Anti-IL-11Rα Antibody

Abstract

Provided are antibodies and antigen-binding fragments thereof that bind to human interleukin-11 receptor subunit alpha (IL-11Rα) and related compositions, which may be used in any of a variety of therapeutic or diagnostic methods, including cancer, inflammatory diseases, autoimmune diseases, and other treatments or diagnoses.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 238,443, filed August 30, 2021; and U.S. Provisional Patent Application No. 63 / 334,923, filed April 26, 2022, each of which is incorporated by reference in its entirety.

[0002] Description of sequence listing The sequence listing XML associated with this application is provided in XML file format and is incorporated herein by reference. The name of the XML file containing the sequence listing XML is LASS_005_02WO_ST26.xml. The XML file is approximately 328,887 bytes, was created on August 15, 2022, and has been submitted electronically via the USPTO Patent Center.

[0003] The present disclosure relates to antibodies and antigen-binding fragments thereof that bind to human interleukin-11 receptor subunit alpha (IL-11Rα) and related compositions, which may be used in any of a variety of therapeutic or diagnostic methods, including cancer, inflammatory diseases, autoimmune diseases, and other treatments or diagnoses. [Background technology]

[0004] 2. Description of Related Art Interleukin-11 (IL-11) is a member of the IL-6 family and plays a prominent role in chronic inflammation, autoimmunity, cancer, and other diseases. It also plays a critical role in the initiation and maintenance of chronic fibrotic responses. Thus, IL-11 signaling inhibitors represent a promising therapeutic approach for treating a variety of diseases, including inflammatory diseases, autoimmune diseases, chronic fibrotic diseases, and cancer.

[0005] Exemplary IL-11 signaling inhibitors under development include antibodies that bind to interleukin-11 receptor subunit alpha (IL-11Rα) (see, e.g., U.S. Patent Nos. 9,796,782; and 9,340,618). However, there is a need in the art for anti-IL-11Rα antibodies with increased potency and optimal developability characteristics. Summary of the Invention

[0006]

[0013] Embodiments of the present disclosure include isolated antibodies, or antigen-binding fragments thereof, that bind to interleukin-11 receptor subunit alpha (IL-11Rα), wherein at least one antibody, or antigen-binding fragment thereof, comprises: Complementarity determining region V selected from Table A1 H CDR1, V H CDR2, and V H A heavy chain variable region (V) containing a CDR3 sequence and a variant thereof that specifically binds to IL-11Rα. H ); and Complementarity determining region V selected from Table A1 L CDR1, V L CDR2, and V L A light chain variable region (V) containing a CDR3 sequence and a variant thereof that specifically binds to IL-11Rα. L ).

[0007] In some embodiments: V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 1 to 3, respectively, and V L CDR1, V L CDR2, and V L the CDR3 sequences include SEQ ID NOs: 4 to 6, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 7 to 9, respectively, and V L CDR1, V L CDR2, and V LCDR3 sequences include SEQ ID NOs: 10 to 12, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 13 to 15, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 16 to 18, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 19 to 21, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 22 to 24, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 25 to 27, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 28 to 30, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 31 to 33, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 34 to 36, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 34 to 39, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 40 to 42, respectively; V H CDR1, V H CDR2, and V HThe CDR3 sequences include SEQ ID NOs: 43 to 45, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 46 to 48, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 49 to 51, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 52 to 54, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 55 to 57, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 58 to 60, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 61 to 63, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 64 to 66, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 67 to 69, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 70 to 72, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 73 to 75, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 76 to 78, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 79 to 81, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 82 to 84, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 85 to 87, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 88 to 90, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 91 to 93, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 94 to 96, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 97 to 99, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 100 to 102, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 103 to 105, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 106 to 108, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 109 to 111, respectively, and V LCDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 112-114, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 115 to 117, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 118-120, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 121 to 123, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 124 to 126, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 127 to 129, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 130 to 132, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 133 to 135, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 136 to 138, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 139 to 141, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 142 to 144, respectively; V HCDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 145 to 147, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 148 to 150, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 151 to 153, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 154 to 156, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 157 to 159, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 160 to 162, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 163 to 165, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 166-168, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 169 to 171, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 172 to 174, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 175 to 177, respectively, and V L CDR1, VL CDR2, and V L CDR3 sequences include SEQ ID NOs: 178 to 180, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 181 to 183, respectively, and V L CDR1, V L CDR2, and V L the CDR3 sequences comprise SEQ ID NOs: 184 to 186, respectively; or V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 187 to 189, respectively, and V L CDR1, V L CDR2, and V L The CDR3 sequences include SEQ ID NOs: 190 to 192, respectively.

[0008] In some embodiments, V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally H In some embodiments, V has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions. L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, and optionally L has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in the framework regions.

[0009] In certain embodiments: V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 193, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 194; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 195, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 196; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 197, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 198; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 199, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 200; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 201, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:202; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 203, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:204; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 205, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:206; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 207, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:208; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 209, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:210; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 211, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:212; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 213, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:214; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 215, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:216; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 217, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:218; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 219, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 220; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 221, and V Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:222; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 223, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:224; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 225, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:226; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 227, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:228; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 229, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:230; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 231, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:232; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 233, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:234; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 235, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:236; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 237, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:238; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 239, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:240; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 241, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:242; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 243, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:244; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 245, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:246; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 247, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:248; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 249, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 250; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 251, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:252; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 253, and L comprises a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 254; or V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 255, and L comprises a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:256.

[0010] In some embodiments, the isolated antibody, or antigen-binding fragment thereof, binds to human IL-11Rα (see Table B1). In certain embodiments, the isolated antibody, or antigen-binding fragment thereof, binds to fibronectin domain III of human IL-11Rα, or to approximately residues 112-219 of SEQ ID NO: 257. In some embodiments, the isolated antibody, or antigen-binding fragment thereof, has one or more of the following characteristics: have a binding affinity for human IL-11Rα of less than about 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 40, or 30 pM, and optionally have an increased binding affinity for human IL-11Rα compared to that of the TS7 and 8E2 antibodies; antagonize the binding and / or signaling activity between IL-11Rα and IL-11, and optionally have increased potency as an IL-11 signaling antagonist compared to that of the TS7 and 8E2 antibodies; optionally reduce IL-11Rα / gp130 dimerization or complex formation in a cell-based assay; and / or optionally have a reduced V-terminal binding affinity compared to that of the TS7 and 8E2 antibodies. L It has reduced N-linked glycosylation in CDR3.

[0011] In some embodiments, the isolated antibody, or antigen-binding fragment thereof, comprises an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or hybrids and / or variants thereof. In some embodiments, the isolated antibody, or antigen-binding fragment thereof, comprises an IgG Fc domain with high effector function in humans, optionally an IgG1 or IgG3 Fc domain. In some embodiments, the isolated antibody, or antigen-binding fragment thereof, comprises an IgG Fc domain with low effector function in humans, optionally an IgG2 or IgG4 Fc domain. In certain embodiments, the isolated antibody, or antigen-binding fragment thereof, optionally comprises a human IgG1 or IgG4 Fc domain selected from Table F1.

[0012] In some embodiments, the isolated antibody, or antigen-binding fragment thereof, is a monoclonal antibody. In some embodiments, the isolated antibody, or antigen-binding fragment thereof, comprises a humanized antibody, wherein the antibody, or antigen-binding fragment thereof, is a humanized monoclonal antibody comprising a human IgG4 Fc domain with a S228P mutation (EU numbering). In some embodiments, the isolated antibody, or antigen-binding fragment thereof, is selected from an Fv fragment, a single-chain Fv (scFv) polypeptide, an adnectin, anticalin, an aptamer, an avimer, a camelid antibody, a designed ankyrin repeat protein (DARPin), a minibody, a nanobody, and a unibody.

[0013] Also included are isolated polynucleotides encoding the isolated antibodies, or antigen-binding fragments thereof, described herein, expression vectors comprising the isolated polynucleotides, and isolated host cells comprising the vectors (S).

[0014] Certain embodiments relate to pharmaceutical compositions comprising an isolated antibody, or antigen-binding fragment thereof, described herein, and a pharma- ceutical acceptable carrier. In some embodiments, the composition has a purity of at least about 80%, 85%, 90%, 95%, 98%, or 99% on a protein basis with respect to at least one antibody or antigen-binding fragment, and is substantially free of aggregates and endotoxins. In some embodiments, the composition optionally comprises a V L and has reduced or undetectable heterogeneity (optionally compared to the TS7 and 8E2 antibodies) of N-linked glycosylation in the CDR3 sequences. In some embodiments, the composition is a sterile, injectable solution, optionally suitable for intravenous, intramuscular, subcutaneous, or intraperitoneal administration.

[0015] Also included are methods of treating a disease or condition in a subject in need thereof comprising administering to the subject a pharmaceutical composition described herein. In some embodiments, the disease or condition is an IL-11-associated or IL-11-mediated disease or condition. In some embodiments, the disease or condition is cancer, an inflammatory disease, an autoimmune disease, a wasting disease, a bone disease, or a fibrosis.

[0016] In some embodiments, the disease is cancer, optionally a cancer that expresses or overexpresses IL-11Rα and / or IL-11, optionally in which the cancer exhibits IL-11Rα / IL-11 dependent growth, adhesion, migration, invasion, and / or chemotherapy resistance. In some embodiments, the cancer is bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma), liver cancer, or tumor. In some embodiments, the cancer is selected from one or more of: myeloid leukemia (hepatocellular carcinoma), sarcoma, B-cell malignancies, breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, uterine cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer. In some embodiments, the cancer is a metastatic cancer, optionally a metastatic cancer that has metastasized to bone.

[0017] In certain embodiments, the inflammatory disease is selected from one or more of inflammation of the airways or lungs (e.g., inflammatory lung disease), asthma, rhinitis, chronic obstructive pulmonary disorder (COPD), dermatitis, psoriasis, hepatitis, gastritis, irritable bowel syndrome (IBS), ulcerative colitis, Crohn's disease, colitis, diverticulitis, lupus erythematosus, nephritis, Parkinson's disease, multiple sclerosis (MS), Alzheimer's disease, arthritis, rheumatoid arthritis, sepsis, infection-induced inflammation, cardiovascular disease, such as atherosclerosis and vasculitis, diabetes, and gout.

[0018] In certain embodiments, the autoimmune disease is selected from the group consisting of arthritis (including rheumatoid arthritis, reactive arthritis), systemic lupus erythematosus (SLE), psoriasis, inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, encephalomyelitis, uveitis, myasthenia gravis, multiple sclerosis, insulin-dependent diabetes mellitus, Addison's disease, celiac disease, chronic fatigue syndrome, autoimmune hepatitis, autoimmune alopecia, ankylosing spondylitis, fibromyalgia, pemphigus vulgaris, Sjogren's syndrome, Kawasaki disease, hyperthyroidism / Graves' disease, hypothyroidism / Hashimoto's disease, endometriosis, scleroderma, pernicious anemia, Goodpa The present invention is directed to a method for treating or preventing an autoimmune disease, comprising administering to a patient a suitable therapeutic agent, such as a pulmonary artery disease, angioedema, pulmonary edema, or a combination of these conditions, selected from one or more of: Schürer's syndrome, Guillain-Barré syndrome, Wegener's disease, glomerulonephritis, aplastic anemia (including patients with multiply transfused aplastic anemia), paroxysmal nocturnal hemoglobinuria, myelodysplastic syndrome, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, Evan's syndrome, factor VIII inhibition syndrome, systemic vasculitis, dermatomyositis, polymyositis, rheumatic fever, autoimmune lymphoproliferative syndrome (ALPS), autoimmune bullous pemphigoid, Parkinson's disease, sarcoidosis, vitiligo, primary biliary cirrhosis, and autoimmune myocarditis.

[0019] In some embodiments, the wasting disease is selected from one or more of cachexia, optionally cachexia associated with cancer or renal failure, and sarcopenia.

[0020] In some embodiments, the bone disease is selected from osteoporosis (including postmenopausal osteoporosis), bone fractures, Paget's disease of bone, and bone resorption / damage associated with cancer or cancer treatments, including chemotherapy, hormone ablation, and hormone blockade.

[0021] In some embodiments, the fibrosis is selected from pulmonary, cardiovascular, liver, brain, joint (optionally knee, hip, ankle, ankle, shoulder, elbow, wrist, hand, or spine), intestine, skin, kidney, liver, thyroid, bone marrow, retroperitoneum, and eye fibrosis. In certain embodiments, the pulmonary fibrosis is selected from fibrothorax, pulmonary fibrosis (optionally cystic fibrosis or interstitial lung disease (ILD)), autosomal recessive disorders, optionally Hermansky-Pudlak syndrome, and radiation-induced lung injury. In certain embodiments, the ILD is an idiopathic ILD, optionally selected from idiopathic pulmonary fibrosis (IPF), desquamative interstitial pneumonia (DIP), acute interstitial pneumonia (AIP) or Hamman-Rich syndrome, nonspecific interstitial pneumonia (NSIP), respiratory bronchiolitis-associated interstitial lung disease (RB-ILD), cryptopathic organizing pneumonia (COP), and lymphocytic interstitial pneumonia (LIP). In certain embodiments, the ILD is a secondary ILD, optionally selected from ILDs associated with connective tissue and autoimmune diseases (optionally sarcoidosis, rheumatoid arthritis, systemic lupus erythematosus, systemic sclerosis, polymyositis, dermatomyositis, or antisynthetase syndrome), inhalants (optionally silicosis, asbestosis, beryllium disease, industrial printing chemicals, or chronic hypersensitivity pneumonitis), drugs (optionally antibiotics, chemotherapeutic agents, or antiarrhythmic agents), infections (optionally SARS CoV-2, atypical pneumonia, Pneumocystis pneumonia, tuberculosis, Chlamydia trachomatis, or respiratory syncytial virus), malignancies (optionally lymphangitis carcinomatosis), and pediatric ILDs (optionally developmental disorders, growth abnormalities, alveolar hypoplasia, infantile conditions of unknown etiology, and alveolar surfactant area-associated ILD).

[0022] In some embodiments, the fibrosis of the cardiovascular system is myocardial fibrosis (eg, interstitial fibrosis or replacement fibrosis). [Brief description of the drawings]

[0023] [Figure 1A] 1A-1B show the gross appearance of lung tissue from donor 1 (1A) and donor 2 (1B). [Figure 1B] Same as above. [Figure 2A]Figures 2A-2B show hematoxylin and eosin staining of fibrotic lung tissue from donor 1 (2A, upper subpleural; bottom center) and donor 2 (2B, upper subpleural; bottom center). Staining was performed on tissue adjacent to the PCLS tissue core. Representative overview photographs are shown. [Figure 2B] Same as above. [Figure 3A] Figures 3A-3B show tissue viability of untreated (medium) and LPS-treated precision-cut lung slices (PCLS) from donor 1 (3A) and donor 2 (3B). LDH release is given as % of Triton lysis control (set to 100%). Bars indicate mean ± SD and dots represent technical replicates (duplicate wells with two PCLS each). For donor 2 (3B), supernatants of duplicate wells were pooled before measurement. Region 1 is subpleural and region 2 is central. [Figure 3B] Same as above. [Figure 4A] Figures 4A-4B show IL-1β upregulation in LPS-treated PCLS from donor 1 (4A) and donor 2 (4B). IL-1β was determined in PCLS lysates by ELISA and normalized to total protein concentration of the tissue. Data are shown as mean ± SD. For each condition, lysates from duplicate wells (each with two PCLS) were pooled before measurement. Region 1 is subpleural and region 2 is central. [Figure 4B] Same as above. [Diagram 5] Figure 5 shows collagen VI formation assessed by PRO-C6 across all donors and regions. Region 1 is subpleural and region 2 is central. Data are presented as a percentage of no treatment (medium) control and shown as individual data points for three replicates from each region from each donor (12 in total), with indication of median and interquartile range. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] The present disclosure relates to antibodies, and antigen-binding fragments thereof, that specifically bind to interleukin-11 receptor subunit alpha (IL-11Rα), in particular antibodies with epitope specificity and improved characteristics. Examples of such improved characteristics include increased binding affinity for IL-11Rα, increased potency as interleukin-11 (IL-11) signaling inhibitors / antagonists, and improved developability, such as increased manufacturing uniformity due to reduced heterogeneity at potential N-linked glycosylation sites. Some embodiments thus include certain humanized antibodies and fragments thereof that are capable of binding to IL-11Rα, blocking IL-11Rα binding to its ligand IL-11, and inhibiting downstream cell signaling and biological effects. In certain embodiments, the anti-IL-11Rα antibodies, or antigen-binding fragments thereof, are IL-11Rα antagonists or inhibitors.

[0025] The IL-11Rα antagonist antibodies described herein are useful in the treatment and prevention of a variety of diseases and conditions, including those associated with or mediated by IL-11 signaling, such as cancer, autoimmune diseases, and inflammatory diseases. Some embodiments thus relate to the use of anti-IL-11Rα antibodies, or antigen-binding fragments thereof, for the diagnosis, evaluation, and treatment of diseases and conditions, including those associated with IL-11 activity or its aberrant expression.

[0026] The practice of the present disclosure will employ, unless specifically indicated to the contrary, conventional methods of virology, immunology, microbiology, molecular biology, and recombinant DNA technology within the skill of the art, many of which are described below for illustrative purposes. Such techniques are explained fully in the literature. See, for example, Current Protocols in Molecular Biology or Current Protocols in Immunology, John Wiley & Sons, New York, NY (2009); Ausubel et al., Short Protocols in Molecular Biology, 3 rded., Wiley & Sons, 1995; Sambrook and Russell, Molecular Cloning: A Laboratory Manual (3rd Edition, 2001); Maniatis et al. Molecular Cloning: A Laboratory Manual (1982); DNA Cloning: A Practical Approach, vols. I & II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait, ed., 1984); Nucleic Acid Hybridization (B. Hames & S. Higgins, eds., 1985); Transcription and Translation (B. Hames & S. Higgins, eds., 1984); Animal Cell Culture (R. Freshney, ed., 1986); Perbal, A Practical Guide to Molecular Cloning (1984), and other similar references.

[0027] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the content clearly dictates otherwise.

[0028] By "about" is meant an amount, level, value, number, frequency, percentage, dimension, size, amount, weight, or length that varies by 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1% from the reference amount, level, value, number, frequency, percentage, dimension, size, amount, weight, or length.

[0029] The term "antigen" refers to a molecule or a portion of a molecule that can be bound by a selective binding agent, such as an antibody, and can be used in an animal to generate an antibody capable of binding to an epitope of that antigen. An antigen may have one or more epitopes. As used herein, the term "antigen" includes a substance that, under suitable conditions, is capable of inducing an immune response against the substance and reacting with the products of the immune response. For example, an antigen can be recognized by an antibody (humoral immune response) or a sensitized T lymphocyte (T helper or cell-mediated immune response), or both. Antigens can be soluble substances, such as toxins and foreign proteins, or microparticles, such as bacteria and tissue cells; however, only parts of a protein or polysaccharide molecule, known as an antigenic determinant (epitope), combine with an antibody or a specific receptor on a lymphocyte. More broadly, the term "antigen" includes any substance to which an antibody binds or for which an antibody is desired, regardless of whether the substance is immunogenic or not. For such antigens, antibodies can be identified by recombinant methods, independent of any immune response.

[0030] "Antagonist" refers to an agent (e.g., an antibody) that interferes with or otherwise reduces the physiological action of another agent or molecule. In some instances, antagonists specifically bind to other agents or molecules. Included are full and partial antagonists.

[0031] "Agonist" refers to an agent (e.g., an antibody) that increases or enhances the physiological effect of another agent or molecule. In some instances, agonists specifically bind to other agents or molecules. Included are full and partial agonists.

[0032] As used herein, the term "amino acid" is intended to mean both natural and unnatural amino acids, as well as amino acid analogs and mimetics. Natural amino acids include the 20 (L) amino acids utilized during protein biosynthesis, as well as others, such as 4-hydroxyproline, hydroxylysine, desmosine, isodesmosine, homocysteine, citrulline, and ornithine. Unnatural amino acids include, for example, the (D)-amino acids, norleucine, norvaline, p-fluorophenylalanine, ethionine, and the like, which are known to those of skill in the art. Amino acid analogs include modified forms of natural and unnatural amino acids. Such modifications can include, for example, the substitution or replacement of chemical groups and moieties on the amino acid, or can be by derivatization of the amino acid. Amino acid mimetics include, for example, organic structures that exhibit functionally similar properties, such as the charge and charge spacing characteristics of the reference amino acid. For example, an organic structure that mimics arginine (Arg or R) will have a positively charged moiety positioned in a similar molecular space and with the same degree of flexibility as the e-amino group of the side chain of the natural Arg amino acid. Mimetics also include structures that are constrained to maintain optimal spacing and charge interactions of the amino acids or amino acid functional groups. Those skilled in the art will know or be able to determine which structures constitute functionally equivalent amino acid analogs and amino acid mimetics.

[0033] As used herein, the term "antibody" includes not only intact polyclonal or monoclonal antibodies, but also fragments thereof (e.g., dAb, Fab, Fab', F(ab')2, Fv, etc.), single chain (scFv), synthetic variants thereof, naturally occurring variants, fusion proteins comprising an antibody portion with an antigen-binding fragment of the required specificity, humanized antibodies, chimeric antibodies, and any other modified configurations of immunoglobulin molecules that contain an antigen-binding site or fragment (epitope recognition site) of the required specificity. Specific features and characteristics of antibodies (and antigen-binding fragments thereof) are described in more detail herein.

[0034] The antibody or antigen-binding fragment can be of essentially any type. As is well known in the art, an antibody is an immunoglobulin molecule capable of specific binding to a target, such as an immune checkpoint molecule, through at least one epitope recognition site located in the variable region of the immunoglobulin molecule.

