embodiments, the antibody constant regions, both heavy and light, are unnecessary for antibody binding. In certain embodiments, the antibodies described herein lack one or more light chain constant regions, heavy chain constant regions, or both. Most monoclonal antibodies are of the IgG isotype, which is further divided into four subclasses: IgG1, IgG2, IgG3, and IgG4. In certain embodiments, the antibodies described herein comprise any IgG subclass. In certain embodiments, the IgG subclass comprises IgG1. In certain embodiments, the IgG subclass comprises IgG2. In certain embodiments, the IgG subclass comprises IgG3. In certain embodiments, the IgG subclass comprises IgG4.

[0035] Antibodies contain a fragment crystallizable region (Fc region) that is responsible for binding to complement and Fc receptors. The Fc region is the C region of the antibody molecule. H 2. C H 3, and C HThe Fc region of an antibody comprises four regions. The Fc region of an antibody is responsible for activating complement and antibody-dependent cellular cytotoxicity (ADCC). The Fc region also contributes to the serum half-life of the antibody. In certain embodiments, the Fc region of an antibody described herein comprises one or more amino acid substitutions that promote complement-mediated cytolysis. In certain embodiments, the Fc region of an antibody described herein comprises one or more amino acid substitutions that promote ADCC. In certain embodiments, the Fc region of an antibody described herein comprises one or more amino acid substitutions that decrease complement-mediated cytolysis. In certain embodiments, the Fc region of an antibody described herein comprises one or more amino acid substitutions that increase binding of the antibody to an Fc receptor. In certain embodiments, the Fc receptor comprises FcγRI (CD64), FcγRIIA (CD32), FcγRIIIA (CD16a), FcγRIIIB (CD16b), or any combination thereof. In certain embodiments, the Fc region of an antibody described herein comprises one or more amino acid substitutions that increase the serum half-life of the antibody. In certain embodiments, the one or more amino acid substitutions that increase the serum half-life of the antibody increase the affinity of the antibody for the neonatal Fc receptor (FcRn).

[0036] In some embodiments, the antibodies of this disclosure are variants that retain some, but not all, effector functions, making them desirable candidates for applications in which in vivo antibody half-life is important but certain effector functions (such as complement and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity tests can be performed to confirm reduced / loss of CDC and / or ADCC activity. For example, Fc receptor (FcR) binding assays can be performed to ensure that the antibody lacks FcγR binding (and thus likely lacks ADCC activity) but retains FcRn binding ability. Non-limiting examples of in vitro assays for assessing ADCC activity of molecules of interest are described in U.S. Patent Nos. 5,500,362 and 5,821,337. Alternatively, non-radioactive assay methods (e.g., ACTI™ and CytoTox 96® non-radioactive cytotoxicity tests) can be used. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC), monocytes, macrophages, and natural killer (NK) cells.

[0037] The antibodies can have increased half-life and increased binding to neonatal Fc receptor (FcRn) (see, e.g., U.S. Patent Application Publication No. 2005 / 0014934). Such antibodies can comprise an Fc region having one or more substitutions therein that improve binding of the Fc region to FcRn, including those having a substitution at one or more of the following Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, ​​413, 424, or 434 (according to the EU numbering system) (see, e.g., U.S. Patent No. 7,371,826). Other examples of Fc region variants are also contemplated (see, e.g., Duncan & Winter, Nature 322:738-40 (1988); U.S. Patent Nos. 5,648,260 and 5,624,821; and WO 94 / 29351).

[0038] Antibodies useful in clinical practice are often "humanized" to reduce immunogenicity in human individuals. Humanized antibodies improve the safety and efficacy of monoclonal antibody therapy. One common humanization method is to produce a monoclonal antibody in any suitable animal (e.g., mouse, rat, hamster) and replace the constant region with a human constant region; antibodies engineered in this manner are called "chimeras." Another common method is "CDR grafting," in which non-human V-FRs are replaced with human V-FRs. In the CDR grafting method, all residues except the CDR regions are of human origin. In certain embodiments, the antibodies described herein are humanized. In certain embodiments, the antibodies described herein are chimeric. In certain embodiments, the antibodies described herein are CDR-grafted.

[0039] Humanization generally reduces the overall affinity of an antibody or has little effect on the overall affinity of an antibody. Described herein are antibodies that unexpectedly have greater affinity for their target after humanization. In certain embodiments, humanization increases affinity for the antibody by 10%. In certain embodiments, humanization increases affinity for the antibody by 25%. In certain embodiments, humanization increases affinity for the antibody by 35%. In certain embodiments, humanization increases affinity for the antibody by 50%. In certain embodiments, humanization increases affinity for the antibody by 60%. In certain embodiments, humanization increases affinity for the antibody by 75%. In certain embodiments, humanization increases affinity for the antibody by 100%. Affinity is suitably measured using surface plasmon resonance (SPR). In certain embodiments, affinity is measured using glycosylated human LIF. In certain embodiments, glycosylated human LIF is immobilized on the surface of an SPR chip. In certain embodiments, the antibody has a K of less than about 300 nanomolar, 200 nanomolar, 150 nanomolar, 125 nanomolar, 100 nanomolar, 90 nanomolar, 80 nanomolar, 70 nanomolar, 60 nanomolar, 50 nanomolar, 40 nanomolar, or less. D Combine with.

[0040] The compositions and methods described herein include a combination of a LIF-binding polypeptide and a platinum-based anti-neoplastic agent. In certain embodiments, the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. In certain embodiments, the LIF-binding polypeptide is an antibody that specifically binds to LIF. In certain embodiments, the LIF-binding antibody comprises at least one framework region derived from a human immunoglobulin framework region. In certain embodiments, the antibody that specifically binds to LIF is humanized. In certain embodiments, the antibody that specifically binds to LIF is deimmunized. In certain embodiments, the antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. In certain embodiments, the antibody that specifically binds to LIF is an IgG antibody. In certain embodiments, the antibody that specifically binds to LIF is a Fab, F(ab)2, single-domain antibody, single-chain variable fragment (scFv), or nanobody. In certain embodiments, the antibody is the h5D8 antibody described herein.

[0041] In certain embodiments described herein, the antibody used or administered in combination with a platinum-based anti-cancer agent is the h5D8 antibody. The h5d8 antibody specifically binds to LIF and comprises a VH-CDR1 set forth in any one of SEQ ID NOS: 1 to 3, a VH-CDR2 set forth in any one of SEQ ID NOS: 4 or 5, and a VH-CDR3 set forth in any one of SEQ ID NOS: 6 to 8. In certain embodiments described herein, h5D8 specifically binds to LIF and comprises a VL-CDR1 set forth in any one of SEQ ID NOS: 9 or 10, a VL-CDR2 set forth in SEQ ID NOS: 11 or 12, and a VL-CDR3 set forth in SEQ ID NOS: 13. In certain embodiments described herein, h5D8 specifically binds to LIF and comprises a VH-CDR1 set forth in any one of SEQ ID NOS: 1-3, a VH-CDR2 set forth in any one of SEQ ID NOS: 4 or 5, a VH-CDR3 set forth in any one of SEQ ID NOS: 6-8, a VL-CDR1 set forth in any one of SEQ ID NOS: 9 or 10, a VL-CDR2 set forth in SEQ ID NOS: 11 or 12, and a VL-CDR3 set forth in SEQ ID NOS: 13. The VH and VL regions of h5D8 are set forth in SEQ ID NOS: 42 and 46.

[0042] In certain embodiments, an antibody that specifically binds to LIF comprises one or more human heavy chain framework regions comprising a VH-FR1 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 14-17; a VH-FR2 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 18 or 19; a VH-FR3 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 20-22; or a VH-FR4 region amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 23-25. In certain embodiments, the one or more human heavy chain framework regions comprise a VH-FR1 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 20, and a VH-FR4 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 24. In certain embodiments, an antibody that specifically binds to LIF comprises one or more human light chain framework regions comprising a VL-FR1 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 26-29; a VL-FR2 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 30-33; a VL-FR3 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 34-37; or a VL-FR4 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 38-40.In certain embodiments, the one or more human light chain framework regions comprise a VL-FR1 amino acid sequence that is at least about 80%, about 90%, or about 95% identical to the amino acid sequence set forth in SEQ ID NO:27, a VL-FR2 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:31, a VL-FR3 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:35, and a VL-FR4 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:38. In certain embodiments, the one or more human heavy chain framework regions and the one or more human light chain regions comprise a VH-FR1 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 15; a VH-FR2 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 19; a VH-FR3 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 20; comprises a VH-FR4 amino acid sequence that is 99% identical, a VL-FR1 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:27, a VL-FR2 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:31, a VL-FR3 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:35, and a VL-FR4 amino acid sequence that is at least 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:38.In certain embodiments, an antibody that specifically binds to LIF comprises one or more human heavy chain framework regions comprising a VH-FR1 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 14 to 17, a VH-FR2 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 18 or 19, a VH-FR3 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 20 to 22, or a VH-FR4 region amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 23 to 25. In certain embodiments, the one or more human heavy chain framework regions comprise a VH-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 20, and a VH-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 24. In certain embodiments, an antibody that specifically binds to LIF comprises one or more human light chain framework regions comprising a VL-FR1 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 26 to 29, a VL-FR2 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 30 to 33, a VL-FR3 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 34 to 37, or a VL-FR4 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 38 to 40. In certain embodiments, the one or more human light chain framework regions comprise a VL-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 27, a VL-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 31, a VL-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 35, and a VL-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 38.In certain embodiments, the one or more human heavy chain framework regions and the one or more human light chain regions comprise a VH-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 20, a VH-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 24, a VL-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 27, a VL-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 31, a VL-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 35, and a VL-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 38. In certain embodiments, the antibody specifically binds to human LIF.

[0043] The r5D8 antibody described herein was produced from rats immunized with DNA encoding human LIF. r5D8 was cloned and sequenced. It contains CDRs (using a combination of the Kabat and IMGT CDR numbering systems) that include the following amino acid sequences: VH-CDR1 corresponding to SEQ ID NO: 1 (GFTFSHAWMH), VH-CDR2 corresponding to SEQ ID NO: 4 (QIKAKSDDYATYYAESVKG), VH-CDR3 corresponding to SEQ ID NO: 6 (TCWEWDLDF), VL-CDR1 corresponding to SEQ ID NO: 9 (RSSQSLLDSDGHTYLN), VL-CDR2 corresponding to SEQ ID NO: 11 (SVSNLES), and VL-CDR3 corresponding to SEQ ID NO: 13 (MQATHAPPYT). This antibody was humanized by CDR grafting, and the humanized version is referred to as h5D8.

[0044] In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a VH-CDR1 at least 80% or 90% identical to that set forth in SEQ ID NO:1 (GFTFSHAWMH), a VH-CDR2 at least 80%, 90%, or 95% identical to that set forth in SEQ ID NO:4 (QIKAKSDDYATYYAESVKG), and a VH-CDR3 at least 80% or 90% identical to that set forth in SEQ ID NO:6 (TCWEWDLDF). In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a VL-CDR1 at least 80% or 90% identical to that set forth in SEQ ID NO:9 (RSSQSLLDSDGHTYLN), a VL-CDR2 at least 80% identical to that set forth in SEQ ID NO:11 (SVSNLES), and a VL-CDR3 at least 80% or 90% identical to that set forth in SEQ ID NO:13 (MQATHAPPYT). In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a VH-CDR1 set forth in SEQ ID NO: 1 (GFTFSHAWMH), a VH-CDR2 set forth in SEQ ID NO: 4 (QIKAKSDDYATYYAESVKG), a VH-CDR3 set forth in SEQ ID NO: 6 (TCWEWDLDF), a VL-CDR1 set forth in SEQ ID NO: 9 (RSSQSLLDSDGHTYLN), a VL-CDR2 set forth in SEQ ID NO: 11 (SVSNLES), and a VL-CDR3 set forth in SEQ ID NO: 13 (MQATHAPPYT). Some conservative amino acid substitutions in the amino acid sequences of the CDRs of this disclosure are contemplated. In certain embodiments, the antibody comprises a CDR that differs by 1, 2, 3, or 4 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 6, 9, 11, and 13. In certain embodiments, the antibody comprises a CDR that differs by 1, 2, 3, or 4 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 6, 9, 11, and 13 and does not affect binding affinity by more than 10%, 20%, or 30%.In certain embodiments, an antibody that specifically binds to LIF comprises one or more human heavy chain framework regions comprising a VH-FR1 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 14-17, a VH-FR2 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 18 or 19, a VH-FR3 amino acid sequence that is at least 90% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 20-22, or a VH-FR4 region amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 23-25. In certain embodiments, the one or more human heavy chain framework regions comprise a VH-FR1 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 20, and a VH-FR4 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 24. In certain embodiments, an antibody that specifically binds to LIF comprises one or more human light chain framework regions comprising a VL-FR1 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 26-29; a VL-FR2 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 30-33; a VL-FR3 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 34-37; or a VL-FR4 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 38-40.In certain embodiments, the one or more human light chain framework regions comprise a VL-FR1 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:27, a VL-FR2 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:31, a VL-FR3 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:35, and a VL-FR4 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:38. In certain embodiments, the one or more human heavy chain framework regions and the one or more human light chain regions comprise a VH-FR1 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 20, a VH-FR4 amino acid sequence at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 24, or a VH-FR5 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 25. a VL-FR1 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 27, a VL-FR2 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 31, a VL-FR3 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 35, and a VL-FR4 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 38. In certain embodiments, the antibody specifically binds to human LIF.In certain embodiments, an antibody that specifically binds to LIF comprises one or more human heavy chain framework regions comprising a VH-FR1 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 14 to 17, a VH-FR2 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 18 or 19, a VH-FR3 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 20 to 22, or a VH-FR4 region amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 23 to 25. In certain embodiments, the one or more human heavy chain framework regions comprise a VH-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 20, and a VH-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 24. In certain embodiments, an antibody that specifically binds to LIF comprises one or more human light chain framework regions comprising a VL-FR1 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 26 to 29, a VL-FR2 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 30 to 33, a VL-FR3 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 34 to 37, or a VL-FR4 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 38 to 40. In certain embodiments, the one or more human light chain framework regions comprise a VL-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 27, a VL-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 31, a VL-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 35, and a VL-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 38.In certain embodiments, the one or more human heavy chain framework regions and the one or more human light chain regions comprise a VH-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 20, a VH-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 24, a VL-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 27, a VL-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 31, a VL-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 35, and a VL-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 38. In certain embodiments, the antibody specifically binds to human LIF.

[0045] In certain embodiments, described herein are antibodies that specifically bind to LIF comprising a VH-CDR1 amino acid sequence at least 80% or 90% identical to that set forth in SEQ ID NO:1 (GFTFSHAWMH), a VH-CDR2 amino acid sequence at least 80%, 90%, or 95% identical to that set forth in SEQ ID NO:4 (QIKAKSDDYATYYAESVKG), and a VH-CDR3 amino acid sequence at least 80% or 90% identical to that set forth in SEQ ID NO:8 (TSWEWDLDF). In certain embodiments, described herein are antibodies that specifically bind to LIF comprising a VL-CDR1 amino acid sequence at least 80% or 90% identical to that set forth in SEQ ID NO:9 (RSSQSLLDSDGHTYLN), a VL-CDR2 amino acid sequence at least 80% identical to that set forth in SEQ ID NO:11 (SVSNLES), and a VL-CDR3 amino acid sequence at least 80% or 90% identical to that set forth in SEQ ID NO:13 (MQATHAPPYT). In certain embodiments, described herein are antibodies that specifically bind to LIF comprising the VH-CDR1 amino acid sequence set forth in SEQ ID NO: 1 (GFTFSHAWMH), the VH-CDR2 amino acid sequence set forth in SEQ ID NO: 4 (QIKAKSDDYATYYAESVKG), the VH-CDR3 amino acid sequence set forth in SEQ ID NO: 8 (TSWEWDLDF), the VL-CDR1 amino acid sequence set forth in SEQ ID NO: 9 (RSSQSLLDSDGHTYLN), the VL-CDR2 amino acid sequence set forth in SEQ ID NO: 11 (SVSNLES), and the VL-CDR3 amino acid sequence set forth in SEQ ID NO: 13 (MQATHAPPYT). Some conservative amino acid substitutions in the amino acid sequences of the CDRs of this disclosure are contemplated. In certain embodiments, the antibody comprises a CDR that differs by 1, 2, 3, or 4 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 8, 9, 11, and 13. In certain embodiments, the antibody comprises a CDR that differs by 1, 2, 3, or 4 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 8, 9, 11, and 13 and does not affect binding affinity by more than 10%, 20%, or 30%.In certain embodiments, an antibody that specifically binds to LIF comprises one or more human heavy chain framework regions comprising a VH-FR1 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 14-17, a VH-FR2 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 18 or 19, a VH-FR3 amino acid sequence that is at least 90% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 20-22, or a VH-FR4 region amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 23-25. In certain embodiments, the one or more human heavy chain framework regions comprise a VH-FR1 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 20, and a VH-FR4 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 24. In certain embodiments, an antibody that specifically binds to LIF comprises one or more human light chain framework regions comprising a VL-FR1 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 26-29; a VL-FR2 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 30-33; a VL-FR3 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 34-37; or a VL-FR4 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 38-40.In certain embodiments, the one or more human light chain framework regions comprise a VL-FR1 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:27, a VL-FR2 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:31, a VL-FR3 amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:35, and a VL-FR4 amino acid sequence that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO:38. In certain embodiments, the one or more human heavy chain framework regions and the one or more human light chain regions comprise a VH-FR1 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 20, a VH-FR4 amino acid sequence at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 24, or a VH-FR5 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 25. a VL-FR1 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 27, a VL-FR2 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 31, a VL-FR3 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 35, and a VL-FR4 amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 38. In certain embodiments, the antibody specifically binds to human LIF.In certain embodiments, an antibody that specifically binds to LIF comprises one or more human heavy chain framework regions comprising a VH-FR1 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 14 to 17, a VH-FR2 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 18 or 19, a VH-FR3 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 20 to 22, or a VH-FR4 region amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 23 to 25. In certain embodiments, the one or more human heavy chain framework regions comprise a VH-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 20, and a VH-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 24. In certain embodiments, an antibody that specifically binds to LIF comprises one or more human light chain framework regions comprising a VL-FR1 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 26 to 29, a VL-FR2 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 30 to 33, a VL-FR3 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 34 to 37, or a VL-FR4 amino acid sequence identical to the amino acid sequence set forth in any one of SEQ ID NOs: 38 to 40. In certain embodiments, the one or more human light chain framework regions comprise a VL-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 27, a VL-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 31, a VL-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 35, and a VL-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 38.In certain embodiments, the one or more human heavy chain framework regions and the one or more human light chain regions comprise a VH-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 15, a VH-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 19, a VH-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 20, a VH-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 24, a VL-FR1 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 27, a VL-FR2 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 31, a VL-FR3 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 35, and a VL-FR4 amino acid sequence identical to the amino acid sequence set forth in SEQ ID NO: 38. In certain embodiments, the antibody specifically binds to human LIF.