[0035] As used herein, the term "antigen-binding fragment" refers to a polypeptide fragment that contains at least one CDR of an immunoglobulin heavy and / or light chain that binds to an antigen of interest. In this regard, an antigen-binding fragment of an antibody described herein is a V-type polypeptide fragment from an antibody that binds to a target molecule. H and V L It may comprise one, two, three, four, five or all six CDRs of the sequence.

[0036] The binding properties of antibodies and antigen-binding fragments thereof can be quantified using methods well known in the art (see Davies et al., Annual Rev. Biochem. 59:439-473, 1990). In some embodiments, an antibody or antigen-binding fragment thereof binds a target molecule, e.g., an IL-11Rα polypeptide or an epitope or complex thereof, with a binding affinity of about ≦10 -7 M~about 10 -8 M. In some embodiments, the equilibrium dissociation constant is about ≦10 -9 M~approx.≦10 -10 In certain exemplary embodiments, the antibody or antigen-binding fragment thereof has an affinity (K) for a target molecule (to which it specifically binds) of less than about, at least about, or about 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, or 50 nM. D or EC 50 ).

[0037] A molecule, such as a polypeptide or an antibody, is said to exhibit "specific binding" or "preferential binding" if it reacts or associates with a particular cell, substance, or particular epitope more frequently, more rapidly, with greater duration, and / or with greater affinity than it does with alternative cells or substances or epitopes. An antibody "specifically binds" or "preferentially binds" to a target molecule or epitope if it binds, for example, by a statistically significant amount, with greater affinity, avidity, more readily, and / or with longer duration than it binds to other substances or epitopes. Typically, one member of a pair of molecules exhibiting specific binding has an area, or cavity, on its surface that specifically binds to, and is therefore complementary to, a particular spatial and / or polar structure of the other member of the pair of molecules. In this way, the members of the pair have the property of specifically binding to each other. For example, an antibody that specifically or preferentially binds to a specific epitope is one that binds to that specific epitope with greater affinity, avidity, more readily, and / or with longer duration than it binds to other epitopes. It is also understood by reading this definition that, for example, an antibody (or moiety or epitope) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. The term is also applicable, for example, when an antibody is specific for a particular epitope carried by multiple antigens, in which case a specific binding member carrying an antigen-binding fragment or domain can bind to a variety of antigens carrying the epitope; for example, it may be cross-reactive to multiple different forms of a target antigen from multiple species that share a common epitope.

[0038] Immunological binding generally refers to the type of non-covalent interaction that occurs between an immunoglobulin molecule and an antigen for which the immunoglobulin is specific, for example, by way of illustration and not limitation, as a result of electrostatic, ionic, hydrophilic and / or hydrophobic attractions or repulsions, steric forces, hydrogen bonding, van der Waals forces, and other interactions. The strength, or affinity, of an immunological binding interaction is determined by the dissociation constant (K D) and smaller K D represents a greater affinity. The immunological binding properties of a selected polypeptide can be quantified using methods well known in the art. One such method involves measuring the rates of antigen-binding site / antigen complex formation and dissociation, where those rates depend on the concentrations of the complex partners, the affinity of the interaction, and geometric parameters that affect the rates equally in both directions. In this way, both the "on-rate constant" (Kon) and the "off-rate constant" (Koff) can be determined by calculation of the concentrations and the actual rates of association and dissociation. The ratio of Koff / Kon allows the release of all parameters that are not related to affinity, and thus the dissociation constant K D As used herein, the term "affinity" includes the equilibrium constant for the reversible binding of two agents, K D or EC 50 The affinity of a binding protein to a ligand, such as the affinity of an antibody for an epitope, can be, for example, from about 100 nanomolar (nM) to about 0.1 nM, from about 100 nM to about 1 picomolar (pM), or from about 100 nM to about 1 femtomolar (fM). As used herein, the term "avidity" refers to the resistance of a complex of two or more agents to dissociation upon dilution. In some embodiments, affinity is measured using the half maximal effective concentration (EC 50 EC is expressed in terms of the concentration of an agent, such as an antibody or anti-IL-11Rα antibody, as disclosed herein, that induces a response halfway between the baseline and maximum after a specified exposure time. 50 is commonly used as a measure of antibody potency.

[0039] Antibodies can be prepared by any of a variety of techniques known to those of skill in the art. See, e.g., Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988. Monoclonal antibodies specific for a polypeptide of interest can be prepared, for example, using the techniques of Kohler and Milstein, Eur. J. Immunol. 6:511-519, 1976, and improvements thereon. Also included are methods of expressing human antibodies using transgenic animals, such as mice. See, e.g., Neuberger et al., Nature Biotechnology 14:826, 1996; Lonberg et al., Handbook of Experimental Pharmacology 113:49-101, 1994; and Lonberg et al., Internal Review of Immunology 13:65-93, 1995. A specific example includes the VELOCIMMUNE® platform by REGENEREX® (see, eg, US Pat. No. 6,596,541).

[0040] Antibodies can also be generated or identified by using phage display libraries or yeast display libraries (see, e.g., U.S. Pat. No. 7,244,592; Chao et al., Nature Protocols. 1:755-768, 2006). Non-limiting examples of available libraries include cloned or synthetic libraries, such as the Human Combinatorial Antibody Library (HuCAL), in which the structural diversity of the human antibody repertoire is represented by seven heavy and seven light chain variable region genes. The combination of these genes results in 49 frameworks in the master library. By stacking highly variable gene cassettes (CDRs = complementarity determining regions) on these frameworks, the vast human antibody repertoire can be reproduced. Also included are human libraries designed using human donor-supplied fragments encoding the light chain variable region, synthetic DNA encoding the diversity of the heavy chain CDR-3, heavy chain CDR-1, and synthetic DNA encoding the diversity of the heavy chain CDR-2. Other libraries suitable for use will be apparent to those skilled in the art.

[0041] In certain embodiments, the antibodies and antigen-binding fragments thereof described herein comprise a set of heavy and light chain CDRs, each interposed between a set of heavy and light chain framework regions (FRs), which provide support for the CDRs and define the spatial relationship of the CDRs to each other. As used herein, the term "CDR set" refers to the three hypervariable regions of a heavy or light chain V region. Proceeding from the N-terminus of the heavy or light chain, these regions are denoted as "CDR1", "CDR2", and "CDR3", respectively. An antigen-binding site thus comprises six CDRs, including a set of CDRs from each of the heavy and light chain V regions. A polypeptide comprising a single CDR (e.g., CDR1, CDR2, or CDR3) is referred to herein as a "molecular recognition unit". Crystallographic analysis of numerous antigen-antibody complexes has demonstrated that the amino acid residues of the CDRs form extensive contacts with bound antigen, with the most extensive antigen contacts being with the heavy chain CDR3. Thus, the molecular recognition units are primarily responsible for the specificity of the antigen-binding site.

[0042] As used herein, the term "FR set" refers to four adjacent amino acid sequences that frame the CDRs of a CDR set of a heavy or light chain V region. Some FR residues may contact the bound antigen; however, FRs are primarily involved in folding the V region into an antigen binding site, particularly the FR residues directly adjacent to the CDRs. Within the FRs, certain amino acid residues and certain structural features are highly conserved. In this regard, most V region sequences contain an internal disulfide loop of about 90 amino acid residues. When the V region folds into the binding site, the CDRs are presented as protruding loop motifs that form an antigen binding surface. Regardless of the exact CDR amino acid sequence, it is generally recognized that there are conserved structural regions of FRs in certain "canonical" structures that affect the folded shape of the CDR loops. In addition, certain FR residues are known to be involved in non-covalent interdomain contacts that stabilize the interaction of antibody heavy and light chains.

[0043] The structure and location of immunoglobulin variable domains can be determined by reference to Kabat, EA et al., Sequences of Proteins of Immunological Interest. 4th Edition. U.S. Department of Health and Human Services, 1987, and updates thereof.

[0044] Also included are monoclonal antibodies, which refer to homogeneous antibody populations composed of amino acids (natural and non-natural) involved in selective binding of an epitope. Monoclonal antibodies are highly specific and directed against a single epitope. The term "monoclonal antibody" encompasses intact and full-length monoclonal antibodies, but also fragments thereof (e.g., Fab, Fab', F(ab')2, Fv, etc.), single chain (scFv), variants thereof, fusion proteins containing antigen-binding portions, humanized monoclonal antibodies, chimeric monoclonal antibodies, and any other modified configuration of an immunoglobulin molecule containing an antigen-binding fragment (epitope recognition site) of the required specificity and ability to bind to the epitope. It is not intended to be limited with regard to the source of the antibody or the manner in which it is made (e.g., by hybridoma, phage selection, recombinant expression, transgenic animals). The term includes whole immunoglobulins, as well as the fragments and the like described above under the definition of "antibody."

[0045] The proteolytic enzyme papain preferentially cleaves IgG molecules to produce several fragments, two of which (F(ab) fragments) each contain a covalently linked heterodimer with an intact antigen binding site. The enzyme pepsin can cleave IgG molecules to provide several fragments, including the F(ab')2 fragment, which contains both antigen binding sites. Fv fragments for use according to certain embodiments can be produced by preferential proteolytic cleavage of IgM and, in rare cases, IgG or IgA immunoglobulin molecules. Fv fragments are, however, more commonly derived using recombinant techniques known in the art. Fv fragments comprise a non-covalent VH::VL heterodimer comprising an antigen-binding site that retains much of the antigen recognition and binding capabilities of a native antibody molecule (Inbar et al., PNAS USA. 69:2659-2662, 1972; Hochman et al., Biochem. 15:2706-2710, 1976; and Ehrlich et al., Biochem. 19:4091-4096, 1980). In some embodiments, the Fv is stabilized by other means, such as the incorporation of at least one disulfide bond (Worn & Pluckthun, J. Mol. Biol. 305, 989-1010, 2001).

[0046] In certain embodiments, single chain Fv (scFV) antibodies are contemplated. For example, kappabodies (Ill et al., Prot. Eng. 10:949-57, 1997); minibodies (Martin et al., EMBO J 13:5305-9, 1994); diabodies (Holliger et al., PNAS 90:6444-8, 1993); or Janusins ​​(Traunecker et al., EMBO J 10:3655-59, 1991; and Traunecker et al., Int. J. Cancer Suppl. 7:51-52, 1992) can be prepared using standard molecular biology techniques following the teachings of the present application for selecting antibodies with the desired specificity.

[0047] Single-chain Fv (scFv) polypeptides are covalently linked VH::VL heterodimers, which are expressed from gene fusions containing VH and VL coding genes linked by a peptide-encoding linker. Huston et al. (PNAS USA.85(16):5879-5883,1988). Numerous methods have been described for identifying chemical structures for converting the naturally aggregated, but chemically separated, light and heavy polypeptide chains from antibody V regions into scFv molecules that fold into a three-dimensional structure substantially similar to that of the antigen-binding site. See, for example, U.S. Patent Nos. 5,091,513 and 5,132,405 to Huston et al.; and U.S. Patent No. 4,946,778 to Ladner et al.

[0048] In certain embodiments, the antibodies or antigen-binding fragments described herein are in the form of "diabodies". Diabodies are multimers of polypeptides, each of which comprises a first domain comprising a binding region of an immunoglobulin light chain and a second domain comprising a binding region of an immunoglobulin heavy chain, the two domains being linked (e.g., by a peptide linker) but unable to associate with each other to form an antigen-binding site: the antigen-binding site is formed by association of a first domain of one polypeptide in the multimer with a second domain of another polypeptide in the multimer (WO94 / 13804). dAb fragments of antibodies consist of a VH domain (Ward et al., Nature 341:544-546, 1989). Diabodies and other multivalent or multispecific fragments can be constructed, for example, by gene fusion (see WO94 / 13804; and Holliger et al., PNAS USA. 90:6444-6448, 1993).

[0049] Also included are minibodies that contain an scFv linked to a CH3 domain (see Hu et al., Cancer Res. 56:3055-3061, 1996). See also Ward et al., Nature. 341:544-546, 1989; Bird et al., Science. 242:423-426, 1988; Huston et al., PNAS USA. 85:5879-5883, 1988); PCT / US92 / 09965; WO94 / 13804; and Reiter et al., Nature Biotech. 14:1239-1245, 1996.

[0050] Where bispecific antibodies are used, these may be conventional bispecific antibodies, which may be produced in a variety of ways (Holliger and Winter, Current Opinion Biotechnol. 4:446-449, 1993), for example prepared chemically or from hybridomas, or may be any of the bispecific antibody fragments mentioned above.

[0051] Bispecific diabodies, as opposed to bispecific whole antibodies, can also be particularly useful because they can be easily constructed and expressed in E. coli. Diabodies (and many other polypeptides, such as antibody fragments) of suitable binding specificity can be easily selected using phage display (WO94 / 13804) from libraries. If one arm of the diabody is kept constant, for example with specificity directed against antigen X, then the other arm can be varied to create a library from which antibodies of suitable specificity are selected. Bispecific whole antibodies can be generated by a number of methods, including knob-into-hole engineering (Brinkman & Kontermann, mAbs 9:182-212, 2017) (Ridgeway et al., Protein Eng. 9:616-621, 1996).

[0052] In certain embodiments, the antibodies or antigen-binding fragments described herein are in the form of UniBody®. UniBody® is an IgG4 antibody with the hinge region removed (see GenMab Utrecht, The Netherlands; see also US20090226421). This antibody technology creates a stable, smaller antibody format with a predicted longer therapeutic window than current small antibody formats. IgG4 antibodies are considered inert and thus do not interact with the immune system. Fully human IgG4 antibodies can be engineered by removing the hinge region of the antibody to obtain half-molecule fragments with different stability characteristics compared to the corresponding intact IgG4 (GenMab, Utrecht). By halving the IgG4 molecule, only one region remains on the UniBody® that can bind to the cognate antigen (e.g., disease target), and thus the UniBody® binds monovalently to only one site on the target cell. For certain cancer cell surface antigens, this monovalent binding will not stimulate cancer cells to grow as may be seen using bivalent antibodies with the same antigen specificity, and therefore UniBody® technology may offer a treatment option for some types of cancer that may be refractory to treatment with traditional antibodies. The small size of UniBody® may be of great benefit in treating some forms of cancer, but also allows for better distribution of the molecule to larger solid tumors, potentially increasing efficacy.

[0053] In certain embodiments, the antibodies and antigen-binding fragments described herein are in the form of nanobodies. Nanobodies are encoded by a single gene and are efficiently produced in almost all prokaryotic and eukaryotic hosts, such as E. coli (see U.S. Pat. No. 6,765,087), molds (e.g., Aspergillus or Trichoderma), and yeasts (e.g., Saccharomyces, Kluyveromyces, Hansenella, or Pichia (see U.S. Pat. No. 6,838,254). The production process is scalable and multi-kilogram quantities of nanobodies have been produced. Nanobodies can be formulated as a ready-to-use solution with a long shelf life. The Nanoclone method (see WO06 / 079372) is a proprietary method for generating nanobodies against desired targets based on automated high-throughput selection of B cells.

[0054] In some embodiments, the antibodies or antigen-binding fragments described herein are in the form of aptamers (see, e.g., Ellington et al., Nature. 346, 818-22, 1990; and Tuerk et al., Science. 249, 505-10, 1990, which are incorporated by reference). Examples of aptamers include nucleic acid aptamers (e.g., DNA aptamers, RNA aptamers) and peptide aptamers. Nucleic acid aptamers generally refer to nucleic acid species that are engineered through repeated rounds of in vitro selection, or equivalent methods, such as SELEX (systematic evolution of ligands by exponential enrichment), to bind to various molecular targets, such as small molecules, proteins, nucleic acids, and even cells, tissues, and organisms. See, e.g., U.S. Patent Nos. 6,376,190; and 6,387,620, which are incorporated by reference.

[0055] Peptide aptamers typically comprise a variable peptide loop attached at both ends to a protein scaffold, typically a dual structural constraint that increases the binding affinity of the peptide aptamer to levels comparable to those of antibodies (e.g., in the nanomolar range). In certain embodiments, the variable loop length may be comprised of about 10-20 amino acids (including all integers in between), and the scaffold may comprise any protein with good solubility and compaction properties. Certain exemplary embodiments utilize the bacterial protein thioredoxin-A as the scaffold protein, in which the variable loop is inserted into the reducing active site (-Cys-Gly-Pro-Cys-loop in the wild-type protein), and the two cysteine ​​side chains can form a disulfide bridge. Methods for identifying peptide aptamers are described, for example, in U.S. Patent Application Publication No. 2003 / 0108532, which is incorporated by reference. Peptide aptamer selection can be performed using different systems known in the art, including the yeast two-hybrid system.

[0056] In some embodiments, the antibodies or antigen-binding fragments described herein are in the form of avimers. Avimers refer to multimeric binding proteins or peptides engineered using in vitro exon shuffling and phage display. Multiple binding domains are linked, resulting in greater affinity and specificity compared to single epitope immunoglobulin domains. See, for example, Silverman et al., Nature Biotechnology. 23:1556-1561, 2005; U.S. Patent No. 7,166,697; and U.S. Patent Application Nos. 2004 / 0175756, 2005 / 0048512, 2005 / 0053973, 2005 / 0089932, and 2005 / 0221384, all of which are incorporated by reference.

[0057] In some embodiments, the antibodies or antigen-binding fragments described herein are in the form of adnectins. Adnectins refer to a class of targeted biologics derived from human fibronectin, an abundant extracellular protein that naturally binds to other proteins. See, for example, U.S. Patent Application Nos. 2007 / 0082365; 2008 / 0139791; and 2008 / 0220049, which are incorporated by reference. Adnectins typically consist of a natural fibronectin backbone and multiple targeting domains of specific portions of human fibronectin. The targeting domains can be engineered to enable the adnectin to specifically recognize an IL-11Rα polypeptide or an epitope thereof.

[0058] In some embodiments, the antibodies or antigen-binding fragments described herein are in the form of anticalins. Anticalins refer to a class of antibody mimics that are typically synthesized from human lipocalins, a family of binding proteins with hypervariable loop regions supported by a structurally rigid framework. See, for example, US Patent Application No. 2006 / 0058510. Anticalins typically have a size of about 20 kDa. Anticalins can be characterized by a barrel structure formed by eight antiparallel β-strands (a stable β-barrel scaffold) connected in pairs by four peptide loops and an attached α-helix. In certain aspects, conformational deviations are made in the hypervariable loop regions to achieve specific binding. See, for example, Skerra, FEBS J. 275:2677-83, 2008, incorporated by reference.

[0059] In some embodiments, the antibodies or antigen-binding fragments described herein are in the form of designed ankyrin repeat proteins (DARPins). DARPins comprise a class of non-immunoglobulin proteins that may offer advantages over antibodies for target binding in drug discovery and drug development. Among other applications, DARPins are ideally suited for in vivo imaging or delivery of toxins or other therapeutic payloads due to favorable molecular properties, including small size and high stability. The low-cost production in bacteria and rapid generation of many target-specific DARPins makes the DARPin approach useful for drug discovery. In addition, DARPins can be easily produced in multispecific formats, offering the potential to target effector DARPins to specific organs or to target multiple receptors with one molecule composed of several DARPins. See, e.g., Stumpp et al., Curr Opin Drug Discov Devel. 10:153-159, 2007; U.S. Patent Application No. 2009 / 0082274; and PCT / EP2001 / 10454, which are incorporated by reference.

[0060] Also included are heavy chain dimers, such as antibodies from camelids and sharks. Camelid and shark antibodies contain two homodimeric pairs of V-like and C-like domains (neither of which have light chains). The V of IgG heavy chain dimers in camelids H Since the heavy chains do not have to make hydrophobic interactions with the light chains, the regions in the heavy chains that normally contact the light chains are changed to hydrophilic amino acid residues in camelids. The VH domains of the heavy chain dimer IgG are called VHH domains. Shark Ig-NAR contains a homodimer of one variable domain (called the V-NAR domain) and five C-like constant domains (C-NAR domains).

[0061] In camelids, the diversity of the antibody repertoire is determined by complementarity determining regions (CDRs) 1, 2, and 3 in the VH or VHH regions. CDR3 in camelid VHH regions is characterized by its relatively long length, averaging 16 amino acids (Muyldermans et al., 1994, Protein Engineering 7(9): 1129). This is in contrast to the CDR3 regions of antibodies of many other species. For example, the CDR3 of mouse VH has an average of 9 amino acids. Libraries of camelid-derived antibody variable regions maintain the in vivo diversity of camelid variable regions and can be made, for example, by the methods disclosed in U.S. Patent Application No. 20050037421 published February 17, 2005.

[0062] In certain embodiments, the antibody or antigen-binding fragment thereof is humanized. These embodiments generally refer to chimeric molecules, prepared using recombinant techniques, having an antigen-binding site derived from an immunoglobulin from a non-human species and the remaining immunoglobulin structure of the molecule based on the structure and / or sequence of a human immunoglobulin. The antigen-binding site may comprise either a complete variable domain fused onto a constant domain, or only the CDRs (whole or part) grafted onto suitable framework regions in the variable domain. The epitope-binding site may be wild-type or modified by one or more amino acid substitutions. This removes the constant region as an immunogen in human individuals, but still leaves the possibility of an immune response against the foreign variable region (LoBuglio et al., PNAS USA 86:4220-4224, 1989; Queen et al., PNAS USA. 86:10029-10033, 1988; Riechmann et al., Nature. 332:323-327, 1988). Exemplary methods for antibody humanization include those described in US Pat. No. 7,462,697.

[0063] Another approach focuses not only on providing constant regions of human origin, but also on modifying the variable regions and reshaping them as closely as possible to human form. Both heavy and light chain variable regions are known to contain three complementarity determining regions (CDRs) that are relatively conserved in a given species and flanked by four framework regions (FRs) that presumptively provide a scaffold for the CDRs, and that vary in response to the epitope in question and determine the binding ability. When a non-human antibody is prepared for a specific epitope, the variable region can be reshaped or humanized by grafting the CDRs derived from the non-human antibody onto the FRs present in the modified human antibody. Application of this approach to a variety of antibodies has been reported in Sato et al., Cancer Res. 53:851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering. 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA. 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA. 88:2869-2873, 1991; Carter et al., PNAS USA. 89:4285-4289, 1992; and Co et al., J Immunol. 148:1149-1154, 1992. In some embodiments, a humanized antibody retains all CDR sequences (e.g., a humanized mouse antibody that contains all six CDRs from the mouse antibody). In some embodiments, only a portion of the CDR sequences are grafted from a non-human antibody (Bowers et al., J. Biol. Chem. 288:7688-7696, 2013).In certain embodiments, a humanized antibody has one or more CDRs (one, two, three, four, five, six) that are altered with respect to an original antibody, which are also referred to as one or more CDRs "derived from" one or more CDRs from the original antibody.

[0064] In certain embodiments, the antibody is a chimeric antibody. In this regard, a chimeric antibody is composed of an antigen-binding fragment of an antibody operatively linked or otherwise fused to a heterologous Fc portion of a different antibody. In certain embodiments, the Fc domain or heterologous Fc domain is of human origin. In certain embodiments, the Fc domain or heterologous Fc domain is of murine origin. In other embodiments, the heterologous Fc domain may be from a different Ig class than the parent antibody, including IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), and IgM. In further embodiments, the heterologous Fc domain may be composed of CH2 and CH3 domains from one or more of the different Ig classes. As described above for humanized antibodies, an antigen-binding fragment of a chimeric antibody may include only one or more of the CDRs of an antibody described herein (e.g., one, two, three, four, five, or six CDRs of an antibody described herein) or may include the entire variable domain (VL, VH, or both).

[0065] The term "bond" refers to a direct association between two molecules, for example, due to covalent, electrostatic, hydrophobic, and ionic and / or hydrogen bonding interactions, including, for example, salt bridges and water bridges.

[0066] By "coding sequence" is meant any nucleic acid sequence that contributes to the code for the polypeptide product of a gene. In contrast, the term "non-coding sequence" refers to any nucleic acid sequence that does not directly contribute to the code for the polypeptide product of a gene.

[0067] Throughout this specification, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer, or group of elements or integers, but not the exclusion of any other element or integer, or group of elements or integers.

[0068] "Consisting of" is meant to include, but is not limited to, everything that follows the phrase "consisting of." Thus, the phrase "consisting of" indicates that the recited elements are required or essential, and that no other elements will be present. By "consisting essentially of," it is meant to include any elements recited after the phrase, and to be limited to other elements that do not interfere with or contribute to the activity or action specified in this disclosure for the recited elements. Thus, the phrase "consisting essentially of" indicates that the recited elements are required or essential, but that other elements are optional and may or may not be present depending on whether they substantially affect the activity or action of the recited elements.

[0069] The term "effector function" in the context of an antibody, or "ADCC effector function", refers to the ability of that antibody to associate with other arms of the immune system, including, for example, activation of the classical complement pathway or through the association of Fc receptors. The complement-dependent pathway is driven primarily by the interaction of C1q with the C1 complex and clustered antibody Fc domains. Antibody-dependent cellular cytotoxicity (ADCC) is driven primarily by the interaction of Fc receptors (FcRs) on the surface of effector cells (natural killer cells, macrophages, monocytes, and eosinophils) that bind to the Fc region of IgG, which itself is bound to the target cell. Fc receptors (FcRs) are key immunoregulatory receptors that connect the antibody-mediated (humoral) immune response to cellular effector functions. Receptors for all classes of immunoglobulins have been identified and include FcγR (IgG), FcεRI (IgE), FcαRI (IgA), FcμR (IgM), and FcδR (IgD). There are at least three receptor classes for human IgG found on leukocytes: CD64 (FcγRI), CD32 (FcγRIIa, FcγRIIb, and FcγRIIc), and CD16 (FcγRIIIa and FcγRIIIb). FcγRI is a high affinity receptor (nanomolar range K D ), whereas FcγRII and FcγRIII have low to intermediate affinity (micromolar range K D Upon Fc binding, signaling pathways are triggered that result in the secretion of various substances, such as lytic enzymes, perforin, granzymes, and tumor necrosis factor, which mediate the destruction of the target cell. The level of ADCC effector function varies for human IgG subtypes. This is dependent on the allotype and the specific FcvR, but in simple terms, ADCC effector function is "high" for human IgG1 and IgG3, and "low" for IgG2 and IgG4.

[0070] The terms "endotoxin-free" or "substantially endotoxin-free" generally refer to compositions, solvents, and / or containers that contain at most trace amounts of endotoxin (e.g., amounts that have no clinically adverse physiological effects on a subject), and preferably undetectable amounts of endotoxin. Endotoxins are toxins associated with certain microorganisms, such as bacteria, typically gram-negative bacteria, although endotoxins can be found in gram-positive bacteria, such as Listeria monocytogenes. The most common endotoxins are lipopolysaccharides (LPS) or lipooligosaccharides (LOS), found in the outer membrane of various gram-negative bacteria, which represent a central pathogenic feature in the ability of these bacteria to cause disease. Small amounts of endotoxin in humans can produce fever, a drop in blood pressure, and activation of inflammation and coagulation, among other adverse physiological effects.

[0071] Thus, in pharmaceutical production, it is often desirable to remove most or all traces of endotoxin from the drug product and / or drug container, since even small amounts can cause adverse effects in humans. Dehydrogenation ovens can be used for this purpose, since temperatures above 300°C are typically required to break down most endotoxins. For example, based on primary packaging materials, such as syringes or vials, a combination of a glass temperature of 250°C and a holding time of 30 minutes is often sufficient to achieve a 3 log reduction in endotoxin levels. Other methods of removing endotoxins are contemplated, including, for example, chromatography and filtration methods described herein and known in the art.

[0072] Endotoxin can be detected using routine techniques known in the art. For example, the Limulus amebocyte lysis assay utilizes blood from horseshoe crabs and is a highly sensitive assay for detecting the presence of endotoxin. In this test, very low levels of LPS can cause detectable clotting of the Limulus lysate due to a powerful enzyme cascade that amplifies this reaction. Endotoxin can also be quantified by enzyme-linked immunosorbent assay (ELISA). To be substantially endotoxin-free, endotoxin levels can be less than about 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.08, 0.09, 0.1, 0.5, 1.0, 1.5, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, or 10 EU / mg of active compound. Typically, 1 ng of lipopolysaccharide (LPS) corresponds to about 1-10 EU.

[0073] The term "epitope" includes any determinant, preferably a polypeptide determinant, capable of specific binding to an immunoglobulin or T-cell receptor. An epitope includes the region of an antigen that is bound by an antibody. In certain embodiments, epitopic determinants include chemically active surface groupings of molecules, such as amino acids, sugar side chains, phosphoryls, or sulfonyls, and in certain embodiments may have specific three-dimensional structural characteristics, and / or specific charge characteristics. Epitopes may be contiguous or discontinuous with respect to the primary structure of an antigen, e.g., an IL-11Rα polypeptide. In certain embodiments, an epitope comprises, consists of, or consists essentially of about, at least about, or no more than about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous amino acids (i.e., a linear epitope) or non-contiguous amino acids (i.e., a conformational epitope) of a reference sequence (see, e.g., Table B1) or target molecule described herein.

[0074] An "epitope" includes that portion of an antigen or other macromolecule capable of forming a binding interaction that interacts with the variable region binding pocket of a binding protein. Such a binding interaction may be manifested as an intermolecular contact with one or more amino acid residues of a CDR. Antigen binding may involve a CDR3 or a CDR3 pair. An epitope may be a linear peptide sequence (i.e., "continuous") or may be composed of a non-contiguous amino acid sequence (i.e., "conformational" or "discontinuous"). A binding protein may recognize one or more amino acid sequences; thus, an epitope may define more than one distinct amino acid sequence. Epitopes recognized by a binding protein may be determined by peptide mapping and sequence analysis techniques well known to those skilled in the art. A "cryptic epitope" or "cryptic binding site" is an epitope or binding site of a protein sequence that is not exposed or is substantially protected from recognition in an unmodified polypeptide, but is capable of being recognized by a binding protein of a denatured or proteolytic polypeptide. An amino acid sequence that is not exposed or only partially exposed in the unmodified polypeptide structure is a potential cryptic epitope. If an epitope is not exposed or only partially exposed, then it is likely to be buried within the interior of the polypeptide. Candidate cryptic epitopes can be identified, for example, by examining the three-dimensional structure of the unmodified polypeptide.

[0075] The term "half maximum effective concentration" or "EC 50 " refers to the concentration of an agent (e.g., an antibody) described herein that induces a response halfway between baseline and maximum after some specified exposure time; EC 50 Thus, "EC50" represents the concentration of a compound at which 50% of its maximum effect is observed. EC50 also represents the plasma concentration required to obtain 50% of the maximum effect in vivo. Similarly, "EC 90 "EC" refers to the concentration of a drug or composition at which 90% of its maximum effect is observed. 90 " is "E.C. 50" and the Hill slope, or it can be determined directly from the data using routine knowledge in the art. In some embodiments, the EC 50 is less than about 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, or 500 nM. In some embodiments, the agent has an EC 5 It has a 0 value.

[0076] "Immune response" refers to any immunological response derived from the immune system, including responses from the cellular and cellular, innate and adaptive immune systems. Exemplary cellular immune cells include, for example, lymphocytes, macrophages, T cells, B cells, NK cells, neutrophils, eosinophils, dendritic cells, mast cells, monocytes, and all subsets thereof. Cellular responses include, for example, effector functions, cytokine release, phagocytosis, efferocytosis, translocation, trafficking, proliferation, differentiation, activation, suppression, cell-cell interactions, apoptosis, and the like. Humoral responses include, for example, IgG, IgM, IgA, IgE, responses and their corresponding effector functions.

[0077] The "half-life" of an agent, such as an antibody, can refer to the time it takes for the agent to lose half of its pharmacological, physiological, or other activity, relative to such activity upon administration to the serum or tissues of an organism, or relative to any other defined time point. "Half-life" can also refer to the time it takes for the amount or concentration of the agent to fall by half of the starting amount administered to the serum or tissues of an organism, relative to such amount or concentration upon administration to the serum or tissues of an organism, or relative to any other defined time point. Half-life can be measured in serum and / or in any one or more selected tissues.

[0078] The terms "modulating" and "altering" include "increasing," "enhancing," or "stimulating," as well as "decreasing," "reducing," or "inhibiting," typically in a statistically significant or physiologically significant amount or degree relative to a control. An "increased," "stimulated," or "enhanced" amount is typically a "statistically significant" amount and can include an increase that is 1.1, 1.2, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100-fold or more (e.g., 500, 1000-fold) (including all integers and ranges therebetween, e.g., 1.5, 1.6, 1.7, 1.8) than the amount produced without the composition (e.g., absence of agent) or by a control composition. A "decreased" or "reduced" or "inhibited" amount is typically a "statistically significant" amount and can include a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% decrease (including all integers and ranges therebetween) in the amount produced by no composition (e.g., absence of agent) or a control composition. Examples of comparisons and "statistically significant" amounts are described herein.

[0079] The terms "polypeptide", "protein" and "peptide" are used interchangeably and refer to a polymer of amino acids, not limited to any particular length. The term "enzyme" includes polypeptide or protein catalysts. This term includes modifications, such as myristoylation, sulfation, glycosylation, phosphorylation, and the addition or deletion of a signal sequence. The term "polypeptide" or "protein" refers to one or more chains of amino acids, where each chain includes amino acids covalently linked by peptide bonds, where the polypeptide or protein can include multiple chains non-covalently and / or covalently linked together by peptide bonds, including naturally occurring proteins, i.e., proteins produced by natural cells and specific non-recombinant cells, or genetically engineered or recombinant cells, and includes molecules having the amino acid sequence of a naturally occurring protein, or molecules having deletions from, additions to, and / or substitutions of one or more amino acids of the naturally occurring sequence. In certain embodiments, a polypeptide is a "recombinant" polypeptide produced by a recombinant cell that contains one or more recombinant DNA molecules, which are typically made with heterologous polynucleotide sequences, or combinations of polynucleotide sequences not otherwise found in the cell.

[0080] The terms "polynucleotide" and "nucleic acid" include mRNA, RNA, cRNA, cDNA, and DNA. The terms refer to polymeric forms of nucleotides, typically at least 10 bases in length, either ribonucleotides or deoxynucleotides, or modified forms of either type of nucleotide. The terms include single-stranded and double-stranded forms of DNA. The terms "isolated DNA" and "isolated polynucleotide" and "isolated nucleic acid" refer to molecules that are isolated free of total genomic DNA of a particular species. Thus, an isolated DNA segment that encodes a polypeptide refers to a DNA segment that contains one or more coding sequences that is isolated substantially apart from or purified free of total genomic DNA of the species from which the DNA segment is obtained. Also included are non-coding polynucleotides (e.g., primers, probes, oligonucleotides), which do not encode a polypeptide. Also included are recombinant vectors, including, for example, expression vectors, viral vectors, plasmids, cosmids, phagemids, phages, viruses, and the like.

[0081] Additional coding or non-coding sequences may, but need not be, present within the polynucleotides described herein, and the polynucleotides may, but need not be linked to other molecules and / or supporting materials. Thus, a polynucleotide or expressible polynucleotide, regardless of the length of the coding sequence itself, may be combined with other sequences, for example, expression control sequences.

[0082] "Expression control sequences" include regulatory sequences of nucleic acids or corresponding amino acids, such as promoters, leaders, enhancers, introns, recognition motifs for RNA or DNA binding proteins, polyadenylation signals, terminators, internal ribosome entry sites (IRES), secretion signals, subcellular localization signals, and the like, which are capable of influencing the transcription or translation of coding sequences in a host cell, or the intracellular or cellular location. Exemplary expression control sequences are described in Goeddel; Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990).

[0083] A "promoter" is a DNA regulatory region capable of binding RNA polymerase in a cell and initiating transcription of a downstream (3' direction) coding sequence. As used herein, a promoter sequence is bounded at its 3' end by a transcription initiation site, extends upstream (5' direction), and includes the minimum number of bases or elements required to initiate transcription at a detectable level above background. A transcription initiation site (conveniently defined by mapping with nuclease S1) can be found within the promoter sequence, as well as protein binding domains (consensus sequences) responsible for binding RNA polymerase. Eukaryotic promoters often, but not always, contain "TATA" and "CAT" boxes. Prokaryotic promoters contain Shine-Dalgarno sequences in addition to the -10 and -35 consensus sequences.

[0084] Numerous promoters, including constitutive, inducible, and repressible promoters from a variety of different sources, are well known in the art. Representative sources include, for example, viral, mammalian, insect, plant, yeast, and bacterial cell types), and suitable promoters from these sources are readily available online or based on publicly available sequences, for example, from repositories, such as ATCC, as well as other commercial or individual sources, or can be synthetically produced. Promoters can be unidirectional (i.e., initiate transcription in one direction) or bidirectional (i.e., initiate transcription in either the 3' or 5' direction). Non-limiting examples of promoters include, for example, the T7 bacterial expression system, the pBAD(araA) bacterial expression system, the cytomegalovirus (CMV) promoter, the SV40 promoter, and the RSV promoter. Inducible promoters include the Tet system (U.S. Pat. Nos. 5,464,758 and 5,814,618), the ecdysone-inducible system (No et al., Proc. Natl. Acad. Sci. (1996) 93(8):3346-3351; the T-REx™ system (Invitrogen, Carlsbad, Calif.), LacSwitch® (Stratagene, San Diego, Calif.), and the Cre-ERT tamoxifen-inducible recombinase system (Indra et al. Nuc. Acid. Res. (1999) 27 (22): 4324-4327; Nuc. Acid. Res. (2000) 28 (23): e99; U.S. Pat. No. 7,112,715; and Kramer & Fussenegger, Methods Mol. Biol. (2005) 308: 123-144), or any promoter known in the art appropriate for expression in the desired cells.

[0085] An "expressible polynucleotide" includes a cDNA, RNA, mRNA, or other polynucleotide that contains at least one coding sequence and, optionally, at least one expression control sequence, e.g., a transcriptional and / or translational regulatory element, and is capable of expressing an encoded polypeptide upon introduction into a cell, e.g., a cell in a subject.

[0086] Various viral vectors that can be utilized to deliver expressible polynucleotides include adenoviral vectors, herpes virus vectors, vaccinia virus vectors, adeno-associated virus (AAV) vectors, and retroviral vectors. In some cases, the retroviral vector is a derivative of a murine or avian retrovirus, or is a lentiviral vector. Examples of retroviral vectors that can insert a single foreign gene include, but are not limited to, Moloney murine leukemia virus (MoMuLV), Harvey murine sarcoma virus (HaMuSV), mouse mammary tumor virus (MuMTV), SIV, BIV, HIV, and Rous sarcoma virus (RSV). Many additional retroviral vectors can incorporate multiple genes. All of these vectors can transfer or incorporate genes for selectable markers, allowing transduced cells to be identified and generated. The polypeptide sequence of interest can be inserted into the viral vector along with another gene that codes for a ligand for a receptor on a specific target cell, making the vector target specific, for example. Retroviral vectors can be made target specific, for example, by inserting a polynucleotide encoding a protein. An exemplary targeting can be achieved by targeting retroviral vectors using an antibody. Those skilled in the art will know, or can easily ascertain, without undue experimentation, specific polynucleotide sequences that can be inserted into a retroviral genome to allow target specific delivery of retroviral vectors.

[0087] In certain embodiments, the expressible polynucleotide is a modified RNA or modified mRNA polynucleotide, e.g., a non-natural RNA analog. In certain embodiments, the modified RNA or mRNA polypeptide comprises one or more modified or non-natural bases, e.g., nucleotide bases other than adenine (A), guanine (G), cytosine (C), thymine (T), and / or uracil (U). In some embodiments, the modified mRNA comprises one or more modified or non-natural internucleotide linkages. Expressible RNA polynucleotides for delivering encoded therapeutic polypeptides are described, for example, in Kormann et al., Nat Biotechnol. 29:154-7, 2011; and U.S. Patent Application Nos. 2015 / 0111248; 2014 / 0243399; 2014 / 0147454; and 2013 / 0245104, which are incorporated by reference in their entireties.

[0088] The term "isolated" polypeptide or protein as referred to herein means that the subject protein (1) is free from at least some other proteins with which it would typically be found in nature, (2) is essentially free from other proteins from the same source, e.g., from the same species, (3) is expressed by cells from a different species, (4) is separated from at least about 50 percent of the polynucleotides, lipids, carbohydrates, or other materials with which it is naturally associated, (5) is not associated (by covalent or non-covalent interactions) with portions of proteins with which it is naturally associated, (6) is operatively associated (by covalent or non-covalent interactions) with polypeptides with which it is not naturally associated, or (7) does not occur in nature. Such isolated proteins may be encoded by genomic DNA, cDNA, mRNA, or other RNA, may be of synthetic origin, or any combination thereof. In certain embodiments, an isolated protein is substantially free of proteins or polypeptides found in its natural environment, or other contaminants that would interfere with its use (therapeutic, diagnostic, preventative, research, or otherwise).

[0089] In certain embodiments, the purity of any given agent (e.g., an antibody) in a composition may be defined. For example, a particular composition may include an agent that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% pure on a protein basis or weight-by-weight basis, including, for example, all decimal points measured and ranges therebetween, and is not limited by any means by high performance liquid chromatography (HPLC), a well-known form of column chromatography frequently used in biochemistry and analytical chemistry to separate, identify, and quantify compounds.

[0090] The term "reference sequence" generally refers to a nucleic acid coding sequence, or amino acid sequence, to which another sequence is compared. All polypeptide and polynucleotide sequences described herein are included as reference sequences, including those described by name and those described in the Tables and Sequence Listing.

[0091] Certain embodiments include biologically active "variants" and "fragments" of the polypeptides (e.g., antibodies) described herein, as well as polynucleotides encoding the same. "Variants" include one or more substitutions, additions, deletions, and / or insertions compared to a reference polypeptide or polynucleotide (see, e.g., Tables and Sequence Listing). A variant polypeptide or polynucleotide comprises an amino acid or nucleotide sequence with at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity or similarity or homology to a reference sequence, as described herein, and substantially retains an activity of the reference sequence. Also included are sequences that consist of a reference sequence or that differ from a reference sequence by additions, deletions, insertions, or substitutions of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150 or more amino acids or nucleotides, and that substantially retain the activity of the reference sequence. In certain embodiments, the additions or deletions include C-terminal and / or N-terminal additions and / or deletions.

[0092] The term "sequence identity" or, as used herein, includes, for example, "at least 50% identical sequences", refers to the degree to which sequences are identical on a nucleotide-by-nucleotide or amino acid-by-amino acid basis over a window of comparison. Thus, "percentage of sequence identity" may be calculated by comparing two optimally aligned sequences over a window of comparison, determining the number of positions at which identical nucleobases (e.g., A, T, C, G, I) or identical amino acid residues (e.g., Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cys, and Met) occur in both sequences, resulting in the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison (i.e., the window size), and multiplying the result by 100 to result in the percentage of sequence identity. Optimal alignment of sequences to align the comparison window may be performed by computerized implementation of algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package Release 7.0, Genetics Computer Group, 575 Science Drive Madison, Wis., USA) or by inspection and best alignment (i.e., resulting in the highest percentage homology over the comparison window) generated by any of the various methods selected. Reference may also be made to the BLAST family of programs, as disclosed, for example, by Altschul et al., Nucl. Acids Res. 25:3389, 1997.