[0046] In certain embodiments, described herein are antibodies that specifically bind to LIF comprising a humanized heavy chain variable region comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, and 44. In certain embodiments, described herein are antibodies that specifically bind to LIF comprising a humanized heavy chain variable region comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, described herein are antibodies that specifically bind to LIF comprising a humanized light chain variable region comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, the antibodies specifically bind to human LIF.

[0047] In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain variable region comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 42; and a humanized light chain variable region comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 42; and a humanized light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 46.

[0048] In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain variable region comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 66; and a humanized light chain variable region comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 66; and a humanized light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 46.

[0049] In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60; and a humanized light chain comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain comprising an amino acid sequence set forth in any one of SEQ ID NOs: 57-60; and a humanized light chain comprising the amino acid sequence set forth in any one of SEQ ID NOs: 61-64.

[0050] In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 58; and a humanized light chain comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 62. In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 58; and a humanized light chain comprising the amino acid sequence set forth in SEQ ID NO: 62. In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 67; and a humanized light chain comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 62. In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 67; and a humanized light chain comprising the amino acid sequence set forth in SEQ ID NO: 62.

[0051] In certain embodiments, described herein is a recombinant antibody that specifically binds to leukemia inhibitory factor (LIF), comprising a heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:3; a heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:4; a heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:7; a light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:9; and a light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:11; and a light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:13.

[0052] In certain embodiments, described herein is a recombinant antibody that specifically binds to leukemia inhibitory factor (LIF), comprising: a heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:2; a heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:5; a heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:6; a light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:10; and a light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:12; and a light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:13. Some conservative amino acid substitutions in the amino acid sequences of the CDRs of this disclosure are contemplated. In certain embodiments, the antibody comprises a CDR that differs by 1, 2, 3, or 4 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs:2, 5, 6, 10, 12, and 13. In certain embodiments, the antibody comprises a CDR that differs by 1, 2, 3, or 4 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs: 2, 5, 6, 10, 12, and 13 and does not affect binding affinity by more than 10%, 20%, or 30%.

[0053] In certain embodiments, described herein is a recombinant antibody that specifically binds to leukemia inhibitory factor (LIF), comprising: a heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:3; a heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:4; a heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:7; a light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:9; and a light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:11; and a light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:13. Some conservative amino acid substitutions in the amino acid sequences of the CDRs of this disclosure are contemplated. In certain embodiments, the antibody comprises a CDR that differs by 1, 2, 3, or 4 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs:3, 4, 7, 9, 11, and 13. In certain embodiments, the antibody comprises a CDR that differs by 1, 2, 3, or 4 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs: 3, 4, 7, 9, 11, and 13 and does not affect binding affinity by more than 10%, 20%, or 30%.

[0054] In certain embodiments, described herein is an antibody that specifically binds to LIF, comprising a humanized heavy chain comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 49-52; and a humanized light chain comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 53-56. In certain embodiments, described herein is an antibody that specifically binds to LIF, comprising a humanized heavy chain comprising an amino acid sequence set forth in any one of SEQ ID NOs: 49-52; and a humanized light chain comprising the amino acid sequence set forth in any one of SEQ ID NOs: 53-56.

[0055] In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 50; and a humanized light chain comprising an amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 54. In certain embodiments, described herein are antibodies that specifically bind to LIF, comprising a humanized heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 50; and a humanized light chain comprising the amino acid sequence set forth in any one of SEQ ID NOs: 54.

[0056] Epitopes bound by therapeutically useful LIF antibodies Described herein are unique epitopes of human LIF that, when bound, inhibit LIF biological activity (e.g., STAT3 phosphorylation) and inhibit tumor growth in vivo. The epitopes described herein consist of two discontinuous stretches of amino acids (from residues 13 to 32 and from residues 120 to 138 of human LIF) present in two distinct topological domains of the human LIF protein (α-helices A and C). This binding is a combination of weak (van der Waals attraction), moderate (hydrogen bond), and strong (salt bridge) interactions. In certain embodiments, contact residues are residues on LIF that form hydrogen bonds with residues on an anti-LIF antibody. In certain embodiments, contact residues are residues on LIF that form salt bridges with residues on an anti-LIF antibody. In certain embodiments, contact residues are residues on LIF that form van der Waals attraction with residues on an anti-LIF antibody and are within at least 5, 4, or 3 angstroms of residues on the anti-LIF antibody.

[0057] In certain embodiments, the methods and compositions comprising a LIF-binding antibody and cisplatin described herein include an isolated antibody that binds to any one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty of the following residues of SEQ ID NO: 68: A13, I14, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138. In certain embodiments, described herein is an isolated antibody that binds to all of the following residues A13, 114, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138 of SEQ ID NO: 68. In certain embodiments, described herein is an isolated antibody that binds to all of the following residues A13, 114, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138 of SEQ ID NO: 68. In certain embodiments, the antibody binds only to residues that are involved in strong or moderate interactions with the antibody. In certain embodiments, the antibody binds only to residues that are involved in strong interactions with the antibody. In certain embodiments, the antibody interacts with helices A and C of LIF. In certain embodiments, the antibody blocks LIF interaction with gp130.

[0058] In certain embodiments, described herein are antibodies comprising a CDR having an amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 6, 9, 11, and 13 that binds to any one, two, three, four, five, six, seven, eight, nine, ten, eleven, 12, 13, 14, 15, 16, 17, 18, 19, or 20 of the following residues of SEQ ID NO: 68: A13, I14, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138. In certain embodiments, described herein are antibodies comprising a CDR having the amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 6, 9, 11, and 13 that binds to all of the following residues: A13, 114, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138 of SEQ ID NO: 68. In certain embodiments, the antibody binds only to residues that participate in strong or moderate interactions with the antibody. In certain embodiments, the antibody binds only to residues that participate in strong interactions with the antibody.

[0059] In certain embodiments, described herein are antibodies comprising a CDR having an amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 8, 9, 11, and 13 that binds to any one, two, three, four, five, six, seven, eight, nine, ten, eleven, 12, 13, 14, 15, 16, 17, 18, 19, or 20 of the following residues of SEQ ID NO: 68: A13, I14, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138. In certain embodiments, described herein are antibodies comprising a CDR having the amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 8, 9, 11, and 13 that binds to all of the following residues: A13, 114, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138 of SEQ ID NO: 68. In certain embodiments, the antibody binds only to residues that participate in strong or moderate interactions with the antibody. In certain embodiments, the antibody binds only to residues that participate in strong interactions with the antibody.

[0060] In certain embodiments, described herein are antibodies that differ by 1, 2, 3, 4, or 5 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 6, 9, 11, and 13, and that contain CDRs that bind any one, two, three, four, five, six, seven, eight, nine, ten, eleven, 12, 13, 14, 15, 16, 17, 18, 19, or 20 of the following residues of SEQ ID NO: 68: A13, I14, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138. In certain embodiments, described herein are antibodies that differ by 1, 2, 3, 4, or 5 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 6, 9, 11, and 13, and contain CDRs that bind all of the following residues: A13, 114, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138 of SEQ ID NO: 68. In certain embodiments, the antibody binds only to residues that participate in strong or moderate interactions with the antibody. In certain embodiments, the antibody binds only to residues that participate in strong interactions with the antibody.

[0061] In certain embodiments, described herein are antibodies that differ by 1, 2, 3, 4, or 5 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 8, 9, 11, and 13, and that contain CDRs that bind any one, two, three, four, five, six, seven, eight, nine, ten, eleven, 12, 13, 14, 15, 16, 17, 18, 19, or 20 of the following residues of SEQ ID NO: 68: A13, I14, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138. In certain embodiments, described herein are antibodies that differ by 1, 2, 3, 4, or 5 amino acids from the amino acid sequence set forth in any one of SEQ ID NOs: 1, 4, 8, 9, 11, and 13, and contain CDRs that bind all of the following residues: A13, 114, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138 of SEQ ID NO: 68. In certain embodiments, the antibody binds only to residues that participate in strong or moderate interactions with the antibody. In certain embodiments, the antibody binds only to residues that participate in strong interactions with the antibody.

[0062] In certain embodiments, described herein is a humanized antibody that specifically binds to LIF comprising a humanized heavy chain variable region amino acid sequence that is at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO:42; and a humanized light chain variable region amino acid sequence that is at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO:46, and having the following sequence: An antibody that binds to any one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty of residues in sequence number 68: A13, I14, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138. In certain embodiments, described herein are antibodies that specifically bind to LIF comprising a humanized heavy chain variable region amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 42; and a humanized light chain variable region amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 46, and that bind to all of the following residues: A13, I14, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138 of SEQ ID NO: 68. In certain embodiments, the antibody binds only to residues that are involved in strong or moderate interactions with the antibody. In certain embodiments, the antibody binds only to residues that are involved in strong interactions with the antibody.

[0063] In certain embodiments, described herein is a humanized antibody that specifically binds to LIF comprising a humanized heavy chain variable region amino acid sequence that is at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO:66; and a humanized light chain variable region amino acid sequence that is at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO:46, and having the following sequence: An antibody that binds to any one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty of residues in sequence number 68: A13, I14, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138. In certain embodiments, described herein are antibodies that specifically bind to LIF comprising a humanized heavy chain variable region amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 66; and a humanized light chain variable region amino acid sequence at least about 80%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence set forth in SEQ ID NO: 46, and that bind to all of the following residues: A13, I14, R15, H16, P17, C18, H19, N20, Q25, Q29, Q32, D120, R123, S127, N128, L130, C131, C134, S135, or H138 of SEQ ID NO: 68. In certain embodiments, the antibody binds only to residues that are involved in strong or moderate interactions with the antibody. In certain embodiments, the antibody binds only to residues that are involved in strong interactions with the antibody.

[0064] In certain embodiments, the antibodies disclosed herein inhibit LIF signaling in cells. In certain embodiments, the IC for biological inhibition of antibodies under serum starvation conditions in U-251 cells is 50 is about 100, 75, 50, 40, 30, 20, 10, 5, or 1 nanomolar or less. In certain embodiments, the IC for biological inhibition of the antibody under serum-starved conditions in U-251 cells50 is less than or equal to about 900, 800, 700, 600, 500, 400, 300, 200, or 100 nanomolar.

[0065] In certain embodiments, the antibodies disclosed herein are useful for treating tumors and cancers that express LIF. In certain embodiments, individuals treated with the antibodies of this disclosure have been selected for treatment if they have a LIF-positive tumor / cancer. In certain embodiments, the tumor is LIF-positive or produces elevated levels of LIF. In certain embodiments, LIF positivity is determined relative to a baseline or set of pathological criteria. In certain embodiments, LIF-positive tumors express 2-fold, 3-fold, 5-fold, 10-fold, 100-fold, or more LIF than the non-transformed cells from which the tumor originated. In certain embodiments, the tumor has ectopic expression of LIF. LIF-positive tumors can be determined histologically, for example, using immunohistochemistry with anti-LIF antibodies; by commonly used molecular biology methods, for example, mRNA quantification by real-time PCR or RNA-seq; or by protein quantification, for example, by Western blot, flow cytometry, ELISA, or homogeneous protein quantification assays (e.g., AlphaLISA®). In certain embodiments, the antibodies can be used to treat patients diagnosed with cancer, hi certain embodiments, the cancer comprises or is one or more cancer stem cells.

[0066] In certain embodiments, the antibodies disclosed herein are useful for treating tumors in cancers that express the LIF receptor (CD118). LIF receptor-positive tumors can be determined by histopathology or flow cytometry and, in certain embodiments, contain cells that bind to the LIF receptor antibody at 2x, 3x, 3x, 4x, 5x, 10x, or more than the isotype control. In certain embodiments, the tumor has ectopic expression of the LIF receptor. In certain embodiments, the cancer is a cancer stem cell. In certain embodiments, LIF-positive tumors or cancers can be determined by immunohistochemistry using anti-LIF and anti-LIF antibodies. In certain embodiments, LIF-positive tumors are determined by IHC analysis using LIF levels in up to 10%, 20%, 30%, 40%, or 50% of the tumor.

[0067] The antibodies described herein affect a number of outcomes. In certain embodiments, the antibodies described herein can reduce the presence of M2 macrophages in tumors by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to a control antibody (e.g., an isotype control) in tumor models. M2 macrophages can be identified by staining for CCL22 and CD206 in IHC sections or by flow cytometry of tumor-infiltrating immune or myeloid cells. In certain embodiments, the antibodies described herein can reduce LIF binding to gp130 tumors by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to a control antibody (e.g., an isotype control). In certain embodiments, the antibodies described herein can reduce LIF signaling in LIF-responsive cell lines by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to a control antibody (e.g., an isotype control). LIF signaling can be measured, for example, by Western blot against phosphorylated STAT3 (a downstream target of LIF signaling). The antibodies herein are also highly specific for LIF compared to other IL-6 family member cytokines. In certain embodiments, the antibodies bind to human LIF with an affinity that is about 10x, about 50x, or about 100x greater than the affinity of any other IL-6 family member cytokine. In certain embodiments, the LIF antibodies do not bind to other IL-6 family member cytokines produced in mammalian systems. In certain embodiments, the antibodies do not bind to oncostatin M produced in mammalian systems.

[0068] In certain embodiments, LIF-binding polypeptides and antibodies can be administered by any route suitable for administering antibody-containing pharmaceutical compositions, such as subcutaneously, intraperitoneally, intravenously, intramuscularly, intratumorally, or intracerebrally. In certain embodiments, antibodies are administered intravenously. In certain embodiments, antibodies are administered at a suitable dosing schedule (e.g., weekly, twice-weekly, monthly, twice-monthly, etc.). In certain embodiments, antibodies are administered once every three weeks. Antibodies can be administered in any therapeutically effective amount. In certain embodiments, a therapeutically acceptable amount is from about 0.1 mg / kg to about 50 mg / kg. In certain embodiments, a therapeutically acceptable amount is from about 1 mg / kg to about 40 mg / kg. In certain embodiments, a therapeutically acceptable amount is from about 5 mg / kg to about 30 mg / kg. LIF-binding polypeptides or antibodies can be administered intravenously over a period of at least about 60 minutes; however, this period may vary somewhat depending on the conditions of each individual administration.

[0069] Platinum anticancer agents Platinum-based antineoplastic agents, also known as platins, are coordination complexes of platinum used to treat cancer. The LIF-binding polypeptides and antibodies described herein can be combined with platinum-based antineoplastic agents and employed in methods for treating tumors, cancers, or other neoplasms. In certain embodiments, the LIF-binding polypeptides and antibodies described herein can be combined with platinum-based antineoplastic agents in pharmaceutical compositions useful for treating cancers, tumors, or other neoplasms. The h5D8 antibody described herein can be combined with platinum-based antineoplastic agents and employed in methods for treating tumors, cancers, or other neoplasms. In certain embodiments, the h5D8 antibody described herein can be combined with platinum-based antineoplastic agents in pharmaceutical compositions useful for treating cancers, tumors, or other neoplasms.

[0070] The platinum-based anti-cancer agents utilized in the compositions and methods herein can cross-link or bind to DNA. Cross-linking can inhibit DNA repair or DNA synthesis in cancer cells. Cross-linking can disrupt cellular processes and lead to apoptosis. In some embodiments, the platinum-based anti-cancer agent is cisplatin (Platinol®, cis-diamminedichloridoplatinum(II) (CDDP), cis-diammineplatinum(II) dichloride, [SP-4-2]-diamminedichloroplatinum); carboplatin (Paraplatin®, cis-diammine(1,1-cyclobutanedicarboxylato)platinum); oxaliplatin (Eloxatin®, [SP-4-2]-(1R-trans)]-(1,2-cyclohexanediamine-N,N′)[ethanedioata(2-)-O,O′]platinum); nedaplatin (Aqupla®, cis-diammine(glycolato-O,O′)platinum, [SP-4-3]-diammine(hydroxyacetato(2-)-O) 1 ,O 2 ) platinum); triplatin tetranitrate (BBR3464, hexaamminedichloro-bis[μ-(1,6-hexanediamine-κN:κN')]tri-platinum (4 + )); phenatriplatin nitrate ([SP-4-3]-diamminechloro(phenanthridine)-platinum(1+), picoplatin ([SP-4-3]-amminedichloro(2-methylpyridine)platinum), satraplatin (JM216, [OC-6-43]-bis(acetato-O)amminedichloro(cyclohexanamine)platinum); lobaplatin ([SP-4-3-(S),(trans)]-(1,2-cyclobutanedimethanamine-N,N')[2-hydroxypropanoato(2-)-O 1 ,O 2]-platinum; heptaplatin (Eptaplatin, [SP-4-2-[4R-(2α,4α,5β)]]-[2-(1-methylethyl)-1,3-dioxolane-4,5-dimethanamine-N,N'][propanedioato(2-)-O,O']-platinum,); a pharmaceutically acceptable salt thereof, or a combination thereof. In some embodiments, the platinum-based anti-cancer agent is cisplatin.