[0093] The term "solubility" refers to the ability of an agent (e.g., an antibody) provided herein to dissolve in a liquid solvent to form a homogenous solution. Solubility is typically expressed as a concentration by either mass of solute per unit volume of solvent (e.g., g of solute per kg of solvent, g per dL (100 mL), mg / ml, etc.), molar concentration, molar concentration, molar fraction, or other similar descriptions of concentration. The maximum equilibrium amount of solute that can be dissolved per volume of solvent is the solubility of the solute in that solvent under specific conditions, including temperature, pressure, pH, and the nature of the solvent. In certain embodiments, solubility is measured at physiological pH or other pHs, such as pH 5.0, pH 6.0, pH 7.0, pH 7.4, pH 7.6, pH 7.8, or pH 8.0 (e.g., about pH 5-8). In certain embodiments, solubility is measured in water or a physiological buffer, such as PBS or NaCl (NaPO 4 In certain embodiments, solubility is measured at relatively low pH (e.g., pH 6.0) and relatively high salt (e.g., 500 mM NaCl and 10 mM NaPO 4 ) in a biological fluid (solvent), such as blood or serum. In certain embodiments, the temperature can be about room temperature (e.g., about 20, 21, 22, 23, 24, 25° C.) or about body temperature (37° C.). In certain embodiments, the agent has a solubility of at least about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 40, 50, 60, 70, 80, 90, or 100 mg / ml at room temperature or at 37° C.

[0094] A "subject" or "subject in need thereof" or a "patient" or "patient in need thereof" includes mammalian subjects, such as human subjects.

[0095] "Substantially" or "essentially" means nearly entirely or completely, for example, 95%, 96%, 97%, 98%, 99%, or more of a given amount of a portion.

[0096] By "statistically significant", it means that the result is unlikely to have occurred by chance. Statistical significance can be determined by any method known in the art. A commonly used measure of significance includes p-value, which is the frequency or probability that the observed event would occur if the null hypothesis is true. If the p-value obtained is less than the significance level, then the null hypothesis is rejected. In simple cases, the significance level is defined as a p-value of 0.05 or less.

[0097] "Therapeutic response" refers to an improvement in symptoms (whether sustained or not) following administration of one or more therapeutic agents.

[0098] As used herein, the terms "therapeutically effective amount," "therapeutic dose," "prophylactically effective amount," or "diagnostically effective amount" are the amount of an agent (e.g., an anti-IL-11Rα antibody, an immunotherapeutic agent) required to elicit a desired biological response following administration.

[0099] As used herein, "treatment" of a subject (e.g., a mammal, such as a human) or cell is any type of intervention used in an attempt to change the natural course of an individual or cell. Treatment includes, but is not limited to, administration of a pharmaceutical composition and can be performed either prophylactically or following the initiation of a pathological event or contact with a pathogenic agent. Also included are prophylactic treatments, which can be directed to reducing the rate of progression of the disease or condition being treated, delaying the onset of the disease or condition, or reducing the severity of its onset. "Treatment" or "prevention" does not necessarily indicate a complete eradication, cure, or prevention of the disease or condition, or its associated symptoms.

[0100] The term "wild-type" refers to a gene or gene product (eg, a polypeptide) that is most frequently observed in a population and thus arbitrarily designed as the "normal" or "wild-type" form of the gene.

[0101] Each embodiment in this specification should be applied mutatis mutandis to all other embodiments unless expressly stated otherwise.

[0102] Anti-IL-11Rα antibody Certain embodiments include antibodies, and antigen-binding fragments thereof, that bind to interleukin-11 receptor subunit alpha (IL-11Rα). In some embodiments, the antibodies or antigen-binding fragments thereof modulate (e.g., interfere with, antagonize, inhibit) the binding of IL-11Rα to its ligand, interleukin-11 (IL-11). In certain embodiments, the antibodies or antigen-binding fragments thereof bind to the complementarity determining region V H CDR1, V H CDR2, and V H The heavy chain variable region (V H ), and complementarity determining region V L CDR1, V L CDR2, and V L The light chain variable region (V L ) H Array, V H CDR1 sequence, V H CDR2 sequence, V H CDR3 sequence, V L Array, V L CDR1 sequence, V L CDR2 sequence, and V L The CDR3 sequences are provided in Tables A1 and A2 below. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5]

[0103] Thus, in certain embodiments, the antibody or antigen-binding fragment thereof comprises: Complementarity determining region V selected from Table A1 H CDR1, V H CDR2, and V H V containing the CDR3 sequence H and a complementarity determining region V selected from Table A1, L CDR1, V L CDR2, and V L V containing the CDR3 sequence L The sequence, as well as variants thereof that bind to IL-11Rα.

[0104] In certain embodiments, the CDR sequences are as follows: V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 1 to 3, respectively, and V L CDR1, V L CDR2, and V L the CDR3 sequences include SEQ ID NOs: 4 to 6, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 7 to 9, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 10 to 12, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 13 to 15, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 16 to 18, respectively; V H CDR1, V H CDR2, and V HThe CDR3 sequences include SEQ ID NOs: 19 to 21, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 22 to 24, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 25 to 27, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 28 to 30, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 31 to 33, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 34 to 36, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 37 to 39, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 40 to 42, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 43 to 45, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 46 to 48, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 49 to 51, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 52 to 54, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 55 to 57, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 58 to 60, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 61 to 63, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 64 to 66, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 67 to 69, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 70 to 72, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 73 to 75, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 76 to 78, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 79 to 81, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 82 to 84, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 85 to 87, respectively, and V L CDR1, VL CDR2, and V L CDR3 sequences include SEQ ID NOs: 88 to 90, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 91 to 93, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 94 to 96, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 97 to 99, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 100 to 102, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 103 to 105, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 106 to 108, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 109 to 111, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 112-114, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 115 to 117, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 118-120, respectively; V H CDR1, V HCDR2, and V H The CDR3 sequences include SEQ ID NOs: 121 to 123, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 124 to 126, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 127 to 129, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 130 to 132, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 133 to 135, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 136 to 138, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 139 to 141, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 142 to 144, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 145 to 147, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 148 to 150, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 151 to 153, respectively, and V L CDR1, V L CDR2, and VL CDR3 sequences include SEQ ID NOs: 154 to 156, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 157 to 159, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 160 to 162, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 163 to 165, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 166-168, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 169 to 171, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 172 to 174, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 175 to 177, respectively, and V L CDR1, V L CDR2, and V L CDR3 sequences include SEQ ID NOs: 178 to 180, respectively; V H CDR1, V H CDR2, and V H The CDR3 sequences include SEQ ID NOs: 181 to 183, respectively, and V L CDR1, V L CDR2, and V L the CDR3 sequences comprise SEQ ID NOs: 184 to 186, respectively; or V H CDR1, V H CDR2, and VH The CDR3 sequences include SEQ ID NOs: 187 to 189, respectively, and V L CDR1, V L CDR2, and V L The CDR3 sequences include SEQ ID NOs: 190 to 192, respectively.

[0105] Also included are minor variants of the aforementioned CDRs. Exemplary variants bind IL-11Rα and include any one or more of the individual CDRs, e.g., the V and Vs described herein. H CDR1, V H CDR2, V H CDR3, V L CDR1, V L CDR2 and / or V L Have a total of 1, 2, or 3 changes in any one or more of the CDR3 sequences. Exemplary "changes" include amino acid substitutions, additions, and deletions.

[0106] Exemplary V H and V L The sequences are provided in Table A2 below. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8]

[0107] Thus, in certain embodiments, the antibody, or antigen-binding fragment thereof, binds to IL-11Rα and is selected from Table A2. H Sequence and corresponding V L In certain embodiments, the V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, e.g., in which V H In some embodiments, V has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in one or more framework regions. L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to a sequence selected from Table A2, e.g., in which V L has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in one or more framework regions.

[0108] In some embodiments, the V of the antibody or antigen-binding fragment H and V L is as follows: V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 193, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 194; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 195, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 196; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 197, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 198; VH comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 199, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 200; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 201, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:202; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 203, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:204; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 205, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:206; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 207, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:208; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 209, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:210; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 211, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:212; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 213, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:214; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 215, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:216; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 217, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:218; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 219, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 220; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 221, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:222; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 223, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:224; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 225, and Lcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:226; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 227, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:228; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 229, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:230; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 231, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:232; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 233, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:234; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 235, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:236; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 237, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:238; V Hcomprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 239, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:240; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 241, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:242; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 243, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:244; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 245, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:246; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 247, and V L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:248; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 249, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 250; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 251, and L comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:252; V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 253, and L comprises a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 254; or V H comprises a sequence at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO: 255, and L comprises a sequence that is at least 80, 85, 90, 95, 97, 98, 99, or 100% identical to SEQ ID NO:256.

[0109] Also included are variants thereof that bind to IL-Rα, such as the aforementioned V H and / or V L A variant having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in any one or more framework regions of the sequence. Exemplary "changes" include amino acid substitutions, additions, and deletions.

[0110] As discussed above, the antibodies or antigen-binding fragments thereof described herein bind to IL-11Rα, e.g., membrane-bound IL-11Rα. In certain embodiments, the antibodies or antigen-binding fragments thereof bind to human IL-11Rα, or a region or fragment thereof. The amino acid sequence of human IL-11Rα is provided in Table B1 below. [Table 3]

[0111] Thus, in certain embodiments, the antibody or antigen-binding fragment thereof binds to an IL-11Rα sequence in Table B1, e.g., a human IL-11Rα sequence. In certain embodiments, the antibody or antigen-binding fragment thereof binds to a fibronectin type III domain in human IL-11Rα, e.g., consisting of about residues 112-219 of SEQ ID NO:257.

[0112] In some embodiments, the antibody or antigen-binding fragment thereof may be administered with a binding affinity of about 1 pM to about 10 pM to about 500 pM, or about, at least about, or less than about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 300, 400, or 500 pM, or optionally, about 1 pM to about 500 pM, about 1 pM ~about 400pM, about 1pM to about 300pM, about 1pM to about 200pM, about 1pM to about 100pM, about 1pM to about 50pM, about 1pM to about 40pM, about 1pM to about 30pM, about 1pM to about 20pM, about 1pM Approximately 10 pM, approximately 1 pM to approximately 5 pM, approximately 5 pM to approximately 500 pM, approximately 5 pM to approximately 400 pM, approximately 5 pM to approximately 300 pM, approximately 5 pM to approximately 200 pM, approximately 5 pM to approximately 100 pM, approximately 5 pM to approximately 50 pM, approximately 5 pM to approximately 40 pM, about 5 pM to about 30 pM, about 5 pM to about 20 pM, about 5 pM to about 10 pM, about 10 pM to about 500 pM, about 10 pM to about 400 pM, about 10 pM to about 300 pM, about 10 pM to about 200 pM, about 10 pM to about 100 pM, about 10 pM to about 50 pM, about 10 pM to about 40 pM, about 10 pM to about 30 pM, about 10 pM to about 20 pM, or about 20 pM to about 500 pM, about 20 pM to about 400 pM, about 20 pM to about 300 pM, about 20 pM to about 200 pM, about 20 pM to about 100 pM, about 20 pM to about 50 pM, about 20 pM to about 40 pM, about 20 pM to about 30 pM, or about 30 pM to about 500 pM, about 30 pM to about 400 pM, about 30 pM to about 300 pM, about 30 pM to about 200 pM, about 30 pM to about 100 pM, about 30 pM to about 50 pM, or about 30 pM to about 40 pM. In certain embodiments, the antibody or antigen-binding fragment thereof has increased binding affinity for human IL-11Rα compared to that of the TS7 antibody and the 8E2 antibody (see, e.g., U.S. Patent Nos. 9,796,782; 9,340,618).

[0113] In some embodiments, the antibody, or antigen-binding fragment thereof, is an IL-11Rα antagonist. In some examples, the antibody, or antigen-binding fragment thereof, antagonizes the binding and / or signaling activity between IL-11Rα and its ligand IL-11. In some embodiments, the antibody, or antigen-binding fragment thereof, antagonizes or reduces the binding and / or signaling activity between IL-11Rα and IL-11 by about or at least about 10-1000% (e.g., about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000% or more), e.g., in a cell-based assay. In some embodiments, the anti-IL-11Rα antibody, or antigen-binding fragment thereof, reduces IL-11-mediated STAT3 phosphorylation. In an exemplary assay, cells expressing IL-11Rα and gp130 are cultured in the presence of IL-11 in the presence or absence of IL-11Rα binding protein. The level of STAT3 phosphorylation is then assessed by Western blotting or FACS using an antibody specific for phosphorylated STAT3. An exemplary assay using FACS is described in Dams-Kozlowska et al., BMC Biotechnol, 12:8, 2012. In certain embodiments, the antibody or antigen-binding fragment thereof has increased potency as an IL-11 signaling antagonist compared to that of the TS7 and 8E2 antibodies (see, e.g., U.S. Patent Nos. 9,796,782; 9,340,618). In certain embodiments, the antibody or antigen-binding fragment thereof has no detectable agonist activity with respect to IL-11 signaling.

[0114] In some embodiments, the antibody, or antigen-binding fragment thereof, inhibits or otherwise reduces IL-11Rα dimerization or complex formation, e.g., with gp130. In certain embodiments, the antibody, or antigen-binding fragment thereof, inhibits or otherwise reduces IL-11Rα dimerization or complex formation by about or at least about 10-1000% (e.g., about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000% or more), e.g., in a cell-based assay.

[0115] In some embodiments, the anti-IL-11Rα antibody or antigen-binding fragment thereof reduces proliferation of cells expressing IL-11Rα and gp130 (e.g., BaF3 cells, B9 cells, T10 cells) (e.g., cells that naturally express or have been engineered to express both proteins) cultured in the presence of IL-11. Methods for assessing cell proliferation are known in the art and include, for example, MTT reduction and / or thymidine incorporation. Assays using B9 or T10 cells have been described (see Dams-Kozlowska et al., BMC Biotechnol, 12: 8, 2012; and Yokote et al., J AOAC, 83:1053-1057, 2000). For T10 cells, proliferation can be measured by colorimetrically detecting a decrease in the tetrazolium compound, 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzenedisulfonate (WST-1). An IL-11Rα binding protein that reduces the level of proliferation compared to that observed in the absence of the IL-11Rα binding protein is believed to reduce or otherwise reduce IL-11R signaling.

[0116] In some embodiments, the anti-IL-11Rα antibody or antigen-binding fragment thereof reduces IL-11-mediated proliferation of cancer cells (e.g., gastric cancer cells, acute myeloid leukemia (AML) cells). In an exemplary assay, cancer cells (e.g., AGN, MKN45 gastric cancer cells) are cultured in the presence of IL-11, with or without an IL-11Rα binding protein. For AML cells, the cells may also be cultured in the presence of G-CSF. Cell proliferation is then measured using standard techniques, and / or by assessing the formation of L-CFU in the case of AML cells, for example, as discussed herein. Exemplary assays applicable to the present disclosure are included in Zhang et al., Int J Biol Sci., 8: 383-393, 2012 and Kimura et al., Leukemia, 13: 1018-1027, 1999.

[0117] For illustrative purposes only, the binding interaction between IL-11Rα and an antibody, or antigen-binding fragment thereof, or the binding / signaling between IL-11Rα and IL-11, as described herein, can be detected and quantified using a variety of routine methods, including Biacore® assays (e.g., coupled to a sensor chip using appropriately tagged soluble reagents), FACS analysis using cells (either natural or recombinant) expressing IL-11Rα on the cell surface, immunoassays, fluorescent staining assays, ELISA assays, and microcalorimetry approaches, such as ITC (isothermal titration calorimetry), etc. Similarly, the functional properties of anti-IL-11Rα antibodies can be evaluated using a variety of methods known to those skilled in the art, such as affinity / binding assays (e.g., surface plasmon resonance, competitive inhibition assays); cytotoxicity assays, cell survival assays, cell proliferation or differentiation assays, cancer cell and / or tumor growth inhibition using in vitro or in vivo models. Other assays may test the ability of the antibodies described herein to block normal IL-11Rα-mediated responses. The antibodies described herein may also be tested for in vitro and in vivo efficacy. Such assays may be performed using well-established protocols known to those skilled in the art (see, e.g., Current Protocols in Molecular Biology (Greene Publ. Assoc. Inc. & John Wiley & Sons, Inc., NY, NY); Current Protocols in Immunology (Edited by: John E. Coligan, Ada M. Kruisbeek, David H. Margulies, Ethan M. Shevach, Warren Strober 2001 John Wiley & Sons, NY, NY); or commercially available kits.

[0118] In certain embodiments, the Fc region of the antibody, or antigen-binding fragment thereof, comprises, consists of, or consists essentially of an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or a hybrid and / or variant thereof. In certain embodiments, the Fc region comprises, consists of, or consists essentially of an Fc from human IgG1 or IgG4 (see, e.g., Allberse and Schuurman, Immunology. 105:9-19, 2002), or a fragment or variant thereof. Table F1 below provides exemplary sequences (CH1, hinge (underlined), CH2, and CH3 regions) from human IgG4. Examples of variant IgG4 sequences that can be used include the S228P / S241P variant. [Table 4]

[0119] In certain embodiments, antibodies or antigen-binding fragments thereof comprise variant or otherwise modified Fc regions, including those with altered properties or biological activity compared to a wild-type Fc region. Examples of modified Fc regions include regions having a mutated sequence, e.g., by substitution, insertion, deletion, or truncation of one or more amino acids relative to the wild-type sequence, hybrid Fc polypeptides composed of domains from different immunoglobulin classes / subclasses, Fc polypeptides with altered glycosylation / sialylation patterns, and Fc polypeptides that are modified or derivatized, e.g., by biotinylation (see, e.g., U.S. Patent Application Publication No. 2010 / 0209424), phosphorylation, sulfation, the like, or any combination of the foregoing. Such modifications may affect the binding characteristics of the Fc region to one or more specific FcRs (e.g., FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, FcγRIIIb, FcRn), its pharmacokinetic properties (e.g., stability or half-life, bioavailability, tissue distribution, volume of distribution, concentration, elimination rate constant, elimination rate, area under the curve (AUC), clearance, Cmax , t max , C min , variation), its immunogenicity, its complement fixation or activation, and / or CDC / ADCC / ADCP-related activities of the Fc region relative to the corresponding wild-type Fc sequence of the antibody or antigen-binding fragment thereof, among other properties described herein. Included are modified Fc regions of human and / or murine origin.

[0120] In certain embodiments, the antibody or antigen-binding fragment thereof comprises a hybrid Fc region, e.g., a hybrid Fc region that comprises Fc domains (e.g., hinge, CH domain, and / or nucleotide sequences) from immunoglobulins of different species (e.g., human, mouse), different Ig classes, and / or different Ig subclasses. 2 , C.H. 3 , C.H. 4) combination. Also included are antibodies or antigen-binding fragments thereof that comprise derivatized or otherwise modified Fc regions. In certain aspects, the Fc region is modified, e.g., by phosphorylation, sulfation, acylation, glycosylation, methylation, farnesylation, acetylation, amidation, and the like, as compared to a wild-type or native Fc region. In certain embodiments, the Fc region comprises a wild-type or native glycosylation pattern, or alternatively, it comprises increased glycosylation as compared to the native form, decreased glycosylation as compared to the native form, or it is completely deglycosylated. As an example of a modified Fc glycoform, reduced glycosylation of the Fc region reduces binding to the C1q region of the first complement component C1, a decrease in ADCC-related activity, and / or a decrease in CDC-related activity. Certain embodiments thus employ a deglycosylated or non-glycosylated Fc region. For the production of an exemplary non-glycosylated Fc region, see, e.g., WO2005 / 047337. Another example of an Fc region glycoform is generated by substituting position Q295 with a cysteine ​​residue according to the Kabat et al. numbering system (see, e.g., U.S. Patent Application No. 2010 / 0080794). Certain embodiments include an Fc region, in which about 80-100% of the glycoproteins in the Fc region comprise a mature core carbohydrate structure lacking fucose (see, e.g., U.S. Patent Application No. 2010 / 0255013). Some embodiments include Fc regions that are optimized by substitutions or deletions to reduce the level of fucosylation, e.g., to increase affinity for FcγRI, FcγRIa, or FcγRIIIa, and / or to improve phagocytosis by FcγRIIa-expressing cells (see U.S. Patent Application Nos. 2010 / 0249382 and 2007 / 0148170).

[0121] As another example of a modified Fc glycoform, the Fc region of an antibody or antigen-binding fragment thereof may comprise oligomannose-type N-glycans, optionally having one or more of the following: increased ADCC effector activity, increased binding affinity for FcγRIIIA (and certain other FcRs), similar or increased binding specificity for a target of an IL-11Rα polypeptide, similar or higher binding affinity for a target of an IL-11Rα polypeptide, and / or similar or lower binding affinity for a mannose receptor, compared to a corresponding Fc region comprising complex-type N-glycans (see, e.g., U.S. Patent Application Publication No. 2007 / 0092521 and U.S. Patent No. 7,700,321). As another example, enhanced affinity of an Fc region for FcγR has been achieved using engineered glycoforms generated by expression of the antibody in engineered or variant cell lines (see, e.g., Umana et al., Nat Biotechnol. 17:176-180, 1999; Davies et al., Biotechnol Bioeng. 74:288-294, 2001; Shields et al., J Biol Chem. 277:26733-26740, 2002; Shinkawa et al., 2003, J Biol Chem. 278:3466-3473, 2003; and U.S. Patent Application No. 2007 / 0111281). Certain Fc region glycoforms contain an increased proportion of N-glycosidically linked complex glycans that do not have the 1-position of fucose attached to the 6-position of the N-acetylglucosamine at the reducing end of the glycan (see, e.g., U.S. Patent Application No. 2010 / 0092997). Certain embodiments may include an IgG Fc region that is glycosylated with at least one galactose moiety linked to each terminal sialic acid moiety by an α-2,6 linkage, optionally wherein the Fc region has greater anti-inflammatory activity compared to a corresponding, wild-type Fc region (see, e.g., U.S. Patent Application No. 2008 / 0206246).Some of these and related altered glycosylation approaches, as described herein, have produced substantial enhancements in the ability of the Fc region to selectively bind to FcRs, such as FcγRIII, mediate ADCC, and modify other properties of the Fc region.

[0122] Particular variants, fragments, hybrids, or otherwise modified Fc regions of antibodies or antigen-binding fragments thereof may have altered binding to one or more FcRs and / or corresponding changes in effector function compared to the corresponding wild-type Fc sequence (e.g., same species, same Ig class, same Ig subclass). For example, such Fc regions may have increased binding to one or more of Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In other embodiments, variants, fragments, hybrids, or modified Fc regions may have decreased binding to one or more of Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. Particular FcRs are described elsewhere herein.

[0123] In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to increase binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an IgG1 or IgG3 Fc domain comprising one or more mutations to increase binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to increase effector function. In some embodiments, at least one antibody comprises an Fc domain selected from human IgG1 and IgG3 comprising one or more mutations to increase effector function.

[0124] In some embodiments, the antibody is a blocking antibody comprising an Fc domain with high effector activity. In some embodiments, the blocking antibody comprises an Fc domain selected from human IgG1 and IgG3 comprising one or more mutations to increase effector function. In some embodiments, the antibody is a partial blocking antibody comprising an Fc domain with high effector activity. In some embodiments, the partial blocking antibody comprises an Fc domain selected from human IgG1 and IgG3 comprising one or more mutations to increase effector function. In some embodiments, the antibody is a non-blocking antibody comprising an Fc domain with high effector activity. In some embodiments, the non-blocking antibody comprises an Fc domain selected from human IgG1 or IgG3 comprising one or more mutations to increase effector function.

[0125] In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to reduce binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an IgG1 or IgG3 Fc domain comprising one or more mutations to reduce binding to one or more Fcγ receptors, Fcα receptors, Fcε receptors, and / or neonatal Fc receptors compared to the corresponding wild-type Fc sequence. In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to reduce effector function. In some embodiments, the antibody comprises an Fc domain selected from human IgG2 and IgG4 comprising one or more mutations to reduce effector function.

[0126] In some embodiments, the antibody is a blocking antibody comprising an Fc domain with reduced effector activity. In some embodiments, the blocking antibody comprises an Fc domain selected from human IgG2 and IgG4 comprising one or more mutations to reduce effector function. In some embodiments, the antibody is a partial blocking antibody comprising an Fc domain with reduced effector activity. In some embodiments, the partial blocking antibody comprises an Fc domain selected from human IgG2 and IgG4 comprising one or more mutations to reduce effector function. In some embodiments, the antibody is a non-blocking antibody comprising an Fc domain with reduced effector activity. In some embodiments, the non-blocking antibody comprises an Fc domain selected from human IgG2 and IgG4 comprising one or more mutations to reduce effector function.

[0127] Specific examples of Fc variants with altered (e.g., increased, decreased) effector function / FcR binding are described, for example, in U.S. Patent Nos. 5,624,821 and 7,425,619; U.S. Patent Application Nos. 2009 / 0017023, 2009 / 0010921, and 2010 / 0203046; and WO2000 / 42072 and WO2004 / 016750. Particular examples include human Fc regions with one or more substitutions at positions 298, 333, and / or 334, e.g., S298A, E333A, and / or K334A (based on the EU index numbering of Kabat et al.), which have been shown to increase binding to the activating receptor FcγRIIIa and decrease binding to the inhibitory receptor FcγRIIb. These mutations can be combined to obtain double and triple mutant variants with further improvements in binding to FcR. Particular embodiments include the S298A / E333A / K334A triple mutant, which has increased binding to FcγRIIIa, decreased binding to FcγRIIb, and increased ADCC (see, e.g., Shields et al., J Biol Chem. 276:6591-6604, 2001; and Presta et al., Biochem Soc Trans. 30:487-490, 2002). See also Umana et al. (supra); and engineered Fc glycoforms with increased binding to FcR as disclosed in U.S. Pat. No. 7,662,925. Some embodiments include an Fc region that includes one or more substitutions selected from 434S, 252Y / 428L, 252Y / 434S, and 428L / 434S, based on the EU index of Kabat et al. (see U.S. Patent Application Publication Nos. 2009 / 0163699 and 20060173170).

[0128] Certain variant, fragment, hybrid, or modified Fc regions may have altered effector functions compared to the corresponding wild-type Fc sequence. For example, such Fc regions may have increased complement fixation or activation, increased Clq binding affinity, increased CDC-related activity, increased ADCC-related activity, and / or increased ADCP-related activity compared to the corresponding wild-type Fc sequence. In other embodiments, such Fc regions may have decreased complement fixation or activation, decreased Clq binding affinity, decreased CDC-related activity, decreased ADCC-related activity, and / or decreased ADCP-related activity compared to the corresponding wild-type Fc sequence. As merely one illustrative example, the Fc region may include deletions or substitutions in complement binding sites, such as C1q binding sites, and / or deletions or substitutions in ADCC sites. Examples of such deletions / substitutions are described, for example, in U.S. Patent No. 7,030,226. Many Fc effector functions, such as ADCC, can be assayed according to routine techniques in the art (see, e.g., Zuckerman et al., CRC Crit Rev Microbiol. 7:1-26, 1978). Useful effector cells for such assays include, but are not limited to, natural killer (NK) cells, macrophages, and other peripheral blood mononuclear cells (PBMCs). Alternatively, or additionally, certain Fc effector functions can be assessed in vivo, for example, by using animal models described in Clynes et al. PNAS. 95:652-656, 1998.

[0129] Certain variant hybrids or modified Fc regions may have altered stability or half-life compared to the corresponding wild-type Fc sequence. In certain embodiments, such Fc regions may have increased half-life compared to the corresponding wild-type Fc sequence. In other embodiments, variant hybrids or modified Fc regions may have decreased half-life compared to the corresponding wild-type Fc sequence. Half-life can be measured in vitro (e.g., under physiological conditions) or in vivo according to routine techniques in the art, such as radiolabeling, ELISA, or other methods. In vivo measurements of stability or half-life can be measured in one or more body fluids, including blood, serum, plasma, urine, or cerebrospinal fluid, or in a given tissue, such as liver, kidney, muscle, central nervous system tissue, bone, etc. As an example, modifications to an Fc region that alter its ability to bind to FcRn can alter its half-life in vivo. Non-limiting examples of assays for measuring in vivo pharmacokinetic properties (e.g., in vivo mean elimination half-life) and Fc modifications that alter their binding to FcRn are described, for example, in U.S. Pat. Nos. 7,217,797 and 7,732,570; and U.S. Patent Application Nos. US2010 / 0143254 and 2010 / 0143254.

[0130] Additional non-limiting examples of stability or half-life altering modifications include the CH 2 251-256, 285-290, and 308-314 in the domain, as well as CH 3The substitution / deletion of one or more of the amino acid residues selected from 385-389 and 428-436 in the domain is included. See US Patent Application Publication No. 2003 / 0190311. Specific examples include substitution with leucine at position 251, substitution with tyrosine, tryptophan or phenylalanine at position 252, substitution with threonine or serine at position 254, substitution with arginine at position 255, substitution with glutamine, arginine, serine, threonine or glutamic acid at position 256, substitution with threonine at position 308, substitution with proline at position 309, substitution with serine at position 311, substitution with aspartic acid at position 312, substitution with leucine at position 314, substitution with arginine, aspartic acid at position 316, substitution with leucine at position 385, substitution with arginine, aspartic acid at position 318, substitution with leucine at position 386, substitution with leucine at position 387, substitution with leucine at position 389, substitution with leucine at position 388, substitution with leucine at position 389 ... substitution with arginine or serine, substitution with threonine or proline at position 386, substitution with arginine or proline at position 387, substitution with proline, asparagine or serine at position 389, substitution with methionine or threonine at position 428, substitution with tyrosine or phenylalanine at position 434, substitution with histidine, arginine, lysine or serine at position 433, and / or substitution with histidine, tyrosine, arginine or threonine at position 436, including any combination thereof. Such modifications optionally increase the affinity of the Fc region for FcRn, thereby increasing the half-life compared to the corresponding, wild-type Fc region.

[0131] Certain variant hybrids, or modified Fc regions, may have altered solubility compared to the corresponding wild-type Fc sequence. In certain embodiments, such Fc regions may have increased solubility compared to the corresponding wild-type Fc sequence. In other embodiments, variant hybrids, or modified Fc regions may have decreased solubility compared to the corresponding wild-type Fc sequence. Solubility can be measured, for example, in vitro (e.g., under physiological conditions) according to routine techniques in the art. Exemplary solubility measurements are described elsewhere herein.

[0132] A variant Fc region can also have one or more mutated hinge regions, e.g., as described in US Patent Application Publication No. 2003 / 0118592. For example, one or more cysteines in the hinge region can be deleted or replaced with different amino acids. The mutated hinge region can contain no cysteine ​​residues, or it can contain one, two, or three fewer cysteine ​​residues than the corresponding wild-type hinge region. In some embodiments, Fc regions with this type of mutated hinge region exhibit reduced dimerization ability compared to the wild-type Ig hinge region.

[0133] In certain embodiments, the antibody or antigen-binding fragment thereof is capable of being incubated for about or at least about 30 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 12 hours, about 18 hours, about 20 hours, about 24 hours at about pH 7.4, at about physiological pH, at about 25° C. or room temperature, and / or at about 37° C. or human body temperature (e.g., in vivo, in serum, in a given tissue, in a given species, such as rat, mouse, monkey, or human). or about 30 hours, about 36 hours, about 40 hours, about 48 hours, about 50 hours, about 60 hours, about 70 hours, about 72 hours, about 80 hours, about 84 hours, about 90 hours, about 96 hours, about 120 hours, or about 144 hours or more, or about 1 week, or about 2 weeks, or about 3 weeks, or about 4 weeks, or about 5 weeks, or about 6 weeks or more, or any intervening half-life, including all ranges therebetween.

[0134] In some embodiments, the antibody or antigen-binding fragment thereof has a T of about or at least about 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or 75° C. m In some embodiments, the antibody or antigen-binding fragment thereof has a T of about 65° C. or higher, e.g., in PBS (phosphate buffered saline). m has.

[0135] In some embodiments, the antibody or antigen-binding fragment thereof is conjugated to one or more cytotoxic or chemotherapeutic agents. General examples of cytotoxic or chemotherapeutic agents include, but are not limited to, alkylating agents, antimetabolites, anthracyclines, antitumor antibiotics, platinums, type I topoisomerase inhibitors, type II topoisomerase inhibitors, vinca alkaloids, and taxanes. Specific examples of cytotoxic or chemotherapeutic agents include, but are not limited to, cyclophosphamide, cilengitide, lomustine (CCNU), melphalan, procarbazine, carmustine (BCNU), enzastaurin, busulfan, daunorubicin, doxorubicin, gefitinib, erlotinib. Idarubicin, temozolomide, epirubicin, mitoxantrone, bleomycin, cisplatin, carboplatin, oxaliplatin, camptothecin, irinotecan, topotecan, amsacrine, etoposide, etoposide phosphate, teniposide, temsirolimus, everolimus, vincristine, vinblastine, vinorelbine, vindesine, CT52923, paclitaxel, imatinib, dasatinib, sorafenib, pazopanib, sunitonib, vatalanib, gefitinib, These include futinib, erlotinib, AEE-788, dichloroacetic acid, tamoxifen, fasudil, SB-681323, semaxanib, donepizil, galantamine, memantine, rivastigmine, tacrine, rasigline, naltrexone, lubiprostone, safinamide, istradefylline, pimavanserin, pitolisant, isradipine, pridopidine (ACR16), tetrabenazine, bexarotene, glatirimer acetate, fingolimod, and mitoxantrone, including pharmaceutical acceptable salts and acids thereof. Further examples of cytotoxic or chemotherapeutic agents include alkylating agents such as thiotepa, cyclophosphamide (CYTOXAN™), and the like; alkylsulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, uredopa, and the like; ethylenimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine;Nitrogen mustards, such as chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobembitine, phenesterine, prednimustine, trofosfamide, uracil mustard, etc.; nitrosureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine, etc.; antibiotics, such as aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, ca Richeamicin, Carabicin, Carminomycin, Carzinophilin, Chromomycin, Dactinomycin, Daunorubicin, Detorubicin, 6-Diazo-5-Oxo-L-Norleucine, Doxorubicin, Epirubicin, Esorubicin, Idarubicin, Martilomycin, Mitomycin, Mycophenolic Acid, Nogalamycin, Olivomycin, Peplomycin, Potofilomycin, Puromycin, Querramycin, Rodorubicin, Streptonigrin, Streptozocin, Tubercidin, Ubenimex, Zinostatin, Zorubicin, etc.; Metabolism antagonists, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues, such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues, such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU; androgens, such as calsterone, dromostanolone propionate, enocitabine, 5-FU, etc. Pitiostanol, mepitiostane, testolactone, etc.; antiadrenal drugs, such as aminoglutethimide, mitotane, trilostane, etc.; folic acid supplements, such as furoic acid; aceglatone; aldophosphamide glycosides; aminolevulinic acid; amsacrine; bestravcil; bisantrene; edatraxate; deformamine; demecolcine; diaziquone; erformitin; elliptinium acetate; etoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin;Phenamet; pirarubicin; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK; razoxane; sizofiran; spirogermanium; tenuazonic acid; triazicon; 2,2',2"-trichlorotriethylamine; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, such as paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, NJ) and doxetaxel (TAXOTERE®, Rhone-Poulenc Rolles, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogues such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitors RFS 2000; difluoromethylorutin (DMFO); retinoic acid derivatives such as Targretin™ (bexarotene); Panretin™ (alitretinoin); ONTAK™ (denileukin diftitox); esperamicin; capecitabine; and pharma- ceutical acceptable salts, acids, or derivatives of any of the above.

[0136] In some embodiments, the antibodies disclosed herein are conjugated or operably linked to a radioisotope to form radioconjugates and / or macrocyclic chelators useful for complexing radioactive metal ions. A variety of radioisotopes are available for the production of radioconjugated antibodies. Examples include, but are not limited to: 90 Y, 123 I, 125 I, 131 I, 186 Re, 188 Re, 211 At, and 212In certain embodiments, the macrocyclic chelator is 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), which can be attached to the antibody via a linker molecule. Such linker molecules are generally known in the art and have been described in Denardo et al., 1998, Clin Cancer Res. 4:2483-90; Peterson et al., 1999, Bioconjug. Chem. 10:553; and Zimmerman et al., 1999, Nucl. Med. Biol. 26:943-50.

[0137] Other modifications of the antibodies (and polypeptides) of the disclosure are also contemplated herein. For example, in some embodiments, the antibodies are linked to one of a variety of nonproteinaceous polymers, such as polyethylene glycol, polypropylene glycol, polyoxyalkylenes, or copolymers of polyethylene glycol and polypropylene glycol. In some embodiments, the antibodies are encapsulated in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin-microcapsules and poly-(methyl methacylate) microcapsules, respectively), in colloid drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16th edition, Oslo, A., Ed., (1980).

[0138] The antibodies or antigen-binding fragments thereof may be used in any of the compositions, methods, and / or kits described herein and may be combined with one or more of the additional agents described herein.

[0139] Methods of Use and Pharmaceutical Compositions Certain embodiments relate to a method of treating, ameliorating, and / or slowing the progression of a disease or condition in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding fragment thereof that binds to interleukin-11 receptor subunit alpha (IL-11Rα) as described herein, or a pharmaceutical composition comprising the same. In some examples, the antibody or antigen-binding fragment thereof antagonizes the binding / signaling activity between IL-11Rα and its ligand IL-11. In some embodiments, the disease or condition is an IL-11-related or IL-11-mediated disease or condition. In some embodiments, the disease or condition is cancer, an inflammatory disease, an autoimmune disease, a wasting disease, a bone disease, or a fibrotic disease.

[0140] In some embodiments, the disease or condition is a cancer or tumor, as described above. For example, in some cases, the cancer expresses or overexpresses IL-11Rα and / or IL-11, and in some cases, the cancer exhibits IL-11Rα / IL-11-dependent growth, adhesion, migration, invasion, and / or chemotherapy resistance. In some cases, the cancer is a primary cancer. In some cases, the cancer is a metastatic cancer.

[0141] Exemplary cancers include, but are not limited to, bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma, lymphoma), hepatocellular carcinoma (liver cell carcinoma), sarcoma, B-cell malignancies, breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, uterine cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer. In certain embodiments, the cancer is a metastatic cancer, e.g., that has metastasized to bone.

[0142] In some embodiments, the methods and compositions described herein increase the median survival of a subject by 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 15 weeks, 20 weeks, 25 weeks, 30 weeks, 40 weeks, or more. In certain embodiments, the methods and compositions described herein increase the median survival of a subject by 1 year, 2 years, 3 years, or more. In some embodiments, the methods and compositions described herein increase progression-free survival by 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, or more. In certain embodiments, the methods and compositions described herein increase progression-free survival by 1 year, 2 years, 3 years, or more.

[0143] In certain embodiments, the methods and compositions described herein are sufficient to cause a statistically significant reduction in the amount of viable tumor, e.g., at least a 10%, 20%, 30%, 40%, 50% or greater reduction in tumor mass, or tumor regression as indicated by altered (e.g., decreased with statistical significance) scan dimensions. In some embodiments, the methods and compositions described herein reduce the rate of cancer growth (e.g., in vivo or in vitro, including cancer cells isolated from a biopsy or other sample and grown in vitro) by about or at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000% or more compared to an untreated control. In some examples, the methods and compositions described herein reduce cancer cell initiation, migration, adhesion, invasiveness, and / or metastasis by about or at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000% or more compared to an untreated control. In some examples, the methods and compositions described herein reduce angiogenesis in a tumor environment by about or at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000% or more compared to an untreated control.

[0144] In certain embodiments, the disease or condition is an inflammatory disease. Non-limiting examples of inflammatory diseases and conditions include airway or lung inflammation (e.g., inflammatory lung disease), asthma, rhinitis, chronic obstructive pulmonary disorder (COPD), dermatitis, psoriasis, hepatitis, gastritis, irritable bowel syndrome (IBS), ulcerative colitis, Crohn's disease, colitis, diverticulitis, lupus erythematosus, nephritis, Parkinson's disease, multiple sclerosis (MS), Alzheimer's disease, arthritis, rheumatoid arthritis, sepsis, infection-induced inflammation, cardiovascular disease, such as atherosclerosis and vasculitis, diabetes, and gout.

[0145] In certain embodiments, the disease or condition is an autoimmune disease. Non-limiting examples of autoimmune diseases and conditions include arthritis (including rheumatoid arthritis, reactive arthritis), systemic lupus erythematosus (SLE), psoriasis, inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, encephalomyelitis, uveitis, myasthenia gravis, multiple sclerosis, insulin-dependent diabetes mellitus, Addison's disease, celiac disease, chronic fatigue syndrome, autoimmune hepatitis, autoimmune alopecia, ankylosing spondylitis, fibromyalgia, pemphigus vulgaris, Sjogren's syndrome, Kawasaki disease, hyperthyroidism / Graves' disease, hypothyroidism / Hashimoto's disease, endometriosis, scleroderma, pernicious poverty. Autoimmune hemolytic anemia, including idiopathic thrombocytopenic purpura, idiopathic hemolytic anemia, pulmonary embolism, pulmonary edema, pulmonary edema, pulmonary malformation, pulmonary malformation, pulmonary pulmonary fibrosis, ...

[0146] In certain embodiments, the disease or condition is a debilitating disease. Non-limiting examples of debilitating diseases and conditions include cachexia, including cachexia associated with cancer or renal failure, and sarcopenia. In certain embodiments, the disease or condition is a bone disease. Non-limiting examples of bone diseases and conditions include osteoporosis (including postmenopausal osteoporosis), bone fractures, Paget's disease of bone, and bone resorption / damage associated with cancer or cancer treatment, including chemotherapy, hormone ablation, and hormone inhibition.

[0147] In some embodiments, the disease or condition is fibrosis or a fibrotic disease. Examples include fibrosis of the lung, cardiovascular system, liver, brain, joints (e.g., knee, hip, ankle, ankle, shoulder, elbow, wrist, hand, spine), gut, skin, kidney, liver, thyroid, bone marrow, retroperitoneal, and eye (see, e.g., Schafer et al., Nature. 552: 110-115, 2017; and Ng et al., Sci Transl Med. 2019 Sep 25; 11(511)).

[0148] In some embodiments, the pulmonary fibrosis is selected from fibrothorax, pulmonary fibrosis (e.g., cystic fibrosis, interstitial lung disease (ILD), an autosomal recessive genetic disorder, such as Hermansky-Pudlak syndrome type 1, 2, 3, 4, 5, 6, 7, or 8, and radiation-induced lung injury. In some embodiments, the pulmonary fibrosis is associated with an ILD, such as idiopathic or secondary ILD. Examples of idiopathic ILD include idiopathic pulmonary fibrosis (IPF), desquamative interstitial pneumonia (DIP), acute interstitial pneumonia (AIP), also known as Hamman-Rich syndrome, nonspecific interstitial pneumonia (NSIP), respiratory bronchiolitis-associated interstitial lung disease (RB-ILD), idiopathic organizing pneumonia (COP), and lymphocytic interstitial pneumonia (LIP). Common examples of secondary ILD include ILD associated with connective tissue and autoimmune diseases (e.g., sarcoidosis, rheumatoid arthritis, systemic lupus erythematosus, systemic sclerosis, polymyositis, dermatomyositis, antisynthetase syndrome), inhalants (e.g., silicosis, asbestosis, beryllium disease, industrial printing chemicals, chronic hypersensitivity pneumonitis), drugs (drug-induced ILD, e.g., antibiotics, chemotherapeutic agents, antiarrhythmic agents), infections (e.g., SARS CoV-2, atypical pneumonia, Pneumocystis pneumonia, tuberculosis, Chlamydia trachomatis, respiratory syncytial virus), malignancies (lymphangitis carcinomatosa), and pediatric ILD, e.g., diffuse developmental disorders, growth abnormalities, alveolar hypoplasia, infantile disease of unknown etiology, and alveolar surfactant area-associated ILD.