[0071] In certain embodiments, the platinum-based antineoplastic agent can be administered by any route suitable for administering a small molecule-containing pharmaceutical composition, such as subcutaneously, intraperitoneally, intravenously, intramuscularly, intratumorally, intracerebrally, or orally. In certain embodiments, the platinum-based antineoplastic agent is administered intravenously. In certain embodiments, the platinum-based antineoplastic agent is administered at a suitable dosing schedule, for example, daily, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, weekly, twice weekly, monthly, twice monthly, every 2 weeks, every 3 weeks, or every 4 weeks. The platinum-based antineoplastic agent can be administered in any therapeutically effective amount. In certain embodiments, a therapeutically acceptable amount is about 1 mg / m 2 ~about 1000mg / m 2 (body surface area). In certain embodiments, a therapeutically acceptable amount is about 50 mg / m 2 ~about 70mg / m 2 In certain embodiments, a therapeutically acceptable amount is about 50 mg / m 2 ~about 100mg / m 2 , about 50mg / m 2 ~about 150mg / m 2 , or about 50 mg / m 2 ~about 200mg / m 2 (body surface area). In certain embodiments, a therapeutically acceptable amount is about 10 mg / m 2 ~about 50mg / m 2 , about 10mg / m 2 ~about 70mg / m 2 , or 10 mg / m 2 ~about 100mg / m 2(body surface area). In certain embodiments, a therapeutically acceptable amount is about 1 mg / m 2 ~about 50mg / m 2 , about 1mg / m 2 ~about 70mg / m 2 , or about 1 mg / m 2 ~about 100mg / m 2 (body surface area). In certain embodiments, a therapeutically acceptable amount is about 100 mg / m 2 ~about 200mg / m 2 , about 200mg / m 2 ~about 500mg / m 2 , or about 300 mg / m 2 ~about 600mg / m 2 (body surface area). In certain embodiments, a therapeutically acceptable amount is about 1 mg / m 2 , 2 mg / m 2 , 3 mg / m 2 , 4 mg / m 2 , 5 mg / m 2 , 6 mg / m 2 , 7 mg / m 2 , 8 mg / m 2 , 9 mg / m 2 , 10 mg / m 2 , 20 mg / m 2 , 30 mg / m 2 , 40 mg / m 2 , 50 mg / m 2 , 60 mg / m 2 , 70 mg / m 2 , 80 mg / m 2 , 90 mg / m 2 , 100 mg / m 2 , 110 mg / m 2 , 120 mg / m 2 , 130 mg / m 2 , 140 mg / m 2 , 150 mg / m 2 , 160 mg / m 2 , 170 mg / m 2 , 180 mg / m 2 , 190 mg / m 2 , 200 mg / m 2 , 300 mg / m 2 , 400 mg / m 2 , 500 mg / m 2, 600 mg / m 2 , 700 mg / m 2 , 800 mg / m 2 , 900 mg / m 2 , or 1000 mg / m 2 (body surface area).

[0072] In certain embodiments, cisplatin can be administered in any therapeutically effective amount. In certain embodiments, a therapeutically acceptable amount of cisplatin is about 50 mg / m 2 ~about 70mg / m 2 (body surface area). In certain embodiments, the therapeutically acceptable amount of cisplatin is about 50 mg / m 2 ~about 100mg / m 2 (body surface area). In certain embodiments, the therapeutically acceptable amount of cisplatin is about 10 mg / m 2 ~about 70mg / m 2 (body surface area). In certain embodiments, the therapeutically acceptable amount of cisplatin is about 10 mg / m 2 ~about 100mg / m 2 (body surface area). In certain embodiments, the therapeutically acceptable amount of cisplatin is about 1 mg / m 2 ~about 50mg / m 2 (body surface area). In certain embodiments, the therapeutically acceptable amount of cisplatin is about 1 mg / m 2 ~about 70mg / m 2 (body surface area). In certain embodiments, the therapeutically acceptable amount of cisplatin is about 1 mg / m 2 ~about 100mg / m 2 (body surface area). In certain embodiments, the therapeutically acceptable amount of cisplatin is about 1 mg / m 2 , 2 mg / m 2 , 3 mg / m 2 , 4 mg / m 2 , 5 mg / m 2 , 6 mg / m 2 , 7 mg / m 2 , 8 mg / m 2 , 9 mg / m 2 , 10 mg / m 2 , 20 mg / m2 , 30 mg / m 2 , 40 mg / m 2 , 50 mg / m 2 , 60 mg / m 2 , 70 mg / m 2 , 80 mg / m 2 , 90 mg / m 2 , or 100 mg / m 2 (body surface area).

[0073] In certain embodiments, a platinum-based antineoplastic agent may be administered to an individual at a fixed dosage level of about 10 milligrams to about 1500 milligrams. In certain embodiments, a platinum-based antineoplastic agent may be administered to an individual at a fixed dosage level of about 50 milligrams to about 1000 milligrams, about 50 milligrams to about 800 milligrams, about 50 milligrams to about 600 milligrams, about 50 milligrams to about 500 milligrams, or about 100 milligrams to about 500 milligrams. In certain embodiments, the platinum-based antineoplastic agent administered to an individual may be at a fixed dosage level of about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1100, 1200, 1300, 1400, or 1500 milligrams. In certain embodiments, the platinum-based antineoplastic agent administered to an individual may be at a level suitable for monotherapy.

[0074] In certain embodiments, cisplatin may be administered to an individual at a fixed dosage level of about 10 milligrams to about 500 milligrams. In certain embodiments, cisplatin may be administered to an individual at a fixed dosage level of about 50 milligrams to about 500 milligrams, about 50 milligrams to about 400 milligrams, about 100 milligrams to about 400 milligrams, about 100 milligrams to about 300 milligrams, or about 100 milligrams to about 200 milligrams. In certain embodiments, cisplatin may be administered to an individual at a fixed dosage level of about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, or 500 milligrams. In certain embodiments, cisplatin may be administered to an individual at a level suitable for monotherapy.

[0075] H5D8 Dosage In certain embodiments, the antibody can be administered by any route suitable for administering an antibody-containing pharmaceutical composition, such as, for example, subcutaneously, intraperitoneally, intravenously, intramuscularly, intratumorally, or intracerebrally. In certain embodiments, the antibody is administered intravenously. In certain embodiments, the antibody is administered on a suitable dosing schedule (e.g., weekly, twice weekly, monthly, twice monthly, etc.). In certain embodiments, the antibody is administered once every three weeks. The antibody can be administered in any therapeutically effective amount. In certain embodiments, a therapeutically acceptable amount is about 0.01 mg / kg to about 50 mg / kg. In certain embodiments, a therapeutically acceptable amount is about 0.05 mg / kg to about 50 mg / kg. In certain embodiments, a therapeutically acceptable amount is about 0.1 mg / kg to about 50 mg / kg. In certain embodiments, a therapeutically acceptable amount is about 1 mg / kg to about 40 mg / kg. In certain embodiments, a therapeutically acceptable amount is about 5 mg / kg to about 30 mg / kg. The h5D8 antibody can be administered at a fixed dose regardless of the weight or mass of the individual to whom the h5D8 antibody is administered. The h5D8 antibody can be administered at a fixed dose regardless of the weight or mass of the individual to whom the h5D8 antibody is administered, provided that the individual has a mass of at least about 37.5 kilograms. A fixed dose of h5D8 from about 1 milligram to about 2000 milligrams can be administered. A fixed dose of h5D8 from about 10 milligrams to about 2000 milligrams can be administered. A fixed dose of h5D8 from about 25 milligrams to about 2000 milligrams can be administered. A fixed dose of h5D8 from about 50 milligrams to about 2000 milligrams can be administered. A fixed dose of h5D8 from about 75 milligrams to about 2000 milligrams can be administered. A fixed dose of h5D8 can be administered from about 225 milligrams to about 2000 milligrams, from about 750 milligrams to about 2000 milligrams, from about 1125 milligrams to about 2000 milligrams, or from about 1500 milligrams to about 2000 milligrams. A fixed dose of h5D8 of about 1 milligram can be administered. A fixed dose of h5D8 of about 10 milligrams can be administered.A fixed dose of about 25 milligrams of h5D8 can be administered. A fixed dose of about 50 milligrams of h5D8 can be administered. A fixed dose of about 75 milligrams of h5D8 can be administered. A fixed dose of about 225 milligrams of h5D8 can be administered. A fixed dose of about 750 milligrams of h5D8 can be administered. A fixed dose of about 1125 milligrams of h5D8 can be administered. A fixed dose of about 1500 milligrams of h5D8 can be administered. A fixed dose of about 2000 milligrams of h5D8 can be administered.

[0076] Other dosages of h5D8 are also contemplated. A fixed dose of 1275, 1300, 1325, 1350, 1375, 1400, 1425, 1450, 1475, 1525, 1550, 1575, 1600, 1625, 1650, 1675, 1700, 1725, 1750, 1775, 1800, 1825, 1850, 1875, 1900, 1925, 1950, 1975, 2025, 2050, 2075, or 2100 milligrams of h5D8 can be administered. Any of these doses can be administered weekly, every two weeks, every three weeks, or every four weeks.

[0077] A fixed dose of h5D8 from about 1 milligram to about 2000 milligrams can be administered weekly. A fixed dose of h5D8 from about 10 milligrams to about 2000 milligrams can be administered weekly. A fixed dose of h5D8 from about 25 milligrams to about 2000 milligrams can be administered weekly. A fixed dose of h5D8 from about 50 milligrams to about 2000 milligrams can be administered weekly. A fixed dose of h5D8 from about 75 milligrams to about 2000 milligrams can be administered weekly. A fixed dose of h5D8 from about 75 milligrams to about 1500 milligrams can be administered weekly. A fixed dose of about 225 milligrams to about 1500 milligrams, about 750 milligrams to about 1500 milligrams, or about 1125 milligrams to about 1500 milligrams of h5D8 can be administered weekly. A fixed dose of about 1 milligram of h5D8 can be administered weekly. A fixed dose of about 10 milligrams of h5D8 can be administered weekly. A fixed dose of about 25 milligrams of h5D8 can be administered weekly. A fixed dose of about 50 milligrams of h5D8 can be administered weekly. A fixed dose of about 75 milligrams of h5D8 can be administered weekly. A fixed dose of about 225 milligrams of h5D8 can be administered weekly. A fixed dose of about 750 milligrams of h5D8 can be administered weekly. A fixed dose of about 1125 milligrams of h5D8 can be administered once a week. A fixed dose of about 1500 milligrams of h5D8 can be administered once a week. A fixed dose of about 2000 milligrams of h5D8 can be administered once a week.

[0078] A fixed dose of h5D8 from about 1 milligram to about 2000 milligrams can be administered every two weeks. A fixed dose of h5D8 from about 10 milligrams to about 2000 milligrams can be administered every two weeks. A fixed dose of h5D8 from about 25 milligrams to about 2000 milligrams can be administered every two weeks. A fixed dose of h5D8 from about 50 milligrams to about 2000 milligrams can be administered every two weeks. A fixed dose of h5D8 from about 75 milligrams to about 2000 milligrams can be administered every two weeks. A fixed dose of h5D8 from about 75 milligrams to about 1500 milligrams can be administered every two weeks. A fixed dose of about 225 milligrams to about 1500 milligrams, about 750 milligrams to about 1500 milligrams, or about 1125 milligrams to about 1500 milligrams of h5D8 can be administered every two weeks. A fixed dose of about 1 milligram of h5D8 can be administered every two weeks. A fixed dose of about 10 milligrams of h5D8 can be administered every two weeks. A fixed dose of about 25 milligrams of h5D8 can be administered every two weeks. A fixed dose of about 50 milligrams of h5D8 can be administered every two weeks. A fixed dose of about 75 milligrams of h5D8 can be administered every two weeks. A fixed dose of about 225 milligrams of h5D8 can be administered every two weeks. A fixed dose of about 750 milligrams of h5D8 can be administered every two weeks. A fixed dose of about 1125 milligrams of h5D8 can be administered every two weeks. A fixed dose of about 1500 milligrams of h5D8 can be administered every two weeks. A fixed dose of about 2000 milligrams of h5D8 can be administered every two weeks.

[0079] A fixed dose of h5D8 from about 1 milligram to about 2000 milligrams can be administered every three weeks. A fixed dose of h5D8 from about 10 milligrams to about 2000 milligrams can be administered every three weeks. A fixed dose of h5D8 from about 25 milligrams to about 2000 milligrams can be administered every three weeks. A fixed dose of h5D8 from about 50 milligrams to about 2000 milligrams can be administered every three weeks. A fixed dose of h5D8 from about 75 milligrams to about 2000 milligrams can be administered every three weeks. A fixed dose of h5D8 from about 75 milligrams to about 1500 milligrams can be administered every three weeks. A fixed dose of about 225 milligrams to about 1500 milligrams, about 750 milligrams to about 1500 milligrams, or about 1125 milligrams to about 1500 milligrams of h5D8 can be administered every three weeks. A fixed dose of about 1 milligram of h5D8 can be administered every three weeks. A fixed dose of about 10 milligrams of h5D8 can be administered every three weeks. A fixed dose of about 25 milligrams of h5D8 can be administered every three weeks. A fixed dose of about 50 milligrams of h5D8 can be administered every three weeks. A fixed dose of about 75 milligrams of h5D8 can be administered every three weeks. A fixed dose of about 225 milligrams of h5D8 can be administered every three weeks. A fixed dose of about 750 milligrams of h5D8 can be administered every three weeks. A fixed dose of about 1125 milligrams of h5D8 can be administered every three weeks. A fixed dose of about 1500 milligrams of h5D8 can be administered every three weeks. A fixed dose of about 2000 milligrams of h5D8 can be administered every three weeks.

[0080] A fixed dose of h5D8 from about 1 milligram to about 2000 milligrams can be administered every four weeks. A fixed dose of h5D8 from about 10 milligrams to about 2000 milligrams can be administered every four weeks. A fixed dose of h5D8 from about 25 milligrams to about 2000 milligrams can be administered every four weeks. A fixed dose of h5D8 from about 50 milligrams to about 2000 milligrams can be administered every four weeks. A fixed dose of h5D8 from about 75 milligrams to about 2000 milligrams can be administered every four weeks. A fixed dose of h5D8 from about 75 milligrams to about 1500 milligrams can be administered every four weeks. A fixed dose of about 225 milligrams to about 1500 milligrams, about 750 milligrams to about 1500 milligrams, or about 1125 milligrams to about 1500 milligrams of h5D8 can be administered every four weeks. A fixed dose of about 1 milligram of h5D8 can be administered every four weeks. A fixed dose of about 10 milligrams of h5D8 can be administered every four weeks. A fixed dose of about 25 milligrams of h5D8 can be administered every four weeks. A fixed dose of about 50 milligrams of h5D8 can be administered every four weeks. A fixed dose of about 75 milligrams of h5D8 can be administered every four weeks. A fixed dose of about 225 milligrams of h5D8 can be administered every four weeks. A fixed dose of about 750 milligrams of h5D8 can be administered every four weeks. A fixed dose of about 1125 milligrams of h5D8 can be administered once every four weeks. A fixed dose of about 1500 milligrams of h5D8 can be administered once every four weeks. A fixed dose of about 2000 milligrams of h5D8 can be administered once every four weeks.

[0081] The h5D8 antibody can be administered at a dose based on the weight or mass of the individual to whom the h5D8 antibody is administered. A weight-adjusted dose of about 0.01 mg / kg to about 25 mg / kg of h5D8 can be administered. A weight-adjusted dose of about 0.05 mg / kg to about 25 mg / kg of h5D8 can be administered. A weight-adjusted dose of about 0.1 mg / kg to about 25 mg / kg of h5D8 can be administered. A weight-adjusted dose of about 0.5 mg / kg to about 25 mg / kg of h5D8 can be administered. A weight-adjusted dose of about 1 mg / kg to about 25 mg / kg of h5D8 can be administered. A body weight-adjusted dose of h5D8 of about 3 mg / kg to about 25 mg / kg, about 10 mg / kg to about 25 mg / kg, about 15 mg / kg to about 25 mg / kg, or about 20 mg / kg to about 25 mg / kg can be administered. A body weight-adjusted dose of h5D8 of about 1 mg / kg can be administered. A body weight-adjusted dose of h5D8 of about 3 mg / kg can be administered. A body weight-adjusted dose of h5D8 of about 10 mg / kg can be administered. A body weight-adjusted dose of h5D8 of about 15 mg / kg can be administered. A body weight-adjusted dose of h5D8 of about 20 mg / kg can be administered. A body weight-adjusted dose of h5D8 of about 25 mg / kg can be administered.