[0149] In certain embodiments, the fibrosis of the cardiovascular system is myocardial fibrosis (eg, interstitial fibrosis, replacement fibrosis).

[0150] In some embodiments, the antibodies described herein are selective for both mouse and human IL-11Rα, blocking IL-11 signaling through both the STAT3 and ERK pathways, and abrogating both cis- and trans-IL-11 signaling. Following TGFβ stimulation, certain anti-huIL-11Rα antibodies described herein have been shown to reduce collagen expression in fibroblasts from primary IPF patients and reduce expression of procollagen I and TIMPs by PCLS from healthy donors. A similar reduction in procollagen release was observed in response to anti-huIL-11Rα in a preliminary PCLS study from IPF patients. In a mouse bleomycin pulmonary fibrosis study, blocking IL-11Rα signaling reduced pulmonary fibrosis and reduced BAL inflammatory cells when administered prophylactically or therapeutically after the onset of pulmonary fibrosis.

[0151] In certain embodiments, the methods and compositions described herein are sufficient to result in stable disease, hi certain embodiments, the methods and compositions described herein are sufficient to result in a clinically relevant reduction in symptoms of a particular disease indication known to a skilled clinician.

[0152] For in vivo use, certain embodiments include pharmaceutical compositions comprising the antibodies or antigen-binding fragments thereof described herein and a pharma- ceutically acceptable carrier. To prepare a therapeutic or pharmaceutical composition, an effective or desired amount of one or more agents is mixed with any pharmaceutical carrier or excipient known to those of skill in the art to be appropriate for the particular agent and / or mode of administration. The pharmaceutical carrier may be liquid, semi-liquid, or solid. Solutions or suspensions used for parenteral, intradermal, intraocular, subcutaneous, direct instillation into the bladder, or topical application may contain, for example, a sterile diluent (such as water), saline (e.g., phosphate buffered saline; PBS), fixed oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antimicrobial agents (such as benzyl alcohol and methylparabens); antioxidants (such as ascorbic acid and sodium bisulfite) and chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); buffers (such as acetates, citrates, and phosphates). If administered intravenously (e.g., by IV infusion), suitable carriers include saline or phosphate buffered saline (PBS), as well as solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, polypropylene glycol, and mixtures thereof.

[0153] Administration of the agents described herein in pure form or in suitable therapeutic or pharmaceutical compositions can be via any of the accepted modes of administration of agents to perform similar utilities. Therapeutic or pharmaceutical compositions can be prepared by combining the agent-containing composition with a suitable physiologically acceptable carrier, diluent, or excipient, and can be formulated into preparations in solid, semi-solid, liquid, or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols. In addition, other pharmacologic active ingredients (including other small molecules described elsewhere herein) and / or suitable excipients, such as salts, buffers, and stabilizers, may, but need not, be present in the composition.

[0154] Administration can be accomplished by a variety of different routes, including oral, parenteral, nasal, intravenous, intraocular, intradermal, intramuscular, subcutaneous, placement in the bladder, or topical. The preferred mode of administration will depend on the nature of the condition being treated or prevented. Certain embodiments include administration by IV infusion.

[0155] Carriers can include, for example, pharma- ceutical or physiologically acceptable carriers, excipients, or stabilizers that are non-toxic to cells or mammals exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carrier is an aqueous pH buffered solution. Examples of physiologically acceptable carriers include buffers, such as phosphates, citrates, other organic acids, and the like; antioxidants, including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, arginine, histidine, and / or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents, such as EDTA; sugar alcohols, such as mannitol or sorbitol; salt-forming counterions, such as sodium; and / or non-ionic surfactants, such as polysorbate 20 (TWEEN™), polyethylene glycol (PEG), poloxamer (PLURONICS™), and the like.

[0156] In some embodiments, one or more agents can be encapsulated in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin-microcapsules and poly-(methyl methacylate) microcapsules, respectively), in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules), or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16th edition, Oslo, A., Ed., (1980). The particles or liposomes may further comprise other therapeutic or diagnostic agents.

[0157] The exact dosage and duration of treatment is a function of the disease being treated and can be determined empirically using known test protocols or by testing the composition in model systems known in the art and extrapolating therefrom. Controlled clinical trials can also be performed. Dosage can also vary depending on the severity of the condition to be alleviated. Pharmaceutical compositions are generally formulated and administered to exert a therapeutically useful effect while minimizing undesirable side effects. The composition can be administered once or divided into multiple smaller doses administered at regular intervals. For any particular subject, the specific dosage regimen can be adjusted over time according to individual needs.

[0158] Typical routes of administration of these and related therapeutic or pharmaceutical compositions thus include, without limitation, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, ocular, rectal, vaginal, and intranasal. The term "parenteral" as used herein includes subcutaneous injection, intravenous, injection into the bladder, intramuscular, intrasternal injection, or infusion techniques. The therapeutic or pharmaceutical compositions according to certain embodiments of the present disclosure are formulated so that the active ingredients contained therein are bioavailable upon administration of the composition to a subject or patient. The composition administered to a subject or patient may take the form of one or more dosage units, where, for example, a tablet may be a single dosage unit and a container of the medicament described herein in aerosol form may hold multiple dosage units. Actual methods of preparing such dosage forms are known or will be apparent to those skilled in the art; see, for example, Remington: The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000). The composition to be administered typically contains a therapeutically effective amount of an agent described herein for treatment of the disease or condition of interest.

[0159] The therapeutic or pharmaceutical composition can be in solid or liquid form. In some embodiments, the carrier is particulate, and the composition is, for example, in tablet or powder form. The carrier can be liquid, and the composition is, for example, an oral oil, an injectable liquid, or an aerosol, which is useful, for example, in inhalation administration. When intended for oral administration, the pharmaceutical composition is preferably in either solid or liquid form, where semi-solid, semi-liquid, suspension, and gel forms are included within the forms considered herein as either solid or liquid. Particular embodiments include sterile injectable solutions.

[0160] As a solid composition for oral administration, the pharmaceutical composition may be formulated into powder, granules, gels, compressed tablets, pills, capsules, chewing gum, wafers or the like. Such solid compositions typically contain one or more inert diluents or edible carriers. Also, one or more of the following may be present: binders, such as carboxymethylcellulose, ethylcellulose, microcrystalline cellulose, tragacanth or gelatin; excipients, such as starch, lactose or dextrin; disintegrants, such as alginic acid, sodium alginate, primogel, corn starch and the like; lubricants, such as magnesium stearate or stereotex; glidants, such as colloidal silicon dioxide; sweeteners, such as sucrose or saccharin; flavorings, such as peppermint, methyl salicylate or orange flavoring; and coloring agents. When the pharmaceutical composition is in the form of a capsule, such as a gelatin capsule, it may contain, in addition to the above types of materials, liquid carriers, such as polyethylene glycol or oil.

[0161] The therapeutic or pharmaceutical composition may be in the form of a liquid, such as an elixir, syrup, solution, gel, emulsion, or suspension. The liquid may be for oral administration or for delivery by injection, as two examples. When intended for oral administration, a preferred composition contains, in addition to the compound, one or more of a sweetener, a preservative, a dye / colorant, and a flavor enhancer. In a composition intended to be administered by injection, one or more of a surfactant, a preservative, a wetting agent, a dispersing agent, a suspending agent, a buffer, a stabilizer, and an isotonic agent may be included.

[0162] Liquid therapeutic or pharmaceutical compositions, whether they are in solution, suspension, or other similar form, may contain one or more of the following adjuvants: sterile diluents, such as water for injection, saline, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils, such as synthetic mono- or diglycerides, polyethylene glycols, glycerin, propylene glycol, or other solvents that serve as solvents or suspending media; antibacterial agents, such as benzyl alcohol or methylparabens; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates, phosphates, and agents for adjusting tonicity, such as sodium chloride or glucose. Parenteral preparations can be enclosed in ampoules, disposable syringes, or multiple dose vials made of glass or plastic. Physiological saline is the preferred adjuvant. Pharmaceutical compositions for injection are preferably sterile.

[0163] Liquid therapeutic or pharmaceutical compositions intended for either parenteral, ocular, or oral administration should contain an amount of drug such that a suitable dosage is obtained. Typically, this amount is at least 0.01% of the drug of interest in the composition. When intended for oral administration, this amount can vary to be between 0.1 and about 70% by weight of the composition. Certain oral therapeutic or pharmaceutical compositions contain between about 4% and about 75% of the drug of interest. In certain embodiments, therapeutic or pharmaceutical compositions and preparations are prepared such that a parenteral dosage unit contains between 0.01 and 10% by weight of the drug of interest before dilution.

[0164] The therapeutic or pharmaceutical composition may be intended for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment, or gel base. The base may, for example, comprise one or more of the following: petrolatum, lanolin, polyethylene glycol, beeswax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Viscosifiers may be present in therapeutic or pharmaceutical compositions for topical administration. If intended for transdermal administration, the composition may comprise a transdermal patch or iontophoresis device.

[0165] Therapeutic or pharmaceutical compositions may be intended for rectal administration, for example, in the form of a suppository that dissolves in the rectum and releases the drug. Compositions for rectal administration may contain an oleaginous base as a suitable non-irritating excipient. Such bases include, but are not limited to, lanolin, cocoa butter, and polyethylene glycol.

[0166] Therapeutic or pharmaceutical compositions may contain various materials that modify the physical form of a solid or liquid dosage unit. For example, the composition may contain a material that forms a coating shell around the active ingredient. The material that forms the coating shell is typically inert and may be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredient may be encapsulated in a gelatin capsule. Therapeutic or pharmaceutical compositions in solid or liquid form may contain a component that binds to the drug and thereby assists in the delivery of the compound. Suitable components that may act in this capacity include monoclonal or polyclonal antibodies, one or more proteins, or liposomes.

[0167] The therapeutic or pharmaceutical composition may consist essentially of a dosage unit that can be administered as an aerosol. The term "aerosol" is used to denote a variety of systems ranging from those of colloidal nature to systems consisting of pressurized packages. Delivery may be by liquefied or compressed gas or by a suitable pump system that dispenses the active ingredient. The aerosol may be delivered in a single phase, two phase, or three phase system to deliver the active ingredient. Delivery of the aerosol includes the necessary containers, activators, valves, subcontainers, and the like, which may together form a kit. Those skilled in the art may determine the preferred aerosol without undue experimentation.

[0168] The compositions described herein may be prepared with carriers that protect the agents against rapid elimination from the body, such as time release formulations or coatings. Such carriers include controlled release formulations, including, but not limited to, implants and microencapsulated delivery systems, and biodegradable, biocompatible polymers, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, polyorthoesters, polylactic acid, and others known to those skilled in the art.

[0169] Pharmaceutical compositions can be prepared by methodologies well known in the pharmaceutical arts. For example, therapeutic or pharmaceutical compositions intended to be administered by injection may be accompanied by sterile distilled water to form a solution and may contain one or more salts, buffers and / or stabilizers. Surfactants may be added to facilitate the formation of a homogeneous solution or suspension. Surfactants are compounds that non-covalently interact with drugs to facilitate the dissolution or homogeneous suspension of the drug in an aqueous delivery system.

[0170] Therapeutic or pharmaceutical compositions may be administered in a therapeutically effective amount, which will vary depending on a variety of factors, including the activity of the particular compound used; the metabolic stability and time of action of the compound; the age, weight, general health, sex, and diet of the subject; the mode and time of administration; the excretion rate; the drug combination; the severity of the particular disorder or condition; and the subject being treated. In some cases, a therapeutically effective daily dose is from about 0.001 mg / kg (i.e., 0.07 mg) to about 100 mg / kg (i.e., 7.0 g) (for a mammal weighing 70 kg); preferably, a therapeutically effective dose is from about 0.01 mg / kg (i.e., 0.7 mg) to about 50 mg / kg (i.e., 3.5 g) (for a mammal weighing 70 kg); more preferably, a therapeutically effective amount is from about 1 mg / kg (i.e., 70 mg) to about 25 mg / kg (i.e., 1.75 g) (for a mammal weighing 70 kg). In some embodiments, the therapeutically effective dose is administered weekly, biweekly, or monthly. In certain embodiments, the therapeutically effective dose is administered weekly, biweekly, or monthly, e.g., at a dose of about 1-10 or 1-5 mg / kg, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg.

[0171] Also included are patient care kits comprising (a) an antibody or antigen-binding fragment thereof that binds IL-11Rα, as described herein; and, optionally, (b) at least one additional therapeutic agent. In certain kits, (a) and (b) are in separate therapeutic compositions. In some kits, (a) and (b) are in the same therapeutic composition.

[0172] The kits herein may also include one or more additional therapeutic agents or other components appropriate or desirable for the indication being treated or for the desired diagnostic application. The kits herein may also include one or more syringes or other components necessary or desirable to facilitate the intended mode of delivery (e.g., stents, implantable depots, etc.).

[0173] In some embodiments, the patient care kit includes separate containers, dividers, or compartments for the composition and the informational material. For example, the composition may be included in a bottle, vial, or syringe, and the informational material can be included in association with the container. In some embodiments, the separate elements of the kit are included within a single, undivided container. For example, the composition is included in a bottle, vial, or syringe that has informational material in the form of a label attached thereto. In some embodiments, the kit includes multiple (e.g., packs) of individual containers, each including one or more unit dosage forms (e.g., dosage forms described herein) of the antibody and, optionally, at least one additional therapeutic agent. For example, the kit includes multiple syringes, ampoules, foil packets, or blister packs, each including a single unit dose of the antibody and, optionally, at least one additional therapeutic agent. The containers of the kit can be airtight, waterproof (e.g., impermeable to changes in moisture or evaporation), and / or light-tight.

[0174] The patient care kit optionally includes a device suitable for administration of the composition, such as a syringe, inhaler, dropper (e.g., eye dropper), swab (e.g., cotton swab or wooden swab), or any such delivery device. In some embodiments, the device is an implantable device that dispenses a metered dose of the agent. Also included are methods of providing the kit, such as by combining the components described herein.

[0175] Expression and purification system Certain embodiments include methods and related compositions for expressing and purifying the anti-IL-11Rα antibodies or antigen-binding fragments thereof described herein. Such recombinant anti-IL-11Rα antibodies can be conveniently prepared using standard protocols described, for example, in Sambrook, et al., (1989, supra), especially sections 16 and 17; Ausubel et al., (1994, supra), especially chapters 10 and 16; and Coligan et al., Current Protocols in Protein Science (John Wiley & Sons, Inc. 1995-1997), especially chapters 1, 5, and 6. As one general example, an anti-IL-11Rα antibody can be prepared by a procedure including one or more of the following steps: (a) preparing a construct comprising a polynucleotide sequence encoding an anti-IL-11Rα antibody heavy and / or light chain and operably linked to regulatory elements; (b) introducing the construct into a host cell; (c) culturing the host cell to express the anti-IL-11Rα antibody; and (d) isolating the anti-IL-11Rα from the host cell.

[0176] Particular embodiments thus include polynucleotides encoding the anti-IL-11Rα antibodies or antigen-binding fragments thereof described herein, vectors comprising said polynucleotides, and host cells comprising the polynucleotides and / or vectors. To express the desired polypeptide, a nucleotide sequence encoding anti-IL-11Rα, or a functional equivalent, may be inserted into a suitable expression vector, i.e., a vector that contains the necessary elements for the transcription and translation of the inserted coding sequence. Methods well known to those skilled in the art may be used to construct expression vectors containing a sequence encoding a polypeptide of interest and suitable transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Such techniques are described in Sambrook et al., Molecular Cloning, A Laboratory Manual (1989), and Ausubel et al., Current Protocols in Molecular Biology (1989).

[0177] A variety of expression vector / host systems are known and can be utilized to contain and express polynucleotide sequences, including, but not limited to, microorganisms, such as bacteria, transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors, yeast transformed with yeast expression vectors, insect cell systems infected with viral expression vectors (e.g., baculovirus), plant cell systems transformed with viral expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or with bacterial expression vectors (e.g., Ti or pBR322 plasmids), or animal cell systems, including mammalian cells, and more specifically, human cell systems.

[0178] "Control elements" or "regulatory sequences" present in an expression vector are those vector-enhancers, promoters, 5' and 3' untranslated regions that interact with host cell proteins to effect transcription and translation. Such elements may vary in their strength and specificity. Depending on the vector system and host utilized, any number of suitable transcription and translation elements may be used, including constitutive and inducible promoters. For example, when cloning in bacterial systems, inducible promoters may be used, such as the hybrid lacZ promoter of the PBLUESCRIPT phagemid (Stratagene, La Jolla, CA) or PSPORT1 plasmid (Gibco BRL, Gaithersburg, MD). In mammalian cell systems, promoters from mammalian genes or from mammalian viruses are generally preferred. When it is necessary to generate a cell line containing multiple copies of a polypeptide-encoding sequence, vectors based on SV40 or EBV may be advantageously used with appropriate selectable markers.

[0179] In bacterial systems, a number of expression vectors may be selected depending on the intended use for the expressed polypeptide. For example, where large quantities are required, vectors that direct high-level expression of fusion proteins that are easily purified may be used. Such vectors include, but are not limited to, multifunctional E. coli cloning and expression vectors, such as BLUESCRIPT (Stratagene), in which a sequence encoding a polypeptide of interest may be ligated into the vector in frame with a sequence for the amino-terminal Met and the following 7 residues of β-galactosidase, such that a hybrid protein is produced; pIN vectors (Van Heeke & Schuster, J. Biol. Chem. 264:5503 5509 (1989)); and the like. pGEX vectors (Promega, Madison, Wis.) may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). Generally, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Proteins made in such systems may be engineered to contain heparin, thrombin, or factor XA protease cleavage sites so that the cloned polypeptide of interest can, optionally, be released from the GST moiety.

[0180] Certain embodiments may use E. coli-based expression systems (see, e.g., Structural Genomics Consortium et al., Nature Methods. 5:135-146, 2008). These and related embodiments may rely partially or completely on ligation-independent cloning (LIC) to generate suitable expression vectors. In certain embodiments, protein expression may be controlled by T7 RNA polymerase (e.g., the pET vector series). These and related embodiments may utilize the expression host strain BL21(DE3), a lambda DE3 lysogen of BL21 that supports T7-mediated expression and is defective in lon and ompT proteases for improved target protein stability. Also included are expression host strains that carry plasmids encoding tRNAs that are rarely used in E. coli, such as ROSETTA™(DE3) and Rosetta 2(DE3) strains. Cell lysis and sample handling may also be improved using reagents sold under the trademarks BENZONASE® Nuclease and BUGBUSTER® Protein Extraction Reagent. For cell culture, autoinduction media can improve the efficiency of many expression systems, including high-throughput expression systems. This type of media (e.g., OVERNIGHT EXPRESS™ autoinduction system) gradually initiates protein expression through a metabolic shift without the addition of artificial inducers, such as IPTG. Certain embodiments use hexahistidine tags (such as those sold under the trademark HIS·TAG® fusions), followed by immobilized metal affinity chromatography (IMAC) purification, or related techniques. In certain aspects, however, clinical grade proteins can be isolated from E. coli inclusion bodies with or without the use of affinity tags (see, e.g., Shimp et al., Protein Expr Purif. 50:58-67, 2006). As a further example, certain embodiments may use a cold-shock inducible E. coli high-yield production system.This is because overexpression of proteins in E. coli at low temperatures improves their solubility and stability (see, e.g., Qing et al., Nature Biotechnology. 22:877-882, 2004).

[0181] Also included are high-density bacterial fermentation systems. For example, high cell density cultures of Ralstonia eutropha allow protein production at cell densities of over 150 g / L and expression of recombinant proteins at titers of over 10 g / L.

[0182] In the yeast Saccharomyces cerevisiae, numerous vectors containing constitutive or inducible promoters, such as alpha factor, alcohol oxidase, and PHH, can be used. For reviews, see Ausubel et al. (supra) and Grant et al., Methods Enzymol. 153:516-544 (1987). Also included is the Pichia pandoris expression system (see, e.g., Li et al., Nature Biotechnology. 24, 210-215, 2006; and Hamilton et al., Science, 301:1244, 2003). Certain embodiments include yeast systems engineered to selectively glycosylate proteins, including, inter alia, yeast with humanized N-glycosylation pathways (see, e.g., Hamilton et al., Science. 313:1441-1443, 2006; Wildt et al., Nature Reviews Microbiol. 3:119-28, 2005; and Gerngross et al., Nature-Biotechnology. 22:1409 -1414, 2004; U.S. Patent Nos. 7,629,163; 7,326,681; and 7,029,872). By way of example only, recombinant yeast cultures can be grown in Fernbach flasks or 15 L, 50 L, 100 L, and 200 L fermentors, among others.

[0183] In the case where a plant expression vector is used, expression of the polypeptide-encoding sequence can be driven by any of a number of promoters. For example, viral promoters such as the 35S and 19S promoters of CaMV can be used alone or in combination with the omega leader sequence from TMV (Takamatsu, EMBO J. 6:307-311 (1987)). Alternatively, plant promoters such as RUBISCO or the small subunit of the heat shock promoter can be used (Coruzzi et al., EMBO J. 3:1671-1680 (1984); Broglie et al., Science 224:838-843 (1984); and Winter et al., Results Probl. Cell Differ. 17:85-105 (1991)). These constructs can be introduced into plant cells by direct DNA transformation or pathogen-mediated transfection. Such techniques are described in many generally available reviews (see, e.g., Hobbs in McGraw Hill, Yearbook of Science and Technology, pp. 191-196 (1992)).