[0082] A weight-adjusted dose of h5D8 from about 0.01 mg / kg to about 25 mg / kg can be administered once every three weeks. A weight-adjusted dose of h5D8 from about 0.05 mg / kg to about 25 mg / kg can be administered once every three weeks. A weight-adjusted dose of h5D8 from about 0.1 mg / kg to about 25 mg / kg can be administered once every three weeks. A weight-adjusted dose of h5D8 from about 0.5 mg / kg to about 25 mg / kg can be administered once every three weeks. A weight-adjusted dose of h5D8 from about 1 mg / kg to about 25 mg / kg can be administered once every three weeks. A weight-adjusted dose of h5D8 from about 3 mg / kg to about 25 mg / kg, from about 10 mg / kg to about 20 mg / kg, from about 15 mg / kg to about 25 mg / kg, or from about 20 mg / kg to about 25 mg / kg can be administered once a week, every two weeks, every three weeks, or every four weeks.

[0083] Other weight-adjusted doses of h5D8 are also contemplated. A weight-adjusted dose of about 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 21 mg / kg, 22 mg / kg, 23 mg / kg, 24 mg / kg, 26 mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, or 30 mg / kg of h5D8 can be administered. Any of these doses can be administered weekly, every two weeks, every three weeks, or every four weeks.

[0084] A weight-adjusted dose of about 0.01 mg / kg of h5D8 can be administered weekly. A weight-adjusted dose of about 0.05 mg / kg of h5D8 can be administered weekly. A weight-adjusted dose of about 0.1 mg / kg of h5D8 can be administered weekly. A weight-adjusted dose of about 0.5 mg / kg of h5D8 can be administered weekly. A weight-adjusted dose of about 1 mg / kg of h5D8 can be administered weekly. A weight-adjusted dose of about 3 mg / kg of h5D8 can be administered weekly. A weight-adjusted dose of about 10 mg / kg of h5D8 can be administered weekly. A weight-adjusted dose of about 15 mg / kg of h5D8 can be administered weekly. A weight-adjusted dose of about 20 mg / kg of h5D8 can be administered once a week. A weight-adjusted dose of about 25 mg / kg of h5D8 can be administered once a week.

[0085] A weight-adjusted dose of about 0.01 mg / kg of h5D8 can be administered every two weeks. A weight-adjusted dose of about 0.05 mg / kg of h5D8 can be administered every two weeks. A weight-adjusted dose of about 0.1 mg / kg of h5D8 can be administered every two weeks. A weight-adjusted dose of about 0.5 mg / kg of h5D8 can be administered every two weeks. A weight-adjusted dose of about 1 mg / kg of h5D8 can be administered every two weeks. A weight-adjusted dose of about 3 mg / kg of h5D8 can be administered every two weeks. A weight-adjusted dose of about 10 mg / kg of h5D8 can be administered every two weeks. A weight-adjusted dose of about 15 mg / kg of h5D8 can be administered every two weeks. A weight-adjusted dose of about 20 mg / kg of h5D8 can be administered once every two weeks. A weight-adjusted dose of about 25 mg / kg of h5D8 can be administered once every two weeks.

[0086] A weight-adjusted dose of about 0.01 mg / kg of h5D8 can be administered every three weeks. A weight-adjusted dose of about 0.05 mg / kg of h5D8 can be administered every three weeks. A weight-adjusted dose of about 0.1 mg / kg of h5D8 can be administered every three weeks. A weight-adjusted dose of about 0.5 mg / kg of h5D8 can be administered every three weeks. A weight-adjusted dose of about 1 mg / kg of h5D8 can be administered every three weeks. A weight-adjusted dose of about 3 mg / kg of h5D8 can be administered every three weeks. A weight-adjusted dose of about 10 mg / kg of h5D8 can be administered every three weeks. A weight-adjusted dose of about 15 mg / kg of h5D8 can be administered every three weeks. A weight-adjusted dose of about 20 mg / kg of h5D8 can be administered once every three weeks. A weight-adjusted dose of about 25 mg / kg of h5D8 can be administered once every three weeks.

[0087] A weight-adjusted dose of about 0.01 mg / kg of h5D8 can be administered once every four weeks. A weight-adjusted dose of about 0.05 mg / kg of h5D8 can be administered once every four weeks. A weight-adjusted dose of about 0.1 mg / kg of h5D8 can be administered once every four weeks. A weight-adjusted dose of about 0.5 mg / kg of h5D8 can be administered once every four weeks. A weight-adjusted dose of about 1 mg / kg of h5D8 can be administered once every four weeks. A weight-adjusted dose of about 3 mg / kg of h5D8 can be administered once every four weeks. A weight-adjusted dose of about 10 mg / kg of h5D8 can be administered once every four weeks. A weight-adjusted dose of about 15 mg / kg of h5D8 can be administered once every four weeks. A weight-adjusted dose of about 20 mg / kg of h5D8 can be administered once every four weeks. A weight-adjusted dose of about 25 mg / kg of h5D8 can be administered once every four weeks.

[0088] Any of the doses detailed herein can be administered iv over a period of at least about 60 minutes; however, this period may vary somewhat based on the conditions for each individual administration.

[0089] Combination therapy dosing schedule Combination therapy comprising a LIF-binding polypeptide and a platinum-based antineoplastic agent can be administered in various ways. The LIF-binding polypeptide and the platinum-based antineoplastic agent can be administered simultaneously on the same schedule, or at different times on different schedules. When administered simultaneously, administration can be via separate formulations or a single formulation containing both the LIF-binding polypeptide and the platinum-based antineoplastic agent. The modes of administration can be mixed; for example, the LIF-binding polypeptide can be administered intravenously, and the platinum-based antineoplastic agent can be administered simultaneously or orally or by parenteral injection. In certain embodiments, the LIF-binding polypeptide is administered intravenously, parenterally, subcutaneously, intratumorally, or orally. In certain embodiments, the platinum-based antineoplastic agent is administered intravenously, parenterally, subcutaneously, intratumorally, or orally.

[0090] When the combination therapy is administered to an individual on the same schedule, the LIF-binding polypeptide and the platinum-based antineoplastic agent can be administered once a week, once every two weeks, or once every four weeks. The LIF-binding polypeptide and the platinum-based antineoplastic agent can be administered separately or as a single formulation.

[0091] When the combination therapy is administered to an individual on different schedules, the LIF-binding polypeptide and the platinum-based antineoplastic agent can be administered alternately. In certain embodiments, the platinum-based antineoplastic agent can be administered to an individual one or more times prior to administration of the LIF-binding polypeptide. The LIF-binding polypeptide can be administered within 1, 2, 3, 4, 5, or 6 days after administration of the platinum-based antineoplastic agent. The LIF-binding polypeptide can be administered within 1, 2, 3, or 4 weeks after administration of the platinum-based antineoplastic agent. The h5D8 antibody can be administered within 1, 2, 3, 4, 5, or 6 days after administration of the platinum-based antineoplastic agent. The h5D8 antibody can be administered within 1, 2, 3, or 4 weeks after administration of the platinum-based antineoplastic agent.

[0092] The LIF-binding polypeptide can be administered to an individual one or more times prior to administration of the platinum-based antineoplastic agent. In certain embodiments, the platinum-based antineoplastic agent can be administered within 1 day, 2 days, 3 days, 4 days, 5 days, or 6 days after administration of the LIF-binding polypeptide. In certain embodiments, the platinum-based antineoplastic agent can be administered within 1 week, 2 weeks, 3 weeks, or 4 weeks after administration of the LIF-binding polypeptide. In certain embodiments, the platinum-based antineoplastic agent can be administered within 1 day, 2 days, 3 days, 4 days, 5 days, or 6 days after administration of the h5D8 antibody. In certain embodiments, the platinum-based antineoplastic agent can be administered within 1 week, 2 weeks, 3 weeks, or 4 weeks after administration of the h5D8 antibody.

[0093] In certain embodiments, the LIF-binding polypeptide can be administered to an individual once a week, and the platinum-based antineoplastic agent can be administered to the individual every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the LIF-binding polypeptide can be administered to an individual once every two weeks, and the platinum-based antineoplastic agent can be administered to the individual every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the LIF-binding polypeptide can be administered to an individual once every three weeks, and the platinum-based antineoplastic agent can be administered to the individual every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the LIF-binding polypeptide can be administered to an individual once every four weeks, and the platinum-based antineoplastic agent can be administered to the individual every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the platinum-based antineoplastic agent can be administered to an individual once a week, and the LIF-binding polypeptide can be administered to an individual every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the platinum-based antineoplastic agent can be administered to an individual once every two weeks, and the LIF-binding polypeptide can be administered to an individual every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the platinum-based antineoplastic agent can be administered to an individual once every three weeks, and the LIF-binding polypeptide can be administered to an individual every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the platinum-based antineoplastic agent can be administered to an individual once every four weeks, and the LIF-binding polypeptide can be administered to an individual every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, h5D8 can be administered to an individual one or more times prior to administration of the platinum-based antineoplastic agent. In certain embodiments, h5D8 can be administered to an individual once a week, and a platinum-based antineoplastic agent can be administered to an individual every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, h5D8 can be administered to an individual once every two weeks, and a platinum-based antineoplastic agent can be administered to an individual every week, every two weeks, every three weeks, or every four weeks.In certain embodiments, h5D8 can be administered to an individual once every three weeks, and a platinum-based antineoplastic agent can be administered to an individual every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, h5D8 can be administered to an individual once every four weeks, and a platinum-based antineoplastic agent can be administered to an individual every week, every two weeks, every three weeks, or every four weeks.

[0094] Combination therapies according to the present disclosure can include combinations in which one or both of the active components (e.g., a LIF-binding polypeptide and a platinum-based antineoplastic agent) are ineffective alone as monotherapy but are effective when administered as part of a combination therapy. In certain embodiments, the platinum-based antineoplastic agent is administered at a level that is ineffective as a monotherapy but is effective in combination with a LIF-binding polypeptide. In certain embodiments, the platinum-based antineoplastic agent is administered at a level that is ineffective as a monotherapy but is effective in combination with an h5D8 antibody. In certain embodiments, the LIF-binding polypeptide is administered at a level that is ineffective as a monotherapy but is effective in combination with a platinum-based antineoplastic agent. In certain embodiments, h5D8 is administered at a level that is ineffective as a monotherapy but is effective in combination with a platinum-based antineoplastic agent. In certain embodiments, both the LIF-binding polypeptide and the platinum-based antineoplastic agent are administered at levels that are ineffective as a monotherapy but are effective in combination. In certain embodiments, both h5D8 and the platinum-based anti-neoplastic agent are administered at levels that are not effective as monotherapy, but are effective in combination.

[0095] Treatment indications In certain embodiments, disclosed herein are methods and compositions useful for treating cancer or tumors. In certain embodiments, cancers include tumors of the breast, heart, lung, small intestine, colon, spleen, kidney, bladder, head, neck, ovary, prostate, brain, pancreas, skin, bone, bone marrow, blood, thymus, uterus, testis, and liver. In certain embodiments, tumors that can be treated with the antibodies of the invention include adenoma, adenocarcinoma, angiosarcoma, astrocytoma, epithelial carcinoma, germinoma, glioblastoma, glioma, hemangioendothelioma, angiosarcoma, hematoma, hepatoblastoma, leukemia, lymphoma, medulloblastoma, melanoma, neuroblastoma, osteosarcoma, retinoblastoma, rhabdomyosarcoma, sarcoma, and / or teratoma. In certain embodiments, the tumor / cancer is selected from the group consisting of acral lentiginous melanoma, actinic keratosis, adenocarcinoma, adenoid cystic carcinoma, adenoma, adenosarcoma, adenosquamous carcinoma, astrocytic tumor, Bartholin's adenocarcinoma, basal cell carcinoma, bronchial adenocarcinoma, capillary carcinoid, carcinoma, carcinosarcoma, cholangiocarcinoma, chondrosarcoma, cystadenoma, endodermal sinus tumor, endometrial hyperplasia, endometrial stromal sarcoma, endometrioid adenocarcinoma, ependymal sarcoma, Swing's sarcoma, focal nodular hyperplasia, gastronoma, germ line tumor, glioblastoma, glucagonoma, hemangioblastoma, hemangioendothelioma, hemangioma, hepatic adenoma, hepatocellular carcinoma, insulinite, intraepithelial neoplasia, intraepithelial squamous cell carcinoma Squamous cell neoplasia, invasive squamous cell carcinoma, large cell carcinoma, liposarcoma, lung cancer, lymphoblastic leukemia, lymphocytic leukemia, leiomyosarcoma, melanoma, malignant melanoma, malignant mesothelioma, schwannoma, medulloblastoma, medulloepithelioma, mesothelioma, mucoepidermoid carcinoma, myeloid leukemia, neuroblastoma, neuroepithelial adenocarcinoma, nodular melanoma, osteosarcoma, ovarian cancer, serous papillary adenocarcinoma, pituitary tumor, plasmacytoma, pseudosarcoma, prostate cancer, pulmonary blastoma, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, serous carcinoma, squamous cell carcinoma, small cell carcinoma, soft tissue carcinoma, somatostatin-secreting tumor tumor), squamous cell carcinoma, squamous cell carcinoma, undifferentiated carcinoma, uveal melanoma, verrucous carcinoma, vaginal / vulvar cancer, VIPoma, and Wilms' tumor.In certain embodiments, tumors / cancers to be treated with one or more antibodies of the present disclosure include brain cancer, head and neck cancer, colorectal cancer, acute myeloid leukemia, pre-B-cell acute lymphoblastic leukemia, bladder cancer, astrocytoma, preferably grade II, III, or IV astrocytoma, glioblastoma, glioblastoma multiforme, small cell carcinoma, and non-small cell carcinoma, preferably non-small cell lung cancer, lung adenocarcinoma, metastatic melanoma, androgen-independent metastatic prostate cancer, androgen-dependent metastatic prostate cancer, prostate adenocarcinoma, and breast cancer, preferably ductal carcinoma and / or breast carcinoma. In certain embodiments, a cancer treated with an antibody of this disclosure includes glioblastoma. In certain embodiments, a cancer treated with one or more antibodies of this disclosure includes pancreatic cancer. In certain embodiments, a cancer treated with one or more antibodies of this disclosure includes ovarian cancer. In certain embodiments, a cancer treated with one or more antibodies of this disclosure includes lung cancer. In certain embodiments, the cancer treated with one or more antibodies of this disclosure comprises prostate cancer. In certain embodiments, the cancer treated with one or more antibodies of this disclosure comprises colon cancer. In certain embodiments, the cancer treated comprises glioblastoma, pancreatic cancer, ovarian cancer, colon cancer, prostate cancer, or lung cancer. In certain embodiments, the cancer is resistant to other treatments. In certain embodiments, the cancer treated is recurrent. In certain embodiments, the cancer is relapsed / resistant glioblastoma, pancreatic cancer, ovarian cancer, colon cancer, prostate cancer, or lung cancer. In certain embodiments, the cancer comprises advanced solid tumors, glioblastoma, gastric cancer, skin cancer, prostate cancer, pancreatic cancer, breast cancer, testicular cancer, thyroid cancer, head and neck cancer, liver cancer, kidney cancer, esophageal cancer, ovarian cancer, colon cancer, lung cancer, lymphoma, or soft tissue cancer. In certain embodiments, the cancer comprises non-small cell lung cancer, ovarian epithelial cancer, or pancreatic adenocarcinoma. In certain embodiments, the cancer comprises advanced solid tumors. In certain embodiments, the individual is refractory to previous treatment with a LIF-binding antibody as monotherapy. In certain embodiments, the individual is refractory to previous treatment with a platinum-based anti-neoplastic agent as monotherapy.

[0096] Pharmaceutically Acceptable Excipients, Carriers, and Diluents In certain embodiments, the platinum-based antineoplastic agent of the present disclosure and the LIF-binding polypeptide are components of a pharmaceutical composition. In certain embodiments, the platinum-based antineoplastic agent of the present disclosure and the LIF-binding polypeptide are components of the same pharmaceutical composition. In certain embodiments, the pharmaceutical composition comprises a physiologically appropriate salt concentration (e.g., NaCl). In certain embodiments, the pharmaceutical composition comprises about 0.6% to 1.2% NaCl. In certain embodiments, the pharmaceutical composition comprises about 0.7% to 1.1% NaCl. In certain embodiments, the pharmaceutical composition comprises about 0.8% to 1.0% NaCl. In certain embodiments, the pharmaceutical composition further comprises one or more of the following: a buffer, e.g., acetic acid, citric acid, histidine, succinic acid, phosphate, bicarbonate, and hydroxymethylaminomethane (Tris); a surfactant, e.g., polysorbate 80 (Tween 80), polysorbate 20 (Tween 20), polysorbate, and poloxamer 188; a polyol / disaccharide / polysaccharide, e.g., glucose, dextrose, mannose, mannitol, sorbitol, sucrose, trehalose, and dextran 40; an amino acid, e.g., histidine, glycine, or arginine; an antioxidant, e.g., ascorbic acid, methionine; and a chelating agent, e.g., EGTA or EGTA.