[0184] Insect systems can also be used to express a polypeptide of interest. For example, in one such system, Autographa californica nuclear polyhedrosis virus (AcNPV) is used as a vector to express foreign genes in Spodoptera frugiperda cells or in Trichoplusia cells. The polypeptide coding sequence may be cloned into a non-essential region of the virus, such as the polyhedrin gene, and placed under control of the polyhedrin promoter. Successful insertion of the polypeptide coding sequence renders the polyhedrin gene inactive and produces recombinant virus lacking coat protein. The recombinant virus may then be used to infect, for example, S. frugiperda or Trichoplusia cells in which the polypeptide of interest can be expressed (Engelhard et al., Proc. Natl. Acad. Sci. USA 91:3224-3227 (1994)). Also included are baculovirus expression systems, including those utilizing SF9, SF21, and T. ni cells (see, e.g., Murphy and Piwnica-Worms, Curr Protoc Protein Sci. Chapter 5:Unit 5.4, 2001). Insect systems can provide post-translational modifications similar to mammalian systems.

[0185] In mammalian host cells, many virus-based expression systems are commonly available. For example, when adenovirus is used as an expression vector, the sequence encoding the polypeptide of interest can be ligated into the adenovirus transcription / translation complex consisting of the late promoter and tripartite leader sequence. Insertion in the non-essential E1 or E3 region of the viral genome can be used to obtain a viable virus capable of expressing the polypeptide in infected host cells (Logan & Shenk, Proc. Natl. Acad. Sci. USA 81:3655-3659 (1984)). Transcription enhancers, such as the Rous sarcoma virus (RSV) enhancer, can also be used to increase expression in mammalian host cells.

[0186] Examples of useful mammalian host cell lines include the SV40 transformed monkey kidney CV1 line (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); mouse Sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); dog kidney cells (MDCK, ATCC CCL 34); buffalo rat hepatocytes (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TR1 cells (Mather et al., Annals NYAcad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; and human hepatoma lines (Hep G2). Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR-CHO cells (Urlaub et al., PNAS USA 77:4216 (1980)); and myeloma cell lines, such as NSO and Sp2 / 0. For a review of suitable specific mammalian host cell lines for antibody production, see, e.g., Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 255-268. Certain preferred mammalian cell expression systems include CHO and HEK293-cell based expression systems.Mammalian expression systems can utilize adherent cell lines, for example in T-flasks, roller bottles, or cell factories, or suspension cultures, for example, 1 L and 5 L spinners, 5 L, 14 L, 40 L, 100 L and 200 L stirred tank bioreactors, or 20 / 50 L and 100 / 200 L WAVE bioreactors, among others known in the art.

[0187] Also included is cell-free expression of proteins. These and related embodiments typically utilize purified RNA polymerase, ribosomes, tRNA, and ribonucleotides; these reagents may be produced by extraction from cells or from cell-based expression systems.

[0188] Specific initiation signals may also be used to achieve more efficient translation of sequences encoding a polypeptide of interest. Such signals include the ATG initiation codon and adjacent sequences. When a sequence encoding a polypeptide, its initiation codon, and upstream sequences are inserted into a suitable expression vector, additional transcriptional or translational control signals may not be required. However, when only a coding sequence, or a portion thereof, is inserted, exogenous translational control signals including the ATG initiation codon should be provided. Furthermore, the initiation codon should be in the correct reading frame to ensure translation of the entire insert. Exogenous translational elements and initiation codons can be of various origins, both natural and synthetic. The efficiency of expression can be enhanced by the inclusion of enhancers suitable for the particular cell system used, such as those described in the literature (Scharf.et al., Results Probl.Cell Differ.20:125-162 (1994)).

[0189] A host cell line may also be selected for its ability to regulate the expression of inserted sequences or process the expressed protein in a desired manner. Such modifications of polypeptides include, but are not limited to, post-translational modifications such as acetylation, carboxylation, glycosylation, phosphorylation, lipidation, and acylation. Post-translational processing that cleaves the "prepro" form of the protein may be used to facilitate correct insertion, folding, and / or function. Different host cells, such as yeast, CHO, HeLa, MDCK, HEK293, and W138, may be selected to ensure correct modification and processing of foreign proteins, in addition to bacterial cells that have or lack specific cellular and characteristic machinery for such post-translational activities.

[0190] For long-term, high-yield production of recombinant proteins, stable expression is generally preferred. For example, cell lines that stably express a polynucleotide of interest can be transformed using an expression vector that may contain a viral origin of replication and / or endogenous expression elements, as well as a selection marker gene on the same or a separate vector. After introduction of the vector, the cells may be grown in enriched medium for about 1-2 days before switching them to selective medium. The purpose of the selection marker is to confer resistance to selection, and its presence allows the growth and recovery of cells that successfully express the introduced sequence. Resistant clones of stably transformed cells may be propagated using tissue culture techniques appropriate for the cell type. Transient production, such as by transient transfection or infection, can also be used. Exemplary mammalian expression systems that are suitable for transient production include HEK293 and CHO-based systems.

[0191] Any number of selection systems may be used to recover transformed or transduced cell lines. These include, but are not limited to, the herpes simplex virus thymidine kinase (Wigler et al., Cell 11:223-232 (1977)) and adenine phosphoribosyltransferase (Lowy et al., Cell 22:817-823 (1990)) genes, which can be used in tk- or aprt- cells, respectively. Antimetabolite, antibiotic, or herbicide resistance can also be used as the basis for selection; for example, dhfr, which confers resistance to methotrexate (Wigler et al., PNAS USA. 77:3567-70 (1980)); npt, which confers resistance to aminoglycosides, neomycin, and G-418 (Colbere-Garapin et al., J. Mol. Biol. 150:1-14 (1981)); and als or pat, which confer resistance to chlorsulfuron and phosphinothricin acetyltransferase, respectively (Murry, supra). Additional selectable genes have been described, such as trpB, which allows cells to utilize indole instead of tryptophan, or hisD, which allows cells to utilize histinol instead of histidine (Hartman & Mulligan, Proc. Natl. Acad. Sci. USA 85:8047-51 (1988)). The use of visible markers has gained popularity with markers such as green fluorescent protein (GFP) and other fluorescent proteins (e.g., RFP, YFP), anthocyanins, β-glucuronidase and its substrate GAS, and luciferase and its substrate luciferin being widely used not only to identify transformants, but also to quantify the amount of transient or stable protein expression resulting from a particular vector system (see, e.g., Rhodes et al., Methods Mol. Biol. 55:121-131 (1995)).

[0192] Also included are high throughput protein production systems, or microproduction systems. Certain embodiments may utilize hexahistidine fusion tags for protein expression and purification, for example, on metal chelate modified slide surfaces or MagneHis Ni-particles (see, e.g., Kwon et al., BMC Biotechnol. 9:72, 2009; and Lin et al., Methods Mol Biol. 498:129-41, 2009). Also included are high throughput cell-free protein expression systems (see, e.g., S Sitaraman et al., Methods Mol Biol. 498:229-44, 2009). These and related embodiments can be used, for example, to generate microarrays of antibodies, which can then be used to screen libraries to identify antibodies and antigen binding domains that interact with the IL-11Rα polypeptide of interest.

[0193] A variety of protocols for detecting and measuring expression of polynucleotide-encoded products using binding agents or antibodies, such as polyclonal or monoclonal antibodies specific for the product, are known in the art. Examples include enzyme-linked immunosorbent assay (ELISA), Western immunoblot, radioimmunoassay (RIA), and fluorescence-activated cell sorting (FACS). These and other assays are described, inter alia, in Hampton et al., Serological Methods, a Laboratory Manual (1990) and Maddox et al., J. Exp. Med. 158:1211-1216 (1983).

[0194] A wide variety of labeling and conjugation techniques are known to those skilled in the art and can be used in various nucleic acid and amino acid assays. Means for producing labeled hybridization or PCR probes for detecting sequences related to polynucleotides include oligo-labeling, nick translation, end-labeling, or PCR amplification using labeled nucleotides. Alternatively, the sequence, or any portion thereof, may be cloned into a vector for the production of an mRNA probe. Such vectors are known in the art and commercially available and can be used to synthesize RNA probes in vitro by adding a suitable RNA polymerase, such as TT7, T3, or SP6, and labeled nucleotides. These procedures may be performed using a variety of commercially available kits. Suitable reporter molecules or labels that can be used include radionuclides, enzymes, fluorescent, chemiluminescent, or chromogenic agents as well as substrates, cofactors, inhibitors, magnetic particles, and the like.

[0195] Host cells transformed with a polynucleotide sequence of interest may be cultured under conditions appropriate for the expression and recovery of the protein from cell culture. Certain embodiments utilize serum-free cell expression systems. Examples include HEK293 cells and CHO cells, which can be grown on serum-free media (see, e.g., Rosser et al., Protein Expr. Purif. 40:237-43, 2005; and U.S. Patent No. 6,210,922).

[0196] Antibodies, or antigen-binding fragments thereof, produced by recombinant cells may be secreted or contained intracellularly depending on the sequence and / or vector used. As will be appreciated by those skilled in the art, expression vectors containing polynucleotides may be designed to contain signal sequences that direct secretion of the encoded polypeptide through a prokaryotic or eukaryotic cell membrane. Other recombinant constructs may be used to link sequences encoding a polypeptide of interest to nucleotide sequences encoding polypeptide domains that facilitate purification and / or detection of soluble proteins. Examples of such domains include cleavable and non-cleavable affinity purification tags and epitope tags, such as avidin, FLAG tags, polyhistidine tags (e.g., 6xHis), cMyc tags, V5 tags, glutathione S-transferase (GST) tags, and others.

[0197] Proteins produced by recombinant cells can be purified and characterized according to a variety of techniques known in the art. Exemplary systems for performing protein purification and analyzing protein purity include fast protein liquid chromatography (FPLC) (e.g., AKTA and Bio-Rad FPLC systems), high pressure liquid chromatography (HPLC) (e.g., Beckman and Waters HPLC). Exemplary chemistries for purification include ion exchange chromatography (e.g., Q, S), size exclusion chromatography, salt gradients, affinity purification (e.g., Ni, Co, FLAG, maltose, glutathione, protein A / G), gel filtration, reverse phase, ceramic HYPERD® ion exchange chromatography, and hydrophobic interaction columns (HIC), among others, as known in the art. Also included are analytical methods, such as SDS-PAGE (e.g., Coomassie, silver stain), immunoblot, Bradford, and ELISA, which may typically be utilized during any step of the production or purification process to measure the purity of a protein composition.

[0198] Also included are methods for concentrating anti-IL-11Rα antibodies and antigen-binding fragments thereof, and compositions comprising the concentrated soluble protein. In certain embodiments, concentrated solutions of anti-IL-11Rα antibodies comprise a protein concentration of about 5 mg / mL; or about 8 mg / mL; or about 10 mg / mL; or about 15 mg / mL; or about 20 mg / mL or more.

[0199] In some aspects, the compositions are substantially monodisperse, e.g., where the anti-IL-11Rα antibodies are present predominantly (i.e., at least about 90% or more) in one apparent molecular weight form, as assessed, e.g., by size exclusion chromatography, dynamic light scattering, and / or analytical ultracentrifugation.

[0200] In some aspects, the compositions have at least about 90%, or in some aspects at least about 95% purity, or in some embodiments at least about 98% purity (protein basis). Purity can be determined via any routine analytical method known in the art.

[0201] In some embodiments, the composition has a high molecular weight aggregate content of less than about 10%, less than about 5%, less than about 3%, or less than about 1%. High molecular weight aggregate content can be determined by a variety of analytical techniques, including, for example, size exclusion chromatography, dynamic light scattering, and / or analytical ultracentrifugation.

[0202] Examples of concentration approaches contemplated herein include lyophilization, which is typically used when the solution contains few soluble components other than the protein of interest. Lyophilization is often performed after an HPLC run and can remove most or all volatile components from the mixture. Also included are ultrafiltration techniques, which typically use one or more selectively permeable membranes to concentrate protein solutions. The membrane allows water and small molecules to pass through and retain the protein; the solution can be forced against the membrane by mechanical pumps, gas pressure, or centrifugation, among other techniques.

[0203] In certain embodiments, the anti-IL-11Rα antibodies, reagents, or related agents have a purity of at least about 90% as measured according to routine techniques in the art. In certain embodiments, the anti-IL-11Rα compositions have a purity of at least about 95%. In certain embodiments, such as therapeutic or pharmaceutical compositions, the anti-IL-11Rα antibody compositions have a purity of at least about 97% or 98% or 99%. In some embodiments, such as when used as a reference or research reagent, the anti-IL-11Rα antibodies may be less pure and have a purity of at least about 50%, 60%, 70%, or 80%. Purity can be measured overall or in relation to selected components, such as other proteins, e.g., purity on a protein basis.

[0204] The purified antibodies can also be characterized according to their biological characteristics. Binding affinity and binding kinetics can be measured according to various techniques known in the art, such as Biacore® and related techniques utilizing surface plasmon resonance (SPR), an optical phenomenon that allows detection of unlabeled interactors in real time. SPR-based biosensors can be used in determining activity concentration, screening, and characterization, both in terms of affinity and kinetics. The presence or level of one or more reference or non-reference biological activities can be measured according to cell-based assays, including assays utilizing cellular binding partners of selected anti-IL-11Rα antibodies functionally coupled to a readout or indicator, such as a fluorescent or luminescent indicator of biological activity, as described herein.

[0205] In certain embodiments, the composition, as described above, is substantially endotoxin-free, for example, about or at least about 95% endotoxin-free, about or at least about 99% endotoxin-free, or about or at least about 99.99% endotoxin-free. The presence of endotoxin can be detected according to routine techniques in the art, as described herein. In certain embodiments, the composition is made from eukaryotic cells, such as mammalian or human cells, in a substantially serum-free medium. In certain embodiments, the composition, as described herein, has an endotoxin content of less than about 10 EU / mg antibody, or less than about 5 EU / mg antibody, or less than about 3 EU / mg antibody, or less than about 1 EU / mg antibody.

[0206] In certain embodiments, the composition comprises less than about 10% w / w high molecular weight aggregates, or less than about 5% w / w high molecular weight aggregates, or less than about 2% w / w high molecular weight aggregates, or less than about 1% w / w high molecular weight aggregates.

[0207] Also included are protein-based analytical assays and methods that can be used to assess, for example, protein purity, size, solubility, and degree of aggregation, among other characteristics. Protein purity can be assessed in a number of ways. For example, purity can be assessed based on primary structure, higher order structure, size, charge, hydrophobicity, and glycosylation. Examples of methods for assessing primary structure include N- and C-terminal sequencing and peptide mapping (see, e.g., Allen et al., Biologicals. 24:255-275, 1996). Exemplary methods for assessing conformation include circular dichroism (see, e.g., Kelly et al., Biochim Biophys Acta. 1751:119-139, 2005), fluorescence spectroscopy (see, e.g., Meagher et al., J. Biol. Chem. 273:23283-89, 1998), FT-IR, amide hydrogen-deuterium exchange kinetics, differential scanning calorimetry, NMR spectroscopy, and immunoreactivity with conformation-sensitive antibodies. Conformation can also be assessed as a function of various parameters, such as pH, temperature, or added salt. Exemplary methods for assessing protein characteristics, such as size, include analytical ultracentrifugation and size-exclusion HPLC (SEC-HPLC), and exemplary methods for measuring charge include ion-exchange chromatography and isoelectric focusing. Hydrophobicity can be assessed, for example, by reversed-phase HPLC and hydrophobic interaction chromatography HPLC. Glycosylation can affect pharmacokinetics (eg, clearance), conformation or stability, receptor binding, and protein function and can be assessed, for example, by mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy.

[0208] As noted above, certain embodiments include the use of SEC-HPLC to evaluate protein characteristics, such as purity, size (e.g., size homogeneity), or degree of aggregation, among other uses, and / or to purify proteins. SEC also includes gel filtration chromatography (GFC) and gel permeation chromatography (GPC) and refers to a chromatographic method in which molecules in a solution are separated in a porous material based on their size, or more specifically, their hydrodynamic volume, diffusion coefficient, and / or surface properties. This process is generally used to separate biological molecules and to determine the molecular weight and molecular weight distribution of polymers. Typically, a biological or protein sample (such as a protein extract produced according to the protein expression methods provided herein and known in the art) is loaded into a selected size exclusion column with a defined stationary phase (porous material), preferably a phase that does not interact with the proteins in the sample. In certain aspects, the stationary phase is composed of inert particles packed in a dense three-dimensional matrix within a glass or steel column. The mobile phase can be pure water, an aqueous buffer, an organic solvent, or a mixture thereof. Stationary phase particles typically have small pores and / or channels that allow only molecules below a certain size to enter. Larger particles are therefore excluded from these pores and channels, and their limited interaction with the stationary phase leads them to elute as a "totally excluded" peak at the beginning of the experiment. Smaller molecules can fit into the pores and are removed from the flowing mobile phase, and the time they spend immobilized in the stationary phase pores depends, in part, on how far into the pores they penetrate. Their removal from the mobile phase flow causes them to elute for a longer time from the column, resulting in a separation between particles based on differences in their size. A given size exclusion column has a range of molecular weights that can be separated. Overall, molecules larger than the upper limit are not captured by the stationary phase, molecules smaller than the lower limit enter the solid phase completely and elute as a single band, and molecules within the range elute at different rates defined by their properties, such as hydrodynamic volume.For examples of these methods in practice with pharmaceutical proteins, see Bruner et al., Journal of Pharmaceutical and Biomedical Analysis. 15: 1929-1935, 1997.

[0209] Protein purity for clinical applications is discussed, for example, by Anicetti et al. (Trends in Biotechnology. 7:342-349, 1989). More recent techniques for analyzing protein purity include, but are not limited to, the LabChip GXII, an automated platform for rapid analysis of proteins and nucleic acids, which provides high-throughput analysis of protein titer, size, and purity analysis. In certain non-limiting embodiments, clinical grade proteins, such as protein fragments and antibodies, can be obtained by utilizing a combination of chromatographic materials in at least two orthogonal steps, among other methods (see, e.g., Therapeutic Proteins: Methods and Protocols. Vol. 308, Eds., Smales and James, Humana Press Inc., 2005). Typically, protein drugs (e.g., antibodies and antigen-binding fragments) are substantially free of endotoxins, as measured according to techniques known in the art and described herein.

[0210] Protein solubility assays are also included. Such assays can be utilized, for example, to determine optimal growth and purification conditions for recombinant production, to optimize buffer selection, and to optimize antibody or antigen-binding fragment selection. Solubility or aggregation can be assessed according to a variety of parameters, including temperature, pH, salt, and the presence or absence of other additives. Examples of solubility screening assays include, without limitation, microplate-based methods that measure protein solubility using turbidity or other measures as an endpoint, high-throughput assays for analysis of purified recombinant protein solubility (see, e.g., Stenvall et al., Biochim Biophys Acta. 1752:6-10, 2005), assays that use structural complementation of genetic marker proteins to monitor and measure protein folding and solubility in vivo (see, e.g., Wigley et al., Nature Biotechnology. 19:131-136, 2001), and electrochemical screening of recombinant protein solubility in E. coli using scanning electrochemical microscopy (SECM) (see, e.g., Nagamine et al., Biotechnology and Bioengineering. 96:1008-1013, 2006), among others. Antibodies with increased solubility (or reduced aggregation) can be identified or selected according to routine techniques in the art, including simple in vivo assays for protein solubility (see, e.g., Maxwell et al., Protein Sci. 8:1908-11, 1999).

[0211] Protein solubility and aggregation can also be measured by dynamic light scattering techniques. Aggregation is a general term that encompasses several types of interactions or characteristics, including soluble / insoluble, covalent / non-covalent, reversible / irreversible, and native / denatured interactions and characteristics. For protein therapeutics, the presence of aggregates is typically considered undesirable due to concerns that aggregates may cause immunogenic reactions (e.g., small aggregates) or adverse events upon administration (e.g., particulates). Dynamic light scattering refers to a technique that can be used to determine the size distribution profile of small particles in suspension or polymers in solution, such as proteins. This technique, also referred to as photon correlation spectroscopy (PCS) or quasi-elastic light scattering (QELS), uses scattered light to measure the rate of diffusion of protein particles. Variations in scattering intensity can be observed due to Brownian motion of molecules and particles in solution. This motion data can be conventionally processed to derive a size distribution for the sample, in which the size is given by the Stokes radius or hydrodynamic radius of the protein particles. Hydrodynamic size depends on both mass and shape (composition). Dynamic scattering can detect the presence of very small amounts of aggregated protein (<0.01% by weight) even in samples containing a wide range of masses. It can also be used to compare the stability of different formulations, including applications that rely on real-time monitoring of changes at elevated temperatures. Thus, certain embodiments include the use of dynamic light scattering to analyze the solubility and / or presence of aggregates in samples containing the antibodies of the present disclosure.

[0212] Although the foregoing embodiments have been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to one of ordinary skill in the art in view of the teachings of the present disclosure that certain changes and modifications may be made without departing from the spirit or scope of the appended claims. The following examples are provided by way of illustration only, and not by way of limitation. Those of ordinary skill in the art will readily recognize a variety of non-critical parameters that can be changed or modified to yield essentially similar results. EXAMPLES

[0213] Example 1 Anti-IL-11Rα antibody Tests were conducted to generate improved anti-IL-11Rα antibodies compared to the antibodies described in U.S. Patent Nos. 9,796,782 and 9,340,618, including, for example, the antibodies designated 8E2 and TS7. For example, the V of the TS7 antibody L Potential N-linked glycosylation in CDR3 was removed to reduce heterogeneity and improve developability, and other sequence changes were made to improve binding affinity and potency. The CDR sequences of the antibodies are provided in Table A1 (above). Antibodies were expressed in HEK293 cells and purified using Protein A-based chromatography using standard methodologies. All antibodies were expressed in human IgG4 (S228P according to EU numbering; S241P according to Kabat numbering) format with kappa light chains.