[0097] In certain embodiments, the platinum-based antineoplastic agent, the LIF-binding polypeptide, or both the platinum-based antineoplastic agent and the LIF-binding polypeptide of the present disclosure are administered suspended in a sterile solution. In certain embodiments, the platinum-based antineoplastic agent and the LIF-binding polypeptide are administered from the same solution. In certain embodiments, the solution comprises a physiologically appropriate salt concentration (e.g., NaCl). In certain embodiments, the solution comprises about 0.6% to 1.2% NaCl. In certain embodiments, the solution comprises about 0.7% to 1.1% NaCl. In certain embodiments, the solution comprises about 0.8% to 1.0% NaCl. In certain embodiments, a highly concentrated stock solution of an antibody can be diluted to about 0.9% NaCl. In certain embodiments, the solution comprises about 0.9% NaCl. In certain embodiments, the solution further comprises one or more of the following: a buffer solution, e.g., acetic acid, citric acid, histidine, succinic acid, phosphate, bicarbonate, and hydroxymethylaminomethane (Tris); a surfactant, e.g., polysorbate 80 (Tween 80), polysorbate 20 (Tween 20), polysorbate, and poloxamer 188; a polyol / disaccharide / polysaccharide, e.g., glucose, dextrose, mannose, mannitol, sorbitol, sucrose, trehalose, and dextran 40; an amino acid, e.g., histidine, glycine, or arginine; an antioxidant, e.g., ascorbic acid, methionine; and a chelating agent, e.g., EGTA or EGTA. In certain embodiments, the platinum-based antineoplastic agents and LIF-binding polypeptides of the present disclosure are transported / stored lyophilized and reconstituted prior to administration. In certain embodiments, the lyophilized antibody formulation comprises a bulking agent such as mannitol, sorbitol, sucrose, trehalose, and dextran 40. In certain embodiments, the anti-LIF antibodies of the present disclosure can be shipped and stored as a concentrated stock solution that is diluted at the point of treatment for use. In certain embodiments, this stock solution comprises about 25 mM histidine, about 6% sucrose, about 0.01% polysorbate, and about 20 mg / mL anti-LIF antibody. In certain embodiments, the pH of this solution is about 6.0.In certain embodiments, the form administered to an individual is an aqueous solution containing about 25 mM histidine, about 6% sucrose, about 0.01% polysorbate 80, and about 20 mg / mL h5D8 antibody. In certain embodiments, the pH of this solution is about 6.0.

[0098] In certain embodiments, the platinum-based antineoplastic agent and LIF-binding polypeptide of the present disclosure are administered suspended in a sterile solution. In certain embodiments, the platinum-based antineoplastic agent and LIF-binding polypeptide are administered from the same solution. In certain embodiments, the platinum-based antineoplastic agent and LIF-binding polypeptide are administered from separate solutions. In certain embodiments, the solution comprises a physiologically relevant salt concentration (e.g., NaCl). In certain embodiments, the solution comprises about 0.6% to 1.2% NaCl. In certain embodiments, the solution comprises about 0.7% to 1.1% NaCl. In certain embodiments, the solution comprises about 0.8% to 1.0% NaCl. In certain embodiments, a highly concentrated stock solution of an antibody can be diluted to about 0.9% NaCl. In certain embodiments, the solution comprises about 0.9% NaCl. In certain embodiments, the solution further comprises one or more of the following: a buffer solution, e.g., acetic acid, citric acid, histidine, succinic acid, phosphate, bicarbonate, and hydroxymethylaminomethane (Tris); a surfactant, e.g., polysorbate 80 (Tween 80), polysorbate 20 (Tween 20), polysorbate, and poloxamer 188; a polyol / disaccharide / polysaccharide, e.g., glucose, dextrose, mannose, mannitol, sorbitol, sucrose, trehalose, and dextran 40; an amino acid, e.g., histidine, glycine, or arginine; an antioxidant, e.g., ascorbic acid, methionine; and a chelating agent, e.g., EGTA or EGTA. In certain embodiments, the platinum-based antineoplastic agents and LIF-binding polypeptides of the present disclosure are transported / stored lyophilized and reconstituted prior to administration. In certain embodiments, the lyophilized antibody formulation contains bulking agents such as mannitol, sorbitol, sucrose, trehalose, and dextran 40. In certain embodiments, the anti-LIF antibodies of the present disclosure can be shipped and stored as a concentrated stock solution that is diluted at the point of use. In certain embodiments, this stock solution contains about 25 mM histidine, about 6% sucrose, about 0.01% polysorbate, and about 20 mg / mL of anti-LIF antibody. In certain embodiments, the pH of this solution is about 6.0.In certain embodiments, the form administered to an individual is an aqueous solution containing about 25 mM histidine, about 6% sucrose, about 0.01% polysorbate 80, and about 20 mg / mL h5D8 antibody. In certain embodiments, the pH of this solution is about 6.0.

[0099] Also described herein are kits for carrying out the combination therapies described herein. In certain embodiments, the kits include a LIF-binding polypeptide and a platinum-based anti-cancer agent. In certain embodiments, the kits include h5D8 and a platinum-based anti-cancer agent. One or both of the components may be contained in a vial of glass or other suitable material in lyophilized or liquid form.

[0100] In certain embodiments, described herein is the use of a LIF-binding polypeptide in combination with a platinum-based anti-neoplastic agent to treat non-small lung cancer, ovarian epithelial cancer, or pancreatic adenocarcinoma in an individual. In certain embodiments, the LIF-binding polypeptide and the platinum-based anti-neoplastic agent are administered to the individual in separate formulations. In certain embodiments, the LIF-binding polypeptide and the platinum-based anti-neoplastic agent are administered to the individual in the same formulation. In certain embodiments, the LIF-binding polypeptide is administered to the individual before the platinum-based anti-neoplastic agent is administered to the individual. In certain embodiments, the platinum-based anti-neoplastic agent is administered to the individual before the LIF-binding polypeptide is administered to the individual. In certain embodiments, the platinum-based anti-neoplastic agent is administered to the individual simultaneously with the LIF-binding polypeptide. In certain embodiments, the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. In certain embodiments, the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. In certain embodiments, an antibody that specifically binds to LIF comprises at least one framework region derived from a human antibody framework region. In certain embodiments, an antibody that specifically binds to LIF is humanized. In certain embodiments, an antibody that specifically binds to LIF is deimmunized. In certain embodiments, an antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. In certain embodiments, an antibody that specifically binds to LIF is an IgG antibody. In certain embodiments, an antibody that specifically binds to LIF is a Fab, F(ab)2, single domain antibody, single chain variable fragment (scFv), or nanobody.In certain embodiments, an antibody that specifically binds to LIF comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; (d) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13. In certain embodiments, an antibody that specifically binds LIF comprises (a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and (b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO:42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO:46.In certain embodiments, an antibody that specifically binds to LIF comprises (a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60 or 67; and (b) an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. In certain embodiments, an antibody that specifically binds to LIF has a K of less than about 200 picomolar. D In certain embodiments, antibodies that specifically bind to LIF have a K of less than about 100 picomolar. D In certain embodiments, the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriplatin, picoplatin, satraplatin, or a combination thereof. In certain embodiments, the cancer is resistant to treatment with a therapeutic amount of the LIF-binding polypeptide or platinum-based anti-cancer agent administered as monotherapy.

[0101] In certain embodiments, described herein is the use of a LIF-binding polypeptide in combination with a platinum-based antineoplastic agent to treat colorectal cancer in an individual. In certain embodiments, the LIF-binding polypeptide and the platinum-based antineoplastic agent are administered to the individual in separate formulations. In certain embodiments, the LIF-binding polypeptide and the platinum-based antineoplastic agent are administered to the individual in the same formulation. In certain embodiments, the LIF-binding polypeptide is administered to the individual before the platinum-based antineoplastic agent is administered to the individual. In certain embodiments, the platinum-based antineoplastic agent is administered to the individual before the LIF-binding polypeptide is administered to the individual. In certain embodiments, the platinum-based antineoplastic agent is administered to the individual simultaneously with the LIF-binding polypeptide. In certain embodiments, the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. In certain embodiments, the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. In certain embodiments, the antibody that specifically binds to LIF comprises at least one framework region derived from a human antibody framework region. In certain embodiments, an antibody that specifically binds to LIF is humanized. In certain embodiments, an antibody that specifically binds to LIF is deimmunized. In certain embodiments, an antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. In certain embodiments, an antibody that specifically binds to LIF is an IgG antibody. In certain embodiments, an antibody that specifically binds to LIF is a Fab, F(ab)2, single domain antibody, single chain variable fragment (scFv), or nanobody.In certain embodiments, an antibody that specifically binds to LIF comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; (d) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13. In certain embodiments, an antibody that specifically binds LIF comprises (a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and (b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO:42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO:46.In certain embodiments, an antibody that specifically binds to LIF comprises (a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60 or 67; and (b) an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. In certain embodiments, an antibody that specifically binds to LIF has a K of less than about 200 picomolar. D In certain embodiments, antibodies that specifically bind to LIF have a K of less than about 100 picomolar. D In certain embodiments, the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriplatin, picoplatin, satraplatin, or a combination thereof. In certain embodiments, the cancer is resistant to treatment with a therapeutic amount of the LIF-binding polypeptide or platinum-based anti-cancer agent administered as monotherapy.

[0102] In certain embodiments, described herein is the use of a LIF-binding polypeptide in combination with a platinum-based anti-neoplastic agent to treat glioblastoma multiforme in an individual. In certain embodiments, the LIF-binding polypeptide and the platinum-based anti-neoplastic agent are administered to the individual in separate formulations. In certain embodiments, the LIF-binding polypeptide and the platinum-based anti-neoplastic agent are administered to the individual in the same formulation. In certain embodiments, the LIF-binding polypeptide is administered to the individual before the platinum-based anti-neoplastic agent is administered to the individual. In certain embodiments, the platinum-based anti-neoplastic agent is administered to the individual before the LIF-binding polypeptide is administered to the individual. In certain embodiments, the platinum-based anti-neoplastic agent is administered to the individual simultaneously with the LIF-binding polypeptide. In certain embodiments, the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. In certain embodiments, the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. In certain embodiments, the antibody that specifically binds to LIF comprises at least one framework region derived from a human antibody framework region. In certain embodiments, an antibody that specifically binds to LIF is humanized. In certain embodiments, an antibody that specifically binds to LIF is deimmunized. In certain embodiments, an antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. In certain embodiments, an antibody that specifically binds to LIF is an IgG antibody. In certain embodiments, an antibody that specifically binds to LIF is a Fab, F(ab)2, single domain antibody, single chain variable fragment (scFv), or nanobody.In certain embodiments, an antibody that specifically binds to LIF comprises: (a) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; (b) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; (c) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; (d) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; (e) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and (f) an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13. In certain embodiments, an antibody that specifically binds LIF comprises (a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and (b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45-48. In certain embodiments, the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO:42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO:46.In certain embodiments, an antibody that specifically binds to LIF comprises (a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60 or 67; and (b) an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. In certain embodiments, an antibody that specifically binds to LIF has a K of less than about 200 picomolar. D In certain embodiments, antibodies that specifically bind to LIF have a K of less than about 100 picomolar. D In certain embodiments, the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriplatin, picoplatin, satraplatin, or a combination thereof. In certain embodiments, the cancer is resistant to treatment with a therapeutic amount of the LIF-binding polypeptide or platinum-based anti-cancer agent administered as monotherapy.

[0103] As used herein, "treating" or "treatment" refers to an intervention in a pathological condition intended to produce one or more beneficial effects. For cancer / tumors, treatment includes methods intended to result in or ensure stable disease, a partial response, a complete remission, increased progression-free survival, increased overall survival, tumor shrinkage, slowed tumor growth, halted tumor growth, or the prevention or reduction of metastasis. In certain cases, the treatment methods described herein can be used as maintenance after successful treatment or to prevent recurrence or metastasis of a particular tumor or cancer. It will be understood that not all individuals will respond to a given administration of a therapeutic antibody to the same extent or at all, but that these individuals are considered to be treated even if no response is detected.

[0104] Exemplary Embodiments Exemplary embodiments include: 1. Use of a leukemia inhibitory factor (LIF)-binding polypeptide in combination with a platinum-based antineoplastic agent to treat cancer in an individual; 2. Use of embodiment 1, wherein the LIF-binding polypeptide and the platinum-based antineoplastic agent are administered to the individual in separate formulations; 3. Use of embodiment 1, wherein the LIF-binding polypeptide and the platinum-based antineoplastic agent are administered to the individual in the same formulation; 4. Use of embodiment 1 or 2, wherein the LIF-binding polypeptide is administered to the individual before the platinum-based antineoplastic agent is administered to the individual; 5. Use of embodiment 1 or 2, wherein the platinum-based antineoplastic agent is administered to the individual before the LIF-binding polypeptide is administered to the individual; 6. Use of any one of embodiments 1 to 3, wherein the platinum-based antineoplastic agent is administered to the individual at the same time as the LIF-binding polypeptide is administered to the individual; and 7. Use of any one of embodiments 1 to 6, wherein the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. 8. The use of embodiment 7, wherein the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. 9. The use of embodiment 8, wherein the antibody that specifically binds to LIF comprises at least one framework region derived from a human antibody framework region. 10. The use of embodiment 8, wherein the antibody that specifically binds to LIF is humanized. 11. The use of embodiment 8, wherein the antibody that specifically binds to LIF is deimmunized. 12. The use of embodiment 8, wherein the antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. 13. The use of embodiment 8, wherein the antibody that specifically binds to LIF is an IgG antibody. 14. The use of embodiment 8, wherein the antibody that specifically binds to LIF is a Fab, F(ab)2, single domain antibody, single chain variable fragment (scFv), or nanobody.15. Use according to any one of embodiments 8 to 14, wherein the antibody that specifically binds to LIF comprises: a) immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; b) immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; c) immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; d) immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; e) immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and f) immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13. 16. The use of any one of embodiments 8 to 15, wherein the antibody that specifically binds to LIF comprises: (a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and (b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45 to 48. 17. The use of embodiment 16, wherein the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO: 46. 18. The use of embodiment 17, wherein the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO: 46.19. The use of any one of embodiments 8 to 18, wherein the antibody that specifically binds to LIF comprises: a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57 to 60 or 67; and b) an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61 to 64. 20. The antibody that specifically binds to LIF has a K of less than about 200 picomolar. D 21. The use of any one of embodiments 8 to 19, wherein the antibody that specifically binds to LIF binds at a K of less than about 100 picomolar. D22. The use of any one of embodiments 10 to 21, wherein the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriptin, picoplatin, satraplatin, or a combination thereof. 23. The use of embodiment 22, wherein the platinum-based anti-cancer agent is cisplatin. 24. The use of any one of embodiments 1 to 23, wherein the cancer comprises advanced solid tumors, glioblastoma, gastric cancer, skin cancer, prostate cancer, pancreatic cancer, breast cancer, testicular cancer, thyroid cancer, head and neck cancer, liver cancer, kidney cancer, esophageal cancer, ovarian cancer, colon cancer, lung cancer, lymphoma, or soft tissue cancer. 25. The use of embodiment 24, wherein the cancer comprises non-small cell lung cancer, ovarian epithelial cancer, or pancreatic adenocarcinoma. 26. The use of any one of embodiments 1-25, wherein the cancer is resistant to treatment with a therapeutic amount of a LIF-binding polypeptide or a platinum-based antineoplastic agent administered as monotherapy. 27. A method of treating an individual with cancer, comprising administering to the individual with cancer an effective amount of a combination of (a) a leukemia inhibitory factor (LIF)-binding polypeptide and (b) a platinum-based antineoplastic agent. 28. The method of embodiment 27, comprising administering to the individual with cancer an effective amount of a LIF-binding polypeptide. 29. The method of embodiment 27, comprising administering to the individual with cancer an effective amount of a platinum-based antineoplastic agent. 30. The method of any one of embodiments 27-29, wherein the LIF-binding polypeptide comprises a fragment of an immunoglobulin variable region or an immunoglobulin heavy chain constant region. 31. The method of any one of embodiments 27-29, wherein the LIF-binding polypeptide comprises an antibody that specifically binds to LIF. 32. The method of embodiment 31, wherein the antibody that specifically binds to LIF comprises at least one framework region derived from a human antibody framework region. 33. The method of embodiment 31, wherein the antibody that specifically binds to LIF is humanized. 34. The method of embodiment 31, wherein the antibody that specifically binds to LIF is deimmunized. 35. The method of embodiment 31, wherein the antibody that specifically binds to LIF comprises two immunoglobulin heavy chains and two immunoglobulin light chains. 36. The method of embodiment 31, wherein the antibody that specifically binds to LIF is an IgG antibody.37. The method of embodiment 31, wherein the antibody that specifically binds to LIF is a Fab, F(ab)2, single domain antibody, single chain variable fragment (scFv), or nanobody. 38. The method of any one of embodiments 31 to 37, wherein the antibody that specifically binds to LIF comprises: a) immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; b) immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; c) immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; d) immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; e) immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and f) immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13. 39. The method of any one of embodiments 31 to 37, wherein the antibody that specifically binds to LIF comprises: a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 41, 42, 44, or 66; and b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 45 to 48. 40. The method of embodiment 39, wherein the VH sequence is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 46. 41. The method of embodiment 40, wherein the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO: 46.42. The method of any one of embodiments 31-37, wherein the antibody that specifically binds to LIF comprises: a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 57-60 or 67; and b) an immunoglobulin light chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in any one of SEQ ID NOs: 61-64. 43. The method of any one of embodiments 31-42, wherein the antibody that specifically binds to LIF binds with a KD of less than about 200 picomolar. 44. The method of any one of embodiments 31-42, wherein the antibody that specifically binds to LIF binds with a KD of less than about 100 picomolar. 45. The method of any one of embodiments 27-44, wherein the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriptin, picoplatin, satraplatin, or a combination thereof. 46. The method of embodiment 45, wherein the platinum-based anti-cancer agent is cisplatin. 47. The method of any one of embodiments 27-46, wherein the cancer comprises advanced solid tumors, glioblastoma, gastric cancer, skin cancer, prostate cancer, pancreatic cancer, breast cancer, testicular cancer, thyroid cancer, head and neck cancer, liver cancer, kidney cancer, esophageal cancer, ovarian cancer, colon cancer, lung cancer, lymphoma, or soft tissue cancer. 48. The method of embodiment 47, wherein the cancer comprises non-small cell lung cancer, ovarian epithelial cancer, or pancreatic adenocarcinoma. 49. The method of any one of embodiments 27-48, wherein the cancer is resistant to treatment with a therapeutic amount of an inhibitor of a LIF-binding polypeptide. 50. The method of any one of embodiments 27-48, wherein the cancer is resistant to treatment with a therapeutic amount of a platinum-based antineoplastic agent. 51. The method of any one of embodiments 27-50, wherein the leukemia inhibitory factor (LIF)-binding polypeptide and the platinum-based antineoplastic agent are administered separately. 52. The method of any one of embodiments 27-50, wherein the LIF-binding polypeptide and the platinum-based antineoplastic agent are administered simultaneously. 53. The method of any one of embodiments 27-50, wherein the LIF-binding polypeptide and the platinum-based antineoplastic agent are administered in a single composition. 54. Use of an antibody that specifically binds leukemia inhibitory factor (LIF) in combination with a platinum-based antineoplastic agent to treat cancer in an individual.Here, the LIF-binding antibody is selected from the group consisting of: a) immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; b) immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; c) immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; d) immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10; e) immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and f) immunoglobulin light chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13. 55. A method of treating an individual with cancer, comprising: a) selecting from: i. an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 3; ii. an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 4 or 5; iii. an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 6 to 8; iv. an immunoglobulin heavy chain complementarity determining region 4 (VH-CDR5) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9 or 10. v. an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and vi. an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13; and b) an effective amount of a platinum-based antineoplastic agent, in combination with an antibody that specifically binds to leukemia inhibitory factor (LIF), the antibody comprising: an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11 or 12; and vi. an immunoglobulin light chain complementarity determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13; and b) an effective amount of a platinum-based antineoplastic agent, to an individual having cancer. 56. The method of embodiment 55, comprising administering an effective amount of a LIF-binding polypeptide to an individual having cancer. 57. The method of embodiment 55, comprising administering an effective amount of a platinum-based antineoplastic agent to an individual having cancer. [Example]

[0105] The following examples are representative of embodiments of the compositions and methods described herein and are not intended to be limiting in any way.