[0214] The relative binding activity of the antibodies was tested by competitive ELISA using biotinylated mAb 8E2 or TS7. Antibodies 8E2 and TS7 were biotinylated using sulfo-NHS biotin (Thermo Fisher) according to the manufacturer's instructions. IL-11R was coated onto an ELISA plate (Nunc, MaxiSorp) at 2 μg / mL in PBS, and the plate was blocked with 2% BSA in PBS. After washing the plate with water, the antibody to be tested was titrated across the plate in a final volume of 90 μL, followed by the addition of 10 μL of biotinylated 8E2 or biotinylated TS7 to each well (final concentration of 0.1 μg / mL) as shown in Table E1. The plate was incubated for 2 hours on a plate shaker, followed by washing and the addition of 100 μL / well of streptavidin-HRP (Jackson ImmunoResearch, 1 / 1000 dilution). After a further 1 hour incubation, plates were washed and developed with Ultra-TMB HRP substrate (Thermo Fisher) and stopped by the addition of 50 μL / well of 2 M sulfuric acid. Plates were read at 450 nm on an iD5 plate reader (Molecular Devices).

[0215] As shown in Table E1 below, many of the antibodies tested were shown to have increased binding to human IL-11Rα compared to the binding of the 8E2 and TS7 antibodies. [Table 5]

[0216] Kinetic binding analysis of mAbs was tested to determine antigen binding affinity using biolayer interferometry on an Octet RED96e instrument. mAbs were loaded onto a protein G biosensor (ForteBio) in 10x kinetic buffer consisting of PBS, 0.1% BSA, 0.02% Tween 20 for 120 seconds, yielding spectral shift values ​​of 0.8-1.2 nm. Association was performed in the presence of a two-fold dilution series of hIL-11Rα and allowed to proceed for 120 seconds; dissociation was measured over 300-1200 seconds. The dilution series started at 100 nM for the weaker variants or 10 nM for the most potent mAb.

[0217] As shown in Table E2 below, mAb5, mAb6, mAb7, mAb8, mAb9, mAb12, mAb13, mAb14, and mAb15 had extremely high binding affinities that exceeded the ability of the instrument to determine accurate off-rates (kd). [Table 6]

[0218] Further testing of the lead mAbs was performed using a cell-based reporter assay. IL-11 has been reported to signal through the STAT3 and ERK pathways. Therefore, a STAT3 reporter cell line in which firefly luciferase gene expression is driven by a STAT3 response element located upstream of a minimal TATA promoter was selected to analyze mAb functional activity. In these cells, IL-11 can activate endogenous STAT3, which can bind to the endogenous STAT3 response element and induce transcription of the luciferase reporter gene, resulting in easily detectable luciferase expression. Antibodies against IL-11R that block IL-11 signaling should inhibit IL-11-driven luciferase expression in these cells.

[0219] STAT3 reporter (luciferase)-HEK293 cell line (BPS Bioscience, Cat. No. 79800-P) was grown, passaged, and assayed according to the manufacturer's protocol. Cells were plated at 25,000 cells per well in 96-well microtiter dishes and incubated at 37 °C for 24 h at 5% CO. 2 Plates were incubated for 30-45 minutes at 37°C in humidified air. Anti-IL-11R mAb was added as a 3-fold dilution series in duplicate columns or rows and the plates were returned to the incubator for an additional hour at 37°C, after which 40ng / mL IL-11 was added to each well to initiate the signaling cascade and luciferase production. Plates were incubated for 18-24 hours at 37°C and luciferase was detected using the One-Step Luciferase Assay System (BPS Bioscience, Cat. No. 60690-1) according to the manufacturer's instructions.

[0220] As shown in Table E3, TS7 is significantly more potent than 8E2, and mAb5, mAb9, and mAb13 are more potent at inhibiting IL-11 signaling than either TS7 or 8E2. Consistent with the ELISA data, mAb7, mAb8, mAb10, and mAb11 were less potent than TS7, and mAb12, mAb14, and mAb15 were comparable in potency to TS7. Results for mAb6 were variable in this assay, but the ELISA data and Octet data above suggest that mAb6 is also a potent mAb. [Table 7]

[0221] A further series of antibodies was designed in which the residue at Kabat position 31 of the heavy chain was varied (mAb16-32). In this case, an alternative biolayer interferometry assay was used to first rank the antibodies relative to mAb5, the closest in sequence to mAb16-32. MAbs were loaded onto an anti-human constant domain (AHC) biosensor (ForteBio) in 10x kinetic buffer consisting of PBS, 0.1% BSA, 0.02% Tween 20 for 120 seconds, achieving spectral shift values ​​of 0.8-1.2 nm. Association was performed in the presence of a two-fold dilution series of hIL-11R and allowed to proceed for 120 seconds. The dilution series started at 10 nM. Dissociation in 10x kinetic buffer for 1200 seconds. Binding kinetic measurements were taken on an Octet RED96e instrument.

[0222] As shown in Table E4, initial qualitative assessment identified additional antibodies with high potency similar to or better than mAb5. More detailed kinetic measurements were obtained for mAb21, mAb23, mAb29, and mAb31 and compared to mAb5, thereby confirming further improved potency for these mAbs. Further testing of these mAbs by ELISA confirmed the improved potency of selected mAbs. [Table 8]

[0223] All publications, patent applications, and issued patents cited in this specification are herein incorporated by reference to the same extent as if each individual publication, patent application, or issued patent was specifically and individually indicated to be incorporated by reference.

[0224] Example 2. Activity of anti-IL-11Rα antibodies in a fibrotic human precision-cut lung slice model A study was conducted to evaluate the effect of the mAb5 antibody on biomarkers reflecting ECM remodeling in a fibrotic human precision-cut lung slice (PCLS) model. The anti-fibrotic effect was examined at three different concentrations in human ex vivo lung tissue slices prepared from patients with pulmonary fibrosis.

[0225] Materials and Methods Tissue slices were prepared from the subpleural and central regions of fibrotic lungs from two human end-stage donors with pulmonary fibrosis (IPF and secondary fibrosis) (see, e.g., Hess et al., Toxicol In Vitro. 32:347-61, 2016). For both donors, lung tissue samples from two different regions (subcutaneous and central) were used to generate tissue cores used for PCLS. Adjacent cores were used to achieve sequential PCLS for analysis of one treatment compared to medium-treated PCLS from the same core. This allows comparison of densely packed fibrotic areas with fibrotic foci to less affected areas. Clinical / demographic information for each patient is provided to ensure fibrotic status.

[0226] The experimental treatments included are listed in Table E5. Each treatment was tested in three replicates for each lung region and for each donor. [Table 9]

[0227] Preparation and incubation of precision-cut lung slices. Lung lobes were cannulated and filled with warm 2% low melting agarose / medium solution at 37°C. The filled lobes were cooled on ice to allow the agarose to polymerize. Tissue cores with a diameter of 8 mm were prepared and cut into slices approximately 300 μm thick using a microtome (Krumdieck tissue slicer, Alabama Research and Development, Mumford, AL, USA, or Vibratome OTS-5000, Science Services GmbH, Munich) in Earle's Balanced Salts Solution (EBSS). Tissue slices were incubated in Dulbecco's Modified Eagle's Medium / Nutrient Mixture F-12 Ham (DMEM) without phenol red, with L-glutamine and 15 mM HEPES. Tissue slices were transferred into Petri dishes and washed with DMEM for 2 h. The medium was changed four times every 30 min to remove cell debris.

[0228] Two PCLS per well were left in 500 μL of DMEM in a 24-well plate overnight for a final washing step. The next day, the PCLS were cultured under normal cell culture conditions (37° C., 5% CO 2 ) for 48 hours in 250 μl of DMEM / F-12 medium with experimental treatments or controls (see Table E5). Medium contained 100 units / mL penicillin and 100 μg / mL streptomycin but was not supplemented with fetal bovine serum.

[0229] After 48 h of incubation, the supernatants were collected, proteinase inhibitor cocktail (0.2%) was added, and samples were frozen to −80° C. In addition, two PCLS from each well were transferred to 2 mL safelock tubes, immediately flash frozen in liquid nitrogen, and stored at −80° C. Samples from the three replicates for each condition were not pooled.

[0230] Tissue viability and responsiveness to immune stimulants were confirmed in quality control samples in which PCLS were incubated with the mitogen lipopolysaccharide (LPS) at 100 ng / mL. After 48 h of incubation, assessment of viability by lactate dehydrogenase (LDH) activity and pro-inflammatory responses by IL-1β were performed.

[0231] Tissue viability Lung tissue viability was evaluated to ensure sufficient quality of PCLS. For quality control samples, LDH activity was determined with a commercial enzyme assay in the supernatant after incubation. An increase in the amount of dead or membrane-damaged cells in PCLS leads to an increase in LDH enzyme activity in the culture supernatant. Triton X-100 (1% in PBS)-treated PCLS was examined as a reference control. 50 μL of sample was incubated with 50 μL of LDH reagent for 20 min at room temperature. Absorbance was measured at 490 nm and 630 nm as reference wavelengths using a microplate reader. All tests were performed as a single measurement per well. Due to the dense tissue material of the fibrous PCLS, the absorbance value measured for the Triton X-100 control was the highest. Therefore, a 1:10 dilution of these controls was tested in parallel to achieve a suitable value within the detection spectrum for the calculation of the relative LDH release of the samples.

[0232] Tissue responsiveness The general responsiveness of lung tissue to treatment was evaluated to ensure optimal condition of PCLS. For quality control samples, PCLS were lysed with Triton X-100 (1% in PBS) for 1 hour, after which endogenous IL-1β levels were assessed using an ELISA from R&D according to the manufacturer's instructions. Photometric measurements were performed at 450 nm and a reference wavelength of 570 nm using a microplate reader. IL-1β concentrations were normalized to the total protein concentration of PCLS determined by BCA assay.

[0233] Tissue Appearance The macroscopic appearance of the tissues used to generate PCLS as well as the location of the cores were photographically documented. Tissue cores adjacent to those used for PCLS preparation were fixed in 10% formalin and subsequently embedded in paraffin. Paraffin-embedded tissue sections were further processed into slices for staining with hematoxylin and eosin (H&E) to assess fibrotic remodeling.

[0234] Extracellular Matrix Remodeling Assessment Biomarkers reflecting ECM remodeling were assessed in the supernatants by Nordic Bioscience using the following competitive ELISA: PRO-C6 (incorporating the C-terminus of collagen VI C5 domain released as collagen VI into the matrix; measured on a mesoscale platform [hsPRO-C6]): reflects collagen VI formation. Collagen VI is a beaded filament collagen found primarily at the interface between the interstitial matrix and the basement membrane.

[0235] Statistics Biomarker data are presented as scatter plots with indication of median and interquartile range (IQR). Data are presented as raw biomarker values ​​for each region and donor. Pooled data are presented as percentages of "no treatment" controls, calculated for individual regions and donors. Statistical significance was assessed on pooled data by Kruskal-Wallis using Dunn's multiple comparison test compared to "no treatment" controls. Adjusted p-values ​​are indicated as *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0236] result Fibrotic Conditions Human PCLS were generated from lung tissue from two donors with pulmonary fibrosis. Pathology reports provided after completion of the study noted that the fibrotic tissue displayed a distinct UIP pattern, compatible with IPF (see Table E6). [Table 10]

[0237] Tissue Appearance PCLS were prepared from different regions of the fibrotic lung. Figures 1A-1B show images of each donor's lung and the area used, where the drawn circle marks the area of ​​tissue used to generate PCLS. Adjacent tissue cores from the same lung areas used for PCLS were then prepared for H&E staining to assess fibrotic remodeling of the tissue (see Figures 2A-2B). Donor 2 appeared to have more extensive tissue changes than Donor 1, which is in accordance with the statement in the pathology report that Donor 2 tissue is compatible with IPF in a much more advanced stage.

[0238] Tissue viability Tissue viability of quality control samples (LPS) was measured by LDH release after 48 hours of incubation. LDH release <20% of the Triton X lysis control was observed for all samples, indicating that tissue was viable over the 48 hour experimental period (see Figures 3A-3B).

[0239] Tissue responsiveness Tissue responsiveness of quality control samples (LPS) was analyzed by IL-1β expression. Analysis showed that human fibrotic lung tissue from both donors responded to the pro-inflammatory stimulus LPS, identified by upregulated IL-1β expression after 48 hours (see Figures 4A-4B).

[0240] Extracellular matrix remodeling Collagen VI formation was assessed by PRO-C6. Media background levels of PRO-C6 were low and at the lower limit of detection for the assay. Donor 1 showed lower levels of PRO-C6 compared to donor 2, with levels much closer to the moderate background.

[0241] As shown in Figure 5, pooled PRO-C6 data for all donors and regions showed that the median levels of PRO-C6 for nintedanib and all concentrations of mAb5 (anti-IL11R) were decreased compared to the untreated (medium) control. mAb5 reduced PRO-C6 in a dose-dependent manner. The isotype control showed no change in median PRO-C6 levels compared to the untreated (medium) control, and even tended to increase slightly.

[0242] Based on the effect of nintedanib, lung slices were responsive to antifibrotic treatment because ECM remodeling could be reduced. Overall, a trend towards an effect on ECM remodeling was observed with the mAb5 anti-IL-11Rα antibody. Of particular interest was the dose-related reduction in collagen VI formation (PRO-C6) by the mAb5 antibody. In some cases, the mAb5 antibody showed a better effect on ECM remodeling than the approved treatment nintedanib, strengthening the conclusion that this antibody has an antifibrotic effect.

Claims

1. 1. An isolated antibody, or antigen-binding fragment thereof, that binds to interleukin-11 receptor subunit alpha (IL-11Rα), wherein said at least one antibody or antigen-binding fragment thereof is: Complementarity-determining region V Ｈ CDR1, V Ｈ CDR2, and V Ｈ The heavy chain variable region (V) containing the CDR3 sequence Ｈ ) and Complementarity-determining region V Ｌ CDR1, V Ｌ CDR2, and V Ｌ The light chain variable region (V) containing the CDR3 sequence Ｌ ), a) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 1 to 3, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 4-6, respectively; b) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 7 to 9, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 10-12, respectively; c) Said V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 13 to 15, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 16-18, respectively; d) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 19 to 21, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 22-24, respectively; e) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 25-27, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 28-30, respectively; f) Said V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 31 to 33, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 34-36, respectively; g) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 37-39, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 40-42, respectively; h) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 43-45, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 46-48, respectively; i) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 49 to 51, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 52-54, respectively; j) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 55-57, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 58-60, respectively; k) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 61-63, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 64-66, respectively; l) Said V Ｈ CDR1, V Ｈ CDR2, and V Ｈ the CDR3 sequences comprise SEQ ID NOs: 67-69, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 70-72, respectively; m) Said V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 73-75, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 76-78, respectively; n) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 79-81, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 82-84, respectively; o) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 85-87, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 88-90, respectively; p) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 91-93, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 94-96, respectively; q) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 97-99, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 100-102, respectively; r) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 103-105, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 106-108, respectively; s) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ the CDR3 sequences comprise SEQ ID NOs: 109-111, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 112-114, respectively; t) Said V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 115-117, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 118-120, respectively; u) Said V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 121-123, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 124-126, respectively; v) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ the CDR3 sequences comprise SEQ ID NOs: 127-129, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 130-132, respectively; w) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ the CDR3 sequences comprise SEQ ID NOs: 133-135, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 136-138, respectively; x) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 139-141, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 142-144, respectively; y) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 145-147, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 148-150, respectively; z) Said V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 151-153, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 154-156, respectively; aa) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 157-159, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 160-162, respectively; bb) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ the CDR3 sequences comprise SEQ ID NOs: 163-165, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 166-168, respectively; cc) Said V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 169-171, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 172-174, respectively; dd) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ the CDR3 sequences comprise SEQ ID NOs: 175-177, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ CDR3 sequences comprise SEQ ID NOs: 178-180, respectively; ee) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ CDR3 sequences comprise SEQ ID NOs: 181-183, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ the CDR3 sequences comprise SEQ ID NOs: 184-186, respectively; or ff) the V Ｈ CDR1, V Ｈ CDR2, and V Ｈ the CDR3 sequences comprise SEQ ID NOs: 187-189, respectively, and Ｌ CDR1, V Ｌ CDR2, and V Ｌ An isolated antibody, or antigen-binding fragment thereof, wherein the CDR3 sequences comprise SEQ ID NOs: 190-192, respectively.

2. 2. The isolated antibody, or antigen-binding fragment thereof, of claim 1, a) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 193, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 194; b) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 195, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 196; c) Said V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 197, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 198; d) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 199, and said V Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 200; e) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 201, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 202; f) Said V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 203, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 204; g) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 205, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 206; h) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 207, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 208; i) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 209, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 210; j) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 211, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 212; k) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 213, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 214; l) Said V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 215, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 216; m) Said V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 217, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 218; n) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 219, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 220; o) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 221, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 222; p) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 223, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 224; q) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 225, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 226; r) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 227, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 228; s) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 229, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 230; t) Said V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 231, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 232; u) Said V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 233, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 234; v) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 235, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 236; w) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 237, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 238; x) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 239, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 240; y) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 241, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 242; z) Said V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 243, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 244; aa) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 245, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 246; bb) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 247, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 248; cc) Said V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 249, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 250; dd) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 251, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 252; ee) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 253, and Ｌ comprises a sequence at least 80% identical to SEQ ID NO: 254; or ff) the V Ｈ comprises a sequence at least 80% identical to SEQ ID NO: 255, and Ｌ An isolated antibody, or antigen-binding fragment thereof, comprising a sequence at least 80% identical to SEQ ID NO:

256.

3. 2. The isolated antibody of claim 1, or an antigen-binding fragment thereof, which binds to fibronectin domain III of human IL-11Rα, or to about residues 112-219 of SEQ ID NO:

257.

4. 10. The isolated antibody, or antigen-binding fragment thereof, of claim 1, having one or more of the following characteristics: have a binding affinity for human IL-11Rα of less than about 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 40, or 30 pM, and optionally have an increased binding affinity for human IL-11Rα compared to that of the TS7 and 8E2 antibodies; antagonizing the binding and / or signaling activity between IL-11Rα and IL-11, optionally with increased potency as an IL-11 signaling antagonist compared to that of the TS7 and 8E2 antibodies; optionally reducing IL-11Rα / gp130 dimerization or complex formation in a cell-based assay; and / or In some cases, V compared to TS7 and 8E2 antibodies Ｌ An isolated antibody, or antigen-binding fragment thereof, having reduced N-linked glycosylation in CDR3.

5. 2. The isolated antibody, or antigen-binding fragment thereof, of claim 1, comprising an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or a hybrid, variant, or modified Fc domain thereof.

6. 2. The isolated antibody, or antigen-binding fragment thereof, of claim 1, comprising a modified IgG Fc domain with enhanced effector function in humans compared to a wild-type or native Fc region, optionally wherein the modified IgG Fc domain is an IgG1 or IgG3 Fc domain.

7. 2. The isolated antibody, or antigen-binding fragment thereof, of claim 1, comprising an IgG Fc domain with reduced effector function in humans compared to a wild-type or native Fc region, optionally wherein the modified IgG Fc domain is an IgG2 or IgG4 Fc domain.

8. 2. The isolated antibody, or antigen-binding fragment thereof, of claim 1, comprising a human IgG1 or IgG4 Fc domain, or a hybrid, variant, or modified Fc domain thereof, optionally wherein the human IgG4 Fc domain comprises SEQ ID NO: 258 or 259.

9. 2. The isolated antibody or antigen-binding fragment thereof of claim 1, The antibody comprises an altered Fc region that exhibits any one or more of the following: Increased half-life, decreased complement fixation or activation, decreased Clq binding affinity, decreased CDC-related activity, decreased ADCC-related activity, and decreased ADCP-related activity compared to the corresponding wild-type Fc sequence.

10. 2. The isolated antibody, or antigen-binding fragment thereof, of claim 1, which is a monoclonal antibody.

11. is a humanized antibody, 2. The isolated antibody, or antigen-binding fragment thereof, of claim 1, optionally wherein the antibody, or antigen-binding fragment thereof, is a humanized monoclonal antibody comprising a human IgG4 Fc domain with a S228P mutation (EU numbering).

12. 2. The isolated antibody, or antigen-binding fragment thereof, of claim 1, selected from an Fv fragment, a single-chain Fv (scFv) polypeptide, an adnectin, an anticalin, an aptamer, an avimer, a camelid antibody, a designed ankyrin repeat protein (DARPin), a minibody, a nanobody, and a unibody.

13. An isolated polynucleotide encoding the isolated antibody, or antigen-binding fragment thereof, of claim 1.

14. An expression vector comprising the isolated polynucleotide of claim 13.

15. 15. An isolated host cell comprising the vector of claim 14.

16. A pharmaceutical composition comprising the isolated antibody, or antigen-binding fragment thereof, of claim 1 and a pharmaceutically acceptable carrier.

17. 17. The pharmaceutical composition of claim 16, wherein the composition has a purity of at least about 80%, 85%, 90%, 95%, 98%, or 99% on a protein basis with respect to the at least one antibody or antigen-binding fragment, and is substantially free of aggregates and endotoxins.

18. The composition optionally comprises V Ｌ 17. The pharmaceutical composition of claim 16, having reduced or undetectable heterogeneity of N-linked glycosylation in CDR3 sequences (optionally compared to the TS7 and 8E2 antibodies).

19. 17. The pharmaceutical composition of claim 16, wherein the composition is a sterile injectable solution suitable for intravenous, intramuscular, subcutaneous, or intraperitoneal administration, as the case may be.

20. 17. The pharmaceutical composition of claim 16 for the treatment of an IL-11-associated or IL-11-mediated disease or condition.

21. 21. The pharmaceutical composition of claim 20, wherein the disease or condition is cancer, an inflammatory disease, an autoimmune disease, a wasting disease, a bone disease, or a fibrotic disease.

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