[0106] Example 1 - Production of rat antibodies specific for LIF A cDNA encoding amino acids 23-202 of human LIF was cloned into an expression plasmid (Aldevron GmbH, Freiburg, Germany). Groups of laboratory rats (Wistar) were immunized by intradermal application of DNA-coated gold particles using a handheld device for particle bombardment (gene gun). Cell surface expression on transiently transfected HEK cells was confirmed using an anti-tag antibody that recognizes the tag added to the N-terminus of the LIF protein. After a series of immunizations, serum samples were collected and tested by flow cytometry against HEK cells transiently transfected with the aforementioned expression plasmid. Antibody-producing cells were isolated and fused with mouse myeloma cells (Ag8) according to standard procedures. Hybridomas producing LIF-specific antibodies were identified by screening using flow cytometry analysis as described above. Cell pellets of positive hybridoma cells were prepared using an RNA protection agent (RNAlater, cat. #AM7020 by ThermoFisher Scientific) and further processed for sequencing of the variable domains of the antibodies.

[0107] Example 2 - Production of mouse antibodies specific for LIF A cDNA encoding amino acids 23-202 of human LIF was cloned into an expression plasmid (Aldevron GmbH, Freiburg, Germany). Groups of laboratory mice (NMRI) were immunized by intradermal application of DNA-coated gold particles using a handheld device for microparticle bombardment (gene gun). Cell surface expression on transiently transfected HEK cells was confirmed using an anti-tag antibody that recognizes the tag added to the N-terminus of the LIF protein. After a series of immunizations, serum samples were collected and tested by flow cytometry against HEK cells transiently transfected with the aforementioned expression plasmid. Antibody-producing cells were isolated and fused with mouse myeloma cells (Ag8) according to standard procedures. Hybridomas producing LIF-specific antibodies were identified by screening using flow cytometry analysis as described above. Cell pellets of positive hybridoma cells were prepared using an RNA protection agent (RNAlater, cat. #AM7020 by ThermoFisher Scientific) and further processed for sequencing of the variable domains of the antibodies.

[0108] Example 3 - Humanization of a rat antibody specific for LIF One clone from the rat immunization (5D8) was selected for subsequent humanization. Humanization was performed using standard CDR-grafting methods. Heavy and light chain regions were cloned from the 5D8 hybridoma using standard molecular cloning techniques and sequenced by the Sanger method. BLAST searches were then performed against human heavy and light chain variable sequences, and four sequences from each were selected as acceptor frameworks for humanization. These acceptor frameworks were deimmunized to remove T cell response epitopes. The heavy and light chain CDR1, CDR2, and CDR3 of 5D8 were cloned into four different heavy chain acceptor frameworks (H1 to H4) and four different light chain frameworks (L1 to L4). All 16 different antibodies were then tested for expression in CHO-S cells (Selexis); inhibition of LIF-induced STAT3 phosphorylation; and binding affinity by surface plasmon resonance (SPR). These experiments are summarized below.

[0109] [Table 1]

[0110] The expression performance of the transfected cells was evaluated by culturing 3 × 10 cells in Erlenmeyer flasks. 5 The results were compared after 10 days of cell culture in fed-batch cultures (200 mL culture volume, 200 cells / mL seeding). At this point, cells were harvested and secreted antibodies were purified using a Protein A column and then quantified. All humanized antibodies were expressed except for those using the H3 heavy chain (SEQ ID NO: 43).

[0111] Inhibition of LIF-induced STAT3 phosphorylation at tyrosine 705 was determined by Western blot. U251 glioma cells were plated in six-well plates at a density of 100,000 cells / well. Cells were cultured in complete medium for 24 hours before any treatment, and then serum-starved for 8 hours. Cells were then treated overnight with the indicated antibodies at a concentration of 10 μg / ml. After treatment, proteins were obtained and quantified (BCA-protein quantification, Thermo Fisher Scientific) in radioimmunoprecipitation assay (RIPA) lysis buffer containing phosphatase and protease inhibitors and used for Western blot analysis. For Western blots, membranes were blocked in 5% nonfat dry milk-TBST for 1 hour and incubated with primary antibodies overnight (p-STAT3, catalog #9145, Cell Signaling or STAT3, catalog #9132, Cell Signaling) or for 30 minutes (β-actin-peroxidase, catalog #A3854, Sigma-Aldrich). The membranes were then washed with TBST, incubated with secondary antibodies, and washed again. Proteins were detected by chemiluminescence (SuperSignal Substrate, catalog #34076, Thermo Fisher Scientific). The results are shown in Figure 1. The darker the pSTAT3 band, the less inhibition was present. Inhibition was high in lanes labeled 5D8 (non-humanized rat), A (H0L0), C (H1L2), D (H1L3), and G (H2L2); inhibition was moderate in H (H2L3), O (H4L2), and P (H4L3); inhibition was absent in B (H1L1), E (H1L4), F (H2L1), I (H2L4), N (H4L1), and Q (H4L4).

[0112] Antibodies that exhibited inhibition of LIF-induced STAT3 phosphorylation were then analyzed by SPR to determine binding affinity. Briefly, the binding of A (H0L0), C (H1L2), D (H1L3), and G (H2L2), H (H2L3), and O (H4L2) humanized antibodies to amine-coupled hLIF was monitored using a Biacore™ 2002 instrument. The rate constants and affinities were determined by mathematical sensorgram fitting (Langmuir interaction model [A + B = AB]) of all sensorgrams generated on all sensor chip surfaces at six ligand concentrations. The best-fit curve (least chi-squared) for each concentration was used to calculate the rate constants and affinities. See Table 1.

[0113] Because the experimental procedure used bivalent antibodies as analytes, the best-fit sensorgrams were also analyzed based on a bivalent analyte fitting model [A + B = AB; AB + B = AB2] to gain more insight into the target binding mechanism of the humanized antibodies. The relative affinity ranking of the mAb samples was confirmed by sensorgram kinetic analysis using the bivalent fitting model [A + B = AB; AB + B = AB2].

[0114] Due to its high binding affinity and high yield from batch cultures, humanized 5D8 containing H2 and L2 was selected for more in-depth analysis.

[0115] Example 4 - Humanization of clone 5D8 improves binding to LIF The H2L2 clone (h5D8) was selected for further analysis, and its binding was compared by SPR to that of the parent rat 5D8 (r5D8) and the mouse clone 1B2. The 1B2 antibody is a previously described mouse anti-LIF antibody previously deposited with Deutsche Sammlung von Mikroorganismen and Zellkulturen GmbH (DSM ACC3054) and was included for comparative purposes. Recombinant human LIF purified from Escherichia coli (E. coli) and HEK-293 cells, respectively, was used as the ligand. LIF from human or E. coli sources was covalently coupled to the surface of a Biacore optical sensor chip using amine coupling chemistry, and binding affinities were calculated from rate constants.

[0116] Materials and Methods Human LIF from Escherichia coli (E. coli) was obtained from Millipore (reference LIF 1010); human LIF from HEK-293 cells was obtained from ACRO Biosystems (reference LIF-H521b). LIF was coupled to the sensor chip using a Biacore Amine Coupling Kit (BR-1000-50; GE-Healthcare, Uppsala). Samples were run on a Biacore™ 2002 instrument using a CM5 optical sensor chip (BR-1000-12; GE-Healthcare, Uppsala). Biacore HBS-EP buffer was used during machine operation (BR-1001-88; GE-Healthcare, Uppsala). Kinetic analysis of binding sensorgrams was performed using BIAevaluation 4.1 software. The rate constants and affinities were determined by mathematical sensorgram fitting (Langmuir interaction model [A + B = AB]) of all sensorgrams generated on all sensor chip surfaces at increasing analyte concentrations. To obtain an estimate of the bivalent contribution (e.g., avidity contribution) to the determined Langmuir antibody-target affinity, the sensorgrams were also analyzed based on a bivalent analyte sensorgram fitting model [A + B = AB; AB + B = AB2], including principal component analysis. The best-fit curve (least chi-squared) for each concentration was used to calculate the rate constants and affinities. A summary of these affinity experiments is shown in Table 2 (human LIF produced in E. coli) and Table 3 (human LIF produced in HEK293 cells).

[0117] [Table 2]

[0118] [Table 3]

[0119] From this series of experiments, a Langmuir 1:1 sensorgram fitting model indicates that the humanized 5D8 (h5D8) antibody bound to human LIF with approximately 10-25 times higher affinity than murine 1B2 and r5D8.

[0120] Next, the h5D8 antibody was tested against LIF from multiple species by SPR. h5D8 SPR binding kinetics were performed on recombinant LIF samples from different species and expression systems: human LIF (E. coli, HEK293 cells); mouse LIF (E. coli, CHO cells); rat LIF (E. coli); and cynomolgus monkey LIF (yeast, HEK293 cells).

[0121] Materials and Methods The h5D8 antibody was immobilized on the sensor chip surface by noncovalent, Fc-specific capture. Recombinant Ig(Fc)-specific Staphylococcus aureus (S. aureus) protein A / G was used as the capture agent, allowing for spatially uniform and flexible presentation of anti-LIF antibodies to LIF analytes. The sources of LIF analytes were as follows: human LIF (from E. coli; Millipore reference LIF 1050); human LIF (from HEK cells; ACRO Biosystems LIF-H521); mouse LIF (from E. coli; Millipore Cat. No. NF-LIF2010); mouse LIF (from CHO cells; Reprokine Cat. No. RCP09056); monkey LIF (yeast Kingfisher Biotech Cat. No. RP1074Y); and monkey LIF produced in HEK-293 cells. All h5D8 antibodies exhibited binding to LIF from several species. A summary of this affinity experiment is shown in Table 4.

[0122] [Table 4]

[0123] Example 5 - Humanized clone 5D8 inhibits LIF-induced phosphorylation of STAT3 in vitro To determine the biological activity of h5D8, the humanized and parental forms were tested in a cell culture model of LIF activation. Figure 2A shows that the humanized clone exhibited increased inhibition of STAT3 phosphorylation (Tyr 705) when glioma cell lines were incubated with human LIF. Figure 2B shows an experiment using the same procedure as in Figure 2A, but performed again with different dilutions of h5D8 antibody.

[0124] method U251 glioma cells were plated in 6-well plates at a density of 150,000 cells / well. Cells were cultured in complete medium for 24 hours before any treatment. Cells were then treated overnight with r5D8 or h5D8 anti-LIF antibodies at a concentration of 10 μg / ml, or not (control cells).

[0125] After treatment, proteins were obtained and quantified in radioimmunoprecipitation assay (RIPA) lysis buffer containing phosphatase and protease inhibitors (BCA-Protein Quantitation, Thermo Fisher Scientific) and used for Western blotting. For Western blotting, membranes were blocked in 5% nonfat dry milk-TBST for 1 hour and incubated with primary antibodies overnight (p-STAT3, catalog #9145, Cell Signaling or STAT3, catalog #9132, Cell Signaling) or for 30 minutes (β-actin-peroxidase, catalog #A3854, Sigma-Aldrich). The membranes were then washed with TBST, incubated with secondary antibodies if necessary, and washed again. Proteins were detected by chemiluminescence (SuperSignal Substrate, catalog #34076, Thermo Fisher Scientific).

[0126] Example 6 - IC of h5D8 antibody treatment on endogenous levels of LIF in U-251 cells 50 value In U-251 cells, h5D8 showed a low IC of 490 picomolar for biological inhibition under serum-starved conditions (Figure 3A). 50 was also determined. See representative results Figures 3A and 3B and Table 5.

[0127] [Table 5]

[0128] method U-251 cells were plated at 600,000 cells per 6 cm plate (per condition). Cells were treated with h5D8 at the corresponding concentrations (serial dilution (titration)) overnight at 37°C under serum starvation (0.1% FBS). As a positive control for pSTAT3, recombinant LIF (R&D #7734-LF / CF) was used to stimulate cells at 1.79 nM for 10 minutes at 37°C. As a negative control for pSTAT3, JAK I inhibitor (Calbiochem #420099) was used at 1 μM for 30 minutes at 37°C. The cells were then harvested on ice for lysates and the detectable protein levels were measured by MSD Meso Sector S600 according to the Meso Scale Discovery Multi-Spot Assay System Total STAT3 (Cat# K150SND-2) and Phospho-STAT3 (Tyr705) (Cat# K150SVD-2) kit protocols.

[0129] Example 7 - Additional antibodies that specifically bind to human LIF Other rat antibody clones (10G7 and 6B5) that specifically bind to human LIF were identified and their binding characteristics are summarized in Table 6 below (clone 1B2 serves as a comparison).

[0130] method Real-time binding kinetics analysis was performed using recombinant LIF target proteins [human LIF (E. coli); Millipore Cat. No. LIF 1010 and human LIF (HEK293 cells); ACRO Biosystems Cat. No. LIF-H521b] immobilized on the surface of a CM5 optical sensor chip.

[0131] The rate constants and affinities were obtained by mathematical sensorgram fitting using a Langmuir 1:1 binding model, applying a global fitting (simultaneous fitting of a set of sensorgrams) and a single curve fitting algorithm. The adequacy of the global fit was assessed by k obs It was evaluated by analysis.

[0132] [Table 6]

[0133] Example 8 - Additional anti-LIF antibodies inhibit LIF-induced phosphorylation of STAT3 in vitro Additional clones were tested for their ability to inhibit LIF-induced phosphorylation of STAT3 in cell culture. As shown in Figure 4, clones 10G7 and r5D8, previously detailed, exhibited greater inhibition of LIF-induced STAT3 phosphorylation compared to clone 1B2. Anti-LIF polyclonal antiserum (pos.) was included as a positive control. 6B5 exhibited no inhibition, which may be explained by the possible absence of 6B5 binding to the nonglycosylated LIF used in this experiment.

[0134] method Patient-derived glioma cells were plated at a density of 150,000 cells / well in six-well plates. Prior to any treatment, cells were cultured for 24 hours in GBM medium (Life Technologies) consisting of Neurobasal medium supplemented with B27 (Life Technologies), penicillin / streptomycin, and growth factors (20 ng / ml EGF and 20 ng / ml FGF-2 [PeproTech]). The following day, cells were treated with or without recombinant LIF produced in Escherichia coli (E. coli) or a mixture of recombinant LIF plus the indicated antibodies for 15 minutes (10 μg / ml final concentration for antibodies and 20 ng / ml recombinant LIF). After treatment, proteins were obtained and quantified (BCA-protein quantification, Thermo Fisher Scientific) in radioimmunoprecipitation assay (RIPA) lysis buffer containing phosphatase and protease inhibitors and used for Western blot analysis. For Western blots, membranes were blocked in 5% nonfat dry milk-TBST for 1 hour and incubated with primary antibodies overnight (p-STAT3, catalog #9145, Cell Signaling) or for 30 minutes (β-actin-peroxidase, catalog #A3854, Sigma-Aldrich). The membranes were then washed with TBST, incubated with secondary antibodies if necessary, and washed again. Proteins were detected by chemiluminescence (SuperSignal Substrate, catalog #34076, Thermo Fisher Scientific).

[0135] Example 9 - LIF is highly overexpressed across multiple tumor types Immunohistochemistry was performed on multiple human tumor types to determine the extent of LIF expression. As shown in Figure 5, LIF is highly expressed in glioblastoma multiforme (GBM), non-small cell lung cancer (NSCLC), ovarian cancer, colorectal cancer (CRC), and pancreatic cancer.

[0136] Example 10 - Humanized clone h5D8 inhibits tumor growth in a mouse model of non-small cell lung cancer To determine the ability of the humanized 5D8 clone to inhibit LIF-positive cancer in vivo, the antibody was tested in a mouse model of non-small cell lung cancer (NSCLC). Figure 6A shows reduced tumor growth in mice treated with this antibody compared to the vehicle negative control. Figure 6B shows data obtained using the r5D8 version.

[0137] method The mouse non-small cell lung cancer (NSCLC) cell line KLN205, which has high LIF levels, was stably infected with a lentivirus expressing the firefly luciferase gene for in vivo bioluminescence monitoring. To develop the mouse model, 5 × 10 5 KLN205 non-small cell lung cancer (NSCLC) cells were orthotopically implanted into the left lung of 8-week-old, immunocompetent, syngeneic DBA / 2 mice via intercostal puncture. Mice were treated with control vehicle or 15 mg / kg or 30 mg / kg of h5D8 antibody (intraperitoneally, twice weekly), and tumor growth was monitored by bioluminescence. For bioluminescence imaging, mice received an intraperitoneal injection of 0.2 mL of 15 mg / mL D-luciferin under 1-2% inhaled isoflurane anesthesia. Bioluminescence signals were monitored using an IVIS System 2000 Series (Xenogen Corp., Alameda, CA, USA), which consists of a high-sensitivity, cooled CCD camera. Using Living Image software (Xenogen Corp.), imaging data were gridded and the total bioluminescence signal within each enclosed region was integrated. Data were analyzed using the total photon flux emission (photons / second) within the region of interest (ROI). The results demonstrate that treatment with h5D8 antibody promotes tumor regression. Data are presented as mean ± SEM.

[0138] Example 11 - h5D8 inhibits tumor growth in a mouse model of glioblastoma multiforme In an orthotopic GBM tumor model using the luciferase-expressing human cell line U251, r5D8 significantly reduced tumor volume in mice administered 300 μg r5D8 and h5D8 twice weekly via intraperitoneal (IP) injection. The results of this study are shown in Figure 7A (quantified on day 26 after treatment). This experiment was also performed using humanized h5D8 mice treated with 200 μg or 300 μg h5D8, demonstrating a statistically significant reduction in tumor volume 7 days after treatment. Figure 7B shows data from mice inoculated with luciferase-expressing human U251 GBM cells and subsequently treated twice weekly with 100, 200, or 300 μg h5D8 or vehicle. Tumor size was determined by bioluminescence (Xenogen IVIS Spectrum) on day 7. Graphs show individual tumor measurements with horizontal bars indicating mean ± SEM. Statistical significance was calculated using the unpaired nonparametric Mann-Whitney U test.

[0139] method U251 cells stably expressing luciferase were harvested, washed in PBS, centrifuged at 400 g for 5 minutes, resuspended in PBS, and counted using an automated cell counter (Countess, Invitrogen). Cells were kept on ice to maintain optimal viability. Mice were anesthetized with intraperitoneal administration of ketamine (Ketolar50®) / xylacine (Rompun®) (75 mg / kg and 10 mg / kg, respectively). Each mouse was carefully placed in a stereotaxic device and immobilized. Hair was removed from the head using depilatory cream, and the scalp was cut with a scalpel to expose the skull. A small incision was carefully made using a trephine at coordinates 1.8 mm lateral and 1 mm anterior to lambda. 5 μL of cells were inoculated into the right striatum at a depth of 2.5 mm using a Hamilton 30G syringe. The head incision was closed using Hystoacryl tissue adhesive (Braun) and the mice were injected with the subcutaneous analgesic meloxicam (Metacam®) (1 mg / kg). The final number of cells implanted in each mouse was 3 x 10 5 There were 100 pieces.

[0140] Mice were treated twice weekly with h5D8 administered intraperitoneally. Treatment began immediately after tumor cell inoculation, on day 0. Mice received a total of two doses of h5D8 or vehicle control.

[0141] Body weight and tumor volume: Body weight was measured twice weekly, and tumor growth was quantified by bioluminescence (Xenogen IVIS Spectrum) on day 7. To quantify bioluminescence activity in vivo, mice were anesthetized with isofluorane and injected intraperitoneally with luciferin substrate (PerkinElmer) (167 μg / kg).

[0142] Tumor size, determined by bioluminescence (Xenogen IVIS Spectrum), was assessed on day 7. Mean ± SEM values ​​were calculated for individual tumor measurements and for each treatment group. Statistical significance was determined by the unpaired, nonparametric Mann-Whitney U test.

[0143] Example 12 - h5D8 inhibits tumor growth in a mouse model of ovarian cancer The efficacy of r5D8 was evaluated in two other syngeneic tumor models. In the ovarian orthotopic tumor model ID8, IP administration of 300 μg r5D8 (twice weekly) significantly inhibited tumor growth as measured by abdominal volume (Figures 8A and 8B). The results in Figure 8C show that h5D8 also reduced tumor volume at doses of 200 μg or higher.

[0144] method ID8 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) (Gibco, Invitrogen) supplemented with 10% fetal bovine serum (FBS) (Gibco, Invitrogen), 40 U / mL penicillin and 40 μg / mL streptomycin (PenStrep) (Gibco, Invitrogen), and 0.25 μg / mL plasmocin (Invivogen).

[0145] ID8 cells were harvested, washed in PBS, centrifuged at 400 g for 5 min, and resuspended in PBS. To maintain optimal viability, cells were kept on ice, and 200 μL of the cell suspension was injected intraperitoneally using a 27G needle. The final number of cells implanted in mice was 5 × 10 6 There were 100 pieces.

[0146] Mice were treated twice weekly with h5D8 administered i.p. at different doses as indicated. Body weight was measured twice weekly, and tumor progression was monitored by measuring abdominal circumference using a vernier caliper (Fisher Scientific).

[0147] Example 13 - r5D8 inhibits tumor growth in a mouse model of colorectal cancer In mice bearing subcutaneous colon CT26 tumors, r5D8 (administered IP at 300 μg twice weekly) significantly inhibited tumor growth (FIGS. 9A and 9B).

[0148] method CT26 cells were cultured in Roswell Park Memorial Institute medium (RPMI [Gibco, Invitrogen]) supplemented with 10% fetal bovine serum (FBS), 40 U / mL penicillin and 40 μg / mL streptomycin (PenStrep), and 0.25 μg / mL plasmocin.

[0149] CT26 cells (8 x 10 5 ) were trypsinized, rinsed with PBS, centrifuged at 400 g for 5 minutes, and resuspended in 100 μL PBS. Cells were kept on ice to avoid cell death. CT26 cells were administered to mice via subcutaneous injection using a 27G needle.

[0150] Mice were administered 300 μg r5D8, or vehicle control, via intraperitoneal injection (IP) twice weekly starting on day 3 after CT26 cell implantation.

[0151] Body weight and tumor volume were measured three times a week. Tumor volume was measured using a vernier caliper (Fisher Scientific).

[0152] Example 14 - r5D8 reduces inflammatory infiltrates in tumor models In a U251 GBM orthotopic transplant model, expression of CCL22, a marker of M2-polarized macrophages, was significantly reduced in tumors treated with r5D8, as shown in Figure 10A. This finding was also confirmed in a physiologically relevant organotypic explant culture model using h5D8, where three patient samples showed a significant reduction in CCL22 and CD206 (MRC1) expression (also a marker of M2 macrophages) after treatment, as shown in Figure 10B (compare top left (control) with bottom right (treated) for both MRC1 and CCL22). Furthermore, r5D8 also reduced CCL22 expression in syngeneic ID8 (Figure 10C) and CT26 (Figure 10D) tumors in immunocompetent mice. + H5D8 treatment also programmed macrophages to an immunostimulatory phenotype in a syngeneic CT26 tumor model (Figure 10E). H5D8 treatment increased macrophages with an M1 phenotype, as indicated by an increased CD206-negative / MHCII-positive ratio, and decreased macrophages with an M2 phenotype, as indicated by a decreased CD206-positive / MHCII-negative ratio. Figure 10F shows gene expression data from monocytes cultured in conditioned medium from U251 cells with LIF knockdown. MRC1, CCL2, CCL1, and CTSK (represented by triangles) all showed significant decreases in expression.

[0153] Example 15 - r5D8 Increases Non-Myeloid Effector Cells To explore further immune mechanisms, we evaluated the effect of r5D8 on T cells and other non-myeloid immune effector cells within the tumor microenvironment. In the ovarian orthotopic ID8 syngeneic transplant model, r5D8 treatment significantly increased intratumoral NK cells and total and activated CD4+ cells, as shown in Figure 11A. + and CD8 +Similarly, in the colon isograft CT26 tumor model, r5D8 increased intratumoral NK cells and CD4 + and CD8 + Increases T cells and CD4 + CD25 + FoxP3 + There was a tendency to decrease T-reg cells. + CD25 + FoxP3 + A trend toward a decrease in T-reg cells was also observed in the syngeneic and orthotopic KLN205 tumor model after r5D8 treatment, as shown in Figure 11C. Consistent with the requirement for T cells to mediate efficacy, the CD4 + and CD8 + T cell depletion inhibited the antitumor efficacy of r5D8, as shown in FIG.

[0154] Methods for T cell depletion CT26 cells were cultured in RPMI culture medium (Gibco, Invitrogen) supplemented with 10% fetal bovine serum (FBS [Gibco, Invitrogen]), 40 U / mL penicillin and 40 μg / mL streptomycin (PenStrep [Gibco, Invitrogen]), and 0.25 μg / mL plasmocin. CT26 cells (5 × 10 5 ) were collected, rinsed with PBS, centrifuged at 400 g for 5 minutes, and resuspended in 100 μL PBS. Cells were kept on ice to avoid cell death. CT26 cells were administered to mice via subcutaneous injection into both flanks using a 27G syringe. Mice were treated twice weekly with r5D8 administered intraperitoneally as directed in the study design. Vehicle control (PBS), rat r5D8, and / or anti-CD4 and anti-CD8 were administered to mice via intraperitoneal injection (IP) twice weekly as described in the study design. All antibody treatments were administered simultaneously.

[0155] Example 16 - Crystal structure of h5D8 in complex with human LIF The crystal structure of h5D8 was analyzed to 3.1 Å resolution to determine the epitope on LIF to which h5D8 binds and to determine the residues of h5D8 involved in binding. The co-crystal structure revealed that the N-terminal loop of LIF is centrally located between the light and heavy chain variable regions of h5D8 (Figure 13A). Furthermore, h5D8 interacts with residues on helices A and C of LIF, thereby forming a discontinuous and conformational epitope. Binding is driven by several salt bridges, H-bonds, and van der Waals interactions (Table 7, Figure 13B). The h5D8 epitope on LIF spans the region of interaction with gp130. See Boulanger, MJ, Bankovich, AJ, Kortemme, T., Baker, D. & Garcia, KC Convergent mechanisms for recognition of divergent cytokines by the shared signaling receptor gp130. Molecular cell 12, 577-589 (2003). These results are summarized in Table 7 below and depicted in Figure 13.

[0156] [Table 7]

[0157] method LIF was generated from HEK 293S (Gnt I - / -() cells and purified using Ni-NTA affinity chromatography followed by gel filtration chromatography (in 20 mM Tris pH 8.0 and 150 mM NaCl). Recombinant h5D8 Fab was transiently expressed in HEK 293F cells and purified using KappaSelect affinity chromatography followed by cation exchange chromatography. Purified h5D8 Fab and LIF were mixed at a molar ratio of 1:2.5 and incubated at room temperature for 30 minutes, followed by deglycosylation using EndoH. The complex was then purified using gel filtration chromatography. The complex was concentrated to 20 mg / mL and prepared for crystallization trials using a tesparse matrix screen. Crystals were formed at 4°C in a solution containing 19% (v / v) isopropanol, 19% (w / v) PEG 4000, 5% (v / v) glycerol, and 0.095 M sodium citrate (pH 5.6). The crystals were diffracted to 3.1 Å resolution on the 08ID-1 beamline at the Canadian Light Source (CLS). Data were collected, processed, and scaled using XDS as described by Kabsch et al., Xds. Acta crystallographica. Section D, Biological crystallography 66, 125-132 (2010). The structure was determined by molecular replacement using Phaser crystallographic software as described by McCoy et al., Phaser crystallographic software. J Appl Crystallogr 40, 658-674 (2007). The structure was analyzed using the accepted R. work and R freeSeveral iterations of model building and refinement were performed using Coot and phenix.refine until convergence was achieved. See Emsley et al. Features and development of Coot. Acta crystallographica. Section D, Biological crystallography 66, 486-501 (2010); and Adams, et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta crystallographica. Section D, Biological crystallography 66, 213-221 (2010). Figures were generated with PyMOL (The PyMOL Molecular Graphics System, Version 2.0 Schroedinger, LLC).

[0158] Example 17 - h5D8 has high specificity for LIF The binding of h5D8 to other LIF family members was tested to determine binding specificity. Using Octet96 analysis, h5D8 binding to human LIF is approximately 100-fold stronger than its binding to oncostatin M (OSM), the IL-6 family member with the highest homology to LIF, when both proteins are produced in Escherichia coli (E. coli). When both proteins are produced in mammalian systems, h5D8 does not exhibit binding to OSM. The data are summarized in Table 8.

[0159] [Table 8]

[0160] method Octet Binding Experiment: Reagents were used and prepared as per the manufacturer's instructions. Basic Kinetics Experiments were performed using Octet Data Acquisition software version 9.0.0.26 as follows: Sensor / Program Settings: i) Equilibration (60 s); ii) Loading (15 s); iii) Baseline (60 s); iv) Binding (180 s); and v) Dissociation (600 s).

[0161] Octet affinity of h5D8 for cytokines: Basic kinetics experiments were performed using Octet Data Acquisition software version 9.0.0.26 as follows: Amine Reactive 2nd Generation Biosensors (AR2G) were hydrated in water for a minimum of 15 minutes. Amine coupling of h5D8 to the biosensor was performed using the Amine Coupling Second Generation Kit according to ForteBio Technical Note 26 (see references). Immersion steps were performed at 30°C and 1000 rpm as follows: i) 60 s equilibration in water; ii) 300 s activation in 20 mM ECD, 10 mM sulfo-NHS (in water); iii) 600 s immobilization of 10 μg / ml h5D8 in 10 mM sodium acetate (pH 6.0); iv) 300 s quench in 1 M ethanolamine (pH 8.5); v) 120 s baseline in water. Kinetic experiments were then performed at 30°C and 1000 rpm with the following soak and read steps: vi) a 60-second baseline in 1X kinetic buffer; vii) 180-second binding of appropriate serial dilutions of cytokines in 1X kinetic buffer; viii) 300-second dissociation in 1X kinetic buffer; and ix) three regeneration / neutralization cycles (5 seconds each for three cycles) alternating between 10 mM glycine (pH 2.0) and 1X kinetic buffer. After regeneration, the biosensor was reused for subsequent binding analyses.

[0162] Human recombinant LIF produced from mammalian cells was from ACROBiosystems (LIF-H521b); human recombinant OSM produced in mammalian cells was from R&D (8475-OM / CF); human recombinant OSM produced in Escherichia coli (E. coli) cells was from R&D (295-OM-050 / CF).

[0163] Example 18 - Crystal structure of h5D8 Fab Five crystal structures of h5D8 Fab were determined under a wide range of chemical conditions. The high resolution of these structures indicates that the conformations of the CDR residues are highly similar across different chemical environments, with only few degrees of freedom. A unique feature of this antibody is the presence of a non-canonical cysteine ​​at position 100 in the variable heavy chain region. Structural analysis indicates that the cysteine ​​is unpaired and highly inaccessible to solvent.

[0164] H5D8 Fab was obtained by papain digestion of its IgG, followed by purification using standard affinity, ion exchange, and chromatography techniques. Crystals were obtained using vapor diffusion, allowing the determination of five crystal structures with resolutions ranging from 1.65 Å to 2.0 Å. Despite crystallization conditions ranging across five different pH levels: 5.6, 6.0, 6.5, 7.5, and 8.5, all structures were solved within the same crystallographic space group and with similar unit cell dimensions (P212121, a ∼53.8 Å, b ∼66.5 Å, c ∼143.3 Å). Therefore, these crystal structures are unobstructed by crystal packing artifacts and allow comparison of the three-dimensional configuration of h5D8 Fab across a wide range of chemical conditions.

[0165] Electron density was observed for all complementarity-determining region (CDR) residues and subsequently modeled. Notably, LCDR1 and HCDR2 adopt an elongated conformation, which, together with the shallow LCDR3 and HCDR3 regions, forms the central binding groove of the paratope (Figure 14A). The five structures are highly similar across all residues, with root-mean-square deviations of all atoms ranging from 0.197 Å to 0.327 Å (Figure 14A). These results indicate that the conformation of the CDR residues is maintained in various chemical environments, including pH levels ranging from 5.6 to 8.5 and ionic strengths ranging from 150 mM to 1 M. Analysis of the electrostatic surface of the h5D8 paratope revealed that positively and negatively charged regions contribute equally to the hydrophilic character, with no prevalent hydrophobic patches. h5D8 possesses an unusual noncanonical cysteine ​​at the base of HCDR3 (Cys100). In all five structures, this free cysteine ​​is aligned and does not form any disulfide scrambles. Furthermore, it is not modified by the addition of Cys (cysteinylation) or glutathione (glutathiolation) and forms van der Waals interactions (3.5-4.3 Å distances) with the main and side chain atoms of Leu4, Phe27, Trp33, Met34, Glu102, and Leu105 of the heavy chain (Figure 14B). Finally, Cys100 is a predominantly buried structural residue that appears to be involved in mediating the conformation of CDR1 and HCDR3. Therefore, it is unlikely to be reactive with other cysteines, as observed by the uniform distribution of this region in our five crystal structures.

[0166] method H5D8-1 IgG was obtained from Catalent Biologics and formulated in 25 mM histidine, 6% sucrose, and 0.01% polysorbate 80 at pH 6.0. The formulated IgG was buffer-exchanged into PBS using a 10K MWCO concentrator (Millipore) on a large scale, followed by digestion with 1:100 micrograms of papain (Sigma) in PBS, 1.25 mM EDTA, and 10 mM cysteine ​​for 1 hour at 37°C. The papain-digested IgG was loaded onto a Protein A column (GE Healthcare) using an AKTA Start chromatography system (GE Healthcare). The Protein A flow-through containing the h5D8 Fab was collected and buffer-exchanged into 20 mM sodium acetate (pH 5.6) using a 10K MWCO concentrator (Millipore). The resulting sample was loaded onto a Mono S cation exchange column (GE Healthcare) using an AKTA Pure chromatography system (GE Healthcare). Elution with a 1 M potassium chloride gradient yielded a prominent h5D8 Fab peak, which was collected, concentrated, and purified to size homogeneity using a Superdex 200 Increase gel filtration column (GE Healthcare) in 20 mM Tris-HCl, 150 mM sodium chloride, pH 8.0. High purity of h5D8 Fab was confirmed by SDS-PAGE under reducing and non-reducing conditions.

[0167] The purified h5D8 Fab was concentrated to 25 mg / mL using a 10K MWCO concentrator (Millipore) and prepared for vapor diffusion crystallization experiments using low-density matrix 96-condition commercial screens JCSG TOP96 (Rigaku Reagents) and MCSG-1 (Anatrace) at 20 °C using an Oryx 4 dispenser (Douglas Instruments). The following five crystallization conditions were used: 1) 0.085 M sodium citrate, 25.5% (w / v) PEG 4000, 0.17 M ammonium acetate, 15% (v / v) glycerol (pH 5.6); 2) 0.1 M MES, 20% (w / v) PEG 6000, 1 M lithium chloride (pH 6.0); 3) 0.1 M MES, 20% (w / v) PEG 4000, 0.6 M sodium chloride (pH 6.5); 4) 0.085 M sodium HEPES, 17% (w / v) PEG 4000, 8.5% (v / v) 2-propanol, 15% (v / v) glycerol (pH 7.5); and 5) 0.08 M Tris, 24% (w / v) PEG. Crystals were obtained and collected after 4 days in 4000 MgCl, 0.16 M magnesium chloride, and 20% (v / v) glycerol (pH 8.5). Before flash freezing in liquid nitrogen, the mother liquor containing the crystals was supplemented with 5–15% (v / v) glycerol or 10% (v / v) ethylene glycol, as needed. Crystals were subjected to X-ray synchrotron radiation at the Advanced Photon Source, beamline 23-ID-D (Chicago, IL), and diffraction patterns were recorded with a Pilatus3 6M detector. Data were processed using XDS, and the structure was determined by molecular replacement using Phaser. Refinement was performed in PHENIX, with model building in Coot. Figures were generated using PyMOL. All software was accessed through SBGrid.

[0168] Example 19 - Mutation at Cysteine ​​100 of h5D8 maintains binding Analysis of h5D8 revealed a free cysteine ​​residue at position 100 (C100) in the variable region of the heavy chain. To characterize binding and affinity to human and mouse LIF, H5D8 variants were generated by substituting C100 with naturally occurring amino acids, respectively. Binding was characterized using ELISA and Octet assays. The results are summarized in Table 9. ELISA EC50 curves are shown in Figure 15 (Figure 15A human LIF and Figure 15B mouse LIF).

[0169] [Table 9]

[0170] method ELISA: Binding of h5D8 C100 variants to human and mouse LIF was determined by ELISA. Recombinant human or mouse LIF protein was coated onto Maxisorp 384-well plates at 1 μg / mL overnight at 4°C. Plates were blocked with 1× blocking buffer for 2 hours at room temperature. Serial dilutions (titrations) of each h5D8 C100 variant were added and allowed to bind for 1 hour at room temperature. Plates were washed three times with PBS + 0.05% Tween-20. HRP-conjugated anti-human IgG was added and allowed to bind for 30 minutes at room temperature. Plates were washed three times with PBS + 0.05% Tween-20 and developed using 1× TMB substrate. The reaction was stopped with 1 M HCl, and absorbance was measured at 450 nm. Figures and nonlinear regression analysis were performed using Graphpad Prism.

[0171] Octet RED96: The affinity of the h5D8 C100 variant for human and mouse LIF was determined by BLI using the Octet RED96 system. The h5D8 C100 variant was loaded onto an anti-human Fc biosensor at 7.5 μg / mL after a 30-second baseline in 1× kinetics buffer. A titration of serial dilutions of human or mouse LIF protein was allowed to bind to the loaded biosensor for 90 seconds and dissociate for 300 seconds in 1× kinetics buffer. KD was calculated by data analysis software using a 1:1 global fit model.

[0172] Example 20 - h5D8 blocks LIF binding to gp130 in vitro To determine whether h5D8 inhibits LIF binding to LIFR, a molecular binding assay using the Octet RED 96 platform was performed. H5D8 was loaded onto an AHC biosensor via anti-human Fc capture. The biosensor was then immersed in LIF, and binding was observed as expected (Figure 16A, middle third). The biosensor was then immersed in different concentrations of LIFR. Dose-dependent binding was observed (Figure 16A, right third). Control experiments demonstrated that this binding was LIF-specific (not shown) and not due to nonspecific interactions between LIFR and h5D8 or the biosensor.

[0173] To further characterize the binding of h5D8 to LIF, we performed a series of ELISA binding experiments. H5D8 and LIF were preincubated and then introduced to plates coated with recombinant human LIFR (hLIFR) or gp130. Lack of binding between the h5D8 / LIF complex and the coated substrate would indicate that h5D8 somehow interfered with LIF binding to its receptor. In addition, control antibodies that do not bind LIF (isotype control indicated by (-)) or that bind to LIF at a known binding site (B09 does not compete with either gp130 or LIFR for LIF binding; r5D8 is the rat parent form of h5D8) were also used. ELISA results demonstrated that the h5D8 / LIF complex (as well as the r5D8 / LIF complex) was capable of binding to hLIFR, indicating that these antibodies did not interfere with LIF / LIFR binding (Figure 16A). In contrast, the h5D8 / LIF complex (and r5D8 / LIF complex) was unable to bind recombinant human gp130 (Fig. 16B), indicating that the gp130-binding site of LIF was affected when LIF bound to h5D8.

[0174] Example 21 - LIF and LIFR expression in human tissues Quantitative real-time PCR was performed on many different types of human tissues to determine the expression levels of LIF and LIFR. The average expression levels shown in Figures 17A and 17B are expressed as copies per 100 ng of total RNA. Most tissues expressed at least 100 copies per 100 ng of total RNA. LIF mRNA expression was highest in human adipose tissue (mesenteric-ileal [1]), blood-vascular tissues (choroid-plexus [6] and mesentery [8]), and umbilical cord

[68] tissues, and lowest in brain tissues (cortex

[20] and substantia nigra

[28] ). LIFR mRNA expression was highest in human adipose tissue (mesenteric-ileal [1]), vascular tissues (lung [9]), brain tissue [11-28], and thyroid tissue

[66] , and lowest in PBMCs

[31] . LIF and LIFR mRNA expression levels in cynomolgus monkey tissues were similar to those observed in human tissues, where LIF expression was high in adipose tissue and LIFR expression was high in adipose tissue and low in PBMCs (data not shown).

[0175] The tissue numbering for Figures 17A and 17B is as follows: 1 - adipose (mesentery-ileum); 2 - adrenal gland; 3 - urinary bladder; 4 - urinary bladder (trigone); 5 - blood-vascular (cerebral: middle cerebral-artery); 6 - vascular (choroid-plexus); 7 - vascular (coronary artery); 8 - vascular (mesentery (colon)); 9 - vascular (lung); 10 - vascular (kidney); 11 - brain (amygdala); 12 - brain (caudate nucleus); 13 - brain (cerebellum); 14 Brain - (cortex: cingulate cortex - anterior); 15 - Brain (cortex: cingulate cortex - posterior); 16 - Brain (cortex: frontal cortex - lateral); 17 - Brain (cortex: frontal cortex - medial); 18 - Brain (cortex: occipital); 19 - Brain (cortex: parietal); 20 - Brain (cortex: temporal); 21 - Brain (dorsal raphe nucleus); 22 - Brain (hippocampus); 23 - Brain (hypothalamus: anterior); 24 - Brain (hypothalamus: posterior); 25 - Brain (locus coeruleus); 26 - Brain (medulla oblongata); 27 - Brain (nucleus accumbens); 28 - Brain (substantia nigra); 29 - breast; 30 - cecum; 31 - peripheral blood mononuclear cells (PBMCs); 32 - colon; 33 - dorsal root ganglion (DRG); 34 - duodenum; 35 - fallopian tube; 36 - gallbladder; 37 - heart (left atrium); 38 - heart (left ventricle); 39 - Ileum; 40 - Jejunum; 41 - Kidney (cortex); 42 - Kidney (medulla); 43 - Kidney (pelvis); 44 - Liver (parenchyma); 45 - Liver (main bronchi); 46 - Liver (tertiary bronchi); 47 - Lung (parenchyma); 48 - Lymph gland (tonsil); 49 - Muscle (skeletal muscle); 50 - Esophagus; 51 - Ovary; 52 - Pancreas; 53 - Pineal gland; 54 - Pituitary gland ;55-Placenta;56-Prostate;57-Rectum;58-Skin (foreskin);69-Spinal cord;60-Spleen (parenchyma);61-Stomach (antrum);62-Stomach (body);63-Stomach (fundus);64-Stomach (pyloric duct);65-Testes;66-Thyroid gland;67-Trachea;68-Umbilical cord;69-Ureter;70-Uterus (cervix);71-Uterus (muscle layer);and 72-Vas deferens.

[0176] Example 22 - h5D8 in combination with cisplatin, doxorubicin, or paclitaxel in a subcutaneous tumor xenograft model The efficacy of h5D6 in combination with cisplatin, doxorubicin, or paclitaxel was evaluated in a subcutaneously implanted CT26 model. CT26 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and penicillin / streptomycin (both 100 μg / ml). BALB / c mice were implanted subcutaneously at 10,000 cells per mouse, and treated with h5D8 (15 mg / kg) twice weekly and / or one of the following: cisplatin (10 mg / kg) once weekly, paclitaxel (15 mg / kg) twice weekly, or doxorubicin (4 mg / kg) once weekly. There were 12 mice per treatment cohort. Tumor volume and mouse weight were measured twice weekly using calipers.

[0177] As shown in Figure 18, mice treated with the combination of cisplatin and h5D8 exhibited reduced CT26 tumor growth compared to mice treated with cisplatin or h5D8 alone. As shown in Figure 19A, mice treated with the combination of doxorubicin and h5D8 did not exhibit reduced CT26 tumor growth compared to mice treated with doxorubicin or h5D8 alone. As shown in Figure 19B, mice treated with the combination of paclitaxel and h5D8 did not exhibit reduced CT26 tumor growth compared to mice treated with paclitaxel or h5D8 alone. Tumor volumes for all animals in all treatment groups measured at the 21-day endpoint are shown in Table 10.

[0178] [Table 10]

[0179] Example 23 - Combination of h5D8 and cisplatin in an intradermal tumor implant model The efficacy of h5D6 in combination with cisplatin, doxorubicin, or paclitaxel was evaluated in an intradermal CT26 model. CT26 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and penicillin / streptomycin (both at 100 μg / ml). BALB / c mice were intradermally implanted at 50,000 cells / mouse, and treated with h5D8 (15 mg / kg) twice weekly and / or cisplatin (10 mg / kg) once weekly. There were 12 mice per treatment cohort. Tumor volume and mouse weight were measured twice weekly using calipers.

[0180] As shown in Figure 20, mice treated with a combination of cisplatin and h5D8 exhibited reduced CT26 tumor growth when compared to mice treated with cisplatin or h5D8 alone. Tumor volumes for all animals in all treatment groups measured at the 21 day endpoint are shown in Table 11.

[0181] [Table 11]

[0182] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous changes, modifications, and substitutions will immediately occur to those skilled in the art that do not depart from the invention. It will be understood that various alternatives to the embodiments of the invention described herein can be employed in practicing the invention.

[0183] All publications, patent applications, issued patents, and / or documents mentioned herein are incorporated by reference herein to the same extent as if each individual publication, patent application, issued patent, and / or other document was specifically and individually indicated to be incorporated by reference in its entirety. Definitions contained in the text incorporated by reference are excluded to the extent they conflict with definitions in this disclosure.

[0184] array [Table 12]

[0185] [Table 13]

[0186] [Table 14]

[0187] [Table 15]

Claims

1. 1. A pharmaceutical composition for use in a method of treating an individual with cancer, said method comprising administering an effective amount of: a) a leukemia inhibitory factor (LIF) binding antibody, i) an immunoglobulin heavy chain complementarity determining region 1 (VH-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:2; ii) an immunoglobulin heavy chain complementarity determining region 2 (VH-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:5; iii) an immunoglobulin heavy chain complementarity determining region 3 (VH-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:6; iv) an immunoglobulin light chain complementarity determining region 1 (VL-CDR1) comprising the amino acid sequence set forth in SEQ ID NO: 10; v) an immunoglobulin light chain complementarity determining region 2 (VL-CDR2) comprising the amino acid sequence set forth in SEQ ID NO: 12; and vi) a LIF-binding antibody comprising an immunoglobulin light chain complementarity-determining region 3 (VL-CDR3) comprising the amino acid sequence set forth in SEQ ID NO: 13; and b) Platinum-based anticancer agents to an individual having said cancer, wherein said LIF-binding antibody is administered at a dose of 1500 mg, wherein said platinum-based anti-cancer agent is administered at a dose of 1 to 100 mg / m 2 , and wherein said pharmaceutical composition comprises said LIF-binding antibody and / or said platinum-based anti-cancer agent.

2. The pharmaceutical composition of claim 1, wherein the platinum-based anti-cancer agent is administered at a dose of 85 mg / m 2 .

3. The pharmaceutical composition of claim 1 , wherein the anti-LIF binding antibody comprises at least one framework region derived from a human antibody framework region.

4. The pharmaceutical composition of claim 1 , wherein the LIF-binding antibody is humanized.

5. The pharmaceutical composition of claim 1, wherein the LIF-binding antibody is deimmunized.

6. The pharmaceutical composition of claim 1, wherein the LIF-binding antibody comprises two immunoglobulin heavy chains and two immunoglobulin light chains.

7. The pharmaceutical composition of claim 1 , wherein the LIF-binding antibody is an IgG antibody.

8. The LIF-binding antibody is Fab, F(ab) ２ , a single domain antibody, a single chain variable fragment (scFv), or a nanobody.

9. the LIF-binding antibody a) an immunoglobulin heavy chain variable region (VH) sequence having an amino acid sequence at least 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 42; and b) an immunoglobulin light chain variable region (VL) sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO:46; 2. The pharmaceutical composition of claim 1, comprising:

10. 10. The pharmaceutical composition of claim 9, wherein the VH sequence is identical to the amino acid sequence set forth in SEQ ID NO: 42; and the VL sequence is identical to the amino acid sequence set forth in SEQ ID NO:

46.

11. the LIF-binding antibody a) an immunoglobulin heavy chain sequence having an amino acid sequence at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:58; and b) an immunoglobulin light chain sequence having an amino acid sequence that is at least about 80%, 90%, 95%, 97%, 98%, or 99% identical to the amino acid sequence set forth in SEQ ID NO:

62.

2. The pharmaceutical composition of claim 1, comprising:

12. The pharmaceutical composition of claim 1, wherein the LIF-binding antibody binds with a K D of less than about 200 picomolar.

13. The LIF-binding antibody has a K of less than about 100 picomolar. Ｄ The pharmaceutical composition of claim 1 , wherein the compound binds to the hydroxyl group.

14. 2. The pharmaceutical composition of claim 1, wherein the platinum-based anti-cancer agent comprises cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenatriplatin, picoplatin, satraplatin, or a combination thereof.

15. The pharmaceutical composition according to claim 14, wherein the platinum-based anti-cancer agent is cisplatin.

16. 2. The pharmaceutical composition of claim 1, wherein the cancer comprises advanced solid tumors, glioblastoma, gastric cancer, skin cancer, prostate cancer, pancreatic cancer, breast cancer, testicular cancer, thyroid cancer, head and neck cancer, liver cancer, kidney cancer, esophageal cancer, ovarian cancer, colon cancer, lung cancer, lymphoma, or soft tissue cancer.

17. 17. The pharmaceutical composition of claim 16, wherein the cancer comprises non-small cell lung cancer, ovarian epithelial cancer, or pancreatic adenocarcinoma.

18. 10. The pharmaceutical composition of claim 1, wherein the cancer is resistant to treatment with a therapeutic amount of an inhibitor of an LIF-binding antibody.

19. 2. The pharmaceutical composition of claim 1, wherein the cancer is resistant to treatment with a therapeutic amount of a platinum-based anti-cancer agent.

20. The pharmaceutical composition of claim 1, wherein the leukemia inhibitory factor (LIF)-binding antibody and the platinum-based anti-cancer agent are administered separately.

21. The pharmaceutical composition of claim 1 , wherein the LIF-binding antibody and the platinum-based anti-neoplastic agent are administered on different schedules.

22. The pharmaceutical composition of claim 1 , wherein the LIF-binding antibody and the platinum-based anti-neoplastic agent are administered in a single composition.

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